NON-THERAPEUTIC METHOD FOR INCREASE THE GROWTH OF AN ANIMAL, NON-THERAPEUTIC USE OF ONE OR MORE COMP

专利摘要:
a method for increasing an animal's growth is provided. the method includes causing the animal to ingest or absorb an effective amount of one or more fe iii complex compounds, including, but not limited to, fe iii complexes comprising ligands attached to the iron core selected from amino acids or α-hydroxy acids, o-hydroxy benzoic acids or pyridine-2-carboxylic acids, such as ferric quinate, ferric tyrosine, ferric dopa and ferric phenylalanine. also disclosed are compounds that are structural and/or functional variants, derivatives and/or analogs of the foregoing compounds, as also described herein. methods for inhibiting, reducing, or preventing biofilm formation or accumulation on a surface; the treatment of, inhibition of growth of, and inhibition of colonization by, bacteria, both in biological and non-biological environments; disinfection of surfaces, potentiation of the effects of antibiotics and other antimicrobial agents, and increasing the sensitivity of bacteria and other microorganisms to antimicrobial agents are also provided.
公开号:BR112017002858B1
申请号:R112017002858-1
申请日:2015-08-11
公开日:2022-01-18
发明作者:Dlawer Ala'Aldeen；Jafar Mahdavi；Panos Soultanas
申请人:Akeso Biomedical, Inc；
IPC主号:

专利说明:

[1] This application claims the benefit and priority of USSN 62/188,183 filed July 2, 2015, USSN 62/171,081 filed June 4, 2015, USSN 62/138,499 filed March 26, 2015, USSN 62/137,630, filed March 24, 2015 and USSN 62/036,790, filed August 13, 2014, all of which are incorporated by reference in their entirety. FIELD OF THE INVENTION
[2] The present inventors have identified a class of compounds that have a wide variety of antimicrobial and other activities, particularly against bacteria, and have developed numerous uses and methods involving the compounds. In one aspect, the invention is generally directed to compositions, methods and uses for inhibiting, reducing or preventing the formation or accumulation of biofilm on a surface or for removing, dispersing, reducing or eradicating biofilm on a surface. In another embodiment, the invention also generally relates to compositions, methods and uses for treating, inhibiting the growth of, and inhibiting colonization by bacteria, in both biological and non-biological environments. In another embodiment, the invention also relates to compositions, methods, and uses for disinfecting surfaces, both in biological and non-biological environments, and products that have been coated with, or treated with, the compounds or compositions of the present invention. In another embodiment, the invention also relates to compositions, methods and uses for potentiating the effects of antibiotics and other antimicrobial agents, and increasing the sensitivity of bacteria and other microorganisms, including bacteria resistant to antibiotics, antibiotics and/or other agents. antimicrobial agents, and also to reverse antibiotic resistance in bacteria. In yet another embodiment, the invention also relates to compositions, methods and uses for increasing the growth of animals and their efficiency of use in food, in particular by oral administration of food and/or beverage compositions. FUNDAMENTALS OF THE INVENTION
[3] A biofilm is an accumulation of microorganisms (bacteria, fungi, and/or protozoa, with associated bacteriophages and other viruses) embedded in a polysaccharide matrix and adherent to biological or non-biotic solid surfaces. Biofilms are medically important, accounting for over 80% of hospital-acquired microbial infections in the body. Examples include infections of: oral soft tissues, teeth and dental implants; middle ear; gastrointestinal tract; urogenital tract; airway/lung tissue; eye; urinary tract prostheses; peritoneal membrane and peritoneal dialysis catheters, indwelling catheters for hemodialysis and for chronic administration of chemotherapeutic agents (Hickman catheters); cardiac implants, such as pacemakers, prosthetic heart valves, ventricular assist devices, and synthetic vascular grafts and stents; prostheses, internal fixation devices, percutaneous sutures; and tracheal and ventilatory tubes. Microorganisms tend to be much more resistant to antimicrobial agents and particularly difficult for the host's immune system to process an adequate response. Several bacterial pathogens have been shown to be associated with, and in some cases grow in biofilms, including Legionella pneumophila, S. aureus, Listeria monocytogenes, Campylobacter spp., E. coli O157:H7, Salmonella typhimurium, Pseudomonas, Vibrio cholerae, S. epidermidis, E. faecalis, and Helicobacter pylori.
[4] Biofilms are very difficult to treat with antimicrobials. Antimicrobial agents can be easily inactivated or fail to penetrate the biofilm. Furthermore, bacteria within biofilms have increased (up to 1000-fold) resistance to antimicrobial compounds, even though these same bacteria are sensitive to these agents if grown under planktonic conditions.
[5] Furthermore, bacteria embedded within biofilms are resistant to both the body's immunological and non-specific defense mechanisms. Contact with a solid surface triggers the expression of a panel of bacterial enzymes, which catalyze the formation of sticky polysaccharides that promote colonization and protection. The structure of biofilms is such that immune responses can be directed only to antigens found on the outer surface of the biofilm, and antibodies and other serum or salivary proteins are often unable to penetrate the biofilm. Furthermore, phagocytes are unable to effectively engulf a bacterium that grows within a complex polysaccharide matrix bound to a solid surface. This causes phagocytes to release large amounts of pro-inflammatory enzymes and cytokines, leading to inflammation and destruction of nearby tissues. Conventional therapy is characteristically ineffective against biofilms, as the minimum inhibitory concentration (MIC) of antimicrobial agents has been 10 to 1000 times higher than for planktonic organisms (Hoiby, et al., Int J Antimicrob Agents, 35 (Hoiby, et al., Int J Antimicrobial Agents, 35). 4): 322-32 (2010).
[6] It is an object of the present invention to provide compositions and methods for inhibiting or preventing the formation of biofilms or promoting the dissolution of biofilms from surfaces of interest.
[7] It is a further object of the present invention to provide methods for reducing the transmission of pathogens in the biofilm.
[8] It is yet another object of the present invention to provide methods for treating antibiotic resistant bacteria.
[9] It is yet another object to provide compositions to improve growth performance. SUMMARY OF THE INVENTION
[10] The present inventors have identified a class of compounds as described further in Section III. This application, which surprisingly has been shown to provide a wide range of activity, particularly against a variety of bacteria. The present invention provides numerous uses and methods involving the compounds, particularly in forming compositions. The present invention also provides compositions, articles and products which comprise one or more of the compounds as described below. The present invention also provides products produced by applying the numerous uses and methods of the present invention, as well as downstream products produced therefrom.
[11] In one embodiment, the present invention provides compounds as described further in Section III. This application and compositions comprising one or more of the compounds and methods and uses that employ one or more of the compounds and/or compositions to inhibit, reduce or prevent the formation or accumulation of biofilms on a surface or to remove, disperse, reduce or eradicate the biofilm on a surface. Accordingly, compositions are provided for inhibiting, reducing or removing biofilm accumulation in a subject and/or an article or other item. Exemplary compounds and compositions include an effective amount of one or more compounds selected from Ferric Quinate (Fe-QA, also referred to herein as FeQ) and ferric complexes with L-Tyrosine (Fe-Tyr), L-DOPA (Fe-DOPA ), L-phenylalanine (Fe-Phe) and hydrates, salts or derivatives thereof. See formulas IX, VIII and VII as defined below, respectively.
[12] As compositions they are effective against biofilms produced by a wide variety of microbial species including, without limitation, S. epidermidis, E. faecalis, E. coli, S. aureus, Campylobacter spp., H. pylori and Pseudomonas, isolated, or in combination.
[13] In one embodiment, an article or product, including medical devices having on or dispersed therein one or more of the compounds as described further in Section III. A of this application, or composition comprising one or more compounds, for example, of Formula IX (Fe-QA, or also referred to as FeQ), Formula VII (Fe-DOPA also referred to as FeDOPA) and Formula VIII (Fe-Tyr) or Fe -Phe, are designed to prevent or reduce the formation of a biofilm on the article or product, so as to prevent or reduce the formation of a biofilm on the medical device after implantation. The surface may be a biological surface (such as a surface of a living human, animal or plant surface, or the surface of a dead or harvested animal or plant), or a non-biological surface including, for example, plastics, polymers, biomaterials and metals. The present invention also provides products treated in accordance with this embodiment.
[14] In another embodiment, the invention provides compounds as described further in Section III. This application and compositions comprising one or more of the compounds, and methods and uses employing one or more of the compounds and/or compositions, for the treatment of, inhibiting growth and inhibiting colonization by bacteria, both in biological and non-biological environments .
[15] In another embodiment, the invention also relates to compounds and compositions comprising one or more of the compounds, and methods and uses that employ one or more of the compounds and/or compositions, for disinfecting surfaces, both in biological and non-biological environments. biologicals, and products that have been coated or treated by one or more of the compounds and/or compositions of the present invention.
[16] In another embodiment, the invention also relates to compounds and compositions comprising one or more of the compounds, and methods and uses that employ one or more of the compounds and/or compositions, to potentiate the effects of one or more antibiotics, increasing the sensitivity of bacteria (including antibiotic-resistant bacteria) to one or more antibiotics, and also to reverse antibiotic resistance in bacteria.
[17] In yet another embodiment the invention also relates to compounds and compositions comprising one or more of the compounds, and to methods and uses which employ one or more of the compounds and/or compositions, to increase animal growth and efficiency. for use in food, in particular by oral administration of food and beverage compositions.
[18] Methods of treating microbial infections in a subject by inhibiting, reducing or removing biofilm buildup in the subject and methods of treating subjects with microbial infections that are resistant to antibiotics are also provided. One method includes administering to the subject an effective amount of one or more compounds as described further in Section III. This application, including, but not limited to compounds according to Formula A or B as described herein, one or more compounds that bind to major outer membrane proteins (MOMPs) or Campylobacter FlaA, a histo group antigen -mimetic or synthetic human blood or a synthetic sugar. In one embodiment, the method includes administering to the subject an effective amount of a compound represented by Formula I, Formula II, Formula III, Formula IV, Formula V, and Formula VI. In a preferred embodiment, the method includes administering to the subject an effective amount of a compound represented by Formula IX (ferric quinate, Fe-QA also called FeQ), Formula VII (Fe-DOPA), Formula VIII (Fe-Tyr), or Fe-Phe, and hydrates, salts or derivatives thereof.
[19] In another preferred embodiment, the one or more compounds is as described further in Section III. A of this application, including, but not limited to, one or more compounds according to Formula A or B as described, one or more compounds of Formula I-IX or X-XIV, or compositions comprising one or more of said compounds , can be used to cure, treat or prevent symptoms of or associated with a variety of conditions as described herein, such as arterial lesions, gastritis, urinary tract infections, biliary tract infections, pyelonephritis, cystitis, sinus infections, ear infections , otitis media, otitis externa, leprosy, tuberculosis, conjunctivitis, blood infections, benign prostatic hyperplasia, chronic prostatitis, chronic lung infections of humans with cystic fibrosis, osteomyelitis, catheter infections, blood infections, skin infections, acne, rosacea, dental caries, periodontitis, gingivitis, nosocomial infections, arterial damage, endocarditis, periprosthetic joint infections, open or chronic wound infections, esophageal ulcers venous stasis, diabetic ulcers, arterial leg ulcers, pressure ulcers, endocarditis, pneumonia, orthopedic prosthesis and orthopedic implants, peritoneal dialysis peritonitis, cirrhosis and any other acute or chronic infection involving or associated with a biofilm.
[20] In a preferred embodiment for the treatment of antibiotic microbial infections (or other antimicrobial resistant microbial infections), the method includes administering to the subject an effective amount of an antibiotic or other antimicrobial agent (which may be the antibiotic/antimicrobial to which microbial infection is resistant) and an effective amount of one or more compounds as described further in Section III. A of this application, including, but not limited to, one or more compounds according to Formula A or B, as described herein, or one or more compounds represented by any of Formulas I to XIV, such as Formula I, Formula II, Formula III, Formula IV, Formula V and Formula VI or, preferably, an effective amount of one or more compounds represented by Formula IX, Formula VII and Formula VIII and hydrates, salts or derivatives thereof. The antibiotic/antimicrobial agent may be administered before one or more compounds according to the present invention, however, in a particularly preferred embodiment, the antibiotic/antimicrobial agent is administered simultaneously (as formulated in the same composition, or administered simultaneously, separate compositions) or after administration of the compounds of the present invention as described further in Section III. A of this patent application, including but not limited to compounds represented by Formula I, Formula II, Formula III, Formula IV, Formula V, Formula VI, Formula VII, Formula VIII, Formula IX and, Formula X, Formula XI, Formula XII, Formula XIII and Formula XIV. The antibiotic/antimicrobial agent and/or one or more compounds of the invention may also be incorporated into a medical device for administration.
[21] Methods for inhibiting the buildup of biofilm on a surface or for reducing or removing biofilm from a surface are also provided. The method includes contacting the surface with an effective amount of one or more as described later in Section III. A of this application, including, but not limited to, one or more compounds according to Formula A or B as described herein, compounds that bind to major outer membrane proteins (MOMPs) or FlaA from Campylobacter, an antigen of mimetic or synthetic human histo-blood or a synthetic serum, to inhibit or reduce biofilm accumulation or to reduce or remove surface biofilm. In a preferred embodiment, the method includes administering to the subject an effective amount of one or more compounds selected from Fe-QA, Fe-Tyr, Fe-DOPA and Fe-Phe, and hydrates, salts or derivatives thereof, to interfere with bacteria that bind to the surface or to each other. The surface to be treated can be contacted with the compounds by coating the surface with one or more compounds. In some embodiments, the surface is contacted by immersing the article to be treated in a composition comprising one or more compounds of the present invention, or washing, spraying, irrigating or cleaning the surface with a vehicle containing one or more compounds of the present invention. .
[22] The disclosed methods and uses are useful for inhibiting biofilm accumulation (or the reduction or removal of biofilm) produced by microbial species including S. epidermidis, E. faecalis, E. coli, S. aureus, Campylobacter spp. H. pylori and Pseudomonas, alone, or in combination, on/in a subject. The methods are thus also useful in treating diseases caused by these and other microorganisms that are associated with biofilm accumulation.
[23] With respect to surfaces, the disclosed methods and uses employing one or more compounds as described further in Section III. A of this application, including, but not limited to, one or more compounds according to Formula A or B as described herein, and more preferably one or more compounds selected from Fe-QA, Fe-Tyr, Fe-DOPA and Fe -Phe and hydrates, salts or derivatives thereof, are useful for inhibiting biofilm formation, dispersing biofilms and disinfecting articles, including, but not limited to, dental instruments, teeth, dentures, dental retainers, dental appliances, including braces plastic (such as Invisalign), medical instruments, medical devices, contact lenses and lens housings, catheters, surfaces (e.g. tables, countertops, bathtubs, tiles, filters, membranes, etc.), tubes, drains, pipes , including gas pipes, petroleum pipes, drill pipes, frac pipes, sewage pipes, drainage pipes, hoses, aquariums, showers, children's toys, boat hulls and cooling towers. Another embodiment of the present invention provides articles treated in accordance with the above methods and uses.
[24] In additional embodiments of the present invention, one or more compounds as described further in Section III. A of this application, including, but not limited to, one or more compounds according to Formula A or B as described herein, and more preferably one or more compounds selected from Fe-QA, Fe-Tyr, Fe-DOPA and Fe-Phe, and hydrates, salts or derivatives thereof, can be used in methods to make antifouling coatings, liquids, sprays and dispersants for bathroom and wound irrigation solutions.
[25] In additional embodiments of the present invention, one or more compounds as described further in Section III. A of this application, including, but not limited to, one or more compounds according to Formula A or B as described herein, and more preferably one or more compounds selected from Fe-QA, Fe-Tyr, Fe-DOPA and Fe-Phe and hydrates, salts or derivatives thereof, can also be used in cosmetic formulations, including skin treatments, acne treatments, toothpastes and mouthwash formulations. The compounds described herein can also be applied to toothbrush bristles, dental floss, and the like for oral health care.
[26] Compositions comprising one or more compounds as described further in Section III are also disclosed. A of this application, including, but not limited to, one or more compounds according to Formula A or B as described herein, and more preferably one or more compounds selected from Fe-QA, Fe-Tyr, Fe-DOPA and Fe-Phe and hydrates, salts or derivatives thereof, and methods using such compositions, to improve the growth performance of animals such as (including poultry, cattle, sheep, swine and goats) and other animals, as discussed later in section II. A, and as food supplements and formulas for these animals, in place of or in combination with existing bacteriostatic or bactericidal bacteria or growth enhancing compounds. In a preferred embodiment, the compositions can be administered to animals, such as cattle, to enhance growth performance. The compositions can also be used to decrease mortality-adjusted feed conversion ratios (MFCR). In a preferred embodiment, the method includes administering to the subject an effective amount of a compound represented by Formula IX, Formula VII and Formula VIII, or a hydrate, salt or derivative thereof. In a particularly preferred embodiment the compositions may be administered to chicken and other animals to promote growth. Additional related disclosure is provided in Section II. The one of this request, below. BRIEF DESCRIPTION OF THE FIGURES
[27] Fig. 1A is a bar graph showing biofilm formation by Enterococcus faecalis at time T = 0 in the presence of absence of different concentrations of Fe-QA. Fig. 1B is a bar graph showing biofilm formation by Enterococcus faecalis at time T = 24h in the presence of absence of different concentrations of Fe-QA.
[28] Fig. 2A is a bar graph showing biofilm formation by Staphylococcus epidermidis at time T = 0, in the presence of absence of different concentrations of Fe-QA. Fig. 2B is a bar graph showing biofilm formation by Staphylococcus epidermidis at time T = 24h in the presence of absence of different concentrations of Fe-QA.
[29] Fig. 3A is a bar graph showing biofilm formation by Staphylococcus aureus epidermidis at time T = 0, in the absence of different concentrations of Fe-QA. Fig. 3B is a bar graph showing biofilm formation by Staphylococcus aureus epidermidis at time T = 24h, in the presence of absence of different concentrations of Fe-QA.
[30] Fig. 4A is a bar graph showing binding of C. jejuni to BgAgs (common ABO blood histogroup antigens), Leb, and H-II, after culturing the bacteria in a medium that has Fe -QA at a concentration of 0.34 mM or 3.4 mM. Binding is shown after one passage and four passages (4 generations) with Fe-QA included in the medium, and compared to a control without Fe-QA. Fig. 4B is a bar graph showing binding of C. jejuni 11168 to BgAgs (ABO, Core-I, Core-II, HI, H-II, Leb, Ley, blood histogroup antigens). and Lex) and inhibition of this binding by Fe-QA (inhibitor)
[31] Fig. 5A is a bar graph showing the effect of Fe-QA treatment on Helicobacter pylori binding to human gastric tissue. Fig. 5B is a linear graph showing competitive inhibition of Leb binding to H. pylori by Fe-QA as the concentration of Fe-QA is increased. The plot is a plot of the ratio of Leb bound to free versus the concentration of Fe-QA.
[32] Fig. 6A is a bar graph showing the coverage rate of PAO1 Pseudomonas aeruginosa on the surface of a glass slide compared to the Pseudomonas medium only as a control, PAO1 Pseudomonas + 100 μM Fe treatment -QA, and Pseudomonas PAO1 without Fe-QA (X =Fe-QA). The graph shows that 100 μM Fe-QA ("X") inhibits biofilm formation by P. aeruginosa. Fig. 6B is a bar graph showing that Fe-QA inhibits biofilm formation by Uropathogenic E. coli (UPEC). The bar graph shows the coverage rate of UPEC on the surface of a glass slide compared to a UPEC-only control medium, and UPEC being cultured in the presence of 0.1 μM, 1 μM, 10 μM, and 100 μM concentrations of Fe-GQ.
[33] Fig. 7A is a graph showing the growth rate of UPEC in the presence of 100 μM Fe-QA and without Fe-QA over a 24 hour period. RPMI-1640 (bottom line); RPMI-1640 + UPEC (middle row); RPMI-1640 +100 μM FeQ+UPEC (top row). Fig 7B is a graph showing the growth rate of Pseudomonas aeruginosa in the presence of 100 μM Fe-QA and without Fe-QA. RPMI-1640 (bottom line); RPMI -1640 + UPEC (middle line); RPMI 1640 +100 μM FeQ+ PAO1 (upper row).
[34] Fig. 8A is a bar graph showing planktonic growth rates of wild-type and same-strain Campylobacter jejuni after mutation of T268 from MOMP. T268 from MOMP is replaced by glycine to form the MOMP-T strain. Fig. 8B is a bar graph showing the abilities of wild-type Campylobacter jejuni and the MOMP-T mutant to form biofilms.
[35] Fig. 9 is a graph showing the impact of Fe-QA on the growth rate of a kanamycin-resistant strain of E. coli. The groups, numbered from the top are as follows: (1) ΔsdiA-FeQ; (2) ΔsdiA-FeQ+Kan; (3) ΔsdiA-FeQ-Kan; (4) FeQ medium; (5) half-FeQ+Kan; (6) half-Kan. The graph shows the growth rate of the strain in the presence of Fe-QA (1) - top row), kanamycin ((2) - triangles) and a combination of Fe-QA and kanamycin ((3), squares). Three baselines are shown only for the medium containing Fe-QA, Fe-QA and kanamycin, and kanamycin alone.
[36] Fig. 10A shows the impact on the growth curve of the antibiotic resistant Enteropathogenic strain E. coli (EPEC) strain E2348/69 (EPEC wild-type O17:H6 genotype) when grown in the presence of (i) gentamicin ( 1.25 μM) (gray circles), (ii) Fe-Tyr (100 μM) (inverted gray triangle V), (iii) gentamicin (1.25 μM) and Fe-Tyr (1.25 μM) (white triangle vertical,Δ), and (iv) a control without gentamicin or Fe-Tyr present (black circles). Fig. 10B shows the growth rate of the Enteropathogenic E. coli (EPEC) strain E2348/69 when cultivated in the presence of (i), gentamicin (1.25 μM), (ii) Fe-Tyr (100 μM), ( iii) gentamicin (1.25 μM) and Fe-Tyr (1.25 μM), and (iv) a control without gentamicin or Fe-Tyr present.
[37] Figs. 11A-C show the impact on the growth curve of the antibiotic-resistant Enteropathogenic E. coli (EPEC) strain E2348/69 (EPEC wild-type O17:H6 genotype) when grown in the presence of gentamicin (1.25 μM) and increasing concentrations (10-68 μM) of Fe-QA (also known as FeQ). Fig. 11D compares the growth rates of the Enteropathogenic E. coli (EPEC) strain E2348/69, when cultivated in the presence of 10, 34, 68 and 100 μM Fe-QA. Fig. 11E compares the growth rates of the Enteropathogenic E. coli (EPEC) strain E2348/69 when grown in the presence of a fixed concentration of gentamicin (1.25 μM) and increasing concentrations of Fe-QA ranging from 10 to 100 μM .
[38] Fig. 12 quantitatively shows the difference in binding of EPEC cells to the surface of plastic well in the absence and presence of FeQ by measuring the optical absorbance of crystal violet which was absorbed by EPEC cells bound to the surface.
[39] Fig. 13 is a graph showing the impact of growth rates of an antibiotic-resistant clinical isolate of Pseudomonas in the presence of kanamycin, FeQ, and kanamycin plus FeQ compared to the strain cultured in the absence of kanamycin and FeQ .
[40] Figs. 14A-C show chemical structures of how FeQ can be conjugated to an agent that contains a reactive functional group suitable for immobilizing FeQ, for example, on a surface. Fig. 14A shows the conjugation of FeQ to a calix [4] arene structure that contains a photoreactive functional group. Fig. 14B shows the conjugation of FeQ to a calix [4] arene structure where the photoreactive functional group is positioned at a different location in the calix [4] arene structure compared to the structure of Fig. 14B. Fig. 14C shows the conjugation of FeQ to a calix[4] arene structure functionalized with two thiol groups.
[41] Figs. 15A and B are chemical structures illustrating how FeQ can be conjugated via a ligand to a substance that binds to a surface. In both structures, the ligand is spaced between the functional groups Y' and X', linked to FeQ via Y' and to hydroxyapatite (HA) via X'. The figures differ in the point of attachment to the quinic acid ligand.
[42] Fig. 16 is a graph showing that the wild-type O-glycosylated strain of Campylobacter dominates the infection of chickens colonized by a mixed population of O-glycosylated and non-glycosylated Campylobacter (MOMPT268G), and that the bacteria non-glycosylated are unable to colonize in a mixed population.
[43] Fig. 17 is a graph showing the average body weight (ABW) of broilers after 42 days of growth. The graph compares ABW to 42 days of chicken challenged with Campylobacter contaminated dirty sand on day 20 and treated from days 0-42 with FeQ or FeTyr for (i) a standard commercial target (2.979 kg) marked "Target" , (ii) a negative control (3.437 kg) marked "CNC" where the chicken was not challenged with Campylobacter contaminated dirty sand and (iii) a positive control (3.186 kg) marked "CC" where the chicken was challenged with Campylobacter - contaminated dirty sand. The graph shows that birds challenged with Campylobacter infected dirty sand had higher ABW at 42 days compared to the positive control (marked "CC") when treated with (iv) FeQ at 0.22 g/L in drinking water and FeQ at 0.22 g/Kg in food, marked "FeQ (W + F)" with an ABW of 3.342 kg, (v) FeQ at 0.22 g/L in drinking water, marked "FeQ (W)" with an ABW of 3.407 kg, (vi) FeQ at 0.22 g/kg in the feed, marked "FeQ (F)" with an ABW of 3.464 kg, (vii) FeQ at 0.022 g/drinking water, marked "FeQ (W )" with an ABW of 3.304 kg, and (viii) FeTyr at 0.02 g/L in drinking water, marked FeTyr (W) with an ABW of 3.341 kg.
[44] Fig. 18 is a graph showing the mortality-adjusted feed conversion ratio (MFCR) of broilers after 42 days of growth. The graph compares the MFCR at 42 days of chicken challenged with Campylobacter contaminated dirty sand on day 20 and treated from days 0-42 with FeQ or FeTyr for (i) a standard commercial target (1,703 kg) marked "Target" , (ii) a negative control (1.563 kg) marked "CNC" where the chicken was not challenged with Campylobacter contaminated dirty sand and (iii) a positive control (1.679 kg) marked "CC" where the chicken was challenged with Campylobacter - contaminated dirty sand. The graph shows that birds challenged with Campylobacter contaminated dirty sand had MFCR less than 42 days compared to the positive control (marked "CC") when treated with (iv) FeQ at 0.22 g/L in drinking water and FeQ at 0.22 g/Kg in food, labeled "FeQ (W + F)" with an MFCR of 1.595, (v) FeQ at 0.22 g/L in drinking water, labeled "FeQ (W)" with a MFCR of 1.560, (vi) FeQ at 0.22 g/kg in feed, labeled "FeQ (F)" with an MFCR of 1.563, (vii) FeQ at 0.022 g/L, labeled "FeQ (W)" with an MFCR of 1.612, and (viii) FeTyr at 0.02 g/L in drinking water, labeled FeTyr (W) with an MFCR of 1.577.
[45] Fig. 19 is a graph showing the number of Campylobacter colony forming units per gram (cfu/g) of bird droppings on day 42. The graph compares the day 42 cfu/g of chicken that were challenged with Campylobacter contaminated dirty sand on day 20 and treated from days 0-42 with FeQ or FeTyr for (i) a "CNC" labeled negative control (with a cfu/g of 28,000) where the chicken did not was challenged with Campylobacter contaminated litter, and (ii) a positive control labeled "CC" (with cfu/g of 1,280,000) where chickens were challenged with Campylobacter contaminated litter on day 21. The graph shows that the birds treated with FeQ or FeTyr had lower levels of Campylobacter in their droppings on day 42 when treated with (iii) FeQ at 0.22 g/L in drinking water and FeQ at 0.22 g/kg in feed, labeled "FeQ ( W + F)" with a cfu/g of 4.860, (iv) FeQ at 0.22 g/kg in the feed, labeled "FeQ (F)" with a cfu/g of 12.800, (v) FeQ at 0.022g/L in drinking water, labeled "FeQ(W)" with cfu/g of 900,000 and (vi) FeTyr at 0.02 g/L in drinking water, labeled FeTyr (W) with cfu/g of 16,600.
[46] Fig. 20 is a graph showing the average number of Campylobacter colony forming units per gram (cfu/g) of caeca samples on day 42. The graph compares the cfu/g on day 42 of chicken that were challenged with Campylobacter contaminated sand on day 20 and treated from days 0-42 with FeQ or FeTyr for (i) a negative control labeled "Treatment-1" where the chicken was not challenged with Campylobacter contaminated dirty sand , and (ii) a positive control labeled "Treatment-2" where the chickens were challenged with Campylobacter contaminated litter on day 21. The graph shows that birds treated with FeQ or FeTyr have lower levels of Campylobacter in their cage at day 42 when treated with (iii) FeQ at 0.22 g/L in drinking water and FeQ at 0.22 g/kg in feed, labeled "Treatment-3", (iv) FeQ at 0.22 g/L in water, marked "Treatment-5", (v) FeQ at 0.22 g/kg in food, marked "Treatment-6", (vi) FeQ at 0.022 g/L in drinking water, marked "Treat ento-7", and (vii) FeTyr at 0.02 g/L in drinking water, labeled "Treatment-8".
[47] Figs. 21A-C show the impact on the growth curve of the antibiotic-resistant Enteropathogenic E. coli (EPEC) strain E2348/69 (EPEC wild-type O17:H6 genotype) when cultivated in the presence of gentamicin (1.25 μM) and increasing concentrations (130-200 μM) of Fe-DOPA
[48] Fig. 22 quantitatively shows the difference in binding of EPEC cells to the plastic well surface in the absence and presence of FeDOPA (also referred to as Fe-DOPA) by measuring the optical absorbance of crystal violet that was absorbed by cells surface-bound EPEC.
[49] Fig. 23 shows 3 bar graphs at 24, 48 and 72 hours of the optical absorbance of crystal violet that was absorbed by the EPEC cells that remained attached to the surface of the plastic well after a mature biofilm formed by EPEC-pgA++ treated with FeTyr (shown as "FeY" in Fig. 23) at 100 μM, 150 μM and 200 μM compared to an untreated biofilm (marked "Control") in a crystal violet.
[50] Fig. 24A shows the analysis data in a positive way, as a graph of OPLS-DA scores. This shows a clear separation between fresh media (FM) and other used media (SMWT, SMWTF, SMMT, SMMTF). Fig. 24B also shows the analysis data in a positive way, where the fresh media (FM) results have been removed from the graph. Fig. 24C contrasts with Fig. 24A in that it shows the analysis data in a negative way. Fig. 24D contrasts with Fig. 24B in that it shows the analysis data in a negative way.
[51] Figs. 25A and B show the results of Example 30, which investigate the effects on antibiotic resistance of a laboratory strain of Psuedomonas aeruginosa (PAO1N) and a mixed population of clinical isolates (PAO Mixed), when incubated in Luria-Bertani (LB) medium ) isolated, or with different concentrations (34 μM, 100 μM, 200 μM and 340 μM) of FeQ or FePhe. Fig. 25A shows the results obtained with PAO1N cultures. Fig. 25B shows results with mixed cultures of PAO. DETAILED DESCRIPTION OF THE INVENTION I. DEFINITIONS
[52] "Aerosol", as used herein, refers to any preparation of a fine mist of particles, which may be in solution or a suspension, whether or not produced using a propellant.
[53] The term "alkyl" refers to the radical of saturated aliphatic groups, (i.e., an alkane with one hydrogen atom removed), including straight-chain alkyl groups, branched-chain alkyl groups, cycloalkyl (alicyclic) groups, alkyl-substituted cycloalkyl groups, and cycloalkyl-substituted alkyl groups.
[54] In preferred embodiments, a straight-chain or branched alkyl has 30 or fewer carbon atoms in its backbone (e.g., C1-C30 for straight chains, and C3-C30 for branched chains), preferably 20 or less, more preferably 15 or less, most preferably 10 or less. Likewise, preferred cycloalkyls may have 3 to 10 carbon atoms in their ring structure, and more preferably, have 5, 6 or 7 carbons in their ring structure. The term "alkyl" (or "lower alkyl") as used throughout the specification, examples and claims is intended to include both "unsubstituted alkyls" and "substituted alkyls", the latter referring to portions of alkyl with one or more substituents replacing a hydrogen on one or more carbons of the hydrocarbon backbone. Such substituents include, but are not limited to, halogen, hydroxyl, carbonyl (such as a carboxyl, alkoxycarbonyl, formyl, or an acyl group), thiocarbonyl (such as a thioester, a thioacetate, or a thioformate), alkoxyl, phosphoryl, phosphate, phosphonate, one-amino, amido, amidine, imine, cyano, nitro, azido, sulfhydryl, alkylthio, sulfate, sulfonate, sulfamoyl, sulfonamido, sulfonyl, heterocyclyl, aralkyl, or an aromatic or heteroaromatic moiety.
[55] Unless the number of carbons is specified otherwise, "lower alkyl" as used herein refers to an alkyl group, as defined above, but having from one to ten carbons, alternatively from one to about six carbon atoms in its main chain. Likewise, "lower alkenyl" and "lower alkynyl" have similar chain lengths. Throughout the application, preferred alkyl groups are lower alkyls. In preferred embodiments, a substituent designated herein as alkyl is lower alkyl.
[56] It will be understood by those skilled in the art that substituted moieties in the hydrocarbon chain may be substituted, if necessary. For example, substituents on a substituted alkyl may include halogen, hydroxy, nitro, thiols, amino, azide, imino, amido, phosphoryl (including phosphonate and phosphinate), sulfonyl (including sulfate, sulfonamido, sulfamoyl and sulfonate) and silyl groups, as well as ethers, alkylthio, carbonyls (including ketones, aldehydes, carboxylates and esters), -CF3 , -CN and the like. Cycloalkyls can be replaced in the same way.
[57] The term "heteroalkyl", as used herein, refers to carbon-containing, straight- or branched-chain, or cyclic radicals, or combinations thereof, containing at least one heteroatom. Suitable heteroatoms include, but are not limited to, O, N, Si, P, Se, B and S, wherein the phosphorus and sulfur atoms are optionally oxidized, and the nitrogen heteroatom is optionally quaternized. Heteroalkyls may be substituted as defined above for alkyl groups.
[58] The terms "alkenyl" and "alkynyl" refer to the unsaturated aliphatic groups analogous in length and possible substitution to the alkys described above, but which contain at least one double or triple bond, respectively.
[59] The terms "alkoxy" or "alkoxy" as used herein refer to an alkyl group, as defined above, having an oxygen radical attached thereto. Representative alkoxyl groups include methoxy, ethoxy, propyloxy, tert-butoxy and the like. "Ether" represents two hydrocarbons covalently linked by an oxygen. Thus, the substituent of an alkyl which makes that alkyl an ether is or resembles an alkoxyl, as may be represented by one of -O-alkyl, -O-alkenyl and -O-alkynyl. The terms "aroxy" and "aryloxy", as used interchangeably herein, may be represented by -O-aryl or O-heteroaryl, wherein aryl and heteroaryl are as defined below. The alkoxy and aroxy groups may be substituted as described above for alkyl.
[60] "Aryl", as used herein, refers to C5-C10 membered, aromatic, heterocyclic, fused aromatic, fused heterocyclic, biaromatic or biheterocyclic ring systems. Broadly, "aryl" as used herein includes 5-, 6-, 7-, 8-, 9- and 10-membered single ring aromatic groups which may include zero to four heteroatoms, for example benzene , pyrrole, furan, thiophene, imidazole, oxazole, thiazole, triazole, pyrazole, pyridine, pyrazine, pyridazine and pyrimidine, and the like. Those aryl groups with heteroatoms in the ring structure may also be referred to as "aryl heterocycles" or "heteroaromatics". The aromatic ring may be substituted at one or more ring positions with one or more substituents including, but not limited to, halogen, azide, alkyl, aralkyl, alkenyl, alkynyl, cycloalkyl, hydroxyl, alkoxyl, amino (or quaternized amino), nitro , sulfhydryl, imino, amido, phosphonate, phosphinate, carbonyl, carboxyl, silyl, ether, alkylthio, sulfonyl, sulfonamido, ketone, aldehyde, ester, heterocyclyl, aromatic or heteroaromatic moieties, -CF3 , -CN; and combinations thereof.
[61] The term "aryl" also includes polycyclic ring systems having two or more cyclic rings in which two or more carbons are common to two adjacent rings (i.e., "fused rings") in which at least one of the rings is aromatic. for example, the other cyclic ring or rings may be cycloalkyls, cycloalkenis, cycloalkynyls, aryls and/or heterocycles. Examples of heterocyclic ring include, but are not limited to, benzimidazolyl, benzofuranyl, benzothiofuranyl, benzothiophenyl, benzoxazolyl, benzoxazolinyl, benzothiazolyl, benzotriazolyl, benzotetrazolyl, benzisoxazolyl, benzisothiazolyl, benzimidazolinyl, carbazolyl, 4aH-carbazolyl, carbolinyl, chromanyl, chromenyl, decahydroxyquinolinyl, , 2H, 6H-1,5,2-dithiazinyl, dihydrofuro[2,3-b]tetrahydrofuranyl, furanyl, furazanyl, imidazolinyl, imidazolinyl, imidazole, 1H-indazolyl, indolenyl, indolinyl, indolizinyl, indolyl, 3,H -indolyl, isatinoyl, isobenzofuranyl, isochromanyl, isoindazolyl, isoindolinyl, isoindolyl, isoquinolinyl, isothiazolyl, isoxazolyl, methylenedioxyphenyl, morpholinyl, naphthyridinyl, octahydroisoquinolinyl, oxadiazolyl, 1,2,3-oxadiazolyl, 1,2,4-oxadiazolyl, 1, 2 ,5-oxadiazolyl, 1,3,4-oxadiazolyl, oxazolidinyl, oxazolyl, oxindolyl, pyrimidinyl, phenanthridinyl, phenanthrolinyl, phenazinyl, phenothiazinyl, phenoxatinyl, phenoxazinyl, phthalazinyl, piperazinyl, piperidinyl, piperi donyl, 4-piperidonyl, piperonyl, pteridinyl, purinyl, pyranyl, pyrazinyl, pyrazolidinyl, pyrazolinyl, pyrazolyl, pyridazinyl, pyridooxazole, pyridoimidazole, pyridothiazole, pyridinyl, pyridyl, pyrimidinyl, pyrrolidinyl, pyrrolinyl, 2H-pyrrolyl, pyrrolyl, quinazolinyl, quinolinyl, 4H-quinolizinyl, quinoxalinyl, quinuclidinyl, tetrahydrofuranyl, tetrahydroisoquinolinyl, tetrahydroquinolinyl, tetrazolyl, 6H-1,2,5-thiadiazinyl, 1,2,3-thiadiazolyl, 1,2,4-thiadiazolyl, 1, 2,5-thiadiazolyl, 1,3,4-thiadiazolyl, thianthrenyl, thiazoyl, thienyl, thienothiazolyl, thienooxazolyl, thienoimidazolyl, thiophenyl and xanthenyl. One or more of the rings may be substituted as defined above for "aryl".
[62] The term "aralkyl", as used herein, refers to an alkyl group substituted with an aryl group (e.g., an aromatic or heteroaromatic group).
[63] The term "aralkyloxy" may be represented by -O-aralkyl, where aralkyl is as defined above.
[64] "Biofilm" as used herein refers to any group of microorganisms in which cells adhere to one another on a surface.
[65] A "cream" is a viscous liquid or semi-solid emulsion, either of the "oil-in-water" or "water-in-oil" type.
[66] An "emulsion" is a composition containing a mixture of immiscible components homogeneously mixed together.
[67] "Gel" as used herein is a colloid in which the dispersed phase has combined with the continuous phase to produce a semi-solid material, such as gelatin.
[68] "Cleaning formulation", as used herein, means a composition suitable for application to a surface to remove dirt and oils, for disinfection, or a combination thereof. Cleaning formulations can be antibacterial, antimicrobial, or both. Cleansing formulations are suitable for use on human skin when none of the components of the composition are present in concentrations that cause significant signs of irritation when applied to human skin. As used herein, "significant signs of irritation" include erythema, redness and/or swelling at the injection site or at the application site, application site necrosis, exfoliative application site dermatitis, and severe pain that impedes daily activity and /or requires medical attention or hospitalization. Cleaning formulations may be suitable for use in the human oral cavity. Cleaning formulations may be suitable for use with articles which, after exposure and optionally with residual levels of cleaning composition present on and/or in the article, will then be contacted with human skin or another part of the human body, such as in that the article (eg a denture) will be brought into contact with the oral cavity or will be contacted with the eye (eg a contact lens). The cleaning formulations may be suitable for use with foodstuffs and/or their packaging and may, for example, be suitable for cleaning meat products and/or carcasses used in the production of meat products. Cleaning formulations may be suitable for equipment used in the production of cleaning foods. Cleaning formulations may be suitable for use in cleaning medical devices, including implantable medical devices. Many other types of cleaning formulations can also be provided by the present invention, other examples of which are discussed in further sections of this application.
[69] "Chronic wound" as used herein refers to a wound that fails to progress through an orderly and timely sequence of repair or a wound that does not respond to treatment and/or the demands of treatment are beyond physical health. of the patient, tolerance or resistance. Many wounds that are considered primarily acute wounds eventually turn into chronic wounds due to factors not yet understood. A significant factor is the transition of planktonic bacteria within the wound to form a biofilm.
[70] The term "heteroatom" used in this instrument refers to an atom of any element other than carbon or hydrogen. Examples of heteroatoms are boron, nitrogen, oxygen, phosphorus, sulfur and selenium. Other useful heteroatoms include silicon and arsenic.
[71] "Inhibition" or "inhibiting" biofilm formation, as used herein, refers to a decrease in the formation and/or growth of biofilm-associated microorganisms.
[72] A "lotion" is a low to medium viscosity liquid formulation.
[73] As used herein, the term "nitro" means -NO2; the term "halogen" means -F, -Cl, -Br or -I; the term "sulfhydryl" means -SH; the term "hydroxyl" means -OH; and the term "sulfonyl" means -SO 2 -.
[74] "Oil" as used herein refers to a composition containing at least 95% by weight of a lipophilic substance. Examples of lipophilic substances include, but are not limited to, synthetic and naturally occurring oils, fats, fatty acids, lecithins, triglycerides and combinations thereof.
[75] An "ointment" is a semi-solid preparation containing an ointment base and optionally one or more active substances.
[76] "Parenteral administration", as used herein, means administration by any method other than via the digestive tract or non-topical or regional non-invasive routes.
[77] "Patient" or "subject" to be treated and/or used in accordance with any aspect of the present invention as described herein refers to a human or non-human animal such as a primate, non-human primate, laboratory animal, farm animal, livestock, or a domestic pet. Examples of animals may optionally include chickens, particularly a meat-like chicken, such as a broiler, or a laying hen, such as a pullet or hen, or a broiler. Also optionally included, without limitation, are other poultry, such as a turkey, geese, quail or ducks, or livestock, such as cattle, sheep, goats or swine, alpaca, banteng, bison, camel, cat, deer, dog , donkey, gayal, e.g. zoo animals, game animals, game animals fish (including freshwater and saltwater fish, farmed fish and ornamental fish), other marine and aquatic animals including crustaceans such as, e.g. oysters, mussels, clams, shrimp, lobsters, crayfish, crabs, cuttlefish, octopus and squid, domestic animals such as cats and dogs, rodents (such as mice, rats, guinea pigs, hamsters) and horses, as well as any other domestic, wild and farmed animals, including mammals, marine animals, amphibians, birds, reptiles, insects and other invertebrates.
[78] "Pharmaceutically acceptable", as used herein, refers to compounds, materials, compositions and/or dosage forms that are, within the scope of medical judgment, suitable for use in contact with the tissues of humans and animals ( such as one or more of animal "patients" or "subjects" as discussed above) without excessive toxicity, irritation, allergic response, or other problems or complications commensurate with a reasonable benefit/risk ratio.
[79] "Pharmaceutically acceptable salt", as used herein, refers to derivatives of the compounds defined herein, wherein the parent compound is modified by producing its acidic or basic salts.
[80] "Therapeutically effective" or "effective amount" as used herein means that the amount of the composition used is an amount sufficient to ameliorate one or more causes or symptoms of a condition, bacterial colonization, disease or disorder. Such an improvement only requires a reduction or alteration, not necessarily elimination. As used herein, the terms "therapeutically effective amount" "therapeutic amount" and "pharmaceutically effective amount" are synonymous. One skilled in the art can readily determine the appropriate therapeutic amount.
[81] The term "substituted", as used herein, refers to all permissible substituents of the compounds. In a broad aspect, permissible substituents include acyclic and cyclic, branched and unbranched, carbocyclic and heterocyclic, aromatic and non-aromatic substituents of organic compounds. Illustrative substituents include, but are not limited to, halogens, hydroxyl groups or any other organic groups containing any number of carbon atoms, preferably 1-14 carbon atoms, and optionally include one or more heteroatoms such as oxygen, sulfur or nitrogen grouping in linear, branched or cyclic structural shapes. Representative substituents include alkyl, substituted alkyl, alkenyl, substituted alkenyl, alkynyl, substituted alkynyl, phenyl, substituted phenyl, aryl, substituted aryl, heteroaryl, substituted heteroaryl, halo, hydroxyl, alkoxy, substituted alkoxy, phenoxy, substituted phenoxy, aroxy, substituted aroxy, alkylthio, substituted alkylthio, phenylthio, substituted phenylthio, arylthio, substituted arylthio, cyano, isocyano, substituted isocyano, carbonyl, substituted carbonyl, carboxyl, substituted carboxyl, amino, substituted amino, amido, substituted amido, substituted sulfonyl sulfonyl, sulfonic acid, phosphoryl , substituted phosphoryl, methylphosphonyl, substituted methylphosphonyl, polyaryl, substituted polyaryl, C3-C20 cyclic, substituted C3-C20 cyclic, heterocyclic groups, substituted heterocyclic, amino acids, peptides and polypeptide.
[82] "Treatment", "treating", or "alleviating", as used in connection with a disease or infection, refers to an intervention performed with the intention of altering or inhibiting the pathology of a disorder. II. ASPECTS OF THE INVENTION
[83] While aspects of the invention are described throughout the application, some of the main aspects, which all utilize the compounds of the present invention as described further in section III.A of this application, can be summarized as:
[84] (i) Increased animal growth;
[85] (ii) Enhancing the effect of antibiotics and other antimicrobial agents and addressing antibiotic resistance;
[86] (iii) Inhibition of the formation and treatment of preformed biofilms; treatment of microbial infections that reduce microbial colonization; and disinfection of surfaces;
[87] (iv) Compounds of the present invention as described in section III. A of this application, and compositions comprising one or more of said compounds.
[88] These and other aspects of the present invention, and other embodiments of these aspects, will be discussed in more detail below. A. Enhancement of animal growth
[89] A first aspect of the present invention is based on the surprising finding that the compounds of the present invention as described further in section III.A of this application can be used to enhance the growth of animals. Numerous examples of this effect are provided in Example 18 of this application, as discussed further below.
[90] Accordingly, the first aspect of the present invention provides a method of increasing the growth of an animal, the method comprising causing the animal to ingest and/or absorb an effective amount of one or more compounds having the structure of Formula A or B, or other compounds of the invention as described further in section III.A of this application below.
[91] In other words, the first aspect of the present invention also provides the use of one or more compounds having the structure of Formula A or B, or other compounds of the invention, as described further in section III.A of this application below. , to enhance the growth of an animal by causing the animal to ingest an effective amount of one or more compounds.
[92] Typically, in the practice of the first aspect of the present invention, the one or more compounds will be presented directly to the animal for ingestion and/or absorption. However, in an alternative optional embodiment of the first aspect of the present invention, the animal may be caused to ingest or absorb one or more compounds having the structure of Formula A or B, or other compounds of the invention as described in section III.A hereof. application below, providing the animal simultaneously, separately or sequentially with components that cause the animal to form an effective amount of the one or more compounds having the structure of Formula A or B or other compounds of the invention as described further in section III.A hereof. order below, in situ. For example, the animal could be provided with a source of ferrous sulfate and simultaneously, separately or sequentially with a source of quinic acid or a salt thereof (or other α-hydroxy acid), or it could be provided with a source of ferrous sulfate and simultaneously, separately or sequentially with a source of a natural or synthetic amino acid, such as L-tyrosine, L-DOPA or L-phenylalanine.
[93] In a preferred option of the first aspect of the present invention, the animal ingests and/or absorbs one or more compounds having the structure of Formula A as described further in section III.A of this application below and in another preferred option, the one or more compounds are selected from the group consisting of a complex of an amino acid with Fe III and a complex of an α-hydroxy acid with Fe III, or salts and/or hydrates thereof. In particularly preferred options of the first aspect of the present invention, one or more compounds may or may not be selected from any one or more of the group consisting of a complex of quinic acid with Fe III (such as a complex with the structure of Formula IX) , a complex of L-tyrosine with Fe III (such as a complex having the structure of Formula VIII), a complex of L-DOPA with Fe III (such as a complex having the structure of Formula VII), and a complex of L- phenylalanine with Fe III. Accordingly, in one embodiment of the first aspect of the invention, a complex of L-tyrosine with Fe III (such as a complex having the structure of Formula VIII) is particularly preferred. Optionally, one or more compounds is not a complex of quinic acid with Fe III (such as a complex having the structure of Formula IX).
[94] The animal can be made to ingest or absorb one or more compounds having the structure of Formula A or B, or other compounds of the invention as described further in section III.A of this application below, providing one or more compounds (or its component parts to form the compound(s) in situ) by dietary means, such as in or mixed with an animal feed, as a dietary supplement, and/or in drinking water. Another option, in the case of marine, aquatic, amphibian or other animals that live partially or completely in water, is to add one or more compounds (or components thereof to form the compound(s) in situ), such as treatment of ponds that contain farmed fish or crustaceans, such as shrimp and crayfish. Therefore, for example, in a preferred embodiment, one or more compounds may be presented to the animal through one or more routes selected from the group consisting of an animal feed, an animal feed supplement, and in drinking water or by exposure to other waters. It should be noted that, depending on the solubility of the one or more compounds used, it may be beneficial to introduce a co-solvent to solubilize to aid dissolution in water at an effective concentration.
[95] Accordingly, in another embodiment of the first aspect of the present invention, there is provided an animal feed, an animal feed supplement and a supply of drinking water, each comprising one or more compounds having the structure of Formula A or B, or other compounds of the invention as described further in section III.A of this application below. Suitable concentrations of one or more compounds to be included in animal feed, animal feed supplement and a potable water supply include concentrations as discussed further below.
[96] Also provided herein, in another embodiment of the first aspect of the present invention, is a method for producing an animal feed product or an animal feed supplement product, the method comprising the steps of incorporating one or more compounds having the structure of Formula A or B or other compounds of the invention, as described further in section III.A of this application below, in the animal feed product or animal feed supplement product during the preparation of the food or supplement. One or more compounds may be incorporated into the product at any stage during the production process and may, for example, be included prior to one or more heating steps, such one or more heating steps comprising exposing a composition comprising a or more compounds at a temperature greater than 50°C, greater than 60°C, greater than 70°C, greater than 80°C, greater than 90°C or greater than 100°C, and preferably where exposure to temperature is in a selected range of 50-200 °C, 60-150 °C, 70-100 °C. In some embodiments, a temperature range for a heating step may be in the range of 70-90°C, such as 75-88°C, 80-87°C, 81-86°C, or 82-85°C .
[97] Optionally, in one embodiment, a suitable method for producing an animal feed, such as a feed for a chicken (including a broiler) may include the steps of: (a) combining nutritional and/or other components dietary components (such as one or more components selected from wheat, soybeans, soybean oil, minerals and other additives) to form a grain or other mixture; (b) heating the grain or other mixture in a heating step, as described above, such as with steam at 85°C for a time effective to kill any pathogens, such as Salmonella. A period of 5-10 minutes, such as 6-8 minutes, is an example of an effective period at 85°C, although the time can be adjusted depending on the temperature used; (c)cooling the heated mixture. Preferably, cooling is conducted at a rate and under conditions effective to prevent condensation from forming, as condensation can result in the growth of pathogens including Salmonella. (d) optionally, compressing the cooled mixture; (e) forming feed pellets from the cooled mixture, such as by pelleting using an extruder which heats the feed to a suitable temperature, as discussed above, for example in the range 82-85°C; (f) adding heat sensitive additives, typically by spraying. Heat sensitive additives may include enzymes, which may (for example) be selected from the group consisting of phytase, xylase, beta-lactamase.
[98] According to the above method for producing an animal food product, the method comprising the step of incorporating one or more compounds having the structure of Formula A or B, or other compounds of the invention as described later in the section III.A of this application below, in the animal food product at any one or more stages of production, including during step (a), between steps (a) and (b), during step (b), between steps (b) and (c), during step (c), between steps (c) and (d), during step (d), between steps (D) and (e), during step (e) , between steps (e) and (f), during step (f) or after step (f).
[99] Other additives that may be included at the time of addition of the heat sensitive additives or at earlier stages include one or more additives selected from the list consisting of creatine, amino acids (eg threonine) and salt.
[100] An animal feed or animal feed supplement as described herein and useful in the context of the first aspect of the present invention, or any other aspect of the present invention, may be a vegetarian or non-vegetarian product. A vegetarian product does not contain meat or fish products. A non-vegetarian diet may contain either a fish product (such as fishmeal) or a meat product (such as meat products, bone meal, etc.).
[101] Also provided herein, in another embodiment of the first aspect of the present invention, is a method for producing a drinking water for animals, the method comprising adding one or more compounds having the structure of Formula A or B, or other Compounds of the invention as described further in Section III.A of this application below, in a supply of drinking water for animals. Suitable concentrations of the one or more compounds in a potable water supply are as discussed below and are typically at a concentration effective to produce the growth-enhancing effect in accordance with the first aspect of the present invention. Determining an appropriate concentration can take into account the amount of drinking water consumed by the animal. For example, a UK broiler (or at a temperature equivalent to that used in the UK) typically consumes a daily amount of drinking water dependent on its age which can be calculated with reference to the chicken's age in days multiplied by approximately 4 -10ml, such as 5-9ml, 68ml, for example about 7.14ml. So, for example, a 42 day old broiler may have a daily water consumption of 168 mL to 420 mL per day, more typically around 300 mL per day ± 30%, 20%, 15%, 10%, 5 %, 4%, 3%, 2% or 1%. Broiler chickens raised at different temperatures may consume more (e.g. in the southern US where summer temperatures will be high and water consumption could be higher, especially in sheds where the temperature is not controlled), or less water .
[102] The animal may ingest or absorb an effective amount of one or more compounds on a regular and repeated basis. For example, the animal may ingest or absorb an effective amount of one or more compounds weekly, daily, daily, or more than once daily during performance of the method or use. In one option, one or more compounds are included in the animal feed, an animal feed supplement, and/or in drinking water, and the animal ingests one or more compounds when eating and/or drinking and, optionally, eating and/or drinking. This ingestion or absorption of an effective amount of one or more compounds may continue over a period of growth of the animal which may correspond to a period of time which is, is up to, or is at least 5%, 10%, 25%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 95% or substantially 100% of the animal's life from birth to death. Ingestion or absorption of an effective amount of one or more compounds may begin on the day of the animal's birth, or at the age of 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12 , 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39 , 40, 41, 42, 43, 44, 45, 46, 47 days or more. After the animal begins to ingest or absorb one or compounds, the animal may continue to do so on a regular and repeated basis over a period of time that may be, or be up to, or at least 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27 28, 29 , 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47 days or more.
[103]In the case of chickens, especially broilers, it may be preferable that the chickens ingest one or more compounds according to the present invention on a regular and repeated basis on a starter diet, on a producer diet and/or or on a finisher diet, as described later. Chickens raised for other purposes, such as broilers and/or laying hens, typically receive diets that are different from that of broilers, as discussed later in this application and standard diets for broilers and laying hens are well known to those of skill. in the technique. In accordance with other embodiments of the first aspect of the present invention, the one or more compounds having the structure of Formula A or B, or other compounds of the invention, as described further in section III.A of this application below, are incorporated into a food for animals for a breeding hen and/or a laying hen.
[104] In one embodiment of the first aspect of the present invention, the one or more compounds may be included in an animal feed, or in an animal feed supplement, for commercial poultry feeds such as chickens, turkeys, pheasants and ducks. . In one option, the one or more compounds may be included, or used to supplement, a poultry feed, which may be a "complete" feed. A complete feed is designed to contain all the protein, energy, vitamins, minerals and other nutrients needed for proper growth, egg production and bird health. Feeding any other ingredient, mixed with the feed or fed separately, in addition to using a whole feed, can upset the balance of nutrients in the "whole" feed. Adding additional grains or supplementation to the complete poultry feed is not recommended.
[105] Broilers used in commercial production of optimized broilers are typically fed different diets depending on their age. For example, broiler chickens can be raised using three diets. These diets are typically called "starter", "growth" and "finish". "Pre-starter" diets are also possible.
[106] "Starter", "Growth", and "Finish" are typically distinguished by the crude protein content, which is often provided by ingredients such as soy flour (SBM). For example, a broiler starter diet may optionally contain a crude protein content of about 22-25% by weight, such as 22%, 23%, 24% or 25%, with 23 or 25 % being preferred. In another example, a growth diet for a broiler may optionally contain a crude protein content of about 21-23% by weight, such as 21%, 22% or 23%, with 22% being preferred. In another example, a finisher diet for a broiler may optionally contain a crude protein content of about 19-23% by weight, such as 19%, 20%, 21%, 22% or 23%, with 19%, 20%, or 21% being preferred.
[107]Additionally or alternatively, "starter", "growth" and "finisher" can be distinguished by the metabolizable energy (ME) content, which is typically lowest for the starter diet and highest for the diet. with the grower diet having a level between the two. For example, a starter diet of a broiler may have an ME of 3000 or about 3025 kcal/kg (± 50, 40, 30, 20, 10, 5 or less kcal/kg.) In another example, a growth diet for broilers may have an ME of about 3100 or 3150 kcal/kg (± 50, 40, 30, 20, 10, 5 or less kcal/kg.) In another example, a growth diet for a broiler might have an ME of about 3200 kcal/kg (± 50, 40, 30, 20, 10, 5 or less kcal/kg) .
[108]Animal feeds, including chicken and, more particularly, feeds for broilers, according to the present invention may also typically contain one or more (preferably all) of the following:
[109] Macrominerals, which include those selected from the group consisting of calcium, phosphorus, magnesium, sodium, potassium, and chloride.
[110]Trace minerals, including zinc and/or selenium.
[111] Added vitamins, which include those selected from the group consisting of vitamin A, nicotinic acid, pantothenic acid, pyridoxine (B6) and biotin in corn and wheat-based foods. In addition, there is a basic requirement of broilers for vitamin E at 1015 mg/kg. The need for extra vitamin E supplementation will depend on the level and type of fat in the diet, the level of selenium, and the presence of pro and anti-oxidants. Heat treatment of foods can result in the destruction of up to 20% of vitamin E. Choline can also be given in a complete food.
[112] Non-nutritive food additives may also be included. Enzymes are routinely used in poultry feed to improve the digestibility of feed ingredients. In general, food enzymes are available that act on carbohydrates, minerals and proteins. Non-polysaccharide starch enzymes (NSP) are economically beneficial in wheat-based foods. These enzymes will also allow greater flexibility in the levels of barley to be included in the ration. Phytase enzymes can be used to improve phosphorus utilization from phytate. Protease enzymes can be included to act on plant products. Carbohydrase enzymes can be added and may provide beneficial responses when used in corn-soy diets. When adding enzymes prior to thermal processing of broiler feeds, there is the potential for a loss in enzyme activity. This can be avoided by spraying enzymes on the food at the end of processing.
[113] Medicinal and prophylactic drugs (other than compounds as defined in section III.A below) may be added. A wide variety of drugs and coccidiostats and antibiotics can be administered through food. Antibiotic Growth Promoters/Digestion Enhancers may be included and may, for example, provide a mode of action that involves modification of the gut microflora, with consequent benefits in nutrient utilization.
[114] Prebiotics can be added and refer to a group of substances that stimulate the growth of beneficial microorganisms at the expense of harmful microorganisms. Oligosaccharides form the largest group of these products at the moment.
[115] Probiotics can be added to introduce live microorganisms into the digestive tract to aid in the establishment of a stable and beneficial microflora. The objective is to provide the intestine with positive, non-pathogenic microorganisms that will prevent colonization with pathogenic microorganisms by competitive exclusion.
[116] Organic acids may be added. Organic acid products can be used to reduce bacterial contamination of food (eg after heat treatment) and can also encourage beneficial microflora to develop in the bird's digestive tract.
[117]Absorbents are used specifically to absorb mycotoxins. They can also have a beneficial effect on the bird's overall health and nutrient absorption. There are a variety of products available for use as absorbents, including various clays and charcoal.
[118] Antioxidants may provide important protection against nutrient loss in broiler feed. Some food ingredients, for example fishmeal and fats, can be protected. Vitamin premixes should be protected by an antioxidant unless optimal storage times and conditions are provided. Additional antioxidants may be added to the final feed where prolonged storage or inadequate storage conditions are unavoidable.
[119]Anti-mold agents can be added. For example, mold inhibitors can be added to feed ingredients, which have become contaminated, or to finished feeds to reduce mold growth and mycotoxin production.
[120]Pelleting agents can be added, and are used to improve pellet hardness. Some examples of pellet binders are hemicellulose, bentonite and guar gum.
[121] Other products of possible use in broiler production include essential oils, nucleotides, glucans and specialized plant extracts. In areas of the world where its use is permitted, formaldehyde can be used to treat/preserve food.
[122] Without limitation, exemplary "starter", "growth" and "finisher" diets include those shown in Example 18 of this application, below.
[123] The broiler starter diet can be fed for about 10-12 days (typically within the first 7-14 days of life). This starter diet can be followed by the growth diet, which is fed to broilers for almost 2 weeks (typically from the age of around 11-24 days, although in any case after the end of using the broiler diet). initiation). The finisher diet can be used for the remainder of the production period (typically from the age of around 24, or 25, days until harvest). Some broiler houses will use more or less diets (eg 4 diets), and will vary the timing of diet changes. Broilers are typically harvested between 35 and 42 days, although this time can be longer or shorter. The UK market normally harvests on the 30th-35th. Other countries, including some European countries, harvest after 25 days, although more typically after 30 days. However, other countries such as the US typically harvest within 42-47 days. Chickens that are not broilers, including free-range chickens, can be harvested at later ages. In the context of practicing the first aspect of the present invention, any age of harvest may be used, although more typically (e.g. in the context of broiler chickens) after the start of the finisher diet, and optionally (and without limitation) at any of the days 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49 , 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69 or 70, up to about 11 weeks, 12 weeks, 13 weeks, 14 weeks, 15 weeks, 16 weeks, 17 weeks, 18 weeks, 19 weeks, 20 weeks or more.
[124] In some embodiments of the first aspect of the present invention, methods for producing broilers or other animals in groups that are single-sex (i.e., groups of all-female or all-male animals) and/or may be performed in mixed male groups (i.e. mixed males and females). For example, in the case of broiler production, it may be appropriate to select and re-articulate a single sex group of male roosters, and it may be appropriate to harvest roosters earlier than female or mixed sex groups. For example, a single sex rooster can be harvested from broilers aged around 30 days or, in other options, aged any one or more than 25, 26, 27, 28, 29, 30, 31 , 32, 33, 34, 35 or more days. For example, at age 30 days, a group of untreated rooster may have an average target weight of about 1.95 kg, whereas in the case of increased growth resulting from performance of the method of the first aspect of the present invention, it may be appropriate to harvest roosters at an earlier stage with the defined target weight, or harvest at the same age and with a higher average weight, or at the same age and target weight with the use of reduced feed consumption due to higher feed conversion efficiency. In another example, a mixed sex group of broilers aged around 35 days or, in other options, aged any one or more than 25, 26, 27, 28, 29, 30, can be harvested. 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50 or more days. For example, at 35 days of age, an untreated mixed-sex group may have an average target weight of about 2.1-2.2 kg, whereas in the case of increased growth resulting from the performance of the first aspect of In the present invention, it may be appropriate to harvest the mixed sex group at an earlier stage with a defined target weight, or to harvest at the same age and a higher average weight, or at the same age and target weight with the use of reduced food consumption for animals due to higher feed conversion efficiency.
[125] In accordance with the practice of the first aspect of the invention, for the purpose of enhancing the growth of broilers, the one or more compounds may be included in any one, two or three of the starter, grower and breeder diets. finishing. In one embodiment, the one or more compounds may be included in the starter diet only. In another embodiment, the one or more compounds may be included in the growth diet alone. In another embodiment, the one or more compounds may be included only in the finishing diet. In another embodiment, the one or more compounds can be included only in the starter and grower diets, but not in the finisher diet. In another embodiment, the one or more compounds can be included only in the starter and finisher diets, but not in the growth diet. In another embodiment, the one or more compounds can be included only in the grow and finish diets, but not in the starter diet. In another embodiment, the one or more compounds may be included in all starter, grower and finisher diets.
[126] In accordance with other embodiments of the first aspect of the present invention, the animal to be cultured may be a laying hen. A typical process of raising a laying hen might involve starting egg production at around 23 weeks of age, and slaughtering at around 60 weeks of age. The laying hen may be treated in accordance with the first aspect of the present invention prior to the initiation of egg-laying, and/or during egg-laying and/or until the time of slaughter. Treatment may, for example, last for about 5, 10, 15, 20, 25, 30, 35, 40, 45, 50, 55 or 60 weeks; the term "about" in this context can include the meaning of ± 4, 3, 2 or 1 weeks of the indicated value. Considering that laying hens typically begin laying eggs at 23 weeks of age, taking advantage of the increased growth and/or improved feed utilization of the first aspect of the present invention, it may be appropriate to start egg production at a later stage. early age, such as at 18, 19, 20, 21 or 22 weeks of age. Furthermore, by taking advantage of the increased growth and/or improved feed utilization of the first aspect of the present invention, the present invention can be used to achieve an effect (compared to an untreated control group that is raised under identical conditions, except for the application of the method of the first aspect of the present invention) selected from: (a) production of better quality eggs. The improved quality can, for example, be selected from size, shell quality, air cell, white and yolk. Shell quality is determined from one or more sizes, visual defects, specific gravity, color, breaking strength, shell percentage (shell weight x 100/egg weight), shell thickness and egg ultrastructure. Improved quality may be reflected in a higher proportion of eggs classified as US grade A or AA (e.g. the US standard for grading eggs is discussed at http://www.fao.org/docrep/005/y4628e/y4628e04 .htm, the contents of which are incorporated herein by reference); (b) the production of eggs of increased size (e.g. with a weight of up to or at least 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%% , 10%, 15%, 20% or more); and/or (c) egg production in increased numbers (such as an average daily amount, per group of at least 100 animals and/or when measured over a period of at least 10 days, which is an amount that is up to or at least 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10%, 15%, 20% or more). The same approach can be taken with other laying animals. Eggs produced by laying hens and other animals are labeled with information indicating the source and/or date of origin.
[127] Also provided by the present invention, in accordance with another embodiment of the first aspect, are one or more eggs, such as an egg carton or egg carton, produced by animals (especially laying hens) that have been treated by a method of according to the first aspect of the present invention. As indicated above, such eggs will normally contain a label indicating their source and/or date of origin. There are also provided downstream products, especially food products, produced from and/or containing eggs or parts thereof produced by animals (especially rearing hens) that have been treated by a method according to the first aspect of the present invention.
[128] An animal feed of a first aspect of the present invention or for use in a first aspect of the present invention may comprise or be supplemented with one or more compounds of the present invention in an amount of 0.001 to 20 g of the one or more compounds per kg of feed, such as 0.002 g/kg to 15 g/kg, or at a level of up to or at least about 0.002 g/kg, 0.005 g/kg, 0.01 g/kg, 0.02 g/kg, 0.03 g/kg, 0.04 g/kg, 0.05 g/kg, 0.1 g/kg, 0.2 g/kg, 0.3 g/kg, 0.4 g/kg, 0 .5 g/kg, 1 g/kg, 2 g/kg, 3 g/kg, 4 g/kg, 5 g/kg, 10 g/kg, 15 g/kg or 20 g/kg. A supply of drinking water for animals or for use in the first aspect of the present invention may comprise or be supplemented with one or more compounds of the present invention in an amount of 0.001 to 20 g of the one or more compounds per L of water, such as 0.002 at 15 g/L, or at a level of up to, or at least about 0.002 g/L, 0.005 g/L, 0.01 g/L, 0.02 g/L, 0.03 g/L, 0.04 g/L, 0.05 g/L, 0.1 g/L, 0.2 g/L, 0.3 g/L, 0.4 g/L, 0.5 g/L, 1 g/L, 2 g/L, 3 g/L, 4 g/L, 5 g/L, 10 g/L, 15 g/L or 20 g/L. The same concentrations may apply to water in which aquatic animals or other animals live.
[129] Optionally, the methods and uses of the present invention are conducted such that, during the course of treatment, the animal ingests and/or absorbs an average daily mean total of FeQ (or an equivalent number of moles of any other substance). or more compounds according to Formula A or B, or other compounds of the invention, as described further in section III.A of this application below) of up to, or at least about 100 μg, 500 μg, 1mg, 10mg, 100mg, 1g, 2g, 3g, 4g, or 5g.
[130] In an additional or alternative option, the methods and uses of the present invention are conducted in such a way that, during the course of treatment, the animal ingests and/or absorbs a total of FeQ (or an equivalent number of moles of any other or more compounds according to Formula A or B, or other compounds of the invention, as described further in section III.A of this application below) of up to, or at least about (a) 5 mg, 10mg, 50mg, 100mg, 500mg, 1g, 5g, 10g, 50g or 100g per individual animal and/or (b) 1mg, 2mg, 3mg, 4mg, 5mg, 10mg 50mg, 100mg, 200mg, 300mg, 400mg, 500mg, 600mg, 700mg, 800mg, 900mg, 1g, 1, 1g, 1, 2g, 13g, 1, 4g, 1.5g, 1.6g, 1.7g, 1.8g, 1.9g, 2g, 2.1g, 2.2g, 2.3g, 2.4g , 2.5g, 2.6g, 2.7g, 2.8g, 2.9g, 3g, 3.5g, 4g, 4.5g, 5g, 6g, 7g , 8g, 9g, 10g, 20g, 30g, 40g, 50g, 60g, 70g, 80g, 90g or 100g per kg of final average body weight as determined on the day of final administration of one or more s compounds.
[131] Accordingly, the present invention also provides animal feed, animal feed supplements, drinking water supply, and ponds (or other contained water-based growing areas) for use in accordance with the present invention and comprising a or more compounds according to Formula A or B, or other compounds of the invention, as described further in section III.A of this application below, at one of the concentrations indicated above. Exemplary animal feeds according to the present invention include feeds for chickens, including (i) starter diets, growth diets and/or finisher diets, in particular for broiler chickens, such as broilers, or (ii) for laying hens, such as pullets or hens, or (iii) for broilers. Also included is food for other poultry, such as a turkey, geese, quail or ducks, or livestock, such as cattle, sheep, goats or swine, alpaca, banteng, bison, camel, cat, deer, dog, donkey, gayal, e.g. zoo animals, game animals, game animals fish (including freshwater and saltwater fish, farmed fish and ornamental fish), other marine and aquatic animals including crustaceans such as, for example, oysters, mussels, clams, shrimp, lobsters, crayfish, crabs, cuttlefish, octopus and squid, domestic animals such as cats and dogs, rodents (such as mice, rats, guinea pigs, hamsters) and horses, as well as any other domestic, wild and farmed animals, including mammals, marine animals, amphibians, birds, reptiles, insects and other invertebrates.
[132] In one embodiment, in the context of the first aspect of the present invention, the animal may be selected from the group consisting of poultry, such as a turkey, geese, quail or ducks, or livestock, such as cattle, sheep, goats or swine, alpaca, banteng, bison, camel, cat, deer, dog, donkey, gayal, e.g. zoo animals, game animals, game animals fish (including freshwater and saltwater fish, ponds and ornamental fish), other marine and aquatic animals, including crustaceans such as oysters, mussels, shellfish, shrimp, lobster, crayfish, crabs, cuttlefish, octopus and squid, domestic animals such as cats and dogs, rodents, ( such as mice, rats, guinea pigs, hamsters) and horses, as well as any other domestic, wild and farmed animals, including mammals, marine animals, amphibians, birds, reptiles, insects and other invertebrates. In a particularly preferred embodiment, the animal is a chicken, for example a meat-like chicken, such as a broiler, or a laying hen, such as a pullet or a broiler or a broiler.
[133] The method of increasing the growth of an animal according to the first aspect of the present invention may be practiced on multiple animals, which may optionally be reared together, and additionally, optionally wherein all animals reared together may be be aged corresponding to within a month, a week or less, such as within 6, 5, 4, 3, 2 or 1 day of each other.
[134] For example, the method can be practiced in a group up to about, or at least 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25, 30, 35, 40, 45, 50, 55, 60, 70, 80, 90, 100, 150, 200, 250, 300, 400, 500, 600, 700, 800, 900, 1x103, 2x103, 3x103, 4x103, 5x103, 6x103 7x103, 8x103, 1x104, 2x104, 3x104, 4x104, 5x104, 6x104, 7x104, 8x104, 9x104, 1x105, 2x105, 3x105, 4x105, 5x105, 6x105, 7x105, 8x105, 9x105, 8x105, 9x105, 1x106 or more, and all animals in the group may optionally have the corresponding age as indicated above. The term "about" in this context can mean within ±50%, ±40%, ±30%, ±20%, ±10%, ±5%, ±4%, ±3%, ±2%, ±1% or less of the indicated value.
[135] Animals treated in accordance with the present invention may be healthy animals, for example animals that are not infected with or disadvantageously colonized by bacteria or other microorganisms. In another embodiment, the animals treated in accordance with the present invention may be unhealthy animals, for example animals that are infected with and/or disadvantageously colonized by bacteria or other microorganisms. An example of disadvantageous bacterial colonization is Campylobacter colonization in the GI tract of chickens; Campylobacter is non-pathogenic and does not cause disease in the chicken itself (although naturally it can lead to food poisoning if present in a downstream meat product produced from the chicken) - however colonization by Campylobacter can be considered disadvantageous for the chicken, as it reduces its ability to grow or use feed effectively. As such, in one embodiment, an animal that is disadvantageously colonized by bacteria or other microorganisms is an animal that exhibits a reduced rate of growth, reduced body weight, reduced weight gain, or a less efficient feed conversion rate due to colonization, compared to a control animal that differs only in that it lacks colonization.
[136] In some embodiments, animals treated in accordance with the present invention may be animals that have been exposed to the litter (including knife matter) of one or more animals of the same or different species. Optionally, the sand may be from unhealthy animals which, for example, animals that are infected and/or colonized disadvantageously by bacteria or other microorganisms. In an embodiment of interest to the present invention, the treated animals may be chickens, such as broilers, and may have been exposed to sand from other chickens, such as dirty sand, as described in the present examples and/or carrying a or more pathogen such as Actinobacillus, Bordetalla, Campylobacter, Clostridium, Corynebacterium, Escherichia coli, Globicatella, Listeria, Mycobacterium, Salmonella, Staphylococcus and Streptococcus. As such, the animals to be treated in accordance with the present invention may be chickens (or other animals) that are infected and/or colonized by one or more of the foregoing pathogens.
[137] Accordingly, in some embodiments, the methods and uses of the present invention may be non-therapeutic in the sense that the animal being treated is healthy and/or the method and use comprise the eventual slaughter of the animal. In other embodiments, the methods and uses of the present invention may include therapeutic benefits to the animals to be treated.
[138] In one embodiment, the methods and uses for increasing the growth of an animal in accordance with the first aspect of the present invention may include increasing one or more characteristics selected from the group consisting of increasing body weight or (in the case of a group of animals), mean body weight (ABW), food intake or (in the case of a group of animals), the mean food intake (AFD), weight gain or (in the case of a group of animals) of animals) the mean weight gain (AWG), the feed conversion index (FCR) and/or the mortality adjusted feed conversion index (MFCR).
[139] In one embodiment (e.g. in the context of a group of chickens grown in a coop) the MFCR over a given period can be calculated as follows: MFCR = period total feed consumption per hen house / ((live weight total chicken house + total weight of dead birds in the chicken house) - total live weight of the chicken house in the previous period)
[140] For example, for a period of 0 to 20 days, the MFCR can be calculated as: MFCR 0 to 20 days = total feed intake 0-20 days/((total body weight on day 20 + mortality weight 0- 20 days)- total body weight (day 0).
[141] The increase in animal growth can be assessed during any convenient period during the animal's growth. It can, for example, be rated from birth to a predetermined time point, such as up to about 10, 20, 30, 40, 50, 60, 70, 80, 90, 100, 110, 120, 130, 140, 150, 160, 170, 180 or more days. The term "about" in this context can mean ±5, ±4, ±3, ±2 or ±1 day. It can be, for example, evaluated from birth to a predetermined time point, such as up to about 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 95% , 96%, 97%, 98%, 99% or 100% of the animal's lifespan. It may alternatively not be measured from birth, but be measured over a period of the animal's life lasting up to about 10, 20, 30, 40, 50, 60, 70, 80, 90, 100, 110, 120, 130, 140, 150, 160, 170, 180 or more days. Again, the term "about" in this context can mean ±5, ±4, ±3, ±2, or ±1 day. It may alternatively not be measured from birth, but be measured over a representative animal life span of about 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90% 95%, 96%, 97%, 98% or 99% of the animal's lifespan.
[142]In the context of using the first aspect of the present invention to increase the growth of broilers, which are typically slaughtered at an average age of 35 days (in the EU) and 47 days (in the US), the increased growth from birth to slaughter age, or can be measured to an earlier age, such as up to 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23 , 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 36, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46 or 47 days. Alternatively, increased growth in broilers cannot be measured from birth, but may be during another period of the broiler's life span that lasts, for example, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 36, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46 or 47 days.
[143] Increased growth may, in some embodiments, refer to an increase in growth in a subject animal compared to a control that is of the same breed of animal as the subject, or an increase in a group of animals in question compared to a control group of an equivalent number of animals of the same breed as the subject group, where the subject and control are of the same age or mean age (ideally within a margin of error of less than one day), in which growth is measured over the same period (ideally within a margin of error of less than one day), and in which the subject and control are created under the same conditions, differing only in that the subject receives one or more compounds from the present invention, in particular one or more compounds according to Formula A or B, or other compounds of the invention as described further in section III.A of this application below, whereas the control does not.
[144] In the context of using the present invention to increase the growth of animals, and in particular poultry, such as broilers and more preferably broilers, an increase in growth rate may constitute a reduction in the subject's MFCR by , up to or at least about 0.01, 0.02, 0.03, 0.04, 0.05, 0.06, 0.07, 0.08, 0.09, 0.10, 0 .11, 0.12, 0.13, 0.14, 0.15, 0.16, 0.17, 0.18, 0.19 or 0.20. The term "about" in this context can include the meaning of ±5x10-3. The reduction in MFCR can, for example, be measured between days 0 to 20 or days 20 to 42 of the life of the animal(s). Under current economic conditions, it can be calculated that a reduction in MFCR of 0.1 will lead to an approximate feed cost savings of about 4 cents per bird over a 42-day growth period and/or about £10 GBP per ton of used pet food. It will be appreciated that these are substantial savings in an industry in which costs are typically controlled to a level of about 0.01 cent per bird.
[145]Furthermore, in the context of using the first aspect of the present invention to increase the growth of animals, and in particular poultry, such as chickens and more preferably broilers, an increase in growth rate may constitute a increase in subject's ABW by up to, or at least about 10 g, 20 g, 30 g, 40 g, 50 g, 60 g, 70 g, 80 g, 90 g, 100 g, 110 g, 120 g, 130g, 140g, 150g, 160g, 170g, 180g, 190g, 200g, 210g, 220g, 230g, 240g, 250g or more. The term "about" in this context can include the meaning of ± 5 g, 4 g, 3 g, 2 g or 1 g. The increase in ABW can, for example, be measured between days 0 to 20 or days 20 to 42 of the life of the animal(s). In the context of animals that normally (i.e. when not treated in accordance with the present invention) have a higher ABW than the normal ABW of broilers (i.e. when not treated in accordance with the present invention), then previous values can be increased proportionally. That is, for example, in the case of an animal that has a normal ABW 10 times greater than the normal ABW of a broiler, then the increase in growth rate provided by the present invention may constitute an increase in the subject's ABW up to , or at least about 100g, 200g, 300g, 400g, 500g, 600g, 700g, 800g, 900g, 1000g, 1100g, 1200g, 1300g, 1400g, 1500g, 1600g, 1700g, 1800g, 1900g, 2000g, 2100g, 2200g, 2300g, 2400g, 2500g or more, wherein the term "about" in this context may include the meaning of ± 50 g, 40 g, 30 g, 20 g or 10 g.
[146]Furthermore, in the context of using the first aspect of the present invention to increase the growth of animals, and in particular poultry, such as chickens and more preferably broilers, an increase in growth rate may constitute a increase in subject's mean weight gain (AWG) up to or at least about 10 g, 20 g, 30 g, 40 g, 50 g, 60 g, 70 g, 80 g, 90 g, 100 g, 110 g, 120g, 130g, 140g, 150g, 160g, 170g, 180g, 190g, 200g, 210g, 220g, 230g, 240g, 250g, 260g, 270g, 280g, 290 g, 300 g or more during a period of growth, compared to a control animal or group of animals. The term "about" in this context can include the meaning of ± 5 g, 4 g, 3 g, 2 g or 1 g. The increase in AWG can, for example, be measured between days 0 to 20 or days 20 to 42 of the life of the animal(s), or over a period of time selected from 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 36, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46 or 47 days. In the context of animals that normally (i.e. when not treated in accordance with the present invention) have a higher ABW than the normal ABW of broilers (i.e. when not treated in accordance with the present invention), then previous values can be increased proportionally. That is, for example, in the case of an animal that has a normal ABW 10 times greater than the normal ABW of a broiler, for an equivalent period, then the increase in growth rate provided by the present invention may constitute an increase in the subject's ABW up to or at least about 100g, 200g, 300g, 400g, 500g, 600g, 700g, 800g, 900g, 1000g, 1100g, 1200g, 1300g , 1400g, 1500g, 1600g, 1700g, 1800g, 1900g, 2000g, 2100g, 2200g, 2300g, 2400g, 2500g, 2600g, 2700g, 2800g, 2900g, 3000 or more, wherein the term "fence" in this context may include the meaning of ±50g, 40g, 30g, 20g or 10g.
[147] Prior to the present invention, in the US, the average age at slaughter of a broiler is 47 days with an average weight of 2.6 kg; at the age of 42 days the average weight can be around 2.5 kg, and in the EU the average slaughter age of a broiler is 35 days with an average weight of 2.1-2.2 kg. It will be appreciated that, as a result of the increased growth provided by the methods and uses of the present invention, it will be possible to reach the target weight and harvest the animal or animal products at an earlier stage in the animal's life than would be possible with a control. For example, in the context of a broiler, it may be possible to slaughter the animal after reaching a target body weight of 1, 2, 3, 4, 5, 6, 7, 8, 9, 10 or more days earlier than a control. In this context, the target body weight of a broiler can be, can be up to, or can be at least about 1000g, 1100g, 1200g, 1300g, 1400g, 1500g, 1600g, 1700g , 1800g, 1900g, 2000g, 2100g, 2200g, 2300g, 2400g, 2500g, 2600g, 2700g, 2800g, 2900g, 3000g, 3100g, 3200g, 3300g, 3400 g, 3500 g or more. The term "about" in this context may include ±50g, ±40g, ±30g, 20g, or ±±10g of the indicated value. In other words, broilers can be slaughtered at 47, 46, 45, 44, 43, 42, 41, 40, 39, 38, 37, 36, 35, 34, 33, 32, 31, 30, 29, 28, 27, 26 or 25 days, ideally when it has reached a target body weight at the time of slaughter. Thus, for example, in one embodiment of the present invention, the broiler is raised to a target weight of about 2.6 kg and the method or use includes the step of slaughtering the animal after it has reached a target body weight 1, 2, 3, 4, 5, 6, 7, 8, 9, 10 or more days before the age of 47 days. In an exemplary embodiment, the broiler is raised to a target weight of about 2.5 kg and the method or use includes the step of slaughtering the animal after it has reached a target body weight of 1, 2, 3, 4, 5 , 6, 7, 8, 9, 10 or more days before the age of 42 days. In another exemplary embodiment, the broiler is raised to a target weight of about 2.2 kg and the method or use includes the step of slaughtering the animal after it has reached a target body weight of 1, 2, 3, 4, 5, 6, 7, 8, 9, 10 or more days before the age of 35 days.
[148] In another embodiment, the animal is reared for the same amount of time as the industry standard, but has a higher body weight (such as about, at least, or up to 0.1%, 0.5% , 1%, 2%, 3%, 4%, 5%, 10%, 15%, 20%, 25% or more) than the industry standard at the end of the build process. Thus, in the context of broilers, the animal can be slaughtered at a weight of about 1000 g, 1100 g, 1200 g, 1300 g, 1400 g, 1500 g, 1600 g, 1700 g, 1800 g, 1900 g, 2000g, 2100g, 2200g, 2300g, 2400g, 2500g, 2600g, 2700g, 2800g, 2900g, 3000g, 3100g, 3200g, 3300g, 3400g, 3500g or more, where at the time of slaughter, the body weight is at least or up to 0.1%, 0.5%, 1%, 2%, 3%, 4%, 5%, 10%, 15%, 20%, 25% or more than the control. The term "about" as applied to weight in that context may include ±50g, ±40g, ±30g, ±20g or ±10g of the indicated value.
[149] In yet another embodiment, as a result of the increased growth effect provided by the methods and uses of the first aspect of the present invention, the animal is able to utilize animal feed more efficiently than a control. Accordingly, in another embodiment, the methods and uses of the present invention include the option of raising an animal to reach a target body weight using less animal feed than is necessary for a control to reach the target weight. For example, it may be possible to use the present invention to raise an animal to reach the target weight using an amount of animal feed that is reduced in weight by 0.01%, 0.05%, 0.1%, 0.2 %, 0.3%, 0.4%, 0.5%, 1%, 2%, 3%, 4%, 5%, 10%, 15%, 20%, 25% or more compared to the amount of the same animal feed needed by a control to achieve the same target weight. In this context, the target body weight of a broiler can be, can be up to, or can be at least about 1000g, 1100g, 1200g, 1300g, 1400g, 1500g, 1600g, 1700g , 1800g, 1900g, 2000g, 2100g, 2200g, 2300g, 2400g, 2500g, 2600g, 2700g, 2800g, 2900g, 3000g, 3100g, 3200g, 3300g, 3400 g, 3500 g or more. The term "about" in this context may include ±50g, ±40g, ±30g, ±20g or ±10g of the indicated value.
[150] For example, in the context of the industry standard for raising broilers for 42 days, it is typical to provide each chicken with a total of 5.2 kg of feed over its lifetime (an average of 123.8 g of food per day of life). In such a situation, in one embodiment, the present invention involves feeding the chicken a total amount of chicken feed that is reduced by 5.2 kg and/or reduced by an average of 123.8 g of feed per day, 0 .01%, 0.05%, 0.1%, 0.2%, 0.3%, 0.4%, 0.5%, 1%, 2%, 3%, 4%, 5%, 10 %, 15%, 20%, 25% or more during its creation.
[151] Accordingly, the methods and uses of the present invention may further comprise the step of rearing the animal to allow for increased growth.
[152] Another embodiment according to the first aspect of the present invention provides a method for preventing or reducing colonization of the gastrointestinal tract of an animal (such as an animal described above) with Campylobacter and/or other bacterial agents or microorganisms by making causing the animal to ingest and/or absorb an effective amount of one or more compounds having the structure of Formula A or B, or other compounds of the invention as described further in section III.A of this application below. In particular, it relates to reducing or preventing colonization of the gastrointestinal tract of poultry (such as poultry types as described above) with Campylobacter. It also relates to uses of one or more compounds having the structure of Formula A or B or other compounds of the invention as described further in section III.A of this application below to prevent bacteria from adhering to the gastrointestinal tract wall of animals and to treat or prevent Campylobacter infection and/or other bacteria or microorganisms in humans and animals.
[153] Accordingly, in another embodiment of the first aspect of the present invention, there is provided a method for disinfecting an animal comprising administering to said animal at least one or more compounds having the structure of Formula A or B or other compounds of the invention as described further in section III.A of this application below, in an amount effective to reduce the number of Campylobacter and/or other microorganisms or bacteria present in the gastrointestinal tract of said animal.
[154] Another embodiment of the first aspect of the present invention also provides a method for disinfecting an animal comprising administering to said animal at least one or more compounds having the structure of Formula A or B, or other compounds of the invention, as described further in Section III.A of this application below, in an amount effective to prevent said Campylobacter and/or other microorganisms or bacteria from forming a biofilm in the gastrointestinal tract of said animal or to reduce the amount of biofilm formed by Campylobacter and/or other bacteria or microorganisms in the intestinal tract of said animal.
[155] Another embodiment of the first aspect of the present invention also provides a method of preventing or reducing the transmission of Campylobacter infection, and/or infection by other bacteria or microorganisms, from one animal to another, for example by preventing or reducing the spread of Campylobacter and/or infection by other bacteria or microorganisms, within a herd or herd of animals, for example by preventing the spread of Campylobacter infection and/or infection by other bacteria or microorganisms, within a herd of animals chickens, including broilers; said method comprising administering to said animals, for example said flock or flock of animals, for example said flock of chickens, one or more compounds having the structure of Formula A or B, or other compounds of the invention as described further further in section III. The one of this application below in an amount effective to prevent Campylobacter and/or other bacteria or microorganisms from forming a biofilm in the gastrointestinal tract of said animal or to reduce the amount of biofilm formed by Campylobacter and/or other bacteria or microorganisms, in the intestinal tract of said animal.
[156] These methods can allow disinfection, prevention of biofilm formation and reduction of Campylobacter and/or other bacteria or microorganisms among animals, preventing or reducing Campylobacter and/or other bacteria or microorganisms from the intestinal tract. gastrointestinal tract of said animals. This is advantageous because the less Campylobacter and/or other bacteria or microorganisms that are in an animal's gastrointestinal tract at the time of slaughter, the lower the risk of contamination of the animal's meat with Campylobacter and/or other bacteria or microorganisms. . The less Campylobacter and/or other bacteria or microorganisms that are in an animal's gastrointestinal tract, the less chance of Campylobacter and/or other bacteria or microorganisms forming a biofilm in the animal's gastrointestinal tract. The less Campylobacter and/or other bacteria or microorganisms that are in an animal's gastrointestinal tract, the less chance that Campylobacter and/or other bacteria or microorganisms will disperse from one animal to another, for example, within a herd or herd of animals.
[157] These methods can also be used to reduce the amount of colonization of the gastrointestinal tract of any animal with Campylobacter and/or other bacteria or microorganisms. It may be particularly advantageous to provide one or more compounds having the structure of Formula A or B, or other compounds of the invention as described further in section III.A of this application below for animals that will be slaughtered for human consumption. Poultry includes birds that are used for human consumption, such as chickens, geese, turkeys and ducks. It is particularly advantageous to use the compounds of the present invention to reduce or prevent colonization of the gastrointestinal tract of poultry, in particular chickens, and more particularly broilers, laying hens and/or broilers, with Campylobacter and/or other bacteria or microorganisms because chickens are the main source of human infection with Campylobacter.
[158] The number of Campylobacter and/or other bacteria or microorganisms in the gastrointestinal tracts of animals can be reduced by the methods of the present invention. In one embodiment, the number of colony forming units (cfu) of Campylobacter and/or other bacteria or microorganisms in the gastrointestinal tract of an animal treated with the compounds of the present invention can be reduced by 50%, by 60%, by 70%, 80%, 90% or 100%. In one embodiment, Campylobacter and/or other bacteria or microorganisms can be substantially eradicated from the gastrointestinal tract of animals treated by the method of the present invention.
[159] 10,000 cfu of Campylobacter is sufficient for successful colonization of the chicken. 1,000 cfu of Campylobacter is enough to infect a human and cause disease in a human. Therefore, an effective amount of a compound of the present invention is sufficient of the compound to reduce the number of Campylobacter and/or other bacteria or microorganisms in the gastrointestinal tract of an animal to a number that is unlikely to cause infection in humans, such as less than 10,000 cfu, 5,000 cfu, 1,000 cfu, 500 cfu, 400 cfu, 300 cfu, 200 cfu, 100 cfu, 90 cfu, 80 cfu, 70 cfu, 60 cfu, 50 cfu or less. The number of cfu of Campylobacter/or other bacteria or microorganisms that would be ingested by a human if they ate meat from an infected animal may be related to the number of Campylobacter and/or other bacteria or microorganisms in the animal's gastrointestinal tract in the time of slaughter, but also depends on other factors, such as the amount of contamination of the meat with the contents of the animal's gastrointestinal tract at the time of slaughter.
[160] An effective amount of one or more compounds having the structure of Formula A or B, or other compounds of the invention, as described further in Section III.A of this application below, in this context, may be an amount which is sufficient of one or more compounds to prevent colonization of the animal's gastrointestinal tract with Campylobacter and/or other bacteria or microorganisms.
[161] In one embodiment, one or more compounds having the structure of Formula A or B, or other compounds of the invention as described further in Section III.A of this application below can cause Campylobacter and/or other bacteria or microorganisms less virulent and less capable of infecting humans, even if the total number of Campylobacter and/or other bacteria or microorganisms in the gastrointestinal tract does not decrease. In this embodiment, administering a compound of the present invention to an animal may affect the metabolism of Campylobacter and/or other bacteria or microorganisms and make them less adaptable to the environment (e.g., less mobile) so that they cannot colonize. the gastrointestinal tract and are less likely to be transmitted to other animals or to humans.
[162] An effective amount of one or more compounds given to an animal should be sufficient to provide the required degree of reduction of Campylobacter and/or other bacteria or microbial colonization. This may depend on the type of compost and/or the size of the animal.
[163] In one embodiment, the one or more compounds may be provided in a pet food, pet drink, or other compositions of concentration within the range of about 1μM to about 1M, preferably greater than 10 μM, 20μM, 30μm, 40 μm, 50 μm, 70 μm, 80 μm, 90 μm, 100 μm, 110 μm, 120μm, 130μm, 140 μm, 150 μm, 160 μm, 170 μm, 180 μm, 190 μm, 200 μm , 250 μM, 300 μM, 350 μM, 500 μM, 1 mM or more.
[164] For example, the concentration of one or more compounds may be: (a) up to 1 μM, 2μM, 3μM, 4μM, 5μM, 10μM, 15 μM, 20 μM, 25 μM, 30 μM; (b) within a selected group range consisting of 35 to 335 μM, 40 to 300 μM, 50 to 300 μM, 50 to 250 μM, 50 to 200 μM, 60 to 300 μM, 60 to 250 μM, 60 to 200 µM, 80 to 300 µM, 80 to 250 µM, 80 to 200 µM, 100 to 300 µM, 100 to 250 µM, or 100 to 200 µM; or (c) at least, or about, 345 µM, 350 µM, 360 µM, 370 µM, 380 µM, 390 µM, 400 µM, 450 µM, 0.5 mM, 1 mM, 2 mM or more.
[165] In another embodiment, the concentration may be within a range selected from the group consisting of about 1μM to about 1mM, or about 30μM to about 0.5mM, or about 60μM to about 0.3 mM.
[166]In the case of animal beverage (such as drinking water) or other types of composition, optionally, the concentration of one or more compounds in the composition may be within the range of 0.002 to 15 g/L, or at a level of, up to or at least about 0.002 g/L, 0.005 g/L, 0.01 g/L, 0.02 g/L, 0.03 g/L, 0.04 g/L, 0.05 g /L, 0.1 g/L, 0.2 g/L, 0.3 g/L, 0.4 g/L, 0.5 g/L, 1 g/L, 2 g/L, 3 g /L, 4 g/L, 5 g/L, 10 g/L, 15 g/L or 20 g/L.
[167] In another embodiment, one of the further compounds may be provided in a pet food, pet drink or other composition in a unit dosage formulation, and/or in a concentration to deliver up to, or at least about, about 1ng, 10ng, 50ng, 100ng, 500ng, 1µg, 10µg, 50µg, 100µg, 500µg, 1mg, 10mg, 100mg, 500mg, 1g, 2g, 3g, 4 g, or 5 g of one or more compounds.
[168] The methods and uses of the present invention may further comprise the step of harvesting a product from the animal reared with increased growth.
[169] The harvested product may be the body or body part of the animal. In that case, the harvesting process includes the step of slaughtering the animal and, optionally, preparing an animal carcass or part thereof as a product, such as a meat product. Accordingly, the harvested body or body part of the animal thereafter may be a non-food product, a food product or a precursor of a food product. Carcasses and carcass parts can undergo a process known as rendering to be transformed into food, fats and other human and non-human materials that can be sold to produce commercial products such as cosmetics, paints, cleaning products, polishes, glues, soaps and paints. In addition, such products that may be foodstuffs include, but are not limited to, blood, bone, including bone, bone meal, etc., broths and solutions created with animal fat, bone and/or connective tissue, carmine also known as cochineal (food color), casein (found in milk and cheese), civet oil (food flavoring additive), gelatin, ichthyocola (which can, for example, be used in clarifying beer and wine), L-cysteine (which it can for example be used in the production of biscuits and bread), lard, meat (including fish, poultry and game) and rennet (commonly used in the production of cheese). Meat and products may be of particular interest.
[170] In an embodiment of particular interest in the context of the present invention, the animal is a chicken, for example, a meat-type chicken, such as a broiler, or a laying hen, such as a pullet or a hen, and the product is harvested from the bred animal. More preferably, the animal is a broiler chicken, such as broiler chicken, and the harvested product is a carcass or part of the carcass of the chicken. After slaughtering to produce the carcass, it may or may not be further processed so as to remove one or more selected items from the group consisting of feathers, offal, skin from the neck, head, legs and other items and may produce a set of carcass ready for sale as a meat product, or ready to ship for further processing. In one embodiment, the processed carcass may retain the neck or neck skin, or at least 50%, 60%, 70%, 80%, 90% or more thereof, as determined by length or weight. The average neck or neck skin weight may be in the range of 15-25g further processing may include performing a shearing operation where the carcass is cut into individual pieces and may involve deboning (i.e. where the bones are removed of specific parts) to produce items such as breast fillets or other boneless products.
[171] In an exemplary embodiment, a process for slaughtering and/or processing a chicken may include any one or more of the following methodological steps: i) the birds arrive at the processing facility, typically in plastic crates; (ii) blue light is used to calm birds; (iii) the birds are hung; (iv) birds enter a stunning tank; (v) the birds are slaughtered using a neck bleed, optionally with a delay support for the bleed of the birds; (vi) the skin and/or feathers of birds are heated, for example with water, to loosen the pores holding the feathers; (vii) feathers are removed, for example, using rubber fingers; (viii) an inspection is conducted to remove any birds that fail a quality control assessment; (ix) a drill or other implement is used to create a hole in the housing and remove the anus; (x) removing the intestines and other internal organs, typically through the previously created hole; (xi) optionally, the production line is divided for the production of whole chickens and chicken parts; (xii) chicken pieces can be cut using an automated process and through manual labor (cutting workers); optionally including liver, kidney and/or separate hearts; (xiii) the entire chicken carcass and/or chicken parts can be labeled directly on the processing plant floor, ready for the grocery store (optionally including additional pricing) so the product can go directly to the store shelf.
[172] It will be appreciated that alternative methods of stunning birds are available and may be used in place of the method indicated in the preceding method and/or used more generally in accordance with the first aspect of the present invention. Examples of alternative methods of stunning birds include, for example, controlled atmosphere stunning, controlled atmosphere killing, biphasic CO2 and slow controlled decompression.
[173] Controlled atmosphere stunning (also known as gas stunning) can be applied to birds in transport crates, which can be transported through a tunnel or other chamber filled with increasing concentrations of carbon dioxide, inert gases (argon or nitrogen) or a mixture of these gases. The gas or gases induce unconsciousness before slaughter. For example, at this point, birds are hung up in shackles, while unconscious, and transported to the killing machine for slaughter.
[174]Controlled atmosphere killing (CAK) can be operated by exposing birds to lethal concentrations of gases long enough that they are actually killed rather than stunned (to avoid the risk of birds regaining consciousness after exit the gaseous atmosphere). For example, carbon dioxide directly depresses the central nervous system and produces rapid unconsciousness. However, carbon dioxide is aversive to chickens (generally if levels are above 20%). Inhalation of inert gases (eg, argon and nitrogen) can also be used, when inhaled in high concentrations, to cause oxygen deprivation in the body, leading to death.
[175] Two-phase CO2 is a newer method of gas stunning that uses carbon dioxide in two stages to kill poultry. The first stage containing up to 40% carbon dioxide (only moderately aversive to chickens), renders the birds unconscious, the second stage follows with lethal carbon dioxide levels.
[176]Slow controlled decompression may include the use of a Low Atmospheric Pressure System (LAPS). Death by LASP mimics the physiological effects of ascending to high altitudes using slow controlled decompression, which allows the bird's body to adjust to changes in pressure and thus lose consciousness (due to lack of oxygen) with minimal discomfort.
[177]Alternatively, the bird cannot be stunned prior to slaughter, for example in the case of production of a meat product in accordance with religious laws such as the Halal, Qurrbani/Udhia and/or Shechita slaughter laws.
[178] Carcass processing can be conducted at sufficiently low refrigeration temperatures, such as about 1, 2, 3, 4 or 5 °C.
[179] Consequently, after processing the animal carcass and/or producing parts thereof, the carcass or part thereof may be further processed to produce a value-added product and this may include one or more steps necessary to prepare a ready-to-eat product, which may include the addition of one or more seasoning, breading, sauces and marinades, as well as special packaging to meet market demands for convenient products.
[180]Additionally, or alternatively, the harvested product may, for example, be a by-product of the animal, such as milk, eggs, wool, hair, feathers or litter or other faecal matter and may be collected from the animal without the need for slaughter. the animal. Such harvested products can then be further processed and converted into other products. For example, in the context of milk, more dairy products can be produced (such as butter, cheese, curds, yogurt, whey, powdered milk, cream, spreads and other cultured dairy foods, frozen desserts such as ice cream, other frozen desserts made with dairy ingredients). In the context of eggs, other products (in particular food products) may be produced which contain or are produced with all or part of the eggs collected. In the case of wool, hair or feathers, it may, for example, be possible to produce fibers or fabrics, products containing wool, hairs or feathers (such as upholstery products) or the products may be chemical or enzymatic processes of the wool, hair or feathers. . For example, amino acids can be produced as a breakdown product of wool, fur or feathers. Chicken litter can include a mixture of faeces, food waste, bedding materials and feathers that can be recycled or composted and then spread over arable land as a low-cost organic fertilizer.
[181] Any and all stages of the entire animal husbandry process, animal harvesting, animal slaughter, carcass processes, animal product production, food production, packaging, labelling, shipping, storage and sale in accordance with the first aspect of the present invention may benefit from applying a surface or coating disinfection in accordance with the third aspect of the present invention, as discussed further below. For example, animal husbandry areas according to the first aspect of the present invention may contain one or more disinfected surfaces obtained using the methods, uses and compositions of the third aspect of the present invention. Containers for transporting animals in accordance with the practice of the first aspect of the present invention may contain one or more disinfected surfaces obtained using the methods, uses and compositions of the third aspect of the present invention. The apparatus used in the slaughter of animals in accordance with the practice of the first aspect of the present invention may contain one or more disinfected surfaces obtained using the methods, uses and compositions of the third aspect of the present invention. Apparatus used in the processing and/or labeling of an animal carcass or a part thereof, in accordance with the practice of the first aspect of the present invention, may have one or more disinfected surfaces obtained using the methods, uses and compositions of the third party. aspect of the present invention. The animal product, including a carcass, a meat product or any other animal product, as produced in accordance with the first aspect of the present invention, can be disinfected using the methods, uses and compositions of the third aspect of the present invention. The package, containers and/or package for containing an animal product, including a carcass, a meat product or any other animal product as produced in accordance with the first aspect of the present invention, can be disinfected using the methods, uses and compositions of the third aspect of the present invention. These combinations of the approaches presented by the first and third aspects of the present invention all form optional embodiments of the first aspect of the present invention.
[182] The present invention also provides products produced by, and/or harvested from, animals treated in accordance with the first aspect of the present invention, including any and all of the products discussed above, and downstream products including or produced therefrom.
[183] For example, the present application provides a meat or meat product produced in accordance with the present invention. For example, it may provide a carcass or part thereof that is of a greater weight than a standard carcass or part thereof, or is from an animal that is younger than a control. Additionally, or alternatively, the carcass or part thereof, or any other product obtained from the animal, may have a reduced level of microbial (such as bacterial, including Campylobacter) infection or colonization and/or a reduced incidence of biofilms thereon, compared to a control.
[184] It will be appreciated that the above methods and uses for increasing the growth of an animal may also be applied to humans, for example, to increase the growth of humans (such as an aid to body mass development) and/or improve the efficiency or FCR with which humans digest food. This could, for example, have applications for military personnel in helping to reduce the load of food transport and/or assisting in cases of food shortages, increasing the dietary benefit of available food. B. Enhancing the effect of antibiotics and other antimicrobial agents and addressing antibiotic resistance
[185] Compounds having the structure of Formula A or B, or other compounds of the present invention as described further in section III.A of this application, have been found to be particularly useful in treating or preventing infection by resistant microorganisms. to antibiotics. The compounds can be administered in order to cause the microorganisms to lose their antibiotic resistance.
[186] In Example 9, it was shown that a kanamycin resistant strain of E. coli did not grow when treated with Fe-QA and kanamycin. However, administration of Fe-QA alone had no impact on strain growth. In Example 10, it was shown that the growth of another kanamycin resistant bacterial strain, Campylobacter, was retarded when it was treated with Fe-QA and kanamycin. In Example 14, it was shown that the growth of an antibiotic resistant clinical isolate of Pseudomonas was also retarded when treated with Fe-QA (also known as FeQ) and kanamycin. The effect, therefore, is not just limited to the bacterium, E. coli. In Example 12, a gentamicin-resistant Enteropathogenic E. coli (EPEC) wild-type strain was also shown to lose its resistance when treated with a combination of Fe-QA and gentamicin. The example demonstrates that the effect is not limited to kanamycin, but is seen with other antibiotics. Furthermore, the effect is not limited to the compound, Fe-QA, but is also seen with the other compounds. In Example 11, a gentamicin-resistant Enteropathogenic E. coli (EPEC) wild-type strain was also shown to lose its resistance when treated with a combination of Fe-Tyr and gentamicin. Thus, the compounds are capable of causing antibiotic-resistant bacteria to lose their resistance, and therefore, the administration of the compounds and antibiotics can be used to treat antibiotic-resistant microorganisms (or prevent infection by these microorganisms).
[187] Therefore, a second aspect of the present invention is based on the surprising finding that compounds having the structure of Formula A or B, or other compounds of the present invention, described in section III.A of this application, can be used for increase the microorganism's sensitivity to antimicrobial agents, potentiate the effect of antibiotics and other antimicrobial agents, and address antimicrobial and antibiotic resistance.
[188] In another preferred option of the second aspect of the present invention, one or more compounds are selected from the group consisting of a complex of an amino acid with Fe III, and a complex of an ahydroxy acid with Fe III, or salts and/or hydrates from them. In particularly preferred options of the second aspect of the present invention, one or more compounds may or may not be selected from any one or more of the group consisting of a complex of quinic acid with Fe III (such as a complex with the structure of Formula IX) , a complex of L-tyrosine with Fe III (such as a complex having the structure of Formula VIII), a complex of L-DOPA with Fe III (such as a complex having the structure of Formula VII), and a complex of L- phenylalanine with Fe III. Accordingly, in an embodiment of the second aspect of the invention, a complex of L-tyrosine with Fe III (such as a complex with the structure of Formula VIII) is particularly preferred. Optionally, one or more compounds is not a complex of quinic acid with Fe III (such as a complex having the structure of Formula IX).
[189] In a particularly preferred embodiment, compounds having the structure of Formula A or B, or other compounds of the present invention as described in section III.A of this application can be used in combination with antimicrobial agents to treat or prevent infection. by antibiotic-resistant bacteria including Streptococcus pneumoniae, Campylobacter, Neisseria gonorrhoeae, Salmonella (including non-typhoid drug-resistant Salmonella and drug-resistant Salmonella typhi serotype), Methicillin-resistant Staphylococcus aureus (MRSA), Shigella, Vancomycin-resistant Enterococcus (VRE) , Vancomycin-resistant Staphylococcus aureus (VRSA), Erythromycin-resistant Group A Streptococcus, Clindamycin-resistant Group B Streptococcus, Carbapenem-resistant Enterobacteriaceae (CRE), drug-resistant tuberculosis, extended-spectrum Enterobacteriaceae (ESBL), Acinetobacter-resistant multidrug (including MRAB), Clostridium difficil and, Enteropathogenic E. coli (EPEC), Pseudomonas aeruginosa, and Uropathogenic E. coli (UPEC). In another preferred embodiment. In another embodiment, the compounds can be used in combination with antimicrobial agents to treat or prevent infection by antibiotic-resistant bacteria including S. epidermidis, E. faecalis, E. coli, S. aureus, Enteropathogenic Escherichia coli (EPEC), Uropathogenic Escherichia coli (UPEC), Pseudomonas, Streptococcus anginosus, Salmonella, including Salmonella Enteritidis and Salmonella Typhimurium, Mycoplasma, Eimeria, Enterococcus, Brachyspira and Clostridium perfringen. In a preferred embodiment, the antimicrobial compounds and agents can be administered as a pharmaceutical composition or a food additive.
[190] Antibiotic-resistant microorganisms (and other microorganisms resistant to other forms of antimicrobial agent) can be treated with one or more compounds and one or more antibiotics or other antimicrobial agents separately, sequentially, or simultaneously. Preferably, one or more compounds are administered at the same time as one or more antibiotics or other antimicrobial agents, or preferably such that the compounds and the antibiotic(s) are present at the same time. (The compounds and the antibiotics/antimicrobial agents can therefore also be administered sequentially.) As described above, the compounds can also be formed in vivo. In this case, the precursors can be administered with the antibiotics or other antimicrobial agents. For example, antibiotics or other antimicrobial agents can be administered with ferrous sulfate and tyrosine (which form Fe-Tyr in vivo) or ferrous sulfate and L-DOPA (which form Fe-DOPA in vivo), or ferrous sulfate and L-phenylalanine. (which form Fe-Phe in vivo).
[191]Combinations of the compounds and the antibiotic(s) or other antimicrobial agents may be used to treat any infections, including, but not limited to, the following infections: acute bacterial skin infections, hospital-acquired bacterial pneumonia, ventilator-acquired bacterial pneumonia, urinary tract infections, abdominal infections, kidney infections, gonorrhea, osteomyelitis, lung infections, and respiratory tract infections.
[192] The compounds may also be used in combination with antibiotics or other antimicrobial agents to allow lower doses of antibiotic or other antimicrobial agents to be used to treat not only antibiotic-resistant microorganisms (and/or other antibiotic-resistant microorganisms) other forms of antimicrobial agent), but also for the treatment of microorganisms that are not resistant to antibiotics or other antimicrobial agents. In other words, the compounds can allow lower doses of an antibiotic or other antimicrobial agent to be used to treat or prevent infections and can also be administered to patients and animals prophylactically. For example, the compounds can be administered to poultry prophylactically so that a lower dose of antibiotic and/or other antimicrobial agent is needed to treat the birds if they become infected.
[193] Accordingly, a second aspect of the present invention provides a method for the treatment or prophylaxis of a microbial infection or colonization in a patient or animal, the method comprising administering to the patient or animal a product selected from the group that consists of a pharmaceutical agent or veterinary product, a medical device or a dietary product, wherein the product comprises one or more compounds having the structure of Formula A or B, or other compounds of the invention as described further in section III.A hereof. application below, and preferably wherein the pharmaceutical or veterinary product, medical device or dietary product is administered to the patient or animal separately, simultaneously or sequentially with the administration of one or more antimicrobials and/or antibiotics.
[194] In other words, the second aspect of the present invention provides a pharmaceutical or veterinary product, a medical device or a dietary product, wherein the product comprises one or more compounds having the structure of Formula A or B, or other compounds of the invention as further described in section III.A of this application below, for use in a method of treating or prophylaxis of a microbial infection or colonization in a patient or animal, preferably wherein, during use, the pharmaceutical or veterinary product, the medical device or dietary product is administered to the patient or animal separately, simultaneously or sequentially with the administration of one or more antimicrobials and/or antibiotics.
[195] Likewise, the second aspect of the present invention also provides one or more antimicrobials and/or antibiotics, for use in a method of treating or prophylaxis of a microbial infection or colonization in a patient or animal, preferably wherein, during use, the pharmaceutical or veterinary product, medical device or dietetic product is administered to the patient or animal separately, simultaneously or sequentially with the administration of a pharmaceutical or veterinary product, a medical device or a dietetic product, wherein the The product comprises one or more compounds having the structure of Formula A or B, or other compounds of the invention as described further in section III.A of this application below.
[196]Microbial infection or colonization in a patient or animal can, for example, be pathogenic or non-pathogenic microbes. Non-pathogenic microbes can, for example, cause colonization of a host without causing or producing any disease or disturbance of the host. The microbial infection or colonization directed by the second aspect of the present invention may be prokaryotic. Examples of prokaryotic microbes include bacteria and archaea. The microbial infection or colonization directed by the second aspect of the present invention may be eukaryotic. Examples of eukaryotic microbes include protists (such as algae and slime fungi), fungi, multicellular micro-animals, and plants including green algae.
[197] A class of microbes of particular interest for the application of the second aspect of the present invention are bacteria, including pathogenic and non-pathogenic bacteria. By way of various non-limiting examples, bacteria of particular interest for the application of the second aspect of the present invention include gram positive bacteria, gram negative bacteria, biofilm forming bacteria, extracellular bacteria, intracellular bacteria (including facultative and obligate intracellular bacteria), aerobic bacteria and anaerobic bacteria. Some bacterial genera of interest, without limitation, include Bacillus, Bartonella, Bordetella, Borrelia, Brucella, Campylobacter, Chlamydia and Chlamydophila, Clostridium, Corynebacterium, Enterococcus, Escherichia, Francisella, Haemophilus, Helicobacter, Legionella, Leptospira, Listeria, Mycobacterium, Mycoplasma, Neisseria, Pseudomonas, Rickettsia, Salmonella, Shigella, Staphylococcus, Streptococcus, Treponema, Ureaplasma, Vibrio, and Yersinia. Some bacterial species of interest, without limitation, include Bacillus anthracis, Bacillus cereus, Bartonella henselae, Bartonella quintana, Bordetella pertussis, Borrelia burgdorferi, Borrelia garinii, Borrelia afzelii, Borrelia recurrentis, Brucella abortus, Brucella canis, Brucella melitensis, Brucella suis, Campylobacter jejuni, Chlamydia pneumonia, Chlamydia trachomatis, Chlamydophila psittaci, Clostridium botulinum, Clostridium difficile, Clostridium perfringens, Clostridium tetani, Corynebacterium diphtheria, Enterococcus faecalis, Enterococcus faecium, Escherichia coli, Francisella tularensis, Haemophilus influenza, Helicobacter pylori, Legionella pneumophila, Leptospira interrogans, Leptospira interrogans, Leptospira interrogans, Leptospira interrogans, Leptospira interrogans Leptospira santarosai, Leptospira weilii, Leptospira noguchii, Listeria monocytogenes, Mycobacterium leprae, Mycobacterium tuberculosis, Mycobacterium ulcerans, Mycoplasma pneumonia, Neisseria gonorrhoeae, Neisseria meningitides, Pseudomonas aeruginosa, Rickettsia rickettsia, Salmonella typhi, Salmonella typhimurium, Shigella sonnei, Staphylococcus aureus, Staphylococcus epidermidis, Staphylococcus saprophyticus, Streptococcus agalactiae, Streptococcus pneumonia, Streptococcus pyogenes, Treponema pallidum, Ureaplasma urealyticum, Vibrio cholera, Yersinia pestis, Yersinia enterocolitica, Yersinia pseudotuberculosis.
[198] The treatment or prophylaxis of the second aspect of the present invention may be directed to one or more microorganisms that have increased resistance or tolerance to one or more antimicrobial agents. For example, the one or microorganisms may be, or include, one or more antibiotic-resistant bacteria.
[199] As such, in the embodiment where the second aspect of the present invention is carried out by administering separately, simultaneously or sequentially with the administration of one or more antimicrobials and/or antibiotics, then some or all of the one or more antimicrobials and/or antibiotics can be antimicrobials and/or antibiotics to which the microorganisms to be fought are resistant. In other words, typically the microorganisms to be combated can be those in which, in the absence of the product, they comprise one or more compounds having the structure of Formula A or B, or other compounds of the invention as described further in section III. As of this application below, one or more microorganisms is/are resistant to the one or more antimicrobials and/or antibiotics administered to the patient or animal. Antimicrobial resistance can include the meaning of resistance of a microorganism to an antimicrobial drug that was originally effective for the treatment of infections caused by it. Resistant microorganisms are able to withstand the onslaught of antimicrobial drugs, such as antibacterial (e.g. antibiotics), antifungal, antiviral and antimalarial drugs, so standard treatments become ineffective and infections persist, increasing the risk. of dissemination to others. The evolution of resistant strains is a natural phenomenon that occurs when microorganisms erroneously replicate themselves or when resistant traits are exchanged between them. The use and abuse of antimicrobial drugs accelerates the emergence of drug-resistant strains. Inadequate infection control practices, inadequate sanitation conditions and inappropriate food handling encourage the spread of antimicrobial resistance.
[200] In one embodiment of the second aspect of the present invention, the microorganism is an antibiotic-resistant microorganism selected from the group consisting of a gram positive bacterium, a gram negative bacterium, a biofilm forming bacterium, Streptococcus pneumoniae, Campylobacter , Neisseria gonorrhoeae, Salmonella (including drug-resistant non-typhoid Salmonella and drug-resistant Salmonella typhi serotype), Methicillin-resistant Staphylococcus aureus (MRSA), Shigella, Vancomycin-resistant Enterococcus (VRE), Vancomycin-resistant Staphylococcus aureus (VRSA) , Erythromycin-resistant Group A Streptococcus Clindamycin-resistant Group B Streptococcus, Carbapenem-resistant Enterobacteriaceae (CRE), drug-resistant tuberculosis, extended-spectrum Enterobacteriaceae (ESBL), multidrug-resistant Acinetobacter (including MRAB), Clostridium difficile, E Enteropathogenic coli (EPEC), Pseudomonas aeruginosa, H. pylori, Streptococcus a nginosus and Uropathogenic E. coli (UPEC).
[201] However, the practice of the second aspect of the invention is not limited to the treatment or prophylaxis of resistant microorganisms. The second aspect of the present invention may also be used to increase the sensitivity of non-resistant microorganisms to antimicrobial agents and thus provide a treatment that utilizes lower dosages of antimicrobial agents and/or shorter treatment durations with antimicrobial agents and/or results of more effective treatment with antimicrobial agents.
[202] Therefore, in a further embodiment of the second aspect of the present invention, the method, or the product for use, is to potentiate the antimicrobial (including antibiotic) effect of the agent administered separately, simultaneously or sequentially one or more antimicrobial agents ( including one or more antibiotics). For example, in a further embodiment, the amount of one or more antimicrobial agents administered separately, simultaneously or sequentially (including one or more antibiotics) may be less than a therapeutically effective or therapeutically ideal dose of one or more antimicrobial agents (including one or more antibiotics). plus antibiotics) when administered to the patient or animal not receiving the product. In another embodiment, the amount of one or more antimicrobial agents administered separately, simultaneously or sequentially (including one or more antibiotics) may be 5%, 10%, 15%, 20%, 30%, 40%, 50% 60 %, 70%, 80% or more, less than a therapeutically effective or therapeutically ideal dose of the one or more antibiotics when administered to the patient or animal not receiving the product. In another modality, the treatment duration of the patient receiving the second modality treatment or prophylaxis may be 5%, 10%, 15%, 20%, 30%, 40%, 50%, 60%, 70%, 80% or more, less than the required treatment duration when the patient or animal does not receive the product.
[203] Preferably, the subject to be treated in accordance with any embodiment of the second aspect of the present invention is a human patient. The human being can be a male. Alternatively, the human may be a female. A human being can be up to or older than 1 month, 2 months, 3 months, 4 months, 5 months, 6 months, 1 year, 2 years, 3 years, 4 years, 5 years, 6 years, 7 years, 8 years 9 years, 10 years, 15 years 20 years, 30 years, 40 years, 50 years, 60 years, 70 years, 80 years, 90 years, 100 years or more.
[204]Alternatively, the subject to be treated in accordance with any embodiment of the second aspect of the present invention may be an animal. Without limitation, animals for treatment or prophylaxis according to the second aspect of the present invention may be selected from the group consisting of domestic, wild and farmed animals, including mammals, marine animals, amphibians, birds, reptiles, insects and other invertebrates. . Without limitation, exemplary animals for treatment or prophylaxis include poultry, such as a chicken, turkey, geese, quail, pheasant, or duck; livestock, such as cattle, sheep, goat or swine, alpaca, banteng, bison, camel, cat, deer, dog, donkey, gayal, guinea pig, horse, llama, mule, rabbit, reindeer, water buffalo, yak; zoo animals; captive animals; hunting animals; marine or aquatic, such as fish (including freshwater and saltwater fish, farmed fish and ornamental fish) and crustaceans, including but not limited to oysters, mussels, clams, shrimp, lobsters, crayfish, crabs, cuttlefish, octopus and squid; domestic animals such as a cat or dog, a rodent (mouse, rats, guinea pigs, hamsters), horse.
[205] The one or more antimicrobial agents used and/or referred to in the second aspect of the present invention include those listed and discussed in Section III.B of this application, below. In one embodiment, at least one, or all, of the one or more antimicrobial agents is/are an antibiotic. The one or more antibiotics may, for example, be selected from the group consisting of aminoglycosides, ansakines, carbapenems, cephalosporins, glycopeptides, lincosamides, lipopeptides, macrolides, monobactams, nitrofurans, oxazolidinones, penicillins, polypeptides, quinolones/fluoroquinolones, sulfonamides, tetracyclines , clofazimine, dapsone, capreomycin, cycloserine, ethambutol, ethionamide, isoniazid, pyrazinamide, rifampicin (rifampicin), rifabutin, rifapentine, streptomycin, arsphenamine, chloramphenicol, fosfomycin, fusidic acid, metronidazole, mupirocin, platensimycin, quinupristin/dalfopristine, thiamphenicol, tigete , tinidazole and trimethoprim; and combinations thereof. More specific antibiotics suitable for use in accordance with the second aspect of the present invention include those listed and discussed in section III.B of this application below.
[206] In an embodiment of the second aspect of the present invention, the product comprising one or more compounds having the structure of Formula A or B, or other compounds of the invention as described further in section III.A of this application below is selected from the group consisting of a pharmaceutical or veterinary product. In one embodiment, it is a pharmaceutical product suitable for use with humans. In another embodiment, it is a veterinary product suitable for use with animals, including but not limited to one or more animals selected from the group consisting of domestic, wild and farmed animals, including mammals, marine animals, amphibians, birds, reptiles, insects and other invertebrates. Without limitation, exemplary animals for treatment or prophylaxis include poultry, such as a chicken, turkey, geese, quail, pheasant, or duck; livestock, such as cattle, sheep, goat or swine, alpaca, banteng, bison, camel, cat, deer, dog, donkey, gayal, guinea pig, horse, llama, mule, rabbit, reindeer, water buffalo, yak; zoo animals; captive animals; hunting animals; marine or aquatic, such as fish (including freshwater and saltwater fish, farmed fish and ornamental fish) and crustaceans, including but not limited to oysters, mussels, clams, shrimp, lobsters, crayfish, crabs, cuttlefish, octopus and squid; domestic animals such as a cat or dog, a rodent (mouse, rats, guinea pigs, hamsters), horse. In a preferred embodiment, the animal is a chicken, for example, a meat-like chicken, such as a broiler, or a laying hen, such as a pullet or a broiler or a broiler. The pharmaceutical or veterinary product may or may not additionally include one or more antimicrobial agents (in the embodiment it does not, then, in accordance with the second aspect of the present invention, the product and the microbial agent are intended to be administered to the subject in separately, simultaneously or sequentially). The pharmaceutical or veterinary product may include one or more excipients, as discussed in section III. C of this order, below. The pharmaceutical or veterinary product may be presented as a parenteral formulation, as discussed below in section III.C.1 of this application, including a controlled release formulation, as discussed below in section III.C.1(a) of this application. and injectable or implantable formulation, as discussed below in section III.C.1(b) of this application. The pharmaceutical or veterinary product may be presented as an enteral formulation, as discussed below in section III.C.2 of this application, including a controlled release enteral formulation, as discussed below in section III.C.2(a) of this application. application, with further reference to extended-release dosage forms and delayed-release dosage forms as discussed therein. The pharmaceutical or veterinary product may be presented as a topical formulation as discussed below in section III.C.3 of this application, including as an emulsion, lotion, cream, ointment, gel or foam as discussed in parts (a), (b) ), (c), (d) (e) and (f), respectively, in section III.C.3 of this order.
[207] In another embodiment, the product comprising one or more compounds having the structure of Formula A or B, or other compounds of the invention, as described further in Section III.A of this application below is a medical device. The device may or may not additionally include the one or more antimicrobial agents (in the embodiment it does not, then, in accordance with the second aspect of the present invention, the product and the microbial agent are intended to be administered to the subject in separate compositions, separately, simultaneously or sequentially). Medical devices that may comprise one or more compounds as defined in section III.A of this application may include, without limitation, wound dressings or medical implants. Other examples include tubes and other surface medical devices such as urinary catheter, stents, mucosal extraction catheter, suction catheter, umbilical cannula, contact lenses, intrauterine devices, intravaginal and intraintestinal devices, endotracheal tubes, bronchoscopes, prostheses dental and orthodontic devices, surgical instruments, dental instruments, tubing, dental water lines, dental drainage tubes, tissue, paper, indicator strips (e.g. paper indicator strips or plastic indicator strips), adhesives (e.g. adhesives hydrogels, hot adhesives or solvent-based adhesives), bandages, tissue dressings or healing devices and occlusive patches and any other surface devices used in the medical field. Devices can include electrodes, external prostheses, fixation tapes, compression bandages and monitors of various types. Medical devices also include any device that can be placed at the site of insertion or implantation, such as the skin near the site of insertion or implantation, and that includes at least one surface that is susceptible to colonization by microorganisms embedded in biofilm. In a specific embodiment, a composition is integrated into an adhesive, such as a tape, thereby providing an adhesive, which can present and/or distribute one or more compounds on at least one surface of the adhesive. In a particularly preferred embodiment, the following devices may comprise, include, and/or be coated with the compounds: catheters, including central venous catheters, urinary catheters, dialysis catheters, and indwelling catheters (e.g., catheters for hemodialysis and for administering agents chemotherapy drugs), cardiac implants, including mechanical heart valves, stents, ventricular assist devices, pacemakers, cardiac rhythm management devices (CRM), cardiac resynchronization therapy (CRT) devices, and implantable cardioverter defibrillators (ICDs), grafts synthetic vascular shunts, arteriovascular shunts, cerebrospinal fluid shunts, cochlear devices, prosthetic joints, orthopedic implants, internal fixation devices, bone cements, percutaneous sutures, surgical mesh and surgical corrections including hernia repair meshes and adhesives, reconstruction meshes of breast and adhesive breast and face lift meshes and adhesives, slings and meshes and adhesives for pelvic floor reconstruction, tracheal and ventilation tubes, dressings, biological implants (including allografts, xenografts and autografts), penile implants, intrauterine devices , endotracheal tubes, and contact lenses.
[208] In another embodiment, the product comprising one or more compounds having the structure of Formula A or B, or other compounds of the invention, as described below in section III.A of this application below is a dietary product. The dietary product may or may not additionally include one or more antimicrobial agents (in the embodiment it does not, then, in accordance with the second aspect of the present invention, the product and the microbial agent are intended to be administered to the subject in separate compositions, separately, simultaneously or sequentially). Dietary products may include, for example, food products, dietary supplements, beverages and any other orally taken compositions that incorporate one or more compounds having the structure of Formula A or B or other compounds of the invention as described in section III. .A of this order below.
[209] The one or more compounds are selected from the group consisting of a complex of an amino acid with Fe III, and a complex of an a-hydroxy acid with Fe III, or salts and/or hydrates thereof. The one or more compounds may or may not be selected from any one or more of the group consisting of a complex of quinic acid with Fe III (such as a complex with the structure of Formula IX), a complex of L-tyrosine with Fe III (such as a complex having the structure of Formula VIII), a complex of L-DOPA with Fe III (such as a complex having the structure of Formula VII) and a complex of L-phenylalanine with Fe III. Accordingly, in one embodiment, a complex of L-tyrosine with Fe III (such as a complex with the structure of Formula VIII) is particularly preferred. Optionally, one or more compounds is not a complex of quinic acid with Fe III (such as a complex having the structure of Formula IX).
[210] A complex of quinic acid with Fe III (Fe-QA, also called FeQ), as defined by Formula IX, can be used with any one or more of the above antibiotics or other antimicrobials, either formulated together in the same composition. for administration or presented in separate compositions for use separately, simultaneously or sequentially.
[211] A complex of quinic acid with Fe III (Fe-QA, also called FeQ), as defined by Formula IX, can be used with any one or more of the foregoing antibiotics or antimicrobials, either formulated together in the same composition to administration or presented in separate compositions for use separately, simultaneously or sequentially.
[212] A complex of L-tyrosine with Fe III (Fe-Tyr), as defined by Formula VIII, can be used with any one or more of the above antibiotics or antimicrobials, either formulated together in the same composition for administration or presented in compositions for use separately, simultaneously or sequentially.
[213] A complex of L-DOPA with Fe III (3,4-dihydrophenylalanine) (Fe-DOPA), as defined by Formula VII, can be with any one or more of the above antibiotics or antibiotics, formulated together in the same composition for administration or presented in separate compositions for use separately, simultaneously or sequentially.
[214] A complex of L-phenylalanine with Fe III (Fe-Phe) can be used with any one or more of the above antibiotics or antibiotics, formulated together in the same composition for administration or presented in separate compositions for use separately, simultaneously. or sequentially.
[215] In one embodiment, wherein the product is selected from the group consisting of a pharmaceutical or veterinary product, a medical device or a dietary product comprising the combination of one or more compounds having the structure having the structure of Formula A or B, or other compounds as described further in section III.A of this application below and one or more antibiotics or other antimicrobial agents, then optionally the amount (by weight or in moles) and/or concentration of the one or more antibiotics and/or other agents antimicrobials in the product is less than (e.g. a reduction of up to, or at least 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10%, 15 %, 20%, 35%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or greater) compared to a therapeutically effective or therapeutically optimal amount or concentration of one or more antibiotics and/or other antimicrobial agents when administered to the patient or animal not receiving the product. to.
[216] The product may be presented in a unit dosage formulation and, optionally, the unit dosage formulation may include one or more antibiotics and/or other antimicrobial agents in an amount (by weight or in moles) and/or concentration that is less than (e.g. a reduction of up to or at least 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10%, 15%, 20 %, 35% 30%, 40%, 50%, 60%, 70%, 80%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99 % or more) compared to a therapeutically effective or therapeutically optimal dose of one or more antibiotics when administered to the patient or animal not receiving the product.
[217] There is also provided a product per se, such as a pharmaceutical or veterinary product, a medical device or a dietary product, which is suitable for use in accordance with the above methods and uses of the second aspect of the present invention. The product comprises one or more compounds having the structure of Formula A or B, or other compounds of the invention as described further in section III.A of this application below, optionally in combination with one or more antibiotics or other antimicrobial agents as discussed herein in with respect to the second aspect of the present invention. As discussed above, in the option where the product comprises one or more antibiotics or other antimicrobial agents, then they may be included in an amount, concentration, and/or with a release profile that is typically subtherapeutic or sub-therapeutic for treatment or prophylaxis of a microbial infection or colonization.
[218] Also provided herein is a method for sensitizing and/or reducing the tolerance of one or more microorganisms to a selected antimicrobial agent, the method comprising exposing the one or more microorganisms to one or more compounds with the structure of Formula A or B, or other compounds of the invention as described further in section III.A of this application below. In other words, the second aspect of the present invention also provides the use of one or more compounds having the structure of Formula A or B, or other compounds of the invention as described further in Section III.A of this application below to increase the sensitivity and /or reduce the tolerance of one or more microorganisms to a selected antimicrobial agent. The microorganisms may or may not be microorganisms that are resistant to the selected antimicrobial agent. Thus, in one option, the in vivo and/or in vitro growth of one or more microorganisms may normally be unaffected by the selected antimicrobial agent (any one or at a selected concentration) in the absence of one or more compounds with the structure of Formula A or B, or other compounds of the invention as described further in section III.A of this application below, while exposure to one or more compounds may cause the in vivo and/or in vitro growth of the one or more microorganisms to be reduced (e.g. 1%, 2%, 3%, 4%, 5%, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 95 %, 99%, or substantially 100%) by exposure to the selected antimicrobial agent (any, or the selected concentration). In this context, a "selected concentration" includes concentrations that are pharmaceutically and medically acceptable for use with patients and/or animals and lower concentrations, such as 1%, 2%, 3%, 4%, 5%, 10%, 20 %, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 95% or 99% lower, which may or may not be subtherapeutic. In another option, it may already be possible to reduce the in vivo and/or in vitro growth of the one or more microorganisms by exposure to the antimicrobial agent (any one, or at a selected concentration) in the absence of one or more compounds with the structure of Formula A or B, or other compounds of the invention as described further in section III.A of this application below, while exposure to one or more compounds may cause an increase in sensitivity and/or a reduction in tolerance to the antimicrobial agent of such that the in vivo and/or in vitro growth of the one or more microorganisms is further reduced (e.g. 1%, 2%, 3%, 4%, 5%, 10%, 20%, 30%, 40% , 50%, 60%, 70%, 80%, 90%, 95%, 99% or substantially 100%) by the antimicrobial agent and/or an equivalent level of growth reduction may be achieved with a lower concentration or amount of the antimicrobial agent (e.g. using an amount (by weight) or concentration that is reduced by 1% 2%, 3%, 4%, 5%, 10%, 20% , 30%, 40%, 50%, 60%, 70%, 80%, 90%, 95%, 99% or substantially 100%) and/or the treatment period can be shortened, for example by 1, 2, 3, 4, 5, 6, 7 or more days.
[219] A further embodiment provides a method for preparing a product per se, such as a pharmaceutical or veterinary product, a medical device, or a dietary product, which is suitable for use in accordance with the methods and uses above of the second aspect. of the present invention. The method may include the step of mixing the one or more compounds having the structure of Formula A or B or other compounds of the invention as described further in section III.A of this application below without one or more additional product components, and so on. forming the product. The method may include forming the product (optionally without one or more compounds) and then spraying or otherwise applying one or more compounds having the structure of Formula A or B, or other compounds of the invention as described later in section III. A of this order below, for the product. The method may include forming the product (optionally without one or more compounds) and then coating the product with one or more compounds having the structure of Formula A or B, or other compounds of the invention as described further in section III. . That of this application below, for example, as described further in the context of coatings.
[220] One or more compounds having the structure of Formula A or B, or other compounds as described further in section III.A of this application below ("Component 1") may be administered simultaneously, separately or sequentially with one or more antibiotics and/or other antimicrobial agents ("Component 2").
[221] In the context of simultaneous administration, Components 1 and 2 may be present in the same product for administration to the patient or animal. Alternatively, Components 1 and 2 may be present in separate products that are administered at the same time, although this may be via different routes. For example, both Components 1 and 2 can be administered, in separate products but at the same time, via an enteral route. In another embodiment, Component 1 can be administered enterally and Component 2 can be administered at the same time by a parenteral route. In another embodiment, Component 1 can be administered parenterally and Component 2 can be administered at the same time by an enteral route. In another embodiment, both Components 1 and 2 can be administered, as separate products, but at the same time, via a parenteral route.
[222]In the context of separate and/or sequential administration, Components 1 and 2 are administered to the patient at different times. Component 1 can be administered before Component 2, or Component 2 can be administered before Component 1. Preferably, the time period between administration of Components 1 and 2 is less than the time it takes the subject to clear a effective amount of the component administered for the first time, such that effective amounts of Components 1 and 2 will be present in the subject simultaneously. However, this may not be essential. In either case, the time it takes the subject to clear an effective amount of the component administered for the first time will vary depending on the nature of the component, the route of administration and the form administered which may, for example, be a device, product or formulation. slow, delayed or extended release. Administration of Components 1 and 2 (in any order) may be temporarily separated by up to, about, or at least 10 seconds, 20 seconds, 30 seconds, 40 seconds, 50 seconds, 1 minute, 5 minutes 10 minutes, 20 Minutes, 30 minutes, 40 minutes, 50 minutes, 1 hour, 2 hours, 3 hours, 4 hours, 5 hours, 6 hours, 7 hours, 8 hours, 9 hours, 10 hours, 12 hours, 14 hours, 16 hours, 18 hours, 20 hours, 22 hours 1 day, 2 days, 3 days, 4 days, 5 days, 6 days, 1 week, 2 weeks, 3 weeks, 1 month or more. Sequential administration includes the meaning of repeated and alternating administrations of Components 1 and 2 (in any order), wherein administration of one or both components may be repeated any number of times, such as twice, three times, four times, five times, 10 times, 20 times, 30 times or more.
[223]Repeated administration of one or both components, whether administered simultaneously, separately, or sequentially, may occur as often as is therapeutically necessary, and may include continuous administration (e.g., by intravenous infusion), administration up to, approximately, or at least 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 24 or 24 hours, 1, 2, 3, 4, 5, 6, 7 days, or every 1, 2, 3, 4 or more weeks, throughout the treatment period.
[224] The period of treatment in accordance with the second aspect of the present invention is typically selected to achieve an effective therapeutic or prophylactic result and will be evaluated accordingly by the skilled practitioner. Examples of suitable periods for treatment may include 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13 or 14 days, about 1, 2, 3, or 4 weeks , or longer. C. Inhibition of the formation and treatment of preformed biofilms
[225] A third aspect of the present invention is based on the surprising finding that compounds having the structure of Formula A or B, or other compounds of the invention of the present invention, as described further in Section III.A of this application, have a wide range of action in treating and dispersing pre-existing biofilms and inhibiting the development of biofilms created by a wide variety of bacteria and other microbial sources, and that this action is effective in a variety of environments.
[226] Accordingly, a third aspect of the present invention provides a method of inhibiting biofilm accumulation and/or disrupting a pre-existing biofilm, in or on a subject or article in need thereof, the method comprising administering to the subject or article an effective amount of one or more compounds having the structure of Formula A or B, or other compounds of the invention of the present invention as described further in Section III.A of this application.
[227] In other words, the third aspect of the present invention provides the use of one or more compounds having the structure of Formula A or B, or other compounds of the invention of the present invention as described further in section III.A of this application. to inhibit biofilm accumulation and/or disrupt a pre-existing biofilm, in or on a subject or article in need thereof.
[228] In one embodiment, the one or more compounds having the structure of Formula A or a salt and/or hydrate thereof, or a functional variant thereof, for use in accordance with the third aspect of the present invention are selected from the group which consists of a complex of an amino acid or an a-hydroxy acid with Fe III, such as a complex of quinic acid with Fe III, a complex of L-tyrosine with Fe III, a complex of L-DOPA with Fe III, a complex of L-Phenylalanine with Fe III, the compounds represented by Formula I, Formula II, Formula III, Formula IV, Formula V, Formula VI, Formula VII, Formula VIII, Formula IX, Formula X, Formula XI, Formula XII, Formula XIII , Formulas XIV, a compound selected from the group consisting of a compound that binds to major outer membrane proteins (MOMPs) or Campylobacter FlaA, a synthetic human histo-blood group antigen, a histo-blood group antigen mimetic human or a synthetic sugar. Particularly preferred compounds may or may not include Fe-QA, Fe-Tyr and/or Fe-DOPA. 1. Organisms to be treated, inhibited or killed
[229] "Biofilm" as used herein refers to any group of microorganisms in which cells adhere to one another on a surface.
[230]The formation of a biofilm begins with the attachment of free-floating microorganisms to a surface. These early settlers adhere to the surface initially through weak adhesion and reversible through van der Waals forces. If colonizers are not immediately separated from the surface, they can anchor themselves more permanently using cell adhesion structures such as pili. Some species are not able to attach to a surface on their own, but are sometimes able to anchor to the matrix or directly to previous colonists. It is during this colonization that cells are able to communicate via quorum sensing. Once colonization begins, the biofilm grows through a combination of cell division and recruitment. Polysaccharide matrices typically contain bacterial biofilms. The final stage of biofilm formation is known as dispersion, and it is the stage where the biofilm is established and can only change in shape and size.
[231] In one embodiment, a biofilm may comprise, essentially consist of, or consist of, microbial cells growing in a biofilm that are physiologically distinct from planktonic cells of the same organism, which, in contrast, are individual cells. Optionally, a biofilm may comprise, consist essentially of, or consist of, a bacterial cell species or strain.
[232] In an alternative, a biofilm may comprise, essentially consist of, or consist of, more than one species or strain of the bacterial cell, such as up to at least 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 30, 40, 50, 60 70, 80, 90, 100, 150, 200, 250, 300, 350, 400, 450, 500, 600, 700, 800, 900, 1000 or more different species or strains of bacterial cells.
[233] Bacterial species or strains in biofilms may include bacteria selected from one or more gram negative, gram positive, aerobic and anaerobic and/or archaeal bacteria.
[234] Accordingly, compositions and methods for inhibiting, reducing or removing biofilm-forming bacteria and bacterial infections are provided by the third aspect of the present invention.
[235] In accordance with some embodiments of the third aspect of the present invention, the biofilm-forming bacteria to be inhibited, reduced, removed or treated may be gram-negative and/or gram-positive bacteria, such as Pseudomonas aeruginosa, Campylobacter jejuni, Helicobacter pylori, Escherichia coli, Enteropathogenic Escherichia coli (EPEC), Uropathogenic Escherichia coli (UPEC), Staphylococcus epidermidis, Staphylococcus aureus, and Enterococcus faecalis.
[236] The following are representative organisms that may be killed or inhibited by growth, or their ability to produce or maintain biofilms degraded, reduced, inhibited or prevented in accordance with the third aspect of the present invention.
[237] A form of biofilm of particular interest in certain embodiments of the third aspect of the present invention is the biofilm that forms dental plaque. The effectiveness of the present invention against dental plaque is demonstrated in Example 17. The biofilm on dental plaque typically comprises a variety of microbial organisms, including aerobic and anaerobic bacteria, and typically includes over 700 different species of bacteria and archaea. Dental plaque biofilms are responsible for many diseases common to the oral cavity, including less common dental caries, periodontitis, gingivitis and peri-implantitis (similar to periodontitis but with dental implants), but biofilms can also be present on healthy teeth.
[238]Suitably, the third aspect of the present invention also provides methods and uses for preventing or inhibiting the formation of, for treating, or for reversing or removing conditions including dental plaque, dental caries, periodontitis, gingivitis and the less common peri-implantitis. . Said method or use may comprise administering a composition according to the third aspect of the present invention to the mouth of a subject, to thereby achieve the intended effect.
[239]In the context of dental products, and in the context of inhibition and/or removal of dental plaque, a concentration of 340 μM is demonstrated in Example 17, although higher or lower concentrations of the one or more compounds according to section III.A below may also be suitable. For example, dental products may present the oral cavity or teeth with one or more of said compounds at a concentration within the range of about 1μM to about 1M, such as about or up to 10 μM, 20μM, 30μM, 40 μm, 50 μm, 60 μm, 70 μm, 80 μm, 90 μm, 100 μm, 110 μm, 120μm, 130μm, 140 μm, 150 μm, 160 μm, 170 μm, 180 μm, 190 μm, 200 μm, 210 μm , 220μm, 230μm, 240 μm, 250 μm, 260 μm, 270 μm, 280 μm, 290 μm, 300 μm, 310 μm, 320μm, 330μm, 340 μm, 350 μm, 360 μm, 370 μm, 380 μm, 390 μm , 400 μm, 420μm, 430μm, 440 μm, 450 μm, 460 μm, 470 μm, 480 μm, 490 μm, 490 μm, 500 μm, 510 μm, 520μm, 530μm, 540 μm, 550 μm, 560 μm, 570 μm , 580 μm, 60 μm, 610 μm, 620μm, 630μm, 640 μm, 650 μm, 660 μm, 60 μm, 680 μm, 680 μm, 690 μm, 700 μm, 710 μm, 720μm, 730μm, 740 μm, 750 μm , 760 μm, 780 μm, 780 μm, 790 μm, 800 μm, 810 μm, 820μm, 830μm, 840 μm, 850 μm, 860 μm, 870 μm, 880 μm, 880 μm, 890 μm, 900 μm, 910 μm, 920μm , 940 µM, 950 µM, 960 µM, 970 µM, 980 µM, 990 µM, 1 mM, 2 mM, 3 mM, 4 mM, 5mM, 6mM, 7mM, 8mM, 9mM, 10mM, 15mM, 20mM 25mM, 30mM, 35mM, 40mM, 45mM, 50mM, 60mM, 70mM, 80mM, 90mM or more. Optionally, the concentration may be: (a) up to 1 μM, 2μM, 3μM, 4μM, 5μM, 10μM, 15 μM, 20 μM, 25 μM, 30 μM; (b) within a selected group range consisting of 35 to 335 μM, 40 to 300 μM, 50 to 300 μM, 50 to 250 μM, 50 to 200 μM, 60 to 300 μM, 60 to 250 μM, 60 to 200 µM, 80 to 300 µM, 80 to 250 µM, 80 to 200 µM, 100 to 300 µM, 100 to 250 µM, or 100 to 200 µM; or (c) at least, or about, 345 µM, 350 µM, 360 µM, 370 µM, 380 µM, 390 µM, 400 µM, 450 µM, 0.5 mM, 1 mM, 2 mM or more.
[240] Optionally, the concentration may be within a range selected from the group consisting of about 1 μM to about 1 mM, or about 30 μM to about 0.5 mM, or about 60 μM to about 0.5 mM. 0.4 mM.
[241] Another form of biofilm of particular interest to the third aspect of the present invention is the biofilm on medical devices, including contact lenses. Biofilms on contact lenses may, for example, comprise, consist essentially of, or consist of one or more bacteria selected from Archromobacter, Delftia, Staphylococcus, Stenotrophomonas, and Streptococci species, and Pseudomonas aeruginosa.
[242] Another form of biofilm of particular interest in the present invention are biofilms formed on the skin, for example biofilms that comprise, consist essentially of, or consist of Propionibacterium acnes. Suitably, the third aspect of the present invention also provides methods and uses for preventing or inhibiting the formation of, for treating, or for reversing or removing acne and other microbially induced skin conditions, including recalcitrant and/or antibiotic resistant conditions, the A method or use which comprises typically administering a composition according to the third aspect of the present invention to the skin of a subject to thereby achieve the intended effect.
[243] Another form of biofilm that may or may not be of particular interest in the third aspect of the present invention are biofilms comprising, essentially consisting of, or consisting of, a class of epsilon proteobacteria, such as the spirilloid Wolinella spp. , Helicobacter spp., and more particularly Campylobacter spp. In one embodiment, application of the third aspect of the present invention to biofilms that comprise, consist essentially of, or consist of Campylobacter spp. may, or otherwise may not, be of interest. Many other types of biofilms are of interest to the present invention, other examples of which are discussed in further sections of this application.
[244] Campylobacter are gram-negative, coiled rod-shaped bacteria with a single flagellum at one or both poles. They belong to the epsilon proteobacteria class and are closely related to Helicobacter and Wolinella. At least a dozen Campylobacter species have been implicated in human disease, with C. jejuni and C. coli being the least common.
[245] Campylobacter jejuni is the leading cause of human bacterial gastroenteritis (Pearson, et al., Appl Environ Microbiol., 59:987-996 (1993)). The four main sources of infection are raw meat (particularly poultry), untreated water, raw milk, and pets (Humphrey et al. J Appl Bacteriol. 61: 125-132 (1986) and Skirrow, Int J Microbiol. Alimentar., 12:9-16 (1991)). It has also been suggested that, while not universally the case (Humphrey et al., Public Health Lab Serv Microbiol Digest., 13:86-88. 91996), Jacobs-Reitsma, et al., Epidemiol Infect., 114:413-421 (1995), and Lindblom, et al., J Hyg., 96: 385-391 (1986)), survival in the water systems of animal husbandry facilities and animal processing units promotes infection in animals and contamination. crossbreeding of animal carcasses (Humphrey et al. Epidemiol Infect., 98:263-269 (1987), Kazwala, et al., Vet Rec. 1990; 126:305-306. (1990) and Pearson, et al., Appl Environ Microbiol., 59:987-996 (1993)). Thus, the survival of C. jejuni in aquatic environments is important both directly and indirectly in the cause of human disease.
[246] Campylobacter spp. have outer membrane proteins (OMPs) (Buchanan, Curr. Opin. Struc. Biol.,9 (40:455461 (1999); Huyer, et al., FEMS Microbiol. Lett., 37(3):247-250 ( 1986)] Major outer membrane proteins (MOMPs) have unique structural features, and function as porins that are useful for binding bacteria and their environment. Campylobacter spp. have polar flagella that provide the necessary motility for intestinal colonization. Campylobacter flagellin gene have two similar copies: flaA and flaB. The length of the coding regions for the flaA and flaB sequences is about 1.7 kilobases, and the flaA and flaB sequences are located about 180 bases from each other (Meinersmann, et al., Microbiology, 146(9):2283 (2000 )).
[247] In an embodiment of the third aspect of the present invention, the disclosed compositions bind to major outer membrane proteins (MOMPs) or Campylobacter FlaA and prevent the bound MOMPs and bound FlaA from binding or associating with their ligands in :other Campylobacter bacteria; other species of bacteria; biofilm or biofilm components; or to surfaces. Through binding to MOMPs and FlaA, the compounds inhibit bacteria from attaching to surfaces or each other to produce biofilm. Inhibition of binding can be achieved by interfering with the binding of natural ligands of MOMPs or FlaA or by physically inhibiting the association of bacteria expressing MOMPs or FlaA with other organisms or surfaces.
[248] In another embodiment of the third aspect of the present invention, the disclosed compositions also bind the Campylobacter MOMP protein when MOMP has been mutated to prevent O-glycosylation by mutating Thr-268 to glycine to form MOMP-T (also referred to as such as MOMPT268G). As shown in Table 1, the expression of the MOMPT268G protein was found to increase 10-fold compared to wild type. Treatment of the MOMPT268G strain with the compositions does not affect planktonic growth, but partially inhibits biofilm formation, demonstrating that the compositions bind non-glycosylated MOMP with lower affinity.
[249] As shown in Table 1, the expression of the MOMPT268G protein was found to be increased 10-fold compared to wild type. Regardless of whether or not the MOMP is glycosylated, the compositions disclosed herein are still effective against mixed populations of glycosylated and non-glycosylated Campylobacter. In a mixed population of glycosylated and non-glycosylated forms, the wild-type glycosylated form of Campylobacter largely outnumbers the mutant non-glycosylated form, and over time the non-glycosylated bacteria disappear and the glycosylated bacteria become the only bacteria present.
[250] Biofilms are commonly found on solid substrates submerged or exposed to an aqueous solution, although they can form as floating mats over liquid surfaces. Biofilms can form on a multitude of surfaces. For example, biofilms can grow in showers very easily, as they provide a moist, warm environment for the biofilm to thrive. Biofilms can form on the inside of water and sewer pipes and cause clogging and corrosion. Biofilms on floors and counters can hamper sanitation in food preparation areas. Biofilms can form in cooling or heating systems and are known to reduce heat transfer in these systems.
[251] A method, or use, according to a third aspect of the present invention includes administering an effective amount of one or more compounds as defined in section III.A of this application to a subject in need thereof, to inhibit formations of biofilm, or alternatively, to reduce and/or remove biofilm formation. The one or more compounds may be administered alone, or in combination with an antimicrobial agent, such as an antibiotic.
[252] In certain embodiments of the third aspect of the present invention, in the context of treating subjects (such as humans or animals) it may be desirable to provide continuous delivery of one or more compounds to a subject in need thereof. For intravenous or intra-arterial routes, this may be accomplished using drip systems, such as by intravenous administration. For topical applications, repeated application may be made or a patch may be used to provide continuous administration of the compounds over an extended period. For example, the compounds can be administered to a chronic wound from a wound dressing. The dressing can also contain one or more antibiotics, and if necessary, the wound dressing can be changed frequently. Compounds may also be provided in a conjugated form (eg, as shown in Figures 15A-C and Figures 16A and B) so that they are immobilized on a surface.
[253] In other embodiments of the third aspect of the present invention, the method includes contacting a surface with an effective amount of the compounds to inhibit biofilm buildup, reduce buildup biofilm, and/or remove buildup biofilm. "Contact" includes, but is not limited to, touching, impregnating, compounding, mixing, integrating, coating, spraying, dipping, washing, irrigating and cleaning. In certain embodiments, it may be desirable to provide continuous delivery of one or more compounds to the surface or system to be treated. The compositions can be used to coat, impregnate, wash or rinse a tube surface or a medical device, especially an insertable medical device. Tubes include, but are not limited to, disposable, indwelling and indwelling catheters, long-term urinary devices, tissue-connecting urinary devices, wound drainage tubes, ventricular catheters, endotracheal tubes, breathing tubes, feeding tubes, lines dairies, oil and gas pipelines and potable water lines. When an object is piping (e.g. dental unit water line, dairy line, food and beverage processing line, etc.), a composition may be poured into the piping and both ends of the piping tightened such that the composition is retained within the lumen of the tubing. The tubing is then allowed to fill with the composition for a period sufficient to remove substantially all microorganisms from at least one surface of the object, generally for at least about 1 minute to about 48 hours. Alternatively, the tube may be washed by pouring a composition into the lumen of the tube for an amount of time sufficient to prevent substantial growth of all microorganisms incorporated into the biofilm. Such flushing may be required only once, or it may be required at regular intervals throughout the life of the pipeline's use. The concentrations of active components in a composition can be varied as desired or necessary to decrease the amount of time the composition is in contact with a medical device.
[254] The methods allow disinfection, inhibition or prevention of biofilm formation on surfaces to be treated and reduction of the transmission of biofilms that form microorganisms from the surface to another surface. The number of bacterial colony forming units (cfu) on the surface to be treated with the compounds can be reduced by 50%, by 60%, by 70%, by 80%, by 90% or by 100% or the accumulation of units of bacterial colony formation on the treated surface can be reduced by 50%, 60%, 70%, 80%, 90% or 100%.
[255] In an embodiment of the third aspect of the present invention, compositions and articles, including, but not limited to, pharmaceutical and veterinary compositions, food or animal feed additive compositions, and dental products including chewers can be prepared from of one or more compounds as defined above, optionally formulated and/or used in combination with one or more antibiotics or other antimicrobial agents and these compositions may further be used for the treatment or prophylaxis of a microbial infection or biofilm formed by bacteria or other microbes -organisms including one or more of the following: S. epidermidis, E. faecalis, E. coli, S. aureus including Vancomycin-resistant Staphylococcus aureus (VRSA) and Methicillin-resistant Staphylococcus aureus (MRSA), Enteropathogenic Escherichia coli (EPEC) , Uropathogenic Escherichia coli (UPEC), Pseudomonas, Streptococcus pneumoniae, Streptococcus anginosus, Neisseria gonor rhoeae, Salmonella (including drug-resistant non-typhoid Salmonella, drug-resistant serotype typhi Salmonella, Salmonella Enteritidis, Salmonella Typhimurium, Mycoplasma, Eimeria, Enterococci, Shigella, Vancomycin-resistant Enterococcus (VRE), Erythromycin-resistant Group A Streptococcus Group Streptococcus Clindamycin-resistant B, Carbapenem-resistant Enterobacteriaceae (CRE), drug-resistant tuberculosis, extended-spectrum Enterobacteriaceae (ESBL), multidrug-resistant Acinetobacter (including MRAB), Clostridium difficile, Enteropathogenic E. coli (EPEC), Pseudomonas aeruginosa, Brachyspira , Propionibacterium acnes, and Clostridium perfringen. 2. Administration Methods
[256] In one embodiment, compounds and formulations, derivatives thereof, and combinations thereof for use in accordance with the third aspect of the present invention can be administered topically to a subject in need thereof in an amount effective to prevent or treat a condition. microbial infection, inhibiting biofilm accumulation or to reduce and/or remove accumulated biofilm.
[257] Any suitable topical formulation may be used, for example as described in Section III.C.3 of the present application, below, including emulsions (such as those described in Section III.C.3(a)), lotions (such as those described in section III.C.3(b)), creams (such as those described in section III.C.3(c)), ointments (such as those described in section III.C.3(c) d)), gels (such as those described in section III.C.3(e)), or foams (such as those described in section III.C.3(f)).
[258] The compositions may be used alone or in combination with known antimicrobial agents, such as those described later in section III.B of this application. As such, the compositions described with respect to the second aspect of the present invention may also be useful in the practice of the third aspect of the present invention.
[259] The compositions are useful for the treatment of topical conditions caused by the accumulation of biofilm by microorganisms including, but not limited to, gram-negative and gram-positive bacteria, including Staphylococcus (including but not limited to S. aureus and Staphylococcus epidermidis), Pseudomonas, E. coli., Streptococcus pyogenes (Reviewed in Nusbaum, et al., Skin Therapy Lett., 17(7):1-5 (2012 )), Propionibacterium acnes and Streptococcus anginosus.
[260] In some embodiments the compositions are used as a topical antibacterial medication for skin infections caused by Methicillin-resistant Staphylococcus aureus. Methicillin-resistant Staphylococcus aureus (MRSA) is a bacterium that is resistant to many antibiotics. The spectrum of disease caused by MRSA appears to be similar to that of Staphylococcus aureus in the community. Soft tissue infections (SSTIs), specifically boils (abscessible hair follicles or "boils"), carbuncles (coalesced masses of boils), and abscesses, are the most frequently reported clinical manifestations.
[261] The most common manifestations of community-associated MRSA are simple skin infections such as impetigo, boils, abscesses, folliculitis, and cellulitis. Others include children with minor skin infections (such as impetigo) and secondarily infected skin lesions (such as eczema, ulcers, or lacerations). The compositions can also be used to treat MRSA infections of the CNS, which include, but are not limited to, meningitis, brain abscess, subdural empyema, spinal epidural abscess. Reviewed in Liu et al., Clin Infect Dis., 52(3):e18-55 (2011 ).
[262] Additional examples of conditions that can be treated include atopic dermatitis, acne, bullous and non-bullous impetigo, pemphigus foliaceus, miliaria, boils (also known as boils), and chronic wounds such as diabetic foot ulcers, venous insufficiency ulcers. and pressure ulcers.
[263]In the context of treating acne, an effective concentration of 340 μM is demonstrated in Example 24, although higher or lower concentrations of the one or more compounds according to section III.A below may also be suitable for the treatment. acne and any of the other skin conditions as discussed here. For example, the treatment of these skin conditions in accordance with the present invention may utilize one or more of said compounds at a concentration within the range of about 1μM to about 1M, such as about or up to 10 μM, 20μM, 30μm, 40 μm, 50 μm, 70 μm, 80 μm, 90 μm, 100 μm, 110 μm, 120μm, 130μm, 140 μm, 150 μm, 160 μm, 170 μm, 180 μm, 190 μm, 200 μm , 210 μm, 220μm, 230μm, 240 μm, 250 μm, 260 μm, 270 μm, 280 μm, 280 μm, 290 μm, 300 μm, 310 μm, 320μm, 330μm, 340 μm, 350 μm, 360 μm, 370 μm, 380 μm , 390 μm, 400 μm, 410 μm, 420μm, 440 μm, 450 μm, 450 μm, 460 μm, 470 μm, 480 μm, 490 μm, 490 μm, 500 μm, 510 μm, 520μm, 530μm, 540 μm, 550 μm, 560 μm , 570 μm, 580 μm, 600 μm, 610 μm, 620μm, 630μm, 640 μm, 650 μm, 660 μm, 670 μm, 680 μm, 690 μm, 700 μm, 710 μm, 720μm, 730 μm, 740 μm , 750 μm, 760 μm, 780 μm, 780 μm, 790 μm, 800 μm, 810 μm, 820μm, 830μm, 840 μm, 850 μm, 860 μm, 870 μm, 880 μm, 880 μm, 890 μm, 900 μm, 910 μm, 920μM, 930μM, 940 μM, 950 μM, 960 μM, 970 μM, 980 μM, 990 μM, 1 mM, 2mM, 3mM, 4mM, 5mM, 6mM, 7mM, 8mM, 9mM, 10mM, 15mM, 20mM 25mM, 30mM, 35mM, 40mM, 45mM, 50mM, 60mM, 70mM, 80mM, 90mM, 100mM, 150mM, 200mM, 250mM, 300mM, 350mM, 400mM, 450mM, 500mM, 600mM, 700mM, 800mM, 900mM most. Optionally, the concentration may be: (a) up to 1 μM, 2μM, 3μM, 4μM, 5μM, 10μM, 15 μM, 20 μM, 25 μM, 30 μM; (b) within a group selected range consisting of 35-335 μM, 40-300 μM, 50-300 μM, 50-250 μM, 50200 μM, 60-300 μM, 60-250 μM, 60-200 μM , 80-300 µM, 80-250 µM, 80-200 µM, 100 to 300 µM, 100 to 250 µM, or 100 to 200 µM; or (c) at least, or about, 345 µM, 350 µM, 360 µM, 370 µM, 380 µM, 390 µM, 400 µM, 450 µM, 0.5 mM, 1 mM, 2 mM or more.
[264] Optionally, the concentration may be within a range selected from the group consisting of about 1μM to about 1 mM, or about 30μM to about 0.5 mM, or about 60 μM to about 0 .4 mM.
[265]Atopic dermatitis (AD) affects 10-20% of children with 60% of cases occurring in the child's first year and 85% before the age of 5 (Krakowski, et al. Pediatrics, 122(4):812- 24 (2008)). Many cases persist into adulthood as evidenced by the 1-3% prevalence of AD among the adult population (Leung, et al., Lancet, 361(9352):151-60 (2003)). AD patients are colonized with S. aureus and this organism has been shown to exist both on dry skin and in areas of severe dermatitis (Ikezawa, et al.,Allergy Asthma Immunol Res.,2(4):235-46 ( 2010)). Disease severity has been directly correlated with the degree of S. aureus colonization, and therapy generally does not improve symptoms in the presence of high S. aureus counts (Akiyama, et al., J Dermatol Sci., 23(3): 155-6 (2000)). Confocal laser scanning microscope demonstrated the presence of biofilms in skin peeling and biopsy samples from AD patients (Akiyama et al. Br J Dermatol.,148(3):526-32 (2003)). The presence of S. aureus biofilms has been shown in samples of bullous impetigo and pemphigus foliaceus (Akiyama, et al., Br J Dermatol., 148(3):526-32 (2003)) while biofilms containing both S. aureus and Streptococcus pyogenes has been identified in non-bullous impetigo (Akiyama, et al., J Dermatol Sci.,32(3):193-9 (2003)). The difficulty in eradicating S. aureus colonization with conventional antibiotic therapy may be due to the presence of biofilms. Biofilm formation was also demonstrated in a mouse model inoculated with S. aureus isolated from a boil (Yamasaki et al., J Antimicrob Chemother.,48(4):573-7 (2001)).
[266] Biofilms have been implicated in miliaria by a clinical study in which only the extracellular polymer substance (EPS) producing S. epidermidis was able to induce lesions after inoculation and occlusion (Mowad, et al., J Am Acad Dermatol. , 33(5 Pt 1):729-33 (1995)). Biopsy samples revealed sweat glands blocked with EPS material, further supporting a pathogenic role for biofilms in this condition. Several factors, for example, the firm adhesion of dermatophytes to the nail plate, the presence of latent fungal elements, the ability of yeast to form biofilms and the difficulty of eradication, suggest the involvement of the biofilm in onychomycosis (Burkhart et al. J Am Acad Dermatol., 47(4):629-31 (2002 )).
[267] Chronic wounds present an ideal environment for microbial proliferation. In a clinical study of 66 wounds of various etiologies, 60% of chronic wounds were shown to contain biofilms, compared to 6% of acute wounds, indicating a role for biofilms in wound chronicity. Traditional cultures have identified Staphylococcus, Pseudomonas, and Enterococcus as the predominant organisms (James, et al., Wound Repair Regen.,16(1):37-44 (2008).
[268] In a preferred embodiment, the compounds may be incorporated into wound irrigation solutions. In another preferred embodiment, the compounds can be incorporated into cosmetic formulations.
[269] Compositions of compounds according to the third aspect of the present invention are also useful in oral health for both the prophylaxis and treatment of infections. For example, the compounds can be used to treat or prevent infections of the dental pulp by Streptococcus anginosus, or to prevent biofilms from binding to tooth surfaces. The compounds can be applied directly to tooth surfaces or applied to the dental pulp during a procedure. The compounds may also be incorporated into dental products, such as toothpaste, mouthwashes, dental floss, toothpicks, and chews (including food products), a mouthguard, a dental instrument, dentures, dental retainers, dental appliances. , including plastic appliances (such as Invisalign), toothbrush bristles, dentures and orthodontic devices, chewable non-food items or foods, as well as applied as coatings directly to dental tissues. The compositions can be used for the dental care of humans and animals, including pets, such as dogs and cats, as well as cattle and horses. For example, the compounds can be incorporated into chewable foods or toys, such as dog bones and biscuits.
[270] Indeed, in an embodiment of particular interest to the present invention, there is provided a human or animal (especially dog) chewable composition comprising one or more compounds as defined in Section III.A. Exemplary dog and other animal chewers that can be modified to include one or more compounds as defined in Section III.A include those described in US Patent 6,086,940, the contents of which are incorporated herein by reference. Other exemplary chewers include the Oravet® dental hygiene chewer produced by Merial (see http://merial.com/en/press-releases/merial-introduces-oravet-dental-hygiene-chews-for-dogs/, the contents of which are incorporated herein by reference) and KaNoodles dental chews (see http://kanoodlesusa.com/, the contents of which are incorporated herein by reference). Dental chews in accordance with the present invention can be used in dogs and other animals to inhibit the production of plaque forming biofilms and/or to reduce or treat or prophylactically treat halitosis. Chewing on such chewers can also help to ward off existing rubbing plaques and/or stones. Optionally, the chewers can usefully be used regularly, such as daily, and optionally, daily, after one or more meals.
[271] The compounds may, in accordance with the third aspect of the present invention, be added to drinking water or other drinking fluids.
[272] Other modes of administration according to the third aspect of the present invention may include: (i) Parenteral administration, which may include administration to a patient intravenously, intradermally, intraarterially, intraperitoneally, intranasally, intracranially, intraarticular, intraprostatic, intrapleural, intratracheal, intravitreal, intratumoral, intramuscular, subcutaneous, subconjunctival, intravesicular, intrapericardial, intraumbilical, by injection and by infusion, for example, as further described in Section III.C.1 of this application, below. Parenteral administration may include the use of formulations, such as described herein, that are formulated for controlled release, including immediate release, delayed release, extended release, pulsatile release, and combinations thereof, as further described in section III.C.1. (a) of this order, below. (ii) The compounds can be incorporated into injectable/implantable solid or semi-solid implants, such as polymeric implants, for example as described below in Section III.C.1 (b) of this application, below. (iii) Enteral administration, including administration in the form of suitable oral dosage forms such as tablets, capsules, solutions, suspensions, syrups and lozenges, for example, as described further in Section III.C.2 of this application , below. Optionally, enteral administration may include administration of controlled release enteral formulations, including oral dosage forms, such as capsules, tablets, solutions and suspensions, which are formulated for controlled release, including extended and/or delayed release, as described above. in more detail below in Section III.C.2(a) of this order. (iv) The administration of one or more disinfectant formulations or cleaning formulations, such as those described in Section III.C.4 of this application, below. 3. Hospital and other environments
[273] The methods and uses of the third aspect of the present invention may be practiced in the hospital and also in other medical and non-medical settings in order to address, inhibit, treat, ameliorate and/or disrupt biofilms. Other examples of microbial infection and biofilm colonization and formations that may be addressed by the third aspect of the invention are discussed further below and also further define medical uses and methods in accordance with the third aspect of the present invention for the treatment and/or prophylaxis of individuals. (including humans and animals) in need thereof.
[274] For example, S. epidermidis contributes to biofilms that grow on plastic devices placed inside the body (Otto, Nature Reviews Microbiology, 7(8):555-567 (2009)). This occurs most commonly in intravenous catheters and medical prostheses (Hedin, Scandinavian Journal of Infectious Diseases Supplementum, 90:1-59 (1993)). Infection can also occur in dialysis patients or anyone with an implanted plastic device that may have become contaminated. Another disease it causes is endocarditis. This occurs more often in patients with defective heart valves. In some other cases, sepsis can occur in hospitalized patients.
[275] As a further example, Methicillin-resistant S. aureus (MRSA), is one of a number of much-feared strains of S. aureus that have become resistant to most e-lactam antibiotics. Strains of MRSA are most often found associated with institutions such as hospitals, but are becoming increasingly prevalent in community-acquired infections. A recent study by the Translational Genomics Research Institute showed that nearly half (47%) of meat and poultry in US warehouses were contaminated with S. aureus, with more than half (52%) of these bacteria resistant to antibiotics (ScienceDaily, April 15, 2011).
[276] In another example, Enterococcus faecalis causes many of the antibiotic-resistant infections in hospitals as a result of its inherent resistance to certain antibiotics and its ability to survive and proliferate in the intestinal tract. Escherichia coli is one of the most frequent causes of many common bacterial infections, including cholecystitis, bacteremia, cholangitis, urinary tract infections and other clinical infections such as neonatal meningitis and pneumonia. For example, the compositions can be used to treat (e.g., as adjunctive therapy) conditions caused by community and/or hospital-acquired urinary tract infections (ICU) caused by strains of Escherichia coli (drug resistant or not) in immunocompromised patients.
[277] According to another example, aggressive colonization of stainless steel surfaces by P. aeruginosa, for example, is not only of enormous industrial importance but also of medical relevance; P. aeruginosa infections are prevalent in burn units where large stainless steel tubs, known as hydrotherapy units, are often used to treat patients with severe burns.
[278] Antibiotics are very ineffective in removing biofilms, although the third and second aspects of the present invention can be combined to potentiate the effect of antibiotics.
[279] The most common treatment for these infections is to remove or replace the infected implant, although in all cases prevention is ideal. The drug of choice is usually Vancomycin, to which rifampin or aminoglycoside may be added. Hand washing has been shown to reduce the spread of infection. Accordingly, the compositions according to the third aspect of the present invention may include hand washing and/or hand spraying compositions, and may accordingly be used accordingly in the treatment of hands and other organs.
[280] Preliminary research also indicates that S. epidermidis is found universally within acne-affected pores vulgaris, where Propionibacterium acnes is normally the sole resident (Bek-Thomson, et al., J. Clin. Microbiol., 46(10)) :3355-3360 (2008). The. Use as a disinfecting agent
[281] One or more compounds for use in the third aspect of the present invention may, in another embodiment, be used as disinfecting agents (or pesticides) (the United States Environmental Protection Agency, "EPA", defines biofilms as pestilents), for example in high-risk environments such as in hospital equipment or healthcare facilities. As such, the one or more compounds may be formulated in the form of a disinfecting formulation or cleaning formulation, such as those described in Section III.C.4 of this application, below.
[282] In accordance with another embodiment of the third aspect of the present invention, there is provided a method or use which comprises using the disinfecting agent in high risk environments, such as in hospital equipment or healthcare facilities, cosmetics , consumer or industrial applications, to prevent biofilm build-up or reduce biofilm from a surface of interest. In these embodiments, the compounds may, for example, be sprayed onto the surface in the form of a foam, solution or gel, or applied to the (clean) surface by means of a carrier, e.g. fabric, material or other porous article. containing one or more compounds. Another embodiment of the third aspect of the present invention is a disinfecting agent, as described herein, and there is also provided a product or article treated with a disinfecting agent, as described herein.
[283]The World Health Organization (WHO) estimates that, at any given time, more than 1.4 million people worldwide are affected by hospital-acquired infections. Cleaning, disinfection and sterilization save lives and improve patient outcomes. Between 5% and 10% of patients admitted to modern hospitals in developed countries acquire one or more healthcare-associated infections. The Centers for Disease Control and Prevention (CDC) estimates that approximately 1.7 million healthcare-associated infections occur annually in hospitals in the United States, and are associated with approximately 100,000 deaths annually. Healthcare-associated infections are also a major problem in long-term care facilities, including nursing homes and rehabilitation facilities. Transmission of healthcare-associated pathogens most often occurs through the hands of healthcare workers, who inadvertently contaminate their hands during various patient care activities. Less often, contaminated surfaces in healthcare facilities can contribute to the spread of healthcare-associated pathogens.
[284] The different levels of disinfection used in a health care facility can be defined by the Spaulding Classification (Sehulster, et al., Guidelines for the Control of Environmental Infections in Health Care Facilities. Recommendations from the CDC and the Health Infection Control Practices Advisory Committee (HICPAC). Chicago IL; American Health Engineering Society/American Hospital Association; 2004.). Spaulding levels, non-critical, semi-critical, and critical, are based on the potential for infectious disease to spread through equipment, instruments, and furniture, as well as the level of sterility typically required for the body part that comes in contact with it. The disinfection levels that correlate with the Spaulding classification are low, intermediate, high, and sterile. The US Centers for Disease Control (CDC) has further outlined disinfection levels for environmental surfaces in their "Guidelines for Environmental Infection Control in Health Care Facilities".
[285] Critical items carry a high risk of infection if contaminated with any microorganism. Thus, the third aspect of the present invention also provides objects treated for sterilization, as described herein, such objects enter sterile tissue or vascular system and must be sterile because any microbial contamination can transmit disease. This category includes surgical instruments, cardiac and urinary catheters, implants and ultrasound probes used in sterile body cavities. Semi-critical items contain mucous membranes or non-intact skin. This category includes respiratory therapy and anesthesia equipment, some endoscopes, laryngoscope blades, esophageal manometry probes, cystoscopes, anorectal manometry catheters, and diaphragm adjustment rings. These medical devices must be free of any microorganisms; however, small amounts of bacterial spores are permissible. Specific examples of critical or semi-critical instruments include invasive endoscopes such as laparoscopes and rigid instruments without an operating channel. Arthroscopes and laparoscopes that are inserted into sterile body cavities, as well as ancillary instrumentation, must be sterile. Other examples include gastroscopes, duodenoscopes, sigmoidoscopes, proctoscopes, colonoscopes, bronchoscopes, and laryngoscopes.
[286] The compounds can also be used in accordance with the third aspect of the present invention as food processing aids. For example, solutions of one or more compounds as defined in section III.A below may be sprayed onto animal carcasses or products (including meat part products) derived therefrom (i.e. poultry, fish and meat). or others, for example as described in connection with the first aspect of the present invention) to prevent or inhibit bacterial colonization or inactivate biofilm formation. The compounds may, for example, be applied by immersing chicken (or other animal) carcasses or a product derived therefrom in a container of a solution of the compounds, or by spraying an animal carcass with a solution of the compounds.
[287] In a preferred embodiment, aqueous solutions of FeQ, FeTyr, FeDOPA and/or Fe-Phe can be used as food processing aids. After treatment, the compounds can, if desired, be removed by washing.
[288] Another embodiment of the third aspect of the present invention provides an animal carcass (such as a chicken or other poultry, fish or other meat) and/or products (including meat part products) derived therefrom which have been treated, for example by spraying or dipping, in accordance with the third aspect of the present invention, and optionally wherein the one or more compounds are subsequently completely or partially removed by washing. B. Use as a coating
[289] In other embodiments of the third aspect of the present invention, one or more compounds having the structure of Formula A or B, or other compounds of the invention of the present invention as described further in Section III.A of this application, can be incorporated into coatings used to coat medical devices and other articles.
[290] Accordingly, the third aspect of the present invention also provides a method of coating a device or other article, comprising applying a coating comprising, consisting essentially of, or consisting of, one or more compounds having the structure of Formula A or B, or other compounds of the present invention, as described further in section III.A of this application.
[291] The third aspect of the present invention also provides coated devices or articles, having a coating comprising, consisting essentially of, or consisting of, one or more compounds having the structure of Formula A or B, or other compounds of the invention of the present invention. as described further in Section III.A of this application.
[292] Suitable coating methods are known in the art. Methods for coating medical devices are disclosed, for example, in Publications US20030054090 and 20120276280 and US Patents 5,879,697, 7,247,338 and 8,028,646. The compounds can be applied to medical devices and other articles in a variety of ways, including, but not limited to, ionic binding to a surface coating, passive adsorption or dispersion within a polymeric base material that forms the surface of the device or coated onto the surface. device surfaces (e.g. by dip coating, spray coating, ultrasonic spray coating, melt processing, filming, solvent coating, etc.).
[293] In a preferred embodiment, the one or more compounds are combined with polymers and coated onto medical devices or other articles. Suitable polymers include, but are not limited to, poly(lactides); poly(glycolides); poly(lactide-co-glycolide); polylactic acid); poly(glycolic acid); poly(lactic acid-co-glycolic acids); polycaprolactones; poly(orthoesters); polyanhydrides; poly (phosphazenes); polyhydroxyalkanoates [including poly-3-hydroxybutyrate, poly-3-hydroxybutyrate-co-3-hydroxyvalerate (PHBV), poly-4-hydroxybutyrate, poly-3-hydroxybutyrate-co-4-hydroxybutyrate]; synthetically or biologically prepared polyesters (including polyesters with one or more of the following monomeric units: glycol, lactic; trimethylene carbonate, p-dioxanone, or ε-caprolactone); poly(lactides-co-caprolactones); polyesters; polycarbonates; tyrosine polycarbonates; polyamides (including synthetic and natural polyamides, polypeptides and poly(amino acids)); polyesteramides; poly(dioxanones); poly(alkylene alkylate); polyethers (such as polyethylene glycol, PEG and polyethylene oxide, PEO); polyvinylpyrrolidones or PVP; polyurethanes; polyethesters; polyacetals; polycyanoacrylates; poly(oxyethylene)/poly(oxypropylene) copolymers; polyacetals, polyketals; polyphosphates; (containing phosphorus); polyphospho esters; polyalkylene oxalates; polyalkylene succinates; poly(maleic acids); chitin; chitosan; modified chitosan; collagen; silk; biocompatible polysaccharides; biocompatible copolymers (including block copolymers or random copolymers); hydrophilic or water-soluble polymers, such as polyethylene glycol, (PEG) or polyvinylpyrrolidone (PVP), with blocks of other biocompatible or biodegradable polymers, e.g. poly(lactide), poly(lactide-co-glycolide or polycaprolcatone or combinations thereof , polymers and copolymers of ethylene and propylene, including ultra high molecular weight polyethylene, ultra high molecular weight polypropylene, nylon, polyesters such as poly(ethylene terephthalate), poly(tetrafluoroethylene), polyurethanes, poly(ethylene urethanes), poly(methylmethacrylate), polyether ether ketone, polyolefins, dacron, latex, silicones, polymeric cements and poly(ethylene oxide).
[294] In another preferred embodiment of the third aspect of the present invention, one or more compounds may first be conjugated to other agents that have an affinity for, or may react with, a surface, and thus immobilized on a surface. For example, compounds can be linked to a bond that can be photoactivated to bind to a surface, or activated through another mechanism.
[295] Examples of devices and articles that can be coated using the compositions include tubing and other surface medical devices such as urinary catheters, stents, mucosal extraction catheters, suction catheters, umbilical cannula, contact lenses, intraoral devices. uterine, intravaginal and intraintestinal devices, endotracheal tubes, bronchoscopes, dental prostheses and orthodontic devices, surgical instruments, dental instruments, tubing, dental water lines, dental drainage tubes, tissue, paper, indicator strips (e.g., paper indicator strips or plastic indicator strips), adhesives (eg hydrogel adhesives, hot adhesives or solvent-based adhesives), bandages, tissue dressings or healing devices and occlusive patches and any other surface devices used in the medical field. Devices can include electrodes, external prostheses, fixation tapes, compression bandages and monitors of various types. Medical devices also include any device that can be placed at the site of insertion or implantation, such as the skin near the site of insertion or implantation, and that includes at least one surface that is susceptible to colonization by microorganisms embedded in biofilm. In a specific embodiment, a composition is integrated into an adhesive, such as a tape, thereby providing an adhesive which can prevent the growth or proliferation of biofilm-embedded microorganisms on at least one surface of the adhesive. Medical devices include equipment surfaces in operating rooms, emergency rooms, hospital rooms, clinics and bathrooms. In a particularly preferred embodiment, the following devices may be coated with the compounds: catheters, including central venous catheters, urinary catheters, dialysis catheters, and indwelling catheters (e.g., catheters for hemodialysis and for administering chemotherapeutic agents), cardiac implants, including mechanical heart valves, stents, ventricular assist devices, pacemakers, cardiac rhythm management devices (CRM), cardiac resynchronization therapy (CRT) devices and implantable cardioverter defibrillators (ICDs), synthetic vascular grafts, arteriovascular shunts, cerebrospinal fluid shunts, cochlear devices, joint prostheses, orthopedic implants, internal fixation devices, bone cements, percutaneous sutures, surgical mesh and surgical corrections including hernia repair meshes and adhesives, breast reconstruction meshes and adhesives, meshes and stickers for breast and face lifts, slings and meshes and patches for pelvic floor reconstruction, tracheal and ventilation tubes, dressings, biological implants (including allografts, xenografts and autografts), penile implants, intrauterine devices, endotracheal tubes, and lenses of contact.
[296] Other articles that can be coated in accordance with the third aspect of the present invention include articles for use on farm animals, such as animals and articles mentioned in the context of the first aspect of the present invention.
[297] However, other articles that may be coated in accordance with the third aspect of the present invention include articles for use in the process of slaughtering and/or processing animal carcasses or parts, such as animals and articles mentioned in the context of the first aspect of the present invention.
[298] Still other articles that can be coated in accordance with the third aspect of the present invention include articles for the preparation and/or containment of foodstuffs, including foodstuffs comprising raw or cooked meat, eggs, dairy products or other food products. The food products may be human and/or animal food products.
[299] Still other articles that may be coated in accordance with the third aspect of the present invention include articles for preparing and/or containing beverages.
[300] Accordingly, in another embodiment of the third aspect of the present invention there is provided a method of disinfecting a surface, or protecting a surface from infection, in need thereof, the method comprising contacting the surface with an amount of one or more compounds having the structure having Formula A or B, or other compounds of the invention of the present invention, as further described in Section III.A of the present application, wherein the one or more compounds are coated onto the surface to be disinfected.
[301] In some embodiments, the one or more compounds may be applied to the surface in the form of a spray, aerosol, or foam.
[302]The coated surface may, for example, be formed on the surface of an instrument selected from the group consisting of surgical instruments, cardiac and urinary catheters, implants, and ultrasound probes used in sterile body cavities.
[303]The surface may, for example, be formed on the surface of a device selected from the group consisting of urinary catheter, stents, mucosal extraction catheter, suction catheter, umbilical cannula, contact lenses, intrauterine devices, intravaginal devices, and intra-intestinal tubes, endotracheal tubes, bronchoscopes, prostheses and orthodontic devices, surgical instruments, dental instruments, tubing, dental water lines, dental drainage tubes, tissue, paper, indicator strips (e.g. paper indicator strips or plastic indicator strips) , adhesives (e.g. hydrogel adhesives, hot melt adhesives or solvent-based adhesives), bandages, tissue dressings or healing devices and occlusive plasters, central venous catheters, urinary catheters, dialysis catheters and indwelling catheters, heart implants, mechanical heart valves, stents, ventricular assist devices, pacemakers, d cardiac rhythm management devices (CRM), cardiac resynchronization therapy devices (CRTs) and implantable cardioverter defibrillators (ICDs), orthopedic implants, internal fixation devices, bone cements, percutaneous sutures, surgical mesh and surgical patches including repair mesh hernia mesh, breast reconstruction mesh, breast and face lift mesh, slings and mesh for pelvic floor reconstruction, tracheal and ventilatory tubes, wound dressings, biological implants, penile implants, intrauterine devices, endotracheal tubes, and contact.
[304]The coated surface may, for example, be formed on the surface of an article selected from the group consisting of industrial piping, liquid distribution lines, oil and gas pipelines, and cosmetic containers.
[305]The coated surface may, for example, be formed on the surface of, or be incorporated into, or on top of, a household item, such as an item selected from the group consisting of household disinfectants; laundry detergent; cleaning products; equipment involved in the leaching or mining process; wound care; toothpaste; mouthwashes; floss; toothpicks; chewable products (including foodstuffs); a mouthguard; a dental instrument; dentures; dental retainers; dental appliances, including plastic appliances (such as Invisalign); toothbrush bristles; dental prostheses and orthodontic appliances; non-food chewable items, food or toys, such as dog bones and cookies; a vacuum system; HVAC systems ((heating, ventilation and air conditioning)); vacuum cleaner bags; paint coating; wall coverings; window frames; doors; door frames; cooling towers; humidifiers; vacuum cleaners; filters such as a vacuum filter, a humidifier filter, a hot tub filter, or a pool filter; toys; plastic bottles; water jugs; faucet and water nozzle; washing machines; dishwashers; water dishes for animals; bathroom tiles and accessories; sinks; showers; Sanitary; sanitary covers; toilet seats; sealants and mortar; towels; TUPPERWARE®; plates; cups; utensils such as forks, spoons, knives and spatulas; bowls; food storage containers; beverage storage containers; cutting boards; drying trays; trash bags; sinks; fish ponds; pools; liners for swimming pools; pool skimmer; pond liners; bird baths; garden hose; sprinkler water lines; planters; and hot tubs.
[306]The coated surface may, for example, be formed on the surface of an article, device or apparatus used in the rearing and/or transport of animals, such as a chicken, for example, a meat type chicken, such as broilers , or a laying hen, such as a pullet or a hen, or a broiler, other poultry, such as a turkey, geese, quail or ducks, or livestock, such as cattle, sheep, goats or swine, alpaca, banteng, bison, camel, cat, deer, dog, donkey, llama, mule, rabbit, reindeer, water buffalo, yak, although one skilled in the art may appreciate that other animal foods, including zoo animals, animals in captivity , game animals, fish (including freshwater and saltwater fish, farmed fish and ornamental fish), other marine and aquatic animals, including crustaceans such as oysters, mussels, clams, shrimp, lobsters, crayfish , crabs, cuttlefish, octopus and squid, domestic animals animals such as cats and dogs, rodents (such as mice, rats, guinea pigs, hamsters) and horses, as well as any other domestic, wild and farmed animal, including mammals, marine animals, amphibians, birds, reptiles, insects and other invertebrates. In some embodiments, the device or apparatus used in raising and/or transporting animals may be selected from an article, device or apparatus which is for the distribution and/or containment of animal feed and/or animal drinking water.
[307]The coated surface may, for example, be formed on the surface of an article, device or apparatus used in the rearing and/or transport of animals, such as a chicken, for example a meat type chicken, such as broilers , or a laying hen, such as a pullet or a hen, or a broiler, other poultry, such as a turkey, geese, quail or ducks, or livestock, such as cattle, sheep, goats or swine, alpaca, banteng, bison, camel, cat, deer, dog, donkey, llama, mule, rabbit, reindeer, water buffalo, yak, although one skilled in the art may appreciate that other animal foods, including zoo animals, animals in captivity , game animals, fish (including freshwater and saltwater fish, farmed fish and ornamental fish), other marine and aquatic animals, including crustaceans such as oysters, mussels, clams, shrimp, lobsters, crayfish , crabs, cuttlefish, octopus and squid, domestic animals animals such as cats and dogs, rodents (such as mice, rats, guinea pigs, hamsters) and horses, as well as any other domestic, wild and farmed animal, including mammals, marine animals, amphibians, birds, reptiles, insects and other invertebrates. In some embodiments, the article, device or apparatus used in the rearing, housing and/or transport of animals may include one or more than one article, device or apparatus used in the production, rearing, collection, storage, processing and/or packaging of a animal product. For example, an animal product may be a by-product of the animal (eg, milk, eggs, or wool) or a product downstream thereof. Alternatively, an animal product may be the body or body part of the animal and the harvesting process optionally includes the step of slaughtering the animal and further optionally preparing an animal carcass or part thereof as a product, such as a meat product. .
[308] The third aspect of the present invention therefore also provides a device, article, product, item, formulation, composition or coating per se, having a coating comprising one or more compounds having the structure or having the structure of Formula A or B, or other inventive compounds of the present invention as described further in section III.A of this application, and for their use in the methods defined above.
[309] In one embodiment, the device, article, product, item, formulation, composition or coating comprises one or more compounds in the coating in an amount effective to prevent biofilm formation. In one embodiment, the device, article, product, item, formulation, composition or coating comprises one or more compounds in the coating in an amount effective to treat or reduce biofilm formation.
[310] The third aspect of the present invention also provides the direct products per se of the above-defined methods and uses of the third aspect of the present invention, and downstream product produced therefrom.
[311] The third aspect of the present invention also provides a compound conjugated to a structure that can anchor to a surface, wherein the compound has the structure of having the structure of Formula A or B, or other compounds of the invention of the present invention such as described in section III.A of this order. It may be preferred that the compound is selected from the group consisting of a complex of an amino acid or an α-hydroxy acid with Fe III, such as a complex of quinic acid with Fe III, a complex of L-tyrosine with Fe III , a complex of L-DOPA with Fe III, and a complex of L-phenylalanine with Fe III. Optionally, the compound may be selected from the group consisting of compounds represented by Formula I, Formula II, Formula III, Formula IV, Formula V, Formula VI, Formula VII, Formula VIII, Formula IX, Formula X, Formula XI, Formula XII , formula XIII, in Formula XIV, a compound that binds to major outer membrane proteins (MOMPs) or FlaA Campylobacter, a synthetic human histo-blood group antigen, a human histo-blood group antigen mimetic, or a synthetic sugar .
[312] Also provided by the third aspect of the present invention is a composition comprising one or more conjugated compounds as defined above, and an article coated with one or more of said conjugated compounds, or said composition.
[313] In one embodiment, the structure of the conjugated compound comprises hydroxyapatite or a derivative thereof, and the conjugate is capable of anchoring, or is anchored to, dental tissue.
[314] For example, in an additional embodiment, conjugated forms of the compounds, such as those shown in Figures 16A and B, in which the compounds are conjugated to hydroxyapatite, can be applied to tooth tissues, such as tooth enamel, dentin and pulp, caries and infection. In another embodiment, the compounds can be applied using photoreactive chemistry, for example, using conjugated forms of the compounds, such as those shown in Figures 15A and B. 4. Industrial, cosmetic and consumer applications
[315] The compositions can be used in accordance with another embodiment of the third aspect of the present invention to disinfect industrial surfaces, preventing and/or removing biofilm buildup on those surfaces. In this embodiment, biofilm formation can be prevented or inhibited, or a pre-formed biofilm can be removed by a method comprising applying a composition of the present invention comprising one or more compounds having the structure of Formula A or B, or other compounds of the present invention, as described further in section III.A of this application, on a surface in need thereof, for example in the form of a spray, foam, gel, powders; dishwashing or laundry detergents (liquid or solid), surface wax, window cleaners, etc.
[316] Therefore, the third aspect of the present invention also provides an object or article that has been treated in accordance with the above method.
[317] Biofilms are continuously produced and often accumulate on numerous industrial and biological surfaces. In an industrial environment, the presence of these biofilms causes a decrease in the efficiency of industrial machinery, requires increased maintenance and presents potential health risks. For example, the surfaces of water cooling towers become increasingly coated with microbiologically produced biofilm sludge, which reduces water flow and reduces heat exchange capacity. Water cooling tower biofilms can also harbor pathogenic microorganisms such as Legionella pneumophila. Food preparation lines are routinely plagued by biofilm buildup both on the machine and on the food product where biofilms often include potential pathogens. Biofilm formation comes with associated problems, such as accelerated equipment deterioration through corrosion of cellular byproducts. There may also be a reduction in the efficiency of heat transfer and a decrease in sensing devices as the film stops transmission.
[318] Pseudomonas aeruginosa readily attaches to stainless steel or plastic surfaces (eg poly(vinyl chloride), polystyrene) causing major problems in both the medical and food industries, forming biofilm. Biofilms easily form on PVC and glass surfaces under static conditions, especially in the food industry. The. industrial applications
[319] The compositions and coatings according to the third aspect of the present invention can be used to clean or maintain pipes and hoses in industries such as food and beverage industries, paper mills, sewage treatment, drainage, cooling towers and oil and gas industries coming into contact with a surface with biofilm growth with the composition. Industrial applications include its use in dairy lines, either as a rinse or wash for such lines, or incorporated into lines, for example, as a coating; liquid distribution lines in the manufacture or distribution of food and beverages, for example, use as a coating on feed lines for high distribution of sugar or syrup in the manufacture of soft drinks; pulp and paper mills (for biofouling); in the manufacture and containment of cosmetics from production line equipment to the final consumable, embedded within the cosmetic or coated onto the bottle containing the cosmetic; in water treatment facilities; in the leaching process used in mining; to prevent corrosion caused or accelerated by organisms, in oil and gas pipelines, including fracturing pipes, in oilfield acidification, in antifouling coatings (e.g. on submarines and boats) and in cooling towers. B. Consumer and light commercial applications
[320] Consumer and light commercial uses of the compounds and coatings according to the third aspect of the present invention include their incorporation into general household disinfectants; laundry detergent; cleaning products; equipment involved in the leaching or mining process; wound care; a vacuum system; HVAC systems (heating, ventilation and air conditioning); vacuum cleaner bags; paint coating; wall coverings; window frames; doors; door frames; cooling towers; boat hulls, humidifiers; vacuum cleaners; filters and membranes, such as a vacuum filter, a humidifier filter, hot tub filter, osmosis membranes, or a pool filter; toys; plastic bottles; water jugs; toothpaste, mouthwash, faucet and water nozzle; incorporation into plastics for a variety of household items, including inside and outside washing machines and dishwashers; water dishes for animals; bathroom tiles and accessories; sinks; showers; showers; Sanitary; sanitary covers; toilet seats; sealants and mortar; towels; TUPPERWARE®; plates; cups; utensils such as forks, spoons, knives and spatulas; bowls; food storage containers; beverage storage containers; cutting boards; drying trays; trash bags; bathtubs, including whirlpools and whirlpools; sinks; ponds and fish tanks swimming pools; liners for swimming pools; skimmer for swimming pools; pond liners; bird baths garden hose; sprinkler water lines; planters; and hot tubs. ç. cosmetic applications
[321] Another embodiment of the third aspect of the present invention provides cosmetics and cosmetic applications, as well as cosmetic containers and cosmetic applicators that incorporate and/or are coated with one or more compounds having the structure of Formula A or B, or others. inventive compounds of the present invention as described further in section III.A of this application.
[322] Cosmetics (also known as makeup or make-up) include care substances used to improve the appearance or odor of the human body. They are usually mixtures of chemical compounds, some derived from natural sources (including natural oils) and many are synthetic. A cosmetic can be a substance that is suitable to be applied to the human body to cleanse, beautify, promote attractiveness or alter appearance without affecting the body's structure or functions. While soap is not traditionally considered a cosmetic, for the purposes of the present description, the discussion of cosmetics can also apply to soaps.
[323] Exemplary cosmetics include skin creams, lotions, powders, perfumes, lipsticks, nails, eye and face makeup, towels, permanent waves, colored contact lenses, hair dyes, hair sprays and gels. , deodorants, hand sanitizer, products, bath oils, bubble baths, bath salts, butters and many other types of products. A subset of cosmetics is called "makeup", which primarily refers to colored products intended to alter the wearer's appearance. Cosmetics that are intended to be used on the face and eye area are usually applied with a brush or fingertips.
[324] Cosmetics can comprise a variety of organic and inorganic compounds. Typical organic compounds may include modified natural oils and fats as well as a variety of petrochemically derived agents. Inorganic compounds can include processed minerals such as iron oxides, talc and zinc oxide. Zinc and iron oxides can be classified as pigments, ie dyes, and may not have solubility in solvents.
[325]Application of the third aspect of the present invention to cosmetics, cosmetic applications, cosmetic containers and/or cosmetic applicators can provide methods for reducing, preventing, minimizing or disrupting biofilms on cosmetics, containers and/or applicators. Furthermore, as the application of the cosmetic to the user's body achieves the release of one or more compounds having the structure of Formula A or B, or other compounds of the invention of the present invention as described further in section III.A of this application, then cosmetics can be used to treat individuals in accordance with any of the embodiments of the second to third aspects of the present invention, particularly in the context of treating, reducing, preventing or stopping bacterial infections, colonization or biofilms on the skin, hair, nails and/or teeth of the user. 5. Additional medical applications
[326] In another embodiment of the third aspect of the present invention, compounds having the structure of Formula A or B, or other compounds of the invention of the present invention as described further in Section III.A of this application, and compositions comprising one or more of said compound can be used to treat any medical condition associated with biofilm formation as a result of microorganisms including, but not limited to gram-negative and gram-positive bacteria, including Pseudomonas, H. pylori, E. feacalis , Campylobacter, E. coli, EPEC, UPEC and Staphylococcus.
[327] In addition to the conditions discussed above, rarer but more serious manifestations of MRSA such as necrotizing fasciitis and pyomyositis (most commonly found in the tropics), necrotizing pneumonia, infective endocarditis (affecting the heart valves), and bone infections may occur. and articulate. Additional conditions include severe or extensive disease (eg, involving multiple sites of infection) or rapid progression in the presence of associated cellulitis, signs and symptoms of systemic disease, associated comorbidities or immunosuppression, extremes of age, abscess in an area that is difficult to drain, face, hand, and genitalia), associated septic phlebitis and unresponsiveness to incision and drainage alone, purulent cellulitis, hospitalized patients with complicated SSTI (cSSTI, defined as patients with deeper soft tissue infections, surgical infection, and ulcers and burns infected), osteomyelitis, device-related osteoarticular infections.
[328] In another embodiment, compounds having the structure of Formula A or B, or other compounds of the invention of the present invention, as described further in Section III.A of this application, and compositions comprising one or more of said compound, they can also be used to treat keratitis, colon cancer (where biofilms play a role) and peri-implantitis, a bacterial infection around an implant that results in inflammation of the gums and can lead to bone loss in the jaw.
[329] Certain strains of enterohemorrhagic E. coli (EHEC) found in the intestines of animals and humans can cause disease and can be life-threatening in a small group of patients who develop hemolytic hemorrhagic syndrome (HUS). EHEC is not treated with antibiotics because of the risks of developing HUS. The compounds may be useful in the treatment of EHEC infections, both in humans and animals, and particularly in livestock.
[330]Uropathogenic E. coli (UPEC) is the predominant etiologic agent causing UTIs. Accordingly, the compositions can also be used to inhibit or reduce the biofilm involved in lower urinary tract infections (UTIs). Human UTI's have traditionally been considered a self-limiting disease involving bacteria residing in the lumen of the bladder. Community-like intracellular bacterial structures were also identified in the urine sediments of patients with UTIs in a prospective study.
[331] In one embodiment, the biofilm that is inhibited or disrupted by the third aspect of the present invention may be a bacterial biofilm. The bacteria that form the biofilm can be gram-positive, or in an alternative embodiment they can be gram-negative, or the biofilm can be formed by a mixture of gram-positive and gram-negative bacteria.
[332] Optionally, the biofilm can be formed by bacteria selected from the group consisting of S. epidermidis, E. faecalis, E. coli, S. aureus, H. pylori, Campylobacter, Enteropathogenic Escherichia coli (EPEC), Uropathogenic Escherichia coli (UPEC), and Pseudomonas or combinations thereof. Optionally, in certain embodiments of the third aspect of the present invention, the biofilm is a biofilm that is formed by bacteria that are not bacteria that comprise, essentially consist of, or consist of a class of proteobacteria, such as any one or more of the spiruloid Wolinella spp. , Helicobacter spp., and more particularly Campylobacter spp.
[333] Optionally, the one or more compounds administered to a subject (such as a human or animal) according to the third aspect of the present invention may be a pharmaceutical or veterinary product and may further include one or more excipients, such as discussed in Section III. C of the present application, below.
[334] In an embodiment of the third aspect of the present invention, for treating biofilms in a subject (such as a human or animal), the one or more compounds is administered to a subject by one or more selected routes of delivery: parenteral as discussed below in section III.C.1 of this application, including a controlled release formulation as discussed below in section III.C.1(a) of this application, and injectable or implantable formulation as discussed below in section III.C.1 (b) of this order; enteral delivery, as discussed below in section III.C.2 of this application, including a controlled release enteral formulation, as discussed below in section III.C.2(a) of this application, with further reference to dosage forms of extended-release and delayed-release dosage forms as discussed therein; oral administration; topical administration, as discussed below in section III.C.3 of this application, including as an emulsion, lotion, cream, ointment, gel or foam, as discussed in parts (a), (b), (c), (d) ) (e) and (f) respectively below in section III.C.3 of this order; buccal distribution; sublabial distribution; sublingual distribution; in or on a dental product, such as a toothpaste, a mouthwash, a dental floss, a mouthguard; dermal distribution; or transdermal delivery.
[335] In some embodiment of the third aspect of the present invention, the biofilm may be associated with a bacterial infection selected from the group consisting of impetigo, boils, abscesses, folliculitis, cellulitis, necrotizing fasciitis, pyomyositis, surgical/traumatic wound infection, and infected ulcers and burns), osteomyelitis, device-related osteoarticular infections, impetigo, secondarily infected skin lesions, meningitis, brain abscess, subdural empyema, spinal epidural abscess, arterial damage, gastritis, urinary tract infections, biliary tract infections, pyelonephritis , cystitis, sinusitis, ear infections, otitis media, otitis externa, leprosy, tuberculosis, conjunctivitis, bloodstream infections, benign prostatic hyperplasia, chronic prostatitis, lung infections, including chronic lung infections of humans with cystic fibrosis, osteomyelitis, catheter infections, bloodstream infections, infections skin diseases, acne, rosacea, dental caries, periodontitis, gingivitis, nosocomial infections, arterial damage, endocarditis, periprosthetic joint infections, open or chronic wound infections, venous stasis ulcers, diabetic ulcers, arterial leg ulcers, pressure ulcers , endocarditis, pneumonia, infections of orthopedic prosthesis and orthopedic implants, peritonitis from peritoneal dialysis, cirrhosis and any other acute or chronic infection that involves or has a biofilm.
[336] Another embodiment of the third aspect of the present invention provides a method of treating a microbial infection in a subject in need thereof, the method comprising administering to the subject an effective amount of one or more compounds having the structure of Formula A or B, or other inventive compounds of the present invention as described further in section III.A of this application. Likewise, this embodiment also provides for the use of one or more of said compounds for the treatment of a microbial infection in a subject in need thereof.
[337] In certain embodiments, the microbial infection is caused by bacteria, such as gram positive bacteria or gram negative bacteria. For example, the infection may be caused by bacteria selected from the group consisting of S. epidermidis, E. faecalis, E. coli, S. aureus, H. pylori, Campylobacter, Enteropathogenic Escherichia coli (EPEC), uropathogenic Escherichia coli (UPEC). ), and Pseudomonas or combinations thereof and/or optionally, wherein the infection is not caused by bacteria that comprise, consist essentially of, or consist of a class of Proteobacteria, such as any one or more of Wolinella spp., Helicobacter spp., and more particularly Campylobacter spp.
[338] Optionally, in treating a microbial infection in a subject in need thereof in accordance with this embodiment of the third aspect of the present invention, the one or more compounds can be administered to a subject by parenteral delivery; enteric delivery; oral delivery; topical administration, such as in the form of an emulsion, lotion, cream, ointment, gel or foam; buccal distribution; sublabial distribution; sublingual distribution; in or on a dental product or dental device, such as a dental product, including but not limited to a toothpaste, a mouthwash, a dental floss, toothpicks, chewable products (including food products), a mouthguard , a dental instrument, dentures, dental retainers, dental appliances, including plastic appliances (such as Invisalign), toothbrush bristles, dentures and orthodontic devices, non-food chews, food or toys, such as dog and cookies; dermal distribution; or transdermal distribution.
[339] In certain embodiments, treatment of a bacterial infection in a subject in need thereof in accordance with this embodiment of the third aspect of the present invention may be to treat an infection selected from the group consisting of impetigo, boils, abscesses, folliculitis, cellulitis, necrotizing fasciitis, pyomyositis, surgical/traumatic wound infection, and infected ulcers and burns), osteomyelitis, device-related osteoarticular infections, impetigo, secondarily infected skin lesions, meningitis, brain abscess, subdural empyema, spinal epidural abscess, damage arterial infections, gastritis, urinary tract infections, biliary tract infections, pyelonephritis, cystitis, sinusitis, ear infections, otitis media, otitis externa, leprosy, tuberculosis, conjunctivitis, bloodstream infections, benign prostatic hyperplasia, chronic prostatitis, infections lung infections, including chronic lung infections of humans with cystic fibrosis a, osteomyelitis, catheter infections, bloodstream infections, skin infections, acne, rosacea, dental caries, periodontitis, gingivitis, nosocomial infections, arterial damage, endocarditis, periprosthetic joint infections, open or chronic wound infections, stasis ulcers venous ulcers, diabetic ulcers, arterial leg ulcers, pressure ulcers, endocarditis, pneumonia, orthopedic prosthetic and orthopedic implant infections, peritoneal dialysis peritonitis, cirrhosis and any other acute or chronic infection that involves or has a biofilm.
[340] In certain embodiments for treating a microbial infection in a subject in need thereof in accordance with this embodiment of the third aspect of the present invention, the infection may be caused by a strain of E. coli.
[341] Optionally, the treatment of a microbial infection in a subject in need thereof in accordance with this embodiment of the third aspect of the present invention may be for the treatment of a urinary tract infection.
[342] Optionally, treating a microbial infection in a subject in need thereof In accordance with this embodiment of the third aspect of the present invention, the subject may be one who is hospitalized and/or is immunocompromised.
[343] Optionally, treatment of a microbial infection in a subject in need thereof in accordance with this embodiment of the third aspect of the present invention may also include the additional administration of one or more antimicrobial agents, such as one or more antibiotics, to the subject. This may, for example, be conducted in accordance with any one or more of the embodiments of the second aspect of the present invention. III. COMPOUNDS AND COMPOSITIONS
[344]The present inventors have identified a class of a wide variety of activity, particularly against bacteria, and have developed numerous methods and uses involving the compounds, particularly in the formation of compositions. Compounds, which are further defined in Section III.A of this application, below, and compositions comprising one or more of said compounds, are presented herein as a fourth aspect of the present invention. Compounds and compositions comprising one or more of the compounds can be used to inhibit or reduce biofilm formation on a surface, treat or prevent an infection, and kill some antibiotic resistant organisms. In one embodiment, the invention is generally directed to compounds and compositions comprising one or more of the compounds, and methods and uses that employ one or more of the compounds and/or compositions, to inhibit, reduce, or prevent biofilm formation or accumulation in a surface or to remove, disperse, reduce or eradicate biofilm on a surface. In another embodiment, the invention also relates generally to compounds and compositions comprising one or more of the compounds, and methods and uses that employ one or more of the compounds and/or compositions, for the treatment of, growth inhibition. and inhibition of colonization by bacteria, both in biological and non-biological environments. In another embodiment, the invention also relates to compounds and compositions comprising one or more of the compounds, and methods and uses that employ one or more of the compounds and/or compositions, for disinfecting surfaces in both biological and non-biological environments, and products that have been coated or treated by one or more of the compounds and/or compositions of the present invention. In another embodiment, the invention also relates to compounds and compositions comprising one or more of the compounds, and methods and uses that employ one or more of the compounds and/or compositions, to potentiate the effects of one or more antibiotics, increasing the sensitivity of bacteria (including antibiotic-resistant bacteria) to one or more antibiotics, and also to reverse antibiotic resistance in bacteria. In yet another embodiment the invention also relates to compounds and compositions comprising one or more of the compounds, and to methods and uses which employ one or more of the compounds and/or compositions, to enhance the growth of animals and their efficiency of use. in foods, in particular by oral administration of food and beverage compositions. A. Compounds
[345] The following compounds as described in this section of the application are provided herein as a fourth aspect of the present invention.
[346] All other aspects of the present invention may utilize one or more types of compounds as defined in this section, including derivatives and salts as defined in subsections 1 and 2, respectively.
[347] Compositions comprising, consisting essentially of, or consisting of, one or more of these compounds are also provided as another embodiment of the fourth aspect of the present invention. These compositions may be used in all other various aspects of the present invention, and methods and uses of the present invention which employ said compositions, and may comprise, consist essentially of, or consist of, one or more types of compounds as defined herein. section, including derivatives and salts, as defined in subsections 1 and 2, respectively.
[348] Without limitation, compounds of particular interest for use in accordance with the present invention include Fe III complexes comprising ligands attached to the iron center selected from amino acids or α-hydroxy acids, including but not limited to ferric quinate (also herein interchangeably referred to as FeQ and Fe-QA), ferric tyrosine (also referred to herein as FeTyr), ferric DOPA (also referred to herein as FeDOPA), and ferric phenylalanine (also referred to herein as Fe-Phe). Furthermore, compounds that are structural and/or functional variants, derivatives and/or analogs of the foregoing compounds, as described later in this section, are of particular interest to the present invention.
[349] Ligands that can be used in these complexes include ligands based on amino acids, α-hydroxy acids, o-hydroxybenzoic acids or pyridine-2-carboxylic acids.
[350] Examples of amino acids may include, but are not limited to, alanine, arginine, asparagine, aspartic acid, cysteine, glutamine, glutamic acid, glycine, histidine, isoleucine, leucine, lysine, methionine, phenylalanine, proline, serine, threonine, tryptophan, tyrosine and valine, each preferably in the L isoform, although, as discussed above, in an alternative embodiment one or more (optionally all) may be in the D isoform. Mixtures of optical isomers of the same amino acid may or may not be used in some modalities.
[351]Exemplary α-hydroxy acids include, but are not limited to, quinic acid, lactic acid, glycolic acid, citric acid, and mandelic acid.
[352]Exemplary o-hydroxybenzoic acids include, but are not limited to, salicylic acid.
[353]Exemplary pyridine-2-carboxylic acids include, but are not limited to α-picolinic acid.
[354] In certain embodiments, compounds for use in the present invention, and which can bind Campylobacter MOMPs or FlaA, are Fe III complexes each containing three bidentate ligands, as described herein.
[355] In further embodiments, compounds for use in the present invention, and which may optionally bind to MOMPs or Campylobacter FlaA, are Fe III complexes defined according to the following chemical formula A:
Formula A or a salt and/or hydrate thereof, wherein: X, X1 and X2 may independently be NH2, OH, CO2-, CO2H, OR3, NR3H, NR3R4, R3ONO2, R3NO2, SH, SR3, and X, X1 and X2 can all be the same or they can all be different, or alternatively, two can be the same and one can be different; Y, Y1 and Y2 can independently be O, NH, NH2, NR3, NR3R4, SH, OR3, OH, and Y, Y1 and Y2 can be the same or they can all be different, or alternatively, two can be the same and one may be different; Z, Z1 and Z2 can independently be: O, S, NH, NR3 , and Z, Z1 and Z2 can be the same or they can all be different, or alternatively, two can be the same and one can be different; R, R', R1, R1', R2, and R2' may independently be H, CH3, CH2SH, CH2CO2H, CH2CH2CO2H, CH2C6H5, CH2C3H3N2, CH(CH3)CH2CH3, (CH2)4NH2, CH2CH(CH3)2, CH2CH2SCH3 , CH2CONH2, (CH2)4NHCOC4H5NCH3, CH2CH2CH2, CH2CH2CONH2, (CH2)3NHC(NH)NH2, CH2OH, CH(OH)CH3, CH2SeH, CH(CH3)2, CH2C8H6N, CH2C6H4OH and R, R', R1, R1' , R2 , and R2' can all be the same or they can all be different, or alternatively, up to five can be the same and one or more can be different; or any relevant pair of R and R', R1 and R1', and R2 and R2' (i.e. when they are attached to the same carbon atom) are linked to form a substituted or unsubstituted cycloalkyl ring group; R3 and R4 may independently be alkyl, alkenyl, alkynyl, phenyl, aryl, hydroxy- or halo-substituted radicals, hydroxyl radicals, nitrogen-substituted radicals, oxygen-substituted radicals, or hydrogen. In some embodiments, R3 and R4 may all be the same or they may all be different, or alternatively, two may be the same and one may be different.
[356] In embodiments in which one or more pairs of R and R', R1 and R1', and R2 and R2' are linked to form a substituted or unsubstituted cycloalkyl ring group, the substituents on the cycloalkyl group may be selected from, but are not limited to, =O and particularly OH, NH2 , NR3 , NR3 R4 , SH, and OR3 ; wherein R3 and R4 are defined as above.
[357] Bonds between Fe and X, X1 and X2 and between Fe and Y, Y1 and Y2 are preferred to be ionic.
[358] In a particular embodiment X, X1 and X2 may independently be NH2, OH, CO2-, CO2H, OR3, NR3H or NR3R4 (preferably NH2 or OH); Y, Y1 and Y2 may independently be O, NH, NH2, OR3 or OH (preferably O); Z, Z1 and Z2 may independently be O or S (preferably O); R, R', R1, R1', R2, and R2' may independently be H, CH3, CH2SH, CH2CO2H, CH2CH2CO2H, CH2C6H5, CH2C3H3N2, CH(CH3)CH2CH3, (CH2)4NH2, CH2CH(CH3)2, CH2CH2SCH3 , CH2CONH2, (CH2)4NHCOC4H5NCH3, CH2CH2CH2, CH2CH2CONH2, (CH2)3NHC(NH)NH2, CH2OH, CH(OH)CH3, CH2SeH, CH(CH3)2, CH2C8H6N, CH2C6H4OH; or any relevant pair of R', R1 and R1', and R2 and R2' are linked to form a substituted or unsubstituted 4- to 6-membered cycloalkyl ring group, optionally wherein the substituents on the cycloalkyl group are selected from = O and, in particular, OH, NH2, NHR3, NR3R4, SH, and OR3); and R3 and R4 independently represent methyl, ethyl, propyl, butyl or benzyl.
[359] Particular compounds that may be mentioned include those wherein R', R1' and R2' represent H, and R, R1 and R2 represent a group as defined above other than H; or each pair of R and R', R1 and R1', and R2 and R2' (i.e. when they are attached to the same carbon atom) are linked to form an optionally substituted cyclohexyl ring group, one or more substituents selected from =O and particularly, OH, NH2, NHR3, NR3R4, SH, and OR3.
[360] In a further embodiment, R3 and R4 are independently C1-4 alkyl, C1-4 alkenyl, phenyl or benzyl (whose last four groups are optionally substituted by one or more halo or hydroxyl groups). For example, R3 and R4 may independently represent methyl, ethyl, propyl, butyl or benzyl.
[361] In a preferred embodiment, Y, Y1 and Y2 represent O, and Z, Z1 and Z2 represent O. Particular examples of such compounds include those wherein X, X1 and X2 independently represent NH2 or OH.
[362] Functional variants of compounds according to Formula A may also be used in the present invention, and include other compounds as described in this section of the application.
[363] For example, in a still or additional embodiment, compounds for use in the present invention and which can bind Campylobacter MOMPs or FlaA are Fe III complexes defined according to the following chemical formula B: Formula B
or a salt and/or hydrate thereof, wherein: X3, X4 and X5 may independently be -C(R8)=, or -N=; R8 may independently be NH2, OH, CO2-, CO2H, OR9, NR9H, NR9R10, R9ONO2, R9NO2, SH, SR9, and each R8 may all be the same or may all be different, or alternatively, two may be the same and one may be different; Y3, Y4 and Y5 can independently be O, NH, NH2, NR9, NR9R10, SH, OR9, OH and Y3, Y4 and Y5 can all be the same or they can all be different, or alternatively, two can be the same and one may be different; Z3, Z4 and Z5 can independently be: O, S, NH, NR9, and Z3, Z4 and Z5 can all be the same or they can all be different, or alternatively, two can be the same and one can be different; m1, m2 and m3 may independently be 0, 1, 2, 3 or 4; and m1, m2 and m3 can all be the same or they can all be different or alternatively two can be the same and one can be different; R5, R6, and R7 are each independently selected from OH, NH2, NHR9, NR9R10, SH, and OR9; and R5, R6 and R7 may all be the same or they may all be different; R9 and R10 may independently be alkyl, alkenyl, alkynyl, phenyl, aryl, hydroxy- or halo-substituted radicals, hydroxyl radicals, nitrogen-substituted radicals, oxygen-substituted radicals, or hydrogen. In some embodiments, R9 and R10 may all be the same or they may all be different.
[364] It is preferred that the bonds between Fe and X3, X4 and X5 and between Fe and Y3, Y4 and Y5 are ionic. In a preferred embodiment, X3 , X4 and X5 may independently be -C(OH)=, or -N=; Y3 , Y4 and Y5 may independently O, NH, NH2 , OR9 or OH (preferably O); Z, Z1 and Z2 may independently be O or S (preferably O); R5 , R6 and R7 are each selected from OH, NH2 , NHR9 , and OR9 (preferably R5, R6 and R7 are all OH); M1, M2 and M3 can independently be selected from 0, 1 and 2; and R9 and R10 independently represent methyl, ethyl, propyl, butyl or benzyl.
[365] In a further embodiment, R9 and R10 are independently C1-4 alkyl, C1-4 phenyl or benzyl (whose last four groups are optionally substituted by one or more halo or hydroxyl groups). For example, R3 and R4 may independently represent methyl, ethyl, propyl, butyl or benzyl.
[366] In a preferred embodiment, Y3, Y4 and Y5 represent O, Z3, Z4 and Z5 represent O, R5, R6 and R7 represent OH, and m1, m2 and m3 are selected from 0, 1 and 2. Particular examples of such compounds include those wherein X3 , X4 and X5 independently represent -C(OH)= or -N=.
[367] In a preferred embodiment, the ligands attached to the iron core are amino acids or α-hydroxy acids. Therefore, it is more preferred that Y, Y1, Y2, Z, Z1 and Z2 represent O, X, X1 and X2 represent NH2 or OH, and R', R1' and R2' represent H. When one or more of the ligands is a amino acid (e.g. for compounds of formula A where X, X1 and X2 represent NH2), then it is preferred that the amino acid is an L-amino acid (or glycine), although in an alternative embodiment one or more (optionally all) of the ligands may be a D-amino acid. Examples of amino acids may include, but are not limited to, alanine, arginine, asparagine, aspartic acid, cysteine, glutamine, glutamic acid, glycine, histidine, isoleucine, leucine, lysine, methionine, phenylalanine, proline, serine, threonine, tryptophan, tyrosine and valine, each preferably in the L isoform, although, as discussed above, in an alternative embodiment one or more (optionally all) may be in the D isoform. Mixtures of optical isomers of the same amino acid may or may not be used in some modalities.
[368] Exemplary compounds of Fe complexes according to Formula A include Formulas VII-IX as shown below:
[369] g) a complex of L-DOPA with Fe III (3,4-dihydrophenylalanine) (Fe-DOPA)
Formula VII h) a complex of L-tyrosine with Fe III (Fe-Tyr, also called Fe-Y)
Formula VIII i) a complex of quinic acid with Fe III (Fe-QA, also called FeQ)
Formula IX
[370] Exemplary compounds of Fe complexes according to Formula B include Formulas X-XIV as shown below: j) a complex of 2,3,5-trihydroxybenzoic acid with Fe III
k) a complex of 2,4,5-trihydroxybenzoic acid with Fe
Formula XI l) a complex of 3-dehydroquinic acid with Fe III
m) a complex of 4,6-dihydroxypyridine-2-carboxylic acid with Fe III
n) a complex of salicylic acid with Fe III Formula XIV

[371] Optionally, in one embodiment, an Fe complex as described above (e.g., according to Formula A or Formula B) for use in any of the first, second, or third aspects of the present invention may not be a complex of quinic acid with Fe III (such as a complex having the structure of Formula IX). That is, in an optional embodiment, Formula A excludes a complex of quinic acid with Fe III (such as a complex having the structure of Formula IX).
[372] In a further embodiment, a compound according to Formula A, or Formula B, for use in the present invention may be a compound that inhibits binding of C. jejuni to a human histo-blood group antigen. This can, for example, be measured when the bacteria are grown in a medium containing the compound, the medium containing the compound is washed and the binding of the bacteria to the histo-blood group antigen is determined by an ELISA assay (as in accordance with the method described in Example 4) and compared to a control where the bacteria do not grow in the presence of the compound. Preferably, the compound inhibits C binding. jejuni to a histo-blood group antigen at a level that is at least about 1%, 2%, 3%, 4%, more preferably at or at least about 5%, even more preferably at , or at least about 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 95%, 96%, 97%, 98%, 99%, 100 % or more than the level of inhibition of C. jejuni binding to a histo-blood group antigen by an L-tyrosine complex with Fe III or a complex of quinic acid with Fe III at the same molar concentration.
[373] In another embodiment, a compound according to Formula A, or Formula B, for use in the present invention may be a compound that inhibits biofilm formation by bacteria, as measured in a plastic bead assay ( such as according to a method as described in Example 1), wherein the bacteria is grown in a medium containing the compound to form a growth suspension of the bacteria at 0.0001 OD/ml, the growth suspension is allowed to grow with plastic-coated UV beads (Lascells), and the beads are assayed after 24 hours for the presence of biofilm formation on the beads (counting bacteria after beads release) and compared to a control group in which the bacteria do not grow in the presence of the compound. Preferably, the compound inhibits the binding of bacteria to the plastic-coated beads to an inhibition level that is at least about 1%, 2%, 3%, 4%, more preferably by, or at least least about 5%, even more preferably at, or at least about 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 95%, 96 %, 97%, 98%, 99%, 100% or more of the level of inhibition of bacteria binding to plastic-coated UV beads by either an L-tyrosine complex with Fe III or a quinic acid complex with Fe III with the same molar concentration. In a particularly preferred embodiment, the bacteria may be Enterococcus faecalis, Staphylococcus epidermidis, Staphylococcus aureus, Campylobacter jejuni, Pseudomonas aeruginosa, Escherichia coli, Uropathogenic and Escherichia coli. enteropathogenic
[374] In another embodiment, a compound according to Formula A, or Formula B, for use in the present invention may be a compound that inhibits Helicobacter pylori binding to human gastric tissue (e.g., as determined by a method as described in Example 5) at a level of inhibition that is at least about 1%, 2%, 3%, 4%, more preferably at or at least about 5%, even more preferably at, or at least about 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 95%, 96%, 97%, 98%, 99%, 100% or greater than the level of inhibition of bacterial binding to human gastric tissue by an L-tyrosine complex with Fe III or a quinic acid complex with Fe III at the same molar concentration as measured by counting the average number of bacteria bound to the tissue.
[375] In another embodiment, a compound according to Formula A, or Formula B, for use in the present invention can be a compound that inhibits the biofilm formation of a bacterium, but does not inhibit the planktonic growth of the bacteria (e.g. example, as determined using a method as described in Example 7), wherein the bacteria can be one or more of the following: Enterococcus faecalis, Staphylococcus epidermidis, Staphylococcus aureus, Campylobacter jejuni, Pseudomonas aeruginosa, Uropathogenic Escherichia coli, and Escherichia coli. Enteropathogenic. Preferably, the compounds inhibit biofilm formation (e.g., as measured by the coverage rate in Example 7) at a level that is at or at least about 1%, 2%, 3%, 4% , more preferably at or at least about 5%, even more preferably at at least about 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 95%, 96%, 97%, 98%, 99%, 100% or more of the level of inhibition of biofilms by an L-tyrosine complex with Fe III or a quinic acid complex with Fe III with the same molar concentration.
[376] In another embodiment, a compound according to Formula A, or Formula B, for use in the present invention can be a compound for treating cystic fibrosis. In one embodiment, one or more compounds of Formula A can be administered using a mist spray. In one embodiment, one or more compounds of Formula A can be administered in liposomes for the treatment of patients with cystic fibrosis.
[377] In another embodiment, a compound according to formula A, or formula B, for use in the present invention can be a compound that prevents the attachment of bacteria to a surface (e.g., when determined in accordance with a method as described in Example 13), and preventing the bacteria from attaching to the surface is at a level that is at least about 1%, 2%, 3%, 4%, more preferably by, or at least about 5%, even more preferably at, or at least about 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 95% , 96%, 97%, 98%, 99%, 100% or more of the level of bacterial binding by an L-tyrosine complex with Fe III or a complex of quinic acid with Fe III at the same molar concentration as measured by optical density. In a particularly preferred embodiment, the bacteria may be Enterococcus faecalis, Staphylococcus epidermidis, Staphylococcus aureus, Campylobacter jejuni, Pseudomonas aeruginosa, Escherichia coli, Uropathogenic and Escherichia coli. Enteropathogenic.
[378] In another embodiment, a compound according to Formula A, or Formula B, for use in the present invention may be a compound that is capable of rendering an antibiotic resistant strain of bacteria sensitive to the antibiotic to which it is otherwise resistant mode (e.g., when determined by a method comprising immersing a patch in a solution of the compound and an antibiotic, such as kanamycin, e.g. at a concentration of 50 μg/mL as described in Example 9, placed on a plate with an antibiotic-resistant strain (such as an Enteropathogenic Escherichia coli or Campylobacter jejuni strain)), and causes the bacteria to fail to grow or to reduce the growth rate of the antibiotic-resistant strain in the presence of the antibiotic by a level that is one level which is at least about 1%, 2%, 3%, 4%, or at least about 5%, even more preferably at least about 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 95%, 96%, 97%, 98%, 99%, 100% or more than the level of growth rate reduction caused by a complex of L-tyrosine with Fe III or a complex of quinic acid with Fe III at the same molar concentration.
[379] In another embodiment, a compound according to Formula A, or Formula B, for use in the present invention can be a compound that causes a decrease in growth rate to a level that is, or at least, about 1%, 2%, 3%, 4%, more preferably at, or at least about 5%, even more preferably at, or at least about 10%, 20%, 30%, 40 %, 50%, 60%, 70%, 80%, 90%, 95%, 96%, 97%, 98%, 99%, 100% or more of the decrease in growth rate as measured by optical density of a resistant bacteria to antibiotics when grown in the presence of the compound and the antibiotic, for example, as determined by a method as described in Examples 11 and/or 12. The combinations of antibiotics and antibiotic-resistant bacteria can, for example, be one or more of following: (i) kanamycin and a kanamycin resistant bacterium, (ii) gentamicin and a gentamicin resistant Enteropathogenic Escherichia coli, and (iii) kanamycin and a clinical isolate Pseudomonas (PAO Clinical) as described in Example 14.
[380] According to one embodiment, instead of directly administering the one or more compounds, he or they may be formed in vivo, by administering a suitable iron-containing substance and one or more suitable ligands capable of forming the compounds in vivo. with the iron compound (see: Campbell and Hasinoff, Ferrous sulfate reduces levodopa bioavailability: Chelation as a possible mechanism, Clin. Pharmacol Ther. 45:220-5, 1989). For example, ferrous sulfate and tyrosine (as a ligand) can be administered to form Fe-Tyr in vivo, ferrous sulfate and L-DOPA (as a ligand) can be administered to form Fe-DOPA in vivo, ferrous sulfate and L-phenylalanine ( as ligand) can be administered to form Fe-Phe in vivo or ferrous sulfate and quinic acid (as ligand) can be administered from Fe-QA in vivo. In this example, Fe2+ is oxidized to Fe3+ in vivo, and can complex with tyrosine, L-DOPA, or phenylalanine, respectively. The compounds can also be formed in vivo from any substance that can be metabolized in vivo to the compounds. For example, phenylalanine can be administered with ferrous sulfate as it will be metabolized to tyrosine in vivo and can then complex with ferric iron (formed from the oxidation of ferrous sulfate). Alternatively, ferric chloride can also be administered with, for example, tyrosine, quinic acid, L-DOPA and/or phenylalanine.
[381] Optionally, one or more compounds for use in any of the first, second or third aspects of the present invention (which may or may not be compounds according to Formula A or Formula B as discussed above) are ligands for the proteins Outer membrane principals (MOMPs) or Campylobacter FlaA, and/or may be able to down-regulate the expression of FlaA and/or FlaB proteins in a bacterium such as Campylobacter, such as to the extent of causing reduced bacterial motility such as when determined by a method as described in Example 21 of the present application. Binding of compounds to MOMPs or FlaA inhibits the MOMPs or FlaA from binding, binding or associating with other proteins, biofilm components, surfaces or other bacteria.
[382] The compound may be a mimetic or synthetic human histo-blood group antigen or a synthetic sugar. A synthetic human histoblood group antigen may be a sugar, e.g. a saccharide having the same structure as a natural human histoblood group antigen, such as e.g. HI antigen, H-II antigen, Lewis antigen, Leb , Lex or Ley. A preferred compound is ferric quinate (Fe-QA).
[383] Compounds provided herein that bind to MOMPs or FlaA Campylobacter include compounds having the structures described in this section, according to Formulas A or B, or other compounds, as described below. These compounds have been shown to inhibit both gram negative bacteria such as Pseudomonas aeruginosa, Campylobacter jejuni, Helicobacter pylori, Escherichia coli, Enteropathogenic Escherichia coli (EPEC), Uropathogenic Escherichia coli (UPEC) and gram positive bacteria such as Staphylococcus epidermidis, Staphylococcus aureus, and Enterococcus faecalis, believed to be predictive of efficacy with other species There is low homology between Campylobacter MOMP and other bacteria. The compounds are believed to interact with several bacterial porin-like surface proteins that have not yet been identified in other bacteria.
[384] In additional embodiments, compounds for use in the present invention may or may not optionally include one or more compounds selected from: N-[3-quinylamino-2-(quinylaminomethyl)-propyl]-quinamide
b) N-{2-[Bis-(2-quinylaminoethyl)-amino]-ethyl}-quinamide
c) phosphoric acid tris-(2-ethyl-quinylamino) ester
d) N-(3,5-Bis-quinylamino-cyclohexyl)-quinamide
e) N-(4,5-Bis-quinylamino-2-hydroxy-6-hydroxymethyl-tetrahydropyran-3-yl)-quinamide
Formula V f) N-(4,5-bis-quinylamino-2-hydroxy-6-quinylaminomethyl-tetrahydropyran-3-yl)-quinamide Formula VI

[385] The foregoing compounds for use in any aspect of the present invention may also be in the form of hydrates or hydrate salts. For example, the compositions can be Fe-Tyr.xH2O, FeQ.xH2O, FeDOPA.xH2O or Fe-Phe.xH2O. The compounds may also be hydrates containing salts, for example hydrates with bases such as lithium hydroxide, sodium hydroxide or potassium hydroxide present.
[386]In the case of compounds that are Fe III complexes comprising ligands bound to the iron core as described above, in an option not all ligands will be the same in the compositions comprising the Fe III complex compounds. For example, in the case where the compound is FeTyr, then this may be formed by creating a complex of Fe III and a commercial source of tyrosine (Tyr), which may include low levels (typically less than 10%, such as less or more than 5% or about 2.5%) of one or more additional amino acids, such as cysteine (Cys) and/or phenylalanine (Phe), and so in an optional embodiment, when the compound is FeTyr, then some of the compounds in the composition may include one or more alternative amino acids (e.g., Cys and/or Phe) as ligands. The proportion of ligands in the FeTyr composition that are not Tyr can be less than 10%, 9%, 8%, 7%, 6%, 5%, 4%, 3%, 2% or 1% and can be substantially 0% . The same applies mutatis mutandis to other ligands used in the preparation of Fe III complexes for use in the present invention.
[387] Therefore, for example, in a composition comprising an Fe III complex as described above, it may be that less than 100% of the Fe III ligands are identical, although preferably at least 50 %, 60%, 70%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% of the ligands in the composition are identical . In this context, in one embodiment, the term "identical" discriminates between enantiomeric forms of ligand, i.e., different enantiomers are not identical; whereas, in another embodiment, the term "identical" may apply to different enantiomeric forms of ligand, i.e., optionally different enantiomeric forms of the same ligand are considered identical. 1. Derivatives
[388] Derivatives of compounds for use according to any aspect of the present invention, such as compounds defined above, including Formula I-IX, Formula X to XIV, Formula A (or hydrates thereof) and Formula B or hydrates of them) can also be used. The term "derivative" does not mean that the derivative is synthesized from the parent compound, either as a starting material or as an intermediate, although that may be the case. The term "derivative" can include salts (e.g. pharmaceutically acceptable salts), prodrugs or metabolites of the parent compound. Derivatives include compounds in which the free amino groups on the parent compound have been derivatized to form amine hydrochlorides, p-toluene sulfoamides, benzoxycarboamides, t-butyloxycarboamides, thiourethane-type derivatives, trifluoroacetylamides, chloroacetylamides or formamides. Derivatives include compounds having one or more amino substituents or hydrogen groups substituted with substituted or unsubstituted alkyl, aminoalkyl, aryl or heteroaryl groups having 1 to 30 carbon atoms. 2. Salts
[389] Compounds for use according to any aspect of the present invention, such as compounds defined above, including Formula I-IX, Formula X to XIV, Formula A (or the hydrates thereof) and Formula B or hydrates thereof) may be in the form of a salt, for example a pharmaceutically acceptable salt. Examples of pharmaceutically acceptable salts include, but are not limited to, mineral or organic acid salts of basic residues such as amines; alkali or organic salts of acidic residues such as carboxylic acids. Pharmaceutically acceptable salts include conventional toxic salts or quaternary ammonium salts of the parent compound formed, for example, from non-toxic organic or inorganic acids. Such conventional non-toxic salts include those derived from inorganic acids, such as hydrochloric, hydrobromic, sulfuric, sulfamic, phosphoric and nitric acids; and salts prepared from organic acids, such as acetic, propionic, succinic, glycolic, stearic, lactic, malic, tartaric, citric, ascorbic, pamoic, maleic, hydroxymaleic, phenylacetic, glutamic, benzoic, salicylic, sulfanilic acid, 2- acetoxybenzoic, fumaric, tolunesulfonic, naphthalenesulfonic, disulfonic, oxalic and isethionic ethane, and bases such as lithium hydroxide, sodium hydroxide, potassium hydroxide and ammonium hydroxide.
[390] Pharmaceutically acceptable salts of the compounds can be synthesized from the parent compound containing a basic or acidic moiety by conventional chemical methods. Generally, such salts can be prepared by reacting the free acid or base forms of these compounds with a stoichiometric amount of the appropriate base or acid in water or an organic solvent or a mixture of the two; generally, non-aqueous media such as ether, ethyl acetate, ethanol, isopropanol or acetonitrile are preferred. Lists of suitable salts are found in Remington's Pharmaceutical Sciences, 20th ed., Lippincott Williams & Wilkins, Baltimore, MD, 2000, p. 704; and "Handbook of Pharmaceutical Salts: Properties, Selection, and Use," P. Heinrich Stahl and Camille G. Wermuth Eds., Wiley-VCH, Weinheim, 2002. B. Antimicrobial agents
[391]Antimicrobial agents that can be used therapeutically and/or non-therapeutically with the compounds of the present invention according to any one of the first, second or third aspects of the present invention, for example for the treatment or prophylaxis of microbial infection in accordance with with the third aspect of the present invention and/or according to the second aspect of the present invention, either separately, simultaneously or sequentially, include, but are not limited to: (i) Aminoglycosides, including amikacin, gentamicin, kanamycin, neomycin, netilmycin , tobramycin, paromomycin, streptomycin, spectinomycin; (ii) Ansakines, including geldanamycin, herbimycin, rifaximin, (iii) Carbacephem, including loracarbef, (iv) Carbapenem, including ertapenem, doripenem, imipenem/cilastatin, meropenem, (v) Cephalosporins, including cefadroxil, cefazolin, cephalothin or cephalothin, Ceflandlor cefamandol cefoxitin cefrozil cefuroxime cefixima cefdinir cefotaxima cefodoxime ceftazidima ceftibutene cephetizoxima ceftriaxon cefepime ceftaroline fosamil ceftobiprol (vi) glycopeptides including teicoplanin, vancomycin, televancina , dalbavancin, oritavancin, (vii) Lincosamides, including clindamycin, lincomycin, (viii) lipopeptides including daptomycin, (ix) macrolides including azithromycin, clarithromycin, dirithromycin, erythromycin, roxithromycin, troleandomycin, telithromycin, spiramimine, (x) Monobactams, including aztreonam , (xi) Nitrofurans, including furazolidone, nitrofurantoin, (xii) Oxazolidinones, including linezolid, posizolid, radezolid, torezolid,(xiii) Penicillins, including amoxicillin, ampicillin, azlocillin, carbenicillin, cloxacillin, dicloxacillin, flucloxacillin, mezlocillin, methicillin, nafcillin, oxacillin, penicillin G, penicillin V, piperacillin, temocillin, ticarcillin, amoxicillin/clavulanate, ampicillin/sulbactam, piperacillin/tazobactam, ticarcillin/clavulanate (xiv) Polypeptides including bacitracin, colistin, polymyxin B, (xv) Quinolones/Fluoroquinolone, including ciprofloxacin, enoxacin, gatifloxacin, gemifloxacin, levofloxacin, lomefloxacin, moxifloxacin, nalidixic acid, norfloxacin ofloxacin, trovafloxacin, grepafloxacin, sparfloxacin, temafloxacin, (xvi) Sulfonamides, including mafenide, sulfacetamide, sulfadiazine, silver sulfadiazine, sulfadimetoxin, sulfametast, sulfamethoxazole, sulfanilamide, sulfasalazine, sulfisoxazole, trimethoprim-sulfamethoxazole (co-trimoxazole), (xvii) tetracyclines, including demeclocycline, doxycycline, minocycline, oxytetra cycline, tetracycline, (xviii) clofazimine, dapsone, capreomycin, cycloserine, ethambutol, ethionamide, isoniazid, pyrazinamide, rifampicin (rifampin), rifabutin, rifapentine, streptomycin, arsphenamine, chloramphenicol, fosfomycin, fusidic acid, metronidazole, mupirocin, platensimycin, quinupristin /dalfopristin, thiamphenicol, tigecycline, tinidazole and trimethoprim; and combinations thereof. The compounds can also be combined with triclosan and chlorhexidine. Other antimicrobial agents include: aztreonam; cefotetan and its disodium salt; loracarbef; cefoxitin and its sodium salt; cefazolin and I sodium salt; cefaclor; ceftibuten and its sodium salt; ceftizoxime; ceftizoxime sodium salt; cefoperazone and its sodium salt; cefuroxime and its sodium salt; cefuroxime axetil; cefprozil; ceftazidime; cefotaxime and its sodium salt; cefadroxil; ceftazidime and its sodium salt; cephalexin; nafate cefamandole; cefepime and its hydrochloride, sulfate and phosphate salt; cefdinir and its sodium salt; ceftriaxone and its sodium salt; cefixime and its sodium salt; cefpodoxime proxetil; meropenem and its sodium salt; imipenem and its sodium salt; cilastatin and its sodium salt; azithromycin; clarithromycin; dirithromycin; erythromycin and hydrochloride, sulfate or phosphate salts, ethylsuccinate and stearate, clindamycin; clindamycin hydrochloride, sulfate or phosphate salt; lincomycin and hydrochloride, sulfate or phosphate salt, tobramycin and its hydrochloride, sulfate or phosphate salts; streptomycin and its hydrochloride, sulfate or phosphate salts; neomycin and its hydrochloride, sulfate or phosphate salts; acetylsulfisoxazole; colistimethate and its sodium salt; quinupristine; dalphopristin; amoxicillin; ampicillin and its sodium salt; clavulanic acid and its sodium or potassium salt; penicillin G; benzathine penicillin G, or procaine salt; sodium or potassium salt of penicillin G; carbenicillin and its disodium or indanyl disodium salt; piperacillin and its sodium salt; ticarcillin and its disodium salt; sulbactam and its sodium salt; moxifloxacin; ciprofloxacin; ofloxacin; levofloxacins; norfloxacin; gatifloxacin; trovafloxacin mesylate; alatrofloxacin mesylate; trimethoprim; sulfamethoxazole; demeclocycline and its hydrochloride, sulfate or phosphate salts; doxycycline and its hydrochloride, sulfate or phosphate salts; oxytetracycline and its hydrochloride, sulfate or phosphate salts; chlorotetracycline and its hydrochloride, sulfate or phosphate salts; metronidazole; dapsone; atovaquone; rifabutin; linezolid; polymyxin B and its hydrochloride, sulfate or phosphate salts; sulfacetamide and its sodium salt; clarithromycin; and silver ions, salts and complexes.
[392] A preferred embodiment of any aspect of the present invention, such as according to the second aspect of the present invention, provides for the use of a complex of quinic acid with Fe III (Fe-QA, also called FeQ), such as defined by Formula IX, with any one or more of the foregoing antibiotics, formulated together in the same composition for administration or presented in separate compositions for use separately, simultaneously or sequentially.
[393] Another preferred embodiment of any aspect of the present invention, such as according to the second aspect of the present invention, provides for the use of a complex of L-tyrosine with Fe III (Fe-Tyr), as defined by Formula VIII, with any one or more of the foregoing antibiotics, formulated together in the same composition for administration or presented in separate compositions for use separately, simultaneously or sequentially.
[394] In another preferred embodiment, any aspect of the present invention, such as according to the second aspect of the present invention, provides for the use of a complex of L-DOPA with Fe III (3,4-dihydrophenylalanine) (Fe-DOPA), as defined by Formula VII, with any one or more of the foregoing antibiotics, formulated together in the same composition for administration or presented in separate compositions for use separately, simultaneously or sequentially.
[395] In another preferred embodiment of any aspect of the present invention, such as according to the second aspect of the present invention, provides for the use of a complex of L-phenylalanine with Fe III (Fe-Phe) with either or more of the above antibiotics, formulated together in the same composition for administration or presented in separate compositions for use separately, simultaneously or sequentially. C. Excipients and Carriers
[396] The compounds as defined in Section III.A above may be formulated for use in accordance with any of the first, second or third aspects of the present invention and may, for example, be formulated in a form that is suitable for administration enteral, parenteral, topical or pulmonary.
[397] The compounds as defined in section III.A above may be combined with one or more pharmaceutically acceptable carriers and/or excipients that are considered safe and effective and can be administered to an individual without causing undesirable biological side effects or unwanted interactions.
[398] The carrier may include all components present in the pharmaceutical formulation in addition to the active ingredient or ingredients. The compounds are included in the formulation in an amount effective to achieve the effect of the first, second or third aspects of the present invention, for example in an amount which is effective to inhibit biofilm formation or reduce biofilm accumulation. An effective amount of a compound provided to a subject can be an amount that is sufficient to provide the required degree of reduction in microbial colonization. This may depend on the type of compost and/or the size of the animal.
[399] In one embodiment, an effective amount of the compound can be an amount that is effective to deliver the compound to the site where action is required at a concentration ranging from 1μm to 1M, preferably greater than 10μM, 20μM , 30μm, 40 μm, 50 μm, 60 μm, 70 μm, 80 μm, 90 μm, 100 μm, 110 μm, 120μm, 130μm, 140 μm, 150 μm, 160 μm, 170 μm, 180 μm, 190 μm, 200 μM or more. A suitable concentration may be within the range of about 1 µm to about 1 mM, or about 30 µm to about 0.5 mM or about 60 µM to about 0.3 mM. These concentrations may particularly apply to the performance of the invention in the context of the second and/or third aspects of the present invention.
[400] In one embodiment an effective amount of the compound may be from 0.3 to 32 mg/day/kg of subject body weight, such as a chicken. In another embodiment, an effective concentration of the compound can range from 0.001 to 1 mm for use in coatings or devices, or solutions.
[401] The compounds may also be formulated for use as a disinfectant, for example, in a hospital setting or for industrial application. 1. Parenteral formulations
[402] Compounds as defined in section III.A above for use in accordance with any of the first, second or third aspects of the present invention and may be formulated for parenteral administration.
[403] Parenteral administration may include administration to a patient intravenously, intradermally, intraarterially, intraperitoneally, intralesionally, intracranially, intraarticularly, intraprostatically, intrapleurally, intratracheal, intravitreal, intratumoral, intramuscular, subcutaneous, subconjunctival, intravesicular , intrapericardial, intraumbilical, by injection and by infusion.
[404] Parenteral formulations can be prepared as aqueous compositions using techniques that are known in the art. Typically, such compositions may be prepared as injectable formulations, for example, solutions or suspensions; solid forms suitable for use in preparing solutions or suspensions after the addition of a reconstitution medium prior to injection; emulsions, such as water-in-oil (w/o) emulsions, oil-in-water (o/w) emulsions and microemulsions, liposomes or emulsions thereof.
[405] The carrier can be a solvent or dispersion medium containing, for example, water, ethanol, one or more polyols (for example, glycerol, propylene glycol, and liquid polyethylene glycol), oils, such as vegetable oils (for example, , peanut oil, corn oil, sesame oil, etc.) and combinations thereof. Proper flowability can be maintained, for example, by the use of a coating such as lecithin, by maintaining the required particle size in the case of dispersion, and by the use of surfactants. In many cases, it will be preferable to include isotonic agents, for example, sugars or sodium chloride.
[406]Solutions and dispersions of the active compounds such as the free acid or base or pharmacologically acceptable salts thereof may be prepared in water or in another solvent or dispersing medium suitably mixed with one or more pharmaceutically acceptable excipients including, but not limited to a, surfactants, dispersants, emulsifiers, pH modifying agents, viscosity modifying agents and combinations thereof.
[407] Suitable surfactants can be anionic, cationic, amphoteric or nonionic surfactants. Suitable anionic surfactants include, but are not limited to, those containing carboxylate, sulfonate and sulfate ions. Examples of anionic surfactants include sodium, potassium, long chain alkyl ammonium sulfonates and alkylaryl sulfonates such as sodium dodecylbenzene sulfonate; dialkyl sodium sulfosuccinates such as sodium dodecylbenzenesulfonate; dialkyl sodium sulfosuccinates such as sodium bis-(2-ethylthioxyl)sulfosuccinate; and alkyl sulfates, such as sodium lauryl sulfate. Cationic surfactants include, but are not limited to, quaternary ammonium compounds such as benzalkonium chloride, benzethonium chloride, cetrimonium bromide, stearyl dimethylbenzylammonium chloride, polyoxyethylene and coconut amine. Examples of non-ionic surfactants include ethylene glycol monostearate, propylene glycol myristate, glyceryl monostearate, glyceryl stearate, polyglyceryl-4-oleate, sorbitan acylate, sucrose acylate, PEG-150 laurate, PEG-400 monolaurate, polyoxyethylene monolaurate, polysorbates, polyoxyethylene octylphenylether, PEG-1000 cetyl ether, polyoxyethylene tridecyl ether, polypropylene glycol butyl ether, Poloxamer® (polyoxyethylene triblock copolymer, followed by a polyoxypropylene block, followed by a polyoxyethylene block) 401, stearoyl monoisopropanolamide and polyoxyethylene hydrogenated tallow amide. Examples of amphoteric surfactants include sodium N-dodecyl-β-alanine, sodium N-lauryl-β-iminodipropionate, myristoacetate, lauryl betaine and lauryl sulfobetaine.
[408]The formulation may contain a preservative to prevent the growth of microorganisms. Suitable preservatives include, but are not limited to, parabens, chlorobutanol, phenol, sorbic acid, and thimerosal. The formulation may also contain an antioxidant to prevent degradation of the active agent(s) or particles.
[409] The formulation is typically buffered to a pH of 3-8, for parenteral administration, by reconstitution. Suitable buffers include, but are not limited to, phosphate buffers, acetate buffers and citrate buffers. It should be noted that FeQ and some of the other compounds as defined in Section III.A of the application are acidic and are thus advantageously formulated with a buffer to achieve a suitable pH, particularly in the context of the preparation of the injectable formulation, including formulations for intravenous injection.
[410] Water-soluble polymers are often used in formulations for parenteral administration. Suitable water-soluble polymers include, but are not limited to, polyvinylpyrrolidone, dextran, carboxymethylcellulose and polyethylene glycol.
[411]Sterile injectable solutions can be prepared by incorporating the particles in the required amount in the appropriate solvent or dispersion medium, with one or more of the excipients listed above, as required, followed by filter sterilization. Generally, dispersions can be prepared by incorporating the various sterile active ingredients into a sterile vehicle that contains a basic dispersion medium and the other necessary ingredients from those listed above. In the case of sterile powders for the preparation of sterile injectable solutions, the preferred methods of preparation are vacuum-drying and freeze-drying techniques which produce a powder of the active ingredient plus any additional desired ingredient from a previously filtered sterile solution of the active ingredient. same. Powders can be prepared in such a way that the particles are porous in nature, which can increase the dissolution of the particles. Methods for preparing porous particles are well known in the art. (a) Controlled release formulations
[412]The parenteral formulations described herein comprising one or more compounds as defined in section III.A above for use in accordance with any of the first, second or third aspects of the present invention may be formulated for controlled release including immediate release, delayed release, extended release, pulsatile release, and combinations thereof. 1. Nano and microparticles
[413] For parenteral administration, the one or more compounds as defined in section III.A above for use in accordance with any of the first, second or third aspects of the present invention, and one or more optional additional active agents, may be incorporated into microparticles, nanoparticles or combinations thereof that provide controlled release of compounds and/or one or more additional active agents. In embodiments where the formulations contain two or more active components, such as drugs, then they may be formulated for the same type of controlled release (e.g., delayed, prolonged, immediate, or pulsatile) or they may be formulated independently for different types of drug. release (e.g. immediate and delayed, immediate and prolonged, delayed and prolonged, delayed and pulsatile, etc.).
[414] For example, the compounds and/or one or more additional active agents can be incorporated into polymeric microparticles, which provide controlled release of the active agent(s). The release of the active agent(s) is controlled by diffusion of the drug(s) out of the microparticles and/or degradation of the polymeric particles by hydrolysis and/or enzymatic degradation. Suitable polymers include ethylcellulose and other natural or synthetic cellulose derivatives.
[415] Polymers, which are slowly soluble and form a gel in an aqueous environment, such as hydroxypropyl methylcellulose or polyethylene oxide, may also be suitable as materials for drug-containing microparticles. Other polymers include, but are not limited to, polyanhydrides, poly(ester anhydrides), polyesters such as polylactide(PLA), polyglycolide (PGA), poly(lactide-co-glycolide) (PLGA), polydioxanone, poly-3 - hydroxybutyrate (PHB) and copolymers thereof, poly-4-hydroxybutyrate (P4HB) and copolymers thereof, polycaprolactone and copolymers thereof, but not limited to polymers of glycolic acid, lactic acid, 1,4-dioxanone, carbonate of trimethylene, 3-hydroxybutyric acid, 4-hydroxybutyrate, e-caprolactone, including polyglycolic acid, polylactic acid, polydioxanone, polycaprolactone, copolymers of lactic and glycolic acids, such as VICRYL® polymer, MAXON® and MONOCRYL® polymers, and including poly (lactide-co-caprolactones); poly (orthoesters); polyanhydrides; poly(phosphazenes); polyhydroxyalkanoates; synthetically or biologically prepared polyesters; polycarbonates; tyrosine polycarbonates; polyamides (including synthetic and natural polyamides, polypeptides and poly(amino acids)); polyetheramides; poly(alkylene alkylate); polyethers (such as polyethylene glycol, PEG, and polyethylene oxide, PEO); polyvinylpyrrolidones or PVP; polyurethanes; polyethesters; polyacetals; polycyanoacrylates; poly(oxyethylene)/poly(oxypropylene) copolymers; polyacetals, polyketals; polyphosphates; (containing phosphorus); polyphosphoesters; polyalkylene oxalates; polyalkylene succinates; poly(maleic acids); silk (including recombinant silks and silk derivatives and analogues); chitin; chitosan; modified chitosan; biocompatible polysaccharides; hydrophilic or water-soluble polymers, such as polyethylene glycol, (PEG) or polyvinylpyrrolidone (PVP), with blocks of other biocompatible or biodegradable polymers, e.g. poly(lactides), poly(lactides-co-glycolide, or polycaprolcatone and copolymers of the the same, including random copolymers and block copolymers thereof and combinations thereof.
[416]Alternatively, the active agent can be incorporated into microparticles prepared from materials that are either insoluble in aqueous solution or slowly soluble in aqueous solution, but are capable of degrading within the GI tract by means including enzymatic degradation, surfactant action of acids. bile and/or mechanical erosion. As used herein, the term "slowly soluble in water" refers to materials that do not dissolve in water within a 30 minute period. Preferred examples include fats, fatty substances, waxes, wax-like substances and mixtures thereof. Suitable fats and fatty substances include fatty alcohols (such as lauryl, myristyl, stearyl, cetyl, or cetostearyl alcohol), fatty acids and derivatives, including but not limited to esters of fatty acids, glycerides of fatty acids (mono-, di- and tri -glycerides) and hydrogenated fats. Specific examples include, but are not limited to, hydrogenated vegetable oil, hydrogenated cottonseed oil, hydrogenated castor oil, hydrogenated oils available under the tradename STEROTEX®, stearic acid, cocoa butter, and stearyl alcohol. Suitable waxes and wax-like materials include natural or synthetic waxes, hydrocarbons and normal waxes. Specific examples of waxes include beeswax, glucose wax, castor wax, carnauba wax, paraffins and candelilla wax. As used herein, a wax-like material is defined as any material, which is normally solid at room temperature and has a melting point of about 30 to 300°C.
[417] In some cases, it may be desirable to alter the rate of penetration of water into the microparticles. For this purpose, rate controlling (soaking) agents may be formulated together with the fats or waxes listed above. Examples of rate control materials include certain starch derivatives (e.g. waxy maltodextrin and drum dried corn starch), cellulose derivatives (e.g. hydroxypropyl methyl cellulose, hydroxypropyl cellulose, methyl cellulose and carboxymethyl cellulose), alginic acid, lactose and talc. Additionally, a pharmaceutically acceptable surfactant (e.g. lecithin) may be added to facilitate the degradation of such microparticles.
[418] Proteins, which are insoluble in water, such as zein, can also be used as materials for forming microparticles containing the active agent. Additionally, proteins, polysaccharides and combinations thereof, which are soluble in water, can be formulated with drug into microparticles and subsequently cross-linked to form an insoluble network. For example, cyclodextrins can be complexed with individual drug molecules and subsequently cross-linked. 2. Production method of nano and microparticles
[419]The encapsulation or incorporation of an active agent, such as one or more compounds as defined in section III.A above for use in accordance with any of the first, second or third aspects of the present invention, in carrier materials to produce microparticles containing drug can be achieved through known pharmaceutical formulation techniques. In the case of formulation in fats, waxes or wax-like materials, the support material is typically heated above its melting temperature and the active agent is added to form a mixture comprising particles of active agent suspended in the carrier material, active agent dissolved in the carrier material, or a mixture thereof. The microparticles may subsequently be formulated by a variety of methods, including, but not limited to, freezing, extrusion, spray-chilling or aqueous dispersion processes. In a preferred process, the wax is heated above its melting temperature, the active agent is added, and the molten wax-drug mixture is frozen under constant stirring as the mixture cools. Alternatively, the molten wax-drug mixture can be extruded and spheronized to form pellets or spheres. All these processes are known in the art.
[420] For some carrier materials, it may be desirable to use a solvent evaporation technique to produce microparticles containing the active agent. In this case, the active agent and the support material are co-dissolved in a mutual solvent and the microparticles can subsequently be produced by various techniques including, but not limited to, forming an emulsion in water or other suitable medium, spray drying or evaporating the solvent from the bulk solution and grinding the resulting material.
[421] In some embodiments, the particulate active agent is homogeneously dispersed in a water-insoluble or slowly water-soluble material. To minimize the size of the active agent particles within the composition, the active agent powder itself can be ground to fine particles prior to formulation. The jet milling process, known in the pharmaceutical art, can be used for this purpose. In some embodiments, the particulate active agent is homogeneously dispersed in a wax or wax-like substance by heating the wax or wax substance above its melting point and adding the active agent particles while stirring the mixture. In this case, a pharmaceutically acceptable surfactant can be added to the mixture to facilitate dispersion of the active agent particles.
[422]The particles may also be coated with one or more modified release coatings. Solid fatty acid esters, which are hydrolyzed by lipases, can be spray coated onto microparticles or active agent particles. Zein is an example of a naturally water-insoluble protein. It can be coated onto active agent-containing microparticles or active agent particles by spray coating or wet granulation techniques. In addition to naturally water-insoluble materials, some digestive enzyme substrates can be treated with crosslinking procedures, resulting in the formation of insoluble networks. Many methods of crosslinking proteins, initiated by chemical and physical means, have been reported. One of the most common methods for achieving crosslinking is the use of chemical crosslinking agents. Examples of chemical crosslinking agents include aldehydes (glutaraldehyde and formaldehyde), epoxy compounds, carbodiimides and genipin. In addition to these crosslinking agents, oxidized and native sugars were used to crosslink the gelatin. Crosslinking can also be carried out using enzymatic means; for example, transglutaminase has been approved as a GRAS substance for cross-linking seafood. Finally, crosslinking can be initiated by physical means such as heat treatment, UV irradiation and gamma irradiation.
[423] To produce a coating layer of cross-linked protein containing microparticles containing active agents or active agent particles, a water-soluble protein can be spray coated onto the microparticles and subsequently cross-linked by one of the methods described above. Alternatively, the active agent-containing microparticles can be microencapsulated within the protein by coacervation phase separation (e.g., by the addition of salts) and subsequently cross-linked. Some proteins suitable for this purpose include gelatin, albumin, casein, and gluten.
[424] Polysaccharides can also be cross-linked to form a water-insoluble network. For many polysaccharides, this can be achieved by reaction with calcium salts or multivalent cations, which cross-link the main polymer chains. Pectin, alginate, dextran, amylose and guar gum are subject to cross-linking in the presence of multivalent cations. Complexes can also be formed between oppositely charged polysaccharides; pectin and chitosan, for example, can be complexed via electrostatic interactions. (b) Injectable/implantable formulations
[425] The one or more compounds as defined in section III.A above for use in accordance with any of the first, second or third aspects of the present invention may be incorporated into injectable/implantable solid or semi-solid implants, such as polymeric implants. . In one embodiment, the compounds are incorporated into a polymer which is a liquid or paste at room temperature, but upon contact with an aqueous medium, such as physiological fluids, exhibits an increase in viscosity to form a semi-solid or solid material. Exemplary polymers include, but are not limited to, hydroxyalkanoic acid polyesters derived from the copolymerization of at least one hydroxyl unsaturated fatty acid copolymerized with hydroxyalkanoic acids. The polymer can be melted, mixed with the active substance and melted or injection molded in a device. Such melt fabrication requires polymers with a melting point that is below the temperature at which the substance to be released and the polymer degrade or become reactive. The device can also be prepared by solvent molding in which the polymer is dissolved in a solvent and the drug dissolved or dispersed in the polymer solution and the solvent is then evaporated. Solvent processes require the polymer to be soluble in organic solvents. Another process is compression molding of a mixed powder of the polymer and the drug or polymer particles loaded with the active agent.
[426]Alternatively, the compounds can be incorporated into a polymer matrix and molded, compressed or extruded in a device that is a solid at room temperature. For example, the compounds can be incorporated into a biodegradable polymer such as polyanhydrides, polyhydroalkanoic acids (PHAs), PLA, PGA, PLGA, polycaprolactone, polyesters, polyamides, polyorthoesters, polyphosphazenes, proteins and polysaccharides such as collagen, hyaluronic acid, albumin and gelatin, and combinations thereof and compressed into a solid device, such as discs, or extruded into a device, such as sticks. Other alternative polymers for use herein include polymers include, but are not limited to, polymers of glycolic acid, lactic acid, 1,4-dioxanone, trimethylene carbonate, 3-hydroxybutyric acid, 4-hydroxybutyrate, e-caprolactone, including polyglycolic, polylactic acid, polydioxanone, polycaprolactone, copolymers of glycolic and lactic acids, such as VICRYL® polymer, MAXON® and MONOCRYL® polymers, and including poly(lactide-cocaprolactones); poly (orthoesters); polyanhydrides; poly (phosphazenes); polyhydroxyalkanoates; synthetically or biologically prepared polyesters; polycarbonates; tyrosine polycarbonates; polyamides (including synthetic and natural polyamides, polypeptides and poly(amino acids)); polyesteramides; poly(alkylene alkylate); polyethers (such as polyethylene glycol, PEG and polyethylene oxide, PEO); polyvinylpyrrolidones or PVP; polyurethanes; polyethesters; polyacetals; polycyanoacrylates; poly(oxyethylene)/poly(oxypropylene) copolymers; polyacetals, polyketals; polyphosphates; (containing phosphorus); polyphosphoesters; polyalkylene oxalates; polyalkylene succinates; poly (maleic acids); silk (including recombinant silks and silk derivatives and analogues); chitin; chitosan; modified chitosan; biocompatible polysaccharides; hydrophilic or water-soluble polymers, such as polyethylene glycol, (PEG) or polyvinylpyrrolidone (PVP), with blocks of other biocompatible or biodegradable polymers, e.g. poly(lactides), poly(lactide-co-glycolide, or polycaprolkone and copolymers, including random copolymers and block copolymers thereof.
[427]The release of one or more compounds from the implant can be varied by polymer selection, polymer molecular weight, and/or polymer modification to enhance degradation, such as pore formation and/or incorporation of hydrolyzable bonds. Methods for modifying the properties of biodegradable polymers to vary the release profile of compounds from the implant are well known in the art. 2. Enteral formulations
[428] Compounds as defined in section III.A above for use in accordance with any of the first, second or third aspects of the present invention and may be formulated for parenteral administration.
[429] Suitable oral dosage forms include tablets, capsules, solutions, suspensions, syrups and lozenges. Tablets can be produced using compression or molding techniques well known in the art. Gelatin or non-gelatin capsules can be prepared as hard or soft capsule capsules, which can encapsulate liquid, solid and semi-solid fill materials, using techniques well known in the art.
[430]The formulations can be prepared using a pharmaceutically acceptable carrier. As generally used herein, "carrier" includes, but is not limited to, diluents, preservatives, binders, lubricants, disintegrants, swelling agents, fillers, stabilizers, and combinations thereof.
[431] The carrier also includes all components of the coating composition, which may include plasticizers, pigments, dyes, stabilizers and glidants.
[432] Examples of suitable coating materials include, but are not limited to, cellulose polymers such as cellulose acetate phthalate, hydroxypropylcellulose, hydroxypropylmethylcellulose, hydroxypropylmethylcellulose phthalate, and hydroxypropylmethylcellulose acetate succinate; polyvinyl acetate phthalate, acrylic acid polymers and copolymers, and methacrylic resins which are commercially available under the tradename EUDRAGIT® (Roth Pharma, Weiterstadt, Germany), zein, shellac and polysaccharides.
[433] In addition, the coating material may contain conventional carriers such as plasticizers, pigments, dyes, glidants, stabilizing agents, pore formers and surfactants.
[434] "Diluents", also referred to as "fillers", are typically required to increase the volume of a solid dosage form so that a convenient size is provided for tablet compression or spherule or granule formation. Suitable diluents include, but are not limited to, dicalcium phosphate dihydrate, calcium sulfate, lactose, sucrose, mannitol, sorbitol, cellulose, microcrystalline cellulose, kaolin, sodium chloride, dry starch, hydrolyzed starches, pregelatinized starch, of silicone, aluminum and magnesium silicate and powdered sugar.
[435] "Binders" are used to impart cohesive qualities to a solid dosage formulation and thus ensure that a tablet or bead or granule remains intact after formation of the dosage forms. Suitable binding materials include, but are not limited to, starch, pregelatinized starch, gelatin, sugars (including sucrose, glucose, dextrose, lactose and sorbitol), polyethylene glycol, waxes, natural and synthetic gums such as acacia, tragacanth , sodium alginate, cellulose, including hydroxypropylmethylcellulose, hydroxypropylcellulose, ethylcellulose, and veegum, and synthetic polymers, such as acrylic acid and methacrylic acid copolymers, methacrylic acid copolymers, methyl methacrylate copolymers, aminoalkyl methacrylate copolymers, polyacrylic acid /polymethacrylic acid and polyvinylpyrrolidone.
[436] "Lubricants" are used to facilitate tabletting. Examples of suitable lubricants include, but are not limited to, magnesium stearate, calcium stearate, stearic acid, glycerol behenate, polyethylene glycol, talc and mineral oil.
[437] "Disintegrants" are used to facilitate disintegration or "disintegration" of the dosage form after administration and generally include, but are not limited to, starch, sodium starch glycollate, sodium carboxymethyl starch, sodium carboxymethyl cellulose, hydroxypropyl cellulose, starch pregelatinized, alginine, gums or cross-linked polymers such as cross-linked PVP (POLYPLASDONE® XL from GAF Chemical Corp).
[438] "Stabilizers" are used to inhibit or delay drug decomposition reactions, which include, by way of example, oxidative reactions. Suitable stabilizers include, but are not limited to, antioxidants, butylated hydroxytoluene (BHT); ascorbic acid, salts and esters thereof; vitamin E, tocopherol and salts thereof; sulfites, such as sodium metabisulfite; cysteine and derivatives thereof; Citric acid; propyl, and butylated hydroxyanisole (BHA). (a) Controlled-release enteral formulations
[439]Oral dosage forms, such as capsules, tablets, solutions and suspensions, may be formulated for controlled release, for example, for the controlled release of one or more compounds as defined in section III.A above for use in according to any one of the first, second or third aspects of the present invention. For example, the one or more compounds and one or more optional additional active agents can be formulated into nanoparticles, microparticles and combinations thereof and encapsulated in a soft or hard or non-gelatinous gelatin capsule or dispersed in a dispersion medium to form a oral suspension or syrup. The particles can be formed by the active agent and a polymer or controlled release matrix. Alternatively, the active agent particles can be coated with one or more controlled release coatings prior to incorporation into the finished dosage form.
[440] In another embodiment, the one or more compounds and optional one or more additional active agents are dispersed in a matrix material, which gels or emulsifies on contact with an aqueous medium, such as physiological fluids. In the case of gels, the matrix swells with active agents, which are slowly released over time by diffusion and/or degradation of the matrix material. Such matrices can be formulated as tablets or as fillers for hard and soft capsules.
[441] In yet another embodiment, the one or more compounds, and one or more optional additional active agents are formulated into a sold oral dosage form, such as a tablet or capsule, and the solid dosage form is coated with one or more more controlled release coatings such as delayed release coatings or extended release coatings. The coating or coatings may also contain the additional active compounds and/or agents. (1) Extended-release dosage forms
[442]Sustained release formulations are generally prepared as diffusion or osmotic systems, which are known in the art. A diffusion system generally consists of two types of devices, a reservoir and a matrix, and is well known and described in the art. Matrix devices are generally prepared by compressing the drug with a slowly dissolving polymer carrier into a tablet form. The three main types of materials used in the preparation of matrix devices are insoluble plastics, hydrophilic polymers and fatty compounds. Plastic matrices include, but are not limited to, methyl acrylate-methyl methacrylate, polyvinyl chloride, and polyethylene. Hydrophilic polymers include, but are not limited to, cellulosic polymers such as methyl and ethyl cellulose, hydroxyalkyl celluloses such as hydroxypropyl cellulose, hydroxypropyl methyl cellulose, sodium carboxymethyl cellulose and CARBOPOL® 934 (cross-linked polyacrylate polymer), polyethylene oxides and mixtures thereof. Fatty compounds include, but are not limited to, various waxes, such as carnauba wax and glyceryl tristearate, and wax-like substances including hydrogenated castor oil or hydrogenated vegetable oil, or mixtures thereof.
[443] In certain preferred embodiments, the plastic material is a pharmaceutically acceptable acrylic polymer, including, but not limited to, copolymers of acrylic acid and methacrylic acid, methyl methacrylate, copolymers of methyl methacrylate, ethoxyethyl methacrylates, cyanoethyl methacrylate, aminoalkyl methacrylate, methacrylate copolymer, poly(acrylic acid), poly(methacrylic acid), methacrylic acid alkylamine copolymer, poly(methyl methacrylate), poly(methacrylic acid)(anhydride), polymethacrylate, polyacrylamide, poly(methacrylic acid anhydride) and glycidyl methacrylate copolymers.
[444] In certain preferred embodiments, the acrylic polymer is comprised of one or more ammonium methacrylate copolymers. Ammonium methacrylate copolymers are well known in the art and are described in NF XVII as fully polymerized copolymers of acrylic and methacrylic acid esters with a low content of quaternary ammonium groups.
[445] In a preferred embodiment, the acrylic polymer is an acrylic resin varnish, such as is commercially available from Rohm Pharma under the tradename EUDRAGIT®. In other preferred embodiments, the acrylic polymer comprises a blend of two acrylic resin varnishes commercially available from Rohm Pharma under the tradenames EUDRAGIT® RL30D and EUDRAGIT® RS30D, respectively. EUDRAGIT® RL30D and EUDRAGIT® RS30D are copolymers of acrylic and methacrylic esters with a low content of quaternary ammonium groups, the molar ratio of ammonium groups to the remaining neutral (meth)acrylic esters being 1:20 in EUDRAGIT® RL30D and 1 :40 on EUDRAGIT® RS30D. The average molecular weight is about 150,000. EUDRAGIT ® S-100 and EUDRAGIT ® L-100 are also preferred. The code designations RL (high permeability) and RS (low permeability) refer to the permeability properties of these agents. EUDRAGIT® RL/RS blends are insoluble in water and digestive fluids. However, the multiparticulate systems formed to include the same are swellable and permeable in aqueous solutions and digestive fluids.
[446] The polymers described above, such as EUDRAGIT® RL/RS, can be blended together at any desired ratio to ultimately obtain a sustained release formulation with a desirable dissolution profile. Desirable sustained release multiparticulate systems can be obtained, for example, from 100% EUDRAGIT® RL, 50% EUDRAGIT® RL and 50% EUDRAGIT t® RS and 10% EUDRAGIT® RL and 90% EUDRAGIT ® RS. One skilled in the art will recognize that other acrylic polymers may also be used, such as, for example, EUDRAGIT® L.
[447]Alternatively, sustained release formulations can be prepared using osmotic systems or by applying a semipermeable coating to the dosage form. In the latter case, the desired drug release profile can be obtained by combining low permeability and high permeability coating materials in suitable proportions.
[448] The devices with different drug delivery mechanisms described above can be combined into a final dosage form containing single or multiple units. Examples of multiple units include, but are not limited to, multilayer tablets and capsules containing tablets or beads or granules. An immediate-release portion may be added to the extended-release system by applying an immediate-release layer over the extended-release core using a coating or compression process, or in a multiple unit system such as a capsule containing beads of prolonged and immediate release.
[449] Extended-release tablets containing hydrophilic polymers are prepared by techniques commonly known in the art, such as direct compression, wet granulation or dry granulation. Their formulations usually incorporate polymers, diluents, binders, and lubricants as well as the active pharmaceutical ingredient. Usual diluents include inert powdery substances such as starches, powdered cellulose, especially crystalline and microcrystalline cellulose, sugars such as fructose, mannitol and sucrose, cereal flours and similar edible powders. Typical diluents include, for example, various types of starch, mannitol, kaolin, calcium phosphate or sulfate, lactose, inorganic salts such as sodium chloride and powdered sugar. Powdered cellulose derivatives are also useful. Typical tablet binders include substances such as starch, gelatin and sugars such as lactose, fructose, and glucose. Natural and synthetic gums including acacia, alginates, methylcellulose and polyvinylpyrrolidone can also be used. Polyethylene glycol, hydrophilic polymers, ethyl cellulose and waxes can also serve as binders. A lubricant is needed in a tablet formulation to prevent the tablet and punches from adhering to the die. The lubricant is chosen from slippery solids such as talc, magnesium and calcium stearate, stearic acid and hydrogenated vegetable oils.
[450] Extended-release tablets containing wax materials are generally prepared using methods known in the art, such as a direct mixing method, a freezing method, and an aqueous dispersion method. In the freezing method, the drug is mixed with a waxy material and spray-frozen or frozen and sorted and processed. delayed release dosage forms
[451]Delayed release formulations can be created by coating a solid dosage form with a polymer film, which is insoluble in the acidic environment of the stomach, and soluble in the neutral environment of the small intestine.
[452]Delayed release dosage units can be prepared, for example, by coating an active agent or a composition containing an active agent with a selected coating material. The composition containing the active agent can be, for example, a tablet for incorporation into a capsule, a tablet for use as an inner core in a "coated core" dosage form, or a plurality of spheres, particles or granules containing the active agent. active agent, for incorporation into a tablet or capsule. Preferred coating materials include biodegradable, gradually hydrolysable, gradually water-soluble and/or enzymatically degradable polymers, and may be conventional "enteric" polymers. Enteric polymers, as will be appreciated by those skilled in the art, become soluble in the higher pH environment of the lower gastrointestinal tract or slowly erode as the dosage form passes through the gastrointestinal tract, whereas enzymatically degradable polymers are degraded by bacterial enzymes present in the lower gastrointestinal tract, particularly in the colon. Suitable coating materials to effect delayed release include, but are not limited to, cellulosic polymers such as hydroxypropyl, hydroxyethyl cellulose, hydroxymethyl cellulose, hydroxypropyl methyl cellulose, hydroxypropyl methyl cellulose acetate succinate, hydroxypropyl methyl cellulose phthalate, methyl cellulose, ethyl cellulose, cellulose acetate , cellulose acetate phthalate, cellulose acetate trimellitate and sodium carboxymethyl cellulose; acrylic acid polymers and plastics, preferably formed from acrylic acid, methacrylic acid, methyl acrylate, ethyl acrylate, methyl methacrylate and/or ethyl methacrylate and other methacrylic resins commercially available under the trade name EUDRAGIT® (Rohm Pharma; Westerstadt, Germany), including EUDRAGIT® L30D-55 and L100-55 (soluble at pH 5.5 and above), EUDRAGIT® L-1OO (soluble at pH 6.0 and above), EUDRAGIT® S (soluble in pH 7.0 and higher, as a result of a higher degree of esterification) and EUDRAGITS® NE, RL and RS (water-insoluble polymers with different degrees of permeability and expandability); vinyl polymers and copolymers such as polyvinyl pyrrolidone, vinyl acetate, acetate phthalate, crotonic acid acetate copolymer and ethylene-vinyl acetate copolymer; enzymatically degradable polymers such as azo polymers, pectin, chitosan, amylose and guar gum; zein and shellac. Combinations of different coating materials can also be used. Multilayer coatings using different polymers can also be applied.
[453] Preferred coating weights for particular coating materials can be readily determined by those skilled in the art by evaluating individual release profiles for tablets, spheres and granules prepared with different amounts of various coating materials. It is the combination of materials, method and form of application that produce the desired release characteristics, which can only be determined from clinical studies.
[454]The coating composition may include conventional additives such as plasticizers, pigments, dyes, stabilizing agents, glidants, etc. A plasticizer is normally present to reduce the brittleness of the coating, and generally accounts for about 3 to 50% by weight. or 10% by weight to 50% by weight, based on the dry weight of the polymer. Examples of typical plasticizers include polyethylene glycol, propylene glycol, triacetin, dimethyl phthalate, diethyl phthalate, dibutyl phthalate, dibutyl sebacate, triethyl citrate, tributyl citrate, triethylacetyl citrate, castor oil and acetylated monoglycerides. A stabilizing agent is preferably used to stabilize the dispersed particles. Typical stabilizing agents are non-ionic emulsifiers such as sorbitan esters, polysorbates and polyvinylpyrrolidone. Sliding agents are recommended to reduce adhesion effects during film formation and drying, and generally account for approximately 25% by weight to 100% by weight. of the weight of the polymer in the coating solution. An effective gliding agent is talc. Other glidants, such as magnesium stearate and glycerol monostearates, may also be used. Pigments such as titanium dioxide may also be used. Small amounts of an anti-foaming agent such as a silicone (e.g. simethicone) may also be added to the coating composition. 3. Topical formulations
[455] Compounds as defined in section III.A above for use in accordance with any of the first, second or third aspects of the present invention and may be formulated for topical administration.
[456]The formulations may contain one or more compounds discussed above, alone or in combination, in an amount effective to prevent or inhibit biofilm formation on a surface, or reduce the amount of biofilm on a surface to be treated. 1000 colony forming units (cfu) of Campylobacter is enough to infect a human and cause disease in a human.
[457] Accordingly, in one embodiment, an effective amount of the one or more compounds as defined in section III.A of this application is sufficient of the compound(s), alone or in combination with other compounds, to reduce the number cfu of Campylobacter or other microorganism of interest on the surface to be treated to a number that is unlikely or will not cause infection in humans.
[458] Dosage forms suitable for topical administration include creams, ointments, balms, sprays, gels, lotions, irrigants, and emulsions.
[459] "Buffers" are used to control the pH of a composition. Preferably, the buffers buffer the composition from a pH of about 4 to a pH of about 7.5, more preferably from a pH of about 4 to a pH of about 7, and most preferably from a pH of about 7. a pH of about 5 to a pH of about 7. In a preferred embodiment, the buffer is triethanolamine.
[460] "Emollients" are an externally applied agent that softens or smoothes the skin and are generally known in the art and listed in textbooks such as the "Handbook of Pharmaceutical Excipients", 4th Ed., Pharmaceutical Press, 2003. These include, without limitation, almond oil, castor oil, keratonium extract, cetostearoyl alcohol, cetyl alcohol, cetyl esters wax, cholesterol, cottonseed oil, cyclomethicone, ethylene glycol palmitostearate, glycerin, glycerin monostearate, glyceryl monooleate, isopropyl myristate, isopropyl palmitate, lanolin, lecithin, light mineral oil, medium chain triglycerides, mineral oil and lanolin alcohols, petrolatum, petrolatum and lanolin alcohols, soybean oil, starch, stearyl alcohol, sunflower oil, xylitol and combinations thereof. In one embodiment, the emollients are ethylhexyl stearate and ethylhexyl palmitate.
[461] "Emulsifiers" are surface-active substances that suspend one liquid in another and promote the formation of a stable mixture, or emulsion, of oil and water. Common emulsifiers are: metallic soaps, certain animal and vegetable oils, and various polar compounds. Suitable emulsifiers include gum arabic, anionic emulsifying wax, calcium stearate, carbomers, cetostearyl alcohol, cetyl alcohol, cholesterol, diethanolamine, ethylene glycol palmitostearate, glycerin monostearate, glyceryl monooleate, hydroxypropyl cellulose, hypromellose, lanolin, alcohols lanolin, hydrate, lecithin, medium chain triglycerides, methyl cellulose alcohols, mineral oil and lanolin, monobasic sodium phosphate, monoethanolamine, non-ionic emulsifying wax, oleic acid, poloxamer, poloxamers, polyoxyethylene alkyl ethers, castor oil derivatives polyoxyethylene, polyoxyethylene sorbitan fatty acid esters, polyoxyethylene stearates, propylene glycol alginate, glyceryl monostearate self-emulsifiers, sodium citrate dihydrate, sodium lauryl sulfate, sorbitan esters, stearic acid, sunflower oil, tragacanth, triethanolamine, xanthan gum and combinations thereof. In one embodiment, the emulsifying agent is glycerol stearate.
[462] "Penetration enhancers" are known in the art and include, but are not limited to, fatty alcohols, fatty acid esters, fatty acids, fatty alcohol ethers, amino acids, phospholipids, lecithins, cholate salts, enzymes, amines and amides, complexing agents (liposomes, cyclodextrins, modified celluloses, and diimides), macrocyclics such as macrocyclic lactones, ketones, and cyclic anhydrides and ureas, surfactants, N-methyl pyrrolidones and their derivatives, DMSO and related compounds, ionic compounds , azone and related compounds, and solvents such as alcohols, ketones, amides, polyols (e.g. glycols). Examples of these classes are known in the art.
[463] "Preservatives" can be used to prevent the growth of fungi and microorganisms. Suitable antifungal and antimicrobial agents include, but are not limited to, benzoic acid, butylparaben, ethyl paraben, methylparaben, propylparaben, sodium benzoate, sodium propionate, benzalkonium chloride, benzethonium chloride, benzyl alcohol, cetylpyridinium chloride, chlorobutanol , phenol, phenylethyl alcohol and thimerosal.
[464] "Surface agents" are surface agents that reduce the surface tension and thereby increase the emulsifying, foaming, dispersing, spreading and wetting properties of a product. Suitable nonionic surfactants include emulsifying wax, glyceryl monooleate, polyoxyethylene alkyl ethers, polyoxyethylene castor oil derivatives, polysorbate, sorbitan esters, benzyl alcohol, benzyl benzoate, cyclodextrins, glycerin monostearate, poloxamer, povidone and combinations thereof. In one embodiment, the nonionic surfactant is stearyl alcohol. (a) Emulsions
[465] An emulsion is a preparation of a liquid distributed in small globules throughout the body of a second liquid. In particular embodiments, the immiscible components of the emulsion include a lipophilic component and an aqueous component. The dispersed liquid is the discontinuous phase, and the dispersion medium is the continuous phase. When oil is the dispersed liquid and an aqueous solution is the continuous phase, it is known as an oil-in-water emulsion, whereas when water or aqueous solution is the dispersed phase and oil or oleaginous substance is the continuous phase, it is known as a water-in-oil emulsion. One or both of the oil and water phases may contain one or more surfactants, emulsifiers, emulsion stabilizers, buffers and other excipients. Preferred excipients include surfactants, especially nonionic surfactants; emulsifying agents, especially emulsifying waxes; and non-volatile liquid non-aqueous materials, particularly glycols, such as propylene glycol. The oil phase may contain other pharmaceutically approved oily excipients. For example, materials such as hydroxylated castor oil or sesame oil can be used in the oil phase as surfactants or emulsifiers.
[466] The oil phase may comprise, at least in part, a propellant, such as an HFA propellant. One or both of the oil and water phases may contain one or more surfactants, emulsifiers, emulsion stabilizers, buffers and other excipients. Preferred excipients include surfactants, especially nonionic surfactants; emulsifying agents, especially emulsifying waxes; and non-volatile liquid non-aqueous materials, particularly glycols, such as propylene glycol. The oil phase may contain other pharmaceutically approved oily excipients. For example, materials such as hydroxylated castor oil or sesame oil can be used in the oil phase as surfactants or emulsifiers.
[467] A subset of emulsions are self-emulsifying systems. These delivery systems are typically capsules (hard shell or soft shell) consisting of the compound dispersed or dissolved in a mixture of surfactant(s) and lipophilic liquids, such as oils or other water-immiscible liquids. When the capsule is exposed to an aqueous environment and the outer gelatin coating dissolves, contact between the aqueous medium and the capsule contents instantly generates very small emulsion droplets. These are typically in the size range of micelles or nanoparticles. No mixing force is required to generate the emulsion as is typically the case in emulsion formulation processes. (b) Lotions
[468] A lotion may contain finely powdered substances that are insoluble in the dispersion medium through the use of suspending agents and dispersing agents. Alternatively, lotions may have dispersed-phase liquid substances that are immiscible with the carrier and are usually dispersed by means of emulsifying agents or other suitable stabilizers. In one embodiment, the lotion is in the form of an emulsion having a viscosity between 100 and 1000 centistokes. The fluidity of lotions allows for quick and uniform application over a wide area. Lotions are normally meant to dry on the skin leaving a thin layer of their medicinal components on the surface of the skin. (c) creams
[469] Creams may contain emulsifying agents and/or other stabilizing agents. In one embodiment, the formulation is in the form of a cream having a viscosity greater than 1000 centistokes, typically in the range of 20,000-50,000 centistokes. Creams are often preferred over ointments, as they are generally easier to spread and easier to remove.
[470] The difference between a cream and a lotion is viscosity, which is dependent on the amount/use of various oils and the percentage of water used to prepare the formulations. Creams are typically thicker than lotions, can have multiple uses, and often use more varied oils/butters depending on the desired effect on the skin. In a cream formulation, the percentage of water base is about 60-75% and the oil base is about 20-30% of the total, with the other percentages being the emulsifying agent, preservatives and additives for a 100% total. (d) ointments
[471] Examples of suitable ointment bases include hydrocarbon bases (e.g. petrolatum, white petroleum jelly, yellow ointment and mineral oil); absorption bases (hydrophilic petrolatum, anhydrous lanolin, lanolin and cold cream); water-removable bases (eg hydrophilic ointment) and water-soluble bases (eg polyethylene glycol ointments). Pastes typically differ from ointments in that they contain a higher percentage of solids. Pastes are usually more absorbent and less greasy than ointments made from the same ingredients. (e) Gels
[472] Gels are semi-solid systems containing dispersions of small or large molecules in a liquid vehicle that is made semi-solid by the action of a thickening agent or polymeric material dissolved or suspended in the liquid vehicle. The liquid may include a lipophilic component, an aqueous component, or both. Some emulsion gels may be or include a gel component. Some gels, however, are not emulsions because they do not contain a homogenized mixture of immiscible components. Suitable gelling agents include, but are not limited to, modified celluloses, such as hydroxypropyl cellulose and hydroxyethyl cellulose; Carbopol homopolymers and copolymers; and combinations thereof. Suitable solvents in the liquid vehicle include, but are not limited to, diglycol monoethyl ether; alkylene glycols such as propylene glycol; dimethyl isosorbide; alcohols such as isopropyl alcohol and ethanol. Solvents are typically selected for their ability to dissolve the compound. Other additives may also be incorporated, which improve the skin feel and/or the emollience of the formulation. Examples of such additives include, but are not limited to, isopropyl myristate, ethyl acetate, C12-C15 alkyl benzoates, mineral oil, squalane, cyclomethicone, capric/caprylic triglycerides and combinations thereof. (f) Foams
[473] Foams consist of an emulsion in combination with a gaseous propellant. The gaseous propellant mainly consists of hydrofluoroalkanes (HFA). Suitable propellants include HFAs such as 1,1,1,2-tetrafluoroethane (HFA 134a) and 1,1,1,2,3,3,3-heptafluoropropane (HFA 227), but mixtures thereof and other HFAs that are approved or may be approved for medical use are suitable. Preferably, the propellants are not hydrocarbon propellant gases, which can produce flammable or explosive vapors during spraying. Furthermore, the compositions preferably do not contain volatile alcohols, which can produce flammable or explosive vapors during use. 4. Disinfection and cleaning formulations
[474] Compounds as defined in section III.A above for use in accordance with any one of the first, second or third aspects of the present invention may be formulated into cleansing formulations.
[475]Cleaning formulations include formulations that are highly effective for household cleaning applications (e.g. hard surfaces such as floors, countertops, sinks, tiles, tableware and softer cloth materials such as clothes, sponges, paper towels, etc.), personal care applications (e.g. lotions, shower gels, soaps, shampoos, sprays, wipes, toothpaste, acne treatments, skin cleansers, mouthwash, wound irrigation solutions, towels, contact lenses and lens housings) and industrial and hospital applications (e.g. antifouling coatings and disinfection of instruments, medical devices, gloves, filters, membranes, tubes, drains, piping including gas piping, oil piping, drill pipes, frac pipes, sewer pipes, drainage pipes, hoses, animal carcasses, aquariums, showers, children's toys, boat hulls, and cooling towers). These formulations are effective for cleaning surfaces that are infected or contaminated with biofilm or for preventing biofilm formation on these surfaces.
[476] The compounds may be formulated in a solution in a suitable solvent for administration in a spray bottle, the compounds may be formulated as an aerosol, as a foam, suitable for spraying onto surfaces, or, they may be soaked in a cloth or other item suitable for cleaning a surface to be disinfected. Methods of making formulations for use as a disinfectant on molds are known in the art.
[477] One embodiment provides the compounds, or a derivative thereof, in a composition that contains a pH-indicating dye and an alkaline substance. The pH indicator dye indicates that the surface has been disinfected and ensures that sufficient time has passed to disinfect the surface. See, for example, US Patent 20140057987, which is incorporated herein by reference in its entirety.
[478]Cleansing formulations may include the compounds and an acceptable carrier. Carrier can be in a wide variety of forms. For example, the carrier can be an aqueous solution or cleaning agent, an alcohol-based solution or gel, or an emulsion carrier, including but not limited to oil-in-water, water-in-oil, water -in-water, oil-in-water, and oil-in-water-in-silicone emulsions. The carrier solution containing the compound(s) can be applied directly to the surface to be treated or delivered through a suitable substrate.
[479]Cleansing formulations can be formulated for use on the skin. In these embodiments the compounds may be formulated in a dermatologically acceptable carrier. Dermatologically acceptable carriers can also be, for example, formulated as alcohol- or water-based hand cleansers, bath bars, liquid soaps, shampoos, shower gels, hair conditioners, hair tonics, pastes or mousses. .
[480]Cleaning formulations may contain one or more surfactants. The surfactant is suitably selected from anionic, nonionic, zwitterionic, amphoteric and ampholytic surfactants, as well as mixtures of these surfactants. Such surfactants are well known to those skilled in the detergency art. Non-limiting examples of possible surfactants include isocetet-20, sodium methyl coccotyl taurate, sodium methyl oleyl taurate and sodium lauryl sulfate. Examples of a wide variety of additional surfactants are described in McCutcheon's Detergents and Emulsifiers. North American Edition (1986), published by Allured Publishing Corporation. Cleaning formulations may optionally contain, at their art-established levels, other materials that are conventionally used in cleaning formulations.
[481] Additional carriers suitable for cleaning formulations may include various substrate-based products. In such cases, the present formulations may be impregnated in or on the substrate products and may be allowed to remain wet or may be subjected to a drying process. For example, suitable vehicles include, but are not limited to, wet and dry wipes suitable for personal care and home use (e.g. non-woven baby wipes, household cleaning wipes, surgical prep wipes, etc.) ; diapers; children's pillows; floss; personal care and home care sponges or woven cloths (eg wipes, towels, etc.); tissue-type products (eg facial tissue, paper towels, etc.); and disposable clothing (eg gloves, blouses, surgical masks, children's bibs, socks, shoe inserts, etc.). Cleaning formulations can be incorporated into various home care products including, but not limited to, hard surface cleaners (eg, disinfectant sprays, liquids or powders); dish detergents or detergents (liquid or solid), floor waxes, glass cleaners, etc.
[482] Examples of carriers may include aqueous solutions, for example, having from about 0% to about 98.8%, by weight of the composition, of water. In addition, the carriers may contain an aqueous alcohol solution. The amount of alcohol present in the alcohol solution will vary depending on the type of product into which the composition is incorporated, i.e. a cleaning in which the preferred amount of alcohol present would be from about 0% to about 25%, while a hand sanitizer preferably contains from about 60% to about 95% alcohol. Accordingly, suitable dermatologically acceptable alcohol solutions or gels may contain from about 0% to about 95%, by weight of the composition, of an alcohol.
[483] Alcohols suitable for inclusion in alcohol solutions of the vehicle include, but are not limited to, monohydric alcohols, dihydric alcohols and combinations thereof. The most preferred alcohols are selected from the group consisting of C2-C18 linear or branched monohydric alcohols. More preferred alcohols are selected from the group consisting of ethanol, isopropanol, n-propanol, butanol, and combinations thereof. Cleaning formulations that contain an alcohol solution may contain anhydrous or water.
[484] Thickeners can be added to water or alcohol based to form a gel. Examples of suitable thickeners include, but are not limited to, naturally occurring polymeric materials such as sodium alginate, xanthan gum, quince seed extract, gum tragacanth, starch and the like, semi-synthetic polymeric materials such as ethers of cellulose (e.g. hydroxyethylcellulose, methylcellulose, carboxymethylcellulose, hydroxypropylmethylcellulose), polyvinylpyrrolidone, polyvinyl alcohol, guar gum, hydroxypropyl guar gum, soluble starch, cationic celluloses, cationic guars and the like, and synthetic polymeric materials such as carboxyvinyl polymers, polyvinylpyrrolidone, alcohol polyvinyl, polyacrylic acid polymers, polymethacrylic acid polymers, polyvinyl acetate polymers, polyvinyl chloride polymers and polyvinylidene chloride polymers. Inorganic thickening agents may also be used, such as aluminum silicates, such as, for example, bentonites, or a mixture of polyethylene glycol and polyethylene glycol stearate or distearate.
[485]Cleansing formulations may contain, in addition to the compounds described above, one or more antimicrobial or antifungal agents. Such agents are capable of destroying microbes, preventing the development of microbes or preventing the pathogenic action of microbes. Examples of additional antimicrobial and antifungal agents include β-lactam drugs, quinolone drugs, ciprofloxacin, norfloxacin, tetracycline, erythromycin, amikacin, 2,4,4'-trichloro-2'-hydroxydiphenyl ether (TRICLOSAN®), phenoxyethanol, phenoxypropanol , phenoxysopropanol, doxycycline, capreomycin, chlorhexidine, chlortetracycline, oxytetracycline, clindamycin, ethambutol, hexamidine isethionate, metronidazole, pentamidine, gentamicin, kanamycin, lineomycin, methacycline, methenamine, minocycline, neomycin, streptomycin, tobramycin, miconazole, tetratromycin hydrochloride, ether , zinc erythromycin, erythromycin estolate, erythromycin stearate, amikacin sulfate, doxycycline hydrochloride, capreomycin sulfate, chlorhexidine hydrochloride, chlorethracycline hydrochloride, oxytetracycline hydrochloride, clindamycin hydrochloride, ethambutol hydrochloride, metronidazole hydrochloride, metronidazole hydrochloride pentamidine, gentamicin sulfate, kanamycin sulfate, lineomycin hydrochloride, methacycline hydrochloride, methenamine hippurate, methenamine mandelate, minocycline hydrochloride, neomycin sulfate, netilmycin sulfate, paromomycin sulfate, streptomycin sulfate, tobramycin sulfate, miconazole hydrochloride, ketaconazole, amanfadine hydrochloride, amanfadine sulfate, octopirox, parachlorometa xylenol, nystatin, tolnaphthate, pyrithiones (especially zinc pyrithione, also known as ZPT), dimethyldimethylol hydantoin (GLYDANT®), methylchloroisothiazolinone/methylisothiazolinone (KATHON CG®), sodium sulfite, sodium bisulfite sodium, imidazolidinyl urea (Germall 115®), diazolidinyl urea (GERMAILL II®), benzyl alcohol, 2-bromo-2-nitropropane-1,3-diol (BRONOPOL®), formalin (formaldehyde), iodopropenyl butylcarbamate (POLYPHASE P100) ®) chloroacetamide, methanamine, methyldibromonitril glutaronitrile (1,2-dibromo-2,4-dicyanobutane or TEKTAMER®), glutaraldehyde, 5-bromo-5-nitro-1,3-dioxane (BRONIDOX®), alcohol f ethyl, sodium o-phenylphenol/o-phenylphenol, sodium hydroxymethylglycinate (SUTTOCIDE A®), polymethoxybicyclic oxazolidine (NUOSEPt C®), dimethoxane, dichlorobenzyl thimerosal alcohol, captan, chlorophenenosine, dichlorophene, chlorobutane, glyceryl laurate, halogenated diphenyl ethers such as 2,4,4'-trichloro-2'-hydroxy-diphenyl ether (TRICLOSAN® or TCS), 2,2'-dihydroxy-5,5'-dibromodiphenylether, phenolic compounds such as phenol, 2-methylphenol, 3 -methylphenol, 4-methylphenol, 4-ethylphenol, 2,4-dimethylphenol, 2,5-dimethylphenol, 3,4-dimethylphenol, 2,6-dimethylphenol, 4-n-propylphenol, 4-n-butylphenol, 4-n-amylphenol, 4-tert-amylphenol, 4-n-hexylphenol, 4-n-heptylphenol, mono- and polyalkyl and aromatic halophenols, such as p-chlorophenol, methyl p-chlorophenol, ethyl, n-propyl p-chlorophenol, n-butyl p-chlorophenyl, n-amyl p-chlorophenol, sec-amyl p-chlorophenol, n-hexyl p-chlorophene, cyclohexyl p-chlorophenol, n-heptyl chlorophenol, n-octyl p-chlorophenol, o-chlorophenol, methyl o-chlorophenol, ethyl o-chloro rophenol, n-propyl o-chlorophenol, n-butyl o-chlorophenol, n-amyl o-chlorophenol, tert-amyl o-chlorophenol, n-hexyl o-chlorophenol, n-heptyl o-chlorophenol, o-benzyl p-chlorophenol , o-benzyl-m-methyl p-chlorophenol, o-benzyl-m, m-dimethyl p-chlorophenol, o-phenylethyl, p-chlorophenol, o-phenylethyl-m-methyl p-chlorophenol, 3-methyl p-chlorophenol , 3,5-dimethyl p-chlorophenol, 6-ethy 1-3-methyl p-chlorophenol, 6-n-propyl-3-methyl p-chlorophenol, 6-isopropyl-3-methyl p-chlorophenol, 2-ethyl- 3,5-dimethyl p-chlorophenol, 6-sec-butyl-3-methyl p-chlorophenol, 2-isopropyl-3,5-dimethyl p-chlorophenol, 6-diethylmethyl-3-methyl-p-chlorophenol, 6-iso- propyl-2-ethyl-3-methyl p-chlorophenol, 2-sec-amyl-3,5-dimethyl p-chlorophenol, 2-diethylmethyl-3,5-dimethyl p-chlorophenol, 6-sec-octyl-3-methyl p-chlorophenol, p-chloro-m-cresol, p-bromophenol, methyl p-bromophenol, ethyl p-bromophenol, n-propyl p-bromophenol, n-butyl p-bromophenol, n-amyl p-bromophenol, sec-amyl p-bromophenol, n-hexyl p-bromophenol, cyclohexyl p-bromophenol, o-bromophenol, tert-amyl o-bromophenol, n-hex yl o-bromophenol, n-propyl-m,m-dimethyl o-bromophenol, 2-phenyl phenol, 4-chloro-2-methyl phenol, 4-chloro-3-methyl phenol, 4-chloro-3,5-dimethyl phenol, 2,4-dichloro-3,5-dimethylphenol, 3,4,5,6-terabromo-2-methylphenol, 5-methyl-2-pentylphenol, 4-isopropyl-3-methylphenol, para-chloro-meta- xylenol (PCMX), chlorothymol, 5-chloro-2-hydroxydiphenylmethane, resorcinol and derivatives thereof including methyl resorcinol, ethyl resorcinol, n-propyl resorcinol, n-butyl resorcinol, n-amyl resorcinol, n-hexyl resorcinol, n-heptyl resorcinol , n-octyl resorcinol, n-nonyl resorcinol, phenyl resorcinol, benzyl resorcinol, phenylethyl resorcinol, phenylpropyl resorcinol, p-chlorobenzyl resorcinol, 5-chloro 2,4-dihydroxydiphenyl methane, 4'-chloro 2,4-dihydroxydiphenyl methane , 5-bromo 2,4-dihydroxydiphenyl methane, and 4'-bromo 2,4-dihydroxydiphenyl methane, biphenolic compounds 2,2'-methylene bis(4-chlorophenol), 2,2'-methylene bis(3, 4,6-trichlorophenol), 2,2'-methylene bis(4-chloro-6-bromophenol), bis(2-hydroxy-3,5-dichlorophenyl) sulfide, and bis(2-hydroxy-5-chlorobenzyl)sulfide, benzoic esters (parabens) such as methylparaben, propylparaben, butylparaben, ethylparaben, isopropylparaben, isobutylparaben, benzylparaben, sodium methylparaben and sodium propylparaben, halogenated carbanilides (e.g. 3,4, 4'-Trichlorocarbanilides (TRICLOCARBAN® or TCC), 3-trifluoromethyl-4,4'-dichlorocarbanilide, 3,3',4-trichlorocarbanilide, etc.), cationic actives such as benzalkonium chloride and clotrimazole. Another class of antimicrobial agents (specifically antibacterial agents) that are useful are so-called "natural" antibacterial actives, referred to as natural essential oils. Typical natural essential oil antibacterial actives include anise, lemon, orange, rosemary, wintergreen, thyme, lavender, clove, hop, tea tree, citronella, wheat, barley, lemongrass, cedar, cedar, cinnamon, fleagrass, geranium, sandalwood, violet, cranberry, eucalyptus, vervain, peppermint, benzoin gum, basil, fennel, fir, balm, menthol, ocmea origanum, Hydastis carradensis, Berberidaceae daceae, Ratanhiae and Curcuma longa.
[486] The cleaning formulations may be packaged in a variety of suitable packages known to those skilled in the art. Liquid formulations may desirably be packaged in hand-operated spray dispensing containers, which are usually made of synthetic organic polymeric plastic materials. Accordingly, disinfectant formulations containing the compounds and packaged in a spray dispenser, preferably a trigger spray dispenser or a pump spray dispenser, are envisaged. Spray-type dispensers allow the liquid cleaning formulations described herein to be uniformly applied to a relatively large area of a surface to be disinfected.
[487] The compounds can be soaked in a non-woven absorbent cloth. Disinfectant wet wipes are also described, for example, in US Patent 8,563,017.
[488] The compounds may be in the form of an aqueous foam with a special surfactant system capable of generating a foam. See US Patent 8,097,265, US Patent 5,891,922 and US Patent 4,889,645.
[489] The compounds may also be in a pressurized aerosol spray. See also US 20010053333 which describes an instant-drying liquid aerosol disinfectant composition with a flash spray component and an effective amount of an antimicrobial agent.
[490] It is within the skill of one skilled in the art to determine the effective amount of compounds to include in a disinfectant aerosol, foam, solution or cloth for the purpose of sterilizing, for example, high-risk hospital surfaces. D. Conjugation and immobilization of compounds
[491] The one or more compounds as defined in section III.A above for use in accordance with any of the first, second or third aspects of the present invention and may be presented as conjugated and/or immobilized compounds.
[492] Compounds can be conjugated with other agents in order to retain the compounds on surfaces, for example to prevent biofilm formation on a surface. In one embodiment, the compounds may be conjugated to an agent that has an affinity for a surface in order to retain the compounds on that surface. For example, the compounds can be conjugated to an agent where the agent is a polymer or oligomer, and the polymer or oligomer has a high affinity for the surface.
[493] In another embodiment, the compounds can be conjugated to an agent wherein the agent comprises a reactive moiety suitable for anchoring to a surface. The reactive fraction may, for example, be photoreactive, or capable of covalently coupling to a surface. The reactive moiety may also incorporate spacers and linkers and other functional groups in order to place the compound in a desired location relative to the surface. Figures 15A-C are examples of how FeQ (Fe-QA) can be conjugated to an agent comprising a reactive moiety suitable for anchoring to a surface. In each of the three examples, FeQ is conjugated to a calix [4] arene framework that comprises a reactive group. In Figure 15A, FeQ is conjugated via a ligand to a calix [4] arene framework that contains a photoreactive functional group. Figure 15B is a variant of Figure 15A showing that the reactive fraction can be positioned at a different site in the calix [4] arene structure. Figure 15C is an example of FeQ conjugated to a calix [4] arene structure, where the latter is functionalized with thiol groups that are capable of reacting with surfaces. It should be understood that different linkers or non-linkers may be used and that other agents may be used in place of the calix[4]arene structure, including cyclodextrins and other polymers and oligomers.
[494] In yet another embodiment, the compounds may be conjugated to an agent comprising a substance with an affinity for a surface. The agent may incorporate spacers and linkers and other functional groups so as to place the compound in a desired location relative to the surface. In one embodiment, the agent contains hydroxyapatite. Figures 16A and B are examples of how FeQ (Fe-QA) can be conjugated via a linker to hydroxyapatite. In these examples, the linkers are attached at different positions to one of the quinic acid ligands through a functional group, Y', and at the other end of the linker they are attached to hydroxyapatite (HA) through a second functional group, X'. In an alternative embodiment, the HA group in the structures of Figures 16A and B can be used in place of a reactive group that can bond (or be bonded to) a surface, such as a photoreactive compound, isocyanate, hydroxy group, amine, ether trialkoxysilyl, such as a triethoxysilyl ether, or phosphate ester. These groups can be directly linked to the polyethylene glycol, or an additional linker inserted between the reactive group and the polyethylene glycol. E. Foods and food supplements
[495] In accordance with the first aspect of the present invention, another embodiment of the present invention provides that the compounds as defined in section III.A can be formulated into growth promoting formulations.
[496] The one or more compounds may be used, for example, in the feed or formula to improve chicken growth, for example, a meat type chicken, such as broiler, or a laying hen, such as a pullet. or a hen, or a broiler, other poultry, such as a turkey, geese, quail or ducks, or livestock, such as cattle, sheep, goats or swine, alpaca, banteng, bison, camel, cat, deer, dog, donkey, llama, mule, rabbit, reindeer, water buffalo, yak, although one skilled in the art may appreciate that other animal foods, including zoo animals, captive animals, game animals, fish (including freshwater and saltwater, farmed fish and ornamental fish), other marine and aquatic animals, including crustaceans such as oysters, mussels, shellfish, shrimp, lobster, crayfish, crabs, cuttlefish, octopus and squid, animals domestic animals such as cats and dogs, rodents (such as ngos, rats, guinea pigs, hamsters) and horses, as well as any other domestic, wild and farmed animals, including mammals, marine animals, amphibians, birds, reptiles, insects and other invertebrates. The one or more compounds can be added to drinking water for any of said animals to improve growth
[497] The compounds may be useful in the treatment of ponds, ponds or other aquatic or marine environments containing fish (including freshwater and saltwater fish, farmed fish and ornamental fish), other marine and aquatic animals including shellfish or crustaceans, such as shrimp, oysters, mussels, clams, prawns, lobsters, crayfish, crabs, cuttlefish, octopus and crayfish.
[498] The one or more compounds may be used alone or in combination with other antimicrobial, bactericidal or bacteriostatic compounds (e.g. according to the second or third aspect of the present invention) and/or growth enhancing agents.
[499] Compounds as defined in section III.A may enhance growth performance and may be used to increase average body weight during growth. The compounds can also be used to improve the feed conversion ratio. In particular, the compounds can be used to decrease mortality-adjusted feed conversion ratios (MFCR). The compounds can be used to produce animals with a higher average body weight in a given period, or they can be used to arrive at a target average body weight in a shorter period. The compounds can be used to decrease the amount of feed required for an animal to reach a target weight. In addition, the compounds can be used in stressed environments to improve growth and MFCR. These environments include, but are not limited to, high stocking densities, dirty chicken coop litter, presence of pathogens, presence of Campylobacter and other bacteria, and high temperature environments.
[500] The compositions are particularly useful in commercial poultry feeds, such as chickens, turkeys, pheasants, and ducks. Exemplary poultry foods in which one or more compounds defined in section III.A may be included include poultry foods which are referred to as "complete" foods because they are designed to contain all of the protein, energy , vitamins, minerals and other nutrients needed for proper growth, egg production and bird health. Feeding any other ingredient, mixed with the food or fed separately, changes the balance of nutrients in the "complete" food. Adding additional grains or supplementation to the complete poultry feed is not recommended.
[501] Broilers used in commercial production of optimized broilers are typically fed different diets depending on their age. For example, broiler chickens can be raised using three diets. These diets are typically called "starter", "growth" and "finish". The starter diet can be fed for about 10-12 days (typically in the 7-14 day range). This starter diet is followed by the farmer's diet, which is fed to broilers for almost 2 weeks (typically around 11-24 days). The finishing diet is used for the remainder of the production period (typically 24 to 42 days). Some broiler houses will use more or less diets (eg 4 diets), and will vary the timing of diet changes. Broilers are typically harvested between 30 and 42 days, although this time can be longer or shorter. More details and options are discussed above in the context of the first aspect of the present invention. F. Treatment to promote growth
[502] As discussed in more detail above, in the context of the first aspect of the present invention, one or more compounds defined in section III.A of this application, above, have been found to be particularly useful for promoting growth. The compounds can be added to animal feed or animal drinking water in order to promote growth. Adding the compounds to food or drinking water results in improved growth. It has also been found that the compounds can be added to animal feed or animal drinking water in order to decrease the mortality-adjusted feed conversion ratio. Thus, it is possible to use the compounds to decrease the amount of food needed for an animal to grow. The compounds can also be administered with other animal additives, and can be administered in commercial foods. In a preferred embodiment, the compounds are administered in food.
[503] It has also been found that the compounds can be administered to animals that are in a stressed environment in order to improve their growth performance. In a stressed environment the compounds promote growth that produces animals with higher average body weights. The compounds also lower mortality-adjusted feed conversion ratios in stressed environments. Examples
[504] The following non-limiting examples are included to demonstrate particular embodiments of various aspects of the present invention. It should be appreciated by those skilled in the art that the techniques disclosed in the examples which follow represent techniques discovered by the inventor to function well in the practice of the invention, and thus can be considered to constitute preferred modes for its practice. However, those skilled in the art should, in light of the present disclosure, appreciate that many changes can be made to the specific embodiments which are disclosed and still obtain a similar or similar result without departing from the spirit and scope of the invention. Example 1. Inhibition of biofilm formation on the surface of beads by Enterococcus faecalis NCTC 12697 using Fe-QA Materials and Methods
[505] Bacteria (Enterococcus faecalis NCTC 12697, Staphylococcus epidermidis F1513 and Staphylococcus aureus ATCC 25923) were cultured in brain-heart infusion (BHI) passages into new medium containing Fe-QA or isolated. Growth suspensions were prepared at 0.0001 OD/ml and then grown at 37°C under normal atmospheric conditions for 24 h in BHI with plastic coated UV beads (Lascells). After 48 h, 10 µl of suspension was serially diluted 10-fold to 10-3, 10-4, 10-5, 10-6, 10-7, 10-8. For each dilution, 10 μl were placed on BHI agar plates and colonies were counted after 24 h. The granules were also removed and washed in PBS before addition to 1 ml of PBS. After vortexing, 10 µl of the cell suspension was serially diluted as above and cell counts were performed. Results
[506] Enterococcus faecalis causes many of the antibiotic-resistant infections in hospitals, a consequence of its inherent resistance to certain antibiotics and its ability to survive and proliferate in the intestinal tract. A Ser/Thr kinase in Enterococcus faecalis has been found to mediate antimicrobial resistance. Studies have shown that PrkC, a component signaling protein containing a eukaryotic-like Ser/Thr kinase domain, enabled the inherent antimicrobial resistance and intestinal persistence of E. faecalis (Kristich, et al., Proc. Nat. Acad Sci. USA, 104(9):3508-3513 (2007)). Kristich, et al. found that an E. faecalis mutant lacking PrkC grew at a wild-type rate in the absence of antimicrobial strain, but showed greater sensitivity to active compounds involving cells, including antibiotics that targeted cell wall biogenesis and bile detergents. PrkC regulates physiological processes in E. faecalis that are critical to its success as a nosocomial pathogen.
[507] The effect of Fe-QA on biofilm formation by E. faecalis was tested as described in materials and methods. The data (Fig. 1A and 1B) show that Fe-QA inhibited biofilm formation of E. faecalis measured after treatment of E. faecalis grown on plastic coated UV beads. Example 2. Inhibition of biofilm formation on the surface of beads by Staphylococcus epidermidis F1513 using Fe-QA Materials and Methods
[508] The effect of Fe-QA on biofilm formation by S. epidermidis F1513 was tested as described in the materials and methods of Example 1. Results
[509] The data (Fig. 2A and 2B) show that Fe-QA inhibited S. epidermidis biofilm formation measured after treatment of S. epidermidis grown on plastic-coated UV beads. Example 3. Inhibition of biofilm formation on the surface of Staphylococcus aureus ATCC 25923 beads using Fe-QA
[510] The treatment of choice for S. aureus infection is penicillin; in most countries, however, penicillin resistance is extremely common, and first-line therapy is most commonly a penicillinase-resistant e-lactam antibiotic (eg, oxacillin or flucloxacillin). Combination therapy with gentamicin can be used to treat serious infections such as endocarditis, but its use is controversial because of the high risk of kidney damage (Cosgrove, et al., Clin Infect Dis, 48(6):713-721 (2009) ) Duration of treatment depends on site of infection and severity Materials and Methods
[511] The effect of Fe-QA on biofilm formation by S. epidermidis was tested as described in the materials and methods of Example 1. Results
[512] The data (Fig. 3A and 3B) show that Fe-QA inhibited S. aureus biofilm formation as measured after treatment of S. aureus grown on plastic-coated UV beads. Example 4. Phenotypic changes and inhibition of Campylobacter jejuni NCTC 11168 binding to histo-blood group antigens by Fe-QA Materials and Methods
[513] Binding of C. jejuni NCTC 11168 to BgAgs (common ABO antigens of the histo-blood group), Leb and H-II was measured after growth of the bacteria in a medium that has Fe-QA at 0.34 mM or 3 .4 mM. Binding is measured by washing the Fe-Q-containing medium prior to testing bacteria by ELISA as described below. Binding was measured after one passage and four passages (4 generations) with Fe-QA included in the medium, and compared to a control without Fe-QA.
[514]The binding of C. jejuni 11168 to BgAgs (common ABO antigens of the histo-blood group) that are expressed, for example, on the surfaces of erythrocytes, and the inhibition of this binding by Fe-QA was quantified using the ELISA method Described below.
[515] The ELISA was performed as follows: BSA-BgAg conjugates were obtained from IsoSep, Tullinge, Sweden. Coupling of BgAgs to 96-well plates (NUNC Immobilizer Amino) was performed by adding 100 µl of BSA-BgAg (5 µg/ml, unless otherwise indicated) in sodium carbonate buffer to each well. Plates were incubated at room temperature for 2 h before unbound reagent was washed away three times in PBS-T. All wells were blocked by the addition of 100 ml of 1% BSA/PBS and incubated for 2 h at room temperature. After washing in PBS-T, 100 ml of DIG-targeted bacteria (at OD600 of 0.05) was added to each well and incubated overnight at 4°C. The plates were washed three times in PBS-T before adding 100 ml of anti-digoxigenin-POD solution (Roche Diagnostics; 1 in 5000 diluted in 1% BSA/PBS) and incubated for 1 h at room temperature. The plates were again washed vigorously in PBS-T and the color developed by adding 100 µl of ABTS substrate (Roche). The plates were read with an ELISA reader (Biotek EL800) at an absorbance of 405 nm. Specific binding was determined by subtracting the binding of each strain with BSA (typically DO405 0.07-0.09) from the binding of each BSA-BgAg conjugate. Inhibition assays shown in Figure 4B were performed as above, but after removal of the blocking solution, the DIG-labeled C. jejuni was pre-incubated for 4 h with a solution of Fe-QA (0.34 mM) before be added to each well. Results
[516]Figure 4 shows binding of C. jejuni to BgAgs (common ABO antigens of the blood histo group), Leb and H-II after growth of the bacteria in a medium that has either 0.34 mM Fe-AQ or 3.4 mM. The results show a marked decrease in binding to Leb and H-II particularly in the group that was treated with the highest concentration of Fe-QA (3.4 mM). There was also a statistically significant decrease in the lowest Fe-QA concentration of 0.34 mM when compared to the control group. It is therefore evident that treating bacteria with Fe-QA for several generations results in a phenotypic change, and the bacteria lose the ability to bind BgAg permanently.
[517]Figure 4B shows the binding of C. jejuni 11168 to BgAgs (common ABO antigens of the histo-blood group) that are expressed, for example, on the surfaces of erythrocytes, and the inhibition of this binding by Fe-QA. Binding is quantified using the ELISA method described below. Bar graphs show binding for untreated C. jejuni 11168-NT (NT = untreated) to BgAgs, and significant inhibition of binding by pre-incubating the bacteria with Fe-QA before adding the bacteria to the ELISA plate. Example 5. Inhibition of Helicobacter pylori 17875 CCUG binding to human gastric tissue Materials and Methods
[518]The ability of H. pylori CCUG 17875 to bind to human gastric tissue samples in the presence of Lewis b antigen, Leb, and two concentrations of Fe-QA (1 mM and 0.2 mM) was measured. Binding was quantified by fluorescence using H. pylori that had been labeled with fluorescein, using FITC (fluorescein isothiocyanate) and human gastric tissue that was embedded in parafilm. Bacteria were suspended in blocking buffer (1% BSA in PBS) and applied to rehydrate human gastric tissue histosections. Binding was assessed microscopically, and quantified as the average number of bacteria bound to the tissue.
[519]The binding of Leb to the H. pylori strain 17875 at pH 7.4 was assessed by labeling Leb with radioactive iodine (I-125), mixing the radioactive antigen with the H. pylori bacterium, pelleting the bacterium using a centrifuge and measuring radioactivity in the pellet and supernatant. Any Leb that is bound to the bacteria is quantified by measuring the radioactivity in the pellet. The ratio of radioactivity in the pellet to the supernatant therefore corresponds to the ratio of Leb bound by the bacterium which remains unbound, expressed as bound/free. Results
[520] The ability of H. pylori to bind to human gastric tissue samples in the presence of Lewis b antigen, Leb, and two concentrations of Fe-QA (1 mM and 0.2 mM) are shown in Figure 5A. The results show that binding of H. pylori to human gastric tissue is significantly reduced in the presence of Leb (10μg/ml) and Fe-QA at concentrations of 1 mM and 0.2 mM when incubated for 1 hour at room temperature. The reduction in bacterial binding was estimated by counting the number of bacteria specifically attached to the gastric pit region under 200X magnification. Therefore, Fe-QA prevents bacterial attachment of H. pylori to the gastric epithelium.
[521]Figure 5B shows competitive inhibition of Leb binding to H. pylori by Fe-QA as the concentration of Fe-QA is increased. The graph is a graphical representation of the ratio of bound and free Leb versus Fe-QA concentration (μM). The graph shows that Fe-QA increasingly inhibits Leb binding to H. pylori as the amount of Fe-QA is increased. Example 6. Prevention of Fe-QA from biofilm formation by Pseudomonas aeruginosa and Uropathogenic E. coli (UPEC). Materials and methods
[522]APseudomonas aeruginosa PAO-1, and a clinically isolated uropathogenic Escherichia coli UPEC-536 were routinely grown on LB agar plates (Luria-Bertani, Oxoid, UK) at 37°C or in broth at 37°C with shaking at 200 rpm. UV-sterilized glass slides were incubated in 15 mL of RPMI-1640 defined medium (Sigma, UK) or 15 mL of RPMI-1640 with Fe-QA inoculated with diluted bacteria (OD600 = 0.01) from cultures during overnight at 37°C with stirring at 60 rpm for 72 hours. Slides were removed from the bacterial culture and washed with 15 ml of phosphate-buffered saline at room temperature for 5 minutes three times and then washed with distilled water H2O. After washing, slides were stained with 20 µM of SYTO17 dye (Invitrogen, UK) at room temperature for 30 minutes. After removal of excess staining dye and air drying, samples were examined using a Carl Zeiss LSM 700 Laser Scanning Microscope with ZEN 2009 imaging software (Carl Zeiss, Germany). The bacterial coverage rate on the surface was analyzed using Open Source Image J 1.44 software (National Institute of Health, USA). Results
[523]Figure 6A shows that Fe-QA ("X") at 100 μM inhibits biofilm formation by Pseudomonas aeruginosa. In the absence of Fe-QA, a higher coverage rate was measured for Pseudomonas aeruginosa than in the presence of a 100 μM concentration of Fe-QA.
[524]Figure 6B shows that Fe-QA inhibits biofilm formation by Uropathogenic E. coli (UPEC). In the absence of Fe-Q ("0 μM), a higher coverage rate is measured for UPEC than in the presence of 0.1 μM, 1 μM, 10 μM and 100 μM concentrations of Fe-QA. Example 7. Planktonic growth of bacteria in the presence of Fe-QA Materials and Methods
[525]The growth rate of Uropathogenic E. coli UPEC-536 in RPMI-1640 medium over a 24-hour period was compared to the growth rate of UPEC in RPMI-1640 medium, but in the presence of 100 μM Fe- QA The growth rate of Pseudomonas aeruginosa in RPMI-1640 medium was also compared with the growth rate of Pseudomonas aeruginosa in RPMI-1640 medium, but in the presence of 100 μM Fe-QA. Results
[526] Figure 7A is a graph showing the growth rate of UPEC in RPMI-1640 medium over a 24 hour period. The growth rate is compared to the growth rate of UPEC in RPMI-1640 medium, but in the presence of 100 μM Fe-QA. (Optical absorbance of RPMI-1640 medium is also shown for reference). The graph demonstrates that Fe-QA does not inhibit the growth of UPEC. However, as shown in Example 6 and Figure 6B, Fe-QA inhibits biofilm formation. Therefore, the inhibition of biofilm formation is not due to the inhibition of bacterial growth.
[527] Figure 7B is a graph showing the growth rate of Pseudomonas aeruginosa in RPMI-1640 medium. The growth rate is compared to the growth rate of Pseudomonas aeruginosa in RPMI-1640 medium, but in the presence of 100 μM Fe-QA. (Optical absorbance of RPMI-1640 medium is also shown for reference). The graph demonstrates that Fe-QA does not inhibit the growth of Pseudomonas aeruginosa as was also found for UPEC (above). However, as shown in Example 6 and Figure 6A, Fe-QA inhibits Pseudomonas aeruginosa biofilm formation. Therefore, the inhibition of biofilm formation is not due to the inhibition of bacterial growth. Example 8. Planktonic growth versus biofilm growth of C. jejuni mutant MOMP T268G Materials and Methods
[528] A MOMP-T Campylobacter jejuni NCTC 11168 strain was prepared by mutating T268 from MOMP. MOMP's T268 was replaced by glycine. The planktonic growth of the MOMP-T strain of Campylobacter jejuni and its ability to form a biofilm compared to the wild-type strain were determined. Bacteria (wild type and MOMP-T strain) were cultured on CCDA selective charcoal agar and then transferred to Mueller-Hinton broth (MHB). Growth suspensions were prepared at 0.02 OD/ml and then allowed to grow at 37°C under atmospheric conditions (5% CO 2 , 6% O 2 ) for 48h in MHB with plastic coated UV beads (Lascells). After 48 hours, 10 µl of suspension was serially diluted 10-fold at 10-3, 10-4, 10-5, 10-6, 10-7 and 10-8. For each dilution, 10 μl were placed on CCDA selective agar plates and colonies were counted after 48 h. The beads were also removed and washed in PBS before addition to 1 ml of PBS. After vortexing, 10 µl of the cell suspension was serially diluted as above and cell counts were performed to quantify biofilm formation. Results
[529]Figure 8A shows that the MOMP T268 mutation has no impact on the growth of Campylobacter jejuni over wild-type, although MOMP T268 is known to be O-glycosylated. However, Figure 8B shows that the MOMP T268 mutation has an impact on biofilm formation. Experience demonstrates that O-glycosylation of MOMP to T268 in Campylobacter is vital for biofilm formation, and the need for the MOMP-T268 glycan for aggregation and biofilm formation. Example 9. Fe-QA treatment causes antibiotic resistant strain of E. coli to lose antibiotic resistance Materials and Methods
[530] A kanamycin resistance gene was introduced into the genome of EPEC Enteropathogenic E. coli, (E2348/69), genotype; wild-type EPEC-O-17:H6 allows the kanamycin-resistant strain of EPEC to grow in the presence of a dressing that has been dipped in kanamycin (at a concentration of 50 μg/mL). The kanamycin-resistant strain of EPEC was then soaked in a dressing in a solution of Fe-QA. Results
[531] Soaking a patch in a Fe-QA solution and then placing the patch on a growth plate with the kanamycin-resistant strain also had no significant effect on bacterial growth. However, immersing the patch in a solution of Fe-QA and kanamycin and then placing it on a growth plate caused the bacteria to fail to grow. Experience shows that Fe-QA has no significant impact on bacterial growth. However, Fe-QA makes bacteria more sensitive to kanamycin, even though the bacteria carry a kanamycin resistant gene, and are therefore resistant to the antibiotic. Fe-QA can therefore be used in conjunction with antibiotics to kill or inhibit the growth of antibiotic resistant bacteria.
[532]Figure 9 is a graph showing the impact on the growth curve of the kanamycin-resistant EPEC strain when grown in the presence of: (i) kanamycin (top row number 2, (triangles)), (ii) Fe- QA (line number 1, from top), and (iii) kanamycin and Fe-QA (line number 3, from top (squares)). The fastest growth was found when the strain was grown in a medium containing only Fe-QA. However, a difference was observed between the growth rate of the strain in the presence of kanamycin versus kanamycin and Fe-QA. The double arrow indicates the difference in rates between these two growth conditions. The strain growth rate was inhibited in the presence of kanamycin and Fe-Q compared to the strain growth rate only in the presence of kanamycin. This is further evidence that Fe-QA can be used in conjunction with antibiotics to kill or inhibit the growth of antibiotic-resistant bacteria. Example 10. Fe-QA treatment causes antibiotic resistant strain C. jejuni NCYC 11168 to lose antibiotic resistance Materials and Methods
[533] A patch immersed in Fe-QA solution was placed on a growth plate with a wild-type Campylobacter jejuni NCTC 11168 (which does not carry a kanamycin resistance gene). Wild-type C. jejuni grew normally, and was unaffected by FeQ. A patch was also immersed in a solution of kanamycin and Fe-QA, and placed in a wild-type C. jejuni growth plate. The growth of C. jejuni was impacted showing that wild-type C. jejuni is sensitive to the presence of kanamycin.
[534]The patch experiments were repeated with a Campylobacter jejuni NCTC 11168 strain carrying a kanamycin resistance gene, where the patches were immersed in (i) kanamycin only, (ii) Fe-QA only, and (iii) kanamycin and Fe-QA. The growth of the C. jejuni strain carrying the kanamycin resistance gene was not affected by dressings immersed in (i) kanamycin and (ii) Fe-QA, however, growth was affected by the dressing that was immersed in both kanamycin and (ii) Fe-QA. as in Fe-QA. The results further demonstrate that Fe-QA can be used in combination with antibiotics to inhibit or kill antibiotic-resistant bacteria. Example 11. Fe-Tyr treatment causes antibiotic resistant E. coli (EPEC) strain E2348/69 to lose antibiotic resistance Materials and Methods
[535] The impact of the growth curve of Enteropathogenic E. coli (EPEC) strain E2348/69 (EPEC wild-type O17:H6 genotype) when cultured in the presence of (i) gentamicin (1.25 μM), (ii ) Fe-Tyr (100 μM), (iii) gentamicin (1.25 μM) and Fe-Tyr (1.25 μM), and (iv) a control without gentamicin or Fe-Tyr present was determined. Results
[536]Figure 10A shows the impact on the growth curve of the antibiotic resistant Enteropathogenic strain E. coli (EPEC) strain E2348/69 (EPEC wild-type O17:H6 genotype) when grown in the presence of (i) gentamicin (1 .25 μM) (gray circles), (ii) Fe-Tyr (100 μM) (inverted gray triangle), (iii) gentamicin (1.25 μM) and Fe-Tyr (1.25 μM) (vertical white triangle) , and (iv) a control without gentamicin or Fe-Tyr present (black circles). The fastest growth (see Figure 10B) was found when the strain was grown in medium containing only Fe-Tyr, which was comparable to the control (no gentamicin or Fe-Tyr present). However, a difference was observed between the growth rate of the strain in the presence of the antibiotic gentamicin compared to gentamicin and Fe-Tyr. Thus, it was demonstrated that the growth rate of the strain was inhibited in the presence of gentamicin and Fe-Tyr in relation to the growth rate of the strain only in the presence of gentamicin. This is further evidence that Fe-Tyr can be used in conjunction with antibiotics to kill or inhibit the growth of antibiotic-resistant bacteria. Example 12. Treatment with FeQ (also known as “Fe-QA”) causes antibiotic-resistant Enteropathogenic E. coli (EPEC) strain E2348/69 to lose antibiotic resistance Materials and Methods
[537] The impact on the growth curve of antibiotic-resistant Enteropathogenic E. coli (EPEC) strain E2348/69 (EPEC wild-type O17:H6 genotype) when grown in the presence of a fixed concentration of gentamicin (1, 25 μM) and an increasing concentration of FeQ versus the strain grown in the presence of FeQ alone or gentamicin alone was determined. Results
[538]Figures 11A-C are graphs showing the impact on the growth curve of Enteropathogenic E. coli (EPEC) strain E2348/69 (EPEC wild-type O17:H6 genotype) when grown in the presence of a fixed concentration of gentamicin (1.25 μM) and an increasing concentration of FeQ (Fig. 11A:10μM, Fig. 11B:34 μM and Fig. 11C: 68 μM) versus the strain grown in the presence of FeQ alone or gentamicin alone. As illustrated in Figure 11D, increasing the concentration of FeQ from 10 μM to 100 μM did not affect the growth rate of the strain. However, Figure 11E clearly shows that a difference was observed between the growth rate of the strain in the presence of the antibiotic gentamicin versus the combination of gentamicin and FeQ. Thus, it was demonstrated that the growth rate of the strain was inhibited in the presence of gentamicin and FeQ in relation to the growth rate of the strain only in the presence of gentamicin. This is further evidence that Fe-Q can be used in conjunction with antibiotics to kill or inhibit the growth of antibiotic-resistant bacteria. Example 13. FeQ prevents attachment of bacteria to surfaces Materials and Methods
[539] Enteropathogenic E. coli (EPEC) E2348/69 were grown in wells for 48 hours at 37°C in the presence of FeQ (100 μM), and in the absence of FeQ (as a control). After 48 hours, the wells were washed to remove suspended cells. Crystal violet was then added to each well. The wells were then washed to remove excess dye. An acetone/ethanol mixture was then added to the wells to resuspend the cells that adhere to the plastic surface of the wells, and dissolve any dye present. The presence of dye in each well was then quantified by measuring O.D. at 570 nm. Results
[540] In the absence of FeQ, EPEC binds to the plastic surface and forms a biofilm that is readily detected by coloring with crystal violet. However, in the presence of FeQ, EPEC is unable to attach to the plastic surface and form a biofilm, and is not detected by the addition of crystal violet. Figure 12 quantitatively shows the difference in binding of EPEC cells to the surface of plastic well in the absence and presence of FeQ by measuring the optical absorbance of crystal violet that was absorbed by EPEC cells bound to the surface. At a FeQ concentration of 100 μM there is little or no binding of bacterial cells to the surface and no biofilm formation. Example 14. Treatment with FeQ (also known as "Fe-QA") causes antibiotic resistant strain of Pseudomonas aeruginosa PAO-1 to lose antibiotic resistance Materials and Methods
[541] The impact on the growth curve of an antibiotic-resistant clinical isolate of Pseudomonas aeruginosa (Clinical PAO-1) grown in the presence of kanamycin and FeQ versus the clinical isolate grown in the presence of (i) FeQ, (ii) kanamycin or (iii) no addition of kanamycin or FeQ (control) was evaluated. Results
[542]Figure 13 is a graph showing the impact on the growth curve of an antibiotic-resistant clinical isolate of Pseudomonas (Clinical PAO-1) cultured in the presence of kanamycin and FeQ versus the clinical isolate cultured in the presence of (i ) FeQ, (ii) kanamycin or (iii) without addition of kanamycin or FeQ (control). The graph shows the large reduction in the growth rate of the clinical isolate of Pseudomonas PAO when kanamycin and FeQ are added to growth media compared to either kanamycin or FeQ isolates. The example demonstrates that FeQ causes the clinical isolate of Pseudomonas to lose its antibiotic sensitivity. Example 15. Impact of FeQ and Campylobacter jejuni MOMP protein glycosylation site mutation on protein expression Materials and Methods
[543] To determine the role of FeQ and glycosylation in protein expression by Campylobacter, a quantitative proteomics approach was performed to determine protein content in FeQ treatment of wild-type Campylobacter jejuni NCTC 11168 (WT) and a mutant strain. (MOMPT268G), also referred to as MOMP-T, where the O-glycosylation site of MOMP had been interrupted by an amino acid substitution that prevented the glycosylation of MOMP. The expression of (i) Campylobacter jejuni 11168 wild-type (WT), (ii) Campylobacter jejuni 11168-T MOMP (MOMP-T), (iii) Campylobacter jejuni 11168 wild-type (WT) in the presence of FeQ, and (iv) Campylobacter jejuni 11168 MOMP-T in the presence of FeQ was determined using isobaric tags of relative and absolute quantification (iTRAQ). Bacterial proteins were identified using LC-MS/MS and iTRAQ, and roles assigned according to the Genetic Ontology classification system. A total of 274,533 mass spectra from all samples led to the identification of 626 peptides (ie 420 original peptides that were assigned to 160 Campylobacter proteins). Results
[544]Table 1 shows the results of iTRAQ analysis comparing protein expression ratios for (a) WT/MOMP-T, (b) WT + FeQ/WT, (c) MOMP-T + FeQ/MOMP- T and (d) WT + FeQ/MOMP-T + FeQ, where (a) is the ratio of wild-type Campylobacter jejuni 11168 to mutant Campylobacter jejuni 11168 T268G, (b) is the ratio of wild-type Campylobacter jejuni 11168 treated with FeQ for Campylobacter jejuni 11168 wild type, (c) is the ratio of Campylobacter jejuni 11168 T268G mutant treated with FeQ for Campylobacter jejuni 11168 T268G mutant, and (d) is the ratio of Campylobacter jejuni 11168 wild type treated with FeQ for Campylobacter jejuni 11168 T268G mutant treated with FeQ. Table 1. iTRAQ Analysis














































[545] The results in Table 1 show significant changes in protein expression for 34 proteins when wild-type Campylobacter jejuni NCTC11168 or mutant Campylobacter jejuni 11168 T268G are treated with FeQ (see ratios of WT+FeQ/WT and MOMP-T+FeQ /MOMP-T). The impact of the Campylobacter jejuni 268 MOMP protein mutation to glycine is also demonstrated by the ratio of wild-type Campylobacter jejuni 11168 to Campylobacter jejuni 11168 T268G mutant (WT/MOMP-T in Table 1), and showed upregulation with a greater than 1.5 for nine proteins (putative peripartitic cytochrome C, aspartate ammonase-lyase, multifunctional aminopeptidase, Succinyl-CoA ligase, DNA-binding response regulator, flagellin subunit protein FlaB, Aspartate-ammonia-lyase, protein from Fibronectin type III domain and Glucosamine-fructose-6-phosphate aminotransferase) while ten proteins showed downregulation with a ratio of less than 0.6 (nickel-dependent hydrogenase, putative amino acid transporter, Chemotaxis protein (Fragment), Putative ATP/GTP, Lipoprotein, Energy Taxi Response Protein CetB Bipartate, Outer Membrane Major Protein, Membrane Protein, Flagellin A, P membrane protein and L-lactate permease). These results at least demonstrate that glycosylation of wild-type Campylobacter jejuni 11168 affects the expression of MOMP and Flagellin A/B, which are involved in bacterial motility, adhesion to BgAgs, aggregation and biofilm formation.
[546] Comparison of the WT + FeQ / WT ratios in Table 1 shows that five proteins (Putative GMC Oxidorectuctase, Chemotaxis Protein (Fragment), Putative Amino Acid Transporter, Energy Taxi Response Protein CetB Bipartate and Ferritin) were all upregulated above 20% and ten proteins ((Format dehydrogenase, Periplasmic nitrate reductase, Putative periplasmic cytochrome, Succinyl-CoA ligase, Aspartate ammonium lyase, multifunctional aminopeptidase A, ThiC protein of thiamine biosynthesis, DNA binding response regulator , Aspartate ammonium lyase (different subunit) and Flagellin B, FlaB) were downregulated by more than 20%.Remarkably, the results demonstrate that the glycosylated MOMP of the wild-type strain was downregulated by about 20%, and Flagellin B (major subunit of flagellum) was downregulated by about 45% when wild-type Campylobacter jejuni 11168 was treated with FeQ. These results demonstrate at least that FeQ has a impact on the expression of two essential proteins involved in colonization, adhesion and motility. Example 16. Glycosylated Campylobacter dominates broiler colonization in a mixed glycosylated and non-glycosylated population of Campylobacter Materials and Methods
[547] The chickens were colonized by a mixed strain (105 cfu, 50/50) of wild-type Campylobacter jejuni 11168 (O-glycosylated) and the Campylobacter mutant MOMPT268G (non-glycosylated). The chickens were orally challenged and after 7 days post-infection cecal samples were analyzed. Results
[548] Figure 16 is a graph showing colonization of chickens by a mixed strain (105 cfu, 50/50) of wild-type Campylobacter jejuni 11168 (O-glycosylated) and Campylobacter mutant MOMP T268G (non-glycosylated) . The chickens were orally challenged and after 7 days post-infection cecal samples were analyzed. The results show that the wild-type O-glycosylated strain dominates and is capable of establishing an infection. In contrast, the non-glycosylated strain (MOMPT268G) was unable to colonize the GI tract of the chicken, and no mutant strain could be detected. Example 17. FeQ prevents biofilm formation on human teeth Materials and Methods
[549] Molar teeth were extracted from human patients and swabs were taken from each patient's mouth to obtain samples of the bacterial flora of each patient present in the mouth. The oral swabs were cultured in the laboratory using LB medium, with the aim of developing bacterial populations normally present in the mouth of each patient. The extracted teeth were washed extensively and brushed using PBS buffer and ethanol. The tooth from each patient was then placed in the bacterial culture prepared from a sample of that patient's bacterial flora and cultured aerobically for 24 hours in LB medium. The teeth were then stained with CEPLAC™ (Manx Healthcare Ltd, Warwick, UK) and washed three times with PBS-Tween (50 mL) to determine if biofilm had formed on the teeth. Results
[550] All red-stained teeth indicate the presence of biofilm on the teeth after only 24 hours. The same teeth were then cleaned using PBS buffer and ethanol, and the procedure was repeated except for 48 hours of culture in the presence of FeQ (340 μM). After 48 hours, no teeth were stained red, demonstrating that biofilms could not be established on teeth in the presence of FeQ. Example 18. Efficacy of FeQ and FeTyr to reduce Campylobacter transport in chickens and promote growth in chickens Materials and Methods
[551] A study was conducted to assess growth promotion and reduction of Campylobacter transport using FeQ and FeTyr in Ross 308 broilers with 7 treatment groups. Each treatment group comprised four replicates of 10 birds per house (40 birds/treatment group and 4 chicken houses of 10 birds/treatment group), and there were 2 control groups and 5 test groups. All test groups and one of the control groups were exposed on day 20 of the assay to dirty sand, which tested positive for Campylobacter. This method was used to provide a more natural method of challenging Campylobacter for birds. Thus, there was a positive control in which one treatment group was challenged with Campylobacter and a negative control group in which the birds were not challenged, and five treatment groups which were all challenged with Campylobacter. The total number of birds used in the 7 treatment groups was 280. Details of the treatments are given in Table 2. Treatment group 1 was a negative control where birds were only fed commercial feed and not challenged with dirty sand containing Campylobacter. Treatment group 2 was the positive control in which birds received the commercial feed and were challenged with dirty sand containing Campylobacter on day 20. Treatment group 3 received 0.22 g/L of FeQ in their drinking water and 0. 22 g/kg of FeQ in their feed throughout the trial and challenged with dirty sand containing Campylobacter on day 20. Treatment group 5 received 0.22 g/L of FeQ in their drinking water throughout the trial and was challenged with dirty sand containing Campylobacter on day 20. Treatment group 6 received 0.22 g/kg FeQ in their feed throughout the trial and was challenged with dirty sand containing Campylobacter on day 20. Treatment group 7 received 0.022 g /L of FeQ in their drinking water throughout the trial and was challenged with dirty sand containing Campylobacter on day 20. Treatment group 8 received 0.02 g/L of FeTyr in their drinking water throughout the trial and was challenged with dirty sand containing Campylobacter on day 20. FeTyr was pre-dis dissolved in DMSO and diluted to give a 0.02 g/L solution of FeTyr in water. (An additional 4 treatment group was terminated due to solubility issues.) Table 2: Treatment details

[552]Birds were fed a commercial three-phase feeding program using starter, grower and finisher starters with the formulations shown in Table 3. All diets had coccidiostat (0.0625% MAXIBAN® in starter phase diets). initiation and finishing and MONTEBAN® at 0.06% in the finishing phase). Xylanase (RONOZYME® WX at 200 g per ton) and phytase (RONOZYME® P at 150 grams per ton) were added to all diets. Table 3: Basal food formulation for starter, growth and finisher diets


[553] Feeding schedule is shown in Table 4. Birds were reared in closed houses on day 42 and fed starter, grower and finisher feed on day 0 to 11, 11 to 24 and 24 to 42 days , respectively. All birds were individually weighed and feed weights recorded at days 0, 11, 21, 24 and 42 days. Table 4. Feeding program

[554] Prior to challenging the chickens with dirty sand containing Campylobacter on day 20, each coop was tested for Campylobacter using cloacal swabs. All chicken houses tested negative for Campylobacter prior to challenge. On day 20, the sand, which was naturally contaminated with Campylobacter, was tested to confirm the presence of Campylobacter and then added (approximately 2 kg/hen house) to the litter in all chicken houses except chicken houses for treatment group 1 ( negative control). On day 28, the chicken coop sand was sampled to confirm the presence or absence of Campylobacter. On day 41 and 42, caecal samples were taken from 3 birds per coop (12 birds per treatment group) and tested for Campylobacter enumeration. On day 42, digesta, fecal samples and cecal content were taken from all birds, and pooled by chicken coop. Two birds per coop were also taken from treatment groups 1 to 3, euthanized and blood samples taken. Samples were analyzed for blood chemistry, including analysis for alkaline phosphatase, aspartate amino transferase, alanine amino transferase, gamma-glutamyl transferase, lactate dehydrogenase, total protein, albumin, globulin, amylase, and glucose.
[555]In order to minimize the risk of cross-contamination, industry standard biosecurity measures were used, including: disinfection of boots, exchange of shorts and gloves between chicken houses/treatments, entry into Campylobacter negative chicken houses before entering positive chicken houses for Campylobacter pens and leave adjacent chicken coops empty. Daily health, slaughter and mortality were recorded. All bird weights were recorded at 0, 11, 21, 24, 33 and 42 days. Weight gains, feed intake and feed conversion index (FCR) were obtained for each feeding period.
[556] The effect of the treatment groups compared to the negative control group (treatment group 1) and the positive control group (treatment group 2) is shown in Tables 5-12 for the periods 0-11 days , 11-20 days, 20-25 days, 11-25 days, 25-42 days, 20-42 days, 0-20 days and 0-42 days. Results
[557] Figure 17 shows the mean body weight at day 42 for all treatment groups and a comparison with a commercial control designated "Target". The figure shows that treatment group 1 (the negative control labeled "CNC") achieved a mean body weight (ABW) of 3,437 kg on day 42 (which was higher than the commercial target of 2,979 kg). The positive control (marked "CC"), which was challenged with dirty sand containing Campylobacter at day 20, in contrast only achieved an ABW of 3.186 kg at day 42, which was significantly lower than the negative control (treatment group 1). ). This result demonstrates that the challenge with Campylobacter contaminated dirty sand resulted in a reduction in chicken growth by an average of 251 grams. However, when chickens were challenged with dirty sand containing Campylobacter but treated with FeQ or FeTyr in treatment groups 3, 5, 6, 7 and 8, all treatment groups had better results than the positive control demonstrating that the treatment with FeQ and FeTyr had a positive effect on growth. In fact, FeQ in the feed at 0.22 g/kg (treatment group 6) produced chicken with an ABW of 3.464 kg, which was superior to the negative control ABW of 3.437 kg, despite treatment group 6 being challenged with dirty sand containing Campylobacter.
[558]Figure 18 shows the mortality-adjusted feed conversion ratio (MFCR) at day 42 for all treatment groups and a comparison with a commercial control designated "Target". (A lower MFCR number is a better outcome.) The figure shows that treatment group 1 (the "CNC" milestone negative control) had an MFCR of 1.563, which was lower than the commercial target of 1.703. The positive control, labeled "CC", which was challenged with dirty sand containing Campylobacter on day 20 had a significantly higher MFCR of 1.679 than the negative control. Thus, challenge with Campylobacter-infected dirty sand resulted in a higher MFCR. However, when chickens were challenged with Campylobacter infected dirty sand but treated with FeQ or FeTyr in treatment groups 3, 5, 6, 7 and 8, all treatment groups had better results than the positive control demonstrating that the treatment with FeQ and FeTyr had a positive effect on MFCR (ie, decreasing numerical MFCR). Results show that treatment groups 3, 5, 6, 7 and 8 had MFCR values of 1.595, 1.560, 1.563, 1.612 and 1.577, respectively. Furthermore, treatment groups 5 and 6 performed well as the negative control even when challenged with dirty sand containing Campylobacter.
[559]Figure 19 shows the number of Campylobacter colony forming units per gram (cfu/g) of bird droppings at day 42 for treatment groups 1-3 and 6-8. (A lower number is a better result.) The results show that treatment groups 3 and 6-8 performed better than the positive control (treatment group 2) demonstrating that FeQ and FeTyr had a positive effect in reducing Campylobacter infection of poultry. Notably, chickens treated with FeTyr, FeQ in feed, and FeQ in feed and water had Campylobacter colony forming units per gram of drop that were similar or lower than the negative control group (treatment group 1). The detection of low levels of Campylobacter in the negative controls demonstrates how highly contagious the bacterium is, and is likely an indication that a small number of birds in the negative control group were infected despite not being experimentally challenged with dirty sand. The results in Fig. 19 for the low concentration of FeQ in water (0.022 g/L, treatment group 7) appear to show less effect than the other treatment groups, although this difference was considered more likely due to experimental error, for example, after cross-contamination of samples. As discussed below, the results obtained from an additional experiment as indicated in Fig. 20 confirm that treatment group 7 indeed also provided the highly beneficial effect.
[560]Figure 20 shows the average number of Campylobacter colony forming units per gram (cfu/g) of game samples at day 42 for treatment groups 1-3 and 5-8. Results show that treatment groups (3 and 5-8) performed better than the positive control (treatment group 2) demonstrating that FeQ and FeTyr had a positive effect in reducing Campylobacter infection of poultry.
[561] The effect of treatments on overall livability and the European production factor and efficiency (EPEF) is shown in Table 5. (EPEF = [(Live x Live weight in kg at the end of the trial/Age in Days x commercial FCR ) x 100].
[562] The effect of FeQ treatment on growth performance in the absence of Campylobacter challenge during the initiation phase (0-11 day) and in the 0-20 day period is illustrated in Table 14. Since controls negative and positive (treatment groups 1 and 2) are identical before the dirty sand challenge at day 20, these groups can be pooled for comparison with treatment groups 3, 5, 6 and 7 to see if FeQ had an effect growth in the absence of a Campylobacter-contaminated dirty sand challenge during the first 20 days of growth. The results demonstrate that FeQ promotes chicken growth even in the absence of a Campylobacter contaminated dirty sand challenge. On day 20, the mean body weight (ABW) for the control groups (treatment groups 1 and 2) was 0.927 kg versus 0.963 kg for treatment groups 3, 5, 6 and 7, which received FeQ. This improvement in body weight is also reflected in a significantly better MFCR for the FeQ treated birds. Table 11 shows that the MFCR for birds treated in groups 3, 5, 6 and 7 is 1.2996 versus 1.3374 for the control groups (treatment groups 1 and 2). Notably, the P value is less than 0.05.
[563] The same positive effect of FeTyr treatment on growth performance in the absence of Campylobacter challenge is also evident in Table 11. The AWG during the first 20 days of production of FeTyr treated chicken (treatment group 8) is of 0.895 kg compared with 0.884 and 0.889 kg for treatment groups 1 and 2 (negative and positive control). Furthermore, the MFCR during the first 20 days of production for the FeTyr treated broiler (treatment group 8) is 1.311 versus 1.32 and 1.355 for treatment groups 1 and 2, respectively. (A lower MFCR value is an improvement.)
[564] The results of this study demonstrate that FeQ and FeTyr promote growth and decrease the mortality-adjusted feed conversion ratio (MFCR) in the absence or presence of Campylobacter contaminated dirty sand. Table 5. Effect of treatments on growth performance during the initiation phase (day 0-11)

ab within a column reflects differences between treatments when P <0.05; SED = Standard errors of difference of means; ABW = mean body weight (kg); AFD = average food consumption (kg); AWG = average weight gain (kg); MFCR = Mortality-adjusted feed conversion index; FCR = Food Conversion Index - Commercial. Table 6. Effect of treatments on growth performance during the growth phase (day 11-20)

SED = Standard errors of difference of means; ABW = mean body weight (kg); AFD = average food consumption (kg); AWG = average weight gain (kg); MFCR = mortality-adjusted HRR; FCR = commercial FCR. Table 7. Effect of treatments on growth performance during the 20-25 day period.
SED = Standard errors of difference of means; ABW = mean body weight (kg); AFD = average food consumption (kg); AWG = average weight gain (kg); MFCR = mortality-adjusted HRR; FCR = commercial FCR. Table 8. Effect of treatments on overall growth performance during the growth phase (day 1125)
SED = Standard errors of difference of means; ABW = mean body weight (kg); AFD = average food consumption (kg); AWG = average weight gain (kg); MFCR = mortality-adjusted HRR. Table 9. Effect of treatments on overall growth performance during the finishing phase (day 25-42)
ab within a column reflects differences between treatments when P <0.05; SED = Standard errors of difference of means; ABW = mean body weight (kg); AFD = average food consumption (kg); AWG = average weight gain (kg); MFCR = mortality-adjusted HRR; FCR = commercial FCR. Table 10. Effect of treatments on growth performance during the 20-42 day trial period (after bird challenge)
ab within a column reflects differences between treatments when P <0.05; SED = Standard errors of difference of means; ABW = mean body weight (kg); AFD = average food consumption (kg); AWG = average weight gain (kg); MFCR = mortality adjusted. Table 11. Total effect of treatments on growth performance during the day 0-20 trial period (before birds were challenged).
SED = Standard errors of difference of means; ABW = mean body weight (kg); AFD = average food consumption (kg); AWG = average weight gain (kg); MFCR = mortality adjusted. Table 12. Total effect of treatment groups on growth performance (day 0-42)
ab within a column reflects differences between treatments when P <0.05; SED = Standard errors of difference of means; ABW = mean body weight (kg); AFD = average food consumption (kg); AWG = average weight gain (kg); MFCR = mortality adjusted. Table 13. Effect of treatments on global habitability and on the European factor of production and efficiency (EPEF)
Table 14. Effect of treatments on growth performance in the absence of Campylobacter challenge during the initiation phase (0-11 day) and period 0-20 days.
ABW = mean body weight (kg); AFD = average food consumption (kg); AWG = average weight gain (kg); MFCR = mortality-adjusted feed conversion ratio Example 19. FeDOPA treatment causes antibiotic-resistant Enteropathogenic E. coli (EPEC) strain E2348/69 to lose antibiotic resistance Materials and Methods
[565] The impact on the growth curve of the antibiotic-resistant E. coli (EPEC) strain E2348/69 (wild-type EPEC O17:H6 genotype) when grown in the presence of a fixed concentration of gentamicin (1, 25 μM) and an increasing concentration of FeDOPA versus the strain grown in the presence of FeDOPA alone or gentamicin alone was determined. Results
[566]Figures 21A-C are graphs showing the impact on the growth curve of the Enteropathogenic E. coli (EPEC) strain E2348/69 (EPEC wild-type O17:H6 genotype) when grown in the presence of a fixed concentration of gentamicin (1.25 μM) and an increasing concentration of FeDOPA (Fig. 21A:130μM, Fig. 21B:160 μM and Fig. 11C:200 μM) versus the strain grown in the presence of FeDOPA alone or gentamicin alone. The graphs show that the strain growth rate was inhibited in the presence of gentamicin and FeDOPA in relation to the strain growth rate only in the presence of gentamicin. This is further evidence that FeDOPA can be used in conjunction with antibiotics to kill or inhibit the growth of antibiotic-resistant bacteria. Example 20. FeDOPA prevents attachment of bacteria to surfaces Materials and Methods
[567] Enteropathogenic E. coli (EPEC) E2348/69 were cultured in wells for 48 hours at 37°C in the presence of FeDOPA (10-250 μM), and in the absence of FeDOPA (as a control). After 48 hours, the wells were washed to remove suspended cells. Crystal violet was then added to each well. The wells were then washed to remove excess dye. An acetone/ethanol mixture was then added to the wells to resuspend the cells that adhere to the plastic surface of the wells, and dissolve any dye present. The presence of dye in each well was then quantified by measuring O.D. at 570 nm. Results
[568] In the absence of FeDOPA, EPEC binds to the plastic surface and forms a biofilm that is readily detected by coloring with crystal violet. However, in the presence of FeDOPA, the attachment of EPEC to the plastic surface and the formation of a biofilm are inhibited. Figure 22 quantitatively shows the difference in binding of EPEC cells to the surface of plastic well in the absence and presence of FeDOPA by measuring the optical absorbance of crystal violet that was absorbed by EPEC cells bound to the surface. At a FeDOPA concentration of 68-250 μM bacterial cell binding to the surface and biofilm formation are inhibited. Example 21. Campylobacter jejuni loses motility after treatment with FeQ.
[569]Campylobacter jejuni NCTC 11168 was treated with FeQ (34 μM) and a plate containing brain-heart infusion (BHI) medium inoculated with 5 μl, 2 x10 5 colony forming units of the treated bacteria and the plate was cultivated for 43 hours. Bacteria growth and motility after FEQ treatment were compared to a positive control, where the bacteria had not been treated with FeQ, and also to a negative control, where no bacteria were applied to a plate of BHI medium. Data (not shown) showed that after 43 hours, Campylobacter jejuni treated with FeQ had a clear zone around the bacteria indicating that the bacteria were not motile. In contrast, Campylobacter jejuni that was not treated with FEQ was motile and spread around the culture plate. There was no visible growth, as expected, on plates that were not inoculated with bacteria. The experiment demonstrates that Campylobacter jejuni loses motility after treatment with FeQ and is consistent with the results obtained by the analysis of iTRAQ which demonstrate that FeQ downregulates the expression of FlaA (Campylobacter Flagella). Example 22. Disruption of a preformed biofilm with FeTyr
[570] Crystal violet assays were used as described above to demonstrate that FeTyr can disrupt a pre-formed biofilm. A mature biofilm formed by EPEC-pgA++ was treated with FeTyr for 24, 48 and 72 hours at FeTyr concentrations of 100 μM, 150 μM and 200 μM and the presence of the biofilm after these times was compared to an untreated biofilm (marked "Control ") using a crystal violet assay. The color of the control wells was more intense in color at 72 hours than those that were treated with FeTyr at 100, 150 and 200 μM for 72 hours. Figure 23 quantitatively shows the optical absorbance of crystal violet at 570 nm that was absorbed by EPEC cells that remained attached to the surface of the plastic well after a mature biofilm formed by EPEC-pgA++ was treated with FeTyr (shown as FeY in Fig. 23) at 100 μM, 150 μM and 200 μM compared to an untreated biofilm (labeled "Control") in the crystal violet assay. A significantly lower optical absorbance was found at 72 hours for the biofilm treated with FeTyr at 100, 150 and 200 μM at 72 hours. These results demonstrate that FeTyr can disrupt a preformed biofilm. Example 23. Disruption of a preformed biofilm with FeTyr and Fe-DOPA
[571] A BioFlux system was used to demonstrate that FeTyr and Fe-DOPA can be used to disrupt a mature EPEC-ΔcsrA biofilm. Our studies showed that a mature biofilm of EPEC-ΔcsrA can be formed in the presence of LB medium at 30% v/v and imaged (data not shown). The mature biofilm was treated with FeTyr at concentrations of 100, 150 and 200 μM for 20 hours and compared to a control biofilm that had just been treated with LB medium at 30% v/v. Biofilm dispersion was found to increase as the concentration of FeTyr increased from 100 to 200 μM (data not shown). The mature biofilm was treated with FeDOPA at a concentration of 100 μM for 20 hours and compared to a control biofilm that had just been treated with LB medium at 30% v/v. Fe-DOPA was found to disperse the biofilm at a concentration of 100 μM (data not shown). Example 24. Treatment of an acne patient with FeQ
[572] An 18-year-old female patient was treated continuously for 30 days by applying a ferric quinate solution (340 μM) to acne vulgaris ("acne") once daily. Within 5 days of starting treatment, his acne, which had not previously responded to antibiotic treatment, began to show signs of healing. After treatment for 30 days, the acne was completely cured. Acne did not recur even after discontinuation of treatment for more than a year. . Example 25. Effect of FEQ on biofilm formation of a medical device Materials and Methods
[573] To investigate the effect of Ferric Quinate (FeQ) on the surface integrity of contact lenses, two contact lenses were independently incubated in saline or saline with a final concentration of FeQ at 340μM at 4°C for 7 days, shaking gently. The lenses were then washed 6 times with phosphate buffered solution (PBS) + 0.05% Tween. Each lens was then washed twice with distilled water prior to environmental scanning electron microscope (ESEM) analysis.
[574] To investigate biofilm formation on contact lenses, clinically determined Pseudomonas aeruginosa strains PAO-1 were incubated with the lenses in Luria-Bertani Media (LB) or LB with a final concentration of FeQ at 340μM at 37°C for 24 hours in an unstirred incubator. The lenses were then washed 6 times with PBS + 0.05% Tween, before being stored overnight in PBS + 0.05% Tween. In preparation for the ESEM, formaldehyde was added to a final concentration of 1% and incubated for 10 minutes to inactivate the bacteria. The lenses were then washed 4 times with PBS + 0.05% Tween, and immediately before analysis the lenses were washed two more times with distilled water. Results Surface integrity
[575] Lenses were either FeQ treated at 340 μM or untreated (control) to investigate the effect, if any, of FeQ treatment on the surface integrity of the contact lens.
[576] The results show that FeQ at 340 μM has no visible significant effect (via ESEM) on the surface integrity of the contact lens. Biofilm formation
[577]ESEM images (data not shown) after incubation with bacteria showed only large-scale biofilm development of Pseudomonas aeruginosa on the surface of contact lenses. Analysis via ESEM requires a vacuum and causes areas of the biofilm to dehydrate, which is responsible for the perforated appearance of the contact lens biofilm. Dehydration gives depth perception, and shows the biofilm formed in the absence of FeQ to be substantial. Individual bacteria were visible in the biofilm, surrounded by the extracellular matrix (ECM).
[578] The impact of incubation with bacteria in the presence of FeQ 340μM was also investigated. These images (data not shown) showed that in the presence of FeQ at 340 μM, Pseudomonas aeruginosa appears as a single bacterium, or small aggregates of bacteria, with no apparent ECM formation. These results indicate that FeQ inhibits Pseudomonas aeruginosa biofilm formation.
[579] Based on these results, it can be concluded that FeQ and other compounds as described herein can be used to inhibit or prevent biofilm formation in medical devices such as, but not limited to, contact lenses. Example 26: Metabolomic Analysis Materials and Methods
[580]The strains analyzed were wild-type Campylobacter jejuni NCTC 11168, and a mutant (MOMP268T/G) where the wild-type MOMP protein Thr-268 is mutated to glycine, resulting in a strain of the bacteria in which the MOMP protein cannot be glycosylated. The mutant is further described in WO 2013/121214. The strains were cultivated for 48 hours in Mueller Hinton Broth (MHB). In samples without control, bacteria (wild type and mutant) were treated with FeQ at a concentration of 340 μM.
[581] Three replicates of one sample were taken from each culture and each replicate was analyzed three times (ie, yielding 9 readings for each sample). Thus, for example, the sample taken from fresh medium (FM) produced nine readings, labeled FM- 1_1, 1_2, 1_3, 2_1, 2_2, 2_3, 3_1, 3_2 and 3_3, respectively. It is the same for each of the other samples, which are: SMWT: spent medium from the wild-type control culture SMWTF: spent medium from the wild-type culture that was grown in the presence of FeQ SMMT: spent medium from from SMMTF mutant control culture: spent medium from the mutant culture that was grown in the presence of FeQ For the metabolite profile, LC was performed on an Accela system (Thermo Fisher Scientific, Hemel Hempstead, UK). Chromatographic separation was performed using a ZIC-pHILIC column (150 mm x 4.6 mm, 5 µm column, Merck Sequant) as previously described (Creek et al. 2011, Anal Chem 83, 8703-8710). Briefly, the column was kept at 45°C and samples were eluted with a linear gradient from 80% B to 5% B over 15 min, followed by a 2 min linear gradient from 5% B to 80 % B and 7 min re-equilibration with 80% B at a flow rate of 300 μl/min. Mobile phase A was 20 mM ammonium carbonate in water and mobile phase B was 100% acetonitrile acetonitrile. The injection volume was 10 μl and the samples were kept at 4°C. An Orbitrap Exactive (Thermo Fisher Scientific, Hemel Hempstead, UK) with a HESI 2 probe was operated in polarity switching mode, with the following settings: resolution 50 000, AGC 1 x 106, m/z range 70 - 1400, gas sheath 40, auxiliary gas 5, sweep gas 1, probe temperature 150°C and capillary temperature 275°C. For positive mode ionization: Source voltage +4.5 kV, capillary voltage +50 V, tube voltage +70 kV, skimmer voltage +20 V. For negative mode ionization: Source voltage -3.5 kV, capillary voltage -50V, tube voltage -70V, skimmer voltage -20V. Mass calibration was performed for each polarity immediately before each batch of analysis. The calibration mass range has been extended to cover small metabolites by including low mass contaminants with Thermo calibration mix standard masses (below m/z 1400), C2H6NO2 for positive ion electrospray ionization mode ( PIESI) (m/z 76.0393) and C3H5O3 for negative ion electrospray ionization (NIESI) (m/z 89.0244). Data processing and analysis
[582] Raw LC-MS data were processed with XCMS for untargeted selection peaks (Tautenhahn et al. 2008, BMC Bioinformatics 9, 504) and mzMatch. R for peak matching and annotation of related peaks (Scheltema et al. 2011, Analytical Chemistry 83, 2786-2793). Putative identification of metabolites was performed by IDIOM using standard parameters (Creek et al. 2012, Analytical Chemistry 84, 8442-8447). Metabolite identification was performed by matching accurate masses and retention times to authentic standards (Level 1 metabolite identification according to the Metabolomics Standards Initiative (Sumner et al. 2014, Metabolomics 10, 1047-1049; Sumner et al. 2007). , Metabolomics
[583] 211-221). However, when standards were not available, predicted retention times were used, so these identifications should be considered putative (level 2 identification). Results and conclusions:
[583]A Fig. Fig. 24A shows the analysis data in a positive way, as a graph of OPLS-DA scores. This shows a clear separation between fresh media (FM) and other used media (SMWT, SMWTF, SMMT, SMMTF) which demonstrates that many metabolites were excreted and consumed during cell culture.
[584]Fig. 24B also shows the analysis data in a positive way. The fresh medium (FM) results have been removed from the graph because they are so different from the other samples that any differences between the different spent medium samples could be hidden. This graph shows a clear separation between wild-type (SMWT) and wild-type + FeQ (SMMTF), but SMMT and SMMTF grouped more closely together. This indicates that FeQ does not cause a large detectable change between mutant and mutant + FeQ. In general, it is clear that there are fewer metabolic changes caused by FeQ in the mutant than in the wild type.
[585] Fig. 24C contrasts with Fig. 24A in that it shows the analysis data in a negative way, although essentially the same pattern and conclusions apply as in Fig. 24A.
[586] Fig. 24D contrasts with Fig. 24B in that it shows the analysis data in a negative way. The negative mode data in Fig. 24D shows slightly different trends from the positive mode data and demonstrates a clear separation between mutant (SMMT) and mutant + FeQ (SMMTF) samples, as well as between SMWT and SMWTF samples.
[587] These data demonstrate how fundamentally bacterial metabolism is altered by treatment with FeQ. This is consistent with the phenotypic changes observed in FeQ-treated bacteria (as confirmed by the iTRAQ results as discussed in Example 15), and provides insight into the mechanism underlying the capacity of FeQ and its related compounds as discussed in Section III.A. of this application to treat bacteria and cause an inhibition in their ability to form biofilms, colonize chickens and other animals, and even make bacteria less resistant to antibiotics. Example 27: Preparation protocol for K[Fe(C7H11O6)3](OH)3H2O(FeQ)
[588] FeCl3 • 6H2O (50 g, 184 mmol, Alfa Aesar, 97%) was placed in a flask and dissolved in 300 mL of H2O (J.T. baker, HPLC class). To this solution, D-(-)-quinic acid (110 g, 572 mmol, Buchlr Gmbh, 96%) was slowly added with continuous stirring. The pH of the solution was adjusted to ~3 by adding 10M KOH (Alpha Aesar, 85%) (~80 mL was required).
[589] The dark yellow solution was darkened to brownish by addition of KOH. The dark solution was stirred at room temperature for 1 hour. After stirring at room temperature for 1 h, ethanol (EMD, 94%) (2.5 L) was slowly added to the solution with stirring.
[590] After approximately 1/4 of the total ethanol was added, the solution visibly lit up and a fine solid began to precipitate from the solution. After the remaining ethanol is added, the solution is allowed to stand overnight at room temperature.
[591] The solids are collected by vacuum filtration on a fritted funnel and allowed to dry on the funnel while the vacuum is continued for 2-3 hours. The bright yellow solid is spread out in a thin layer on a drying dish and air dried for 3 days followed by vacuum drying for 48 h to give 155 g of the final product. Example 28: Synthesis of Fe (Tyr)3
[592] L-tyrosine (5.43 g, 30 mmol, Chem Impex, 99.5%) and LiOH 2 H 2 O (1.26 g, 30 mmol, EMD, 94%) were dissolved in water (250 ml, JT Baker, HPLC class)) and the solution heated at 70°C for 20 min. The FeCl 3 salt (1.62 g, 10 mmol, Alpha Aesar, 98%) was dissolved in a minimal amount of water (3-5 ml) and added to the tyrosine/LiOH solution.
[593] Precipitation (brown solid) was almost instantaneous, but stirring with heating continued for 15 min. The product was allowed to cool to room temperature and collected by filtration. The product was air dried and then further dried in a lyophilizer. The isolated yield was 5.85 g. Example 29: Synthesis of Fe (DOPA)3
[594] L-Dopa (11.84 g, 60 mmol, AK Scientific, 98%) and LiOH 2 H 2 O (2.52 g, 60 mmol, EMD, 94%) were dissolved in water (100 ml, JT Baker, HPLC class)) and the solution heated at 70°C for 20 min. The FeCl 3 salt (3.2 g, 20 mmol, Alpha Aesar, 98%) was dissolved in a minimal amount of water (6-10 ml) and added to the vigorous Dopa/LiOH solution.
[595] Precipitation (very dark purple) was almost instantaneous, but stirring with heating continued for 15 min.
[596] The product was allowed to cool to room temperature and was collected by filtration. The product was air dried and then further dried in a lyophilizer. The isolated yield was 6.5 g. More solid (4 g) was collected from the filtrate and dried in the same manner. The overall yield was 10.5 g. Example 30: Fe-Q and Fe-Phe potentiate the effect of antibiotics Methods
[597] To investigate the effects on antibiotic resistance of a laboratory strain of Psuedomonas aeruginosa (PAO1N) and a mixed population of clinical isolates (PAO Mixed), when incubated in Luria-Bertani (LB) medium alone, or at different concentrations (34 μM, 100 μM, 200 μM and 340 μM) of FeQ or FePhe.
[598] Each of the different media, a control bar, contained 10 μg/ml of the aminoglycoside antibiotic Amikacin.
[599]10 µl of the bacterial strains were added to each well of a 96-well microtiter plate, before 290 µl of the relevant media was added to the wells. Each different condition was repeated in sextuplicate.
[599] The plate was incubated at 37°C inside a microtiter plate reader for 17.5 hours, with the OD 600 read every 30 minutes. Results
[601] Results are shown in FIG. 25A and 25B. These values show that Fe-Q and Fe-Phe provide similar effects in reducing the tolerance of PAO1N and PAO Mixed to the aminoglycoside Amikacin.

权利要求:
Claims (16)
[0001]
1. A non-therapeutic method of increasing the growth of an animal selected from a non-human mammal or bird, the method comprising causing the animal to ingest or absorb an effective amount of one or more compounds, wherein the one or more compounds are Fe III complexes comprising ligands attached to the iron center selected from amino acids and α-hydroxy acids or a salt and/or hydrate thereof, with the proviso that the one or more compounds is not a quinic acid complex with Fe III, and wherein the increased growth of the animal includes an increase in one or more characteristics selected from the group consisting of increasing body weight or (in the case of a group of animals) average body weight (ABW ), feed intake or (in the case of a group of animals) the mean feed intake (AFD), the weight gain or (in the case of a group of animals) the mean weight gain (AWG) and/or the feed conversion ratio adjusted to dead ity (MFCR).
[0002]
2. Non-therapeutic use of one or more compounds, characterized in that the one or more compounds are Fe III complexes comprising ligands attached to the iron center selected from amino acids and α-hydroxy acids or a salt and/or hydrate thereof, with the proviso that the one or more compounds is not a complex of quinic acid with Fe III, to enhance the growth of an animal selected from a non-human mammal or bird by causing the animal to ingest or absorbs an effective amount of one or more compounds, wherein the increased growth of the animal includes an increase in one or more characteristics selected from the group consisting of increasing body weight or (in the case of a group of animals) average body weight (ABW), the feed intake or (in the case of a group of animals) the mean feed intake (AFD), the weight gain or (in the case of a group of animals) the mean weight gain (AWG) and /or the mortality-adjusted feed conversion index ade (MFCR).
[0003]
3. Method according to claim 1, or use according to claim 2, characterized in that the one or more Fe III complexes comprise ligands attached to the iron center selected from the group consisting of: (a) one or more amino acids selected from the group consisting of alanine, arginine, asparagine, aspartic acid, cysteine, glutamine, glutamic acid, glycine, histidine, isoleucine, leucine, lysine, methionine, phenylalanine, proline, serine, threonine , tryptophan, tyrosine and valine or a salt and/or hydrate thereof; and (b) one or more α-hydroxy acids selected from the group consisting of lactic acid, glycolic acid, citric acid, and mandelic acid or a salt and/or hydrate thereof.
[0004]
4. Method according to claim 1 or 3, or use according to claim 2 or 3, characterized in that the one or more compounds are presented to the animal through one or more routes selected from the group which consists of an animal feed, an animal feed supplement, and water.
[0005]
5. Method according to any one of claims 1, 3 or 4, or use according to any one of claims 2 to 4, characterized in that the animal is selected from the group consisting of: (a ) poultry, (b) poultry selected from chicken, turkey or ducks; (c) livestock; or (d) selected livestock from cattle, sheep, goats or swine.
[0006]
6. Method or use, according to claim 5, characterized in that the animal is: (a) a chicken, (b) a meat type chicken; (c) a beef type chicken; (d) an egg hen; (e) an egg hen which is a pullet or hen (f) a breeder hen.
[0007]
7. Method according to claim 1 or use according to claim 2, characterized in that the method or use additionally comprises the step of rearing the animal to allow for increased growth, wherein the animal is a non-human animal , wherein the method or use further comprises the step of harvesting a product from the non-human animal bred with increased growth, and wherein: (a) the harvested product is a by-product of the non-human animal, (b) the harvested product is a non-human animal by-product selected from milk, eggs, or wool; (c) the harvested product is the body or part of the body of the non-human animal, and the harvesting process includes the stage of slaughtering the non-human animal, (d) the harvested product is the body or part of the body of the non-human animal and the harvesting process includes the stage of slaughtering the non-human animal and preparing: (i) a non-human animal carcass or part thereof, as a product, or (ii) a non-human animal carcass or part of the even as a meat product.
[0008]
8. Method or use according to claim 7, characterized in that the animal is a chicken, for example, (a) a meat type chicken, (b) a meat type broiler, (c) a hen, (d) a pullet of eggs, or (e) a pullet.
[0009]
9. Method, or use, according to claim 8, characterized in that the animal is a broiler chicken and the method or use comprises the stage of slaughtering the animal before the age of 47, 46, 45, 44, 43, 42, 41, 40, 39, 38, 37, 36, 35, 34, 33, 32, 31, 30, 29, 28, 27, 26, 25, 24, 23, 22, 21 or 20 days.
[0010]
10. Method, or use, according to claim 9, characterized in that the broiler has reached a target body weight at the time of slaughter.
[0011]
11. Method according to any one of claims 1 or 3 to 10, or use according to any one of claims 2 to 10, characterized in that: (a) the animal to be treated is not infected with or disadvantageously colonized by bacteria or other microorganisms and/or in any case healthy; and/or (b) the method or use comprises the eventual slaughter of the non-human animal.
[0012]
12. Method according to any one of claims 1 or 3 to 11, or use according to any one of claims 2 to 11, characterized in that the method or use comprises the eventual slaughter of the animal, in which the animal is a non-human animal, and the preparation of an animal carcass, or part thereof, as a product.
[0013]
13. Method according to claim 12, or use according to claim 12, characterized in that the product is a meat product.
[0014]
14. Feed suitable for use in the method as defined in any one of claims 1 or 3 to 13, or for use as defined in any one of claims 2 to 13, characterized in that the feed comprises or is supplemented with, one or more compounds which are Fe III complexes comprising ligands attached to the iron core selected from amino acids and α-hydroxy acids or a salt and/or hydrate thereof, wherein the one or more Fe III complexes comprise ligands attached to the iron core selected from the group consisting of: (a) one or more amino acids selected from the group consisting of alanine, arginine, asparagine, aspartic acid, cysteine, glutamine, glutamic acid, histidine, isoleucine , leucine, lysine, methionine, phenylalanine, proline, serine, threonine, tryptophan, and valine; and (b) one or more α-hydroxy acids selected from the group consisting of lactic acid, glycolic acid, citric acid, and mandelic acid, or a salt and/or hydrate thereof; and wherein the animal feed comprises or is supplemented with the one or more compounds in an amount from 0.001 to 20 g of the one or more compounds per kg of feed.
[0015]
15. Animal food according to claim 14, the method according to any one of claims 1 or 3 to 13, or the use as defined in any one of claims 2 to 13, characterized in that the one or more Fe III complexes are provided in an animal feed, and: (a) the feed is a chicken feed; (b) the feed is a chicken feed selected from a starter diet, grower diet and finisher diet; (c) the feed is a chicken feed as defined by (a) or (b) and the chicken is selected from the group consisting of a meat type hen, a meat type broiler hen, an egg hen, a laying hen selected from a pullet or hen, and a breeding hen; (d) the feed is a poultry feed, or a poultry feed for turkeys or ducks; (e) the feed is livestock feed; (f) the feed is a feed for cattle, sheep, goats or swine; (g) the food is for domestic animals; (h) the food is cat or dog food; (i) the food is food for horses.
[0016]
16. Supply of drinking water for animals suitable for use in the method as defined in any one of claims 1 or 3 to 13, or use as defined in any one of claims 2 to 13, characterized in that the drinking water comprises or is supplemented with one or more compounds which are Fe III complexes comprising ligands attached to the iron core selected from amino acids and α-hydroxy acids or a salt and/or hydrate thereof, wherein the one or more Fe III complexes comprise ligands linked to the iron center selected from the group consisting of: (a) one or more amino acids selected from the group consisting of alanine, arginine, asparagine, aspartic acid, cysteine, glutamine, glutamic acid, histidine, isoleucine, leucine , lysine, methionine, phenylalanine, proline, serine, threonine, tryptophan and valine; and (b) one or more α-hydroxy acids selected from the group consisting of lactic acid, glycolic acid, citric acid, and mandelic acid, or a salt and/or hydrate thereof; and wherein the drinking water comprises or is supplemented with the one or more compounds in an amount from 0.001 to 20 g of the one or more compounds per L of water.

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引用文献:

---

公开号 | 申请日 | 公开日 | 申请人 | 专利标题

---

---

CA710277A|1965-05-25|E. Barnhart Charles|Feedstuff for inhibiting anemia in swine|

---

US2946722A|1959-07-21|1960-07-26|Walter H Hoffman|Composition and method for treating poultry|

---

US3259500A|1961-10-11|1966-07-05|Kentucky Res Foundation|Method for inhibiting anemia in young pigs by feeding the sow an iron agent|

---

US3558778A|1966-10-27|1971-01-26|Dow Chemical Co|Methods and compositions for use in animal husbandry|

---

US3491187A|1967-02-27|1970-01-20|Diamond Shamrock Corp|Iron salt-antibiotic compositions and use thereof|

---

US4053592A|1973-03-16|1977-10-11|Abbott Laboratories|Animal growth promotant|

---

US4171379A|1978-04-05|1979-10-16|Ralston Purina Company|Feed supplement for baby pigs|

---

US4504471A|1982-09-27|1985-03-12|Fujisawa Pharmaceutical Co., Ltd.|Animal growth promotant and method of use for animal growth|

---

AT197136T|1990-05-02|2000-11-15|Grace W R & Co|POLYMER COMPOSITIONS WITH OXYGEN INTERCEPTING COMPOUNDS|

---

US6024979A|1995-06-06|2000-02-15|Solvay Animal Health, Inc.|Oral veterinary composition containing a fluoroquinolone antibacterial agent possessing superior absorption properties and an extended duration of therapeutic antimicrobial blood levels, and a method of treating a microbial infection in a ruminant|

---

AU4134697A|1996-09-06|1998-03-26|Otsuka Pharmaceutical Co., Ltd.|Composition, antimicrobial agent, infection preventive, and food against |

---

US5879697A|1997-04-30|1999-03-09|Schneider Usa Inc|Drug-releasing coatings for medical devices|

---

KR20010013377A|1997-06-04|2001-02-26|데이비드 엠 모이어|Mild, leave-on antimicrobial compositions|

---

US6139879A|1997-06-25|2000-10-31|Foliar Nutrients, Inc.|Fungicidal and bactericidal compositions for plants containing compounds in the form of heavy metal chelates|

---

US6197815B1|1998-03-18|2001-03-06|J.H. Biotech, Inc.|Amino acid chelates to reduce still births and increase birth weight in non-human animals|

---

US6773737B1|1998-11-19|2004-08-10|The Procter & Gamble Company|Microorganism reduction methods and compositions for food|

---

US7143721B2|1999-03-10|2006-12-05|Ridley Block Operations, Inc.|Reducing ungulate pressure on post-burn areas in mosaic burns|

---

US7247338B2|2001-05-16|2007-07-24|Regents Of The University Of Minnesota|Coating medical devices|

---

US7431939B1|2000-09-28|2008-10-07|Mississippi State University|Inhibition of systemic infections in humans and vertebrates by dietary fibers|

---

AU2002246679B2|2000-12-18|2006-11-16|Probiohealth Llc|Probiotic compounds derived from lactobacillus casei strain KE01|

---

US6626397B2|2001-07-02|2003-09-30|Elsbit Technologies Ltd.|Autonomous flying wing|

---

US6669980B2|2001-09-18|2003-12-30|Scimed Life Systems, Inc.|Method for spray-coating medical devices|

---

SE0103695D0|2001-11-07|2001-11-07|Thomas Boren|A novel non-antibiotic strategy against OGIP infections based on a cereal product|

---

CN1245093C|2003-08-12|2006-03-15|石家庄市科星动物保健品有限公司|Prefabricated amino chelate mixed feed|

---

US7258875B2|2003-12-04|2007-08-21|Chiou Consulting, Inc.|Compositions and methods for topical treatment of skin infection|

---

EP2497478A3|2003-12-05|2012-11-07|Children's Hospital Medical Center|Oligosaccaride compositions and use thereof in the treatment of infection|

---

DE10356975A1|2003-12-05|2005-07-07|Manuel Dipl.-Kfm. Engels|Between towering, load-bearing building suspended building|

---

JP2005325080A|2004-05-17|2005-11-24|Daiichi Fine Chemical Co Ltd|Composition containing calcium pantothenate and vitamins|

---

GB0422052D0|2004-10-04|2004-11-03|Dansico As|Enzymes|

---

GB0423681D0|2004-10-26|2004-11-24|Sec Dep For Environment Food &|Vaccine and nucleic acids|

---

US20060134227A1|2004-12-22|2006-06-22|Bortz Jonathan D|Compositions including iron|

---

EP1876909A2|2005-03-11|2008-01-16|Archer-Daniels-Midland Company|Compositions and methods providing rumen bypass protein in ruminant diets|

---

US20090081315A1|2005-04-08|2009-03-26|Snow Brand Milk Products Co., Ltd.|Iron composition containing milk protein|

---

JP2006321776A|2005-05-18|2006-11-30|Kyuuken:Kk|Antibacterial mildewproofing agent of metal complex coordinated with amino acid and its secondary product|

---

US20070269495A1|2006-05-18|2007-11-22|Ashmead H Dewayne|Compositions and methods for enhancing mineral levels in animals with reduced environmental impact|

---

US20100249058A1|2006-10-17|2010-09-30|Idemitsu Kosan Co., Ltd.|Feed additive and feed|

---

US20080194679A1|2007-02-14|2008-08-14|Ashmead H Dewayne|Compositions and methods for increasing metabolic activity in animal tissue|

---

WO2008105983A1|2007-02-27|2008-09-04|Albion International, Inc.|Mineral absorption from the stomach|

---

WO2009089493A2|2008-01-11|2009-07-16|Albion International, Inc.|Nitrate amino acid chelates|

---

CN102065696B|2008-05-01|2013-08-28|万灵杀菌消毒剂股份有限公司|Versatile disinfectant|

---

AU2009313998B2|2008-11-12|2016-06-16|Synedgen, Inc.|Chitosan derivatives to treat animals or optimize animal health|

---

US8273514B2|2009-05-22|2012-09-25|Xerox Corporation|Interfacial layer and coating solution for forming the same|

---

TW201103439A|2009-06-08|2011-02-01|Idemitsu Kosan Co|Coccidiosis control agent and feed containing same|

---

US8178709B2|2009-07-21|2012-05-15|Biolink Life Sciences, Inc.|Iron preparation suitable for pharmaceutical formulation and process for the preparation thereof|

---

WO2011086569A1|2010-01-18|2011-07-21|Concept Medical Research Private Limited|Method and system for coating insertable medical devices|

---

WO2012034032A2|2010-09-10|2012-03-15|The Board Of Regents Of Unbiversity Of Texas System|Antimicrobial solutions|

---

CA2820706C|2010-12-03|2018-05-22|Ms Technologies, Llc|Optimized expression of glyphosate resistance encoding nucleic acid molecules in plant cells|

---

US8741375B2|2011-06-07|2014-06-03|Zinpro Corporation|Mixed amino acid metal salt complexes|

---

ITMI20111033A1|2011-06-08|2012-12-09|Giuliano Leonardi|PROCEDURE FOR THE PREPARATION OF A COMPOSITION INCLUDING METAL ORGANIC OIL COMPLEX|

---

JP6266510B2|2011-06-08|2018-01-24|キレーション パートナーズ インコーポレイテッド|Metal chelate compositions and methods for controlling the growth or activity of living cells or organisms|

---

US20130022706A1|2011-07-20|2013-01-24|Inco Digestive, Llc|Animal Feed Additive|

---

EP3569230A1|2012-02-16|2019-11-20|Akeso Biomedical, Inc.|Reduction of gastrointestinal tract colonisation by campylobacter|US8802868B2|2010-03-25|2014-08-12|Vertex Pharmaceuticals Incorporated|Solid forms of -1-N--1H-Indol-5-yl)-Cyclopropanecarboxamide|

---

WO2011133751A2|2010-04-22|2011-10-27|Vertex Pharmaceuticals Incorporated|Process of producing cycloalkylcarboxamido-indole compounds|

---

US9768463B2|2012-07-27|2017-09-19|Lockheed Martin Advanced Energy Storage, Llc|Aqueous redox flow batteries comprising metal ligand coordination compounds|

---

US9382274B2|2012-07-27|2016-07-05|Lockheed Martin Advanced Energy Storage, Llc|Aqueous redox flow batteries featuring improved cell design characteristics|

---

US10164284B2|2012-07-27|2018-12-25|Lockheed Martin Energy, Llc|Aqueous redox flow batteries featuring improved cell design characteristics|

---

DK2948129T3|2013-01-25|2018-01-29|Wintermute Biomedical Inc|Therapeutic compounds|

---

CN110840847A|2014-04-15|2020-02-28|沃泰克斯药物股份有限公司|Pharmaceutical compositions for the treatment of cystic fibrosis transmembrane conductance regulator mediated diseases|

---

US10555531B2|2015-08-11|2020-02-11|Akeso Biomedical, Inc.|Biofilm inhibiting compositions enhancing weight gain in livestock|

---

WO2016086163A1|2014-11-26|2016-06-02|Lockheed Martin Advanced Energy Storage, Llc|Metal complexes of substituted catecholates and redox flow batteries containing the same|

---

US10253051B2|2015-03-16|2019-04-09|Lockheed Martin Energy, Llc|Preparation of titanium catecholate complexes in aqueous solution using titanium tetrachloride or titanium oxychloride|

---

US10644342B2|2016-03-03|2020-05-05|Lockheed Martin Energy, Llc|Coordination complexes containing monosulfonated catecholate ligands and methods for producing the same|

---

US10316047B2|2016-03-03|2019-06-11|Lockheed Martin Energy, Llc|Processes for forming coordination complexes containing monosulfonated catecholate ligands|

---

WO2017173230A1|2016-04-01|2017-10-05|Texas State University|Compositions and methods for dispersing biofilms|

---

US9938308B2|2016-04-07|2018-04-10|Lockheed Martin Energy, Llc|Coordination compounds having redox non-innocent ligands and flow batteries containing the same|

---

US10377687B2|2016-07-26|2019-08-13|Lockheed Martin Energy, Llc|Processes for forming titanium catechol complexes|

---

US10343964B2|2016-07-26|2019-07-09|Lockheed Martin Energy, Llc|Processes for forming titanium catechol complexes|

---

DK3436054T3|2016-09-13|2019-11-11|Allergan Inc|STABILIZED NON-PROTEIN CLOSTRID TOXIN COMPOSITIONS|

---

US10065977B2|2016-10-19|2018-09-04|Lockheed Martin Advanced Energy Storage, Llc|Concerted processes for forming 1,2,4-trihydroxybenzene from hydroquinone|

---

US10930937B2|2016-11-23|2021-02-23|Lockheed Martin Energy, Llc|Flow batteries incorporating active materials containing doubly bridged aromatic groups|

---

RU2671387C2|2016-12-12|2018-10-30|Федеральное государственное бюджетное научное учреждение "Краснодарский научый центр по зоотехнии и ветеринарии" |Means for disinfection of hatchery eggs and increasing their hatchability|

---

US10497958B2|2016-12-14|2019-12-03|Lockheed Martin Energy, Llc|Coordinatively unsaturated titanium catecholate complexes and processes associated therewith|

---

US10741864B2|2016-12-30|2020-08-11|Lockheed Martin Energy, Llc|Aqueous methods for forming titanium catecholate complexes and associated compositions|

---

CA3052047A1|2017-02-13|2018-08-16|Wintermute Biomedical, Inc.|Anti-pathogenic therapeutic compositions|

---

US10320023B2|2017-02-16|2019-06-11|Lockheed Martin Energy, Llc|Neat methods for forming titanium catecholate complexes and associated compositions|

---

ES2877900T3|2017-06-15|2021-11-17|Nikken Lease Kogyo Co Ltd|Anesthesia Maintenance System and Anesthesia Maintenance Method for Fish and Shellfish|

---

CN108094410B|2018-01-26|2021-08-31|山东大学齐鲁医院|Skin deep hypothermia protective agent and skin preservation method|

---

CN108748532A|2018-06-08|2018-11-06|阜南县永盛工艺品有限公司|A kind of dyeing treatment of levant cotton exocarp|

---

CN108651758A|2018-07-25|2018-10-16|石阡县毅峰农牧开发有限公司|One kind chicken feed containing phytase and preparation method thereof|

---

CN109200340A|2018-08-18|2019-01-15|费宇奇|A kind of preparation method of antibacterial inertia medical catheter|

---

US20200078149A1|2018-09-08|2020-03-12|Ali Mohammad Saghiri|Expandable Floss with Specialized Tray|

---

CN110935057A|2018-09-21|2020-03-31|天津大学|Application of dopamine-based tissue adhesive in antibacterial biomedical materials|

---

KR20210108942A|2018-10-01|2021-09-03|윈터뮤트 바이오메디컬 인코포레이티드|therapeutic composition|

---

US10927397B2|2018-10-16|2021-02-23|Sterilex, Llc|Compositions, devices and methods for detecting biofilms|

---

RU2689163C1|2018-11-06|2019-05-24|федеральное государственное бюджетное образовательное учреждение высшего образования "Хакасский государственный университет им. Н.Ф. Катанова" |Method of lymphatic therapy in pyelonephritis in dogs|

---

WO2020097213A2|2018-11-07|2020-05-14|The Regents Of The University Of Michigan|Compositions and methods for production of cysteine|

---

CN109512581A|2018-11-11|2019-03-26|南通强生石墨烯科技有限公司|A kind of graphene sanitary napkin|

---

WO2020252003A1|2019-06-13|2020-12-17|Hollister Incorporated|Reusable urinary catheter products|

---

CN110235812B|2019-06-18|2021-12-21|中国水产科学研究院东海水产研究所|Mixed culture method of verasper variegates and turbot|

---

CN110393159B|2019-08-19|2021-10-26|东北农业大学|Egg cleaning and disinfecting method based on ultrasonic-micro nano bubble technology|

---

RU2758056C2|2019-11-21|2021-10-26|федеральное государственное бюджетное образовательное учреждение высшего образования "Ставропольский государственный аграрный университет"|Drug for the treatment of otitis of bacterial and fungal etiology in dogs|

---

CN111387135B|2020-03-13|2021-10-29|北京家禽育种有限公司|Method for systematically monitoring and purifying pseudomonas aeruginosa in hatching process|

法律状态:
2019-11-05| B06U| Preliminary requirement: requests with searches performed by other patent offices: procedure suspended [chapter 6.21 patent gazette]|

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2021-10-05| B15K| Others concerning applications: alteration of classification|Free format text: AS CLASSIFICACOES ANTERIORES ERAM: A23L 1/304 , A01N 1/00 , A01N 59/16 , A23K 1/175 , A61K 35/12 , A61K 35/36 , A61K 35/37 , C09D 5/14 , A01N 37/10 , A01N 43/40 Ipc: A01K 14/00 (2006.01), A01K 43/00 (2006.01), A01N 3 |

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2021-11-09| B09A| Decision: intention to grant [chapter 9.1 patent gazette]|

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2022-01-18| B16A| Patent or certificate of addition of invention granted [chapter 16.1 patent gazette]|Free format text: PRAZO DE VALIDADE: 20 (VINTE) ANOS CONTADOS A PARTIR DE 11/08/2015, OBSERVADAS AS CONDICOES LEGAIS. |

优先权:

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申请号 | 申请日 | 专利标题

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US201462036790P| true| 2014-08-13|2014-08-13|

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US62/036,790|2014-08-13|

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US201562137630P| true| 2015-03-24|2015-03-24|

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US62/137,630|2015-03-24|

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US201562138499P| true| 2015-03-26|2015-03-26|

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US62/138,499|2015-03-26|

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US201562171081P| true| 2015-06-04|2015-06-04|

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US62/171,081|2015-06-04|

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US201562188183P| true| 2015-07-02|2015-07-02|

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US62/188,183|2015-07-02|

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PCT/US2015/044603|WO2016025448A2|2014-08-13|2015-08-11|Antimicrobial compounds and compositions, and uses thereof|

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