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    • 专利摘要:
    Bacterial strain of Streptomyces althioticus that produces desertomycin and desertomycin produced by it. The invention provides a new bacterial strain of Streptomyces althioticus isolated from its natural environment and deposited in the Spanish Type Culture Collection under accession number (CECT 9651), which is capable of efficiently producing antibiotic and cytotoxic compounds of the family by fermentation. of desertomycins. Specifically, this strain is a producer of a new desertomycin, designated in the present invention as desertomycin G, which has antibiotic activities against clinical and cytotoxic pathogens against tumor cell lines, preferably human, preferably from the pancreas, breast and colon. (Machine-translation by Google Translate, not legally binding)
    公开号:ES2752040A1
    申请号:ES201800220
    申请日:2018-10-02
    公开日:2020-04-02
    发明作者:Brana Alfredo Javier Fernandez；Vizcaino Aida Sarmiento；Diaz Luis Arsenio Garcia；Blanco María Gloria Blanco；Mendez Marina Salmon；Guerra Luis Otero；Gutierrez Juan José Palacios；Suarez Jonathan Fernandez；Benitez José Fernando Reyes；Moreno De Barreda Ignacio Perez-Victoria；Serrano Jesús Melchor Martin；Munarriz Yamina Mohamedi；Lopez Tania Fontanil；Miguel Santiago Cal
    申请人:Hospital Univ Central De Asturias；Hospital Univesitario De Cabuenes；Universidad de Oviedo；
    IPC主号:
    专利说明:

    [0001]
    [0002] Bacterial strain of streptomyces althioticus producing desertomycin and desertomycin produced by it
    [0003]
    [0004] Technical sector
    [0005]
    [0006] The present invention falls within the pharmaceutical field, and specifically refers to new natural products with application in the treatment of bacterial infections and in oncology, which are obtained by fermentation of a bacterial strain that produces antibiotic and / or antitumor compounds of the family of desertomycins. The invention also relates to a new desertomycin, desertomycin G, to its preparation procedure and to pharmaceutical compositions comprising it, which are useful for the treatment and / or prevention of bacterial infections caused by pathogenic Gram-positive bacteria, such as Mycobacterium tuberculosis and Staphylococus aureus, preferably resistant to clinically relevant antibiotics; as well as the treatment and / or prevention of tumors, preferably of the breast and colon in humans.
    [0007]
    [0008] Background of the Invention
    [0009]
    [0010] Desertomycins are natural products of pharmacological interest due to their various biological activities. Structurally, they belong to the family of marginolactones, polyketide aminopolyol containing a macrolactone ring whose biosynthesis is under active investigation (Hong et al., 2016, Angew Chem Int Ed Engl. 55: 1118-23.). Since the discovery of desertomycin A, first described as an antibiotic against bacteria and fungi (Uri et al., 1958, Nature. 182: 401), other compounds in the family have since been identified as desertomycin B and D ( Bortolo et al., 1992, J Antibiot 45: 1016-9), E (Ivanova et al., 1997, Prep Biochem Biotechnol. 27: 19-38) and F (Dinya et al., 1996, Rapid Commun. Mass Spectrom ., 10, 1439-1448). Desertomycins are mainly produced by species of the genus Streptomyces isolated from soils, such as Streptomyces macronensis and Streptomyces flavofungini (Dolak et a /. 1982, J Antibiot, 36: 139) and also by Streptoverticillium baldaccii (Mayer and Thiericke, 1993 J. Chem. Soc., Perkin Trans. 1, 2525-2531).
    [0011]
    [0012] The marine environment has become a primary resource in the search and discovery of new natural products from marine actinomycetes (Ward and Bora, 2006, Curr Opin Microbiol 9: 279-286). Seaweeds (macroalgae) remain a relatively unexplored source in the search for actinomycetes that produce bioactive compounds of pharmacological interest. Previous works describe the isolation of actinomycetes of the genus Streptomyces from marine algae in coastal ecosystems of temperate and cold waters of the North Atlantic Ocean, particularly on the Atlantic and Mediterranean coasts of the Iberian Peninsula (Genilloud et al., 1994, Microbiology 10: 413-422) and the Kiel Fjord in the Baltic Sea (Staufenberger et al. 2008, FEMS Microbiol Ecol 64: 65-77; Wiese et al., 2009, Mar Biotechnol 11: 287-300). Recent research in the Cantabrian Sea (Bay of Biscay), Northeast Atlantic, has revealed the presence of bioactive Streptomyces species associated with marine macroalgae (Braña et a / 2015, Microb Ecol 69: 512-24; Sarmiento-Vizcaíno et al. 2016 , Microb Ecol. 71: 375-86).
    [0013]
    [0014] Due to the growing appearance in the clinical setting of pathogenic bacteria resistant to current antibiotics, it is necessary to obtain new antibiotics with bio-medical potential in the treatment and prevention of infectious diseases caused by pathogenic bacteria. Furthermore, there is a growing need for new antitumor agents, with improved activities, with fewer undesirable side effects and with higher selectivity, compared to the drugs currently in use. In the fight against cancer, it is necessary to obtain new antitumor drugs with improved pharmacological properties. That's it It is necessary to have simple, short and inexpensive procedures to obtain said compounds with antibacterial and / or antitumor activity.
    [0015]
    [0016] Explanation of the invention
    [0017]
    [0018] The present invention provides a new bacterial strain of Streptomyces althioticus that has been isolated from its natural environment and deposited in the Spanish Type Culture Collection under accession number (CECT 9651), which is capable of efficiently producing antibiotic compounds by fermentation and cytotoxic agents from the desertomycin family. Specifically, this strain is a producer of a new desertomycin, designated in the present invention as desertomycin G, which exhibits antibiotic activities against clinical pathogens and cytotoxic against tumor cell lines, preferably human.
    [0019]
    [0020] The inventors of the present invention have isolated said strain, Streptomyces althioticus MSM3, from the intertidal ecosystems of the Cantabrian Sea. This strain was isolated from the intertidal algae Ulva sp., Collected in Pedreña (Cantabria) and was studied and subsequently identified. Therefore, this strain is described in the present invention, a procedure to obtain desertomycins by fermentation from it and a new natural product of the desertomycin family with antibacterial activity in vivo against Gram-positive clinical pathogenic bacteria, such as example but not limited to, Mycobacterium tuberculosis and Staphylococcus aureus. Desertomycin G also presents cytotoxic activity against human tumor lines, such as, but not limited to, lines of breast adenocarcinoma and colon carcinoma.
    [0021]
    [0022] The inventors determined the culture conditions of the strain for the production of the new desertomycin, subsequently purified it, proceeded with its structural elucidation and tested its antibiotic activity against a variety of Gram-positive and Gram-negative clinical pathogens, and cytotoxic against to various human tumor lines.
    [0023]
    [0024] The present invention thus represents a solution to the need for new compounds with biomedical potential in the treatment or prevention of infectious diseases caused by pathogenic Gram-positive bacteria, as well as new antitumor compounds. Likewise, it represents a solution to the need for simple, short and inexpensive procedures to obtain said compound with antibiotic and antitumor activity, since the procedure described in the present invention allows the said compound to be produced by fermentation with an actinobacterium, in instead of by chemical synthesis, a more complex, long and expensive process. In this sense, in biotechnological processes that involve obtaining structurally complex natural products, such as desertomycins, production by fermentation using the producing microorganism is the preferred procedure, since it is simpler, shorter and cheaper than the organic synthesis procedure.
    [0025]
    [0026] Thus, in a first aspect, the present invention relates to a bacterial strain of Streptomyces althioticus MSM3 deposited on May 16, 2018 in the Spanish Type Culture Collection under the accession number CECT 9651. From now on reference will be made to this bacterial strain as "strain of the invention" or "bacterial strain of the invention".
    [0027]
    [0028] Another aspect of the invention relates to a supernatant or extract of a culture of the bacterial strain of the invention. This supernatant will be referred to as "supernatant of the invention", and comprises at least one compound of general formula (I) described below.
    [0029] The supernatant of the invention comprising at least one compound of formula (I) can be obtained by cultivating the strain of the invention in the presence of a suitable culture medium and under fermentation conditions. Said fermentation conditions and said medium culture are preferably those described below in the process of the invention. Subsequently, the bacterial culture can be centrifuged, by any method known to those skilled in the art for this purpose, and the sediment deposited as a consequence of this centrifugation step can be removed, thus obtaining the supernatant of the invention that it comprises at least one compound of formula (I) and other metabolites produced and secreted by the strain of the invention in culture.
    [0030] Another aspect of the invention relates to the use of the bacterial strain of the invention for the production of compounds of general formula (I):
    [0031]
    [0032]
    [0033]
    [0034] or any of its salts or tautomers,
    [0035] where,
    [0036] R 1 is selected from hydrogen, C 1 -C 6 alkyl substituted or unsubstituted alkenyl C2 - C6 substituted or unsubstituted alkynyl , C2 - C6 substituted or unsubstituted, C1 - C6 alkylacyl substituted or unsubstituted , substituted or unsubstituted benzyl or substituted or unsubstituted benzoyl.
    [0037] R2 and R3 are independently selected from hydrogen, COR a, COOR a, alkyl C1 - C6 substituted or unsubstituted alkyl , C2 - C6 substituted or unsubstituted alkynyl or C2 - C6 substituted or unsubstituted, where IR , is selected from hydrogen, C1-C6 substituted or unsubstituted alkyl , C2 - C6 substituted or unsubstituted alkynyl, C2 - C6 substituted or unsubstituted aryl , substituted or unsubstituted or a heterocyclic group substituted or unsubstituted .
    [0038] Another aspect of the invention refers to a process for obtaining a compound of general formula (I) described above, hereinafter "process of the invention", comprising:
    [0039] to. cultivating the bacterial strain of the invention in a culture medium under fermentation conditions, and
    [0040] b. purify the compound of general formula (I) produced by the cultivated strain of step (a).
    [0041]
    [0042] The cultivation of step (a) of the process of the invention can be carried out, for example, but without limitation, by inoculating the strain or spores of the strain in an appropriate culture medium. Those skilled in the art will recognize the bacterial culture media that can be used in this stage of the process of the invention. The "culture medium" is a suitable nutrient medium, that is, it comprises the nutrients necessary for the maintenance and in vitro growth of the strain of the invention, for the development of its fermenting activity and, therefore, for the production of the compounds of formula (I) described above. Said culture medium can be liquid, solid or semi-solid. In order for the strain of the invention to grow properly in the culture medium, it must meet a series of conditions such as temperature, agitation, degree of humidity, light / dark and adequate oxygen pressure, as well as a correct degree of acidity or alkalinity. (pH). Likewise, the culture medium must be free of all contaminating microorganisms.
    [0043]
    [0044] Thus, the cultivation of step (a) takes place in a suitable nutrient medium comprising, for example but not limited to, agar or gelatin or albumin, carbon sources (eg glucose, sucrose or mannitol), nitrogen sources ( for example, peptones), sulfur, phosphorous, sources of vitamins, amino acids and hormones and / or growth factors (for example, meat extract or yeast extract), MOPS, inorganic salts (for example, calcium in the form of CaCl2, magnesium, manganese, sodium or potassium), hydrogen ions, etc. In a preferred embodiment, the culture medium used in the process of the invention is a culture medium comprising nutrients and physico-chemical properties similar to those of the marine environment, more preferably the medium is marine R5A
    [0045]
    [0046] The strain of the invention can be grown, for example, but not limited to, by shake flask culture, and small or large-scale fermentation (including continuous, batch or batch-fed, feed-batch, or in solid state) carried out in a laboratory or industrial bioreactor in a suitable medium and under conditions that allow expressing and / or isolating the compounds of formula (I) described above. The compounds of the invention are secreted, together with other metabolites or compounds, in the nutrient medium, and these can be recovered directly from the medium.
    [0047]
    [0048] The person skilled in the art will recognize the suitable fermentation conditions to be applied in step (a) of the process of the invention. Preferably, said conditions comprise incubation with shaking, more preferably in an orbital shaker, of the culture of the invention for between 2 and 7 days, preferably between 3 and 5 days, more preferably for 4 days; at an essentially constant temperature of between 25 and 30 ° C, preferably between 27 and 29 ° C, more preferably at 28 ° C; at between 200 and 300 rpm, preferably between 250 and 270 rpm, more preferably at 250 rpm and at a constant pH of between 6.0 and 7.5, preferably between 6.0 and 7.0, more preferably at 6.7.
    [0049]
    [0050] After cultivation of step (a) of the process of the invention, an additional centrifugation step can take place after which the sediments are discarded and the supernatants are selected, thereby obtaining the supernatant of the invention. Said supernatants can be subsequently filtered in step (b) of the process of the invention and can then be subjected to an extraction procedure, such as, but not limited to, solid phase extraction, in order to elute the compound of the invention present therein.
    [0051] The compound of the invention secreted into the culture medium by the strain of the invention can be recovered from the medium using procedures known in the art, for example by Conventional procedures including, but not limited to, centrifugation, filtration, extraction, spray drying, evaporation, and / or precipitation.
    [0052]
    [0053] The compound of the invention produced by the cultured strain can be purified by a variety of procedures known in the art including, but not limited to, chromatography (eg, HPLC, ion exchange, affinity, hydrophobic, chromato focus, and molecular exclusion), procedures electrophoretic (eg, preparative isoelectric focusing), differential solubility (eg, ammonium sulfate precipitation), SDS-PAGE, precipitation, or extraction, to obtain the compound of the invention substantially pure.
    [0054]
    [0055] The compound of the invention produced by the strain in culture can be detected using procedures known in the art. These detection procedures may include, for example, but are not limited to, UPLC, UV, NMR, mass spectrometry, or the like.
    [0056]
    [0057] Another aspect of the invention relates to a compound of general formula (I) or to any of its salts or tautomers (hereinafter "compound of formula (I) of the invention", "compound of the invention" or "desertomycin G "):
    [0058]
    [0059]
    [0060]
    [0061]
    [0062] where,
    [0063]
    [0064] R1 substituted 6 is selected from hydrogen, C 1 -C alkyl or unsubstituted alkyl , C2 - C6 substituted or unsubstituted alkynyl , C2 - C6 substituted or unsubstituted, C1 - C6 alkylacyl substituted or unsubstituted, substituted or unsubstituted benzyl and substituted or unsubstituted benzoyl.
    [0065]
    [0066] More preferably the group R1 is selected from hydrogen, substituted or unsubstituted methyl, substituted or unsubstituted ethyl, substituted or unsubstituted n-propyl, substituted or unsubstituted iso-propyl, substituted or unsubstituted n-butyl, tert-butyl substituted or unsubstituted, substituted or unsubstituted vinyl, substituted or unsubstituted allyl, substituted or unsubstituted ethynyl, substituted or unsubstituted acetyl, substituted or unsubstituted benzyl, and substituted or unsubstituted benzoyl.
    [0067] Even more preferably, the group R 1 is selected from hydrogen, acetyl, benzyl, and benzoyl
    [0068]
    [0069] In another more preferred embodiment, the group R 1 is hydrogen.
    [0070]
    [0071] R 2 and R 3 are independently selected from hydrogen, COR a, COOR a, C 1 -C 6 substituted or unsubstituted alkyl , C2 - C6 substituted or unsubstituted alkynyl C2 - C6 substituted or unsubstituted.
    [0072]
    [0073] R a is selected from hydrogen, alkyl Cr C6 substituted or unsubstituted alkenyl C2 - C6 substituted or unsubstituted alkynyl C2 - C6 substituted or unsubstituted aryl, substituted or unsubstituted or heterocyclic group substituted or unsubstituted .
    [0074]
    [0075] More preferably R 2 and R 3 are independently selected from hydrogen, COR a , COOR a, and substituted or unsubstituted C 1 -C 6 alkyl, where Ra is selected from hydrogen and substituted or unsubstituted C 1 -C 6 alkyl. Even more preferred substituents Ra are methyl, ethyl, n-propyl, isopropyl and butyl, including n-butyl, tert-butyl, sec-butyl and iso-butyl.
    [0076]
    [0077] Even more preferred is the compound in which R 2 and R 3 are hydrogen.
    [0078]
    [0079] In particular, the present invention provides, among others, the compound with the following formula (II):
    [0080]
    [0081]
    [0082]
    [0083]
    [0084] Another aspect of the invention relates to a composition, hereinafter "composition of the invention", comprising the strain of the invention, the supernatant of the invention or the compound of formula (I) of the invention. Said composition may be a pharmaceutical or cosmetic composition, comprising the strain of the invention, the supernatant of the invention or the compound of formula (I) of the invention, and a pharmaceutically or cosmetically acceptable excipient or vehicle. Preferably, the composition of the invention comprises the strain of the invention, the supernatant of the invention or the compound of formula (I) of the invention in a therapeutically effective amount.
    [0085] By "therapeutically effective amount" is meant the amount of the strain of the invention, supernatant of the invention or compound of the invention, that when administered to the subject to treat and / or prevent a bacterial infection, or a tumor process, produces the effect wanted. Said desired effect may be, for example but not limited to, eliminating or reducing the number of bacteria causing the infection or the number of tumor cells. The therapeutically effective amount may vary depending on a variety of factors, for example but not limited to, the type of tumor or infection and its severity, as well as age, weight, sex, physical condition, responsiveness, or tolerance, etc. of the individual to whom the composition of the invention is to be administered.
    [0086]
    [0087] The pharmaceutically or cosmetically acceptable excipients and carriers that can be used in the composition of the invention are those known to those skilled in the art.
    [0088]
    [0089] The term "excipient" refers to a substance that aids in the absorption of the elements of the composition of the invention, stabilizes said elements, activates or helps the preparation of the composition in the sense of giving it consistency. Thus, the excipients could have the function of keeping the ingredients together, as for example is the case of starches, sugars or cellulose, the sweetening function, the coloring function, the protection function of the composition, such as, for example, to isolate it from air and / or humidity, the filling function of a tablet, capsule or any other form of presentation, such as, for example, the case of dibasic calcium phosphate, the disintegrating function to facilitate the dissolution of the components and their absorption, without excluding other types of excipients not mentioned in this paragraph.
    [0090]
    [0091] The "pharmacologically acceptable vehicle", like the excipient, is a substance or combination of substances that is used in the composition to dilute any of the components included in it to a certain volume or weight. The term "vehicle" refers to a diluent, adjuvant, excipient, or carrier with which the composition of the invention is to be administered; obviously, said vehicle must be compatible with said composition. Pharmaceutically acceptable carriers can be, but are not limited to, solids, liquids, solvents, or surfactants. Examples of vehicles are, but are not limited to, water, oils, or surfactants, including those of petroleum, animal, vegetable, or synthetic origin, such as, for example, and without limitation, peanut oil, soybean oil, mineral oil, sesame oil , castor oils, polysorbates, sorbitan esters, ether sulfates, sulfates, betaines, glucosides, maltosides, fatty alcohols, nonoxynols, poloxamers, polyoxyethylenes, polyethylene glycols, dextrose, glycerol, digitonin and the like. The pharmacologically acceptable vehicle is an inert substance or of action analogous to any of the elements included in the composition of the present invention. The function of the vehicle is to facilitate the incorporation of other elements, to allow a better dosage and administration or to give consistency and shape to the composition. When the presentation form is liquid, the pharmacologically acceptable vehicle is the diluent.
    [0092]
    [0093] The composition of the present invention can be formulated for administration to an animal, preferably a mammal, including man, in a variety of ways known in the art. Examples of preparations include any solid composition (tablets, pills, capsules, powders, granules, sticks, pencils, vaporizers, sprays, etc.), semi-solid (ointment, cream, ointment, gel, hydrogel, foam, lotion, soap, jelly, gelatin, etc.) or liquid (aqueous or non-aqueous solutions, hydroalcoholic or hydro-glycolic solutions, suspensions, emulsions, syrups, anhydrous compositions, aqueous dispersions, oils, milk, balms, liniments, serums, etc.) for oral administration, topical or parenteral. The composition of the present invention may also be in the form of sustained release formulations or any other conventional release system. The term "sustained release" is used in the conventional sense referring to a vehicle system of a compound that provides the gradual release of said compound over a period of time and preferably, but not necessarily, with relatively constant levels of release of the compound over a period of time. Illustrative examples of sustained release vehicles or systems include, but are not limited to, liposomes, mixed liposomes, oleosomes, niosomes, etosomes, millycapsules, microcapsules, nanocapsules, sponges, cyclodextrins, vesicles, micelles, mixed surfactant micelles, phospholipidotensive mixed micelles , milliespheres, microspheres, nanospheres, lipospheres, microemulsions, nanoemulsions, miniparticles, milliparticles, microparticles, nanoparticles, solid lipid nanoparticles, nanostructured lipid supports polymeric materials, biodegradable or non-biodegradable patches or implants, or biodegradable microparticles, for example.
    [0094]
    [0095] Such compositions and / or their formulations can be administered in a variety of ways, including, but not limited to, parenteral, intraperitoneal, intravenous, intradermal, epidural, intraspinal, intrastromal, intraarticular, intrasynovial, intrathecal, intralesional, intraarterial, intracardiac, intramuscular, intranasal, intracranial, cutaneous or subcutaneous, intraorbital, intracapsular, topical, ophthalmological or ocular, using transdermal patches or rectally or vaginally, by administering a suppository or encapsulated, percutaneous, nasal spray, surgical implant, internal surgical paint, pump infusion or catheter.
    [0096]
    [0097] The compositions of the present invention are suitable for application by medical devices that allow the release of the active ingredient in concentrations suitable for the treatment of bacterial infections or tumor processes. These devices should preferably be suitable for the administration of the active ingredient locally, allowing the treatment to act on the affected area and not to disperse. Devices can, for example, but not be limited to, carry or be coated with the active ingredient.
    [0098] In a preferred embodiment, the composition of the invention further comprises another antibacterial and / or antitumor agent, more preferably said antibacterial agent is another antibiotic.
    [0099]
    [0100] "Antibacterial agent" is understood to be a chemical substance, produced synthetically or naturally (synthesized for example by fungi or bacteria), that inhibits the growth (bacteriostatic) or kills (bactericidal) the bacteria. The antibacterial agent may be, for example but not limited to, an antibiotic, an efflux pump inhibitor, or a bacterial membrane permeabilizing agent.
    [0101]
    [0102] The antibiotics to which the present invention refers are compounds that preferably do not compromise the viability and survival of the strain of the invention. Examples of antibiotics that can be included in the composition of the invention are, but are not limited to, arnikacin, gentamicin, kanamycin, neomycin, netilmycin, streptomycin, tobramycin, paromomycin, geldanamicin, herbimycin, loracarbef, ertapenem, doripenem, imipenem / cilastatin, cefadroxil, cefazolin, cephalothin, cephalexin, cefaclor, cefamandol, cefoxitin, cefuroxime, cefixime, cefdinir, cefditerin, cefoperazine, cefpoxime, cefpoxime, cefpoxime, cefibuziphetin , erythromycin, roxithromycin, troleandomycin, telithromycin, spectinomycin, aztreonam, amoxicillin, ampicillin, azlocillin, carbenicillin, cloxacillin, dicloxacillin, mexillin, methylline, nazillin, penicillin , levofloxacin, lomefloxacin, mo Xifloxacin, Norfloxacin, Ofloxacin, Trovafloxacin, Grepafloxacin, Sparfloxacin, Temafloxacin, Mafenide, Sulfonamidocrisoidine, Sulfacetamilacin, Sulphametizine, Sulphamethacim, Sulfanilimidae, Trimethoim, , arsphenamine, chloramphenicol, clindamycin, lincomycin, ethambutol, phosphomycin, fusidic acid, furazolidone, isoniazid, linezolid, metronidazole, mupirocin, nitrofurantoin, platensimycin, pyrazinamide, quinupristin / dalfopristin, rifampin, thiamphenicol, tinidazole, dapsone, and clofazimine, as well as other nargenicins, including desertomycins other than those described in the present invention.
    [0103] The composition of the present invention may alternatively or additionally include an antifungal or antifungal agent, said antifungal agent may be a fungicide or a fungistatic. The use of the strain of the invention, the supernatant of the invention, the compound of formula (I) of the invention or the composition of the invention in combination with other antibacterial agents is an interesting strategy in the prevention and / or treatment of bacterial infections, preferably of diseases of infectious origin caused by resistant or multi-resistant pathogenic bacteria, and in the inhibition of the formation of bacterial biofilms on any surface.
    [0104] Therefore, another aspect of the invention relates to the use of the supernatant of the invention, of the compound of formula (I) of the invention or of the composition of the invention for the manufacture of a medicament for the treatment and / or prevention of infections. bacterial. Alternatively, this aspect of the invention relates to the supernatant of the invention, the compound of formula (I) of the invention, or the composition of the invention for use in the treatment and / or prevention of bacterial infections.
    [0105] The bacterial infections to which the present invention refers are caused by one or more Gram positive or Gram negative bacteria, or both together. In a more preferred embodiment, the bacterial infections referred to in the present invention are caused by one or more bacteria from the selected genera of Corynebacterium, Staphylococcus, Streptococcus, Enterococcus, Clostridium Mycobacterium, Bacteroides, Haemophilus or Neisseria. In an even more preferred embodiment, the bacterial infections referred to in the present invention are caused by one or more bacteria selected from the list consisting of: Corynebacterium urealyticum, Staphylococcus aureus, Streptococcus pneumoniae, Streptococcus pyogenes, Enterococcus faecium, Enterococcus faecalis , Clostridium perfringens, Mycobacterium tuberculosis, Bacteroides fragilis, Haemophilus influenzae or Neisseria meningitidis.
    [0106] In another preferred embodiment, the bacteria referred to in the present invention are resistant bacteria, more preferably multi-resistant, to one to more antibiotics.
    [0107] Examples of antibiotic resistant bacteria include those strains resistant to aminoglycosides, carbapenems, cephalosporins, glycopeptides, lincosamides, lipopeptide, macrolides, monobactam, nitrofurans, oxazolidonones, penicillins, quinolones, sulfonamides, fusidic acid, pseudomonic acid, pseudomonic acid novobiocin and / or tetracyclines, among others. More preferably, the bacteria referred to in the present invention are bacteria resistant to at least one antibiotic selected from the list consisting of: amikacin, amoxicillin, ampicillin, capreomycin, ciprofloxacin, clavulanic acid, clindamycin, cotrimoxazole, ethambutol, erythromycin, phosphomycin, isoniazid, kanamycin, nitrofurantoin, quinolones, rifampin, streptomycin and / or tetracycline.
    [0108] In an even more preferred embodiment, the compound of formula (1) is desertomycin G and the bacteria is Gram-positive such as Mycobacterium tuberculosis, Corynebacterium urealyticum, Staphylococcus aureus, Streptococcus pneumoniae, Streptococcus pyogenes, Enterococcus faecium and Enterococcus faecalis perfringens.
    [0109] In another preferred embodiment, the compound of formula (1) is desertomycin G and the bacterium is Gram-negative such as Bacteroides fragilis, Haemophilus influenzae and / or Neisseria meningitidis.
    [0110]
    [0111]
    [0112] In another preferred embodiment, the medicine to which the present invention refers is for the treatment and / or prevention of bacterial infectious diseases or diseases caused by bacterial infections. More preferably, said diseases are selected from the list consisting of: tuberculosis, necrotic enteritis, gas gangrene, genitourinary tract infections (such as cystitis, urethritis, prostatitis, pyelonephritis, epididymitis), skin infections (such as impetigo, folliculitis, cellulitis , abscesses, wound infection), osteomyelitis, mastitis, pharyngotonsillitis, conjunctivitis, epiglottitis, otitis meningitis, meningococcemia, bacteremia, sepsis, endocarditis, respiratory tract infections (such as pneumonia or sinusitis), intra-abdominal infections (cholecystitis, peritonitis, visceral abscesses ), as well as nosocomial infections.
    [0113]
    [0114] The term "medicine" as used in this invention refers to any substance used for the prevention, relief, treatment or cure of infections or treatment of tumors in man, or any other animal, and plants. In the context of the present invention, this term refers to a preparation comprising the supernatant of the invention, the compound of formula (I) of the invention or the composition of the invention.
    [0115]
    [0116] The medicine to which the present invention refers can be for human or veterinary use. The "medicine for human use" is any substance or combination of substances that is presented as possessing properties for the treatment or prevention of diseases in humans or that can be used in humans or administered to humans in order to restore, correct or to modify the physiological functions exerting a pharmacological, immunological or metabolic action, or to establish a medical diagnosis. "Veterinary medicine" is any substance or combination of substances that is presented as having curative or preventive properties with respect to animal diseases or that can be administered to the animal in order to restore, correct or modify its physiological functions by exercising a pharmacological, immunological or metabolic action, or to establish a veterinary diagnosis, including, but not limited to, drug premixes. "Medicated premix" or "medicated food premix" means any veterinary medicinal product prepared in advance for the further manufacture of medicated food. "Medicated feed" is understood to mean any mixture of veterinary drug (s) and feed (s) prepared prior to marketing and intended to be administered to animals without transformation, due to the curative or preventive properties or other properties of the drug.
    [0117]
    [0118] The drugs of the invention can be used both alone and in combination with other drugs or compositions for the treatment or prevention of bacterial infections or for the treatment of tumor processes. Thus, the drugs of the present invention can be used together with other active ingredients or therapies as a combination therapy. The other active ingredients may be part of the same composition or may be provided by a different composition, being administered at the same time or at different times. The composition of the present invention, preferably pharmaceutical or cosmetic, may alternatively or additionally include another antitumor compound.
    [0119]
    [0120] The term "treatment", as understood in the present invention, refers to combating the effects caused as a consequence of the infection, disease or pathological condition of interest in a subject (preferably mammal, and more preferably a human) that includes:
    [0121] i. inhibit infection, disease or pathological condition, that is, stop its development;
    [0122] ii. alleviating the infection, disease or pathological condition, i.e. causing regression of the infection, disease or pathological condition or its symptoms;
    [0123]
    [0124] iii. stabilize infection, disease, or disease condition.
    [0125] The term "prevention", as understood in the present invention, consists in avoiding the appearance of infection or infectious disease or bacterial biofilm, that is, preventing the appearance of a biofilm on a surface or preventing infection from occurring or infectious disease in a subject (preferably mammal, and more preferably a human), particularly when said subject is predisposed to suffer an infection but has not yet been diagnosed as having it, as is the case, for example, in neonates, the elderly or immunosuppressed patients or recently undergone surgical intervention.
    [0126]
    [0127] The term "infection" is the clinical term used to describe the colonization of a host organism by microorganisms from other species. Clinically, the colonizing organism is detrimental to the normal functioning and survival of the host. As used herein, the term "bacterial infectious diseases" or "diseases of infectious origin caused by bacteria" refers to diseases preceded by a bacterial infection, including systemic infections (bacteremia and sepsis) and infections in any organ or tissue of the host organism . Organs or tissues include, but are not limited to, skeletal muscle, skin, soft or mucosal tissues, bloodstream, kidneys, or any other tissues of the urinary tract, respiratory tract, gastrointestinal tract, sexual organs, ear, eye, heart, lungs, or bone . These infections can be caused by Gram positive and / or Gram negative bacteria.
    [0128]
    [0129] In the present invention, said infections occur in a subject that can be, but is not limited to, an animal, in particular a mammal and more particularly a human, or a domestic animal, for example pig, mouse, rat, cat, gerbil, rabbit , dog, monkey, chimpanzee, etc., or a bird.
    [0130]
    [0131] Another aspect of the invention relates to the use of the strain of the invention, of the supernatant of the invention, of the compound of formula (I) of the invention or of the composition of the invention for the elimination and / or prevention and / or inhibition of the formation of bacterial biofilms, preferably on inert surfaces, that is to say ex vivo.
    [0132]
    [0133] In a preferred embodiment of this aspect of the invention, bacterial biofilms are caused by one or more bacteria from the selected genera of Clostridium, Corynebacterium, Enterococcus, Staphylococcus, Bacteroides, Escherichia, Haemophilus or Neisseria. In a more preferred embodiment, the bacterial biofilms are caused by bacteria selected from the list consisting of: Clostridium perfringens, Corynebacterium urealyticum, Enterococcus faecalis, Staphylococcus aureus, Bacteroides fragilis, Escherichia coli, Haemophilus influenzae or Neisseria meningitidis.
    [0134]
    [0135] "Biofilms" or biofilms are understood as the communities of microorganisms that grow embedded in a matrix of exopolysaccharides and attached to an inert surface or to tissue in vivo or ex vivo (in culture). They are communities of bacteria (single or multiple species) that adhere to a solid surface. Biofilms are a common cause of bacterial infections, both in humans and in other animals and plants. Examples of biofilms are those that form in the oral cavity, such as caries (dental plaque) or periodontal disease.
    [0136]
    [0137] The mechanisms by which the biofilm produces the symptoms of the disease are not yet fully established, but it has been suggested that the biofilm bacteria can produce exotoxins, groups of bacteria can be released into the bloodstream, become resistant to the phagocytic action of the cells of the immune system and, on the other hand, constitute a niche for the emergence of bacteria resistant to antibiotic treatments. This last aspect may be especially relevant since resistant bacteria originating from a biofilm could spread from patient to patient through the hands of healthcare personnel.
    [0138] On the other hand, the biological contamination of surfaces by the formation of biofilms is common, and the biofilm can develop on hydrophobic, hydrophilic, biotic or abiotic surfaces, and leads to the degradation of the material, contamination products, mechanical blocking and impedance of the transfer of heat in aquatic processes. Biofilms are also the leading cause of biological contamination in food, catheters, drains or implants, as well as in drinking water distribution systems and other pipelines, with biofilm control being especially important in fire fighting systems.
    [0139]
    [0140] The term "Gram positive" refers to bacteria that stain dark blue or violet in the Gram staining protocol and include, but are not limited to, Staphylococcus (including Staphylococcus aureus, S. epidermidis, S. haemolyticus, S. hominis, S. saprophytics), Streptococcus (including Streptococcus pneumoniae, S. pyogenes, S. agalactiae, S. avium, S. bovis, S. lactis, S. sanguis and Streptococcus of group C, Streptococcus of group G and Streptococcus viridans), Enterococcus (including Enterococcus faecalis and E. faecium), Clostridium, Corynebacterium and Mycobacterium tuberculosis. Common S. aureus strains are resistant to penicillin. The appearance of strains of this species resistant to methicillin and vancomycin represents a serious health problem.
    [0141]
    [0142] The term "Gram negative" refers to bacteria that do not retain violet staining in the Gram staining protocol and include, but are not limited to, Enterobacteriaceae, including E. coli, Klebsiella spp., Haemophilus, Bacteroides, Enterobacter spp., Citrobacter spp., Serratia spp., Proteus spp., Providencia spp., Salmonella spp., Shigella spp., Pseudomonas (including P. aeruginosa) and species such as Moraxella spp. (Including M. catarrhalis) and Neisseria spp.
    [0143] The use of the strain of the invention, of the supernatant of the invention, of the compound of formula (1) of the invention or of the composition of the invention can be used both in the treatment of cancer and in the prevention and / or treatment of bacterial infections, preferably diseases of infectious origin caused by resistant or multi-resistant pathogenic bacteria, and in the inhibition of the formation of bacterial biofilms on any surface.
    [0144]
    [0145] Therefore, another aspect of the invention relates to the use of the supernatant of the invention, of the compound of formula (I) of the invention or of the composition, preferably pharmaceutical, of the invention, for the manufacture of a medicament. Alternatively, this aspect of the invention relates to the supernatant of the invention, the compound of formula (I) of the invention or the composition of the invention for use as a medicine.
    [0146]
    [0147] Another aspect of the invention relates to the use of the supernatant of the invention, of the compound of formula (I) of the invention or of the composition of the invention for use in the treatment and / or prevention of bacterial infections. Alternatively, this aspect of the invention relates to the supernatant of the invention, the compound of formula (1) of the invention or the composition of the invention for the manufacture of a medicine for the treatment of cancer.
    [0148]
    [0149] The compound of formula (1) of the invention is an inhibitor of tumor growth and is therefore useful in the treatment of cancer.
    [0150]
    [0151] Thus, pharmaceutical compositions comprising an effective amount of a compound of formula I or a pharmaceutically acceptable salt or solvate thereof together with a pharmaceutically acceptable excipient are included in the present description.
    [0152] Also included in the present description is the use of a compound of formula I or a pharmaceutically acceptable salt or solvate thereof in the manufacture of a medicament.
    [0153]
    [0154]
    [0155] Also included in the present disclosure is the use of a compound of formula I or a pharmaceutically acceptable salt or solvate thereof to inhibit the growth of a tumor.
    [0156] As used here, "inhibit" means decrease, slow down, or stop. Thus, a compound of this invention can decrease, slow down, or stop the growth of a tumor cell. As used herein, "growth" means increase in size, or proliferation, or both. Thus, a compound of this invention can inhibit the increase in size of a tumor cell and / or can prevent the tumor cell from dividing and increasing the number of tumor cells. A "tumor cell" is a cell that constitutes a neoplasm (new growth), which can be cancerous (malignant) or non-cancerous (benign). A cancer tumor cell can invade normal tissues around it and blood / lymphatic vessels and metastasize to tissues far from the original tumor. Conversely, a non-cancerous tumor cell can grow and compress adjacent normal tissues but cannot invade normal tissues and blood / lymphatic vessels, nor can it metastasize to tissues far from the original tumor.
    [0157] Also included in the present description is the use of a compound of formula I or a pharmaceutically acceptable salt or solvate thereof for treating cancer.
    [0158] Also included in the present description is the use of a compound of formula I or a pharmaceutically acceptable salt or solvate thereof in the manufacture of a medicament with antitumor activity.
    [0159] Also included in the present description is the use of a compound of formula I or a pharmaceutically acceptable salt or solvate thereof in the manufacture of a medicament for the treatment of cancer.
    [0160] Also included in the present description is a method of treating a subject, including a human, diagnosed with cancer, which consists of treating said subject with a therapeutically effective amount of a compound of formula I or a pharmaceutically acceptable salt or solvate.
    [0161] In another preferred embodiment, the medicament to which the present invention refers is for the treatment of cancer by inhibiting the growth of the cells that make up the tumor, thereby preventing invasion of normal tissues and blood / lymphatic vessels by tumor cells and therefore prevents metastasis. Examples of cancers that can be treated include breast and colon, but are not limited to pancreas, ovary, prostate, testis, melanoma, kidney, central nervous system, and leukemia (etc.). The term "acceptable pharmaceutical composition" consists of a biologically suitable material, that is, the material can be administered to the subject without causing substantially harmful biological effects.
    [0162] Throughout the description and claims, the word "comprises" and its variants are not intended to exclude other technical characteristics, additives, components or steps. For those skilled in the art, other objects, advantages and characteristics of the invention will emerge in part from the description and in part from the practice of the invention. The following examples and figures are provided by way of illustration, and are not intended to be limiting of the present invention.
    [0163] Brief description of the drawings
    [0164] To complement the description that is being made and in order to help a better understanding of the characteristics of the invention, it is included as an integral part of said description, a set of drawings in which, by way of illustration and not limitation, the following has been represented:
    [0165] FIG. 1. Phylogenetic tree obtained by the "neighbor-joining" method obtained by matrix distance analysis of the 16S rRNA gene sequences, showing the position of Streptomyces althioticus MSM3 and its closest phylogenetic relatives. The node numbers are bootstrap values (1000 samples; only values> 50% are presented). Asterisks indicate that the corresponding nodes were also obtained in the tree obtained by the maximum-likelihood method. The bar indicates 0.05% sequence divergence.
    [0166] FIG. 2. Proliferation assay of cancer cell lines A549, DLD-1, MCF-7 and healthy mammary fibroblasts in the presence or absence of different concentrations of Desertomycin G. Cell proliferation rates were determined on five consecutive days using an automatic reader of microtiter plates. Significant differences were determined with Student's t-test and p values less than 0.05 were considered statistically significant (p <0.05, * p <0.01, "p <0.005, ***).
    [0167] PREFERRED EMBODIMENT OF THE INVENTION
    [0168] The invention will now be illustrated by means of tests carried out by the inventors, which demonstrate the effectiveness of the strain of the invention in the production of a new natural product of the desertomycin family (designated in the present invention as desertomycin G) , as well as antimicrobial activity against both Gram positive and Gram negative pathogenic bacteria and cytotoxic activity against human tumor lines. Example 1. Experimental section
    [0169] General experimental procedures
    [0170] Analysis and separations by semi-preparative HPLC were carried out using an Alliance chromatographic system with a SunFire C18 column (10 pm, 10 x 250 mm, Waters). For UPLC analyzes, an UPLC Acquity equipped with a BEH C18 column (1.7 pm, 2.1 x 100 mm, Waters) was used. Optical rotation was determined with a JASCO P-2000 polarimeter. IR spectra were measured with a JASCO FT / IR-4100 spectrometer equipped with a PIKE MIRacle ™ ATR accessory (single reflection). NMR spectra were recorded on a Bruker Avance III spectrometer (500 and 125 MHz for 1H and 13C NMR, respectively) equipped with a 1.7 mm TCI MicroCryoProbe ™ probe, using the residual solvent signal as internal reference (5 hr. 7.27 and 5e 77.0 ppm for CDCI 3 ). HRESIMS spectra were acquired using a Bruker maXis QTOF mass spectrometer.
    [0171] Microorganisms and fermentation conditions
    [0172] In the course of our search for new antibiotic-producing Actinobacteria, a marine strain of Streptomyces MSM3 (CECT 9651) was isolated from the Ulva sp. Macroalgae, collected in the Cantabrian Sea in Pedreña, Cantabria (coordinates 43 ° 26 37 "N, 3 ° 46 5 "W). 30 Erlenmeyer flasks (250 ml), each containing 50 ml of R5A medium (Braña et al., 2015, Microb Ecol 69: 512-24) were inoculated with spores of this strain and incubated on an orbital shaker at 28 ° c and 250 rpm for 6 days.
    [0173] Phyloanetic analysis (taxonomy) of the producer microoraanism
    [0174] The MSM3 strain (CECT 9651) was subjected to phylogenetic analysis based on the sequence analysis of the 16S rRNA. Phylogenetic analysis was performed using MEGA version 6.0 after
    [0175]
    [0176]
    [0177] of a multiple alignment of the data by CLUSTALO. Distances (distance options according to the Kimura two-parameter model) and alignment with the neighborjoining method were determined using bootstrap values based on 1000 replications.
    [0178]
    [0179] Antimicrobial activity of desertomycin G against clinical pathogens
    [0180]
    [0181] The antimicrobial activity of desertomycin G and the minimum inhibitory concentration (MIC) against a group of human pathogens were determined (Table 1).
    [0182]
    [0183] Clinical pathogens Isolated Hospital Year Antibiotic resistance
    [0184] Gram-positive
    [0185] Mycobacterium tuberculosis H37Rv ATCC 27294
    [0186] Mycobacterium tuberculosis MDR-1 14,595 SNRL-Spain 2013 Multidrug resistant Mycobacterium tuberculosis MDR-2 14,615 SNRL-Spain 2013 Multidrug resistant Clostridium perfringens 103281 HUCA 2013 -Corynebacterium urealyticum 1492 Cabueñes 2014 MuItirresistantb Enterococcus faecalis 10544 Cabuec
    [0187] Enterococcus faecalis ATCC 29212
    [0188] Enterococcus faecium 10701 Cabueñes 2015 Amp, quin, ery Streptococcus pneumoniae 64412 HUCA 2013 Ery Streptococcus pyogenes 81293 HUCA 2013 -
    [0189] Staphylococcus aureus 11497 Cabueñes 2015 Susceptible methicillin Staphylococcus aureus ATCC 43300
    [0190]
    [0191] Staphylococcus aureus ATCC 25923
    [0192]
    [0193] Gram-negative
    [0194]
    [0195] Bacteroides fragilis 61592 HUCA 2013 Amo, tet
    [0196]
    [0197] R es is te nc ia sa A p la genoc y A is la do H osp ita l A n y an tib ió tic os
    [0198] B a c te ro id e s fra g ilis A T C C 25285 -H a e m o p h ilu s in fiu en zae 10996 C a b u e ñ e s 2015 A m p, co t, qu in
    [0199] H a e m o p h ilu s in flu en za A A C C 49 247 -N e isse ria m en in g itid is 71327 H U C A 2013 C lin
    [0200]
    [0201] Table 1. Description of clinical pathogens. a Inh, rif, emb.b lnh, rif, emb, str, amk, kan, cap. C Amp, amo / clav, ery, cot, cip, fos, nitro. Amk: amikacin; master: amoxicillin; amp: ampicillin; cap: capreomycin; cip: ciprofloxacin; clav: clavulanic acid; clin: clindamycin; cot: cotrimoxazole; emb: ethambutol; ery: erythrornicin; fos: phosphomycin; inh: isoniazid; kan: kanamycin; nitro: nitrofurantoin; quin: quinolones; rif: rifampicin; str: streptomycin; tet: tetracycline.
    [0202]
    [0203] Some of them have been isolated and identified in clinical microbiology laboratories from samples obtained from patients with clinical infections. The Mueller-Hinton medium (Biomedics) was used in the bioassays against Escherichia coli, Staphylococcus aureus, Enterococcus faecalis, Enterococcus faecium, Haemophilus influenzae, being supplemented with
    [0204]
    [0205]
    [0206] according to the CLSI conditions (CLSI, Clinical and Laboratory Standards Institute, document M100-S24 2014) for Streptococcus pneumoniae, Streptococcus pyogenes and Neisseria meningitidis. Tryptic soybean with ram blood w / 5% (DIFCO) was used for Cotynebacterium urealyticum. Brucella Broth (SIGMA) supplemented with hemin (5 qg / ml), vitamin K1 (1 qg / ml) and lysed horse blood (5% v / v) was used for Bacteroides fragilis and Clostridium perfringens.
    [0207]
    [0208] For most Gram-positive and Gram-negative bacteria, antimicrobial assays were performed according to CLSI protocol standards. Mycobacterium tuberculosis sensitivity tests were performed in Middlebrook 7H10 agar medium supplemented with 10% OADC and 0.5% glycerol according to the agar ratio method for slow growing mycobacteria (CLSI document M24-A2, 2011).
    [0209]
    [0210] Example 2. Results
    [0211]
    [0212] Taxonomy of strain MSM3
    [0213]
    [0214] The 16 S rDNA of the producer strain MSM3 (CECT 9651) was amplified by polymerase chain reaction (PCR) and sequenced as previously described (Sarmiento-Vizcaíno et al., 2016, Microb Ecol. DOI 10.1007 / s00248- 016-0845-2). Sequence analysis demonstrated 99.9% identity with Streptomyces althioticus (AY999808) therefore this strain was designated as Streptomyces althioticus MSM3 (EMBL sequence number LT627193). Another strain of this species Streptomyces althioticus (KCTC 9752), producer of an unidentified anti-tuberculosis compound, was isolated from the soil of the Thar desert, Rajasthan (Radhakrishnan et al., 2013, Bioinformation 9: 18-22. Doi: 10.6026 / 97320630009018 ). The phylogenetic tree generated by the "neighbor-joining" method, based on the 16S rRNA gene sequence, clearly revealed the evolutionary relationship of the MSM3 strain with a group of known Streptomyces species (Fig. 1).
    [0215]
    [0216] Isolation and purification of desertomycin G
    [0217]
    [0218] Cultures were centrifuged, pellets were discarded, and supernatants were filtered and applied to a solid phase extraction cartridge (Sep-Pak Vac C18, 10g, Waters). The retained material was eluted with a mixture of methanol and 0.05% trifluoroacetic acid (TFA) in water, using a linear gradient from 0 to 100% methanol in 60 min, at 10 ml / min. Fractions were collected every 5 minutes and analyzed by UPLC using described chromatographic conditions (Braña et al., 2014, Arch Microbiol .; 196: 345-355). In fractions taken between 35 and 45 min, a peak corresponding to the desired compound was observed. These fractions were pooled, partially vacuum dried and applied to a solid phase extraction cartridge (C18 Sep-Pak Vac, 2g, Waters). The cartridge was washed with water, the retained compounds were eluted with methane 'and dried under vacuum. The residue was subsequently redissolved in a small volume of methanol and DMSO (1: 1). Purification was performed in two steps using a SunFire C18 column (10mm, 10 x 250mm, Waters). In the first step the extract was chromatographed with a mixture of acetonitrile and 0.1% TFA in water (32:68) under isocratic conditions and a flow of 7 ml / min. In the second step, the mobile phase was a mixture of methanol and 0.1% TFA in water (67:33), at 6 ml / min. After each step, the solution containing the collected peak was partially evaporated in vacuo to reduce the organic solvent concentration and then applied to a solid phase extraction cartridge (Sep-Pak C18, 500 mg, Waters). The cartridge was washed with water and the retained compound was eluted with methanol. The purified compound was finally redissolved in a mixture of tert-butanol and water (1: 1) and lyophilized, resulting in 20.4 mg of product in its pure state.
    [0219]
    [0220] Desertomycin G: white solid; [a] 20 d +15.6 ° (c 0.35, MeOH); IR (ATR) vmax 3367, 2970, 2927, 1683, 1637, 1455, 1433, 1387, 1255, 1232, 1203, 1136, 1105, 1070, 1053, 971 cmr1; for the 1H and 13C NMR data see Table 1; HRESIMS m / z 12047609 [M + H] + (calcd. For C 62 H 110 NO 21 +, 1204.7565), 593.8776 [M-H20 + 2H] 2+ (calcd. For C 62 H 109 NO 20 2+,, 5938774).
    [0221]
    [0222] Structural elucidation
    [0223]
    [0224] Desertomycin G has the molecular formula C 62 H 109 NO 21 according to the protonated ion am / z 1204.7906 in its ESI-TOF spectrum (calcd for C 62 H 110 NO 21 + 1004.7565, A 3.6 ppm). Analysis of 1H, 13C and HSQC spectra revealed the presence in the molecule of 10 olefinic protons, 18 oxygenated methins, a doubly oxygenated methine, probably belonging to a sugar residue, an oxygenated methylene, 8 aliphatic methins, 8 aliphatic methylenes and aliphatic methyl groups, suggesting a polyketonic nature for the compound.
    [0225]
    [0226] The correlations observed in the COZY spectrum allowed establishing the sequence of C-3 to C-19 and C-21 to C-46 and also allows the union of the methyl groups C-48, C-49, C-50, C- 51, C-53, C-54, C-55 and C-56 C-6, C-8, C-14, C-18, C-24, C-30, C-32 and C-42, respectively . On the other hand, the HMBC correlations between H3-47 C-1, C-2 C-3 and H3-52 to C-19, C-20 and C-20 allowed completing the linear sequence from C-1 to C- 46. An additional HMBC correlation between H-41 and C-1 and the unshielded chemical shift of H-41 (5.11 ppm) were also indicative of the existence of a lactone ring between C-41 and C-1. Finally, the only nitrogen atom present in the molecule is associated with the presence of a primary amino group at C-46, evidenced by the corresponding chemical shift of 1H and 13C NMR at this terminal position (5h 2.93 ppm and 5c 40 8 ppm). A bibliographic search has established that the planar structure of this macrocyclic residue of the molecule was very similar to that found in desertomycin A, the two differences between both molecules being the presence of an additional double bond in A4 and an additional methyl group (C -53), located at C-24 in the structure of desertomycin G. The remaining not assigned signals corresponded to a doubly oxygenated methine group, four oxygenated methines and one oxygenated methylene (C-1 'to - C-6') . These signals are in agreement with the presence in the molecule of a hexopyranose that was identified as a.-D-mannopyranose based on chemical shifts similar to those measured in the structure of desertomycin A (Bax et al. 1986, J Am Chem Soc 108: 8056-8063). Furthermore, HMBC correlations from H-1 'to C-22 and from H-22 to C-1' confirmed the binding of this sugar molecule to C-22 carbon.
    [0227]
    [0228]
    [0229] Table 2. 1 H and 13 C NMR spectra for desertomycin G (CD 3 OD, 500 MHz)
    [0230]
    [0231]
    [0232]
    [0233]
    [0234]
    [0235]
    [0236] Antimicrobial activity of desertomycin G
    [0237]
    [0238] The antimicrobial activity of the compound was tested against a group of human pathogens (Table 1). Some of them have been isolated and identified in clinical microbiology laboratories from samples obtained from patients with clinical infections.
    [0239]
    [0240] Table 3. Minimum inhibitory concentrations (MIC) against a panel of clinical pathogens.
    [0241]
    [0242] DesertomycinG
    [0243] MIC (pq / ml)
    [0244]
    [0245] Gram-positive
    [0246] Corynebacterium urealyticum 1492 <0.25
    [0247] Staphylococcus aureus 11497 4
    [0248] Staphylococcus aureus ATCC 25923 4
    [0249] Staphylococcus aureus ATCC 43300 4
    [0250] Streptococcus pneumoniae 64412 4
    [0251] Streptococcus pyogenes 81293 4
    [0252] Enterococcus faecium 10701 4
    [0253] Enterococcus faecalis 10544 8
    [0254] Enterococcus faecalis ATCC 51299 8
    [0255] Enterococcus faecalis ATCC 29212 8
    [0256] Clostridium perfringens 103 281 16
    [0257] Mycobacterium tuberculosis H37Rv 16
    [0258] Mycobacterium tuberculosis MDR-1 16
    [0259] Mycobacterium tuberculosis MDR-2 16
    [0260]
    [0261] Gram-negative
    [0262] Bacteroides fragilis 61592 32
    [0263] Bacteroides fragilis ATCC 25285 32
    [0264] Haemophilus influenzae 10996> 64
    [0265] Haemophilus influenzae ATCC 49247 64
    [0266] Neisseria meningitidis 71327 64
    [0267]
    [0268]
    [0269] Table 3 shows the minimum inhibitory concentrations (MIC). The compound shows strong activities against Gram-positive pathogenic bacteria and Mycobacterium tuberculosis, Corynebacterium urealyticum, Staphylococcus aureus, Streptococcus pneumoniae, Streptococcus pyogenes, Enterococcus faecium, Enterococcus faecalis, Clostridium perfringens; and moderate against Gram-negative bacteria Bacteroides fragilis, Haemophilus influenzae, Neisseria meningitidis.
    [0270]
    [0271] Cytotoxic activity of desertomycin G
    [0272]
    [0273] Cytotoxic activity was observed for the compound against human cell lines A549 for lung carcinoma, DLD-1 for colon carcinoma and MCF-7 for breast adenocarcinoma, as well as in healthy mammary fibroblasts. On the third day, the results indicate that the DLD-1 and MCF-7 cell lines decrease their viability by about 50% compared to the control. When using 2.5 and 5 pm desertomycin G, the A549 line is more resistant and healthy mammary fibroblasts remain unaffected at these concentrations (Fig. 2).
    [0274]
    [0275] In conclusion, a new natural product, desertomycin G, was obtained from Streptomyces althioticus MSM3, isolated from the intertidal algae Ulva sp. collection in the Cantabrian Sea (Pedreña, Cantabria). Desertomycin G is a novel natural product and is also the first member of the family to show strong antibiotic activity against Mycobacterium tuberculosis and other Gram-positive pathogens. Due to its antibiotic activity against resistant clinical pathogens and given the medical needs of new antibiotics against these microorganisms, desertomycin G deserves to be considered as a candidate for antibiotic chemotherapy, especially against resistant Mycobacterium tuberculosis but also against the rest of the indicated pathogens. Furthermore, cytotoxicity against tumor cell lines but not against mammary fibroblasts highlights the use of desertomycin G as a potential antitumor agent. These results are an example of the relevance of natural marine products as candidates in antibacterial and antitumor chemotherapy.
    权利要求:
    Claims (20)
    [1]
    1. Bacterial strain of Streptomyces althioticus MSM3 deposited in the Spanish Type Culture Collection under accession number CECT 9651,
    [2]
    2. A culture supernatant of the strain according to claim 1.
    [3]
    3. Use of the strain according to claim 1 for the production of the compound of formula (I) any of its salts or tautomers:

    [4]
    4. Process for obtaining a compound of general formula (I), or any of its salts or tautomers, described in claim 3, comprising:
    to. cultivating the strain according to claim 1 in a culture medium under fermentation conditions, and
    b. purify the compound of formula (I) produced by the cultivated strain of step (a).
    [5]
    5. Compound of general formula (I):

    [6]
    6. A compound according to claim 5 wherein the group R 1 is selected from hydrogen, substituted or unsubstituted methyl, substituted or unsubstituted ethyl, substituted or unsubstituted n-propyl, substituted or unsubstituted iso-propyl , substituted or unsubstituted n-butyl, substituted or unsubstituted tert-butyl, substituted or unsubstituted vinyl, substituted or unsubstituted allyl, substituted or unsubstituted ethynyl, substituted or unsubstituted acetyl, substituted or unsubstituted benzyl, and substituted or not replaced.
    [7]
    7. A compound according to claim 6 which group R 2 is selected from hydrogen, acetyl, benzyl and benzoyl.
    [8]
    8. A compound according to claim 6 which group R 2 is hydrogen.
    [9]
    9. A compound according to any of claims 5-8 wherein the groups R 2 and R 3 are independently selected from hydrogen, COR a , COOR a and substituted or unsubstituted C 1 -C 6 alkyl, where Ra is selected between hydrogen and substituted or unsubstituted C 1 -C 6 alkyl.
    [10]
    10. A compound according to claim 9 wherein Ra is selected from methyl, ethyl, n-propyl, iso-propyl and butyl, including n-butyl, tert-butyl, sec-butyl and iso-butyl.
    [11]
    11. A compound according to claim 9 wherein the groups R3 and R4 are hydrogen.
    [12]
    12. The compound of claim 5 with the following formula (II):

    [13]
    13. Composition comprising the supernatant according to claim 2 or the compound according to claims 5 to 12.
    [14]
    14. Composition according to claim 13, which further comprises another antibacterial and / or antitumor agent.
    [15]
    15. Composition according to any of claims 14 or 15, wherein the composition is a pharmaceutical or cosmetic composition and further comprises a pharmaceutically or cosmetically acceptable carrier or excipient.
    [16]
    16. Use of the supernatant according to claim 2, of the compound according to claims 5 to 13 or of the composition according to any of claims 14 or 15, for the manufacture of a medicament.
    [17]
    17. Use of the supernatant according to claim 2, of the compound according to claims 5 to 13 or of the composition according to any of claims 14 or 15, for the manufacture of a medicament for the treatment and / or prevention of bacterial infections and / or for the treatment of tumors.
    [18]
    18. Use of claim 17, where the bacterial infections are caused by Gram-positive bacteria such as Mycobacterium tuberculosis, Staphylococcus aureus corynebacterium urealyticum, Streptococcus pneumoniae, Streptococcus pyogenes Enterococcus faecium, Enterococcus faecalis and Clostridium perfringens.
    [19]
    19. Use of claim 17, where the bacterial infections are caused by Gram-negative bacteria such as Bacteroides fragilis, Haemophilus influenzae, Neisseria meningitidis.
    [20]
    20. Use of claim 17, wherein the tumors are breast or colon.
    two
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    公开号 | 申请日 | 公开日 | 申请人 | 专利标题
    US3642984A|1964-04-28|1972-02-15|Upjohn Co|Antibiotic garlandosus and process for preparing the same|
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