奥地利专利AT510585A4 COMPOSITION COMPRISING A PEPTIDE AND AN INHIBITOR OF VIRAL NEURAMINIDASE

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专利摘要:
Disclosed is a composition comprising a peptide consisting of 7-17 contiguous amino acids and comprising the hexamer TX1EX2X3E, wherein X1, X2 and X3 may be any natural or non-natural amino acid, which peptide has no TNF receptor binding activity and cyclizes and an inhibitor of viral neuraminidase, in particular for use in the prevention and treatment of influenza.
公开号:AT510585A4
申请号:T1908/2010
申请日:2010-11-18
公开日:2012-05-15
发明作者:Bernhard Dr Fischer；Rudolf Dr Lucas；Hendrik Fischer
申请人:Apeptico Forschung & Entwicklung Gmbh；
IPC主号:

专利说明:

1
The present invention relates! pharmaceutical compositions for the treatment of influenza.
In humans, influenza is a severe disease of the arterial pathways and the whole organism caused by influenza viruses. The influenza viruses belong to the family of orthomyxoviruses, which are characterized by a segmented RNA genome in negative strand orientation. The human-relevant representatives are the influenza A and B viruses, whereby the subtype Δ in particular is known to be the causative agent of highly febrile respiratory diseases. In addition to veterinary importance, all influenza viruses have a zoonitic potential, i. a transfer from the chicken or pig to humans is possible.
The influenza periodically occurs as a pandemic, which usually originates in Southeast Asia and China and spreads from there worldwide. Influenza virus pandemics are associated with a high number of deaths not only in the elderly, but also in adolescents. According to the WHO, annual seasonal influenza causes around 3 to 5 million cases of severe disease worldwide, with death rates ranging from 250,000 to 500,000. The most common cause of death is influenza pneumonia with resulting. Lung failure, but also cardiovascular damage such as myocarditis (cardiac muscleIenizünduno) ocer pericarditis (pericarditis) occur. In addition, an inflammation of the brain or meninges (meningoencephalitis) or the damage of other organ systems (especially kidney) come as a more frequent cause of death in question.
The incubation period is usually 4-5 days, but shorter. The disease begins with the sudden onset of headache, chills, chills and coughing. High fever at 41 ° C, muscle pain, loss of appetite and general feeling of weakness. This phase lasts for about 3 days, after which the fever goes back and usually falls to normal values from the sixth day, the virus is eliminated from a body. The cough may last several weeks.
Severe, life-threatening influenza occurs when your primary viral interstitial infection (often haemorrhagic) develops after the symptoms described. It occurs in approximately 25% of healthy or undamaged persons, along with weakened individuals, and can last up to 2 weeks. Such a lung infection can be 2 -
be detected by the increase in lung weight.
Pneumonia may also be secondary to bacterial over-infection (including Streptococcus pneumoniae, Staphylococcus aureus and Haemophilus influenzae). Beneficial factors for these complications include, but are not limited to, other lung diseases {e.g. Asthma), inunsions, old age (infants and the elderly), diabetes, lung injuries, smoking. People with these complications are therefore the primary target group for vaccination.
Influenza viruses enter the organism through droplet infection and, by binding the HA protein, they infect terminal neuraminic acid residues on the epithelial cells of the oral, nasal and pharyngeal mucosa. From here they spread to the lower respiratory tract. Cell destruction can be observed in the ciliated epithelia and the mucous-producing skin layers of all areas of the respiratory tract. If primary, interstitial pneumonia develops, the virus is transmitted to the cells of the lung parenchyma. One finds strong swelling of the alveolar walls, whose epithelium is often completely removed by the cell destruction. Such swelling of lung tissue can be detected by the increase in lung weight.
There are both prophylactic and therapeutic treatments. For example, vaccines against influenza A and B infections are available. It is oabei urr. killed viruses grown in chicken eggs and / or cell culture. The vaccine has reached its full effectiveness about two weeks after the vaccination. Because of the high. Variability of in-vivo viruses, however, the vaccines must be adjusted annually to the currently circulating virus subtypes or subtype variants. Furthermore, inhibitors of the viral meuraminidase (Zana-ruvir, Ose ', tamivi r) are used, which prevent the I.os solution of newly replicated virus particles from the host cell. They are preferably used shortly after successful and successful infection with influenza virus to control the spread of virus in the early stages of infection.
The inhibition of viral neuraninidase, however, only affects the multiplication of viruses, but they can not inactivate already existing body viruses. Keuraminidase Exchangers can only reduce the duration of the illness slightly (on average by one day in adults).
It is the object of the present invention to significantly increase the therapeutic effect of such virus proliferation inhibitors. The present invention is intended to provide improved pharmaceutical compositions for the treatment of infections with influenza viruses.
Accordingly, the present invention relates to a composition comprising a peptide consisting of 7-17 contiguous amino acids and comprising the hexamer TX1EX2X3E, wherein Xi, X2 and X3 may be any natural or non-natural amino acid, the peptide having no TNF receptor binding activity and is cyclized, and an inhibitor of viral neuraminidase, in particular for use in preventing and treating the
Influenza.
It has only been found according to the invention that the effect of neuraminidase inhibitors can be surprisingly improved if a peptide as defined above is used in combination with a neuraminidase inhibitor for the treatment of influenza infections. A prophylactic use of the composition according to the invention is thus indicated. The present invention has proven particularly efficient in the treatment of pneumonia caused by influenza viruses.
The peptides to be used according to the invention are known, for example, from European Patent EP 1 264 539 31, and have been used in the prior art for the treatment of fluid collections (pulmonary edema) and in particular for the RcabsorpLion of these fluid collections. However, surprisingly, these peptides also lend themselves to counteracting the reverse flow of liquid from the capillary endothelium. in the
Therefore, they can also be used to prevent and treat the hyperpermeability of epileiols and endotheiceles: (WO 20) 3/099106 A).
These - known per se - peptides, which he together with. Neuromir.idasehernmer are used, have no TNF receptor binding activity. (Hribar et al., Eur. J. Immunol., 1999; Elia et al., AJRCCM 2003; see also Example Example 4 below) and are cyclized. Preferred variants of these peptides consist of 7-17 contiguous amino acids and contain the heparin TPEGAE (SEQ ID NO: 4).
In a particularly preferred embodiment of the present invention, the composition according to the invention comprises a cyclized peptide consisting of a sequence of consecutive amino acids selected from the group consisting of - QRETPEGAEAKPWY (SEQ ID NO: 5) - PKDTPEGAELKPWY (SEQ ID NO: 6) - CGQRETPEGAEAKPWYC ( SEQ ID NO: 1) and CGPKDTPEGAELKPWYC (SEQ ID NO: 7) and at least 7 amino acids thereof which have fragments of the hexamer TPEGAE.
Preferably, the peptide in the composition comprises the amino acid sequence CGQRETPEGAEAKPWYC (SEQ ID NO: 1) and is cyclized via the C residues. This particularly preferred peptide thus has the following amino acid sequence (SEQ ID No. 1) (NH 2) Cys-Gly-Gln-Arg-Glu-Thr-Pro-Glu-Gly-Ala-Glu-Ala-Lys-Pro-Trp-Tyr Cys (COOH). This peptide is also called " AP301 " designated .
The cyclization of the peptides according to the invention can thereby /. b. either by direct cyclization across a disulfide bridge between the two C-residues at the N and C terminus, or by coupling the peptide to a support via both cysteines. In this case, the cysteine residues are preferably provided in the peptides according to the invention at the beginning and at the end of the molecule. Other functional groups that achieve cyclization of the peptide can also be used, e.g. in which an acid group leads to an amide or ester ring closure with an amine or alcohol (in this case, for example, the amino acids aspartic acid and glutamic acid can be cyclized with serine, threonine, tyrosine, asparagine, glutamine or hysin, preferably intramolecularly). The cyclization of the peptide is preferably by a disulfic bridge between the C residues of the peptide (if present).
Further preferred peptides according to the invention are therefore, for example, CGQKETPEGAEAKPWYC (SEQ ID No. 8), CGQ RET P EGAEAR PWYC (SEQ ID No. 9), CGQRETPEGAEAKPC (SEQ ID No. 10),
CQRETPEGAEAKPWYC (SEQ ID NO: 11) or CGQRETPEGAEAKFWYC (SEQ ID 5 * * φ φ φ φ φ * φφφ • φ φ φ φ
No. 12).
However, the cyclization can also be effected by binding of the peptide to carrier substances. As such Zyklisierungs carriers are all common pharmaceutically acceptable substances in question, which are capable, for. covalent binding with the SH groups of cysteines (or with other naturally occurring or artificially introduced chemically reactive groups of the peptide), with common carrier proteins such as Keyhole lirr.pet hemocyanin (KLH), tetanus toxin, etc., are particularly suitable , Also, adjacent bifunctional residues may be provided on the support (e.g., acid group adjacent to amine or alcohol group). In this context, it is important that with "cyclization " both the intramolecular ring closure and the incorporation of a carrier (from which the bound peptide protrudes (by linking the N and C termini of the peptide to the carrier)), the cyclized peptide thus showing the cyclic spatial structure and correspondingly is stabilized.
Inhibitors of viral neuraminidase are known and have already been proven in the prevention and treatment of influenza. Such viral neuruminidase inhibitors (such as zanamivir, üseltamivir, laninamivir or Perami.vir) prevent the release of newly replicated viral particles from the host cell. Especially shortly after successful and proven infection with influenza viruses, the virus spread in the early phase of the infection can be contained.
However, these neuraminidase inhibitors intervene only in the multiplication of viruses, but they can not inactivate viruses already present in the body, so that often only the duration of the disease can be shortened slightly (in adults by one day). Above all, however, the symptoms of influenza, in particular pneumonia, which usually leads to particular complications, can generally not be effectively combated or prevented.
Neuraminidase is an essential enzyme for the reproduction of the influenza virus and it is called "molecular scissors". described which is responsible for the release of the virus.
Neuraminidase inhibitors include sialic acid analogs which are a new class of the second generation antiviral agents. 6 - 4 * * * * II ♦ · * · »» * ·· »*» »·» · ** »« ··· ·· 4 · show efficacy against influenza A and B viruses. Neuraminidase inhibitors according to the present invention may be all compounds heretofore proposed, e.g. in US 2008/0063722 A1 summarized (also for preferred pharmaceutical formulations of such substances). These substances may inhibit at least one enzymatic activity of the neuraminidase protein of a virulent strain of a type A or type B influenza virion. Such substances can be used both for the prophylaxis and for the treatment of influenza; however, in combination with the above-defined peptides according to the present invention, this effect is significantly improved.
Examples of neuraminidase inhibitors which can be used in the present composition are e.g. in U.S. Patents 5,453,533, 5,763,483, 5,952,375, 5,958,973, 5,512,596, 5,886,213, 5,602,277, 6,410,594, 5,360,817, 5,866,601, 6,340,702, 6,451 766, 6,455,571, 6,593,314, 6,509,359, 6,518,305 and 6,831,096. Substances already used in humans (or at least in clinical trials) and thus particularly preferred are, for example, CS-8958 (RI 18958; US 2008/0063722 A1), zanamivir (GG167, RELENZA'1 '), peramivir (RWJ-270201, BCX-1812), Osollamivir Phosphate (Ro64-0796, GS4104), Oseltamivir Carboxylate (Ro64-0802, GS4071) or Oseltamivir (GS4104, ΤΛΜΙΓΓ.ΙΓ '). Of course, according to the invention, the neuraminic acid in all effective chemical. Forms, that is, as salts, racemic, optically pure and / or saltless forms (also in the form of, for example, enantiomers or diastereomers).
Preferably, the inhibitor of the viral neuraminidase is Zunamivir or oseltamivir; These substances are preferred because they are already used with particular success for the treatment of humans.
Preferably, the composition of the invention comprises a pharmaceutically acceptable carrier and is prepared as a pharmaceutical composition suitable for human administration. not interfering with each other negatively)
The term " a pharmaceutical composition " refers to any composition comprising a neuraminidase inhibitor and a peptide as defined above (of course also suitable (i.e., noninterfering interfering) mixtures of such substances) which prevents, ameliorates or heals the conditions described herein. In particular, the term " a pharmaceutical composition " to a composition comprising a neuraminidase inhibitor and a peptide as described above, and a pharmaceutically acceptable carrier or excipient (both terms can be used interchangeably). Suitable examples of carriers or excipients known to those skilled in the art are water, saline, sodium phosphate, sodium acetate, sodium carbonate, citrate, glycine, glycylglycine, histidine, lysine, arginine, TRIS and sodium citrate, or mixtures thereof. Of course, Ringer's solution, dextrose solution or solutions of non-reducible sugars can also be used; Accordingly, mannitol, trehalose, sucrose, sorbitol, fructose, maltose, lactose or dextran, Hank's solution, fixed oils, ethyl oleate, 5% dextrose in saline, substances that enhance isotonicity and chemical stability, buffers and preservatives are also suitable as such carriers. Other suitable carriers include any carrier that does not itself induce the production of antibodies harmful to the composition-sustaining individual, such as proteins, polysaccharides, polylactic acids, polyglycolic acids, polymeric amino acids, and amino acid copolymers. When formulating the composition according to the invention, of course, the relevant guidelines (for example the (European or US) Pharmacopoeia) must be observed. The peptide provided in the composition according to the invention can also be cyclized by direct covalent binding to these carriers.
The pharmaceutical composition according to the invention can be administered (as a medicament) with any suitable method known to those skilled in the art; in particular it is preferred to administer the peptide or composition according to the invention to the lung. Although influenza also plays a certain role in animals and the composition of the invention, of course, also for prevention and prophylaxis. Animals can be used, since the main focus of the present invention on the Voreuue tion and treatment in humans, aiso of persons who have already been infected with influenza or the laulc-n danger of becoming infected with this virus (especially in influenza Epidemics or pandemics). The preferred route of administration is * I * Φ Φ * Φ »* * Φ» »II« «* Φ« Φ Φ φ 4 ΦΦ ΙΦφ ··· »ΦΦΦ · φ Φ · φφ - 8 -
Inhalation (by aerosol) but also intravenous administration, instillation, oral administration or combinations thereof. By inhalation, parenteral or oral administration, the medicament of this invention is formulated in unit dosage form, such as a solution, suspension or emulsion, in association with the pharmaceutically acceptable excipients defined above. However, the dosage and mode of administration may of course also depend on the particular individual in the individual case.
In this case, the respectively required effective amount is administered to the individual who needs the administration. Where the "effective quantity" is as an amount effective enough to achieve the intended therapeutic or prophylactic effect, e.g. to prevent further spread of the disease or to treat it effectively. This is usually an average patient, but the actual effective levels of the components in the composition can be formulated to take into account the nature of the administration and the age, weight, condition of the patient and degree and progression of the disease (eg a suitable, conventional pharmacological protocol).
Preferably, therefore, the pharmaceutically acceptable carrier in the composition of the present invention is selected from water (more preferably: water for injection), saline, sodium phosphate, sodium acetate, sodium carbonate, citrate, glycine, glycylglycine, histidine, lysine, arginine, TRIS, sodium citrate, Ringer's solution, [dextrose, mannitol, trehalose, sucrose, sorbitol, fructose, maltese, lactose or dextran, Hank's solution, fixed oils, ethyl oleate, substances that improve isotonicity and steric stability, preservatives, pharmaceuticals acceptable proteins, polysaccharides, poly-lactic acids, polyglycolic acids, polymeric amino acids and amino acid copolymers.
The composition according to the invention can also be prepared by providing the two active components, ie the Neura-miridase inhibitor and the peptide, in a spatially separate form, as Ser, which is at least one neuraminidase inhibitor a peptide (each in separate containers). Accordingly, the present invention also relates to a kit comprising at least one novel rabbit inhibitor and one peptide {each in separate containers). The thereby enabled (spatially, but also temporally) separate administration of peptide and the inhibitor is especially preferred when different routes of administration of the two active components of the invention for the patient concerned are desired. For example, oselta-mivir is usually given orally, but the peptide to be used according to the invention is usually inhaled. However, concomitant administration is often desirable, for example zanamivir is also administered by inhalation. Of course, it may also be indicated that an oral inhibitor is provided systemically by inhalation. In some cases, the peptide to be used in the present invention is difficult to mix with the neuraminidase inhibitor, e.g. if the inhibitor is i.v. orally and the peptide is given by inhalation. In many cases, however, both inhibitor and peptide can be administered together by inhalation, with the inhibitor then entering directly into the lungs and possibly through the latter (influenza viruses are first present in the lung tissue).
Of course, in such a kit according to the invention comprising separate neuraminidase and peptide components, the features and preferred embodiments described herein for the mixed composition may of course also be provided in all combinations conceivable by those skilled in the art.
For example, the medicament according to the invention may be administered such that the peptide of the present invention is administered at a dose of between I μα / kg and 10 mg / kg, more preferably between 10 μg / kg and b mg / kg, most preferably between 0 , 1 and mg / kg. Preferably it is taken as a bolus dose. However, continuous inhalation or infusion or by repeated administration may also be used.
Particularly preferred compositions according to the invention are still present. the pg independently -
Peptide in an amount of 1 gg to 10 g, preferably 10 gg to 1 g, in particular from 1 mg to 100 mg, and inhibitor of the viral. Neuraminidase in an amount of 1 to 10 g, preferably from 100 gg to 1 g, in particular ago. 1 mg to 200 mg.
Particularly preferred compositions according to the invention in * »* * 0 0 0 * 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 * ·« ··· «·» ·· ·· ··· - 10 - liquid form independently contain the peptide in an amount of 1 pg to 10 g, preferably from 10 pg to 1 g, in particular from 1 mg to 100 mg, and the inhibitor of viral neuraminidase in an amount of 1 pg to 10 g , preferably from 100 pg to 1 g, in particular from 1 mg to 200 mg, and are in a volume of 0.5 to 10 ml, in particular in a volume of 1 to 5 ml, before.
The composition according to the invention may preferably also be administered in dry form by means of a powder inhaler. Examples of such powder inhalers which can be used in the present invention are described in U.S. Patents 4,995,385 and 4,069,819; already established products are SPINHALER · ', ROTAHALER®, FLOWCAPS®, INHALATOR®, DISKHALER® and AEROLIZER®.
The composition according to the invention can preferably also be administered as an aerosol by means of liquid nebulizer. Examples of such liquid nebulizers are well established
Products like Aeroneb® and Pari '".
According to a preferred embodiment, the composition according to the invention is characterized in that the peptide and / or the inhibitor of the viral neuraminidase are present in a nebulizable powder form or in a nebulizable liquid formulation.
The composition according to the invention is preferably used for the treatment and prophylaxis of infections with the influenza virus type A and type B, in particular type A. However, the composition is also suitable in principle, injections with. to prevent or treat any strain of the influenza virus that can cause this disease in an animal or in a human. Appropriate databases of information on the various types of influenza are familiar to those skilled in the art, and in particular many isolated type A strains are described or even sequenced.
The invention will be explained in more detail with reference to the following examples and the Zeichiangsiiguren to which, however, it is not limited.
Show: F'ig. 1: the relative maintenance of Cb7BL / 6 mice on days 3, 5, 7 and 9) infection with influenza A or (as 11-
Control) without infection;
Fig. 2: the relative lung weight of C57BL / 6 mice on days 5, 7 and 9 after infection with influenza A and treatment with oseltamivir, treatment with the composition according to the invention and without treatment (with BBS, as control).
Examples
The present examples demonstrate in a recognized experimental mouse model that the object of the present invention has been achieved by administering to influenza virus infected mice both a neuraminidase inhibitor and a combination of the neuraminidase inhibitor and the synthetic peptide AP301 (SEQ ID NO: 1) has been.
example 1
Infection with influenza virus causes the development of pneumonia.
Laboratory mice (strain C57BL / 6, 8 weeks old) were infected by nasal with influenza strain A (PR8 / 34) and a dosage of 150 PFU. On the 3rd, 5th, 7th and 9th day after the infection, the lungs were removed from each of δ mice and the relative lung weight was determined as a measure of pneumonia.
The investigation showed that with increasing duration after infection of the mice with the influenza virus, the lung weight. Compared with Kont roliiur.gen increased sharply. The results are shown graphically in FIG.
Example 2
Treatment of pneumonia by administration of Neurami -ni dashehe-r.mer or administration of a combination of neuraminidase inhibitor and peptide AP3G1.
Laccrrr.äuse (strain C5 bBL / 6, 3 weeks old) were nasally with the. Infiuasas ramm A (PR8 / 34) and a dosage of 150 PFU infected. Subsequently, the experimental animals each received oral doses of 10 m / g oseltamivir (neuraminidase inhibitor) and intratracheal doses: 10 μg / test animal peptide APJüi. The treatment was repeated on the 2nd and 4th day of testing.
At the. On the 5th, 7th, and 9th day after infection, the lungs were removed from each of 30 mice and the relative lung area was measured
* Weight determined as a measure of pneumonia. The results are shown graphically in FIG.
The study revealed that the neuraminidase inhibitor exerted only a modest effect in reducing pneumonia, as measured by lung weight. However, when the peptide AP301 was administered to the mice infected with influenza virus in addition to the neuraminidase inhibitor, the pneumonia could be reduced much more.
Example 3
Ex vivo assessment of the pro-inflammatory properties of the AP301 peptide in human whole blood.
A pharmacological ex vivo safety study of the AP301 peptide in human whole blood was performed to determine if the AP301 peptide was released from the fresh pro-inflammatory marker interleukin-6 (IL-6)
Whole blood (i.e., whether or not APN 301 exhibits TNF-specific inflammatory activity (i.e., TNF receptor binding activity). Fresh whole blood was used in this study, which is a recognized predictive model for the assessment of the in vivo depletion reaction.
Summary of the methodology
The objective of this study was to determine the lamphoric phosphorus signal capacity of the AP301 peptide. Whole blood cultures were users and secretion. I r.terleukin-6 (IL-6), a very sensitive marker of proinflammatory stimulation, quantified by ELISA.
Test, systern
Test system 25 ml of heparinized heparinized blood from 5 healthy volunteers <GP) was used in the tests.
Test object [dent i f cation:
Descriptive: AP3C1 peptide (dose: 1 ng / ml to 10 pq / rn I, one-time esterification in solution}
White powder, purity. 96%
Volcano Kui tures
* * * · · Λ * »· · · · · · · · · · · · ·
Whole blood (VB) cultures were performed by pipetting 1 ml of VB into wells of 24-well plates. In each experiment, unstimulated and stimulated control cultures were included.
Whenever possible, the substances and stimulants to be tested were always used in equal volume in each well in a given experiment, which is no greater than 10% of the total volume in a well. Unstimulated controls were done with PBS. Volume division and dilutions for various treatments were also performed with PBS.
The contents of each well were mixed and the plates incubated at 37 ° C and 5% CO2 for 24 hours. After incubation, the contents of each well were transferred to a fresh 1.5 ml microtube and centrifuged at 8000 to 9000 x g for 15 minutes. The supernatant from each sample was split individually into two 1.5 ml tubes and stored at -20 ° C until use.
Detection of interleukin-6
Interleukin-6 was quantitated by a specific ELISA (Human IL-6 ELISA Set, BD Biosciences, CaL No. 555220) using an anti-human i! 6 antibody as a capture antibody, a biotinylated anti-human IL-6 Deucleation antibody, avidin-merrettieh peroxidase conjugate as enzyme reagent and recombi nant.em I. i6 as standard. Absorption measurement at 450 nr. was introduced with dom Packard FusionRoadcr.
data analysis
The results of each plate were saved and evaluated with the Fusi onDataAnalysi s software.
Summary of the study results
The aim of this study was to determine the pro-inmammatory signaling capacity of the ΛΡ301 peptide. Whole blood cultures were used and secretion of IL-6, a highly sensitive marker of inflammatory Prc-St imuiar fon, was evaluated. - ELISA, quantify. the
Vliblut samples from five healthy volunteers were either left unstimulated (negative control), stimulating me. high and low doses of LPS (positive controls) or with 14-
* 4 4 · 4 4 f · f · f t 4 ♦ * ff
Peptide in nine ha.l blogarithm.iscr.en dilutions of 10 pg / ml to 1 ng / ml incubated. The results are shown in the following table:
Table: Release of Interleuk.in-6 from Fresh Whole Blood Upon Addition of Peptide AP301 and LPS AP301 Peptide Positive Control (LPS)
concentration
Concentration of IL-6 (pg / ml, n-5) 0 (negative control) 10 mg / ml 1 mg / ml 3 ng / ml 1 ng / ml less than 0.5 less than 0.5 less than 0.5 less 0.5 less than 0.5 less than 0.5 195.640 108.370 34.867 not determined
The results clearly show that the AP301 peptide did not induce a detectable level of IL-6 secretion at any of the concentrations tested. The positive controls (LPS) resulted in a strong induction of the IL-6 secretion ion.
discussion
The experiments were carried out to determine whether the AP301 peptide induced induction of pro-nflanur. Iter cascade mediated. Reacout parameter was the induced secretion of IL-6 in whole blood cultures from five healthy donors. The results clearly showed that the AP3C1 peptide had no detectable level of .11, -6 in the donor serum. induced. Thus, it has been demonstrated that the AP301 peptide does not induce nuclear pro-inflamatory response in the selected ex vivo-modoil and thus has no TNF receptor-binding activity.
Summary of the sequences:
SEQ ID NO. 1 CGQRETPEGAEAKPWYC SEQ ID NO: 2 KSPGGQRETPEGAEAKPWYE SEQ ID NO: 3 CGQREAPAGAAAKPWYC SEQ ID NO. 4 TPEGAE SEQ ID NO. SEQ ID NO: 7 CGQRETPEGAEAKPWYC SEQ ID NO: 9 CGQRETPEGAEARPWYC SEQ ID NO: 10 CGQRETPEGAEAKPC SEQ ID NO: 11 * »* · ·» · · · · · · · · «··· * · *» · «** · **« t · «I t 1
SEQUENCE LISTING < 110 > Apeptico Research and Development GmbH < 120 > Treatment and prevention of influenza < 130 > R 57719 < 160 > 12 < 170 > Patent version 3.5 < 210 > 1 < 211 > 17
&Lt; 212 > PRT < 213 > Artificial Sequence < 220 > &Lt; 223 > synthetic peptide < 400 > 1 1 5 cys < 210 > &Lt; 211 > &Lt; 212 > &Lt; 213 > 2 20 PRT Arti fi ci al Sequence < 220 > < 22 3 > Synthesis pepti de < 400 > 2
Cys Gly Gin Arg Glu Thr Pro Glu Gly Ala Glu Ala Lys Pro Trp Tyr
Lys ser Pro Gly Gly Gin Arg Glu Thr Pro Glu Gly Ala Glu Ala Lys 15 10 15
Pro Trp Tyr Glu < 210 > 20 3 < 211 > 17 < 212 > PRT < 213 > Arti fi ci al sequence < 220 > &Lt; 223 > Synthetic peptide < 400 > 3
Cys Gly Gin Arg Glu Ala Pro Ala Gly Ala Ala Ala Lys Pro Trp Tyr 15 10 15
Cys 2 < 210 > 4
&Lt; 211 > 6 < 212 > PRT < 213 > Artificial sequence < 220 > &Lt; 223 > synthetic peptide < 400 > 4
Thr Pro Glu Gly Ala Glu 1 5 < 210 > 5 < 211 > 14
&Lt; 212 > PRT < 213 > Artificial sequence < 220 > &Lt; 223 > synthetic peptide < 400 > 5
Gin Arg Glu Thr Pro Glu Gly Ala Glu Ala Lys Pro Trp Tyr 1 5 10 < 210 > 6 < 211 > 14
&Lt; 212 > PRT < 213 > Artificial Sequence < 220 > &Lt; 223 > synthetic peptide < 400 > 6
Pro Lys Asp Thr Pro Glu Gly Ala Glu Leu Lys Pro Trp Tyr 1 5 10 < 210 > 7 < 211 > 17
&Lt; 212 > PRT < 213 > Artificial Sequence < 220 > &Lt; 223 > synthetic peptide < 400 > 7
Cys Gly Pro Lys Asp Thr Pro Glu Gly Ala Glu Leu Lys Pro Trp Tyr 15 10 15
Cys < 210 > 8 < 211 > 17
&Lt; 212 > PRT < 213 > Artificial Sequence < 220 > synthetic peptide < 223 > 3 < 400 > 8 cys Gly Gin Lys Glu Thr Pro Glu Gly Ala Glu Ala Lys Pro Trp Tyr 15 10 15
Cys < 210 > 9 < 211 > 17 < 212 > PRT < 213 > Artificial sequence < 220 > < 22 B > Synthetic peptide < 400 > 9 1 5 Cys
Cys Gly Gin Arg Glu Thr Pro Glu Gly Ala Glu Ala Arg Pro Trp Tyr < 210 > 10 < 211 > 15 < 212 > PRT < 213 > Arti fi ci al sequence < 220 > &Lt; 223 > Synthetic peptide < 400 > 10 1 5 < 210 > 11 < 211 > 16 < 212 > PRT < 213 > Artificial Sequence < 220 > &Lt; 223 > synthetic peptide < 400 > 11
Cys Gly Gin Arg Glu Thr Pro Glu Gly Ala Glu Ala Lys Pro Cys
Cys Gin Arg Glu Thr Pro Glu Gly Ala Glu Ala Lys Pro Trp Tyr cys 15 10 15 < 210 > 12 < 211 > 17
&Lt; 212 > PRT < 213 > Artificial Sequence < 220 > &Lt; 223 > synthetic peptide < 400 > 12
Cys Gly Gin Arg Glu Thr Pro Glu Gly Ala Glu Ala Lys Phe Trp Tyr 15 10 15 »
* ♦ 4 cys

权利要求:
Claims (13)
[1]
Claims 1. A composition comprising a peptide consisting of 7-17 contiguous amino acids comprising the hexamer TX: EX2X3E, where Xlr X2 and X3 are any natural or not natural amino acid, which peptide has no TNF receptor binding activity and is cyclized, and an inhibitor of viral neuraminidase, particularly for use in the prevention and treatment of Inf luenza.
[2]
The composition of claim 1, wherein the peptide consists of 7-17 contiguous amino acids and the hexamer comprises TPEGAE.
[3]
The composition of claim 1 or 2 wherein the cyclized peptide is selected from a sequence of consecutive nucleic acids selected from the group consisting of QRET PEGAEAKPWY, PKDTPEGAELKPWY, CGQRETPEGAEAKPWYC, CGPKDTPHGA-ELKPWYC, and fragments of at least 7 amino acids thereof Hexamer TPEGAE consists.
[4]
Composition according to any one of Claims 1 to 3, characterized in that pneumonia caused by influenza is treated.
[5]
5. The composition according to any one of claims 1 to 4, characterized in that the peptide comprises the amino acid sequence CGQRkTRKGAEAKPWYC and is cyclized via the C residues,
[6]
6. Zusammensct tion according to one of claims i to 5, characterized in that the peptide is cyclized by a Disu 1 f i.dbrüc ko between the C-residues. Composition according to any one of Claims 1 to 6, characterized in that the viral neuraminidase inhibitor is Zar.amivir or Osoltair.ivir.
[7]
Composition according to any one of Claims 1 to 7, characterized in that it comprises a pharmaceutically acceptable carrier and is prepared as a pharmaceutical composition which is suitable for administration to humans.
[8]
Composition according to any one of Claims 1 to 8, characterized in that it comprises a pharmaceutically acceptable carrier selected from water, in particular water for injection, saline, sodium phosphate, sodium acetate, sodium carbonate, citrate, glycine, glycylglycine, histidine, lysine , Arginine, TRIS, sodium citrate, Ringer's solution, dextrose, mannitol, trehalose, sucrose, sorbitol, fructose, maltose, lactose or dextran, Hank's solution, fixed oils, ethyl oleate, substances that enhance iso-tonicity and chemical stability , Preservatives, pharmaceutically acceptable proteins, polysaccharides, polylactic acids, polyglycolic acids, polymeric amino acids and amino acid copolymers.
[9]
10. The composition according to any one of claims 1 to 9, characterized in that they - independently - the peptide in an amount of 1 pg to 10 g, preferably from 10 pg to 1 g, in particular from 1 to 100 mg 100 mg, and the Inhibitor of the oral neuraminidase in an amount of from 1 pg to 10 g, preferably from 100 μg to 1 g, in particular from 1 mg to 200 mg.
[10]
11. The composition according to any one of claims 1 to IC, characterized in that it in liquid form, vorli eqt and - independently - the peptide in an amount of 1 pg to 10 g, preferably from 10 pg to 1 g, in particular of 1 mg to 100 mg, and the inhibitor of the oral neuraminidase in an amount of 1 pg to 10 g, preferably from 100 pg to 1 g, in particular from 1 mg to 200 ng, and in a volume before, 0.5 to 10 ml, in particular in a volume of 1 to 5 ml, is present.
[11]
12. The composition according to any one of Ansprücne 1 to 11, characterized in that the peptide and / or the Hemm ο1-! viral neuraminidase in a nebulizable powder formulation or in a nebulizable liquid formulation. 18 - * * I fl • # ** * * r * • · · • t • t f
[12]
A kit comprising the following components: a peptide consisting of 7-17 contiguous amino acids and comprising the hexamer TX1EX2X3E, wherein XIr X2 and X3 may be any natural or non-natural amino acid, wherein the peptide has no TNF receptor binding activity and cyclized, and an inhibitor of viral neuraminidase, in particular for use in the prevention and treatment of influenza.
[13]
14. A kit according to claim 13, characterized in that the components are as defined in any one of claims 1 to 12.

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法律状态:
2016-07-15| MM01| Lapse because of not paying annual fees|Effective date: 20151118 |

优先权:

申请号 | 申请日 | 专利标题

ATA1908/2010A|AT510585B1|2010-11-18|2010-11-18|COMPOSITION COMPRISING A PEPTIDE AND AN INHIBITOR OF VIRAL NEURAMINIDASE|ATA1908/2010A| AT510585B1|2010-11-18|2010-11-18|COMPOSITION COMPRISING A PEPTIDE AND AN INHIBITOR OF VIRAL NEURAMINIDASE|

AU2011331891A| AU2011331891A1|2010-11-18|2011-11-15|Composition comprising a peptide and an inhibitor of viral neuraminidase|

US13/885,705| US10344055B2|2010-11-18|2011-11-15|Composition comprising a peptide and an inhibitor of viral neuraminidase|

EP11785277.2A| EP2640410B1|2010-11-18|2011-11-15|Composition comprising a peptide and an inhibitor of viral neuraminidase|

RU2013127402/15A| RU2596785C2|2010-11-18|2011-11-15|Composition containing peptide and virus neuraminidase inhibitor|

KR1020137011221A| KR101872218B1|2010-11-18|2011-11-15|Composition comprising a peptide and an inhibitor of viral neuraminidase|

PCT/AT2011/000462| WO2012065201A1|2010-11-18|2011-11-15|Composition comprising a peptide and an inhibitor of viral neuraminidase|

CN2011800554632A| CN103200955A|2010-11-18|2011-11-15|Composition comprising a peptide and an inhibitor of viral neuraminidase|

CA2817787A| CA2817787C|2010-11-18|2011-11-15|Composition comprising a peptide and an inhibitor of viral neuraminidase|

CN202011515592.2A| CN112717122A|2010-11-18|2011-11-15|Compositions comprising peptides and viral neuraminidase inhibitors|

BR112013011076A| BR112013011076A2|2010-11-18|2011-11-15|composition comprising a peptide and a viral neuraminidase inhibitor|

DK11785277.2T| DK2640410T3|2010-11-18|2011-11-15|COMPOSITION INCLUDING A PEPTIDE AND AN INHIBITOR OF VIRAL NEURAMINIDASE|

JP2013539088A| JP5970465B2|2010-11-18|2011-11-15|Composition comprising peptide and viral neuraminidase inhibitor|

MX2013005391A| MX2013005391A|2010-11-18|2011-11-15|Composition comprising a peptide and an inhibitor of viral neuraminidase.|

ES11785277.2T| ES2525758T3|2010-11-18|2011-11-15|Composition comprising a peptide and a viral neuraminidase inhibitor|

US16/418,265| US20190309021A1|2010-11-18|2019-05-21|Composition Comprising a Peptide and an Inhibitor of Viral Neuraminidase|

US16/892,658| US11161881B2|2010-11-18|2020-06-04|Composition comprising a peptide and an inhibitor of viral neuraminidase|

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