RU2434880C1 - Polypeptide demonstrating antibacterial activity and suitable for obtaining therapeutic medications for treatment of human bacterial infections - Google Patents

Abstract

FIELD: medicine.

SUBSTANCE: obtained is polypeptide of the following formula: U-X₁₉-X₂₀-X₂₁-X₂₂-X₂₃-X₂₄-X₂₅-X₂₆-X₂₇-X₂₈-X₂₉-X₃₀-X₃₁-X₃₂-Z, with U=X₁₈, X₁₇-X₁₈, X₁₆-X₁₇-X₁₈, X₁₅-X₁₆-X₁₇-X₁₈, X₁₄-X₁₅-X₁₆-X₁₇-X₁₈, X₁₃-X₁₄-X₁₅-X₁₆-X₁₇-X₁₈, X₁₂-X₁₃-X₁₄-X₁₅-X₁₆-X₁₇-X₁₈, X₁₁-X₁₂-X₁₃-X₁₄-X₁₅-X₁₆-X₁₇-X₁₈, X₁₀-X₁₁-X₁₂-X₁₃-X₁₄-X₁₅-X₁₆-X₁₇-X₁₈, X₉-X₁₀-X₁₁-X₁₂-X₁₃-X₁₄-X₁₅-X₁₆-X₁₇-X₁₈, X₈-X₉-X₁₀-X₁₁-X₁₂-X₁₃-X₁₄-X₁₅-X₁₆-X₁₇-X₁₈, X₇-X₈-X₉-X₁₀-X₁₁-X₁₂-X₁₃-X₁₄-X₁₅-X₁₆-X₁₇-X₁₈, X₆-X₇-X₈-X₉-X₁₀-X₁₁-X₁₂-X₁₃-X₁₄-X₁₅-X₁₆-X₁₇-X₁₈, X₅-X₆-X₇-X₈- X₉-X₁₀-X₁₁-X₁₂-X₁₃-X₁₄-X₁₅-X₁₆-X₁₇-X₁₈, X₄-X₅-X₆-X₇-X₈- X₉-X₁₀-X₁₁-X₁₂-X₁₃-X₁₄-X₁₅-X₁₆-X₁₇-X₁₈, X₃-X₄-X₅-X₆-X₇-X₈- X₉-X₁₀-X₁₁-X₁₂-X₁₃-X₁₄-X₁₅-X₁₆-X₁₇-X₁₈, X₂-X₃-X₄-X₅-X₆-X₇-X₈- X₉-X₁₀-X₁₁-X₁₂-X₁₃-X₁₄-X₁₅-X₁₆-X₁₇-X₁₈ or X₁-X₂-X₃-X₄-X₅-X₆-X₇-X₈- X₉-X₁₀-X₁₁-X₁₂-X₁₃-X₁₄-X₁₅-X₁₆-X₁₇-X₁₈; Z=X₃₃, X₃₃-X₃₄, X₃₃-X₃₄-X₃₅, X₃₃-X₃₄-X₃₅-X₃₆, X₃₃-X₃₄-X₃₅-X₃₆-X₃₇, X₃₃-X₃₄-X₃₅-X₃₆-X₃₇-X₃₈, X₃₃-X₃₄-X₃₅-X₃₆-X₃₇-X₃₈-X₃₉, X₃₃-X₃₄-X₃₅-X₃₆-X₃₇-X₃₈-X₃₉-X₄₀ or X₃₃-X₃₄-X₃₅-X₃₆-X₃₇-X₃₈-X₃₉-X₄₀-X₄₁; X₁=Ser; X₂=Glu; X₃=Thr; X₄=Arg; X₅=Pro; X₆=Val; X₇=Leu; X₈=Asn; X₉=Arg; X₁₀=Leu; X₁₁=Phe; X₁₂=Asp; X₁₃=Lys; X₁₄=Ile; X₁₅=Arg; X₁₆=Gln; X₁₇=Val; X₁₈=Ile; X₁₉=Arg; X₂₀=Lys; X₂₁=Phe; X₂₂=Glu; X₂₃=Lys; X₂₄=Gly; X₂₅=Ile; X₂₆=Lys; X₂₇=Glu; X₂₈=Lys; X₂₉=Ser; X₃₀=Lys; X₃₁=Arg; X₃₂=Phe; X₃₃=Phe; X₃₄=Asp; X₃₅=Gly; X₃₆=Leu; X₃₇=Leu; X₃₈=Ala; X₃₉=Phe; X₄₀=Arg; X₄₁=Lys.

EFFECT: invention makes it possible to obtain polypeptide, efficient in treatment and prevention of human bacterial infections.

8 cl, 2 dwg, 17 tbl, 6 ex

Description

The technical field to which the invention relates.

The present invention relates to biochemistry, in particular to new peptide compounds having a bactericidal effect. More specifically, the invention relates to the use of such compounds for the manufacture of medicaments for the prevention or therapeutic treatment of a bacterial or fungal infection of a person.

The prior art.

Since the middle of the last century, antibiotics have been widely used to fight bacterial infections, but the use of antibiotics is faced with the problem of the emergence of resistant strains of microorganisms. An increase in resistance to classical antibiotics was observed in a number of human pathogens: Mycobacterium tuberculosis, Enterococcus faecium, Pseudomonas aeroginosa, Staphylococcus aureus. Streptococcus pneumoniae. Salmonella typhi. In connection with this phenomenon, the search for new substances with antimicrobial activity is an urgent task. The discovery in the 1980s of antimicrobial peptides as key tools of the innate immunity system allows the development of antibiotics based on them with a fundamentally new mechanism of action that does not cause resistance in bacteria. Therefore, antimicrobial peptides can be an alternative to classical antibiotics (Giuliani et al., 2008, Cell. Mol. Life Sci., 65, 2450-2460).

Antimicrobial peptides were isolated from plants, insects, fish, amphibians, and mammals (Ganz, T., 2005, Comb. Chem. High Throughput Screen, 8, 209-217). These peptides vary in length, amino acid composition and structure, but most of them are small cationic amphipathic peptides with a wide spectrum of activity against gram-positive and gram-negative bacteria, yeast, fungi and enveloped viruses. Several human proteins and peptides also have antimicrobial activity and play an important role in the innate immunity system.

To date, three main groups of human antimicrobial peptides are known (Table 1. Amino acid sequences of defensins, histatins, and cathelicidin (LL-37)): defensins, histatin, cathelicidins (Contreras R., 2005, Biotechnology Letters, 27, 1337-1347) .

Defensins are cationic peptides containing three disulfide bonds between six cysteine residues and having a three-loop beta-folded spatial structure. Two classes of defensins were discovered in humans: alpha-defensins and beta-defensins. Alpha defensins are chemoattractants with respect to monocytes, dendritic cells and T-lymphocytes. They are an important component of the human immune system, protecting the body from infection and participating in the development of an adaptive (acquired) immune response. Beta-defensins are chemoattractants for dendritic and memory T cells via the CCR6 chemokine receptor (Tang Y.-Q. et al., 1999, Science 286, 498-502). The mouse model shows that human defensins administered intranasally enhance the acquired immune response by increasing the level of antigen-specific IgG and IgM in blood serum (Lillard JW et al., 1999, Proc. Natl. Acad. Sci.USA 96, 651- 656). Defensins exhibit high antibacterial, antifungal and antiviral activity against a wide range of microorganisms (Selsted M.E. et al., 1984, Infect. Immun. 46, 150-154). In in vitro experiments, the minimum concentration inhibiting the growth of microorganisms for most defensins is in the range of 0.5-10 μM (Risso A., 2000, J. Leukoc. Biol. 68, 785-792). These data prove that defensins are involved in the processes of innate and acquired immunity.

Histatins are small, cationic, histidine-enriched peptides found in human saliva (MacKay B.J. et al., 1984, Infect. Immun. 44, 688-694). These peptides are produced and secreted by the submandibular, sublingual and parotid glands (van derSpek J.C. et al., 1989, Am. J. Hum. Genet. 45: 381-387). Studies have shown that histatins have some bactericidal and, most importantly, fungicidal effect (Pollock J.J. et al., 1984, Infect. Immun. 44: 702_707). These peptides are part of the innate immunity system and play a key role in limiting oral infections. Histatins have a disordered structure in aqueous solutions and form an alpha helix in non-aqueous solvents.

The third group of human antimicrobial peptides contains only one peptide - cathelicidin LL-37. LL-37 is formed by proteolytic cleavage of the C-terminal portion of the hCAP18 protein, which is expressed in leukocytes, such as neutrophils, monocytes, NK cells, T cells, B cells, and in the epithelial cells of the skin, gastrointestinal and respiratory tract ( Gudmundsson GH et al., 1996, Eur. J. Biochem. 238, 325-332). Increased expression of LL-37 occurs in response to an inflammatory or infectious process (Frohm M., 1997, J. Biol. Chem. 272, 15258-15263). LL-37 has antimicrobial activity against gram-positive and gram-negative bacteria (Turner J. et al., 1998, Antimicrob. Agents Chemother., 42, 2206-2214), as well as the ability to bind and neutralize bacterial lipopolysaccharide (LPS) and protect against endotoxic shock during purulent infection (Bals R. et al., 1999, Infect. Immun. 67, 6084-6089). In addition, LL-37 is a chemoattractant for neutrophils, monocytes, mast cells and T-lymphocytes, stimulates wound healing and re-epithelization of the skin (Zanetti M., 2004, J. Leukoc. Biol. 75, 39-48). LL-37 consists of 37 amino acids, does not contain cysteine and has a linear structure. Like histatin, cathelicidin has a disordered structure in a hydrophilic environment and form an amphipathic alpha-helix in a hydrophobic medium.

The main mechanism of action of human antimicrobial peptides is based on the electrostatic binding of a positively charged peptide to negatively charged components of the outer membrane of the pathogen, which are represented by lipopolysaccharide (LPS) in gram-negative bacteria, polysaccharide (teichoic acid) in gram-positive bacteria and phospholipids. Following binding to the membrane, the incorporation of the peptide into the lipid bilayer of the bacterial cell leads to the formation of pores, to randomization of the lipid bilayer and, ultimately, to destruction of the microorganism (White et al. 1995, Curr. Opin. Struct. Biol. 5, 521-527 )

The antimicrobial peptide databases contain more than one and a half thousand substances (http://aps.unmc.edu/AP/main.php). to date, some of them are at different stages of clinical trials as drugs for the treatment and prevention of local and systemic infections (Gordon YJ et al., 2005, Curr Eye Res. July, 30 (7), 505-515), but the need to create new structures suitable for obtaining drugs based on them is only increasing. Since candidate peptides for use as medicines must not only possess the desired biological activity, but also be suitable for industrial production. On this side, the above peptides have some drawbacks: histatins exhibit greater fungicidal activity than bactericidal activity, and this limits their use, and defensins, due to their structure, are difficult to obtain, since isomeric products can form between the cysteine residues during the formation of three disulfide bonds. which do not have biological activity. At the same time, cathelicidin (LL-37) does not contain cysteines, therefore it can be obtained by chemical synthesis in high yield and has a powerful bactericidal effect. LL-37 is the closest to the claimed peptide in structure and antimicrobial properties. Detailed information on the properties of LL-37 and its fragments, as well as on the use of these peptides for the treatment of various bacterial infections of humans, can be found in patents WO 2009/001087, WO 2004/067025, WO 96/08508, RU 2346950.

The present invention solves the problem of expanding the arsenal of antimicrobial peptides suitable for the preparation of drugs based on them for the treatment and prevention of bacterial infections of humans.

Disclosure of the invention.

The problem is solved due to the structure of the peptide SE-41, which is a retro-sequence of the C-terminal part of the protein CAP 18, which has antimicrobial activity and has the following amino acid composition SEQ ID NO: 1: SETRPVLNRLFDKIRQVIRKFEKGIKEKSKRFFDGLLAFRK, as well as fragments of the peptide SE-41, containing from peptide SE-41 16 to 40 amino acids and selected from the group consisting of

respectively. The amino acid sequences of the claimed peptides are given in table.2.

Retro peptides are molecules that have a topochemical affinity for the original peptides, isomeric to natural peptides and differing from them in the opposite direction of the peptide bonds. Such molecules may possess the biological activity of the starting compounds. For the C-terminal fragment of the human CAP 18 protein, a computer model of the alpha helix of both direct and retro sequences indicates a high probability of the formation of an amphipathic structure under physiological conditions, which is a key factor for peptides to exhibit bactericidal activity.

In addition to the retro peptides themselves, the invention also encompasses retro peptide analogues and modified retro peptides. Analogues include peptides obtained by replacing one or several amino acids with homologous ones, as well as peptides that are retro-sequences of the C-terminal part of the CAP 18 protein of various mammals, for example, macaque, gibbon, orangutan, chimpanzee, rat, gorilla, mouse, etc. .d., the sequences of which can be found in public databases (http://au.expasy.org/cgi-bin/blast.pl). An additional aspect of the invention are peptides derived from SE-41 of SEQ ID NO: 1 and its fragments by replacing one or more homologous amino acids. The following pairs of amino acids are related to homologous ones; the choice of any amino acid from a pair does not lead to a noticeable change in the properties of the peptide: Ala / Ser, Val / Ile, Asp / Glu, Thr / Ser, Ala / Gly, Ala / Thr, Ser / Asn, Ala / Val, Ser / Gly, Tyr / Phe, Ala / Pro, Lys / Arg, Asp / Asn, Leu / Ile, Leu / Val, Ala / Glu, Asp / Gly (WO 2004/108752). Another aspect of the invention are modified peptides that contain amino acids in the D form and / or the alpha-amino group of the N-terminal amino acid is acetylated and the alpha-carboxyl group of the C-terminal amino acid is amidated. Such modifications can increase the resistance of peptides to the action of proteolytic enzymes.

Thus, taking into account all conditions, the formula of the claimed peptides can be represented in general form:

and where the amino acids constituting the polypeptide are independently selected from D or L forms. The technical result of the claimed invention is achieved due to such properties of the new peptide as: the ability to obtain a peptide using chemical synthesis, bactericidal activity, wound healing effect.

Since the claimed peptides are relatively small in size and contain from 16 to 41 amino acids, they can be obtained using chemical synthesis. In addition, the peptides are linear and do not contain cysteines, which also simplifies their production by chemical synthesis, since the possibility of the formation of inactive isomers upon closure of disulfide bonds is excluded.

Additional features of the claimed peptides, which provide a bactericidal effect, are a positive total charge and the ability to form an amphipathic alpha-helix in a non-aqueous environment. A computer analysis of the amino acid sequences of the peptides SE-41 and its closest known analogue LL-37, performed using a program on the antimicrobial peptide database website (http://aps.unmc.edu/AP/prediction/prediction_main.php), is given in table 3.

Table 3.
Properties of the peptides SE-41 and LL-37. Peptide LL-37 SE-41 The amount of hydrophobic residues 13 fifteen The amount of glycine and proline residues 3 3 Negatively Charged Amino Acids 5 5 Positive Charged Amino Acids eleven 13 Other amino acids 5 5 Hydrophobicity percentage 35% 36% Total charge +6 +8 Protein-binding Potential (Boman Index) 2.99 kcal / mol 3.08 kcal / mol The number of hydrophobic residues located on one surface 10 13 The ability to interact with lipid membranes + +

From the data presented in Table 3, it follows that the key parameters for the formation of the amphipathic alpha helix and the manifestation of the bactericidal properties of the peptide are the parameters, namely, the positive total charge, the percentage of hydrophobicity, and the Boman index (Boman, HG (2003) J.Inter. Med. 254 : 197-215) and the number of hydrophobic residues on one side of the surface of the molecule, better for the inventive peptide SE-41 SEQ ID NO: 1 compared with the known analogue LL-37.

An additional aspect of the invention is the bactericidal activity of the claimed peptides, shown in experiments using flow laser cytofluorimetry on bacteria Staphylococcus aureus (Wood46). For complete inhibition of the growth of these microorganisms in experiments in vitro, micromolar concentrations of the claimed peptides are required. In this case, the LD50 for SE-41 is 4.0 μg / ml, and for LL-37 - 4.3 μg / ml. In addition, bactericidal activity was shown for the following fragments of the peptide SE-41: SE37, IR20, IR24, LN31, LN35, characterized in that they have the amino acid sequence of SEQ ID NO: 5, SEQ ID NO: 20, SEQ ID NO: 90, SEQ ID NO: 83, SEQ ID NO: 151.

Additionally, the bactericidal properties of the claimed peptides, for example, peptides SE41, LN31, IR20, IR24, characterized in that they have the amino acid sequence of SEQ ID NO: 1, SEQ ID NO: 83, SEQ ID NO: 20, SEQ ID NO: 90, were confirmed in the classical bacteriological experiment of inhibiting the formation of colony forming units on solid nutrient media on Petri dishes. Also, the claimed peptides have a wound healing effect in experiments on a model of planar wound defect in laboratory mice at a concentration of 25 μg / ml. Thus, the set of essential features of the claimed peptides allows to achieve a technical result - to use new bactericidal peptides to obtain drugs for the treatment and prevention of bacterial infections of humans.

Examples of carrying out the invention.

Example 1. The synthesis of the peptide SE41 SEQ ID NO: 1:

Obtaining Fmoc-Lys (Boc) -P (I).

A hydroxyl group p-alkoxybenzyl polymer is washed with dimethylformamide, diethyl ether and dried on a glass filter. 300 mg of n-alkoxybenzyl polymer and 13.2 mg of dimethylaminopyridine (0.11 mmol) are suspended in a minimum volume of dimethylformamide. 515 mg (1.1 mmol) of Fmoc-Lys (Boc) -OH, 149 mg (1.1 mmol) of 1-hydroxybenzotriazole and 172 μl (1.1 mmol) of NN'-diisopropylcarbodiimide are dissolved in 5 ml of dimethylformamide, the solution is stirred 10 min at 0 ° C and then added to the suspended polymer. The reaction is carried out for 4 hours with periodic stirring. At the end of the reaction, the polymer is filtered off, washed with dimethylformamide and treated with 5 ml of a mixture of Ac ₂ O-pyridine-CH ₂ Cl ₂ (20:20:60) for 1 h, after which the polymer is washed with isopropanol and dimethylformamide.

Preparation of Fmoc-Arg (Pbf) -Lys (Boc) -P (II).

a) The peptidyl polymer (I) obtained in the previous step for 20 minutes treated with a 20% solution of piperidine in dimethylformamide. The polymer is washed successively with 5 ml of the following solvents: dimethylformamide - 3 times for 2 minutes, a mixture of dioxane-water (2: 1) - 2 times for 5 minutes, dimethylformamide - 5 times for 2 minutes.

b) 713 mg (1.1 mmol) of Fmoc-Arg (Pbf) -OH, 149 mg (1.1 mmol) of 1-hydroxybenzotriazole and 172 μl (1.1 mmol) of NN'-diisopropylcarbodiimide are dissolved in 5 ml of dimethylformamide, solution stirred for 10 min at 0 ° C and then added to the suspended polymer. The reaction is carried out for 4 hours with periodic stirring. At the end of the reaction, the polymer is filtered off, washed with dimethylformamide and treated with 5 ml of a mixture of Ac ₂ O-pyridine-dimethylformamide (20:20:60) for 1 h, after which the polymer is washed successively with dimethylformamide, isopropanol, dimethylformamide.

Peptide synthesis SE41: SETRPVLNRLFDKIRQVIRKFEKGIKEKSKRFFDGLLAFRK.

The synthesis of the polypeptide chain is carried out manually in a glass flow reactor (2 × 20 cm) according to the following protocol for each synthetic cycle (based on 8-10 ml of solvent per 300 mg of the starting polymer); during the condensation reaction (step 6), the volume of the reaction mixture 3 -4 ml:

1. DMF (5 × 2 min);

2. 20% piperidine in DMF (20 min);

3. DMF (3 × 2 min);

4. dioxane-water, 2: 1, (2 × 5 min);

5. DMF (5 × 2 min);

6. Condensation reaction: 10 eq. activated Fmoc amino acids (4 hours);

7. DMF (3 × 2 min);

8. acetylation: Ac ₂ O-pyridine-dimethylformamide, 20:20:60, (1 h);

9. DMF (3 × 2 min);

10. isopropanol (3 × 2 min);

To activate the Fmoc-derived amino acids with the DIPCDI / HOBT method, 172 μl (10 equiv., 10 equivalents) are added to a solution of 10 eq. (1.1 mmol) of the Fmoc-protected amino acid and 149 mg (10 eq., 1.1 mmol) of HOBt in 3-4 ml of DMF. 1.1 mmol) DIPCDI, the solution is stirred for 10 minutes.

The completeness of the condensation reaction is monitored using the ninhydrin or, in the case of the N-terminal proline, picrin tests after step 6 of the synthetic protocol.

For the reaction of peptide cleavage from the polymer and simultaneous release of the protective groups of the amino acid side chains, 300 mg of the peptidyl polymer (from 1.2 g of the peptidyl polymer) are selected. 5 ml of a mixture of TFA-EDT-DMS-m-cresol are added to the peptidyl polymer in a volume ratio of 91: 3: 3: 3, the suspension is stirred for 2 hours, then the resulting peptide solution is filtered off from the polymer, washed with 5 ml of TFA and an excess of TFA evaporated under reduced pressure. The peptide is precipitated with 100 ml of ethyl ether, filtered and washed with ether (5 × 20 ml). The precipitate was dissolved in 5 ml of 10% AcOH for 20 minutes, filtered and washed with 5 ml of 10% AcOH. The resulting peptide solution was lyophilized and desalted on a column (2.5 × 60 cm) with Sephadex G-10 in 0.1 M AcOH. The peptide was purified using reverse-phase HPLC in an acetonitrile gradient (from 10% to 70% in 60 min) in 0.1% TFA at an eluent flow rate of 4 ml / min, the absorption of the eluate was recorded at a wavelength of 226 nm. Fractions corresponding to the main peak in the chromatogram are collected and lyophilized. The yield of the peptide based on the C-terminal amino acid was 30%. The peptide is analyzed by analytical HPLC and mass spectrometry. Analytical reverse-phase HPLC was carried out in an acetonitrile gradient in 0.1% TFA (from 10% to 70% in 60 minutes) at an eluent flow rate of 1 ml / min, the absorption of the eluate was recorded at a wavelength of 226 nm. The retention time of the peptide under analytical HPLC was 40.3 minutes. Experimental molecular weight 4993.5, theoretical molecular weight 4992.9.

Abbreviations:

DIPCDI-N, N'-diisopropylcarbodiimide; Fmoc-9 - fluorenylmethyloxycarbonyl; NOVT - 1-hydroxybenzotriazole; TFA - trifluoroacetic acid; DMAP - dimethylaminopyridine; EDT - ethanedithiol; DMS Dimethyl Sulfide; AcOH - acetic acid; DMF - dimethylformamide

Example 2. Evaluation of the bactericidal activity of the peptides LL-37 and SE-41 by flow laser cytofluorimetry.

The antimicrobial properties of synthetic cationic peptides (LL37 and SE41) are investigated in the St. model. aureus (Wood46).

The daily agar culture of St.aureus (strain Wood46) was washed with phosphate-buffered saline (PBS), disintegrated using ultrasound on an ultrasonic bath (51 kHz, 90 W) for 2 hours, precipitated at 1000 g for 25 minutes and washed with 10 ml of PBS. The second time the washing is carried out with 0.01 M carbonate-bicarbonate buffer (pH = 9.5). Resuspend in the same buffer and, according to the turbidity standard, the concentration of bacteria is adjusted to 1 × 10 ⁸ / ml. A freshly prepared solution of fluorescein isothiocyanate - FITC (Sigma) is added to the suspension at the rate of 1 mg / ml and the carbonate-bicarbonate buffer is kept at pH = 9.5 for 16 hours at + 4 ° С in the dark (St.aureus-FITC ⁺ ). After incubation, St.aureus-FITZ ⁺ was washed three times with FSB (1000g 25 min), resuspended in FSB with the addition of 5% fetal calf serum and 5% DMSO. The concentration of St.aureus-FITZ ^{+ was} adjusted to 5 × 10 ⁸ / ml using Tru Count tubes (Becton Dickinson) with a known number of microbeads. Aliquots are stored at -70 ° C.

After defrosting, aliquots of bacteria are precipitated and washed in a 10 mM FSB solution pH = 7.4 2 times at 1000g 25 min.

The reaction of the bactericidal activity of biological fluids is carried out in a 96-well round-bottom plate. To do this, incubate 90 μl of cationic peptides and 90 μl of St.aureus-FITC ⁺ (1 × 10 ⁷ CFU / ml) for 3 hours at + 37 ° C. The control of the reaction is the spontaneous death of St.aureus-FITC ⁺ (bacteria with FSB under the same conditions).

After incubation, propidium iodide (2.5 μg / ml) is added to the bacteria to the FSB. As you know, propidium iodide penetrates and stains only dead cells. After 10 minutes, the samples were analyzed on a FACSCalibur flow cytometer (Becton Dickinson) in the CellQuest program at coordinates FL1 and FL3. Among the general population of staphylococcus cells stained with FITC (green fluorochrome), the percentage of staphylococcus cells stained with propidium iodide (red fluorochrome) is determined (Table 4. Antibacterial activity of synthetic cathelicidins). Next, using the Exel program, LD50 is calculated by the Ashmarin-Kerber method (Table 5. Determination of LD50 of preparations in relation to St.aureus Wood46).

Table 4.
Antibacterial activity of the peptides LL-37 and SE-41. Experience number Percentage of death of St.aureus LL37 mcg / ml SE41 mcg / ml 0 one 10 one hundred 0 one 10 one hundred one one 5 70 83 2 6.4 67.4 78 2 one 5 70 84 2 6 82 74 3 one 4.9 73 86 2 7 88.9 82 four 1.98 11.98 76.7 88.5 3 7 81.7 71.2 5 1.98 eleven 88.68 91.5 3 7.2 81.5 75.9 6 1.98 12 79.7 94.5 3 8 87.8 73.8

Table 5.
Determination of LD50 of peptides LL-37 and SE-41 against St.aureus Wood46. Experience number LD50, mcg / ml LL37 SE41 one 5,0 5,4 2 5,0 3.9 3 4.7 3.4 four 4.0 3.9 5 3.3 3.9 6 3.8 3,5 Avg. 4.3 4.0 Standard deviation 0.7 0.7

Example 3. Evaluation of the antimicrobial effect of synthetic cationic peptides LL37 and SE41 using the classical bacteriological method.

The antimicrobial effect of the obtained synthetic cationic peptides (cathelicidins) was evaluated by the classical bacteriological method. The antimicrobial properties of synthetic cationic peptides (LL37 and SE41) were studied on the models of St.aureus (Wood46) and Escherichia coli (ATCC 25922).

Bacterial culture, grown on beveled trypticosoy soy agar (Beckton Dickinson), washed with 10 mM PBS (Sigma) saline pH = 7.4 and precipitated at 1000g for 25 minutes. It is then washed in PBS and the precipitate is resuspended in PBS. The method of seeding on solid nutrient media determines in three independent experiments the number of colony forming units (CFU) in 1 ml. The optical density (OD) of the bacterial suspension was measured on a Beckman Coulter DU 730 spectrophotometer at λ = 620 nm. OD _{620 of} 0.2 corresponds to 5 × 10 ⁷ CFU / ml. The concentration of bacteria in all experiments adjusted PBS to 1 × 10 ⁷ CFU / ml

The experiment is carried out in 1.5 ml sterile plastic tubes (Sarstedt). 10 μl of bacteria, 10 μl of a solution of a synthetic peptide in an appropriate concentration and 30 μl of PBS are added to the tubes. The peptide was not added to the control tube. When evaluating the antibacterial effect of peptides in the St. aureus and E. coli use concentrations of 0.2, 2, and 20 μg / ml and 2.5, 5, 10, and 20 μg / ml, respectively. Plastic tubes with a mixture of bacteria and peptides are incubated in a TW-2.02 water bath (ELMI) for 3 hours at 37 ° C. After incubation, the mixtures were diluted with PBS and inoculated in a volume of 50 μl on trypticase soy agar (Beckton Dickinson) in Petri dishes. Cups are placed in a thermostat at 37 ° C for 18-24 hours, after which the grown colonies are counted. Determine the percentage of bacteria killed in relation to the control and calculate LD ₅₀ according to the method of Reed and Mench.

The experimental results are presented in table.6. Evaluation of the antimicrobial effect of drugs on the St.aureus model (Wood46) and Table 7. Evaluation of the antimicrobial effect of drugs on the E. coli model (ATCC 25922), as well as in figure 1. The effect of suppressing the growth of a bacterial culture (St. aureus Wood 46) and Fig.2. The effect of suppressing the growth of bacterial cultures on the E. coli model (ATCC 25922). The experimental results show that both peptides LL-37 and SE-41 have high bactericidal activity against St. aureus and E. coli, and in low concentrations the effect on E. coli of the peptide SE-41 is higher than LL-37.

Table 6.
Evaluation of the antimicrobial effect of drugs on the St.aureus model (Wood46). Peptide Dose (mcg / ml) 0 0.2 2 twenty colonies colonies colonies colonies No. number % killed number % killed number % killed number % killed LL37 one 67 0 56 16,4 thirty 46,4 3 99 2 67 0 55 17.9 31 43.6 5 98.4 3 67 0 fifty 25,4 thirty 40,0 2 99.3 M ± δ 67 ± 0 0 ± 0 53.7 ± 3.2 19.9 ± 4.8 30.3 ± 0.6 43.4 ± 3.2 3.3 ± 1.5 98.9 ± 0.5 SE41 one 70 0 66 5.8 34 47.6 3 91.1 2 70 0 60 14,4 33 45,2 0 one hundred 3 70 0 61 13,2 33 45.6 four 98.8 M ± δ 70 ± 0 0 ± 0 62.3 ± 3.2 11.1 ± 4.7 33.3 ± 0.6 46.1 ± 1.3 2.3 ± 2.1 99.3 ± 0.6

Table 7.
Evaluation of the antimicrobial effect of drugs on the E. coli model (ATCC 25922). Peptide 0 2.5 10 twenty colonies colonies colonies colonies LL37 No. number % killed number % killed number % killed number % killed one 42 0 36 fourteen 17 59.5 2 95 2 thirty 0 24 twenty four 86.7 2 93 3 thirty 0 24 twenty one 96.7 2 93 M ± δ 34 ± 7 0 ± 0 28 ± 7 18 ± 3 7.3 ± 8.5 81 ± 19 2 ± 0 94 ± 1 SE41 one thirty 0 24 twenty 3 90 2 93.3 2 31 0 25 19 2 83.3 2 93.3 3 thirty 0 twenty 33 5 93.5 2 93.5 M ± δ 30 ± 0.5 0 ± 0 23 ± 2 24 ± 6 3 ± 1.2 89 ± 4 2 ± 0 93 ± 1

Table 8.
Determination of LD50 peptides LL-37 and SE-41 on the model St. aureus (Wood 46). Experience number LD50, mcg / ml LL37 SE41 one 1,5 1.9 2 1,6 1.7 3 1.3 1.7 mean 1,5 1.8 senior off 0.2 0.1

Table 9.
Determination of LD50 of peptides LL-37 and SE-41 on the E. coli model (ATCC 25922). Experience number LD50, mcg / ml LL37 SE41 one 4,5 3,5 2 5.3 3,5 3 4.2 3,1 mean 4.7 3.4 senior off 0.6 0.2

Example 4. The study of the wound healing effect of the peptide SE41 on a model of planar wound defect in laboratory mice.

The wound healing effect of SE41 was evaluated on a full-layer wound in 20 F ₁ mice (CBA × C57 BL / 6). Under light ether anesthesia, the animals are shaved on their backs, a “mark” of 0.5 cm × 0.5 cm is applied on the screen and a full-layer skin flap is cut, simulating a standard wound defect in area. A day after wounding, animals are divided into 4 groups of 5 mice:

1 g - control mice, which saline is dripped onto the wound.

2 gr. - mice that are dripped on the wound with a solution of SE41 at a concentration of 100 μg / ml.

3 gr. - mice that are dripped on the wound with a solution of SE41 at a concentration of 50 μg / ml.

4 gr. - mice that are dripped on the wound with a solution of SE41 at a concentration of 25 μg / ml.

Animals of each group are kept in separate cages in the same conditions for 5 pcs. with free access to water and food.

For the preparation of working solutions SE41 using sterile pyrogen-free water with the addition of 0.9% sterile sodium chloride. Saline thus prepared serves as the basis for further dilution of SE41 to working concentrations. SE41 solutions are prepared before application to wounds.

SE41 solutions are dripped into the wound one day after the injury with the help of an insulin syringe without a needle of 0.1 ml, then daily for 7 days.

On days 2, 4, 8, and 11 from the start of the experiment, the size of the wounds is measured by the mass of mica patterns. The measurement results are presented in table 10-14.

Table 10.
Change in the size of the wound surface in mice of the control group. Animal number Pattern weight (mg) / day of observation 2 four 8 eleven fourteen fifteen one 6.5 5,0 2.5 1,5 0 2 6.0 5,0 2.0 1,0 0 3 6.5 5,0 2.5 1,5 0 four 6.0 5,0 1,5 1,0 0 5 6.0 5,0 1,5 0 Average 6.2 5,0 2.0 1,0 0

The results of the observation show that in the control group of mice, without using the SE41 preparation on the wound, the wounds healed by the 14th day, one mouse out of 5 had no wound defect on the 11th day after the injury.

Table 11.
Change in the size of the wound surface in mice of group No. 2 Animal number Pattern weight (mg) / day of observation 2 four 8 eleven fourteen fifteen one 6.0 6.0 3,5 1,5 0.5 0 2 6.0 5.5 4.0 1,0 0 0 3 6.5 6.0 4,5 2.0 0.5 0 four 6.0 6.0 3,5 1,0 0 0 5 6.0 6.0 3,5 1,5 0.5 0 Average 6.1 5.9 3.8 1.4 0.3 0

It should be noted that when applying the SE41 solution at a concentration of 100 μg / ml, the animals showed motor anxiety, after which they began to lick each other's wound surface. Subsequently, wounds in animals of this group externally differed from wounds in animals of other groups. 48 hours after the injury in animals of the control group, No. 3 and No. 4, the wound defect was a dry scab (crust), in animals, gr. No. 2 within 4 days after the injury, the wound was dry, but no crust formed, the edges of the wound seemed to be raised above the bottom of the wound. Then a dry scab formed and the appearance did not differ anymore, but the sizes were on average larger, the complete marginal epithelization ended by 15 days.

Table 12.
Change in the size of the wound surface in mice of group No. 3 Animal number Pattern weight (mg) / day of observation 2 four 8 eleven fourteen fifteen one 6.0 5,0 2.0 1,5 0.5 2 6.0 5,0 2.0 1,0 0.5 3 6.5 5,0 1,5 1,5 0 four 6.0 4,5 1,5 1,0 0 5 6.5 5,0 2.0 1.5 0.5 Average 6.2 4.9 1.8 1.3 0.3

In group No. 3, the results of measurements of the wound defect did not practically differ from the control group, at the same time, a certain tendency to accelerate the regeneration process by 4-8 days after the wound should be noted.

In group No. 4, in the treatment of wounds with a solution of SE41 at a concentration of 25 μg / ml, acceleration of wound healing was revealed in all mice.

Table 13.
Change in the size of the wound surface in mice of group No. 4. Animal number Pattern weight (mg) / day of observation 2 four 8 eleven fourteen fifteen one 6.5 5,0 2.0 0.5 2 6.0 4,5 1,5 0 3 6.5 5,0 2.0 0.5 four 6.0 5,0 1,5 0 5 6.5 4,5 1,5 0 Average 6.3 4.8 1.3 0.5

Table 14.
Summary data on the wound healing effect of SE41 (masses Dose (mcg / ml) Observation days 2 four 8 eleven fourteen fifteen 0 6.2 5,0 2.0 1,0 0 0 25 6.3 4.8 1.3 0.5 0 0 fifty 6.2 4.9 1.8 1.3 0.3 0 one hundred 6.1 5.9 3.8 1.4 0.3 0 patterns in mg).

Thus, according to the results of this experiment, we can conclude that the peptide SE41 has wound healing activity when applied locally at a concentration of 25-50 μg / ml. The best effect is observed when using a solution of SE41 with a concentration of 25 μg / ml.

Example 5. Evaluation of the bactericidal activity of fragments of the peptide SE-41 by flow laser cytofluorimetry.

Evaluation of the bactericidal activity of the peptides is carried out as described in example 2. The results of the determination of LD50 are given in table 15. LD50 of SE-41 peptide and its fragments against St.aureus (Wood 46) in flow cytometry.

Table 15. LD50 of SE-41 peptide and its fragments against St.aureus (Wood 46) in flow cytometry. SEQ ID NO: Title LD50, mcg / ml SEQ ID NO: 1 SE41 5.0 + 0.5 SEQ ID NO: 5 SE37 3.9 + 0.4 SEQ ID NO: 20 IR20 3.8 + 0.6 SEQ ID NO: 26 IR16 4.1 ± 0.1 SEQ ID NO: 90 IR24 44.8 + 13.1 SEQ ID NO: 83 Ln31 9.9 + 3.8 SEQ ID NO: 151 Ln35 8.5 + 1.2

Example 6. Evaluation of the antimicrobial action of the peptide SE41 and its fragments using the classical bacteriological method.

An experiment to evaluate the antimicrobial effect of the SE41 peptide and its fragments is carried out as described in example 3. The results are shown in table.16. Evaluation of the antimicrobial effect of the peptide SE-41 and its fragments on the model of St.aureus (Wood46).

Table 16. Evaluation of the antimicrobial effect of the peptide SE-41 and its fragments on the model of St.aureus (Wood46). Peptide Dose (mcg / ml) 0 2.5 5 10 colonies colonies colonies colonies Experience number number % killed number % killed number % killed number % killed SE41 one 64 0 36 43.7 four 91.3 0 one hundred 2 64 0 31 51.6 2 94.3 0 one hundred 3 64 0 31 51.6 2 95.0 0 one hundred M ± δ 64 ± 0 0 ± 0 32.7 ± 2.9 49.0 ± 4.5 2.7 ± 1.2 93.5 ± 2.0 0 ± 0 100 ± 0 Ln31 one 63 0 32 49.2 12 81.0 0 one hundred 2 63 0 31 50.8 eleven 82.5 0 one hundred 3 63 0 28 55.6 6 90.5 0 one hundred M ± δ 63 ± 0 0 ± 0 30.3 ± 2.1 51.9 ± 3.3 9.7 ± 3.2 84.7 ± 5.1 0 ± 0 100 ± 0 IR20 one 57 0 fourteen 75.4 2 96.5 2 96.5 2 46 0 fourteen 69.6 one 97.8 0 one hundred 3 46 0 fourteen 69.6 one 97.8 0 one hundred M ± δ 49.7 ± 6.4 0 ± 0 14.0 ± 0 71.5 ± 3.4 1.3 ± 0.6 97.4 ± 0.8 0.7 ± 1.2 98.8 ± 0.8 IR24 one 65 0 65 0 27 58.5 3 95.4 2 65 0 60 7.7 12 71.5 2 96.9 3 65 0 fifty 23.1 7 89.2 one 98.5 M ± δ 65 ± 0 0 ± 0 58.3 ± 7.6 10.3 ± 11.8 15.3 ± 10.4 76.4 ± 16.0 2.0 ± 1.0 96.9 ± 1.5 IR16 one 67 0 55 eighteen 24 60 19 71.6 2 63 0 54 fourteen 22 63 16 74.6 3 58 0 53 9 21 65 fourteen 76 M ± δ 63 ± 5 54 ± 1 14 ± 5 22 ± 2 63 ± 3 16 ± 3 74 ± 2

Table 17. LD50 peptide SB-41 and its fragments against St.aureus (Wood 46) LD50, mcg / ml SE41 / SEQ NO NO: 1 LN31 / SEQ ID NO: 83 IR20 / SEQ ID NO: 20 IR24 / SEQ ID NO: 90 IR16 / SEQ ID NO: 26 one 2.7 2.5 1.7 4,5 4.0 2 2.1 2.5 1.8 3,7 4.1 3 2,3 2,3 1,2 3.3 4.2 Wed meaning 2,4 2,4 1,6 3.9 4.1 Art. off 0.3 0.1 0.2 0.6 0.1

Amino acid sequence of the peptide SE41 SEQ ID NO: 1:

SETRPVLNRLFDKIRQVIRKFEKGIKEKSKRFFDGLLAFRK

Claims (8)

1. The polypeptide having antibacterial activity and consisting of amino acids according to the following formula:
UX ₁₉ -X ₂₀ -X ₂₁ -X ₂₂ -X ₂₃ -X ₂₄ -X ₂₅ -X ₂₆ -X ₂₇ -X ₂₈ -X ₂₉ -X ₃₀ -X ₃₁ -X ₃₂ -Z, where U = X ₁₈ , X ₁₇ -X ₁₈ , X ₁₆ -X ₁₇ -X ₁₈ , X ₁₅ -X ₁₆ -X ₁₇ -X ₁₈ , X ₁₄ -X ₁₅ -X ₁₆ -X ₁₇ -X ₁₈ , X ₁₃ -X ₁₄ -X ₁₅ -X ₁₆ -X ₁₇ -X ₁₈ , X ₁₂ -X ₁₃ -X ₁₄ -X ₁₅ -X ₁₆ -X ₁₇ -X ₁₈ , X ₁₁ -X ₁₂ -X ₁₃ -X ₁₄ -X ₁₅ -X ₁₆ -X ₁₇ -X ₁₈ , X ₁₀ -X ₁₁ -X ₁₂ -X ₁₃ -X ₁₄ -X ₁₅ -X ₁₆ -X ₁₇ -X ₁₈ , X ₉ -X ₁₀ -X ₁₁ -X ₁₂ -X ₁₃ -X ₁₄ - X ₁₅ -X ₁₆ -X ₁₇ -X ₁₈ , X ₈ -X ₉ -X ₁₀ -X ₁₁ -X ₁₂ -X ₁₃ -X ₁₄ -X ₁₅ -X ₁₆ -X ₁₇ -X ₁₈ , X ₇ -X ₈ -X ₉ -X ₁₀ -X ₁₁ -X ₁₂ -X ₁₃ -X ₁₄ -X ₁₅ -X ₁₆ -X ₁₇ -X ₁₈ , X ₆ -X ₇ -X ₈ -X ₉ -X ₁₀ -X ₁₁ -X ₁₂ -X ₁₃ -X ₁₄ -X ₁₅ -X ₁₆ -X ₁₇ -X ₁₈ , X ₅ -X ₆ -X ₇ -X ₈ -X ₉ -X ₁₀ -X ₁₁ -X ₁₂ -X ₁₃ -X ₁₄ - X ₁₅ -X ₁₆ -X ₁₇ -X ₁₈ , X ₄ -X ₅ -X ₆ -X ₇ -X ₈ -X ₉ -X ₁₀ -X ₁₁ -X ₁₂ -X ₁₃ -X ₁₄ -X ₁₅ -X ₁₆ -X ₁₇ -X ₁₈ , X ₃ -X ₄ -X ₅ -X ₆ -X ₇ -X ₈ -X ₉ -X ₁₀ -X ₁₁ -X ₁₂ -X ₁₃ -X ₁₄ -X ₁₅ -X ₁₆ -X ₁₇ -X ₁₈ , X ₂ -X ₃ -X ₄ -X ₅ -X ₆ -X ₇ -X ₈ -X ₉ -X ₁₀ -X ₁₁ -X ₁₂ -X ₁₃ -X ₁₄ -X ₁₅ -X ₁₆ - X ₁₇ -X ₁₈ and whether X ₁ -X ₂ -X ₃ -X ₄ -X ₅ -X ₆ -X ₇ -X ₈ -X ₉ -X ₁₀ -X ₁₁ -X ₁₂ -X ₁₃ -X ₁₄ -X ₁₅ -X ₁₆ -X ₁₇ -X ₁₈ ; Z = X ₃₃ , X ₃₃ -X ₃₄ , X ₃₃ -X ₃₄ -X ₃₅ , X ₃₃ -X ₃₄ -X ₃₅ -X ₃₆ , X ₃₃ -X ₃₄ -X ₃₅ -X ₃₆ -X ₃₇ , X ₃₃ - X ₃₄ -X ₃₅ -X ₃₆ -X ₃₇ -X ₃₈ , X ₃₃ -X ₃₄ -X ₃₅ -X ₃₆ -X ₃₇ -X ₃₈ -X ₃₉ , X ₃₃ -X ₃₄ -X ₃₅ -X ₃₆ -X ₃₇ -X ₃₈ -X ₃₉ -X ₄₀ or X ₃₃ -X ₃₄ -X ₃₅ -X ₃₆ -X ₃₇ -X ₃₈ -X ₃₉ -X ₄₀ -X ₄₁ ;
X ₁ = Ser;
X ₂ = Glu;
X ₃ = Thr;
X ₄ = Arg;
X ₅ = Pro;
X ₆ = Val;
X ₇ = Leu;
X ₈ = Asn;
X ₉ = Arg;
X ₁₀ = Leu;
X ₁₁ = Phe;
X ₁₂ = Asp;
X ₁₃ = Lys;
X ₁₄ = Ile;
X ₁₅ = Arg;
X ₁₆ = Gln;
X ₁₇ = Val;
X ₁₈ = Ile;
X ₁₉ = Arg;
X ₂₀ = Lys;
X ₂₁ = Phe;
X ₂₂ = Glu;
X ₂₃ = Lys;
X ₂₄ = Gly;
X ₂₅ = Ile;
X ₂₆ = Lys;
X ₂₇ = Glu;
X ₂₈ = Lys;
X ₂₉ = Ser;
X ₃₀ = Lys;
X ₃₁ = Arg;
X ₃₂ = Phe;
X ₃₃ = Phe;
X ₃₄ = Asp;
X ₃₅ = Gly;
X ₃₆ = Leu;
X ₃₇ = Leu;
X ₃₈ = Ala;
X ₃₉ = Phe;
X ₄₀ = Arg;
X ₄₁ = Lys,
and where the amino acids constituting the polypeptide are independently selected from D or L forms. 2. The polypeptide according to claim 1, selected from the group consisting of: SE41, SE40, SE39, SE38, SE37, SE36, SE35, SE34, SE33, ET32, TR31, RP30, PV29, VL28, LN27, NR26, RL25, KF24 , FD23, DK22, KI21, IR20, RQ19, QV18, VI17, IR16, ET33, TR32, RP31, PV30, VL29, LN28, NR27, RL26, LF25, FD24, DK23, KI22, IR21, RQ20, QV19, VI18, IR17 , ET34, TR33, RP32, PV31, VL30, LN29, NR28, RL27, LF26, FD25, DK24, KI23, IR22, RQ21, QV20, VI19, IR18, ET35, TR34, RP33, PV32, VL31, LN30, NR29, RL28 , LF27, FD26, DK25, KI24, IR23, RQ22, QV21, VI20, IR19, ET36, TR35, RP34, PV33, VL32, LN31, NR30, RL29, LF28, FD27, DK26, KI25, IR24, RQ23, QV22, VI21 , IRK20, ET37, TR36, RP35, PV34, VL33, LN32, NR31, RL30, LF29, FD28, DK27, KI26, IR25, RQ24, QV23, VI22, IR21, ET38, TR37, RP36, PV35, VL34, LN33, NR32 , RL31, LF30, FD29, DK28, KI27, IR26, RQ25, QV24, VI23, IR22, ET39, TR38, RP37, PV36, VL35, LN34, NR33, RL32, LF31, FD30, DK29, KI28, IR27, RQ26, QV25 , VI24, IR23, ET40, TR39, R P38, PV37, VL36, LN35, NR34, RL33, LF32, FD31, DK30, KI29, IR28, RQ27, QV26, VI25, IRK24; and characterized in that it has the amino acid sequence of SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 3, SEQ ID NO: 4, SEQ ID NO: 5, SEQ ID NO: 6, SEQ ID NO: 7, SEQ ID NO: 8, SEQ ID NO: 9, SEQ ID NO: 10, SEQ ID NO: 11, SEQ ID NO: 12, SEQ ID NO: 13, SEQ ID NO: 14, SEQ ID NO: 15, SEQ ID NO: 16, SEQ ID NO: 17, SEQ ID NO: 18, SEQ ID NO: 19, SEQ ID NO: 20, SEQ ID NO: 21, SEQ ID NO: 22, SEQ ID NO: 23, SEQ ID NO: 24, SEQ ID NO: 25, SEQ ID NO: 26, SEQ ID NO: 27, SEQ ID NO: 28, SEQ ID NO: 29, SEQ ID NO: 30, SEQ ID NO: 31, SEQ ID NO: 32, SEQ ID NO: 33, SEQ ID NO: 34, SEQ ID NO: 35, SEQ ID NO: 36, SEQ ID NO: 37, SEQ ID NO: 38, SEQ ID NO: 39, SEQ ID NO: 40, SEQ ID NO: 41, SEQ ID NO: 42, SEQ ID NO: 43, SEQ ID NO: 44, SEQ ID NO: 45, SEQ ID NO: 46, SEQ ID NO: 47, SEQ ID NO: 48, SEQ ID NO: 49, SEQ ID NO: 50, SEQ ID NO: 51, SEQ ID NO: 52, SEQ ID NO: 53, SEQ ID NO: 54, SEQ ID NO: 55, SEQ ID NO: 56, SEQ ID NO: 57, SEQ ID NO: 58, SEQ ID NO: 59, SEQ ID NO: 60, SEQ ID NO: 61, SEQ ID NO: 62, SEQ ID NO: 6 3, SEQ ID NO: 64, SEQ ID NO: 65, SEQ ID NO: 66, SEQ ID NO: 67, SEQ ID NO: 68, SEQ ID NO: 69, SEQ ID NO: 70, SEQ ID NO: 71, SEQ ID NO: 72, SEQ ID NO: 73, SEQ ID NO: 74, SEQ ID NO: 75, SEQ ID NO: 76, SEQ ID NO: 77, SEQ ID NO: 78, SEQ ID NO: 79, SEQ ID NO: 80, SEQ ID NO: 81, SEQ ID NO: 82, SEQ ID NO: 83, SEQ ID NO: 84, SEQ ID NO: 85, SEQ ID NO: 86, SEQ ID NO: 87, SEQ ID NO: 88, SEQ ID NO: 89, SEQ ID NO: 90, SEQ ID NO: 91, SEQ ID NO: 92, SEQ ID NO: 93, SEQ ID NO: 94, SEQ ID NO: 95, SEQ ID NO: 96, SEQ ID NO: 97, SEQ ID NO: 98, SEQ ID NO: 99, SEQ ID NO: 100, SEQ ID NO: 101, SEQ ID NO: 102, SEQ ID NO: 103, SEQ ID NO: 104, SEQ ID NO: 105, SEQ ID NO: 106, SEQ ID NO: 107, SEQ ID NO: 108, SEQ ID NO: 109, SEQ ID NO: 110, SEQ ID NO: 111, SEQ ID NO: 112, SEQ ID NO: 113, SEQ ID NO: 114, SEQ ID NO: 115, SEQ ID NO: 116, SEQ ID NO: 117, SEQ ID NO: 118, SEQ ID NO: 119, SEQ ID NO: 120, SEQ ID NO: 121, SEQ ID NO: 122, SEQ ID NO: 123, SEQ ID NO: 124, SEQ ID NO: 125, SEQ ID NO: 126, SEQ ID NO: 127, SEQ ID NO: 128, SEQ ID NO: 129, SEQ ID NO: 130, SEQ ID NO: 131, SEQ ID NO: 132 , SEQ ID NO: 133, SEQ ID NO: 134, SEQ ID NO: 135, SEQ ID NO: 136, SEQ ID NO: 137, SEQ ID NO: 138, SEQ ID NO: 139, SEQ ID NO: 140, SEQ ID NO: 141, SEQ ID NO: 142, SEQ ID NO: 143, SEQ ID NO: 144, SEQ ID NO: 145, SEQ ID NO: 146, SEQ ID NO: 147, SEQ ID NO: 148, SEQ ID NO : 149, SEQ ID NO: 150, SEQ ID NO: 151, SEQ ID NO: 152, SEQ ID NO: 153, SEQ ID NO: 154, SEQ ID NO: 155, SEQ ID NO: 156, SEQ ID NO: 157 , SEQ ID NO: 158, SEQ ID NO: 159, SEQ ID NO: 160, SEQ ID NO: 161, SEQ ID NO: 162, respectively. 3. The polypeptide according to any one of claims 1 to 2, characterized in that the alpha-amino group of the N-terminal amino acid is acetylated and the alpha-carboxyl group of the C-terminal amino acid is amidated. 4. The use of the polypeptide according to any one of claims 1 to 3 to obtain a medicinal product for the prevention or treatment of bacterial infections of humans. 5. A medicament for the prevention or treatment of human bacterial infection, comprising the polypeptide according to any one of claims 1 to 3 and one or more pharmaceutically acceptable carriers, excipients or additional antibacterial agents. 6. The drug according to claim 5, composed, manufactured and intended for parenteral use. 7. The drug according to claim 5, compiled, manufactured and intended for local use on the mucous membrane of the affected area or tissue, for example, in the form of an irrigation liquid, ear drops, nose drops, aerosol, powder aerosol, gel, cream, lotion , suspension or mucoadhesive dosage form. 8. The use of a medicament according to any one of claims 5 to 7 for the prevention or treatment of infections of the respiratory tract or respiratory system, infections of the liver, spleen, eye, kidney, vagina, urethra, heart, infections or skin disorders, such as atopic dermatitis, chronic ulcer, diabetic foot, venous ulcer, acne, trauma or burn.

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