JP6042106B2 - Novel polypeptide containing methionine sulfoxide - Google Patents

Description

  The present invention relates to a novel polypeptide having angiogenesis-inducing activity and antibacterial activity, and an angiogenesis-inducing agent and / or antibacterial agent containing the same.

  Angiogenesis is effective in the treatment of various diseases or injuries such as burns, pressure ulcers, wounds, skin ulcers, leg ulcers, diabetic ulcers, occlusive arterial disease and occlusive arteriosclerosis. Moreover, in such diseases, since a bacterial infection induces a serious exacerbation of a disease state, an angiogenesis inducing agent having both antibacterial activity and angiogenesis inducing activity is required.

  Antibiotics are often used for bacterial infections. However, antibiotics are completely ineffective, especially in patients with weakened immunity, elderly people, and children due to the emergence of resistant bacteria. Examples have also been recognized, and new antibacterial agents other than antibiotics that do not produce resistant bacteria are also demanded.

  LL-37 is known as a polypeptide having angiogenesis-inducing activity and antibacterial activity. (Non-Patent Documents 1 and 2).

  Moreover, Nakagami et al. Invented a polypeptide having vascular endothelial growth activity and thus angiogenesis-inducing activity, and found that the peptide had angiogenesis-inducing activity higher than that of LL-37. Patent Document 1).

  Furthermore, Nakagami et al. Found a polypeptide AG30-5C consisting of 30 amino acid residues having higher angiogenesis-inducing activity than the peptide according to Patent Document 1, and applied for a patent (Patent Document 2).

  Patent Documents 1 and 2 also disclose that the polypeptide has antibacterial activity.

In addition, Proadrenomedullin NH ₂ -Terminal 20 peptide (PAMP) is known as a peptide that exhibits an angiogenesis-inducing action at a low dose. (Non-Patent Document 1)

WO2005 / 090564 WO2008 / 096816 WO2010 / 061915 WO2010 / 137594

Endocrinology, 2006, 147 (7), 3457-61.

  An object of the present invention is to provide a novel polypeptide having superior angiogenesis-inducing activity and antibacterial activity and a novel angiogenesis-inducing agent and / or antibacterial agent containing the same as an active ingredient.

  The present inventors have found a polypeptide having a higher vascular endothelial growth activity and, moreover, angiogenesis-inducing activity than the previous polypeptide. In addition, these polypeptides had higher antibacterial activity than conventional polypeptides.

As a result of diligent research, the present inventors have searched for the analogs of the polypeptides described in Patent Documents 1 and 2, and surprisingly, by substituting one or more methionine residues with methionine sulfoxide, The present invention was completed by finding that it has higher angiogenesis-inducing activity and antibacterial activity than polypeptides.

That is, the present invention more specifically comprises any of the following inventions.
(1) any one of the polypeptides represented by the following amino acid sequence or a pharmacologically acceptable salt thereof. (In the sequence, M (O) means methionine sulfoxide, and K * means D-lysine.)
M (O) LKLIFLHRLKRMRKRLK * RK (SEQ ID NO: 11)
MLKLIFLHRLKRM (O) RKRLK * RK (SEQ ID NO: 12)
M (O) LKLIFLHRLKRM (O) RKRLK * RK (SEQ ID NO: 13)
(2) The polypeptide of (1), which is a polypeptide represented by SEQ ID NO: 12, or a pharmacologically acceptable salt thereof , or a pharmacologically acceptable salt thereof.
(3) The polypeptide according to (1) or (2) , wherein one or more amino acid residues are D-amino acids, or a pharmaceutically acceptable salt thereof.
(4) The polypeptide according to any one of (1) to (3), wherein one or more lysine residues (K) are D-lysine, or a pharmaceutically acceptable salt thereof.
(5) The polypeptide according to any one of (1) to (4), wherein the C-terminus is an amide (—CONH ₂ ), or a pharmaceutically acceptable salt thereof.
(6) The polypeptide according to any one of (1) to (5), wherein the N-terminus is acetylated (CH ₃ CO-), or a pharmaceutically acceptable salt thereof.
(7) pharmaceutically acceptable salt is a hydrochloride or acetate salt, (1) a polypeptide or a pharmaceutically acceptable salt thereof, according to any one of - (6).
(8) An angiogenesis-inducing agent comprising the polypeptide according to any one of (1) to (7) or a pharmacologically acceptable salt thereof as an active ingredient.
(9) An antibacterial agent comprising the polypeptide according to any one of (1) to (8) or a pharmacologically acceptable salt thereof as an active ingredient

  According to the present invention, a polypeptide having excellent angiogenesis-inducing activity and antibacterial activity is provided. In addition, a novel angiogenesis inducer and / or antibacterial agent comprising this polypeptide as an active ingredient has been provided.

It is a chart figure of the MALDI-TOF / MS spectrum of the novel polypeptide represented by SEQ ID NO: 11. It is a chart figure of the MALDI-TOF / MS spectrum of the novel polypeptide represented by SEQ ID NO: 12. It is a chart figure of the MALDI-TOF / MS spectrum of the novel polypeptide represented by SEQ ID NO: 13. It is a figure which shows the result of having investigated the lumen formation in vitro of polypeptide of this invention. The area and length of the formed lumen are measured and expressed as a relative value at the time of peptide addition when the value of the negative control (Control) group is 100.

  In the present specification, the terms “polypeptide” and “peptide” are used synonymously.

  The amino acid sequence of polypeptide AG30-5C described in Patent Document 2 is shown in SEQ ID NO: 14. AG30-5C has antibacterial activity and angiogenesis-inducing activity, as specifically described in Patent Document 2 and the following examples. However, as specifically described in the following examples, the polypeptides according to the examples of the present invention had higher angiogenesis-inducing activity and antibacterial activity than AG30-5C.

  In addition, all the polypeptides that are active ingredients of the angiogenesis-inducing agent and / or antibacterial agent of the present invention are novel substances.

  In general, in a pharmaceutical comprising a polypeptide, a sugar chain or a polyethylene glycol (PEG) chain is added to the polypeptide or at least a part of the amino acids constituting the polypeptide in order to increase the stability of the polypeptide in vivo. A technique using a D-form amino acid is widely known and used. By adding sugar chains or PEG chains, or by using D-form amino acids as at least part of the amino acids that make up the polypeptide, it is less susceptible to degradation by peptidases in vivo, reducing the polypeptide in vivo by half. The period becomes longer. It is also well known that peptide stability is improved by N-terminal acetylation and / or C-terminal amidation of peptides. As long as the polypeptide of the present invention has antibacterial activity, it may be those subjected to these known modifications for in vivo stabilization, and the term “polypeptide” in the present specification and claims. Is used in the sense of including those that have been modified for in vivo stabilization, unless otherwise apparent from the context.

  Glycosylation to polypeptides is well known, for example, Sato M, Furuike T, Sadamoto R, Fujitani N, Nakahara T, Niikura K, Monde K, Kondo H, Nishimura S., "Glycoinsulins: dendritic sialyloligosaccharide-displaying insulins showing a prolonged blood-sugar-lowering activity. ", J Am Chem Soc. 2004 Nov 3; 126 (43): 14013-22 and Sato M, Sadamoto R, Niikura K, Monde K, Kondo H, Nishimura S," Site- specific introduction of sialic acid into insulin. ", Angew Chem Int Ed Engl. 2004 Mar 12; 43 (12): 1516-20. The sugar chain can be bound to the N-terminus, C-terminus, or an amino acid between them, but is preferably bound to the N-terminus or C-terminus in order not to inhibit or attenuate the activity of the polypeptide. The number of sugar chains to be added is preferably 1 or 2, and more preferably 1. The sugar chain is preferably monosaccharide to tetrasaccharide, more preferably disaccharide or trisaccharide. The sugar chain can be bound to a free amino group or carboxyl group of the polypeptide directly or via a spacer structure such as a methylene chain having about 1 to 10 carbon atoms.

  Addition of PEG chains to polypeptides is also well known, e.g. Ulbricht K, Bucha E, Poschel KA, Stein G, Wolf G, Nowak G., "The use of PEG-Hirudin in chronic hemodialysis monitored by the Ecarin Clotting Time: influence on clotting of the extracorporeal system and hemostatic parameters. ", Clin Nephrol. 2006 Mar; 65 (3): 180-90. and Dharap SS, Wang Y, Chandna P, Khandare JJ, Qiu B, Gunaseelan S, Sinko PJ, Stein S, Farmanfarmaian A, Minko T., "Tumor-specific targeting of an anticancer drug delivery system by LHRH peptide.", Proc Natl Acad Sci USA. 2005 Sep 6; 102 (36): 12962-7. The PEG chain can be attached to the N-terminus, C-terminus, or the amino acid between them, and usually one or two PEG chains are attached to a free amino group or carboxyl group on the polypeptide. The molecular weight of the PEG chain is not particularly limited, but is usually about 3000 to 7000, preferably about 5000.

  A method in which at least a part of amino acids constituting a polypeptide is in D form is also well known. For example, Brenneman DE, Spong CY, Hauser JM, Abebe D, Pinhasov A, Golian T, Gozes I., “Protective peptides that are orally active and mechanistically nonchiral. ", J Pharmacol Exp Ther. 2004 Jun; 309 (3): 1190-7, Wilkemeyer MF, Chen SY, Menkari CE, Sulik KK, Charness ME.," Ethanol antagonist peptides: structural specificity without stereospecificity ., J Pharmacol Exp Ther. 2004 Jun; 309 (3): 1183-9. A part of the amino acids constituting the polypeptide may be D-form, or all of the constituent amino acids may be D-form amino acids as long as the activity of the polypeptide is not inhibited or attenuated. More specifically, for example, SEQ ID NOs: 11 to 13 can be mentioned.

  Here, as pharmacologically acceptable salts according to the present invention, for example, salts with inorganic acids such as hydrochloric acid, hydrobromic acid, sulfuric acid, nitric acid, phosphoric acid, acetic acid, trifluoroacetic acid, lactic acid, tartaric acid, Examples thereof include salts with organic acids such as oxalic acid, maleic acid, methanesulfonic acid, benzenesulfonic acid, and p-toluenesulfonic acid, among which hydrochloride, acetate, and trifluoroacetate are more preferable.

  Furthermore, the polypeptide of the present invention is not limited to the above-described one as long as it has substantially the same effect, and includes, for example, an amino acid sequence in which one or more amino acids are substituted, deleted, and / or added. Polypeptides are also included.

  The polypeptide which is an active ingredient of the angiogenesis inducer of the present invention can be easily produced by a conventional method such as chemical synthesis using a commercially available peptide synthesizer. The stabilization modification can also be easily performed by a known method as described in each of the above documents.

  Since the polypeptide of the present invention has high angiogenesis inducing activity and antibacterial activity, it can be used as an angiogenesis inducing agent and / or an antibacterial agent. The method of use as an angiogenesis inducing agent and / or antibacterial agent is the same as that of known polypeptide angiogenesis inducing agents or antibacterial agents, and includes solutions, emulsions, suspensions, powders, powders, granules, gels, It can be administered as an ointment or transdermal patch, particularly preferably as a solution, powder or transdermal patch. The solution is preferably a buffer solution, particularly preferably a solution dissolved in an aqueous medium such as physiological saline, phosphate buffer, citrate buffer or the like. The concentration of the polypeptide in the solution is not particularly limited, but is usually about 0.01 mg / mL to 100 mg / mL, preferably about 0.1 mg / mL to 50 mg / mL, particularly preferably about 1 mg / mL to 10 mg / mL. is there. The route of administration is usually local administration such as application, spraying, contact (sticking, covering) or injection to a site requiring angiogenesis and / or sterilization. The dosage is appropriately selected according to the symptoms and the size of the affected area, but is usually about 0.01 mg to 100 mg, preferably about 0.05 mg to 50 mg, particularly preferably about 0.1 mg to 10 mg as the polypeptide amount. However, of course, it is not limited to these ranges. As the pharmaceutically acceptable carrier used in formulating the angiogenesis inducer and / or antibacterial agent of the present invention, a carrier commonly used in the pharmaceutical field is used in addition to the above-mentioned aqueous medium. For example, in the case of an external preparation such as an ointment, hydrocarbons (hydrophilic petrolatum, white petrolatum, purified lanolin, liquid paraffin, etc.), zinc oxide, higher fatty acids and esters thereof (adipic acid, myristic acid, Palmitic acid, stearic acid, oleic acid, adipic acid ester, myristic acid ester, palmitic acid ester, diethyl sebacate, hexyl laurate, cetyl isooctanoate, etc.) wax (whale wax, beeswax, ceresin etc.), higher alcohol ( Cetanol, stearyl alcohol, cetostearyl alcohol, etc.). In the case of a solution, for example, water, physiological saline, phosphate buffered saline and the like can be mentioned, and in the case of an oral preparation, lactose, starch and the like can be mentioned. In addition, various pharmaceutical additives such as an emulsifier, a surfactant, a tonicity agent, and a pH adjuster can be blended as necessary. These pharmaceutically acceptable carriers and pharmaceutical additives are well known and widely used in the pharmaceutical field.

  Examples of specific diseases and disorders when administered to living bodies include ulcers caused by burns, pressure ulcers, wounds, skin ulcers, lower limb ulcers, diabetic ulcers, occlusive arterial disease, obstructive arteriosclerosis, etc. However, it is not limited to these. The angiogenesis-inducing agent of the present invention can be used as a preventive, ameliorating or therapeutic agent for these diseases or disorders. The polypeptide of the present invention has a high angiogenesis-inducing activity and also has an antibacterial activity. Therefore, the polypeptide of the present invention is particularly suitable as a preventive, ameliorating or therapeutic agent for a disease or disorder desired to have an antibacterial activity. Examples of such diseases or disorders include burns, pressure ulcers, wounds, skin ulcers, lower limb ulcers, diabetic ulcers, etc. among the above diseases or disorders.

  The angiogenesis-inducing agent of the present invention can be used alone, or can be used in combination with other antibacterial agents or antibiotics when further antibacterial properties are desired. Examples of these antibacterial agents or antibiotics include cephem series, carbapenem series, aminoglycoside series, new quinolone series, β-lactam series, penicillin series and glycopeptide series antibiotics, and more specifically, ceftazidime. , Meropenem, tobramycin, ciprofloxacin, methicillin, ampicillin, vancomycin and the like, but are not limited thereto.

  Hereinafter, the present invention will be described more specifically based on examples. However, the present invention is not limited to the following examples.

1. Synthesis of polypeptides Following the methods described in the literature Solid Phase Peptide Synthesis, Pierce (1984), Fmoc solid synthesis: a practical approach, Oxford University Press (2000) and 5th edition, Experimental Chemistry Course 16, Synthesis of Organic Compounds IV, etc. Using an automatic solid phase synthesizer, a protected peptide resin was synthesized by the Fmoc method. Trifluoroacetic acid (TFA) and scavenger (a mixture of thioaniol, ethanedithiol, phenol, triisopropylsilane, water, etc.) were added to the obtained protected peptide resin, and the crude peptide was obtained by cutting out from the resin and deprotecting it. . The crude peptide was purified using a reverse phase HPLC column (ODS) by gradient elution with a 0.1% TFA-H ₂ O / CH ₃ CN system. Fractions containing the desired product were collected and lyophilized to obtain the desired peptide.
The sequence of the peptide is shown below. (In the sequence, M (O) means methionine sulfoxide, and K * means D-lysine.)
(SEQ ID NO: 1)
M (O) LKLIFLHRLKRMRKRLKRK
(SEQ ID NO: 2)
MLKLIFLHRLKRM (O) RKRLKRK
(SEQ ID NO: 3)
M (O) LKLIFLHRLKRM (O) RKRLKRK
(SEQ ID NO: 4)
ELRFLHRLKRM (O) RKRLKRKLR
(SEQ ID NO: 5)
KLIFLHRLKRM (O) RKRLKRKLR
(SEQ ID NO: 6)
KRM (O) RKRLKRKLRLWHRKRYK
(SEQ ID NO: 7)
M (O) RKRLKRKLRLWHRKRYK
(SEQ ID NO: 8)
M (O) LKLIFLHRLKRMRKRLKRKLRFWHRKRYK
(SEQ ID NO: 9)
MLKLIFLHRLKRM (O) RKRLKRKLRFWHRKRYK
(SEQ ID NO: 10)
M (O) LKLIFLHRLKRM (O) RKRLKRKLRFWHRKRYK

(SEQ ID NO: 11)
M (O) LKLIFLHRLKRMRKRLK * RK
(SEQ ID NO: 12)
MLKLIFLHRLKRM (O) RKRLK * RK
(SEQ ID NO: 13)
M (O) LKLIFLHRLKRM (O) RKRLK * RK

(SEQ ID NO: 14) AG30-5C
MLSLIFLHRLKSMRKRLDRKLRLWHRKNYP
(SEQ ID NO: 15) Comparative control
MLKLIFLHRLKRMRKRLK * RK

2. Analysis of polypeptide using MALDI-TOF / MS The sequence of the synthesized polypeptide represented by SEQ ID NOs: 11 to 13 was confirmed by the analysis result using MALDI-TOF / MS. 1 μL of a matrix solution (α-Cyano 4-Hydroxy Cinnamic Acid) was added to 1 μL of a polypeptide 0.1% TFA / 50% acetonitrile solution having a final concentration of 100 μg / mL to obtain a measurement sample for MALDI. A measurement sample for MALDI (0.4 μL) was applied to a MALDI target plate and dried. Measurement was performed using MALDI-TOF / MS.

MALDI-TOF / MS conditions:
Laser intensity: 2100
Number of shots: 1000

Results Table 1 shows the theoretical and actual MALDI-TOF / MS values of each polypeptide. Moreover, the MALDI-TOF / MS spectrum (monoisotopic) of each polypeptide was shown in FIGS. The detected m / z of each polypeptide agreed with the respective theoretical value, and the sequence of the synthesized polypeptide could be confirmed.

The results are shown in Table 1.
[Table 1]
───────────────────────
SEQ ID NO: 11 SEQ ID NO: 12 SEQ ID NO: 13
───────────────────────
Theoretical 2,681.4 2,681.4 2,697.4
Actual value 2,681.5 2,681.5 2,697.5
───────────────────────

3. Angiogenesis-inducing activity of polypeptides Polypeptide formation in vitro was examined using angiogenesis kits (Angiogenesis Kit, KZ-1000, KURABO, trade name). VEGF-A (included in the kit) was used as a positive control, and an unstimulated group without peptide was used as a negative control. Each polypeptide concentration was adjusted to 10, 2.5, 0.5 mg / mL in angiogenesis medium. Cells were added to an angiogenesis medium containing the polypeptide and cultured under conditions of 37 ° C. and 5% CO ₂ . The medium was replaced with one containing the same polypeptide on days 4, 7 and 9 of culture. On day 11 after the start of culture, the medium was removed, and staining was performed using a luminal staining kit (for CD31 antibody staining) (KURABO, trade name) according to the following procedure.
After removing the medium from each well, the cells were washed with 1 mL of PBS (−), and 1 mL of 70% ethanol was added to fix the cells. CD31-stained primary antibody (mouse anti-human CD31 antibody) was diluted 4000 times with a blocking solution (Dulbecco phosphate buffer solution (PBS (−)) containing 1% BSA). 0.5 mL of this primary antibody solution was added to each well and incubated at 37 ° C. for 60 minutes. After the incubation, each well was washed 3 times with 1 mL of blocking solution.
Next, 0.5 mL of a secondary antibody solution (goat anti-mouse IgG alkaline phosphatase complex) diluted 500 times with blocking solution was added to each well, incubated at 37 ° C. for 60 minutes, and then washed 3 times with 1 mL of distilled water. . Meanwhile, two BCIP / NBT tablets were dissolved in distilled water and filtered through a filter with a pore size of 0.2 mm to prepare a substrate solution. 0.5 mL of the adjusted BCIP / NBT substrate solution was added to each well and incubated at 37 ° C. until the lumen became deep purple (usually 5-10 minutes). After the incubation, each well was washed 3 times with 1 mL of distilled water. After washing, the washing solution was removed by suction and left to stand to dry naturally. After drying, photographs of each well were taken under a microscope. Using angiogenesis quantification software (KURABO, trade name), the area and length of the lumen formed from each image data were measured, and evaluated by the relative value when the peptide was added to the negative control.

  The results are shown in Table 2.

[Table 2]
─────────────────────────────────
Polypeptide concentration Area and length of lumen
(μg / mL) (Rate with negative control as 100)
─────────────────────────────────
Control (SEQ ID NO: 15) 0.5 122.8
2.5 191.4
The peptide of the present invention (SEQ ID NO: 11) 0.5 162.3
2.5 176.7
The peptide of the present invention (SEQ ID NO: 12) 0.5 169.6
2.5 246.6
The peptide of the present invention (SEQ ID NO: 13) 0.5 170.2
2.5 208.4
─────────────────────────────────

  As shown in Table 2, all of the polypeptides represented by SEQ ID NOs: 11, 12, and 13 of the present invention had angiogenesis-inducing activity.

4). Antibacterial Activity of Polypeptide The antibacterial activity was evaluated according to the procedure of the International Committee for Clinical Laboratory Standard Institute (CLSIM7-A7). That is, using a microtiter plate or a test tube, peptides that inhibit the growth of bacteria at a peptide concentration of 32 μg / mL were screened. As strains, Staphylococcus aureus ATCC29213 (S. aureus ATCC29213) Pseudomonas aeruginosa ATCC27853 (P. aeniginosa ATCC27853) and Escherichia coli ATCC25922 (Escherichia coli ATCC25922) were used. The bacteria were cultured in the medium for 4-6 hours and then diluted with Mueller-Hinton broth (MHB) according to McFarland # 0.5. Each strain was added to a concentration of about 1 × 10 ⁵ CFU / mL (final concentration). Each peptide was adjusted to an arbitrary concentration, and serial dilution was performed therefrom. The bacterial solution was added to a microplate or test tube into which peptides at each concentration step were dispensed. The one without peptide was used as a negative control, and gentamicin was used as a positive control. The plate was incubated at 37 ° C. for 20 minutes, and the peptide concentration that inhibited the growth of the bacteria was defined as the minimum growth inhibitory concentration (MIC) (μg / mL).

  The results are shown in Table 3.

[Table 3]
────────────────────────────────
Polypeptide MIC (μg / mL)
E. coli Pseudomonas aeruginosa Staphylococcus aureus ─────────────────────────────────
Control (SEQ ID NO: 15) 32 8/16 8/16
The peptide of the present invention (SEQ ID NO: 11) 16/32 16 32
The peptide of the present invention (SEQ ID NO: 12) 16 16 8/16
The peptide of the present invention (SEQ ID NO: 13) 16/32 32 32
────────────────────────────────

  As shown in Table 3, all the polypeptides represented by SEQ ID NOs: 11, 12, and 13 of the present invention had antibacterial activity.

5. Effects of polypeptides on skin fibroblast proliferation.
The human skin fibroblast proliferation activity of the peptides was examined using Cell Counting Kit (WST-1) (trade name, Dojindo). As a negative control, an unstimulated group without peptide was used. Normal human skin fibroblasts (NHDF) were seeded in a 96-well plate (0.5 × 10 ⁴ cells / well / 100 μL, 1% FBS). About 3 hours after seeding the cells, 100 μL of candidate peptide or bFGF (positive control) was added. Only 100 μL of Medium was added to the unstimulated group. It was allowed to stand for about 48 hours in a CO ₂ incubator. After 48 hours, 20 μl of WST-1 reagent was added to each well and allowed to stand in a CO ₂ incubator for about 2 hours. Absorbance at wavelengths of 450 nm and 620 nm was measured with Wallac 1420 ARVOsx (Perkin Elmer program: WST-1). OD450-OD620 of each measured value was obtained. NetO.D.450 was obtained by subtracting the average value of OD450-OD620 of empty wells not containing cells from the value of OD450-OD620 of measurement wells. Cell proliferation activity was evaluated by the ratio at the time of peptide addition to unstimulated NetO.D.450.

  The results are shown in Table 4.

[Table 4]
─────────────────────────────────────
Polypeptide NHDF growth (μg / mL)
1 3 10 30 100
─────────────────────────────────────
Control (SEQ ID NO: 15) 119.08 130.64 146.78 116.27 -1.93
Peptide of the present invention (SEQ ID NO: 11) 112.42 118.97 120.73 131.68 130.26
Peptide of the present invention (SEQ ID NO: 12) 114.25 120.13 137.55 151.84 -1.37
Peptide of the present invention (SEQ ID NO: 13) 115.10 115.14 122.21 137.66 140.14
─────────────────────────────────────

  As shown in Table 4, all the polypeptides represented by SEQ ID NOs: 11, 12, and 13 of the present invention had human skin fibroblast proliferation activity.

Claims (9)

Any one of the polypeptides represented by the following amino acid sequence or a pharmacologically acceptable salt thereof. (In the sequence, M (O) means methionine sulfoxide, and K * means D-lysine.)
M (O) LKLIFLHRLKRMRKRLK * RK (SEQ ID NO: 11)
MLKLIFLHRLKRM (O) RKRLK * RK (SEQ ID NO: 12)
M (O) LKLIFLHRLKRM (O) RKRLK * RK (SEQ ID NO: 13) The polypeptide according to claim 1, which is a polypeptide represented by SEQ ID NO: 12, or a pharmacologically acceptable salt thereof , or a pharmacologically acceptable salt thereof. The polypeptide according to claim 1 or 2, wherein one or more amino acid residues are D-amino acids, or a pharmaceutically acceptable salt thereof. The polypeptide according to any one of claims 1 to 3, wherein the one or more lysine residues (K) are D-lysine, or a pharmaceutically acceptable salt thereof. C-terminal is an amide (-CONH _2), a polypeptide according to any one of claims 1-4 or a pharmaceutically acceptable salt thereof. N-terminal is acetylated (CH ₃ CO-) of, a polypeptide according to any one of claims 1-5 or a pharmaceutically acceptable salt thereof. Pharmacologically acceptable salt is a hydrochloride salt or acetate, polypeptide according to any one of claims 1 to 6 or a pharmacologically acceptable salt thereof. An angiogenesis inducer comprising the polypeptide according to any one of claims 1 to 7 or a pharmacologically acceptable salt thereof as an active ingredient. An antibacterial agent comprising the polypeptide according to any one of claims 1 to 8 or a pharmacologically acceptable salt thereof as an active ingredient.

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