JP2011195451A - Peptide - Google Patents

Abstract

The present invention provides a peptide that can be used to prevent infection in the medical and environmental health fields, and to treat or reduce the incidence of infection, an antibacterial composition containing the peptide, and an antibacterial drug.
The peptide has the formula [I]: Ile-Leu-Arg-X1-X2-X3-X4-X5-X6-Arg-Arg-X7- (X8) n (wherein X1 is Trp or thiotryptophan) (Ws), X2 is Pro or Ala, X3 is Trp or Ws, X4 is Trp, Ws or Lys, X5 is Pro or Ala, X6 is Trp or Ws, X7 is Lys or Trp, X8 is Ala-X9-Ala- Ala (X9 is Ala or Arg), n is 0 or 1, and each amino acid is L or D. However, X1, X3, X4 and X6 are Trp, X2 and X5 are Pro, X7 Is 1 and when the amino acid is L-form, n is 1).
[Selection figure] None

Description

  The present invention relates to a peptide that can be used particularly for various medical uses and environmental hygiene uses. More particularly, the present invention relates to peptides that can be used to prevent infections in various medical and environmental health fields, and to treat or reduce the incidence of infectious diseases, in particular antibacterial peptides and antibacterial properties containing them. The present invention relates to a composition and an antibacterial medicine containing them.

  Numerous medicines such as antibiotics effective against infectious diseases caused by bacteria, fungi, viruses and the like have been developed and used in the medical field and the environmental health field. Despite the fact that antibiotics that are effective against various bacteria are used, not only in developing countries but also in developed countries, many people receive various bacteria, fungi, viruses, etc. every year. Infected with the infectious disease that caused it, and died of the infection.

  Also, in the field of medical and environmental health fields, various infectious disease prevention measures have been taken so far, and infection control efforts have been made for the purpose of preventing infection epidemics in medical facilities. ing. However, in such a situation, despite the fact that thorough infection prevention measures and infection control are being taken, it is a fact that we are facing the challenges of infection control and infection management due to nosocomial infections and opportunistic infections. .

  Furthermore, various medical devices and tools are frequently used in medical sites. At present, many implantable or long-term indwelling medical devices and tools such as suture tools, orthopedic devices, stents, catheters, guidewires, shunts, prosthetic heart valves, artificial joints, heart pacemakers, etc. are used. ing. However, it is also true that these medical devices and tools are one of the main causes of infectious diseases.

  Problems such as those mentioned above in the field of medical and environmental health such as bacteria, fungi, which cause infections due to frequent use of drugs such as antibiotics and heavy use of many kinds of drugs, etc. One of the causes is that the effectiveness of such drugs is reduced or the effect is lost due to acquisition of resistance to the drug by microorganisms such as viruses. Furthermore, the emergence of so-called multi-drug resistant bacteria in which a single microorganism is resistant to several types of drugs at the same time makes infection control more difficult.

  Therefore, there has been a demand for a different type of drug from drugs that have already developed resistant bacteria, such as conventional antibiotics. As one of the drugs that have been developed to meet such demands, so-called antibacterial peptides are attracting attention as substances that complement or substitute conventional antibiotics.

  An antibacterial peptide is one of the biological defense mechanisms originally provided for living organisms to protect themselves against external microorganisms. Since this antibacterial peptide is produced by living organisms, it has no or very little side effects and inhibitory effects on the living body, and has a broad antibacterial spectrum against bacteria, fungi, etc. .

  Such antibacterial peptides include, for example, naturally-occurring antibacterial peptides such as bactenecin, defensin, indolicidin, cinhistatin, lactoferrin and its degradation products lactofurine, magainin, sacropine, melittin, and maculatin, and synthetic antibacterial peptides such as histatin derivatives. Etc. are known (see, for example, Non-Patent Document 1 and Patent Document 1-10).

Among these antibacterial peptides, indolicidin is a peptide exhibiting a broad spectrum antibacterial activity consisting of 13 amino acids rich in tryptophan (tryptophan rich) isolated from bovine neutrophils and amidated at the carboxy terminus in its natural form. (For example, see Non-Patent Document 2 and Patent Document 4). However, since indolicidin itself has toxicity, studies on indolicidin analogues having lower toxicity and exhibiting broad spectrum antibacterial activity have been made (see, for example, Patent Documents 3-6). Of these peptides, Ominagan pentahydrochloride, a synthetic cationic antibacterial peptide that is one of indolicidin analogs, is currently being developed as a local infection preventive for catheters (for example, Non-Patent Document 2, 3 and 4).
FEBS Lett., 449, 105-110, 1999 Selsted, et al .; Biol. Chem. 267: 4292, 1992 Sadar, HS, et al .; Antimicrob. Agents Chemother. 2004 August, 48 (8): 3112-3118 Melo, MN, et al .; Recent Patents on Anti-Infective Drug Discovery, Volume 1, Number 2, June 2006, pp. 201-207 (7), published by Bentham Science Publishers Japanese National Patent Publication No. 8-508165 JP-T 9-509175 JP 2001-172297 A JP-T-2001-500477 WO-A1-98007745 JP 2002-515403 A JP 2002-179698 A JP 2003-52365 A JP 2005-154338 A U.S. Pat. No. 4,543,252

  Therefore, as a result of diligent research and examination, the present inventors further prepared a synthetic peptide of an analog of indolicidin, which is a tryptophan-rich (tryptophan-rich) peptide among the existing antibacterial peptides, and the bacteria of the synthetic peptide As a result of investigating antibacterial and antiviral activities against fungi and viruses, these peptides have antibacterial and antiviral activities effective against bacteria, fungi, viruses, etc. that are higher than or equivalent to omiganane. In addition, the present invention has been completed by finding that it has effective growth-inhibiting activity particularly against Gram-positive bacteria that are also the cause of nosocomial infections.

  Accordingly, an object of the present invention is to provide a novel peptide having an antibacterial and / or antiviral activity against bacteria, fungi and / or viruses, a derivative thereof and a salt thereof.

  More specifically, the present invention is a peptide that is an analog of indolicidin, which is a tryptophan-rich (tryptophan-rich) peptide, and has high antibacterial activity, antiviral activity, etc. against bacteria, fungi, viruses, etc. Another object of the present invention is to provide a peptide having effective growth inhibitory activity against Gram-positive bacteria having resistance to antibiotics, derivatives thereof, and salts thereof.

  In the present specification, even if it is simply described as “peptide”, unless otherwise specified, it is understood that the term “peptide” is used in the meaning including derivatives and salts of the peptide. be able to.

  The present invention contains at least one antibacterial composition comprising the above peptide and an antibiotic, antifungal agent, antiviral agent and / or antiparasitic agent, and the above peptide and / or the above antibacterial composition. It aims to provide antibacterial medicine.

In order to achieve the above object, the present invention provides a compound represented by the general formula [I]:
Ile-Leu-Arg-X1-X2-X3-X4-X5-X6-Arg-Arg-X7- (X8) n [I] (SEQ ID NO: 1)
(Where X1 is tryptophan (Trp) or chemical formula [II]:

で表されるチオトリプトファン (Ws) を意味し；
X2 はプロリン (Pro) またはアラニン (Ala) を意味し；
X3 はトリプトファン (Trp) またはチオトリプトファン (Ws) を意味し；
X4 はトリプトファン (Trp)、チオトリプトファン (Ws) またはリジン (Lys) を意味し；
X5 はプロリン (Pro) またはアラニン (Ala) を意味し；
X6 はトリプトファン (Trp) またはチオトリプトファン (Ws) を意味し；
X7 はリジン (Lys) またはトリプトファン(Trp) を意味し；
X8 は Ala - X9 - Ala - Ala（式中、X9 は Ala または Arg を意味する）（配列番号２）を意味し；
n は０または１を意味し;および
それぞれのアミノ酸が L− 体または D− 体である）を意味し；
ただし、X1、X3、X4 および X6 がそれぞれトリプトファン (Trp) を意味し；X2 および X5 がそれぞれプロリン(Pro) を意味し；X7 がリジン (Lys) を意味し、かつ、それぞれのアミノ酸が L− 体である場合は、n は１を意味する）
で表されるペプチドもしくはその誘導体またはそれらの塩を提供する。

Thiotryptophan (Ws) represented by
X2 means proline (Pro) or alanine (Ala);
X3 means tryptophan (Trp) or thiotryptophan (Ws);
X4 means tryptophan (Trp), thiotryptophan (Ws) or lysine (Lys);
X5 means proline (Pro) or alanine (Ala);
X6 means tryptophan (Trp) or thiotryptophan (Ws);
X7 means lysine (Lys) or tryptophan (Trp);
X8 means Ala-X9-Ala-Ala (where X9 means Ala or Arg) (SEQ ID NO: 2);
n represents 0 or 1; and each amino acid is L-form or D-form);
Where X1, X3, X4 and X6 each represent tryptophan (Trp); X2 and X5 each represent proline (Pro); X7 represents lysine (Lys) and each amino acid is L- If it is a field, n means 1)
Or a derivative thereof or a salt thereof.

The present invention provides a peptide having any amino sequence represented by the following formulas [III] to [VIII], a derivative thereof, or a salt thereof.
Ile-Leu-Arg-Ws-Pro-Ws-Ws-Pro-Ws-Arg-Arg-Lys [III] (SEQ ID NO: 3)
Ile-Leu-Arg-Trp-Pro-Trp-Trp-Pro-Trp-Arg-Arg-Lys-Ala-Ala-Ala-Ala
[IV] (SEQ ID NO: 4)
Ile-Leu-Arg-Trp-Ala-Trp-Lys-Ala-Trp-Arg-Arg-Trp-Ala-Ala-Ala-Ala
[V] (SEQ ID NO: 5)
Ile-Leu-Arg-Ws-Pro-Ws-Ws-Pro-Ws-Arg-Arg-Lys-Ala-Ala-Ala-Ala
[VI] (SEQ ID NO: 6)
Ile-Leu-Arg-Trp-Ala-Trp-Lys-Ala-Trp-Arg-Arg-Trp-Ala-Arg-Ala-Ala
[VII] (SEQ ID NO: 7)
Ile-Leu-Arg-Trp-Pro-Trp-Trp-Pro-Trp-Arg-Arg-Lys (all in D-form)
[VIII] (SEQ ID NO: 8)
(Wherein Ws has the same meaning as above).

  The present invention relates to a peptide comprising an amino acid sequence of 12 to 40, preferably 12 to 30, more preferably 12 to 20, particularly preferably 12 to 16, including the amino acid sequence of the peptide, or Derivatives or salts thereof are provided.

  In a more preferred embodiment of the present invention, the peptide has an antibacterial activity, antiviral activity against bacteria, fungi and / or viruses, or a growth inhibitory activity against gram-positive bacteria, or a combination thereof. I will provide a.

  As another form thereof, the present invention provides an antibacterial composition comprising the above peptide and an antibiotic, antifungal agent, antiviral agent, and / or antiparasitic agent.

  The present invention provides, as another form thereof, an antibacterial medicament containing at least one of the peptide and / or the antibacterial composition.

  Since the peptide according to the present invention has antibacterial activity, antiviral activity against bacteria, fungi, and / or viruses, or particularly anti-growth activity against Gram-positive bacteria, or a combination thereof, bacteria, fungi and It is extremely effective against infections caused by viruses, etc., and is effectively applied to the prevention and treatment of various infectious diseases by using in combination with antibiotics, antifungal agents, antiviral agents and / or antiparasitic agents. can do. Furthermore, since the peptide of the present invention is a different type of antibacterial peptide from conventional antibiotics, it has the great advantage of being effective against bacteria, fungi, etc. that are resistant to conventional antibiotics. is doing.

  The present invention is extremely effective for infection by bacteria, fungi and / or viruses having antibacterial and / or antiviral activity against bacteria, fungi and / or viruses, or, in particular, anti-growth activity against gram-positive bacteria. Antibacterial peptides containing antibacterial peptides that can be effectively applied to the prevention and treatment of various infectious diseases when used in combination with antibiotics, antifungal agents, antiviral agents, and / or antiparasitic agents A composition; and an antibacterial medicament comprising the antibacterial peptide or the antibacterial composition. Furthermore, since the peptide of the present invention is a different type of antibacterial peptide from conventional antibiotics, it has the great advantage of being effective against bacteria, fungi, etc. that are resistant to conventional antibiotics. is doing.

で表されるチオトリプトファン (Ws) を意味し；
X2 はプロリン (Pro) またはアラニン (Ala) を意味し；
X3 はトリプトファン (Trp) またはチオトリプトファン (Ws) を意味し；
X4 はトリプトファン (Trp)、チオトリプトファン (Ws) またはリジン (Lys) を意味し；
X5 はプロリン (Pro) またはアラニン (Ala) を意味し；
X6 はトリプトファン (Trp) またはチオトリプトファン (Ws) を意味し；
X7 はリジン (Lys) またはトリプトファン(Trp) を意味し；
X8 は Ala - X9 - Ala - Ala （式中、X9 は Ala または Arg を意味する）（配列番号２）を意味し；
n は０または１を意味し;および
それぞれのアミノ酸が L− 体または D− 体である）を意味し；
ただし、X1、X3、X4 および X6 がそれぞれトリプトファン (Trp) を意味し；X2 および X5 がそれぞれプロリン(Pro) を意味し；X7 がリジン (Lys) を意味し、かつ、それぞれのアミノ酸が L− 体である場合は、n は１を意味する）
で表されるペプチドまたはその誘導体もしくはその塩からなるペプチドである。 The peptide according to the present invention has the general formula [I]:
Ile-Leu-Arg-X1-X2-X3-X4-X5-X6-Arg-Arg-X7- (X8) n [I] (SEQ ID NO: 1)
(Where X1 is tryptophan (Trp) or chemical formula [II]:

Thiotryptophan (Ws) represented by
X2 means proline (Pro) or alanine (Ala);
X3 means tryptophan (Trp) or thiotryptophan (Ws);
X4 means tryptophan (Trp), thiotryptophan (Ws) or lysine (Lys);
X5 means proline (Pro) or alanine (Ala);
X6 means tryptophan (Trp) or thiotryptophan (Ws);
X7 means lysine (Lys) or tryptophan (Trp);
X8 means Ala-X9-Ala-Ala (where X9 means Ala or Arg) (SEQ ID NO: 2);
n represents 0 or 1; and each amino acid is L-form or D-form);
Where X1, X3, X4 and X6 each represent tryptophan (Trp); X2 and X5 each represent proline (Pro); X7 represents lysine (Lys) and each amino acid is L- If it is a field, n means 1)
Or a derivative or a salt thereof.

More specifically, the peptide of the present invention comprises a peptide having any amino sequence represented by the following formulas [III] to [VIII], a derivative thereof or a salt thereof.
Ile-Leu-Arg-Ws-Pro-Ws-Ws-Pro-Ws-Arg-Arg-Lys [III] (SEQ ID NO: 3)
Ile-Leu-Arg-Trp-Pro-Trp-Trp-Pro-Trp-Arg-Arg-Lys-Ala-Ala-Ala-Ala
[IV] (SEQ ID NO: 4)
Ile-Leu-Arg-Trp-Ala-Trp-Lys-Ala-Trp-Arg-Arg-Trp-Ala-Ala-Ala-Ala
[V] (SEQ ID NO: 5)
Ile-Leu-Arg-Ws-Pro-Ws-Ws-Pro-Ws-Arg-Arg-Lys-Ala-Ala-Ala-Ala
[VI] (SEQ ID NO: 6)
Ile-Leu-Arg-Trp-Ala-Trp-Lys-Ala-Trp-Arg-Arg-Trp-Ala-Arg-Ala-Ala
[VII] (SEQ ID NO: 7)
Ile-Leu-Arg-Trp-Pro-Trp-Trp-Pro-Trp-Arg-Arg-Lys (all in D-form)
[VIII] (SEQ ID NO: 8)
(Wherein Ws has the same meaning as above).

  In the peptide according to the present invention, as described above, thiotryptophan [II] represented by Ws is an unnatural amino acid in which the nitrogen atom of the indole ring of tryptophan is substituted with a sulfur atom.

  The peptide of the present invention has 12 to 40 amino acids, preferably 12 to 30, more preferably 12 to 20, and particularly preferably 12 to 16 amino acids in the amino sequence. Good. As long as the peptide of the present invention has an amino acid sequence consisting of 12 to 16 amino acids represented by the above general formula, and has antimicrobial activity such as antibacterial activity and antiviral activity which are the objects of the present invention, The types of other amino acids are not particularly limited, and can be selected from any amino acid.

  The peptide according to the present invention can be generally produced according to a peptide chemical synthesis method or the like commonly used in the art. The peptide chemical synthesis method includes a solid phase synthesis method and a liquid phase synthesis method (for example, a solid phase synthesis method and a liquid phase synthesis method; Nobuo Izumiya, Tetsuo Kato, Toshihiko Aoyagi, Michinori Waki , “Basics and Experiments of Peptide Synthesis”, 1985, Maruzen Co., Ltd.), currently, solid-phase synthesis is commonly used. Further, a commercially available peptide synthesizer (for example, an automatic peptide synthesizer (PTI-PS3: manufactured by Aroka Co., Ltd .; PSSM-8: manufactured by Shimadzu Corporation)) may be used.

  In general, the general method of peptide solid phase synthesis is to fix the amino acid carboxyl terminal to an insoluble solid resin, and then condense the amino acids one by one by active esterification etc. to extend the peptide chain. And then synthesizing the target peptide by desorption from the resin.

  More specifically, in the peptide of the present invention, for example, the N-terminal α-amino group (Nα group) of the amino acid serving as the terminal of the target peptide is changed to 9-fluorenylmethyloxycarbonyl group (Fmoc group) or t- Protect with a protecting group such as a butyloxycarbonyl group (Boc group) and activate the carboxyl group of the amino acid by esterification with 1-hydroxybenzotriazole (HOBt) or the like directly or depending on the case , Chloromethyl resin (chloromethylated styrene-divinylbenzene copolymer) having a chloromethyl group or oxymethyl group through a spacer, Super Acid Labeille Polyethylene Glycol (Fmoc-NH-SAL (super acid-labile) resin , Super Acid Labile Polyethylene Glycol (Fmoc-NH-SAL-PEG (polyethyleneglycol) resin, etc. Then, the Nα-protecting group is removed from the fixed protected amino acid using a base such as piperidine, and the amino group of the fixed protected amino acid and the adjacent second protected amino acid (Nα-amino group) are fixed. And the side chain functional group in the same manner with a protecting group, and the carboxyl group of the amino acid) is condensed to form a peptide bond, and the Nα-protecting group of the obtained second protected amino acid is eliminated, and other amino acids are similarly removed. The desired peptide chain can be prepared by sequential condensation, and then the insoluble resin and amino acid Nα-amino group and side chain functional group protecting groups can be eliminated and synthesized.

  In addition to the above active ester method, the peptide of the present invention can be chemically synthesized by methods such as the azide method, acid chloride method, acid anhydride method, mixed acid anhydride method, DCC method, and oxidation-reduction method. .

  The peptide of the present invention may be a derivative in which a part of the amino acid sequence is chemically modified, or a derivative substituted with other amino acids including non-natural amino acids. As used herein, “chemical modification” refers to the reaction of a chemical reagent with a peptide, mainly changing the chemical structure of the side chain of an amino acid residue (Motonori Ohno, Yuichi Kanaoka, Fumio Sakiyama, Maeda). Hiroshi, Biochemical Experimental Method 12, Chemical Modification of Protein (above), Academic Publishing Center). In this invention, as a derivative in which a part of the amino acid sequence is chemically modified, for example, the amino acid α-carbon is methylated, or the amino acid C-terminal is amidated with an alkylamine such as ethylamine, ethanolamine, or ethylenediamine. And those modified on the amino acid side chain, for example, those obtained by acylating the ε-amino group of lysine. Furthermore, since the peptide derivative of this invention is particularly useful for pharmaceutical use, it is preferably a pharmacologically acceptable derivative. The modified portion of the peptide containing a chemically modified amino acid sequence does not affect the original activity of the peptide but acts as another effect (Yamaguchi, H. et al., Biosci. Biotechnol. Biochem., 67 ( 10), 2269-2272, 2003).

  The peptides of the present invention also include salt forms. The salt is preferably a physiologically acceptable acid addition salt or basic salt. Acid addition salts include, for example, salts with inorganic acids such as hydrochloric acid, phosphoric acid, hydrobromic acid, sulfuric acid, or acetic acid, formic acid, propionic acid, fumaric acid, maleic acid, succinic acid, tartaric acid, citric acid, apple And salts with organic acids such as acid, oxalic acid, benzoic acid, methanesulfonic acid, and benzenesulfonic acid. Examples of basic salts include salts with inorganic bases such as sodium hydroxide, potassium hydroxide, ammonium hydroxide and magnesium hydroxide, and salts with organic bases such as caffeine, piperidine, trimethylamine and pyridine. . Of these salts, pharmaceutically acceptable salts are preferred. These salts can be prepared using a suitable acid such as hydrochloric acid or a suitable base such as sodium hydroxide. For example, it can be prepared by treatment using standard protocols in water or in a liquid containing an inert water-miscible organic solvent such as methanol, ethanol or dioxane.

  The peptide, derivative thereof and salt thereof according to the present invention have antibacterial activity, antiviral activity, antiparasitic activity, or a combination thereof, against bacteria, fungi, viruses, parasites and the like.

Examples of bacteria to which the peptide of the present invention is applied include various gram-positive bacteria and gram-negative bacteria. Examples of such bacteria include the following.
Streptococcus sp., For example S. pneumoniae, Group A B hemolytic streptococci (S. pyogenes), Group B Streptococcus (S. agalactiae), S. milleri group, green streptococci (viridans) streptococci), oral streptococci, etc .; Staphylococcus sp., for example, S. aureus, S. epidermidis, coagulase negative streptococcus; , S. haemolyticus, S. saprophyticus, S. capitis, S. caprae, S. lugdunensis, S. saccharolyticus, S. warneri, S. homini, S. cohnii; S. xylosus, S. simulans, S. schleiferi), etc. Corynebacterium sp., Such as C. diphtheriae, etc .; Enterococcus sp., Such as E. faecalis, E. faecium, etc .; Acinetobacter sp., Such as Acinetobacter sp. Bow A. baumanii, A. calcoaeticus, etc .; Citrobacter sp .; Enterobacter sp .; Escherichia sp., For example, E. coli, etc .; Klebsiella (Klebsiella sp.);
Morganelia sp .; Proteus sp .; Providencia sp .; Salmonella sp., Such as S. typhi, S. typhimurium; Serratia sp. Shigella sp .; Pseudomonas sp., For example P. aeruginosa, P. acnes; Yersinia sp., For example Y. pestis (Y. pestis), pseudotuberculosis Y. pseudotuberculosis, Y. enterocolitica; Franciscella sp .; Pasturella sp .; Vibrio sp., such as Vibrio cholerae ( V. cholerae), V. parahemolyticus; Campylobacter sp., Such as C. jejuni; Haemophilus philus sp.), for example H. influenzae, H. ducreyi; Bordetella sp., for example B. pertussis, B. bronchiseptica, para B. parapertussis; Brucella sp .; Neisseria sp., Such as N. gonorrhoeae, N. meningitidis, etc .; Legionella sp. Legionella pneumophila; Listeria sp., Eg, L. monocytogenes; Mycoplasma sp., Eg, Mycoplasma hominis, pneumonia mycoplasma (M. pneumoniae); Mycobacterium sp., for example M. tuberculosis, M. leprae; Treponema sp., for example syphilis trepo T. parallidum; Borrella sp., For example, B. burgdorferi; Leptospirae sp .; Rickettsia sp., For example, spotted fever rickettsia (R. rickettsii), rash fever rickettsia (R. typhi); Chlamydia sp., for example, C. trachomatis, pneumonia chlamydia (C. pneumoniae), parrot disease Chlamydia (C. psittaci); Helicobacter sp .), Eg H. pylori, etc .; Plasmodia sp., Eg P. felciparum, Trypanosoma sp., Eg, T. brucer; cystosomes (Schistosomes); Endoparasitic amoeba (Entaemoeba sp.), Cryptococcus sp., Candida sp., Eg Candida Albicans (C. albicans), Candida Jeikeumu (Candida jeikeium), Candida Aferumentasu (C. afermentas) and the like.

  In addition, the peptide according to the present invention is a coagulase-negative staphylococci (CNS) such as Staphylococcus epidermidis which is a gram-positive cocci resident in human healthy skin among the above bacteria. And the above-mentioned activity against Corynebacterium spp. (Corynebacterium spp.), Which is a gram-positive bacilli, and Acinetobacter spp. These bacteria are harmless in plains, but are caused by bloodstream infections related to artificially inserted valves such as heart replacement valves, central nervous system shunts, and vascular catheters, especially in susceptible patients Sometimes. CNS is a major causative agent in vascular catheter-related infections and sometimes develops to endocarditis and meningitis. Many pathogenic strains produce alpha hemolysin, DNA-degrading enzymes, and staphylokinase that activates plasminogen in the serum to produce plasmin. Biofilm formation is also related to pathogenicity, and Staphylococcus epidermidis may adhere to plastic surfaces such as catheters and form biofilms. The peptides of this invention are also effective against such biofilms. It is.

  Further, the peptide according to the present invention is, for example, a multidrug resistant gram positive bacterium (for example, Methicillin-Resistant Staphylococcus aureus (MRSA), methicillin sensitive yellow grape Staphylococcus aureus (MSSA), (Vancomycin-Intermediate Staphylococcus aureus: VISA), Vancomycin-Resistance Staphylococcus aureus (VRSA), Vancomycin-Resistant Enterobacter, etc. It is also effective against bacteria (for example, multi-drug resistant Helicobacter, Shigella, Salmonella, etc.).

  The peptides according to the invention also have antifungal activity against Candida, Aspergillus, Cryptococcus, Mucor and many other fungi.

  The peptides according to the present invention are viruses such as alphavirus, arenavirus, bunyavirus, coronavirus, enterovirus, filovirus, flavivirus, hantavirus, HLTV-BLV, influenza virus, norovirus, lentivirus, lassavirus, para RNA viruses such as myxovirus, reovirus, rhinovirus, rotavirus, SARS virus, hepatitis B virus, hepatitis C virus; adenovirus, cytomegalovirus, hepadnavirus, molsypox virus, orthopox virus, papilloma It has antiviral activity against DNA viruses such as viruses, parvoviruses, polyoma viruses, simplex viruses, varicella-zoster virus.

  Furthermore, the peptide of the present invention has activity against protozoa and parasites. Examples of the protozoa include amoeba, malaria, toxoplasma, cryptosporidium, and babesia. Parasites include, for example, roundworms, helminths, helminths, fecal nematodes, filariae, anisakis, jaw-and-mouthworms, nematodes, schistosomiasis, liver flukes, Yokogawa flukes, lung flukes, etc. , And tapeworms such as striped worms.

Therefore, the peptide according to the present invention can be effectively applied to infectious diseases caused by bacteria, fungi, viruses, parasites and the like.
Representative examples of bacterial infections include actinomycosis, anthrax, treponema, campylobacterosis, cholera, gas gangrene, enterobacterial infection, hemophilus infection, mycobacterium infection, leptospirosis, listeriosis , Lyme disease, plague, pneumococcal infection, Pseudomonas infection, Salmonella infection, bacterial dysentery, staphylococcal infection, streptococcal infection, tetanus, toxic shock syndrome, savage disease, typhoid, sexually transmitted disease (syphilis) Gonococcal infection, non-gonococcal infection, chlamydia cervicitis, trichomoniasis, etc.).

  Typical examples of infections caused by fungi include aspergillosis, blastosomiasis, canditasis, coccidioidomycosis, cryptococcosis, histoblastoma, mucormycosis, baracocidioidomycosis, sporotorosis and the like.

  Representative examples of viral infections include colds, influenza, herpes simplex virus infection, shingles, Epstein-Barr virus infection, cytomegalovirus infection, hemorrhagic fever, hantavirus infection, yellow fever, Examples include dengue fever, severe acute respiratory syndrome, and human immunodeficiency virus (HIV) infection.

  Representative infectious diseases caused by parasites include, for example, amebiasis, roundworm, babesiosis, cryptosporidiosis, giardiasis, helminthiasis, malaria, helminthiasis, schistosomiasis, tapeworm infection, toxocariasis, toxoplasma Disease, trichinosis, trichuriasis and the like.

  Furthermore, the peptide according to the present invention can also be used as an antibacterial composition in combination with an antibiotic, an antifungal agent, an antiviral agent and / or an antiparasitic agent, and an antibacterial drug. By combining the peptides of the present invention with antibiotics, antifungal agents, antiviral agents and / or antiparasitic agents, it is possible to obtain a great advantage that the mutual efficacy can be complemented or enhanced.

  Any antibiotic can be used in the present invention, and is not particularly limited. For example, penicillin antibiotics, cephalosporin antibiotics, cephamycin antibiotics, oxacephem Β-lactam antibiotics, aminoglycoside antibiotics, oxazolidinone antibiotics, glycopeptide antibiotics, polypeptides selected from the group consisting of antibiotics, carbacephem antibiotics, carbapenem antibiotics and monobactam antibiotics Antibiotics, chloramphenicol antibiotics, tetracycline antibiotics, macrolide antibiotics, lincomycin antibiotics, quinolone antibiotics, ketolide antibiotics, glycylcycline antibiotics and other antibiotics Or sulphur It is preferable to appropriately select at least one of from the group consisting of. Of course, antibiotics other than those described above can be effectively applied.

  Such antibiotics include, for example, penicillin G, penicillin V, ampicillin, amoxicillin, bacampicillin, cloxacillin, dicloxacillin, methicillin, nafcillin, oxacillin, azulocillin, carbenicillin, mezlocillin, piperacillin, ticarcine, etc .; Cefachlor, cefamandole, cefotetan, cefoxinone, cefmethazole, cefoniside, cefprodil, cefuroxime, etc .; Cefoxitin, Fotetan, cefbuperazone, cefminox, cefmetazole, etc .; latamoxef, flokimosef, etc .; imipenem / cilastanem, ertabenem, meropenem, faropenem, panipenem, etc .; , Spectinomycin, tobramycin, netilmycin, neomycin (fradiomycin) etc .; vancomycin, teicoplanin etc .; linezolid etc .; chloramphenicol, thianphenicol etc .; oxytetracycline, tetracycline, demeclocycline, doxycycline, minocycline etc .; Clarice Romaishi , Troleandomycin, roxithromycin, azithromycin, dirithromycin, acetylspiramycin, kitasamycin, josamycin, midecamycin, roquitamycin, midecamycin acetate, etc .; quinolone, nalidixic acid, enoxacin, ofloxacin, gatifloxacin, garenoxacin, ciprofloxacin , Sparfloxacin, tosufloxacin, trovafloxacin, norfloxacin, fleroxacin, moxifloxacin, levofloxacin, lomefloxacin, etc .; lincomycin, clindamycin, etc .; bacitracin, colistin, polymyxin B, etc .; tethromycin, etc .; sulfa Chlorpyridazine, sulfasalazine, sulfadiazine, sulfadimethoxine, sulfacetamide, sulf Doxin, sulfamethizole, famethoxazole, sulfisoxazole, trimethoprim-sul, mafenide, etc .; tigesancrine, etc .; isoniazid, etc .; virginiamycin, quinupristin / dalfopristin, ethambutol, phosphomybristin / dalfobristin, rifampicin, subex Tinomycin is mentioned.

  Examples of antifungal agents that can be used in the present invention include polyene macrolides such as amphotericin and nystatin, pyrimidines such as flucytosine, miconsol, ketoconazole, clotrimazole, econazole, isoconazole, sulconazole, oxyconazole, croconazole, Ibonoids such as bibonazole, neticonazole and lanconazole, triazoles such as fluconazole, itraconazole and voriconazole, candins such as Micafungin, allylamines such as terbinafine hydrochloride, thiocarbamates such as liranaphthalate, benzylamines such as butenafine, amorolfine, etc. And other antibiotics such as morphomin and griseofulvin.

  Antiviral agents that can be used in the present invention include, for example, acyclovir, valacyclovir, penciclovir, famciclovir, valganciclovir hydrochloride, cidofovir, trifluridine, fomivirsen, vidarabine, zalcitabine, idoxuridine, sorivudine, brivudine, stavudine, zidovudine Edoxudine, sodium ganciclovir, sodium foscarnet, oseltamivir, zanamivir, amantadine, rimantadine, didanosine, ribavirin, interferon-α and the like.

  Antiparasitic drugs that can be used in the present invention include, for example, metronidazole, mebendazole, albendazole, birantel vamorate, quinine, clindamycin, furazolidone, chloroquine, mefloquine, doxycycline, atobacon, proguanil, praziquantel, diethylcarbamazine, sulfadiazine , Pyrimethamine, trimethoprim-sulfamethoxazole (ST combination), ivermectin, niclosamide, and bithionol.

  The peptide, antibacterial composition and antibacterial drug according to the present invention can be administered in various ways such as systemic or topical administration, such as parenteral or oral, such as subcutaneous, intradermal, intravenous and intraperitoneal, depending on the purpose of the treatment. Each can be administered by route. In addition, the dosage form for administering the peptide, antibacterial composition and antibacterial medicine of the present invention can be applied by appropriately selecting various dosage forms which are usually used in the pharmaceutical field and the like. Examples of the mold include solutions, suspensions, emulsions, injections such as solid preparations for use, tablets, capsules, granules, powders, pills, solutions, syrups, ointments, gels External powders, sprays, creams, inhaled powders, eye drops, eye ointments, suppositories and the like can be used. These preparations can be prepared according to a conventional method.

  The preparation of the peptide, antibacterial composition and antibacterial drug of the present invention as described above is a preparation used in the application field in addition to the active ingredient of the peptide, antibacterial composition and antibacterial drug. A carrier for use may be contained. The preparation carrier is not particularly limited and can be appropriately selected depending on the purpose of use. For example, excipient, extender, binder, lubricant, lubricant, colorant, wetting Agents, dispersants, disintegrants, buffers, isotonic agents, preservatives, surfactants, preservatives, flavoring agents, soothing agents, stabilizers, solubilizers, and the like can be used.

  In addition, the peptide, antibacterial composition and antibacterial pharmaceutical preparation of the present invention may contain other pharmaceutically acceptable carriers and / or additives in addition to the above carriers. Carriers / additives include, for example, water, pharmaceutically acceptable organic solvents, collagen, sodium alginate, water-soluble dextran, sodium carboxymethyl starch, pectin, xanthan gum, gum arabic, casein, gelatin, agar, glycerin, propylene Examples include glycol, polyethylene glycol, petrolatum, paraffin, stearyl alcohol, stearic acid, human serum albumin, mannitol, sorbitol, and lactose. The carrier / additive to be used can be appropriately selected from the above or in combination with the above according to the dosage form of the present invention.

  When the peptide of the present invention is administered as a medicament such as an antibacterial agent, the dosage can be appropriately changed over a wide range depending on the target disease, the type and age of the administration target, the administration route, the number of administrations, and the like. The effective amount of the active ingredient is generally in the range of about 1-1000 mg, preferably about 5-500 mg, more preferably about 10-200 mg per day for an adult (assuming a body weight of 60 kg). It is good to administer it once a day or several times a day or more.

  The peptide of the present invention is also effective against nosocomial infections and opportunistic infections that are problematic in hospital facilities, etc., and in the prevention and treatment of such infections, particularly in combination with antibiotics as necessary. It is preferable to apply. Furthermore, the peptide of the present invention is used in combination with antibiotics as necessary, medical devices and prostheses causing nosocomial infections, such as catheters, stents, tubes, probes, cannulas, artificial heart valves, dialysis membranes When applied to filters, films, non-woven fabrics, surgical sutures, needles, surgical instruments, etc., such medical devices and prostheses can be sprayed or coated in the form of solutions, creams, ointments, etc. It is good to use it, such as tinging, and in that case, it is good to apply in the range of 0.1-5%, preferably 0.5-2%.

  The dose and the number of doses of the drug that can be used in combination with the peptide according to the present invention can be administered in the same manner as when such drug is administered alone, but the amount is determined in consideration of the subject to be administered in combination. It is better to increase or decrease appropriately. For example, since the general doses of the following drugs are as follows, when the following drugs are used in combination, it is preferable to select and administer the dose appropriately.

Ambicillin: 250-500mg / 8hr
Amoxicillin: 250-500mg / 8hr
Methicillin: 100-300 mg / day Cefazolin: 100-400 mg / day Ceftriaxone: 4 g / day Cefotaxime: 12 g / day Ciprofloxacin: 400-1500 mg / day gentamicin: 3 mg / kg / day Tobramycin: 3 mg / kg / day Imipenem: 1500mg / kg
Piperacillin: 24 g / day vancomycin, teicoplanin: 6-30 mg / kg / day Streptomycin: 500 mg-1 g / 12 hr
Metronidazole: 2-4 g / day Tetracycline: 500 mg / 5 hr
Rifampin: 600 mg / day fluconazole: 150-400 mg / day Acyclovir: 200-400 mg / day Ribapilin: 20 mg / day amantadine, rimantadine: 200 mg / day chloroquine: 800 mg / day

Peptides having the amino acid sequences represented by the chemical formulas [III] to [VIII] were synthesized according to a solid phase synthesis method (Fmoc method) using an automatic peptide synthesizer (PTI-PS3: Aloka Co., Ltd.). In this solid phase synthesis method, Fmoc-NH-SAL-PEG resin (manufactured by Watanabe Chemical Co., Ltd.) is used as the solid phase carrier, and 9-fluorenylmethyloxycarbonyl (Fmoc) -amino acid (Fmoc-AA) is used as the protected amino acid. -OH), O-benzotriazol-1-yl-N, N, N ', N'-tetramethyluronium hexafluorophosphate (HBTU) and 1-hydroxybenzotriazole (HOBT) as condensing agents, N as solvent , N-diisopropylethylamine (DIEA) was used.
First, 0.4167 g of Fmoc-NH-SAL-PEG resin (0.1 mmol) was swollen with N-methyl-2-pyrrolidone (NMP) for about 30 minutes. This resin was put into a container, and 0.1517 g of HBTU and 0.0613 g of HOBT-OH were added thereto. Furthermore, the amount of amino acid derivative (Fmoc-AA-OH) of the target peptide shown in Table 1 below was added to this container. The reaction molar ratio of Fmoc-AA-OH: HBTU: HOBT: DIEA: H2N-resin was adjusted to 3: 3: 3: 6: 1.
The containers containing the amino acid derivatives thus prepared were arranged in the order of RVG-9R sequence, loaded on the rotary table of the automatic peptide synthesizer, and finally an empty container was loaded.

As solvents, NMP as PRI-SOLV1, NMP as PRI-SOLV2, ACT: 0.9M DIPEA (DIPEA 78 ml + NMP 422 ml), DEP: 20% piperidine (piperidine 100 ml + NMP 400 ml) were prepared and used.
First, the remaining liquid remaining in the tube was discarded by a vent, and the solvent was replenished to normal pressure. Next, the container was pressurized, the chirp was filled with the solution with the prime, and the synthesis was started according to the program of the automatic peptide synthesizer.

  In order to deprotect the protecting group of the peptide obtained as described above, the peptide was transferred to an eggplant flask and trifluoroacetic acid (TFA) solution (TFA 9 ml: thioanisole 500 μl: m-cresol 500 μl: ethanediol 250 μl) ) Was added, and a stir bar was added and stirred for 1 hour. This solution was folded and filtered in a tall beaker and washed with a small amount of TFA. An appropriate amount of ice-cold diethyl ether was added to the filtrate and the mixture was ice-cooled. The white precipitate generated in this manner was filtered under reduced pressure, the residue on the filter paper was transferred to a tall beaker, water was added, dissolved by irradiating with ultrasonic waves, and pleated and filtered in an eggplant flask. The resulting filtrate was lyophilized with liquid nitrogen. This lyophilized product was powdered and stored at -20 ° C.

  The peptide obtained above was purified using high performance liquid chromatography (HPLC). The powdered peptide was dissolved in MillQ water, passed through a 0.44 μl filter to prevent large impurities from entering the column, and then injected into the HPLC with a syringe. The sample was collected at the peak of the target peptide detected by the detector, and the collected solution was dried by a speed feed back centrifuge and stored at -20 ° C. As an eluent as a mobile phase, 0.1% TFA aqueous solution was used as A liquid, acetonitrile + 0.1% TFA liquid as B liquid, and 100% methanol as C liquid. The concentration gradient of the eluent in HPLC was set according to the program shown in Table 2 below.

The molecular weight of each peptide eluted as described above was determined based on MALDI-TOF / MS mass spectrometry using a KRATOS mass spectrometer (KOMPACT MALDI III: Shimadzu Corporation). That is, a powdery peptide sample was dissolved in a 0.1% TFA aqueous solution, and 5 μl of α-cyano-4-hydroxycinnamic acid (CHCA: 10 mg / ml) was added to and mixed with the sample solution. This mixed solution was dropped on a spot on the sample plate. As a result, it was confirmed that the target peptide was synthesized and purified (Table 3). In addition, the conditions of mass spectrometry are as follows.
Matrix: α-cyano-4-hydroxycinnamic acid (CHCA)
Measurement mode: Linear reflector mode

About each of the peptide synthesize | combined in Example 1, the antibacterial activity was measured in accordance with the method as described in a trace amount liquid dilution method (based on the method of the Japan Chemotherapy Society).
After culturing on the plate at 37 ° C. for 5 days, the number of viable cells was measured. Antibacterial activity is the minimum inhibitory concentration (MIC ₅₀ : μg / ml), effective concentration that inhibits 50% cytotoxicity due to HIV infection (EC ₅₀ ; 50% effective concentration), and cytotoxicity is 50% cytotoxic concentration by the test substance ( CC ₅₀ ; 50% cytotoxic concentration). Also, effective coefficient (Selectivity Index; SI) was calculated as CC ₅₀ / EC _50.
The test strains were Staphylococcus. And P. acnes were used.

The amino acid sequence of the test peptide is as follows.
Peptide [III]: IleLeuArgWsProWsWsProWsArgArgLys (SEQ ID NO: 3)
Peptide [IV]: IleLeuArgTrpProTrpTrpProTrpArgArgLysAlaAlaAlaAla (SEQ ID NO: 4)
Peptide [V]: IleLeuArgTrpAlaTrpLysAlaTrpArgArgTrpAlaAlaAlaAla (SEQ ID NO: 5)
Peptide [VI]: Ac-IleLeuArgWsProWsWsProWsArgArgLysAlaAlaAlaAla (SEQ ID NO: 6)
Peptide [VII]: IleLeuArgTrpAlaTrpLysAlaTrpArgArgTrpAlaArgAlaAla (SEQ ID NO: 7)
Peptide [VII]: Ile-Leu-Arg-Trp-Pro-Trp-Trp-Pro-Trp-Arg-Arg-Lys-NH ₂ (SEQ ID NO: 8)

The results of MIC50 are shown in the table below.

The results of MBC50 are shown in the table below.

With respect to the minimum inhibitory concentration (MIC50) and the minimum bactericidal concentration (MBC50), respective reduction rates (%) when omiganane is set to 100 are shown in Tables 6 and 7, respectively.

The composition example which formulated the peptide of this invention into the antibacterial agent (tablet) is shown.
Peptide [III]: 100 mg
Lactose: 670mg
Potato starch: 150mg
Crystalline cellulose: 60mg
Light anhydrous silicic acid: 50mg
After mixing the above ingredients, adding a solution of hydroxypropylcellulose 30 mg in methanol (hydroxypropylcellulose 10% by weight), kneading and granulating, extruding into a 0.8mm diameter screen, drying, and drying Tablets were obtained by adding 15 mg of magnesium stearate and compressing 200 mg each.

The composition example which formulated the peptide of this invention in the bacterial infection treatment agent (capsule agent) is shown.
Peptide [IV]: 100 mg
Lactose: 80mg
The above ingredients were mixed uniformly and filled into hard capsules to obtain capsules.

The composition example which formulated the peptide of this invention into the bacterial infection treatment agent (injection) is shown.
Peptide [V]: 30mg
The above components were dissolved in 2 ml of 5% mannitol aqueous solution, filtered aseptically, and sealed in an ampoule.

The composition example which formulated the peptide of this invention into the bacterial infection treatment agent (injection for dissolution at the time of use) is shown.
(A) Peptide [V] (lyophilized product): 30 mg (encapsulated in an ampoule)
(B) 2 ml of sterile filtered PBS (enclosed in an ampoule)
The above-mentioned (A) and (B) were used as one set to produce an injection for dissolution at the time of use. At the time of use, (A) can be dissolved in (B) and used.

The composition example which formulated the peptide of this invention into the antibacterial agent (ointment) is shown.
Peptide [III]: 10mg
Sorbitan monostearate: 7mg
Polyoxyethylene sorbitan monostearate: 7mg
Isopropyl palmitate: 37 mg
Vaseline: 37mg
Liquid paraffin: 37mg
Cetanol: 50mg
Glycerin: 70mg
Magnesium stearate: 2mg
Purified water was added to the above ingredients to make 1 g of cream.

  In substantially the same manner as in Example 9, 1 g of cream was prepared using each of peptides [IV] to [VIII] instead of peptide [III].

The composition example which formulated the peptide of this invention into the antibacterial agent (ointment) is shown.
Peptide [VIII]: 10mg
Penicillin (ampicillin): 10mg
Sorbitan monostearate :: 7mg
Polyoxyethylene sorbitan monostearate: 7mg
Isopropyl palmitate: 37 mg
Vaseline: 37mg
Liquid paraffin: 37mg
Cetanol :: 50mg
Glycerin: 70mg
Magnesium stearate: 2mg
Purified water was added to the above ingredients to make 1 g of cream.

Peptide [III]: 0.5%
Vaseline: 26.3%
Paraffin: 5.3%
Cetostearyl alcohol: 2.1%
Propylene glycol: 10.5%
Polyoxyethylene / polyoxypropylene glycol ether: 3.2%
Purified water: 52.1%
The said component was mixed and the skin external preparation was prepared.

  In substantially the same manner as in Example 11, skin external preparations were prepared using peptides [IV] to [VIII] in place of peptide [III].

  The peptide according to the present invention can be used particularly for the prevention of infection in various medical and environmental health fields, and the treatment and reduction of the incidence of infectious diseases.

Sequence number 1: Synthetic peptide Sequence number 1: Xaa represents Trp or thiotryptophan (location: 4).
Sequence number 1: Xaa represents Pro or Ala (location: 5).
Sequence number 1: Xaa represents Trp or thiotryptophan (location: 6).
Sequence number 1: Xaa represents Trp, thiotryptophan, or Lys (location: 7).
Sequence number 1: Xaa represents Pro or Ala (location: 8).
Sequence number 1: Xaa represents Trp or thiotryptophan (location: 9).
Sequence number 1: Xaa represents Lys or Trp (location: 12).
Sequence number 1: Xaa represents Ala- (Ala / Arg) -Ala-Ala (location: 13).
SEQ ID NO: 2: Synthetic peptide SEQ ID NO: 2: Xaa represents Ala or Arg (location: 2).
SEQ ID NO: 3: Synthetic peptide SEQ ID NO: 3: Xaa represents thiotryptophan (location: 4).
Sequence number 3: Xaa represents thiotryptophan (location: 6).
Sequence number 3: Xaa represents thiotryptophan (location: 7).
Sequence number 3: Xaa represents thiotryptophan (location: 9).
SEQ ID NO: 4: Synthetic peptide SEQ ID NO: 5: Synthetic peptide SEQ ID NO: 6: Synthetic peptide SEQ ID NO: 6: Xaa represents thiotryptophan (location: 4).
Sequence number 6: Xaa represents thiotryptophan (location: 6).
Sequence number 6: Xaa represents thiotryptophan (location: 7).
Sequence number 6: Xaa represents thiotryptophan (location: 9).
SEQ ID NO: 7: synthetic peptide SEQ ID NO: 8: synthetic peptide

Claims (10)

Formula [I]:
Ile-Leu-Arg-X1-X2-X3-X4-X5-X6-Arg-Arg-X7- (X8) n [I]
(Where X1 is tryptophan (Trp) or chemical formula [II]:

Thiotryptophan (Ws) represented by
X2 means proline (Pro) or alanine (Ala);
X3 means tryptophan (Trp) or thiotryptophan (Ws);
X4 means tryptophan (Trp), thiotryptophan (Ws) or lysine (Lys);
X5 means proline (Pro) or alanine (Ala);
X6 means tryptophan (Trp) or thiotryptophan (Ws);
X7 means lysine (Lys) or tryptophan (Trp);
X8 means Ala-X9-Ala-Ala (where X9 means Ala or Arg);
n means 0 or 1; and each amino acid means L form or D form;
Where X1, X3, X4 and X6 are each tryptophan (Trp); X2 and X5 are each proline (Pro); X7 is lysine (Lys) and each amino acid is L-form In this case, n means 1)
Or a derivative thereof or a salt thereof. The peptide according to claim 1 or a derivative thereof, or a salt thereof, wherein the peptide has any amino sequence represented by the following formulas [III] to [VIII].

Ile-Leu-Arg-Ws-Pro-Ws-Ws-Pro-Ws-Arg-Arg-Lys [III]
Ile-Leu-Arg-Trp-Pro-Trp-Trp-Pro-Trp-Arg-Arg-Lys-Ala-Ala-Ala-Ala [IV]
Ile-Leu-Arg-Trp-Ala-Trp-Lys-Ala-Trp-Arg-Arg-Trp-Ala-Ala-Ala-Ala [V]
Ile-Leu-Arg-Ws-Pro-Ws-Ws-Pro-Ws-Arg-Arg-Lys-Ala-Ala-Ala-Ala [VI]
Ile-Leu-Arg-Trp-Ala-Trp-Lys-Ala-Trp-Arg-Arg-Trp-Ala-Arg-Ala-Ala [VII]
Ile-Leu-Arg-Trp-Pro-Trp-Trp-Pro-Trp-Arg-Arg-Lys (all in D-form) [VIII]
(Wherein Ws has the same meaning as above).   A peptide consisting of 12 to 40 amino acids containing the amino sequence of the peptide according to claim 1 or 2, a derivative thereof, or a salt thereof.   A peptide comprising 12 to 30 amino acid sequences containing the amino sequence of the peptide according to claim 1 or 2, or a derivative thereof, or a salt thereof.   A peptide consisting of 12 to 20 amino acid sequences containing the amino sequence of the peptide according to claim 1 or 2, or a derivative thereof, or a salt thereof.   A peptide consisting of 12 to 16 amino acids containing the amino sequence of the peptide according to claim 1 or 2, or a derivative thereof, or a salt thereof.   7. The peptide according to claim 1, wherein the peptide has at least one activity selected from the group consisting of antibacterial activity, antiviral activity and growth inhibitory activity against bacteria, fungi and / or viruses. Or a derivative thereof, or a salt thereof.   The peptide or derivative thereof or a salt thereof according to any one of claims 1 to 7, wherein the peptide has a growth inhibitory activity against Gram-positive bacteria.   The peptide according to any one of claims 1 to 8, or a derivative thereof, or a salt thereof, and at least one selected from the group consisting of antibiotics, antifungal agents, antiviral agents, and antiparasitic agents. The composition characterized by the above-mentioned.   A pharmaceutical comprising the peptide according to any one of claims 1 to 8, a derivative thereof, or a salt thereof, and / or the composition according to claim 9.