JPWO2013039190A1 - Peptide-containing polymer and method for immobilizing peptide on fiber - Google Patents

Abstract

An object of the present invention is to provide a polypeptide-containing polymer that can be used in the production of antimicrobial fibers. The object of the present invention is also to a fiber of a polypeptide that can immobilize an antimicrobial polypeptide to a fiber by a relatively easy means and has little damage to the fiber, but can maintain the fixation of the polypeptide by repeated use. It is to provide an immobilization method. INDUSTRIAL APPLICABILITY According to the present invention, an antibacterial polypeptide (beetle defensin-modified polypeptide) -containing polymer that can be used for the production of antibacterial fibers is provided. In addition, the antimicrobial polypeptide can be easily immobilized on the fiber by the method of the present invention.

Description

  The present invention relates to a novel peptide-containing polymer. The present invention also relates to a novel method for immobilizing peptides on fibers.

  In recent years, drug-resistant bacteria that are resistant to antibiotics have spread in developed countries, causing mass infections and nosocomial infections. In particular, more than 20,000 cases of infection with methicillin-resistant Staphylococcus aureus (MRSA) resistant to methicillin are reported annually in Japan. Infection routes are complex and difficult to identify, but bed sheets and lab coats used in hospitals are considered infection risk factors.

  In order to solve these problems, the inventors have developed an antibacterial fiber in which a modified peptide derived from beetle defensin (Non-patent Document 1) invented by Ishibashi et al. A molecular chain called a spacer was bound to cotton, and amino acid synthesis was carried out starting from the terminal amino group, and a 9-residue modified peptide was synthesized on cotton. As a result of the antibacterial test, MRSA showed strong antibacterial activity, and it was revealed that the activity was maintained even after repeated use (Patent Document 1, Non-Patent Document 2). However, this method involves complicated processing methods such as fiber pretreatment, and is difficult to implement with existing fiber processing techniques. In addition, since the condensation reaction of amino acids and the deprotection reaction of protecting groups are all carried out on cotton, the mechanical strength of the fiber itself is also lowered, so that it was necessary to develop a new processing method for practical use.

  In addition, an antibacterial multiple antigen peptide in which an antibacterial peptide is linked to the N terminal of a branched peptide in which 15 Lys are linked, which is a positively charged antibacterial peptide in an aqueous solution, is fixed to the fiber with a resin-based fixing agent. Has been reported (Patent Document 2). However, there is a possibility that a part of the peptide fixed with the resin-based fixing agent may lose its activity when it is fixed, and it is necessary to fix more antibacterial peptides than necessary. In addition, peptides that can be fixed with a resin-based fixing agent are limited.

  Therefore, there is a demand for a new and simple method for immobilizing peptides on fibers, in which the activity of the peptides immobilized on the fibers is sufficiently maintained in use and does not involve an amino acid synthesis reaction on the fibers. It was.

  Also reported is a physiologically active substance-immobilized substrate characterized in that it has a polymer compound on the surface comprising a unit having a phosphorylcholine group, a unit having a hydrophobic group, and a unit having an aldehyde group or a maleimide group. (Patent Document 3). However, it is not immobilization of the peptide to the fiber.

JP 2010-184022 A JP 2000-45182 A JP 2010-117189 A Japanese Patent No. 3273314 JP 2010-184022 A

Ishibashi et al., Eur. J. Biochem, 1999 Dec; 266 (2): 616-23 Nakamura et al., Biomacromolecules, 2011, 12 (5): 1540-1545

  An object of the present invention is to provide a polymer containing an antibacterial peptide (a modified peptide derived from beetle defensin having antibacterial properties) that can be used for the production of antibacterial fibers. The object of the present invention is also a new immobilization capable of immobilizing peptides, particularly antibacterial peptides, to fibers by relatively easy means, with less damage to the fibers, and capable of maintaining the immobilization of peptides even after repeated use. Is to provide a method. The object of the present invention is also to provide an antimicrobial fiber having an antimicrobial peptide.

As a result of diligent efforts, the present inventors can fix the peptide to the fiber by a relatively easy method, maintain the peptide activity on the fiber, and maintain the peptide activity even after repeated washing of the fiber. I found what I could do and completed the present invention.
If the peptide-containing polymer of the present invention is used, the peptide can be immobilized on the fiber by a simple method. More specifically, by using the polymer of the present invention, the peptide can be immobilized on the fiber without complicated pretreatment of the fiber and without peptide synthesis on the fiber.
The present invention also relates to a method for immobilizing peptides on fibers. By using the method of the present invention, the peptide can be immobilized on the fiber by a simple method.

In one embodiment of the present invention, a polymer containing an antibacterial peptide (modified peptide derived from beetle defensin having antibacterial properties) is provided, and the polymer of the present invention can be immobilized on a fiber by a simple method.
In one embodiment of the present invention, a modified peptide derived from beetle defensin having antibacterial properties is conjugated with a linker in the polymer, such as a maleimide group, to prepare a peptide-containing polymer, and the antibacterial peptide is coated by coating it on a fiber. Can be immobilized on fiber.

In another embodiment of the present invention, after the fiber is coated with a linker having a binding property to a peptide, such as a polymer having a maleimide group, the fiber is added to a solution containing a peptide, particularly a modified peptide derived from beetle defensin having antibacterial properties. So that the peptide can be immobilized on the fiber.
In another aspect of the present invention, an antibacterial fiber in which a modified peptide derived from beetle defensin having antibacterial properties provided as described above is immobilized can be provided.

That is, the following invention is provided.
(1) The following formula (1)

（式中Ｒ₁は、水素原子又はメチル基であり、Ｕ₁は、単結合、Ｏ又はＮＨであり、Ｖは、単結合又はスペーサーであり、Ｗは、単結合或いはアミド基、チオエーテル基、チオエステル基、オキシム基、１，２，３−トリアゾール基、カルボニル基又はエステル基を有するリンカーであり、Peptideは、下記式で表されるアミノ酸配列：
−ＮＨ−Ｘ１−Ｌｅｕ−Ｘ２−Ｌｅｕ−Ｘ３−Ｘ４−Ｘ５−Ｘ６−Ｘ７−ＣＯＮＨ₂、又はＨ₂Ｎ−Ｘ１−Ｌｅｕ−Ｘ２−Ｌｅｕ−Ｘ３−X４−Ｘ５−Ｘ６−Ｘ７−ＣＯ−（式中、Ｘ１はＡｒｇ又はＡｌａであり、Ｘ２はＴｙｒ、Ａｒｇ又はＬｅｕであり、Ｘ３はＡｒｇ又はＡｌａであり、X４はＩｌｅ、Ｌｅｕ又はＶａｌであり、Ｘ５はＧｌｙ又はＡｒｇであり、Ｘ６はＡｒｇ又はＬｙｓであり、Ｘ７はＡｒｇ又はＬｙｓである。ここで、Ａｒｇ−ＣＯＮＨ₂又はＬｙｓ−ＣＯＮＨ₂は、Ａｒｇ又はＬｙｓのカルボキシル基がアミド化していることを示す。）より選ばれる抗菌活性を有するペプチドである。）
で表される反復単位を含む抗菌性ペプチド含有ポリマー。

Wherein R ₁ is a hydrogen atom or a methyl group, U ₁ is a single bond, O or NH, V is a single bond or a spacer, W is a single bond or an amide group, a thioether group, A linker having a thioester group, an oxime group, a 1,2,3-triazole group, a carbonyl group or an ester group, and Peptide is an amino acid sequence represented by the following formula:
-NH-X1-Leu-X2- Leu-X3-X4-X5-X6-X7-CONH 2, or _{H 2 N-X1-Leu-} X2-Leu-X3-X4-X5-X6-X7-CO- ( In the formula, X1 is Arg or Ala, X2 is Tyr, Arg or Leu, X3 is Arg or Ala, X4 is Ile, Leu or Val, X5 is Gly or Arg, and X6 is Arg Or Lys and X7 is Arg or Lys, where Arg-CONH ₂ or Lys-CONH ₂ indicates that the carboxyl group of Arg or Lys is amidated.) It is a peptide. )
The antibacterial peptide containing polymer containing the repeating unit represented by these.

(2) Further, the following formula (2) and the following formula (3)

（式中Ｒ₂及びＲ₃は、それぞれ独立に水素原子又はメチル基であり、Ｕ₂は、単結合、Ｏ又はＮＨであり、Ｕ₃は、Ｏ又はＮＨであり、Ｖは、単結合又はスペーサーであり、Ｗ₁は、水素原子又は末端にハロゲンを付加してもよい上記リンカーＷの全部又は一部からなる基であり、Ｙは、単結合又は分子量１０〜３０００の置換基を有してもよい直鎖又は分岐状のアルキレン鎖又はポリオキシアルキレン鎖である。）
で表される反復単位のうちの少なくとも一つを含む上記（１）に記載の抗菌性ペプチド含有ポリマー。
（３）前記式（１）〜（３）において、−Ｗ−が、下記式：

(Wherein R ₂ and R ₃ are each independently a hydrogen atom or a methyl group, U ₂ is a single bond, O or NH, U ₃ is O or NH, and V is a single bond or A spacer, W ₁ is a hydrogen atom or a group consisting of all or part of the linker W to which a halogen may be added at the end, and Y has a single bond or a substituent having a molecular weight of 10 to 3000 It may be a linear or branched alkylene chain or polyoxyalkylene chain.)
The antibacterial peptide-containing polymer according to (1), which comprises at least one of the repeating units represented by
(3) In the formulas (1) to (3), -W- represents the following formula:

（式中、ｔは、１〜７の自然数である。）のいずれかを有するリンカーであり、かつ、Ｙが、単結合又は分子量１０〜３０００の置換基を有してもよい直鎖又は分岐状のアルキレン鎖又はポリエチレングリコール鎖である、上記（１）または（２）に記載の抗菌性ペプチド含有ポリマー。
（４）前記式（１）の−Ｗ−Peptideが、以下に表される式（１１）〜（２７）：

(Wherein t is a natural number of 1 to 7), and Y may have a single bond or a substituent having a molecular weight of 10 to 3000, linear or branched The antibacterial peptide-containing polymer according to the above (1) or (2), which is an alkylene chain or a polyethylene glycol chain.
(4) -W-Peptide in the formula (1) is represented by the following formulas (11) to (27):

（ここで、Peptideは前記と同じである。），

(Where Peptide is the same as above),

（ここで、Peptideは前記と同じであり、ｉは１〜１５の整数である。），

(Here, Peptide is the same as described above, and i is an integer of 1 to 15.),

（ここで、Peptideは前記と同じであり、ｉは１〜７の整数である。），

(Here, Peptide is the same as described above, and i is an integer of 1 to 7.),

（ここで、Peptideは前記と同じであり、ｉは、１〜７の整数であり、ｊは、１〜１５の整数である。），

(Here, Peptide is the same as described above, i is an integer of 1 to 7, and j is an integer of 1 to 15.),

（ここで、Peptideは前記と同じである。），

(Where Peptide is the same as above),

（ここで、Peptideは前記と同じであり、ｉは、１〜１５の整数である。），

(Here, Peptide is the same as described above, and i is an integer of 1 to 15.),

（ここで、Peptideは前記と同じであり、ｉは１〜８の整数であり、ｊは１〜８の整数である。），

(Here, Peptide is the same as above, i is an integer of 1 to 8, and j is an integer of 1 to 8.),

（ここで、Peptideは前記と同じであり、ｉは１〜７の整数であり、ｊは１〜７の整数である。），

(Here, Peptide is the same as described above, i is an integer of 1 to 7, and j is an integer of 1 to 7.),

(ここで、Peptideは前記と同じである。），

(Where Peptide is the same as above),

（ここで、Peptideは前記と同じであり、ｉは２〜７の整数である。），

(Here, Peptide is the same as described above, and i is an integer of 2 to 7.),

（ここで、Peptideは前記と同じであり、i は１〜７の整数であり、j は２〜７の整数である。），

(Wherein Peptide is the same as above, i is an integer of 1 to 7, and j is an integer of 2 to 7),

（ここで、Peptideは前記と同じであり、i は１〜１５の整数であり、j は１〜１０の整数である。），

(Here, Peptide is the same as above, i is an integer of 1 to 15, and j is an integer of 1 to 10.),

（ここで、Peptideは前記と同じであり、i は１〜８の整数であり、j は１〜７の整数である。），

(Where Peptide is the same as above, i is an integer of 1 to 8, and j is an integer of 1 to 7),

（ここで、Peptideは前記と同じであり、i は１〜７の整数であり、j は１〜７の整数である。），

(Where Peptide is the same as above, i is an integer of 1 to 7, and j is an integer of 1 to 7),

（ここで、Peptideは前記と同じであり、ｉは２〜７の整数である。）

(Here, Peptide is the same as described above, and i is an integer of 2 to 7.)

（ここで、Peptideは前記と同じであり、i は１〜１０の整数であり、j は１〜１５の整数である。）,

(Here, Peptide is the same as described above, i is an integer of 1 to 10, and j is an integer of 1 to 15.),

（ここで、Peptideは前記と同じであり、i は２〜７の整数である。）、及び

(Where Peptide is the same as above and i is an integer from 2 to 7), and

（ここで、Peptideは前記と同じであり、i は１〜１０の整数である。）
からなる群から選ばれ、かつ、
前記式（２）の−Ｗ₁が、下記式（３１）〜（３８）：

(Here, Peptide is the same as described above, and i is an integer of 1 to 10.)
Selected from the group consisting of
-W ₁ is a compound represented by the following formula in the formula (2) (31) to (38):

（ここで、ｉは、１〜７の整数であり、ｊは１〜８の整数であり、ｋは１〜１５の整数であり、ｌは０〜８の整数であり、ｈは１〜５の整数であり、Ｘは、Ｃｌ、Ｂｒ又はＩである）
からなる群から選ばれる、前記（２）に記載の抗菌性ペプチド含有ポリマー。

(Where i is an integer from 1 to 7, j is an integer from 1 to 8, k is an integer from 1 to 15, l is an integer from 0 to 8, and h is from 1 to 5) And X is Cl, Br or I)
The antibacterial peptide-containing polymer according to (2), selected from the group consisting of:

In the antimicrobial peptide-containing polymers, preferred combinations of -W ₁ in the formula (1) -W-Peptide in the formula in (2) is a combination shown below.
Formula (11) -Formula (31); Formula (12) -Formula (31); Formula (13) -Formula (32); Formula (14) -Formula (32); Formula (15) -Formula (33); Formula (16) -Formula (33); Formula (17) -Formula (34); Formula (18) -Formula (35); Formula (19) -Formula (33); Formula (20) -Formula (31); Formula (21) -Formula (32); Formula (22) -Formula (36); Formula (23) -Formula (37); Formula (24) -Formula (38); Formula (25) -Formula (39); Formula (26) -Formula (39); Formula (27) -Formula (33); Formula (28) -Formula (33).

(5) -W-Peptide in the formula (1) is represented by the following formula:

（ここで、Peptideは前記と同じである。）であり、前記式（２）のＷ₁が、下記式：

(Where Peptide is the same as above), and W _{1 in the} formula (2) is the following formula:

である前記（２）に記載の抗菌性ペプチド含有ポリマー。

The antibacterial peptide-containing polymer as described in (2) above.

(6) The spacer V may have a linear or branched polyethylene glycol which may have a substituent having a weight average molecular weight of 50 to 1000, or a linear chain which may have a substituent having a molecular weight of 10 to 500. Or the antibacterial peptide containing polymer in any one of said (1)-(5) which is a branched alkylene chain, or those combination.

(7) Following formula (4)

で表される反復単位、及び下記式（５）

And a repeating unit represented by the following formula (5):

で表される反復単位を含む抗菌性ペプチド含有ポリマー。

The antibacterial peptide containing polymer containing the repeating unit represented by these.

(8) The following formulas (1), (2) and (3)

で表される反復単位を含む抗菌性ペプチド含有ポリマーであって、
式（１）中、Ｕ₁がＯで、−Ｖ−Ｗ−Peptideが、−ＰＥＧ−Ｓ−Ｃｙｓ−Peptide（ここで、ＰＥＧは、重量平均分子量が１０〜２０００のポリエチレングリコールである。）又は-ＣＨ₂ＣＨＯＨＣＨ₂−Ｓ-Ｃｙｓ-Peptide、又は、Ｕ₁がＮＨで、−Ｖ−Ｗ−Peptideが、-Peptide、−（ＣＨ₂）₃−Ｓ−Ｃｙｓ−Peptide、−ＣＨ₂−（Ｃ₂Ｈ₄Ｏ）₃−Ｃ₃Ｈ₆−ＮＨ−ＣＯＣＨ₂−Ｓ−Ｃｙｓ−Peptide、及び下記式：

An antibacterial peptide-containing polymer comprising a repeating unit represented by
In the formula (1), U ₁ is O, -VW-Peptide is -PEG-S-Cys-Peptide (wherein PEG is polyethylene glycol having a weight average molecular weight of 10 to 2000) or _{-CH 2 CHOHCH 2 -S-Cys-} Peptide, or, in U ₁ is NH, is -V-W-Peptide, -Peptide, - (CH 2) 3 -S-Cys-Peptide, -CH 2 - (C _{2 H 4 O) 3 -C 3} H 6 -NH-COCH 2 -S-Cys-Peptide, and the following formula:

からなる群より選ばれ、かつ、
式（３）中、Ｕ₃がＯで、−Ｙ−Ｈが、−（ＣＨ₂）₁₁−ＣＨ₃、又は−ＰＥＧ−Ｒ₃（ここで、ＰＥＧは、重量平均分子量が１０〜２０００のポリエチレングリコールであり、Ｒ₃はＯＨ又はＣＨ₃である。）、又はＵ₃がＮＨで、−Ｙが単結合である、請求項２に記載の抗菌性ペプチド含有ポリマー。

Selected from the group consisting of and
In the formula (3), U ₃ is O, -YH is-(CH ₂ ) ₁₁ -CH ₃ , or -PEG-R ₃ (wherein PEG is a polyethylene having a weight average molecular weight of 10 to 2000) a glycol, R ₃ is OH or CH _3.), or U ₃ is in NH, -Y is a single bond, an antimicrobial peptide-containing polymer according to claim 2.

(9) The antibacterial peptide-containing polymer according to any one of (1) to (8), wherein the Peptide-containing unit is contained in the polymer at a ratio of 1 to 100 mol%.
(10) The antibacterial peptide content according to (9), wherein the Peptide-containing unit is contained in the polymer at a ratio of 0.4 to 20 mol% (more preferably 0.4 to 2 mol%). polymer.

(11) The peptide is
-HN-Arg-Leu-Tyr- Leu-Arg-Ile-Gly-Arg-Arg-CONH 2, -NH-Arg-Leu-Arg-Leu-Arg-Ile-Gly-Arg-Arg-CONH 2, -NH -Ala-Leu-Tyr-Leu- Ala-Ile-Arg-Arg-Arg-CONH 2, -HN-Arg-Leu-Leu-Leu-Arg-Ile-Gly-Arg-Arg-CONH 2, -HN-Arg -Leu-Tyr-Leu-Arg- Val-Gly-Arg-Arg-CONH 2, -NH-Arg-Leu-Arg-Leu-Arg-Val-Gly-Arg-Arg-CONH 2, -NH-Ala-Leu -Tyr-Leu-Ala-Val- Arg-Arg-Arg-CONH 2, -HN-Arg-Leu Leu-Leu-Arg-Val- Gly-Arg-Arg-CONH 2, H 2 N-Arg-Leu-Tyr-Leu-Arg-Ile-Gly-Arg-Arg-CO-, H 2 N-Arg-Leu- Arg-Leu-Arg-Ile- Gly-Arg-Arg-CO-, H 2 N-Ala-Leu-Tyr-Leu-Ala-Ile-Arg-Arg-Arg-CO-, H 2 N-Arg-Leu- Leu-Leu-Arg-Ile- Gly-Arg-Arg-CO-, H 2 N-Arg-Leu-Tyr-Leu-Arg-Val-Gly-Arg-Arg-CO-, H 2 N-Arg-Leu- Arg-Leu-Arg-Val- Gly-Arg-Arg-CO-, H 2 N-Ala-Leu-Tyr-Leu-Ala-Val-Arg-Ar -Arg-CO-, and _{H 2 N-Arg-Leu-} Leu-Leu-Arg-Val-Gly-Arg-Arg-CO- wherein a peptide chosen from the group consisting of (1) either - (10) An antibacterial peptide-containing polymer according to claim 1.

(12) An antibacterial fiber in which the antibacterial peptide-containing polymer according to any one of (1) to (11) is coated on a fiber.
(13) The antibacterial fiber according to (12), wherein the antibacterial peptide-containing polymer is present in an amount of 0.001 μmol or more and 0.3 μmol or less per 1 cm ² of the cloth.
(14) The antibacterial fiber according to the above (11) or (12) in medical use or sanitary use.
(15) A method for immobilizing an antimicrobial peptide on a fiber by coating the antimicrobial peptide-containing polymer according to any one of (1) to (11) on the fiber.
(16) A polymer emulsion containing the antibacterial peptide-containing polymer according to any one of (1) to (11).

(17) Formula (9) below

（式中Ｒ₁は、水素原子又はメチル基であり、Ｕ₁は、Ｏ又はＮＨであり、Ｖは、単結合又はスペーサーであり、Ｗoは、ペプチド又は誘導体化したペプチドと反応する官能基である。）
で表される反復単位を含むポリマーに、下記式（１０）：

Wherein R ₁ is a hydrogen atom or a methyl group, U ₁ is O or NH, V is a single bond or a spacer, Wo is a functional group that reacts with a peptide or a derivatized peptide. is there.)
In the polymer containing the repeating unit represented by the following formula (10):

（ここで、Peptideは、下記式で表されるアミノ酸配列：
−ＮＨ−Ｘ１−Ｌｅｕ−Ｘ２−Ｌｅｕ−Ｘ３−X４−Ｘ５−Ｘ６−Ｘ７−ＣＯＮＨ₂、又はＨ₂Ｎ−Ｘ１−Ｌｅｕ−Ｘ２−Ｌｅｕ−Ｘ３−X４−Ｘ５−Ｘ６−Ｘ７−ＣＯ−（式中、Ｘ１はＡｒｇ又はＡｌａであり、Ｘ２はＴｙｒ、Ａｒｇ又はＬｅｕであり、Ｘ３はＡｒｇ又はＡｌａであり、X４はＩｌｅ又はＬｅｕ又はＶａｌであり、Ｘ５はＧｌｙ又はＡｒｇであり、Ｘ６は、Ａｒｇ又はＬｙｓであり、Ｘ７は、Ａｒｇ又はＬｙｓである。ここで、Ａｒｇ−ＣＯＮＨ₂又はＬｙｓ−ＣＯＮＨ₂は、Ａｒｇ又はＬｙｓのカルボキシル基がアミド化していることを示す。）より選ばれる抗菌活性を有するペプチドである。）より選ばれ、Ｚは、水素原子、水酸基、Ｃｙｓ又はＷoと反応しリンカーを形成する官能基（例えば、アジド基、スルフヒドリル基、ハロゲン基、アルキン基、カルボニル基、ヒドロキシルアミノ基）を有する分子量５０〜５００の分子である。）
で表されペプチド又はその誘導体を反応させることにより、抗菌性ペプチド含有ポリマーを作製する方法。

(Here, Peptide is an amino acid sequence represented by the following formula:
-NH-X1-Leu-X2- Leu-X3-X4-X5-X6-X7-CONH 2, or _{H 2 N-X1-Leu-} X2-Leu-X3-X4-X5-X6-X7-CO- ( In the formula, X1 is Arg or Ala, X2 is Tyr, Arg or Leu, X3 is Arg or Ala, X4 is Ile or Leu or Val, X5 is Gly or Arg, and X6 is Arg or Lys and X7 is Arg or Lys, where Arg-CONH ₂ or Lys-CONH ₂ indicates that the carboxyl group of Arg or Lys is amidated.) Is a peptide having Z is a molecular weight having a functional group (for example, azide group, sulfhydryl group, halogen group, alkyne group, carbonyl group, hydroxylamino group) that reacts with a hydrogen atom, a hydroxyl group, Cys or Wo to form a linker. 50 to 500 molecules. )
A method for producing an antibacterial peptide-containing polymer by reacting a peptide or a derivative thereof.

(18) The following formulas (9) and (3)

（式中Ｒ₁及びＲ₃は、それぞれ独立に水素原子又はメチル基であり、Ｕ₁及びＵ₃は、それぞれ独立にＯ又はＮＨであり、Ｖは、単結合又はスペーサーであり、Ｗoは、ペプチド又は誘導体化したペプチドと反応する官能基であり、Ｙは、単結合又は分子量１０〜３０００の置換基を有してもよい直鎖又は分岐状のアルキレン鎖又はポリオキシアルキレン鎖である。）
で表される反復単位を含むポリマーに、下記式（１０）：

Wherein R ₁ and R ₃ are each independently a hydrogen atom or a methyl group, U ₁ and U ₃ are each independently O or NH, V is a single bond or a spacer, Wo is A functional group that reacts with a peptide or a derivatized peptide, and Y is a single bond or a linear or branched alkylene chain or polyoxyalkylene chain that may have a substituent having a molecular weight of 10 to 3000.
In the polymer containing the repeating unit represented by the following formula (10):

（ここで、Peptideは、下記式で表されるアミノ酸配列：
−ＮＨ−Ｘ１−Ｌｅｕ−Ｘ２−Ｌｅｕ−Ｘ３−X４−Ｘ５−Ｘ６−Ｘ７−ＣＯＮＨ₂、又はＨ₂Ｎ−Ｘ１−Ｌｅｕ−Ｘ２−Ｌｅｕ−Ｘ３−X４−Ｘ５−Ｘ６−Ｘ７−ＣＯ−（式中、Ｘ１はＡｒｇ又はＡｌａであり、Ｘ２はＴｙｒ、Ａｒｇ又はＬｅｕであり、Ｘ３はＡｒｇ又はＡｌａであり、X４はＩｌｅ又はＬｅｕ又はＶａｌであり、Ｘ５はＧｌｙ又はＡｒｇであり、Ｘ６は、Ａｒｇ又はＬｙｓであり、Ｘ７は、Ａｒｇ又はＬｙｓである。ここで、Ａｒｇ−ＣＯＮＨ₂又はＬｙｓ−ＣＯＮＨ₂は、Ａｒｇ又はＬｙｓのカルボキシル基がアミド化していることを示す。）より選ばれる抗菌活性を有するペプチドである。）より選ばれ、Ｚは、水素原子、水酸基、Ｃｙｓ又はＷoと反応しリンカーを形成する官能基（例えば、アジド基、スルフヒドリル基、ハロゲン基、アルキン基、カルボニル基、ヒドロキシルアミノ基）を有する分子量５０〜５００の分子である。）
で表されペプチド又はその誘導体を反応させることにより、抗菌性ペプチド含有ポリマーを作製する方法。

(Here, Peptide is an amino acid sequence represented by the following formula:
-NH-X1-Leu-X2- Leu-X3-X4-X5-X6-X7-CONH 2, or _{H 2 N-X1-Leu-} X2-Leu-X3-X4-X5-X6-X7-CO- ( In the formula, X1 is Arg or Ala, X2 is Tyr, Arg or Leu, X3 is Arg or Ala, X4 is Ile or Leu or Val, X5 is Gly or Arg, and X6 is Arg or Lys and X7 is Arg or Lys, where Arg-CONH ₂ or Lys-CONH ₂ indicates that the carboxyl group of Arg or Lys is amidated.) Is a peptide having Z is a molecular weight having a functional group (for example, azide group, sulfhydryl group, halogen group, alkyne group, carbonyl group, hydroxylamino group) that reacts with a hydrogen atom, a hydroxyl group, Cys or Wo to form a linker. 50 to 500 molecules. )
A method for producing an antibacterial peptide-containing polymer by reacting a peptide or a derivative thereof.

(19) 1 The repeating unit represented by the formula (9) is represented by the following formula (6)

（ここで、式中Ｒ₂は、水素原子又はメチル基である。）
であり、かつ、前記ペプチドが、前記式（１０）においてＺがＣｙｓである、前記（１７）又は（１８）に記載の方法。

(Here, R ₂ is a hydrogen atom or a methyl group.)
And the peptide according to (17) or (18), wherein in the formula (10), Z is Cys.

(20) The following steps:
Process (1): following formula (9)

（式中Ｒ₁は、水素原子又はメチル基であり、Ｕ₁は、Ｏ又はＮＨであり、Ｖは、単結合又はスペーサーであり、Ｗoは、ペプチドまたは誘導体化したペプチドと反応する官能基である。）
で表される反復単位を含むポリマーを含む溶液を繊維にコーティングする工程、及び
工程（２）：下記式：

(Wherein R ₁ is a hydrogen atom or a methyl group, U ₁ is O or NH, V is a single bond or a spacer, Wo is a functional group that reacts with a peptide or a derivatized peptide) is there.)
A step of coating a fiber with a solution containing a polymer containing a repeating unit represented by formula (2): and the following formula:

（ここで、Peptideは、下記式で表されるアミノ酸配列：
−ＮＨ−Ｘ１−Ｌｅｕ−Ｘ２−Ｌｅｕ−Ｘ３−X４−Ｘ５−Ｘ６−Ｘ７−ＣＯＮＨ₂、又はＨ₂Ｎ−Ｘ１−Ｌｅｕ−Ｘ２−Ｌｅｕ−Ｘ３−X４−Ｘ５−Ｘ６−Ｘ７−ＣＯ−（式中、Ｘ１はＡｒｇ又はＡｌａであり、Ｘ２はＴｙｒ、Ａｒｇ又はＬｅｕであり、Ｘ３はＡｒｇ又はＡｌａであり、X４はＩｌｅ又はＬｅｕ又はＶａｌであり、Ｘ５はＧｌｙ又はＡｒｇであり、Ｘ６は、Ａｒｇ又はＬｙｓであり、Ｘ７は、Ａｒｇ又はＬｙｓである。ここで、Ａｒｇ−ＣＯＮＨ₂又はＬｙｓ−ＣＯＮＨ₂は、Ａｒｇ又はＬｙｓのカルボキシル基がアミド化していることを示す。）より選ばれる抗菌活性を有するペプチドである。）より選ばれ、Ｚは、水素原子、水酸基、Ｃｙｓ又はＷoと反応しリンカーを形成する官能基（例えば、アジド基、スルフヒドリル基、ハロゲン基、アルキン基、カルボニル基、ヒドロキシルアミノ基）を有する分子量５０〜５００の分子である。）
で表される抗菌性ペプチド又はその誘導体を含む溶液に、前記ポリマー溶液をコーティングした繊維を浸漬する工程、及び
工程（３）：１〜１０時間放置することによりポリマーにペプチドを結合させた後、該繊維を洗浄して未反応のペプチド又はその誘導体を除去するする工程、
からなる抗菌性繊維の製造方法。

(Here, Peptide is an amino acid sequence represented by the following formula:
-NH-X1-Leu-X2- Leu-X3-X4-X5-X6-X7-CONH 2, or _{H 2 N-X1-Leu-} X2-Leu-X3-X4-X5-X6-X7-CO- ( In the formula, X1 is Arg or Ala, X2 is Tyr, Arg or Leu, X3 is Arg or Ala, X4 is Ile or Leu or Val, X5 is Gly or Arg, and X6 is Arg or Lys and X7 is Arg or Lys, where Arg-CONH ₂ or Lys-CONH ₂ indicates that the carboxyl group of Arg or Lys is amidated.) Is a peptide having Z is a molecular weight having a functional group (for example, azide group, sulfhydryl group, halogen group, alkyne group, carbonyl group, hydroxylamino group) that reacts with a hydrogen atom, a hydroxyl group, Cys or Wo to form a linker. 50 to 500 molecules. )
A step of immersing the fiber coated with the polymer solution in a solution containing the antibacterial peptide or derivative thereof represented by: and step (3): after allowing the peptide to bind to the polymer by leaving it for 1 to 10 hours, Washing the fibers to remove unreacted peptides or derivatives thereof;
The manufacturing method of the antimicrobial fiber which consists of.

(21) The following steps:
Step (1): Formulas (9) and (3) below

（式中Ｒ₁及びＲ₃は、それぞれ独立に水素原子又はメチル基であり、Ｕ₁及びＵ₃は、それぞれ独立にＯ又はＮＨであり、Ｖは、単結合又はスペーサーであり、Ｗoは、ペプチド又は誘導体化したペプチドと反応する官能基であり、Ｙは、単結合又は分子量１０〜３０００の置換基を有してもよい直鎖又は分岐状のアルキレン鎖又はポリオキシアルキレン鎖である。）
で表される反復単位を含むポリマーを含む溶液を繊維にコーティングする工程、及び
工程（２）：下記式（１０）：

Wherein R ₁ and R ₃ are each independently a hydrogen atom or a methyl group, U ₁ and U ₃ are each independently O or NH, V is a single bond or a spacer, Wo is A functional group that reacts with a peptide or a derivatized peptide, and Y is a single bond or a linear or branched alkylene chain or polyoxyalkylene chain that may have a substituent having a molecular weight of 10 to 3000.
A step of coating a fiber with a solution containing a polymer containing a repeating unit represented by formula (2): the following formula (10):

（ここで、Peptideは、下記式で表されるアミノ酸配列：
−ＮＨ−Ｘ１−Ｌｅｕ−Ｘ２−Ｌｅｕ−Ｘ３−X４−Ｘ５−Ｘ６−Ｘ７−ＣＯＮＨ₂、又はＨ₂Ｎ−Ｘ１−Ｌｅｕ−Ｘ２−Ｌｅｕ−Ｘ３−X４−Ｘ５−Ｘ６−Ｘ７−ＣＯ−（式中、Ｘ１はＡｒｇ又はＡｌａであり、Ｘ２はＴｙｒ、Ａｒｇ又はＬｅｕであり、Ｘ３はＡｒｇ又はＡｌａであり、X４はＩｌｅ又はＬｅｕ又はＶａｌであり、Ｘ５はＧｌｙ又はＡｒｇであり、Ｘ６は、Ａｒｇ又はＬｙｓであり、Ｘ７は、Ａｒｇ又はＬｙｓである。ここで、Ａｒｇ−ＣＯＮＨ₂又はＬｙｓ−ＣＯＮＨ₂は、Ａｒｇ又はＬｙｓのカルボキシル基がアミド化していることを示す。）より選ばれる抗菌活性を有するペプチドである。）より選ばれ、Ｚは、水素原子、水酸基、Ｃｙｓ又はＷoと反応しリンカーを形成する官能基（例えば、アジド基、スルフヒドリル基、ハロゲン基、アルキン基、カルボニル基、ヒドロキシルアミノ基）を有する分子量５０〜５００の分子である。）
で表される抗菌性ペプチド又はその誘導体を含む溶液に、前記ポリマー溶液をコーティングした繊維を浸漬する工程、及び
工程（３）：１〜１０時間放置することによりポリマーにペプチドを結合させた後、該繊維を洗浄して未反応のペプチド又はその誘導体を除去するする工程、
からなる抗菌性繊維の製造方法。

(Here, Peptide is an amino acid sequence represented by the following formula:
-NH-X1-Leu-X2- Leu-X3-X4-X5-X6-X7-CONH 2, or _{H 2 N-X1-Leu-} X2-Leu-X3-X4-X5-X6-X7-CO- ( In the formula, X1 is Arg or Ala, X2 is Tyr, Arg or Leu, X3 is Arg or Ala, X4 is Ile or Leu or Val, X5 is Gly or Arg, and X6 is Arg or Lys and X7 is Arg or Lys, where Arg-CONH ₂ or Lys-CONH ₂ indicates that the carboxyl group of Arg or Lys is amidated.) Is a peptide having Z is a molecular weight having a functional group (for example, azide group, sulfhydryl group, halogen group, alkyne group, carbonyl group, hydroxylamino group) that reacts with a hydrogen atom, a hydroxyl group, Cys or Wo to form a linker. 50 to 500 molecules. )
A step of immersing the fiber coated with the polymer solution in a solution containing the antibacterial peptide or derivative thereof represented by: and step (3): after allowing the peptide to bind to the polymer by leaving it for 1 to 10 hours, Washing the fibers to remove unreacted peptides or derivatives thereof;
The manufacturing method of the antimicrobial fiber which consists of.

(22) The repeating unit represented by the formula (9) is represented by the following formula (6).

（ここで、式中Ｒ₂は、水素原子又はメチル基である。）
であり、かつ、前記ペプチドが、前記式（１０）においてＺがＣｙｓである、前記（２０）又は（２１）に記載の抗菌性繊維の製造方法。

(Here, R ₂ is a hydrogen atom or a methyl group.)
The method for producing an antibacterial fiber according to (20) or (21), wherein the peptide is Z in the formula (10) and Cys.

The antimicrobial peptide-containing polymer of the present invention can also be expressed as follows.
The following formula (1a):

（式中ｎは自然数であり、ｍ１は０又は自然数であり、Ｒ₁、R₂、Ｕ₁、Ｕ₂、Ｖ、Ｗ、Ｗ₁、Peptideは、前記と同じである）で表される抗菌性ペプチド含有ポリマー、又は、
下記式（２ａ）：

Wherein n is a natural number, m1 is 0 or a natural number, and R ₁ , R ₂ , U ₁ , U ₂ , V, W, W ₁ , and Peptide are the same as described above. A peptide-containing polymer, or
The following formula (2a):

（式中ｎ及びｍ２は自然数であり、Ｒ₁、R₃、Ｕ₁、Ｕ₃、Ｖ、Ｗ、Ｙ、Peptideは、前記と同じである。）で表される抗菌性ペプチド含有ポリマー、又は、
下記式（３ａ）：

Wherein n and m2 are natural numbers, and R ₁ , R ₃ , U ₁ , U ₃ , V, W, Y, and Peptide are the same as described above, or ,
The following formula (3a):

（式中ｎ、ｍ１及びｍ２は自然数であり、Ｒ₁、R₂、R₃、Ｕ₁、Ｕ₂、Ｕ₃、Ｖ、Ｗ、Ｗ₁Ｙ、及びPeptideは、前記と同じである。）で表される抗菌性ペプチド含有ポリマー。

(In the formula, n, m1 and m2 are natural numbers, and R ₁ , R ₂ , R ₃ , U ₁ , U ₂ , U ₃ , V, W, W ₁ Y and Peptide are the same as described above.) An antibacterial peptide-containing polymer represented by

The antimicrobial peptide-containing polymer of the present invention can also be expressed as follows.
Following formula (4a):

（ここで、ｎは自然数であり、ｍ１は０又は自然数であり、Peptideは前記と同じである。）で表される抗菌性ペプチド含有ポリマー。

(Where n is a natural number, m1 is 0 or a natural number, and Peptide is the same as described above).

The method for producing the antibacterial peptide-containing polymer of the present invention can also be expressed as follows.
Step (1); the following formula (1b):

（式中、ｎは自然数であり、ｍ2は０又は自然数であり、Ｒ₁、Ｒ₃、Ｕ₁、Ｕ₃、Ｖ、Ｗo、Ｙは、上記と同じである。）
で表されるポリマーに、下記式（１０）：

(In the formula, n is a natural number, m2 is 0 or a natural number, and R ₁ , R ₃ , U ₁ , U ₃ , V, Wo, and Y are the same as above.)
In the polymer represented by the following formula (10):

（式中、Peptideは、上記と同じであり、Ｚは、水素原子、水酸基、Ｃｙｓ又はＷoと反応しリンカーを形成する官能基（例えば、アジド基、スルフヒドリル基、ハロゲン基、アルキン基、カルボニル基、ヒドロキシルアミノ基）を有する分子量５０〜５００の分子である。）
で表されるペプチド又はその誘導体を反応させることにより、抗菌性ペプチド含有ポリマーを作製する方法。

(Wherein Peptide is the same as above, and Z is a functional group that reacts with a hydrogen atom, a hydroxyl group, Cys or Wo to form a linker (for example, an azide group, a sulfhydryl group, a halogen group, an alkyne group, a carbonyl group). A molecule having a molecular weight of 50 to 500 having a hydroxylamino group).
A method for producing an antibacterial peptide-containing polymer by reacting the peptide represented by

The method for producing the antibacterial fiber of the present invention can also be expressed as follows.
The following formula (1b):

（式中、ｎは自然数であり、ｍ2は０又は自然数であり、Ｒ₁、Ｒ₃、Ｕ₁、Ｕ₃、Ｖ、Ｗo、Ｙは、上記と同じである。）
で表されるポリマーを含む溶液を繊維にコーティングする工程、及び
工程（２）；下記式（１０）：

(In the formula, n is a natural number, m2 is 0 or a natural number, and R ₁ , R ₃ , U ₁ , U ₃ , V, Wo, and Y are the same as above.)
A step of coating a fiber with a solution containing a polymer represented by formula (2); and the following formula (10):

（ここで、Peptideは、上記と同じであり、Ｚは、水素原子、水酸基、Ｃｙｓ又はＷoと反応しリンカーを形成する官能基（例えば、アジド基、スルフヒドリル基、ハロゲン基、アルキン基、カルボニル基、ヒドロキシルアミノ基）を有する分子量５０〜５００の分子である。）
で表される抗菌性ペプチド又はその誘導体を含む溶液に、前記ポリマーをコーティングした繊維を浸漬する工程、及び
工程（３）；１〜１０時間放置することによりポリマーにペプチドを結合させた後、該繊維を洗浄して未反応のペプチド又はその誘導体を除去するする工程、
からなる抗菌性繊維の製造方法。

(Here, Peptide is the same as above, and Z is a functional group that reacts with a hydrogen atom, hydroxyl group, Cys or Wo to form a linker (for example, azide group, sulfhydryl group, halogen group, alkyne group, carbonyl group). A molecule having a molecular weight of 50 to 500 having a hydroxylamino group).
The step of immersing the fiber coated with the polymer in a solution containing the antibacterial peptide represented by the following or a derivative thereof; and step (3): after allowing the peptide to bind to the polymer by leaving it for 1 to 10 hours, Washing the fibers to remove unreacted peptides or derivatives thereof;
The manufacturing method of the antimicrobial fiber which consists of.

According to one embodiment of the present invention, an antibacterial peptide (modified beetle defensin-derived modified peptide) -containing polymer that can be used in the production of an antibacterial fiber can be provided.
According to another aspect of the present invention, a fiber in which a peptide is immobilized while retaining its activity can be provided by a relatively easy process using an antibacterial peptide-containing polymer.
According to another embodiment of the present invention, a fiber on which a peptide, particularly an antibacterial peptide is immobilized, can be provided by a relatively easy peptide immobilization method.
The fiber provided in this way can maintain the activity of the peptide. In particular, when an antibacterial peptide is used, the provided fiber has antibacterial activity and is subjected to repeated washing and / or sterilization by autoclaving. However, antibacterial activity can be maintained.
Since the fiber of the present invention is also excellent in safety, it can be used for medical purposes such as medical bandages, sheets, clothes and the like.

In the present specification, the expression “A to B” means that A is included and is A or more, and B is included and is B or less.
In this specification, unless otherwise specified, “fiber” includes any of fibers as a raw material for yarn or fabric, yarn made from the fiber, fabric made from the yarn, or nonwoven fabric made from the fiber. As used herein, the term “coats a polymer on a fiber” means a fiber as a raw material for yarn or fabric. On the other hand, the term “fiber coated with polymer” refers to either a fiber coated with a polymer or a fiber as a raw material of a fabric, a yarn made of a fiber coated with the polymer, or a fabric made of the yarn. Including.
In the present specification, a polypeptide and a peptide are used in the same meaning unless otherwise specified, and can be changed from each other.
In this specification, when it represents with -S-Cys-Peptide, S means S in cysteine.
In the present specification, when a peptide is represented, for example, Leu means a leucine residue.
In the present specification, for example, when a polymer having A and B in the polymer at random is represented, it can be represented by any of the following modes.
Following formula:

及び下記式：

の繰り返し単位を有するポリマー、又は
下記式：

And the following formula:

Or a polymer having a repeating unit of the following formula:

（ここで、ｎおよびｍは自然数である。）
で表されるポリマー。

(Here, n and m are natural numbers.)
A polymer represented by

  In one embodiment, the present invention provides an antibacterial peptide-containing polymer represented by the following formula. The following formula (1a):

（式中Ｒ₁及びR₂は、それぞれ独立に水素原子又はメチル基であり、ｎは自然数であり、ｍ１は０又は自然数であり、Ｕ₁及びＵ₂は、それぞれ独立にＯ又はＮＨであり、Ｖは、単結合又はスペーサーであり、Ｗは、単結合或いは、アミド基、チオエーテル基、チオエステル基、オキシム基、１，２，３−トリアゾール基又はエステル基を有するリンカーであり、Ｗ₁は、水素原子又は末端にハロゲンを付加してもよいリンカーＷの全部又は一部からなる基であり、Peptideは、下記式で表されるアミノ酸配列：−ＮＨ−Ｘ１−Ｌｅｕ−Ｘ２−Ｌｅｕ−Ｘ３−X４−Ｘ５−Ｘ６−Ｘ７−ＣＯＮＨ₂、又はＨ₂Ｎ−Ｘ１−Ｌｅｕ−Ｘ２−Ｌｅｕ−Ｘ３−X４−Ｘ５−Ｘ６−Ｘ７−ＣＯ−（式中、Ｘ１はＡｒｇ又はＡｌａであり、Ｘ２はＴｙｒ、Ａｒｇ又はＬｅｕであり、Ｘ３はＡｒｇ又はＡｌａであり、X４はＩｌｅ又はＬｅｕ又はＶａｌであり、Ｘ５はＧｌｙ又はＡｒｇであり、Ｘ６は、Ａｒｇ又はＬｙｓであり、Ｘ７は、Ａｒｇ又はＬｙｓである。ここで、Ａｒｇ−ＣＯＮＨ₂又はＬｙｓ−ＣＯＮＨ₂は、Ａｒｇ又はＬｙｓのカルボキシル基がアミド化していることを示す。）より選ばれる抗菌活性を有するポリペプチドである。）で表される抗菌性ペプチド含有ポリマー、下記式（２ａ）：

Wherein R ₁ and R ₂ are each independently a hydrogen atom or a methyl group, n is a natural number, m1 is 0 or a natural number, and U ₁ and U ₂ are each independently O or NH. , V is a single bond or a spacer, W is a single bond or a linker having an amide group, a thioether group, a thioester group, an oxime group, a 1,2,3-triazole group or an ester group, and W ₁ is , A hydrogen atom or a group consisting of all or part of a linker W to which a halogen may be added to the terminal, and Peptide is an amino acid sequence represented by the following formula: -NH-X1-Leu-X2-Leu-X3 -X4-X5-X6-X7- CONH 2, or H in _{2 N-X1-Leu-X2} -Leu-X3-X4-X5-X6-X7-CO- ( wherein, X1 is Arg or Ala, X2 Is Tyr, Arg or Leu, X3 is Arg or Ala, X4 is Ile or Leu or Val, X5 is Gly or Arg, X6 is Arg or Lys, and X7 is Arg or Lys. , Arg-CONH ₂ or Lys-CONH ₂ indicates that the carboxyl group of Arg or Lys is amidated.) Is an antimicrobial peptide-containing polymer represented by The following formula (2a):

（式中Ｒ₁及びR₃は、それぞれ独立に水素原子又はメチル基であり、ｎは自然数であり、ｍ２は自然数であり、Ｕ₁は、単結合、Ｏ又はＮＨであり、Ｕ₃は、Ｏ又はＮＨであり、Ｖ、Ｗ、及びPeptideは、上記と同じであり、Ｙは、水素原子又は分子量１０〜３０００の置換基を有してもよい直鎖又は分岐状のアルキレン鎖又はポリオキシアルキレン鎖である。）で表される抗菌性ペプチド含有ポリマー、又は下記式（３ａ）：

(Wherein R ₁ and R ₃ are each independently a hydrogen atom or a methyl group, n is a natural number, m2 is a natural number, U ₁ is a single bond, O or NH, and U ₃ is O or NH, V, W, and Peptide are the same as above, and Y is a hydrogen atom or a linear or branched alkylene chain or polyoxy group that may have a substituent having a molecular weight of 10 to 3000 An antibacterial peptide-containing polymer represented by the following formula (3a):

（式中Ｒ₁、Ｒ₂及びR₃は、それぞれ独立に水素原子又はメチル基であり、ｎ、ｍ１、及びｍ２は自然数であり、Ｕ₁及びＵ₂は、それぞれ独立に単結合、Ｏ又はＮＨであり、Ｕ₃は、Ｏ又はＮＨでありＶ、Ｗ、Peptide、及びＹは、上記と同じである。）
で表される抗菌性ペプチド含有ポリマー。

Wherein R ₁ , R ₂ and R ₃ are each independently a hydrogen atom or a methyl group, n, m1 and m2 are natural numbers, and U ₁ and U ₂ are each independently a single bond, O or NH, U ₃ is O or NH, and V, W, Peptide, and Y are the same as above.)
An antibacterial peptide-containing polymer represented by

The antibacterial peptide-containing polymer of the present invention may be a homopolymer or a copolymer. Peptide may be bonded to all monomer units or to some monomer units. The ratio of the unit to which Peptide is bonded in the polymer (referred to above as RC (CH ₂ ) -CO-U ₁ -VW-Peptide and referred to as “Peptide-containing unit” in this specification) is determined by coating the polymer on the cloth. (Coating) It is not particularly limited as long as the desired amount of peptide can be applied, and is 0.1 to 100 mol%, preferably 0.4 to 20 mol%, more preferably 0.4 to 2 mol. %. By adjusting the ratio of peptide-containing units in the polymer, the amount of polymer applied (coating) to the fabric can be controlled, and homogeneous and suitable antibacterial fibers having the desired antibacterial properties can be easily produced. A unit in which Peptide in the polymer is not bonded (meaning the above-mentioned RC (CH ₂ ) —CO—U ₂ —VW ₁ and / or RC (CH ₂ ) —CO—U ₃ —Y, Appropriate physical properties such as viscosity and solubility can be obtained by appropriately selecting the ratio of “Peptide-free unit”. In particular, in the copolymer, the desired physical properties can be imparted to the polymer by appropriately selecting the Peptide-free unit (RC (CH ₂ ) —CO—U ₃ —Y).
Y is a single bond or a linear or branched alkylene chain or polyoxyalkylene chain which may have a substituent having a molecular weight of 10 to 3000, preferably 100 to 2000. The physical properties of the polymer can be changed by appropriately selecting the length and type of Y.

The peptide constituting the peptide is selected from a modified beetle defensin-derived modified peptide consisting of an amino acid sequence represented by the following formula, and the peptide can be bound to a linker at either the N-terminus or C-terminus.
_{H 2 N-X1-Leu-} X2-Leu-X3-X4-X5-X6-X7-CONH 2 ( wherein, X1 is Arg or Ala, X2 is Tyr, is Arg or Leu, X3 is Arg or Ala, X4 is Ile or Leu or Val, X5 is Gly or Arg, X6 is Arg or Lys, and X7 is Arg or Lys, where Arg-CONH ₂ or Lys- CONH ₂ indicates that the carboxyl group of Arg or Lys is amidated.)

Particularly preferably, the peptide is H ₂ N-Arg-Leu-Tyr-Leu-Arg-Ile-Gly-Arg-Arg-CONH ₂ , H ₂ N-Arg-Leu-Arg-Leu-Arg-Ile-Gly _{-Arg-Arg-CONH 2, H} 2 N-Ala-Leu-Tyr-Leu-Ala-Ile-Arg-Arg-Arg-CONH 2, H 2 N-Arg-Leu-Leu-Leu-Arg-Ile-Gly _{-Arg-Arg-CONH 2, H} 2 N-Arg-Leu-Tyr-Leu-Arg-Val-Gly-Arg-Arg-CO 2, H 2 N-Arg-Leu-Arg-Leu-Arg-Val-Gly _{-Arg-Arg-CO 2, H} 2 N-Ala-Leu-Tyr-Leu-Ala-Val-Arg-Arg-Arg-CO 2 and Selected from _{2 N-Arg-Leu-Leu} -Leu-Arg-Val-Gly-Arg-Arg-CONH 2 Kararanaru group, particularly _{preferably, H 2 N-Arg-Leu} -Leu-Leu-Arg-Val it is a _{-Gly-Arg-Arg-CONH 2} .

  Modified peptides derived from beetle defensin that can be used as an antibacterial peptide in the present invention include, for example, Japanese Patent No. 3273314, Japanese Patent Application Laid-Open No. 2010-184022, and Ishibashi et al., Eur. J. Biochem, 1999 Dec; 266 ( 2): 616-23, which are hereby incorporated by reference.

  Peptide synthesis can be performed using any conventional peptide synthesis method (solid phase synthesis and / or liquid phase synthesis). In addition, when the polymer and the peptide are bonded via Cys, it is possible to introduce Cys at the C-terminal or N-terminal of the synthetic polypeptide in peptide synthesis, which is convenient.

  W in the above formula (1a) is a linker for immobilizing the peptide. The linker is used to connect the N-terminal or C-terminal of the peptide to the spacer or directly to the side chain terminal of the polymer chain, and can be introduced at the N-terminal or C-terminal of the peptide or at the peptide binding site of the spacer. It is also possible to do. Examples of the linker include a compound having a thioether group, a thioester group, an oxime group, a 1,2,3-triazole group or an ester group, or a group thereof, preferably a compound having an amide group or a thioether group. Or a group thereof, but is not limited thereto. In the present specification, a peptide in which a peptide is modified for the purpose of the present invention, such as a peptide into which a linker or a part thereof is introduced, is referred to as a peptide derivative.

When a linker is introduced at one of the peptide end or spacer end, a linker having reactivity with the functional group at the peptide end or spacer end to which the linker is not introduced is introduced at the peptide end or spacer end where the linker is introduced, and bonded. The peptide can be bound to the spacer by carrying out the reaction.
When a linker is introduced into both the peptide end and the spacer end, a part constituting the linker (linker part W _X ) is introduced on one side and the other part constituting the linker (linker part W _Y ) is provided on the other side. By introducing and reacting both linker parts, the peptide can be bound to the spacer. As a result, a linker W is made from W _x and W _y .
When no spacer is used in the above, a linker or linker part can be introduced at the side chain end of the polymer chain, and the peptide can be directly bonded to the side chain end of the polymer chain.

When a functional group having reactivity with the N-terminal amino group or the C-terminal carboxyl group of the peptide is used for bonding, the linker is an amide group or an ester group. The functional group to be a linker can be introduced at the end of the spacer, but a spacer having it at the end can also be used.
The linker site can be introduced into either the N-terminus or C-terminus of the peptide or the spacer terminus to serve as a linker. However, the linker site can be introduced into both the peptide terminus and the spacer terminus, It can also react to serve as a linker. For example, a thiol can be introduced at either the peptide end or the spacer end, a maleimide group can be introduced into the other, and the peptide can be bound to the spacer via a linker having a thioether group by the reaction of the thiol and the maleimide group. .

When the N-terminal of the peptide is bonded to a spacer, the amino group at the N-terminal can be bonded to the spacer by reacting with a spacer having a functional group such as a carboxyl group, a halogen, an aldehyde group, or an isocyanate group. The reaction between the amino group and these functional groups can be performed by a conventional method, and examples thereof include a condensation reaction and an addition reaction.
The peptide is bound to the N-terminal spacer by introducing a thiol group such as cysteine or thioglycolic acid as a linker site at the N-terminus, and reacting with the SH group such as maleimide group, halogen, thioester group, bromoacetyl group, etc. It can also be reacted with a spacer having a functional group to bind to the spacer. Thereby, a peptide and a spacer are couple | bonded by the linker containing a thioether group. The reaction between the SH group and these functional groups can be performed by a conventional method. For example, the reaction with the maleimide group can be performed by a Michael addition reaction. The reaction with halogen can be performed by nucleophilic substitution reaction, and the reaction with thioester is by transesterification.

Alternatively, propiolic acid, 3-butynoic acid or the like having an alkyne as a linker site at the N-terminus of the peptide can be introduced and reacted with a spacer having an azide group that reacts with alkyne to bind to the spacer. The reaction between the alkyne and the azide group can be performed by a conventional method, for example, by a Huisgen reaction.
In addition, 3-oxyaminopropionic acid, 4-oxyaminobutyric acid or the like having an alkoxyamino group as a linker site at the N-terminus of the peptide is introduced, and reacted with a spacer having a ketone that reacts with the alkoxyamino group to bind to the spacer You can also The reaction between the alkoxyamino group and the ketone can be performed by a conventional method, for example, by an oximation reaction.
In addition, these linker sites can be joined by introducing a spacer and a peptide site in reverse.

When the C-terminal of the peptide is bonded to the spacer, the C-terminal carboxyl group can be bonded to the spacer by reacting with a spacer having a functional group such as an amino group or a hydroxyl group. The reaction between the carboxyl group and these functional groups can be carried out by a conventional method, and examples thereof include a condensation reaction using a condensing agent.
Binding to the C-terminal spacer of the peptide is carried out by introducing a thiol group such as cysteine or 2-amino-1-ethanethiol as a linker site at the C-terminus, and reacting with the SH group, a maleimide group, a halogen, a thioester group, It can also be reacted with a spacer having a functional group such as an aldehyde group or a bromoacetyl group to bind to the spacer. The reaction between the SH group and these functional groups can be performed by the conventional method as described above.

  The spacer in the above formulas (1a) to (3a) is not particularly limited as long as it can bind the polymer and the peptide. For example, the alkylene chain, the polyethylene glycol chain, the sugar chain, the polypeptide, or the structure thereof Among them, those containing two or more are preferable, but those having an alkylene chain or polyethylene glycol chain as the main component are preferred. The alkylene chain or the polyethylene glycol chain can be either linear or branched and may have a substituent. The alkylene chain has a molecular weight of preferably 10 to 500, more preferably 20 to 200. The polyethylene glycol chain has a weight average molecular weight of preferably 50 to 1000, more preferably 100 to 500, and still more preferably 100 to 300. Moreover, as long as it does not inhibit the other coupling | bonding reaction, for example, the coupling | bonding of a polymer and a spacer, a spacer and a linker, or a linker and a peptide, you may have a functional group.

  The spacer can be bonded to the polymer by synthesizing a polymer using a compound having a terminal functional group having reactivity with the carboxyl group of the monomer unit and then introducing it by reacting with the carboxyl group of the polymer, or the monomer. This can be achieved by polymerization after introduction at this point. The bond between the carboxyl group of the polymer and the functional group of the spacer is not particularly limited, but the functional group having reactivity with the carboxyl group of the polymer is preferably an amino group or a hydroxyl group. The spacer that binds to the polymer may have a linker moiety (functional group) for binding to the peptide at the end opposite to the binding end to the polymer, as long as the binding to the polymer is not affected.

Specific examples of preferable spacers are listed below, but are not limited thereto.
Spacer A:

スペーサーＢ：

Spacer B:

スペーサーＣ：

Spacer C:

（式中、Ｘは、Ｃｌ又はＢｒである。）

(In the formula, X is Cl or Br.)

  Hereinafter, although the specific example of the linker (linker part) introduce | transduced into the spacer side or the peptide side and its introduction method is shown, it is not limited to these.

1. Spacer side linker parts
[Ls-A]:

リンカー導入法：スペーサー部の脱保護を行った後、スクシンイミジル型のリンカーを導入。
[Ls-B]

Linker introduction method: After deprotecting the spacer part, a succinimidyl type linker is introduced.
[Ls-B]

（式中、Ｘは、Ｃｌ又はＢｒある。）
リンカー導入法：モノマー時点で、スペーサー部の脱保護を行い、縮合により導入後、ポリマー化する。
[Ls-C]

(Wherein X is Cl or Br)
Linker introduction method: At the time of monomer, the spacer part is deprotected, introduced by condensation, and then polymerized.
[Ls-C]

リンカー導入法：モノマー時点で、スペーサー部の脱保護を行い、縮合により導入後、ポリマー化する。

Linker introduction method: At the time of monomer, the spacer part is deprotected, introduced by condensation, and then polymerized.

[Ls-D]

リンカー導入法：モノマー時点で、スペーサー部の脱保護を行い、ヒドロキシアミノ基Boc保護体を縮合により導入後、ポリマー化する。その後ヒドロキシアミノ基の脱保護をおこなう。
[Ls-E]

Linker introduction method: At the time of the monomer, the spacer part is deprotected, and a hydroxyamino group Boc protector is introduced by condensation and then polymerized. Thereafter, the hydroxyamino group is deprotected.
[Ls-E]

リンカー導入法：ポリマー時点で、スペーサー部の脱保護を行い、ＳＨ基Ｔｒｔ保護体を縮合により導入後、ＳＨ基の脱保護をおこなう。
[Ls-F]

Linker introduction method: At the time of polymer, the spacer part is deprotected, and after the SH group Trt protector is introduced by condensation, the SH group is deprotected.
[Ls-F]

リンカー導入法：ポリマー時点でスペーサー部の脱保護を行った後、縮合で導入。

Linker introduction method: After deprotecting the spacer part at the time of polymer, it is introduced by condensation.

[Ls-G]

リンカー導入法：モノマー時点で、スペーサー部の脱保護を行い、縮合により導入後、ポリマー化する。

Linker introduction method: At the time of monomer, the spacer part is deprotected, introduced by condensation, and then polymerized.

2. Peptide side linker parts
[Lp-A]
Cysteine:
Linker introduction method: Introduced into peptide N-terminal or C-terminal during peptide production.
[Lp-B]

リンカー導入法：ペプチドＮ末に導入できる。ＳＨ基保護カルボン酸を保護ペプチドに導入後脱保護する。
[Lp-C]

Linker introduction method: can be introduced into the peptide N-terminal. The SH group-protected carboxylic acid is introduced into the protected peptide and then deprotected.
[Lp-C]

リンカー導入法：ペプチドＮ末端に導入できる。保護ペプチドに導入後脱保護する。

Linker introduction method: can be introduced at the N-terminus of the peptide. Deprotection after introduction into the protected peptide.

[Lp-D]

リンカー導入法：ペプチドＮ末端に導入できる。保護ペプチドに導入後脱保護する。
[Lp-E]

Linker introduction method: can be introduced at the N-terminus of the peptide. Deprotection after introduction into the protected peptide.
[Lp-E]

リンカー導入法：ペプチドＮ末端に導入できる。保護ペプチドにスクシンイミジル型のリンカーを導入し脱保護する。
[Lp-Ｆ]

Linker introduction method: can be introduced at the N-terminus of the peptide. A succinimidyl-type linker is introduced into the protected peptide for deprotection.
[Lp-F]

（式中、Ｘは、Ｃｌ又はＢｒある。）
リンカー導入法：ペプチドのＮ末端に導入できる。保護ペプチドに導入後脱保護する。

(Wherein X is Cl or Br)
Linker introduction method: can be introduced at the N-terminus of the peptide. Deprotection after introduction into the protected peptide.

[Lp-G]

リンカー導入法：ペプチドのＮ末端に導入できる。保護ペプチドに導入後脱保護する。
[Lp-H]

Linker introduction method: can be introduced at the N-terminus of the peptide. Deprotection after introduction into the protected peptide.
[Lp-H]

リンカー導入法：ペプチドのＮ末端に導入できる。ヒドロキシルアミノ基保護リンカーを保護ペプチドに導入後脱保護する。
[Lp-I]

Linker introduction method: can be introduced at the N-terminus of the peptide. A hydroxylamino group-protected linker is introduced into the protected peptide and then deprotected.
[Lp-I]

ペプチドのＣ末端に、例えば、以下の反応により導入できる。

It can introduce | transduce into the C terminal of a peptide by the following reaction, for example.

The combinations of spacers and respective linker parts, and the linker structures generated thereby are shown in Table 1 below.

  The method for synthesizing the polymer used in the present invention is not particularly limited. In the synthesis of the polymer, a spacer may be introduced after the polymer is synthesized, or a polymer may be synthesized by polymerizing a monomer having a spacer. Moreover, the polymer comprised from the unit which a peptide couple | bonds, and the unit which a peptide does not couple | bond together, the method of copolymerizing each monomer and then couple | bonding a peptide via the linker to the unit which couple | bonds a peptide is preferable. When the monomer terminal of the unit to which the peptide is bonded is a primary amine, the monomer in which the primary amino group is previously protected with a protecting group is radical-copolymerized with a monomer having a spacer that is a unit to which the peptide does not bind, and then the protecting group is removed. Is preferred. Then, the antimicrobial polypeptide containing polymer of this invention can be manufactured by couple | bonding a spacer and a peptide through a linker.

  Specific examples of monomers that do not bind peptides include poly (ethylene glycol) methyl ether (meth) acrylates, poly (ethylene glycol) (meth) acrylates, (meth) acrylic acid, n-butyl (meth) ) Acrylate, iso-butyl (meth) acrylate, sec-butyl (meth) acrylate, t-butyl (meth) acrylate, n-neopentyl (meth) acrylate, iso-neopentyl (meth) acrylate, sec-neopentyl (meth) acrylate , Neopentyl (meth) acrylate, n-hexyl (meth) acrylate, iso-hexyl (meth) acrylate, heptyl (meth) acrylate, n-octyl (meth) acrylate, iso-octyl (meth) acrylate, 2-ethylhexyl (meth) Acry Rate, n-nonyl (meth) acrylate, iso-nonyl (meth) acrylate, n-decyl (meth) acrylate, iso-decyl (meth) acrylate, n-dodecyl (meth) acrylate, iso-dodecyl (meth) acrylate, n-tridecyl (meth) acrylate, iso-tridecyl (meth) acrylate, n-tetradecyl (meth) acrylate, iso-tetradecyl (meth) acrylate, n-pentadecyl (meth) acrylate, iso-pentadecyl (meth) acrylate, n- Hexadecyl (meth) acrylate, iso-hexadecyl (meth) acrylate, n-octadecyl (meth) acrylate, iso-octadecyl (meth) acrylate, cyclohexyl (meth) acrylate, isobornyl (meth) acrylate, (meth) acrylic N-methyl (meth) acrylamide, N-ethyl (meth) acrylamide, N-propyl (meth) acrylamide, N-isopropyl (meth) acrylamide, N, N-dimethyl (meth) acrylamide, N, N-diethyl ( (Meth) acrylamide, N-hydroxymethyl (meth) acrylamide, N- (2-hydroxyethyl) (meth) acrylamide, N, N-bis (hydroxymethyl) (meth) acrylamide, N, N-bis (2-hydroxyethyl) ) (Meth) acrylamide and the like.

The monomer in which the primary amino group is previously protected with a protecting group is not particularly limited in structure, but the following formula (46) (wherein R ₂ is a hydrogen atom or a methyl group, V is a spacer containing an alkylene glycol residue, T Represents a protective group), the (meth) acrylic group preferably has a structure via a spacer V containing an alkylene glycol residue.

  The protecting group T is not limited as long as it can protect the amino group, and any protecting group can be used. Among them, a t-butoxycarbonyl group (Boc group) or a benzyloxycarbonyl group (Z group, Cbz group), 9-fluorenylmethoxycarbonyl group (Fmoc group) and the like are preferable. Deprotection can be performed under general conditions using trifluoroacetic acid, hydrochloric acid, anhydrous hydrogen fluoride, or the like.

  Specific examples of the monomer include those represented by the following formula (47).

  As a polymer synthesis solvent used in the present invention, any solvent may be used as long as each monomer can be dissolved. For example, alcohols such as methanol, ethanol, isopropanol, n-butanol, t-butyl alcohol, and n-pentanol, Benzene, toluene, tetrahydrofuran, dioxane, dichloromethane, chloroform, cyclohexanone, N, N-dimethylformamide, dimethyl sulfoxide, methyl acetate, ethyl acetate, butyl acetate, methyl ethyl ketone, methyl butyl ketone, ethylene glycol monoethyl ether, ethylene glycol monomethyl ether, Examples thereof include ethylene glycol monobutyl ether. These solvents may be used alone or in combination of two or more.

As the polymerization initiator, a normal radical initiator can be used. For example, 2,2′-azobisisobutylnitrile (hereinafter referred to as “AIBN”), 1,1′-azobis (cyclohexane-1-carbonitrile), etc. Organic peroxides such as azo compounds, benzoyl peroxide and lauryl peroxide.
In addition, the weight average molecular weight of the polymer used in the present invention is preferably 3,000 or more. By setting the weight average molecular weight to 3,000 or more, separation from the monomer is facilitated, and the fixing force to the fiber is further increased. More preferably, it is 10,000 or more.

  A linker site (functional group) can be introduced into the polymer (including the spacer) for the coupling reaction with the peptide. Although there is no restriction | limiting in particular as a functional group, For example, a halogen, a carboxyl group, an aldehyde group, an isocyanate group, a thioester group, a bromoacetyl group, a maleimide group, a thiol group etc. can be mention | raise | lifted. The linker to be introduced can be appropriately selected according to the functional group at the terminal of the peptide to be bound. When the bond destination is an amino group, it is preferable to introduce a halogen, a carboxyl group, an aldehyde group or an isocyanate group. When the bond destination is a carboxyl group, an amino group and a hydroxyl group are preferable. When the bonding destination is a thiol group, for example, an SH group of cysteine, a halogen, a thioester group, an aldehyde group, a bromoacetyl group, or a maleimide group is preferable. In particular, when a cysteine residue is introduced at the peptide end as a linker site, it is preferable to introduce a maleimide group or a halogen group at the end of the spacer.

  The peptide binding reaction to the spacer is not particularly limited and can be carried out by condensation reaction, thioetherification reaction, esterification reaction, Huisgen reaction, Michael addition reaction, oximation reaction, etc., but preferably the C-terminal of the peptide Alternatively, a Cys group or thiol group is introduced at the N-terminus, and a maleimide group in the polymer is used, a maleimide-thiol Michael addition reaction, or a thioetherification reaction is used with a halogen group in the polymer. Can be done

  For example, when the peptide has a Cys group at the C-terminus or N-terminus, the peptide-containing polymer is preferably prepared in a solvent that dissolves the polymer having a maleimide unit and is miscible with water. For example, by dissolving a polymer in alcohols such as methanol, ethanol, isopropanol, tetrahydrofuran, dioxane, N, N-dimethylformamide, dimethyl sulfoxide, etc., and reacting the peptide by adding a peptide solution dissolved in, for example, water. It can be carried out.

The following description is based on an example in which a maleimide group and a thiol group (cysteine) are used as linker sites, but the present invention is not limited to this.
Introduction of a maleimide group into a polymer can be performed using a compound having a maleimide group and a functional group capable of reacting with an amino group in one molecule. For example, maleimidopropionic acid N-succinimidyl, N- (α-maleimidoacetoxy) succinimide ester, N- (4-maleimidobutyryloxy) succinimide, N- (4-maleimidobutyryloxy) sulfosuccinimide sodium salt, N- ( 6-maleimidocaproyloxy) succinimide, N- (8-maleimidocapryloxy) succinimide, N- (8-maleimidocapryloxy) sulfosuccinimide sodium salt, N- (11-maleimidoundecanoyloxy) succinimide, N- ( 11-maleimidoundecanoyloxy) sulfosuccinimide sodium salt and the like. These reagents may be used alone or in combination of two or more.

In one embodiment of the antibacterial peptide-containing polymer of the present invention, it is represented by the following formula (48) (wherein R ₁ is a hydrogen atom or a methyl group, n is a natural number, and V is a spacer). As described above, the peptide is linked to the maleimide unit of the monomer unit via a cysteine at the C-terminus or N-terminus. In the formula, S in -S-Cys represents S in a cysteine residue.

The antibacterial peptide-containing polymer of the present invention may be all the monomer unit, but the monomer unit and the following formula (49) (R ₁ is a hydrogen atom or a methyl group, n is a natural number, V May be a spacer).

  In the polymer of the present invention, the above formula (48) and the following formula (50) (n is a natural number and Y is a single bond or a linear or branched chain which may have a substituent having a molecular weight of 10 to 3000). Or an alkylene chain or a polyoxyalkylene chain, preferably a linear or branched alkylene chain or polyethylene glycol which may have a substituent, or a copolymer of the above formula (48) or the above formula ( 49) and the following formula (50) (n is a natural number).

  Preferable polymers include peptide-containing polymers represented by the following formula (51). Here, n1 is a natural number and m1 is 0 or a natural number.

  In the above example, the peptide can be bound to the polymer by adding a peptide solution in which the peptide is dissolved in a polymer solution in which the peptide is dissolved in a solvent using a Michael addition reaction. Moreover, the ratio of the peptide to the maleimide unit can be appropriately selected according to the purpose. For example, when the peptide is bound to all maleimide units, the coupling reaction is performed so that the molar ratio of the maleimide unit to the peptide is 1: 1. I do.

  In another aspect of the invention, an antimicrobial fiber is provided in which the peptide-containing polymer of the invention is applied onto the fiber. In the antibacterial fiber of the present invention, the polypeptide is immobilized on the fiber, the peptide is not easily detached in normal use, and the antibacterial property can be maintained even in repeated use.

In the present invention, the fibers are plant fibers such as cotton, hemp and silk, animal fibers such as wool, animal hair and silk, regenerated fibers such as rayon, semi-synthetic fibers such as acetate, and chemically synthetic fibers such as polyester, nylon and acrylic. , And complexes thereof, but are not particularly limited thereto.
Further, as the form of the fiber, a woven fabric or a knitted fabric made of staples, filament yarns and processed yarns thereof is used, but a fiber web may be used. The fiber web refers to non-woven fabric, tissue paper, wet tissue, paper such as food wrapping paper, and the like.

  The antibacterial fiber of the present invention can be used for various fiber products that require antibacterial properties, such as sanitary products. Since the antibacterial fiber of the present invention is also excellent in safety, it can be used for medical use and medical supplies such as medical bandages, sheets, gauze and clothes.

  In the method of the present invention, the peptide-containing polymer is fixed on the fiber, but the term “on the fiber” as used in the present invention means that a part of the peptide-containing polymer is a part of the fiber in addition to the case where the peptide-containing polymer is completely present on the surface of the fiber. It also includes the state of being in the middle. In the case where the target peptide activity, for example, the peptide is a modified peptide derived from beetle defensin, it is preferable that all or most of the peptide-containing polymer is present on the fiber surface in order to exhibit antibacterial activity.

In the present invention, the polymer coated on the cloth is preferably 10% by weight or less, more preferably 5% by weight or less, based on the weight of the cloth. In this case, the peptide, preferably the fabric 1 cm 0.001Myu mol 0.5μ mol per ^2, most preferably 1 cm 0.03 microns mol per ² from safety and economical point of view.

  The peptide-containing polymer of the present invention can be fixed to the fiber by coating the fiber surface with the peptide-containing polymer, for example, using a fiber processing technique such as water repellent processing or antistatic processing. it can. In other words, the immobilization of the polymerized peptide to the fiber can be performed using, for example, a method based on electrostatic interaction, a method of chemically bonding by a crosslinking reaction, or the like. The method of immobilization by electrostatic interaction is simpler. On the other hand, when chemically bonded by a cross-linking reaction, there is an advantage of stronger immobilization, which can be arbitrarily selected according to the purpose. It is.

In another aspect of the present invention, an emulsion containing the antimicrobial peptide-containing polymer of the present invention is provided.
In the present invention, for example, a hydrophobic polymer (an peptide-containing polymer) having an immobilized peptide can be emulsified alone or with a surfactant or the like to form an emulsion that is a fiber processing agent. The polymer emulsion can also be prepared by a conventionally known emulsion polymerization method. For example, a monomer can be dispersed in water with a surfactant, and emulsion polymerization can be carried out by adding a polymerization initiator, a chelating agent, an oxygen scavenger, a molecular weight modifier and the like. Conventionally known surfactants, polymerization initiators, chelating agents, oxygen scavengers, and molecular weight modifiers can be used.

  Thereby, it is also possible to sell or distribute in an emulsion state. Moreover, the fiber in which the peptide of the present invention is immobilized can be easily produced by applying the emulsion thus prepared to the fiber and drying it. In the case of a water-soluble polymer, it can be sold or distributed in a state dissolved in water, and can be easily mixed with a cross-linking agent such as blocked isocyanate during processing and fixed to a cloth. .

In still another aspect of the present invention, there is provided a method for producing an antibacterial fiber having an antibacterial peptide immobilized, comprising the following steps.
Step (1); the following formula (1b):

（式中Ｒ₁及びＲ₃は、それぞれ独立に水素原子又はメチル基であり、ｎは自然数であり、ｍ2は０又は自然数であり、Ｕ₂及びＵ₃は、それぞれ独立にＮＨ又はＯであり、Ｖは、単結合又はスペーサーであり、Ｗoは、ペプチド又は誘導体化したペプチドと反応する官能基であり、Ｙは、単結合又は分子量１０〜３０００の置換基を有してもよい直鎖又は分岐状のアルキレン鎖又はポリオキシアルキレン鎖である。）で表されるポリマーを含む溶液を繊維にコーティングする工程、及び
工程（２）；下記式：

(Wherein R ₁ and R ₃ are each independently a hydrogen atom or a methyl group, n is a natural number, m 2 is 0 or a natural number, and U ₂ and U ₃ are each independently NH or O. , V is a single bond or a spacer, Wo is a functional group that reacts with a peptide or a derivatized peptide, and Y is a single bond or a straight chain that may have a substituent having a molecular weight of 10 to 3000 A branched alkylene chain or a polyoxyalkylene chain), a step of coating a fiber with a solution containing a polymer represented by formula (2);

（ここで、Peptideは、下記式で表されるアミノ酸配列：
−ＮＨ−Ｘ１−Ｌｅｕ−Ｘ２−Ｌｅｕ−Ｘ３−X４−Ｘ５−Ｘ６−Ｘ７−ＣＯＮＨ₂、又はＨ₂Ｎ−Ｘ１−Ｌｅｕ−Ｘ２−Ｌｅｕ−Ｘ３−X４−Ｘ５−Ｘ６−Ｘ７−ＣＯ−（式中、Ｘ１はＡｒｇ又はＡｌａであり、Ｘ２はＴｙｒ、Ａｒｇ又はＬｅｕであり、Ｘ３はＡｒｇ又はＡｌａであり、X４はＩｌｅ又はＬｅｕ又はＶａｌであり、Ｘ５はＧｌｙ又はＡｒｇであり、Ｘ６は、Ａｒｇ又はＬｙｓであり、Ｘ７は、Ａｒｇ又はＬｙｓである。ここで、Ａｒｇ−ＣＯＮＨ₂又はＬｙｓ−ＣＯＮＨ₂は、Ａｒｇ又はＬｙｓのカルボキシル基がアミド化していることを示す。）より選ばれる抗菌活性を有するポリペプチドである。）より選ばれ、Ｚは、水素原子、水酸基、Ｃｙｓ又はＷoと反応しリンカーを形成する官能基（例えば、アジド基、スルフヒドリル基、ハロゲン基、アルキン基、カルボニル基、ヒドロキシルアミノ基）を有する分子量５０〜５００の分子である。）
で表され抗菌性ポリペプチド又はその誘導体を含む溶液中に、ポリマーを含む溶液をコーティングした繊維を浸漬する工程、及び
工程（３）；１〜１０時間放置することによりポリマーにポリペプチドを結合させた後、繊維を洗浄して未反応のペプチド又はペプチド誘導体を除去する工程、
からなる抗菌性繊維の製造方法。

(Here, Peptide is an amino acid sequence represented by the following formula:
-NH-X1-Leu-X2- Leu-X3-X4-X5-X6-X7-CONH 2, or _{H 2 N-X1-Leu-} X2-Leu-X3-X4-X5-X6-X7-CO- ( In the formula, X1 is Arg or Ala, X2 is Tyr, Arg or Leu, X3 is Arg or Ala, X4 is Ile or Leu or Val, X5 is Gly or Arg, and X6 is Arg or Lys and X7 is Arg or Lys, where Arg-CONH ₂ or Lys-CONH ₂ indicates that the carboxyl group of Arg or Lys is amidated.) A polypeptide having Z is a molecular weight having a functional group (for example, azide group, sulfhydryl group, halogen group, alkyne group, carbonyl group, hydroxylamino group) that reacts with a hydrogen atom, a hydroxyl group, Cys or Wo to form a linker. 50 to 500 molecules. )
A step of immersing a fiber coated with a solution containing a polymer in a solution containing an antibacterial polypeptide or a derivative thereof, and step (3); allowing the polypeptide to bind to the polymer by standing for 1 to 10 hours And then removing the unreacted peptide or peptide derivative by washing the fiber,
The manufacturing method of the antimicrobial fiber which consists of.

  The binding reaction can be performed, for example, by reacting for 1 to 10 hours, preferably 2 to 10 hours, more preferably 3 to 6 hours. The concentration of the peptide solution used for the binding reaction can be appropriately selected depending on the reaction conditions such as the type of linker and temperature, and is, for example, 0.1 to 100 mM, preferably 1 to 10 mM.

The polymer to be coated (applied) to the fiber used in the method of the present invention may be any of the polymers prior to binding the polypeptides described herein. For example, as a preferred polymer, the following formula (6a) (wherein R ₁ and R ₃ are a hydrogen atom or a methyl group, n is a natural number, m is 0 or a natural number, U ₁ and U ₃ are , O or NH, and Y is a single bond or a linear or branched alkylene chain or polyoxyalkylene chain which may have a substituent having a molecular weight of 10 to 3000). However, it is not limited to this.

  EXAMPLES Hereinafter, although an Example demonstrates this invention in detail, this invention is not limited to these Examples.

(Example 1)
Peptide Synthesis Peptide A (RLYLRIGRR), Peptide 1 (CRLYLRIGRR) and Peptide 2 (RLYLRIGRRC) were synthesized by solid phase synthesis according to a conventional method.

(Example 2)
Synthesis of Monomer 1 To a two-necked flask (100 ml) equipped with a dropping funnel (3- {2- [2- (3-aminopropyl) -ethoxy] -ethoxy} -propyl) carbamic acid-t-butyl ester (1060 mg, 3.31 mmol), triethylamine (401 mg, 3.97 mmol) and tetrahydrofuran (25 mL) were added, and the mixture was stirred under ice cooling. A mixed solution of methacrylic acid chloride (913 mg, 3.97 mmol) and tetrahydrofuran (25 mL) was added dropwise thereto, and the mixture was stirred for 3 hours under ice cooling. Then, it stirred at room temperature for 8 hours. After completion of the reaction, the precipitate was removed, and the target product was isolated by silica gel column chromatography using ethyl acetate as a developing solvent. Rf value = 0.39 (silica gel, ethyl acetate) Yield 59%.
¹ H-NMR; (CDCl ₃ , 400 MHz) δ: 1.4-1.5 (9H, br, CH ₃ ), 1.7-1.8 (2H, m, NHCH ₂ CH ₂ ), 1.8-1.9 (2H, m, NHCH ₂ CH ₂ ), 2.05 (3H, s, OC CH ₃ ), 3.20-3.25 (2H, q, J = 6.24, NH CH ₂ ), 3.42-3.47 (2H, q, J-5.56, NH CH ₂ ), 3.51- 3.54 (2H, t, J = 5.96, O CH ₂ ), 3.55-3.65 (12H, m, O CH ₂ ) 5.29 (1H, s, CH ₃ C CH ₂ ), 5.71 (1H, s, CH ₃ C CH ₂ )

(Example 3)
Synthesis of Polymer 1 Monomer 1 (900 mg, 2.32 mmol), AIBN (7.6 mg, 0.046 mmol), and tetrahydrofuran (1 ml) were added to a nitrogen-substituted Schlenk tube (30 mL), and the mixture was stirred at reflux. After completion of the reaction, low molecular weight substances were removed by gel permeation chromatography. The obtained polymer was dissolved in dichloromethane (10 ml), trifluoroacetic acid (5 ml) was added, and the mixture was stirred at room temperature for 8 hours. After completion of the reaction, the solvent was distilled off under reduced pressure. The polymer was dissolved again in dichloromethane (10 ml), N-succinimidyl 3-maleimidopropionate (122.4 mg, 0.46 mmol) and triethylamine (46.5 mg, 0.46 mmol) were added, and the mixture was stirred at room temperature for 3 hours. After completion of the reaction, the polymer 1 was purified by gel permeation chromatography to obtain the following polymer 1.
¹ H-NMR; (CDCl ₃ , 400 MHz) δ: 0.8-1.0 (3H, br, CH ₃ ), 1.4-1.9 (8H, br, CH ₂ ), 2.4-2.6 (2H, br, CO CH ₂ ), 3.1-3.4 (4H, br, NH CH ₂ ), 3.4-3.7 (12H, br, O CH ₂ ), 3.75-3.85 (2H, br, CON CH ₂ ) 6.7-6.8 (2H, br, CO CH )
(Soluble in water, DMF, chloroform, THF)

Example 4
Synthesis of Polymer 2 (Peptide-Containing Polymer) Polymer 1 (15.8 mg) synthesized in Example 3 was dissolved in DMF (250 μl). Peptide 1 (CRLYLRIGRR) (11.7 mg, 9.0 mmol) was dissolved in 1 ml of water, and this was mixed with a DMF solution of polymer 1 and reacted. Peptide unit: maleimide unit = 1: 4

(Example 5)
Polymer 3 (peptide-containing polymer) was synthesized in the same manner as polymer 2 except that peptide 2 (RLYLRIGRRC) was used as the peptide to be reacted. Peptide unit: maleimide unit = 1: 4

(Example 6)
Synthetic polymer 1 (3.5 mg, maleimide unit 8 μmol) of polymer 4 (peptide-containing polymer ) was dissolved in water (1.0 ml). Peptide 2 (RLYLRIGRRC) (21 mg, 16 μmol) was dissolved in 1 ml of water, this was mixed with an aqueous solution of polymer 1 and reacted. This was dialyzed overnight with a dialysis tube (molecular weight cut off 8000) to remove excess peptide.

(Example 7)
Fabrication of processed cloths 1 to 3 The polymer 2DMF-water mixed solution was coated on a cotton JIS-attached white cloth and dried under reduced pressure to prepare processed cloths 1 to 3.
The processed cloth 1 was adjusted so that the amount of peptide present was 0.3 μmol per cm ^{2 of} cloth.
The processed cloth 2 was produced in the same manner as the processed cloth 1 except that the amount of the peptide was adjusted to 0.1 μmol per 1 cm ^{2 of the} cloth.
The processed cloth 3 was produced in the same manner as the processed cloth 1 except that the amount of the peptide was adjusted to 0.03 μmol per 1 cm ^{2 of the} cloth.

(Example 8)
Fabrication of work cloths 4 to 6 Fabrication of work cloths 4 to 6 was performed in the same manner as work cloth 1 except that polymer 3 was used as the polymer to be coated. The abundance of the peptide was 0.3 μmol per 1 cm ² of the processed cloth 4, 0.1 μmol per 1 cm ^{2 of} the processed cloth 5, and 0.03 μmol of the processed cloth 6 per 1 cm ^{2 of the} cloth.

(Comparative Example 1)
Fabrication of work cloths 7 to 9 Fabrication of work cloths 7 to 9 was performed in the same manner as work cloth 1 except that polymer 1 was used as the polymer to be coated. Maleimide amounts present unit, the fabric 7 is 1.2μmol per fabric 1 cm ^2, the fabric 8 is 0.4μmol per fabric 1 cm ^2, the workpiece cloth 9 was 0.12μmol per fabric 1 cm ^2.

Example 9
Fabrication of work cloth 10 Fabrication of work cloth 10 was performed in the same manner as work cloth 1 except that polymer 4 was used as the polymer to be coated. The amount of peptide present was 0.3 μmol per cm ² .

(Example 10)
Antimicrobial activity test The antimicrobial activity test was done about the processed cloth 1-10 produced as mentioned above. A 1% volume of culture solution (1 cm x 1 cm) and a standard fabric, each containing a culture medium (Staphylococcus aureus NBRC 12732) containing Staphylococcus aureus NBRC 12732, cultured overnight in 0.8% NB medium. After culturing at 37 ° C. for 3 hours, the solution was put into 200 μl of a culture solution diluted to have an absorbance at a wavelength of 600 nm of 0.01), and cultured at 37 ° C. for 18 hours. It was shown as a logarithmic value obtained by dividing the number of viable bacteria in the standard cloth by the number of viable bacteria in the processed cloth after 18 hours of culture. The results are shown in Table 1. After the antibacterial test is performed, the cloth is vortex washed with 40 ml of ultrapure water (maximum speed shaking for 10 seconds x 3 times) and sterilized by autoclave (121 ° C, 20 minutes), and the same processed cloth is used for the antibacterial test. Went. Further, this washing-sterilization operation-antibacterial test was repeated. The results are shown in Table 2 below.

(Example 11)
Synthesis of Polymer 5 Monomer 1 (54 mg, 0.14 mmol), N-octadecylacrylamide (931 mg, 2.88 mmol), AIBN (7.6 mg, 0.046 mmol) and chloroform (1 ml) were added to a nitrogen-substituted Schlenk tube (30 mL) and refluxed. Stir. After completion of the reaction, low molecular weight substances were removed by gel permeation chromatography. The obtained polymer was dissolved in dichloromethane (10 ml), trifluoroacetic acid (5 ml) was added, and the mixture was stirred at room temperature for 8 hours. After completion of the reaction, the solvent was distilled off under reduced pressure. The polymer was dissolved again in dichloromethane (10 ml), N-succinimidyl 3-maleimidopropionate (38 mg, 0.14 mmol) and triethylamine (14.1 mg, 0.14 mmol) were added, and the mixture was stirred at room temperature for 3 hours. After completion of the reaction, it was purified by gel permeation chromatography. As a result of NMR, the substance ratio of maleimide unit: octadecyl unit was 1:14.

¹ H-NMR; (CDCl ₃ , 400 MHz) δ: 0.86-0.90 (42H, t, J = 7.04, CH ₃ ), 1.26 (420H, br, CH ₂ ), 1.47 (10H, br, CH ₂ ), 1.76 -2.70 (64H, br, CH ₂ ), 3.15 (30H, br, NH CH ₂ ), 3.42-3.82 (14H, br, O CH ₂ , NCH ₂ ), 6.73 (2H, s, COC H )

(Example 12)
Fabrication of processed fabrics 11 and 12 The polymer 5 produced in Example 11 was coated on a cotton JIS-attached white fabric so that the amount of maleimide units in the fabric was 0.03 μmol per cm ^{2 of} fabric, and dried under reduced pressure. This cloth was immersed in an aqueous solution (peptide concentration 3.6 mM) containing peptide 1 (CRLYLRIGRR) in the same mole as the maleimide unit, and left for 5 hours. Thereafter, the cloth was washed with ion-exchanged water to remove unreacted peptides, and a processed cloth 11 was produced.
A processed cloth 12 was produced in the same manner as the processed cloth 11 except that peptide 2 (RLYLRIGRRC) was used as a peptide to be bound to the maleimide unit.

(Comparative Example 2)
Fabrication of work cloth 13 Fabrication was performed in the same manner as the work cloth 11 except that an aqueous solution containing the same mole of cysteine as the maleimide unit was used as a bond to the maleimide unit.

(Example 13)
In the same manner as in Example 10, the antibacterial activity of the work cloths 11 to 13 was measured. The results are shown in Table 3 below.

As a result, activity was observed in the processed cloths 1 to 6 and 10 to which the peptide-containing polymer was applied and the peptide was bound, but the processed cloths 7 to 9 not bound to the peptide had no activity at all. . Comparing the processed fabrics 1 to 3 and 4 to 6, it was found that the N-terminal bond type has slightly stronger antibacterial activity.
In addition, the activity was recognized in the processed cloths 11 and 12 to which the peptide was reacted after the polymer was applied to bind the peptide, but the activity was not recognized in the processed cloth 13 to which the peptide was not bonded.

(Example 14)
Synthesis of peptide with Cys linker Peptide 3 terminated with a Cys linker (CRLLLRVGRR-NH ₂ ) was synthesized by a solid phase synthesis method according to a conventional method.

(Actual example 15)
Synthesis of monomer 2
3-Bromopropylamine hydrobromide (2.15 g, 9.8 mmol) was dissolved in 50 mL of pyridine and stirred under ice cooling. A solution of methacrylic acid chloride (1.54 g, 14.7 mmol) dissolved in THF (30 mL) was added dropwise thereto and stirred overnight. After completion of the reaction, pyridine was distilled off to such an extent that it did not dry and separated with water and chloroform. The chloroform phase was concentrated and purified by gel permeation chromatography to give N- (3-bromopropyl) -2-methacrylamide. Yield 30%
¹ H-NMR; (CDCl ₃ , 400 MHz) δ: 2.22 (3H, s, CH ₃ ), 2.22-2.30 (2H, quin, J = 7.7, CH ₂ ), 3.86-3.90 (2H, t, J = 8.0 , CH ₂ ), 4.66-4.69 (2H, t, J = 7.4, CH ₂ ), 5.91 (1H, s, CH ₃ C CH ₂ ), 6.55 (1H, s, CH ₃ C CH ₂ )

(Example 16)
Synthesis of monomer 3
3-Bromo-1-propanol (1.39 g, 10 mmol) was dissolved in 10 ml of dehydrated toluene and stirred under a nitrogen atmosphere. To this was added dropwise 2-isocyanatoethyl methacrylate (1.55 g, 10 mmol), and the mixture was stirred at 60 ° C. overnight. The obtained by-product was removed by filtration to obtain monomer 2.
Yield 95%
¹ H-NMR; (CDCl ₃ , 400 MHz) δ: 1.95 (3H, s, CH ₃ ), 2.17 (2H, br, CH ₂ ), 3.45-3.54 (4H, br, Br CH ₂ , NC H ₂ ), 4.20-4.25 (4H, br, O CH ₂ ), 5.60 (1H, s, CH ₃ C CH ₂ ), 6.13 (1H, s, CH ₃ C CH ₂ )

(Example 17)
Synthesis of polymer 6
N- (3-bromopropyl) -2-methacrylamide (monomer 2) (82 mg, 0.4 mmol) and poly (ethylene glycol) methyl ether methacrylate Mn300 (6.0 g, 20 mmol) were dissolved in 20 mL of ethanol, AIBN (67 mg, 0.4 mmol) was added. The inside of the container was purged with nitrogen, and then stirred under reflux for 6 hours. Dialysis was performed for 48 hours with a dialysis tube (molecular weight cut off 3500) to remove unreacted monomers, and polymer 6 was obtained. In the Beilstein reaction, a flame reaction derived from halogen was observed.

Synthetic monomer 3 of polymer 7 (305 mg, 1.04 mmol) and poly (ethylene glycol) methyl ether methacrylate Mn300 (15.6 g, 52 mmol) were dissolved in 50 mL of ethanol, and initiator AIBN (174 mg, 1.06 mmol) was added. The inside of the container was purged with nitrogen, and then stirred under reflux for 6 hours. Dialysis was performed for 48 hours with a dialysis tube (molecular weight cut off 3500) to remove unreacted monomers, and polymer 7 was obtained.

(Example 18)
Synthesis of polymer 8 (peptide-containing polymer) Polymer 6 (8928 mg, 0.587 mmol in terms of Br) and peptide 3 with Cys linker (CRLLLRVGRR) (1062 mg, 0.587 mmol) were dissolved in 58.7 ml of DMF, and tertiary butoxypotassium (395.2 mg, 3.522 mmol) was added. The mixture was heated and stirred at 40 ° C. for 4 hours and 30 minutes. It was confirmed by TLC that the starting peptide had disappeared. The solution was diluted with 500 ml of DCM, and the precipitated salt was removed by celite filtration. The filtrate was concentrated with an evaporator and vacuum-dried to obtain polymer 8, which is a polymer in which peptide 3 is bound to polymer 6.

Synthesis of polymer 9 (peptide-containing polymer) Polymer 7 (2.04 g, 0.13 mmol in terms of Br) and peptide 3 with a Cys linker (CRLLLRVGRR) (235.4 mg, 0.13 mmol) were dissolved in 10 ml of DMF, and tertiary butoxy potassium (87.5 mg, 0.78 mmol) was added. The mixture was heated and stirred at 40 ° C. for 7 hours. It was confirmed by TLC that the starting peptide had disappeared. The solution was diluted with 50 ml of DCM, and the precipitated salt was removed by filtration through celite. The filtrate was concentrated with an evaporator and vacuum-dried to obtain polymer 9, which is a polymer in which peptide 3 is bound to polymer 7.

(Example 19)
Preparation of Peptide-Containing Polymer Processing Agents A and B Water was added to the polymer 8 (4.8 g) prepared in Example 18 to make a total amount of 121 g. Stir overnight and completely disperse to prepare Processing Agent A.
Processing agent A was diluted 10 times with distilled water to obtain processing agent B.
In addition, water was added to polymer 7 (346 mg) to make the total amount 17.3 g, and the mixture was stirred overnight to be completely dispersed to obtain a processing agent C (a peptide-free polymer processing agent). This was used for reference experiments.

(Example 20)
Water was added to adjustment polymer 9 (2.08 g) of peptide-containing polymer processing agent D to make the total amount 500 g. Stir overnight and completely disperse to prepare Processing Agent D.

(Example 21)
Fabrication of processed fabrics 14 to 16 Add 1.5 to 2.0 times the mass of processing agent A so that it penetrates the entire fabric, and remove excess processing agent with mangles, and dry at room temperature. A work cloth 14 was produced. The amount of peptide present was 0.03 μmol per cm ² . The processing agent was 4% based on the dough. In the ninhydrin reaction, it was confirmed that the dough was colored purple.
The processing agent B of 1.5 to 2.0 times the mass was added to the entire fabric, and the excess processing agent was removed with a mangle and dried at room temperature to produce a processed fabric 15. The amount of peptide present was 0.006 μmol / cm ² . The processing agent was 0.4% based on the dough.
Add 1.5 to 2.0 times the mass of processing agent C (reference example) to the entire fabric, and remove excess processing agent with mangles, dry at room temperature, and work cloth 16 (Reference Example) was produced. The processing agent was 6.8% based on the dough.
Fabrication of processed cloth 17 Add 1.5 to 2.0 times the mass of processing agent D to the entire fabric, and remove excess processing agent with mangles, dry at room temperature, and then process fabric. 17 was produced. The amount of peptide present was 0.006 μmol / cm ² . The processing agent was 0.8% based on the dough.

(Example 22)
Antibacterial test The processed fabric 17 was tested in the same manner as in Example 10. The results are shown in Table 4 below.

（実施例２３）
抗菌活性試験
上記の如くして作製した加工布１４〜１６について、抗微生物活性試験を行った。JIS L 1902 の定量試験方法に準拠して行った。培養後の生菌数測定法には、混釈平板培養法（コロニー法）を用いた。また、耐久性評価のための繰り返し洗濯試験は、JIS L 0217の洗い方103に規定する方法（40℃で5分間洗い、30℃で2回すすぎ洗いを2分間行う。脱水後、直射日光の当たらない場所でスクリーン乾燥）で、10回、20回、50回の繰り返し試験を行った。洗濯時には、ポリオキシエチレンアルキルエーテルを洗剤として用いた。ただし、静菌活性値は、（18時間培養後の標準布の生菌数の常用対数値−試験菌接種直後の標準布の生菌数の常用対数値）−（18時間培養後の加工布の生菌数の常用対数値−試験菌接種直後の加工布の生菌数の常用対数値）として示した。結果を以下の表５に示す。

(Example 23)
Antibacterial activity test The antimicrobial activity test was done about the processed cloth 14-16 produced as mentioned above. The measurement was performed in accordance with the JIS L 1902 quantitative test method. The pour plate culture method (colony method) was used as a method for measuring the viable cell count after the culture. In addition, the repeated washing test for durability evaluation was conducted according to the method specified in Washing Method 103 of JIS L 0217 (washing at 40 ° C for 5 minutes and rinsing twice at 30 ° C for 2 minutes. After dehydration, direct sunlight The test was repeated 10 times, 20 times, and 50 times by screen drying in a non-contact location. At the time of washing, polyoxyethylene alkyl ether was used as a detergent. However, the bacteriostatic activity value is (the common logarithm of the viable count of the standard cloth after 18 hours of culture-the common logarithm of the viable count of the standard cloth immediately after inoculation of the test bacteria)-(the processed cloth after 18 hours of culture) The common logarithm of the number of viable bacteria--the common logarithm of the number of viable bacteria of the processed cloth immediately after inoculation of the test bacteria). The results are shown in Table 5 below.

(Example 24)
Safety test of antibacterial peptide-containing polymer Using the polymer 8 produced in Example 18, a safety test was conducted for each of the following items.
1. Acute oral toxicity test Polymer 8 was administered to 5 males and 5 females of the test animals, but survived during the test period of 2 weeks. Was not recognized.
2. Mutagenicity test Polymer 8 was used as a sample, and the mutagenicity test was performed in accordance with the mutagenicity test in the Industrial Safety and Health Act. As test strains, Salmonella typhimurium used TA1535, TA100, TA1537, TA98, and Escherichia coli used WP2uvrA. The result was negative. Further, the mutagenicity was negative in the reverse mutation test conducted for 5 strains at this concentration.
3. Skin irritation test A test was conducted using processing agent A containing polymer 8. It was carried out by a method in accordance with "Standard Practice ASTM F719-81 ^-1996 on the Temporary skin hypersensitivity testing biomaterials in rabbits." PII value was less than 2.00 (weak stimulation).
4). Skin sensitization test Using polymer 8, guinea pigs were used in accordance with "Test method for biological safety assessment of medical devices" (Ministry of Health, Labor and Welfare, Medical Number Examination No. 36, Medical Device Examination No. 36). This was performed by the maximization method. The sensitization rate was 0%.

  From the above results, it was shown that the antibacterial peptide-containing polymer and antibacterial fiber of the present invention are safe and can be used in medical applications.

  The above description is merely illustrative of the objects and objects of the present invention and is not intended to limit the scope of the appended claims. Various changes and substitutions to the described embodiments will be apparent to those skilled in the art from the teachings described herein, all without departing from the scope of the appended claims. Range.

Claims (22)

Following formula (1)

Wherein R ₁ is a hydrogen atom or a methyl group, U ₁ is a single bond, O or NH, V is a single bond or a spacer, W is a single bond or an amide group, a thioether group, A linker having a thioester group, an oxime group, a 1,2,3-triazole group, a carbonyl group or an ester group, and Peptide is an amino acid sequence represented by the following formula:
-NH-X1-Leu-X2- Leu-X3-X4-X5-X6-X7-CONH 2, or _{H 2 N-X1-Leu-} X2-Leu-X3-X4-X5-X6-X7-CO- ( In the formula, X1 is Arg or Ala, X2 is Tyr, Arg or Leu, X3 is Arg or Ala, X4 is Ile, Leu or Val, X5 is Gly or Arg, and X6 is Arg Or Lys and X7 is Arg or Lys, where Arg-CONH ₂ or Lys-CONH ₂ indicates that the carboxyl group of Arg or Lys is amidated.) It is a peptide. )
The antibacterial peptide containing polymer containing the repeating unit represented by these. Further, the following formulas (2) and (3)

(Wherein R ₂ and R ₃ are each independently a hydrogen atom or a methyl group, U ₂ is a single bond, O or NH, U ₃ is O or NH, and V is a single bond or A spacer, W ₁ is a hydrogen atom or a group consisting of all or part of the linker W to which a halogen may be added at the end, and Y has a single bond or a substituent having a molecular weight of 10 to 3000 It may be a linear or branched alkylene chain or polyoxyalkylene chain.)
The antibacterial peptide-containing polymer according to claim 1, comprising at least one repeating unit represented by: In the formulas (1) to (3), -W- represents the following formula:

(Wherein t is a natural number of 1 to 7), and Y may have a single bond or a substituent having a molecular weight of 10 to 3000, linear or branched The antibacterial peptide-containing polymer according to claim 1, wherein the polymer is a chain-like alkylene chain or polyethylene glycol chain. In the formula (1), -W-Peptide is represented by the following formula:

(Where Peptide is the same as above),

(Here, Peptide is the same as described above, and i is an integer of 1 to 15.),

(Here, Peptide is the same as described above, and i is an integer of 1 to 7.),

(Here, Peptide is the same as described above, i is an integer of 1 to 7, and j is an integer of 1 to 15.),

(Where Peptide is the same as above),

(Here, Peptide is the same as described above, and i is an integer of 1 to 15.),

(Here, Peptide is the same as above, i is an integer of 1 to 8, and j is an integer of 1 to 8.),

(Here, Peptide is the same as described above, i is an integer of 1 to 7, and j is an integer of 1 to 7.),

(Where Peptide is the same as above),

(Here, Peptide is the same as described above, and i is an integer of 2 to 7.),

(Wherein Peptide is the same as above, i is an integer of 1 to 7, and j is an integer of 2 to 7),

(Here, Peptide is the same as above, i is an integer of 1 to 15, and j is an integer of 1 to 10.),

(Where Peptide is the same as above, i is an integer of 1 to 8, and j is an integer of 1 to 7),

(Where Peptide is the same as above, i is an integer of 1 to 7, and j is an integer of 1 to 7),

(Here, Peptide is the same as described above, and i is an integer of 2 to 7.),

(Here, Peptide is the same as above, i is an integer of 1 to 10, and j is an integer of 1 to 15.),

(Where Peptide is the same as above and i is an integer of 2 to 7), and

(Here, Peptide is the same as described above, and i is an integer of 1 to 10.)
Selected from the group consisting of
In formula (2), -W ₁ is represented by formulas (31) to (38):

(Where i is an integer from 1 to 7, j is an integer from 1 to 8, k is an integer from 1 to 15, l is an integer from 0 to 8, and h is from 1 to 5) And X is Cl, Br or I)
The antibacterial peptide-containing polymer according to claim 2, which is selected from the group consisting of: In formula (1), -W-Peptide is represented by the following formula:

(Where Peptide is the same as above), and W _{1 in the} formula (2) is the following formula:

The antibacterial peptide-containing polymer according to claim 2.   The spacer V may have a linear or branched polyethylene glycol which may have a substituent having a weight average molecular weight of 50 to 1000, or a linear or branched which may have a substituent having a molecular weight of 10 to 500. The antibacterial peptide-containing polymer according to any one of claims 1 to 5, which is an alkylene chain or a combination thereof. Following formula (4)

And a repeating unit represented by the following formula (5):

The antibacterial peptide containing polymer containing the repeating unit represented by these. The following formulas (1), (2) and (3)

An antibacterial peptide-containing polymer comprising a repeating unit represented by
In the formula (1), U ₁ is O, -VW-Peptide is -PEG-S-Cys-Peptide (wherein PEG is polyethylene glycol having a weight average molecular weight of 10 to 2000) or _{-CH 2 CHOHCH 2 -S-Cys-} Peptide, or, in U ₁ is NH, is -V-W-Peptide, -Peptide, - (CH 2) 3 -S-Cys-Peptide, -CH 2 - (C _{2 H 4 O) 3 -C 3} H 6 -NH-COCH 2 -S-Cys-Peptide, and the following formula:

Selected from the group consisting of and
In the formula (3), U ₃ is O, -YH is-(CH ₂ ) ₁₁ -CH ₃ , or -PEG-R ₃ (wherein PEG is a polyethylene having a weight average molecular weight of 10 to 2000) a glycol, R ₃ is OH or CH _3.), or U ₃ is in NH, -Y is a single bond, an antimicrobial peptide-containing polymer according to claim 2.   The antibacterial peptide-containing polymer according to any one of claims 1 to 8, wherein the Peptide-containing unit is contained in the polymer at a ratio of 1 to 100 mol%.   The antibacterial peptide-containing polymer according to claim 9, wherein the Peptide-containing unit is contained in the polymer at a ratio of 0.4 to 20 mol%. The peptide is
-HN-Arg-Leu-Tyr- Leu-Arg-Ile-Gly-Arg-Arg-CONH 2, -NH-Arg-Leu-Arg-Leu-Arg-Ile-Gly-Arg-Arg-CONH 2, -NH -Ala-Leu-Tyr-Leu- Ala-Ile-Arg-Arg-Arg-CONH 2, -HN-Arg-Leu-Leu-Leu-Arg-Ile-Gly-Arg-Arg-CONH 2, -HN-Arg -Leu-Tyr-Leu-Arg- Val-Gly-Arg-Arg-CONH 2, -NH-Arg-Leu-Arg-Leu-Arg-Val-Gly-Arg-Arg-CONH 2, -NH-Ala-Leu -Tyr-Leu-Ala-Val- Arg-Arg-Arg-CONH 2, -HN-Arg-Leu Leu-Leu-Arg-Val- Gly-Arg-Arg-CONH 2, H 2 N-Arg-Leu-Tyr-Leu-Arg-Ile-Gly-Arg-Arg-CO-, H 2 N-Arg-Leu- Arg-Leu-Arg-Ile- Gly-Arg-Arg-CO-, H 2 N-Ala-Leu-Tyr-Leu-Ala-Ile-Arg-Arg-Arg-CO-, H 2 N-Arg-Leu- Leu-Leu-Arg-Ile- Gly-Arg-Arg-CO-, H 2 N-Arg-Leu-Tyr-Leu-Arg-Val-Gly-Arg-Arg-CO-, H 2 N-Arg-Leu- Arg-Leu-Arg-Val- Gly-Arg-Arg-CO-, H 2 N-Ala-Leu-Tyr-Leu-Ala-Val-Arg-Ar -Arg-CO-, and _{H 2 N-Arg-Leu-} Leu-Leu-Arg-Val-Gly-Arg-Arg-CO- according to any of claims 1 to 10 which is a peptide selected from the group consisting of Antibacterial peptide-containing polymer.   The antibacterial fiber by which the antibacterial peptide containing polymer in any one of Claims 1-11 was coated on the fiber. The antibacterial fiber according to claim 12, wherein the antibacterial peptide-containing polymer is present in an amount of 0.001 μmol or more and 0.3 μmol or less per 1 cm ² of the fabric in a peptide abundance.   Use of the antibacterial fiber according to claim 12 or 13 in medical use or sanitary use.   A method for immobilizing an antimicrobial peptide on a fiber by coating the antimicrobial peptide-containing polymer according to any one of claims 1 to 11 on the fiber.   The polymer emulsion containing the antibacterial peptide containing polymer in any one of Claims 1-11. Following formula (9)

Wherein R ₁ is a hydrogen atom or a methyl group, U ₁ is O or NH, V is a single bond or a spacer, Wo is a functional group that reacts with a peptide or a derivatized peptide. is there.)
In the polymer containing the repeating unit represented by the following formula (10):

(Here, Peptide is an amino acid sequence represented by the following formula:
-NH-X1-Leu-X2- Leu-X3-X4-X5-X6-X7-CONH 2, or _{H 2 N-X1-Leu-} X2-Leu-X3-X4-X5-X6-X7-CO- ( In the formula, X1 is Arg or Ala, X2 is Tyr, Arg or Leu, X3 is Arg or Ala, X4 is Ile or Leu or Val, X5 is Gly or Arg, and X6 is Arg or Lys and X7 is Arg or Lys, where Arg-CONH ₂ or Lys-CONH ₂ indicates that the carboxyl group of Arg or Lys is amidated.) Is a peptide having Z is a molecule having a molecular weight of 50 to 500 having a functional group that reacts with a hydrogen atom, a hydroxyl group, Cys or Wo to form a linker. )
A method for producing an antibacterial peptide-containing polymer by reacting the peptide represented by The following formulas (9) and (3)

Wherein R ₁ and R ₃ are each independently a hydrogen atom or a methyl group, U ₁ and U ₃ are each independently O or NH, V is a single bond or a spacer, Wo is A functional group that reacts with a peptide or a derivatized peptide, and Y is a single bond or a linear or branched alkylene chain or polyoxyalkylene chain that may have a substituent having a molecular weight of 10 to 3000.
In the polymer containing the repeating unit represented by the following formula (10):

(Here, Peptide is an amino acid sequence represented by the following formula:
-NH-X1-Leu-X2- Leu-X3-X4-X5-X6-X7-CONH 2, or _{H 2 N-X1-Leu-} X2-Leu-X3-X4-X5-X6-X7-CO- ( In the formula, X1 is Arg or Ala, X2 is Tyr, Arg or Leu, X3 is Arg or Ala, X4 is Ile or Leu or Val, X5 is Gly or Arg, and X6 is Arg or Lys and X7 is Arg or Lys, where Arg-CONH ₂ or Lys-CONH ₂ indicates that the carboxyl group of Arg or Lys is amidated.) Is a peptide having Z is a molecule having a molecular weight of 50 to 500 having a functional group that reacts with a hydrogen atom, a hydroxyl group, Cys or Wo to form a linker. )
A method for producing an antibacterial peptide-containing polymer by reacting the peptide represented by The repeating unit represented by the formula (9) is represented by the following formula (6).

(Here, R ₂ is a hydrogen atom or a methyl group.)
The method according to claim 17 or 18, wherein the peptide is Z in the formula (10) is Cys. The following steps:
Process (1): following formula (9)

(Wherein R ₁ is a hydrogen atom or a methyl group, U ₁ is O or NH, V is a single bond or a spacer, Wo is a functional group that reacts with a peptide or a derivatized peptide) is there.)
A step of coating a fiber with a solution containing a polymer containing a repeating unit represented by formula (2): and the following formula:

(Here, Peptide is an amino acid sequence represented by the following formula:
-NH-X1-Leu-X2- Leu-X3-X4-X5-X6-X7-CONH 2, or _{H 2 N-X1-Leu-} X2-Leu-X3-X4-X5-X6-X7-CO- ( In the formula, X1 is Arg or Ala, X2 is Tyr, Arg or Leu, X3 is Arg or Ala, X4 is Ile or Leu or Val, X5 is Gly or Arg, and X6 is Arg or Lys and X7 is Arg or Lys, where Arg-CONH ₂ or Lys-CONH ₂ indicates that the carboxyl group of Arg or Lys is amidated.) Is a peptide having Z is a molecule having a molecular weight of 50 to 500 having a functional group that reacts with a hydrogen atom, a hydroxyl group, Cys or Wo to form a linker. )
A step of immersing the fiber coated with the polymer solution in a solution containing the antibacterial peptide or derivative thereof represented by: and step (3): after allowing the peptide to bind to the polymer by leaving it for 1 to 10 hours, Washing the fibers to remove unreacted peptides or derivatives thereof;
The manufacturing method of the antimicrobial fiber which consists of. The following steps:
Step (1): Formulas (9) and (3) below

Wherein R ₁ and R ₃ are each independently a hydrogen atom or a methyl group, U ₁ and U ₃ are each independently O or NH, V is a single bond or a spacer, Wo is A functional group that reacts with a peptide or a derivatized peptide, and Y is a single bond or a linear or branched alkylene chain or polyoxyalkylene chain that may have a substituent having a molecular weight of 10 to 3000.
A step of coating a fiber with a solution containing a polymer containing a repeating unit represented by formula (2): and the following formula:

(Here, Peptide is an amino acid sequence represented by the following formula:
-NH-X1-Leu-X2- Leu-X3-X4-X5-X6-X7-CONH 2, or _{H 2 N-X1-Leu-} X2-Leu-X3-X4-X5-X6-X7-CO- ( In the formula, X1 is Arg or Ala, X2 is Tyr, Arg or Leu, X3 is Arg or Ala, X4 is Ile or Leu or Val, X5 is Gly or Arg, and X6 is Arg or Lys and X7 is Arg or Lys, where Arg-CONH ₂ or Lys-CONH ₂ indicates that the carboxyl group of Arg or Lys is amidated.) Is a peptide having Z is a molecule having a molecular weight of 50 to 500 having a functional group that reacts with a hydrogen atom, a hydroxyl group, Cys or Wo to form a linker. )
A step of immersing the fiber coated with the polymer solution in a solution containing the antibacterial peptide or derivative thereof represented by: and step (3): after allowing the peptide to bind to the polymer by leaving it for 1 to 10 hours, Washing the fibers to remove unreacted peptides or derivatives thereof;
The manufacturing method of the antimicrobial fiber which consists of. The repeating unit represented by the formula (9) is represented by the following formula (6).

(Here, R ₂ is a hydrogen atom or a methyl group.)
The method for producing an antimicrobial fiber according to claim 20 or 21, wherein the peptide is Z in the formula 10 and Cys.