JP2009132654A - Synthetic polypeptide-containing aqueous cosmetic having reduced content of antibacterial component - Google Patents

Abstract

<P>PROBLEM TO BE SOLVED: To provide an aqueous cosmetic which excels in antibacterial properties even without substantially containing another antibacterial component and is stable over a long period. <P>SOLUTION: This aqueous cosmetic substantially does not contain an antibacterial component and contains at least a synthetic polypeptide which has an amino acid sequence represented by the formula: Pro-Y-Gly (wherein Y is Pro or Hyp) and forms a collagenous structure. The content of the antibacterial component, for example, may be not more than 100 ppm. The aqueous cosmetic may be in the form of an aqueous solution or an aqueous dispersion obtained by dissolving or dispersing the above synthetic polypeptide in water as a base. The concentration of the synthetic polypeptide may also be about 0.01 to 5 wt.%. The synthetic polypeptide may have a molecular weight peak in the range of not less than 10,000 in the molecular weight distribution. <P>COPYRIGHT: (C)2009,JPO&INPIT

Description

  Although the present invention uses water as a base and contains a collagen-like component, it has a high antibacterial property even when it does not substantially contain an antibacterial component, and has no risk of pathogen infection and is high. The present invention relates to a water-based cosmetic product having safety.

  Cosmetics contain organic components such as proteins and polysaccharides, and not only in aqueous solution or gel form, but also in creamy cosmetics, they contain a lot of water and are taken regularly from the container. Because it is repeatedly used, microorganisms such as bacteria are easy to propagate. For example, in recent years, many cosmetics contain naturally-derived components such as collagen. However, since collagen is a protein derived from mammals and fish, contamination with microorganisms associated with use can be achieved even after purification. Therefore, it is easy for bacteria to propagate. In addition, when microorganisms propagate in cosmetics, not only the cosmetics are contaminated, but the cosmetics are altered or physically deteriorated, or they are spoiled. In addition, the use of contaminated cosmetics may contaminate the skin or cause skin damage.

  Therefore, in general, antibacterial agents such as antibacterial agents or preservatives (for example, parabens, paraoxybenzoate esters, etc.), bactericides (for example, phenoxyethanol, etc.) are added to cosmetics to prevent the growth of germs. This ensures the long-term stability of cosmetics. In addition, in order to impart antibacterial or bactericidal action against a plurality of types of microorganisms to cosmetics, a plurality of types of antibacterial components may be used in combination or the amount of these antibacterial components used may be increased. However, these antibacterial ingredients may cause irritation or skin damage such as cosmetic irritation depending on the user's skin quality, amount of use, and combination of antibacterial ingredients used together. The safety of is questioned. For example, users with weak skin who have symptoms such as atopic dermatitis may react with antibacterial components such as preservatives to cause itching and the like because the barrier function inherent in the skin is weak. In view of such a background, in recent years, cosmetics which do not contain an antibacterial component called an antibacterial agent or an antiseptic or have a smaller blending amount are desired.

  In general, in cosmetics that do not contain antibacterial or preservatives as described above, the use period is shortened, so that the container size is reduced and the expiration date is recommended, storage in a refrigerator is recommended, When picking up the hand, the container is made a dropper type so that the container does not touch the hand directly. However, with these measures, it is difficult to prevent microbial contamination and propagation depending on the usage conditions of the user, so alcohols and organic acid salts with antibacterial or antiseptic effects other than conventional antibacterial or preservatives are added. By doing so, the propagation of microorganisms is often suppressed. For example, in Japanese Patent Application Laid-Open No. 2003-2810 (Patent Document 1), even if a cosmetic containing paraben, organic acid salt, ε-polylysine and / or a salt thereof is used with a small amount of paraben, It is disclosed that deterioration hardly occurs. Japanese Unexamined Patent Publication No. 2000-264830 (Patent Document 2) discloses a cosmetic composition containing a saccharide and a polyhydric alcohol and having usability, safety and antiseptic properties. However, alcohol and organic acid salts have relatively high safety (low toxicity) compared to conventional antibacterial or antiseptics such as parabens, but in order to impart sufficient antibacterial or antiseptic properties, It is necessary to use a large amount. In addition, when used in a large amount, ingredients in cosmetics (for example, proteins such as collagen) may be denatured, and depending on the user, the skin may be irritated or may cause skin problems. Therefore, even if such a low-toxic antibacterial component is used, it is difficult to achieve both antibacterial or antiseptic (bactericidal) or cosmetic stability and low skin irritation.

  On the other hand, in Japanese Patent Application Laid-Open No. 2003-111040 (Patent Document 3), an antibacterial cosmetic composition containing an iturin-based peptide as an active ingredient increases the stability of cosmetics against microbial contamination, and functions such as fat-soluble vitamins. Increasing the skin permeability of the ingredients is disclosed. However, in such cosmetics, properties required for cosmetics, for example, properties such as moisturizing properties, must be added with an active ingredient such as a moisturizing agent.

JP-A-2005-60314 (Patent Document 4) has at least an amino acid sequence represented by the formula Pro-Y-Gly (wherein Y represents Pro or Hyp), and has a collagen-like structure. Cosmetics composed of a synthetic polypeptide that forms a polymer is disclosed, and Japanese Patent Application Laid-Open No. 2007-137875 (Patent Document 5) discloses a porous or non-porous water-soluble cosmetic composed of the above synthetic polypeptide. It is disclosed. However, Patent Documents 4 and 5 do not disclose any antibacterial or antiseptic properties of aqueous cosmetics.
JP2003-2810A (Claim 1, paragraph number [0095]) JP 2000-264830 A (Claim 1, paragraph number [0036]) JP2003-1113040 (Claim 1, paragraph number [0124]) JP 2005-60314 A (Claim 1) JP 2007-137875 A (Claim 1)

  Accordingly, an object of the present invention is to provide an aqueous cosmetic product having excellent stability in which contamination by microorganisms is prevented for a long period of time without containing an antibacterial component that may cause skin damage. It is in.

  Another object of the present invention is to provide an aqueous cosmetic product having excellent safety and having no danger of causing infection by pathogens or transmission of pathogenic factors while having excellent properties of collagen.

  As a result of intensive studies to achieve the above-mentioned problems, the present inventors have found that an aqueous cosmetic containing a collagen-like synthetic polypeptide having a specific amino acid sequence has a high antibacterial or It was found that it has antiseptic properties and completed the present invention.

  That is, the aqueous cosmetic of the present invention is a cosmetic containing a polypeptide, substantially free of an antibacterial component, and at least the formula Pro-Y-Gly (wherein Y represents Pro or Hyp) as the polypeptide. And a synthetic polypeptide that forms a collagen-like structure. In the aqueous cosmetic, the proportion of the antibacterial component may be 100 ppm or less (for example, 10 ppm or less). The aqueous cosmetic may have a form of an aqueous solution or aqueous dispersion in which the synthetic polypeptide is dissolved or dispersed in water as a base, and the concentration of the synthetic polypeptide is about 0.01 to 5% by weight. May be. The aqueous cosmetic is usually sterilized. The synthetic polypeptide may have a molecular weight peak in a range of 10,000 or more in the molecular weight distribution. The synthetic polypeptide includes a polypeptide (I) composed of units represented by the following formulas (1) to (3), a peptide unit having an amino acid sequence represented by the following formula (4), and the following formula: It may be at least one polypeptide selected from the polypeptide (II) including the peptide unit having the amino acid sequence represented by (5).

[-(OC- (CH ₂ ) _m -CO) _p- (Pro-Y-Gly) _n- ] _a (1)
[-(OC- (CH ₂ ) _m -CO) _q- (Z) _r- ] _b (2)
[-HN-R-NH-] _c (3)
(In the formula, m is an integer of 1 to 18, p and q are the same or different and 0 or 1, Y represents Pro or Hyp, and n represents an integer of 1 to 20. Z represents an amino acid residue or 2 to 2. Represents a peptide chain consisting of 10 amino acid residues, r represents an integer of 1 to 20, R represents a linear or branched alkylene group, and the ratio (molar ratio) between a and b is a / B = 100/0 to 30/70, c = 1 when p = 1 and q = 0, c = b when p = 0 and q = 1, p = 1 and q = 1 C = a + b when c = 0 and c = 0 when p = 0 and q = 0.)
-Pro-Y-Gly- (4)
(Where Y is the same as above)
-Pro-V-Gly-W-Ala-Gly- (5)
(In the formula, V represents Gln, Asn, Leu, Ile, Val or Ala, and W represents Ile or Leu.)
The synthetic polypeptide is composed of repeating units of peptide units represented by the following formula (1a). The proportion of the units is 80% by weight or more in the synthetic polypeptide, and the molecular weight distribution is 2 × 10 ^4. The polypeptide may have a molecular weight peak in a range of ˜500 × 10 ⁴ and at least a part of the synthetic polypeptide can form a triple helical structure.

-(Pro-Y-Gly) _n- (1a)
(In the formula, Y represents Pro or Hyp, and n represents an integer of 1 to 15)
In the present invention, a synthetic polypeptide having an amino acid sequence represented by at least the formula Pro-Y-Gly (wherein Y represents Pro or Hyp) and forming a collagen-like structure is added to the aqueous cosmetic. Thus, a method for preventing the growth of microorganisms in aqueous cosmetics (preventing or inhibiting the growth of microorganisms) is also included.

  In the present specification, the term “antibacterial component” means a component having antibacterial, bactericidal, antifungal and / or antiseptic action, and has antibacterial activity such as conventional antibacterial agents, bactericides and antiseptics. In addition to the active ingredient, it is used to include an environment-inhibiting ingredient that does not exhibit antibacterial activity and forms a growth-inhibiting environment for microorganisms. In addition, in this specification, “antibacterial” means an action or effect that inhibits or inhibits the growth of microorganisms (bacteria, fungi, yeast, etc.) or kills microorganisms, and suppresses or prevents the growth of microorganisms. It is used in the meaning including the function or effect to perform.

  The aqueous cosmetic of the present invention contains a synthetic polypeptide having a specific amino acid sequence and capable of forming a collagen-like structure, so that the content of antibacterial components that may cause skin damage is low or the antibacterial components Even if it does not contain substantially, the contamination by microorganisms is prevented for a long period of time, and it is excellent in stability. In addition, the water-based cosmetic has excellent properties of collagen, but has no risk of causing pathogen infection or transmission of pathogenic factors, and is excellent in safety.

  The aqueous cosmetic of the present invention is a cosmetic containing a polypeptide, substantially free of antibacterial components, and at least the formula Pro-Y-Gly (wherein Y represents Pro or Hyp) as the polypeptide. It contains a synthetic polypeptide having the amino acid sequence represented and forming a collagen-like structure.

(Synthetic polypeptide)
In the present specification, various amino acid residues are described by the following abbreviations.

Ala: L-alanine residue
Arg: L-arginine residue
Asn: L-asparagine residue
Asp: L-aspartic acid residue
Cys: L-cysteine residue
Gln: L-glutamine residue
Glu: L-glutamic acid residue
Gly: Glycine residue
His: L-histidine residue
Hyp: L-hydroxyproline residue
Ile: L-isoleucine residue
Leu: L-leucine residue
Lys: L-lysine residue
Met: L-methionine residue
Phe: L-phenylalanine residue
Pro: L-proline residue
Sar: Sarcosine residue
Ser: L-serine residue
Thr: L-threonine residue
Trp: L-tryptophan residue
Tyr: L-tyrosine residue
Val: L-valine residue In the present specification, according to a conventional method, the amino acid sequence of the peptide chain is determined by positioning the N-terminal amino acid residue on the left side and the C-terminal amino acid residue on the right side. Describe.

  The synthetic polypeptide has at least an amino acid sequence represented by the formula Pro-Y-Gly (wherein Y represents Pro or Hyp). Since this amino acid sequence contributes to the stability of the triple helical structure, various polypeptides can be used as the polypeptide as long as it forms a collagenous tissue (collagenous tissue) or a collagen-like structure. Specifically, such a polypeptide includes a polypeptide (I) composed of peptide units represented by the following formulas (1) to (3) and an amino acid sequence represented by the following formula (4): And a polypeptide (II) including a peptide unit having a peptide unit having an amino acid sequence represented by the following formula (5). These polypeptides can be used alone or in combination of two or more.

[-(OC- (CH ₂ ) _m -CO) _p- (Pro-Y-Gly) _n- ] _a (1)
[-(OC- (CH ₂ ) _m -CO) _q- (Z) _r- ] _b (2)
[-HN-R-NH-] _c (3)
-Pro-Y-Gly- (4)
-Pro-V-Gly-W-Ala-Gly- (5)
(In the formula, m is an integer of 1 to 18, p and q are the same or different and 0 or 1, Y represents Pro or Hyp, and n represents an integer of 1 to 20. Z represents an amino acid residue or 2 to 2. Represents a peptide chain consisting of 10 amino acid residues, r represents an integer of 1 to 20, R represents a linear or branched alkylene group, and the ratio (molar ratio) between a and b is a / b = 100/0 to 30/70, c = a when p = 1 and q = 0, c = b when p = 0 and q = 1, p = 1 and q = 1 C = a + b when there is, c = 0 when p = 0 and q = 0;
V represents Gln, Asn, Leu, Ile, Val or Ala, and W represents Ile or Leu. )
In the polypeptide (I), the peptide unit (1) is an essential unit, and the peptide unit (1) needs to contain a repeating sequence of Pro-Y-Gly. In view of the stability of the triple helical structure and the ease of synthesis, the repeating number n is 1 to 20, preferably 1 to 15, more preferably 1 to 10 (for example, 1 to 5), particularly 1 It may be about ~ 3. The number of repeats n corresponds to the number of amino acid repeats of the starting peptide in the preparation of the synthetic polypeptide (I). From the viewpoint of preparation, it is industrially advantageous to use a peptide having n of 1 to 4 (particularly 1 to 3) as a raw material because the polypeptide can be synthesized by a liquid phase method.

  In the formula (1), Y may be either Pro or Hyp, but more preferably Hyp from the viewpoint of the stability of the triple helical structure. Hyp is usually a 4Hyp (eg, trans-4-hydroxy-L-proline) residue.

Further, m indicating the repetition number of the methylene chain (CH ₂₎ is usually 1 to 18, preferably 2 to 12, more preferably 2 to 10 (particularly 2-6) about. P is 0 or 1.

  In the peptide unit (2), Z represents an amino acid residue or a peptide chain having an arbitrary sequence composed of 2 to 10 amino acid residues. Z is not particularly limited, but from the viewpoint of the physical and biological properties of the polypeptide, the peptide chain Z is usually Gly, Sar, Ser, Glu, Asp, Lys, His, Ala, Val, Leu, Arg. , Pro, Tyr, Ile selected amino acid residues or peptide chains consisting of 2-10 amino acid residues, especially Gly, Sar, Ser, Glu, Asp, Lys, Arg, Pro, Val It often has an amino acid residue or a peptide chain consisting of 2 to 10 amino acid residues. Peptide chain Z is Gly, Sar, Ser, Glu, Asp, Lys, Arg-Gly-Asp, Tyr-Ile-Gly-Ser-Arg, Ile-Lys-Val-Ala-Val, Val-Pro-Gly-Val -Gly, Asp-Gly-Glu-Ala, Gly-Ile-Ala-Gly, His-Ala-Val, Glu-Arg-Leu-Glu, Lys-Asp-Pro-Lys-Arg-Leu, Arg-Ser-Arg It is preferable to include the sequence represented by -Lys.

  R indicating the number of repetitions of the peptide chain Z is usually about 1 to 20, preferably about 1 to 10, more preferably about 1 to 5, from the viewpoint of ease of synthesis and physical or biological properties of the polypeptide. is there.

In the formula (2), m, which represents the number of repeating methylene chains (CH ₂ ), is 1 to 18, preferably 2 to 12, more preferably 2 to 10 (particularly 2 to 6), as in the formula (1). ) Q is 0 or 1.

In the above formula, m, n, and r usually correspond to the number of repetitions in the starting amino acid or peptide. In the formulas (1) and (2), when at least one of p and q is 1 (that is, including a dicarboxylic acid unit), the polypeptide is a unit represented by the formula (3) ( Diamine unit). In this unit (3), examples of the linear or branched alkylene group represented by R include a C _1-18 alkylene group (or alkylidene group) such as methylene, ethylene, propylene, trimethylene, and tetramethylene. It can be illustrated. The alkylene group R may be a linear methylene chain (CH ₂ ) _s (wherein s represents an integer of 1 to 18). Preferred R is a C _2-12 alkylene group (more preferably a C _2-10 alkylene group, particularly a C _2-6 alkylene group).

  The ratio (a / b) between peptide unit (1) and peptide unit (2) is 100/0 to 30/70 (molar ratio), preferably 100/0 to 40/60 (molar ratio), more preferably It is about 100/0 to 50/50 (molar ratio).

  Furthermore, the ratio of the unit (3) can be selected according to the value of p in the formula (1) and the value of q in the formula (2). When p = 1 and q = 0, c = a Yes, when p = 0 and q = 1, c = b. Also, when p = 1 and q = 1, c = a + b, and when p = 0 and q = 0, c = 0.

That is, the polypeptide (I) includes (a) a polypeptide composed of a repeating unit of a peptide unit [-(Pro-Y-Gly) _n- ] in which p = 0 in the formula (1), b) A peptide unit [-(Pro-Y-Gly) _n- ] in which p = 0 in the formula (1) and a peptide unit [-(Z) _r- ] in which q = 0 in the formula (2) And (c) a peptide unit [-(OC- (CH ₂ ) _m -CO) wherein p = 1 in the formula (1)] -(Pro-Y-Gly) _n- ] and a unit [-HN-R-NH-] represented by the formula (3) at a ratio (molar ratio) of 1: 1. (D) a peptide unit [-(OC- (CH ₂ ) _m -CO)-(Pro-Y-Gly) _n- ] wherein p = 1 in the formula (1) and the formula (2) q = 1 and is a peptide unit _{[- (OC- (CH 2)} m -CO) - (Z) r -] and units represented by the formula (3) [-HN-R-NH-] and the a: b: include polypeptides comprised of repeating units in a proportion of a + b (molar ratio).

On the other hand, the polypeptide (II) contains a peptide unit (4) having an amino acid sequence represented by -Pro-Y-Gly- as an essential unit. This unit (4) may form a repeating structure represented by-(Pro-Y-Gly) _d -in the polypeptide from the viewpoint of the stability of the triple helical structure. The repeating number d of this arrangement is, for example, about 1 to 20, preferably about 2 to 15, and more preferably about 3 to 10. This repeat number d corresponds to the number of amino acid repeats of the starting peptide in the preparation of the synthetic polypeptide (II). From the viewpoint of preparation, it is industrially advantageous to use a peptide having d of 1 to 4 (particularly 1 to 3) as a raw material because the polypeptide can be synthesized by a liquid phase method. Y may be either Pro or Hyp. As described above, Y may be a Hyp [usually 4Hyp (eg, trans-4-hydroxy-L-proline) residue due to the stability of the triple helical structure. ] Is preferable.

  The polypeptide (II) in the present invention contains a peptide unit (5) having an amino acid sequence represented by -Pro-V-Gly-W-Ala-Gly-.

  V may be any of Gln, Asn, Leu, Ile, Val, or Ala, but Gln, Asn, Leu, Val, Ala, particularly Gln, Leu are more preferable. W may be either Ile or Leu, but Ile is more preferred.

  The combination of V and W is, for example, a type in which V is selected from Gln, Asn, Leu, Ile, Val and Ala (for example, Gln or Leu), and a peptide in which W is Ile, or V is Gln, Examples thereof include a peptide selected from Asn, Leu, Ile, Val, and Ala (for example, Gln or Leu), and W is Leu.

  A combination of Y, V and W is a peptide in which Y is Hyp, V is Gln, Asn, Leu, Ile, Val and Ala (for example, Gln or Leu), W is Ile or Leu, Examples include a peptide in which Y is Pro, V is Gln, Asn, Leu, Ile, Val and Ala (for example, Gln or Leu), and W is Ile or Leu.

Furthermore, the polypeptide (II) may contain other amino acid residues and peptide chains (units) as long as the physical and biological properties of the polypeptide are not inhibited. Examples of other amino acid residues or peptide chains include peptide chains represented _by- (Z) _r-in the peptide unit (2). Polypeptide (II) is selected from the viewpoint of physical and biological properties, for example, Gly, Sar, Ser, Glu, Asp, Lys, His, Ala, Val, Leu, Arg, Pro, Tyr, Ile. Amino acid residues or peptide chains composed of 2 to 10 amino acid residues, in particular, amino acid residues selected from Gly, Sar, Ser, Glu, Asp, Lys, Arg, Pro, Val, or 2 In many cases, it has a peptide chain composed of 10 amino acid residues. Specifically, for example, Gly, Sar, Ser, Glu, Asp, Lys, Arg-Gly-Asp, Tyr-Ile-Gly-Ser-Arg, Ile-Lys-Val-Ala-Val, Val-Pro-Gly -Val-Gly, Asp-Gly-Glu-Ala, Gly-Ile-Ala-Gly, His-Ala-Val, Glu-Arg-Leu-Glu, Lys-Asp-Pro-Lys-Arg-Leu, Arg-Ser It preferably contains an amino acid residue or peptide chain represented by -Arg-Lys.

  In the polypeptide (II), the ratio (molar ratio) between the peptide unit (4) and the peptide unit (5) is (4) / (5) = 99/1 to 30/70, preferably 98 / It is 2-40 / 60, More preferably, it is about 95 / 5-50 / 50.

  The ratio (molar ratio) between the total amount of the peptide unit (4) and the peptide unit (5) and the other peptide units is the former / the latter = 100 / 0-50 / 50, preferably 100 / 0-60. / 40, more preferably about 100/0 to 70/30.

Among the synthetic polypeptides, in particular, a polypeptide composed of a repeating unit of the peptide unit [-(Pro-Y-Gly) _n- ] (1a) wherein p = 0 in the formula (1), particularly a peptide unit A polypeptide composed of a repeating unit of [— (Pro-Hyp-Gly) _n −] (1b) is preferred. Further, a polypeptide in which the repeating unit n in the formulas (1a) and (1b) is an integer of 1 to 15, preferably an integer of 1 to 10 (for example, using a peptide having a repeating number n in such a range as a raw material. The resulting polypeptide is preferred. The ratio of the repeating unit is 50% by weight or more (for example, about 60 to 100% by weight) in the synthetic polypeptide, preferably 70% by weight or more (for example, about 70 to 95% by weight), and more preferably 80% by weight. % Or more (for example, about 80 to 90% by weight). Further, among these synthetic polypeptides, the molecular weight distribution has a molecular weight peak in the range of 5 × 10 ^{4 to} 100 × 10 ^{4 in} terms of globular protein converted by aqueous gel permeation chromatography (GPC), and is synthesized. A polypeptide in which at least a part of the polypeptide can form a triple helical structure is also preferred.

The synthetic polypeptide (polypeptide (I), (II), etc.) forms a chain polypeptide without forming a 6-membered ring by cyclization, and a solvent (water, sulfoxide such as dimethyl sulfoxide) , A hydrophilic solvent such as dimethylformamide, dimethylacetamide, N-methylpyrrolidone, or a mixed solvent thereof). In the molecular weight distribution, the polypeptide is, for example, 5000 or more (for example, about 5 × 10 ^{3 to} 1000 × 10 ⁴ ), preferably 10,000 or more (for example, in terms of globular protein conversion by aqueous gel permeation chromatography (GPC)). , 1 × ¹⁰ 4 ~500 × ¹⁰ about ^4), more preferably 20,000 or more (e.g., a peak molecular weight in the range of 2 × ¹⁰ 4 ~500 × ¹⁰ about ^4). In addition, the synthetic polypeptide has a molecular weight distribution of 50,000 or more (for example, about 5 × 10 ^{4 to} 500 × 10 ⁴ ), preferably 100,000 or more (for example, about 10 × 10 ^{4 to} 300 × 10 ⁴ ). More preferably, it may have a molecular weight peak in the range of 150,000 or more (for example, about 15 × 10 ^{4 to} 100 × 10 ⁴ ). The aqueous GPC measurement can be performed using an aqueous solution (for example, a phosphate buffer containing sodium chloride) as an eluent, and the standard substance is, for example, manufactured by Amersham Biosciences Co., Ltd. Gel Filtration LMW Calibration Kit (containing ribonuclease A, chymotrypsinogen A, ovalbumin, albumin, etc.), Gel Filtration HMW Calibration Kit (aldolase, catalase, ferritin, thyroglobulin, etc.) Containing) can be used.

  Furthermore, these polypeptides show a positive cotton effect at wavelengths of 220 to 230 nm and a negative cotton effect at wavelengths of 195 to 205 nm in the circular dichroism spectrum. Therefore, at least a part (that is, part or all) of the polypeptide can form a triple helical structure, forming a collagen-like polypeptide. The cotton effect refers to a phenomenon that occurs because the optical rotation material has different absorption coefficients for left and right circularly polarized light at a specific wavelength.

  These polypeptides can form a collagen tissue (collagenous tissue). Polypeptide chains forming the triple helical structure self-assemble to form fibrils of several nm to several tens of nm, and these fibrils are arranged to form a fiber structure of several μm to several tens of μm. can do. These can be observed with a transmission electron microscope, a scanning electron microscope, or an atomic force microscope.

  The polypeptide (I) and / or (II) may have biodegradability, particularly biodegradability. For example, the polypeptide (II) has collagenase degradability.

  Synthetic polypeptides may be physiologically or pharmacologically acceptable salts such as inorganic acids (hydrochloric acid, sulfuric acid, phosphoric acid, etc.), organic acids (acetic acid, trifluoroacetic acid, lactic acid, tartaric acid, maleic acid). Acids, fumaric acid, oxalic acid, malic acid, citric acid, oleic acid, palmitic acid, etc., metals (alkali metals such as sodium and potassium, alkaline earth metals such as calcium, aluminum, etc.), organic bases (trimethylamine, triethylamine, etc.) , T-butylamine, benzylamine, diethanolamine, dicyclohexylamine, arginine, etc.). These salt-forming compounds can be used alone or in combination of two or more. These salts can be obtained by ordinary salt formation reactions.

  Unlike the collagen derived from mammals, the synthetic polypeptide has no risk of infection or transmission of pathogens or pathogenic factors [for example, proteins converted to pathogenicity (for example, abnormal prions, etc.)]. Therefore, the polypeptide is highly safe and has high moisture retention and stability. In particular, the structure of the polypeptide (peptide bond, triple helix structure, etc.) can be maintained over a long period of time, and the structure of the polypeptide can be maintained even after sterilization (treatment by gamma irradiation, etc.). Excellent in properties.

(Method for producing polypeptide)
The synthetic polypeptide can be obtained by a conventional method in which amino acids and peptide segments constituting the polypeptide are subjected to a condensation reaction. In addition, the unit is not particularly limited as long as the unit is finally contained in the polypeptide.For example, the unit may be obtained by a method of condensing amino acids or a method of condensing amino acids with peptide segments. It is preferable to obtain a peptide component such as a peptide having an amino acid sequence or a derivative thereof and condensing the peptide component. The peptide chain of the peptide component can be synthesized by an ordinary peptide synthesis method, for example, a solid phase synthesis method or a liquid phase synthesis method. For peptide synthesis, conventional methods such as coupling methods using condensing agents, active ester methods (phenyl esters such as p-nitrophenyl ester (ONp) and pentafluorophenyl ester (Opfp), N-hydroxysuccinimide ester ( N-hydroxydicarboxylic acid imide ester such as ONSu), 1-hydroxybenzotriazole ester (Obt), etc.), mixed acid anhydride method, azide method and the like can be used. In a preferred method, at least a condensing agent (preferably a condensing agent described later, particularly a combination of a condensing agent and a condensing aid described later) is often used.

  Furthermore, in peptide synthesis, depending on the type of amino acid or peptide fragment, depending on the type of amino group, carboxyl group, or other functional group (guanidino group, imidazolyl group, mercapto group, hydroxyl group, ω-carboxyl group, etc.) Protection and elimination / removal of the protective group by catalytic reduction or strong acid treatment (anhydrous hydrogen fluoride, trifluoromethanesulfonic acid, trifluoroacetic acid, etc.) are repeated. For example, amino-protecting groups include benzyloxycarbonyl group (Z), p-methoxybenzyloxycarbonyl group (Z (OMe)), 9-fluorenylmethoxycarbonyl group (Fmoc), t-butoxycarbonyl group ( Boc), 3-nitro-2-pyridinesulfenyl group (Npys) and the like, and the protective group for the carboxyl group includes benzyloxy group (OBzl), phenacyloxy group (OPac), t-butoxy group (OBu ), Methoxy group (OMe), ethoxy group (OEt), and the like. An automatic synthesizer may be used for peptide synthesis. For details of polypeptide synthesis, reference can be made to JP-A-2005-60314.

  More specifically, the polypeptide (I) is prepared by, for example, condensing at least a peptide represented by the following formula (1a) or a derivative thereof (A).

X- (Pro-Y-Gly) _n -OH (1a)
(Wherein X represents H or HOOC— (CH ₂ ) _m —CO— (m is the same as above), and Y and n are the same as above).

  The peptide represented by the formula (1a) or its derivative (A) may be co-condensed with the peptide represented by the following formula (2a) or its derivative (B) to prepare a polypeptide.

X- (Z) _r -OH (2a)
(Wherein X represents H or HOOC— (CH ₂ ) _m —CO— (m is the same as above), and Z and r are the same as above).

Examples of the compound corresponding to X = HOOC— (CH ₂ ) _m —CO— include, for example, C such as malonic acid, succinic acid, glutaric acid, adipic acid, pimelic acid, suberic acid, azelaic acid, and sebacic acid. Examples thereof include _3-20 aliphatic dicarboxylic acids or acid anhydrides thereof. These compounds can be used alone or in combination of two or more. These compounds are also represented by the above (1a) and (2a) by reacting according to a conventional amide bond formation method (for example, a reaction using a tertiary amine or the like described later as a catalyst) reaction or the peptide synthesis method. A compound can be obtained.

  The ratio of the peptide or its derivative (A) to the peptide or its derivative (B) is, for example, the former (A) / the latter (B) = 100/0 to 30/70 (mol%), preferably 100/0. It is about -40/60 (mol%), More preferably, it is about 100 / 0-50 / 50 (mol%).

Furthermore, it is not necessary when X is H in the formula (1a) and / or formula (2a), but when X is HOOC— (CH ₂ ) _m —CO— (where m is the same as above). The peptide or its derivative (A) and / or the peptide or its derivative (B) is subjected to a cocondensation reaction with the compound (C) represented by the following formula (3a) in order to form an amide group. .

H ₂ NR-NH ₂ (3a)
(Wherein R is the same as above).

Examples of the compound represented by the formula (3a) include diamines corresponding to the formula (3), for example, C _1-18 alkylene diamine such as ethylene diamine, trimethylene diamine, propylene diamine, tetramethylene diamine, and hexamethylene diamine. And polyalkylene polyamines such as diethylenetriamine and hexamethylenetetramine. These compounds can be used alone or in combination of two or more.

The amount of the diamine compound (C) used is, for example, that one of the peptides or derivatives (A) and (B) thereof or a derivative thereof is X = HOOC— (CH ₂ ) _m —CO— (where m is In the case of having the same), the amount of the diamine compound (C) used is substantially 1 mol (for example, about 0.95 to 1.05 mol) with respect to 1 mol of the peptide or derivative thereof having such a group. ) It is necessary to use.

  In the preparation of polypeptide (II), a method of reacting a peptide component containing at least a peptide having the amino acid sequence includes (a) a peptide having both amino acid sequences represented by formulas (4) and (5) ( That is, a method of condensing a peptide component containing at least a peptide unit having a peptide unit having an amino acid sequence represented by formula (4) and a peptide unit having an amino acid sequence represented by formula (5)) And (b) a method of condensing a peptide component comprising at least a peptide having an amino acid sequence represented by formula (4) and a peptide having an amino acid sequence represented by formula (5).

  In the methods (a) and (b), peptides having both amino acid sequences represented by the formulas (1) and (2) can be used alone or in combination of two or more. In this method, the peptide component includes, in addition to the peptide, other peptides (for example, peptides having the amino acid sequence (1) and / or the amino acid sequence (2), the other amino acids, depending on the target polypeptide. A peptide containing a residue or a peptide chain) may be used.

  The condensation reaction of the peptide component is usually performed in a solvent. The solvent only needs to be able to dissolve or suspend the peptide component (partially or completely dissolve), and water and / or an organic solvent can be usually used. Examples of the solvent include water, amides (dimethylformamide, dimethylacetamide, hexamethylphosphoramide, etc.), sulfoxides (dimethylsulfoxide, etc.), nitrogen-containing cyclic compounds (N-methylpyrrolidone, pyridine, etc.), nitriles ( Examples include acetonitrile (such as acetonitrile), ethers (such as dioxane and tetrahydrofuran), alcohols (such as methyl alcohol, ethyl alcohol, and propyl alcohol), and mixed solvents thereof. Of these solvents, water, dimethylformamide, and dimethyl sulfoxide are frequently used.

  The reaction of these peptide components can usually be carried out in the presence of at least a dehydrating agent (dehydrating condensing agent) or a condensing agent. When the reaction is carried out in the presence of a dehydrating condensing agent and a condensing aid, dimerization or cyclization can be achieved. A polypeptide can be produced smoothly while suppressing the conversion.

  The dehydrating condensing agent is not particularly limited as long as dehydrating condensation can be efficiently performed in the solvent. For example, a carbodiimide condensing agent [diisopropylcarbodiimide (DIPC), 1-ethyl-3- (3-dimethylaminopropyl) -carbodiimide ( EDC = WSCI), 1-ethyl-3- (3-dimethylaminopropyl) -carbodiimide hydrochloride (WSCI · HCl), dicyclohexylcarbodiimide (DCC), etc.], fluorophosphate condensing agent [O- (7-azabenzotriazole -1-yl) -1,1,3,3-tetramethyluronium hexafluorophosphate, O-benzotriazol-1-yl-N, N, N ′, N′-tetramethyluronium hexafluorophosphate, benzo Triazol-1-yl-oxy-tris-pyrrolidinophosphonium hexafluorophosphate Benzotriazol-1-yl - tris (dimethylamino) phosphonium hexafluorophosphate product salt (BOP)], such as diphenylphosphoryl azide (DPPA) can be exemplified. These dehydration condensing agents can be used alone or in combination of two or more. A preferred dehydrating condensing agent is a carbodiimide condensing agent [for example, 1-ethyl-3- (3-dimethylaminopropyl) -carbodiimide, 1-ethyl-3- (3-dimethylaminopropyl) -carbodiimide hydrochloride].

  The condensation aid is not particularly limited as long as it promotes the reaction of the condensation agent. For example, N-hydroxypolycarboxylic imides [for example, N-hydroxysuccinimide (HONSu), N-hydroxy-5-norbornene N-hydroxydicarboxylic imides such as -2,3-dicarboxylic acid imide (HONB)], N-hydroxytriazoles [for example, N-hydroxybenzotriazoles such as 1-hydroxybenzotriazole (HOBt)], 3- Examples thereof include triazines such as hydroxy-4-oxo-3,4-dihydro-1,2,3-benzotriazine (HOObt), and 2-hydroxyimino-2-cyanoacetic acid ethyl ester. These condensation aids can also be used alone or in combination. Preferred condensation aids are N-hydroxydicarboxylic imides [such as HONSu], N-hydroxybenzotriazole or N-hydroxybenzotriazines [such as HOBt].

  The dehydration condensation agent and the condensation aid can be used in appropriate combination. Examples of the combination of the dehydrating condensing agent and the condensing aid include DCC-HONSu (HOBt or HOObt), WSCI-HONSu (HOBt or HOObt), and the like.

  The amount of the dehydrating condensing agent used is usually 0.7 to 5 mol, preferably 0.8 when a non-aqueous solvent not containing water is used per 1 mol of the total amount of the peptide component (including the diamine compound). It is about -2.5 mol, More preferably, it is about 0.9-2.3 mol (for example, 1-2 mol). In a solvent containing water (aqueous solvent), since the dehydration condensation agent is deactivated by water, the amount of the dehydration condensation agent used is usually 2 to 500 mol (for example, relative to 1 mol of the total amount of the peptide components). 2 to 50 mol), preferably 3 to 250 mol (for example, 5 to 25 mol), and more preferably about 4 to 125 mol (for example, 10 to 20 mol). The amount of the condensation aid used is, for example, 0.05 to 5 mol (for example, 0.5 to 5 mol), preferably 0.1, with respect to 1 mol of the total peptide component, regardless of the type of solvent. ~ 2 mol (e.g. 0.7-2 mol), more preferably 0.12-1.5 mol (e.g. 0.8-1.5 mol), especially 0.15-1 mol (e.g. , 0.17 to 0.5 mol).

  In the condensation reaction, the pH of the reaction system may be adjusted, or a base that does not participate in the reaction may be added. The pH can be adjusted usually using an inorganic base (sodium hydroxide, potassium hydroxide, sodium carbonate, sodium hydrogen carbonate, etc.), an organic base, an inorganic acid (hydrochloric acid, etc.) or an organic acid. The pH of the solution is adjusted to near neutral (pH = about 6 to 8). Examples of the base not involved in the reaction include tertiary amines such as trialkylamines such as trimethylamine, triethylamine and diisopropylethylamine, and heterocyclic tertiary amines such as N-methylmorpholine and pyridine. . The amount of such base used can usually be selected from a range of about 1 to 2 times the total number of moles of the peptide.

(Water-based cosmetics)
The form of the aqueous cosmetic product of the present invention is not particularly limited as long as it is a cosmetic product containing water as a main base (or carrier), and may be a gel aqueous cosmetic product, a milky aqueous cosmetic product, a creamy aqueous cosmetic product, or the like. However, it is usually in the form of an aqueous solution or aqueous dispersion in which a synthetic polypeptide is dissolved or dispersed in water as a base (liquid aqueous cosmetic (skin lotion)).

  The concentration of the synthetic polypeptide in the aqueous cosmetic is not particularly limited, and can be selected, for example, from the range of about 0.01 to 10% by weight, but is usually 0.01 to 5% by weight, preferably 0.05 to 3% by weight. %, More preferably about 0.1 to 2% by weight.

  The water-based cosmetic may not contain a base other than water, but may contain a base other than water as necessary depending on the form of the cosmetic. Examples of such bases include animal and vegetable oil bases (jojoba oil, olive oil, palm oil, camellia oil, macadamia nut oil, castor oil, squalane, etc.), mineral oil bases (liquid paraffin, polybutene, Silicone oil, etc.), synthetic oil bases (synthetic ester oils, synthetic polyether oils, etc.), water-soluble organic solvents [lower aliphatic alcohols (ethanol, isopropanol, etc.); alkylene glycols (ethylene glycol, polyethylene glycol, etc.); Liquid bases such as polyhydric alcohols such as glycerin and pentaerythritol; carboxylic acids such as lactic acid and sodium pyrrolidonecarboxylate]; fatty acid esters, higher alcohols, higher fatty acids, gel bases (polysaccharides such as gums and alginic acid) , Proteins such as royal jelly Quality, etc.) and the like solid or semi-solid bases such as. These bases may be used alone or in combination of two or more. The ratio of these bases is, for example, 0.01 to 20 parts by weight, preferably 0.05 to 10 parts by weight, and more preferably about 0.1 to 5 parts by weight with respect to 100 parts by weight of water. Also good.

  The aqueous cosmetic product of the present invention has high antibacterial properties. Therefore, the ratio of other antibacterial components is small, for example, 100 ppm or less (for example, about 0 to 100 ppm), preferably 50 ppm or less (for example, about 0 to 50 ppm), more preferably 10 ppm or less (for example, about 0 to 10 ppm). Especially, even if it is about 0-5 ppm (for example, 0.01-3 ppm), the antibacterial property of cosmetics can be maintained, and the stability of cosmetics can be maintained over a long period of time. That is, the cosmetic of the present invention contains a collagen-like polypeptide, which contains a component that is considered to be easy for microorganisms to propagate, and even though it is in a form in which aqueous microorganisms are extremely easy to propagate, a collagen-like polypeptide. Therefore, even if it does not contain other antibacterial components substantially, it is excellent in antibacterial properties. In addition, even when other antibacterial components are contained, the amount of use can be greatly reduced, so that irritation to the skin can be reduced while maintaining antibacterial properties, and the feeling of use is excellent.

  Examples of the antibacterial component as described above include phenolic compounds (such as phenol, chlorcresol, chlorxylenol, propylmethylphenol, resorcin, orthophenylphenol, paraoxybenzoate, thymol, cresol, parachlorophenol), organic acids [Organic acids, organic acid esters or salts thereof such as aromatic carboxylic acids {benzoic acid, sodium benzoate, paraoxybenzoic acid ester (paraben), salicylic acid, sodium salicylate, etc.}, aliphatic carboxylic acids {citrate, Aliphatic oxyacid salts such as malates; dehydroacetic acid, sodium dehydroacetate, sorbic acid, potassium sorbate, undecylenic acid monoethanolamide), halides [halocarbanilides (trichlorocarbanilides) , Trifluoromethyldichlorocarbanilide, etc.), chlorhexidines (chlorhexidine hydrochloride, chlorhexidine glucuronate, etc.), trichlorohydroxydiphenyl ether, triclosan, etc.), quaternary ammonium salts (eg benzalkonium chloride, distearylmethyl chloride) Ammonium, benzethonium chloride, alkyltrimethylammonium chloride, cetylpyridinium chloride, alkylisoquinolinium bromide, domifane bromide), amphoteric surfactants (alkyldiaminoethylglycine hydrochloride, laurylaminoethylglycine hydrochloride, lauroyl sarcosine sodium, etc.) , Cationic surfactants (benzalkonium chloride, benzethonium chloride, etc.), isothiazolinones (2-methyl-4-isothiazolin-3-one, 5- In addition to olo-2-methyl-4-isothiazolin-3-one, etc., isothiazolinone compounds described in JP-A-2006-273719), amine oxides (dimethyl lauryl amine oxide, dihydroxyethyl lauryl amine oxide, etc.), apo Lactoferrin, antibacterial or bactericidal basic protein or peptide (such as iturin peptide, surfactin peptide, protamine or salt thereof (such as protamine sulfate)), polylysine (ε-polylysine or salt thereof (inorganic or organic acid, etc.) ), Antibacterial metal compounds (such as metal compounds capable of producing silver ions, copper ions, etc.), antibacterial enzymes (proteases, lipases, oxidoreductases, carbohydrases, transferases, phytases, etc.), photosensitizers, and tyrams. Antibacterial active ingredients such as chloramine T, hinokitiol, jamal, glidant, dawicil; alcohols (alkanols optionally having a substituent such as ethanol, chlorobutanol, phenoxyethanol; ethylene glycol, ethylene glycol monoethyl ether, glycerin, di Ingredients that form a growth-inhibiting environment, such as glycerin, polyglycerin, aliphatic polyols such as glycerin monoethyl ether or ethers thereof) and sugars (such as sugar alcohols such as glucose, maltitol, and xylitol). Of these antibacterial components, paraoxybenzoic acid esters and phenoxyethanol are particularly widely used as preservatives for cosmetics.

  The aqueous cosmetic of the present invention only needs to contain the above-mentioned synthetic polypeptide and water as a base, and may not contain other components, but if necessary, conventional components used in cosmetic applications, for example, , Active ingredients and / or additives may be included.

  Examples of the active ingredient include humectants [sodium pyrrolidone carboxylate; amino acids (serine, glycine, threonine, alanine, etc.); polysaccharides (hyaluronic acid, sodium chondroitin sulfate, chondroitin heparin, etc.); proteins (vitronectin, fibronectin, keratin, Elastin, royal jelly, sericin, etc.], astringents (oxyacids such as citric acid, lactic acid, tartaric acid, or salts thereof; aluminum compounds such as aluminum chloride; zinc compounds such as zinc sulfate and sulfophenoxo zinc; proanthocyanidins Tannin-containing plant extracts such as hamamelis, birch, etc .; oyster extract, dio extract, horsetail extract, etc.), emollients (triglyceride oil, squalane, ester oil, etc.) Emulsions emulsified with nonionic emulsifiers such as cerides), emollients (salicylic acid or derivatives thereof, lactic acid, urea, etc.), antioxidants (tocopherols or derivatives thereof; polyphenols such as anthocyanins), UV absorbers, etc. And inorganic pigments that scatter UV rays, whitening agents (ascorbic acid or its derivatives, cysteine, placenta extract, arbutin, kojic acid, lucinol, ellagic acid, chamomile extract, etc.), antiperspirants (aluminum compounds, zinc compounds, tannins, etc.) Skin astringents), rough skin prevention agents (glycyrrhizinate, vitamins, etc.), anti-inflammatory agents (allantoin, guaiazulene, glycyrrhizic acid or salts thereof, glycyrrhetinic acid or salts thereof, ε-aminocaproic acid, tranexamic acid, ibuprofen, indomethacin , Oxide Lead or derivatives thereof; plant extracts such as Arnica extract), coenzymes (Coenzyme Q10, etc.), vitamins (vitamin A, vitamin B, vitamin C, vitamin D, vitamin E, vitamin K, etc.), amino acids (Tryptophan, cysteine, etc.), cell activators (riboflavin, pyridoxine, nicotinic acid, pantothenic acid, α-tocopherol, or derivatives thereof; plant extracts such as yukinoshita extract, etc.), active ingredients in freckles cosmetics (tyrosinase activity) Inhibitors, melanin reducing agents, etc.), active ingredients of acne cosmetics (such as sulfur, keratin softeners, anti-inflammatory agents, corticosteroids, sebum secretion inhibitors, etc.). The other active ingredients can be used alone or in combination of two or more. The proportion of the other active ingredient is 0.01 to 30 parts by weight, preferably 0.1 to 20 parts by weight, more preferably about 1 to 10 parts by weight, in terms of solid content, with respect to 100 parts by weight of the synthetic polypeptide. It may be.

  Examples of the additive include surfactants (anionic and nonionic surfactants), inorganic salts (sodium sulfate, sodium bicarbonate, potassium chloride, etc.), colorants, fibers (synthetic fibers such as nylon fibers, Natural fibers, etc.), abrasives (calcium hydrogen phosphate, calcium carbonate, anhydrous silicic acid, etc.), foaming agents (sodium lauryl sulfate, etc.), binders (the mucous substances exemplified above, for example, carboxymethyl cellulose, carboxymethyl cellulose sodium, carrageenan Etc.), opacifiers, fragrances (synthetic fragrances, essential oils, essential oil components, etc.), plant extracts, pH adjusters (bases such as sodium bicarbonate; acids such as sodium monohydrogen phosphate; borax, etc.), chelating agents Sequestering agent, solidifying agent, solubilizer, plasticizer, gelling agent (organic modified bentonite, etc.) Agents, organic solvents, reducing agents (thioglycolic acid or its salts, cysteine, etc.), a basic agent, oxidizing agents, cooling agents (such as menthol), and the like. The said additive can be used individually or in combination of 2 or more types. The ratio of the additive may be 0.001 to 10 parts by weight, preferably 0.01 to 5 parts by weight, or about 0.1 to 2 parts by weight with respect to 100 parts by weight of the synthetic polypeptide.

  In addition, you may use the said other base, an active ingredient, and an additive with the form of a salt. Such salts are preferably physiologically or pharmaceutically acceptable salts, such as organic acid salts (for example, carboxylates such as acetate, fumarate and citrate; organic sulfones such as methanesulfonate). Acid salts), inorganic acid salts (eg, hydrochloride salts), salts with organic bases (eg, salts with tertiary amines such as trimethylamine salts, ethanolamine salts, etc.), salts with inorganic bases (eg, Ammonium salts; alkali metal salts such as sodium salts; alkaline earth metal salts such as calcium salts; aluminum salts). Other bases, active ingredients, and additives may have an interaction property with the polypeptide as long as the properties of the polypeptide are not impaired. , Degradability, etc.).

(Method for producing water-based cosmetics)
The aqueous cosmetic product of the present invention can be produced by mixing at least the synthetic polypeptide, water as a base, and other components (other bases, active ingredients, additives, etc.) as necessary. Preferably, the aqueous cosmetic can be produced by dissolving or dispersing the synthetic polypeptide in water as a base. The concentration of the synthetic polypeptide in the aqueous cosmetic product may be adjusted by subjecting the reaction mixture obtained with the production of the synthetic polypeptide to ultrafiltration. For details of concentration adjustment by ultrafiltration, for example, JP-A-2007-137875 can be referred to.

  Aqueous cosmetics may be sterilized to maintain antibacterial properties. The sterilization treatment can be performed by a conventional sterilization treatment method (heat treatment (high-temperature sterilization treatment, low-temperature sterilization treatment, etc.), γ-ray irradiation, etc.). A preferred sterilization treatment method is a low temperature sterilization treatment or a γ-ray irradiation treatment. For example, in the pasteurization (or sterilization) treatment, the treatment is performed at a temperature of about 50 to 80 ° C. (preferably 55 to 70 ° C.) for 10 to 120 minutes, preferably 15 to 60 minutes, and more preferably about 20 to 40 minutes. May be. Synthetic polypeptides can be effectively sterilized because they do not deteriorate at relatively high temperatures.

  Moreover, in this invention, by adding at least the said synthetic polypeptide to aqueous cosmetics, the proliferation or growth of microorganisms in aqueous cosmetics can be prevented or inhibited, and such a method is also included in this invention. The form of the aqueous cosmetic product to which the synthetic polypeptide is added can be appropriately selected from the forms exemplified above. The aqueous cosmetic may contain a base other than water (such as the above-mentioned base), an active ingredient (such as the above-described active ingredient), and an additive (such as the above-described additive) as necessary. . The water-based cosmetic may contain other antibacterial components (such as the antibacterial components exemplified above), but usually the amount of other antibacterial agents is extremely small or substantially free of other antibacterial agents. In addition, antimicrobial properties can be effectively imparted by adding the synthetic polypeptide. The proportion of the other antibacterial agent in the aqueous cosmetic product can be selected from the range exemplified above.

  The amount of the synthetic polypeptide added is such that the concentration of the synthetic polypeptide in the resulting aqueous cosmetic is, for example, 0.01 to 10% by weight, preferably 0.01 to 5% by weight (for example, 0.05 to 3). % By weight), more preferably about 0.1 to 2% by weight.

  The application method to the outer skin of the aqueous cosmetic is not particularly limited, and can be appropriately applied to the outer skin according to the form of the cosmetic. For example, a liquid aqueous cosmetic product may be applied directly to the outer skin, or may be applied to the outer skin by impregnating a mask (a mask formed of a non-woven fabric or the like). Cosmetics may be applied by dripping or spraying. In addition, liquid aqueous cosmetics dissolve or mix (or dilute) solid cosmetics (solid cosmetics intended to be dissolved in skin lotions, for example, cosmetics in the form of capsules, sponges, films, etc.). And is suitable for applications where the resulting mixture is applied to the outer skin.

  The use of the aqueous cosmetic of the present invention is not particularly limited, and can be used for various cosmetics such as whitening cosmetics, moisturizing cosmetics, aging care cosmetics, sunburn cosmetics, acne cosmetics, and astringent cosmetics.

  The frequency | count of application of water-based cosmetics can be suitably selected according to a use etc., for example, can be applied to a predetermined site | part 1-5 times per day, Preferably about 1-3 times. In addition, water-based cosmetics are washed off with water or hot water as necessary after application to the outer skin depending on the type of other ingredients (active ingredients, other bases, additives, etc.) contained in the water-based cosmetics. It may be used without being washed away.

  The aqueous cosmetic product of the present invention is excellent in antibacterial properties and collagen-like properties, and thus is useful for application to not only the face but also the outer skin of the entire body such as limbs, back, bust and neck.

  Hereinafter, the present invention will be described in more detail based on examples, but the present invention is not limited to these examples.

Example 1
(1) Production of synthetic polypeptide 60 g of a peptide represented by the formula: Pro-Hyp-Gly (manufactured by Chisso Corporation) was dissolved in 1.2 L of phosphate buffer (pH 7.4) and stirred at room temperature. To the obtained mixed solution, 5.68 g of 1-hydroxybenzotriazole (condensation aid) was added, dissolved or suspended, and further cooled to 4 ° C. with stirring. To the obtained mixture, 201.6 g of 1-ethyl-3- (3-dimethylaminopropyl) carbodiimide hydrochloride (EDC.HCl) was added and stirred at 4 ° C. for 2 hours and then at 20 ° C. for 46 hours. did.

  A part of the obtained reaction solution was collected, diluted 60 times with water, and gel permeation chromatography (manufactured by GE Healthcare Biosciences, AKTA Explorer system, column: Superdex 200HR 10/30, flow rate) : 0.5 mL / min, eluent: 10 mM phosphate buffer (pH 7.4) containing 150 mM NaCl, and a polypeptide peak was observed at a molecular weight of 250,000 or more. The molecular weight was calculated by using Gel Filtration LMW Calibration Kit and Gel Filtration HMW Calibration Kit manufactured by GE Healthcare Biosciences as standard substances.

  The above reaction solution is pulverized with a mixer (BRAUN, Brown Multiquick 5550CA), and the pulverized product is diluted 6 to 8 times with water and stirred. Ultrafiltration membrane (Asahi Kasei Chemicals Co., Ltd., Microza Lab Module) AHP1010, membrane size: molecular weight cut off 50,000) was used for ultrafiltration. The filtrate containing 3 to 4 L of impurities was discharged by one filtration, diluted by adding water to the solution containing the target polypeptide, and further subjected to ultrafiltration. This operation was repeated 10 to 12 times, and a total of about 40 L was separated as a filtrate, and the remaining liquid was concentrated until the polypeptide concentration became slightly higher than about 1% by weight to obtain an aqueous polypeptide solution. The concentration of the aqueous polypeptide solution is calculated in the same manner as in Example 1. Based on the calculation result, an aqueous solution having a polypeptide concentration of 1% by weight (hereinafter simply referred to as “artificial collagen 1% aqueous solution”). ) Was prepared. The specific gravity of the aqueous solution was calculated as being the same as that of water. When a circular dichroism spectrum of a polypeptide was collected from a portion of a 1% artificial collagen solution, a positive cotton effect was observed at 225 nm and a negative cotton effect was observed at 198 nm, forming a triple helical structure. It was confirmed that

(2) Preparation of aqueous cosmetics 30 parts by weight of the artificial collagen 1% aqueous solution obtained in the above step (1) is diluted with 70 parts by weight of sterilized water, filled into a sealable container (sterilized), and heated to 60 ° C. Was then pasteurized for 30 minutes to prepare an aqueous cosmetic filled in a container (hereinafter sometimes referred to as 0.3% artificial collagen aqueous solution).

Test example 1 (general viable count test)
The artificial collagen 0.3% aqueous solution obtained in the step (2) of Example 1 was stored in an incubator (35 ° C.), and a viable cell count test was conducted every month. A follow-up test was conducted.

  More specifically, one container of artificial collagen 0.3% aqueous solution stored in an incubator is taken out every month, 1 mL of artificial collagen 0.3% aqueous solution is collected, and a general viable count test sheet (Chisso ( The number of viable bacteria on the sheet was measured after being dropped at 35 ° C. for 48 hours. The 0.3% artificial collagen aqueous solution was stored without opening the container until the viable cell count test was performed. As a result, the number of general viable bacteria was 0 in all from 1 month to 6 months, and no propagation of general viable bacteria was observed in an unopened state.

Test example 2
1% of the artificial collagen 0.3% aqueous solution obtained in step (2) of Example 1 is stored at room temperature, and the hand is washed with an antibacterial hand soap. The test was carried out twice a day, and after 1, 2, 4, 8, 16 and 32 days, the number of general bacteria, mold and yeast were tested.

  The number of general bacteria was determined by culturing at 30 ° C. for 72 hours by the SCDLP agar plate culture method (SCDLP medium: manufactured by Nippon Pharmaceutical Co., Ltd.), and measuring the number of viable bacteria in the sample. Calculated as the number of general bacteria. The number of molds and yeasts was measured by GPLP agar plate culture method (GPLP agar medium: manufactured by Nippon Pharmaceutical Co., Ltd.) for 7 days at 25 ° C., and the number of viable bacteria in the sample was measured. This was done by calculating the number in 1 g. The detection limit for general bacteria is 100. The results are shown in Table 1.

  As is clear from Table 1, the aqueous cosmetics of the examples are less than the detection limit for general bacteria, molds and yeasts even under open conditions, and no microbial growth is observed. Are better.

Test example 3 (loss test)
The 0.3% aqueous solution of artificial collagen obtained in the step (2) of Example 1 was evaluated in the decay test using the following isolates A to E.

(1) Test bacteria Isolate A: Escherichia coli NBRC 3972
Isolated B: Pseudomonas aeruginosa NBRC 13275
Isolate C: Staphylococcus aureus subsp. Aureus NBRC 13276
Isolate D: Candida albicans NBRC 1594
Isolated bacteria E: Aspergillus niger NBRC 9455
(2) Test 20 mL of an artificial collagen 0.3% aqueous solution (specimen) was placed in a sterilized synthetic resin container, and 0.1 mL of the test bacterial solution containing each of the test bacteria was added to and mixed with the sample. The obtained sample was preserve | saved at 25 degreeC +/- 1 degreeC, and it used for the following fluctuation test measurement after 1, 4, 7, 14, 21, 27, and 28 days.

  Among the above samples, for samples using isolates A to C, SCDLPA medium (SCDLP agar medium: manufactured by Nippon Pharmaceutical Co., Ltd.) is used at 32.5 ° C. ± 1 ° C., and isolates D and E are added. The sample used was cultured for 5 days at 22.5 ° C. ± 1 ° C. using a GPLPA medium (GPLP agar medium: manufactured by Nippon Pharmaceutical Co., Ltd.) by the pour plate culture method. It was measured as the number in 1 mL.

Note that the above bacterial solution using the isolated bacteria A to C was cultured for 18 to 24 hours at 32.5 ° C. ± 1 ° C. in the SCDA medium (SCD agar medium: manufactured by Eiken Chemical Co., Ltd.). After that, the cells were suspended in physiological saline and prepared so that the number of bacteria per mL was about 10 ⁵ . In addition, the above bacterial solution using the isolated bacteria D and E was cultured for 7 days at 22.5 ° C. ± 1 ° C. in a PDA medium (potato dextrose agar medium: manufactured by Eiken Chemical Co., Ltd.). Thereafter, the formed spores were suspended in physiological saline (containing 0.05 wt% polysorbate 80), and the number of spores per mL was adjusted to about 10 ⁵ .

  The results are shown in Table 2.

  As is apparent from the results in Table 2, the aqueous cosmetic products of the examples have high antibacterial properties against any of the above-mentioned isolates, and can remarkably suppress the propagation.

Claims (9)

  A cosmetic comprising a polypeptide, substantially free of antibacterial components, and having at least an amino acid sequence represented by the formula Pro-Y-Gly (where Y represents Pro or Hyp). And an aqueous cosmetic containing a synthetic polypeptide that forms a collagen-like structure.   The aqueous cosmetic according to claim 1, wherein the proportion of the antibacterial component is 100 ppm or less.   The aqueous cosmetic according to claim 1 or 2, wherein the proportion of the antibacterial component is 10 ppm or less.   It has a form of an aqueous solution or aqueous dispersion in which a synthetic polypeptide is dissolved or dispersed in water as a base, and the concentration of the synthetic polypeptide is 0.01 to 5% by weight. Water-based cosmetics according to the section.   The aqueous cosmetic product according to any one of claims 1 to 4, which has been sterilized.   The aqueous cosmetic according to any one of claims 1 to 5, wherein the synthetic polypeptide has a molecular weight peak in a range of 10,000 or more in the molecular weight distribution. The synthetic polypeptide is a polypeptide (I) composed of units represented by the following formulas (1) to (3), a peptide unit having an amino acid sequence represented by the following formula (4), and the following formula ( The aqueous cosmetic according to any one of claims 1 to 6, which is at least one polypeptide selected from polypeptides (II) comprising a peptide unit having the amino acid sequence represented by 5).
[-(OC- (CH ₂ ) _m -CO) _p- (Pro-Y-Gly) _n- ] _a (1)
[-(OC- (CH ₂ ) _m -CO) _q- (Z) _r- ] _b (2)
[-HN-R-NH-] _c (3)
(In the formula, m is an integer of 1 to 18, p and q are the same or different and 0 or 1, Y represents Pro or Hyp, and n represents an integer of 1 to 20. Z represents an amino acid residue or 2 to 2. Represents a peptide chain consisting of 10 amino acid residues, r represents an integer of 1 to 20, R represents a linear or branched alkylene group, and the ratio (molar ratio) between a and b is a / B = 100/0 to 30/70, c = 1 when p = 1 and q = 0, c = b when p = 0 and q = 1, p = 1 and q = 1 C = a + b when c = 0 and c = 0 when p = 0 and q = 0.)
-Pro-Y-Gly- (4)
(Where Y is the same as above)
-Pro-V-Gly-W-Ala-Gly- (5)
(In the formula, V represents Gln, Asn, Leu, Ile, Val or Ala, and W represents Ile or Leu.) A synthetic polypeptide is a peptide unit represented by the following formula:
-(Pro-Y-Gly) _n- (1a)
(In the formula, Y represents Pro or Hyp, and n represents an integer of 1 to 15)
The proportion of this unit is 80% by weight or more in the synthetic polypeptide, has a molecular weight peak in the range of 2 × 10 ^{4 to} 500 × 10 ^{4 in} the molecular weight distribution, and is synthesized. The aqueous cosmetic according to any one of claims 1 to 7, wherein at least a part of the polypeptide is capable of forming a triple helical structure.   To a water-based cosmetic, a synthetic polypeptide having an amino acid sequence represented by at least the formula Pro-Y-Gly (where Y represents Pro or Hyp) and forming a collagen-like structure is added. For suppressing the growth of microorganisms in the plant.