PEPTIDE, METHOD AND COMPOSITION FOR MELANIN POLYMERIZATION AND HAIR DARKENING

Abstract

An isolated peptide comprises a motif of RRWQW (SEQ ID NO: 1) for melanin polymerization and hair darkening is provided. Also disclosed herein are methods and compositions for hair darkening in a mammal subject.

Claims

1. An isolated peptide for stimulating melanin polymerization, comprising a fragment of SEQ ID NO: 1, and having up to 50 amino acids in length.

2. (canceled)

3. The isolated peptide of claim 1, wherein the peptide has up to 30 amino acids in length.

4. The isolated peptide of claim 1, wherein the isolated peptide is one selected from the group consisting of the peptides of SEQ ID NOs: 4-14.

5. An isolated peptide for hair darkening in a mammal subject, comprising a fragment of SEQ ID NO: 1, and having up to 50 amino acids in length.

6. (canceled)

7. The isolated peptide of claim 5, wherein the peptide has up to 30 amino acids in length.

8. The isolated peptide of claim 5, wherein the isolated peptide is one selected from the group consisting of the peptides of SEQ ID NOs: 4-14.

9. A composition for hair darkening in a mammal subject, comprising an effective amount of a peptide comprising a fragment of SEQ ID NO: 1, and having up to 50 amino acids in length, which is formulated as topical formulation in the form of an ointment, an aerosol, a lotion, a cream, a gel, drops, a spray, a liquid, a patch, a shampoo or a hair conditioner.

10. The composition of claim 9, wherein the peptide is one selected from the group consisting of the peptides of SEQ ID NOs: 4-14 and combinations thereof.

11. The composition of claim 10, further comprising a physiologically acceptable carrier.

12. The composition of claim 10, wherein the topical formulation is a shampoo.

13. The composition of claim 10, wherein the topical formulation is a hair conditioner.

14. A method for hair darkening in a subject in need thereof, comprising administering to the subject an effect amount of any one of the composition of claim 9 to stimulate melanin polymerization in the subject.

15. The method of claim 14, wherein the composition is administered to the subject topically.

Description

BRIEF DESCRIPTION OF THE SEVERAL VIEWS OF THE DRAWINGS

[0013] The patent or application file contains at least one color drawing. Copies of this patent or patent application publication with color drawing will be provided by the USPTO upon request and payment of the necessary fee.

[0014] The foregoing summary, as well as the following detailed description of the invention, will be better understood when read in conjunction with the appended drawings. For the purpose of illustrating the invention, there are shown in the drawings embodiments which are presently preferred.

[0015] In the drawings:

[0016] FIG. 1 shows that 0.025 mg/ml melanin was treated with 50 M peptides in a 96-well plate for 24 h, and then photographed under a light microscope.

[0017] FIG. 2 shows the polymerization of melanin. 0.025 mg/ml melanin was treated with 50 M peptide for 24 h. Data are represented as the meansSD from three independent experiments. ***p<0.001.

DETAILED DESCRIPTION OF THE INVENTION

[0018] Unless otherwise defined herein, scientific and technical terms used herein have the meanings that are commonly understood by those of ordinary skill in the art.

[0019] As used herein, the singular forms a, an, and the include plural referents unless the context clearly dictates otherwise. Thus, for example, reference to a sample includes a plurality of such samples and equivalents thereof known to those skilled in the art.

[0020] The term hair darkening or darkening the hair color as used herein refers to darkening the appearance of the hair, including, but not limited to, darkening natural hair color or restoring discolored hair due to aging (e.g., gray or white hair) or external aggressions (e.g., excess exposure to sun or chlorine).

[0021] The term peptide is used herein in its conventional sense, i.e., a polymer in which the monomers are amino acids and are joined together through amide bonds, alternatively referred to as a polypeptide. When the amino acids are -amino acids, either the L-optical isomer or the D-optical isomer may be used. Additionally, unnatural amino acids, for example, -alanine, phenylglycine and homoarginine are also meant to be included. Standard abbreviations for amino acids are used.

[0022] The term peptide for stimulating melanin polymerization or peptide for hair darkening as used herein refers to a peptide of 7 or more amino acid residues in length that has substantial melanin polymerization activity. Preferably, peptides up to about 50 amino acid residues or 40, or 30, or 25, or 20, or 15, or 10, or 6 amino acids are included in the peptide stimulating melanin polymerization.

[0023] The term subject as used herein can be any animal classified as a mammal, including a human.

[0024] The term carrier as used herein refers to materials commonly used on the formulation of pharmaceutical or cosmetic composition used to enhance stability, sterility and deliverability. When the peptide delivery system is formulated as a solution or suspension, the delivery system is in an acceptable carrier, preferably an aqueous carrier. A variety of aqueous carriers may be used, e.g., water, buffered water, 0.8% saline, 0.3% glycine, hyaluronic acid and the like. The compositions may contain physiologically acceptable auxiliary substances as required to approximate physiological conditions, such as pH adjusting and buffering agents, tonicity adjusting agents, wetting agents and the like, for example, sodium acetate, sodium lactate, sodium chloride, potassium chloride, calcium chloride, sorbitan monolaurate, triethanolamine oleate, etc.

[0025] The term topical or topically as used herein its conventional sense as referring to a spot which can be in or on any part of the body, including but not limited to the epidermis, any other dermis, or any other body tissue. Topical administration or application means the direct contact of the peptide with tissue, such as skin or membrane which contains melanin-producing cells.

[0026] In some embodiments, the present invention provides an isolated peptide effectively interacting with melanin, thereby stimulating the polymerization of melanin, wherein the peptide contains a 7-amino acid motif (CRRWQWR) (SEQ ID NO: 1). Preferably, the peptide for stimulating melanin polymerization of the present invent is selected from the group consisting of SEQ ID NOs: 4-14 and combinations thereof.

[0027] In some embodiments, the present invention provides an isolated peptide for hair darkening in a mammal subject. The isolated peptide contains a 7-amino acid motif as set forth in SEQ ID NO: 1. In some embodiments, the isolated peptide for hair darkening is selected from the group consisting of SEQ ID NOs: 4-14 and combinations thereof.

[0028] In some embodiments, the present invention provides a composition for hair darkening in a mammal subject. The composition comprises an effective amount of an isolated peptide contains a 7-amino acid motif as set forth in SEQ ID NO: 1. According to certain embodiments of the invention, the composition may further comprise an acceptable carrier, and may be formulated as a topical formulation. In some embodiments, the isolated peptide is selected from the group consisting of SEQ ID NOs: 4-14 and combinations thereof.

[0029] In some embodiments, the topical formulation may comprise an ointment, an aerosol, a lotion, a cream, a gel, drops, a spray, a liquid, a patch, a shampoo or a hair conditioner. In one preferred embodiment, the composition is formulated as a shampoo or a hair conditioner.

[0030] In some embodiments, the present invention provides a method for hair darkening in a subject in need thereof, which comprises administering to the subject the composition of the present invention in an amount effective to stimulate melanin polymerization in the subject. In preferred embodiments of the invention, the method is used to darken the hair color of the mammal subject, such as a human subject. In preferred embodiments of the invention, the composition is administered topically to the subject.

[0031] The present invention provides the use of the isolated peptide contains a 7-amino acid motif as set forth in SEQ ID NO: 1 as an active ingredient for various uses. In one preferred embodiment, the isolated peptide of the present invention is combined with an acceptable carrier to form a topical formulation which may be placed on the skin. Topical formulations may comprise an ointment, lotion, paste, cream, gel, drop, suppository, spray, liquid, shampoo, hair conditioner, powder and transdermal patch. Thickeners, diluents, emulsifiers, dispersing aids or binders may be used as needed. Preferably, one function of the carrier is to enhance skin penetration of the peptide of the present invention, and should be capable of delivering the peptide to melanocytes under in vivo conditions. Suitable carriers are well known to one of ordinary skill, and include but are not limited to water, dimethylsulfoxide, ethanol, liposome, liquid petrolatum, petrolatum dimethylformamide, 2-pyrrolidone, oleic acid, and Azone brand penetration enhancer.

[0032] The present invention is further illustrated by the following examples, which are provided for the purpose of demonstration rather than limitation.

Examples

[0033] 1. Sample Preparation

[0034] All peptides (Table 1) were synthesized by Genomics Bioscience & Technology co. Ltd (Taipei, Taiwan) and the purity and composition of these peptides was confirmed by high performance liquid chromatography (HPLC) and mass spectrometry. Peptide stocks were prepared by dissolving 10 mg of lyophilized peptide powder in 1 ml of double deionized water (ddH2O), and then stored at 20 C. Melanin was purchased from Sigma-Aldrich (St. Louis, Mo., USA).

TABLE-US-00001 TABLE 1 Library of synthesized peptides Name Sequence SEQ ID NO: 2 APRKNVRWCTISQ SEQ ID NO: 3 CTISQPEWFKCRR SEQ ID NO: 4 FKCRRWQW SEQ ID NO: 5 FKCRRWQWR SEQ ID NO: 6 FKCRRWQWRMK SEQ ID NO: 7 FKCRRWQWRMKKL SEQ ID NO: 8 FKCRRWQWRMKKLGAPSI SEQ ID NO: 9 FKCRRWQWRMKKLGAPSITCVRRAF SEQ ID NO: 10 KCRRWQWRMKK SEQ ID NO: 11 KCRRWQWRMKKL SEQ ID NO: 12 CRRWQWR SEQ ID NO: 13 CRRWQWRMKKL SEQ ID NO: 14 RRWQWRMKKL SEQ ID NO: 15 RMKKLGAPSI

[0035] 2. The Polymerization of Melanin

[0036] 0.025 mg/ml melanin were treated with 50 M peptides for 24 h, and then centrifuged at 10,000 g for 5 min. The supernatant was removed from the tube, leaving only the melanin pellet. The polymerization of melanin was quantified after boiling in 1 M NaOH for 1 hour and the absorbance was measured spectrophotometrically at 400 nm.

[0037] Results

[0038] Melanin treated with 50 M peptides, which contain CRRWQWR (SEQ ID NO: 1) motif as shown in Table 1, showed significantly melanin polymerization. However, SEQ ID NOs: 2, 3 and 15 did not show effects on melanin polymerization (FIGS. 1 and 2).

[0039] It will be appreciated by those skilled in the art that changes could be made to the embodiments described above without departing from the broad inventive concept thereof. It is understood, therefore, that this invention is not limited to the particular embodiments disclosed, but it is intended to cover modifications within the spirit and scope of the present invention as defined by the appended claims.