TWI859182B - Peptide, method and composition for melanin polymerization and hair darkening - Google Patents

Abstract

An isolated peptide comprises a motif of CRRWQWR (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.

Description

Peptides, methods and compositions for melanin polymerization and hair darkening

The present invention relates to a peptide for melanin polymerization and hair darkening in mammals, and a method and composition thereof.

Melanin is a natural pigment that may be found in the skin, eyes, hair, feathers and scales, where it is responsible for coloring.

Melanin is characterized by an irregular polymer structure. There are two major types of melanin, including eumelanin, which is brown-black, and pheomelanin, which is reddish-yellow. Hair color is determined by the relative content of these two pigments.

Because melanin is an aggregate of smaller component molecules, the degree and distribution of melanin aggregation/polymerization will also affect hair color.

In the present invention, a peptide that effectively interacts with melanin and stimulates the polymerization of melanin is unexpectedly found, wherein the peptide contains a 7-amino acid motif (CRRWQWR) (SEQ ID NO: 1).

It is found that a peptide having such a motif can stimulate melanin polymerization. In some embodiments, the peptide for stimulating melanin polymerization of the present invention is selected from the group consisting of SEQ ID NOs: 4-14.

Therefore, in one aspect, the present invention provides an isolated peptide for increasing melanin polymerization. The isolated peptide contains a 7-amino acid motif as shown in SEQ ID NO: 1. In some specific embodiments, the isolated peptide for increasing melanin polymerization is selected from the group consisting of SEQ ID NOs: 4-14.

In another aspect, the present invention provides an isolated peptide for darkening hair color in a mammalian individual. The isolated peptide contains a 7-amino acid motif as shown in SEQ ID NO: 1. In some specific embodiments, the isolated peptide for darkening hair color is selected from the group consisting of SEQ ID NOs: 4-14.

In another aspect, the present invention provides a composition for darkening hair color in a mammalian subject. The composition comprises an effective amount of an isolated peptide, wherein the isolated peptide comprises a motif of 7 amino acids as shown in SEQ ID NO: 1. According to certain embodiments of the present invention, the composition may further comprise an acceptable carrier and may be formulated as a topical preparation. In some embodiments, the isolated peptide is selected from the group consisting of SEQ ID NOs: 4-14 and combinations thereof.

According to the present invention, the topical preparation may include an ointment, an aerosol, a lotion, a cream, a gel, a dropper, a spray, a liquid, a patch, a shampoo or a conditioner. In a preferred embodiment, the composition is formulated as a shampoo or a conditioner.

In another aspect, the present invention provides a method for darkening hair color in a subject in need thereof, the method comprising administering to the subject a composition of the present invention in an amount effective to stimulate polymerization of melanin in the subject. In a preferred embodiment of the present invention, the method is used to darken the hair color of the mammalian subject, preferably a human subject. In a preferred embodiment of the present invention, the composition is topically administered to the subject.

It should be understood that the above general description and the following detailed description are merely exemplary and illustrative and do not limit the present invention.

Unless otherwise defined herein, scientific and technical terms used herein have the meanings commonly understood by one of ordinary skill in the art.

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.

As used herein, the terms "darkening" or "darkening hair color" refer to darkening the appearance of hair, including, but not limited to, darkening natural hair color or restoring hair that has lost color due to aging (e.g., gray or white hair) or external insults (e.g., excessive exposure to sunlight or chlorine).

The term "peptide" is used herein in its conventional sense, i.e., a polymer in which the monomers are amino acids linked together by amide bonds, or a polypeptide. When the amino acid is an α-amino acid, either the L-optical isomer or the D-optical isomer may be used. In addition, it is also intended to include unnatural amino acids, such as β-alanine, phenylglycine, and homoarginine. Standard abbreviations for amino acids are used herein.

As used herein, the term "peptide for stimulating melanin polymerization" or "peptide for darkening hair color" refers to a peptide having a length of 7 or more amino acid residues and having substantial melanin polymerization activity. Preferably, the peptide stimulating melanin polymerization includes a peptide having up to about 50 amino acid residues or 40, or 30, or 25, or 20, or 15, or 10, or 6 amino acids.

As used herein, the term "individual" may refer to any animal classified as a mammal, including humans.

As used herein, the term "carrier" refers to a material commonly used in the formulation of pharmaceutical or cosmetic compositions 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 can be used, such as water, buffer, 0.8% saline, 0.3% glycine, hyaluronic acid and the like. The composition may contain physiologically acceptable auxiliary substances required to approximate physiological conditions, such as pH adjusters and buffers, tonicity adjusters, wetting agents and the like, for example, sodium acetate, sodium lactate, sodium chloride, potassium chloride, calcium chloride, sorbitan monolaurate, triethanolamine oleate, etc.

As used herein, the words "topical" or "topically" refer in their conventional sense to sites that may 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 refers to direct contact of the peptide with tissues such as skin or membranes containing melanin-producing cells.

In some specific embodiments, the present invention provides an isolated peptide that effectively interacts with melanin to stimulate the polymerization of melanin, wherein the peptide comprises a 7-amino acid motif (CRRWQWR) (SEQ ID NO: 1). Preferably, the peptide for stimulating melanin polymerization of the present invention is selected from the group consisting of SEQ ID NOs: 4-14 and combinations thereof.

In some embodiments, the present invention provides an isolated peptide for darkening hair color in a mammalian individual. The isolated peptide contains a 7-amino acid motif as shown in SEQ ID NO: 1. In some embodiments, the isolated peptide for darkening hair color is selected from the group consisting of SEQ ID NOs: 4-14 and combinations thereof.

In some embodiments, the present invention provides a composition for darkening hair color in a mammalian subject. The composition comprises an effective amount of an isolated peptide, wherein the isolated peptide comprises a motif of 7 amino acids as shown in SEQ ID NO: 1. According to certain embodiments of the present invention, the composition may further comprise an acceptable carrier and may be formulated as a topical preparation. In some embodiments, the isolated peptide is selected from the group consisting of SEQ ID NOs: 4-14 and combinations thereof.

In some embodiments, the topical preparation may include an ointment, an aerosol, a lotion, a cream, a gel, a dropper, a spray, a liquid, a patch, a shampoo or a conditioner. In a preferred embodiment, the composition is formulated as a shampoo or a conditioner.

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

The present invention provides the use of the isolated peptide containing a 7 amino acid motif as shown in SEQ ID NO: 1 as an active ingredient for various purposes. In a preferred embodiment, the isolated peptide of the present invention is combined with an acceptable carrier to form a topical preparation that can be placed on the skin. The topical preparation may include an ointment, lotion, paste, cream, gel, drop, suppository, spray, liquid, shampoo, conditioner, powder, and transdermal patch. Thickeners, diluents, emulsifiers, dispersing aids, or adhesives may be used as needed. Preferably, one function of the carrier is to enhance the skin penetration of the peptide of the present invention and should be able to deliver the peptide to melanocytes under in vivo conditions. Suitable carriers are known to those of ordinary skill in the art, including but not limited to water, dimethyl sulfoxide, ethanol, liposomes, liquid petrolatum, petrolatum dimethylformamide, 2-pyrrolidone, oleic acid, and Azone® brand penetration enhancers.

The present invention is further described by the following examples, which are provided for illustration and not limitation.

Embodiment

1. Sample preparation

All peptides (Table 1) were synthesized by Genomics Bioscience & Technology Co. Ltd. (Taipei, Taiwan), and the purity and composition of these peptides were determined by high performance liquid chromatography (HPLC) and mass spectrometry. Peptide stock solutions were prepared by dissolving 10 mg of freeze-dried peptide powder in 1 ml of double deionized water (ddH ₂ O) and then stored at -20 ^o C. Melanin was purchased from Sigma-Aldrich (St. Louis, MO, USA). Table 1. Synthetic peptide library Name sequence SEQ ID NO: 2 APRKNVRWCTISQ SEQ ID NO: 3 CTISQPEWFKCRR SEQ ID NO: 4 FKRW SEQ ID NO: 5 FKCRW 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 QWR SEQ ID NO: 13 CRRWQWRMKKL SEQ ID NO: 14 RRWQWRMKKL SEQ ID NO: 15 RMKKLGAPSI

2. Melanin polymerization

0.025 mg/ml melanin was treated with 50 μM peptide for 24 hours and then centrifuged at 10,000g for 5 minutes. The supernatant was removed from the tube, leaving only the melanin precipitate. After boiling in 1 M NaOH for 1 hour, the polymerization of melanin was quantified and the absorbance was measured at 400 nm by spectrophotometer.

result

As shown in Table 1, melanin treated with 50 μM peptide containing the CRRWQWR (SEQ ID NO: 1) motif showed obvious melanin polymerization. However, SEQ ID NOs: 2, 3, and 15 showed no effect on melanin polymerization (Figures 1 and 2).

Those skilled in the art will appreciate that changes may be made to the above specific embodiments without departing from the broad inventive concept of the present invention. Therefore, it should be understood that the present invention is not limited to the specific specific embodiments disclosed, but is intended to cover modifications within the spirit and scope of the present invention as defined by the appended claims.

without

When read in conjunction with the accompanying drawings, the above invention content and the following detailed description of the invention will be better understood. In order to illustrate the present invention, the currently preferred specific embodiments are shown in the accompanying drawings.

In the diagram:

Figure 1 shows the results of treating 0.025 mg/ml melanin with 50 μM peptide for 24 hours in a 96-well plate and then photographing it under an optical microscope.

Figure 2 shows the polymerization of melanin. 0.025 mg/ml melanin was treated with 50 μM peptide for 24 hours. Data are presented as mean ± SD from three independent experiments. ***p>0.001.

without

<110> Muscle Vitality Research Institute Co., Ltd.

<120> Peptides, methods and compositions for melanin polymerization and hair darkening

<130> 0766/REN0003TW

<160> 15

<170> PatentIn version 3.5

<210> 1

<211> 7

<212> PRT

<213> Artificial Sequence

<220>

<223> Peptide

<400> 1

<210> 2

<211> 13

<212> PRT

<213> Artificial Sequence

<220>

<223> Peptide

<400> 2

<210> 3

<211> 13

<212> PRT

<213> Artificial Sequence

<220>

<223> Peptide

<400> 3

<210> 4

<211> 8

<212> PRT

<213> Artificial Sequence

<220>

<223> Peptide

<400> 4

<210> 5

<211> 9

<212> PRT

<213> Artificial Sequence

<220>

<223> Peptide

<400> 5

<210> 6

<211> 11

<212> PRT

<213> Artificial Sequence

<220>

<223> Peptide

<400> 6

<210> 7

<211> 13

<212> PRT

<213> Artificial Sequence

<220>

<223> Peptide

<400> 7

<210> 8

<211> 18

<212> PRT

<213> Artificial Sequence

<220>

<223> Peptide

<400> 8

<210> 9

<211> 25

<212> PRT

<213> Artificial Sequence

<220>

<223> Peptide

<400> 9

<210> 10

<211> 11

<212> PRT

<213> Artificial Sequence

<220>

<223> Peptide

<400> 10

<210> 11

<211> 12

<212> PRT

<213> Artificial Sequence

<220>

<223> Peptide

<400> 11

<210> 12

<211> 7

<212> PRT

<213> Artificial Sequence

<220>

<223> Peptide

<400> 12

<210> 13

<211> 11

<212> PRT

<213> Artificial Sequence

<220>

<223> Peptide

<400> 13

<210> 14

<211> 10

<212> PRT

<213> Artificial Sequence

<220>

<223> Peptide

<400> 14

<210> 15

<211> 10

<212> PRT

<213> Artificial Sequence

<220>

<223> Peptide

<400> 15

Claims (7)

An isolated peptide for stimulating melanin polymerization, which is composed of the amino acids listed in SEQ ID NO: 11. The isolated peptide of claim 1 is used to prepare a composition for darkening hair color. A composition for darkening hair color, comprising an effective amount of an isolated peptide as described in claim 1 or 2; wherein the composition is formulated as a topical preparation in the form of an ointment, an aerosol, an emulsion, a cream, a gel, a drop, a spray, a liquid, a patch, a shampoo or a conditioner. The composition of claim 3 further comprises a physiologically acceptable carrier. The composition of claim 3, wherein the topical preparation is a shampoo. The composition of claim 3, wherein the topical preparation is a hair conditioner. A use of the isolated peptide as described in claim 1, which is used to prepare a topical preparation for darkening hair color, wherein the isolated peptide has the effect of stimulating melanin polymerization.