AU2022202825B1

AU2022202825B1 - Polypeptide, polypeptide composition and application, and prepared anti-wrinkle lotion thereof

Info

Publication number: AU2022202825B1
Application number: AU2022202825A
Authority: AU
Inventors: John Taylor TSUI
Original assignee: Australian Health Ind Co Pty Ltd
Current assignee: Australian Health Industry Co Pty Ltd
Priority date: 2022-04-28
Filing date: 2022-04-28
Publication date: 2023-09-07
Anticipated expiration: 2042-04-28
Also published as: CN114920801A; CN114920801B

Abstract

The present invention discloses a polypeptide and a polypeptide composition and an application, and a prepared anti-wrinkle lotion thereof, belonging to the field of biotechnology. The present invention includes a polypeptide 1 or a polypeptide 2; the polypeptide 1 has the following sequence: Ala-Glu-Lys-Glu-Met-Asp-Glu-Arg-Tyr (SEQ ID NO:1), and the polypeptide 2 has the following sequence: Trp-Thr-le-Val-Glu-His-Ile-Glu-Arg-Leu-Asn (SEQ ID NO:2). The two polypeptides of the present invention have a preferable effect of promoting the secretion of collagen; meanwhile, with smaller molecular weight, the two polypeptides can penetrate through stratum corneum freely to enhance the cell vitality of dermis, promote the cell proliferation, supplement and repair the lost collagen on the surface of skin such that subcutaneous tissues become strengthened to rapidly heal the damages (for example, from ultraviolet ray or oxidization), thus achieving the purposes of wrinkle removal and aging resistance. Moreover, the combination of the polypeptide 1 and the polypeptide 2 has a synergistic effect.

Description

Polypeptide, Polypeptide Composition and Application, and Prepared Anti-Wrinkle Lotion Thereof

Technical Field

The present invention relates to the field of biotechnology, and in particular to a polypeptide and a polypeptide composition and an application, and a prepared anti-wrinkle lotion thereof.

Background

The anti-wrinkle and anti-aging has been one of the hot topics in medical science, and the top priority in cosmetics industry. Based on the market survey of cosmetics in the United States, the moisturizing and anti-wrinkle products have accounted for more than half of the market shares in the commercially available cosmetics. With the increase of the proportion in the domestic aging population, the demand for moisturizing and anti-wrinkle skin care products is increasingly growing.

The appearance of wrinkle is the symbol of skin aging; skin aging is a part of aging of body; the aging of body is reflected in skin most obviously. Aging is a kind of natural law, but is influenced by other factors such as, living habit, ultraviolet ray, frequently exaggerated expression, lack of exercises, dry skin due to lack of water, environmental and psychological factors.

At present, there are various types of moisturizing and anti-wrinkle products in the cosmetics market. But the durable-slow release effect is far from the expected effect; moreover, the frequent use of such kind of product will cause burdens on skin and wastes of the active ingredients. Therefore, the exploration on the durable release of moisturizing and anti-wrinkle active ingredients becomes the key point to overcome the shortcomings of the existing moisturizing and anti-wrinkle products.

Any discussion of the prior art throughout the specification should in no way be considered as an admission that such prior art is widely known or forms part of the common general knowledge in the field.

It is an object of the present invention to overcome or ameliorate at least one of the disadvantages of the prior art, or to provide a useful alternative.

Unless the context clearly requires otherwise, throughout the description and the claims, the words "comprise", "comprising", and the like are to be construed in an inclusive sense as opposed to an exclusive or exhaustive sense; that is to say, in the sense of "including, but not limited to".

Summary

According to a first aspect, the present invention provides an isolated polypeptide, comprising a polypeptide 1, wherein the polypeptide 1 has the following sequence: Ala-Glu-Lys-Glu-Met Asp-Glu-Arg-Tyr (SEQ ID NO:1).

According to a second aspect, the present invention provides a polypeptide composition,

comprising the polypeptide 1 of the first aspect and a polypeptide 2, wherein the polypeptide

2 has the following sequence: Trp-Thr-le-Val-Glu-His-le-Glu-Arg-Leu-Asn (SEQ ID NO:2).

According to a third aspect, the present invention provides a composition, comprising the

polypeptide 1 of the first aspect and a polypeptide 2, and a dispersion carrier, wherein the

polypeptide 2 has the following sequence: Trp-Thr-le-Val-Glu-His-le-Glu-Arg-Leu-Asn (SEQ ID NO:2).

According to a fourth aspect, the present invention provides a composition, comprising the composition of the third aspect and at least an effective amount of an active pharmaceutical ingredient for treating skin damage.

According to a fifth aspect, the present invention provides a use of the polypeptide of the first aspect or the polypeptide composition of the second aspect in the preparation of a skin care product for external use.

According to a sixth aspect, the present invention provides an anti-wrinkle lotion, wherein the anti-wrinkle lotion is prepared by the following raw materials according to parts by weight: 0.5 1 parts of the polypeptide composition of claim 3, 2-5 parts of jojoba oil, 3-4 parts of isopropyl myristate, 2-4 parts of PEG-7 glyceryl cocoate, 10-15 parts of lecithin, 5-7 parts of carbomer, 5-7 parts of glycerin, 3-5 parts of propylene glycol, 1-2 parts of PEG-400, 0.5-1 parts of sodium hyaluronate, 0.5-1 parts of ceramide, 0.05-1 parts of citric acid and 30-60 parts of deionized water.

An aspect of the present invention is to provide a polypeptide, a polypeptide composition and an application, and a prepared anti-wrinkle lotion thereof. The polypeptides of the present invention have a preferable effect of promoting the secretion of collagen; meanwhile, with smaller molecular weight, the polypeptides can penetrate through stratum corneum freely to enhance the cell vitality of dermis, promote the cell proliferation, supplement and repair the

la lost collagen on the surface of skin such that subcutaneous tissues become strengthened to rapidly lb heal the damages (for example, from ultraviolet ray or oxidization), thus achieving the purposes of wrinkle removal and aging resistance. Moreover, the combination of the polypeptide 1 and the polypeptide 2 has a synergistic effect.

The technical solution of the present invention is achieved as follows:

The present invention provides a polypeptide, including a polypeptide 1 and a polypeptide 2; the polypeptide 1 has the following sequence: Ala-Glu-Lys-Glu-Met-Asp-Glu-Arg-Tyr (SEQ ID NO:1), and the polypeptide 2 has the following sequence: Trp-Thr-Ile-Val-Glu-His-le-Glu-Arg-Leu-Asn (SEQ ID NO:2).

The present invention further seeks to protect a polypeptide composition, including the above polypeptide 1 and polypeptide 2.

As a further improvement of the present invention, a mass ratio of the polypeptide 1 to the polypeptide 2 is (3-5):1.

The present invention further seeks to protect a polypeptide derivative, where the polypeptide derivative is a modified product of the above polypeptide 1 or polypeptide 2.

The present invention further seeks to protect a polypeptide derivative composition, where the polypeptide derivative composition includes modified products of the above polypeptide 1 and polypeptide 2 having a mass ratio of (2-7):1.

The present invention further protects a composition, including at least one of the above polypeptide 1 and polypeptide 2, a composition of at least one of the modified products of the above polypeptide 1 and polypeptide 2, and a dispersion carrier.

As a further improvement of the present invention, the dispersion carrier includes one or more of water, a buffer salt solution, a fatty alcohol, a solid wax, ethylene glycol, glycerin or an organic ester.

The present invention further seeks to protect a composition, including the above composition, and at least an effective amount of an active pharmaceutical ingredient for treating a certain disease.

The present invention further seeks to protect an application of the above polypeptide or the above polypeptide composition or the above polypeptide derivative or the above polypeptide derivative composition in the preparation of a skin care product for external use.

The present invention further seeks to protect an anti-wrinkle lotion, where the anti-wrinkle lotion is prepared by the following raw materials according to parts by weight: 0.5-1 parts of the above polypeptide composition, 2-5 parts of jojoba oil, 3-4 parts of isopropyl myristate, 2-4 parts of PEG-7 glyceryl cocoate, 10-15 parts of lecithin, 5-7 parts of carbomer, 5-7 parts of glycerin, 3-5 parts of propylene glycol, 1-2 parts of PEG-400, 0.5-1 parts of sodium hyaluronate, 0.5-1 parts of ceramide, 0.05-1 parts of citric acid and 30-60 parts of deionized water.

The present invention has one or more of the following beneficial effects: the present invention provides a novel polypeptide sequence and a polypeptide composition, including a polypeptide 1 and a polypeptide 2; such two polypeptides could produce better effect in promoting the secretion of collagen. Meanwhile, with smaller molecular weight, the two polypeptides can penetrate through stratum corneum freely to enhance the cell vitality of dermis, promote the cell proliferation, supplement and repair the lost collagen on the surface of skin such that subcutaneous tissues become strengthened to rapidly heal the damages (for example, from ultraviolet ray or oxidization), thus achieving the purposes of wrinkle removal and aging resistance. Moreover, the combination of the polypeptide 1 and the polypeptide 2 has a synergistic effect.

The anti-wrinkle lotion prepared in the present invention contains emollient ingredients, including glycerin, propylene glycol and PEG-400 and thus, has good spreadability on skin. The anti-wrinkle lotion forms a layer of film on the surface to achieve moistening and moisturizing effects. Meanwhile, sodium hyaluronate, ceramide and polypeptide composition are infiltrated into the inside of skin to be released slowly, thereby further exerting the moistening and moisturizing effects for a long time. The product of the present invention is steady and has long service life, simple preparation method and low production costs.

Detailed Description of the Embodiments

The technical solution in the examples of the present invention will be described clearly and integrally; obviously, the examples described herein are merely partial, but not all examples of the present invention. Based on the examples of the present invention, all the other examples obtained by a person skilled in the art without making inventive effort shall fall within the protection scope of the present invention.

The inventor of the present invention extracted active proteins from plants having cosmetic efficacy to obtain dozens of functional polypeptides by multi-step enzymolysis and screening.

An example of the present invention further provides a polypeptide, including a polypeptide 1 and a polypeptide 2; the polypeptide 1 has the following sequence: Ala-Glu-Lys-Glu-Met-Asp-Glu-Arg-Tyr (SEQ ID NO:1); the polypeptide 2 has the following sequence: Trp-Thr-le-Val-Glu-His-le-Glu-Arg-Leu-Asn (SEQ ID NO:2).

The polypeptide is a short peptide, and has a preferable effect of promoting the secretion of collagen and can be absorbed better. Meanwhile, with smaller molecular weight, the short peptide can penetrate through stratum corneum freely to enhance the cell vitality of dermis, promote the cell proliferation, supplement and repair the lost collagen on the surface of skin such that subcutaneous tissues become strengthened to rapidly heal the damages (for example, from ultraviolet ray or oxidization), thus achieving the purposes of wrinkle removal and aging resistance.

As commonly known by a person skilled in the art, in the above polypeptides, the amino acid represented by each abbreviation of the amino acid is show in Table 1:

Table 1

Name Symbol or abbreviation

Alanine A or Ala

Aspartic acid D or Asp

Arginine R or Arg

Asparagine N or Asn

Glutamic acid E or Glu

Lysine K or Lys

Methionine M or Met

Tyrosine Y or Tyr

Tryptophan W or Trp

Threonine T or Thr

Isoleucine I or lIle

Valine V or Val

Histidine H or His

Leucine L or Leu

A further example of the present invention provides a polypeptide composition, including a polypeptide 1 and a polypeptide 2 with a mass ratio of (3-5):1.

A further example of the present invention provides a polypeptide derivative, where the polypeptide derivative is a modified product based on the above polypeptide 1 or polypeptide 2.

The term "derivative" used herein refers to a variant of a reference molecule, e.g., a polypeptide. For example, the derivative of polypeptide may include a variant where one or two amino acids are substituted relative to the reference polypeptide, preferably, the derivative of polypeptide is a sequence derived from conservative substitution. The derivative may further relate to the polypeptide modified with or by, including but not limited to non-natural amino acids, D- amino acids, amino acids modified at amino and/or carboxyl terminals (N- or C-terminal), especially for the modifications of the amino at N-terminal and/or the carboxyl at C-terminal, fatty acid modifications, peptide mimics, pseudopeptides and other similar kinds of modified products.

As described herein, the polypeptide of the present invention may form polypeptide derivatives via modification. As described herein, the polypeptide of the present invention may form polypeptide derivatives via modification. The polypeptide of the present invention or derivatives thereof may be prepared by the method known by a person skilled in the art very well, including the well-known chemical synthesis method.

Specifically, the conservative variation sequence of the present invention refers to the sequence obtained by the conservative substitution of one or two amino acids, where the conservative substitution refers that a specified amino acid is substituted by another amino acid with similar properties. For example, examples which may be regarded as the conservative substitution (similar properties) include but not limited to:

a) alanine, serine and threonine

b) glutamic acid and aspartic acid;

c) asparagine and glutamine;

d) arginineand lysine;

e) isoleucine , leucine, methionine and; f) phenylalanine, tyrosine and tryptophan.

A further example of the present invention provides a polypeptide derivative composition, including modified products of the above polypeptide 1 and polypeptide 2 having a mass ratio of (2-7):1.

A further example of the present invention provides a composition, including at least one of the above polypeptide 1 and polypeptide 2, a composition of at least one of the modified products of the above polypeptide 1 and polypeptide 2, and a dispersion carrier. The dispersion carrier includes one or more of water, a buffer salt solution, a fatty alcohol, a solid wax, ethylene glycol, glycerin or an organic ester. The polypeptide is subjected to dispersion and liquidation by the dispersion carrier to be suitable for the field of cosmetics or skin care products especially, thus being capable of improving the contact adhesion and permeability thereof with skin.

A further example of the present invention provides a composition, including the above composition, and at least an effective amount of an active pharmaceutical ingredient for treating a certain disease.

The polypeptide has shorter chain segments by itself and thus, has better absorbable properties. The composition is in combination with the active pharmaceutical ingredients for treating skin damage to promote the exertion of the efficacy, thus improving the efficacy obviously.

A further example of the present invention further provides an application of the polypeptide or the polypeptide composition or the polypeptide derivative or the polypeptide derivative composition mentioned above in the preparation of a skin care product for external use.

Example 1

An anti-wrinkle lotion was prepared by the following raw materials according to parts by weight: 0.5 parts of polypeptide composition, 2 parts of jojoba oil, 3 parts of isopropyl myristate, 2 parts of PEG-7 glyceryl cocoate, 10 parts of lecithin, 5 parts of carbomer, 5 parts of glycerin, 3 parts of propylene glycol, 1 part of PEG-400, 0.5 parts of sodium hyaluronate, 0.5 parts of ceramide, 0.05 parts of citric acid and 30 parts of deionized water. The polypeptide composition included a polypeptide 1 and a polypeptide 2; and a mass ratio of the polypeptide 1 to the polypeptide 2 was 3:1.

Preparation method:

(1) lecithin, carbomer, glycerin, propylene glycol and PEG-400 were stirred and mixed, and subjected to ultrasonic treatment for 10 min at 300 W to form an emulsifier;

(2) the polypeptide composition, jojoba oil, isopropyl myristate, and PEG-7 glyceryl cocoate were mixed evenly, and heated up to 35°C to form an oil phase;

(3) sodium hyaluronate and ceramide were mixed with 1/3 deionized water evenly to form an aqueous phase;

(4) citric acid was added to the remaining deionized water to obtain a pH adjusting agent;

(5) the oil phase in the step (2) and the emulsifier in the step (1) were stirred and mixed, and dropwise added with the aqueous phase in the step (3), then added with the pH adjusting agent obtained in the step (4) at a stirring condition of 1000 r/min, and stirred continuously for 15h to obtain the anti-wrinkle lotion.

Example 2

An anti-wrinkle lotion was prepared by the following raw materials according to parts by weight: 1 part of polypeptide composition, 5 parts of jojoba oil, 4 parts of isopropyl myristate, 4 parts of PEG-7 glyceryl cocoate, 15 parts of lecithin, 7 parts of carbomer, 7 parts of glycerin, 5 parts of propylene glycol, 2 parts of PEG-400, 1 part of sodium hyaluronate, 1 part of ceramide, 1 part of citric acid and 60 parts of deionized water. The polypeptide composition included a polypeptide 1 and a polypeptide 2; and a mass ratio of the polypeptide 1 to the polypeptide 2 was 5:1.

The preparation method was substantially the same as that of Example 1.

Example 3

An anti-wrinkle lotion was prepared by the following raw materials according to parts by weight: 0.7 parts of polypeptide composition, 3.5 parts of jojoba oil and 3.5 parts of isopropyl myristate, 3 parts of PEG-7 glyceryl cocoate, 12 parts of lecithin, 6 parts of carbomer, 6 parts of glycerin, 4 parts of propylene glycol, 1.5 parts of PEG-400, 0.7 parts of sodium hyaluronate, 0.7 parts of ceramide, 0.2 parts of citric acid and 45 parts of deionized water. The polypeptide composition included a polypeptide 1 and a polypeptide 2; and a mass ratio of the polypeptide 1 to the polypeptide 2 was 4:1.

The preparation method was substantially the same as that of Example 1.

Comparative Example 1

Compared with Example 3, the polypeptide composition was replaced by polypeptide 1; and the others were the same.

Comparative Example 2

Compared with Example 3, the polypeptide composition was replaced by polypeptide 2; and the others were the same.

Comparative Example 3

Compared with Example 3, the polypeptide composition was replaced by an anti-wrinkle active ingredient, coenzyme Q10; and the others were the same.

Test Example 1: Stability test

30 mL of the anti-wrinkle lotions prepared in Examples 1-3 and Comparative Examples 1-3 were divided into 3 parts, each 10 ml was put to a sealed glass bottle, then respectively placed at 4°C, 25°C and 50°C for half a year, and sampling was performed respectively in January, March and June for observation; these lotions kept a steady state, free of a separated layer or precipitate.

Physical properties (including refractive index, pH value, viscosity, electrical conductivity and surface tension) of the anti-wrinkle lotions prepared in Examples 1-3 and Comparative Examples 1-3 were determined; the refractive index was determined by an Abbe refractometer; the pH value was determined by a pH meter; the viscosity was determined by a Brookfield viscometer; the electrical conductivity was determined by a conductometer; the surface tension was determined by a surface tension meter; all the samples were determined at 25°C and a humidity of 65%. The results are shown in Table 2.

Table 2

Group Refractive pH value Viscosity Electrical Surface tension index (nd) (mPa.s) conductivity (mN/m) (pS/cm)

Example 1 1.452 6.54 5.5 175.2 37.25

Example 2 1.455 6.47 5.3 174.7 36.78

Example 3 1.457 6.42 5.1 174.0 35.77

Comparati 1.324 6.21 6.7 193.5 40.21 ve

Example 1

Comparati 1.352 6.15 6.5 189.7 41.25 ve Example 2

Comparati 1.310 6.10 7.2 210.3 43.17 ve Example 3

It can be seen from Table 2 that the anti-wrinkle lotion of the present invention has good physical properties, a weak-acidity pH value close to a pH value of human skin, low viscosity and small surface tension, indicating that the anti-wrinkle lotion is easy to be spread out on skin.

Test Example 2: Effect test

60 women aged 36-52 were selected and divided into 6 groups, 10 women each group; the anti-wrinkle lotions prepared in Examples 1-3 and Comparative Examples 1-3 were applied to the women of the 6 groups continuously for 30 d on trial, thus performing comparison, analysis and evaluation on the anti-wrinkle effects of the anti-wrinkle lotions. The anti-wrinkle effects are the improvement conditions of facial wrinkles, eye wrinkles, forehead wrinkles, fine wrinkles and skin elasticity. The results are shown in Table 3.

Table 3

Group Improvement Improvement Improvement Improvement Improvement rate of facial rate of eye rate of rate of fine rate of skin wrinkles(%) wrinkles(%) forehead wrinkles(%) elasticity(%) wrinkles (%)

Slight Obvious Slight Obvious Slight Obvious Slight Obvious Slight Obvious

Example 1 30 70 30 70 40 60 10 90 40 60

Example 2 20 80 20 80 30 70 10 90 30 70

Example 3 20 80 20 80 20 80 0 100 20 80

Comparative 40 60 40 60 50 50 40 60 40 60 Example 1

Comparative 50 50 50 50 50 50 60 40 50 50 Example 2

Comparative 70 30 70 30 80 20 70 30 80 20 Example 3

It can be seen from the above table that the anti-wrinkle lotion prepared in the present invention has an obvious anti-wrinkle effect, and can obviously improve the facial wrinkles, eye wrinkles, forehead wrinkles and fine wrinkles and strengthen skin elasticity. Either of the two polypeptides of the present invention have a better effect in promoting the secretion of collagen; meanwhile, with smaller molecular weight, the two polypeptides can penetrate through stratum corneum freely to enhance the cell vitality of dermis, promote the cell proliferation, supplement and repair the lost collagen on the surface of skin, such that subcutaneous tissues become strengthened to rapidly heal the damages (for example, from ultraviolet ray or oxidization), thus achieving the purposes of wrinkle removal and aging resistance. Moreover, the combination of the polypeptide 1 and the polypeptide 2 could produce a synergistic effect.

What is described above are merely preferred examples of the present invention, but are not construed as limiting the present invention. Any modification, equivalent replacement, improvement and the like made within the spirit and principle of the present invention shall fall within the protection scope of the present invention.

SEQUENCE LISTING 28 Apr 2022

<110> AUSTRALIAN HEALTH INDUSTRY CO. PTY LTD

<120> Polypeptide, Polypeptide Composition and Application, and Prepared Anti-Wrinkle Lotion Thereof

<130> SG

<160> 2 2022202825

<170> PatentIn version 3.5

<210> 1 <211> 9 <212> PRT <213> Artificial Sequence

<220> <223> peptide 1

<400> 1

Ala Glu Lys Glu Met Asp Glu Arg Tyr 1 5

<210> 2 <211> 11 <212> PRT <213> Artificial Sequence

<220> <223> peptide 2

<400> 2

Trp Thr Ile Val Glu His Ile Glu Arg Leu Asn 1 5 10

Page 1

Claims

1. An isolated polypeptide, comprising a polypeptide 1, wherein the polypeptide 1 has the following sequence: Ala-Glu-Lys-Glu-Met-Asp-Glu-Arg-Tyr (SEQ ID NO:1).

2. A polypeptide composition, comprising the polypeptide 1 of claim 1 and a polypeptide 2, wherein the polypeptide 2 has the following sequence: Trp-Thr-le-Val-Glu-His-le-Glu-Arg Leu-Asn (SEQ ID NO:2).

3. The polypeptide composition according to claim 2, wherein a mass ratio of the polypeptide 1 to the polypeptide 2 is (3-5):1.

4. A composition, comprising the polypeptide 1 of claim 1 and a polypeptide 2, and a dispersion carrier, wherein the polypeptide 2 has the following sequence: Trp-Thr-le-Val-Glu-His-le Glu-Arg-Leu-Asn (SEQ ID NO:2).

5. The composition according to claim 4, wherein the dispersion carrier comprises one or more of water, a buffer salt solution, a fatty alcohol, a solid wax, ethylene glycol, glycerin or an organic ester.

6. A composition, comprising the composition of claim 4 and at least an effective amount of an active pharmaceutical ingredient for treating skin damage.

7. Use of the polypeptide of claim 1 or the polypeptide composition of claim 2 in the preparation of a skin care product for external use.

8. An anti-wrinkle lotion, wherein the anti-wrinkle lotion is prepared by the following raw materials according to parts by weight: 0.5-1 parts of the polypeptide composition of claim 3, 2-5 parts of jojoba oil, 3-4 parts of isopropyl myristate, 2-4 parts of PEG-7 glyceryl cocoate, 10-15 parts of lecithin, 5-7 parts of carbomer, 5-7 parts of glycerin, 3-5 parts of propylene glycol, 1-2 parts of PEG-400, 0.5-1 parts of sodium hyaluronate, 0.5-1 parts of ceramide, 0.05 1 parts of citric acid and 30-60 parts of deionized water.