Polypeptide with anti-aging biological activity

Abstract

The invention relates to the field of biological medicines, in particular to a polypeptide with anti-aging biological activity and related application thereof. The polypeptide obtained in the invention has a definite anti-aging effect, and has no toxic and side effects due to natural sources.

Description

Technical Field

[0001] The present invention relates to the field of biomedicine, and in particular to a polypeptide with anti-aging biological activity and related uses thereof. Background Art

[0002] Aging is a universal natural physiological law of the human body, and there seems to be no effective way to avoid it. Skin aging is generally divided into endogenous aging determined by genes and exogenous aging caused by the environment. Endogenous aging and exogenous aging together lead to various manifestations of skin aging. The direction of aging itself is difficult to reverse, but the aging process can be delayed in various ways. Studies have shown that endogenous skin aging is mainly related to age, while exogenous skin aging is mainly related to environmental exposure.

[0003] Facial skin is the most susceptible to aging-related changes in humans. Due to the influence of internal and external factors, the skin gradually ages, resulting in significant changes in facial skin morphology, especially wrinkles and sagging. At present, the academic community has reached a consensus: the loss of collagen is a very important indicator of skin aging; when there is a lack of collagen in the skin, the dermis of the skin is prone to collapse, resulting in blurred jaw lines and sagging face, reflecting various signs of skin aging. There are many types of collagen, and the common types are type I, type II, type III, type V and type XI.

[0004] Type I collagen is the only component of collagen fibers in the dermis, accounting for more than 80% of the skin; it is the most abundant component in the dermal connective tissue. Collagen fibers are considered to be the dermal components that are most closely related to skin aging. In addition, type I collagen provides mechanical support in the human body, maintains the integrity of organs and the human body, and ensures the normal function of the human body. In addition to its biomechanical effects, it also has functions such as signal transduction, and the transportation of growth factors and cytokines.

[0005] Based on the defects of the prior art, the present application provides a polypeptide with anti-aging biological activity, which can effectively increase the content of various collagens in the skin, especially type I collagen, and achieve unexpected technical effects of anti-skin aging. Summary of the invention

[0006] The purpose of the present invention is to provide a polypeptide with anti-aging biological activity and related uses thereof.

[0007] The purpose of the present invention can be achieved by the following technical solutions:

[0008] The first aspect of the present invention relates to a polypeptide having anti-aging biological activity, the amino acid sequence of which is shown as: DTRILL.

[0009] In some embodiments, the polypeptide having anti-aging biological activity is acetylated at its amino terminus and amidated at its carboxyl terminus.

[0010] The second aspect of the present invention provides a method for preparing the biologically active polypeptide, which can be artificially synthesized by genetic engineering methods, directly obtained from cells by separation and purification methods, or more directly prepared by chemical synthesis.

[0011] The third aspect of the present invention provides the use of the polypeptide having anti-aging biological activity in the preparation of food, health products, medicine or cosmetics having anti-aging function.

[0012] In some embodiments, the present invention relates to the use of the polypeptide having anti-aging biological activity in preparing a composition for preventing and / or delaying skin aging.

[0013] The term "anti-aging" or "anti-aging" used in the present invention mainly refers to anti-skin aging. Specifically, it can be a concept that includes skin protection and improvement of skin condition, skin whitening, prevention or improvement of skin aging and wrinkles, pore contraction and reduction, skin protection, improvement of skin barrier function, reduction of skin irritation, skin cell proliferation and regeneration ability, antioxidant ability, and improvement of collagen synthesis ability, especially refers to the ability to improve collagen synthesis.

[0014] In some specific embodiments, the skin aging includes: loss of collagen, increased wrinkles, sagging skin, dullness, uneven skin tone, and dryness.

[0015] In some specific embodiments, the collagen includes: type I collagen, type II collagen, type III collagen, type V collagen, type IV collagen, type VII collagen, type XI collagen and type XVII collagen.

[0016] In a fourth aspect, the present invention provides a product comprising a derivative of the polypeptide having anti-aging biological activity; the derivative refers to a polypeptide derivative obtained by modifying the amino acid side chain group, amino terminus or carboxyl terminus of the biologically active polypeptide by hydroxylation, carboxylation, carbonylation, methylation, acetylation, phosphorylation, esterification or glycosylation, while not affecting its biological activity.

[0017] The fifth aspect of the present invention relates to a composition comprising the aforementioned polypeptide and a pharmaceutically, food or cosmetically acceptable carrier.

[0018] In some embodiments, the composition is selected from a pharmaceutical composition, a food composition, or a cosmetic composition.

[0019] In some embodiments, the dosage form of the composition includes an oral dosage form, a parenteral dosage form, or a topical dosage form.

[0020] In some embodiments, the dosage form of the composition further comprises a cream, an emulsion, an aqueous solution, a gel, an oil, a powder, a mud, a wax-based solution, a patch, a film or a freeze-dried solution.

[0021] In some preferred embodiments, the composition further comprises an excipient and an adjuvant.

[0022] In some embodiments, the excipients include: hydroxyethyl cellulose, hydroxypropyl cellulose, hydroxypropyl methyl cellulose, polyethylene glycol, polyacrylamide, polyacrylic acid, polyvinyl alcohol, dextran, lactic acid, citric acid, pyrrolidone carboxylic acid, glycerol, urea, sorbitol, amino acids, lanolin, petrolatum.

[0023] In some embodiments, the adjuvant includes: solubilizer, thickener, gelling agent, softener, antioxidant, suspending agent, stabilizer, foaming agent, flavoring agent, surfactant, water, ionic or non-ionic emulsifier, chelating agent, preservative, vitamin, retardant, wetting agent, essential oil, dye, pigment.

[0024] In the sixth aspect of the present invention, the bioactive peptides of the present application can be used in combination with other ingredients, including but not limited to: antioxidants, reactive carbonyl species scavengers, anti-glycation agents, antihistamines, antiviral agents, antiparasitic agents, emulsifiers, moisturizers, anti-wrinkle agents and / or anti-aging agents, etc.

[0025] In some specific embodiments, the anti-aging composition of the present invention may contain other antioxidants and the like.

[0026] In some preferred embodiments, the antioxidant includes but is not limited to: for example, citric acid, citric acid derivatives, vitamin C, vitamin C derivatives, lycopene, vitamin A, carotenoids, vitamin B, vitamin B derivatives, flavonoids, polyphenols, glutathione, selenium, sodium thiosulfate, vitamin E, vitamin E derivatives, pyrroloquinoline quinone derivatives, superoxide dismutase (SOD), glutathione peroxidase, glutathione-S-converter, glutathione reductase, catalase, ascorbate peroxidase, and mixtures thereof.

[0027] In the first to sixth aspects of the present invention, the concentration of the polypeptide having anti-aging biological activity is: 0.1-100 ppm, preferably 1-50 ppm, for example: 0.1 ppm, 1 ppm, 5 ppm, 10 ppm, 50 ppm, 100 ppm;

[0028] The inventors of the present application obtained a polypeptide DL-6 with potential anti-aging biological activity, whose amino acid sequence is DL-6 (DTRILL, modified in the form of acetylation at the amino terminus and amidation at the carboxyl terminus), which can significantly upregulate the secretion of type I collagen in HFF-1 cells after ultraviolet-induced damage, effectively reducing skin photoaging damage and delaying aging. Description of the drawings:

[0029] Figure 1 :The experimental results of the effect of DL-6 on the content of type I collagen in HFF-1 cells induced by ultraviolet. Example:

[0030] definition

[0031] Although the numerical ranges and parameter approximations shown in the broad scope of the present invention, the numerical values ​​shown in the specific embodiments are recorded as accurately as possible. However, any numerical value is bound to contain a certain error, which is caused by the standard deviation in their respective measurements. In addition, all ranges disclosed herein should be understood to cover any and all sub-ranges contained therein. For example, the range of "1 to 10" recorded should be considered to include any and all sub-ranges between the minimum value 1 and the maximum value 10 (including the endpoints); that is, all sub-ranges starting with a minimum value of 1 or greater, such as 1 to 6.1, and sub-ranges ending with a maximum value of 10 or less, such as 5.5 to 10. In addition, any reference referred to as "incorporated herein" should be understood to be incorporated in its entirety.

[0032] Amino acids referred to in this application are represented herein by standard three-letter or single-letter abbreviations.

[0033] Example 1: Anti-photoaging biological activity test

[0034] The specific experimental methods are as follows:

[0035] (1) Plating: HFF-1 cells (Fenghui Bio, CL0167) were digested and resuspended, and the number of cells was counted using a Counstar cell technology instrument. Finally, DMEN high-glucose medium (WISENT, WL006) containing cells (HFF-1 cells) was added to a 24-well plate. The final cell density was 8×10 4 After plating, the cells were placed in a cell culture incubator for 24 hours;

[0036] (2) Induction and sample addition: When the cell culture confluence is about 40% to 60%, the drug is administered. 1 mL of cell culture medium is added to each well of the blank control group and the model group; 1 mL of culture medium containing 100 ng / mL TGFβ1 is added to each well of the positive control group; 1 mL of culture medium containing the test substance at the corresponding concentration is added to each well of the sample group.

[0037] (3) UV induction: 24 h after administration, the model group (complete culture medium added after UVA irradiation), the positive control group (complete culture medium containing 100 ng / mL TGFβ1 added after UVA irradiation), and the sample group (complete culture medium containing 10 ppm and 50 ppm synthetic peptide added after UVA irradiation) received a total dose of 9 J / cm 2 At the same time, the blank control group was placed in the same environment (UVA radiation dose of 0 J / cm 2 ).

[0038] (4) Sample collection: After 24 hours of incubation, collect the cell culture supernatant in an EP tube and store it in a -80°C freezer. Add 0.5 mL of lysis buffer to each well and pipette to lyse. Collect the cell fluid in an EP tube, and store the collected samples in a -80°C freezer. Detect according to the type I collagen content detection kit.

[0039] The experimental results show (see Figure 1 ): After UVA irradiation, the type I collagen content in HFF-1 cells was significantly reduced. After being treated with 100ng / mL TGF-β1, 10ppm and 50ppm of DL-6, the type I collagen content was significantly increased. Compared with 50ppm of acEL-6 (sequence: ETRILL), peptide DL-6 showed a stronger ability to increase type I collagen. When its concentration was 10ppm and 50ppm, DL-6 upregulated the secretion of type I collagen in HFF-1 cells after UV-induced damage by 25.76% and 42.48%, respectively. Its effect was far better than that of acEL-6, achieving unexpected technical results.

[0040] The above description is only a preferred embodiment, which is only used as an example and does not limit the combination of features necessary to implement the present invention. The titles provided are not intended to limit the various embodiments of the present invention. Terms such as "comprising", "including" and "including" are not intended to be limiting. In addition, unless otherwise specified, plural forms are included when there is no numeral modification, and "or" means "and / or". Unless otherwise defined herein, all technical and scientific terms used herein have the same meaning as those commonly understood by those skilled in the art.

Claims

1. A polypeptide with anti-aging biological activity, whose amino acid sequence is as shown in: DTRILL.

2. The polypeptide with anti-aging biological activity according to claim 1, wherein the amino terminus is acetylated and the carboxyl terminus is amidated.

3. Use of the polypeptide with anti-aging biological activity according to any one of claims 1 to 2 in the preparation of a composition for preventing and / or delaying skin aging.

4. The use according to claim 3, characterized in that The skin aging includes: loss of collagen, increased wrinkles, sagging skin, dullness, uneven skin tone, and dryness.

5. The use according to claim 4, characterized in that The collagen includes: type I collagen, type II collagen, type III collagen, type V collagen, type IV collagen, type VII collagen, type XI collagen and type XVII collagen.

6. A composition, characterized in that Contains the polypeptide according to any one of claims 1 to 2, and a pharmaceutically, food or cosmetically acceptable carrier.

7. The composition according to claim 6, characterized in that The composition is selected from a pharmaceutical composition, a food composition or a cosmetic composition.

8. The composition according to claim 7, characterized in that The dosage form of the composition includes an oral dosage form, a parenteral dosage form, or an external dosage form.

9. The composition of any one of claims 6 to 8, further comprising an excipient and an adjuvant.

10. The composition of claim 9, wherein the excipient comprises: Hydroxyethylcellulose, hydroxypropylcellulose, hydroxypropylmethylcellulose, polyethylene glycol, polyacrylamide, polyacrylic acid, polyvinyl alcohol, dextran, lactic acid, citric acid, pyrrolidone carboxylic acid, glycerin, urea, sorbitol, amino acids, lanolin, petrolatum.

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