A polypeptide having an anti-aging activity

Abstract

The present application relates to the field of biological medicine, and particularly relates to a polypeptide with anti-aging activity and related use. The polypeptide obtained by the present application has clear anti-aging effect, and has no toxic side effects due to its natural source.

Description

TECHNICAL FIELD

[0001] The present application relates to the field of biological medicine, and in particular to a kind of polypeptide with anti-aging activity and its related use. BACKGROUND

[0002] Aging is a universal natural physiological law of human body, and there is currently no effective way to avoid it. Skin aging is generally divided into genetically determined endogenous aging and environmentally induced exogenous aging, and endogenous aging and exogenous aging together cause various manifestations of skin aging. The direction of aging itself is difficult to reverse, but the progress of aging can be delayed in various ways. Studies have shown that endogenous skin aging is mainly related to age, and exogenous skin aging is mainly related to environmental exposure.

[0003] Facial skin is the most susceptible to aging-related changes in human skin. Due to internal and external factors, the skin gradually ages, leading to significant changes in facial skin morphology, especially wrinkles and laxity. It has been agreed in the academic community that the loss of collagen is a very important indicator of skin aging; when the skin lacks collagen, the dermis layer of the skin is prone to collapse, leading to a blurred chin line, a sagging face, and a variety of 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% in the skin; it is the most abundant component in the dermal connective tissue. Collagen fibers are considered to be the most closely related dermal components 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 functioning of the human body. In addition to its biomechanical role, it also has functions such as signal transduction, transport of growth factors and cytokines, etc.

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

[0006] The purpose of the present application is to provide a kind of polypeptide with anti-aging activity and its related use.

[0007] The purpose of the present application can be realized by the following technical solutions:

[0008] In the first aspect of the present application, a kind of polypeptide with anti-aging activity, its amino acid sequence is as shown in formula (I): DTRILL.

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

[0010] In a second aspect of the present application, a method for preparing the bioactive polypeptide is provided. The bioactive polypeptide can be artificially synthesized by genetic engineering, directly obtained by isolation and purification from cells, or directly prepared by chemical synthesis.

[0011] In a third aspect of the present application, the polypeptide having anti-aging activity is provided for use in the preparation of a food, health product, medicine or cosmetic product having anti-aging function.

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

[0013] In the present application, the term "anti-aging" or "anti-aging" mainly refers to anti-skin aging, and specifically refers to a concept including all of 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 capacity, and improvement of collagen synthesis capacity, and particularly refers to the ability to improve collagen synthesis.

[0014] In some specific embodiments, the skin aging includes loss of collagen, increase in wrinkles, skin laxity, 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 of the present application, a product is provided, which comprises a derivative of the polypeptide having anti-aging activity. The derivative refers to a polypeptide derivative obtained by modification of the amino acid side chain group, the amino terminus or the carboxyl terminus of the bioactive polypeptide, such as hydroxylation, carboxylation, carbonylation, methylation, acetylation, phosphorylation, esterification or glycosylation, without affecting the biological activity.

[0017] In a fifth aspect of the present application, a composition is provided, which comprises the aforementioned polypeptide and a pharmaceutically or 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 comprises 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, a lotion, an aqueous solution, a gel, an oil, a powder, a paste, a wax-based, a patch, a film, or a lyophilized form.

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

[0022] In some embodiments, the excipient comprises 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 acid, lanolin, and petrolatum.

[0023] In some embodiments, the adjuvant comprises a solubilizer, a thickener, a gelling agent, a softener, an antioxidant, a suspending agent, a stabilizer, a foaming agent, a flavoring agent, a surfactant, an ionic or non-ionic emulsifier, a chelating agent, a preservative, a vitamin, a retarder, a humectant, an essential oil, a dye, and a pigment.

[0024] In a sixth aspect of the present application, the bioactive peptide of the present application can be used in combination with additional ingredients, including but not limited to, an antioxidant, a reactive carbonyl species scavenger, an anti-glycation agent, an antihistamine agent, an antiviral agent, an antiparasitic agent, an emulsifier, an emollient, an anti-wrinkle agent, and / or an anti-aging agent, etc.

[0025] In some specific embodiments, the anti-aging composition of the present application can comprise other antioxidants, etc.

[0026] In some preferred embodiments, the antioxidant comprises, but is not limited to, for example, citric acid, a citric acid derivative, vitamin C, a vitamin C derivative, lycopene, vitamin A, a carotenoid, vitamin B, a vitamin B derivative, a flavonoid, a polyphenol, glutathione, selenium, sodium thiothiocyanate, vitamin E, a vitamin E derivative, a pyrroloquinoline quinone derivative, superoxide dismutase (SOD), glutathione peroxidase, glutathione-S-transferase, glutathione reductase, catalase, ascorbate peroxidase, and a mixture of these, etc.

[0027] In the first to sixth aspects of the present application, the concentration of the polypeptide having anti-aging 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 this application obtained a polypeptide DL-6 with potential anti-aging bioactivity. Its amino acid sequence is DL-6 (DTRILL, modified by amino-terminal acetylation and carboxyl-terminal amidation). It can significantly upregulate the secretion of type I collagen in HFF-1 cells after UV-induced damage, effectively reducing skin photoaging damage and delaying aging. Attached Figure Description

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

[0030] definition

[0031] Although the numerical ranges and parameter approximations shown in the broad scope of this invention are intended to be approximated, the values ​​described in the specific embodiments are recorded as accurately as possible. However, any numerical value inherently contains a certain degree of error due to the standard deviation present in their respective measurements. Furthermore, all ranges disclosed herein should be understood to encompass any and all subranges contained therein. For example, the stated range “1 to 10” should be considered to include any and all subranges between the minimum value 1 and the maximum value 10 (inclusive); that is, all subranges beginning with a minimum value of 1 or greater, such as 1 to 6.1, and subranges ending with a maximum value of 10 or less, such as 5.5 to 10. Additionally, any references marked “incorporated herein” should be understood to be incorporated herein in their entirety.

[0032] The 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 method is as follows:

[0035] (1) Plate formation: After digestion and resuspending, HFF-1 cells (Fenghui Biotechnology, CL0167) were counted using a Council Cell Technology instrument. Finally, DMEN high-glucose medium (WISENT, WL006) containing HFF-1 cells was added to 24-well plates, achieving a final cell density of 8 × 10⁶ cells / well. 4 After plating, the cells were placed in a cell culture incubator and cultured 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; and 1 mL of culture medium containing the corresponding concentration of the test substance is added to each well of the sample group.

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

[0038] (4) Sample collection: After 24 h of incubation, the cell culture supernatant was collected in an EP tube and stored at -80°C. 0.5 mL of lysis solution was added to each well, and the cells were lysed by blowing. The cell lysate was collected in an EP tube, and the sample for detection was stored at -80°C. The sample was detected according to the type I collagen content detection kit.

[0039] The experimental results show (see Figure 1 ): After UVA irradiation, the content of type I collagen in HFF-1 cells was significantly reduced. After treatment with 100 ng / mL TGF-β1, 10 ppm, and 50 ppm of DL-6, the content of type I collagen was significantly increased. Compared with 50 ppm of acEL-6 (sequence: ETRILL), the peptide DL-6 showed stronger ability to increase type I collagen. When the concentration was 10 ppm and 50 ppm, the up-regulation rate of DL-6 on the secretion of type I collagen in HFF-1 cells after UV-induced damage was 25.76% and 42.48%, respectively, which was much better than acEL-6, and unexpected technical effects were achieved.

[0040] The above description is only a preferred embodiment, which is only an example and does not limit the combination of features necessary to implement the present application. The provided title is not intended to limit the various embodiments of the present application. The terms such as "contain", "include", and "comprise" are not intended to be limiting. In addition, unless otherwise specified, the plural form includes the singular form, and "or" means "and / or". Unless otherwise defined herein, all technical and scientific terms used herein have the same meaning as generally understood by those skilled in the art.

Claims

1. A polypeptide with anti-aging bioactivity, the amino acid sequence of which is shown as DTRILL.

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

3. The use of the polypeptide with anti-aging bioactivity according to any one of claims 1-2 in the preparation of compositions for preventing and / or delaying skin aging.

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

5. The use as described in claim 4, characterized in that, The collagen mentioned 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, It contains the polypeptide as described in any one of claims 1-2, and a pharmaceutically, food-, or cosmetically acceptable carrier.

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

8. The composition according to claim 7, characterized in that, The dosage forms of the composition include oral dosage forms, parenteral dosage forms, or topical dosage forms.

9. The composition according to any one of claims 6-8, further comprising excipients and adjuvants.

10. The composition of claim 9, wherein the excipient comprises: Hydroxyethyl cellulose, hydroxypropyl cellulose, hydroxypropyl methyl cellulose, polyethylene glycol, polyacrylamide, polyacrylic acid, polyvinyl alcohol, dextran, lactic acid, citric acid, pyrrolidone carboxylic acid, glycerin, urea, sorbitol, amino acids, lanolin, and mineral oil.

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