An oil-control polypeptide, its composition and uses

Abstract

Disclosed is an oil-control polypeptide, its composition and use, which relates to the technical field of polypeptides. The peptide or its salt has a structure shown in formula (I): R1-Val-Ser-Gly-Leu-Thr-Pro-R2 (I). The peptide or its salt, or their composition of the present disclosure has the functions of controlling oil, removing acne and repairing.

Description

Technical Field

[0001] The present disclosure relates to the technical field of polypeptides, in particular to an oil-control polypeptide and its composition, and their uses. Background Art

[0002] The sebaceous gland is a multi-acinar holocrine tissue composed of one or several sac-shaped acini and short ducts, distributed in most areas of the whole body skin, with the highest density in the scalp and face. Under normal physiological conditions, the sebaceous gland secretes an appropriate amount of oily waxy substances (sebum) to the surface of the stratum corneum, thus playing a role in nourishing, lubricating, inhibiting the growth and invasion of pathogenic microorganisms on the fur. However, some endogenous or exogenous factors, such as age, diet, temperature, humidity, ultraviolet rays or endocrine, etc., can cause hyperfunction of the sebaceous gland, resulting in excessive sebum secretion, so that the hair and skin surface appear greasy and shiny, and even cause skin problems such as pore blockage, blackheads or whitehead acne, and acne.

[0003] Human sebum is mainly composed of triglycerides, wax esters, free fatty acids, squalene and cholesterol, etc. Among them, the generation of triglycerides, wax esters and free fatty acids is closely related to the synthesis of fatty acids. Acetyl-CoA carboxylase (ACC) is a biotin enzyme isolated from the enzyme system for synthesizing long-chain fatty acids from acetyl-CoA, and plays an important role in the process of fatty acid synthesis. Acetyl-CoA carboxylase uses the energy provided by ATP in the organism to catalyze the conversion of acetyl-CoA into malonyl-CoA, and then malonyl-CoA, as a donor of two-carbon units, synthesizes fatty acids under the action of the fatty acid carbon chain elongation enzyme system. Therefore, inhibiting the activity or expression of acetyl-CoA carboxylase can limit the synthesis of fatty acids, inhibit the production of lipids in sebaceous cells, thereby alleviating sebum overflow and playing an oil-control role.

[0004] Currently, there are still few polypeptides with oil-control effects, and it is necessary to study more oil-control polypeptide compounds. Summary of the Invention

[0005] The present disclosure relates to a peptide, these peptides and the compositions containing these peptides, which have effects such as caring for or treating the skin, etc.

[0006] On the one hand, the present disclosure provides a peptide represented by formula (I), or its stereoisomer, or a mixture of its stereoisomers, or its salt,

[0007] R1-Val-Ser-Gly-Leu-Thr-Pro-R2 (I)

[0008] In formula (I),

[0009] R1 is selected from: H or R3-CO-, where R3 is selected from: substituted or unsubstituted alkyl, substituted or unsubstituted alkenyl;

[0010] R2 is selected from: -NR4R5 or -OR4, where each of R4 and R5 is independently selected from: H, substituted or unsubstituted alkyl, substituted or unsubstituted alkenyl;

[0011] The alkyl refers to a saturated aliphatic straight-chain or branched-chain alkyl having 1 to 24 carbon atoms (or having 1 to 16 carbon atoms; or having 1 to 14 carbon atoms; or having 1 to 12 carbon atoms; or having 1, 2, 3, 4, 5 or 6 carbon atoms); In some embodiments, the alkyl is selected from: methyl, ethyl, isopropyl, isobutyl, tert-butyl, pentyl, hexyl, heptyl, octyl, decyl, dodecyl, tetradecyl, hexadecyl, octadecyl, 2-ethylhexyl, 2-methylbutyl or 5-methylhexyl;

[0012] The alkenyl refers to a straight-chain or branched-chain alkenyl having 2 to 24 carbon atoms (or having 2 to 16 carbon atoms; or having 2 to 14 carbon atoms; or having 2 to 12 carbon atoms; or having 2, 3, 4, 5 or 6 carbon atoms); The alkenyl has one or more carbon-carbon double bonds, and in some embodiments, the alkenyl has 1, 2 or 3 conjugated or non-conjugated carbon-carbon double bonds; The alkenyl is bonded to the rest of the molecule through a single bond; In some embodiments, the alkenyl is selected from: vinyl, oleyl or linoleyl;

[0013] In some embodiments, the substituents in the "substituted alkyl" and "substituted alkenyl" are selected from C1-C4 alkyl; hydroxy; C1-C4 alkoxy; amino; C1-C4 aminoalkyl; C1-C4 carbonyloxy; C1-C4 oxycarbonyl; halogen (such as fluorine, chlorine, bromine, and iodine); cyano; nitro; azide; C1-C4 alkylsulfonyl; mercapto; C1-C4 alkylthio; C6-C 30 aryloxy such as phenoxy; -NR b (C=NR b )NR b R c where R b and R c are independently selected from: H, C1-C4 alkyl, C2-C4 alkenyl, C2-C4 alkynyl, C3-C 10 cycloalkyl, C6-C 18 aryl, C7-C 17 aralkyl, a heterocyclic group having three to ten members or a protecting group for amino.

[0014] In some embodiments, R1 is selected from: H, acetyl, tert-butyryl, hexanoyl, 2-methylhexanoyl, octanoyl, decanoyl, lauroyl, myristoyl, palmitoyl, stearoyl, oleoyl or linoleoyl; R4 and R5 are independently selected from: H, methyl, ethyl, hexyl, dodecyl or hexadecyl;

[0015] In some embodiments, R1 is selected from H, acetyl, lauroyl, myristoyl or palmitoyl; R4 is H and R5 is selected from: H, methyl, ethyl, hexyl, dodecyl or hexadecyl;

[0016] In some embodiments, R1 is H, acetyl, lauroyl, myristoyl or palmitoyl; R2 is -OH or -NH2.

[0017] In some embodiments, the peptide represented by formula (I), or its stereoisomer, or a mixture of its stereoisomers, or its salt, is selected from peptides (1)-(8):

[0018] (1) H-Val-Ser-Gly-Leu-Thr-Pro-NH2;

[0019] (2) H-Val-Ser-Gly-Leu-Thr-Pro-OH;

[0020] (3) Ac-Val-Ser-Gly-Leu-Thr-Pro-NH2;

[0021] (4) Ac-Val-Ser-Gly-Leu-Thr-Pro-OH;

[0022] (5) Pal-Val-Ser-Gly-Leu-Thr-Pro-NH2;

[0023] (6) Pal-Val-Ser-Gly-Leu-Thr-Pro-OH;

[0024] (7) Myr-Val-Ser-Gly-Leu-Thr-Pro-NH2;

[0025] (8) Myr-Val-Ser-Gly-Leu-Thr-Pro-OH.

[0026] The peptides of formula (I) of the present disclosure may exist as stereoisomers or mixtures of stereoisomers; for example, the amino acids they contain may have L-, D-configurations, or be racemic independently of each other. Thus, it is possible to obtain isomeric mixtures as well as racemic or diastereomeric mixtures, or pure diastereoisomers or enantiomers, depending on the number of asymmetric carbons and what isomeric or isomeric mixtures are present. In some embodiments, the structure of the peptides of formula (I) of the present disclosure is a pure isomer, i.e., an enantiomer or a diastereoisomer. In some embodiments, the structure of the peptides of formula (I) of the present disclosure is the L-isomer.

[0027] The present disclosure also includes all suitable isotopic variants of the peptides of formula (I). Isotopic variants of these peptides of the present disclosure are understood herein to mean compounds in which at least one atom within the peptides of the present disclosure is replaced by another atom of the same atomic number, but the atomic mass of the other atom is different from the atomic mass that is normally or predominantly present in nature. Examples of isotopes that can be incorporated into the peptides of the present disclosure are those of hydrogen, carbon, nitrogen or oxygen, such as 2 H (deuterium), 3 H (tritium), 13 C, 14 C, 15 N, 17 O or 18 O. Specific isotopic variants of the peptides of the present disclosure (especially those in which one or more radioactive isotopes have been incorporated) may be advantageous, for example, for examining the mechanism of action or the distribution of the active compound in the body; due to their relatively simple preparability and detectability, especially compounds labeled with 3 H or 14 C isotopes are suitable for this purpose. Additionally, due to the greater metabolic stability of the compound, the incorporation of an isotope (such as deuterium) can produce specific therapeutic benefits, such as an extended in vivo half-life or a reduced required active dose. The isotopic variants of the peptides of the present disclosure can be prepared by methods known to those skilled in the art, such as by the methods further described below and those described in the examples, by using the corresponding isotopically modified reagents and / or starting materials.

[0028] The term "salt" refers to a salt that is approved for use in animals, and more specifically in humans, and includes metal salts of the peptide of formula (I), where the metal includes, but is not limited to: lithium, sodium, potassium, calcium, magnesium, manganese, copper, zinc, or aluminum, etc.; includes salts formed by the peptide of formula (I) with organic bases, where the organic bases include, but are not limited to: ethylenediamine, ethanolamine, arginine, lysine, histidine, or piperazine, etc.; includes salts formed by the peptide of formula (I) with inorganic acids or organic acids, where the organic acids include, but are not limited to: acetic acid, citric acid, lactic acid, malonic acid, maleic acid, tartaric acid, fumaric acid, benzoic acid, aspartic acid, glutamic acid, succinic acid, oleic acid, trifluoroacetic acid, oxalic acid, pamoate, or gluconic acid, etc.; the inorganic acids include, but are not limited to: hydrochloric acid, sulfuric acid, boric acid, or carbonic acid.

[0029] The synthesis of the peptide of formula (I) or its salt according to the present disclosure can be carried out according to conventional methods known in the prior art, such as solid-phase synthesis, liquid-phase synthesis, or a method combining solid and liquid phases, and can also be prepared by biotechnological methods aimed at generating the desired sequence, or by controlled hydrolysis of proteins of animal, fungal, or plant origin.

[0030] For example, a method for obtaining the peptide of formula (I) includes the following steps:

[0031] - Coupling an amino acid with a protected N-terminus and a free C-terminus to an amino acid with a free N-terminus and a protected or solid-support-bound C-terminus;

[0032] - Removing the group protecting the N-terminus;

[0033] - Repeating the coupling sequence and removing the group protecting the N-terminus until the desired peptide sequence is obtained;

[0034] - Removing the group protecting the C-terminus or cleaving from the solid support.

[0035] In some embodiments, the C-terminus is bound to a solid support and the method is carried out on a solid phase, including coupling an amino acid with a protected N-terminus and a free C-terminus to an amino acid with a free N-terminus and a C-terminus bound to a polymer support; removing the group protecting the N-terminus; and repeating this sequence the required number of times so as to thereby obtain a peptide of the desired length, and then cleaving the synthesized peptide from the initial polymer support.

[0036] During the entire synthesis, the functional groups of the side chains of these amino acids are kept sufficiently protected with temporary or permanent protecting groups, and can be deprotected simultaneously or orthogonally with the process of cleaving the peptide from the polymer support.

[0037] In some embodiments, solid-phase synthesis can be carried out by a convergent strategy of coupling a dipeptide or tripeptide to a polymeric support or to a dipeptide or amino acid previously bound to the polymeric support.

[0038] Deprotecting the N-terminus and C-terminus and / or cleaving the peptide from the polymeric support in a non-determined order using standard conditions and methods known in the art, and subsequently the terminal functional groups can be modified. Optional modifications of the N-terminus and C-terminus can be carried out on the peptide bound to the polymeric support, or optional modifications of the N-terminus and C-terminus can be carried out after the peptide has been cleaved from the polymeric support.

[0039] Another aspect of the present disclosure provides a composition comprising an effective amount of the peptide represented by the above formula (I), or its stereoisomer, or a mixture of its stereoisomers, or its salt, and at least one excipient and an optional adjuvant.

[0040] In some embodiments, the adjuvant includes, but is not limited to: an agent that inhibits acetyl-CoA carboxylase activity, an analgesic, an agent that inhibits PAR-2 activity, a collagen synthesis stimulant, an agent that regulates PGC-1α synthesis, an agent that regulates the activity of PPARγ, an agent that increases or decreases the triglyceride content of adipocytes, an agent that stimulates or delays adipocyte differentiation, a lipolytic agent or an agent that stimulates lipolysis, a lipogenic agent, an inhibitor of acetylcholine receptor aggregation, an agent that inhibits muscle contraction, an anticholinergic agent, an elastase inhibitor, a matrix metalloproteinase inhibitor, a melanin synthesis stimulant or inhibitor, a whitening agent or a depigmenting agent, a pigmentation promoter, a self-tanning agent, a NO-synthase inhibitor, a 5α-reductase inhibitor, an inhibitor of lysyl hydroxylase and / or prolyl hydroxylase, an antioxidant, an agent against air pollution, an antiglycation agent, an antihistamine, an antiparasitic agent, an emollient, an organic solvent, a liquid propellant, a moisture-retaining substance, an α-hydroxy acid, a β-hydroxy acid, an epidermal hydrolase, a vitamin, an amino acid, a protein, a pigment, a biopolymer, a gum polymer, a thickening agent, a surfactant, an adhesive, a preservative, an anti-wrinkle agent, an agent capable of reducing or treating lower eye bags, a keratolytic agent, an antimicrobial agent, an agent that stimulates elastin synthesis, an agent that stimulates the synthesis of decorin, an agent that stimulates the synthesis of laminin, an agent that stimulates the synthesis of defensin, an agent that stimulates the synthesis of chaperone proteins, an agent that stimulates cAMP synthesis, an agent that stimulates hyaluronic acid synthesis, an agent that stimulates fibronectin synthesis, an agent that stimulates deacetylase synthesis, an agent that stimulates the synthesis of lipids and stratum corneum components, a ceramide, a fatty acid, an agent that inhibits collagen degradation, an agent that inhibits elastin degradation, an agent that inhibits serine protease, an agent that stimulates fibroblast proliferation, an agent that stimulates keratinocyte proliferation, an agent that stimulates adipocyte proliferation, an agent that stimulates melanocyte proliferation, an agent that stimulates keratinocyte differentiation, an agent that inhibits acetylcholinesterase, a skin relaxant, an agent that stimulates glycosaminoglycan synthesis, an anti-hyperkeratotic agent, an acne solvent, an anti-psoriatic agent, an anti-eczema agent, a DNA repair agent, a DNA protective agent, a stabilizer, an antipruritic agent, a curing agent, a tightening agent, a reconstructing agent, an agent that regulates sebum production, an antiperspirant, an agent that stimulates healing, an agent that assists in healing, an agent that stimulates re-epithelialization, an agent that assists in re-epithelialization, a cytokine, a sedative, an anti-inflammatory agent, an anesthetic, an agent that acts on capillary circulation and / or microcirculation, an agent that stimulates angiogenesis, an agent that inhibits vascular permeability, a venotonic agent, an agent that acts on cell metabolism, an agent for improving the dermal-epidermal junction, an agent that induces hair growth, a hair growth inhibitor or retardant, a fragrance, a chelating agent, a plant extract, an essential oil, a marine extract, an agent obtained from a biological fermentation process, an inorganic salt, a cell extract, and an organic or inorganic sunscreen agent that is effectively resistant to A and / or B ultraviolet rays or a mixture thereof.

[0041] The effective amount of the peptides of the present disclosure to be administered, as well as their dosages, will depend on many factors, including age, the condition of the user, the severity of the condition, the route and frequency of administration, and the specific nature of the peptides to be used.

[0042] "Effective amount" means an amount of one or more peptides of the present disclosure that is non-toxic but sufficient to provide the desired effect. The peptides of the present disclosure are used at effective concentrations in the compositions of the present disclosure to obtain the desired effect. In some embodiments, the concentration is between 0.00000001% (by weight) and 20% (by weight) relative to the total weight of the composition; in some embodiments, the concentration is between 0.000001% (by weight) and 15% (by weight) relative to the total weight of the composition; in some embodiments, the concentration is between 0.0001% (by weight) and 10% (by weight) relative to the total weight of the composition; in some embodiments, the concentration is between 0.0001% (by weight) and 5% (by weight) relative to the total weight of the composition.

[0043] Another aspect of the present disclosure provides a delivery system or sustained-release system to achieve better penetration of the active ingredient, which comprises an effective amount of the peptide represented by the above formula (I), or its stereoisomer, or a mixture of its stereoisomers, or its salt, or the above composition.

[0044] The term "delivery system" refers to a diluent, adjuvant, excipient or carrier administered together with the peptides of the present disclosure, which are selected from: water, oil or surfactants, including those of petroleum origin, animal origin, plant origin, or synthetic origin, such as and not limited to peanut oil, soybean oil, mineral oil, sesame oil, castor oil, polysorbates, sorbitan esters, ether sulfates, sulfates, betaines, glucosides, maltosides, fatty alcohols, nonoxynol, poloxamers, polyethylene oxides, polyethylene glycols, dextrans, glycerols, digitonin and the like. Those of ordinary skill in the art are aware of the diluents, adjuvants, excipients or carriers that can be used in different delivery systems in which the peptides of the present disclosure can be administered.

[0045] The term "sustained release" is used in its conventional meaning and refers to a delivery system of a compound that provides for the gradual release of the compound over a period of time. In some embodiments, the sustained-release system has a relatively constant level of compound release over the entire time period.

[0046] Examples of delivery systems or sustained-release systems include, but are not limited to: liposomes, oleosomes, ethosomes, millicapsules, microcapsules, nanocapsules, nanostructured lipid carriers, sponges, inclusion complexes, niosomes, micelles, milliballs, microspheres, nanospheres, lipid spheres, microemulsions, nanoemulsions, milliparticles, microparticles or nanoparticles.

[0047] Another aspect of the present disclosure provides a cosmetic comprising an effective amount of the peptide represented by the above formula (I), or its stereoisomer, or a mixture of its stereoisomers, or its salt, or the above composition, or the above delivery system or sustained-release system.

[0048] In some embodiments, the dosage form of the cosmetic includes paste, cream, emulsion, aqueous solution, oil, gel, powder, tablet, mud, patch, film, aerosol, spray, freeze-dried preparation or nano-preparation.

[0049] Another aspect of the present disclosure provides the use of the peptide represented by the above formula (I), or its stereoisomer, or a mixture of its stereoisomers, or its salt, or the above composition, or the above delivery system or sustained-release system in the preparation of a composition for skin care or treatment.

[0050] Another aspect of the present disclosure provides the use of the peptide represented by the above formula (I), or its stereoisomer, or a mixture of its stereoisomers, or its salt, or the above composition, or the above delivery system or sustained-release system in the preparation of a composition for oil control, acne treatment or repair.

[0051] Another aspect of the present disclosure provides the use of the peptide represented by the above formula (I), or its stereoisomer, or a mixture of its stereoisomers, or its salt, or the above composition, or the above delivery system or sustained-release system in the preparation of a composition for inhibiting skin oil synthesis or secretion, or in the preparation of a composition for reducing the content of acetyl-CoA carboxylase.

[0052] Another aspect of the present disclosure provides the use of the peptide represented by the above formula (I), or its stereoisomer, or a mixture of its stereoisomers, or its salt, or the above composition, or the above delivery system or sustained-release system in the preparation of a cosmetic.

[0053] In the present disclosure, the term "skin" should be understood as the multiple layers that make it up, from the outermost layer or stratum corneum to the innermost layer or subcutaneous tissue, including both endpoints. These layers are composed of different types of cells, such as keratinocytes, fibroblasts, melanocytes, and / or adipocytes, etc. In the present disclosure, the term "skin" includes the scalp.

[0054] The term "skin care" refers to maintaining and nourishing the skin, improving the state of the skin, and making the skin delicate, smooth, tender and healthy.

[0055] The present disclosure has the following advantages and effects:

[0056] The peptides of the present disclosure are obtained through artificial design, are convenient to synthesize, and have high safety. They can reduce the content of acetyl-CoA carboxylase, inhibit the synthesis or secretion of skin oil, have the effects of controlling oil, removing acne, and repairing, and can be used for skin care or treatment to improve skin problems. BRIEF DESCRIPTION OF THE DRAWINGS

[0057] In order to more clearly illustrate the technical solutions of the present disclosure, the drawings required for the description of the present disclosure will be briefly introduced below. Obviously, the drawings in the following description are only some embodiments of the present disclosure. For those of ordinary skill in the art, without creative efforts, other drawings can also be obtained based on these drawings.

[0058] Figure 1 It is the mass spectrometry diagram of the peptide (5) Pal-Val-Ser-Gly-Leu-Thr-Pro-NH2.

[0059] Figure 2 It is the mass spectrometry diagram of the peptide (6) Pal-Val-Ser-Gly-Leu-Thr-Pro-OH.

[0060] Figure 3 It is the result diagram of the effect of the test sample on the oil secretion of sebaceous gland cells.

[0061] Figure 4 It is the result diagram of the effect of the test sample on the content of acetyl-CoA carboxylase. DETAILED DESCRIPTION OF THE EMBODIMENTS

[0062] In order to make the above-mentioned objects, features, and advantages of the present disclosure more obvious and understandable, the present disclosure will be further described in detail below with reference to the drawings and embodiments. Obviously, the described embodiments are only a part of the embodiments of the present disclosure, rather than all the embodiments. All other embodiments obtained by those of ordinary skill in the art based on the embodiments of the present disclosure without creative efforts belong to the scope protected by the appended claims of the present disclosure.

[0063] In the present disclosure, the abbreviations for amino acids follow the rules specified by the IUPAC-IUB Commission of Biochemical Nomenclature in the European Journal of Biochemistry (Eur. J. Biochem. 1984, 138: 9-37).

[0064] Unless otherwise specified, the experimental reagents and materials used in the present disclosure can be obtained commercially. The following are the abbreviations for some reagents and materials:

[0065] Amide Resin: An initial resin for polypeptide synthesis (crosslinking degree 1%, substitution degree 1.72 mmol / g, particle size 100 - 200 mesh); 2-CTC Resin: An initial resin for polypeptide synthesis (2-chlorotrityl chloride resin); Fmoc-Linker: 4-[(2,4-dimethoxyphenyl)(Fmoc-amino)methyl]phenoxyacetic acid; Ac2O: Acetic anhydride; DMF: N,N-dimethylformamide; DIPEA: Diisopropylethylamine; DIC: Diisopropylcarbodiimide; piperidine: Piperidine; HOBt: 1-Hydroxybenzotriazole; DMAP: 4-Dimethylaminopyridine; Pal-OH: Palmitic acid; TFA: Trifluoroacetic acid; TIS: Triisopropylsilane; Val: Valine; Ser: Serine; Gly: Glycine; Leu: Leucine; Thr: Threonine; Pro: Proline; Fmoc: 9-Fluorenylmethoxycarbonyl; tBu: Tert-butyl.

[0066] Preparation of Example 1 Pal-Val-Ser-Gly-Leu-Thr-Pro-NH2

[0067] 1.1 Preparation of Fmoc-Linker-Amide Resin

[0068] Weigh 20 g of Amide Resin into a solid-phase synthesis reaction column, pour in 200 mL of DCM to swell for 30 min, wash the resin, and draw off the solvent.

[0069] Weigh 25 g of Fmoc-Linker and 7.5 g of HOBt into a dry Erlenmeyer flask. After dissolving with DMF solvent, place it at -20 °C for pre-freezing for 10 min, and add 10.7 mL of DIC for activation for 10 min.

[0070] Add the activated Fmoc-Linker to the swollen resin for reaction for 3 h, draw off the reaction solution, wash the resin, and draw off the solvent.

[0071] Continue to add Ac2O and DIPEA for capping treatment for 2 h. Wash the resin and draw off the solvent.

[0072] 1.2 Deprotect Fmoc

[0073] Deprotect Fmoc from Fmoc-Linker-Amide Resin twice with 20% piperidine / DMF, 10 min each time. Take a sample for Kaiser test, and the color shows dark blue. Wash the resin 7 times with DMF and draw off the solvent.

[0074] 1.3 Feed and React

[0075] Weigh 20.3 g of Fmoc-Pro-OH and 9.7 g of HOBt and add them to a dry Erlenmeyer flask. Add DMF to dissolve them, seal it, and place it in a -18°C refrigerator for 30 min. Add 13.9 mL of DIC to activate for 10 min, avoiding water vapor. Add the activated amino acid to the deprotected resin and react for 1 h, then draw off the reaction solution. The resin being colorless and transparent in the K test indicates that the reaction is complete.

[0076] Deprotect the N-terminal Fmoc group and couple the activated 23.9 g of Fmoc-Thr(tBu)-OH to the peptidyl resin using DMF as the solvent in the presence of 9.7 g of HOBt and 13.9 mL of DIC, and continue the reaction for 1 h. Then wash these resins and repeat the deprotection process of the Fmoc group to couple the next amino acid. In each coupling, in the presence of 9.7 g of HOBt and 13.9 mL of DIC, use DMF as the solvent and sequentially couple 21.3 g of Fmoc-Leu-OH, 17.8 g of Fmoc-Gly-OH, 23.0 g of Fmoc-Ser(tBu)-OH, and then 20.4 g of Fmoc-Val-OH; after the reaction is complete, wash the resin and draw off the solvent.

[0077] Deprotect the N-terminal Fmoc group of the peptidyl resin, deprotect Fmoc twice with 20% piperidine / DMF, 10 min each time, take a sample for the K test, and the color shows dark blue. Wash the resin 6 times with DMF and draw off the solvent.

[0078] In the presence of 16.2 g of HOBt and 23.1 mL of DIC, use DMF as the solvent and couple 25.7 g of Pal-OH to the peptidyl resin, continue the reaction for 1 h, wash the resin, draw off the solvent, and obtain 52.5 g of Pal-Val-Ser(tBu)-Gly-Leu-Thr(tBu)-Pro-Linker-Amide Resin after shrinkage drying.

[0079] 1.4 Cleavage

[0080] Measure 114 mL of TFA, 3 mL of TIS, and 3 mL of water, mix and stir evenly to obtain a cleavage solution, seal it, and place it in a -20°C refrigerator for standby; place isopropyl ether in a -20°C refrigerator for freezing and standby.

[0081] Weigh 20 g of Pal-Val-Ser(tBu)-Gly-Leu-Thr(tBu)-Pro-Linker-Amide Resin, add it to a round-bottom flask, add the above-prepared frozen cleavage solution, and stir for 2 h. Carry out suction filtration, collect the filtrate, concentrate it to 60 mL, then add 900 mL of isopropyl ether for sedimentation, stir and centrifuge with isopropyl ether for 6 times, and dry it in vacuo to obtain 7.5 g of crude peptide Pal-Val-Ser-Gly-Leu-Thr-Pro-NH2.

[0082] 1.5 Purification

[0083] Weigh 7.5 g of crude peptide Pal-Val-Ser-Gly-Leu-Thr-Pro-NH2 and dissolve it in acetic acid: methanol (V:V = 6:1) solution. After filtering through a microporous membrane with a pore size of 0.22 μm, add 10 mL of pure water to precipitate the gel, heat it to 50 °C to obtain a clear and transparent solution, load the sample, and purify it by reverse-phase HPLC. The purification gradient is as follows:

[0084]

[0085]

[0086] Inject for purification, collect the fractions, concentrate and freeze-dry to obtain peptide (5) Pal-Val-Ser-Gly-Leu-Thr-Pro-NH2 with a purity greater than 95%.

[0087] Example 2 Preparation of Pal-Val-Ser-Gly-Leu-Thr-Pro-OH

[0088] 2.1 Swelling of Resin

[0089] Weigh 20 g of 2-CTC Resin into a solid-phase synthesis reaction column, swell it with DCM, wash the resin, and draw off the solvent.

[0090] 2.2 Feeding Reaction

[0091] Weigh 16.0 g of Fmoc-Pro-OH and 5.9 g of HOBt into a dry Erlenmeyer flask. Dissolve them with DMF solvent, cool in an ice-water bath for 10 min, and activate with 8.4 mL of DIC for 10 min. Add the activated Fmoc-Pro-OH to the swollen resin and react for 3 h, draw off the reaction solution, wash the resin, and draw off the solvent. Continue to add DCM, MeOH, and DIPEA for capping treatment for 0.5 h. Wash the resin and draw off the solvent to obtain Fmoc-Pro-2-CTC Resin.

[0092] Deprotect the Fmoc group of Fmoc-Pro-2-CTC Resin twice with 20% piperidine / DMF for 10 minutes each time. Take samples for Kaiser test (K test), and the color shows dark blue. Wash the resin 7 times with DMF and remove the solvent by suction.

[0093] Weigh 14.4 g of Fmoc-Thr(tBu)-OH and 5.9 g of HOBt and add them to a dry Erlenmeyer flask. Add DMF to dissolve them, seal it and place it in a -18°C refrigerator for 30 minutes. Add 8.4 mL of DIC to activate for 10 minutes. Add the activated amino acid to the deprotected resin and react for 1.5 hours. Remove the reaction solution by suction. A colorless and transparent resin in the K test indicates that the reaction is complete.

[0094] Deprotect the N-terminal Fmoc group, and in the presence of 5.9 g of HOBt and 8.4 mL of DIC, use DMF as the solvent to couple 12.8 g of activated Fmoc-Leu-OH to the peptidyl resin, and continue the reaction for 1.5 hours. Then wash these resins and repeat the deprotection process of the Fmoc group to couple the next amino acid. In each coupling, in the presence of 6.9 g of HOBt and 9.8 mL of DIC, use DMF as the solvent to sequentially couple 12.6 g of Fmoc-Gly-OH, 16.1 g of Fmoc-Ser(tBu)-OH, and then 14.3 g of Fmoc-Val-OH; after the reaction is complete, wash the resin and remove the solvent by suction.

[0095] Deprotect the N-terminal Fmoc group of the peptidyl resin. Deprotect the Fmoc group twice with 20% piperidine / DMF for 10 minutes each time. Take samples for Kaiser test (K test), and the color shows dark blue. Wash the resin 7 times with DMF and remove the solvent by suction.

[0096] In the presence of 9.8 g of HOBt and 13.9 mL of DIC, use DMF as the solvent to couple 15.5 g of Pal-OH to the peptidyl resin, and continue the reaction for 1 hour. Wash the resin and remove the solvent by suction. After shrinking and drying, 39.5 g of Pal-Val-Ser(tBu)-Gly-Leu-Thr(tBu)-Pro-2-CTC Resin is obtained.

[0097] 2.3 Cleavage

[0098] Measure 209 mL of TFA, 5.5 mL of TIS and 5.5 mL of water, mix and stir evenly to obtain a cleavage solution. Seal it and place it in a -20°C refrigerator for later use.

[0099] Weigh 39.5 g of Pal-Val-Ser(tBu)-Gly-Leu-Thr(tBu)-Pro-2-CTC Resin, add it to a round-bottom flask, add the above-prepared frozen lysis solution, and stir for reaction for 2 h. Carry out suction filtration, collect the filtrate and concentrate it to 120 mL, add 1.2 L of pure water for sedimentation. After white solid precipitates, filter it with a Buchner funnel and wash it twice with pure water to obtain 13 g of crude peptide Pal-Val-Ser-Gly-Leu-Thr-Pro-OH.

[0100] 2.4 Purification

[0101] Weigh 13 g of crude peptide Pal-Val-Ser-Gly-Leu-Thr-Pro-OH, dissolve it in methanol and filter. First add a large amount of pure water to the filtrate, centrifuge once, then add 1% dilute ammonia water and centrifuge three times. Finally, add pure water and wash and centrifuge once more. Pre-freeze, concentrate and freeze-dry to obtain peptide (6) Pal-Val-Ser-Gly-Leu-Thr-Pro-OH with a purity greater than 95%.

[0102] Other peptides in formula (I) of the present invention can be prepared by a method similar to that in Example 1 and Example 2, and the molecular weights of these obtained peptides are determined by ESI-MS.

[0103] The test results of peptide (5) are shown in Figure 1 , and the results show that the mass-to-charge ratio (m / z) of the [M+H] + quasi-molecular ion peak is 810.80, and the molecular weight measured by mass spectrometry is 809.80.

[0104] The test results of peptide (6) are shown in Figure 2 , and the results show that the mass-to-charge ratio (m / z) of the [M+H] + quasi-molecular ion peak is 811.5451, and the molecular weight measured by mass spectrometry is 810.55.

[0105] Example 3 Oil Content Test

[0106] 3.1 Reagents and Materials

[0107] Trypsin digestion solution, DMEM medium, fetal bovine serum, phosphate buffer solution (PBS), palmitic acid and linoleic acid mixed solution (FFA), Oil Red O staining kit.

[0108] Among them, the preparation method of the palmitic acid and linoleic acid mixed solution (FFA) is as follows:

[0109] (1) Preparation of palmitic acid solution: Weigh 3.84 mg of palmitic acid, add 0.5 mL of 0.6 mg / mL NaOH solution, heat at 70 °C, and wait until it is completely dissolved. Then let it stand at room temperature until it solidifies. Subsequently, add 0.5 mL of isopropanol and continue heating until it dissolves again. Finally, add 1 mL of PBS to prepare a 2 mL system for standby.

[0110] (2) Preparation of linoleic acid solution: Weigh 4.2 mg of linoleic acid, add 0.5 mL of isopropanol, and then add 1.5 mL of PBS, heat to dissolve, and set aside for use.

[0111] (3) Preparation of FFA: Dilute the above linoleic acid solution and palmitic acid solution to 2250 μmol / L. After dilution, take 1 mL of each and mix well, then filter through a 0.22 μm microporous filter membrane to obtain it.

[0112] 3.2 Instruments

[0113] Microplate reader, CO2 incubator, laminar flow hood.

[0114] 3.3 Cell line

[0115] Sebaceous gland cells (SZ-95).

[0116] 3.4 Samples to be tested

[0117] Experimental group: Peptide (5), Peptide (6), with test concentrations of 5 ppm, 10 ppm, 25 ppm, 50 ppm.

[0118] Control group: PBS.

[0119] Model group: FFA.

[0120] 3.5 Experimental methods

[0121] Take a bottle of SZ-95 cells in the exponential growth phase with good condition, add 0.25% trypsin digestion solution, digest to make the adherent cells detached, count (1 - 4) × 10 5 cells / mL to make a cell suspension.

[0122] Take an appropriate amount of cell suspension and inoculate it on a 12-well plate, and place it in a constant temperature CO2 incubator for 24 h.

[0123] Except for adding PBS to the control group, add FFA to each well of the other wells to induce modeling, and then add different concentrations of samples to be tested to the experimental group respectively, and place them in a constant temperature CO2 incubator for 48 h.

[0124] Aspirate and discard the culture medium, operate according to the instructions of the Oil Red O staining kit, and calculate the relative content of cell lipid secretion.

[0125] 3.6 Experimental results

[0126] Oil Red O is a fat-soluble dye that can be highly dissolved in fat. Its staining principle is that Oil Red O can specifically adsorb neutral triglycerides, lipids, and lipoproteins in tissues and cells, thereby staining the fat. In this experiment, test samples were used to treat cells induced by FFA, and the amount of oil secretion in the corresponding cells was detected to determine whether the peptides of the present disclosure can inhibit oil secretion.

[0127] The results of the effect of the test sample on the oil secretion of sebaceous gland cells are shown in Figure 3 . The results showed that compared with the control group, the amount of oil secretion in the model group increased significantly, indicating that the modeling was successful. Compared with the model group, the experimental groups could significantly inhibit the oil secretion of sebaceous gland cells. It can be seen from this that the peptides of the present disclosure can inhibit skin oil secretion, slow down oil deposition, reduce the occurrence of acne or pimples, and also contribute to the recovery of the skin after the occurrence of acne or pimples, and have the effects of controlling oil, removing acne, and repairing.

[0128] Example 4 Measurement of the content of acetyl-CoA carboxylase

[0129] 4.1 Reagents and materials

[0130] Trypsin digestion solution, DMEM medium, fetal bovine serum, phosphate buffer (PBS), mixed solution of palmitic acid and linoleic acid (FFA), RIPA lysis buffer, BCA protein quantification kit, human acetyl-CoA carboxylase 1 ELISA detection kit. Among them, the preparation method of FFA is the same as that in Example 3.

[0131] 4.2 Instruments

[0132] Microplate reader, CO2 incubator, laminar flow hood, vortex mixer.

[0133] 4.3 Cell line

[0134] Sebaceous gland cells (SZ-95).

[0135] 4.4 Test samples

[0136] Experimental groups: Peptide (5), Peptide (6), test concentrations are 5 ppm and 10 ppm.

[0137] Control group: PBS.

[0138] Model group: FFA.

[0139] 4.5 Experimental method

[0140] Take a bottle of cells in the exponential growth phase with good condition, add 0.25% trypsin digestion solution, digest to make the adherent cells detached, count (1 - 4) × 10 5 cells / mL to make a cell suspension.

[0141] Inoculate the cell suspension onto a 12-well plate and culture it in a constant-temperature CO₂ incubator for 24 h.

[0142] Except for adding PBS to the control group, add FFA to each well of the remaining wells to induce modeling. Subsequently, add the test samples with different concentrations to the experimental groups respectively, and culture them in a constant-temperature CO₂ incubator for 48 h.

[0143] Collect the cells, centrifuge to obtain the cell precipitate, add RIPA lysis buffer, homogenize it 3 times with a vortex mixer (30 s / time, with an interval of 3 min), and centrifuge at 12000 rpm for 10 min. Aspirate the supernatant, operate according to the ELISA operation manual, and detect the total protein concentration of the supernatant with a BCA protein quantification kit, and calculate the content of acetyl-CoA carboxylase.

[0144] 4.6 Experimental results

[0145] Acetyl-CoA carboxylase is the rate-limiting enzyme for fatty acid synthesis. Inhibiting the activity or expression of acetyl-CoA carboxylase can inhibit fatty acid synthesis, reduce the sebum storage in sebaceous glands, and thus relieve seborrhea. In this experiment, cells induced by FFA were treated with the test samples, and the content of acetyl-CoA carboxylase in the corresponding cells was detected to determine whether the peptide of the present disclosure can inhibit the expression of acetyl-CoA carboxylase.

[0146] The results of the influence of the test samples on the content of acetyl-CoA carboxylase are shown in Figure 4 . The results show that compared with the control group, the content of acetyl-CoA carboxylase in the model group increased significantly. Compared with the model group, the experimental groups could all significantly reduce the content of acetyl-CoA carboxylase, indicating that the peptide of the present disclosure can inhibit the expression of acetyl-CoA carboxylase, thereby inhibiting the synthesis of oil.

[0147] It can be seen therefrom that the peptide of the present disclosure can reduce the content of acetyl-CoA carboxylase, inhibit the synthesis or secretion of skin oil, slow down the oil deposition, can reduce the occurrence of acne or pimples, and also helps the recovery of the skin after the occurrence of acne or pimples, and has the effects of controlling oil, removing acne, and repairing.

[0148] Example 5 Emulsion Containing Peptide (5)

[0149]

[0150]

[0151] Preparation method: According to the formula dosage, heat the materials in Phase A in an oil phase pot to 75 - 80 °C and stir until completely dissolved; add the materials in Phase B to an emulsifying pot, stir and heat to 80 - 85 °C until completely dissolved; pump Phase A into Phase B, turn on the vacuum, homogenize for 5 minutes, maintain stirring, and keep warm for 20 minutes; start cooling, cool to 60 - 65 °C, add the materials in Phase C and Phase D and continue stirring; cool to 35 - 40 °C, add the materials in Phase E, and stir for 10 - 15 minutes to obtain the product.

[0152] Example 6 Essence Containing Peptide (6)

[0153]

[0154] Preparation method: According to the formula dosage, add the materials in Phase A to a stirring pot and stir and heat to 80 - 85 °C; mix the materials in Phase B evenly until there are no powder particles, add them to the stirring pot, and continue stirring for 10 - 15 min; start cooling, cool to 60 - 65 °C, and add the materials in Phase C; cool to 35 - 40 °C, add the materials in Phase D and Phase E, and stir for 10 - 15 min to obtain the product.

[0155] Example 7 Cream Containing Peptide (1)

[0156]

[0157]

[0158] Preparation method: According to the formula dosage, heat the materials in Phase D in a suitable container to 55 - 60 °C until completely dissolved and set aside; add Phase A to a stirring pot and stir and heat to 80 - 85 °C; add the materials in Phase B to an oil phase pot, stir and heat to 75 - 80 °C until completely dissolved and transparent; put Phase B into Phase A, turn on the vacuum, homogenize for 5 min, maintain stirring, and keep warm for 20 min; start cooling, cool to 60 - 65 °C, add the materials in Phase C and the pre-dissolved Phase D materials, and homogenize for 2 min; cool to 35 - 40 °C, add the materials in Phase E, and stir for 10 - 15 min to obtain the product.

[0159] In the present disclosure, relational terms such as first and second are only used to distinguish one entity or operation from another entity or operation, and do not necessarily require or imply any actual relationship or order between these entities or operations. Moreover, the term "comprising", "including" or any other variation thereof is intended to cover non-exclusive inclusion, such that a process, method, article or terminal device comprising a series of elements includes not only those elements but also other elements not expressly listed, or elements inherent to such process, method, article or terminal device. Without further limitation, an element defined by the statement "comprising an..." does not exclude the presence of additional identical elements in the process, method, article or terminal device comprising the element.

[0160] Although specific embodiments of the present disclosure have been described for purposes of illustrative example, those skilled in the art can make various modifications or improvements without departing from the spirit and scope of the present disclosure. These modifications or improvements shall fall within the scope of the appended claims of the present disclosure.

Claims

1. The peptide represented by formula (I) or a salt thereof, R1-Val-Ser-Gly-Leu-Thr-Pro-R2 (I) In formula (I), R1 is selected from acetyl, lauroyl, myristoyl or palmitoyl; R2 is -OH or -NH2.

2. The peptide represented by formula (I) or a salt thereof according to claim 1, wherein, Selected from the following peptides (3)-(8): (3) Ac-Val-Ser-Gly-Leu-Thr-Pro-NH2; (4) Ac-Val-Ser-Gly-Leu-Thr-Pro-OH; (5) Pal-Val-Ser-Gly-Leu-Thr-Pro-NH2; (6) Pal-Val-Ser-Gly-Leu-Thr-Pro-OH; (7) Myr-Val-Ser-Gly-Leu-Thr-Pro-NH2; (8) Myr-Val-Ser-Gly-Leu-Thr-Pro-OH.

3. The peptide represented by formula (I) or a salt thereof according to claim 1 or 2, characterized in that the salt includes a metal salt of the peptide represented by formula (I), and the metal includes: lithium, sodium, potassium, calcium, magnesium, manganese, copper, zinc or aluminum; alternatively, the salt includes a salt formed by the peptide represented by formula (I) and an organic base, and the organic base includes: ethylenediamine, ethanolamine, arginine, lysine, histidine or piperazine; alternatively, the salt includes a salt formed by the peptide represented by formula (I) and an inorganic acid or an organic acid, and the organic acid includes: acetic acid, citric acid, lactic acid, malonic acid, maleic acid, tartaric acid, fumaric acid, benzoic acid, aspartic acid, glutamic acid, succinic acid, oleic acid, trifluoroacetic acid, oxalic acid, pamoic acid or gluconic acid; the inorganic acid includes: hydrochloric acid, sulfuric acid, boric acid or carbonic acid.

4. A composition, characterized in that, Comprising an effective amount of the peptide represented by formula (I) or a salt thereof according to any one of claims 1-3, and at least one excipient and an optional adjuvant.

5. A delivery system or sustained release system, characterized in that, Comprising an effective amount of the peptide represented by formula (I) or a salt thereof according to any one of claims 1-3, or the composition according to claim 4.

6. The delivery system or sustained release system according to claim 5, characterized in that, The delivery system or sustained-release system includes: liposomes, oleosomes, ethosomes, millimeter capsules, micron capsules, nanocapsules, sponges, inclusion compounds, niosomes, micelles, lipid spheres, micron emulsions, nanoemulsions, millimeter particles, micron particles or nanoparticles.

7. A cosmetic, characterized in that, Comprising an effective amount of the peptide represented by formula (I) or a salt thereof according to any one of claims 1-3, or the composition according to claim 4, or the delivery system or sustained-release system according to claim 5 or 6.

8. The cosmetic according to claim 7, characterized in that, The dosage form of the cosmetic includes paste, cream, emulsion, aqueous solution, oil, gel, powder, tablet, mud, patch, film, aerosol, spray, freeze-dried preparation or nano-preparation.

9. Use of the peptide represented by formula (I) or a salt thereof according to any one of claims 1-3, or the composition according to claim 4, or the delivery system or sustained-release system according to claim 5 or 6 in the preparation of a cosmetic composition for oil control, acne treatment or repair.

10. Use of the peptide represented by formula (I) according to any one of claims 1-3 or a salt thereof, or the composition according to claim 4, or the delivery system or sustained-release system according to claim 5 or 6 in the preparation of a composition for inhibiting skin oil synthesis or secretion.

11. The use according to claim 10, characterized in that, The inhibition of skin oil synthesis or secretion includes reducing the content of acetyl-CoA carboxylase.

12. Use of the peptide represented by formula (I) according to any one of claims 1-3 or a salt thereof, or the composition according to claim 4, or the delivery system or sustained-release system according to claim 5 or 6 in the preparation of a cosmetic for oil control, acne treatment or repair.

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