Oil control polypeptide as well as composition and application thereof

Abstract

The invention discloses an oil control polypeptide and a composition and application thereof, and relates to the technical field of polypeptides, and the peptide or a salt thereof has a structure as shown in a formula (I): R1-Val-Ser-Gly-Leu-Thr-Pro-R2 (I). The peptide or the salt of the peptide or the composition of the peptide and the salt has the effects of controlling oil, removing acnes and repairing.

Description

Technical Field

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

[0002] Sebaceous glands are multi-acinar holocrine secretory tissues composed of one or several cystic alveoli and short ducts. They are distributed in most areas of the skin, with the highest density on the scalp and face. Under normal physiological conditions, sebaceous glands secrete an appropriate amount of oily waxy substances (sebum) to the surface of the stratum corneum, thereby nourishing and lubricating the hair and inhibiting the growth and invasion of pathogenic microorganisms. However, some endogenous or exogenous factors, such as age, diet, temperature, humidity, ultraviolet rays or endocrine, can cause hyperfunction of sebaceous glands, resulting in vigorous sebum secretion, making the hair and skin surface greasy and shiny, and even causing skin problems such as clogged pores, blackheads or whiteheads, acne, etc.

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

[0004] At present, there are still few peptides with oil-controlling effects, and it is necessary to study more oil-controlling peptide compounds. Summary of the invention

[0005] The present disclosure relates to peptides, and the peptides and compositions containing the peptides have the effects of caring for or treating skin.

[0006] In one aspect, the present disclosure provides a peptide represented by formula (I), or a stereoisomer thereof, or a mixture of stereoisomers thereof, or a salt thereof,

[0007] R 1 -Val-Ser-Gly-Leu-Thr-Pro-R 2 (I)

[0008] In formula (I),

[0009] R1 Select from: H or R 3 -CO-, where R 3 Selected from: substituted or unsubstituted alkyl, substituted or unsubstituted alkenyl;

[0010] R 2 Selected from:-NR 4 R 5 OR 4 , where each R 4 and R 5 are independently selected from: H, substituted or unsubstituted alkyl, substituted or unsubstituted alkenyl;

[0011] The alkyl group refers to a saturated aliphatic linear or branched alkyl group having 1 to 24 carbon atoms (or 1 to 16 carbon atoms; or 1 to 14 carbon atoms; or 1 to 12 carbon atoms; or 1, 2, 3, 4, 5 or 6 carbon atoms); in some embodiments, the alkyl group 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 group refers to a straight or branched alkenyl group having 2-24 carbon atoms (or 2-16 carbon atoms; or 2-14 carbon atoms; or 2-12 carbon atoms; or 2, 3, 4, 5 or 6 carbon atoms); the alkenyl group has one or more carbon-carbon double bonds, and in some embodiments, the alkenyl group has 1, 2 or 3 conjugated or non-conjugated carbon-carbon double bonds; the alkenyl group is bonded to the rest of the molecule through a single bond; in some embodiments, the alkenyl group is selected from: vinyl, oleyl or linoleyl;

[0013] In some embodiments, the substituents in the "substituted alkyl" and "substituted alkenyl" are selected from C 1 -C 4 Alkyl; Hydroxyl; C 1 -C 4 Alkoxy; amino; C 1 -C 4 Aminoalkyl; C 1 -C 4 Carbonyloxy; C 1 -C 4 Oxycarbonyl; halogen (such as fluorine, chlorine, bromine, and iodine); cyano; nitro; azide; C 1 -C 4 Alkylsulfonyl; thiol; C 1 -C 4 Alkylthio; C 6 -C30 Aryloxy such as phenoxy; -NR b (C=NR b )NR b R c , where R b and R c are independently selected from: H, C 1 -C 4 Alkyl, C 2 -C 4 Alkenyl, C 2 -C 4 Alkynyl, C 3 -C 10 Cycloalkyl, C 6 -C 18 Aryl, C 7 -C 17 an aralkyl group, a heterocyclic group having three to ten members, or a protecting group for an amino group.

[0014] In some embodiments, R 1 is selected from: H, acetyl, tert-butyryl, hexanoyl, 2-methylhexanoyl, octanoyl, decanoyl, lauroyl, myristoyl, palmitoyl, stearoyl, oleoyl or linoleoyl; R 4 , R 5 are independently selected from: H, methyl, ethyl, hexyl, dodecyl or hexadecyl;

[0015] In some embodiments, R 1 is selected from H, acetyl, lauroyl, myristoyl or palmitoyl; R 4 is H and R 5 Selected from: H, methyl, ethyl, hexyl, dodecyl or hexadecyl;

[0016] In some embodiments, R 1 is H, acetyl, lauroyl, myristoyl or palmitoyl; R 2 Is -OH or -NH 2 .

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

[0018] (1)H-Val-Ser-Gly-Leu-Thr-Pro-NH 2 ;

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

[0020] (3)Ac-Val-Ser-Gly-Leu-Thr-Pro-NH 2 ;

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

[0022] (5)Pal-Val-Ser-Gly-Leu-Thr-Pro-NH 2 ;

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

[0024] (7)Myr-Val-Ser-Gly-Leu-Thr-Pro-NH 2 ;

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

[0026] The peptide shown in formula (I) of the present disclosure can exist as a mixture of stereoisomers or stereoisomers; for example, the amino acids contained therein can have L-, D- configurations, or be racemic independently of each other. Therefore, it is possible to obtain isomeric mixtures and racemic mixtures or diastereomeric mixtures, or pure diastereomers or enantiomers, depending on the number of asymmetric carbons and what isomers or isomeric mixtures are present. In some embodiments, the structure of the peptide shown in formula (I) of the present disclosure is a pure isomer, that is, an enantiomer or a diastereomer. In some embodiments, the structure of the peptide shown in formula (I) of the present disclosure is an 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 refer to compounds in which at least one atom in 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 usually 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, e.g. 2 H (deuterium), 3 H (tritium), 13 C. 14 C. 15 N. 17 O or 18 O. Certain isotopic variants of the peptides disclosed herein (particularly those into which one or more radioactive isotopes have been incorporated) may be useful, for example, to examine the mechanism of action or distribution of the active compound in vivo; due to the relative ease of preparation and detectability, particularly with3 H or 14 Compounds labeled with C isotopes are suitable for this purpose. In addition, due to the greater metabolic stability of the compound, the incorporation of isotopes (e.g., deuterium) can produce specific therapeutic benefits, such as an extension of the half-life in vivo or a reduction in the required active dose. 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 in the examples, by using the respective reagents and / or corresponding isotopic modifications of the starting materials.

[0028] The term "salt" refers to a salt approved for use in animals, and more specifically in humans, including metal salts of the peptide represented by formula (I), the metal including, but not limited to, lithium, sodium, potassium, calcium, magnesium, manganese, copper, zinc or aluminum, etc.; including salts formed between the peptide represented by formula (I) and an organic base, the organic base including, but not limited to, ethylamine, diethylamine, ethylenediamine, ethanolamine, diethanolamine, arginine, lysine, histidine or piperazine, etc.; including salts formed between the peptide represented by formula (I) and an inorganic acid or an organic acid, the organic acid including, but 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 acid including, but not limited to, hydrochloric acid, sulfuric acid, boric acid or carbonic acid.

[0029] The synthesis of the peptide represented by formula (I) or its salt can be carried out according to conventional methods known in the prior art, such as solid phase synthesis, liquid phase synthesis or a combination of solid phase and liquid phase. It can also be prepared by biotechnology methods aimed at producing the desired sequence, or by controlled hydrolysis of proteins of animal, fungal or plant origin.

[0030] For example, a method for obtaining a peptide represented by formula (I) comprises the following steps:

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

[0032] - Elimination of the group protecting the N-terminus;

[0033] - repeating this coupling sequence and eliminating the group protecting the N-terminus until the desired peptide sequence is obtained;

[0034] - Elimination of the group protecting the C-terminus or cleavage 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, comprising coupling an amino acid having a protected N-terminus and a free C-terminus with an amino acid having a free N-terminus and a C-terminus bound to a polymer support; eliminating the group protecting the N-terminus; and repeating this sequence as many times as required to thereby obtain a peptide of the desired length, followed by cleavage of the synthesized peptide from the initial polymer support.

[0036] The functional groups of the amino acid side chains remain fully protected with temporary or permanent protecting groups throughout the synthesis and can be deprotected simultaneously or orthogonally to the process of cleavage of the peptide from the polymer support.

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

[0038] The functional groups at the termini may be modified by deprotecting the N-terminus and C-terminus and / or cleaving the peptide from the polymer support in an undefined order using standard conditions and methods known in the art. The optional modification of the N-terminus and C-terminus may be performed on the peptide bound to the polymer support, or after the peptide has been cleaved from the polymer 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 a stereoisomer thereof, or a mixture of stereoisomers thereof, or a salt thereof, 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 the activity of acetyl-CoA carboxylase, an analgesic, an agent that inhibits the activity of PAR-2, a collagen synthesis stimulator, an agent that regulates the synthesis of PGC-1α, 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 fat decomposition, an adipogenic 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 stimulator or inhibitor, a whitening agent or a depigmenting agent, a pigmentation promoting agent, a self-tanning agent, a NO-synthase inhibitor, a 5α-reduct ... proenzyme inhibitors, inhibitors of lysyl hydroxylase and / or prolyl hydroxylase, antioxidants, agents against air pollution, anti-glycation agents, antihistamines, antiparasitic agents, emollients, organic solvents, liquid propellants, moisture retaining substances, alpha hydroxy acids, beta hydroxy acids, epidermal hydrolases, vitamins, amino acids, proteins, pigments, biopolymers, gelling polymers, thickeners, surfactants, adhesives, preservatives, anti-wrinkle agents, agents capable of reducing or treating under-eye bags, keratolytic agents, antimicrobial agents, agents stimulating elastin synthesis, agents stimulating decorin synthesis, agents stimulating laminin synthesis, agents stimulating defensin synthesis, agents stimulating chaperone protein synthesis, agents stimulating cAMP agents that stimulate synthesis of hyaluronic acid, agents that stimulate synthesis of fibronectin, agents that stimulate synthesis of deacetylases, agents that stimulate synthesis of lipids and stratum corneum components, ceramides, fatty acids, agents that inhibit collagen degradation, agents that inhibit elastin degradation, agents that inhibit serine proteases, agents that stimulate fibroblast proliferation, agents that stimulate keratinocyte proliferation, agents that stimulate adipocyte proliferation, agents that stimulate melanocyte proliferation, agents that stimulate keratinocyte differentiation, agents that inhibit acetylcholinesterase, skin relaxants, agents that stimulate glycosaminoglycan synthesis, anti-hyperkeratosis agents, comedolytic agents, anti-psoriatic agents, anti-eczema agents, DNA repair agents, DNA protective agents, stabilizers, antipruritic agents, solid agents, astringents, restructurers, agents regulating sebum production, antiperspirants, agents stimulating healing, agents assisting healing, agents stimulating re-epithelialization, agents assisting re-epithelialization, cytokines, sedatives, anti-inflammatory agents, anesthetics, agents acting on capillary circulation and / or microcirculation, agents stimulating angiogenesis, agents inhibiting vascular permeability, agents of venous tension, agents acting on cellular metabolism, agents for improving dermal-epidermal junction, agents inducing hair growth, agents inhibiting or retarding hair growth, fragrances, chelating agents, plant extracts, essential oils, marine extracts, agents obtained from biological fermentation processes, inorganic salts, cell extracts, and organic or inorganic photoprotective agents effective against A and / or B ultraviolet rays, or mixtures thereof.

[0041] The effective amount of the peptides of the present disclosure to be administered, as well as their dosage, will depend on a number of factors, including the age, state of the user, the severity of the condition, the route and frequency of administration, and the specific nature of the peptide 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 in the compositions of the present disclosure at effective concentrations 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 a salt thereof, or the above composition.

[0044] The term "delivery system" refers to a diluent, adjuvant, excipient or carrier used together with the peptide of the present disclosure, which is selected from: water, oil or surfactant, 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, polysorbate, sorbitan esters, ether sulfates, sulfates, betaines, glucosides, maltosides, fatty alcohols, nonoxynol ethers, poloxamers, polyoxyethylene, polyethylene glycol, dextrose, glycerol, 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 peptide of the present disclosure can be administered.

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

[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 compounds, lipid vesicles, micelles, millispheres, 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 a salt thereof, or the above composition, or the above delivery system or sustained-release system.

[0048] In some embodiments, the dosage form of the cosmetic comprises ointment, 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 a use of the peptide represented by the above formula (I), or its stereoisomer, or a mixture of its stereoisomers, or a salt thereof, or the above composition, or the above delivery system or sustained-release system in the preparation of a composition for caring for or treating the skin.

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

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

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

[0053] In the present disclosure, the term "skin" is understood to refer to the multiple layers that make it up, from the uppermost layer or stratum corneum to the lowermost layer or subcutaneous tissue, both ends are included. 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 the maintenance and care of the skin, improving the condition of the skin, making it delicate, smooth, tender and healthy.

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

[0056] The peptide disclosed in the present invention is obtained through artificial design, is easy to synthesize, and has high safety. It can reduce the content of acetyl-CoA carboxylase, inhibit the synthesis or secretion of skin oil, has the effects of oil control, acne removal, and repair, and can be used to care for or treat the skin and improve skin problems. BRIEF DESCRIPTION OF THE DRAWINGS

[0057] In order to more clearly illustrate the technical solution of the present disclosure, the drawings required for use in 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 ordinary technicians in this field, other drawings can be obtained based on these drawings without paying any creative work.

[0058] Figure 1 Peptide (5) Pal-Val-Ser-Gly-Leu-Thr-Pro-NH 2 Mass spectrum of .

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

[0060] Figure 3 This is a graph showing the effect of the test samples on the oil secretion of sebaceous gland cells.

[0061] Figure 4 This is a graph showing the effect of the test samples on the content of acetyl-CoA carboxylase. DETAILED DESCRIPTION

[0062] In order to make the objects, features and advantages of the present disclosure more obvious and understandable, the present disclosure is further described in detail below in conjunction with the accompanying drawings and embodiments. Obviously, the described embodiments are part of the embodiments of the present disclosure, rather than all of the embodiments. Based on the embodiments in the present disclosure, all other embodiments obtained by ordinary technicians in this field without creative work are within the scope of protection of the claims attached to the present disclosure.

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

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

[0065] Amide Resin: a starting resin for peptide synthesis (cross-linking degree 1%, substitution degree 1.72mmol / g, particle size 100-200 mesh); 2-CTC Resin: a starting resin for peptide synthesis (2-chlorotrityl chloride resin); Fmoc-Linker: 4-[(2,4-dimethoxyphenyl)(Fmoc-amino)methyl]phenoxyacetic acid; Ac 2 O: 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] Example 1 Pal-Val-Ser-Gly-Leu-Thr-Pro-NH 2 Preparation

[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 remove the solvent.

[0069] Weigh 25g of Fmoc-Linker and 7.5g of HOBt into a dry Erlenmeyer flask, dissolve them in DMF solvent, and prefreeze them at -20℃ for 10min, then add 10.7mL of DIC to activate them for 10min.

[0070] The activated Fmoc-Linker was added to the swollen resin and reacted for 3 h. The reaction solution was removed, the resin was washed, and the solvent was removed.

[0071] Continue to join Ac 2 O and DIPEA capping for 2 h. Wash the resin and remove the solvent.

[0072] 1.2 Removal of Fmoc

[0073] Fmoc-Linker-Amide Resin was de-Fmoced twice with 20% piperidine / DMF, each time for 10 min, and samples were taken for K testing, showing dark blue color. The resin was washed with DMF for 7 times, and the solvent was removed.

[0074] 1.3 Feeding reaction

[0075] Weigh 20.3g of Fmoc-Pro-OH and 9.7g of HOBt into a dry conical flask, add DMF to dissolve it, seal it and place it in a -18℃ refrigerator for 30min. Add 13.9mL of DIC to activate for 10min, avoiding moisture. Add the activated amino acid to the deprotected resin and react for 1h, then remove the reaction solution. The K test resin is colorless and transparent, indicating that the reaction is complete.

[0076] The N-terminal Fmoc group was deprotected, and 23.9 g of Fmoc-Thr(tBu)-OH after activation was coupled to the peptidyl resin using DMF as solvent in the presence of 9.7 g HOBt and 13.9 mL DIC, and the reaction was continued for 1 h. The resins were then washed and the deprotection of the Fmoc group was repeated to couple the next amino acid. In each coupling, 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 were sequentially coupled in the presence of 9.7 g HOBt and 13.9 mL DIC using DMF as solvent; after the reaction was complete, the resin was washed and the solvent was removed.

[0077] The N-terminal Fmoc group of the peptidyl resin was deprotected by using 20% ​​piperidine / DMF for two times, each time for 10 min, and the sample was taken for K test, and the color was dark blue. The resin was washed with DMF for 6 times, and the solvent was removed.

[0078] In the presence of 16.2 g HOBt and 23.1 mL DIC, 25.7 g of Pal-OH was coupled to the peptidyl resin using DMF as solvent. The reaction was continued for 1 h. The resin was washed, the solvent was removed, and 52.5 g of Pal-Val-Ser(tBu)-Gly-Leu-Thr(tBu)-Pro-Linker-Amide Resin was obtained after shrinkage and drying.

[0079] 1.4 Lysis

[0080] 114 mL of TFA, 3 mL of TIS and 3 mL of water were measured, mixed and stirred evenly to obtain a cleavage solution, which was sealed and placed in a -20°C refrigerator for later use; isopropyl ether was placed in a -20°C refrigerator for later use.

[0081] Weigh 20g of Pal-Val-Ser(tBu)-Gly-Leu-Thr(tBu)-Pro-Linker-Amide Resin, add it to a round-bottom flask, add the frozen lysate, and stir for 2h. Filter and collect the filtrate, concentrate it to 60mL, add 900mL of isopropyl ether to precipitate, stir and centrifuge with isopropyl ether for 6 times, and dry it in vacuum to obtain 7.5g of Pal-Val-Ser-Gly-Leu-Thr-Pro-NH 2 Crude peptide.

[0082] 1.5 Purification

[0083] Weigh 7.5 g of Pal-Val-Ser-Gly-Leu-Thr-Pro-NH 2 The crude peptide was dissolved in acetic acid: methanol (V: V = 6: 1) solution, filtered with a microporous filter membrane with a pore size of 0.22 μm, and then 10 mL of pure water was added to precipitate the gel. The gel was heated to 50°C to obtain a clear and transparent solution. The sample was loaded and purified by reverse phase HPLC. The purification gradient is as follows:

[0084]

[0085]

[0086] The sample was purified, the fractions were collected, concentrated and lyophilized to obtain a peptide (5) with a purity greater than 95% Pal-Val-Ser-Gly-Leu-Thr-Pro-NH 2 .

[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 remove the solvent.

[0090] 2.2 Feeding reaction

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

[0092] Fmoc-Pro-2-CTC Resin was de-Fmoced twice with 20% piperidine / DMF, each time for 10 min, and samples were taken for K testing, showing dark blue color. The resin was washed with DMF for 7 times, and the solvent was removed.

[0093] Weigh 14.4g of Fmoc-Thr(tBu)-OH and 5.9g of HOBt into a dry conical flask, add DMF to dissolve it, seal it and place it in a -18℃ refrigerator for 30min. Add 8.4mL of DIC to activate it for 10min. Add the activated amino acid to the deprotected resin and react for 1.5h, then remove the reaction liquid. The K test resin is colorless and transparent, indicating that the reaction is complete.

[0094] The N-terminal Fmoc group was deprotected, and 12.8 g of Fmoc-Leu-OH after activation was coupled to the peptidyl resin using DMF as solvent in the presence of 5.9 g HOBt and 8.4 mL DIC, and the reaction was continued for 1.5 h. The resins were then washed and the deprotection of the Fmoc group was repeated to couple the next amino acid. In each coupling, 12.6 g of Fmoc-Gly-OH, 16.1 g of Fmoc-Ser(tBu)-OH, and then 14.3 g of Fmoc-Val-OH were sequentially coupled in the presence of 6.9 g HOBt and 9.8 mL DIC using DMF as solvent; after the reaction was complete, the resin was washed and the solvent was removed.

[0095] The N-terminal Fmoc group of the peptidyl resin was deprotected by using 20% ​​piperidine / DMF for two times, each time for 10 min, and the sample was taken for K test, and the color was dark blue. The resin was washed with DMF for 7 times, and the solvent was removed.

[0096] In the presence of 9.8 g HOBt and 13.9 mL DIC, 15.5 g of Pal-OH was coupled to the peptidyl resin using DMF as solvent. The reaction was continued for 1 h. The resin was washed, the solvent was removed, and 39.5 g of Pal-Val-Ser(tBu)-Gly-Leu-Thr(tBu)-Pro-2-CTC Resin was obtained after shrinkage and drying.

[0097] 2.3 Lysis

[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 39.5g of Pal-Val-Ser(tBu)-Gly-Leu-Thr(tBu)-Pro-2-CTC Resin, add it to a round-bottom flask, add the frozen lysate, and stir to react for 2h. Filter by suction, collect the filtrate and concentrate it to 120mL, add 1.2L of pure water to precipitate, filter the white solid with a Buchner funnel, and wash it twice with pure water to obtain 13g of Pal-Val-Ser-Gly-Leu-Thr-Pro-OH crude peptide.

[0100] 2.4 Purification

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

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

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

[0104] The test results of peptide (6) are shown in Figure 2 , the results show that [M+H] + The mass-to-charge ratio (m / z) of the 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 culture medium, fetal bovine serum, phosphate buffered saline (PBS), palmitic acid and linoleic acid mixed solution (FFA), Oil Red O staining kit.

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

[0109] (1) Prepare 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 allow to solidify at room temperature after it is completely dissolved. Then add 0.5 mL of isopropanol and continue heating until it dissolves again. Finally, add 1 mL of PBS to make a 2 mL system for later use.

[0110] (2) Prepare 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.

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

[0112] 3.2 Instruments

[0113] ELISA reader, CO 2 Incubator, clean bench.

[0114] 3.3 Cell lines

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

[0116] 3.4 Samples to be tested

[0117] Experimental groups: peptide (5) and peptide (6), tested at concentrations of 5 ppm, 10 ppm, 25 ppm, and 50 ppm.

[0118] Control group: PBS.

[0119] Model Group:FFA.

[0120] 3.5 Experimental methods

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

[0122] Take an appropriate amount of cell suspension and inoculate it into a 12-well plate and place it in a constant temperature CO 2 Culture in an incubator for 24 h.

[0123] Except for the control group, which was added with PBS, FFA was added to each well to induce modeling. Then, different concentrations of the test samples were added to the experimental groups, and the cells were kept at a constant temperature of CO. 2 Culture in an incubator for 48 h.

[0124] The culture medium was discarded and the relative content of cellular lipid secretion was calculated according to the instructions of the Oil Red O staining kit.

[0125] 3.6 Experimental Results

[0126] Oil Red O is a fat-soluble dye that is highly soluble in fat. The principle of its staining is that Oil Red O can specifically adsorb neutral triglycerides, lipids, and lipoproteins in tissues and cells to stain fat. In this experiment, the test sample was used to treat the cells induced by FFA, and the oil secretion of the corresponding cells was detected to determine whether the peptide disclosed in the present invention can inhibit oil secretion.

[0127] The results of the test samples' effects on sebaceous gland cell oil secretion are shown in Figure 3 . The results showed that compared with the control group, the oil secretion of the model group increased significantly, indicating that the modeling was successful. Compared with the model group, the experimental group was able to significantly inhibit the oil secretion of sebaceous gland cells. It can be seen that the peptide disclosed in the present invention can inhibit the oil secretion of the skin, slow down the oil deposition, reduce the occurrence of acne or acne, and also help the recovery of the skin after acne or acne occurs, and has the effects of oil control, acne removal and repair.

[0128] Example 4 Acetyl-CoA Carboxylase Content Test

[0129] 4.1 Reagents and Materials

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

[0131] 4.2 Instruments

[0132] ELISA reader, CO 2 Incubator, clean bench, vortexer.

[0133] 4.3 Cell lines

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

[0135] 4.4 Samples to be tested

[0136] Experimental groups: peptide (5) and peptide (6), tested at concentrations of 5 ppm and 10 ppm.

[0137] Control group: PBS.

[0138] Model Group:FFA.

[0139] 4.5 Experimental methods

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

[0141] The cell suspension was inoculated on a 12-well plate and placed in a constant temperature CO 2 Culture in an incubator for 24 h.

[0142] Except for the control group, which was added with PBS, FFA was added to each well to induce modeling. Then, different concentrations of the test samples were added to the experimental groups, and the cells were kept at a constant temperature of CO. 2 Culture in an incubator for 48 h.

[0143] Collect cells, centrifuge to obtain cell pellets, add RIPA lysis buffer, shake with a vortexer 3 times (30s / time, 3min interval), centrifuge at 12000rpm*10min. Pipette the supernatant, operate according to the ELISA operating instructions, and use the BCA protein quantification kit to detect the total protein concentration of the supernatant and calculate the acetyl-CoA carboxylase content.

[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 sebum reserves in sebaceous glands, and thus alleviate seborrhea. In this experiment, the test sample was used to treat the cells induced by FFA, and the content of acetyl-CoA carboxylase in the corresponding cells was detected to determine whether the peptide disclosed in the present invention can inhibit the expression of acetyl-CoA carboxylase.

[0146] The results of the test samples on the content of acetyl-CoA carboxylase are shown in Figure 4 The results showed that compared with the control group, the content of acetyl-CoA carboxylase in the model group was significantly increased. Compared with the model group, the experimental groups were able to significantly reduce the content of acetyl-CoA carboxylase, indicating that the peptide disclosed in the present invention can inhibit the expression of acetyl-CoA carboxylase, thereby inhibiting the synthesis of fats and oils.

[0147] It can be seen that the peptide disclosed in the present invention can reduce the content of acetyl-CoA carboxylase, inhibit the synthesis or secretion of skin oil, slow down oil deposition, reduce the occurrence of acne or acne, and also help the recovery of the skin after acne or acne occurs, and has the effects of oil control, acne removal and repair.

[0148] Example 5 Emulsion containing peptide (5)

[0149]

[0150]

[0151] Preparation method: According to the formula and dosage, heat the materials of phase A to 75-80℃ in the oil phase pot, and stir to dissolve completely; add the materials of phase B into the emulsifying pot, stir and heat to 80-85℃, and dissolve completely; 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℃, add materials of phase C and phase D and continue stirring; cool to 35-40℃, add materials of phase E, and stir for 10-15 minutes.

[0152] Example 6 Essence containing peptide (6)

[0153]

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

[0155] Example 7 Cream containing peptide (1)

[0156]

[0157]

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

[0159] In the present disclosure, relational terms such as first and second, etc. are used only to distinguish one entity or operation from another entity or operation, and do not necessarily require or imply any such actual relationship or order between these entities or operations. Moreover, the terms "include", "comprise" or any other variants thereof are intended to cover non-exclusive inclusion, so that a process, method, article or terminal device including a series of elements includes not only those elements, but also other elements not explicitly listed, or also includes elements inherent to such process, method, article or terminal device. In the absence of further restrictions, the elements defined by the statement "comprise one..." do not exclude the presence of other identical elements in the process, method, article or terminal device including the elements.

[0160] Although the specific embodiments of the present disclosure are described for the purpose of illustration, various modifications or improvements may be made by those skilled in the art without departing from the spirit and scope of the present disclosure. These modifications or improvements should all fall within the scope of the appended claims of the present disclosure.

Claims

1. A 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 H, 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, characterized in that: Selected from the following peptides (1)-(8): (1)H-Val-Ser-Gly-Leu-Thr-Pro-NH2; (2)H-Val-Ser-Gly-Leu-Thr-Pro-OH; (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), wherein 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, wherein the organic base includes: ethylamine, diethylamine, ethylenediamine, ethanolamine, diethanolamine, 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, wherein 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; and the inorganic acid includes hydrochloric acid, sulfuric acid, boric acid or carbonic acid.

4. A composition, characterized in that The composition comprises an effective amount of a peptide represented by formula (I) or a salt thereof according to any one of claims 1 to 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 a peptide represented by formula (I) or a salt thereof according to any one of claims 1 to 3, or a 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, millicapsules, microcapsules, nanocapsules, nanostructured lipid carriers, sponges, inclusion compounds, lipid vesicles, micelles, millispheres, microspheres, nanospheres, lipid spheres, microemulsions, nanoemulsions, milliparticles, microparticles or nanoparticles.

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

8. The cosmetic according to claim 7, characterized in that: The dosage forms of the cosmetics include ointments, creams, emulsions, aqueous solutions, oils, gels, powders, tablets, muds, patches, films, aerosols, sprays, freeze-dried preparations or nano preparations.

9. Use of the peptide represented by formula (I) or a salt thereof according to any one of claims 1 to 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 composition for caring for or treating skin.

10. Use of the peptide represented by formula (I) or a salt thereof according to any one of claims 1 to 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 composition for oil control, acne removal or repair.

11. Use of the peptide represented by formula (I) or a salt thereof according to any one of claims 1 to 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 composition for inhibiting skin oil synthesis or secretion, or in the preparation of a composition for reducing the content of acetyl-CoA carboxylase.

12. Use of the peptide represented by formula (I) or a salt thereof according to any one of claims 1 to 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 cosmetics.

Images