Sebum-regulating polypeptide, and composition thereof and use thereof

Abstract

The present invention relates to the technical field of polypeptides. Disclosed are a sebum-regulating polypeptide, and a composition thereof and the use thereof. The peptide or a salt thereof has the structure as represented by formula (I): R1-Val-Ser-Gly-Leu-Thr-Pro-R2(I). The peptide or a stereoisomer thereof, or a mixture of the stereoisomer thereof, or a salt thereof, or a composition thereof has sebum-regulating, anti-acne, and skin-repairing effects.

Description

An oil-controlling polypeptide, its composition and uses

[0001] This application is based on and claims priority to CN application No. 202411851184.2, filed on December 16, 2024, the disclosure of which is incorporated herein by reference in its entirety. Technical Field

[0002] This disclosure relates to the field of polypeptide technology, and in particular to an oil-controlling polypeptide and compositions thereof, and their uses. Background Technology

[0003] Sebaceous glands are multi-acini holoseric secretory tissues composed of one or more sac-like acini and short ducts, distributed throughout 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 substance (sebum) onto 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 radiation, or endocrine disorders, can cause sebaceous gland hyperfunction, leading to excessive sebum secretion. This results in oily, shiny hair and skin, and may even cause clogged pores, blackheads or whiteheads, acne, and other skin problems.

[0004] Human sebum is mainly composed of triglycerides, wax esters, free fatty acids, squalene, and cholesterol. The formation of triglycerides, wax esters, and free fatty acids is closely related to fatty acid synthesis. Acetyl-CoA carboxylase (ACC) is a biotinylate isolated from the enzyme system that synthesizes long-chain fatty acids from acetyl-CoA and plays a crucial role in fatty acid synthesis. In vivo, acetyl-CoA carboxylase uses energy provided by ATP to catalyze the conversion of acetyl-CoA to malonyl-CoA. Then, malonyl-CoA acts as a two-carbon unit donor, synthesizing fatty acids under the action of fatty acid carbon chain elongation enzymes. Therefore, inhibiting the activity or expression of acetyl-CoA carboxylase can limit fatty acid synthesis, inhibit lipid production in sebaceous cells, thereby alleviating sebum secretion and controlling oil production.

[0005] Currently, there are relatively few peptides with oil-controlling effects, and it is necessary to study more oil-controlling peptide compounds. Summary of the Invention

[0006] This disclosure relates to a peptide, and compositions containing such peptides, which have skin care or therapeutic effects.

[0007] On the one hand, this disclosure provides a peptide of formula (I), or a stereoisomer thereof, or a mixture thereof, or a salt thereof.

[0008] R1-Val-Ser-Gly-Leu-Thr-Pro-R2(I)

[0009] In formula (I),

[0010] R1 is selected from H or R3-CO-, where R3 is selected from substituted or unsubstituted alkyl or substituted or unsubstituted alkenyl groups;

[0011] R2 is selected from: -NR4R5 or -OR4, wherein each R4 and R5 is independently selected from: H, substituted or unsubstituted alkyl, substituted or unsubstituted alkenyl;

[0012] The alkyl group refers to a saturated aliphatic straight-chain or branched alkyl group having 1-24 carbon atoms (or 1-16 carbon atoms; or 1-14 carbon atoms; or 1-12 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;

[0013] The alkenyl group refers to a straight-chain 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 by a single bond; in some embodiments, the alkenyl group is selected from: vinyl, oleyl, or linoleyl;

[0014] In some embodiments, the substituents in "substituted alkyl" and "substituted alkenyl" are selected from C1-C4 alkyl; hydroxyl; C1-C4 alkoxy; amino; C1-C4 aminoalkyl; C1-C4 carbonyloxy; C1-C4 oxycarbonyl; halogens (such as fluorine, chlorine, bromine, and iodine); cyano; nitro; azide; C1-C4 alkylsulfonyl; thiol; C1-C4 alkylthio; C6-C 30 Aryl groups, such as phenoxy groups; -NR b (C=NR b )NR b R c , where R b and R c It is independently selected from: H, C1-C4 alkyl, C2-C4 alkenyl, C2-C4 alkynyl, C3-C 10 cycloalkyl, C6-C 18Aryl, C7-C 17 Aryl groups, or protecting groups with three to ten-membered heterocyclic groups or amino groups.

[0015] 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.

[0016] 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.

[0017] In some embodiments, R1 is H, acetyl, lauroyl, myristoyl, or palmitoyl; R2 is -OH or -NH2.

[0018] In some embodiments, the peptide represented by formula (I), or a stereoisomer thereof, a mixture thereof, or a salt thereof, is selected from peptides (1)-(8):

[0019] (1) H-Val-Ser-Gly-Leu-Thr-Pro-NH2;

[0020] (2)H-Val-Ser-Gly-Leu-Thr-Pro-OH;

[0021] (3) Ac-Val-Ser-Gly-Leu-Thr-Pro-NH2;

[0022] (4) Ac-Val-Ser-Gly-Leu-Thr-Pro-OH;

[0023] (5)Pal-Val-Ser-Gly-Leu-Thr-Pro-NH2;

[0024] (6)Pal-Val-Ser-Gly-Leu-Thr-Pro-OH;

[0025] (7) Myr-Val-Ser-Gly-Leu-Thr-Pro-NH2;

[0026] (8) Myr-Val-Ser-Gly-Leu-Thr-Pro-OH.

[0027] The peptide represented by formula (I) of this disclosure can exist as a stereoisomer or a mixture of 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 diastereomeric or enantiomers, depending on the number of asymmetric carbons and the presence of any isomers or isomeric mixtures. In some embodiments, the peptide represented by formula (I) of this disclosure has a structure that is a pure isomer, i.e., an enantiomer or diastereomeric isomer. In some embodiments, the peptide represented by formula (I) of this disclosure has a structure that is an L-isomer.

[0028] This disclosure also includes all suitable isotopic variants of the peptide represented by formula (I). These isotopic variants of the peptides of this disclosure are understood herein to refer to compounds in which at least one atom within the peptide of this disclosure is replaced by another atom of the same atomic number, but said other atom has an atomic mass different from that of atoms commonly or predominantly found in nature. Examples of isotopes that can be incorporated into the peptides of this disclosure are those of hydrogen, carbon, nitrogen, or oxygen, for example… 2 H (deuterium) 3 H (tritium) 13 C 14 C 15 N、 17 O or 18 O. Specific isotopic variants of the peptides disclosed herein (especially those that have been incorporated with one or more radioactive isotopes) may be advantageous, for example, in examining mechanisms of action or the distribution of active compounds in vivo; due to their relatively simple prepareability and detectability, especially with 3 H or 14 Compounds labeled with the C isotope are suitable for this purpose. Additionally, the incorporation of isotopes (e.g., deuterium) can produce specific therapeutic benefits, such as prolonged in vivo half-life or reduced required active dose, due to the enhanced metabolic stability of the compounds. Isotopic variants of the peptides disclosed herein can be prepared by methods known to those skilled in the art, such as those further described below and those described in the examples, using respective reagents and / or corresponding isotope modifiers of the starting materials.

[0029] The term "salt" refers to a salt recognized for use in animals, and more precisely in humans, including metal salts of peptides of formula (I), wherein the metal includes, but is not limited to: lithium, sodium, potassium, calcium, magnesium, manganese, copper, zinc, or aluminum; salts formed by peptides of formula (I) with organic bases, wherein the organic bases include, but are not limited to: ethylamine, diethylamine, ethylenediamine, ethanolamine, diethanolamine, arginine, lysine, histidine, or piperazine; salts formed by peptides of formula (I) with inorganic or organic acids, wherein 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; and wherein the inorganic acids include, but are not limited to: hydrochloric acid, sulfuric acid, boric acid, or carbonic acid.

[0030] The synthesis of the peptide or salt thereof shown in this disclosure (I) 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 and liquid methods, or by biotechnological methods aimed at producing a desired sequence, or by controlled hydrolysis of proteins of animal, fungal or plant origin.

[0031] For example, a method for obtaining the peptide represented by formula (I) includes the following steps:

[0032] - Couple amino acids with a protected N-terminus and a free C-terminus to amino acids with a free N-terminus and a protected C-terminus or a C-terminus bound to a solid support.

[0033] - Eliminate the groups protecting the N-terminus;

[0034] - Repeat this coupling sequence and remove the group protecting the N-terminus until the desired peptide sequence is obtained;

[0035] - Eliminate the groups protecting the C-terminus or cleave them from the solid support.

[0036] 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 to an amino acid having a free N-terminus and a C-terminus bound to a polymer support; removing the group protecting the N-terminus; and repeating this sequence a number of times as required to thus obtain a peptide of the desired length, followed by cleaving the synthesized peptide from the original polymer support.

[0037] Throughout the synthesis, the functional groups of the side chains of these amino acids are adequately protected with temporary or permanent protecting groups and can be deprotected simultaneously or orthogonally with the process of cleaving the peptide from the polymer carrier.

[0038] In some embodiments, solid-phase synthesis can be carried out using a convergent strategy, which involves coupling dipeptides or tripeptides to a polymer support or to dipeptides or amino acids previously bound to a polymer support.

[0039] The N- and C-termini are deprotected and / or cleaved from the polymer support in an indeterminate order using standard conditions and methods known in the art, after which the functional groups at the ends can be modified. Optional N- and C-terminal modifications can be made to the peptide bound to the polymer support, or to the peptide after cleavage from the polymer support.

[0040] In another aspect of this disclosure, a composition is provided comprising an effective amount of a peptide represented by the above formula (I), or a stereoisomer thereof, or a mixture thereof, or a salt thereof, and at least one excipient and optionally an adjuvant.

[0041] In some embodiments, the adjuvant includes, but is not limited to: agents that inhibit acetyl-CoA carboxylase activity, analgesics, agents that inhibit PAR-2 ​​activity, collagen synthesis stimulants, agents that regulate PGC-1α synthesis, agents that regulate PPARγ activity, agents that increase or decrease triglyceride content in adipocytes, agents that stimulate or delay adipocyte differentiation, lipolytic agents or agents that stimulate lipolysis, lipogenic agents, inhibitors of acetylcholine receptor aggregation, agents that inhibit muscle contraction, anticholinergic agents, elastase inhibitors, matrix metalloproteinase inhibitors, melanin synthesis stimulants or inhibitors, whitening agents or depigmenting agents, pigmentation promoters, self-tanning agents, NO-synthesizers, and 5α-reductase inhibitors. Inhibitors of proenzymes, inhibitors of lysyl hydroxylase and / or prolyl hydroxylase, antioxidants, anti-air pollution agents, anti-glycation agents, antihistamines, antiparasitic agents, emollients, organic solvents, liquid propellants, water-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 that reduce or treat under-eye bags, keratolytic agents, antimicrobial agents, agents that stimulate elastin synthesis, agents that stimulate core proteoglycan synthesis, agents that stimulate laminin synthesis, agents that stimulate defensin synthesis, agents that stimulate chaperone protein synthesis, and cAMP stimulants. Synthetic agents, agents stimulating hyaluronic acid synthesis, agents stimulating fibronectin synthesis, agents stimulating deacetylase synthesis, agents stimulating lipid and stratum corneum component synthesis, ceramides, fatty acids, agents inhibiting collagen degradation, agents inhibiting elastin degradation, agents inhibiting serine proteases, agents stimulating fibroblast proliferation, agents stimulating keratinocyte proliferation, agents stimulating adipocyte proliferation, agents stimulating melanocyte proliferation, agents stimulating keratinocyte differentiation, agents inhibiting acetylcholinesterase, skin relaxants, agents stimulating glycosaminoglycan synthesis, anti-hyperkeratosis agents, comedolytic agents, anti-psoriasis agents, anti-eczema agents, DNA repair agents, DNA protectants, stabilizers, antipruritics, and so on. Coagulants, firming agents, reconstructing agents, agents that regulate sebum production, antiperspirants, agents that stimulate healing, agents that assist healing, agents that stimulate re-epithelialization, agents that assist re-epithelialization, cytokines, sedatives, anti-inflammatory agents, anesthetics, agents that act on capillary circulation and / or microcirculation, agents that stimulate angiogenesis, agents that inhibit vascular permeability, venous tension agents, agents that act on cell metabolism, agents used to improve dermal-epidermal junction, agents that induce hair growth, agents that inhibit or delay hair growth, fragrances, chelating agents, plant extracts, essential oils, marine extracts, agents derived from bio-fermentation processes, inorganic salts, cell extracts, and organic or inorganic photoprotective agents or mixtures thereof that are effective against UVA and / or UVB.

[0042] The effective amount of the disclosed peptides to be administered and their dosage will depend on many factors, including age, the user's condition, the severity of the condition, the route and frequency of administration, and the specific nature of the peptide to be used.

[0043] "Effective amount" means an amount of one or more peptides of this disclosure that is non-toxic but sufficient to provide the desired effect. The peptides of this disclosure are used in compositions of this 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.

[0044] Another aspect of this disclosure provides a delivery system or sustained-release system for better penetration of an active ingredient, comprising an effective amount of the peptide represented by formula (I) above, or a stereoisomer thereof, a mixture of stereoisomers thereof, a salt thereof, or a combination thereof.

[0045] The term "delivery system" refers to a diluent, adjuvant, excipient, or carrier applied with the peptides of this disclosure, selected from water, oil, or surfactants, including those of petroleum, animal, plant, or synthetic origin, such as and not limited to peanut oil, soybean oil, mineral oil, sesame oil, castor oil, polysorbate, sorbitol ester, ether sulfate, sulfate, betaine, glucosinolate, maltodextrin, fatty alcohol, nonyl alcohol ether, poloxamer, polyoxyethylene, polyethylene glycol, dextran, glycerol, digitalis saponins, and the like. Those skilled in the art are familiar with various diluents, adjuvants, excipients, or carriers that can be used in different delivery systems for administering the peptides of this disclosure.

[0046] The term "sustained release" is used in its conventional sense to refer to a compound delivery system that provides the gradual release of a compound over a period of time. In some embodiments, the sustained release system maintains a relatively constant level of compound release throughout the entire time period.

[0047] Examples of delivery systems or sustained-release systems include, but are not limited to: liposomes, oil bodies, alcohol bodies, millimeter capsules, micrometer capsules, nanocapsules, nanostructured lipid carriers, sponges, inclusion complexes, lipid vesicles, micelles, millimeter spheres, micrometer spheres, nanospheres, lipid spheres, micrometer emulsions, nanoemulsions, millimeter particles, micrometer particles, or nanoparticles.

[0048] In another aspect of this disclosure, a cosmetic product is provided, comprising an effective amount of the peptide represented by the above formula (I), or a stereoisomer thereof, a mixture of stereoisomers thereof, a salt thereof, or the above-described composition, or the above-described delivery system or sustained-release system.

[0049] In some embodiments, the dosage form of the cosmetic includes ointment, cream, emulsion, aqueous solution, oil, gel, powder, tablet, mud, patch, film, aerosol, spray, lyophilized preparation or nano-preparation.

[0050] Another aspect of this disclosure provides the use of a peptide of formula (I) above, or a stereoisomer thereof, or a mixture of stereoisomers thereof, or a salt thereof, or a composition thereof, or a delivery system or sustained-release system thereof, in the preparation of a composition for the care or treatment of skin.

[0051] Another aspect of this disclosure provides the use of a peptide of formula (I) above, or a stereoisomer thereof, or a mixture of stereoisomers thereof, or a salt thereof, or the above-described composition, or the above-described delivery system or sustained-release system, in the preparation of a composition for oil control, acne treatment or repair.

[0052] In another aspect of this disclosure, there is a use of the peptide represented by the above formula (I), or a stereoisomer thereof, or a mixture of stereoisomers thereof, or a salt thereof, or the above-described composition, or the above-described delivery system or sustained-release system, in the preparation of a composition for inhibiting the synthesis or secretion of skin sebum, or in the preparation of a composition for reducing the content of acetyl-CoA carboxylase.

[0053] Another aspect of this disclosure provides the use of a peptide of formula (I) above, or a stereoisomer thereof, or a mixture of stereoisomers thereof, or a salt thereof, or the above-described composition, or the above-described delivery system or sustained-release system in the preparation of cosmetics.

[0054] In this disclosure, the term "skin" should be understood as comprising its multiple layers, from the uppermost layer or stratum corneum to the lowermost layer or subcutaneous tissue, including both ends. These layers are composed of different types of cells, such as keratinocytes, fibroblasts, melanocytes, and / or adipocytes. In this disclosure, the term "skin" includes the scalp.

[0055] The term "skin care" refers to the maintenance and care of the skin to improve its condition, making it delicate, smooth, soft, and healthy.

[0056] This disclosure has the following advantages and effects:

[0057] The peptide disclosed herein is artificially designed, easy to synthesize, and highly safe. It can reduce the content of acetyl-CoA carboxylase and inhibit the synthesis or secretion of skin oil, thus having the effects of oil control, acne removal, and repair. It can be used to care for or treat the skin and improve skin problems. Attached Figure Description

[0058] To more clearly illustrate the technical solutions of this disclosure, the accompanying drawings used in the description of this disclosure will be briefly introduced below. Obviously, the accompanying drawings described below are only some embodiments of this disclosure. For those skilled in the art, other drawings can be obtained based on these drawings without creative effort.

[0059] Figure 1 is the mass spectrum of peptide (5) Pal-Val-Ser-Gly-Leu-Thr-Pro-NH2.

[0060] Figure 2 is the mass spectrum of peptide (6) Pal-Val-Ser-Gly-Leu-Thr-Pro-OH.

[0061] Figure 3 shows the results of the test sample's effect on sebum secretion from sebaceous gland cells.

[0062] Figure 4 shows the effect of the test samples on the content of acetyl-CoA carboxylase. Detailed Implementation

[0063] To make the objects, features, and advantages of this disclosure more apparent and understandable, the disclosure will be further described in detail below with reference to the accompanying drawings and embodiments. Obviously, the described embodiments are only some, not all, of the embodiments of this disclosure. All other embodiments obtained by those skilled in the art based on the embodiments of this disclosure without inventive effort are within the scope of protection of the appended claims.

[0064] In this disclosure, the abbreviations used 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).

[0065] Unless otherwise specified, all experimental reagents and materials used in this disclosure are commercially available. The following are abbreviations for some of the reagents and materials:

[0066] Amide Resin: A starting resin for peptide synthesis (1% crosslinking, 1.72 mmol / g substitution, 100-200 mesh particle size); 2-CTC Resin: A starting resin for polypeptide synthesis (2-chlorotriphenylmethyl 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.

[0067] Example 1: Preparation of Pal-Val-Ser-Gly-Leu-Thr-Pro-NH2

[0068] 1.1 Preparation of Fmoc-Linker-Amide Resin

[0069] Weigh 20g of Amide Resin into a solid-phase synthesis reaction column, pour in 200mL of DCM to swell for 30min, wash the resin, and remove the solvent.

[0070] Weigh 25g of Fmoc-Linker and 7.5g of HOBt into a dry Erlenmeyer flask. Dissolve in DMF solvent and pre-freeze at -20℃ for 10min. Activate with 10.7mL of DIC for 10min.

[0071] The activated Fmoc-Linker was added to the swollen resin and reacted for 3 hours. The reaction solution was then removed, the resin was washed, and the solvent was removed.

[0072] Continue adding Ac2O and DIPEA for end-capping treatment for 2 hours. Wash the resin and remove the solvent.

[0073] 1.2 De-Fmoc

[0074] Fmoc-Linker-Amide Resin was used to remove Fmoc twice with 20% piperidine / DMF, 10 min each time. Samples were taken for K-test, and the color development was deep blue. The resin was then washed 7 times with DMF, and the solvent was removed.

[0075] 1.3 Feeding and Reaction

[0076] Weigh 20.3 g of Fmoc-Pro-OH and 9.7 g of HOBt into a dry Erlenmeyer flask, add DMF to dissolve them, seal and place in a -18°C freezer for 30 min. Add 13.9 mL of DIC to activate for 10 min, avoiding moisture. Add the activated amino acids to the deprotected resin and react for 1 h, then remove the reaction solution. A colorless and transparent K test indicates complete reaction.

[0077] The N-terminal Fmoc group was deprotected, and 23.9 g of activated Fmoc-Thr(tBu)-OH was coupled to a peptide resin in the presence of 9.7 g HOBt and 13.9 mL DIC, using DMF as a solvent, and the reaction was continued for 1 h. The resin was then washed, and the deprotection treatment 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 subsequently 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 a solvent; after the reaction was complete, the resin was washed, and the solvent was removed.

[0078] To deprotect the N-terminal Fmoc group of the peptide resin, the Fmoc was removed twice with 20% piperidine / DMF, 10 min each time. A sample was taken for K testing; the result was a deep blue color. The resin was then washed six times with DMF, and the solvent was removed.

[0079] In the presence of 16.2 g HOBt and 23.1 mL DIC, 25.7 g of Pal-OH was coupled to a peptide resin using DMF as a solvent. The reaction was continued for 1 h. The resin was washed, the solvent was removed, and the resin was dried to obtain 52.5 g of Pal-Val-Ser(tBu)-Gly-Leu-Thr(tBu)-Pro-Linker-Amide Resin.

[0080] 1.4 Pyrolysis

[0081] Measure 114 mL of TFA, 3 mL of TIS and 3 mL of water, mix and stir well to obtain the lysis solution, seal and store at -20℃ for later use; store isopropyl ether at -20℃ for later use.

[0082] Weigh 20g of Pal-Val-Ser(tBu)-Gly-Leu-Thr(tBu)-Pro-Linker-Amide Resin and add it to a round-bottom flask. Add the frozen lysis buffer and stir for 2 hours. Filter the mixture, collect the filtrate, concentrate it to 60mL, add 900mL of isopropyl ether to settle it, wash it 6 times with isopropyl ether by stirring and centrifugation, and dry it under vacuum to obtain 7.5g of crude Pal-Val-Ser-Gly-Leu-Thr-Pro-NH2 peptide.

[0083] 1.5 Purification

[0084] 7.5 g of crude Pal-Val-Ser-Gly-Leu-Thr-Pro-NH2 peptide was weighed and dissolved in an acetic acid:methanol (V:V=6:1) solution. After filtration through a 0.22 μm microporous membrane, 10 mL of pure water was added, and a gel was precipitated. The solution was heated to 50 °C to obtain a clear and transparent solution. The solution was loaded onto the gel and purified by reversed-phase HPLC. The purification gradient is shown in the table below.

[0085] Time (min) Flow rate (mL / min) A% (acetonitrile) B% (0.1% acetic acid + pure water) 0 40 20 80 540 30 70 15 40 60 40 30 40 80 20 45 40 82 18 80 40 90 10 100 40 1000

[0086] The sample was purified, the fraction was collected, concentrated and lyophilized 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 the resin

[0089] Weigh 20g 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 and Reaction

[0091] Weigh 16.0 g of Fmoc-Pro-OH and 5.9 g of HOBt into a dry Erlenmeyer flask. Dissolve in DMF solvent and cool in an ice-water bath for 10 min. 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. Remove the reaction solution, wash the resin, and remove the solvent. Continue to add DCM, MeOH, and DIPEA for end-capping treatment for 0.5 h. Wash the resin, remove the solvent, and obtain Fmoc-Pro-2-CTC Resin.

[0092] Fmoc-Pro-2-CTC Resin was used to remove Fmoc twice with 20% piperidine / DMF, 10 min each time. Samples were taken for K-test, and the color development was deep blue. The resin was then washed 7 times with DMF, and the solvent was removed.

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

[0094] The N-terminal Fmoc group was deprotected, and 12.8 g of activated Fmoc-Leu-OH was coupled to a peptide resin in the presence of 5.9 g HOBt and 8.4 mL DIC using DMF as a solvent, with the reaction time lasting 1.5 h. The resin was then washed, and the deprotection treatment 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 subsequently 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 a solvent; after complete reaction, the resin was washed, and the solvent was removed.

[0095] To deprotect the N-terminal Fmoc group of the peptide resin, the Fmoc was removed twice with 20% piperidine / DMF, 10 min each time. A sample was taken for K testing; the color development was deep blue. The resin was washed seven times with DMF, 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 a peptide resin using DMF as a solvent. The reaction was continued for 1 h. After washing the resin, removing the solvent, and drying, 39.5 g of Pal-Val-Ser(tBu)-Gly-Leu-Thr(tBu)-Pro-2-CTC Resin was obtained.

[0097] 2.3 Pyrolysis

[0098] Measure 209 mL of TFA, 5.5 mL of TIS and 5.5 mL of water, mix and stir well to obtain the lysis solution, seal and store in a -20℃ refrigerator for later use.

[0099] Weigh 39.5 g of Pal-Val-Ser(tBu)-Gly-Leu-Thr(tBu)-Pro-2-CTC Resin and add it to a round-bottom flask. Add the frozen lysis buffer and stir for 2 h. Filter the mixture, collect the filtrate, concentrate it to 120 mL, add 1.2 L of pure water to settle it, and filter the resulting white solid using a Buchner funnel. Wash the solid twice with pure water to obtain 13 g of crude Pal-Val-Ser-Gly-Leu-Thr-Pro-OH peptide.

[0100] 2.4 Purification

[0101] Weigh 13g of crude Pal-Val-Ser-Gly-Leu-Thr-Pro-OH peptide, dissolve it in methanol, filter it, add a large amount of pure water to the filtrate, centrifuge once, add 1% dilute ammonia water and centrifuge three times, finally add pure water and wash and centrifuge once, 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 methods similar to those in Examples 1 and 2, and the molecular weight of the obtained peptides is determined by ESI-MS.

[0103] The test results for 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 for 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 was 811.5451, and the molecular weight measured by mass spectrometry was 810.55.

[0105] Example 3: Oil Content Test

[0106] 3.1 Reagents and Materials

[0107] Trypsin digestion solution, DMEM medium, fetal bovine serum, phosphate-buffered saline (PBS), palmitic acid-linolenic acid mixture (FFA), and Oil Red O staining kit.

[0108] The preparation method of the palmitic acid-linolenic acid mixed solution (FFA) is as follows:

[0109] (1) Preparation of palmitic acid solution: Weigh 3.84 mg palmitic acid, add 0.5 mL of 0.6 mg / mL NaOH solution, heat at 70°C until completely dissolved, and let it solidify at room temperature; then add 0.5 mL isopropanol and continue heating until it dissolves again; finally add 1 mL of PBS to prepare a 2 mL system for later use.

[0110] (2) Preparation of linoleic acid solution: Weigh 4.2 mg of linoleic acid, add 0.5 mL of isopropanol, 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. After dilution, take 1 mL of each solution and mix thoroughly. Filter through a 0.22 μm microporous membrane to obtain the final product.

[0112] 3.2 Instruments

[0113] Microplate reader, CO2 incubator, clean bench.

[0114] 3.3 Cell lines

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

[0116] 3.4 Sample to be tested

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

[0118] Control group: PBS.

[0119] Model group: FFA.

[0120] 3.5 Experimental Methods

[0121] Take one flask of SZ-95 cells in the exponential growth phase, add 0.25% trypsin digestion solution, digest to detach the adherent cells, and count (1-4) × 10⁻⁶ cells. 5 Cells were prepared as a suspension at a density of 1 cell per mL.

[0122] Take an appropriate amount of cell suspension and seed it into a 12-well plate, and incubate it in a constant temperature CO2 incubator for 24 hours.

[0123] Except for the control group which was given PBS, each of the other wells was given FFA to induce modeling. Then, different concentrations of the test samples were added to the experimental groups and incubated in a constant temperature CO2 incubator for 48 hours.

[0124] Discard the culture medium and follow the instructions for the Oil Red O staining kit to calculate the relative content of lipid secretion in the cells.

[0125] 3.6 Experimental Results

[0126] Oil Red O is a fat-soluble dye that is highly soluble in fat. Its staining principle is that Oil Red O specifically adsorbs onto neutral triglycerides, lipids, and lipoproteins in tissues and cells, thereby staining the fat. In this experiment, test samples were used to treat FFA-induced cells, and the amount of lipid secretion in the corresponding cells was measured to determine whether the peptide disclosed herein can inhibit lipid secretion.

[0127] The effects of the test samples on sebum secretion from sebaceous gland cells are shown in Figure 3. The results showed that, compared to the control group, the model group exhibited a significant increase in sebum secretion, indicating successful modeling. Compared to the model group, the experimental groups all showed significant inhibition of sebum secretion from sebaceous gland cells. Therefore, the peptide disclosed in this study can inhibit sebum secretion, slow down sebum deposition, reduce the occurrence of pimples or acne, and aid in skin recovery after pimple or acne develops, demonstrating oil control, acne removal, and repair effects.

[0128] Example 4: Test of Acetyl-CoA Carboxylase Content

[0129] 4.1 Reagents and Materials

[0130] Trypsin digestion solution, DMEM medium, fetal bovine serum, phosphate-buffered saline (PBS), palmitic acid-linolenic acid mixture (FFA), RIPA lysis buffer, BCA protein quantification kit, and human acetyl-CoA carboxylase 1 ELISA kit. The preparation method for FFA was the same as in Example 3.

[0131] 4.2 Instruments

[0132] Microplate reader, CO2 incubator, clean bench, vortex mixer.

[0133] 4.3 Cell lines

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

[0135] 4.4 Sample to be tested

[0136] Experimental group: peptide (5) and peptide (6), with test concentrations of 5 ppm and 10 ppm.

[0137] Control group: PBS.

[0138] Model group: FFA.

[0139] 4.5 Experimental Methods

[0140] Take one flask of cells in the exponential growth phase, add 0.25% trypsin digestion solution, digest to detach the adherent cells, and count (1-4) × 10⁻⁶ cells. 5 Cells were prepared as a suspension at a density of 1 cell per mL.

[0141] The cell suspension was seeded into a 12-well plate and incubated in a constant temperature CO2 incubator for 24 hours.

[0142] Except for the control group which was given PBS, each of the other wells was given FFA to induce modeling. Then, different concentrations of the test samples were added to the experimental groups and incubated in a constant temperature CO2 incubator for 48 hours.

[0143] Collect cells, centrifuge to obtain cell pellet, add RIPA lysis buffer, vortex three times (30s / time, 3min interval), and centrifuge at 12000rpm for 10min. Collect the supernatant, perform the ELISA procedure according to the ELISA instructions, and use the BCA protein quantification kit to determine the total protein concentration in the supernatant and calculate the acetyl-CoA carboxylase content.

[0144] 4.6 Experimental Results

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

[0146] The effects 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 acetyl-CoA carboxylase content in the model group was significantly increased. Compared with the model group, the experimental groups all showed a significant decrease in acetyl-CoA carboxylase content, indicating that the peptide disclosed in this paper can inhibit the expression of acetyl-CoA carboxylase, thereby inhibiting lipid synthesis.

[0147] Therefore, it can be seen that the peptide disclosed herein can reduce the content of acetyl-CoA carboxylase, inhibit the synthesis or secretion of skin oil, slow down oil deposition, reduce the occurrence of pimples or acne, and also help the skin recover after pimples or acne occur, thus having the effects of oil control, acne removal, and repair.

[0148] Example 5: Emulsion containing peptide (5)

[0149]

[0150] Preparation method: According to the formula dosage, heat all materials of phase A in the oil phase pot to 75-80℃ and stir until completely dissolved; add all materials of phase B to the emulsification pot, stir and heat to 80-85℃ until completely dissolved; draw 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 phase C and phase D materials and continue stirring; cool to 35-40℃, add phase E materials, and stir for 10-15 minutes to obtain the final product.

[0151] Example 6: Essence containing peptide (6)

[0152]

[0153] Preparation method: According to the formula dosage, add all materials of phase A to the mixing pot, stir and heat to 80-85℃; mix all materials of phase B evenly until there are no powder particles, add to the mixing pot, and continue stirring for 10-15 minutes; start cooling down, cool down to 60-65℃, add materials of phase C; cool down to 35-40℃, add materials of phases D and E, stir for 10-15 minutes, and the product is obtained.

[0154] Example 7 Cream containing peptide (1)

[0155]

[0156] Preparation method: According to the formula dosage, heat the D phase material in a suitable container to 55-60℃ until completely dissolved, and set aside; add the A phase to a stirring pot and heat to 80-85℃; add the B phase materials to the oil phase pot and heat to 75-80℃ until completely dissolved and transparent; add the B phase to the A phase, turn on the vacuum, homogenize for 5 minutes, maintain stirring, and keep warm for 20 minutes; start cooling, cool to 60-65℃, add the C phase and pre-dissolved D phase materials, homogenize for 2 minutes; cool to 35-40℃, add the E phase material, and stir for 10-15 minutes to obtain the final product.

[0157] In this disclosure, relational terms such as "first" and "second" are used merely to distinguish one entity or operation from another, and do not necessarily require or imply any such actual relationship or order between these entities or operations. Furthermore, the terms "comprising," "including," or any other variations thereof are intended to cover non-exclusive inclusion, such that a process, method, article, or terminal device that comprises a list of elements includes not only those elements but also other elements not expressly listed, or elements inherent to such a process, method, article, or terminal device. Without further limitation, an element defined by the phrase "comprising one..." does not exclude the presence of other identical elements in the process, method, article, or terminal device that includes said element.

[0158] While specific embodiments of this disclosure have been described for illustrative purposes, various modifications or alterations can be made by those skilled in the art without departing from the spirit and scope of this disclosure. All such modifications or alterations should fall within the scope of the appended claims.

Claims

1. The peptide represented by formula (I), or a stereoisomer thereof, a mixture thereof, or a salt thereof, R1-Val-Ser-Gly-Leu-Thr-Pro-R2(I) In formula (I), R1 is selected from H or R3-CO-, where R3 is selected from substituted or unsubstituted alkyl or substituted or unsubstituted alkenyl groups; R2 is selected from: -NR4R5 or -OR4, wherein each R4 and R5 is independently selected from: H, substituted or unsubstituted alkyl, substituted or unsubstituted alkenyl.

2. The peptide, or its stereoisomer, or mixture thereof, or salt thereof, of formula (I) according to claim 1, characterized in that, 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. Optionally, 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. Optionally, R1 is selected from H, acetyl, lauroyl, myristoyl, or palmitoyl; R2 is -OH or -NH2.

3. The peptide, or its stereoisomer, or a mixture thereof, or a salt thereof, of formula (I) 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.

4. The peptide, stereoisomer thereof, mixture thereof, or salt thereof of formula (I) according to any one of claims 1-3, characterized in that, The salt comprises a metal salt of the peptide represented by formula (I), wherein the metal comprises: lithium, sodium, potassium, calcium, magnesium, manganese, copper, zinc or aluminum; Alternatively, the salt may comprise a salt formed from a peptide of formula (I) and an organic base, wherein the organic base comprises: ethylamine, diethylamine, ethylenediamine, ethanolamine, diethanolamine, arginine, lysine, histidine, or piperazine; Alternatively, the salt may comprise a salt formed from a peptide of formula (I) and an inorganic or 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, pyric acid, or gluconic acid; and the inorganic acid includes hydrochloric acid, sulfuric acid, boric acid, or carbonic acid.

5. A composition, characterized in that, It includes an effective amount of the peptide of formula (I) as described in any one of claims 1-4, or a stereoisomer thereof, a mixture thereof, or a salt thereof, as well as at least one excipient and optional adjuvant.

6. A delivery system or sustained-release system, characterized in that, Contains an effective amount of the peptide of formula (I) as described in any one of claims 1-4, or a stereoisomer thereof, a mixture thereof, a salt thereof, or the composition of claim 5.

7. The delivery system or sustained-release system according to claim 6, characterized in that, The delivery system or sustained-release system includes: liposomes, oil bodies, alcohol bodies, millimeter capsules, micrometer capsules, nanocapsules, nanostructured lipid carriers, sponges, inclusion complexes, lipid vesicles, micelles, millimeter spheres, micrometer spheres, nanospheres, lipid spheres, micrometer emulsions, nanoemulsions, millimeter particles, micrometer particles, or nanoparticles.

8. A cosmetic product, characterized in that, Contains an effective amount of the peptide of formula (I) as described in any one of claims 1-4, or a stereoisomer thereof, a mixture thereof, or a salt thereof, or the composition of claim 5, or the delivery system or sustained-release system of claim 6 or 7.

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

10. Use of the peptide of formula (I) according to any one of claims 1-4, or a stereoisomer thereof, a mixture thereof, a salt thereof, or a composition according to claim 5, or a delivery system or sustained-release system according to claim 6 or 7, in the preparation of a composition for the care or treatment of skin.

11. Use of the peptide of formula (I) according to any one of claims 1-4, or a stereoisomer thereof, a mixture thereof, a salt thereof, or a composition according to claim 5, or a delivery system or sustained-release system according to claim 6 or 7, in the preparation of a composition for oil control, acne treatment or repair.

12. Use of the peptide of formula (I) according to any one of claims 1-4, or a stereoisomer thereof, a mixture thereof, a salt thereof, or a composition according to claim 5, or a delivery system or sustained-release system according to claim 6 or 7, in the preparation of a composition for inhibiting the synthesis or secretion of sebum in the skin, or in the preparation of a composition for reducing the content of acetyl-CoA carboxylase.

13. Use of the peptide of formula (I) according to any one of claims 1-4, or a stereoisomer thereof, a mixture thereof, a salt thereof, or a composition according to claim 5, or a delivery system or sustained-release system according to claim 6 or 7 in the preparation of cosmetics.

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