Cyclic peptide compound and application thereof

Abstract

The invention relates to a cyclic peptide compound and an application thereof, and provides a cyclic peptide compound which is shown in a formula I, is novel in structure and has the effect equivalent to or even better than that of GHK tripeptide, and an application of the cyclic peptide compound in a dermatological or cosmetic composition for stimulating hair growth or resisting skin aging. And the formula I is * AA1-AA2-GLY-HIS-LYS-AA4-AA3 *, and the formula I is shown in the description.

Description

Cyclic peptide compounds and their applications

[0001] Citation of Related Applications

[0002] This application claims priority to Chinese patent application CN202311247312.8 filed on September 25, 2023 and Chinese patent application CN202411310856.9 filed on September 19, 2024, the contents of which are incorporated by reference into this application in their entirety and for all purposes. Technical Field

[0003] The present invention relates to the field of biomedicine or cosmetic products, and in particular to cyclic peptide compounds and applications thereof. Background Art

[0004] GHK is a tripeptide molecule, also known as tripeptide or tripeptide-1. Its sequence is glycyl-histidyl-lysine, and it's a product of collagen hydrolysis. It's naturally present in human blood, saliva, and urine. GHK is known for its ability to boost the production of type I collagen fragments. Its mechanism of action can be simply understood as when collagen naturally breaks down in the skin, the resulting peptide fragments signal the skin to get to work and produce some beautiful new collagen.

[0005] GHK has a high affinity for divalent copper ions and can spontaneously combine with them to form a complex GHK-Cu, which is also called blue copper peptide because its aqueous solution is blue. It is the main form in which GHK exerts its effects in the body. Its functions mainly include: (1) chemotaxis of repair cells, such as macrophages, mast cells, and capillary cells; (2) anti-inflammatory effects (inhibiting free radicals, thromboxane formation, transforming growth factor β-1, tumor necrosis factor α and protein glycation, while increasing superoxide dismutase, dilating blood vessels, blocking ultraviolet damage to skin keratinocytes and improving the recovery of fibroblasts after X-ray treatment); (3) increasing the protein synthesis of collagen, elastin, hyaluronic acid, metalloproteinases, antiproteinases, vascular endothelial growth factor, fibroblast growth factor 2, nerve growth factor, neutrophil 3 and 4, and erythropoietin; (4) increasing the proliferation of fibroblasts and keratinocytes; nerve growth, angiogenesis, and hair follicle size. Controlled studies on aging skin have shown that GHK-Cu can tighten the skin, improve elasticity and firmness, reduce fine lines, wrinkles, photodamage and hyperpigmentation, and promote hair growth and improve the success rate of hair transplantation.

[0006] Summary of the Invention

[0007] The present invention provides a cyclic peptide compound with a novel structure and efficacy comparable to or even better than that of a GHK tripeptide, and its use in a dermatological or cosmetic composition for stimulating hair growth or anti-aging of the skin.

[0008] To this end, in the first aspect of the present invention, the present invention provides a cyclic peptide compound represented by Formula I or a stereoisomer thereof, *AA1-AA2-GLY-HIS-LYS-AA4-AA3* Formula I

[0009] in:

[0010] AA1 is an amino acid residue;

[0011] AA2 is absent or is an amino acid residue;

[0012] AA3 is an amino acid residue;

[0013] AA4 is absent or is an amino acid residue;

[0014] * indicates that the N-terminus and C-terminus of formula I are connected to form a cyclic peptide;

[0015] The amino acid is a natural amino acid or a non-natural amino acid;

[0016] The amino acids are D-type or L-type.

[0017] It should be noted that the left side of Formula I is the N-terminus, and the right side is the C-terminus.

[0018] In some embodiments, the amino acids are in the L-form.

[0019] In some embodiments, the amino acid is a natural amino acid.

[0020] In some embodiments, the amino acid is a hydrophobic natural amino acid.

[0021] In some embodiments, the amino acid is selected from alanine (ALA; A), glycine (GLY; G), isoleucine (ILE; I), leucine (LEU; L), methionine (MET; M), phenylalanine (PHE; F), proline (PRO; P), tryptophan (TRP; W), and valine (VAL; V).

[0022] In some embodiments, AA1 is selected from the group consisting of a phenylalanine residue, an alanine residue, a methionine residue, a leucine residue, a tryptophan residue, an isoleucine residue, a proline residue, a glycine residue, and a valine residue.

[0023] In some embodiments, AA1 is selected from an alanine residue, a methionine residue, a phenylalanine residue, a leucine residue, a tryptophan residue, an isoleucine residue, a proline residue, a glycine residue.

[0024] In some embodiments, AA1 is selected from a methionine residue, an alanine residue, a phenylalanine residue, a glycine residue, a proline residue.

[0025] In some embodiments, AA2 is an amino acid residue.

[0026] In some embodiments, AA2 is selected from the group consisting of a phenylalanine residue, an alanine residue, a methionine residue, a leucine residue, a tryptophan residue, an isoleucine residue, a proline residue, a glycine residue, and a valine residue.

[0027] In some embodiments, AA2 is selected from a valine residue, a proline residue, a leucine residue, an alanine residue, an isoleucine residue, a methionine residue, a tryptophan residue.

[0028] In some embodiments, AA2 is selected from a proline residue, a leucine residue, a valine residue, an isoleucine residue, a tryptophan residue.

[0029] In some embodiments, AA3 is selected from a phenylalanine residue, an alanine residue, a methionine residue, a leucine residue, a tryptophan residue, an isoleucine residue, a proline residue, a glycine residue, a valine residue.

[0030] In some embodiments, AA3 is selected from a phenylalanine residue, an alanine residue, a methionine residue, a leucine residue, a tryptophan residue, an isoleucine residue, a glycine residue, a valine residue.

[0031] In some embodiments, AA3 is selected from a phenylalanine residue, an alanine residue, a methionine residue, a leucine residue, an isoleucine residue, a valine residue.

[0032] In some embodiments, AA3 is selected from an isoleucine residue, a leucine residue, a phenylalanine residue, a methionine residue, a valine residue.

[0033] In some embodiments, AA4 is absent.

[0034] In some embodiments, the cyclic peptide compound has a structure shown in Formula I-1, *AA3-AA1-AA2-GLY-HIS-LYS* Formula I-1

[0035] in:

[0036] AA1 is selected from the group consisting of a phenylalanine residue, an alanine residue, a methionine residue, a leucine residue, a tryptophan residue, an isoleucine residue, a proline residue, a glycine residue, and a valine residue;

[0037] and / or

[0038] AA2 is selected from the group consisting of a phenylalanine residue, an alanine residue, a methionine residue, a leucine residue, a tryptophan residue, an isoleucine residue, a proline residue, a glycine residue, and a valine residue;

[0039] and / or

[0040] AA3 is selected from the group consisting of a phenylalanine residue, an alanine residue, a methionine residue, a leucine residue, a tryptophan residue, an isoleucine residue, a proline residue, a glycine residue, and a valine residue;

[0041] * indicates that the N-terminus and C-terminus of formula I-1 are connected to form a cyclic peptide;

[0042] The amino acids are D-type or L-type.

[0043] It should be noted that the left side of formula I-1 is the N-terminus, and the right side is the C-terminus.

[0044] In some embodiments, the amino acids are in the L-form.

[0045] In some embodiments, AA1 is selected from the group consisting of a phenylalanine residue, an alanine residue, a methionine residue, a leucine residue, a tryptophan residue, an isoleucine residue, a proline residue, and a glycine residue.

[0046] In some embodiments, AA1 is selected from a methionine residue, an alanine residue, a phenylalanine residue, a glycine residue, a proline residue.

[0047] In some embodiments, AA2 is selected from an alanine residue, a methionine residue, a leucine residue, a tryptophan residue, an isoleucine residue, a proline residue, a valine residue.

[0048] In some embodiments, AA2 is selected from a proline residue, a leucine residue, a valine residue, an isoleucine residue, a tryptophan residue.

[0049] In some embodiments, AA3 is selected from a phenylalanine residue, an alanine residue, a methionine residue, a leucine residue, a tryptophan residue, an isoleucine residue, a glycine residue, a valine residue.

[0050] In some embodiments, AA3 is selected from a phenylalanine residue, an alanine residue, a methionine residue, a leucine residue, an isoleucine residue, a valine residue.

[0051] In some embodiments, AA3 is selected from an isoleucine residue, a leucine residue, a phenylalanine residue, a methionine residue, a valine residue.

[0052] In some embodiments, the structural formula of the cyclic peptide compound is shown in Table A.

[0053] Table A: Cyclic peptide compound structure (all amino acids are L-type)

[0054] In some embodiments, the structural formula of the cyclic peptide compound is shown in Table B.

[0055] Table B: Preferred cyclic peptide compound structural formula (all amino acids are L-type)

[0056] In some embodiments, the structural formula of the cyclic peptide compound is shown in Table C.

[0057] Table C: More preferred cyclic peptide compound structural formula (all amino acids are L-type)

[0058] In some embodiments, the structural formula of the cyclic peptide compound is shown in Table D.

[0059] Table D: The most preferred cyclic peptide compound structure (all amino acids are L-type)

[0060] In the structural formulas of the cyclic peptide compounds shown in Table A, Table B, Table C and Table D, the left amino acid forms a ring with the right amino acid through a peptide bond.

[0061] In the second aspect of the present invention, the present invention provides a pharmaceutical composition comprising the aforementioned cyclic peptide compound or a stereoisomer thereof.

[0062] In some embodiments, the pharmaceutical composition further comprises at least one carrier and / or excipient.

[0063] In the third aspect of the present invention, the present invention provides a cosmetic composition (such as a skin care product, a cosmetic, a hair growth liquid, etc.), which comprises the aforementioned cyclic peptide compound or a stereoisomer thereof.

[0064] In some embodiments, the cosmetic composition further comprises a functional substance.

[0065] In some embodiments, the functional substance is selected from one or more of a skin conditioner, an antioxidant, a surfactant, a moisturizer, a preservative, a chelating agent, and a fragrance.

[0066] In a fourth aspect of the present invention, the present invention provides the use of the aforementioned cyclic peptide compound or its stereoisomer or the aforementioned pharmaceutical composition in the preparation of a drug, wherein the drug is used for one or more selected from the following (1)-(7):

[0067] (1) Delay skin aging;

[0068] (2) Reduce skin damage;

[0069] (3) Accelerate skin wound healing;

[0070] (4) Reduce hyperpigmentation;

[0071] (5) stimulate hair growth;

[0072] (6) Improve the success rate of hair transplantation;

[0073] (7) Prevent hair loss.

[0074] In a fifth aspect of the present invention, the present invention provides the aforementioned cyclic peptide compound or its stereoisomer or the aforementioned pharmaceutical composition, which is used for one or more uses selected from the following (1)-(7):

[0075] (1) Delay skin aging;

[0076] (2) Reduce skin damage;

[0077] (3) Accelerate skin wound healing;

[0078] (4) Reduce hyperpigmentation;

[0079] (5) stimulate hair growth;

[0080] (6) Improve the success rate of hair transplantation;

[0081] (7) Prevent hair loss.

[0082] In a sixth aspect of the present invention, the present invention provides a method selected from one or more of the following (1)-(7), comprising: administering an effective amount of the aforementioned cyclic peptide compound or its stereoisomer or the aforementioned pharmaceutical composition to a subject in need thereof,

[0083] (1) Methods for delaying skin aging;

[0084] (2) Methods to reduce skin damage;

[0085] (3) Methods for accelerating skin wound healing;

[0086] (4) methods to reduce hyperpigmentation;

[0087] (5) Methods of stimulating hair growth;

[0088] (6) Methods to improve the success rate of hair transplantation;

[0089] (7) Methods to prevent hair loss.

[0090] In a seventh aspect of the present invention, the present invention provides the use of the aforementioned cyclic peptide compound or its stereoisomer or the aforementioned cosmetic composition for one or more selected from the following (1)-(17), wherein the use is a non-therapeutic use:

[0091] (1) Anti-aging of the skin;

[0092] (2) Anti-wrinkle and reduce fine lines;

[0093] (3) Improve the mechanical properties of the skin: firmness, skin tone, elasticity and / or flexibility;

[0094] (4) Improve skin density (reconstruction effect);

[0095] (5) Increase skin volume (volumizing effect);

[0096] (6) Counteract the appearance of stretch marks;

[0097] (7) Reduce skin damage;

[0098] (8) Accelerate skin wound healing;

[0099] (9) Anti-spot;

[0100] (10) Improve skin homogeneity and / or radiance;

[0101] (11) Reduce skin pigmentation;

[0102] (12)Moisturizing;

[0103] (13) Stimulate hair growth;

[0104] (14) Improve the success rate of hair transplantation;

[0105] (15) Prevent hair loss;

[0106] (16) Local beauty treatment of eyelashes;

[0107] (17) Increase hair width, density and / or length.

[0108] In an eighth aspect of the present invention, the present invention provides a method selected from one or more of the following (1)-(17), comprising: administering an effective amount of the aforementioned cyclic peptide compound or its stereoisomer or the aforementioned cosmetic composition to a subject in need thereof,

[0109] (1) Methods for skin anti-aging;

[0110] (2) Methods for anti-wrinkle and fine line reduction;

[0111] (3) methods to improve the mechanical properties of the skin: firmness, skin tone, elasticity, and / or flexibility;

[0112] (4) methods to increase skin density (reconstruction effect);

[0113] (5) Methods for increasing skin volume (volumizing effect);

[0114] (6) Methods to combat the appearance of stretch marks;

[0115] (7) Methods to reduce skin damage;

[0116] (8) Methods for accelerating skin wound healing;

[0117] (9) Anti-spot methods;

[0118] (10) Methods for improving skin homogeneity and / or radiance;

[0119] (11) Methods for reducing skin pigmentation;

[0120] (12) Moisturizing methods;

[0121] (13) Methods of stimulating hair growth;

[0122] (14) Methods to improve the success rate of hair transplantation;

[0123] (15) Methods for preventing hair loss;

[0124] (16) Local cosmetic methods for eyelashes;

[0125] (17) Methods for increasing hair width, density and / or length.

[0126] In the ninth aspect of the present invention, the present invention provides the use of the aforementioned cyclic peptide compound or its stereoisomer or the aforementioned cosmetic composition for one or more selected from the following (1)-(17):

[0127] (1) Anti-aging of the skin;

[0128] (2) Anti-wrinkle and reduce fine lines;

[0129] (3) Improve the mechanical properties of the skin: firmness, skin tone, elasticity and / or flexibility;

[0130] (4) Improve skin density (reconstruction effect);

[0131] (5) Increase skin volume (volumizing effect);

[0132] (6) Counteract the appearance of stretch marks;

[0133] (7) Reduce skin damage;

[0134] (8) Accelerate skin wound healing;

[0135] (9) Anti-spot;

[0136] (10) Improve skin homogeneity and / or radiance;

[0137] (11) Reduce skin pigmentation;

[0138] (12)Moisturizing;

[0139] (13) Stimulate hair growth;

[0140] (14) Improve the success rate of hair transplantation;

[0141] (15) Prevent hair loss;

[0142] (16) Local beauty treatment of eyelashes;

[0143] (17) Increase hair width, density and / or length. Beneficial effects

[0144] 1. The cyclic peptide compounds of the present invention (e.g., the cyclic peptide compounds in Table A, Table B, Table C, or Table D) or their stereoisomers can achieve anti-wrinkle effects by significantly increasing the content of Collagen I.

[0145] 2. The cyclic peptide compounds of the present invention (e.g., the cyclic peptide compounds in Table A, Table B, Table C, or Table D) or their stereoisomers can significantly increase the hyaluronic acid (HA) content, thereby achieving a firming effect.

[0146] 3. The cyclic peptide compounds of the present invention (such as the cyclic peptide compounds in Table A, Table B, Table C or Table D) or their stereoisomers can significantly increase the content of elastin, thereby achieving a firming effect.

[0147] 4. The cyclic peptide compounds of the present invention (e.g., the cyclic peptide compounds in Table A, Table B, Table C, or Table D) or their stereoisomers can significantly increase the content of Collagen I, hyaluronic acid (HA), and elastin, thereby achieving anti-wrinkle and firming effects, and having excellent medical and cosmetic effects.

[0148] 5. The cyclic peptide compounds of the present invention (e.g., the cyclic peptide compounds in Table A, Table B, Table C, or Table D) do not show significant cytotoxicity (e.g., within a concentration range of 5 μM) and have good application prospects in medicine and cosmetics.

[0149] 6. The cyclic peptide compounds of the present invention (e.g., the cyclic peptide compounds in Table A, Table B, Table C, or Table D) or their stereoisomers, when used as medicine, have one or more beneficial effects selected from the following:

[0150] (1) Delay skin aging;

[0151] (2) Reduce skin damage;

[0152] (3) Accelerate skin wound healing;

[0153] (4) Reduce hyperpigmentation;

[0154] (5) stimulate hair growth;

[0155] (6) Improve the success rate of hair transplantation;

[0156] (7) Prevent hair loss.

[0157] 7. The cyclic peptide compounds of the present invention (e.g., the cyclic peptide compounds in Table A, Table B, Table C, or Table D) or their stereoisomers, when used for non-therapeutic cosmetic purposes, have one or more beneficial effects selected from the following:

[0158] (1) Anti-aging of the skin;

[0159] (2) Anti-wrinkle and reduce fine lines;

[0160] (3) Improve the mechanical properties of the skin: firmness, skin tone, elasticity and / or flexibility;

[0161] (4) Improve skin density (reconstruction effect);

[0162] (5) Increase skin volume (volumizing effect);

[0163] (6) Counteract the appearance of stretch marks;

[0164] (7) Reduce skin damage;

[0165] (8) Accelerate skin wound healing;

[0166] (9) Anti-spot;

[0167] (10) Improve skin homogeneity and / or radiance;

[0168] (11) Reduce skin pigmentation;

[0169] (12)Moisturizing;

[0170] (13) Stimulate hair growth;

[0171] (14) Improve the success rate of hair transplantation;

[0172] (15) Prevent hair loss;

[0173] (16) Local beauty treatment of eyelashes;

[0174] (17) Increase hair width, density and / or length.

[0175] 8. The cyclic peptide compounds of the present invention (e.g., the cyclic peptide compounds in Table A, Table B, Table C, or Table D) or their stereoisomers, when used as cosmetics, have one or more of the following beneficial effects:

[0176] (1) Anti-aging of the skin;

[0177] (2) Anti-wrinkle and reduce fine lines;

[0178] (3) Improve the mechanical properties of the skin: firmness, skin tone, elasticity and / or flexibility;

[0179] (4) Improve skin density (reconstruction effect);

[0180] (5) Increase skin volume (volumizing effect);

[0181] (6) Counteract the appearance of stretch marks;

[0182] (7) Reduce skin damage;

[0183] (8) Accelerate skin wound healing;

[0184] (9) Anti-spot;

[0185] (10) Improve skin homogeneity and / or radiance;

[0186] (11) Reduce skin pigmentation;

[0187] (12)Moisturizing;

[0188] (13) Stimulate hair growth;

[0189] (14) Improve the success rate of hair transplantation;

[0190] (15) Prevent hair loss;

[0191] (16) Local beauty treatment of eyelashes;

[0192] (17) Increase hair width, density and / or length.

[0193] 9. The efficacy of the cyclic peptide compounds of the present invention (such as the cyclic peptide compounds in Table A, Table B, Table C or Table D) or their stereoisomers in the above eight aspects is comparable to or even better than that of the GHK tripeptide. DETAILED DESCRIPTION

[0194] The embodiments of the present invention will be described in detail below with reference to the examples, but it will be understood by those skilled in the art that the following examples are merely illustrative of the present invention and should not be construed as limiting the scope of the invention. Where specific conditions are not specified in the examples, the methods were performed according to conventional conditions or the conditions recommended by the manufacturer. Where the manufacturers of the reagents or instruments are not specified, they are all conventional products that can be obtained commercially.

[0195] Unless otherwise indicated, scientific and technical terms used herein have the meanings commonly understood by those skilled in the art. Furthermore, terms and laboratory procedures related to protein and nucleic acid chemistry, molecular biology, cell and tissue culture, microbiology, and immunology used herein are those widely used in the respective fields and are standard procedures. To facilitate a better understanding of the present invention, definitions and explanations of relevant terms are provided below.

[0196] The term "stereoisomer" refers to a compound with the same molecular formula in which the atoms or substituents are connected in the same order but arranged in space in different ways, which is a phenomenon of isomerism in the field of organic chemistry.

[0197] The term "absent" indicates that the groups on either side are directly connected. For example, in the cyclic peptide compound of Formula I, if AA4 is absent, the structural formula of the cyclic peptide compound of Formula I becomes *AA1-AA2-GLY-HIS-LYS-AA3*.

[0198] The term "carrier" refers to a system that can change the way a drug enters the human body and its distribution within the body, control the rate of drug release, and deliver the drug to a targeted organ. Drug carrier release and targeting systems can reduce drug degradation and loss, reduce side effects, and improve bioavailability. For example, polymer surfactants that can be used as carriers can self-assemble to form various forms of aggregates due to their unique amphiphilic structure. Preferred examples include micelles, microemulsions, gels, liquid crystals, and vesicles. These aggregates have the ability to encapsulate drug molecules while also having good membrane permeability, making them excellent drug carriers.

[0199] The term "excipient" refers to an additive other than the main drug in a pharmaceutical preparation, which may also be called an excipient. Excipients include, but are not limited to: ion exchangers, alumina, aluminum stearate, lecithin, serum proteins such as human albumin, buffer substances such as phosphates, glycerol, sorbic acid, potassium sorbate, partial glyceride mixtures of saturated vegetable fatty acids, water, salts or electrolytes, such as protamine sulfate, disodium hydrogen phosphate, potassium hydrogen phosphate, sodium chloride, zinc salts, colloidal silica, magnesium trisilicate, polyvinyl pyrrolidone, cellulosic substances, polyethylene glycol, sodium carboxymethyl cellulose, polyacrylates, beeswax, lanolin, etc.

[0200] The term "skin conditioning agent" includes, but is not limited to, hydroquinone, kojic acid, arbutin, ellagic acid, azelaic acid, azeloyl diglycine, L-ascorbic acid and its derivatives, licorice extract, magnolia officinalis fruit extract, hydrangea extract, alkoxysalicylic acid, knotweed extract, sophora japonica flower extract, turmeric extract, mulberry bark extract, emblica fruit extract, pea extract, aloin, and the like.

[0201] The term "antioxidant" includes, but is not limited to, butylated hydroxytoluene, butylated hydroxyanisole, propyl gallate, octyl gallate, lauryl gallate, t-butylhydroquinone, tocopherol, ascorbyl palmitate, isopropyl citrate mixture, monoglyceride citrate, anoxomer, sodium sulfite, sodium bisulfite, sodium metabisulfite, cysteine, isoascorbic acid, thiourea, dilauryl thiomalonate, nordihydroguaiaretic acid, lipoic acid, alpha-tocopheryl acetate, carotenoids, emblica tannins, ascorbic acid, glutathione, coenzyme Q10, and the like.

[0202] The term "surfactant" may include anionic surfactants, cationic surfactants, nonionic surfactants, amphoteric surfactants, natural surfactants, fluorine-containing surfactants, and the like.

[0203] The term "humectant" includes, but is not limited to, glycerin, propylene glycol, butylene glycol, sorbitol, sorbitol polyether-20, amide moisturizers, glyceryl polyether moisturizers, lactic acid and sodium lactate, sodium pyrrolidone carboxylate, hydroxyethyl urea, erythritol, glyceryl polyether-5 lactate, sodium hyaluronate, saccharide isomers, D-panthenol, polyethylene glycol, diglycerin, methacryloyloxyethyl phosphorylcholine, etc.

[0204] The term "preservative" includes but is not limited to hydroxyphenyl esters and their salts, formaldehyde and formaldehyde donors, isothiazolinones, phenolic preservatives, acid preservatives, halide preservatives, quaternary ammonium compounds, alcohols, commonly used compound preservatives and natural preservatives, etc.

[0205] The term "chelating agent" may be selected from ethylenediaminetetraacetic acid and its sodium salt, glycine, citric acid, succinic acid and the like.

[0206] The term "essence" can specifically be sandalwood, jasmine, rose, sweet orange, vanilla essence, etc. Essence can also include essential oils, specifically lime peel oil, petitgrain leaf / twig oil, geranium oil, Damascus rose flower oil, etc.

[0207] The term "treat" generally refers to obtaining a desired pharmacological and / or physiological effect. The effect can be prophylactic, in terms of completely or partially preventing a disease or its symptoms; and / or therapeutic, in terms of partially or completely stabilizing or curing a disease and / or causing side effects due to the disease. As used herein, "treat" encompasses any treatment of a disease in a patient, including: (a) preventing the onset of a disease or symptom in a patient who is susceptible to the disease or symptom but has not yet been diagnosed with the disease; (b) suppressing the symptoms of a disease, i.e., arresting its development; or (c) relieving the symptoms of a disease, i.e., causing the disease or symptom to regress.

[0208] The term "subject" includes humans and non-human animals. In the present invention, the term "non-human animal" includes all vertebrates, such as non-mammals (e.g., birds, amphibians, reptiles) and mammals, such as non-human primates, livestock and / or domesticated animals (e.g., sheep, dogs, cats, cows, pigs, etc.).

[0209] The term "effective amount" refers to an amount effective to achieve the desired effect at the necessary dosage and time. The "effective amount" of the present invention may vary depending on factors such as individual skin / hair condition, age, sex and weight.

[0210] In the present invention, the 20 conventional amino acids and their abbreviations follow conventional usage. See Immunology—A Synthesis (2nd edition, ES Golub and DR Gren, Eds., Sinauer Associates, Sunderland, Mass. (1991)), which is incorporated herein by reference.

[0211] It is well known to those skilled in the art that the term "amino acid residue" refers to the incomplete amino acid structure remaining after the amino group of an amino acid loses a hydrogen and the carboxyl group loses a hydroxyl, and has an amino terminus (N-terminus) and a carboxyl terminus (C-terminus).

[0212] In the cyclic peptide compounds of the present invention (including but not limited to the cyclic peptide compounds in Table A, Table B, Table C, and Table D), unless otherwise specified, the default is from left to right, N-terminus to C-terminus, and the left N-terminus is connected to the right C-terminus to form an NC cyclic peptide.

[0213] In addition, the abbreviations in the examples have conventional meanings known in the art and are listed as follows:

[0214] AcOH, Acetic Acid, Acetic Acid

[0215] ACN, Acetonitrile, acetonitrile

[0216] DCM, Dichloromethane (Methylene chloride)

[0217] DMF, N,N-Dimethylformamide, N,N-dimethylformamide

[0218] DIEA, N,N-Diisopropylethylamine, N,N-Diisopropylethylamine

[0219] Fmoc, 9-fluorenylmethyloxycarbonyl, 9-fluorenylmethyloxycarbonyl

[0220] HOBT, 1-Hydroxybenzotriazole

[0221] Piperidine

[0222] PyBOP, Benzotriazol-1-yl-oxytripyrrolidinophosphonium hexafluorophosphate

[0223] Resin

[0224] TBTU, O-(Benzotriazol-1-yl)-N,N,N',N'-tetramethyluronium tetrafluoroborate

[0225] TFA, Trifluoroacetic acid

[0226] TFE, 2,2,2-Trifluoroethanol

[0227] TIS, Triisopropylsilane, triisopropylsilane

[0228] The present invention will be further explained below in conjunction with specific examples, but they should not be construed as limiting the present invention in any way. In addition, unless otherwise specified, all raw materials or reagents can be purchased commercially.

[0229] Example 1: Synthesis and identification of cyclic peptide compounds

[0230] 1. Synthesis of cyclic peptide compounds

[0231] Taking the cyclic peptide compound shown in SEQ ID NO: 52 as an example, starting with lysine, amino acids are condensed sequentially from the C-terminus to the N-terminus (from right to left).

[0232] Step 1: Solid-phase synthesis

[0233] Weigh Fmoc-Lys(Boc)-2-Cl Resin (S = 0.32 mmol / g) and place it in a glass reaction column. Swell it with DCM for 30 minutes, then remove the DCM under reduced pressure. Add a 20% piperidine / DMF solution and react for 20 minutes to remove the Fmoc protecting group. Remove the solution under reduced pressure and wash the resin five times with DMF. Samples were collected for color development and detection to obtain a solid-phase resin with Lys attached (linked via the C-terminus of the Lys).

[0234] Weigh Fmoc-His(Trt)-OH and add it to the resin. Dissolve it in DMF and add DIEA and TBTU. Stir for 60 minutes, remove the solution under reduced pressure, and wash five times with DMF. Sample and colorimetric analysis will yield a solid-phase resin with Lys-His attached (from C-terminus to N-terminus).

[0235] Repeat the above similar process to couple Gly, Pro, Phe, and Ala in sequence, and finally obtain a solid phase resin (from C-terminus to N-terminus) connected to the polypeptide Lys-His-Gly-Pro-Phe-Ala, i.e., a peptide resin.

[0236] Then, the resin was washed three times with DMF, DCM, and methanol in sequence and dried to constant weight.

[0237] Step 2: Peptide resin cleavage

[0238] Preparation of cleavage reagent: Calculate the amount of cleavage reagent based on a volume of 10 mL ± 2 mL of cleavage reagent (AcOH: TFE: DCM = 1:2:7, volume ratio) per 1 g of peptide resin. Specifically, add the required cleavage reagents, DCM, TFE, and AcOH, to the cleavage reaction flask in sequence, maintaining the cleavage reagent temperature at 0-10°C. Stir the cleavage reagent to the peptide resin, allowing the system temperature to stabilize. After stirring and reacting for 2 hours at 25-30°C, filter the cleavage solution.

[0239] Step 3: Dealing with the reaction

[0240] The lysate from step 2 was added to a TBTU / DIEA solution with stirring. After the reaction, the solvent was removed using a rotary evaporator. The amount of the reagent mixture was calculated based on a volume of 10 ml ± 2 ml of a mixture of TFA:H₂O:TIS = 95:2.5:2.5, by volume, per 1 g of peptide resin. The required reagents, H₂O, TFA, and TIS, were then added to the reaction flask in that order. After stirring for 2 hours, the solution was precipitated with 5 times the volume of the concentrate using glacial ether. The precipitate was filtered and dried under reduced pressure at room temperature to obtain the crude product.

[0241] Step 4: Freeze-drying and purification

[0242] The crude product was ground finely, purified water was prepared, and the ground crude product was slowly added under stirring. At the same time, an acetonitrile aqueous solution was added dropwise. After the crude product was added and completely dissolved, it was filtered through a 0.45 μm microporous filter membrane. The crude product was purified using a C-18 column packing preparative column, mobile phase A: 0.1% TFA aqueous solution, mobile phase B: 0.1% TFA acetonitrile solution. Separation and purification were performed at room temperature using an appropriate gradient. The target product was collected, analyzed, tested, and classified. The purity requirement was ≥95%. Unqualified target products were collected and separated and purified again using an appropriate gradient. The qualified main peak was freeze-dried under reduced pressure to obtain a powdered purified polypeptide (SEQ ID NO: 52).

[0243] Those skilled in the art can refer to the above synthesis steps to prepare other cyclic peptide compounds of the present application.

[0244] 2. Identification of cyclic peptide compounds

[0245] Weigh the purified peptide prepared above and dissolve it in a mixture of water and acetonitrile until the solution is clear. Filter and load the sample. HPLC conditions: C18 column, 4.6 μm × 250 mm; mobile phase A: 0.1% TFA in acetonitrile, mobile phase B: 0.1% TFA in water; gradient: 0.01 min 25% A, 75% B → 25 min 50% A, 50% B → 25.01 min 100% A → 30 min stop; detection wavelength: 220 nm; flow rate: 1.0 ml / min; column temperature: 25°C; injection volume: 10 μL.

[0246] The mass spectrometer detector was Agilent-6125B, using an electrospray ionization source (ESI) and positive ion scanning; the mass spectrometer voltage was +4.5 kV; the detector was 1.5 kV; the nebulizer flow rate was 1.5 L / min; the CDL was -20.0 v; the CDL temperature was 250 ° C; the heating module temperature was 200 ° C; the mobile phase was 50% water / 50% acetonitrile; and the flow rate was 0.2 mL / min.

[0247] The mass spectrometry data of some exemplary cyclic peptide compounds are shown in Table 1 below.

[0248] Table 1: Mass spectrometry data of exemplary cyclic peptide compounds

[0249] As shown in Table 1, the preparation method of the present invention can successfully obtain the expected cyclic peptide compound.

[0250] Example 2: Biological activity test of cyclic peptide compounds

[0251] 1. Test method

[0252] A. Fibroblast cytotoxicity test

[0253] Test materials: human fibroblasts (Guangdong Boxi Biotechnology Co., Ltd., batch number Fb20081902), DMEM culture medium (Gibco, 23042301, expiration date 230522), PBS (Solarbio, P1010), MTT (Sigma, M5655), DMSO (Sigma, D4540-1L), CO2 incubator (Thermo, 150I), clean bench (Sujing Antai, SW-CJ-1F), inverted microscope (Olympus, CKX53).

[0254] Test method:

[0255] 1) Cell seeding: After thawing the cells, when the plating rate reaches about 60%, seed the cells into a 96-well plate and incubate in an incubator (37° C., 5% CO 2 ) overnight.

[0256] 2) Experimental Grouping: The experiment was divided into a zero adjustment group, a solvent control group, a positive control group, and a sample group. In the sample group, each sample was set up with 7 concentration gradients, and 3 replicate wells were set up under each concentration gradient.

[0257] 3) Solution preparation: Prepare sample working solutions of different concentrations according to the test concentrations of 0.0781, 0.1563, 0.3125, 0.625, 1.25, 2.5, and 5 μM.

[0258] 4) Dosing: Dosing was performed when the cell plating rate in the 96-well plate reached 50% to 60%. For the solvent control group, 200 μL of culture medium was added to each well; for the positive control group, 200 μL of culture medium containing 10% DMSO was added to each well; for the sample group, 200 μL of culture medium containing the corresponding sample concentration was added to each well; for the zero-adjustment group, no cells were seeded, and only 200 μL of cell culture medium was added. After dosing, the 96-well plate was placed in an incubator (37°C, 5% CO2) and incubated for 24 hours.

[0259] 5) Detection: After incubating the cells for 24 h, discard the supernatant and add MTT working solution (0.5 mg / mL). Incubate at 37°C in the dark for 4 h. After incubation, discard the supernatant and add 150 μL DMSO to each well. Read the OD value at 490 nm.

[0260] 6) Calculation of relative cell viability: Calculate according to the following formula

[0261] The test results of some exemplary cyclic peptide compounds are shown in Table 2.

[0262] Table 2: Test results of exemplary cyclic peptide compounds

[0263] As shown in Table 2, in the fibroblast toxicity test, all cyclic peptide compounds showed no obvious cytotoxicity within the concentration range of 5 μM, and have good application prospects in medicine and cosmetics.

[0264] B. Fibroblast Collagen I (Type I collagen) lifting effect, fibroblast HA (hyaluronic acid) lifting effect, fibroblast elastin (ELN) lifting effect

[0265] Test materials: human fibroblasts (Guangdong Boxi Biotechnology Co., Ltd., batch number Fb20081902), DMEM medium (Gibco, 23042301, expiration date 230522), PBS (Solarbio, P1010), TGF-β1 (Peprotech, 230504), Collagen I ELISA kit (Cusabio, B21025824), Elastin ELISA kit (Abcam, GR3446070-2), HA ELISA kit (RD, P325066). CO2 incubator (Thermo, 150I), clean bench (Sujing Antai, SW-CJ-1F), and microplate reader (BioTek, Epoch).

[0266] Test method:

[0267] 1) Inoculation: After thawing cells, when the plating rate reaches about 60%, inoculate the cells into 6-well plates and incubate in an incubator (37° C., 5% CO 2 ) overnight.

[0268] 2) Solution preparation: Cyclic peptide compound in the sample group, PC2 (Biotinoyl-GHK, ) and PC3 The concentration of each test substance was 1 μM, and the working solution of the test substance was prepared. The blank control group (BC) was only added with cell culture medium.

[0269] 3) Drug administration: When the cell plating rate in the 6-well plate reaches 40% to 60%, drug administration is performed, with a dosage of 2 mL per well. Each group has 3 replicate wells and incubated in an incubator (37° C., 5% CO 2 ) for 24 h.

[0270] 4) ELISA test: ELISA test was performed according to the instructions of the ELISA kit. The test indicators were the fibroblast Collagen I enhancement effect, the fibroblast HA enhancement effect, and the fibroblast elastin enhancement effect.

[0271] The activity test data of some exemplary cyclic peptide compounds are shown in Tables 3A, 3B, and 3C below.

[0272] Table 3A: Data on the effect of some exemplary cyclic peptide compounds on promoting the growth of fibroblasts Collagen I

[0273] As shown in Table 3A, compared with BC, the exemplary cyclic peptide compounds of the present invention, such as the cyclic peptide compounds represented by SEQ ID NOs: 214, 414, 250, 297, 205, 289, 232, 268, 707, 340, and 710, significantly increased the type I collagen (Collagen I) content, indicating that the cyclic peptide compounds of the present invention can achieve anti-wrinkle effects by increasing the content of Collagen I.

[0274] Table 3B: Data on the fibroblast HA enhancement effect of some exemplary cyclic peptide compounds

[0275] As shown in Table 3B, compared with BC, the hyaluronic acid (HA) content of the exemplary cyclic peptide compounds of the present invention, such as the cyclic peptide compounds represented by SEQ ID NOs: 52, 250, 297, 205, 291, 196, 292, 414, 214, and 289, is significantly increased, indicating that the cyclic peptide compounds of the present invention can achieve a firming effect by increasing the HA content.

[0276] Table 3C: Data on the fibroblast elastin-enhancing effects of some exemplary cyclic peptide compounds

[0277] As shown in Table 3C, compared with BC, the cyclic peptide compounds of the present invention, such as the cyclic peptide compounds represented by SEQ ID NOs: 52, 250, 297, 205, 291, 196, 292, 414, 214, 289, 232, 340, and 710, have significantly increased elastin content, indicating that the cyclic peptide compounds of the present invention can achieve a firming effect by increasing elastin content.

[0278] In addition, as shown in Table 3A, Table 3B, and Table 3C, compared with BC, the exemplary cyclic peptide compounds of the present invention, such as the cyclic peptide compounds shown in SEQ ID NO: 414, 250, 205, 292, etc., have significantly increased type I collagen (Collagen I) content, hyaluronic acid (HA) content, and elastin content, indicating that the cyclic peptide compounds of the present invention can achieve firming and anti-wrinkle effects by increasing the Collagen I content, HA content, and elastin content, and have excellent medical and cosmetic effects.

Claims

1. The cyclic peptide compound represented by formula I or its stereoisomer, *AA1-AA2-GLY-HIS-LYS-AA4-AA3* Formula I in: AA1 is an amino acid residue; AA2 is absent or is an amino acid residue; AA3 is an amino acid residue; AA4 is absent or is an amino acid residue; * indicates that the N-terminus and the C-terminus of formula I are connected to form a cyclic peptide; The amino acid is a natural amino acid or a non-natural amino acid; The amino acid is in D-type or L-type, preferably in L-type.

2. The cyclic peptide compound or its stereoisomer according to claim 1, wherein The amino acids are natural amino acids; Preferably, the amino acid is a hydrophobic natural amino acid; More preferably, the amino acid is selected from alanine, glycine, isoleucine, leucine, methionine, phenylalanine, proline, tryptophan, and valine.

3. The cyclic peptide compound or its stereoisomer according to any one of claims 1 to 2, wherein: AA1 is selected from the group consisting of a phenylalanine residue, an alanine residue, a methionine residue, a leucine residue, a tryptophan residue, an isoleucine residue, a proline residue, a glycine residue, and a valine residue; Preferably, AA1 is selected from an alanine residue, a methionine residue, a phenylalanine residue, a leucine residue, a tryptophan residue, an isoleucine residue, a proline residue, a glycine residue; More preferably, AA1 is selected from a methionine residue, an alanine residue, a phenylalanine residue, a glycine residue, a proline residue.

4. The cyclic peptide compound or stereoisomer thereof according to any one of claims 1 to 3, wherein: AA2 is an amino acid residue; Preferably, AA2 is selected from the group consisting of a phenylalanine residue, an alanine residue, a methionine residue, a leucine residue, a tryptophan residue, an isoleucine residue, a proline residue, a glycine residue, and a valine residue; Preferably, AA2 is selected from a valine residue, a proline residue, a leucine residue, an alanine residue, an isoleucine residue, a methionine residue, a tryptophan residue; More preferably, AA2 is selected from a proline residue, a leucine residue, a valine residue, an isoleucine residue, a tryptophan residue.

5. The cyclic peptide compound or stereoisomer thereof according to any one of claims 1 to 4, wherein: AA3 is selected from the group consisting of a phenylalanine residue, an alanine residue, a methionine residue, a leucine residue, a tryptophan residue, an isoleucine residue, a proline residue, a glycine residue, and a valine residue; Preferably, AA3 is selected from a phenylalanine residue, an alanine residue, a methionine residue, a leucine residue, a tryptophan residue, an isoleucine residue, a glycine residue, a valine residue; Preferably, AA3 is selected from a phenylalanine residue, an alanine residue, a methionine residue, a leucine residue, an isoleucine residue, a valine residue; More preferably, AA3 is selected from the group consisting of isoleucine residues, leucine residues, phenylalanine residues, methionine residues, Acid residues, valine residues.

6. The cyclic peptide compound or stereoisomer thereof according to any one of claims 1 to 5, wherein: AA4 does not exist.

7. The cyclic peptide compound or stereoisomer thereof according to any one of claims 1 to 6, wherein: The cyclic peptide compound has a structure shown in Formula I-1, *AA3-AA1-AA2-GLY-HIS-LYS* Formula I-1 in: AA1 is selected from the group consisting of a phenylalanine residue, an alanine residue, a methionine residue, a leucine residue, a tryptophan residue, an isoleucine residue, a proline residue, a glycine residue, and a valine residue; Preferably, AA1 is selected from a phenylalanine residue, an alanine residue, a methionine residue, a leucine residue, a tryptophan residue, an isoleucine residue, a proline residue, a glycine residue; and / or AA2 is selected from the group consisting of a phenylalanine residue, an alanine residue, a methionine residue, a leucine residue, a tryptophan residue, an isoleucine residue, a proline residue, a glycine residue, and a valine residue; Preferably, AA2 is selected from an alanine residue, a methionine residue, a leucine residue, a tryptophan residue, an isoleucine residue, a proline residue, a valine residue; and / or AA3 is selected from the group consisting of a phenylalanine residue, an alanine residue, a methionine residue, a leucine residue, a tryptophan residue, an isoleucine residue, a proline residue, a glycine residue, and a valine residue; Preferably, AA3 is selected from a phenylalanine residue, an alanine residue, a methionine residue, a leucine residue, a tryptophan residue, an isoleucine residue, a glycine residue, a valine residue; More preferably, AA3 is selected from a phenylalanine residue, an alanine residue, a methionine residue, a leucine residue, an isoleucine residue, a valine residue; * indicates that the N-terminus and the C-terminus of formula I-1 are connected to form a cyclic peptide; The amino acid is in D-type or L-type, preferably in L-type.

8. The cyclic peptide compound or its stereoisomer according to claim 7, wherein Preferably, AA1 is selected from a methionine residue, an alanine residue, a phenylalanine residue, a glycine residue, a proline residue; and / or Preferably AA2 is selected from a proline residue, a leucine residue, a valine residue, an isoleucine residue, a tryptophan residue; and / or Preferably, AA3 is selected from an isoleucine residue, a leucine residue, a phenylalanine residue, a methionine residue, a valine residue.

9. The cyclic peptide compound or stereoisomer thereof according to any one of claims 1 to 8, wherein: The structural formula of the cyclic peptide compound is shown in Table A; Preferably, the structural formula of the cyclic peptide compound is as shown in Table B; More preferably, the structural formula of the cyclic peptide compound is as shown in Table C; Most preferably, the structural formula of the cyclic peptide compound is as shown in Table D.

10. A pharmaceutical composition comprising the cyclic peptide compound or a stereoisomer thereof according to any one of claims 1 to 9; Optionally, at least one carrier and / or excipient is also included.

11. A cosmetic composition comprising the cyclic peptide compound or a stereoisomer thereof according to any one of claims 1 to 9; Optionally, further comprising a functional substance; Preferably, the functional substance is selected from one or more of skin conditioners, antioxidants, surfactants, moisturizers, preservatives, chelating agents, and fragrances.

12. Use of the cyclic peptide compound or its stereoisomer according to any one of claims 1 to 9 or the pharmaceutical composition according to claim 10 in the preparation of a drug, wherein the drug is used for one or more selected from the following (1) to (7): (1) Delay skin aging; (2) Reduce skin damage; (3) Accelerate skin wound healing; (4) Reduce hyperpigmentation; (5) Stimulate hair growth; (6) Improve the success rate of hair transplantation; (7) Prevent hair loss.

13. Use of the cyclic peptide compound or its stereoisomer according to any one of claims 1 to 9 or the cosmetic composition according to claim 11 for one or more selected from the following (1) to (17): (1) Anti-aging of the skin; (2) Anti-wrinkle and reduce fine lines; (3) Improve the mechanical properties of the skin: firmness, skin tone, elasticity and / or flexibility; (4) Improve skin density; (5) Increase skin volume; (6) Counteract the appearance of traction marks; (7) Reduce skin damage; (8) Accelerate skin wound healing; (9) Anti-plaque; (10) Improve skin homogeneity and / or radiance; (11) Reduce skin pigmentation; (12) Moisturizing; (13) Stimulate hair growth; (14) Improve the success rate of hair transplantation; (15) Prevent hair loss; (16) Local beauty treatment of eyelashes; (17) Increase hair width, density and / or length.