WO2023184114A1 - 一种多肽及其美容组合物或药用组合物和用途 - Google Patents

一种多肽及其美容组合物或药用组合物和用途 Download PDF

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WO2023184114A1
WO2023184114A1 PCT/CN2022/083491 CN2022083491W WO2023184114A1 WO 2023184114 A1 WO2023184114 A1 WO 2023184114A1 CN 2022083491 W CN2022083491 W CN 2022083491W WO 2023184114 A1 WO2023184114 A1 WO 2023184114A1
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ala
agents
leu
trp
val
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PCT/CN2022/083491
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English (en)
French (fr)
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丁文锋
肖玉
赵文豪
孙新林
陈雪
观富宜
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深圳市维琪科技股份有限公司
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Priority to PCT/CN2022/083491 priority Critical patent/WO2023184114A1/zh
Priority to CN202280029549.6A priority patent/CN117203217A/zh
Publication of WO2023184114A1 publication Critical patent/WO2023184114A1/zh

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    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K38/00—Medicinal preparations containing peptides
    • A61K38/04—Peptides having up to 20 amino acids in a fully defined sequence; Derivatives thereof
    • A61K38/08—Peptides having 5 to 11 amino acids
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P17/00—Drugs for dermatological disorders
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61Q—SPECIFIC USE OF COSMETICS OR SIMILAR TOILETRY PREPARATIONS
    • A61Q19/00—Preparations for care of the skin
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K7/00—Peptides having 5 to 20 amino acids in a fully defined sequence; Derivatives thereof
    • C07K7/04—Linear peptides containing only normal peptide links
    • C07K7/06—Linear peptides containing only normal peptide links having 5 to 11 amino acids

Definitions

  • the present application relates to the technical fields of medicine and cosmetics, in particular to a polypeptide and its cosmetic compositions or pharmaceutical compositions and uses.
  • the most obvious sign of human skin aging is the production of facial wrinkles.
  • the body's nerve activity becomes excessive, and the neurotransmitters that stimulate muscle fibers are uncontrolled and excessively released, thereby dragging the muscle tension of the skin inward. , deepening the primary line on the skin surface and forming wrinkles.
  • fibroblasts that synthesize extracellular matrix and collagen located on tension lines exert specific contractile properties associated with striated muscle in response to muscle contraction.
  • the junction between the nerve and striated muscle constitutes the neuromuscular plate, upstream of which is the motor neuron afferent pathway.
  • the vesicles must first fuse with the presynaptic membrane to complete the exocytosis process and release acetylcholine.
  • Membrane fusion is mediated by the N-ethylmaleimide-sensitive factor attachment protein receptor (SNARE) complex.
  • the SNARE complex is a four-helix bundle structure, consisting of synaptic vesicle-associated membrane protein (VAMP2) located on the vesicle membrane, synapse-associated protein (SNAP25) located on the presynaptic membrane, and syntaxin. constitute.
  • Synaptophysin is a synaptic vesicle transmembrane glycoprotein that exists in presynaptic vesicles of neurons and neurosecretory granules of neuroendocrine cells and plays an important role in the release of neurotransmitters.
  • Synaptophysin can interact with VAMP2 and dissociate from VAMP2 before it forms a SNARE complex with SNAP25 and Syntaxin, thus regulating the formation of the SNARE complex, resulting in reduced neurotransmitter release under high-frequency stimulation. Abnormal regulation of synaptophysin can lead to abnormal neuronal exocytosis and muscle contraction impairment.
  • polypeptide fragments derived from Synaptophysin can interfere with the interaction between Synaptophysin and VAMP2, and thus inhibit the formation of SNARE complexes and the release of neurotransmitters (preferably acetylcholine), thereby inhibiting muscle contraction, and therefore can be used for treatment or The care of conditions, disorders, or diseases caused by muscle contraction, and the treatment or care of the skin.
  • neurotransmitters preferably acetylcholine
  • the present application provides a peptide represented by formula (I), or a stereoisomer thereof, or a mixture of stereoisomers thereof, or a cosmetically acceptable salt thereof, or a pharmaceutically acceptable salt thereof ,
  • X 1 is selected from: -Val-, -Ser-, -Trp-, -Met-, -Leu-, -Glu-, -Ala-, -Pro-, -Tyr- or -Thr-;
  • X 2 is selected from: -Lys-, -Arg-, -His-, -Met-, -Ala-, -Leu-, -Asn-, -Trp-, -Thr-, -Pro-, -Gly-, - Phe-, -Asp-, -Ser- or -Cys-;
  • X 3 is selected from: -Val-, -Gly-, -Leu-, -Pro-, -Met-, -Ala-, -Gln-, -Asp-, -Phe-, -Ser-, -Cys- or - Lys-;
  • X 4 is selected from: -Gly-, -Leu-, -Pro-, -Ala-, -Phe-, -Val-, -Trp-, -Asn- or -Glu-;
  • X 5 is selected from: -Phe-, -Trp-, -Tyr- or bond;
  • X 6 is selected from: -Gln-, -Leu-, -Ala-, -Val-, -Phe-, -Arg-, -Tyr-, -Gly-, or -Asn-;
  • X 7 is selected from: -Lys-, -Arg-, -His- or bond;
  • X 8 is selected from: -Trp-, -Ala-, -Asp-, -Lys-, -Gly-, -Tyr-, -Met- or -Arg-;
  • R 1 is selected from: H or R 3 -CO-, where R 3 is selected from: substituted or unsubstituted alkyl, substituted or unsubstituted alkenyl;
  • R 2 is selected from: -NR 4 R 5 or -OR 4 , wherein each R 4 and R 5 are independently selected from: H, substituted or unsubstituted alkyl, substituted or unsubstituted alkenyl;
  • the alkyl group refers to having 1-24 carbon atoms (optional 1-16 carbon atoms; optional 1-14 carbon atoms; optional 1-12 carbon atoms; optional 1, 2 , 3, 4, 5, or 6 carbon atoms) saturated aliphatic linear or branched alkyl group; optionally selected from: methyl, ethyl, isopropyl, isobutyl, tert-butyl, Pentyl, hexyl, heptyl, octyl, decyl, dodecyl, tetradecyl, hexadecyl, octadecyl, 2-ethylhexyl, 2-methylbutyl, or 5- methylhexyl;
  • the alkenyl group refers to having 2-24 carbon atoms (optional 2-16 carbon atoms; optional 2-14 carbon atoms; optional 2-12 carbon atoms; optional 2, 3 , 4, 5, or 6 carbon atoms) linear or branched alkenyl; the alkenyl has one or more carbon-carbon double bonds, optionally with 1, 2, or 3 conjugated or non-conjugated a carbon-carbon double bond; the alkenyl group is bonded to the rest of the molecule through a single bond; optionally selected from: vinyl, oleyl, or linoleyl;
  • R 1 is selected from: H, acetyl, tert-butyryl, caproyl, 2-methylcaproyl, octanoyl, decanoyl, lauroyl, myristoyl, palmitoyl, stearoyl, oleoyl or linoleoyl;
  • R 1 is selected from H, acetyl, lauroyl, myristoyl or palmitoyl;
  • R1 is H or acetyl.
  • R 4 and R 5 are independently selected from: H, methyl, ethyl, hexyl, dodecyl or hexadecyl;
  • R 4 is H and R 5 is selected from: H, methyl, ethyl, hexyl, dodecyl or hexadecyl;
  • R2 is -OH or -NH2 .
  • X 5 is a key and X 7 is a key.
  • peptide represented by formula (I), or a stereoisomer thereof, or a mixture of stereoisomers thereof, or a cosmetically acceptable salt thereof, or a pharmaceutically acceptable salt thereof is selected from the following peptides (1)-(48):
  • the peptide, or a stereoisomer thereof, or a mixture of stereoisomers thereof, or a cosmetically acceptable salt thereof, or a pharmaceutically acceptable salt thereof is selected from the group consisting of Ac-Val-Lys-Val -Leu-Gln-Trp-NH 2 , Ac-Ser-Met-Gly-Ala-Leu-Ala-NH 2 or Ac-Trp-Ala-Ala-Pro-Phe-Leu-Arg-Ala-NH 2 .
  • the peptide represented by formula (I) of the present application may exist as a stereoisomer or a mixture of stereoisomers; for example, the amino acids contained therein may have L-, D- configurations, or may be independently of each other. Racemic. It is therefore possible to obtain isomeric mixtures as well as racemic or diastereomeric mixtures, or pure diastereomers or enantiomers, depending on the number of asymmetric carbons and what isomeric mixtures are present. Isomers or isomeric mixtures.
  • the preferred structure of the peptide represented by formula (I) of the present application is a pure isomer, that is, an enantiomer or a diastereomer.
  • -Phe- is mentioned in this application, it should be understood that -Phe- is selected from -L-Phe-, -D-Phe-, or a mixture of the two, whether racemic or non-racemic.
  • the preparation methods described in this document enable one of ordinary skill in the art to obtain each stereoisomer of the peptides of the present application by selecting amino acids with the correct configuration.
  • the present application also includes all suitable isotopic variants of the peptide represented by formula (I).
  • Isotopic variants of the peptides of the present application are here understood to mean compounds in which at least one atom within the peptide of the present application is replaced by another atom of the same atomic number, but with a different atomic mass of said other atom Atomic mass commonly or predominantly found in nature.
  • isotopes that may be incorporated into the peptides of the present application are: those of hydrogen, carbon, nitrogen, oxygen or sulfur, such as 2 H (deuterium), 3 H (tritium), 13 C, 14 C, 15 N, 17 O , 18 O, 33 S, 34 S, 35 S or 36 S.
  • isotopic variants of the peptides of the present application may be advantageous, for example, to examine the mechanism of action or the distribution of the active compound in vivo; due to the relative simplicity of their preparation properties and detectability, especially compounds labeled with 3 H or 14 C isotopes are suitable for this purpose.
  • isotopes e.g., deuterium
  • Such modifications of peptides may also constitute preferred embodiments of the present application.
  • the present invention can be prepared by methods known to the person skilled in the art, for example by the methods described further below and described in the examples, by using the respective reagents and/or corresponding isotopic modifications of the starting substances. Isotopic variants of the applied peptide.
  • the present application also includes prodrugs of the peptides of the present application.
  • prodrug is used herein to mean compounds which may themselves be biologically active or inactive, but which, during their residence time in the body, react (e.g., metabolize or hydrolyze) to produce the product of the present invention. of peptides.
  • compositions refers to a salt approved for use in animals, and more specifically in humans, including metal salts of the peptides represented by formula (I),
  • the metals include, but are not limited to: lithium, sodium, potassium, calcium, magnesium, manganese, copper, zinc or aluminum, etc.; including salts formed by the peptide represented by formula (I) and organic bases, and the organic bases include : Ethylamine, diethylamine, ethylenediamine, ethanolamine, diethanolamine, arginine, lysine, histidine or piperazine; including salts formed by the peptide represented by formula (I) and inorganic or organic acids , 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, trisaccharide, Fluoroace
  • the synthesis of the peptide represented by formula (I) of the present application, or its stereoisomer or its cosmetically acceptable salt, or its pharmaceutically acceptable salt can be carried out according to conventional methods known in the art, For example, solid-phase synthesis, liquid-phase synthesis, or a combination of solid-phase and liquid-phase methods, biotechnological methods aimed at producing the desired sequence, or control of proteins of animal, fungal, or plant origin can also be used. Prepared by hydrolysis.
  • a method for obtaining the peptide represented by formula (I) includes the following steps:
  • the C-terminus is bound to a solid support and the method is performed on a solid phase and includes combining an amino acid having a protected N-terminus and a free C-terminus with an amino acid having a free N-terminus and with a Coupling of amino acids at the C-terminus bound to the polymeric carrier; elimination of the group protecting the N-terminus; and repeating this sequence as many times as necessary to thereby obtain a peptide of the desired length, followed by cleavage of the desired length from the original polymeric carrier Synthetic peptides.
  • the functional groups of the side chains of these amino acids remain fully protected with temporary or permanent protecting groups throughout the synthesis and can be deprotected simultaneously or orthogonally with the process of cleaving the peptide from the polymeric support.
  • solid phase synthesis can be performed by a convergent strategy in which dipeptides or tripeptides are coupled to a polymeric support or to a dipeptide or amino acid previously bound to a polymeric support.
  • the functional groups of the termini can then be modified by deprotecting the N- and C-termini and/or cleaving the peptide from the polymer support in an undetermined order using standard conditions and methods known in the art.
  • Optional modifications to the N-terminus and C-terminus may be made to the peptide of formula (I) bound to the polymer support or after the peptide has been cleaved from the polymer support. Selected modifications.
  • Another aspect of the present application provides a cosmetic or pharmaceutical composition, including an effective amount of the peptide represented by the above formula (I), or a stereoisomer thereof, or a mixture of stereoisomers, or a cosmetic composition thereof. an acceptable salt, or a pharmaceutically acceptable salt thereof, and at least one excipient and optionally a cosmetically or pharmaceutically acceptable adjuvant.
  • the adjuvant is selected from: collagen synthesis stimulators, agents that regulate PGC-1 ⁇ synthesis, agents that regulate the activity of PPAR ⁇ , agents that increase or decrease the triglyceride content of adipocytes, and stimulate or delay adipocyte differentiation.
  • the preparation of the cosmetic or medicinal composition is selected from: cream, oil, milk, balm, foam, lotion, gel, liniment, serum, soap, shampoo, conditioner, serum , ointment, mousse, pomade, powder, rod, pen, spray, aerosol, capsule, tablet, granule, chewing gum, solution, suspension, emulsion, syrup, elixir, polysaccharide Film, jelly or gelatin;
  • the capsules include: soft capsules, hard capsules, optionally gelatin capsules; the tablets include: sugar-coated tablets.
  • the peptides of the present application have variable solubility in water depending on the nature of their sequence or any possible modifications in the N-terminus and/or C-terminus.
  • the peptides of the present application may therefore be incorporated into compositions via aqueous solutions, and those that are insoluble in water may be soluble in conventional cosmetically or pharmaceutically acceptable solvents such as and without limitation ethanol, propanol, isopropanol , propylene glycol, glycerin, butylene glycol or polyethylene glycol or any combination thereof.
  • the cosmetically or pharmaceutically effective amounts of the peptides of the present application to be administered, as well as their dosage, will depend on many factors, including age, condition of the patient, severity of condition or disease, route and frequency of administration, and the nature of the peptide to be used. specific nature.
  • Cosmetically or pharmaceutically effective amount means an amount of one or more peptides of the present application that is non-toxic but sufficient to provide the desired effect.
  • the peptides of the present application are used in the cosmetic or pharmaceutical compositions of the present application at a cosmetically or pharmaceutically effective concentration to obtain the desired effect; in a preferred form, at 0.00000001% relative to the total weight of the composition (by weight) and 20% (by weight), preferably between 0.000001% (by weight) and 15% (by weight), more preferably between 0.0001% (by weight) and 10% ( between 0.0001% by weight and 5% by weight.
  • Another aspect of the present application provides a cosmetically or pharmaceutically acceptable delivery system or sustained release system in order to achieve better penetration of the active ingredient and/or improve its pharmacokinetic and pharmacodynamic properties, It contains an effective amount of the peptide represented by the above formula (I), or a stereoisomer thereof, or a mixture of stereoisomers, or a cosmetically acceptable salt thereof, or a pharmaceutically acceptable salt thereof, or the above cosmetic or medicinal compositions.
  • delivery system refers to a diluent, adjuvant, excipient or carrier administered with the peptides of the present application, which are selected from: water, oil or surfactant, including petroleum sources, animal sources, plant sources, or Those of synthetic origin, such as and not limited to peanut oil, soybean oil, mineral oil, sesame oil, castor oil, polysorbates, sorbitan esters, ether sulfates, sulfates, betaines, glucosides, maltosides, fats alcohol, nonoxynol ether, poloxamer, polyoxyethylene, polyethylene glycol, dextrose, glycerin, digitonin and the like.
  • Those of ordinary skill in the art are aware of the diluents that can be used in the different delivery systems in which the peptides of the present application can be administered.
  • sustained release is used in its conventional sense and refers to a delivery system that provides a gradual release of the compound over a period of time, and preferably, but not necessarily, a relatively constant level of release of the compound throughout the period of time.
  • Examples of delivery systems or sustained release systems are liposomes, oleosomes, nonionic surfactant liposomes, ethosomes, millimeter capsules, microcapsules, nanocapsules, nanostructured lipid carriers, sponges substances, cyclodextrins, lipid vesicles, micelles, millimeter spheres, microspheres, nanospheres, lipid spheres, microemulsions, nanoemulsions, millimeter particles, microparticles or nanoparticles.
  • Preferred delivery systems or sustained release systems are liposomes and microemulsions, more preferably water-in-oil microemulsions having an internal structure of reverse micelles.
  • Sustained release systems may be prepared by methods known in the art and may be administered, for example, by topical or transdermal administration, including adhesive patches, non-adhesive patches, occlusive patches, and microelectronic patch; or by systemic administration such as, and without limitation, oral or parenteral routes, including nasal, rectal, subcutaneous implantation or injection, or direct implantation or injection into a specific body site, and preferably should Relatively constant amounts of these peptides of the present application are released.
  • topical or transdermal administration including adhesive patches, non-adhesive patches, occlusive patches, and microelectronic patch
  • systemic administration such as, and without limitation, oral or parenteral routes, including nasal, rectal, subcutaneous implantation or injection, or direct implantation or injection into a specific body site, and preferably should Relatively constant amounts of these peptides of the present application are released.
  • the amount of peptide included in the sustained release system will depend, for example, on the site to which the composition is to be administered, the release kinetics and duration of the peptides of the present application, and the condition, disorder and/or condition to be treated and/or cared for. Nature of disease.
  • Another aspect of the present application provides a peptide represented by the above formula (I), or a stereoisomer thereof, or a mixture of stereoisomers, or a cosmetically acceptable salt thereof, or a pharmaceutically acceptable salt thereof.
  • the disorder or disease is dystonia, specifically, the dystonia is focal dystonia; dystonias include, but are not limited to, facial spasm, torsional dystonia, cervical dystonia or strabismus. Cervical dystonia, laryngeal dystonia or spasmodic dysphonia, oromandibular dystonia, limb dystonia such as writer's cramp or musician's cramp or foot dystonia, nocturnal bruxism, hemifacial spasm, tics and/or strabismus; segments dystonia, Major syndrome, multifocal dystonia, hemidystonia, dopamine-responsive dystonia, and Segawa dystonia.
  • Another aspect of the present application provides a peptide represented by the above formula (I), or a stereoisomer thereof, or a mixture of stereoisomers, or a cosmetically acceptable salt thereof, or a pharmaceutically acceptable salt thereof.
  • Use in a composition, wherein the treatment, prevention or repair of skin aging or photoaging is the reduction, prevention or treatment of facial wrinkles.
  • the term “skin” is understood to mean the layers constituting it, from the uppermost layer or stratum corneum to the lowermost layer or subcutaneous tissue, both ends included. These layers are composed of different types of cells, such as keratinocytes, fibroblasts, melanocytes, and/or adipocytes. In this application, the term “skin” includes the scalp.
  • treatment refers to administration of a peptide according to the present application to reduce or eliminate a disease or condition, or to reduce or eliminate one or more symptoms associated with such disease or condition.
  • treatment also encompasses the ability to reduce or eliminate the physiological consequences of the disease or condition.
  • care includes the prevention of disease and/or disorder.
  • prevention refers to the ability of the peptides of the present application to prevent, delay, or hinder the occurrence or progression of a disease or condition before it occurs.
  • aging refers to the changes that the skin undergoes as we age (natural aging), or through exposure to sunlight (photoaging) or exposure to environmental pollutants such as chemical dirt or pollutants, tobacco smoke, etc. Changes, and includes all changes that are externally visible and/or perceptible by touch, such as and without limitation: the development of discontinuities in the skin (such as wrinkles, fine lines, expression lines, stretch lines, streaks, grooves, lines, unevenness or roughness, increased pore size, loss of moisture, loss of elasticity, loss of firmness, loss of smoothness, loss of recovery from deformation, loss of resilience), skin sagging (such as sagging cheeks, bags under the eyes, or Double chin, etc.), changes in skin color (such as scars, redness, eye bags, or hyperpigmentation areas such as age spots or freckles, etc.), abnormal differentiation, hyperkeratinization, elastosis, keratosis, alopecia, orange Dermoid skin, loss of collagen structure, and other histological changes in
  • photoaging refers to the premature aging of the skin due to long-term exposure of the skin to ultraviolet radiation, which exhibits the same physiological characteristics as natural aging, such as and not limited to: sagging, sagging, color changes or hypopigmentation. Regular, abnormal and/or hyperkeratinization.
  • Lys means NH 2 -CH(CH 2 CH 2 CH 2 CH 2 NH 2 )-COOH
  • Lys- means NH 2 -CH(CH 2 CH 2 CH 2 CH 2 NH 2 )-CO-
  • -Lys represents -NH-CH(CH 2 CH 2 CH 2 CH 2 NH 2 )-COOH
  • -Lys- represents -NH-CH(CH 2 CH 2 CH 2 CH 2 NH 2 )-CO-.
  • Ac- is used in this application to represent acetyl (CH 3 -CO-); Ser: serine; Gly: glycine; Ala: alanine; Lys: lysine; Trp: tryptophan; Val: Valine; Leu: leucine; Gln: glutamine; Met: methionine; His: histidine; Arg: arginine; Pro: proline; Phe: phenylalanine; Tyr: Tyrosine.
  • the polypeptide of the present application is artificially designed and is easy to synthesize. Through interaction with synaptic vesicle-associated membrane protein (VAMP2), it regulates the formation of SNARE complex, thereby interfering with the release of synaptic neurotransmitters, inhibiting muscle contraction, and improving It can reduce wrinkles on the surface of the skin and reduce the depth of wrinkles to achieve anti-wrinkle effects. It can be used in the field of cosmetics or medicine to improve skin aging or photoaging, and to treat or care for conditions, disorders or diseases caused by muscle contraction.
  • VAMP2 synaptic vesicle-associated membrane protein
  • Figure 1 is the mass spectrum of peptide (4)Ac-Val-Lys-Val-Leu-Gln-Trp-NH 2 (molecular formula C 40 H 64 N 10 O 8 ), the mass charge of [M+H] + quasi-molecular ion peak The ratio (m/z) is 813.5879, and the molecular weight measured by mass spectrometry is 812.59;
  • Figure 2 is the mass spectrum of peptide (8)Ac-Ser-Met-Gly-Ala-Leu-Ala-NH 2 (molecular formula C 24 H 43 N 7 O 8 S), and the mass of the [M+Na] + addition ion peak
  • the charge ratio (m/z) is 612.3438, and the molecular weight measured by mass spectrometry is 589.34;
  • Figure 3 is the mass spectrum of peptide (32)Ac-Trp-Ala-Ala-Pro-Phe-Leu-Arg-Ala-NH 2 (molecular formula C 48 H 69 N 13 O 9 ), [M+H] + quasi-molecular ion
  • the mass-to-charge ratio (m/z) of the peak is 972.6496, and the molecular weight measured by mass spectrometry is 971.65;
  • Figure 4 is a graph showing the results of the muscle contraction inhibition experiment on the test sample.
  • the core concept of this application is to obtain a biomimetic peptide based on Synaptophysin. These peptides can interfere with the interaction between Synaptophysin and VAMP2, thereby inhibiting the formation of SNARE complexes and neurotransmitter release, thereby inhibiting muscle contraction.
  • the polypeptide has the following general formula: R 1 -X 1 -X 2 -X 3 -X 4 -X 5 -X 6 -X 7 -X 8 -R 2 ; the peptide of the present application, its stereoisomers, and stereoisomers Mixtures of isomers, or cosmetically acceptable salts thereof, or pharmaceutically acceptable salts thereof, or cosmetic compositions or pharmaceutical compositions thereof, and their preparations for improving skin aging or photoaging, treating or use in cosmetic or pharmaceutical compositions for the care of conditions, disorders or diseases caused by muscle contraction.
  • Amide Resin a starting resin for polypeptide synthesis (cross-linking degree 1%, substitution degree 1.72mmol/g, particle size 100-200 mesh);
  • Fmoc-Linker 4-[(2,4-dimethoxybenzene (Fmoc-amino)methyl]phenoxyacetic acid; Ac 2 O: acetic anhydride; DMF: N,N-dimethylformamide; DIPEA: diisopropylethylamine; DIC: diisopropylcarbodiamine Imine; piperidine: piperidine; HOBt: 1-hydroxybenzotriazole; TFA: trifluoroacetic acid; TIS: triisopropylsilane; EDT: 1,2-ethanedithiol; Ser: serine; Gly: Glycine; Ala: alanine; Lys: lysine; Trp: tryptophan; Val: valine; Leu: leucine; Gln: glutamine; Met: methionine;
  • VAMP2 In order to further evaluate the binding ability of the polypeptide to VAMP2, VAMP2 before participating in the formation of the SNARE complex was selected for study. Molecular docking and molecular dynamics were used to calculate the affinity of the above peptides with VAMP2, and then the best candidate peptides were screened and considered for further activity verification. The best candidate peptides are shown in Table 2 below.
  • Fmoc-Linker-Amide Resin was removed from Fmoc twice with 20% piperidine/DMF, 10 minutes each time. Samples were taken for K inspection and the color was dark blue. Wash the resin 7 times with DMF and remove the solvent.
  • the N-terminal Fmoc group was deprotected and 7.3g of activated Fmoc-Gln(Trt)-OH was coupled to the peptidyl resin using DMF as solvent in the presence of 1.8g HOBt and 1.65g DIC. on, and continue to react for 2 hours. The resins are then washed and deprotection of the Fmoc group is repeated for coupling of the next amino acid.
  • 4.2 g of Fmoc-Leu-OH; 4.0 g of Fmoc-Val-OH; 5.6 g of Fmoc-Val-OH were sequentially coupled using DMF as solvent in the presence of 1.8 g of HOBt and 1.65 g of DIC.
  • Fmoc-Lys(Boc)-OH followed by 4.0 g of Fmoc-Val-OH; after the reaction was complete, the resin was washed and the solvent was removed.
  • Fmoc-Linker-Amide Resin was removed from Fmoc twice with 20% piperidine/DMF, 10 minutes each time. Samples were taken for K inspection and the color was dark blue. Wash the resin 8 times with DMF and remove the solvent.
  • the N-terminal Fmoc group was deprotected, and 4.2g of activated Fmoc-Leu-OH was coupled to the peptidyl resin using DMF as a solvent in the presence of 1.8g HOBt and 1.65g DIC. Continue Reaction 2h. The resins are then washed and deprotection of the Fmoc group is repeated for coupling of the next amino acid.
  • Fmoc-Linker-Amide Resin was removed from Fmoc twice with 20% piperidine/DMF, 10 minutes each time. Samples were taken for K inspection and the color was dark blue. Wash the resin 7 times with DMF and remove the solvent.
  • the N-terminal Fmoc group was deprotected and 7.6g of activated Fmoc-Arg(Pbf)-OH was coupled to the peptidyl resin using DMF as solvent in the presence of 1.8g HOBt and 1.65g DIC. on, and continue to react for 2 hours. The resins are then washed and deprotection of the Fmoc group is repeated for coupling of the next amino acid.
  • HW-400S constant temperature smooth muscle tank BL-420E biological function experimental system, tension transducer, and other instruments are conventional experimental instruments.
  • Acetylcholine chloride (Shanghai Yuanye), chloral hydrate (Shanghai McLean)
  • Acetyl octapeptide-3 (Ac-Glu-Glu-Met-Gln-Arg-Arg-Ala-Asp-NH 2 ), the test concentrations are 25ppm, 50ppm, 100ppm, and 200ppm respectively;
  • Acetyl hexapeptide-30 (Ac-Glu-Asp-Tyr-Tyr-Arg-Leu-NH 2 ), the test concentrations are 25ppm, 50ppm, 100ppm, and 200ppm respectively;
  • test concentrations are 25ppm, 50ppm, 100ppm, and 200ppm respectively;
  • test concentrations are 25ppm, 50ppm, 100ppm, and 200ppm respectively;
  • test concentrations were 100ppm, 200ppm, 400ppm, and 800ppm respectively.
  • the experimental rats were fasted and water-free for 12 hours before the experiment. Inject 10% chloral hydrate intraperitoneally at a dose of 0.1mL/25g. After the rat is anesthetized, the abdominal cavity is cut open to expose the enlarged cecum. A section of ileum is taken out from the ileocecal intestine and washed in benchtop solution that has been pre-oxygenated for 10 minutes. Cut several 4cm long sections of ileum and incubate them in 4°C benchtop liquid for later use; partially ligate and tie them with cotton thread at both ends of the ileum to form a ring;
  • Ileal smooth muscle contraction inhibition rate (contractile force after acetylcholine administration - contractile force after test sample administration)/contractile force after acetylcholine administration ⁇ 100%
  • the inhibition rate at 50 ppm was 32.34%, the inhibition rate at 100 ppm was 47.49%, and the inhibition rate at 200 ppm was 58.67%; however, giving peptide (4) and peptide (8) has a significant inhibitory effect on the high signal peak produced by acetylcholine at 25ppm, and as the concentration increases, the inhibitory effect increases; peptide (4) produces an inhibitory effect at 25ppm, with an inhibition rate of 49.78%, and at 50ppm
  • the inhibition rate is 62.40%, the inhibition rate at 100ppm is 66.72%, and the inhibition rate at 200ppm concentration is as high as 76.88%; the inhibition rate of peptide (8) is 31.99% at 25ppm, 34.35% at 50ppm, and 34.35% at 100ppm.
  • the inhibition rate was 56.42%, and the inhibition rate was as high as 67.96% at a concentration of 200 ppm; administration of peptide (32) at 800 ppm produced an inhibitory effect on muscle contraction
  • the peptide of the present application has an obvious effect of inhibiting muscle contraction, and is stronger than the prior art polypeptides; its inhibitory efficiency is peptide (4)>peptide (8)>acetyl octapeptide-3> Acetyl hexapeptide-30.
  • Acetylcholine as a neurotransmitter, can directly stimulate acetylcholine receptors on smooth muscles, triggering a signal cascade reaction and causing smooth muscles to contract.
  • muscle contraction can be inhibited, conditions, disorders or diseases caused by muscle contraction can be treated or treated, skin aging or photoaging can be treated, prevented or repaired, facial wrinkles can be reduced, prevented or treated .
  • the peptide of the present application interferes with the interaction between Synaptophysin and VAMP2, affects the formation of SNARE complexes between VAMP2, SNAP25 and Syntaxin, thereby inhibiting the release of acetylcholine, thereby inhibiting muscle contraction.
  • This experiment further clarifies the technical effect of the peptide of the present application on inhibiting muscle contraction by detecting the contraction signal of ileal smooth muscle and comparing the inhibition rate of ileal smooth muscle contraction of each sample.
  • Preparation method Prepare peptide (4) into an aqueous solution of 0.02 mg/mL; heat cetearyl alcohol (and) cetearyl glucoside, jojoba oil, mineral oil, and isopropyl palmitate to 85°C , stir evenly; phase A is obtained; dissolve glycerin, allantoin, polyacrylamide (and) C13-14 isoparaffin (and) laureth-7 with water, and heat to 85°C to obtain phase B; dissolve A Quickly add phase B to phase B, homogenize at constant temperature for 3-5 minutes, and cool; cool to below 60°C, add preservatives, stir evenly, cool to below 45°C, add polypeptide solution and essence, and you have it.
  • Preparation method Add the prescribed amount of sodium hyaluronate to water, stir to mix evenly, then heat to 80-85°C, keep warm and stir to disperse evenly; when the temperature drops below 40°C, add glycerin, aloe vera gel, peptide (8 ), vitamin C, caprylyl glycol and 1,2-hexanediol, stir evenly; use triethanolamine solution to adjust the pH value of the solution to about 5.5.

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Abstract

本申请提供了一种多肽及其美容组合物或药用组合物和用途，所述多肽具有以下通式：R 1-X 1-X 2-X 3-X 4-X 5-X 6-X 7-X 8-R 2；本申请的肽、其立体异构体、立体异构体的混合物、或其美容上可接受的盐、或其药学上可接受的盐、或它们的美容组合物或药用组合物、以及它们在制备用于改善皮肤老化或光老化，治疗或护理由肌肉收缩导致的病况、失调或疾病的美容组合物或药物组合物中的用途。

Description

一种多肽及其美容组合物或药用组合物和用途 技术领域
本申请涉及医药和化妆品技术领域，特别是一种多肽及其美容组合物或药用组合物和用途。
背景技术
人体皮肤衰老最明显的信号是脸部皱纹的产生。随着时间推移，受年龄增长、外界环境和遗传因子等多方面因素的影响，机体的神经活动过度，刺激肌肉纤维的神经递质不受控制和过度释放，从而向内拖动皮肤的肌肉张力，使皮肤表面一级线的加深从而形成皱纹。在细胞水平上，位于张力线上的合成胞外基质和胶原的成纤维细胞可在肌肉收缩的作用下发挥与横纹肌有关的特定收缩性质。神经与横纹肌之间的接合点构成神经肌肉板，在其上游是运动神经元传入神经通路，已知面部的皮肤肌肉处于运动神经传入的控制之下，通过从神经突触囊泡中释放乙酰胆碱，作用于突触后膜的乙酰胆碱受体使肌肉收缩，导致皱纹形成。
囊泡需先与突触前膜融合才能完成胞吐过程，释放乙酰胆碱。膜融合由N-乙基马来酰亚胺敏感因子附着蛋白受体(SNARE)复合物介导。SNARE复合物为四螺旋束结构，由位于囊泡膜上的突触小泡缔合性膜蛋白(VAMP2)、位于突触前膜的突触相关蛋白(SNAP25)和突触融合蛋白(Syntaxin)构成。SNARE通过在聚合时构象发生变化并释放能量，促使囊泡膜和突触前膜融合以及神经递质的释放，从而参与神经元的胞吐作用和肌肉收缩活动。Synaptophysin(突触体素)是一种突触囊泡跨膜糖蛋白，存在于神经元的突触前小泡和神经内分泌细胞的神经分泌颗粒中，在神经递质释放过程中发挥重要作用。Synaptophysin可以与VAMP2相互作用，在VAMP2与SNAP25和Syntaxin形成SNARE复合物之前与VAMP2分离，从而调节SNARE复合物的形成，导致高频刺激下神经递质的释放减少。Synaptophysin的异常调节会导致异常的神经元胞吐作用和肌肉收缩障碍。
目前，人们以SNARE蛋白复合物为靶点，已经发现了几种用于减少和/或消除表情皱纹的方法。例如，通过局部注射肉毒杆菌毒素A用于除皱、瘦脸、消除眉间纹和鱼尾纹等，但存在引发免疫反应而导致治疗作用丧失的风险；专利WO0064932、EP1180524B1和EP2123673B1中源自SNAP-25氨基和羧基结构域的肽；WO2019166347中衍生自Munc18的肽。然而，目前尚缺少能够以类似于肉毒杆菌毒素的有效性，但无风险的方式抑制肌肉收缩的化合物。因此，仍然需要寻找其他作用机制的化合物来抑制神经元胞吐作用，干扰突触神经递质释放，从而满足日益增长的治疗或护理皮肤的需求。
发明内容
本申请的发明人研究发现，源自Synaptophysin的多肽片段可以干扰Synaptophysin与VAMP2相互作用，并因此抑制SNARE复合物的形成和神经递质释放(优选乙酰胆碱)，从而抑制肌肉收缩，因此可用于治疗或护理由肌肉收缩导致的病况、失调或疾病，以及治疗或护理皮肤。
鉴于此，本申请提供一种式(I)所示的肽，或其立体异构体、或其立体异构体的混合物，或其 美容上可接受的盐、或其药学上可接受的盐，
R 1-X 1-X 2-X 3-X 4-X 5-X 6-X 7-X 8-R 2   (I)
式(I)中，
X 1选自：-Val-、-Ser-、-Trp-、-Met-、-Leu-、-Glu-、-Ala-、-Pro-、-Tyr-或-Thr-；
X 2选自：-Lys-、-Arg-、-His-、-Met-、-Ala-、-Leu-、-Asn-、-Trp-、-Thr-、-Pro-、-Gly-、-Phe-、-Asp-、-Ser-或-Cys-；
X 3选自：-Val-、-Gly-、-Leu-、-Pro-、-Met-、-Ala-、-Gln-、-Asp-、-Phe-、-Ser-、-Cys-或-Lys-；
X 4选自：-Gly-、-Leu-、-Pro-、-Ala-、-Phe-、-Val-、-Trp-、-Asn-或-Glu-；
X 5选自：-Phe-、-Trp-、-Tyr-或键；
X 6选自：-Gln-、-Leu-、-Ala-、-Val-、-Phe-、-Arg-、-Tyr-、-Gly-、或-Asn-；
X 7选自：-Lys-、-Arg-、-His-或键；
X 8选自：-Trp-、-Ala-、-Asp-、-Lys-、-Gly-、-Tyr-、-Met-或-Arg-；
R 1选自：H或R 3-CO-，其中R 3选自：取代的或未取代的烷基、取代的或未取代的烯基；
R 2选自：-NR 4R 5或-OR 4，其中各个R 4和R 5彼此独立地选自：H、取代的或未取代的烷基、取代的或未取代的烯基；
所述烷基是指具有1-24个碳原子(可选具有1-16个碳原子；可选具有1-14个碳原子；可选具有1-12个碳原子；可选具有1、2、3、4、5、或6个的碳原子)的饱和脂肪族直链或支链的烷基；可选选自：甲基、乙基、异丙基、异丁基、叔丁基、戊基、己基、庚基、辛基、癸基、十二烷基、十四烷基、十六烷基、十八烷基、2-乙基己基、2-甲基丁基、或5-甲基己基；
所述烯基是指具有2-24个碳原子(可选具有2-16个碳原子；可选具有2-14个碳原子；可选具有2-12个碳原子；可选具有2、3、4、5、或6个碳原子)的直链或支链烯基；所述烯基具有一个或多个碳-碳双键，可选具有1、2或3个共轭或非共轭的碳-碳双键；所述烯基是通过一个单键而结合至分子的其余部分；可选选自：乙烯基、油烯基、或亚油烯基；
可选地，所述“取代的烷基”、“取代的烯基”中的取代基选自C 1-C 4烷基；羟基；C 1-C 4烷氧基；氨基；C 1-C 4氨基烷基；C 1-C 4羰氧基；C 1-C 4氧基羰基；卤素(如氟、氯、溴、以及碘)；氰基；硝基；叠氮化物；C 1-C 4烷基磺酰基；硫醇；C 1-C 4烷硫基；C 6-C 30芳氧基如苯氧基；-NR b(C＝NR b)NR bR c，其中R b和R c是独立地选自：H、C 1-C 4烷基、C 2-C 4烯基、C 2-C 4炔基、C 3-C 10环烷基、C 6-C 18芳基、C 7-C 17芳烷基、具有三至十元的杂环基、或氨基的保护基。
进一步地，R 1选自：H、乙酰基、叔-丁酰基、己酰基、2-甲基己酰基、辛酰基、癸酰基、月桂酰基、肉豆蔻酰基、棕榈酰基、硬脂酰基、油酰基或亚油酰基；
其中，R 1选自H、乙酰基、月桂酰基、肉豆蔻酰基或棕榈酰基；
可选地，R 1是H或乙酰基。
进一步地，R 4、R 5彼此独立地选自：H、甲基、乙基、己基、十二烷基或十六烷基；
其中，R 4是H并且R 5选自：H、甲基、乙基、己基、十二烷基或十六烷基；
可选地，R 2是-OH或-NH 2。
进一步地，X 5是键，X 7是键。
进一步地，式(I)所示的肽，或其立体异构体、或其立体异构体的混合物，或其美容上可接受的盐、或其药学上可接受的盐，选自下列肽(1)-(48)：
(1)H-Val-Lys-Val-Leu-Gln-Trp-OH；
(2)H-Val-Lys-Val-Leu-Gln-Trp-NH 2；
(3)Ac-Val-Lys-Val-Leu-Gln-Trp-OH；
(4)Ac-Val-Lys-Val-Leu-Gln-Trp-NH 2；
(5)H-Ser-Met-Gly-Ala-Leu-Ala-OH；
(6)H-Ser-Met-Gly-Ala-Leu-Ala-NH 2；
(7)Ac-Ser-Met-Gly-Ala-Leu-Ala-OH；
(8)Ac-Ser-Met-Gly-Ala-Leu-Ala-NH 2；
(9)Ac-Val-His-Pro-Leu-Gln-Trp-OH；
(10)Ac-Val-His-Pro-Leu-Gln-Trp-NH 2；
(11)Ac-Ser-Arg-Leu-Pro-Leu-Trp-OH；
(12)Ac-Ser-Arg-Leu-Pro-Leu-Trp-NH 2；
(13)Ac-Trp-Lys-Val-Pro-Gln-Ala-OH；
(14)Ac-Trp-Lys-Val-Pro-Gln-Ala-NH 2；
(15)Ac-Val-Arg-Leu-Gly-Phe-Gln-His-Trp-OH；
(16)Ac-Val-Arg-Leu-Gly-Phe-Gln-His-Trp-NH 2；
(17)Ac-Val-His-Pro-Leu-Phe-Gln-His-Trp-OH；
(18)Ac-Val-His-Pro-Leu-Phe-Gln-His-Trp-NH 2；
(19)Ac-Val-Ala-Val-Leu-Phe-Gln-Arg-Trp-OH；
(20)Ac-Val-Ala-Val-Leu-Phe-Gln-Arg-Trp-NH 2；
(21)Ac-Val-Met-Gly-Pro-Trp-Leu-Lys-Ala-OH；
(22)Ac-Val-Met-Gly-Pro-Trp-Leu-Lys-Ala-NH 2；
(23)Ac-Val-Met-Ala-Pro-Tyr-Leu-Lys-Ala-OH；
(24)Ac-Val-Met-Ala-Pro-Tyr-Leu-Lys-Ala-NH 2；
(25)Ac-Ser-Ala-Val-Pro-Trp-Leu-Arg-Trp-OH；
(26)Ac-Ser-Ala-Val-Pro-Trp-Leu-Arg-Trp-NH 2；
(27)Ac-Ser-Met-Ala-Gly-Trp-Gln-His-Ala-OH；
(28)Ac-Ser-Met-Ala-Gly-Trp-Gln-His-Ala-NH 2；
(29)H-Trp-Ala-Ala-Pro-Phe-Leu-Arg-Ala-OH；
(30)H-Trp-Ala-Ala-Pro-Phe-Leu-Arg-Ala-NH 2；
(31)Ac-Trp-Ala-Ala-Pro-Phe-Leu-Arg-Ala-OH；
(32)Ac-Trp-Ala-Ala-Pro-Phe-Leu-Arg-Ala-NH 2；
(33)H-Trp-Lys-Ala-Leu-Phe-Gln-Arg-Trp-OH；
(34)H-Trp-Lys-Gly-Pro-Tyr-Gln-Met-Trp-OH；
(35)Ac-Met-Leu-Leu-Leu-Ala-Asp-NH 2；
(36)Ac-Val-Asn-Gln-Leu-Val-Ala-NH 2；
(37)Ac-Met-Leu-Asp-Phe-Leu-Ala-NH 2；
(38)Ac-Leu-Trp-Phe-Val-Phe-Lys-NH 2；
(39)Ac-Glu-Thr-Gly-Trp-Ala-Ala-NH 2；
(40)Ac-Ala-Pro-Phe-Leu-Arg-Ala-NH 2；
(41)Ac-Pro-Gly-Asp-Ala-Tyr-Gly-NH 2；
(42)Ac-Tyr-Gly-Asp-Ala-Gly-Tyr-NH 2；
(43)Ac-Ser-Phe-Ser-Asn-Gln-Met-NH 2；
(44)Ac-Val-Asp-Cys-Ala-Asn-Lys-NH 2；
(45)Ac-Val-Phe-Ala-Phe-Leu-Tyr-NH 2；
(46)Ac-Ser-Ser-Ala-Trp-Ala-Lys-NH 2；
(47)Ac-Thr-Cys-Lys-Glu-Leu-Arg-NH 2；
(48)Ac-Leu-Asn-Leu-Val-Leu-Trp-NH 2。
进一步地，所述肽，或其立体异构体、或其立体异构体的混合物，或其美容上可接受的盐、或其药学上可接受的盐，选自Ac-Val-Lys-Val-Leu-Gln-Trp-NH 2、Ac-Ser-Met-Gly-Ala-Leu-Ala-NH 2或Ac-Trp-Ala-Ala-Pro-Phe-Leu-Arg-Ala-NH 2。
本申请的式(I)所示的肽可以作为立体异构体或立体异构体的混合物存在；例如，其所包含的这些氨基酸可以具有L-、D-的构型、或彼此独立地是外消旋的。因此，有可能获得同分异构混合物以及外消旋混合物或非对映混合物、或纯的非对映异构体或对映异构体，这取决于不对称碳的数量和存在什么同分异构体或同分异构混合物。本申请的式(I)所示的肽的优选的结构是纯的同分异构体，即，对映异构体或非对映异构体。
例如，当本申请所述-Phe-时，应理解-Phe-选自-L-Phe-、-D-Phe-、或两者的混合物，是外消旋的或非外消旋的。在本文件中描述的制备方法使本领域的普通技术人员能够通过选择具有正确构型的氨基酸来获得本申请的肽的每种立体异构体。
本申请还包括式(I)所示的肽的所有合适的同位素变体。本申请的这些肽的同位素变体此处理解为是指这样的化合物：其中在本申请的肽内至少一个原子被替换为相同原子序数的另一个原子，但 所述另一原子的原子质量不同于自然界中通常或主要存在的原子质量。可掺入本申请的肽中的同位素的实例是：氢、碳、氮、氧或硫的那些，例如 2H(氘)、 3H(氚)、 13C、 14C、 15N、 17O、 18O、 33S、 34S、 35S或 36S。本申请的肽的特定的同位素变体(特别是其中已经掺入一种或多种放射性同位素的那些)可能有利于，例如检查在体内的作用机理或活性化合物的分布；由于相对简单的可制备性和可检测性，尤其是用 3H或 14C同位素标记的化合物适用于该目的。另外，由于化合物的更强的代谢稳定性，同位素(例如氘)的掺入可以产生特定的治疗益处，例如体内半衰期的延长或所需活性剂量的降低；因此，在某些情况下，本申请的肽的这种改性还可构成本申请的优选实施方案。通过本领域技术人员已知的方法，例如通过在下文中进一步描述的方法和在实施例中所述的方法，通过使用各自的试剂和/或起始物质的相应的同位素改性物，可制备本申请的肽的同位素变体。
此外，本申请还包括本申请的肽的前药。术语“前药”在本文中意指这样的化合物：其本身可以是生物学上有活性的或无活性的，但是在它们在身体内的停留时间期间，其反应(例如代谢或水解)生成本申请的肽。
术语“美容上可接受的盐或药学上可接受的盐”指被认可的在动物，并且更确切地说在人类中使用的一种盐，包括式(I)所示的肽的金属盐，所述金属包括，但不局限于：锂、钠、钾、钙、镁、锰、铜、锌或铝等；包括式(I)所示的肽与有机碱形成的盐，所述有机碱包括：乙胺、二乙胺、乙二胺、乙醇胺、二乙醇胺、精氨酸、赖氨酸、组氨酸或哌嗪；包括式(I)所示的肽与无机酸或有机酸形成的盐，所述有机酸包括，但不局限于：乙酸、柠檬酸、乳酸、丙二酸、马来酸、酒石酸、延胡索酸、苯甲酸、天冬氨酸、谷氨酸、琥珀酸、油酸、三氟乙酸、草酸、扑酸(pamoate)或葡萄糖酸等；所述无机酸包括，但不局限于：盐酸、硫酸、硼酸或碳酸。
本申请式(I)所示的肽、或其立体异构体或其美容上可接受的盐、或其药学上可接受的盐的合成可以根据现有技术中已知的常规方法来进行，例如固相合成法、液相合成法或固相与液相结合的方法，还可以通过以产生所希望的序列为目标的生物技术方法、或通过具有动物、真菌、或植物来源的蛋白质的控制水解来制备。
例如，一种获得式(I)所示的肽的方法包括以下步骤：
-将具有受保护的N-末端和自由的C-末端的氨基酸与具有自由的N-末端和受保护的或与固体载体结合的C-末端的氨基酸偶联；
-消除保护N-末端的基团；
-重复该偶联顺序和消除保护N-末端的基团，直到获得所希望的肽序列；
-消除保护C-末端的基团或从该固体载体裂解。
优选地，C-末端与一种固体载体结合并且该方法是在固相上进行，包括将具有受保护的N-末端和自由的C-末端的氨基酸与具有自由的N-末端和与一种聚合物载体结合的C-末端的氨基酸偶联；消除保护N-末端的基团；并且重复此顺序所需要的次数以便因此获得具有所希望的长度的肽，接着从最初的聚合物载体裂解所合成的肽。
在整个合成中这些氨基酸的侧链的官能团用临时或永久的保护基团保持充分地保护，并且可以与从该聚合物载体裂解肽的过程同时地或正交地脱保护。
可选地，固相合成可以通过将二肽或三肽偶联到聚合物支持体上或偶联到先前与聚合物支持体结合的二肽或氨基酸上的汇集策略(convergent strategy)来进行。
使用本领域已知的标准条件和方法对N-末端和C-末端脱保护和/或以非确定的次序从聚合物支持体裂解肽，随后可以修饰所述末端的官能团。可以对与聚合物支持体结合的式(I)的肽进行N-末端和C-末端的任选的修饰，或在肽已从聚合物支持体裂解后进行N-末端和C-末端的任选的修饰。
本申请的另一方面，提供一种美容或药用组合物，包括有效量的上述式(I)所示的肽，或其立体异构体、或立体异构体的混合物，或其美容上可接受的盐、或其药学上可接受的盐，以及至少一种赋形剂和任选的美容上或药学上可接受的佐剂。
具体地，所述佐剂选自：胶原合成刺激剂、调节PGC-1α合成的剂、调节PPARγ的活性的剂、增加或减少脂肪细胞的甘油三酸酯含量的剂、刺激或延迟脂肪细胞分化的剂、脂解剂或刺激脂肪分解的剂、溶脂剂、生脂剂、乙酰胆碱受体聚集的抑制剂、抑制肌肉收缩的剂、抗胆碱能试剂、弹性蛋白酶抑制剂、基质金属蛋白酶抑制剂、黑色素合成刺激或抑制剂、增白剂或脱色剂、促色素沉着剂、自晒黑剂、抗老化剂、NO-合酶抑制剂、5α-还原酶抑制剂、赖氨酰羟化酶和/或脯氨酰羟化酶的抑制剂、抗氧化剂、自由基清除剂和/或抗大气污染的剂、活性羰基类物质清除剂、抗糖化剂、抗组胺剂、抗病毒剂、抗寄生虫剂、乳化剂、润肤剂、有机溶剂、液体推进剂、皮肤调理剂、保湿剂、保留水分的物质、α羟基酸、β羟基酸、增湿剂、表皮水解酶、维生素、氨基酸、蛋白质、色素或着色剂、染料、生物聚合物、胶凝聚合物、增稠剂、表面活性剂、软化剂、粘合剂、防腐剂、抗皱剂、能够减少或治疗下眼袋的剂、去角质剂、角质剥离剂、角质层分离剂、抗微生物剂、抗真菌剂、抑真菌剂、灭菌剂、抑菌剂、刺激真皮或表皮大分子的合成和/或能够抑制或预防它们的降解的剂、刺激弹性蛋白合成的剂、刺激核心蛋白聚糖合成的剂、刺激层粘连蛋白合成的剂、刺激防御素合成的剂、刺激伴侣蛋白合成的剂、刺激cAMP合成的剂、热休克蛋白、刺激HSP70合成的剂、刺激热休克蛋白合成的剂、刺激透明质酸合成的剂、刺激纤连蛋白合成的剂、刺激去乙酰化酶合成的剂、刺激脂质和角质层组分的合成的剂、神经酰胺、脂肪酸、抑制胶原降解的剂、抑制弹性蛋白降解的剂、抑制丝氨酸蛋白酶的剂、刺激成纤维细胞增殖的剂、刺激角质形成细胞增殖的剂、刺激脂肪细胞增殖的剂、刺激黑色素细胞增殖的剂、刺激角质形成细胞分化的剂、抑制乙酰胆碱酯酶的剂、皮肤松弛剂、刺激糖胺聚糖合成的剂、抗角化过度剂、粉刺溶解剂、抗银屑病剂、抗皮炎剂、抗湿疹剂、DNA修复剂、DNA防护剂、稳定剂、止痒剂、用于治疗和/或护理敏感性皮肤的剂、固化剂、紧致剂、重构剂、抗拉伸纹剂、粘合剂、调节皮脂产生的剂、止汗剂、刺激愈合的剂、协助愈合的剂、刺激再上皮化的剂、协助再上皮化的剂、细胞因子生长因子、镇静剂、抗炎剂、麻醉剂、作用于毛细血管循环和/或微循环的剂、刺激血管生成的剂、抑制血管渗透性的剂、静脉紧张剂、作用于细胞代谢的剂、用于改善真皮-表皮接合的剂、诱导毛发生长的剂、毛发生长抑制或延缓剂、香料、螯合剂、植物提取物、精油、海洋提取物、得自 生物发酵过程的剂、无机盐、细胞提取物、防晒剂、以及有效抗A和/或B紫外线的有机或无机光防护剂或其混合物。
进一步地，所述美容或药用组合物的制剂选自：霜剂、油、奶、香膏、泡沫、洗剂、凝胶、擦剂、浆液、皂、洗发精、润发乳、血清、软膏、摩丝、润发油、粉末、杆剂、笔剂、喷雾剂、气溶胶、胶囊剂、片剂、颗粒剂、口香糖、溶液、混悬液、乳剂、糖浆剂、酏剂、多糖薄膜、胶冻或明胶；
具体地，所述胶囊剂包括：软胶囊剂、硬胶囊剂，可选为明胶胶囊剂；所述片剂包括：糖衣片剂。
本申请的肽根据它们的序列的性质或N-末端和/或C-末端中的任何可能的修饰，在水中具有可变的溶解度。因此本申请的肽可以通过水溶液掺入组合物中，且不溶于水的那些可溶解于美容上或药学上可接受的常规溶剂中，所述溶剂例如并且不限于乙醇、丙醇、异丙醇、丙二醇、甘油、丁二醇或聚乙二醇或其任何组合。
待施用的美容上或药学上有效量的本申请的肽以及它们的剂量将依赖于许多因素，包括年龄、患者的状态、病症或疾病的严重性、施用的途径和频率以及待使用的肽的具体性质。
“美容上或药学上有效量”意指无毒性的但足以提供希望的效果的本申请的一种或多种肽的量。在本申请的美容组合物或药用组合物中以获得希望的效果的美容上或药学上有效的浓度使用本申请的肽；在一个优选形式中，相对于组合物的总重量，在0.00000001％(按重量计)和20％(按重量计)之间，优选在0.000001％(按重量计)和15％(按重量计)之间、更优选在0.0001％(按重量计)和10％(按重量计)之间，并且甚至更优选在0.0001％(按重量计)和5％(按重量计)之间。
本申请的另一方面，提供一种美容上或药学上可接受的递送系统或缓释系统，以便实现有效成分的更好渗透和/或改进它的药物代谢动力学和药效动力学特性，其包含有效量的上述式(I)所示的肽，或其立体异构体、或立体异构体的混合物，或其美容上可接受的盐、或其药学上可接受的盐，或上述的美容或药用组合物。
术语“递送系统”是指与本申请的肽一起施用的稀释剂、佐剂、赋形剂或载体，它们选自：水、油或表面活性剂、包括石油来源、动物来源、植物来源、或合成来源的那些，例如并且不限于花生油、大豆油、矿物油、芝麻油、蓖麻油、聚山梨醇酯、脱水山梨糖醇酯、醚硫酸酯、硫酸酯、甜菜碱、葡萄糖苷、麦芽糖苷、脂肪醇、壬苯醇醚、泊洛沙姆、聚氧乙烯、聚乙二醇、右旋糖、甘油、毛地黄皂苷和类似物。本领域的普通技术人员已知在可以给予本申请的肽的不同递送系统中可以使用的稀释剂。
术语“缓释”以常规含义使用，指提供化合物在一段时间内逐渐释放的化合物的递送系统，且优选地但不是必须地，在整个时间段内具有相对恒定的化合物释放水平。
递送系统或缓释系统的实例是脂质体、油质体、非离子型表面活性剂脂质体囊泡、醇质体、毫米胶囊、微米胶囊、纳米胶囊、纳米结构的脂质载体、海绵状物、环糊精、类脂囊泡、胶束、毫米球、微米球、纳米球、脂质球、微米乳液、纳米乳液、毫米粒子、微米粒子或纳米粒子。优选的递送系统或缓释系统是脂质体和微米乳液，更优选具有反胶束的内部结构的油包水型微米乳液。
缓释系统可以通过现有技术中已知的方法来制备，并且可以例如通过以下方式来给予：通过局部 或经皮给药，包括粘附贴剂、非粘附贴剂、封闭贴剂、以及微电子贴剂；或通过全身给药例如并且不局限于，口服或胃肠外途径，包括鼻、直肠、皮下植入或注射、或直接植入或注射至特定身体部位中，并且优选地应该释放相对恒定量的本申请的这些肽。在该缓释系统中包含的肽的量将取决于例如该组合物将被给予的部位、本申请的肽的释放动力学和持续时间、以及有待治疗和/或护理的病状、病症和/或疾病的性质。
本申请的另一方面，提供一种上述式(I)所示的肽，或其立体异构体、或立体异构体的混合物，或其美容上可接受的盐、或其药学上可接受的盐，或上述美容或药用组合物，或上述的美容上或药学上可接受的递送系统或缓释系统在制备用于治疗或护理由肌肉收缩导致的病况、失调或疾病的美容组合物或药用组合物中的用途，其中，所述失调或疾病是张力障碍。
本申请的实施方案，所述失调或疾病是张力障碍，具体地，所述张力障碍为局灶性肌张力障碍；张力障碍包括，但不限于脸痉挛、扭转性张力障碍、颈张力障碍或斜颈症、喉张力障碍或痉挛性发音困难、口下颌张力障碍、肢体张力障碍诸如书写痉挛或音乐家痉挛或脚部张力障碍、夜间磨牙、半侧颜面痉挛、抽搐症和/或斜视；节段性肌张力障碍、梅杰氏综合征、多灶性张力障碍、偏侧肌张力障碍、多巴胺反应性肌张力障碍和濑川张力障碍。
本申请的另一方面，提供一种上述式(I)所示的肽，或其立体异构体、或立体异构体的混合物，或其美容上可接受的盐、或其药学上可接受的盐，或上述美容或药用组合物，或上述的美容上或药学上可接受的递送系统或缓释系统在制备用于治疗、预防或修复皮肤老化或光老化的美容组合物或药用组合物中的用途，其中，所述皮肤老化或光老化的治疗、预防或修复是减少、预防或治疗面部皱纹。
为了便于理解本申请，对在本申请所使用的一些术语和表述的含义说明如下：
在本申请中，术语“皮肤”应理解为是构成它的多个层，从最上层或角质层至最下层或皮下组织，两个端点都包括在内。这些层由不同类型的细胞组成，如角质形成细胞、成纤维细胞、黑色素细胞、和/或脂肪细胞等。在本申请中，术语“皮肤”包括头皮。
术语“治疗”，指的是给予根据本申请的肽以减轻或消除一种疾病或病症、或减少或消除与这种疾病或病症相关的一种或多种症状。术语“治疗”还涵盖了减轻或消除该疾病或病症的生理后果的能力。
术语“护理”包括疾病和/或病症的预防。
术语“预防”，指的是本申请的肽在一种疾病或病症出现前防止、延迟、或阻碍其出现或发展的能力。
术语“老化”指的是皮肤随着年龄的增长经历的变化(自然老化)，或通过暴露于阳光(光老化)或暴露于环境污染物，如化学污垢或污染物、烟草烟雾等而经历的变化，并且包括所有外在可见的和/或通过触摸可感知的变化，例如并且不局限于：皮肤上的不连续性的发展(如皱纹、细纹、表情纹、拉伸纹、条纹、沟纹、不平整或粗糙、毛孔尺寸增大、水分损失、弹性损失、紧致性损失、平滑性损失、变形恢复能力损失、回弹性损失)、皮肤下垂(如脸颊下垂、眼睛下方出现眼袋、或出现双下巴 等)、皮肤颜色的变化(如瘢痕、变红、眼袋、或出现色素过度沉着区域如老年斑或雀斑等)、异常分化、过度角质化、弹性组织变性、角化症、脱发、橘皮样皮肤、胶原结构损失，以及角质层、真皮、表皮、血管系统(例如出现蜘蛛静脉或毛细血管扩张症)或靠近皮肤的那些组织的其他组织学变化。
术语“光老化”指的是由于皮肤长期暴露于紫外线辐射而导致的皮肤过早老化，它呈现出与自然老化相同的生理特征，例如并且不局限于：松弛、下垂、颜色改变或色素沉着不规则、异常和/或过度角质化。
在本说明书中，用于氨基酸的缩写遵循IUPAC-IUB生化命名委员会(IUPAC-IUB Commission of Biochemical Nomenclature)在欧洲生物化学杂志(Eur.J.Biochem.1984,138:9-37)中所指定的规则。
因此，例如，Lys表示NH 2-CH(CH 2CH 2CH 2CH 2NH 2)-COOH，Lys-表示NH 2-CH(CH 2CH 2CH 2CH 2NH 2)-CO-，-Lys表示-NH-CH(CH 2CH 2CH 2CH 2NH 2)-COOH，并且-Lys-表示-NH-CH(CH 2CH 2CH 2CH 2NH 2)-CO-。因此，表示肽键的连字符消除了当位于该符号的右侧时的氨基酸(在此用常规非离子化形式来表示)1-羧基中的OH，并且消除了当位于该符号的左侧时的氨基酸2-氨基中的H。
缩写“Ac-”在本申请中用来表示乙酰基(CH 3-CO-)；Ser：丝氨酸；Gly：甘氨酸；Ala：丙氨酸；Lys：赖氨酸；Trp：色氨酸；Val：缬氨酸；Leu：亮氨酸；Gln：谷氨酰胺；Met：甲硫氨酸；His：组氨酸；Arg：精氨酸；Pro：脯氨酸；Phe：苯丙氨酸；Tyr：酪氨酸。
本申请具有以下优点和效果：
本申请多肽通过人工设计得到，合成方便，通过与突触小泡缔合性膜蛋白(VAMP2)的相互作用，调节SNARE复合物的形成，从而干扰突触神经递质释放，抑制肌肉收缩，改善皮肤表面的皱纹，减少皱纹深度，达到祛皱效果，可以在化妆品或医药领域用于改善皮肤老化或光老化，治疗或护理由肌肉收缩导致的病况、失调或疾病。
附图说明
为了更清楚地说明本申请的技术方案，下面将对本申请的描述中所需要使用的附图作简单地介绍，显而易见地，下面描述中的附图仅仅是本申请的一些实施例，对于本领域普通技术人员来讲，在不付出创造性劳动的前提下，还可以根据这些附图获得其他的附图。
图1是肽(4)Ac-Val-Lys-Val-Leu-Gln-Trp-NH 2(分子式C 40H 64N 10O 8)质谱图，[M+H] +准分子离子峰的质荷比(m/z)为813.5879，质谱测得的分子量为812.59；
图2是肽(8)Ac-Ser-Met-Gly-Ala-Leu-Ala-NH 2(分子式C 24H 43N 7O 8S)质谱图，[M+Na] +加和离子峰的质荷比(m/z)为612.3438，质谱测得的分子量为589.34；
图3是肽(32)Ac-Trp-Ala-Ala-Pro-Phe-Leu-Arg-Ala-NH 2(分子式C 48H 69N 13O 9)质谱图，[M+H] +准分子离子峰的质荷比(m/z)为972.6496，质谱测得的分子量为971.65；
图4是测试样品肌肉收缩抑制实验的结果图。
具体实施方式
为使本申请的所述目的、特征和优点能够更加明显易懂，下面结合附图和实施例对本申请作进一 步详细的说明。显然，所描述的实施例是本申请一部分实施例，而不是全部的实施例。基于本申请中的实施例，本领域普通技术人员在没有作出创造性劳动前提下所获得的所有其他实施例，都属于本申请保护的范围。
本申请的核心构思之处在于，基于Synaptophysin得到了一种仿生肽，这些肽可以干扰Synaptophysin与VAMP2相互作用，并因此抑制SNARE复合物的形成和神经递质释放，从而抑制肌肉收缩。所述多肽具有以下通式：R 1-X 1-X 2-X 3-X 4-X 5-X 6-X 7-X 8-R 2；本申请的肽、其立体异构体、立体异构体的混合物、或其美容上可接受的盐、或其药学上可接受的盐、或它们的美容组合物或药用组合物、以及它们在制备用于改善皮肤老化或光老化，治疗或护理由肌肉收缩导致的病况、失调或疾病的美容组合物或药物组合物中的用途。
需要说明的是，在本申请中，用于氨基酸的缩写遵循IUPAC-IUB的生物化学命名委员会在Eur.J.Biochem.(1984)138:9-37和J.Chem.(1989)264:633-673中指定的规则。
Amide Resin：一种多肽合成用的起始树脂(交联度1％，替代度1.72mmol/g，粒度100-200目)；Fmoc-Linker：4-[(2,4-二甲氧基苯基)(Fmoc-氨基)甲基]苯氧乙酸；Ac 2O：醋酸酐；DMF：N,N-二甲基甲酰胺；DIPEA：二异丙基乙胺；DIC：二异丙基碳二亚胺；piperidine：哌啶；HOBt：1-羟基苯并三氮唑；TFA：三氟乙酸；TIS：三异丙基硅烷；EDT：1,2-乙二硫醇；Ser：丝氨酸；Gly：甘氨酸；Ala：丙氨酸；Lys：赖氨酸；Trp:色氨酸；Val：缬氨酸；Leu：亮氨酸；Gln：谷氨酰胺；Met：甲硫氨酸；Arg：精氨酸；Pro：脯氨酸；Phe：苯丙氨酸；Fmoc：9-芴基甲氧羰基；Boc：叔丁氧基羰基；tBu：叔丁基；Trt：三苯甲基；Pbf：2,2,4,6,7-五甲基二氢苯并呋喃-5-磺酰基。
实施例1计算机模拟
在该实验中，利用计算机模拟方法筛选出候选肽，其对Synaptophysin与VAMP2相互作用区域具有亲和力或特异性，因此，能够干扰Synaptophysin与VAMP2之间的相互作用或结合，进而影响SNARE复合物的形成。
首先以Synaptophysin的全序列为目的蛋白，从中截取不同长度的氨基酸残基片段，构建肽链长度分别为5、6、7和8个AAs的重叠肽库。由于SNARE复合物中VAMP2为螺旋束结构，α-helic对多肽与VAMP2的亲和力具有重要影响，因此利用计算机分析多肽的α-helic性质。
根据计算机模拟多肽的α-helic性质结果，初步筛选出27条多肽，见下表1。
表1 利用计算机模拟初步筛选出的多肽
Figure PCTCN2022083491-appb-000001
Figure PCTCN2022083491-appb-000002
为了进一步评估多肽与VAMP2的结合能力，选择参与形成SNARE复合物之前的VAMP2进行研究。利用分子对接和分子动力学计算上述多肽与VAMP2的亲和力，进而筛选出最佳候选肽，并考虑对它们做进一步活性验证。最佳候选肽见下表2。
表2 利用计算机模拟筛选出的最佳候选多肽
Figure PCTCN2022083491-appb-000003
结果显示，本申请的肽可以与VAMP2较好地结合，从而干扰SNARE复合物的形成，可用于抑制肌肉收缩。
实施例2
Ac-Val-Lys-Val-Leu-Gln-Trp-NH 2的制备
2.1 Fmoc-Linker-Amide Resin的制备
称取5g Amide Resin树脂于固相合成反应柱中，用DMF溶胀，洗涤树脂，抽走溶剂。
称取7g的Fmoc-Linker、2.10g的HOBt于干燥三角瓶中。用DMF溶剂溶解后置于冰水浴中冷却10min，加DIC活化10min，避免水汽。
将活化后的Fmoc-Linker加入溶胀后的树脂中反应2.5h，抽走反应液，洗涤树脂，抽走溶剂。
继续加入Ac 2O与DIPEA封端处理1.5h。洗涤树脂，抽走溶剂。
2.2脱Fmoc
Fmoc-Linker-Amide Resin用20％piperidine/DMF脱Fmoc二次，每次10min，取样K检，显色深蓝。用DMF洗涤树脂7次，抽走溶剂。
2.3投料反应
称取5.3g的Fmoc-Trp(Boc)-OH，1.8g的HOBt加入干燥三角瓶中，加入DMF使其溶解，密封置于-18℃冰箱30min。加1.65g DIC活化3min，避免水汽。将活化后的氨基酸加入脱保护后树脂中反应2h，抽走反应液。K检树脂无色透明说明反应完全。
对N-末端Fmoc基团进行脱保护，并且在存在1.8g HOBt和1.65g DIC的情况下，使用DMF作为溶剂，将活化后的7.3g Fmoc-Gln(Trt)-OH偶联至肽基树脂上，持续反应2h。然后洗涤这些树脂并且重复Fmoc基团的脱保护处理以便偶联下一个氨基酸。在每次偶联中，在存在1.8g HOBt和1.65g DIC的情况下，使用DMF作为溶剂，顺序地偶联4.2g的Fmoc-Leu-OH；4.0g的Fmoc-Val-OH；5.6g的Fmoc-Lys(Boc)-OH以及随后4.0g的Fmoc-Val-OH；反应完全之后，洗涤树脂，抽走溶剂。
对肽基树脂的N-末端Fmoc基团脱保护，用20％piperidine/DMF脱Fmoc二次，每次10min，取样K检，显色深蓝。用DMF洗涤树脂6次，抽走溶剂。
在存在DIPEA的情况下，使用DMF作为溶剂，将1.9g的Ac 2O偶联至肽基树脂上，持续反应1.5h，洗涤树脂，抽走溶剂，收缩干燥后得到12g的Ac-Val-Lys(Boc)-Val-Leu-Gln(Trt)-Trp(Boc)-Linker-Amide Resin。
2.4裂解
量取54mL的TFA、1.5mL的TIS、1.5mL的EDT、1.5mL的苯甲硫醚、1.5mL的水，混合搅拌均匀后得到裂解液，封口放置-18℃冰箱备用；异丙醚放置于-18℃冰箱冷冻备用。
称取12g Ac-Val-Lys(Boc)-Val-Leu-Gln(Trt)-Trp(Boc)-Linker-Amide Resin，加入圆底烧瓶中，加入上述冷冻好的裂解液，搅拌反应2h。抽滤，收集滤液浓缩到10mL后加入异丙醚搅拌离心洗涤6次，直至pH值为3-4，真空干燥，得到4.5g的Ac-Val-Lys-Val-Leu-Gln-Trp-NH 2粗肽。
2.5纯化
称取4.5g粗肽溶于纯水中，浓缩后加入甲醇使其全部溶解，用孔径为0.22μm微孔滤膜过滤得到澄清透明溶液，通过反相HPLC纯化处理，纯化梯度如下表：
时间(min) 流速(mL/min) A％(0.1％醋酸+乙腈) B％(0.1％醋酸+纯水)
0 40 10 90
15 40 30 70
50 40 45 55
将过滤后的样品进样纯化，收集馏分，浓缩冻干，得到纯度98％的肽(4)Ac-Val-Lys-Val-Leu-Gln-Trp-NH 2。
实施例3
Ac-Ser-Met-Gly-Ala-Leu-Ala-NH 2的制备
3.1 Fmoc-Linker-Amide Resin的制备
称取5g Amide Resin树脂于固相合成反应柱中，用DMF溶胀，洗涤树脂，抽走溶剂。
称取6.9g的Fmoc-Linker、2.0g的HOBt于干燥三角瓶中。用DMF溶剂溶解后置于冰水浴中冷却10min，加DIC活化10min，避免水汽。
将活化后的Fmoc-Linker加入溶胀后的树脂中反应2.5h，抽走反应液，洗涤树脂，抽走溶剂。
继续加入Ac 2O与DIPEA封端处理1.5h。洗涤树脂，抽走溶剂。
3.2脱Fmoc
Fmoc-Linker-Amide Resin用20％piperidine/DMF脱Fmoc二次，每次10min，取样K检，显色深蓝。用DMF洗涤树脂8次，抽走溶剂。
3.3投料反应
称取3.1g的Fmoc-Ala-OH，1.8g的HOBt加入干燥三角瓶中，加入DMF使其溶解，密封置于-18℃冰箱30min。加1.65g DIC活化3min，避免水汽。将活化后的氨基酸加入脱保护后树脂中反应2h，抽走反应液。K检树脂无色透明说明反应完全。
对N-末端Fmoc基团进行脱保护，并且在存在1.8g HOBt和1.65g DIC的情况下，使用DMF作为溶剂，将活化后的4.2g Fmoc-Leu-OH偶联至肽基树脂上，持续反应2h。然后洗涤这些树脂并且重复Fmoc基团的脱保护处理以便偶联下一个氨基酸。在每次偶联中，在存在1.8g HOBt和1.65g DIC的情况下，使用DMF作为溶剂，顺序地偶联3.1g的Fmoc-Ala-OH；3.6g的Fmoc-Gly-OH；4.5g的 Fmoc-Met-OH以及随后4.6g的Fmoc-Ser(tBu)-OH；反应完全之后，洗涤树脂，抽走溶剂。
对肽基树脂的N-末端Fmoc基团脱保护，用20％piperidine/DMF脱Fmoc二次，每次10min，取样K检，显色深蓝。用DMF洗涤树脂6次，抽走溶剂。
在存在DIPEA的情况下，使用DMF作为溶剂，将1.9g的Ac 2O偶联至肽基树脂上，持续反应1.5h，洗涤树脂，抽走溶剂，收缩干燥后得到13g的Ac-Ser(tBu)-Met-Gly-Ala-Leu-Ala-Linker-Amide Resin。
3.4裂解
量取44mL的TFA、1.2mL的TIS、1.2mL的EDT、1.2mL的水混合搅拌均匀后得到裂解液，封口放置-18℃冰箱备用；异丙醚放置于-18℃冰箱冷冻备用。
称取13g Ac-Ser(tBu)-Met-Gly-Ala-Leu-Ala-Linker-Amide Resin，加入圆底烧瓶中，加入上述冷冻好的裂解液，搅拌反应2h。抽滤，收集滤液浓缩到10mL后加入异丙醚搅拌离心洗涤6次，直至pH值为3-4，真空干燥，得到6.9g的Ac-Ser-Met-Gly-Ala-Leu-Ala-NH 2粗肽。
3.5纯化
称取6.9g粗肽溶于370mL纯水中，用孔径为0.22μm微孔滤膜过滤得到澄清透明溶液，通过反相HPLC纯化处理，纯化梯度如下表：
时间(min) 流速(mL/min) A％(0.1％醋酸+乙腈) B％(0.1％醋酸+纯水)
0 40 10 90
15 40 18 82
60 40 38 62
将过滤后的样品进样纯化，收集馏分，浓缩冻干，得到纯度99.4％的肽(8)Ac-Ser-Met-Gly-Ala-Leu-Ala-NH 2。
实施例4
Ac-Trp-Ala-Ala-Pro-Phe-Leu-Arg-Ala-NH 2的制备
4.1 Fmoc-Linker-Amide Resin的制备
称取5g Amide Resin树脂于固相合成反应柱中，用DMF溶胀，洗涤树脂，抽走溶剂。
称取6.9g的Fmoc-Linker、2.0g的HOBt于干燥三角瓶中。用DMF溶剂溶解后置于冰水浴中冷却10min，加DIC活化10min，避免水汽。
将活化后的Fmoc-Linker加入溶胀后的树脂中反应2.5h，抽走反应液，洗涤树脂，抽走溶剂。
继续加入Ac 2O与DIPEA封端处理1.5h。洗涤树脂，抽走溶剂。
4.2脱Fmoc
Fmoc-Linker-Amide Resin用20％piperidine/DMF脱Fmoc二次，每次10min，取样K检，显色深蓝。用DMF洗涤树脂7次，抽走溶剂。
4.3投料反应
称取3.1g的Fmoc-Ala-OH，1.8g的HOBt加入干燥三角瓶中。加入DMF使其溶解，密封置于-18℃冰箱30min。加1.65g的DIC活化3min，避免水汽。将活化后的氨基酸加入脱保护后树脂中反应2h， 抽走反应液。K检树脂无色透明说明反应完全。
对N-末端Fmoc基团进行脱保护，并且在存在1.8g HOBt和1.65g DIC的情况下，使用DMF作为溶剂，将活化后的7.6g Fmoc-Arg(Pbf)-OH偶联至肽基树脂上，持续反应2h。然后洗涤这些树脂并且重复Fmoc基团的脱保护处理以便偶联下一个氨基酸。在每次偶联中，在存在1.8g HOBt和1.65g DIC的情况下，使用DMF作为溶剂，顺序地偶联4.2g的Fmoc-Leu-OH；5.3g的Fmoc-Phe-OH；4.62g的Fmoc-Pro-OH；4.26g的Fmoc-Ala-OH；4.26g的Fmoc-Ala-OH以及随后9.26g的Fmoc-Trp(Boc)-OH；反应完全之后，洗涤树脂，抽走溶剂。
对肽基树脂的N-末端Fmoc基团脱保护，用20％piperidine/DMF脱Fmoc二次，每次10min，取样K检，显色深蓝。用DMF洗涤树脂6次，抽走溶剂。
在存在DIPEA的情况下，使用DMF作为溶剂，将1.9g的Ac 2O偶联至肽基树脂上，持续反应1.5h，洗涤树脂，抽走溶剂，收缩干燥后得到20.3g的Ac-Trp(Boc)-Ala-Ala-Pro-Phe-Leu-Arg(Pbf)-Ala-Linker-Amide Resin。
4.4裂解
量取114mL的TFA、3mL的TIS、3mL的水混合搅拌均匀后得到裂解液，封口放置-18℃冰箱备用；异丙醚放置于-18℃冰箱冷冻备用。
称取20.3g Ac-Trp(Boc)-Ala-Ala-Pro-Phe-Leu-Arg(Pbf)-Ala-Linker-Amide Resin，加入圆底烧瓶中，加入上述冷冻好的裂解液，搅拌反应2h。抽滤，收集滤液浓缩到10mL后加入异丙醚搅拌离心洗涤6次，直至pH值为3-4，真空干燥。得到9.3g的Ac-Trp-Ala-Ala-Pro-Phe-Leu-Arg-Ala-NH 2粗肽。
4.5纯化
称取9.3g粗肽溶于100mL甲醇中，加入500mL纯水，浓缩后，补加100mL甲醇，用孔径为0.22μm微孔滤膜过滤得到澄清透明溶液，通过反相HPLC纯化处理，纯化梯度如下表：
时间(min) 流速(mL/min) A％(0.1％醋酸+乙腈) B％(0.1％醋酸+纯水)
0 40 20 80
15 40 30 70
50 40 45 55
将过滤后的样品进样纯化，收集馏分，浓缩冻干，得到纯度98％的肽(32)Ac-Trp-Ala-Ala-Pro-Phe-Leu-Arg-Ala-NH 2。
实施例5
本申请式(I)中的其他化合物可以通过类似的方法制备。
所获得的这些肽通过ESI-MS测定其分子量，部分化合物的测试结果见下表3及图1-3：
表3 质谱法测定分子量结果
编号 序列 分子量质谱分析结果
(4) Ac-Val-Lys-Val-Leu-Gln-Trp-NH 2 812.59
(8) Ac-Ser-Met-Gly-Ala-Leu-Ala-NH 2 589.34
(32) Ac-Trp-Ala-Ala-Pro-Phe-Leu-Arg-Ala-NH 2 971.65
实施例6
肌肉收缩抑制实验
6.1实验动物
10周龄雌性SD大鼠，重量220g-260g。
6.2实验仪器
HW-400S恒温平滑肌槽、BL-420E生物机能实验系统、张力换能器，其他仪器为常规实验仪器。
6.3实验试剂
氯化乙酰胆碱(上海源叶)、水合氯醛(上海麦克林)
6.4待测样品
乙酰基八肽-3(Ac-Glu-Glu-Met-Gln-Arg-Arg-Ala-Asp-NH 2)，测试浓度分别为25ppm、50ppm、100ppm、200ppm；
乙酰基六肽-30(Ac-Glu-Asp-Tyr-Tyr-Arg-Leu-NH 2)，测试浓度分别为25ppm、50ppm、100ppm、200ppm；
肽(4)，测试浓度分别为25ppm、50ppm、100ppm、200ppm；
肽(8)，测试浓度分别为25ppm、50ppm、100ppm、200ppm；
肽(32)，测试浓度分别为100ppm、200ppm、400ppm、800ppm。
6.5实验方法
将实验大鼠实验前禁食不禁水12小时。按0.1mL/25g剂量腹腔注射10％水合氯醛，待大鼠麻醉后，剪开腹腔，露出膨大盲肠，距回盲部肠处取出一段回肠，在预先通氧10min的台式液中洗干净，剪取数段4cm长回肠孵育于4℃台式液中备用；在回肠两端分别用棉线部分结扎并打结使之成一环；
打开HW-400S恒温平滑肌槽、BL-420E生物机能实验系统，将张力换能器连接至通道1，调节孵育温度为37℃，仪器预热0.5小时，通氧10min以上。肠段一端用棉线固定在进气支架口，另一端与张力换能器相连，适当调节换能器的位置与高度，使离体肠段置于平滑肌槽中央，开始记录肠活动情况；
待肌肉收缩稳定后，加入终浓度0.1-1μg/mL的乙酰胆碱，直至肌肉活动出现高位而稳定的波形，作用5分钟作为初始对照；平衡5分钟后，分别加入待测样品。每更换一种受试药物，换新的回肠段，仪器记录相关数据，根据下列公式计算回肠平滑肌收缩抑制率：
回肠平滑肌收缩抑制率＝(乙酰胆碱给药后收缩力-待测样品给药后收缩力)/乙酰胆碱给药后收缩力×100％
实验数据以均数标准差(Mean±SD)表示。
6.6实验结果
测试样品的回肠平滑肌收缩抑制率结果见下表4及图4。
表4 测试样品的回肠平滑肌收缩抑制率(％)
Figure PCTCN2022083491-appb-000004
结果表明，给予乙酰胆碱后，能产生一段高位信号峰，给予乙酰基六肽-30在200ppm后该峰明显减弱，产生抑制作用，在200ppm的抑制率为48.09％；给予乙酰基八肽-3在50ppm时开始对乙酰胆碱产生的高位信号峰产生抑制作用，其在50ppm的抑制率为32.34％，在100ppm的抑制率为47.49％，在200ppm的抑制率为58.67％；然而给予肽(4)和肽(8)在25ppm时即对乙酰胆碱产生的高位信号峰产生明显的抑制作用，且随浓度升高，抑制作用随之增强；肽(4)在25ppm产生抑制效果，抑制率为49.78％，在50ppm的抑制率为62.40％，在100ppm的抑制率为66.72％，在200ppm浓度下抑制率高达76.88％；肽(8)在25ppm抑制率为31.99％，在50ppm的抑制率为34.35％，在100ppm的抑制率为56.42％，200ppm浓度下抑制率高达67.96％；给予肽(32)在800ppm对肌肉收缩产生抑制作用，抑制率为44.41％。
由此可知，本申请的肽具有明显的抑制肌肉收缩的作用，且比现有技术多肽的作用更强；其抑制有效率为肽(4)＞肽(8)＞乙酰基八肽-3＞乙酰基六肽-30。
抑制面部表情肌的收缩可以有效地改善眼角纹、法令纹等面部皱纹。乙酰胆碱作为神经递质，能直接激动平滑肌上的乙酰胆碱受体，触发信号级联反应而引起平滑肌产生收缩效应。通过抑制细胞乙酰胆碱释放或阻止乙酰胆碱与其受体结合，可以抑制肌肉收缩，治疗或护理由肌肉收缩导致的病况、失调或疾病，治疗、预防或修复皮肤老化或光老化，减少、预防或治疗面部皱纹。本申请的肽通过干扰Synaptophysin与VAMP2相互作用，影响VAMP2与SNAP25和Syntaxin形成SNARE复合物，进而抑制乙酰胆碱释放，因此抑制肌肉收缩。本实验通过检测回肠平滑肌收缩信号，比较各样品的回肠平滑肌收缩抑制率，进一步阐明了本申请的肽所具有的抑制肌肉收缩的技术效果。
实施例7
含肽(4)的乳液的制备
Figure PCTCN2022083491-appb-000005
Figure PCTCN2022083491-appb-000006
配制方法：将肽(4)配制为0.02mg/mL的水溶液；将鲸蜡硬脂醇(和)鲸蜡硬脂基葡糖苷、霍霍巴油、矿物油、棕榈酸异丙酯加热至85℃，搅拌均匀；得A相；将甘油、尿囊素、聚丙烯酰胺(和)C13-14异链烷烃(和)月桂醇聚醚-7用水溶解，加热至85℃，得B相；将A相快速加入B相，恒温均质3-5min，冷却；冷却至60℃以下加入防腐剂，搅拌均匀，冷却至45℃以下加入多肽溶液、香精，即得。
实施例8
含肽(8)的精华液的制备
Figure PCTCN2022083491-appb-000007
配制方法：将处方量的透明质酸钠加入水中，搅拌使混合均匀，然后加热到80～85℃，保温搅拌使其分散均匀；温度降到40℃以下，加入甘油、芦荟胶、肽(8)、维生素C、辛甘醇和1,2-己二醇，搅拌均匀；用三乙醇胺溶液调节溶液pH值至5.5左右，即得。
尽管已描述了本申请实施例的优选实施例，但本领域内的技术人员一旦得知了基本创造性概念，则可对这些实施例做出另外的变更和修改。所以，所附权利要求意欲解释为包括优选实施例以及落入 本申请实施例范围的所有变更和修改。
最后，还需要说明的是，在本文中，诸如第一和第二等之类的关系术语仅仅用来将一个实体或者操作与另一个实体或操作区分开来，而不一定要求或者暗示这些实体或操作之间存在任何这种实际的关系或者顺序。而且，术语“包括”、“包含”或者其任何其他变体意在涵盖非排他性的包含，从而使得包括一系列要素的过程、方法、物品或者终端设备不仅包括那些要素，而且还包括没有明确列出的其他要素，或者是还包括为这种过程、方法、物品或者终端设备所固有的要素。在没有更多限制的情况下，由语句“包括一个……”限定的要素，并不排除在包括所述要素的过程、方法、物品或者终端设备中还存在另外的相同要素。
以上对本申请所提供的一种多肽及其美容组合物或药用组合物和用途，进行了详细介绍，本文中应用了具体个例对本申请的原理及实施方式进行了阐述，以上实施例的说明只是用于帮助理解本申请的方法及其核心思想；同时，对于本领域的一般技术人员，依据本申请的思想，在具体实施方式及应用范围上均会有改变之处，综上所述，本说明书内容不应理解为对本申请的限制。

Claims (12)

  1. 一种式(I)所示的肽，或其立体异构体、或其立体异构体的混合物，或其美容上可接受的盐、或其药学上可接受的盐，其特征在于，
    R 1-X 1-X 2-X 3-X 4-X 5-X 6-X 7-X 8-R 2  (I)
    式(I)中，
    X 1选自：-Val-、-Ser-、-Trp-、-Met-、-Leu-、-Glu-、-Ala-、-Pro-、-Tyr-或-Thr-；
    X 2选自：-Lys-、-Arg-、-His-、-Met-、-Ala-、-Leu-、-Asn-、-Trp-、-Thr-、-Pro-、-Gly-、-Phe-、-Asp-、-Ser-或-Cys-；
    X 3选自：-Val-、-Gly-、-Leu-、-Pro-、-Met-、-Ala-、-Gln-、-Asp-、-Phe-、-Ser-、-Cys-或-Lys-；
    X 4选自：-Gly-、-Leu-、-Pro-、-Ala-、-Phe-、-Val-、-Trp-、-Asn-或-Glu-；
    X 5选自：-Phe-、-Trp-、-Tyr-或键；
    X 6选自：-Gln-、-Leu-、-Ala-、-Val-、-Phe-、-Arg-、-Tyr-、-Gly-、或-Asn-；
    X 7选自：-Lys-、-Arg-、-His-或键；
    X 8选自：-Trp-、-Ala-、-Asp-、-Lys-、-Gly-、-Tyr-、-Met-或-Arg-；
    R 1选自：H或R 3-CO-，其中R 3选自：取代的或未取代的烷基、取代的或未取代的烯基；
    R 2选自：-NR 4R 5或-OR 4，其中各个R 4和R 5彼此独立地选自：H、取代的或未取代的烷基、取代的或未取代的烯基；
    所述烷基是指具有1-24个碳原子(可选具有1-16个碳原子；可选具有1-14个碳原子；可选具有1-12个碳原子；可选具有1、2、3、4、5、或6个的碳原子)的饱和脂肪族直链或支链的烷基；可选选自：甲基、乙基、异丙基、异丁基、叔丁基、戊基、己基、庚基、辛基、癸基、十二烷基、十四烷基、十六烷基、十八烷基、2-乙基己基、2-甲基丁基、或5-甲基己基；
    所述烯基是指具有2-24个碳原子(可选具有2-16个碳原子；可选具有2-14个碳原子；可选具有2-12个碳原子；可选具有2、3、4、5、或6个碳原子)的直链或支链烯基；所述烯基具有一个或多个碳-碳双键，可选具有1、2或3个共轭或非共轭的碳-碳双键；所述烯基是通过一个单键而结合至分子的其余部分；可选选自：乙烯基、油烯基、或亚油烯基；
    可选地，所述“取代的烷基”、“取代的烯基”中的取代基选自C 1-C 4烷基；羟基；C 1-C 4烷氧基；氨基；C 1-C 4氨基烷基；C 1-C 4羰氧基；C 1-C 4氧基羰基；卤素(如氟、氯、溴、以及碘)；氰基；硝基；叠氮化物；C 1-C 4烷基磺酰基；硫醇；C 1-C 4烷硫基；C 6-C 30芳氧基如苯氧基；-NR b(C＝NR b)NR bR c，其中R b和R c是独立地选自：H、C 1-C 4烷基、C 2-C 4烯基、C 2-C 4炔基、C 3-C 10环烷基、C 6-C 18芳基、C 7-C 17芳烷基、具有三至十元的杂环基、或氨基的保护基。
  2. 根据权利要求1所述的式(I)所示的肽，或其立体异构体、或其立体异构体的混合物，或其美容上可接受的盐、或其药学上可接受的盐，其特征在于，R 1选自：H、乙酰基、叔-丁酰基、己酰基、2-甲基己酰基、辛酰基、癸酰基、月桂酰基、肉豆蔻酰基、棕榈酰基、硬脂酰基、油酰基或亚油酰基；
    其中，R 1选自H、乙酰基、月桂酰基、肉豆蔻酰基或棕榈酰基；
    可选地，R 1是H或乙酰基。
  3. 根据权利要求1所述的式(I)所示的肽，或其立体异构体、或其立体异构体的混合物，或其美容上可接受的盐、或其药学上可接受的盐，其特征在于，R 4、R 5彼此独立地选自：H、甲基、乙基、己基、十二烷基或十六烷基；
    其中，R 4是H并且R 5选自：H、甲基、乙基、己基、十二烷基或十六烷基；
    可选地，R 2是-OH或-NH 2。
  4. 根据权利要求1所述的式(I)所示的肽，或其立体异构体、或其立体异构体的混合物，或其美容上可接受的盐、或其药学上可接受的盐，其特征在于，X 5是键，X 7是键。
  5. 根据权利要求1所述的式(I)所示的肽，或其立体异构体、或其立体异构体的混合物，或其美容上可接受的盐、或其药学上可接受的盐，其特征在于，选自下列肽(1)-(48)：
    (1)H-Val-Lys-Val-Leu-Gln-Trp-OH；
    (2)H-Val-Lys-Val-Leu-Gln-Trp-NH 2；
    (3)Ac-Val-Lys-Val-Leu-Gln-Trp-OH；
    (4)Ac-Val-Lys-Val-Leu-Gln-Trp-NH 2；
    (5)H-Ser-Met-Gly-Ala-Leu-Ala-OH；
    (6)H-Ser-Met-Gly-Ala-Leu-Ala-NH 2；
    (7)Ac-Ser-Met-Gly-Ala-Leu-Ala-OH；
    (8)Ac-Ser-Met-Gly-Ala-Leu-Ala-NH 2；
    (9)Ac-Val-His-Pro-Leu-Gln-Trp-OH；
    (10)Ac-Val-His-Pro-Leu-Gln-Trp-NH 2；
    (11)Ac-Ser-Arg-Leu-Pro-Leu-Trp-OH；
    (12)Ac-Ser-Arg-Leu-Pro-Leu-Trp-NH 2；
    (13)Ac-Trp-Lys-Val-Pro-Gln-Ala-OH；
    (14)Ac-Trp-Lys-Val-Pro-Gln-Ala-NH 2；
    (15)Ac-Val-Arg-Leu-Gly-Phe-Gln-His-Trp-OH；
    (16)Ac-Val-Arg-Leu-Gly-Phe-Gln-His-Trp-NH 2；
    (17)Ac-Val-His-Pro-Leu-Phe-Gln-His-Trp-OH；
    (18)Ac-Val-His-Pro-Leu-Phe-Gln-His-Trp-NH 2；
    (19)Ac-Val-Ala-Val-Leu-Phe-Gln-Arg-Trp-OH；
    (20)Ac-Val-Ala-Val-Leu-Phe-Gln-Arg-Trp-NH 2；
    (21)Ac-Val-Met-Gly-Pro-Trp-Leu-Lys-Ala-OH；
    (22)Ac-Val-Met-Gly-Pro-Trp-Leu-Lys-Ala-NH 2；
    (23)Ac-Val-Met-Ala-Pro-Tyr-Leu-Lys-Ala-OH；
    (24)Ac-Val-Met-Ala-Pro-Tyr-Leu-Lys-Ala-NH 2；
    (25)Ac-Ser-Ala-Val-Pro-Trp-Leu-Arg-Trp-OH；
    (26)Ac-Ser-Ala-Val-Pro-Trp-Leu-Arg-Trp-NH 2；
    (27)Ac-Ser-Met-Ala-Gly-Trp-Gln-His-Ala-OH；
    (28)Ac-Ser-Met-Ala-Gly-Trp-Gln-His-Ala-NH 2；
    (29)H-Trp-Ala-Ala-Pro-Phe-Leu-Arg-Ala-OH；
    (30)H-Trp-Ala-Ala-Pro-Phe-Leu-Arg-Ala-NH 2；
    (31)Ac-Trp-Ala-Ala-Pro-Phe-Leu-Arg-Ala-OH；
    (32)Ac-Trp-Ala-Ala-Pro-Phe-Leu-Arg-Ala-NH 2；
    (33)H-Trp-Lys-Ala-Leu-Phe-Gln-Arg-Trp-OH；
    (34)H-Trp-Lys-Gly-Pro-Tyr-Gln-Met-Trp-OH；
    (35)Ac-Met-Leu-Leu-Leu-Ala-Asp-NH 2；
    (36)Ac-Val-Asn-Gln-Leu-Val-Ala-NH 2；
    (37)Ac-Met-Leu-Asp-Phe-Leu-Ala-NH 2；
    (38)Ac-Leu-Trp-Phe-Val-Phe-Lys-NH 2；
    (39)Ac-Glu-Thr-Gly-Trp-Ala-Ala-NH 2；
    (40)Ac-Ala-Pro-Phe-Leu-Arg-Ala-NH 2；
    (41)Ac-Pro-Gly-Asp-Ala-Tyr-Gly-NH 2；
    (42)Ac-Tyr-Gly-Asp-Ala-Gly-Tyr-NH 2；
    (43)Ac-Ser-Phe-Ser-Asn-Gln-Met-NH 2；
    (44)Ac-Val-Asp-Cys-Ala-Asn-Lys-NH 2；
    (45)Ac-Val-Phe-Ala-Phe-Leu-Tyr-NH 2；
    (46)Ac-Ser-Ser-Ala-Trp-Ala-Lys-NH 2；
    (47)Ac-Thr-Cys-Lys-Glu-Leu-Arg-NH 2；
    (48)Ac-Leu-Asn-Leu-Val-Leu-Trp-NH 2。
  6. 根据权利要求5所述的肽，或其立体异构体、或其立体异构体的混合物，或其美容上可接受的盐、或其药学上可接受的盐，其特征在于，所述肽选自Ac-Val-Lys-Val-Leu-Gln-Trp-NH 2、Ac-Ser-Met-Gly-Ala-Leu-Ala-NH 2或Ac-Trp-Ala-Ala-Pro-Phe-Leu-Arg-Ala-NH 2。
  7. 根据权利要求1所述的式(I)所示的肽，或其立体异构体、或其立体异构体的混合物，或其美容上可接受的盐、或其药学上可接受的盐，其特征在于，
    所述美容上可接受的盐或药学上可接受的盐包括式(I)所示的肽的金属盐，其中，所述金属包括：锂、钠、钾、钙、镁、锰、铜、锌或铝；
    可选地，所述美容上可接受的盐或药学上可接受的盐包括式(I)所示的肽与有机碱形成的盐， 所述有机碱包括：乙胺、二乙胺、乙二胺、乙醇胺、二乙醇胺、精氨酸、赖氨酸、组氨酸或哌嗪；
    可选地，所述美容上可接受的盐或药学上可接受的盐包括式(I)所示的肽与无机酸或有机酸形成的盐，其中，所述有机酸包括：乙酸、柠檬酸、乳酸、丙二酸、马来酸、酒石酸、延胡索酸、苯甲酸、天冬氨酸、谷氨酸、琥珀酸、油酸、三氟乙酸、草酸、扑酸或葡萄糖酸；所述无机酸包括：盐酸、硫酸、硼酸或碳酸。
  8. 一种美容或药用组合物，其特征在于，包括有效量的如权利要求1-7任一项所述的式(I)所示的肽，或其立体异构体、或立体异构体的混合物，或其美容上可接受的盐、或其药学上可接受的盐，以及至少一种赋形剂和任选的美容上或药学上可接受的佐剂；
    可选地，所述佐剂选自：胶原合成刺激剂、调节PGC-1α合成的剂、调节PPARγ的活性的剂、增加或减少脂肪细胞的甘油三酸酯含量的剂、刺激或延迟脂肪细胞分化的剂、脂解剂或刺激脂肪分解的剂、溶脂剂、生脂剂、乙酰胆碱受体聚集的抑制剂、抑制肌肉收缩的剂、抗胆碱能试剂、弹性蛋白酶抑制剂、基质金属蛋白酶抑制剂、黑色素合成刺激或抑制剂、增白剂或脱色剂、促色素沉着剂、自晒黑剂、抗老化剂、NO-合酶抑制剂、5α-还原酶抑制剂、赖氨酰羟化酶和/或脯氨酰羟化酶的抑制剂、抗氧化剂、自由基清除剂和/或抗大气污染的剂、活性羰基类物质清除剂、抗糖化剂、抗组胺剂、抗病毒剂、抗寄生虫剂、乳化剂、润肤剂、有机溶剂、液体推进剂、皮肤调理剂、保湿剂、保留水分的物质、α羟基酸、β羟基酸、增湿剂、表皮水解酶、维生素、氨基酸、蛋白质、色素或着色剂、染料、生物聚合物、胶凝聚合物、增稠剂、表面活性剂、软化剂、粘合剂、防腐剂、抗皱剂、能够减少或治疗下眼袋的剂、去角质剂、角质剥离剂、角质层分离剂、抗微生物剂、抗真菌剂、抑真菌剂、灭菌剂、抑菌剂、刺激真皮或表皮大分子的合成和/或能够抑制或预防它们的降解的剂、刺激弹性蛋白合成的剂、刺激核心蛋白聚糖合成的剂、刺激层粘连蛋白合成的剂、刺激防御素合成的剂、刺激伴侣蛋白合成的剂、刺激cAMP合成的剂、热休克蛋白、刺激HSP70合成的剂、刺激热休克蛋白合成的剂、刺激透明质酸合成的剂、刺激纤连蛋白合成的剂、刺激去乙酰化酶合成的剂、刺激脂质和角质层组分的合成的剂、神经酰胺、脂肪酸、抑制胶原降解的剂、抑制弹性蛋白降解的剂、抑制丝氨酸蛋白酶的剂、刺激成纤维细胞增殖的剂、刺激角质形成细胞增殖的剂、刺激脂肪细胞增殖的剂、刺激黑色素细胞增殖的剂、刺激角质形成细胞分化的剂、抑制乙酰胆碱酯酶的剂、皮肤松弛剂、刺激糖胺聚糖合成的剂、抗角化过度剂、粉刺溶解剂、抗银屑病剂、抗皮炎剂、抗湿疹剂、DNA修复剂、DNA防护剂、稳定剂、止痒剂、用于治疗和/或护理敏感性皮肤的剂、固化剂、紧致剂、重构剂、抗拉伸纹剂、粘合剂、调节皮脂产生的剂、止汗剂、刺激愈合的剂、协助愈合的剂、刺激再上皮化的剂、协助再上皮化的剂、细胞因子生长因子、镇静剂、抗炎剂、麻醉剂、作用于毛细血管循环和/或微循环的剂、刺激血管生成的剂、抑制血管渗透性的剂、静脉紧张剂、作用于细胞代谢的剂、用于改善真皮-表皮接合的剂、诱导毛发生长的剂、毛发生长抑制或延缓剂、香料、螯合剂、植物提取物、精油、海洋提取物、得自生物发酵过程的剂、无机盐、细胞提取物、防晒剂、以及有效抗A和/或B紫外线的有机或无机光防护剂或其混合物。
  9. 根据权利要求8所述的美容或药用组合物，其特征在于，所述美容或药用组合物的制剂选自：霜剂、油、奶、香膏、泡沫、洗剂、凝胶、擦剂、浆液、皂、洗发精、润发乳、血清、软膏、摩丝、润发油、粉末、杆剂、笔剂、喷雾剂、气溶胶、胶囊剂、片剂、颗粒剂、口香糖、溶液、混悬液、乳剂、糖浆剂、酏剂、多糖薄膜、胶冻或明胶；
    可选地，所述胶囊剂包括：软胶囊剂、硬胶囊剂，可选为明胶胶囊剂；所述片剂包括：糖衣片剂。
  10. 一种美容上或药学上可接受的递送系统或缓释系统，其特征在于，包含有效量的如权利要求1-7任一项所述的式(I)所示的肽，或其立体异构体、或立体异构体的混合物，或其美容上可接受的盐、或其药学上可接受的盐，或权利要求8或9所述的美容或药用组合物；
    所述美容上或药学上可接受的递送系统或缓释系统选自：脂质体、油质体、非离子型表面活性剂脂质体囊泡、醇质体、毫米胶囊、微米胶囊、纳米胶囊、纳米结构的脂质载体、海绵状物、环糊精、类脂囊泡、胶束、毫米球、微米球、纳米球、脂质球、微米乳液、纳米乳液、毫米粒子、微米粒子或纳米粒子；可选为脂质体或微米乳液，可选具有反胶束的内部结构的油包水型微米乳液。
  11. 一种如权利要求1-7任一项所述的式(I)所示的肽，或其立体异构体、或立体异构体的混合物，或其美容上可接受的盐、或其药学上可接受的盐，或权利要求8或9所述美容或药用组合物，或权利要求10所述的美容上或药学上可接受的递送系统或缓释系统在制备用于治疗或护理由肌肉收缩导致的病况、失调或疾病的美容组合物或药用组合物中的用途，其中，所述失调或疾病是张力障碍。
  12. 一种如权利要求1-7任一项所述的式(I)所示的肽，或其立体异构体、或立体异构体的混合物，或其美容上可接受的盐、或其药学上可接受的盐，或权利要求8或9所述美容或药用组合物，或权利要求10所述的美容上或药学上可接受的递送系统或缓释系统在制备用于治疗、预防或修复皮肤老化或光老化的美容组合物或药用组合物中的用途，其中，所述皮肤老化或光老化的治疗、预防或修复是减少、预防或治疗面部皱纹。
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Cited By (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN117084934A (zh) * 2023-10-20 2023-11-21 深圳市维琪科技股份有限公司 多肽的新用途
CN117106021A (zh) * 2023-10-25 2023-11-24 深圳市维琪科技股份有限公司 六肽及其组合物和用途
CN117304265A (zh) * 2023-11-29 2023-12-29 深圳市维琪科技股份有限公司 一种皮肤护理多肽、组合物及其应用
CN117402216A (zh) * 2023-12-14 2024-01-16 深圳市维琪科技股份有限公司 八肽及其组合物和用途

Citations (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2010105636A1 (en) * 2009-03-17 2010-09-23 United Technologies Ut Ag Topical compositions comprising interleukin-1 alpha and peptides and cosmetic methods thereof

Family Cites Families (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US8946166B2 (en) * 2009-07-24 2015-02-03 Lipotec, S.A. Peptide-based compounds and compositions which inhibit muscle contraction
US20110053829A1 (en) * 2009-09-03 2011-03-03 Curevac Gmbh Disulfide-linked polyethyleneglycol/peptide conjugates for the transfection of nucleic acids
CN102675419A (zh) * 2011-03-16 2012-09-19 上海博智生物科技有限公司 一种Aβ短肽聚合抑制剂及其制备与应用
CN108042533B (zh) * 2017-12-25 2020-11-13 苏州格兰科医药科技有限公司 右美托米定在制备用于预防和/或抑制创伤性脑损伤后的神经元死亡的药物组合物中的用途
CN113896768B (zh) * 2021-11-01 2022-10-14 深圳市维琪医药研发有限公司 抗菌肽以及它们的美容组合物或药用组合物和用途

Patent Citations (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2010105636A1 (en) * 2009-03-17 2010-09-23 United Technologies Ut Ag Topical compositions comprising interleukin-1 alpha and peptides and cosmetic methods thereof

Cited By (8)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN117084934A (zh) * 2023-10-20 2023-11-21 深圳市维琪科技股份有限公司 多肽的新用途
CN117084934B (zh) * 2023-10-20 2024-01-19 深圳市维琪科技股份有限公司 多肽的新用途
CN117106021A (zh) * 2023-10-25 2023-11-24 深圳市维琪科技股份有限公司 六肽及其组合物和用途
CN117106021B (zh) * 2023-10-25 2023-12-26 深圳市维琪科技股份有限公司 六肽及其组合物和用途
CN117304265A (zh) * 2023-11-29 2023-12-29 深圳市维琪科技股份有限公司 一种皮肤护理多肽、组合物及其应用
CN117304265B (zh) * 2023-11-29 2024-01-30 深圳市维琪科技股份有限公司 一种皮肤护理多肽、组合物及其应用
CN117402216A (zh) * 2023-12-14 2024-01-16 深圳市维琪科技股份有限公司 八肽及其组合物和用途
CN117402216B (zh) * 2023-12-14 2024-02-13 深圳市维琪科技股份有限公司 八肽及其组合物和用途

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