CN107892718A - 一种多肽及其制备方法与应用 - Google Patents
一种多肽及其制备方法与应用 Download PDFInfo
- Publication number
- CN107892718A CN107892718A CN201711441042.9A CN201711441042A CN107892718A CN 107892718 A CN107892718 A CN 107892718A CN 201711441042 A CN201711441042 A CN 201711441042A CN 107892718 A CN107892718 A CN 107892718A
- Authority
- CN
- China
- Prior art keywords
- polypeptide
- skin
- preparation
- present
- amino acid
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Withdrawn
Links
- 108090000765 processed proteins & peptides Proteins 0.000 title claims abstract description 60
- 102000004196 processed proteins & peptides Human genes 0.000 title claims abstract description 42
- 229920001184 polypeptide Polymers 0.000 title claims abstract description 40
- 238000002360 preparation method Methods 0.000 title claims abstract description 19
- 230000006378 damage Effects 0.000 claims abstract description 13
- 230000000694 effects Effects 0.000 claims abstract description 12
- 239000003814 drug Substances 0.000 claims abstract description 6
- 230000036541 health Effects 0.000 claims abstract description 5
- 238000009098 adjuvant therapy Methods 0.000 claims abstract description 4
- 125000003275 alpha amino acid group Chemical group 0.000 claims abstract 3
- 125000003088 (fluoren-9-ylmethoxy)carbonyl group Chemical group 0.000 claims description 10
- 239000011347 resin Substances 0.000 claims description 10
- 229920005989 resin Polymers 0.000 claims description 10
- VYMPLPIFKRHAAC-UHFFFAOYSA-N 1,2-ethanedithiol Chemical compound SCCS VYMPLPIFKRHAAC-UHFFFAOYSA-N 0.000 claims description 5
- JGFZNNIVVJXRND-UHFFFAOYSA-N N,N-Diisopropylethylamine (DIPEA) Chemical compound CCN(C(C)C)C(C)C JGFZNNIVVJXRND-UHFFFAOYSA-N 0.000 claims description 5
- 238000005859 coupling reaction Methods 0.000 claims description 5
- 239000000243 solution Substances 0.000 claims description 5
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 5
- QGZKDVFQNNGYKY-UHFFFAOYSA-N Ammonia Chemical compound N QGZKDVFQNNGYKY-UHFFFAOYSA-N 0.000 claims description 4
- 239000003795 chemical substances by application Substances 0.000 claims description 4
- 230000008878 coupling Effects 0.000 claims description 4
- 238000010168 coupling process Methods 0.000 claims description 4
- 239000011259 mixed solution Substances 0.000 claims description 4
- BLRPTPMANUNPDV-UHFFFAOYSA-N Silane Chemical compound [SiH4] BLRPTPMANUNPDV-UHFFFAOYSA-N 0.000 claims description 3
- 210000004899 c-terminal region Anatomy 0.000 claims description 3
- 239000003153 chemical reaction reagent Substances 0.000 claims description 3
- 239000007822 coupling agent Substances 0.000 claims description 3
- 238000005336 cracking Methods 0.000 claims description 3
- 238000000354 decomposition reaction Methods 0.000 claims description 3
- 229910000077 silane Inorganic materials 0.000 claims description 3
- 239000007790 solid phase Substances 0.000 claims description 3
- 239000002253 acid Substances 0.000 claims description 2
- 229910021529 ammonia Inorganic materials 0.000 claims description 2
- 230000000903 blocking effect Effects 0.000 claims description 2
- CMWYAOXYQATXSI-UHFFFAOYSA-N n,n-dimethylformamide;piperidine Chemical compound CN(C)C=O.C1CCNCC1 CMWYAOXYQATXSI-UHFFFAOYSA-N 0.000 claims description 2
- 125000001449 isopropyl group Chemical group [H]C([H])([H])C([H])(*)C([H])([H])[H] 0.000 claims 1
- 208000028990 Skin injury Diseases 0.000 abstract description 3
- 238000012360 testing method Methods 0.000 abstract description 3
- 230000000192 social effect Effects 0.000 abstract description 2
- 238000010532 solid phase synthesis reaction Methods 0.000 abstract description 2
- 210000004027 cell Anatomy 0.000 description 37
- 210000003491 skin Anatomy 0.000 description 19
- 150000001413 amino acids Chemical group 0.000 description 16
- 230000004083 survival effect Effects 0.000 description 13
- YMWUJEATGCHHMB-UHFFFAOYSA-N Dichloromethane Chemical compound ClCCl YMWUJEATGCHHMB-UHFFFAOYSA-N 0.000 description 12
- 108010014258 Elastin Proteins 0.000 description 10
- 102000016942 Elastin Human genes 0.000 description 9
- 229920002549 elastin Polymers 0.000 description 9
- 239000000047 product Substances 0.000 description 8
- 238000001514 detection method Methods 0.000 description 7
- 239000007788 liquid Substances 0.000 description 7
- 238000011160 research Methods 0.000 description 7
- WFDIJRYMOXRFFG-UHFFFAOYSA-N Acetic anhydride Chemical compound CC(=O)OC(C)=O WFDIJRYMOXRFFG-UHFFFAOYSA-N 0.000 description 6
- IAZDPXIOMUYVGZ-UHFFFAOYSA-N Dimethylsulphoxide Chemical compound CS(C)=O IAZDPXIOMUYVGZ-UHFFFAOYSA-N 0.000 description 6
- ZMXDDKWLCZADIW-UHFFFAOYSA-N N,N-Dimethylformamide Chemical compound CN(C)C=O ZMXDDKWLCZADIW-UHFFFAOYSA-N 0.000 description 6
- JUJWROOIHBZHMG-UHFFFAOYSA-N Pyridine Chemical compound C1=CC=NC=C1 JUJWROOIHBZHMG-UHFFFAOYSA-N 0.000 description 6
- XLOMVQKBTHCTTD-UHFFFAOYSA-N Zinc monoxide Chemical compound [Zn]=O XLOMVQKBTHCTTD-UHFFFAOYSA-N 0.000 description 6
- 125000000539 amino acid group Chemical group 0.000 description 6
- 238000006243 chemical reaction Methods 0.000 description 6
- 238000005516 engineering process Methods 0.000 description 5
- 230000003834 intracellular effect Effects 0.000 description 5
- FEMOMIGRRWSMCU-UHFFFAOYSA-N ninhydrin Chemical compound C1=CC=C2C(=O)C(O)(O)C(=O)C2=C1 FEMOMIGRRWSMCU-UHFFFAOYSA-N 0.000 description 5
- IJGRMHOSHXDMSA-UHFFFAOYSA-N Atomic nitrogen Chemical compound N#N IJGRMHOSHXDMSA-UHFFFAOYSA-N 0.000 description 4
- OKKJLVBELUTLKV-UHFFFAOYSA-N Methanol Chemical compound OC OKKJLVBELUTLKV-UHFFFAOYSA-N 0.000 description 4
- GWEVSGVZZGPLCZ-UHFFFAOYSA-N Titan oxide Chemical compound O=[Ti]=O GWEVSGVZZGPLCZ-UHFFFAOYSA-N 0.000 description 4
- 230000032683 aging Effects 0.000 description 4
- 230000015572 biosynthetic process Effects 0.000 description 4
- 238000000034 method Methods 0.000 description 4
- 239000001301 oxygen Substances 0.000 description 4
- 229910052760 oxygen Inorganic materials 0.000 description 4
- 238000003786 synthesis reaction Methods 0.000 description 4
- WEVYAHXRMPXWCK-UHFFFAOYSA-N Acetonitrile Chemical compound CC#N WEVYAHXRMPXWCK-UHFFFAOYSA-N 0.000 description 3
- 241000251468 Actinopterygii Species 0.000 description 3
- DHMQDGOQFOQNFH-UHFFFAOYSA-N Glycine Chemical compound NCC(O)=O DHMQDGOQFOQNFH-UHFFFAOYSA-N 0.000 description 3
- 208000027418 Wounds and injury Diseases 0.000 description 3
- QVGXLLKOCUKJST-UHFFFAOYSA-N atomic oxygen Chemical compound [O] QVGXLLKOCUKJST-UHFFFAOYSA-N 0.000 description 3
- 239000000843 powder Substances 0.000 description 3
- UMJSCPRVCHMLSP-UHFFFAOYSA-N pyridine Natural products COC1=CC=CN=C1 UMJSCPRVCHMLSP-UHFFFAOYSA-N 0.000 description 3
- 239000003642 reactive oxygen metabolite Substances 0.000 description 3
- 239000011787 zinc oxide Substances 0.000 description 3
- UGNIYGNGCNXHTR-SFHVURJKSA-N (2s)-2-(9h-fluoren-9-ylmethoxycarbonylamino)-3-methylbutanoic acid Chemical compound C1=CC=C2C(COC(=O)N[C@@H](C(C)C)C(O)=O)C3=CC=CC=C3C2=C1 UGNIYGNGCNXHTR-SFHVURJKSA-N 0.000 description 2
- 108020004705 Codon Proteins 0.000 description 2
- RTZKZFJDLAIYFH-UHFFFAOYSA-N Diethyl ether Chemical compound CCOCC RTZKZFJDLAIYFH-UHFFFAOYSA-N 0.000 description 2
- ROHFNLRQFUQHCH-YFKPBYRVSA-N L-leucine Chemical compound CC(C)C[C@H](N)C(O)=O ROHFNLRQFUQHCH-YFKPBYRVSA-N 0.000 description 2
- AYFVYJQAPQTCCC-GBXIJSLDSA-N L-threonine Chemical compound C[C@@H](O)[C@H](N)C(O)=O AYFVYJQAPQTCCC-GBXIJSLDSA-N 0.000 description 2
- CBENFWSGALASAD-UHFFFAOYSA-N Ozone Chemical compound [O-][O+]=O CBENFWSGALASAD-UHFFFAOYSA-N 0.000 description 2
- 238000003556 assay Methods 0.000 description 2
- 230000008859 change Effects 0.000 description 2
- 239000012043 crude product Substances 0.000 description 2
- 238000005520 cutting process Methods 0.000 description 2
- 201000010099 disease Diseases 0.000 description 2
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 2
- 239000000839 emulsion Substances 0.000 description 2
- 238000004128 high performance liquid chromatography Methods 0.000 description 2
- 208000014674 injury Diseases 0.000 description 2
- 238000001294 liquid chromatography-tandem mass spectrometry Methods 0.000 description 2
- 229910052757 nitrogen Inorganic materials 0.000 description 2
- 239000000516 sunscreening agent Substances 0.000 description 2
- 239000004408 titanium dioxide Substances 0.000 description 2
- 238000012549 training Methods 0.000 description 2
- ZGYICYBLPGRURT-UHFFFAOYSA-N tri(propan-2-yl)silicon Chemical compound CC(C)[Si](C(C)C)C(C)C ZGYICYBLPGRURT-UHFFFAOYSA-N 0.000 description 2
- 210000004881 tumor cell Anatomy 0.000 description 2
- DWNBOPVKNPVNQG-LURJTMIESA-N (2s)-4-hydroxy-2-(propylamino)butanoic acid Chemical compound CCCN[C@H](C(O)=O)CCO DWNBOPVKNPVNQG-LURJTMIESA-N 0.000 description 1
- OYULCCKKLJPNPU-DIFFPNOSSA-N (2s,3r)-2-(9h-fluoren-9-ylmethoxycarbonylamino)-3-hydroxybutanoic acid Chemical compound C1=CC=C2C(COC(=O)N[C@@H]([C@H](O)C)C(O)=O)C3=CC=CC=C3C2=C1 OYULCCKKLJPNPU-DIFFPNOSSA-N 0.000 description 1
- NDKDFTQNXLHCGO-UHFFFAOYSA-N 2-(9h-fluoren-9-ylmethoxycarbonylamino)acetic acid Chemical compound C1=CC=C2C(COC(=O)NCC(=O)O)C3=CC=CC=C3C2=C1 NDKDFTQNXLHCGO-UHFFFAOYSA-N 0.000 description 1
- RLCSROTYKMPBDL-USJZOSNVSA-N 2-[[(2s)-1-[(2s)-2-[[(2s)-2-[[2-[[(2s)-2-amino-3-methylbutanoyl]amino]acetyl]amino]-3-methylbutanoyl]amino]propanoyl]pyrrolidine-2-carbonyl]amino]acetic acid Chemical compound CC(C)[C@H](N)C(=O)NCC(=O)N[C@@H](C(C)C)C(=O)N[C@@H](C)C(=O)N1CCC[C@H]1C(=O)NCC(O)=O RLCSROTYKMPBDL-USJZOSNVSA-N 0.000 description 1
- VAJVDSVGBWFCLW-UHFFFAOYSA-N 3-Phenyl-1-propanol Chemical compound OCCCC1=CC=CC=C1 VAJVDSVGBWFCLW-UHFFFAOYSA-N 0.000 description 1
- 102100026189 Beta-galactosidase Human genes 0.000 description 1
- 102000008186 Collagen Human genes 0.000 description 1
- 108010035532 Collagen Proteins 0.000 description 1
- 102100033167 Elastin Human genes 0.000 description 1
- 102000004190 Enzymes Human genes 0.000 description 1
- 108090000790 Enzymes Proteins 0.000 description 1
- 239000004471 Glycine Substances 0.000 description 1
- FFEARJCKVFRZRR-BYPYZUCNSA-N L-methionine Chemical compound CSCC[C@H](N)C(O)=O FFEARJCKVFRZRR-BYPYZUCNSA-N 0.000 description 1
- ROHFNLRQFUQHCH-UHFFFAOYSA-N Leucine Natural products CC(C)CC(N)C(O)=O ROHFNLRQFUQHCH-UHFFFAOYSA-N 0.000 description 1
- 206010027146 Melanoderma Diseases 0.000 description 1
- 229930182555 Penicillin Natural products 0.000 description 1
- JGSARLDLIJGVTE-MBNYWOFBSA-N Penicillin G Chemical compound N([C@H]1[C@H]2SC([C@@H](N2C1=O)C(O)=O)(C)C)C(=O)CC1=CC=CC=C1 JGSARLDLIJGVTE-MBNYWOFBSA-N 0.000 description 1
- DYUQAZSOFZSPHD-UHFFFAOYSA-N Phenylpropyl alcohol Natural products CCC(O)C1=CC=CC=C1 DYUQAZSOFZSPHD-UHFFFAOYSA-N 0.000 description 1
- 208000012641 Pigmentation disease Diseases 0.000 description 1
- 241000269851 Sarda sarda Species 0.000 description 1
- 206010043189 Telangiectasia Diseases 0.000 description 1
- AYFVYJQAPQTCCC-UHFFFAOYSA-N Threonine Natural products CC(O)C(N)C(O)=O AYFVYJQAPQTCCC-UHFFFAOYSA-N 0.000 description 1
- 239000004473 Threonine Substances 0.000 description 1
- 230000037338 UVA radiation Effects 0.000 description 1
- 238000002835 absorbance Methods 0.000 description 1
- 150000008065 acid anhydrides Chemical class 0.000 description 1
- 239000004480 active ingredient Substances 0.000 description 1
- 239000011149 active material Substances 0.000 description 1
- 230000002776 aggregation Effects 0.000 description 1
- 238000004220 aggregation Methods 0.000 description 1
- 239000005030 aluminium foil Substances 0.000 description 1
- 230000003064 anti-oxidating effect Effects 0.000 description 1
- XNEFYCZVKIDDMS-UHFFFAOYSA-N avobenzone Chemical compound C1=CC(OC)=CC=C1C(=O)CC(=O)C1=CC=C(C(C)(C)C)C=C1 XNEFYCZVKIDDMS-UHFFFAOYSA-N 0.000 description 1
- 229960005193 avobenzone Drugs 0.000 description 1
- 230000008827 biological function Effects 0.000 description 1
- 201000011510 cancer Diseases 0.000 description 1
- 239000002775 capsule Substances 0.000 description 1
- 238000004113 cell culture Methods 0.000 description 1
- 239000012930 cell culture fluid Substances 0.000 description 1
- 239000006143 cell culture medium Substances 0.000 description 1
- 230000022131 cell cycle Effects 0.000 description 1
- 230000008809 cell oxidative stress Effects 0.000 description 1
- 230000001413 cellular effect Effects 0.000 description 1
- 238000005119 centrifugation Methods 0.000 description 1
- -1 chlorazol tetramethyl hexafluorophosphates Chemical class 0.000 description 1
- 229920001436 collagen Polymers 0.000 description 1
- 238000004737 colorimetric analysis Methods 0.000 description 1
- 230000009514 concussion Effects 0.000 description 1
- 238000010276 construction Methods 0.000 description 1
- 239000006071 cream Substances 0.000 description 1
- 238000013461 design Methods 0.000 description 1
- 238000010586 diagram Methods 0.000 description 1
- 229940079593 drug Drugs 0.000 description 1
- 238000004043 dyeing Methods 0.000 description 1
- 108010064033 elastin-binding proteins Proteins 0.000 description 1
- 229920001971 elastomer Polymers 0.000 description 1
- 239000000806 elastomer Substances 0.000 description 1
- 239000003480 eluent Substances 0.000 description 1
- 238000010828 elution Methods 0.000 description 1
- 230000002708 enhancing effect Effects 0.000 description 1
- UVCJGUGAGLDPAA-UHFFFAOYSA-N ensulizole Chemical compound N1C2=CC(S(=O)(=O)O)=CC=C2N=C1C1=CC=CC=C1 UVCJGUGAGLDPAA-UHFFFAOYSA-N 0.000 description 1
- 229960000655 ensulizole Drugs 0.000 description 1
- 210000001339 epidermal cell Anatomy 0.000 description 1
- 210000002615 epidermis Anatomy 0.000 description 1
- 230000005284 excitation Effects 0.000 description 1
- 239000004744 fabric Substances 0.000 description 1
- 230000001815 facial effect Effects 0.000 description 1
- 239000000835 fiber Substances 0.000 description 1
- 239000000706 filtrate Substances 0.000 description 1
- 238000009472 formulation Methods 0.000 description 1
- 230000006870 function Effects 0.000 description 1
- 239000001963 growth medium Substances 0.000 description 1
- 238000000338 in vitro Methods 0.000 description 1
- 230000009545 invasion Effects 0.000 description 1
- 239000007791 liquid phase Substances 0.000 description 1
- 238000004519 manufacturing process Methods 0.000 description 1
- 239000000463 material Substances 0.000 description 1
- 229930182817 methionine Natural products 0.000 description 1
- 239000000203 mixture Substances 0.000 description 1
- 210000002569 neuron Anatomy 0.000 description 1
- 239000002773 nucleotide Substances 0.000 description 1
- 125000003729 nucleotide group Chemical group 0.000 description 1
- 235000015097 nutrients Nutrition 0.000 description 1
- FMJSMJQBSVNSBF-UHFFFAOYSA-N octocrylene Chemical group C=1C=CC=CC=1C(=C(C#N)C(=O)OCC(CC)CCCC)C1=CC=CC=C1 FMJSMJQBSVNSBF-UHFFFAOYSA-N 0.000 description 1
- 229960000601 octocrylene Drugs 0.000 description 1
- 230000036542 oxidative stress Effects 0.000 description 1
- 229940049954 penicillin Drugs 0.000 description 1
- 238000010647 peptide synthesis reaction Methods 0.000 description 1
- 239000012071 phase Substances 0.000 description 1
- 230000019612 pigmentation Effects 0.000 description 1
- 150000003053 piperidines Chemical class 0.000 description 1
- 238000002953 preparative HPLC Methods 0.000 description 1
- 230000008569 process Effects 0.000 description 1
- 238000012545 processing Methods 0.000 description 1
- 102000004169 proteins and genes Human genes 0.000 description 1
- 108090000623 proteins and genes Proteins 0.000 description 1
- 238000000746 purification Methods 0.000 description 1
- 230000005855 radiation Effects 0.000 description 1
- 230000035484 reaction time Effects 0.000 description 1
- 230000009758 senescence Effects 0.000 description 1
- 210000002966 serum Anatomy 0.000 description 1
- 239000012679 serum free medium Substances 0.000 description 1
- 230000037380 skin damage Effects 0.000 description 1
- 239000007787 solid Substances 0.000 description 1
- 238000001228 spectrum Methods 0.000 description 1
- 206010041823 squamous cell carcinoma Diseases 0.000 description 1
- 239000000126 substance Substances 0.000 description 1
- 230000000475 sunscreen effect Effects 0.000 description 1
- 239000006228 supernatant Substances 0.000 description 1
- 230000001629 suppression Effects 0.000 description 1
- 230000008961 swelling Effects 0.000 description 1
- 238000004885 tandem mass spectrometry Methods 0.000 description 1
- 208000009056 telangiectasis Diseases 0.000 description 1
- 238000009281 ultraviolet germicidal irradiation Methods 0.000 description 1
- 108010011876 valyl-glycyl-valyl-alanyl-prolyl-glycine Proteins 0.000 description 1
- 230000037303 wrinkles Effects 0.000 description 1
Classifications
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
- C07K14/435—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
- C07K14/78—Connective tissue peptides, e.g. collagen, elastin, laminin, fibronectin, vitronectin or cold insoluble globulin [CIG]
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K8/00—Cosmetics or similar toiletry preparations
- A61K8/18—Cosmetics or similar toiletry preparations characterised by the composition
- A61K8/30—Cosmetics or similar toiletry preparations characterised by the composition containing organic compounds
- A61K8/64—Proteins; Peptides; Derivatives or degradation products thereof
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61Q—SPECIFIC USE OF COSMETICS OR SIMILAR TOILETRY PREPARATIONS
- A61Q17/00—Barrier preparations; Preparations brought into direct contact with the skin for affording protection against external influences, e.g. sunlight, X-rays or other harmful rays, corrosive materials, bacteria or insect stings
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K38/00—Medicinal preparations containing peptides
Landscapes
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- General Health & Medical Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Public Health (AREA)
- Animal Behavior & Ethology (AREA)
- Veterinary Medicine (AREA)
- Organic Chemistry (AREA)
- Gastroenterology & Hepatology (AREA)
- Molecular Biology (AREA)
- Dermatology (AREA)
- Biochemistry (AREA)
- Biophysics (AREA)
- Genetics & Genomics (AREA)
- Medicinal Chemistry (AREA)
- Zoology (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- Toxicology (AREA)
- Birds (AREA)
- Epidemiology (AREA)
- Medicines That Contain Protein Lipid Enzymes And Other Medicines (AREA)
- Peptides Or Proteins (AREA)
Abstract
本发明属于生物制药领域,具体涉及一种多肽及其制备方法与应用。本发明所述多肽其氨基酸序列如SEQ ID NO:1所示。本发明所述多肽使用多肽合成仪,采用固相合成法合成。实验结果表明本发明所述多肽具有很好的具有抗皮肤UV损伤活性,可以用于制备皮肤损伤的辅助治疗药物或保健品以及用于防止皮肤紫外损伤的护肤品,应用范围广泛,具有良好的经济和社会效应。
Description
技术领域
本发明属于生物制药领域,具体涉及一种多肽及其制备方法与应用,尤其是一种具有抗皮肤UV损伤活性的多肽及其制备方法与应用。
背景技术
研究表明皮肤衰老的元凶并不是自然老化,更多的是由于光老化引起的。光老化是由于皮肤长期受到日光照射所引起的损害,表现为皮肤粗糙、增厚、松弛、深而粗的皱纹,局部有过度的色素沉着或毛细血管扩张,甚至可能出现各种良性或恶性肿瘤(如日光角化病、鳞状细胞癌、恶性黑素瘤等)。日光中,紫外线的分类有UVA、UVB、UVC。其中UVC因为波长较短,在大气中就已经被臭氧层给吸收、散射,无法到达地面。UVB的波长居三者之中,波长仅能达到肌肤的表皮。而波长较长的UVA,波长320~420nm,又称为长波黑斑效应紫外线,超过98%能穿透臭氧层和云层到达地球表面,会深入肌肤的深层,伤害肌肤的真皮层,破坏弹性纤维和胶原蛋白纤维而造成皮肤晒黑、肌肤老化。因此,长波紫外线UVA在皮肤光老化过程中起着主要作用。
目前,女性的护肤品和防晒霜产品研究多围绕抑制UVA损伤进行活性物质研究。物理防晒的有效成分只有两种,Zinc Oxide(氧化锌)和Titanium Dioxide(二氧化钛),ZincOxide是一种非常好的防晒成分,其物理性粉体能反射散射紫外线UVA、UVB,安全度高,超过5%的高浓度时起效。化学防晒主要有:Avobenzone、MexorylSX、Octocrylene和Ensulizole等。
弹性蛋白肽是弹性蛋白原的降解产物,在维持人体细胞的生长和弹性起着非常重要的作用。自1982年开始已有弹性蛋白肽在皮肤科领域的研究,多年研究发现,弹性蛋白肽通过作用于细胞表面地受体弹性蛋白结合蛋白而发挥多种生物学功能,如促进正常细胞和肿瘤细胞蛋白酶的释放、增强肿瘤细胞的侵袭性和促进皮肤损伤的修复等。而尚未报道弹性蛋白肽在皮肤光老化和真皮弹性纤维疾病中是否具有作用。目前,弹性蛋白肽的研究已有六肽VGVAPG和九肽GPGVGAGVP、GLGBGAGVP等。而弹性蛋白七肽的研究尚未见报道。
发明内容
有鉴于此,本发明提供了一种对神经细胞具有抗氧化作用以及抑制Aβ42聚集的多肽。
为实现本发明的目的,本发明采用如下技术方案。
本发明从鲣鱼弹性蛋白肽来源,通过LC-MS/MS技术鉴定获得的弹性蛋白七肽,其氨基酸序列如SEQ ID NO:1所示。
具体的,本发明所述的多肽为:Thr-Gly-Val-Leu-Thr-Val-Met或者TGVLTVM,其中Thr为苏氨酸的氨基酸残基,Gly为甘氨酸的氨基酸残基,Val为颉氨酸的氨基酸残基,Leu为亮氨酸的氨基酸残基,Met为甲硫氨酸的氨基酸残基。
本发明也提供了编码本发明所述多肽的DNA分子。由于密码子的简并性,可以存在很多种能够编码本发明所述的特定多肽的核苷酸序列。对于编码本发明所述多肽的DNA分子,本领域技术人员可以很容易的利用现有公知的方法制造合成。诸如,通过选择对应于构成所述的氨基酸序列的氨基酸残基的密码子,可很容易地确定和提供相应于所述多肽的氨基酸序列的DNA分子。
本发明所述多肽可以采用成熟的多肽合成技术Fmoc方案合成,纯度>99%。
本发明提供了所述多肽的制备方法,采用标准的Fmoc方案,选用2-氯三苯基氯树脂树脂进行固相合成。
具体的,所述多肽的制备方法为以2-氯三苯基氯树脂为固相载体,按照SEQ IDNO:1所示氨基酸序列采用逐一偶联的方式从C端向N端偶联Fmoc保护基团的氨基酸得到肽树脂,裂解后得到多肽。。
其中,所述逐一偶联的偶联剂优选为HBTU/DIEA。所述偶联剂用量优选为过量。
进一步的,所述逐一偶联方式中在每步偶联前还包括脱除Fmoc步骤。在一些实施例中,所述脱除Fmoc的试剂为20%哌啶DMF溶液,即哌啶:DMF=1:4(体积比),反应时间为20min。
本发明所述多肽的制备方法中所述裂解的裂解剂优选为TFA、乙二硫醇、三异丙基硅烷与水的混合溶液。更优选为TFA、乙二硫醇、三异丙基硅烷与水的体积比为95:2:2:1的混合溶液。
本发明所述制备方法还包括对制得的多肽进行纯化的步骤。优选采用高效液相色谱提纯多肽。
在一些实施方案中,本发明采用UV光损伤Hacat人永生化表皮细胞模型,利用MTT和流式细胞仪检测细胞周期技术在体外细胞水平鉴定了本发明所述多肽的抗UV损伤活性。结果显示孵育本发明多肽后细胞在高剂量紫外UVA照射损伤下的存活率提高了20%,细胞周期G2期升高,Hacat细胞呈现出旺盛的生长、增值状态。表明本发明所述多肽具有抗UV损伤活性。
因此本发明提供了所述多肽在制备具有抗皮肤UV损伤活性的产品中的应用。
其中,所述具有抗皮肤UV损伤活性的产品为皮肤损伤的辅助治疗药物或保健品、用于防止皮肤紫外损伤的护肤品。
本领域技术人员可将本发明所述多肽直接或间接加入制备药物、保健品或护肤品时所需的各种常用辅料,以常规制备方法,制成常用剂型,如粉剂(胶囊、冻干粉)、乳剂、霜剂、凝胶、医用敷料等药物或保健品制剂,以及精华液、乳液、面霜、面膜等护肤品。
由上述技术方案可知,本发明提供了一种多肽及其制备方法与应用。本发明所述多肽其氨基酸序列如SEQ ID NO:1所示。本发明所述多肽使用多肽合成仪,采用固相合成法合成。实验结果表明本发明所述多肽具有很好的具有抗皮肤UV损伤活性,可以用于制备皮肤损伤的辅助治疗药物或保健品以及用于防止皮肤紫外损伤的护肤品,应用范围广泛,具有良好的经济和社会效应。
附图说明
为了更清楚地说明本发明实施例或现有技术中的技术方案,下面将对实施例或现有技术描述中所需要使用的附图作简单地介绍。
图1为MTT测细胞存活率柱状图;
图2为流式细胞仪测细胞内活性氧含量图。
具体实施方式
本发明公开了一种多肽及其制备方法与应用。本领域技术人员可以借鉴本文内容,适当改进工艺参数实现。特别需要指出的是,所有类似的替换和改动对本领域技术人员来说是显而易见的,它们都被视为包括在本发明。本发明的方法及产品已经通过较佳实施例进行了描述,相关人员明显能在不脱离本发明内容、精神和范围内对本文所述的方法进行改动或适当变更与组合,来实现和应用本发明技术。
为了进一步理解本发明,下面将结合本发明实施例,对本发明实施例中的技术方案进行清楚、完整地描述,显然,所描述的实施例仅仅是本发明一部分实施例,而不是全部的实施例。基于本发明中的实施例,本领域普通技术人员在没有做出创造性劳动前提下所获得的所有其他实施例,都属于本发明保护的范围。
如无特殊说明,本发明实施例中所涉及的试剂均为市售产品,均可以通过商业渠道购买获得。其中通过LC-MS/MS技术鉴定获得本发明所述多肽的具体参数为C18柱进行HPLC,进样量:20μl,流速:0.2ml/min,洗脱液:乙腈,梯度洗脱,MS/MS检索50-1000m/z,运用mascot软件搜库,搜库数据库Swissprot,物种选择Actinopterygii(ray-finned fish)辐鳍鱼。
实施例1、固相合成多肽
称取n当量2-氯三苯基氯树脂加入反应器,加入DCM(二氯甲烷)溶胀半小时,然后抽掉DCM,加入序列中第一个氨基酸Fmoc-Thr(tbu)-OH2n当量,加2n当量的DIEA和适量(使树脂充分鼓动后停止加入)的DMF、DCM,氮气鼓泡反应60min。然后加入5n当量甲醇,反应半小时,抽掉反应液,用DMF、MEOH洗净。往反应器中加入序列中第二个氨基酸Fmoc-Gly-OH,2n当量HBTU(1-羟基,苯丙,三氯唑四甲基六氟磷酸盐)及DIEA,N2鼓泡反应半小时,洗掉液体,茚三酮检测,然后用吡啶和乙酸酐封端。最后洗净,加入适量的脱帽液去除Fmoc(9-芴甲氧羰基)保护基,洗净,茚三酮检测。按氨基酸序列Thr-Gly-Val-Leu-Thr-Val-Met的序列特征,使肽链从C端逐个向N端延伸,依次加入第三个氨基酸Fmoc-Val-OH、第四个氨基酸Fmoc-Leu-OH、第五个氨基酸Fmoc-Thr-OH、第六个氨基酸Fmoc-Val-OH和第七个氨基酸Fmoc-Met-OH。每加入一个氨基酸,重复上述操作:加入2n当量HBTU及DIEA,N2鼓泡反应半小时,洗掉液体,茚三酮检测。然后用吡啶和乙酸酐封端。最后洗净,加入适量的脱帽液去除Fmoc(9-芴甲氧羰基)保护基,洗净,茚三酮检测。至加入第七个氨基酸后,将茚三酮检测后,用吡啶和乙酸酐封端。至最后一个氨基酸完成耦合反应,树脂用氮气吹干后从反应柱中取下,倒入烧瓶中,然后往烧瓶中加切割液与树脂比为10mL/克比例的切割液(95%TFA,2%乙二硫醇,2%三异丙基硅烷,1%水),震荡,滤掉树脂。往滤液中加入大量乙醚。析出粗产物,离心,清洗得多肽粗产物。用制备型高效液相色谱提纯粗产物,冻干为白色粉末。纯品经HPLC鉴定浓度为98.66%。
实施例2、构造皮肤光老化模型
用含10%胎牛血清,100U/mL青霉素、链霉素的MEM培养基对HaCaT细胞进行传代培养。取对数生长期的HaCaT细胞,细胞密度为3×104个/mL接种到96孔培养板,每孔100μL。培养24h后,吸出含血清的培养液,用D-Hanks液漂洗1次,每孔加入100μL的PBS接受UVA照射。UVA辐射仪光源离培养板的高度为20cm,垂直照射,经辐照度监测仪测UVA照射剂量为3J/cm2。为避免照射过程中培养基温度升高对细胞可能造成的伤害,将培养板底部置垫冰袋。照射后立即换新鲜培养基,置培养箱继续孵育16h后进行MTT比色法测定细胞存活率。假照射(对照组)组用铝箔纸完全遮盖避光。每处理组设5个复孔。
MTT法检测上述步骤处理后细胞存活率。将96孔板中接受UVA照射后的Hacat细胞置于37℃、5%CO2培养箱中继续培养12h,然后向每100μL培养基中加入5g/L MTT 10μL,继续孵育4h后,弃去上清,每孔加入二甲基亚砜(DMSO)100μL,室温下振荡15min,酶标仪于492nm波长处测定各孔吸光度(A)值,按照以下公式计算细胞存活率:细胞存活率=样品组OD490nm/对照组OD490nm×100%。MTT法测UV造模细胞存活率,结果如图1。
图1显示,3J/cm2UV剂量辐照Hacat细胞造模,检测空白对照(造模细胞未孵育肽)和孵育0.5mM、0.05mM细胞的存活率,MTT检测OD490nm发现,0.5mM肽孵育细胞存活率为125.6%,对比造模细胞存活率提高25.6%。0.05mM肽孵育细胞存活率为124.6%,对比造模细胞存活率提高24.6%。孵育肽细胞UV辐照后存活率显著性提高。
流式细胞仪检测细胞内活性氧含量变化。按照1:1000用无血清培养液稀释DCFH-DA,使终浓度为10微摩尔/升。去除细胞培养液,加入适当体积稀释好的DCFH-DA。加入的体积以能充分盖住细胞为宜,通常对于六孔板的一个孔加入稀释好的DCFH-DA不少于1毫升。37℃细胞培养箱内孵育20分钟。用无血清细胞培养液洗涤细胞三次,以充分去除未进入细胞内的DCFH-DA。收集细胞后用流式细胞仪488nm激发波长,525nm发射波长检测。流式细胞仪检测细胞内活性氧含量,结果如图2。
3J/cm2UV剂量辐照Hacat细胞造模,试剂盒ROS染色后,流式细胞仪检测空白对照(不造模细胞)、造模细胞和0.5mM肽孵育后造模细胞的胞内活性氧ROS含量变化。从流式细胞仪检测结果发现:对比空白对照和造模组,造模后细胞内活性氧显著性提高,造成细胞的氧化应激损伤,造模成功。对比造模组(未孵育肽)和孵育肽细胞内ROS,发现孵育肽细胞内活性氧荧光强度X-mean显著性降低,阳性细胞百分数显著性下降,两方面数据表明孵育肽hacat细胞内的活性氧含量显著性降低,UV造模造成的氧化应激损伤减弱,UV对细胞的损伤降低。
序列表
<110> 无限极(中国)有限公司
<120> 一种多肽及其制备方法与应用
<130> MP1719396
<160> 1
<170> SIPOSequenceListing 1.0
<210> 1
<211> 7
<212> PRT
<213> 鲣鱼弹性蛋白(Bonito elastin)
<400> 1
Thr Gly Val Leu Thr Val Met
1 5
Claims (9)
1.一种多肽,其特征在于,其氨基酸序列如SEQ ID NO:1所示。
2.编码权利要求1所述的多肽的DNA分子。
3.权利要求1所述的多肽的制备方法,其特征在于,以2-氯三苯基氯树脂为固相载体,按照SEQ ID NO:1所示氨基酸序列采用逐一偶联的方式从C端向N端偶联Fmoc保护基团的氨基酸得到肽树脂,裂解后得到多肽。
4.根据权利要求3所述制备方法,其特征在于,所述逐一偶联的偶联剂为HBTU/DIEA。
5.根据权利要求3所述制备方法,其特征在于,所述逐一偶联方式中在每步偶联前还包括脱除Fmoc步骤;所述脱除Fmoc的试剂为20%哌啶DMF溶液。
6.根据权利要求3所述制备方法,其特征在于,所述裂解的裂解剂为TFA、乙二硫醇、三异丙基硅烷与水的混合溶液。
7.根据权利要求3所述制备方法,其特征在于,所述裂解剂为TFA、乙二硫醇、三异丙基硅烷与水的体积比为95:2:2:1的混合溶液。
8.权利要求1所述多肽在制备具有抗皮肤UV损伤活性的产品中的应用。
9.根据权利要求8所述应用,其特征在于,所述具有抗皮肤UV损伤活性的产品为皮肤损伤的辅助治疗药物或保健品、用于防止皮肤紫外损伤的护肤品。
Priority Applications (1)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
CN201711441042.9A CN107892718A (zh) | 2017-12-27 | 2017-12-27 | 一种多肽及其制备方法与应用 |
Applications Claiming Priority (1)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
CN201711441042.9A CN107892718A (zh) | 2017-12-27 | 2017-12-27 | 一种多肽及其制备方法与应用 |
Publications (1)
Publication Number | Publication Date |
---|---|
CN107892718A true CN107892718A (zh) | 2018-04-10 |
Family
ID=61808799
Family Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
CN201711441042.9A Withdrawn CN107892718A (zh) | 2017-12-27 | 2017-12-27 | 一种多肽及其制备方法与应用 |
Country Status (1)
Country | Link |
---|---|
CN (1) | CN107892718A (zh) |
Cited By (2)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN112480209A (zh) * | 2020-11-16 | 2021-03-12 | 昆明学院 | 一种抗皮肤光损伤保护活性多肽rl-pl9及其应用 |
CN113683662A (zh) * | 2021-03-05 | 2021-11-23 | 四川大学 | 一种防护紫外皮肤损伤的多肽 |
Citations (3)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US20130338221A1 (en) * | 2012-06-19 | 2013-12-19 | Pukyong National University Industry-University Cooperation Foundation | Photoaging protective composition containing chromenes derived from sargassum horneri |
CN105218639A (zh) * | 2015-09-28 | 2016-01-06 | 华南理工大学 | 一种七肽及其应用 |
WO2017216177A1 (en) * | 2016-06-14 | 2017-12-21 | Sederma | Peptide, composition comprising said peptide and uses thereof, in particular cosmetic uses |
-
2017
- 2017-12-27 CN CN201711441042.9A patent/CN107892718A/zh not_active Withdrawn
Patent Citations (3)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US20130338221A1 (en) * | 2012-06-19 | 2013-12-19 | Pukyong National University Industry-University Cooperation Foundation | Photoaging protective composition containing chromenes derived from sargassum horneri |
CN105218639A (zh) * | 2015-09-28 | 2016-01-06 | 华南理工大学 | 一种七肽及其应用 |
WO2017216177A1 (en) * | 2016-06-14 | 2017-12-21 | Sederma | Peptide, composition comprising said peptide and uses thereof, in particular cosmetic uses |
Non-Patent Citations (1)
Title |
---|
田珊等: "弹性蛋白肽在皮肤科应用研究的进展", 《中国皮肤性病学杂志》 * |
Cited By (2)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN112480209A (zh) * | 2020-11-16 | 2021-03-12 | 昆明学院 | 一种抗皮肤光损伤保护活性多肽rl-pl9及其应用 |
CN113683662A (zh) * | 2021-03-05 | 2021-11-23 | 四川大学 | 一种防护紫外皮肤损伤的多肽 |
Similar Documents
Publication | Publication Date | Title |
---|---|---|
CN100590130C (zh) | 抗癌活性肽 | |
CN104023737B (zh) | 抑制被激活受体的肽及其在化妆品组合物或药物组合物中的用途 | |
CN104968360A (zh) | 用于治疗和/或护理皮肤、毛发和/或粘膜的化合物以及它们的化妆美容或药物组合物 | |
CN101687935B (zh) | Tgfp-cap肽及其用途 | |
KR101508614B1 (ko) | 스파이더 실크 단백질, 거머리 추출물, 봉독 추출물 및 뱀독 유사 펩타이드 복합물로 제조된 포이즌 컴플렉스를 유효성분으로 함유하는 항노화 화장료 조성물 | |
CN113248573B (zh) | 改善皮肤生理特性的活性肽与间充质干细胞外泌体在药品或化妆品中的应用 | |
CN108495646A (zh) | 用于治疗和/或护理皮肤、毛发、指甲和/或粘膜的化合物 | |
CN105131086A (zh) | 一种六肽及其应用 | |
CN107619437A (zh) | 一种皮肤创伤修复肽whpp‑oa1及其提纯方法与应用 | |
CN106943338A (zh) | 个性化美容护肤品及其制备方法 | |
CN112771064A (zh) | 可用于对皮肤、毛发、指甲和/或粘膜进行处理和/或护理的化合物 | |
CN107892718A (zh) | 一种多肽及其制备方法与应用 | |
CN116284241A (zh) | 新的肽及其组合物和用途 | |
CN105218639A (zh) | 一种七肽及其应用 | |
CN101679495B (zh) | 用于细胞学和免疫学调节的短的生物活性肽 | |
CN110590912A (zh) | 一种新型抗氧化活性多肽oa-vi12及其制备方法与应用 | |
CN116535463B (zh) | 活性肽及其组合物和用途 | |
CN107955062B (zh) | 一种合成多肽及其合成方法和应用 | |
KR102130988B1 (ko) | 꿀, 회화나무, 동아씨, 오미자, 괄루근, 상백피 및 인삼 저온숙성 복합 추출물의 제조방법 및 이를 포함하는 항노화 화장료 조성물 | |
CN109923122A (zh) | 新型肽以及包含其的用于抑制皮肤老化或皮肤皱纹形成的化妆料组合物 | |
CN102498128B (zh) | 具有转化生长因子的活性的肽及其用途 | |
CN114716515A (zh) | 一种多肽类似物及其制备方法和应用 | |
CN102558299B (zh) | 一种合成多肽及其应用 | |
CN109369781A (zh) | 一种麒麟菜抗氧化四肽及其应用 | |
CN108794574B (zh) | 一种抗氧化的活性肽及其应用 |
Legal Events
Date | Code | Title | Description |
---|---|---|---|
PB01 | Publication | ||
PB01 | Publication | ||
SE01 | Entry into force of request for substantive examination | ||
SE01 | Entry into force of request for substantive examination | ||
WW01 | Invention patent application withdrawn after publication |
Application publication date: 20180410 |
|
WW01 | Invention patent application withdrawn after publication |