JP6472580B2 - 皮膚細胞の増殖効果の増大した熱安定性のヒト上皮細胞成長因子−クモ毒の融合タンパク質及びこれを有効成分として含む皮膚しわの改善及び皮膚弾力の維持のための化粧料組成物 - Google Patents
皮膚細胞の増殖効果の増大した熱安定性のヒト上皮細胞成長因子−クモ毒の融合タンパク質及びこれを有効成分として含む皮膚しわの改善及び皮膚弾力の維持のための化粧料組成物 Download PDFInfo
- Publication number
- JP6472580B2 JP6472580B2 JP2018535797A JP2018535797A JP6472580B2 JP 6472580 B2 JP6472580 B2 JP 6472580B2 JP 2018535797 A JP2018535797 A JP 2018535797A JP 2018535797 A JP2018535797 A JP 2018535797A JP 6472580 B2 JP6472580 B2 JP 6472580B2
- Authority
- JP
- Japan
- Prior art keywords
- growth factor
- fusion protein
- epidermal growth
- human epidermal
- skin
- 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.)
- Active
Links
- 108020001507 fusion proteins Proteins 0.000 title claims description 105
- 239000002708 spider venom Substances 0.000 title claims description 105
- 102000037865 fusion proteins Human genes 0.000 title claims description 102
- 230000012010 growth Effects 0.000 title claims description 82
- 230000000694 effects Effects 0.000 title claims description 44
- 210000003491 skin Anatomy 0.000 title claims description 25
- 239000002537 cosmetic Substances 0.000 title claims description 20
- 239000004480 active ingredient Substances 0.000 title claims description 19
- 239000000203 mixture Substances 0.000 title claims description 19
- 230000004663 cell proliferation Effects 0.000 title claims description 17
- 230000037303 wrinkles Effects 0.000 title claims description 17
- 210000004927 skin cell Anatomy 0.000 title claims description 14
- 230000037394 skin elasticity Effects 0.000 title claims description 13
- 108090000623 proteins and genes Proteins 0.000 claims description 63
- 210000004027 cell Anatomy 0.000 claims description 57
- 241000588724 Escherichia coli Species 0.000 claims description 34
- 238000004519 manufacturing process Methods 0.000 claims description 30
- 239000013598 vector Substances 0.000 claims description 19
- 230000010261 cell growth Effects 0.000 claims description 13
- 230000035755 proliferation Effects 0.000 claims description 12
- FWMNVWWHGCHHJJ-SKKKGAJSSA-N 4-amino-1-[(2r)-6-amino-2-[[(2r)-2-[[(2r)-2-[[(2r)-2-amino-3-phenylpropanoyl]amino]-3-phenylpropanoyl]amino]-4-methylpentanoyl]amino]hexanoyl]piperidine-4-carboxylic acid Chemical compound C([C@H](C(=O)N[C@H](CC(C)C)C(=O)N[C@H](CCCCN)C(=O)N1CCC(N)(CC1)C(O)=O)NC(=O)[C@H](N)CC=1C=CC=CC=1)C1=CC=CC=C1 FWMNVWWHGCHHJJ-SKKKGAJSSA-N 0.000 claims description 11
- 108020004705 Codon Proteins 0.000 claims description 8
- 239000003102 growth factor Substances 0.000 claims description 6
- 230000001131 transforming effect Effects 0.000 claims description 3
- 241000239290 Araneae Species 0.000 claims description 2
- 210000001339 epidermal cell Anatomy 0.000 claims description 2
- 125000003275 alpha amino acid group Chemical group 0.000 claims 2
- 102000004169 proteins and genes Human genes 0.000 description 27
- 235000018102 proteins Nutrition 0.000 description 26
- 238000000034 method Methods 0.000 description 21
- 230000000052 comparative effect Effects 0.000 description 16
- 150000001413 amino acids Chemical group 0.000 description 13
- 238000012360 testing method Methods 0.000 description 12
- 230000014509 gene expression Effects 0.000 description 11
- 239000006071 cream Substances 0.000 description 10
- 239000006210 lotion Substances 0.000 description 10
- 102000009024 Epidermal Growth Factor Human genes 0.000 description 9
- 101500025419 Homo sapiens Epidermal growth factor Proteins 0.000 description 9
- 210000002950 fibroblast Anatomy 0.000 description 9
- 230000002500 effect on skin Effects 0.000 description 8
- 239000013604 expression vector Substances 0.000 description 8
- 229940116978 human epidermal growth factor Drugs 0.000 description 8
- GVUGOAYIVIDWIO-UFWWTJHBSA-N nepidermin Chemical compound C([C@@H](C(=O)N[C@@H]([C@@H](C)CC)C(=O)NCC(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CS)C(=O)N[C@@H](CCC(N)=O)C(=O)N[C@@H](CC=1C=CC(O)=CC=1)C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CC(O)=O)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CCCCN)C(=O)N[C@@H](CC=1C2=CC=CC=C2NC=1)C(=O)N[C@@H](CC=1C2=CC=CC=C2NC=1)C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CCCNC(N)=N)C(O)=O)NC(=O)CNC(=O)[C@@H](NC(=O)[C@@H](NC(=O)[C@H](CS)NC(=O)[C@H](CC(N)=O)NC(=O)[C@H](CS)NC(=O)[C@H](C)NC(=O)[C@H](CC=1C=CC(O)=CC=1)NC(=O)[C@H](CCCCN)NC(=O)[C@H](CC(O)=O)NC(=O)[C@H](CC(C)C)NC(=O)[C@H](C)NC(=O)[C@H](CCC(O)=O)NC(=O)[C@@H](NC(=O)[C@H](CC=1C=CC(O)=CC=1)NC(=O)[C@H](CCSC)NC(=O)[C@H](CS)NC(=O)[C@@H](NC(=O)CNC(=O)[C@H](CC(O)=O)NC(=O)[C@H](CC=1NC=NC=1)NC(=O)[C@H](CC(C)C)NC(=O)[C@H](CS)NC(=O)[C@H](CC=1C=CC(O)=CC=1)NC(=O)CNC(=O)[C@H](CC(O)=O)NC(=O)[C@H](CC=1NC=NC=1)NC(=O)[C@H](CO)NC(=O)[C@H](CC(C)C)NC(=O)[C@H]1N(CCC1)C(=O)[C@H](CS)NC(=O)[C@H](CCC(O)=O)NC(=O)[C@H](CO)NC(=O)[C@H](CC(O)=O)NC(=O)[C@H](CO)NC(=O)[C@@H](N)CC(N)=O)C(C)C)[C@@H](C)CC)C(C)C)C(C)C)C1=CC=C(O)C=C1 GVUGOAYIVIDWIO-UFWWTJHBSA-N 0.000 description 8
- 239000013612 plasmid Substances 0.000 description 8
- 230000001953 sensory effect Effects 0.000 description 8
- 239000000243 solution Substances 0.000 description 8
- 239000002552 dosage form Substances 0.000 description 7
- 210000002919 epithelial cell Anatomy 0.000 description 7
- 239000002773 nucleotide Substances 0.000 description 7
- 125000003729 nucleotide group Chemical group 0.000 description 7
- -1 softeners Substances 0.000 description 7
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 description 6
- DNIAPMSPPWPWGF-UHFFFAOYSA-N Propylene glycol Chemical compound CC(O)CO DNIAPMSPPWPWGF-UHFFFAOYSA-N 0.000 description 6
- 101710099833 Venom protein Proteins 0.000 description 6
- RAXXELZNTBOGNW-UHFFFAOYSA-N imidazole Natural products C1=CNC=N1 RAXXELZNTBOGNW-UHFFFAOYSA-N 0.000 description 6
- 230000006698 induction Effects 0.000 description 6
- 239000000686 essence Substances 0.000 description 5
- 244000005700 microbiome Species 0.000 description 5
- 239000000843 powder Substances 0.000 description 5
- 108090000765 processed proteins & peptides Proteins 0.000 description 5
- PXHVJJICTQNCMI-UHFFFAOYSA-N Nickel Chemical compound [Ni] PXHVJJICTQNCMI-UHFFFAOYSA-N 0.000 description 4
- 101000795829 Phoneutria nigriventer Delta-ctenitoxin-Pn2a Proteins 0.000 description 4
- VYPSYNLAJGMNEJ-UHFFFAOYSA-N Silicium dioxide Chemical compound O=[Si]=O VYPSYNLAJGMNEJ-UHFFFAOYSA-N 0.000 description 4
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 description 4
- 230000021615 conjugation Effects 0.000 description 4
- 239000012634 fragment Substances 0.000 description 4
- 230000006870 function Effects 0.000 description 4
- 238000003259 recombinant expression Methods 0.000 description 4
- 210000001626 skin fibroblast Anatomy 0.000 description 4
- 239000000725 suspension Substances 0.000 description 4
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 4
- 230000029663 wound healing Effects 0.000 description 4
- 102000001301 EGF receptor Human genes 0.000 description 3
- 108060006698 EGF receptor Proteins 0.000 description 3
- 101800003838 Epidermal growth factor Proteins 0.000 description 3
- 102400001368 Epidermal growth factor Human genes 0.000 description 3
- XEKOWRVHYACXOJ-UHFFFAOYSA-N Ethyl acetate Chemical compound CCOC(C)=O XEKOWRVHYACXOJ-UHFFFAOYSA-N 0.000 description 3
- GUBGYTABKSRVRQ-QKKXKWKRSA-N Lactose Natural products OC[C@H]1O[C@@H](O[C@H]2[C@H](O)[C@@H](O)C(O)O[C@@H]2CO)[C@H](O)[C@@H](O)[C@H]1O GUBGYTABKSRVRQ-QKKXKWKRSA-N 0.000 description 3
- 238000007792 addition Methods 0.000 description 3
- 235000001014 amino acid Nutrition 0.000 description 3
- 238000012258 culturing Methods 0.000 description 3
- 238000012217 deletion Methods 0.000 description 3
- 230000037430 deletion Effects 0.000 description 3
- 239000000839 emulsion Substances 0.000 description 3
- 229940116977 epidermal growth factor Drugs 0.000 description 3
- 210000000981 epithelium Anatomy 0.000 description 3
- 238000010438 heat treatment Methods 0.000 description 3
- 230000006872 improvement Effects 0.000 description 3
- 239000004615 ingredient Substances 0.000 description 3
- 239000008101 lactose Substances 0.000 description 3
- 238000012423 maintenance Methods 0.000 description 3
- 108020004707 nucleic acids Proteins 0.000 description 3
- 102000039446 nucleic acids Human genes 0.000 description 3
- 150000007523 nucleic acids Chemical class 0.000 description 3
- 235000016709 nutrition Nutrition 0.000 description 3
- 230000035764 nutrition Effects 0.000 description 3
- 229920002401 polyacrylamide Polymers 0.000 description 3
- 108091033319 polynucleotide Proteins 0.000 description 3
- 102000040430 polynucleotide Human genes 0.000 description 3
- 239000002157 polynucleotide Substances 0.000 description 3
- 229920001184 polypeptide Polymers 0.000 description 3
- 102000004196 processed proteins & peptides Human genes 0.000 description 3
- 108091008146 restriction endonucleases Proteins 0.000 description 3
- 239000002904 solvent Substances 0.000 description 3
- 239000007921 spray Substances 0.000 description 3
- VBEQCZHXXJYVRD-GACYYNSASA-N uroanthelone Chemical compound C([C@@H](C(=O)N[C@H](C(=O)N[C@@H](CS)C(=O)N[C@@H](CC(N)=O)C(=O)N[C@@H](CS)C(=O)N[C@H](C(=O)N[C@@H]([C@@H](C)CC)C(=O)NCC(=O)N[C@@H](CC=1C=CC(O)=CC=1)C(=O)N[C@@H](CO)C(=O)NCC(=O)N[C@@H](CC(O)=O)C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CS)C(=O)N[C@@H](CCC(N)=O)C(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CC(O)=O)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CC=1C2=CC=CC=C2NC=1)C(=O)N[C@@H](CC=1C2=CC=CC=C2NC=1)C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CCCNC(N)=N)C(O)=O)C(C)C)[C@@H](C)O)NC(=O)[C@H](CO)NC(=O)[C@H](CC(O)=O)NC(=O)[C@H](CC(C)C)NC(=O)[C@H](CO)NC(=O)[C@H](CCC(O)=O)NC(=O)[C@@H](NC(=O)[C@H](CC=1NC=NC=1)NC(=O)[C@H](CCSC)NC(=O)[C@H](CS)NC(=O)[C@@H](NC(=O)CNC(=O)CNC(=O)[C@H](CC(N)=O)NC(=O)[C@H](CC(C)C)NC(=O)[C@H](CS)NC(=O)[C@H](CC=1C=CC(O)=CC=1)NC(=O)CNC(=O)[C@H](CC(O)=O)NC(=O)[C@H](CC=1C=CC(O)=CC=1)NC(=O)[C@H](CO)NC(=O)[C@H](CO)NC(=O)[C@H]1N(CCC1)C(=O)[C@H](CS)NC(=O)CNC(=O)[C@H]1N(CCC1)C(=O)[C@H](CC=1C=CC(O)=CC=1)NC(=O)[C@H](CO)NC(=O)[C@@H](N)CC(N)=O)C(C)C)[C@@H](C)CC)C1=CC=C(O)C=C1 VBEQCZHXXJYVRD-GACYYNSASA-N 0.000 description 3
- 229920000936 Agarose Polymers 0.000 description 2
- 108091003079 Bovine Serum Albumin Proteins 0.000 description 2
- 108700010070 Codon Usage Proteins 0.000 description 2
- LCGLNKUTAGEVQW-UHFFFAOYSA-N Dimethyl ether Chemical compound COC LCGLNKUTAGEVQW-UHFFFAOYSA-N 0.000 description 2
- PEDCQBHIVMGVHV-UHFFFAOYSA-N Glycerol Natural products OCC(O)CO PEDCQBHIVMGVHV-UHFFFAOYSA-N 0.000 description 2
- KFZMGEQAYNKOFK-UHFFFAOYSA-N Isopropanol Chemical compound CC(C)O KFZMGEQAYNKOFK-UHFFFAOYSA-N 0.000 description 2
- QIVBCDIJIAJPQS-VIFPVBQESA-N L-tryptophane Chemical compound C1=CC=C2C(C[C@H](N)C(O)=O)=CNC2=C1 QIVBCDIJIAJPQS-VIFPVBQESA-N 0.000 description 2
- 239000002202 Polyethylene glycol Substances 0.000 description 2
- WCUXLLCKKVVCTQ-UHFFFAOYSA-M Potassium chloride Chemical compound [Cl-].[K+] WCUXLLCKKVVCTQ-UHFFFAOYSA-M 0.000 description 2
- ATUOYWHBWRKTHZ-UHFFFAOYSA-N Propane Chemical compound CCC ATUOYWHBWRKTHZ-UHFFFAOYSA-N 0.000 description 2
- 240000004808 Saccharomyces cerevisiae Species 0.000 description 2
- BNRNXUUZRGQAQC-UHFFFAOYSA-N Sildenafil Natural products CCCC1=NN(C)C(C(N2)=O)=C1N=C2C(C(=CC=1)OCC)=CC=1S(=O)(=O)N1CCN(C)CC1 BNRNXUUZRGQAQC-UHFFFAOYSA-N 0.000 description 2
- 206010040880 Skin irritation Diseases 0.000 description 2
- 108010052164 Sodium Channels Proteins 0.000 description 2
- 102000018674 Sodium Channels Human genes 0.000 description 2
- QIVBCDIJIAJPQS-UHFFFAOYSA-N Tryptophan Natural products C1=CC=C2C(CC(N)C(O)=O)=CNC2=C1 QIVBCDIJIAJPQS-UHFFFAOYSA-N 0.000 description 2
- XSQUKJJJFZCRTK-UHFFFAOYSA-N Urea Chemical compound NC(N)=O XSQUKJJJFZCRTK-UHFFFAOYSA-N 0.000 description 2
- 241000700605 Viruses Species 0.000 description 2
- XLOMVQKBTHCTTD-UHFFFAOYSA-N Zinc monoxide Chemical compound [Zn]=O XLOMVQKBTHCTTD-UHFFFAOYSA-N 0.000 description 2
- 230000009471 action Effects 0.000 description 2
- 239000000440 bentonite Substances 0.000 description 2
- 229910000278 bentonite Inorganic materials 0.000 description 2
- SVPXDRXYRYOSEX-UHFFFAOYSA-N bentoquatam Chemical compound O.O=[Si]=O.O=[Al]O[Al]=O SVPXDRXYRYOSEX-UHFFFAOYSA-N 0.000 description 2
- SESFRYSPDFLNCH-UHFFFAOYSA-N benzyl benzoate Chemical compound C=1C=CC=CC=1C(=O)OCC1=CC=CC=C1 SESFRYSPDFLNCH-UHFFFAOYSA-N 0.000 description 2
- 230000036760 body temperature Effects 0.000 description 2
- 229940098773 bovine serum albumin Drugs 0.000 description 2
- 229940041514 candida albicans extract Drugs 0.000 description 2
- 239000012094 cell viability reagent Substances 0.000 description 2
- 239000013078 crystal Substances 0.000 description 2
- 238000003235 crystal violet staining Methods 0.000 description 2
- 210000004207 dermis Anatomy 0.000 description 2
- 238000001514 detection method Methods 0.000 description 2
- 238000004043 dyeing Methods 0.000 description 2
- 238000001962 electrophoresis Methods 0.000 description 2
- 238000004520 electroporation Methods 0.000 description 2
- 238000010828 elution Methods 0.000 description 2
- 210000002615 epidermis Anatomy 0.000 description 2
- 210000003527 eukaryotic cell Anatomy 0.000 description 2
- 238000001914 filtration Methods 0.000 description 2
- 230000004927 fusion Effects 0.000 description 2
- 239000000499 gel Substances 0.000 description 2
- 235000013922 glutamic acid Nutrition 0.000 description 2
- 239000004220 glutamic acid Substances 0.000 description 2
- 238000002955 isolation Methods 0.000 description 2
- BPHPUYQFMNQIOC-NXRLNHOXSA-N isopropyl beta-D-thiogalactopyranoside Chemical compound CC(C)S[C@@H]1O[C@H](CO)[C@H](O)[C@H](O)[C@H]1O BPHPUYQFMNQIOC-NXRLNHOXSA-N 0.000 description 2
- 239000006166 lysate Substances 0.000 description 2
- 239000003550 marker Substances 0.000 description 2
- 238000005259 measurement Methods 0.000 description 2
- 239000002609 medium Substances 0.000 description 2
- 239000012528 membrane Substances 0.000 description 2
- 229910052759 nickel Inorganic materials 0.000 description 2
- 239000003921 oil Substances 0.000 description 2
- 235000019198 oils Nutrition 0.000 description 2
- 239000006072 paste Substances 0.000 description 2
- 230000002085 persistent effect Effects 0.000 description 2
- 239000002953 phosphate buffered saline Substances 0.000 description 2
- 229920001223 polyethylene glycol Polymers 0.000 description 2
- 238000003752 polymerase chain reaction Methods 0.000 description 2
- 230000008569 process Effects 0.000 description 2
- 238000012545 processing Methods 0.000 description 2
- 210000001236 prokaryotic cell Anatomy 0.000 description 2
- 230000004952 protein activity Effects 0.000 description 2
- DEIYFTQMQPDXOT-UHFFFAOYSA-N sildenafil citrate Chemical compound OC(=O)CC(O)(C(O)=O)CC(O)=O.CCCC1=NN(C)C(C(N2)=O)=C1N=C2C(C(=CC=1)OCC)=CC=1S(=O)(=O)N1CCN(C)CC1 DEIYFTQMQPDXOT-UHFFFAOYSA-N 0.000 description 2
- 239000000377 silicon dioxide Substances 0.000 description 2
- 230000009759 skin aging Effects 0.000 description 2
- 230000036556 skin irritation Effects 0.000 description 2
- 231100000475 skin irritation Toxicity 0.000 description 2
- 239000011780 sodium chloride Substances 0.000 description 2
- 238000010186 staining Methods 0.000 description 2
- 230000001954 sterilising effect Effects 0.000 description 2
- 238000004659 sterilization and disinfection Methods 0.000 description 2
- 239000000454 talc Substances 0.000 description 2
- 229910052623 talc Inorganic materials 0.000 description 2
- CFMYXEVWODSLAX-QOZOJKKESA-N tetrodotoxin Chemical compound O([C@@]([C@H]1O)(O)O[C@H]2[C@@]3(O)CO)[C@H]3[C@@H](O)[C@]11[C@H]2[C@@H](O)N=C(N)N1 CFMYXEVWODSLAX-QOZOJKKESA-N 0.000 description 2
- 229950010357 tetrodotoxin Drugs 0.000 description 2
- CFMYXEVWODSLAX-UHFFFAOYSA-N tetrodotoxin Natural products C12C(O)NC(=N)NC2(C2O)C(O)C3C(CO)(O)C1OC2(O)O3 CFMYXEVWODSLAX-UHFFFAOYSA-N 0.000 description 2
- 230000009466 transformation Effects 0.000 description 2
- 229940094720 viagra Drugs 0.000 description 2
- 239000001993 wax Substances 0.000 description 2
- 239000012138 yeast extract Substances 0.000 description 2
- JNYAEWCLZODPBN-JGWLITMVSA-N (2r,3r,4s)-2-[(1r)-1,2-dihydroxyethyl]oxolane-3,4-diol Chemical compound OC[C@@H](O)[C@H]1OC[C@H](O)[C@H]1O JNYAEWCLZODPBN-JGWLITMVSA-N 0.000 description 1
- WNWHHMBRJJOGFJ-UHFFFAOYSA-N 16-methylheptadecan-1-ol Chemical class CC(C)CCCCCCCCCCCCCCCO WNWHHMBRJJOGFJ-UHFFFAOYSA-N 0.000 description 1
- JKMHFZQWWAIEOD-UHFFFAOYSA-N 2-[4-(2-hydroxyethyl)piperazin-1-yl]ethanesulfonic acid Chemical compound OCC[NH+]1CCN(CCS([O-])(=O)=O)CC1 JKMHFZQWWAIEOD-UHFFFAOYSA-N 0.000 description 1
- GUBGYTABKSRVRQ-XLOQQCSPSA-N Alpha-Lactose Chemical compound O[C@@H]1[C@@H](O)[C@@H](O)[C@@H](CO)O[C@H]1O[C@@H]1[C@@H](CO)O[C@H](O)[C@H](O)[C@H]1O GUBGYTABKSRVRQ-XLOQQCSPSA-N 0.000 description 1
- 241000193830 Bacillus <bacterium> Species 0.000 description 1
- 244000063299 Bacillus subtilis Species 0.000 description 1
- 235000014469 Bacillus subtilis Nutrition 0.000 description 1
- 241000193388 Bacillus thuringiensis Species 0.000 description 1
- 241000894006 Bacteria Species 0.000 description 1
- 241000699802 Cricetulus griseus Species 0.000 description 1
- FBPFZTCFMRRESA-FSIIMWSLSA-N D-Glucitol Natural products OC[C@H](O)[C@H](O)[C@@H](O)[C@H](O)CO FBPFZTCFMRRESA-FSIIMWSLSA-N 0.000 description 1
- 230000006820 DNA synthesis Effects 0.000 description 1
- 229920002307 Dextran Polymers 0.000 description 1
- 241000305071 Enterobacterales Species 0.000 description 1
- 208000010228 Erectile Dysfunction Diseases 0.000 description 1
- 206010015150 Erythema Diseases 0.000 description 1
- 241001302584 Escherichia coli str. K-12 substr. W3110 Species 0.000 description 1
- 102000010834 Extracellular Matrix Proteins Human genes 0.000 description 1
- 108010037362 Extracellular Matrix Proteins Proteins 0.000 description 1
- WQZGKKKJIJFFOK-GASJEMHNSA-N Glucose Natural products OC[C@H]1OC(O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-GASJEMHNSA-N 0.000 description 1
- WHUUTDBJXJRKMK-UHFFFAOYSA-N Glutamic acid Natural products OC(=O)C(N)CCC(O)=O WHUUTDBJXJRKMK-UHFFFAOYSA-N 0.000 description 1
- 239000007995 HEPES buffer Substances 0.000 description 1
- 241000238631 Hexapoda Species 0.000 description 1
- WHUUTDBJXJRKMK-VKHMYHEASA-N L-glutamic acid Chemical compound OC(=O)[C@@H](N)CCC(O)=O WHUUTDBJXJRKMK-VKHMYHEASA-N 0.000 description 1
- 229920000168 Microcrystalline cellulose Polymers 0.000 description 1
- 108091028043 Nucleic acid sequence Proteins 0.000 description 1
- 108700026244 Open Reading Frames Proteins 0.000 description 1
- 208000002193 Pain Diseases 0.000 description 1
- 241000238907 Phoneutria nigriventer Species 0.000 description 1
- 229920003171 Poly (ethylene oxide) Polymers 0.000 description 1
- 239000004952 Polyamide Substances 0.000 description 1
- 206010036790 Productive cough Diseases 0.000 description 1
- 108010076504 Protein Sorting Signals Proteins 0.000 description 1
- 102000004022 Protein-Tyrosine Kinases Human genes 0.000 description 1
- 108090000412 Protein-Tyrosine Kinases Proteins 0.000 description 1
- 208000003251 Pruritus Diseases 0.000 description 1
- 241000589516 Pseudomonas Species 0.000 description 1
- 108020004511 Recombinant DNA Proteins 0.000 description 1
- 108020005091 Replication Origin Proteins 0.000 description 1
- 241000293869 Salmonella enterica subsp. enterica serovar Typhimurium Species 0.000 description 1
- 241000607715 Serratia marcescens Species 0.000 description 1
- XUIMIQQOPSSXEZ-UHFFFAOYSA-N Silicon Chemical compound [Si] XUIMIQQOPSSXEZ-UHFFFAOYSA-N 0.000 description 1
- FKNQFGJONOIPTF-UHFFFAOYSA-N Sodium cation Chemical compound [Na+] FKNQFGJONOIPTF-UHFFFAOYSA-N 0.000 description 1
- 229920002472 Starch Polymers 0.000 description 1
- 206010052428 Wound Diseases 0.000 description 1
- 208000027418 Wounds and injury Diseases 0.000 description 1
- OIPILFWXSMYKGL-UHFFFAOYSA-N acetylcholine Chemical compound CC(=O)OCC[N+](C)(C)C OIPILFWXSMYKGL-UHFFFAOYSA-N 0.000 description 1
- 229960004373 acetylcholine Drugs 0.000 description 1
- 239000013543 active substance Substances 0.000 description 1
- 239000002671 adjuvant Substances 0.000 description 1
- 230000032683 aging Effects 0.000 description 1
- XAGFODPZIPBFFR-UHFFFAOYSA-N aluminium Chemical compound [Al] XAGFODPZIPBFFR-UHFFFAOYSA-N 0.000 description 1
- 229910052782 aluminium Inorganic materials 0.000 description 1
- WNROFYMDJYEPJX-UHFFFAOYSA-K aluminium hydroxide Chemical compound [OH-].[OH-].[OH-].[Al+3] WNROFYMDJYEPJX-UHFFFAOYSA-K 0.000 description 1
- 238000004458 analytical method Methods 0.000 description 1
- 239000010775 animal oil Substances 0.000 description 1
- 230000000692 anti-sense effect Effects 0.000 description 1
- 239000003963 antioxidant agent Substances 0.000 description 1
- 229940097012 bacillus thuringiensis Drugs 0.000 description 1
- 230000001580 bacterial effect Effects 0.000 description 1
- 229960002903 benzyl benzoate Drugs 0.000 description 1
- 239000012148 binding buffer Substances 0.000 description 1
- 230000004071 biological effect Effects 0.000 description 1
- 230000008827 biological function Effects 0.000 description 1
- 230000015572 biosynthetic process Effects 0.000 description 1
- 239000002981 blocking agent Substances 0.000 description 1
- 239000000872 buffer Substances 0.000 description 1
- 239000008366 buffered solution Substances 0.000 description 1
- 239000001273 butane Substances 0.000 description 1
- 239000001506 calcium phosphate Substances 0.000 description 1
- 229910000389 calcium phosphate Inorganic materials 0.000 description 1
- 235000011010 calcium phosphates Nutrition 0.000 description 1
- 239000000378 calcium silicate Substances 0.000 description 1
- 229910052918 calcium silicate Inorganic materials 0.000 description 1
- OYACROKNLOSFPA-UHFFFAOYSA-N calcium;dioxido(oxo)silane Chemical compound [Ca+2].[O-][Si]([O-])=O OYACROKNLOSFPA-UHFFFAOYSA-N 0.000 description 1
- 239000004202 carbamide Substances 0.000 description 1
- 125000003178 carboxy group Chemical group [H]OC(*)=O 0.000 description 1
- 239000000969 carrier Substances 0.000 description 1
- 239000006143 cell culture medium Substances 0.000 description 1
- 239000013592 cell lysate Substances 0.000 description 1
- 230000012292 cell migration Effects 0.000 description 1
- 239000001913 cellulose Substances 0.000 description 1
- 229920002678 cellulose Polymers 0.000 description 1
- 235000010980 cellulose Nutrition 0.000 description 1
- 238000005119 centrifugation Methods 0.000 description 1
- 239000002738 chelating agent Substances 0.000 description 1
- 239000003795 chemical substances by application Substances 0.000 description 1
- 210000004978 chinese hamster ovary cell Anatomy 0.000 description 1
- 150000005827 chlorofluoro hydrocarbons Chemical class 0.000 description 1
- 239000011248 coating agent Substances 0.000 description 1
- 238000000576 coating method Methods 0.000 description 1
- 210000005080 cortical synaptosome Anatomy 0.000 description 1
- 210000004748 cultured cell Anatomy 0.000 description 1
- 230000007423 decrease Effects 0.000 description 1
- 238000010586 diagram Methods 0.000 description 1
- 235000014113 dietary fatty acids Nutrition 0.000 description 1
- 239000003085 diluting agent Substances 0.000 description 1
- 238000006471 dimerization reaction Methods 0.000 description 1
- LOKCTEFSRHRXRJ-UHFFFAOYSA-I dipotassium trisodium dihydrogen phosphate hydrogen phosphate dichloride Chemical compound P(=O)(O)(O)[O-].[K+].P(=O)(O)([O-])[O-].[Na+].[Na+].[Cl-].[K+].[Cl-].[Na+] LOKCTEFSRHRXRJ-UHFFFAOYSA-I 0.000 description 1
- BNIILDVGGAEEIG-UHFFFAOYSA-L disodium hydrogen phosphate Chemical compound [Na+].[Na+].OP([O-])([O-])=O BNIILDVGGAEEIG-UHFFFAOYSA-L 0.000 description 1
- 239000000975 dye Substances 0.000 description 1
- 239000003974 emollient agent Substances 0.000 description 1
- 239000003995 emulsifying agent Substances 0.000 description 1
- 230000008472 epithelial growth Effects 0.000 description 1
- 231100000321 erythema Toxicity 0.000 description 1
- 238000011156 evaluation Methods 0.000 description 1
- 238000002474 experimental method Methods 0.000 description 1
- 210000002744 extracellular matrix Anatomy 0.000 description 1
- 239000000194 fatty acid Substances 0.000 description 1
- 229930195729 fatty acid Natural products 0.000 description 1
- 239000000945 filler Substances 0.000 description 1
- 239000006260 foam Substances 0.000 description 1
- 239000004088 foaming agent Substances 0.000 description 1
- 239000003205 fragrance Substances 0.000 description 1
- 238000001502 gel electrophoresis Methods 0.000 description 1
- 239000003349 gelling agent Substances 0.000 description 1
- 230000002068 genetic effect Effects 0.000 description 1
- 239000008103 glucose Substances 0.000 description 1
- 150000002307 glutamic acids Chemical class 0.000 description 1
- 230000034659 glycolysis Effects 0.000 description 1
- 239000008269 hand cream Substances 0.000 description 1
- 210000005260 human cell Anatomy 0.000 description 1
- 201000001881 impotence Diseases 0.000 description 1
- 238000000338 in vitro Methods 0.000 description 1
- 238000001727 in vivo Methods 0.000 description 1
- 230000002779 inactivation Effects 0.000 description 1
- 210000003000 inclusion body Anatomy 0.000 description 1
- 208000015181 infectious disease Diseases 0.000 description 1
- 230000004941 influx Effects 0.000 description 1
- 210000004692 intercellular junction Anatomy 0.000 description 1
- 230000003834 intracellular effect Effects 0.000 description 1
- 230000008316 intracellular mechanism Effects 0.000 description 1
- 230000031146 intracellular signal transduction Effects 0.000 description 1
- 230000007803 itching Effects 0.000 description 1
- 101150109249 lacI gene Proteins 0.000 description 1
- 150000002632 lipids Chemical class 0.000 description 1
- 239000002502 liposome Substances 0.000 description 1
- 239000007788 liquid Substances 0.000 description 1
- 230000014759 maintenance of location Effects 0.000 description 1
- 210000004962 mammalian cell Anatomy 0.000 description 1
- 230000001404 mediated effect Effects 0.000 description 1
- 108020004999 messenger RNA Proteins 0.000 description 1
- 238000001471 micro-filtration Methods 0.000 description 1
- 235000019813 microcrystalline cellulose Nutrition 0.000 description 1
- 239000008108 microcrystalline cellulose Substances 0.000 description 1
- 229940016286 microcrystalline cellulose Drugs 0.000 description 1
- 238000000520 microinjection Methods 0.000 description 1
- 238000013508 migration Methods 0.000 description 1
- 235000013336 milk Nutrition 0.000 description 1
- 239000008267 milk Substances 0.000 description 1
- 210000004080 milk Anatomy 0.000 description 1
- CQDGTJPVBWZJAZ-UHFFFAOYSA-N monoethyl carbonate Chemical compound CCOC(O)=O CQDGTJPVBWZJAZ-UHFFFAOYSA-N 0.000 description 1
- 229910000402 monopotassium phosphate Inorganic materials 0.000 description 1
- 235000019796 monopotassium phosphate Nutrition 0.000 description 1
- IJDNQMDRQITEOD-UHFFFAOYSA-N n-butane Chemical compound CCCC IJDNQMDRQITEOD-UHFFFAOYSA-N 0.000 description 1
- OFBQJSOFQDEBGM-UHFFFAOYSA-N n-pentane Natural products CCCCC OFBQJSOFQDEBGM-UHFFFAOYSA-N 0.000 description 1
- 239000013642 negative control Substances 0.000 description 1
- 239000012053 oil suspension Substances 0.000 description 1
- 239000003960 organic solvent Substances 0.000 description 1
- 210000001672 ovary Anatomy 0.000 description 1
- 206010033675 panniculitis Diseases 0.000 description 1
- 239000012188 paraffin wax Substances 0.000 description 1
- 230000035515 penetration Effects 0.000 description 1
- WVDDGKGOMKODPV-ZQBYOMGUSA-N phenyl(114C)methanol Chemical compound O[14CH2]C1=CC=CC=C1 WVDDGKGOMKODPV-ZQBYOMGUSA-N 0.000 description 1
- PJNZPQUBCPKICU-UHFFFAOYSA-N phosphoric acid;potassium Chemical compound [K].OP(O)(O)=O PJNZPQUBCPKICU-UHFFFAOYSA-N 0.000 description 1
- 239000000049 pigment Substances 0.000 description 1
- 229920002647 polyamide Polymers 0.000 description 1
- 229920000136 polysorbate Polymers 0.000 description 1
- 239000001103 potassium chloride Substances 0.000 description 1
- 235000011164 potassium chloride Nutrition 0.000 description 1
- 239000008057 potassium phosphate buffer Substances 0.000 description 1
- 238000001556 precipitation Methods 0.000 description 1
- 239000003755 preservative agent Substances 0.000 description 1
- 125000002924 primary amino group Chemical group [H]N([H])* 0.000 description 1
- 230000002062 proliferating effect Effects 0.000 description 1
- 239000001294 propane Substances 0.000 description 1
- 239000003380 propellant Substances 0.000 description 1
- 238000001243 protein synthesis Methods 0.000 description 1
- 238000000746 purification Methods 0.000 description 1
- 239000002994 raw material Substances 0.000 description 1
- 108020003175 receptors Proteins 0.000 description 1
- 102000005962 receptors Human genes 0.000 description 1
- 230000006798 recombination Effects 0.000 description 1
- 238000005215 recombination Methods 0.000 description 1
- 230000009467 reduction Effects 0.000 description 1
- 230000001105 regulatory effect Effects 0.000 description 1
- 230000010076 replication Effects 0.000 description 1
- 230000004044 response Effects 0.000 description 1
- 230000028327 secretion Effects 0.000 description 1
- 239000003352 sequestering agent Substances 0.000 description 1
- 239000010703 silicon Substances 0.000 description 1
- 229910052710 silicon Inorganic materials 0.000 description 1
- 230000036560 skin regeneration Effects 0.000 description 1
- 239000000344 soap Substances 0.000 description 1
- 239000011734 sodium Substances 0.000 description 1
- 229910001415 sodium ion Inorganic materials 0.000 description 1
- 239000011537 solubilization buffer Substances 0.000 description 1
- 239000000600 sorbitol Substances 0.000 description 1
- 210000003802 sputum Anatomy 0.000 description 1
- 208000024794 sputum Diseases 0.000 description 1
- 239000003381 stabilizer Substances 0.000 description 1
- 239000008107 starch Substances 0.000 description 1
- 235000019698 starch Nutrition 0.000 description 1
- 239000012536 storage buffer Substances 0.000 description 1
- 210000004304 subcutaneous tissue Anatomy 0.000 description 1
- 239000000126 substance Substances 0.000 description 1
- 238000006467 substitution reaction Methods 0.000 description 1
- 239000004094 surface-active agent Substances 0.000 description 1
- 239000000375 suspending agent Substances 0.000 description 1
- 238000003786 synthesis reaction Methods 0.000 description 1
- 239000002562 thickening agent Substances 0.000 description 1
- 238000013518 transcription Methods 0.000 description 1
- 230000035897 transcription Effects 0.000 description 1
- 230000002103 transcriptional effect Effects 0.000 description 1
- 230000014616 translation Effects 0.000 description 1
- 230000014621 translational initiation Effects 0.000 description 1
- QORWJWZARLRLPR-UHFFFAOYSA-H tricalcium bis(phosphate) Chemical compound [Ca+2].[Ca+2].[Ca+2].[O-]P([O-])([O-])=O.[O-]P([O-])([O-])=O QORWJWZARLRLPR-UHFFFAOYSA-H 0.000 description 1
- 235000015112 vegetable and seed oil Nutrition 0.000 description 1
- 239000008158 vegetable oil Substances 0.000 description 1
- 239000002435 venom Substances 0.000 description 1
- 210000001048 venom Anatomy 0.000 description 1
- 231100000611 venom Toxicity 0.000 description 1
- 239000011782 vitamin Substances 0.000 description 1
- 235000013343 vitamin Nutrition 0.000 description 1
- 229940088594 vitamin Drugs 0.000 description 1
- 229930003231 vitamin Natural products 0.000 description 1
- 239000000341 volatile oil Substances 0.000 description 1
- 238000005406 washing Methods 0.000 description 1
- 239000000080 wetting agent Substances 0.000 description 1
- 239000011787 zinc oxide Substances 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
- C07K14/475—Growth factors; Growth regulators
- C07K14/485—Epidermal growth factor [EGF], i.e. urogastrone
-
- 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
- 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
- A61K8/92—Oils, fats or waxes; Derivatives thereof, e.g. hydrogenation products thereof
- A61K8/925—Oils, fats or waxes; Derivatives thereof, e.g. hydrogenation products thereof of animal origin
-
- 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
-
- 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
- A61Q19/08—Anti-ageing preparations
-
- 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
-
- 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
- C07K14/43504—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans from invertebrates
- C07K14/43513—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans from invertebrates from arachnidae
- C07K14/43518—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans from invertebrates from arachnidae from spiders
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N15/00—Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
- C12N15/09—Recombinant DNA-technology
- C12N15/11—DNA or RNA fragments; Modified forms thereof; Non-coding nucleic acids having a biological activity
- C12N15/62—DNA sequences coding for fusion proteins
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K2800/00—Properties of cosmetic compositions or active ingredients thereof or formulation aids used therein and process related aspects
- A61K2800/74—Biological properties of particular ingredients
- A61K2800/78—Enzyme modulators, e.g. Enzyme agonists
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K2319/00—Fusion polypeptide
Landscapes
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Genetics & Genomics (AREA)
- Organic Chemistry (AREA)
- General Health & Medical Sciences (AREA)
- Zoology (AREA)
- Molecular Biology (AREA)
- Engineering & Computer Science (AREA)
- Gastroenterology & Hepatology (AREA)
- Biochemistry (AREA)
- Biophysics (AREA)
- Biomedical Technology (AREA)
- Public Health (AREA)
- Veterinary Medicine (AREA)
- Animal Behavior & Ethology (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- Toxicology (AREA)
- Medicinal Chemistry (AREA)
- Biotechnology (AREA)
- Insects & Arthropods (AREA)
- Wood Science & Technology (AREA)
- Bioinformatics & Cheminformatics (AREA)
- General Engineering & Computer Science (AREA)
- Dermatology (AREA)
- Birds (AREA)
- Epidemiology (AREA)
- Physics & Mathematics (AREA)
- Plant Pathology (AREA)
- Microbiology (AREA)
- Tropical Medicine & Parasitology (AREA)
- Gerontology & Geriatric Medicine (AREA)
- Oil, Petroleum & Natural Gas (AREA)
- Peptides Or Proteins (AREA)
- Cosmetics (AREA)
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
- Preparation Of Compounds By Using Micro-Organisms (AREA)
Description
ヒト上皮細胞成長因子−クモ毒の融合タンパク質をコードする最適化した遺伝子、組換え発現ベクター及び形質転換組換え微生物は、次の方法で製作した。
実施例1で製造したE.coli Rosetta2(DE3)pLysSをそれぞれ1LのLB培地(10%トリプトファン、10%塩化ナトリウム及び5%酵母抽出物)あるいはBSB培地(1%トリプトファン、0.5%酵母抽出物、1%グルコース及び0.1%HEPES(pH7.0)、(株)ネクスジェンバイオテク社製)を使って回分培養する時にOD600=0.6〜0.8になるまで、または20Lの発酵器を使って連続培養する時にはOD600=15〜20になるまで培養した。次いで、細胞培養培地にそれぞれ最終1〜5mM IPTGまたは2%ラクトースを加えて組換え大腸菌の遺伝子発現を誘導した。遺伝子発現の誘導後に3〜4時間さらに培養して細胞を遠心分離方法を用いて回収した。この細胞を緩衝溶液(Phosphate buffered saline、8gの塩化ナトリウム、0.2gの塩化カリウム、1.44gのリン酸水素二ナトリウム(Na2HPO4)、0.24gのリン酸二水素カリウム(KH2PO4)/L、pH7.4)に完全に懸濁させた後、超音波破砕機を使って細胞を破壊して細胞内タンパク質を含む溶液を分離した。
実施例2で分離及び精製したヒト上皮細胞成長因子−クモ毒の融合タンパク質の存在が確認された試料を選んでヒト上皮細胞成長因子−クモ毒の融合タンパク質の活性を測定した。
実施例2で分離及び精製したヒト上皮細胞成長因子−クモ毒の融合タンパク質の存在が確認された試料を選んで熱安定性を実験した。熱安定性実験は皮膚線維芽細胞にガンマ滅菌(35kgrayの照射)または高圧滅菌(121℃で15分間処理)したヒト上皮細胞成長因子−クモ毒の融合タンパク質をそれぞれ0.02〜0.2ppm濃度で処理し、3日間37℃で培養した後、クリスタルバイオレット染色を通じてタンパク質処理による細胞増殖度を比べて分析した。
実施例2で分離及び精製したヒト上皮細胞成長因子−クモ毒の融合タンパク質の存在が確認された試料を選んでヒト上皮細胞成長因子−クモ毒の融合タンパク質の活性を測定した。融合タンパク質の活性の測定はHaCaT細胞を培養し、培養された細胞に融合タンパク質を0、0.02ppm、0.2ppm、2ppm及び20ppm濃度で処理して培養細胞の細胞増殖、傷治癒及び細胞接合を分析して評価した。
前記実施例2で分離及び精製したヒト上皮細胞成長因子−クモ毒の融合タンパク質を有効成分として、製造例1、2、3及び4と比較例1、2、3及び4の化粧料組成物を製造して官能試験を施した。
ヒト上皮細胞成長因子−クモ毒の融合タンパク質を有効成分として添加し、下記の表1に記載の成分及び含量で製造例1のスキンを製造した。
ヒト上皮細胞成長因子−クモ毒の融合タンパク質を有効成分として添加し、下記の表3に記載の成分及び含量で製造例2のエッセンスを製造した。
ヒト上皮細胞成長因子−クモ毒の融合タンパク質を有効成分として添加し、下記の表5に記載の成分及び含量で製造例3のローションを製造した。また、ヒト上皮細胞成長因子−クモ毒の融合タンパク質を有効成分として添加せずに、下記の表5に記載の成分及び含量で比較例3のローションを製造した。
ヒト上皮細胞成長因子−クモ毒の融合タンパク質を有効成分として添加し、下記の表7に記載の成分及び含量で製造例4のクリームを製造した。また、ヒト上皮細胞成長因子−クモ毒の融合タンパク質を有効成分として添加せずに、下記の表7に記載の成分及び含量で比較例4のクリームを製造した。
Claims (8)
- 配列番号2のアミノ酸配列からなる皮膚細胞の増殖効果の増大した熱安定性のヒト上皮細胞成長因子−クモ毒の融合タンパク質。
- 請求項1に記載の皮膚細胞の増殖効果の増大した熱安定性のヒト上皮細胞成長因子−クモ毒の融合タンパク質をコードする遺伝子。
- 前記遺伝子は大腸菌コドンに最適化した配列番号1の塩基配列からなることを特徴とする、請求項2に記載の遺伝子。
- 請求項2または3に記載の遺伝子を含む組換えベクター。
- 請求項4に記載の組換えベクターに形質転換された宿主細胞。
- 請求項4に記載の組換えベクターで宿主細胞を形質転換させてヒト上皮細胞成長因子−クモ毒の融合タンパク質をコードする遺伝子を過発現させるステップを含む、宿主細胞でのヒト上皮細胞成長因子−クモ毒の融合タンパク質の生産方法。
- 前記宿主細胞は大腸菌であることを特徴とする、請求項6に記載のヒト上皮細胞成長因子−クモ毒の融合タンパク質の生産方法。
- 配列番号2のアミノ酸配列からなる皮膚細胞の増殖効果の増大した熱安定性のヒト上皮細胞成長因子−クモ毒の融合タンパク質を有効成分として含む、皮膚しわの改善及び皮膚弾力の維持のための化粧料組成物。
Applications Claiming Priority (3)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
KR1020160069272A KR101636851B1 (ko) | 2016-06-03 | 2016-06-03 | 피부 세포 증식 효과가 증가한 열 안정성 인간 상피세포성장인자-거미독 융합단백질 및 이를 유효성분으로 함유하는 피부 주름 개선 및 탄력 유지용 화장료 조성물 |
KR10-2016-0069272 | 2016-06-03 | ||
PCT/KR2016/007984 WO2017209347A1 (ko) | 2016-06-03 | 2016-07-22 | 피부 세포 증식 효과가 증가한 열 안정성 인간 상피세포성장인자-거미독 융합단백질 및 이를 유효성분으로 함유하는 피부 주름 개선 및 탄력 유지용 화장료 조성물 |
Publications (2)
Publication Number | Publication Date |
---|---|
JP2018529385A JP2018529385A (ja) | 2018-10-11 |
JP6472580B2 true JP6472580B2 (ja) | 2019-02-20 |
Family
ID=56502662
Family Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
JP2018535797A Active JP6472580B2 (ja) | 2016-06-03 | 2016-07-22 | 皮膚細胞の増殖効果の増大した熱安定性のヒト上皮細胞成長因子−クモ毒の融合タンパク質及びこれを有効成分として含む皮膚しわの改善及び皮膚弾力の維持のための化粧料組成物 |
Country Status (7)
Country | Link |
---|---|
US (1) | US10125181B2 (ja) |
EP (1) | EP3339319B1 (ja) |
JP (1) | JP6472580B2 (ja) |
KR (1) | KR101636851B1 (ja) |
CN (1) | CN108137663A (ja) |
ES (1) | ES2765282T3 (ja) |
WO (1) | WO2017209347A1 (ja) |
Families Citing this family (5)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
KR101613302B1 (ko) * | 2015-12-30 | 2016-04-18 | (주)넥스젠바이오텍 | Sv82 폴리펩타이드 및 이를 유효성분으로 함유하는 피부 주름 개선 및 탄력 유지용 화장료 조성물 |
KR101678393B1 (ko) * | 2016-08-25 | 2016-12-06 | (주)넥스젠바이오텍 | 피부 세포 증식 효과가 증가한 전갈독 융합단백질 및 이를 유효성분으로 함유하는 피부 주름 개선, 피부 탄력 증대 및 항노화 화장료 조성물 |
KR101678392B1 (ko) * | 2016-09-30 | 2016-11-22 | (주)넥스젠바이오텍 | 항산화 기능 및 피부 세포 증식 효과가 증가된 부동화 단백질 및 인간 상피세포성장인자를 포함하는 융합단백질 및 이를 유효성분으로 함유하는 피부 주름 개선용 화장료 조성물 |
US10905738B2 (en) * | 2018-07-05 | 2021-02-02 | Biozeus Desenvolvimento De Produtos Biofarmacêuticos | Synthetic peptides, prodrugs, pharmaceutical compositions and uses |
KR102101952B1 (ko) * | 2018-11-23 | 2020-04-27 | 주식회사 코씨드바이오팜 | 거미줄 추출물을 함유하는 피부 미백용 및 자외선에 의한 피부 손상 예방용 화장료 조성물 |
Family Cites Families (8)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US20070260039A1 (en) * | 2002-01-11 | 2007-11-08 | Karatzas Costas N | Methods of Producing Silk Polypeptides and Products Thereof |
KR20050027838A (ko) * | 2003-09-16 | 2005-03-21 | (주)넥스젠 | 식물체에서 발현된 재조합 인간 성장호르몬 |
KR100583350B1 (ko) * | 2004-11-03 | 2006-06-05 | (주)넥스젠 | Fas-1 도메인을 함유하는 융합 단백질을 이용한상피세포재생인자의 생산 방법 |
CN102757974B (zh) * | 2012-06-05 | 2014-07-23 | 陕西省微生物研究所 | 一种重组人表皮生长因子的新型制备方法 |
JP6430938B2 (ja) * | 2012-08-20 | 2018-11-28 | ウニベルシダーヂ フェデラル ヂ ミナス ジェライス−ウエフィエミジェー | 合成ポリペプチドPnTx(19)、医薬組成物及びその使用 |
KR101602689B1 (ko) | 2013-11-14 | 2016-03-11 | 주식회사 엘지생활건강 | 혈소판유래성장인자 b서브유닛의 융합단백질을 포함하는 피부개선용 화장료 조성물 |
CN103834664B (zh) * | 2014-03-27 | 2016-05-04 | 吉林大学 | 重组表皮生长因子egf及其制备方法 |
KR101613302B1 (ko) * | 2015-12-30 | 2016-04-18 | (주)넥스젠바이오텍 | Sv82 폴리펩타이드 및 이를 유효성분으로 함유하는 피부 주름 개선 및 탄력 유지용 화장료 조성물 |
-
2016
- 2016-06-03 KR KR1020160069272A patent/KR101636851B1/ko active IP Right Grant
- 2016-07-22 ES ES16904146T patent/ES2765282T3/es active Active
- 2016-07-22 CN CN201680060212.6A patent/CN108137663A/zh not_active Withdrawn
- 2016-07-22 US US15/522,159 patent/US10125181B2/en active Active
- 2016-07-22 WO PCT/KR2016/007984 patent/WO2017209347A1/ko active Application Filing
- 2016-07-22 JP JP2018535797A patent/JP6472580B2/ja active Active
- 2016-07-22 EP EP16904146.4A patent/EP3339319B1/en active Active
Also Published As
Publication number | Publication date |
---|---|
EP3339319B1 (en) | 2019-10-16 |
US20180186848A1 (en) | 2018-07-05 |
JP2018529385A (ja) | 2018-10-11 |
EP3339319A4 (en) | 2019-04-10 |
EP3339319A1 (en) | 2018-06-27 |
KR101636851B1 (ko) | 2016-07-06 |
US10125181B2 (en) | 2018-11-13 |
ES2765282T3 (es) | 2020-06-08 |
CN108137663A (zh) | 2018-06-08 |
WO2017209347A1 (ko) | 2017-12-07 |
Similar Documents
Publication | Publication Date | Title |
---|---|---|
JP6472580B2 (ja) | 皮膚細胞の増殖効果の増大した熱安定性のヒト上皮細胞成長因子−クモ毒の融合タンパク質及びこれを有効成分として含む皮膚しわの改善及び皮膚弾力の維持のための化粧料組成物 | |
JP6592112B2 (ja) | 皮膚細胞の増殖及び抗酸化効果の増大したボツリヌス毒素−ヒト上皮細胞成長因子の融合タンパク質及びこれを有効成分として含む皮膚再生及びしわ改善のための化粧料組成物 | |
JP6806889B2 (ja) | 抗酸化機能及び皮膚細胞増殖効果が増大した不凍タンパク質及びヒト上皮細胞成長因子を含む融合タンパク質並びにそれを有効成分として含有するしわ改善用化粧料組成物 | |
KR101652956B1 (ko) | 항산화 기능 및 세포 증식 효과가 증가된 인간 상피세포재생인자 융합단백질 및 이를 유효성분으로 함유하는 피부 주름 개선 및 항노화 화장료 조성물 | |
JP6261673B2 (ja) | 皮膚しわの改善及び弾力維持のための化粧料組成物 | |
CN112876569A (zh) | 一种rhTSG6-FNⅢ1-C融合蛋白及其在皮肤护理组合物中的用途和其制备方法 | |
US11166898B2 (en) | Human growth hormone fusion protein with enhanced thermal stability and cosmetic composition for anti-wrinkle and maintaining skin elasticity comprising the same as effective component | |
JP6770175B2 (ja) | 皮膚細胞の増殖効果が増大したサソリ毒融合タンパク質及びそれを有効成分として含有するシワ改善、皮膚弾力の増大及びアンチエイジング化粧料組成物 | |
KR101657298B1 (ko) | 세포 증식 효과가 증가한 인간 상피세포재생인자 융합단백질 및 이를 유효성분으로 함유하는 피부 주름 및 탄력 개선용 화장료 조성물 | |
KR101657299B1 (ko) | 세포 증식 효과가 증가한 인간 상피세포재생인자와 인간 성장호르몬의 융합단백질 및 이를 유효성분으로 함유하는 피부 주름 개선 및 항노화 화장료 조성물 | |
KR101784288B1 (ko) | 피부 세포 증식 효과가 증가한 성장 분화 인자 11과 열 충격 단백질 10의 융합단백질 및 이를 유효성분으로 함유하는 피부 주름 개선용 화장료 조성물 | |
KR102581377B1 (ko) | 콜라겐 합성 촉진 활성을 갖는 콜라겐 타입 4 저분자 펩타이드 및 이를 이용한 방법 | |
KR101776013B1 (ko) | 항산화 활성 및 피부 세포 증식 효과가 증가한 열 충격 단백질 10과 브라제인 단백질의 융합단백질 및 이를 유효성분으로 함유하는 피부 주름 개선용 화장료 조성물 | |
KR101800200B1 (ko) | 항산화 활성 및 피부 세포 증식 효과가 증가한 성장 분화 인자 11과 상피세포 성장인자의 융합단백질 및 이를 유효성분으로 함유하는 피부 주름 개선용 화장료 조성물 | |
KR101657297B1 (ko) | 세포 증식 효과가 증가한 인간 성장호르몬, 인간 상피세포재생인자 및 인간 인슐린유사성장인자-1을 포함하는 삼중 융합단백질 및 이를 유효성분으로 함유하는 피부 주름 개선 및 항노화 화장료 조성물 |
Legal Events
Date | Code | Title | Description |
---|---|---|---|
A621 | Written request for application examination |
Free format text: JAPANESE INTERMEDIATE CODE: A621 Effective date: 20180327 |
|
TRDD | Decision of grant or rejection written | ||
A01 | Written decision to grant a patent or to grant a registration (utility model) |
Free format text: JAPANESE INTERMEDIATE CODE: A01 Effective date: 20190108 |
|
A61 | First payment of annual fees (during grant procedure) |
Free format text: JAPANESE INTERMEDIATE CODE: A61 Effective date: 20190122 |
|
R150 | Certificate of patent or registration of utility model |
Ref document number: 6472580 Country of ref document: JP Free format text: JAPANESE INTERMEDIATE CODE: R150 |
|
R250 | Receipt of annual fees |
Free format text: JAPANESE INTERMEDIATE CODE: R250 |
|
R250 | Receipt of annual fees |
Free format text: JAPANESE INTERMEDIATE CODE: R250 |
|
R250 | Receipt of annual fees |
Free format text: JAPANESE INTERMEDIATE CODE: R250 |