KR20220074281A - The Peptide having Excellent Antioxidant Activity and Collagenase Inhibition Activity and Composition Containing the same - Google Patents
The Peptide having Excellent Antioxidant Activity and Collagenase Inhibition Activity and Composition Containing the same Download PDFInfo
- Publication number
- KR20220074281A KR20220074281A KR1020200162593A KR20200162593A KR20220074281A KR 20220074281 A KR20220074281 A KR 20220074281A KR 1020200162593 A KR1020200162593 A KR 1020200162593A KR 20200162593 A KR20200162593 A KR 20200162593A KR 20220074281 A KR20220074281 A KR 20220074281A
- Authority
- KR
- South Korea
- Prior art keywords
- group
- peptide
- resin
- trt
- activity
- Prior art date
Links
- 108090000765 processed proteins & peptides Proteins 0.000 title claims abstract description 55
- 102000029816 Collagenase Human genes 0.000 title claims abstract description 24
- 108060005980 Collagenase Proteins 0.000 title claims abstract description 24
- 229960002424 collagenase Drugs 0.000 title claims abstract description 24
- 239000000203 mixture Substances 0.000 title claims abstract description 23
- 230000003078 antioxidant effect Effects 0.000 title claims abstract description 21
- 230000000694 effects Effects 0.000 title description 11
- 230000005764 inhibitory process Effects 0.000 title description 4
- 230000002401 inhibitory effect Effects 0.000 claims abstract description 16
- 239000002537 cosmetic Substances 0.000 claims description 12
- 230000009759 skin aging Effects 0.000 claims description 9
- 230000037303 wrinkles Effects 0.000 claims description 9
- 235000013376 functional food Nutrition 0.000 claims description 6
- 230000036541 health Effects 0.000 claims description 6
- 230000001153 anti-wrinkle effect Effects 0.000 abstract description 5
- 230000003712 anti-aging effect Effects 0.000 abstract description 2
- -1 acetaminomethyl group Chemical group 0.000 description 58
- WYURNTSHIVDZCO-UHFFFAOYSA-N Tetrahydrofuran Chemical compound C1CCOC1 WYURNTSHIVDZCO-UHFFFAOYSA-N 0.000 description 48
- 239000011347 resin Substances 0.000 description 36
- 229920005989 resin Polymers 0.000 description 36
- 239000000243 solution Substances 0.000 description 28
- DTQVDTLACAAQTR-UHFFFAOYSA-N trifluoroacetic acid Substances OC(=O)C(F)(F)F DTQVDTLACAAQTR-UHFFFAOYSA-N 0.000 description 18
- NQRYJNQNLNOLGT-UHFFFAOYSA-N Piperidine Chemical compound C1CCNCC1 NQRYJNQNLNOLGT-UHFFFAOYSA-N 0.000 description 16
- 238000006243 chemical reaction Methods 0.000 description 15
- 238000002360 preparation method Methods 0.000 description 13
- 125000002221 trityl group Chemical group [H]C1=C([H])C([H])=C([H])C([H])=C1C([*])(C1=C(C(=C(C(=C1[H])[H])[H])[H])[H])C1=C([H])C([H])=C([H])C([H])=C1[H] 0.000 description 12
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 12
- 239000003875 Wang resin Substances 0.000 description 11
- YMWUJEATGCHHMB-UHFFFAOYSA-N Dichloromethane Chemical compound ClCCl YMWUJEATGCHHMB-UHFFFAOYSA-N 0.000 description 10
- ISWSIDIOOBJBQZ-UHFFFAOYSA-N Phenol Chemical compound OC1=CC=CC=C1 ISWSIDIOOBJBQZ-UHFFFAOYSA-N 0.000 description 10
- 150000001413 amino acids Chemical class 0.000 description 10
- 238000000034 method Methods 0.000 description 10
- 238000005406 washing Methods 0.000 description 10
- WEVYAHXRMPXWCK-UHFFFAOYSA-N Acetonitrile Chemical compound CC#N WEVYAHXRMPXWCK-UHFFFAOYSA-N 0.000 description 9
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 description 9
- OKKJLVBELUTLKV-UHFFFAOYSA-N Methanol Chemical compound OC OKKJLVBELUTLKV-UHFFFAOYSA-N 0.000 description 9
- 235000001014 amino acid Nutrition 0.000 description 9
- 239000006071 cream Substances 0.000 description 9
- 125000006239 protecting group Chemical group 0.000 description 9
- 125000001981 tert-butyldimethylsilyl group Chemical group [H]C([H])([H])[Si]([H])(C([H])([H])[H])[*]C(C([H])([H])[H])(C([H])([H])[H])C([H])([H])[H] 0.000 description 9
- QTBSBXVTEAMEQO-UHFFFAOYSA-N Acetic acid Chemical compound CC(O)=O QTBSBXVTEAMEQO-UHFFFAOYSA-N 0.000 description 8
- 108010035532 Collagen Proteins 0.000 description 8
- 102000008186 Collagen Human genes 0.000 description 8
- 229920001436 collagen Polymers 0.000 description 8
- 238000003756 stirring Methods 0.000 description 8
- KPFBUSLHFFWMAI-HYRPPVSQSA-N [(8r,9s,10r,13s,14s,17r)-17-acetyl-6-formyl-3-methoxy-10,13-dimethyl-1,2,7,8,9,11,12,14,15,16-decahydrocyclopenta[a]phenanthren-17-yl] acetate Chemical compound C1C[C@@H]2[C@](CCC(OC)=C3)(C)C3=C(C=O)C[C@H]2[C@@H]2CC[C@](OC(C)=O)(C(C)=O)[C@]21C KPFBUSLHFFWMAI-HYRPPVSQSA-N 0.000 description 7
- LCFXLZAXGXOXAP-QPJJXVBHSA-N ethyl (2e)-2-cyano-2-hydroxyiminoacetate Chemical compound CCOC(=O)C(=N\O)\C#N LCFXLZAXGXOXAP-QPJJXVBHSA-N 0.000 description 7
- 238000001914 filtration Methods 0.000 description 7
- 239000000463 material Substances 0.000 description 7
- 125000005931 tert-butyloxycarbonyl group Chemical group [H]C([H])([H])C(OC(*)=O)(C([H])([H])[H])C([H])([H])[H] 0.000 description 7
- 125000000025 triisopropylsilyl group Chemical group C(C)(C)[Si](C(C)C)(C(C)C)* 0.000 description 7
- 125000003903 2-propenyl group Chemical group [H]C([*])([H])C([H])=C([H])[H] 0.000 description 6
- LYCAIKOWRPUZTN-UHFFFAOYSA-N Ethylene glycol Chemical compound OCCO LYCAIKOWRPUZTN-UHFFFAOYSA-N 0.000 description 6
- KFZMGEQAYNKOFK-UHFFFAOYSA-N Isopropanol Chemical compound CC(C)O KFZMGEQAYNKOFK-UHFFFAOYSA-N 0.000 description 6
- 239000003963 antioxidant agent Substances 0.000 description 6
- 239000006210 lotion Substances 0.000 description 6
- IAZDPXIOMUYVGZ-UHFFFAOYSA-N Dimethylsulphoxide Chemical compound CS(C)=O IAZDPXIOMUYVGZ-UHFFFAOYSA-N 0.000 description 5
- 238000002965 ELISA Methods 0.000 description 5
- DHMQDGOQFOQNFH-UHFFFAOYSA-N Glycine Chemical compound NCC(O)=O DHMQDGOQFOQNFH-UHFFFAOYSA-N 0.000 description 5
- KDXKERNSBIXSRK-YFKPBYRVSA-N L-lysine Chemical compound NCCCC[C@H](N)C(O)=O KDXKERNSBIXSRK-YFKPBYRVSA-N 0.000 description 5
- 238000002835 absorbance Methods 0.000 description 5
- 210000003491 skin Anatomy 0.000 description 5
- 239000000126 substance Substances 0.000 description 5
- GQHTUMJGOHRCHB-UHFFFAOYSA-N 2,3,4,6,7,8,9,10-octahydropyrimido[1,2-a]azepine Chemical compound C1CCCCN2CCCN=C21 GQHTUMJGOHRCHB-UHFFFAOYSA-N 0.000 description 4
- HZAXFHJVJLSVMW-UHFFFAOYSA-N 2-Aminoethan-1-ol Chemical compound NCCO HZAXFHJVJLSVMW-UHFFFAOYSA-N 0.000 description 4
- HEDRZPFGACZZDS-UHFFFAOYSA-N Chloroform Chemical compound ClC(Cl)Cl HEDRZPFGACZZDS-UHFFFAOYSA-N 0.000 description 4
- BAVYZALUXZFZLV-UHFFFAOYSA-N Methylamine Chemical compound NC BAVYZALUXZFZLV-UHFFFAOYSA-N 0.000 description 4
- 108010050808 Procollagen Proteins 0.000 description 4
- OHDRQQURAXLVGJ-HLVWOLMTSA-N azane;(2e)-3-ethyl-2-[(e)-(3-ethyl-6-sulfo-1,3-benzothiazol-2-ylidene)hydrazinylidene]-1,3-benzothiazole-6-sulfonic acid Chemical compound [NH4+].[NH4+].S/1C2=CC(S([O-])(=O)=O)=CC=C2N(CC)C\1=N/N=C1/SC2=CC(S([O-])(=O)=O)=CC=C2N1CC OHDRQQURAXLVGJ-HLVWOLMTSA-N 0.000 description 4
- PAFZNILMFXTMIY-UHFFFAOYSA-N cyclohexylamine Chemical compound NC1CCCCC1 PAFZNILMFXTMIY-UHFFFAOYSA-N 0.000 description 4
- 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 4
- 235000013373 food additive Nutrition 0.000 description 4
- 239000002778 food additive Substances 0.000 description 4
- 238000004128 high performance liquid chromatography Methods 0.000 description 4
- 239000007788 liquid Substances 0.000 description 4
- 238000004519 manufacturing process Methods 0.000 description 4
- 239000002953 phosphate buffered saline Substances 0.000 description 4
- 102000004196 processed proteins & peptides Human genes 0.000 description 4
- 239000000047 product Substances 0.000 description 4
- 239000002904 solvent Substances 0.000 description 4
- 125000000999 tert-butyl group Chemical group [H]C([H])([H])C(*)(C([H])([H])[H])C([H])([H])[H] 0.000 description 4
- 150000003573 thiols Chemical group 0.000 description 4
- KLBPUVPNPAJWHZ-UMSFTDKQSA-N (2r)-2-(9h-fluoren-9-ylmethoxycarbonylamino)-3-tritylsulfanylpropanoic acid Chemical compound C([C@@H](C(=O)O)NC(=O)OCC1C2=CC=CC=C2C2=CC=CC=C21)SC(C=1C=CC=CC=1)(C=1C=CC=CC=1)C1=CC=CC=C1 KLBPUVPNPAJWHZ-UMSFTDKQSA-N 0.000 description 3
- PAQZWJGSJMLPMG-UHFFFAOYSA-N 2,4,6-tripropyl-1,3,5,2$l^{5},4$l^{5},6$l^{5}-trioxatriphosphinane 2,4,6-trioxide Chemical compound CCCP1(=O)OP(=O)(CCC)OP(=O)(CCC)O1 PAQZWJGSJMLPMG-UHFFFAOYSA-N 0.000 description 3
- RTZKZFJDLAIYFH-UHFFFAOYSA-N Diethyl ether Chemical compound CCOCC RTZKZFJDLAIYFH-UHFFFAOYSA-N 0.000 description 3
- WMFOQBRAJBCJND-UHFFFAOYSA-M Lithium hydroxide Chemical compound [Li+].[OH-] WMFOQBRAJBCJND-UHFFFAOYSA-M 0.000 description 3
- KDXKERNSBIXSRK-UHFFFAOYSA-N Lysine Natural products NCCCCC(N)C(O)=O KDXKERNSBIXSRK-UHFFFAOYSA-N 0.000 description 3
- KWYUFKZDYYNOTN-UHFFFAOYSA-M Potassium hydroxide Chemical compound [OH-].[K+] KWYUFKZDYYNOTN-UHFFFAOYSA-M 0.000 description 3
- RWRDLPDLKQPQOW-UHFFFAOYSA-N Pyrrolidine Chemical compound C1CCNC1 RWRDLPDLKQPQOW-UHFFFAOYSA-N 0.000 description 3
- HEMHJVSKTPXQMS-UHFFFAOYSA-M Sodium hydroxide Chemical compound [OH-].[Na+] HEMHJVSKTPXQMS-UHFFFAOYSA-M 0.000 description 3
- 125000006242 amine protecting group Chemical group 0.000 description 3
- PFKFTWBEEFSNDU-UHFFFAOYSA-N carbonyldiimidazole Chemical compound C1=CN=CN1C(=O)N1C=CN=C1 PFKFTWBEEFSNDU-UHFFFAOYSA-N 0.000 description 3
- 210000004207 dermis Anatomy 0.000 description 3
- 239000003085 diluting agent Substances 0.000 description 3
- 239000000686 essence Substances 0.000 description 3
- 239000001963 growth medium Substances 0.000 description 3
- 230000006872 improvement Effects 0.000 description 3
- 125000004184 methoxymethyl group Chemical group [H]C([H])([H])OC([H])([H])* 0.000 description 3
- GUVKYQNSMXSMMU-UHFFFAOYSA-N oxane Chemical group C1CCOCC1.C1CCOCC1 GUVKYQNSMXSMMU-UHFFFAOYSA-N 0.000 description 3
- 239000000546 pharmaceutical excipient Substances 0.000 description 3
- 239000000049 pigment Substances 0.000 description 3
- 239000000843 powder Substances 0.000 description 3
- 230000002265 prevention Effects 0.000 description 3
- 235000018102 proteins Nutrition 0.000 description 3
- 102000004169 proteins and genes Human genes 0.000 description 3
- 108090000623 proteins and genes Proteins 0.000 description 3
- 239000002994 raw material Substances 0.000 description 3
- 239000007787 solid Substances 0.000 description 3
- 238000003786 synthesis reaction Methods 0.000 description 3
- WHRNULOCNSKMGB-UHFFFAOYSA-N tetrahydrofuran thf Chemical group C1CCOC1.C1CCOC1 WHRNULOCNSKMGB-UHFFFAOYSA-N 0.000 description 3
- HNKJADCVZUBCPG-UHFFFAOYSA-N thioanisole Chemical compound CSC1=CC=CC=C1 HNKJADCVZUBCPG-UHFFFAOYSA-N 0.000 description 3
- XXMYDXUIZKNHDT-QNGWXLTQSA-N (2s)-2-(9h-fluoren-9-ylmethoxycarbonylamino)-3-(1-tritylimidazol-4-yl)propanoic acid Chemical compound C([C@@H](C(=O)O)NC(=O)OCC1C2=CC=CC=C2C2=CC=CC=C21)C(N=C1)=CN1C(C=1C=CC=CC=1)(C=1C=CC=CC=1)C1=CC=CC=C1 XXMYDXUIZKNHDT-QNGWXLTQSA-N 0.000 description 2
- IQQRAVYLUAZUGX-UHFFFAOYSA-N 1-butyl-3-methylimidazolium Chemical compound CCCCN1C=C[N+](C)=C1 IQQRAVYLUAZUGX-UHFFFAOYSA-N 0.000 description 2
- 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 2
- QOSSAOTZNIDXMA-UHFFFAOYSA-N Dicylcohexylcarbodiimide Chemical compound C1CCCCC1N=C=NC1CCCCC1 QOSSAOTZNIDXMA-UHFFFAOYSA-N 0.000 description 2
- 239000004471 Glycine Substances 0.000 description 2
- UFHFLCQGNIYNRP-UHFFFAOYSA-N Hydrogen Chemical compound [H][H] UFHFLCQGNIYNRP-UHFFFAOYSA-N 0.000 description 2
- SIKJAQJRHWYJAI-UHFFFAOYSA-N Indole Chemical compound C1=CC=C2NC=CC2=C1 SIKJAQJRHWYJAI-UHFFFAOYSA-N 0.000 description 2
- 239000004472 Lysine Substances 0.000 description 2
- AFVFQIVMOAPDHO-UHFFFAOYSA-N Methanesulfonic acid Chemical compound CS(O)(=O)=O AFVFQIVMOAPDHO-UHFFFAOYSA-N 0.000 description 2
- YNAVUWVOSKDBBP-UHFFFAOYSA-N Morpholine Chemical compound C1COCCN1 YNAVUWVOSKDBBP-UHFFFAOYSA-N 0.000 description 2
- PVNIIMVLHYAWGP-UHFFFAOYSA-N Niacin Chemical compound OC(=O)C1=CC=CN=C1 PVNIIMVLHYAWGP-UHFFFAOYSA-N 0.000 description 2
- GLUUGHFHXGJENI-UHFFFAOYSA-N Piperazine Chemical compound C1CNCCN1 GLUUGHFHXGJENI-UHFFFAOYSA-N 0.000 description 2
- QAOWNCQODCNURD-UHFFFAOYSA-N Sulfuric acid Chemical compound OS(O)(=O)=O QAOWNCQODCNURD-UHFFFAOYSA-N 0.000 description 2
- 239000003929 acidic solution Substances 0.000 description 2
- 239000004480 active ingredient Substances 0.000 description 2
- OIRDTQYFTABQOQ-KQYNXXCUSA-N adenosine Chemical compound C1=NC=2C(N)=NC=NC=2N1[C@@H]1O[C@H](CO)[C@@H](O)[C@H]1O OIRDTQYFTABQOQ-KQYNXXCUSA-N 0.000 description 2
- XAGFODPZIPBFFR-UHFFFAOYSA-N aluminium Chemical compound [Al] XAGFODPZIPBFFR-UHFFFAOYSA-N 0.000 description 2
- 229910052782 aluminium Inorganic materials 0.000 description 2
- 150000001408 amides Chemical group 0.000 description 2
- RDOXTESZEPMUJZ-UHFFFAOYSA-N anisole Chemical compound COC1=CC=CC=C1 RDOXTESZEPMUJZ-UHFFFAOYSA-N 0.000 description 2
- 230000003110 anti-inflammatory effect Effects 0.000 description 2
- 238000003556 assay Methods 0.000 description 2
- 125000001584 benzyloxycarbonyl group Chemical group C(=O)(OCC1=CC=CC=C1)* 0.000 description 2
- 230000015572 biosynthetic process Effects 0.000 description 2
- 239000002775 capsule Substances 0.000 description 2
- 210000004027 cell Anatomy 0.000 description 2
- 239000001913 cellulose Substances 0.000 description 2
- 235000010980 cellulose Nutrition 0.000 description 2
- 229920002678 cellulose Polymers 0.000 description 2
- 239000003153 chemical reaction reagent Substances 0.000 description 2
- 238000000354 decomposition reaction Methods 0.000 description 2
- AJDPNPAGZMZOMN-UHFFFAOYSA-N diethyl (4-oxo-1,2,3-benzotriazin-3-yl) phosphate Chemical compound C1=CC=C2C(=O)N(OP(=O)(OCC)OCC)N=NC2=C1 AJDPNPAGZMZOMN-UHFFFAOYSA-N 0.000 description 2
- HPNMFZURTQLUMO-UHFFFAOYSA-N diethylamine Chemical compound CCNCC HPNMFZURTQLUMO-UHFFFAOYSA-N 0.000 description 2
- JQCXWCOOWVGKMT-UHFFFAOYSA-N diheptyl phthalate Chemical compound CCCCCCCOC(=O)C1=CC=CC=C1C(=O)OCCCCCCC JQCXWCOOWVGKMT-UHFFFAOYSA-N 0.000 description 2
- 239000003814 drug Substances 0.000 description 2
- 239000000839 emulsion Substances 0.000 description 2
- 229940031098 ethanolamine Drugs 0.000 description 2
- 238000011156 evaluation Methods 0.000 description 2
- 210000002950 fibroblast Anatomy 0.000 description 2
- 239000006260 foam Substances 0.000 description 2
- 239000011888 foil Substances 0.000 description 2
- 235000013305 food Nutrition 0.000 description 2
- 238000009472 formulation Methods 0.000 description 2
- 239000000499 gel Substances 0.000 description 2
- 239000004220 glutamic acid Substances 0.000 description 2
- 239000008187 granular material Substances 0.000 description 2
- 239000001257 hydrogen Substances 0.000 description 2
- 229910052739 hydrogen Inorganic materials 0.000 description 2
- 239000004615 ingredient Substances 0.000 description 2
- 230000003834 intracellular effect Effects 0.000 description 2
- HQKMJHAJHXVSDF-UHFFFAOYSA-L magnesium stearate Chemical compound [Mg+2].CCCCCCCCCCCCCCCCCC([O-])=O.CCCCCCCCCCCCCCCCCC([O-])=O HQKMJHAJHXVSDF-UHFFFAOYSA-L 0.000 description 2
- 239000002609 medium Substances 0.000 description 2
- BDAGIHXWWSANSR-UHFFFAOYSA-N methanoic acid Natural products OC=O BDAGIHXWWSANSR-UHFFFAOYSA-N 0.000 description 2
- 239000008194 pharmaceutical composition Substances 0.000 description 2
- USHAGKDGDHPEEY-UHFFFAOYSA-L potassium persulfate Chemical compound [K+].[K+].[O-]S(=O)(=O)OOS([O-])(=O)=O USHAGKDGDHPEEY-UHFFFAOYSA-L 0.000 description 2
- 239000003755 preservative agent Substances 0.000 description 2
- 230000008569 process Effects 0.000 description 2
- 230000035755 proliferation Effects 0.000 description 2
- 239000008213 purified water Substances 0.000 description 2
- 239000000523 sample Substances 0.000 description 2
- 210000004761 scalp Anatomy 0.000 description 2
- 239000007921 spray Substances 0.000 description 2
- 238000006467 substitution reaction Methods 0.000 description 2
- 239000000725 suspension Substances 0.000 description 2
- 238000001308 synthesis method Methods 0.000 description 2
- 239000003826 tablet Substances 0.000 description 2
- 238000010998 test method 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
- AQRLNPVMDITEJU-UHFFFAOYSA-N triethylsilane Chemical compound CC[SiH](CC)CC AQRLNPVMDITEJU-UHFFFAOYSA-N 0.000 description 2
- UCPYLLCMEDAXFR-UHFFFAOYSA-N triphosgene Chemical compound ClC(Cl)(Cl)OC(=O)OC(Cl)(Cl)Cl UCPYLLCMEDAXFR-UHFFFAOYSA-N 0.000 description 2
- MBYLVOKEDDQJDY-UHFFFAOYSA-N tris(2-aminoethyl)amine Chemical compound NCCN(CCN)CCN MBYLVOKEDDQJDY-UHFFFAOYSA-N 0.000 description 2
- 239000011534 wash buffer Substances 0.000 description 2
- 125000003088 (fluoren-9-ylmethoxy)carbonyl group Chemical group 0.000 description 1
- WBYWAXJHAXSJNI-VOTSOKGWSA-M .beta-Phenylacrylic acid Natural products [O-]C(=O)\C=C\C1=CC=CC=C1 WBYWAXJHAXSJNI-VOTSOKGWSA-M 0.000 description 1
- OGJCIAFFKGRGJC-UHFFFAOYSA-N 1,2-bis(chloranyl)ethane Chemical compound ClCCCl.ClCCCl OGJCIAFFKGRGJC-UHFFFAOYSA-N 0.000 description 1
- BDNKZNFMNDZQMI-UHFFFAOYSA-N 1,3-diisopropylcarbodiimide Chemical compound CC(C)N=C=NC(C)C BDNKZNFMNDZQMI-UHFFFAOYSA-N 0.000 description 1
- FZTLLUYFWAOGGB-UHFFFAOYSA-N 1,4-dioxane dioxane Chemical compound C1COCCO1.C1COCCO1 FZTLLUYFWAOGGB-UHFFFAOYSA-N 0.000 description 1
- ASOKPJOREAFHNY-UHFFFAOYSA-N 1-Hydroxybenzotriazole Chemical compound C1=CC=C2N(O)N=NC2=C1 ASOKPJOREAFHNY-UHFFFAOYSA-N 0.000 description 1
- JFLSOKIMYBSASW-UHFFFAOYSA-N 1-chloro-2-[chloro(diphenyl)methyl]benzene Chemical compound ClC1=CC=CC=C1C(Cl)(C=1C=CC=CC=1)C1=CC=CC=C1 JFLSOKIMYBSASW-UHFFFAOYSA-N 0.000 description 1
- FPIRBHDGWMWJEP-UHFFFAOYSA-N 1-hydroxy-7-azabenzotriazole Chemical compound C1=CN=C2N(O)N=NC2=C1 FPIRBHDGWMWJEP-UHFFFAOYSA-N 0.000 description 1
- BSXPDVKSFWQFRT-UHFFFAOYSA-N 1-hydroxytriazolo[4,5-b]pyridine Chemical compound C1=CC=C2N(O)N=NC2=N1 BSXPDVKSFWQFRT-UHFFFAOYSA-N 0.000 description 1
- UAIUNKRWKOVEES-UHFFFAOYSA-N 3,3',5,5'-tetramethylbenzidine Chemical compound CC1=C(N)C(C)=CC(C=2C=C(C)C(N)=C(C)C=2)=C1 UAIUNKRWKOVEES-UHFFFAOYSA-N 0.000 description 1
- FPQQSJJWHUJYPU-UHFFFAOYSA-N 3-(dimethylamino)propyliminomethylidene-ethylazanium;chloride Chemical compound Cl.CCN=C=NCCCN(C)C FPQQSJJWHUJYPU-UHFFFAOYSA-N 0.000 description 1
- AGUDBAONKXJDNO-UHFFFAOYSA-N 3-[4-(3-aminopropyl)piperazin-1-yl]propan-1-amine Chemical compound NCCCN1CCN(CCCN)CC1.NCCCN1CCN(CCCN)CC1 AGUDBAONKXJDNO-UHFFFAOYSA-N 0.000 description 1
- VOUAQYXWVJDEQY-QENPJCQMSA-N 33017-11-7 Chemical compound OC(=O)CC[C@H](N)C(=O)N[C@@H](C)C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H](CC(O)=O)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CCC(N)=O)C(=O)N[C@@H](C(C)C)C(=O)NCC(=O)N[C@@H](CCC(N)=O)C(=O)N[C@@H](C(C)C)C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H](CC(C)C)C(=O)NCC(=O)NCC(=O)NCC(=O)N1CCC[C@H]1C(=O)NCC(=O)N[C@@H](C)C(=O)NCC(=O)N[C@@H](CO)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CCC(N)=O)C(=O)N1[C@H](C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](C)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CCC(O)=O)C(=O)NCC(=O)N[C@@H](CO)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CCC(N)=O)C(O)=O)CCC1 VOUAQYXWVJDEQY-QENPJCQMSA-N 0.000 description 1
- OSWFIVFLDKOXQC-UHFFFAOYSA-N 4-(3-methoxyphenyl)aniline Chemical compound COC1=CC=CC(C=2C=CC(N)=CC=2)=C1 OSWFIVFLDKOXQC-UHFFFAOYSA-N 0.000 description 1
- UZOFELREXGAFOI-UHFFFAOYSA-N 4-methylpiperidine Chemical compound CC1CCNCC1 UZOFELREXGAFOI-UHFFFAOYSA-N 0.000 description 1
- TZCYLJGNWDVJRA-UHFFFAOYSA-N 6-chloro-1-hydroxybenzotriazole Chemical compound C1=C(Cl)C=C2N(O)N=NC2=C1 TZCYLJGNWDVJRA-UHFFFAOYSA-N 0.000 description 1
- FHVDTGUDJYJELY-UHFFFAOYSA-N 6-{[2-carboxy-4,5-dihydroxy-6-(phosphanyloxy)oxan-3-yl]oxy}-4,5-dihydroxy-3-phosphanyloxane-2-carboxylic acid Chemical compound O1C(C(O)=O)C(P)C(O)C(O)C1OC1C(C(O)=O)OC(OP)C(O)C1O FHVDTGUDJYJELY-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
- 108010075254 C-Peptide Proteins 0.000 description 1
- 239000002126 C01EB10 - Adenosine Substances 0.000 description 1
- PTHCMJGKKRQCBF-UHFFFAOYSA-N Cellulose, microcrystalline Chemical compound OC1C(O)C(OC)OC(CO)C1OC1C(O)C(O)C(OC)C(CO)O1 PTHCMJGKKRQCBF-UHFFFAOYSA-N 0.000 description 1
- WBYWAXJHAXSJNI-SREVYHEPSA-N Cinnamic acid Chemical compound OC(=O)\C=C/C1=CC=CC=C1 WBYWAXJHAXSJNI-SREVYHEPSA-N 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
- FBPFZTCFMRRESA-KVTDHHQDSA-N D-Mannitol Chemical compound OC[C@@H](O)[C@@H](O)[C@H](O)[C@H](O)CO FBPFZTCFMRRESA-KVTDHHQDSA-N 0.000 description 1
- FBPFZTCFMRRESA-JGWLITMVSA-N D-glucitol Chemical compound OC[C@H](O)[C@@H](O)[C@H](O)[C@H](O)CO FBPFZTCFMRRESA-JGWLITMVSA-N 0.000 description 1
- 239000004375 Dextrin Substances 0.000 description 1
- 229920001353 Dextrin Polymers 0.000 description 1
- 235000011511 Diospyros Nutrition 0.000 description 1
- 244000236655 Diospyros kaki Species 0.000 description 1
- 238000011891 EIA kit Methods 0.000 description 1
- 239000004386 Erythritol Substances 0.000 description 1
- UNXHWFMMPAWVPI-UHFFFAOYSA-N Erythritol Natural products OCC(O)C(O)CO UNXHWFMMPAWVPI-UHFFFAOYSA-N 0.000 description 1
- 239000004606 Fillers/Extenders Substances 0.000 description 1
- 108010010803 Gelatin 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
- 229920002907 Guar gum Polymers 0.000 description 1
- 229920000084 Gum arabic Polymers 0.000 description 1
- 239000007821 HATU Substances 0.000 description 1
- 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 1
- 239000005913 Maltodextrin Substances 0.000 description 1
- 229920002774 Maltodextrin Polymers 0.000 description 1
- 229930195725 Mannitol Natural products 0.000 description 1
- 229920000168 Microcrystalline cellulose Polymers 0.000 description 1
- 239000004909 Moisturizer Substances 0.000 description 1
- ZKGNPQKYVKXMGJ-UHFFFAOYSA-N N,N-dimethylacetamide Chemical compound CN(C)C(C)=O.CN(C)C(C)=O ZKGNPQKYVKXMGJ-UHFFFAOYSA-N 0.000 description 1
- ZMXDDKWLCZADIW-UHFFFAOYSA-N N,N-dimethylformamide Substances CN(C)C=O ZMXDDKWLCZADIW-UHFFFAOYSA-N 0.000 description 1
- NQTADLQHYWFPDB-UHFFFAOYSA-N N-Hydroxysuccinimide Chemical compound ON1C(=O)CCC1=O NQTADLQHYWFPDB-UHFFFAOYSA-N 0.000 description 1
- 102000003992 Peroxidases Human genes 0.000 description 1
- 239000004793 Polystyrene Substances 0.000 description 1
- 230000002292 Radical scavenging effect Effects 0.000 description 1
- 229920002472 Starch Polymers 0.000 description 1
- 229930006000 Sucrose Natural products 0.000 description 1
- CZMRCDWAGMRECN-UGDNZRGBSA-N Sucrose Chemical compound O[C@H]1[C@H](O)[C@@H](CO)O[C@@]1(CO)O[C@@H]1[C@H](O)[C@@H](O)[C@H](O)[C@@H](CO)O1 CZMRCDWAGMRECN-UGDNZRGBSA-N 0.000 description 1
- 239000012317 TBTU Substances 0.000 description 1
- 241000289690 Xenarthra Species 0.000 description 1
- TVXBFESIOXBWNM-UHFFFAOYSA-N Xylitol Natural products OCCC(O)C(O)C(O)CCO TVXBFESIOXBWNM-UHFFFAOYSA-N 0.000 description 1
- RDWDVLFMPFUBDV-PXMDEAMVSA-N [(e)-(1-cyano-2-ethoxy-2-oxoethylidene)amino]oxy-tripyrrolidin-1-ylphosphanium;hexafluorophosphate Chemical compound F[P-](F)(F)(F)(F)F.C1CCCN1[P+](N1CCCC1)(O/N=C(C(=O)OCC)\C#N)N1CCCC1 RDWDVLFMPFUBDV-PXMDEAMVSA-N 0.000 description 1
- NERFNHBZJXXFGY-UHFFFAOYSA-N [4-[(4-methylphenyl)methoxy]phenyl]methanol Chemical compound C1=CC(C)=CC=C1COC1=CC=C(CO)C=C1 NERFNHBZJXXFGY-UHFFFAOYSA-N 0.000 description 1
- GPDHNZNLPKYHCN-DZOOLQPHSA-N [[(z)-(1-cyano-2-ethoxy-2-oxoethylidene)amino]oxy-morpholin-4-ylmethylidene]-dimethylazanium;hexafluorophosphate Chemical compound F[P-](F)(F)(F)(F)F.CCOC(=O)C(\C#N)=N/OC(=[N+](C)C)N1CCOCC1 GPDHNZNLPKYHCN-DZOOLQPHSA-N 0.000 description 1
- CLZISMQKJZCZDN-UHFFFAOYSA-N [benzotriazol-1-yloxy(dimethylamino)methylidene]-dimethylazanium Chemical compound C1=CC=C2N(OC(N(C)C)=[N+](C)C)N=NC2=C1 CLZISMQKJZCZDN-UHFFFAOYSA-N 0.000 description 1
- ITLHXEGAYQFOHJ-UHFFFAOYSA-N [diazo(phenyl)methyl]benzene Chemical compound C=1C=CC=CC=1C(=[N+]=[N-])C1=CC=CC=C1 ITLHXEGAYQFOHJ-UHFFFAOYSA-N 0.000 description 1
- 235000010489 acacia gum Nutrition 0.000 description 1
- 239000000205 acacia gum Substances 0.000 description 1
- 125000002777 acetyl group Chemical group [H]C([H])([H])C(*)=O 0.000 description 1
- 239000002253 acid Substances 0.000 description 1
- 230000009471 action Effects 0.000 description 1
- 229960005305 adenosine Drugs 0.000 description 1
- 239000000443 aerosol Substances 0.000 description 1
- 229940072056 alginate Drugs 0.000 description 1
- 235000010443 alginic acid Nutrition 0.000 description 1
- 229920000615 alginic acid Polymers 0.000 description 1
- 125000000539 amino acid group Chemical group 0.000 description 1
- 125000003277 amino group Chemical group 0.000 description 1
- 238000004458 analytical method Methods 0.000 description 1
- 230000000844 anti-bacterial effect Effects 0.000 description 1
- 230000003367 anti-collagen effect Effects 0.000 description 1
- 150000004982 aromatic amines Chemical group 0.000 description 1
- OHDRQQURAXLVGJ-AXMZSLBLSA-N azane;(2z)-3-ethyl-2-[(z)-(3-ethyl-6-sulfo-1,3-benzothiazol-2-ylidene)hydrazinylidene]-1,3-benzothiazole-6-sulfonic acid Chemical compound [NH4+].[NH4+].S/1C2=CC(S([O-])(=O)=O)=CC=C2N(CC)C\1=N\N=C1/SC2=CC(S([O-])(=O)=O)=CC=C2N1CC OHDRQQURAXLVGJ-AXMZSLBLSA-N 0.000 description 1
- 239000003899 bactericide agent Substances 0.000 description 1
- 125000003236 benzoyl group Chemical group [H]C1=C([H])C([H])=C(C([H])=C1[H])C(*)=O 0.000 description 1
- WQZGKKKJIJFFOK-VFUOTHLCSA-N beta-D-glucose Chemical compound OC[C@H]1O[C@@H](O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-VFUOTHLCSA-N 0.000 description 1
- 239000011230 binding agent Substances 0.000 description 1
- 230000033228 biological regulation Effects 0.000 description 1
- 239000012496 blank sample Substances 0.000 description 1
- 230000000903 blocking effect Effects 0.000 description 1
- 239000000872 buffer Substances 0.000 description 1
- HTRXGEPDTFSKLI-UHFFFAOYSA-N butanoic acid;ethyl acetate Chemical compound CCCC(O)=O.CCOC(C)=O HTRXGEPDTFSKLI-UHFFFAOYSA-N 0.000 description 1
- 210000001217 buttock Anatomy 0.000 description 1
- FNAQSUUGMSOBHW-UHFFFAOYSA-H calcium citrate Chemical compound [Ca+2].[Ca+2].[Ca+2].[O-]C(=O)CC(O)(CC([O-])=O)C([O-])=O.[O-]C(=O)CC(O)(CC([O-])=O)C([O-])=O FNAQSUUGMSOBHW-UHFFFAOYSA-H 0.000 description 1
- 239000001354 calcium citrate Substances 0.000 description 1
- AXCZMVOFGPJBDE-UHFFFAOYSA-L calcium dihydroxide Chemical compound [OH-].[OH-].[Ca+2] AXCZMVOFGPJBDE-UHFFFAOYSA-L 0.000 description 1
- 239000000920 calcium hydroxide Substances 0.000 description 1
- 229910001861 calcium hydroxide Inorganic materials 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
- 235000012241 calcium silicate Nutrition 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
- 150000001732 carboxylic acid derivatives Chemical class 0.000 description 1
- 239000000969 carrier Substances 0.000 description 1
- 230000015556 catabolic process Effects 0.000 description 1
- 238000004113 cell culture Methods 0.000 description 1
- 229930016911 cinnamic acid Natural products 0.000 description 1
- 235000013985 cinnamic acid Nutrition 0.000 description 1
- 230000000052 comparative effect Effects 0.000 description 1
- 150000001875 compounds Chemical class 0.000 description 1
- 239000000470 constituent Substances 0.000 description 1
- 238000007796 conventional method Methods 0.000 description 1
- 239000011162 core material Substances 0.000 description 1
- XUJNEKJLAYXESH-UHFFFAOYSA-N cysteine Natural products SCC(N)C(O)=O XUJNEKJLAYXESH-UHFFFAOYSA-N 0.000 description 1
- 235000018417 cysteine Nutrition 0.000 description 1
- VTXVGVNLYGSIAR-UHFFFAOYSA-N decane-1-thiol Chemical compound CCCCCCCCCCS VTXVGVNLYGSIAR-UHFFFAOYSA-N 0.000 description 1
- 230000007423 decrease Effects 0.000 description 1
- 238000010511 deprotection reaction Methods 0.000 description 1
- 238000011161 development Methods 0.000 description 1
- 230000018109 developmental process Effects 0.000 description 1
- 235000019425 dextrin Nutrition 0.000 description 1
- 239000008121 dextrose Substances 0.000 description 1
- 238000007865 diluting Methods 0.000 description 1
- UXGNZZKBCMGWAZ-UHFFFAOYSA-N dimethylformamide dmf Chemical compound CN(C)C=O.CN(C)C=O UXGNZZKBCMGWAZ-UHFFFAOYSA-N 0.000 description 1
- 239000007884 disintegrant Substances 0.000 description 1
- PXEDJBXQKAGXNJ-QTNFYWBSSA-L disodium L-glutamate Chemical compound [Na+].[Na+].[O-]C(=O)[C@@H](N)CCC([O-])=O PXEDJBXQKAGXNJ-QTNFYWBSSA-L 0.000 description 1
- 239000012153 distilled water Substances 0.000 description 1
- 229940079593 drug Drugs 0.000 description 1
- 239000000975 dye Substances 0.000 description 1
- 235000019414 erythritol Nutrition 0.000 description 1
- UNXHWFMMPAWVPI-ZXZARUISSA-N erythritol Chemical compound OC[C@H](O)[C@H](O)CO UNXHWFMMPAWVPI-ZXZARUISSA-N 0.000 description 1
- 229940009714 erythritol Drugs 0.000 description 1
- GVXHLXUFBFUGKE-UHFFFAOYSA-N ethane-1,2-dithiol Chemical compound SCCS.SCCS GVXHLXUFBFUGKE-UHFFFAOYSA-N 0.000 description 1
- 230000001747 exhibiting effect Effects 0.000 description 1
- 239000000945 filler Substances 0.000 description 1
- 239000000706 filtrate Substances 0.000 description 1
- 239000012467 final product Substances 0.000 description 1
- 230000037406 food intake Effects 0.000 description 1
- 235000019253 formic acid Nutrition 0.000 description 1
- 239000003205 fragrance Substances 0.000 description 1
- 239000012737 fresh medium Substances 0.000 description 1
- 125000000524 functional group Chemical group 0.000 description 1
- 239000008273 gelatin Substances 0.000 description 1
- 229920000159 gelatin Polymers 0.000 description 1
- 235000019322 gelatine Nutrition 0.000 description 1
- 235000011852 gelatine desserts Nutrition 0.000 description 1
- 238000012812 general test Methods 0.000 description 1
- 239000000665 guar gum Substances 0.000 description 1
- 235000010417 guar gum Nutrition 0.000 description 1
- 229960002154 guar gum Drugs 0.000 description 1
- 239000000118 hair dye Substances 0.000 description 1
- HNDVDQJCIGZPNO-UHFFFAOYSA-N histidine Natural products OC(=O)C(N)CC1=CN=CN1 HNDVDQJCIGZPNO-UHFFFAOYSA-N 0.000 description 1
- 125000002887 hydroxy group Chemical group [H]O* 0.000 description 1
- NPZTUJOABDZTLV-UHFFFAOYSA-N hydroxybenzotriazole Substances O=C1C=CC=C2NNN=C12 NPZTUJOABDZTLV-UHFFFAOYSA-N 0.000 description 1
- 230000008676 import Effects 0.000 description 1
- 238000011534 incubation Methods 0.000 description 1
- PZOUSPYUWWUPPK-UHFFFAOYSA-N indole Natural products CC1=CC=CC2=C1C=CN2 PZOUSPYUWWUPPK-UHFFFAOYSA-N 0.000 description 1
- RKJUIXBNRJVNHR-UHFFFAOYSA-N indolenine Natural products C1=CC=C2CC=NC2=C1 RKJUIXBNRJVNHR-UHFFFAOYSA-N 0.000 description 1
- 238000002347 injection Methods 0.000 description 1
- 239000007924 injection Substances 0.000 description 1
- 150000007529 inorganic bases Chemical class 0.000 description 1
- 210000002510 keratinocyte Anatomy 0.000 description 1
- 150000002576 ketones Chemical class 0.000 description 1
- 239000008101 lactose Substances 0.000 description 1
- 229940069445 licorice extract Drugs 0.000 description 1
- 235000019359 magnesium stearate Nutrition 0.000 description 1
- 239000000845 maltitol Substances 0.000 description 1
- 235000010449 maltitol Nutrition 0.000 description 1
- VQHSOMBJVWLPSR-WUJBLJFYSA-N maltitol Chemical compound OC[C@H](O)[C@@H](O)[C@@H]([C@H](O)CO)O[C@H]1O[C@H](CO)[C@@H](O)[C@H](O)[C@H]1O VQHSOMBJVWLPSR-WUJBLJFYSA-N 0.000 description 1
- 229940035436 maltitol Drugs 0.000 description 1
- 229940035034 maltodextrin Drugs 0.000 description 1
- 239000000594 mannitol Substances 0.000 description 1
- 235000010355 mannitol Nutrition 0.000 description 1
- 229960001855 mannitol Drugs 0.000 description 1
- 238000005259 measurement Methods 0.000 description 1
- 239000012528 membrane Substances 0.000 description 1
- HEBKCHPVOIAQTA-UHFFFAOYSA-N meso ribitol Natural products OCC(O)C(O)C(O)CO HEBKCHPVOIAQTA-UHFFFAOYSA-N 0.000 description 1
- 229940098779 methanesulfonic acid Drugs 0.000 description 1
- UZKWTJUDCOPSNM-UHFFFAOYSA-N methoxybenzene Substances CCCCOC=C UZKWTJUDCOPSNM-UHFFFAOYSA-N 0.000 description 1
- RCYSGAYIEFAJTG-UHFFFAOYSA-N methyl acetate;propanoic acid Chemical compound CCC(O)=O.COC(C)=O RCYSGAYIEFAJTG-UHFFFAOYSA-N 0.000 description 1
- 229920000609 methyl cellulose Polymers 0.000 description 1
- WBYWAXJHAXSJNI-UHFFFAOYSA-N methyl p-hydroxycinnamate Natural products OC(=O)C=CC1=CC=CC=C1 WBYWAXJHAXSJNI-UHFFFAOYSA-N 0.000 description 1
- 239000001923 methylcellulose Substances 0.000 description 1
- 235000010981 methylcellulose Nutrition 0.000 description 1
- LXCFILQKKLGQFO-UHFFFAOYSA-N methylparaben Chemical compound COC(=O)C1=CC=C(O)C=C1 LXCFILQKKLGQFO-UHFFFAOYSA-N 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
- 239000002480 mineral oil Substances 0.000 description 1
- 235000010446 mineral oil Nutrition 0.000 description 1
- 239000011259 mixed solution Substances 0.000 description 1
- 238000012986 modification Methods 0.000 description 1
- 230000004048 modification Effects 0.000 description 1
- 230000001333 moisturizer Effects 0.000 description 1
- 239000002324 mouth wash Substances 0.000 description 1
- PEECTLLHENGOKU-UHFFFAOYSA-N n,n-dimethylpyridin-4-amine Chemical compound CN(C)C1=CC=NC=C1.CN(C)C1=CC=NC=C1 PEECTLLHENGOKU-UHFFFAOYSA-N 0.000 description 1
- 235000019462 natural additive Nutrition 0.000 description 1
- 239000011664 nicotinic acid Substances 0.000 description 1
- 235000001968 nicotinic acid Nutrition 0.000 description 1
- 229960003512 nicotinic acid Drugs 0.000 description 1
- 150000002825 nitriles Chemical class 0.000 description 1
- VWBWQOUWDOULQN-UHFFFAOYSA-N nmp n-methylpyrrolidone Chemical compound CN1CCCC1=O.CN1CCCC1=O VWBWQOUWDOULQN-UHFFFAOYSA-N 0.000 description 1
- 231100000252 nontoxic Toxicity 0.000 description 1
- 230000003000 nontoxic effect Effects 0.000 description 1
- 235000015097 nutrients Nutrition 0.000 description 1
- 239000002674 ointment Substances 0.000 description 1
- 239000006186 oral dosage form Substances 0.000 description 1
- 150000007530 organic bases Chemical class 0.000 description 1
- 238000010647 peptide synthesis reaction Methods 0.000 description 1
- 108040007629 peroxidase activity proteins Proteins 0.000 description 1
- 229940044654 phenolsulfonic acid Drugs 0.000 description 1
- 239000008363 phosphate buffer Substances 0.000 description 1
- 239000006187 pill Substances 0.000 description 1
- 229920002223 polystyrene Polymers 0.000 description 1
- 239000001267 polyvinylpyrrolidone Substances 0.000 description 1
- 235000013855 polyvinylpyrrolidone Nutrition 0.000 description 1
- 229920000036 polyvinylpyrrolidone Polymers 0.000 description 1
- ATLPLEZDTSBZQG-UHFFFAOYSA-N propan-2-ylphosphonic acid Chemical compound CC(C)P(O)(O)=O ATLPLEZDTSBZQG-UHFFFAOYSA-N 0.000 description 1
- QELSKZZBTMNZEB-UHFFFAOYSA-N propylparaben Chemical compound CCCOC(=O)C1=CC=C(O)C=C1 QELSKZZBTMNZEB-UHFFFAOYSA-N 0.000 description 1
- 229960003415 propylparaben Drugs 0.000 description 1
- 238000000746 purification Methods 0.000 description 1
- 150000003839 salts Chemical class 0.000 description 1
- 239000012488 sample solution Substances 0.000 description 1
- 238000000926 separation method Methods 0.000 description 1
- 239000012679 serum free medium Substances 0.000 description 1
- 239000002453 shampoo Substances 0.000 description 1
- 210000004927 skin cell Anatomy 0.000 description 1
- 239000007790 solid phase Substances 0.000 description 1
- 238000010532 solid phase synthesis reaction Methods 0.000 description 1
- 235000010356 sorbitol Nutrition 0.000 description 1
- 239000000600 sorbitol Substances 0.000 description 1
- 229960002920 sorbitol Drugs 0.000 description 1
- 239000012086 standard solution Substances 0.000 description 1
- 239000008107 starch Substances 0.000 description 1
- 229940032147 starch Drugs 0.000 description 1
- 235000019698 starch Nutrition 0.000 description 1
- 239000005720 sucrose Substances 0.000 description 1
- 230000000475 sunscreen effect Effects 0.000 description 1
- 239000000516 sunscreening agent Substances 0.000 description 1
- 239000006228 supernatant Substances 0.000 description 1
- 239000000829 suppository Substances 0.000 description 1
- 239000004094 surface-active agent Substances 0.000 description 1
- 230000008961 swelling Effects 0.000 description 1
- 230000002194 synthesizing effect Effects 0.000 description 1
- 229920001059 synthetic polymer Polymers 0.000 description 1
- 239000006188 syrup Substances 0.000 description 1
- 235000020357 syrup Nutrition 0.000 description 1
- 239000000454 talc Substances 0.000 description 1
- 229910052623 talc Inorganic materials 0.000 description 1
- 235000012222 talc Nutrition 0.000 description 1
- 239000002562 thickening agent Substances 0.000 description 1
- 239000000606 toothpaste Substances 0.000 description 1
- 230000000699 topical effect 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
- 235000013337 tricalcium citrate Nutrition 0.000 description 1
- 239000000080 wetting agent Substances 0.000 description 1
- 230000002087 whitening effect Effects 0.000 description 1
- 239000000811 xylitol Substances 0.000 description 1
- 235000010447 xylitol Nutrition 0.000 description 1
- HEBKCHPVOIAQTA-SCDXWVJYSA-N xylitol Chemical compound OC[C@H](O)[C@@H](O)[C@H](O)CO HEBKCHPVOIAQTA-SCDXWVJYSA-N 0.000 description 1
- 229960002675 xylitol Drugs 0.000 description 1
Images
Classifications
-
- 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
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
- A23L33/00—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof
- A23L33/10—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof using additives
- A23L33/17—Amino acids, peptides or proteins
-
- 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
- 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
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P17/00—Drugs for dermatological disorders
- A61P17/18—Antioxidants, e.g. antiradicals
-
- 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
- A61Q19/02—Preparations for care of the skin for chemically bleaching or whitening the skin
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
- A23V2002/00—Food compositions, function of food ingredients or processes for food or foodstuffs
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
- A23V2200/00—Function of food ingredients
- A23V2200/02—Antioxidant
Landscapes
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Chemical & Material Sciences (AREA)
- General Health & Medical Sciences (AREA)
- Public Health (AREA)
- Animal Behavior & Ethology (AREA)
- Veterinary Medicine (AREA)
- Organic Chemistry (AREA)
- Engineering & Computer Science (AREA)
- Medicinal Chemistry (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- Epidemiology (AREA)
- Pharmacology & Pharmacy (AREA)
- Dermatology (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Biochemistry (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Biophysics (AREA)
- Genetics & Genomics (AREA)
- Molecular Biology (AREA)
- Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
- Birds (AREA)
- General Chemical & Material Sciences (AREA)
- Mycology (AREA)
- Nutrition Science (AREA)
- Food Science & Technology (AREA)
- Polymers & Plastics (AREA)
- Gastroenterology & Hepatology (AREA)
- Immunology (AREA)
- Peptides Or Proteins (AREA)
- Medicines That Contain Protein Lipid Enzymes And Other Medicines (AREA)
Abstract
본 발명은 항산화 활성 및 콜라게나제 저해활성을 갖는 펩타이드 및 이를 포함하는 조성물에 관한 것이다.
본 발명에 따른 펩타이드는 하기 화학식 I의 구조를 갖는다.
[화학식 I]
본 발명의 신규한 펩타이드는 항산화 활성 및 콜라게나제 저해활성이 우수하여 주름 개선, 노화 방지 효과가 뛰어나므로, 인체 적용시 본래의 기능을 효과적으로 발휘할 수 있어, 약학, 화장품, 기능성 식품 분야에서 유용하게 이용될 수 있다.The present invention relates to a peptide having antioxidant activity and collagenase inhibitory activity, and a composition comprising the same.
The peptide according to the present invention has the structure of the following formula (I).
[Formula I]
The novel peptide of the present invention has excellent antioxidant activity and collagenase inhibitory activity, so it has excellent anti-wrinkle and anti-aging effects. can be used
Description
본 발명은 항산화 활성 및 콜라게나제 저해활성을 갖는 펩타이드 및 이를 포함하는 조성물에 관한 것이다.The present invention relates to a peptide having antioxidant activity and collagenase inhibitory activity, and a composition comprising the same.
펩타이드는 바이오의 핵심소재로 단백질의 기능적 최소 단위이며, 아미노산이 2개로부터 50개 이하로 구성된다. 소량으로 우수한 효능을 보이면서 독성이 없어 의약품, 식품 및 화장품의 주요 원료로 많이 활용되고 있다. 특히 인체에 안전하고 효과가 좋은 펩타이드 화장품 소재가 수요가 증가함에 따라 신규 소재의 개발은 수입대체 효과 등 당 산업에서 매우 중요하다고 할 수 있다.Peptides are the core material of bio, the functional minimum unit of protein, and consist of 2 to 50 amino acids. It shows excellent efficacy in a small amount and is non-toxic, so it is widely used as a major raw material for pharmaceuticals, food and cosmetics. In particular, as the demand for peptide cosmetic materials that are safe and effective for the human body increases, the development of new materials is very important in the industry, such as import substitution effect.
화장품 원료로 사용하는 펩타이드의 효능은 주로 콜라겐 증식, 미백, 항염 등의 효능을 단독으로 가지는 소재가 주를 이루고 있으며, 비교적 고가 소재이기 때문에 사용량에 제한이 있는 실정이다. 이러한 항산화, 항염, 콜라겐 증식 등 다양한 효능을 하나의 펩타이드를 통해 발현할 수 있을 뿐만 아니라 비교적 짧은 서열로 구성되어 대량 생산을 통해 상업화가 가능하다면 관련 분야에서 다양하게 활용 가능하다. The efficacy of peptides used as cosmetic raw materials is mainly made up of materials that have effects such as collagen proliferation, whitening, and anti-inflammatory properties alone, and since they are relatively expensive materials, there is a limit to the amount used. If various effects such as antioxidant, anti-inflammatory, and collagen proliferation can be expressed through a single peptide, and it is composed of a relatively short sequence and can be commercialized through mass production, it can be used in various fields in related fields.
콜라겐은 진피에 위치하며 합성과 분해를 반복하면서, 새로운 콜라겐으로 교체된다. 콜라겐은 피부에 있어 매우 중요한 단백질로 진피 무게의 70-80%를 차지하는데 이 때문에 피부의 주름은 콜라겐의 합성과 분해의 불균형에 기인한다고 볼수 있다. 우리 피부가 강한 자외선에 무방비로 노출되면 피부세포에 콜라겐 분해효소(Collagenase)가 많이 만들어져 콜라겐의 분해가 지나치게 빠른 속도로 진행된다. 그러면 합성 속도가 이를 따라가지 못해 진피의 콜라겐 양이 감소하면서 피부의 탄력이 떨어져 주름살의 원인이 된다. Collagen is located in the dermis and is replaced with new collagen while repeating synthesis and decomposition. Collagen is a very important protein for the skin and accounts for 70-80% of the weight of the dermis. When our skin is exposed to strong UV rays unprotected, a lot of collagenase is produced in the skin cells, and the breakdown of collagen proceeds too quickly. Then, the synthesis rate cannot keep up with it, and the amount of collagen in the dermis decreases, causing the skin to lose elasticity and cause wrinkles.
한국공개특허 제2019-0128603호는 주름개선 효능을 갖는 펩티드를 개시하였고, 한국등록특허 제1957352호 및 제1957352호는 주름 개선 활성을 나타내는 펩타이드를 개시하였으나, 항산화 및 콜라게나제 저해 효능을 동시에 갖지는 않으므로, 복수의 생리활성 효과를 갖는 펩타이드의 개발이 절실한 상황이다.Korean Patent Application Laid-Open No. 2019-0128603 discloses a peptide having anti-wrinkle effect, and Korean Patent No. 1957352 and 1957352 disclose a peptide exhibiting wrinkle-improving activity, but have antioxidant and collagenase inhibitory effects at the same time. Therefore, there is an urgent need to develop peptides having multiple physiologically active effects.
이에, 본 발명자들은 이와 같은 콜라겐 분해활성을 저해함으로서 주름방지 효과를 가지는 펩타이드 서열을 확인한 결과 항산화와 콜라게나제 저해활성이 우수함을 알아내어, 본 발명을 완성하게 되었다.Accordingly, the present inventors confirmed the peptide sequence having an anti-wrinkle effect by inhibiting the collagen decomposition activity, and found that it has excellent antioxidant and collagenase inhibitory activity, and completed the present invention.
본 발명의 목적은 항산화 활성 및 콜라게나제 저해활성이 우수한 신규 펩타이드 및 이의 용도를 제공하는 데 있다.It is an object of the present invention to provide a novel peptide having excellent antioxidant activity and collagenase inhibitory activity, and uses thereof.
본 발명의 다른 목적은 주름 개선 및 피부 노화 방지용 조성물을 제공하는 데 있다.Another object of the present invention is to provide a composition for improving wrinkles and preventing skin aging.
본 발명자들은 상기에 언급한 바와 같이 신규 펩타이드를 설계하고 제조하여 효능을 확인한 결과 항산화 활성 및 콜라게나제 저해활성이 우수함을 확인함으로써 본 발명을 완성하였다.As mentioned above, the present inventors completed the present invention by designing and manufacturing a novel peptide and confirming the efficacy of the novel peptide by confirming that it has excellent antioxidant activity and collagenase inhibitory activity.
본 발명은 항산화 활성 및 콜라게나제 저해활성을 갖는 하기 화학식 I로 표시되는 펩타이드를 제공하는 것을 목적으로 한다.An object of the present invention is to provide a peptide represented by the following formula (I) having antioxidant activity and collagenase inhibitory activity.
[화학식 I][Formula I]
상기 화학식 I의 펩타이드는 H-γ-Glu-Cys-γ-Glu-Cys-Gly-His-Lys-OH의 서열을 가진 헵타펩타이드(Heptapeptide) 이다.The peptide of Formula I is a heptapeptide having a sequence of H-γ-Glu-Cys-γ-Glu-Cys-Gly-His-Lys-OH.
본 발명에 있어서, 상기 화학식 I의 신규 펩타이드는 당 업계에 알려진 기술을 사용하여 제조할 수 있다. 대표적인 합성방법을 아래에 설명하고 있으나, 본 발명이 이러한 예에 제한되는 것은 아니며, 당 업계에 알려진 기술의 범위 내에서 당 업자가 적절히 변형하여 사용할 수 있다.In the present invention, the novel peptide of Formula I can be prepared using techniques known in the art. Representative synthesis methods are described below, but the present invention is not limited to these examples, and those skilled in the art can use it with appropriate modifications within the range of techniques known in the art.
상기 목적을 달성하기 위하여, 본 발명은 (a) H-Lys(Boc)-Wang 레진에 Fmoc-His(Trt)-OH를 결합하여 화학식 I-a로 표시되는 Fmoc-His(Trt)-Lys(Boc)-Wang 레진을 얻는 단계; (b) 상기 단계 (a)에서 수득한 레진에 아미노산 (Fmoc-Gly-OH, Fmoc-Cys(Trt)-OH, Fmoc-γ-Glu-OtBu 등)을 순서적으로 결합하여 화학식 I-b로 표시되는 보호기를 가진 레진에 결합된 펩타이드를 얻는 단계; (c) 상기 단계 (b)에서 수득한 보호화된 펩타이드가 결합된 레진으로부터 탈 보호화 공정을 거쳐 화학식 I-c로 표시되는 탈보호화된 Crude 펩타이드를 수득하는 단계; 및 (d) 상기 단계 (c)에서 수득한 비 정제 (Crude)된 펩타이드를 고성능액체크로마토그래피 (HPLC)로 정제하여 화학식 I로 표시되는 펩타이드를 얻는 단계를 포함하는 펩타이드의 제조방법을 제공한다.In order to achieve the above object, the present invention provides (a) Fmoc-His(Trt)-Lys(Boc) represented by Formula I-a by binding Fmoc-His(Trt)-OH to H-Lys(Boc)-Wang resin. - Obtaining Wang resin; (b) amino acids (Fmoc-Gly-OH, Fmoc-Cys(Trt)-OH, Fmoc-γ-Glu-OtBu, etc.) obtaining a peptide bound to a resin having a protecting group; (c) obtaining a deprotected crude peptide represented by Formula I-c through a deprotection process from the resin to which the protected peptide obtained in step (b) is bound; and (d) purifying the crude peptide obtained in step (c) by high performance liquid chromatography (HPLC) to obtain a peptide represented by Formula I.
[화학식 I-a][Formula I-a]
[화학식 I-b][Formula I-b]
[화학식 I-c][Formula I-c]
[화학식 I][Formula I]
본 발명에 있어서, 상기 화학식 I로 표시되는 펩타이드는 고체상 합성법으로 합성하는 것을 특징으로 한다.In the present invention, the peptide represented by the formula (I) is characterized in that it is synthesized by a solid-phase synthesis method.
상기 화학식 I-a 및 I-b에서 R₁은 당 업계에서 통상적으로 이용하는 수소 또는 아민 보호기를 이용할 수 있다. 상기 아민 보호기는 아세트아미노메틸 (Acetaminomethyl)기, 벤질옥시카보닐 (Benzyloxycarbonyl)기, 터트-부틸옥시카르보닐 (tert-Butyloxycarbonyl)기, 아세틸 (acetyl)기, 벤조일 (Benzoyl)기, 파라-니트로벤조일 (para-Nitrobenzoyl)기, 파라-메톡시벤조일기 (para-Methoxybenzoyl) 등을 예시할 수 있으며, 벤질옥시카보닐 (Benzyloxycarbonyl)기, 터트-부틸옥시카르보닐 (tert-Butyloxycarbonyl)기가 바람직하며, 터트-부틸옥시카르보닐 (tert-Butyloxycarbonyl)기를 이용하는 것이 보다 바람직하지만 이에 국한되는 것은 아니다.In Formulas Ia and Ib, R₁ may use hydrogen or an amine protecting group commonly used in the art. The amine protecting group is an acetaminomethyl group, a benzyloxycarbonyl group, a tert -butyloxycarbonyl group, an acetyl group, a benzoyl group, and a para-nitrobenzoyl group. ( para -Nitrobenzoyl) group, para-methoxybenzoyl group ( para -Methoxybenzoyl), etc. can be exemplified, and a benzyloxycarbonyl group, a tert -butyloxycarbonyl group is preferable, and tert -Butyloxycarbonyl ( tert -Butyloxycarbonyl) is more preferable to use a group, but is not limited thereto.
상기 화학식 I-b에서 R₂는 당 업계에서 통상적으로 이용하는 수소 또는 방향족 아민 보호기를 이용할 수 있다. 상기 아민 보호기는 메톡시메틸 (Methoxymethyl)기, 벤질옥시메틸 (Benzyloxymethyl)기, 트리페닐메틸 (Triphenylmethyl)기, 터트-부틸디메틸실릴 (tert-Butyldimethylsilyl)기, 트리페닐실릴 (Triphenylsily)기, 트리이소프로필실릴 (Triispropylsilyl)기, 파라-메톡시벤질 (para-Methylxybenzyl)기, 테트라히드로피란 (Tetrahydropyran)기, 테트라히드로퓨란 (Tetrahydrofuran)기, 터트-부틸 (tert-Butyl)기, 디페닐메틸 (Diphenylmethyl)기, 2-클로로트리틸 (2-Chlorotrityl)기, 벤질 (Benzyl)기, 4-메톡시벤질 (4-Methoxybenzyl)기, 알릴 (Allyl)기, 터트-부틸옥시카르보닐 (tert-Butyloxycarbonyl)기, 아세틸 (Acetyl)기, 벤조일 (Benzoyl)기 등을 예시할 수 있으며, 메톡시메틸 (Methoxymethyl)기, 벤질옥시메틸 (Benzyloxymethyl)기, 트리페닐메틸 (Triphenylmethyl)기, 터트-부틸디메틸실릴 (tert-Butyldimethylsilyl)기, 트리페닐실릴 (Triphenylsily)기 또는 트리이소프로필실릴 (Triisopropylsilyl)기를 이용하는 것이 바람직하며, 2-클로로트리틸 (2-Chlorotrityl)기 또는 트리페닐메틸 (Triphenylmethyl)기를 이용하는 것이 보다 바람직하며, 아미드 보호기가 트리페닐메틸 (Triphenylmethyl)기를 이용하는 것이 가장 바람직하지만 이에 국한되는 것은 아니다.In Formula Ib, R2 may use hydrogen or an aromatic amine protecting group commonly used in the art. The amine protecting group is a methoxymethyl group, a benzyloxymethyl group, a triphenylmethyl group, a tert -butyldimethylsilyl group, a triphenylsily group, and a triiso group. Triispropylsilyl group, para-methoxybenzyl ( para -Methylxybenzyl) group, tetrahydropyran (Tetrahydropyran) group, tetrahydrofuran (Tetrahydrofuran) group, tert-butyl ( tert -Butyl) group, diphenylmethyl (Diphenylmethyl) ) group, 2-chlorotrityl (2-Chlorotrityl) group, benzyl (Benzyl) group, 4-methoxybenzyl (4-Methoxybenzyl) group, allyl (Allyl) group, tert -Butyloxycarbonyl group, acetyl (Acetyl) group, benzoyl (Benzoyl) group, etc. can be exemplified, and methoxymethyl (Methoxymethyl) group, benzyloxymethyl (Benzyloxymethyl) group, triphenylmethyl (Triphenylmethyl) group, tert-butyldimethylsilyl ( It is preferable to use a tert -Butyldimethylsilyl) group, a triphenylsily group, or a triisopropylsilyl group, and it is more preferable to use a 2-chlorotrityl (2-Chlorotrityl) group or a triphenylmethyl group. and it is most preferable to use a triphenylmethyl group as the amide protecting group, but is not limited thereto.
상기 화학식 I-b에서 R₃은 당 업계에서 통상적으로 이용하는 티올 보호기를 이용할 수 있다. 상기 티올 보호기는 메톡시메틸 (Methoxymethyl)기, 벤질옥시메틸 (Benzyloxymethyl)기, 트리페닐메틸 (Triphenylmethyl)기, 터트-부틸디메틸실릴 (tert-Butyldimethylsilyl)기, 트리페닐실릴 (Triphenylsily)기, 트리이소프로필실릴 (Triispropylsilyl)기, 파라-메톡시벤질 (para-Methylxybenzyl)기, 테트라히드로피란 (Tetrahydropyran)기, 테트라히드로퓨란 (Tetrahydrofuran)기, 터트-부틸 (tert-Butyl)기, 디페닐메틸 (Diphenylmethyl)기, 2-클로로트리틸 (2-Chlorotrityl)기, 벤질 (Benzyl)기, 4-메톡시벤질 (4-Methoxybenzyl)기, 알릴 (Allyl)기, 터트-부틸옥시카르보닐 (tert-Butyloxycarbonyl)기, 아세틸 (Acetyl)기, 벤조일 (Benzoyl)기 등을 예시할 수 있으며, 메톡시메틸 (Methoxymethyl)기, 벤질옥시메틸 (Benzyloxymethyl)기, 트리페닐메틸 (Triphenylmethyl)기, 터트-부틸디메틸실릴 (tert-Butyldimethylsilyl)기, 트리페닐실릴 (Triphenylsily)기 또는 트리이소프로필실릴 (Triisopropylsilyl)기를 이용하는 것이 바람직하며, 2-클로로트리틸 (2-Chlorotrityl)기 또는 트리페닐메틸 (Triphenylmethyl)기를 이용하는 것이 보다 바람직하며, 아미드 보호기가 트리페닐메틸 (Triphenylmethyl)기를 이용하는 것이 가장 바람직하지만 이에 국한되는 것은 아니다.In Formula Ib, R₃ may use a thiol protecting group commonly used in the art. The thiol protecting group is a methoxymethyl group, a benzyloxymethyl group, a triphenylmethyl group, a tert -butyldimethylsilyl group, a triphenylsilyl group, and a triiso group. Triispropylsilyl group, para-methoxybenzyl ( para -Methylxybenzyl) group, tetrahydropyran (Tetrahydropyran) group, tetrahydrofuran (Tetrahydrofuran) group, tert-butyl ( tert -Butyl) group, diphenylmethyl (Diphenylmethyl) ) group, 2-chlorotrityl (2-Chlorotrityl) group, benzyl (Benzyl) group, 4-methoxybenzyl (4-Methoxybenzyl) group, allyl (Allyl) group, tert -Butyloxycarbonyl group, acetyl (Acetyl) group, benzoyl (Benzoyl) group, etc. can be exemplified, and methoxymethyl (Methoxymethyl) group, benzyloxymethyl (Benzyloxymethyl) group, triphenylmethyl (Triphenylmethyl) group, tert-butyldimethylsilyl ( It is preferable to use a tert -Butyldimethylsilyl) group, a triphenylsily group, or a triisopropylsilyl group, and it is more preferable to use a 2-chlorotrityl (2-Chlorotrityl) group or a triphenylmethyl group. and it is most preferable to use a triphenylmethyl group as the amide protecting group, but is not limited thereto.
상기 화학식 I-b에서 R₄은 당 업계에서 통상적으로 이용하는 카르복실산의 수산기 보호기를 이용할 수 있다. 상기 티올 보호기는 메톡시메틸 (Methoxymethyl)기, 벤질옥시메틸 (Benzyloxymethyl)기, 트리페닐메틸 (Triphenylmethyl)기, 터트-부틸디메틸실릴 (tert-Butyldimethylsilyl)기, 트리페닐실릴 (Triphenylsily)기, 트리이소프로필실릴 (Triispropylsilyl)기, 파라-메톡시벤질 (para-Methylxybenzyl)기, 테트라히드로피란 (Tetrahydropyran)기, 테트라히드로퓨란 (Tetrahydrofuran)기, 터트-부틸 (tert-Butyl)기, 디페닐메틸 (Diphenylmethyl)기, 2-클로로트리틸 (2-Chlorotrityl)기, 벤질 (Benzyl)기, 4-메톡시벤질 (4-Methoxybenzyl)기, 알릴 (Allyl)기, 터트-부틸옥시카르보닐 (tert-Butyloxycarbonyl)기, 아세틸 (Acetyl)기, 벤조일 (Benzoyl)기 등을 예시할 수 있으며, 메톡시메틸 (Methoxymethyl)기, 벤질옥시메틸 (Benzyloxymethyl)기, 트리페닐메틸 (Triphenylmethyl)기, 터트-부틸디메틸실릴 (tert-Butyldimethylsilyl)기, 트리페닐실릴 (Triphenylsily)기 또는 트리이소프로필실릴 (Triisopropylsilyl)기를 이용하는 것이 바람직하며, 터트-부틸 (tert-Butyl) 또는 트리페닐메틸 (Triphenylmethyl)기를 이용하는 것이 보다 바람직하며, 티올 보호기가 터트-부틸 (tert-Butyl)기를 이용하는 것이 가장 바람직하지만 이에 국한되는 것은 아니다.In Formula Ib, R₄ may use a hydroxyl protecting group of a carboxylic acid commonly used in the art. The thiol protecting group is a methoxymethyl group, a benzyloxymethyl group, a triphenylmethyl group, a tert -butyldimethylsilyl group, a triphenylsilyl group, and a triiso group. Triispropylsilyl group, para-methoxybenzyl ( para -Methylxybenzyl) group, tetrahydropyran (Tetrahydropyran) group, tetrahydrofuran (Tetrahydrofuran) group, tert-butyl ( tert -Butyl) group, diphenylmethyl (Diphenylmethyl) ) group, 2-chlorotrityl (2-Chlorotrityl) group, benzyl (Benzyl) group, 4-methoxybenzyl (4-Methoxybenzyl) group, allyl (Allyl) group, tert -Butyloxycarbonyl group, acetyl (Acetyl) group, benzoyl (Benzoyl) group, etc. can be exemplified, and methoxymethyl (Methoxymethyl) group, benzyloxymethyl (Benzyloxymethyl) group, triphenylmethyl (Triphenylmethyl) group, tert-butyldimethylsilyl ( It is preferable to use a tert -Butyldimethylsilyl) group, a triphenylsily group, or a triisopropylsilyl group, and it is more preferable to use a tert-butyl ( tert -Butyl) or triphenylmethyl group, thiol It is most preferred that the protecting group uses a tert -Butyl group, but is not limited thereto.
이하, 상기 합성 방법을 단계별로 좀 더 상세히 설명한다.Hereinafter, the synthesis method will be described in more detail step by step.
고체상 펩타이드 합성방법은;Solid-phase peptide synthesis method;
1) 라이신이 결합된 레진을 Swelling하고 보호화된 두 번째 서열의 아미노산을 레진에 붙이는 단계;1) Swelling the lysine-bound resin and attaching the protected amino acid of the second sequence to the resin;
2) 아미노산 보호기(N-말단의 아미노기)를 염기(피페리딘)로 처리하여 제거하고 레진을 세척하는 단계;2) removing the amino acid protecting group (amino group at the N-terminus) by treatment with a base (piperidine) and washing the resin;
3) 결합을 위한 시약과 함께 보호화된 아미노산을 세척된 레진에 반응시켜 결합하는 단계;3) reacting the protected amino acid with a reagent for binding to the washed resin to bind;
4) 상기 2) 및 3)의 단계를 반복하여 서열 순서대로 합성하는 단계;4) repeating steps 2) and 3) to synthesize in sequence order;
5) 레진 및 구성 아미노산들의 보호기를 동시에 제거하는 단계; 5) simultaneously removing the protecting groups of the resin and constituent amino acids;
6) 펩타이드를 용매로 고체화하여 얻은 다음 정제하는 단계; 및6) a step of purifying the peptide obtained by solidifying it with a solvent; and
7) 정제된 펩타이드를 농축하고 동결건조하여 수득하는 단계를 포함할 수 있다. 7) concentrating the purified peptide and lyophilizing to obtain it.
단계 1)에 있어서, 사용할 수 있는 레진으로는 2-클로로트리틸클로라이드 레진 (2-Chlorotritylchloride Resin), 왕 레진 (Wang Resin), 디에이치피피 레진 (DHPP Resin, 4-(1',1'-dimethyl-1'-hydroxypropyl)phenoxyacetyl alanyl aminomethyl polystyrene), 피디디엠 레진 (PDDM Resin, Diphenyldiazomethane Resin), 사스린 레진 (SASRIN Resin, 2-Methoxy-4-alkoxybenzyl alcohol Resin)으로 구성된 군에서 선택되는 1 이상의 것을 들 수 있다. In step 1), the usable resins include 2-Chlorotritylchloride Resin, Wang Resin, and DHPP Resin (DHPP Resin, 4-(1',1'-) dimethyl-1'-hydroxypropyl)phenoxyacetyl alanyl aminomethyl polystyrene), PDDM resin (PDDM Resin, Diphenyldiazomethane Resin), and sathrin resin (SASRIN Resin, 2-Methoxy-4-alkoxybenzyl alcohol Resin) at least one selected from the group consisting of can be heard
단계 1)에 있어서, 사용되는 용매로는 디클로로메탄 (Dichloromethane), N,N-디메틸포름아마이드 (N,N-Dimethylformamide), N,N-디메틸아세트아마이드 (N,N-Dimethylacetamide), N-메틸피롤리돈 (N-Methylpyrrolidone), 클로로포름 (Chloroform), 1,2-디클로로에탄 (1,2-Dichloroethane), 테트라히드로퓨란 (Tetrahydrofurane), 1,4-디옥산 (1,4-Dioxane), 아세토니트릴 (Acetonitrile), 메탄올 (Methanol), 에탄올 (Ethanol), 이소프로판올 (Isopropanol), 에틸렌 글리콜 (Ethylene glycol), 메틸 아세테이트 (Methyl acetate), 에틸 아세테이트 (Ethyl acetate) 등으로 구성된 군에서 선택되는 1 이상의 것을 들 수 있다.In step 1), the solvent used is dichloromethane, N,N-dimethylformamide (N,N-Dimethylformamide), N,N-dimethylacetamide (N,N-Dimethylacetamide), N-methyl Pyrrolidone (N-Methylpyrrolidone), chloroform (Chloroform), 1,2-dichloroethane (1,2-Dichloroethane), tetrahydrofurane (Tetrahydrofurane), 1,4-dioxane (1,4-Dioxane), aceto At least one selected from the group consisting of nitrile (Acetonitrile), methanol (Methanol), ethanol (Ethanol), isopropanol (Isopropanol), ethylene glycol (Ethylene glycol), methyl acetate (Methyl acetate), ethyl acetate (Ethyl acetate), etc. can be heard
단계 2)에 있어서, 사용되는 보호기를 제거하는 염기로는 피페리딘 (Piperidine), 피롤리딘 (Pyrrolidine), 피페라진 (Piperazine), DBU (1,8-diazabicyclo[5.4.0]undec-7-ene), 4-메틸피페리딘 (4-Methylpiperidine), 몰포린 (Morpholine), 1-메틸-3-부틸이미다졸리윰 테트라플로오로보란 (1-methyl-3-butyl imidazolium BF4), 에탄올아민 (Ethanolamine), 시클로헥실아민 (Cyclohexylamine), 트리스(2-아미노에틸)아민 (Tris(2-aminoethyl)amine), 1,3-디시클로헥산비스-(메틸아민) (1,3-Dicyclohexanebis-(methylamine)), 1,4-비스-(3-아미노프로필)피페라진 (1,4-Bis-(3-aminopropyl)piperazine), 디에틸아민 (Diethylamine), 4-디메틸아미노피리딘 (4-Dimethylaminopyridine) 등의 유기 염기 및 수산화 리튬 (Lithium hydroxide), 수산화 나트륨 (Sodium Hydroxide), 수산화 칼슘 (Calcium Hydroxide), 수산화 칼륨 (Potassium Hydroxide) 등의 무기염기로 구성된 군에서 선택되는 1 이상의 것을 들 수 있다.In step 2), the base to remove the protecting group used is piperidine, pyrrolidine, piperazine, DBU (1,8-diazabicyclo[5.4.0]undec-7 -ene), 4-methylpiperidine, morpholine, 1-methyl-3-butylimidazolium tetrafluoroborane (1-methyl-3-butyl imidazolium BF4), ethanol Amine (Ethanolamine), cyclohexylamine (Cyclohexylamine), tris (2-aminoethyl) amine (Tris (2-aminoethyl) amine), 1,3-dicyclohexanebis- (methylamine) (1,3-Dicyclohexanebis- (methylamine)), 1,4-bis- (3-aminopropyl) piperazine (1,4-Bis- (3-aminopropyl) piperazine), diethylamine (Diethylamine), 4-dimethylaminopyridine (4-Dimethylaminopyridine) ) and organic bases such as lithium hydroxide, sodium hydroxide, calcium hydroxide, potassium hydroxide, and the like, and at least one selected from the group consisting of inorganic bases.
단계 3)에 있어서, 보호화된 아미노산이 결합할 때 사용하는 시약으로는 DCC (N,N-Dicyclohexylcarbodiimide), DIC (N,N-Diisopropylcarbodiimide), BOP (Benzotriazole-1-yl-oxy-tris-(dimethylamino)-phosphonium hexafluorophosphate), PyBOP (Benzotriazol-1-yl-oxytripyrrolidinophosphoniumhexafluorophosphate), PyBrOP (Bromo-tripyrrolidino-phosphonium hexafluorophosphate), PyAOP (7-Aza-benzotriazol-1-yloxy-tripyrrolidino-phosphonium hexafluorophosphate), PyOxim (Ethyl cyano(hydooxyimino)acetato-O2)-tri-(1-pyrrolidinyl)-phosphonium hexafluorophosphate), HBTU (O-Benzotriazole-N,N,N',N'-tetramethyluroniumhexafluorophosphate), HCTU (2-(6-Chloro-1H-benzotriazol-1-yl)-N,N,N',N',-tetramethylaminium hexafluorophosphate), HDMC (N-[(5-chloro-1H-benzotriazol-1-yl)-dimethylamino-morpholino]-uronium hexafluorophosphate N-oxide), TBTU (O-(Benzotriazol-1-yl)-N,N,N',N'-tetramethyluroniumtetrafluoroborate), HATU (2-(1H-7-Azabenzotriazol-1-yl)-1,1,3,3-tetramethyluroniumhexafluorophosphatemethanaminium), TATU (2-(1H-7-Azabenzotriazol-1-yl)-1,1,3,3-tetramethyluroniumtetrafluoroboratemethanaminium), CDI (carbonyldiimidazole), COMU (1-[1-(Cyano-2-ethoxy-2-oxoethylidene-aminooxy)-dimethylamino-morpholino]-uronium hexafluorophosphate), TOTT (2-(1-Oxy-pyridin-2-yl)-1,1,3,3-tetramethyl-isothiouronium tetrafluoroborate), EDCㅇHCl (N-(3-dimethylaminopropyl)-N`-ethylcarbodiimide hydrochloride), TFFH (Tetramethylfluoroformamidinium hexafluorophosphate), EEDQ (N-Ethoxycarbony-2-ethoxy-1,2-dihydro-quinoline), T3P (2-Propanephosphonic acid anhydride), DEPBT (3-(diethoxyphosphoryloxy)-1,2,3-benzotriazin-4(3H)-one), Oxyma (Ethyl cyanohydroxyiminoacetate), HOBt (1-Hydroxybenzotriazole), HOOBt(HODhbt, Hydroxy-3,4-dihydro-4-ox-1,2,3-benzo-triazine), BTC (bis-Trichloromethylcarbonate or Triphosgene), CDI (1,1'-Carbonyldiimidazole), 6-ClHOBt (1-Hydroxy-6-chloro-benzotriazole), HOAt (1-Hydroxyazabenzotriazole) HOSu (N-Hydroxysuccinimide) 등으로 구성된 군에서 선택되는 1 이상의 것을 들 수 있다.In step 3), the reagent used when the protected amino acid binds is DCC (N,N-Dicyclohexylcarbodiimide), DIC (N,N-Diisopropylcarbodiimide), BOP (Benzotriazole-1-yl-oxy-tris-( dimethylamino)-phosphonium hexafluorophosphate), PyBOP (Benzotriazol-1-yl-oxytripyrrolidinophosphoniumhexafluorophosphate), PyBrOP (Bromo-tripyrrolidino-phosphonium hexafluorophosphate), PyAOP (7-Aza-benzotriazol-1-yloxy-tripyrrolidino-phosphonium hexafluorophosphate), PyOxim (hydooxyimino)acetato-O2)-tri-(1-pyrrolidinyl)-phosphonium hexafluorophosphate), HBTU (O-Benzotriazole-N,N,N',N'-tetramethyluroniumhexafluorophosphate), HCTU (2-(6-Chloro-1H- benzotriazol-1-yl)-N,N,N',N',-tetramethylaminium hexafluorophosphate), HDMC (N-[(5-chloro-1H-benzotriazol-1-yl)-dimethylamino-morpholino]-uronium hexafluorophosphate N- oxide), TBTU (O-(Benzotriazol-1-yl)-N,N,N',N'-tetramethyluroniumtetrafluoroborate), HATU (2-(1H-7-Azabenzotriazol-1-yl)-1,1,3, 3-tetramethyluroniumhexafluorophosphatemethanaminium), TATU (2-(1H-7-Azabenzotriazol-1-yl)-1,1,3,3 -tetramethyluroniumtetrafluoroboratemethanaminium), CDI (carbonyldiimidazole), COMU (1-[1-(Cyano-2-ethoxy-2-oxoethylidene-aminooxy)-dimethylamino-morpholino]-uronium hexafluorophosphate), TOTT (2-(1-Oxy-pyridin- 2-yl)-1,1,3,3-tetramethyl-isothiouronium tetrafluoroborate), EDC∙HCl (N-(3-dimethylaminopropyl)-N`-ethylcarbodiimide hydrochloride), TFFH (Tetramethylfluoroformamidinium hexafluorophosphate), EEDQ (N-Ethoxycarbony- 2-ethoxy-1,2-dihydro-quinoline), T3P (2-Propanephosphonic acid anhydride), DEPBT (3-(diethoxyphosphoryloxy)-1,2,3-benzotriazin-4(3H)-one), Oxyma (Ethyl cyanohydroxyiminoacetate) ), HOBt (1-Hydroxybenzotriazole), HOOBt (HODhbt, Hydroxy-3,4-dihydro-4-ox-1,2,3-benzo-triazine), BTC (bis-Trichloromethylcarbonate or Triphosgene), CDI (1,1 and at least one selected from the group consisting of '-Carbonyldiimidazole), 6-ClHOBt (1-Hydroxy-6-chloro-benzotriazole), HOAt (1-Hydroxyazabenzotriazole) HOSu (N-Hydroxysuccinimide), and the like.
단계 5)에 있어서, 레진 및 아미노산의 보호기를 동시에 제거하는 과정은 산성 용액의 존재 하에서 수행한다. 상기 산성 용액은 TFA/phenol/water/TIPS (88/5/5/2), TFA/phenol/water/thioanisole/EDT (82.5/5/5/5/2.5), TFA/phenol/water/thioanisole/1-decanethiol (82.5/5/5/5/2.5), TFA/DTT/water/TIPS (88/5/5/2), TFA-DODT-TIS-anisole-water (183:5:2:5:5, v/v), TFA/phenol (95/5), TFA/phenol/Methanesulfonic acid (95/2.5/2.5), TFA/thioanisole/EDT/anisole (90/5/3/2), TFA/TES (95/5), TFA/water (95/5), TFA/DCM/indole (70/28/2), TFA/TIPS/water (95/2.5/2.5) 등의 용액을 예시할 수 있다.In step 5), the process of simultaneously removing the protecting groups of the resin and the amino acid is performed in the presence of an acidic solution. The acidic solution is TFA/phenol/water/TIPS (88/5/5/2), TFA/phenol/water/thioanisole/EDT (82.5/5/5/5/2.5), TFA/phenol/water/thioanisole/ 1-decanethiol (82.5/5/5/5/2.5), TFA/DTT/water/TIPS (88/5/5/2), TFA-DODT-TIS-anisole-water (183:5:2:5: 5, v/v), TFA/phenol (95/5), TFA/phenol/Methanesulfonic acid (95/2.5/2.5), TFA/thioanisole/EDT/anisole (90/5/3/2), TFA/TES (95/5), TFA/water (95/5), TFA/DCM/indole (70/28/2), and TFA/TIPS/water (95/2.5/2.5) solutions can be exemplified.
단계 6)에 있어서, Crude 펩타이드를 정제할 때 사용하는 용매로는 메탄올 (Methanol), 에탄올 (Ethanol), 이소프로판올 (Isopropanol), 아세토니트릴 (Acetonitrile) 및 정제수의 혼합용액이며, 이때 사용하는 산으로는 삼불화아세트산 (Trifluoroacetic acid), 아세트산 (Acetic acid), 포름산 (Formic acid)를 0.1% 내지 5% 이내로 사용하여 정제할 수 있다. In step 6), the solvent used when purifying the crude peptide is a mixed solution of methanol, ethanol, isopropanol, acetonitrile and purified water, and the acid used at this time is Trifluoroacetic acid, acetic acid, and formic acid may be used in an amount of 0.1% to 5%.
상기 작용기에 대한 보호기는 "Protecting Groups in Organic Synthesis (Greene and Wuts, John Wiley & Sons, 1991)"에 상세히 기재되어 있다.Protecting groups for these functional groups are described in detail in "Protecting Groups in Organic Synthesis (Greene and Wuts, John Wiley & Sons, 1991)".
본 명세서에서 용어 "펩타이드"는 펩타이드 결합에 의해 아미노산 잔기들이 서로 결합되어 형성된 선형의 분자를 의미한다.As used herein, the term “peptide” refers to a linear molecule formed by bonding amino acid residues to each other by peptide bonds.
본 발명에 따른 신규한 펩타이드는 강력한 항산화 활성(도 2)이 있고, 또한 우수한 콜라게나제 저해 활성(도 3)이 있었다.The novel peptide according to the present invention has strong antioxidant activity (FIG. 2) and also has excellent collagenase inhibitory activity (FIG. 3).
따라서, 또 하나의 양태로서, 본 발명은 상기 신규한 펩타이드를 포함하는 주름 개선 및 피부 노화 방지를 위한 조성물에 관한 것이다. 본 발명의 신규한 펩타이드를 유효성분으로 함유하는 주름 개선 및 피부 노화 방지용 조성물은 약학용, 화장용 또는 건강기능식품용으로 활용 가능하다.Therefore, as another aspect, the present invention relates to a composition for wrinkle improvement and skin aging prevention comprising the novel peptide. The composition for improving wrinkles and preventing skin aging containing the novel peptide of the present invention as an active ingredient can be used for pharmaceutical, cosmetic or health functional food.
상기 펩티드를 포함하는 약학용 조성물은, 각각 통상의 방법에 따라 산제, 과립제, 정제, 캡슐제, 현탁액, 에멀젼, 시럽, 에어로졸 등의 경구형 제형, 외용제, 좌제 및 멸균 주사용액의 형태로 제형화하여 사용될 수 있으나, 이에 제한되지 않는다.The pharmaceutical composition containing the peptide is formulated in the form of oral dosage forms such as powders, granules, tablets, capsules, suspensions, emulsions, syrups, aerosols, external preparations, suppositories, and sterile injection solutions, respectively, according to conventional methods. may be used, but is not limited thereto.
본 발명의 펩티드를 함유하는 약학용 조성물에 함유될 수 있는 담체, 부형제 및 희석제로는 락토오즈, 덱스트로즈, 수크로스, 덱스트린, 말토덱스트린, 솔비톨, 만니톨, 자일리톨, 에리스리톨, 말티톨, 전분, 아카시아 고무, 알지네이트, 젤라틴, 칼슘 포스페이트, 칼슘 실리케이트, 셀룰로즈, 메틸 셀룰로즈, 미정질 셀룰로스, 폴리비닐 피롤리돈, 물, 메틸히드록시벤조에이트, 프로필히드록시벤조에이트, 탈크, 마그네슘 스테아레이트 및 광물유를 들수 있으나, 이에 제한되지 않는다. 제제화할 경우에는 보통 사용하는 충진제, 증량제, 결합제, 습윤제, 붕해제, 계면활성제 등의 희석제 또는 부형제를 사용하여 조제되나, 이에 제한되지 않는다.Carriers, excipients and diluents that may be included in the pharmaceutical composition containing the peptide of the present invention include lactose, dextrose, sucrose, dextrin, maltodextrin, sorbitol, mannitol, xylitol, erythritol, maltitol, starch, acacia Gum, alginate, gelatin, calcium phosphate, calcium silicate, cellulose, methyl cellulose, microcrystalline cellulose, polyvinyl pyrrolidone, water, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate and mineral oil. However, the present invention is not limited thereto. In the case of formulation, it is prepared using diluents or excipients, such as commonly used fillers, extenders, binders, wetting agents, disintegrants, and surfactants, but is not limited thereto.
상기 펩티드를 포함하는 화장용 조성물은 피부, 두피 등에 사용할 수 있고, 피부 노화 방지, 주름 개선 효과를 위하여 사용할 수 있다.The cosmetic composition containing the peptide can be used for skin, scalp, etc., and can be used for skin aging prevention and anti-wrinkle effect.
본 발명의 화장용 조성물에 포함되는 펩타이드의 함량은 용도, 적용 형태, 사용 목적 및 소망하는 효과에 따라서 적절히 조절 가능하며, 함량 대비 효과를 고려하여, 예컨대 전체 조성물 중량에 대하여 0.0001 내지 99.9 중량% 이내에서 사용할 수 있다.The content of the peptide contained in the cosmetic composition of the present invention can be appropriately adjusted according to the use, application form, purpose of use and desired effect, and in consideration of the effect compared to the content, for example, within 0.0001 to 99.9% by weight based on the total weight of the composition can be used in
본 발명의 주름 개선, 피부 노화 방지를 위한 조성물은 비경구로 투여할 수 있으며, 경피에 도포에 의한 국부 투여 (Topical application) 방식으로 적용될 수 있다.The composition for wrinkle improvement and skin aging prevention of the present invention may be administered parenterally, and may be applied by topical application by percutaneous application.
상기 화장용 조성물은 피부 및 두피에 경피적으로 적용되고, 기초 화장품, 가슴 및 둔부 전용 크림, 메이크업 화장품, 바디 제품, 면도용 제품, 모발 제품 등을 포함한 모든 화장품 제품의 제조에 사용 가능한 조성물을 의미하는 것으로, 경구제, 스프레이, 현탁액, 유액, 크림, 젤, 폼 등의 형태로 제제화된 것일 수 있으나 그 형태에 특별한 제한이 없다.The cosmetic composition is applied percutaneously to the skin and scalp and refers to a composition that can be used in the manufacture of all cosmetic products including basic cosmetics, creams for chest and buttocks, makeup cosmetics, body products, shaving products, hair products, etc. It may be formulated in the form of oral preparations, sprays, suspensions, emulsions, creams, gels, foams, etc., but there is no particular limitation on the form.
또한, 상기 화장용 조성물은 유연 화장수, 수렴화장수, 영양화장수, 영양 크림, 마사지크림, 아이크림, 아이 에센스, 에센스, 클렌징크림, 클렌징로션, 클렌징폼, 클렌징 워터, 팩, 파우더, 보디로션, 보디 크림, 보디 에센스, 보디 세정제, 자외선 차단 크림, 염모제, 샴푸, 린스, 치약, 구강 청정제, 로션, 연고, 젤, 크림, 패치 및 분무제로 이루어진 군으로부터 선택되는 제형을 가지는 것을 포함한다.In addition, the cosmetic composition is a flexible lotion, astringent lotion, nourishing lotion, nourishing cream, massage cream, eye cream, eye essence, essence, cleansing cream, cleansing lotion, cleansing foam, cleansing water, pack, powder, body lotion, body creams, body essences, body washes, sunscreen creams, hair dyes, shampoos, rinses, toothpastes, mouthwashes, lotions, ointments, gels, creams, patches and sprays.
상기 펩타이드를 포함하는 기능성 식품 조성물은 정제, 캅셀, 분말, 과립, 액상, 환 등의 형태로 제조 및 가공할 수 있다. The functional food composition containing the peptide may be manufactured and processed in the form of tablets, capsules, powders, granules, liquids, pills, and the like.
본 발명의 건강기능식품이라 함은, 건강기능식품에 관한 법률 제6727호에 따른 인체에 유용한 기능성을 가진 원료나 성분을 사용하여 제조 및 가공한 식품을 말하며, 인체의 구조 및 기능에 대하여 영양소를 조절하거나 생리학적 작용 등과 같은 보건 용도에 유용한 효과를 얻을 목적으로 섭취하는 것을 의미한다.The health functional food of the present invention refers to food manufactured and processed using raw materials or ingredients useful for the human body according to Act No. 6727 of the Health Functional Food Act, and contains nutrients for the structure and function of the human body. It refers to ingestion for the purpose of obtaining useful effects for health purposes such as regulation or physiological action.
본 발명의 건강기능식품은 통상의 식품 첨가물을 포함할 수 있으며, 식품 첨가물로서의 적합 여부는 다른 규정이 없는 한, 식품의약품안전청에 승인된 식품 첨가물 공전의 총칙 및 일반시험법 등에 따라 해당 품목에 관한 규격 및 기준에 의하여 판정한다.The health functional food of the present invention may contain normal food additives, and unless otherwise specified, whether it is suitable as a food additive is related to the item according to the general rules and general test method of food additives approved by the Food and Drug Administration. It is judged according to the standards and standards.
상기 식품 첨가물 공전에 수재된 품목으로는 예를 들어, 케톤류, 글리신, 구연산칼슘, 니코틴산, 계피산 등의 화학적 합성물; 감색소, 감초추출물, 결정셀룰로오스, 고량색소, 구아검 등의 천연첨가물; L-글루타민산나트륨 제제, 면류첨가알칼리제, 보존료제제, 타르색소제제 등의 혼합제제류 등을 포함하나, 이에 제한되지 않는다.The items listed in the Food Additives Codex include, for example, chemical compounds such as ketones, glycine, calcium citrate, nicotinic acid, and cinnamic acid; natural additives such as persimmon pigment, licorice extract, crystalline cellulose, high pigment, and guar gum; Mixed preparations such as sodium L-glutamate preparations, noodle-added alkalis, preservatives, and tar dyes are included, but are not limited thereto.
또한, 상기 본 발명의 주름 개선 및 피부 노화 방지용 조성물은 상기 유효성분 이외에 통상의 제품화 및 제제화에 사용 가능한 모든 종류의 성분, 예컨대 향료, 색소, 살균제, 산화 방지제, 방부제, 보습제, 점증제, 부형제, 희석제, 무기염류 및 합성 고분자 물질 등을 추가로 포함할 수 있으며, 그 종류와 함량은 최종 산물의 용도 및 사용 목적에 따라 적절하게 조절할 수 있다.In addition, the composition for wrinkle improvement and skin aging of the present invention includes all kinds of ingredients that can be used for general commercialization and formulation in addition to the active ingredients, such as fragrances, pigments, bactericides, antioxidants, preservatives, moisturizers, thickeners, excipients, Diluents, inorganic salts, synthetic polymers, etc. may be additionally included, and the type and content may be appropriately adjusted according to the purpose and purpose of the final product.
본 발명의 신규한 펩타이드는 항산화 활성 및 콜라게나제 저해활성이 우수하여 주름 개선, 노화 방지 효과가 뛰어나므로, 인체 적용시 본래의 기능을 효과적으로 발휘할 수 있어, 의약, 화장품, 기능성 식품 분야에서 유용하게 이용될 수 있다.The novel peptide of the present invention has excellent antioxidant activity and collagenase inhibitory activity, so it has excellent anti-wrinkle and anti-aging effects. can be used
도 1은 본 발명의 실시 예에 따른 항산화 활성 및 콜라게나제 저해활성을 갖는 펩타이드를 합성하는 공정도이다.
도 2는 본 발명에 따른 신규한 펩타이드의 항산화 활성을 평가한 결과이다.
도 3은 본 발명에 따른 신규한 펩타이드의 콜라게나제 저해 활성을 평가한 결과이다. 1 is a flowchart of synthesizing a peptide having antioxidant activity and collagenase inhibitory activity according to an embodiment of the present invention.
2 is a result of evaluating the antioxidant activity of the novel peptide according to the present invention.
3 is a result of evaluating the collagenase inhibitory activity of the novel peptide according to the present invention.
본 명세서에서 특별한 표시가 없는 한, 아미노산 및 보호기의 지정에 사용되는 약어는 IUPAC-IUB의 생화학 용어 위원회 (Commission of Biochemical Nomenclature)에서 권장하는 용어에 기초한다 (Biochemistry, 11:1726-1732(1972); Pure & Appl. Chem., Vol. 56, No. 5, pp. 595-624, 1984).Unless otherwise indicated herein, abbreviations used for designation of amino acids and protecting groups are based on terms recommended by the IUPAC-IUB Commission of Biochemical Nomenclature ( Biochemistry, 11:1726-1732 (1972)) ( Pure & Appl. Chem., Vol. 56, No. 5, pp. 595-624, 1984).
본 명세서에서 사용한 보호기 및 아미노산의 약어는 다음과 같다:Abbreviations of protecting groups and amino acids used herein are as follows:
AcOH: 아세틱 엑시드 (Acetic acid)AcOH: Acetic acid
Cys: 시스테인 (Cysteine)Cys: Cysteine
DCM: 디클로로메탄 (Dichloromethane)DCM: Dichloromethane
DMSO: 디메틸설폭시드 (Dimethylsulfoxide)DMSO: Dimethylsulfoxide
DTT: 디티올쓰레이톨 (Dithiolthreitol)DTT: Dithiolthreitol
EDT: 1,2-에탄디티올 (1,2-Ethanedithiol)EDT: 1,2-ethanedithiol (1,2-Ethanedithiol)
Gly: 글라이신 (Glycine)Gly: Glycine
γ-Glu: 감마-글루타믹 산 (γ-Glutamic acid)γ-Glu: gamma-glutamic acid (γ-Glutamic acid)
His: 히스티딘 (Histidine)His: Histidine
HPLC: 고성능액체크로마토그래피 (High Performance Liquid Chromatography)HPLC: High Performance Liquid Chromatography
Lys: 라이신 (Lysine)Lys: Lysine
Boc: 터트-부틸옥시카르보닐기 (tert-Butyloxycarbonyl)Boc: tert-butyloxycarbonyl group ( tert -Butyloxycarbonyl)
OtBu: O-터트-부틸(O-tert-Butyl)OtBu: O- tert -Butyl
t-Bu: 터트-부틸 (tert-Butyl)t-Bu: tert -Butyl
Fmoc: 9-플루오레닐옥시카보닐 (9-Fluorenyloxycarbonyl)Fmoc: 9-fluorenyloxycarbonyl (9-Fluorenyloxycarbonyl)
Trt: 트리페닐메틸 (또는 트리틸) (Triphenylmethyl or Trityl)Trt: triphenylmethyl (or trityl) (Triphenylmethyl or Trityl)
Pbf: 2,2,4,6,7-펜타메틸-디히드로벤조퓨란-5-설포닐 (2,2,4,6,7-Pentamethyl-dihydrobenzofuran-5-sulfonyl)Pbf: 2,2,4,6,7-pentamethyl-dihydrobenzofuran-5-sulfonyl (2,2,4,6,7-Pentamethyl-dihydrobenzofuran-5-sulfonyl)
Pmc: 2,2,5,7,8-펜타메틸크로만-6-설포닐 (2,2,5,7,8-Pentamethylchroman-6-sulfonyl)Pmc: 2,2,5,7,8-pentamethylchroman-6-sulfonyl (2,2,5,7,8-Pentamethylchroman-6-sulfonyl)
Mtr: 4-메톡시-2,3,6-트리메틸페닐-설포닐 (4-Methoxy-2,3,6-trimethylphenyl-sulfonyl)Mtr: 4-Methoxy-2,3,6-trimethylphenyl-sulfonyl (4-Methoxy-2,3,6-trimethylphenyl-sulfonyl)
Tos: 파라-톨루엔설포닐 (para-Toluenesulfonyl)Tos: para-toluenesulfonyl (para-Toluenesulfonyl)
TES: 트라이에틸실란 (Triethylsilane)TES: Triethylsilane
TIS: 트리이소프로필실란 (Triisopropylsilane)TIS: Triisopropylsilane
TFA: 트라이플루오로아세틱 엑시드 (Trifluoroacetic acid)TFA: Trifluoroacetic acid
TIPS: 트리이소프로필실란 (Triisopropylsilane)TIPS: Triisopropylsilane
이하, 본 발명을 실시 예에 의해 더욱 상세히 설명하고자 한다. 이들 실시 예는 오로지 본 발명을 보다 구체적으로 설명하기 위한 것으로, 본 발명의 요지에 따라 본 발명의 범위가 이들 실시 예에 의해 제한되지 않는다는 것은 당 업계에서 통상의 지식을 가진 자에 있어서 자명할 것이다.Hereinafter, the present invention will be described in more detail by way of examples. These examples are only for illustrating the present invention in more detail, and it will be apparent to those of ordinary skill in the art that the scope of the present invention is not limited by these examples according to the gist of the present invention. .
실시 예. 화학식 I로 표시되는 펩타이드의 제조Example. Preparation of the peptide represented by the formula (I)
본 명세서 전체에 거쳐, 특정 물질의 농도를 나타내기 위하여 사용되는 "%"는 별도의 언급이 없는 한 고체/고체는 (중량/중량) %, 고체/액체는 (중량/부피) %, 그리고 액체/액체는 (부피/부피) %이다.Throughout this specification, "%" used to indicate the concentration of a specific substance is (weight/weight) % for solids/solids, (weight/volume) % for solids/liquids, and liquids unless otherwise specified. /liquid is (volume/volume) %.
실시 예 1 : H-His(Trt)-Lys(Boc)-Wang Resin의 제조Example 1: Preparation of H-His(Trt)-Lys(Boc)-Wang Resin
[화학식 2][Formula 2]
H-Lys(Boc)-Wang 레진 200g (치환율=0.5, 100 mmole)이 들어있는 반응기에 Fmoc-His(Trt)-OH (분자량 = 619.7 g/mol, 185.91 g, 300 mmole)과 Oxyma (Ethyl cyanohydroxyiminoacetate, 분자량 = 142.11 g/mol, 46.9 g, 330 mmole)을 THF 1000 ml에 용해한 다음 투입하였다. 혼합된 반응액에 DIC (분자량 = 126.2 g/mol, 41.7 g, 330 mmole)을 서서히 적가한 다음 상온에서 4시간 동안 서서히 교반하였다. 레진을 THF 1000 ml으로 2회 세척한 후, 20% 피페리딘/THF 용액 1000 ml를 더하고 15분 동안 교반하였다. 감압 여과하여 반응액을 제거하고, 레진을 동일하게 1회 더 처리한다. THF 1000 ml으로 총 6회 레진을 세척한 후, 화학식 2로 표시되는 H-His(Trt)-Lys(Boc)-Wang 레진을 수득하였다 (수율 >99%).In a reactor containing 200 g of H-Lys(Boc)-Wang resin (substitution ratio = 0.5, 100 mmole), Fmoc-His(Trt)-OH (molecular weight = 619.7 g/mol, 185.91 g, 300 mmole) and Oxyma (Ethyl cyanohydroxyiminoacetate) , molecular weight = 142.11 g/mol, 46.9 g, 330 mmole) was dissolved in 1000 ml of THF and then added. DIC (molecular weight = 126.2 g/mol, 41.7 g, 330 mmole) was slowly added dropwise to the mixed reaction solution, followed by stirring slowly at room temperature for 4 hours. After the resin was washed twice with 1000 ml of THF, 1000 ml of a 20% piperidine/THF solution was added and stirred for 15 minutes. The reaction solution is removed by filtration under reduced pressure, and the resin is treated in the same way once more. After washing the resin a total of 6 times with 1000 ml of THF, H-His(Trt)-Lys(Boc)-Wang resin represented by
실시 예 2 : H-Gly-His(Trt)-Lys(Boc)-Wang Resin의 제조Example 2: Preparation of H-Gly-His(Trt)-Lys(Boc)-Wang Resin
[화학식 3][Formula 3]
H-His(Trt)-Lys(Boc)-Wang 레진 (100 mmole)이 들어있는 반응기에 Fmoc-Gly-OH (분자량 = 297.3 g/mol, 89.19 g, 300 mmole)과 Oxyma (Ethyl cyanohydroxyiminoacetate, 분자량 = 142.11 g/mol, 46.9 g, 330 mmole)을 THF 1000 ml에 용해한 다음 투입하였다. 혼합된 반응액에 DIC (분자량 = 126.2 g/mol, 41.7 g, 330 mmole)을 적가한 다음 상온에서 5시간 동안 서서히 교반하였다. 레진을 THF 1000 ml으로 2회 세척한 후, 20% 피페리딘/THF 용액 1000 ml를 더하고 15분 동안 교반한다. 감압 여과하여 반응액을 제거하고, 레진을 동일하게 1회 더 처리하였다. THF 1000 ml으로 총 6회 레진을 세척한 후 화학식 3으로 표시되는 H-Gly-His(Trt)-Lys(Boc)-Wang 레진을 수득하였다 (수율 >99%).In a reactor containing H-His(Trt)-Lys(Boc)-Wang resin (100 mmole), Fmoc-Gly-OH (molecular weight = 297.3 g/mol, 89.19 g, 300 mmole) and Oxyma (Ethyl cyanohydroxyiminoacetate, molecular weight = 142.11 g/mol, 46.9 g, 330 mmole) was dissolved in 1000 ml of THF and then added. DIC (molecular weight = 126.2 g/mol, 41.7 g, 330 mmole) was added dropwise to the mixed reaction solution, followed by slow stirring at room temperature for 5 hours. After washing the resin twice with 1000 ml of THF, 1000 ml of a 20% piperidine/THF solution is added and stirred for 15 minutes. The reaction solution was removed by filtration under reduced pressure, and the resin was treated in the same way once more. After washing the resin a total of 6 times with 1000 ml of THF, H-Gly-His(Trt)-Lys(Boc)-Wang resin represented by
실시 예 3 : H-Cys(Trt)-Gly-His(Trt)-Lys(Boc)-Wang Resin의 제조Example 3: Preparation of H-Cys(Trt)-Gly-His(Trt)-Lys(Boc)-Wang Resin
[화학식 4][Formula 4]
H-Gly-His(Trt)-Lys(Boc)-Wang 레진 (100 mmole)이 들어있는 반응기에 Fmoc-Cys(Trt)-OH (분자량 = 585.7 g/mol, 175.71 g, 300 mmole)과 Oxyma (Ethyl cyanohydroxyiminoacetate, 분자량 = 142.11 g/mol, 46.9 g, 330 mmole)을 THF 1000 ml에 용해한 다음 투입하였다. 혼합된 반응액에 DIC (분자량 = 126.2 g/mol, 41.7 g, 330 mmole)을 적가한 다음 상온에서 5시간 동안 서서히 교반하였다. 레진을 THF 1000 ml으로 2회 세척한 후, 20% 피페리딘/THF 용액 1000 ml를 더하고 15분 동안 교반하였다. 감압 여과하여 반응액을 제거하고, 레진을 동일하게 1회 더 처리하였다. THF 1000 ml으로 총 6회 레진을 세척한 후 화학식 4로 표시되는 H-Cys(Trt)-Gly-His(Trt)-Lys(Boc)-Wang 레진을 수득하였다 (수율 >99%).Fmoc-Cys(Trt)-OH (molecular weight = 585.7 g/mol, 175.71 g, 300 mmole) and Oxyma ( Ethyl cyanohydroxyiminoacetate, molecular weight = 142.11 g/mol, 46.9 g, 330 mmole) was dissolved in 1000 ml of THF and then added. DIC (molecular weight = 126.2 g/mol, 41.7 g, 330 mmole) was added dropwise to the mixed reaction solution, followed by slow stirring at room temperature for 5 hours. After the resin was washed twice with 1000 ml of THF, 1000 ml of a 20% piperidine/THF solution was added and stirred for 15 minutes. The reaction solution was removed by filtration under reduced pressure, and the resin was treated in the same way once more. After washing the resin a total of 6 times with 1000 ml of THF, H-Cys(Trt)-Gly-His(Trt)-Lys(Boc)-Wang resin represented by Chemical Formula 4 was obtained (yield >99%).
실시 예 4 : H-γ-Glu-(OtBu)-Cys(Trt)-Gly-His(Trt)-Lys(Boc)-Wang Resin 의 제조Example 4: Preparation of H-γ-Glu-(OtBu)-Cys(Trt)-Gly-His(Trt)-Lys(Boc)-Wang Resin
[화학식 5][Formula 5]
H-Cys(Trt)-Gly-His(Trt)-Lys(Boc)-Wang 레진 (100 mmole)이 들어있는 반응기에 Fmoc-γ-Glu-(OtBu) (분자량 = 425.5 g/mol, 127.65 g, 300 mmole)과 Oxyma (Ethyl cyanohydroxyiminoacetate, 분자량 = 142.11 g/mol, 46.9 g, 330 mmole)을 DMF 1000 ml에 용해한 다음 투입하였다. 혼합된 반응액에 DIC (분자량 = 126.2 g/mol, 41.7 g, 330 mmole)을 적가한 다음 상온에서 5시간 동안 서서히 교반하였다. 레진을 THF 1000 ml으로 2회 세척한 후, 20% 피페리딘/THF 용액 1000 ml를 더하고 15분 동안 교반하였다. 감압 여과하여 반응액을 제거하고, 레진을 동일하게 1회 더 처리하였다. THF 1000 ml으로 총 6회 레진을 세척한 후 화학식 5로 표시되는 H-γ-Glu-(OtBu)-Cys(Trt)-Gly-His(Trt)-Lys(Boc)-Wang 레진을 수득하였다 (수율 >99%).Fmoc-γ-Glu-(OtBu) (molecular weight = 425.5 g/mol, 127.65 g, 300 mmole) and Oxyma (Ethyl cyanohydroxyiminoacetate, molecular weight = 142.11 g/mol, 46.9 g, 330 mmole) were dissolved in 1000 ml of DMF and then added. DIC (molecular weight = 126.2 g/mol, 41.7 g, 330 mmole) was added dropwise to the mixed reaction solution, followed by slow stirring at room temperature for 5 hours. After the resin was washed twice with 1000 ml of THF, 1000 ml of a 20% piperidine/THF solution was added and stirred for 15 minutes. The reaction solution was removed by filtration under reduced pressure, and the resin was treated in the same way once more. After washing the resin a total of 6 times with 1000 ml of THF, H-γ-Glu-(OtBu)-Cys(Trt)-Gly-His(Trt)-Lys(Boc)-Wang resin represented by
실시 예 5 : H-Cys(Trt)-γ-Glu-(OtBu)-Cys(Trt)-Gly-His(Trt)-Lys(Boc)-Wang Resin의 제조Example 5: Preparation of H-Cys(Trt)-γ-Glu-(OtBu)-Cys(Trt)-Gly-His(Trt)-Lys(Boc)-Wang Resin
[화학식 6][Formula 6]
H-γ-Glu-(OtBu)-Cys(Trt)-Gly-His(Trt)-Lys(Boc)-Wang 레진 (100 mmole)이 들어있는 반응기에 Fmoc-Cys(Trt)-OH (분자량 = 585.7 g/mol, 175.71 g, 300 mmole)과 Oxyma (Ethyl cyanohydroxyiminoacetate, 분자량 = 142.11 g/mol, 46.9 g, 330 mmole)을 THF 1000 ml에 용해한 다음 투입하였다. 혼합된 반응액에 DIC (분자량 = 126.2 g/mol, 41.7 g, 330 mmole)을 적가한 다음 상온에서 4시간 동안 서서히 교반하였다. 레진을 THF 1000 ml으로 2회 세척한 후, 20% 피페리딘/THF 용액 1000 ml를 더하고 15분 동안 교반하였다. 감압 여과하여 반응액을 제거하고, 레진을 동일하게 1회 더 처리하였다. THF 1000 ml으로 총 6회 레진을 세척한 후 화학식 6으로 표시되는 H-Cys(Trt)-γ-Glu-(OtBu)-Cys(Trt)-Gly-His(Trt)-Lys(Boc)-Wang 레진을 수득하였다 (수율 >99%).In a reactor containing H-γ-Glu-(OtBu)-Cys(Trt)-Gly-His(Trt)-Lys(Boc)-Wang resin (100 mmole) Fmoc-Cys(Trt)-OH (molecular weight = 585.7 g/mol, 175.71 g, 300 mmole) and Oxyma (Ethyl cyanohydroxyiminoacetate, molecular weight = 142.11 g/mol, 46.9 g, 330 mmole) were dissolved in 1000 ml of THF and then added. DIC (molecular weight = 126.2 g/mol, 41.7 g, 330 mmole) was added dropwise to the mixed reaction solution, followed by slow stirring at room temperature for 4 hours. After the resin was washed twice with 1000 ml of THF, 1000 ml of a 20% piperidine/THF solution was added and stirred for 15 minutes. The reaction solution was removed by filtration under reduced pressure, and the resin was treated in the same way once more. After washing the resin a total of 6 times with 1000 ml of THF, H-Cys(Trt)-γ-Glu-(OtBu)-Cys(Trt)-Gly-His(Trt)-Lys(Boc)-Wang represented by Formula 6 A resin was obtained (yield >99%).
실시 예 6 : H-γ-Glu-(OtBu)-Cys(Trt)-γ-Glu-(OtBu)-Cys(Trt)-Gly-His(Trt) -Lys(Boc)-Wang Resin의 제조Example 6: Preparation of H-γ-Glu-(OtBu)-Cys(Trt)-γ-Glu-(OtBu)-Cys(Trt)-Gly-His(Trt)-Lys(Boc)-Wang Resin
[화학식 7][Formula 7]
H-Cys(Trt)-γ-Glu-(OtBu)-Cys(Trt)-Gly-His(Trt)-Lys(Boc)-Wang 레진 (100 mmole)이 들어있는 반응기에 Fmoc-γ-Glu-(OtBu) (분자량 = 425.5 g/mol, 127.65 g, 300 mmole)과 Oxyma (Ethyl cyanohydroxyiminoacetate, 분자량 = 142.11 g/mol, 46.9 g, 330 mmole)을 THF 1000 ml에 용해한 다음 투입하였다. 혼합된 반응액에 DIC (분자량 = 126.2 g/mol, 41.7 g, 330 mmole)을 적가한 다음 상온에서 5시간 동안 서서히 교반하였다. 레진을 THF 1000 ml으로 2회 세척한 후, 20% 피페리딘/THF 용액 1000 ml를 더하고 15분 동안 교반하였다. 감압 여과하여 반응액을 제거하고, 레진을 동일하게 1회 더 처리하였다. THF 1000 ml으로 총 6회 레진을 세척한 후 화학식 7로 표시되는 H-γ-Glu-(OtBu)-Cys(Trt)-γ-Glu-(OtBu)-Cys(Trt)-Gly-His(Trt)-Lys(Boc)-Wang 레진을 수득하였다 (수율 >99%).Fmoc-γ-Glu-( OtBu) (molecular weight = 425.5 g/mol, 127.65 g, 300 mmole) and Oxyma (Ethyl cyanohydroxyiminoacetate, molecular weight = 142.11 g/mol, 46.9 g, 330 mmole) were dissolved in 1000 ml of THF and then added. DIC (molecular weight = 126.2 g/mol, 41.7 g, 330 mmole) was added dropwise to the mixed reaction solution, followed by slow stirring at room temperature for 5 hours. After the resin was washed twice with 1000 ml of THF, 1000 ml of a 20% piperidine/THF solution was added and stirred for 15 minutes. The reaction solution was removed by filtration under reduced pressure, and the resin was treated in the same way once more. After washing the resin a total of 6 times with 1000 ml of THF, H-γ-Glu-(OtBu)-Cys(Trt)-γ-Glu-(OtBu)-Cys(Trt)-Gly-His(Trt )-Lys(Boc)-Wang resin was obtained (yield >99%).
실시 예 7 : Crude H-γ-Glu-Cys-γ-Glu-Cys-Gly-His-Lys-OH의 제조Example 7: Preparation of Crude H-γ-Glu-Cys-γ-Glu-Cys-Gly-His-Lys-OH
H-γ-Glu-(OtBu)-Cys(Trt)-γ-Glu-(OtBu)-Cys(Trt)-Gly-His(Trt)-Lys(Boc)-Wang 레진을 반응기에 투입하고 냉각된 TFA/EDT/TIS/water (92.5/2.5/2.5/2.5) 용액 3 L를 서서히 부어 넣은 다음 상온에서 4시간 동안 교반하였다. 반응액을 받아낸 다음 냉각된 디에틸에테르 12 L에 반응액을 서서히 적가하여 펩타이드를 석출시켰다. 실온에서 30분 동안 교반한 다음 여과하여 펩타이드를 회수하였다. 최대한 용매를 감압 제거한 다음 진공 건조기에서 5시간 동안 건조 시켰다. 거의 백색의 Crude peptide 83.7 g을 수득하였다. (분자량 804.88g/mol, Crude peptide 순도 72.6%, 수율 104.0%)H-γ-Glu-(OtBu)-Cys(Trt)-γ-Glu-(OtBu)-Cys(Trt)-Gly-His(Trt)-Lys(Boc)-Wang resin was put into the reactor and cooled TFA 3 L of /EDT/TIS/water (92.5/2.5/2.5/2.5) solution was slowly poured thereinto, followed by stirring at room temperature for 4 hours. After receiving the reaction solution, the reaction solution was slowly added dropwise to 12 L of cooled diethyl ether to precipitate the peptide. After stirring at room temperature for 30 minutes, the peptide was recovered by filtration. After removing the solvent under reduced pressure as much as possible, it was dried in a vacuum dryer for 5 hours. 83.7 g of almost white crude peptide was obtained. (Molecular weight 804.88g/mol, crude peptide purity 72.6%, yield 104.0%)
실시 예 8 : H-γ-Glu-Cys-γ-Glu-Cys-Gly-His-Lys-OH의 제조Example 8: Preparation of H-γ-Glu-Cys-γ-Glu-Cys-Gly-His-Lys-OH
[화학식 I][Formula I]
상기 실시 예 7에서 수득한 Crude 펩타이드 83.7 g을 정제수에 용해한 다음 0.46μm membrane 여과하였다. 여과액을 산업용 HPLC (230 nm, 500 ml/분, 10 미크론 C18 컬럼에서 20분 내에 0.1% TFA 내 아세토니트릴 초기농도 8%에서 55%로 증가)로 반복 주입한 후 분리 정제하여 화학식 I로 표시되는 펩타이드, 42.3 g (분자량 804.88g/mol, 수율: 50.5%, 순도: 97.5%)을 수득하였다.83.7 g of the crude peptide obtained in Example 7 was dissolved in purified water and then filtered with a 0.46 μm membrane. The filtrate was repeatedly injected with industrial HPLC (230 nm, 500 ml/min, increased from 8% to 55% initial concentration of acetonitrile in 0.1% TFA in 20 minutes on a 10 micron C18 column from 8% to 55%), followed by separation and purification, and represented by Formula I The peptide, 42.3 g (molecular weight 804.88 g/mol, yield: 50.5%, purity: 97.5%) was obtained.
실험 예 1: 신규 펩타이드 항산화능 평가 (ABTS 라디칼 소거 활성 평가)Experimental Example 1: Evaluation of antioxidant activity of novel peptides (ABTS radical scavenging activity evaluation)
실시예 8에서 수득한 펩타이드 소재의 항산화 활성평가를 위하여 ABTS (2,2′-Azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) diammonium salt)를 Potassium persulfate와 반응시켜 ABTS를 라디칼 상태로 산화시켜, 짙은 청색을 형성시켰다. In order to evaluate the antioxidant activity of the peptide material obtained in Example 8, ABTS (2,2′-Azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) diammonium salt) was reacted with potassium persulfate to oxidize ABTS to a radical state. , forming a dark blue color.
이후 항산화 소재와의 반응시키면 색을 잃어 투명해지는 원리를 이용하여 well plate reader 상에서 흡광도를 측정하여 항산화력을 측정하였다. 우선 ABTS와 potassium persulfate를 증류수에 녹여 각각 14 mM과 4.9 mM의 농도로 준비하고 1:1로 혼합하고 알루미늄 호일로 빛을 차단한 상태로 상온에서 12시간 동안 반응시켰다. After that, the absorbance was measured on a well plate reader using the principle of losing color and becoming transparent when reacted with an antioxidant material to measure antioxidant power. First, ABTS and potassium persulfate were dissolved in distilled water to prepare concentrations of 14 mM and 4.9 mM, respectively, mixed 1:1, and reacted for 12 hours at room temperature while blocking light with aluminum foil.
해당 용액을 1X PBS로 약 50배 희석하여 흡광광도계로 734 nm에서의 흡광도를 측정했을 때 값이 0.7(ㅁ0.02)이 되도록 하여 ABTS 라디칼 용액을 준비하고, 96 well plate에 여러 농도의 시료 10 μL와 ABTS 라디칼 용액 190μL을 넣고 알루미늄 호일로 차광하여 상온에서 5분간 방치한 후 734 nm에서의 흡광도를 측정하고, 그 결과를 도 2에 나타내었다.Prepare the ABTS radical solution by diluting the solution approximately 50 times with 1X PBS so that the value becomes 0.7 (w0.02) when the absorbance at 734 nm is measured with an absorbance meter, and 10 μL of samples of various concentrations in a 96 well plate and 190 μL of ABTS radical solution, shielded with aluminum foil, left at room temperature for 5 minutes, and then absorbance at 734 nm was measured, and the results are shown in FIG. 2 .
도 2에 도시된 바와 같이, 본 발명에 따른 헵타펩타이드는 100μM 이상의 농도에서 항산화 활성이 우수한 것을 알 수 있었다.As shown in FIG. 2 , it was found that the heptapeptide according to the present invention had excellent antioxidant activity at a concentration of 100 μM or more.
실험 예 2: 세포내 콜라게나제 활성 억제시험(Collagenase inhibition assay)Experimental Example 2: Intracellular collagenase activity inhibition assay (Collagenase inhibition assay)
실시예 8에서 수득한 펩타이드 소재에 의한 Procollagen 단백질 발현량 분석은 ELISA (enzyme-linked immunosorbent assay) 기반의 Procollagen Type I C-peptide (PIP) EIA Kit (Takara)을 이용하여 수행하였다. Procollagen protein expression level analysis by the peptide material obtained in Example 8 was performed using an ELISA (enzyme-linked immunosorbent assay)-based Procollagen Type I C-peptide (PIP) EIA Kit (Takara).
인간유래 피부각질형성세포주인 HaCaT cell을 6 well plate에 2×105개씩 분주하여 배양하고, 24시간 후 배지를 제거하고 무혈청 배지에서 시료를 처리하여 24시간 동안 배양하였다. 이후 배양 배지를 원심 분리하여 상등액만 얻어내고 이를 ELISA well-plate에 처리하여 450 nm에서의 흡광도를 측정해 Procollagen을 정량하여, 시료가 처리되지 않은 상태에서 발현된 procollagen의 양을 비교 대조군으로 설정하여 백분율로 나타내었다.HaCaT cells, a human-derived keratinocyte cell line, were aliquoted into 6 well plates by 2×10 5 and cultured. After 24 hours, the medium was removed and the sample was treated in a serum-free medium and cultured for 24 hours. Then, the culture medium was centrifuged to obtain only the supernatant, treated with an ELISA well-plate, and the absorbance at 450 nm was measured to quantify procollagen. The amount of procollagen expressed in the untreated state was set as a comparative control. expressed as a percentage.
이 시험방법은 섬유아세포(fibroblast) 배양시 시료가 세포내 콜라게나제 생성억제 정도를 공시료액과 비교하는 것으로서, 세부 방법은 다음과 같다.This test method compares the degree of inhibition of intracellular collagenase production of a sample during fibroblast culture with a blank sample solution. The detailed method is as follows.
(가) 시험방법(A) Test method
섬유아세포를 48-well plate에 well당 5×104 개로 분주한 다음 세포배양조건에서 24시간 배양 후, 배지를 버리고 10% 인산완충식염액(phosphate buffered saline(PBS))로 세척한 다음 검액 및 새로운 배지를 넣고 48시간 배양하였다. 배양액을 취하여 콜라게나제 양을 측정하고, 측정된 콜라게나제 양은 로우리법으로 구한 총 단백질 양으로 보정하였다. Fibroblasts were aliquoted into a 48-well plate at 5×10 4 cells per well, and after 24 hours of incubation in cell culture conditions, the medium was discarded, washed with 10% phosphate buffered saline (PBS), and the sample solution and A fresh medium was added and cultured for 48 hours. The culture medium was taken and the amount of collagenase was measured, and the measured amount of collagenase was corrected with the total protein amount obtained by the Lowry method.
(나) 콜라게나제양 측정(B) Measurement of collagenase amount
콜라게나제양은 하기의 ELISA법으로 측정하였다.The amount of collagenase was measured by the following ELISA method.
정량용 완충액2 100㎕를 well에 넣은 다음 1/5로 희석한 배양액 및 표준액 각 100㎕를 넣고 상온에서 2시간 배양한다. well에서 배양액을 제거한 다음 세척용 완충액 400㎕로 3회 워싱(washing)하였다. antiserum 100㎕를 넣고 상온에서 2시간 반응시킨 다음 세척용 완충액 400㎕로 3회 워싱(washing)하였다. Peroxidase conjugate solution 100㎕를 넣고 상온에서 2시간 반응시킨 다음 인산염완충액(PBS) 400㎕로 3회 워싱(washing)하였다. TMB substrate 100㎕를 넣고 상온에서 30분간 반응시킨 후, 1M 황산 100㎕를 넣고 450nm에서 ELISA reader로 측정하고 그 결과를 도 3에 나타내었다.100 μl of
도 3에 도시된 바와 같이, 본 발명에 따른 헵타펩타이드는 Adenosine과 비교하여 콜라게나제 저해활성이 우수한 것을 알 수 있었다.As shown in FIG. 3 , it was found that the heptapeptide according to the present invention had superior collagenase inhibitory activity compared to adenosine.
이상으로 본 발명 내용의 특정한 부분을 상세히 기술하였는 바, 당 업계의 통상의 지식을 가진 자에게 있어서 이러한 구체적 기술은 단지 바람직한 실시 양태일 뿐이며, 이에 의해 본 발명의 범위가 제되는 것이 아닌 점은 명백할 것이다. 따라서, 본 발명의 실질적인 범위는 첨부된 청구항들과 그것들의 등가물에 의하여 정의된다고 할 것이다.As the specific parts of the present invention have been described in detail above, for those of ordinary skill in the art, it is clear that these specific descriptions are only preferred embodiments, and the scope of the present invention is not limited thereby. will be. Accordingly, it is intended that the substantial scope of the present invention be defined by the appended claims and their equivalents.
Claims (5)
[화학식 I]
a peptide having the structure of formula (I);
[Formula I]
The peptide according to claim 1, wherein the peptide has antioxidant activity and collagenase inhibitory activity.
A composition for improving wrinkles and preventing skin aging comprising the peptide of claim 1.
The composition for improving wrinkles and preventing skin aging according to claim 3, wherein the composition has antioxidant activity and collagenase inhibitory activity.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| KR1020200162593A KR102506021B1 (en) | 2020-11-27 | 2020-11-27 | The Peptide having Excellent Antioxidant Activity and Collagenase Inhibition Activity and Composition Containing the same |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| KR1020200162593A KR102506021B1 (en) | 2020-11-27 | 2020-11-27 | The Peptide having Excellent Antioxidant Activity and Collagenase Inhibition Activity and Composition Containing the same |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| KR20220074281A true KR20220074281A (en) | 2022-06-03 |
| KR102506021B1 KR102506021B1 (en) | 2023-03-06 |
Family
ID=81982622
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| KR1020200162593A KR102506021B1 (en) | 2020-11-27 | 2020-11-27 | The Peptide having Excellent Antioxidant Activity and Collagenase Inhibition Activity and Composition Containing the same |
Country Status (1)
| Country | Link |
|---|---|
| KR (1) | KR102506021B1 (en) |
Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| KR20130113996A (en) * | 2012-04-06 | 2013-10-16 | 경남과학기술대학교 산학협력단 | Novel peptide with collagenase inhibitory activity and use thereof |
| KR20140026275A (en) * | 2012-08-24 | 2014-03-05 | 경상대학교산학협력단 | Novel peptide derived from oyster hydrolysate with collagenase inhibitory activity and their application |
| KR20160147293A (en) * | 2015-06-15 | 2016-12-23 | 이정복 | Skin external application composition and cosmetic composition comprising peptide derived silk cocoon or its derivatives with an anti-aging, antioxidant or anti-inflammatory effect |
-
2020
- 2020-11-27 KR KR1020200162593A patent/KR102506021B1/en active IP Right Grant
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| KR20130113996A (en) * | 2012-04-06 | 2013-10-16 | 경남과학기술대학교 산학협력단 | Novel peptide with collagenase inhibitory activity and use thereof |
| KR20140026275A (en) * | 2012-08-24 | 2014-03-05 | 경상대학교산학협력단 | Novel peptide derived from oyster hydrolysate with collagenase inhibitory activity and their application |
| KR20160147293A (en) * | 2015-06-15 | 2016-12-23 | 이정복 | Skin external application composition and cosmetic composition comprising peptide derived silk cocoon or its derivatives with an anti-aging, antioxidant or anti-inflammatory effect |
Also Published As
| Publication number | Publication date |
|---|---|
| KR102506021B1 (en) | 2023-03-06 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| KR20120105450A (en) | Peptides used in the treatment and/or care of the skin, mucous membranes and/or hair and its use in cosmetic or pharmaceutical compositions | |
| KR101763933B1 (en) | Phenolic acid derivative and cosmetic composition comprising the same | |
| CN110713517A (en) | Hyaluronan oligopeptide and preparation and application methods thereof | |
| KR101261065B1 (en) | Synthetic peptides reducing or removing bags formed under lower eye contour and their use in cosmetic or dermopharmaceutical compositions | |
| KR102506021B1 (en) | The Peptide having Excellent Antioxidant Activity and Collagenase Inhibition Activity and Composition Containing the same | |
| KR101667383B1 (en) | Biotin-conjugated hexapeptide-2 analogue and use thereof | |
| AU2020206415A1 (en) | Peptides and compositions for use in cosmetics | |
| KR102506020B1 (en) | The Peptide having Excellent Antioxidant Activity and Tyrosinase Inhibition Activity and Composition Containing the same | |
| KR102483429B1 (en) | Heptapeptide having Anti-inflammatory, Antioxidant or Collagen proliferation efficacy and Composition comprising the same | |
| KR102462608B1 (en) | Caffeoyl-tetrapeptide having anti-inflammatory and skin soothing effects and composition containing the same | |
| KR101343752B1 (en) | Novel Whitening compound and Whitening cosmetic composition comprising the same | |
| KR20240072327A (en) | A peptide having an excellent collagen proliferation effect and a composition for improving skin wrinkles comprising the same | |
| KR20230092139A (en) | The Octapeptide Having Melanin Production Inhibitory Activity and Composition Containing the same | |
| JP5123683B2 (en) | New peptide | |
| KR20240072326A (en) | A peptide having excellent skin cell regeneration effect and a composition for promoting skin cell regeneration comprising the same | |
| KR101224809B1 (en) | Retinoic acid derivative, process for preparing the same, and cosmetic composition comprising the same | |
| CN110520162B (en) | Combination of salicylic acid and peptide | |
| KR101942844B1 (en) | Gallic acid derivative, method for production thereof and external skin composition containing the same | |
| KR101945946B1 (en) | Gentisic acid derivative, method for production thereof and external skin composition containing the same | |
| KR101945947B1 (en) | Protocatechuic acid derivative, method for production thereof and external skin composition containing the same | |
| KR20240072372A (en) | Peptidomimetics having excellent itch-inducing cytokine control effect and cosmetic composition comprising the same | |
| KR20190128842A (en) | Manufacturing method and application of peptide derivatives with excellent effectiveness for collagen synthesis promotion and collagenase inhibition | |
| KR102120981B1 (en) | Gentisic acid derivative, method for production thereof and external skin composition containing the same | |
| KR20090132911A (en) | Peptides having activities of transforming growth factor-beta and their uses | |
| KR102660375B1 (en) | PGC-1α-derived Peptide Derivative and Cosmetic Composition Comprising the Same |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| E902 | Notification of reason for refusal | ||
| E701 | Decision to grant or registration of patent right | ||
| GRNT | Written decision to grant |