CN108714111A - A kind of cell-penetrating peptide-acetyl group Argireline nano-emulsion and preparation method thereof - Google Patents
A kind of cell-penetrating peptide-acetyl group Argireline nano-emulsion and preparation method thereof Download PDFInfo
- Publication number
- CN108714111A CN108714111A CN201810461431.6A CN201810461431A CN108714111A CN 108714111 A CN108714111 A CN 108714111A CN 201810461431 A CN201810461431 A CN 201810461431A CN 108714111 A CN108714111 A CN 108714111A
- Authority
- CN
- China
- Prior art keywords
- argireline
- cell
- peptide
- acetyl group
- penetrating peptide
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Granted
Links
- 108010006338 acetyl-glutamyl-glutamyl-methionyl-glutaminyl-arginyl-argininamide Proteins 0.000 title claims abstract description 131
- RJZNPROJTJSYLC-LLINQDLYSA-N (4s)-4-acetamido-5-[[(2s)-1-[[(2s)-1-[[(2s)-5-amino-1-[[(2s)-1-[[(2s)-1-amino-5-(diaminomethylideneamino)-1-oxopentan-2-yl]amino]-5-(diaminomethylideneamino)-1-oxopentan-2-yl]amino]-1,5-dioxopentan-2-yl]amino]-4-methylsulfanyl-1-oxobutan-2-yl]amino]-4-car Chemical compound OC(=O)CC[C@H](NC(C)=O)C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H](CCSC)C(=O)N[C@@H](CCC(N)=O)C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CCCNC(N)=N)C(N)=O RJZNPROJTJSYLC-LLINQDLYSA-N 0.000 title claims abstract description 122
- 239000007908 nanoemulsion Substances 0.000 title claims abstract description 91
- 238000002360 preparation method Methods 0.000 title claims abstract description 39
- 108090000765 processed proteins & peptides Proteins 0.000 claims abstract description 145
- 108010051109 Cell-Penetrating Peptides Proteins 0.000 claims abstract description 61
- 102000020313 Cell-Penetrating Peptides Human genes 0.000 claims abstract description 61
- 125000002777 acetyl group Chemical group [H]C([H])([H])C(*)=O 0.000 claims abstract description 42
- 238000000034 method Methods 0.000 claims abstract description 25
- 238000013461 design Methods 0.000 claims abstract description 14
- 230000004224 protection Effects 0.000 claims abstract description 7
- 238000010438 heat treatment Methods 0.000 claims abstract description 4
- ZMXDDKWLCZADIW-UHFFFAOYSA-N N,N-Dimethylformamide Chemical compound CN(C)C=O ZMXDDKWLCZADIW-UHFFFAOYSA-N 0.000 claims description 71
- 229920005989 resin Polymers 0.000 claims description 69
- 239000011347 resin Substances 0.000 claims description 69
- WEVYAHXRMPXWCK-UHFFFAOYSA-N Acetonitrile Chemical compound CC#N WEVYAHXRMPXWCK-UHFFFAOYSA-N 0.000 claims description 57
- YMWUJEATGCHHMB-UHFFFAOYSA-N Dichloromethane Chemical compound ClCCl YMWUJEATGCHHMB-UHFFFAOYSA-N 0.000 claims description 51
- 239000012071 phase Substances 0.000 claims description 44
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 42
- 150000001413 amino acids Chemical class 0.000 claims description 41
- 229940024606 amino acid Drugs 0.000 claims description 34
- 238000006243 chemical reaction Methods 0.000 claims description 29
- 239000007788 liquid Substances 0.000 claims description 24
- 239000000203 mixture Substances 0.000 claims description 23
- -1 Aliphatic ester Chemical class 0.000 claims description 22
- JUJWROOIHBZHMG-UHFFFAOYSA-N Pyridine Chemical compound C1=CC=NC=C1 JUJWROOIHBZHMG-UHFFFAOYSA-N 0.000 claims description 20
- 230000015572 biosynthetic process Effects 0.000 claims description 19
- 238000004128 high performance liquid chromatography Methods 0.000 claims description 19
- OKKJLVBELUTLKV-UHFFFAOYSA-N Methanol Chemical class OC OKKJLVBELUTLKV-UHFFFAOYSA-N 0.000 claims description 17
- 239000000047 product Substances 0.000 claims description 15
- RTZKZFJDLAIYFH-UHFFFAOYSA-N Diethyl ether Chemical compound CCOCC RTZKZFJDLAIYFH-UHFFFAOYSA-N 0.000 claims description 14
- 238000003786 synthesis reaction Methods 0.000 claims description 14
- 238000010511 deprotection reaction Methods 0.000 claims description 12
- JGFZNNIVVJXRND-UHFFFAOYSA-N N,N-Diisopropylethylamine (DIPEA) Chemical compound CCN(C(C)C)C(C)C JGFZNNIVVJXRND-UHFFFAOYSA-N 0.000 claims description 11
- 238000004458 analytical method Methods 0.000 claims description 11
- 125000003963 dichloro group Chemical group Cl* 0.000 claims description 11
- 239000012043 crude product Substances 0.000 claims description 10
- 238000005520 cutting process Methods 0.000 claims description 10
- 235000014113 dietary fatty acids Nutrition 0.000 claims description 10
- 229930195729 fatty acid Natural products 0.000 claims description 10
- 239000000194 fatty acid Substances 0.000 claims description 10
- UMJSCPRVCHMLSP-UHFFFAOYSA-N pyridine Natural products COC1=CC=CN=C1 UMJSCPRVCHMLSP-UHFFFAOYSA-N 0.000 claims description 10
- 238000005406 washing Methods 0.000 claims description 10
- WFDIJRYMOXRFFG-UHFFFAOYSA-N Acetic anhydride Chemical compound CC(=O)OC(C)=O WFDIJRYMOXRFFG-UHFFFAOYSA-N 0.000 claims description 9
- IJGRMHOSHXDMSA-UHFFFAOYSA-N Atomic nitrogen Chemical compound N#N IJGRMHOSHXDMSA-UHFFFAOYSA-N 0.000 claims description 9
- 239000002994 raw material Substances 0.000 claims description 9
- HIQIXEFWDLTDED-UHFFFAOYSA-N 4-hydroxy-1-piperidin-4-ylpyrrolidin-2-one Chemical compound O=C1CC(O)CN1C1CCNCC1 HIQIXEFWDLTDED-UHFFFAOYSA-N 0.000 claims description 8
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 claims description 8
- ISWSIDIOOBJBQZ-UHFFFAOYSA-N Phenol Chemical compound OC1=CC=CC=C1 ISWSIDIOOBJBQZ-UHFFFAOYSA-N 0.000 claims description 8
- 230000008602 contraction Effects 0.000 claims description 8
- 125000003088 (fluoren-9-ylmethoxy)carbonyl group Chemical group 0.000 claims description 7
- 230000008859 change Effects 0.000 claims description 7
- 125000006239 protecting group Chemical group 0.000 claims description 7
- 239000000126 substance Substances 0.000 claims description 7
- HNICLNKVURBTKV-NDEPHWFRSA-N (2s)-5-[[amino-[(2,2,4,6,7-pentamethyl-3h-1-benzofuran-5-yl)sulfonylamino]methylidene]amino]-2-(9h-fluoren-9-ylmethoxycarbonylamino)pentanoic acid Chemical compound C12=CC=CC=C2C2=CC=CC=C2C1COC(=O)N[C@H](C(O)=O)CCCN=C(N)NS(=O)(=O)C1=C(C)C(C)=C2OC(C)(C)CC2=C1C HNICLNKVURBTKV-NDEPHWFRSA-N 0.000 claims description 6
- NQRYJNQNLNOLGT-UHFFFAOYSA-N Piperidine Chemical compound C1CCNCC1 NQRYJNQNLNOLGT-UHFFFAOYSA-N 0.000 claims description 6
- 238000004140 cleaning Methods 0.000 claims description 6
- 239000012154 double-distilled water Substances 0.000 claims description 6
- 239000006166 lysate Substances 0.000 claims description 6
- 239000004475 Arginine Substances 0.000 claims description 5
- ODKSFYDXXFIFQN-UHFFFAOYSA-N arginine Natural products OC(=O)C(N)CCCNC(N)=N ODKSFYDXXFIFQN-UHFFFAOYSA-N 0.000 claims description 5
- 229960003121 arginine Drugs 0.000 claims description 5
- 235000013365 dairy product Nutrition 0.000 claims description 5
- 238000010828 elution Methods 0.000 claims description 5
- 235000019441 ethanol Nutrition 0.000 claims description 5
- 235000013336 milk Nutrition 0.000 claims description 5
- 239000008267 milk Substances 0.000 claims description 5
- 210000004080 milk Anatomy 0.000 claims description 5
- 229910052757 nitrogen Inorganic materials 0.000 claims description 5
- 150000003053 piperidines Chemical class 0.000 claims description 5
- 239000007858 starting material Substances 0.000 claims description 5
- 230000008961 swelling Effects 0.000 claims description 5
- 239000012153 distilled water Substances 0.000 claims description 4
- 238000003760 magnetic stirring Methods 0.000 claims description 4
- FEMOMIGRRWSMCU-UHFFFAOYSA-N ninhydrin Chemical compound C1=CC=C2C(=O)C(O)(O)C(=O)C2=C1 FEMOMIGRRWSMCU-UHFFFAOYSA-N 0.000 claims description 4
- 230000010355 oscillation Effects 0.000 claims description 4
- 239000007787 solid Substances 0.000 claims description 4
- 230000004913 activation Effects 0.000 claims description 3
- 238000005374 membrane filtration Methods 0.000 claims description 3
- 239000002904 solvent Substances 0.000 claims description 3
- 238000003756 stirring Methods 0.000 claims description 3
- 238000006467 substitution reaction Methods 0.000 claims description 3
- 239000006228 supernatant Substances 0.000 claims description 3
- 238000002604 ultrasonography Methods 0.000 claims description 3
- 238000013019 agitation Methods 0.000 claims description 2
- 238000001914 filtration Methods 0.000 claims description 2
- 239000007790 solid phase Substances 0.000 claims description 2
- 238000010189 synthetic method Methods 0.000 claims description 2
- 230000002378 acidificating effect Effects 0.000 claims 1
- 230000000694 effects Effects 0.000 abstract description 64
- 239000003814 drug Substances 0.000 abstract description 45
- 238000010521 absorption reaction Methods 0.000 abstract description 44
- 229940079593 drug Drugs 0.000 abstract description 40
- 239000002537 cosmetic Substances 0.000 abstract description 29
- 230000003796 beauty Effects 0.000 abstract description 27
- 230000003712 anti-aging effect Effects 0.000 abstract description 5
- 230000015556 catabolic process Effects 0.000 abstract description 5
- 238000006731 degradation reaction Methods 0.000 abstract description 4
- 230000004048 modification Effects 0.000 abstract description 4
- 238000012986 modification Methods 0.000 abstract description 4
- 206010020751 Hypersensitivity Diseases 0.000 abstract description 2
- 230000036541 health Effects 0.000 abstract description 2
- 230000002195 synergetic effect Effects 0.000 abstract description 2
- 208000026935 allergic disease Diseases 0.000 abstract 1
- 230000007815 allergy Effects 0.000 abstract 1
- 230000007613 environmental effect Effects 0.000 abstract 1
- 238000009434 installation Methods 0.000 abstract 1
- 238000004519 manufacturing process Methods 0.000 abstract 1
- 230000007903 penetration ability Effects 0.000 abstract 1
- 210000003491 skin Anatomy 0.000 description 100
- 102000004196 processed proteins & peptides Human genes 0.000 description 51
- 230000035515 penetration Effects 0.000 description 40
- 229920001184 polypeptide Polymers 0.000 description 38
- 235000001014 amino acid Nutrition 0.000 description 30
- 230000006870 function Effects 0.000 description 25
- 239000004530 micro-emulsion Substances 0.000 description 21
- DTQVDTLACAAQTR-UHFFFAOYSA-N Trifluoroacetic acid Chemical compound OC(=O)C(F)(F)F DTQVDTLACAAQTR-UHFFFAOYSA-N 0.000 description 20
- 235000018102 proteins Nutrition 0.000 description 20
- 102000004169 proteins and genes Human genes 0.000 description 20
- 108090000623 proteins and genes Proteins 0.000 description 20
- 238000005516 engineering process Methods 0.000 description 17
- 239000003995 emulsifying agent Substances 0.000 description 14
- 210000004027 cell Anatomy 0.000 description 13
- 230000037303 wrinkles Effects 0.000 description 13
- 230000021736 acetylation Effects 0.000 description 12
- 238000006640 acetylation reaction Methods 0.000 description 12
- 210000002615 epidermis Anatomy 0.000 description 12
- 230000007246 mechanism Effects 0.000 description 12
- 230000031018 biological processes and functions Effects 0.000 description 11
- 239000003921 oil Substances 0.000 description 11
- 235000019198 oils Nutrition 0.000 description 11
- 239000000243 solution Substances 0.000 description 11
- 210000000170 cell membrane Anatomy 0.000 description 10
- 230000008569 process Effects 0.000 description 10
- 238000011160 research Methods 0.000 description 10
- 239000002253 acid Substances 0.000 description 9
- 239000004064 cosurfactant Substances 0.000 description 9
- 239000004615 ingredient Substances 0.000 description 9
- 239000004094 surface-active agent Substances 0.000 description 9
- 238000013459 approach Methods 0.000 description 8
- 150000002632 lipids Chemical class 0.000 description 8
- 210000003205 muscle Anatomy 0.000 description 8
- 210000005036 nerve Anatomy 0.000 description 8
- 210000001519 tissue Anatomy 0.000 description 8
- 230000008901 benefit Effects 0.000 description 7
- 238000009833 condensation Methods 0.000 description 7
- 230000005494 condensation Effects 0.000 description 7
- 230000007547 defect Effects 0.000 description 7
- 239000002585 base Substances 0.000 description 6
- 230000004071 biological effect Effects 0.000 description 6
- 235000013339 cereals Nutrition 0.000 description 6
- 229910052801 chlorine Inorganic materials 0.000 description 6
- 239000000460 chlorine Substances 0.000 description 6
- 239000003349 gelling agent Substances 0.000 description 6
- 238000009740 moulding (composite fabrication) Methods 0.000 description 6
- 238000012360 testing method Methods 0.000 description 6
- 108030001720 Bontoxilysin Proteins 0.000 description 5
- ZAMOUSCENKQFHK-UHFFFAOYSA-N Chlorine atom Chemical compound [Cl] ZAMOUSCENKQFHK-UHFFFAOYSA-N 0.000 description 5
- 230000000975 bioactive effect Effects 0.000 description 5
- 229940053031 botulinum toxin Drugs 0.000 description 5
- 239000003795 chemical substances by application Substances 0.000 description 5
- 210000000736 corneocyte Anatomy 0.000 description 5
- 239000000839 emulsion Substances 0.000 description 5
- 210000001097 facial muscle Anatomy 0.000 description 5
- 238000005259 measurement Methods 0.000 description 5
- 125000000913 palmityl group Chemical group [H]C([*])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])[H] 0.000 description 5
- 239000002245 particle Substances 0.000 description 5
- QGZKDVFQNNGYKY-UHFFFAOYSA-N Ammonia Chemical compound N QGZKDVFQNNGYKY-UHFFFAOYSA-N 0.000 description 4
- IAZDPXIOMUYVGZ-UHFFFAOYSA-N Dimethylsulphoxide Chemical compound CS(C)=O IAZDPXIOMUYVGZ-UHFFFAOYSA-N 0.000 description 4
- 239000000232 Lipid Bilayer Substances 0.000 description 4
- GLUUGHFHXGJENI-UHFFFAOYSA-N Piperazine Chemical compound C1CNCCN1 GLUUGHFHXGJENI-UHFFFAOYSA-N 0.000 description 4
- 230000009471 action Effects 0.000 description 4
- 125000002252 acyl group Chemical group 0.000 description 4
- 230000001153 anti-wrinkle effect Effects 0.000 description 4
- 125000003178 carboxy group Chemical group [H]OC(*)=O 0.000 description 4
- 230000007423 decrease Effects 0.000 description 4
- 238000001514 detection method Methods 0.000 description 4
- 239000006185 dispersion Substances 0.000 description 4
- 238000009826 distribution Methods 0.000 description 4
- 210000003780 hair follicle Anatomy 0.000 description 4
- 230000004118 muscle contraction Effects 0.000 description 4
- 206010033675 panniculitis Diseases 0.000 description 4
- 230000035790 physiological processes and functions Effects 0.000 description 4
- 125000002924 primary amino group Chemical group [H]N([H])* 0.000 description 4
- 230000003252 repetitive effect Effects 0.000 description 4
- 210000001732 sebaceous gland Anatomy 0.000 description 4
- 230000003068 static effect Effects 0.000 description 4
- 210000004304 subcutaneous tissue Anatomy 0.000 description 4
- 210000000106 sweat gland Anatomy 0.000 description 4
- 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 4
- 238000005303 weighing Methods 0.000 description 4
- AJLNZWYOJAWBCR-OOPVGHQCSA-N (4s)-4-acetamido-5-[[(2s)-1-[[(2s)-1-[[(2s)-5-amino-1-[[(2s)-1-[[(2s)-1-amino-5-(diaminomethylideneamino)-1-oxopentan-2-yl]amino]-5-(diaminomethylideneamino)-1-oxopentan-2-yl]amino]-1,5-dioxopentan-2-yl]amino]-4-methylsulfanyl-1-oxobutan-2-yl]amino]-4-car Chemical compound OC(=O)CC[C@H](NC(C)=O)C(=C)N[C@@H](CCC(O)=O)C(=O)N[C@@H](CCSC)C(=O)N[C@@H](CCC(N)=O)C(=O)N[C@@H](CCCN=C(N)N)C(=O)N[C@@H](CCCN=C(N)N)C(N)=O AJLNZWYOJAWBCR-OOPVGHQCSA-N 0.000 description 3
- UHOVQNZJYSORNB-UHFFFAOYSA-N Benzene Chemical compound C1=CC=CC=C1 UHOVQNZJYSORNB-UHFFFAOYSA-N 0.000 description 3
- 102000005853 Clathrin Human genes 0.000 description 3
- 108010019874 Clathrin Proteins 0.000 description 3
- PEDCQBHIVMGVHV-UHFFFAOYSA-N Glycerine Chemical compound OCC(O)CO PEDCQBHIVMGVHV-UHFFFAOYSA-N 0.000 description 3
- 206010020649 Hyperkeratosis Diseases 0.000 description 3
- 208000001126 Keratosis Diseases 0.000 description 3
- KDXKERNSBIXSRK-UHFFFAOYSA-N Lysine Natural products NCCCCC(N)C(O)=O KDXKERNSBIXSRK-UHFFFAOYSA-N 0.000 description 3
- 239000004472 Lysine Substances 0.000 description 3
- 108010038807 Oligopeptides Proteins 0.000 description 3
- 102000015636 Oligopeptides Human genes 0.000 description 3
- 238000007792 addition Methods 0.000 description 3
- 239000003153 chemical reaction reagent Substances 0.000 description 3
- 229930193282 clathrin Natural products 0.000 description 3
- 229910052802 copper Inorganic materials 0.000 description 3
- 239000010949 copper Substances 0.000 description 3
- 210000000805 cytoplasm Anatomy 0.000 description 3
- 230000007812 deficiency Effects 0.000 description 3
- 230000012202 endocytosis Effects 0.000 description 3
- 239000003623 enhancer Substances 0.000 description 3
- 230000001815 facial effect Effects 0.000 description 3
- 239000000835 fiber Substances 0.000 description 3
- 210000004209 hair Anatomy 0.000 description 3
- 238000002347 injection Methods 0.000 description 3
- 239000007924 injection Substances 0.000 description 3
- 230000003834 intracellular effect Effects 0.000 description 3
- 239000010985 leather Substances 0.000 description 3
- 239000007791 liquid phase Substances 0.000 description 3
- 235000013372 meat Nutrition 0.000 description 3
- 230000010534 mechanism of action Effects 0.000 description 3
- 230000007830 nerve conduction Effects 0.000 description 3
- 108020004707 nucleic acids Proteins 0.000 description 3
- 102000039446 nucleic acids Human genes 0.000 description 3
- 150000007523 nucleic acids Chemical class 0.000 description 3
- 210000000056 organ Anatomy 0.000 description 3
- 230000035699 permeability Effects 0.000 description 3
- 239000012466 permeate Substances 0.000 description 3
- 229920000724 poly(L-arginine) polymer Polymers 0.000 description 3
- 108010011110 polyarginine Proteins 0.000 description 3
- 230000002829 reductive effect Effects 0.000 description 3
- 230000008591 skin barrier function Effects 0.000 description 3
- 238000010532 solid phase synthesis reaction Methods 0.000 description 3
- 230000002269 spontaneous effect Effects 0.000 description 3
- 238000010361 transduction Methods 0.000 description 3
- 230000026683 transduction Effects 0.000 description 3
- 230000032258 transport Effects 0.000 description 3
- WSGCRSMLXFHGRM-DEVHWETNSA-N (2s)-2-[[(2s)-6-amino-2-[[(2s,3r)-2-[[(2s,3r)-2-[[(2s)-6-amino-2-(hexadecanoylamino)hexanoyl]amino]-3-hydroxybutanoyl]amino]-3-hydroxybutanoyl]amino]hexanoyl]amino]-3-hydroxypropanoic acid Chemical group CCCCCCCCCCCCCCCC(=O)N[C@@H](CCCCN)C(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CCCCN)C(=O)N[C@@H](CO)C(O)=O WSGCRSMLXFHGRM-DEVHWETNSA-N 0.000 description 2
- 108010088751 Albumins Proteins 0.000 description 2
- 102000009027 Albumins Human genes 0.000 description 2
- RYGMFSIKBFXOCR-UHFFFAOYSA-N Copper Chemical compound [Cu] RYGMFSIKBFXOCR-UHFFFAOYSA-N 0.000 description 2
- PMATZTZNYRCHOR-CGLBZJNRSA-N Cyclosporin A Chemical compound CC[C@@H]1NC(=O)[C@H]([C@H](O)[C@H](C)C\C=C\C)N(C)C(=O)[C@H](C(C)C)N(C)C(=O)[C@H](CC(C)C)N(C)C(=O)[C@H](CC(C)C)N(C)C(=O)[C@@H](C)NC(=O)[C@H](C)NC(=O)[C@H](CC(C)C)N(C)C(=O)[C@H](C(C)C)NC(=O)[C@H](CC(C)C)N(C)C(=O)CN(C)C1=O PMATZTZNYRCHOR-CGLBZJNRSA-N 0.000 description 2
- 229930105110 Cyclosporin A Natural products 0.000 description 2
- 108010036949 Cyclosporine Proteins 0.000 description 2
- 108010016626 Dipeptides Proteins 0.000 description 2
- MVORZMQFXBLMHM-QWRGUYRKSA-N Gly-His-Lys Chemical compound NCCCC[C@@H](C(O)=O)NC(=O)[C@@H](NC(=O)CN)CC1=CN=CN1 MVORZMQFXBLMHM-QWRGUYRKSA-N 0.000 description 2
- DHMQDGOQFOQNFH-UHFFFAOYSA-N Glycine Chemical compound NCC(O)=O DHMQDGOQFOQNFH-UHFFFAOYSA-N 0.000 description 2
- 206010061218 Inflammation Diseases 0.000 description 2
- 102000011782 Keratins Human genes 0.000 description 2
- 108010076876 Keratins Proteins 0.000 description 2
- 208000005374 Poisoning Diseases 0.000 description 2
- 239000002202 Polyethylene glycol Substances 0.000 description 2
- 206010040880 Skin irritation Diseases 0.000 description 2
- 108010057266 Type A Botulinum Toxins Proteins 0.000 description 2
- OIPILFWXSMYKGL-UHFFFAOYSA-N acetylcholine Chemical compound CC(=O)OCC[N+](C)(C)C OIPILFWXSMYKGL-UHFFFAOYSA-N 0.000 description 2
- 229960004373 acetylcholine Drugs 0.000 description 2
- 230000001154 acute effect Effects 0.000 description 2
- 239000000654 additive Substances 0.000 description 2
- 230000000996 additive effect Effects 0.000 description 2
- 230000002776 aggregation Effects 0.000 description 2
- 238000004220 aggregation Methods 0.000 description 2
- 239000003513 alkali Substances 0.000 description 2
- 150000001408 amides Chemical group 0.000 description 2
- 125000000539 amino acid group Chemical group 0.000 description 2
- 229910021529 ammonia Inorganic materials 0.000 description 2
- 239000007864 aqueous solution Substances 0.000 description 2
- 230000004888 barrier function Effects 0.000 description 2
- 230000009286 beneficial effect Effects 0.000 description 2
- 230000005540 biological transmission Effects 0.000 description 2
- 210000004204 blood vessel Anatomy 0.000 description 2
- 229940094657 botulinum toxin type a Drugs 0.000 description 2
- 210000004899 c-terminal region Anatomy 0.000 description 2
- 150000003943 catecholamines Chemical class 0.000 description 2
- HVYWMOMLDIMFJA-DPAQBDIFSA-N cholesterol Chemical compound C1C=C2C[C@@H](O)CC[C@]2(C)[C@@H]2[C@@H]1[C@@H]1CC[C@H]([C@H](C)CCCC(C)C)[C@@]1(C)CC2 HVYWMOMLDIMFJA-DPAQBDIFSA-N 0.000 description 2
- 229960001265 ciclosporin Drugs 0.000 description 2
- 150000001875 compounds Chemical class 0.000 description 2
- 239000000470 constituent Substances 0.000 description 2
- 229930182912 cyclosporin Natural products 0.000 description 2
- 230000006378 damage Effects 0.000 description 2
- 238000005202 decontamination Methods 0.000 description 2
- 230000003588 decontaminative effect Effects 0.000 description 2
- 230000007123 defense Effects 0.000 description 2
- 210000004177 elastic tissue Anatomy 0.000 description 2
- 238000002330 electrospray ionisation mass spectrometry Methods 0.000 description 2
- 230000009881 electrostatic interaction Effects 0.000 description 2
- 230000002708 enhancing effect Effects 0.000 description 2
- 210000002919 epithelial cell Anatomy 0.000 description 2
- 150000002148 esters Chemical class 0.000 description 2
- 238000002474 experimental method Methods 0.000 description 2
- 230000008921 facial expression Effects 0.000 description 2
- 238000000855 fermentation Methods 0.000 description 2
- 230000004151 fermentation Effects 0.000 description 2
- 210000001061 forehead Anatomy 0.000 description 2
- 238000009472 formulation Methods 0.000 description 2
- 239000012634 fragment Substances 0.000 description 2
- 125000000524 functional group Chemical group 0.000 description 2
- 229910052736 halogen Inorganic materials 0.000 description 2
- 150000002367 halogens Chemical class 0.000 description 2
- 210000004276 hyalin Anatomy 0.000 description 2
- 230000007062 hydrolysis Effects 0.000 description 2
- 238000006460 hydrolysis reaction Methods 0.000 description 2
- 230000002209 hydrophobic effect Effects 0.000 description 2
- 238000001727 in vivo Methods 0.000 description 2
- 230000008595 infiltration Effects 0.000 description 2
- 238000001764 infiltration Methods 0.000 description 2
- 230000004054 inflammatory process Effects 0.000 description 2
- 230000002401 inhibitory effect Effects 0.000 description 2
- NOESYZHRGYRDHS-UHFFFAOYSA-N insulin Chemical compound N1C(=O)C(NC(=O)C(CCC(N)=O)NC(=O)C(CCC(O)=O)NC(=O)C(C(C)C)NC(=O)C(NC(=O)CN)C(C)CC)CSSCC(C(NC(CO)C(=O)NC(CC(C)C)C(=O)NC(CC=2C=CC(O)=CC=2)C(=O)NC(CCC(N)=O)C(=O)NC(CC(C)C)C(=O)NC(CCC(O)=O)C(=O)NC(CC(N)=O)C(=O)NC(CC=2C=CC(O)=CC=2)C(=O)NC(CSSCC(NC(=O)C(C(C)C)NC(=O)C(CC(C)C)NC(=O)C(CC=2C=CC(O)=CC=2)NC(=O)C(CC(C)C)NC(=O)C(C)NC(=O)C(CCC(O)=O)NC(=O)C(C(C)C)NC(=O)C(CC(C)C)NC(=O)C(CC=2NC=NC=2)NC(=O)C(CO)NC(=O)CNC2=O)C(=O)NCC(=O)NC(CCC(O)=O)C(=O)NC(CCCNC(N)=N)C(=O)NCC(=O)NC(CC=3C=CC=CC=3)C(=O)NC(CC=3C=CC=CC=3)C(=O)NC(CC=3C=CC(O)=CC=3)C(=O)NC(C(C)O)C(=O)N3C(CCC3)C(=O)NC(CCCCN)C(=O)NC(C)C(O)=O)C(=O)NC(CC(N)=O)C(O)=O)=O)NC(=O)C(C(C)CC)NC(=O)C(CO)NC(=O)C(C(C)O)NC(=O)C1CSSCC2NC(=O)C(CC(C)C)NC(=O)C(NC(=O)C(CCC(N)=O)NC(=O)C(CC(N)=O)NC(=O)C(NC(=O)C(N)CC=1C=CC=CC=1)C(C)C)CC1=CN=CN1 NOESYZHRGYRDHS-UHFFFAOYSA-N 0.000 description 2
- 239000013067 intermediate product Substances 0.000 description 2
- 239000002502 liposome Substances 0.000 description 2
- 210000004962 mammalian cell Anatomy 0.000 description 2
- 239000000463 material Substances 0.000 description 2
- 239000012528 membrane Substances 0.000 description 2
- 230000004060 metabolic process Effects 0.000 description 2
- 150000002825 nitriles Chemical class 0.000 description 2
- 230000000474 nursing effect Effects 0.000 description 2
- 235000016709 nutrition Nutrition 0.000 description 2
- 230000010494 opalescence Effects 0.000 description 2
- 230000003204 osmotic effect Effects 0.000 description 2
- 230000036407 pain Effects 0.000 description 2
- 125000001312 palmitoyl group Chemical group O=C([*])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])[H] 0.000 description 2
- 208000035824 paresthesia Diseases 0.000 description 2
- 238000010647 peptide synthesis reaction Methods 0.000 description 2
- 230000000144 pharmacologic effect Effects 0.000 description 2
- 230000008884 pinocytosis Effects 0.000 description 2
- 231100000572 poisoning Toxicity 0.000 description 2
- 230000000607 poisoning effect Effects 0.000 description 2
- 229920001223 polyethylene glycol Polymers 0.000 description 2
- 229920000642 polymer Polymers 0.000 description 2
- 238000003825 pressing Methods 0.000 description 2
- 238000012827 research and development Methods 0.000 description 2
- 230000002441 reversible effect Effects 0.000 description 2
- 238000009738 saturating Methods 0.000 description 2
- 230000009759 skin aging Effects 0.000 description 2
- 230000036556 skin irritation Effects 0.000 description 2
- 231100000475 skin irritation Toxicity 0.000 description 2
- 230000001954 sterilising effect Effects 0.000 description 2
- 230000000638 stimulation Effects 0.000 description 2
- 210000000498 stratum granulosum Anatomy 0.000 description 2
- 210000000437 stratum spinosum Anatomy 0.000 description 2
- 239000003053 toxin Substances 0.000 description 2
- 231100000765 toxin Toxicity 0.000 description 2
- 238000000825 ultraviolet detection Methods 0.000 description 2
- OGNHOGPWQTWKGQ-VGWMRTNUSA-N (2S)-5-amino-2-[[(2S)-1-[2-[[(2S)-5-amino-2-[[(2S)-1-(2-aminoacetyl)pyrrolidine-2-carbonyl]amino]-5-oxopentanoyl]amino]acetyl]pyrrolidine-2-carbonyl]amino]-5-oxopentanoic acid Chemical compound NCC(=O)N1CCC[C@H]1C(=O)N[C@@H](CCC(N)=O)C(=O)NCC(=O)N1[C@H](C(=O)N[C@@H](CCC(N)=O)C(O)=O)CCC1 OGNHOGPWQTWKGQ-VGWMRTNUSA-N 0.000 description 1
- JAUKCFULLJFBFN-VWLOTQADSA-N (2s)-2-(9h-fluoren-9-ylmethoxycarbonylamino)-3-[4-[(2-methylpropan-2-yl)oxy]phenyl]propanoic acid Chemical compound C1=CC(OC(C)(C)C)=CC=C1C[C@@H](C(O)=O)NC(=O)OCC1C2=CC=CC=C2C2=CC=CC=C21 JAUKCFULLJFBFN-VWLOTQADSA-N 0.000 description 1
- BUBGAUHBELNDEW-SFHVURJKSA-N (2s)-2-(9h-fluoren-9-ylmethoxycarbonylamino)-4-methylsulfanylbutanoic acid Chemical compound C1=CC=C2C(COC(=O)N[C@@H](CCSC)C(O)=O)C3=CC=CC=C3C2=C1 BUBGAUHBELNDEW-SFHVURJKSA-N 0.000 description 1
- WDGICUODAOGOMO-DHUJRADRSA-N (2s)-2-(9h-fluoren-9-ylmethoxycarbonylamino)-5-oxo-5-(tritylamino)pentanoic acid Chemical compound C([C@@H](C(=O)O)NC(=O)OCC1C2=CC=CC=C2C2=CC=CC=C21)CC(=O)NC(C=1C=CC=CC=1)(C=1C=CC=CC=1)C1=CC=CC=C1 WDGICUODAOGOMO-DHUJRADRSA-N 0.000 description 1
- UMRUUWFGLGNQLI-QFIPXVFZSA-N (2s)-2-(9h-fluoren-9-ylmethoxycarbonylamino)-6-[(2-methylpropan-2-yl)oxycarbonylamino]hexanoic acid Chemical compound C1=CC=C2C(COC(=O)N[C@@H](CCCCNC(=O)OC(C)(C)C)C(O)=O)C3=CC=CC=C3C2=C1 UMRUUWFGLGNQLI-QFIPXVFZSA-N 0.000 description 1
- QTEWJFPLRGWOOU-IHRRRGAJSA-N (2s)-2-[[(2s)-2-[[(2s)-2-(3-aminopropanoylamino)-3-(1h-imidazol-5-yl)propanoyl]amino]-3-hydroxypropanoyl]amino]-3-(1h-imidazol-5-yl)propanoic acid Chemical compound C([C@H](NC(=O)CCN)C(=O)N[C@@H](CO)C(=O)N[C@@H](CC=1NC=NC=1)C(O)=O)C1=CN=CN1 QTEWJFPLRGWOOU-IHRRRGAJSA-N 0.000 description 1
- OFGVZFQUFJYSGS-CPDXTSBQSA-N (2s)-2-[[(2s)-2-[[(2s)-2-[[(2s)-6-amino-2-[[(2s,3s)-2-[[(2s)-2-amino-3-hydroxypropanoyl]amino]-3-methylpentanoyl]amino]hexanoyl]amino]-3-methylbutanoyl]amino]propanoyl]amino]-3-methylbutanoic acid Chemical compound OC[C@H](N)C(=O)N[C@@H]([C@@H](C)CC)C(=O)N[C@@H](CCCCN)C(=O)N[C@@H](C(C)C)C(=O)N[C@@H](C)C(=O)N[C@@H](C(C)C)C(O)=O OFGVZFQUFJYSGS-CPDXTSBQSA-N 0.000 description 1
- DVBUCBXGDWWXNY-SFHVURJKSA-N (2s)-5-(diaminomethylideneamino)-2-(9h-fluoren-9-ylmethoxycarbonylamino)pentanoic acid Chemical compound C1=CC=C2C(COC(=O)N[C@@H](CCCN=C(N)N)C(O)=O)C3=CC=CC=C3C2=C1 DVBUCBXGDWWXNY-SFHVURJKSA-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
- NDKDFTQNXLHCGO-UHFFFAOYSA-N 2-(9h-fluoren-9-ylmethoxycarbonylamino)acetic acid Chemical compound C1=CC=C2C(COC(=O)NCC(=O)O)C3=CC=CC=C3C2=C1 NDKDFTQNXLHCGO-UHFFFAOYSA-N 0.000 description 1
- QCVGEOXPDFCNHA-UHFFFAOYSA-N 5,5-dimethyl-2,4-dioxo-1,3-oxazolidine-3-carboxamide Chemical compound CC1(C)OC(=O)N(C(N)=O)C1=O QCVGEOXPDFCNHA-UHFFFAOYSA-N 0.000 description 1
- 241000251468 Actinopterygii Species 0.000 description 1
- 206010002091 Anaesthesia Diseases 0.000 description 1
- 241000193830 Bacillus <bacterium> Species 0.000 description 1
- QRYRORQUOLYVBU-VBKZILBWSA-N Carnosic acid Natural products CC([C@@H]1CC2)(C)CCC[C@]1(C(O)=O)C1=C2C=C(C(C)C)C(O)=C1O QRYRORQUOLYVBU-VBKZILBWSA-N 0.000 description 1
- 108010087806 Carnosine Proteins 0.000 description 1
- 102000001187 Collagen Type III Human genes 0.000 description 1
- 108010069502 Collagen Type III Proteins 0.000 description 1
- 206010010904 Convulsion Diseases 0.000 description 1
- 102000002322 Egg Proteins Human genes 0.000 description 1
- 108010000912 Egg Proteins Proteins 0.000 description 1
- 241000196324 Embryophyta Species 0.000 description 1
- 102000004190 Enzymes Human genes 0.000 description 1
- 108090000790 Enzymes Proteins 0.000 description 1
- 239000001293 FEMA 3089 Substances 0.000 description 1
- 102000009123 Fibrin Human genes 0.000 description 1
- 108010073385 Fibrin Proteins 0.000 description 1
- BWGVNKXGVNDBDI-UHFFFAOYSA-N Fibrin monomer Chemical compound CNC(=O)CNC(=O)CN BWGVNKXGVNDBDI-UHFFFAOYSA-N 0.000 description 1
- WHUUTDBJXJRKMK-UHFFFAOYSA-N Glutamic acid Natural products OC(=O)C(N)CCC(O)=O WHUUTDBJXJRKMK-UHFFFAOYSA-N 0.000 description 1
- 239000004471 Glycine Substances 0.000 description 1
- 229920002683 Glycosaminoglycan Polymers 0.000 description 1
- 229920002971 Heparan sulfate Polymers 0.000 description 1
- HTTJABKRGRZYRN-UHFFFAOYSA-N Heparin Chemical compound OC1C(NC(=O)C)C(O)OC(COS(O)(=O)=O)C1OC1C(OS(O)(=O)=O)C(O)C(OC2C(C(OS(O)(=O)=O)C(OC3C(C(O)C(O)C(O3)C(O)=O)OS(O)(=O)=O)C(CO)O2)NS(O)(=O)=O)C(C(O)=O)O1 HTTJABKRGRZYRN-UHFFFAOYSA-N 0.000 description 1
- 102000004877 Insulin Human genes 0.000 description 1
- 108090001061 Insulin Proteins 0.000 description 1
- FFEARJCKVFRZRR-BYPYZUCNSA-N L-methionine Chemical compound CSCC[C@H](N)C(O)=O FFEARJCKVFRZRR-BYPYZUCNSA-N 0.000 description 1
- OUYCCCASQSFEME-QMMMGPOBSA-N L-tyrosine Chemical compound OC(=O)[C@@H](N)CC1=CC=C(O)C=C1 OUYCCCASQSFEME-QMMMGPOBSA-N 0.000 description 1
- 208000003351 Melanosis Diseases 0.000 description 1
- 241001465754 Metazoa Species 0.000 description 1
- 239000004909 Moisturizer Substances 0.000 description 1
- CQOVPNPJLQNMDC-UHFFFAOYSA-N N-beta-alanyl-L-histidine Natural products NCCC(=O)NC(C(O)=O)CC1=CN=CN1 CQOVPNPJLQNMDC-UHFFFAOYSA-N 0.000 description 1
- 101800001147 Octapeptide 1 Proteins 0.000 description 1
- 108091034117 Oligonucleotide Proteins 0.000 description 1
- 108091005804 Peptidases Proteins 0.000 description 1
- 235000008331 Pinus X rigitaeda Nutrition 0.000 description 1
- 235000011613 Pinus brutia Nutrition 0.000 description 1
- 241000018646 Pinus brutia Species 0.000 description 1
- 101710093543 Probable non-specific lipid-transfer protein Proteins 0.000 description 1
- 239000004365 Protease Substances 0.000 description 1
- 102000016611 Proteoglycans Human genes 0.000 description 1
- 108010067787 Proteoglycans Proteins 0.000 description 1
- 241000700159 Rattus Species 0.000 description 1
- 102100037486 Reverse transcriptase/ribonuclease H Human genes 0.000 description 1
- 206010053615 Thermal burn Diseases 0.000 description 1
- NPTLAYTZMHJJDP-KTKRTIGZSA-N [3-[3-[3-[3-[3-[3-[3-[3-[3-(2,3-dihydroxypropoxy)-2-hydroxypropoxy]-2-hydroxypropoxy]-2-hydroxypropoxy]-2-hydroxypropoxy]-2-hydroxypropoxy]-2-hydroxypropoxy]-2-hydroxypropoxy]-2-hydroxypropoxy]-2-hydroxypropyl] (z)-octadec-9-enoate Chemical compound CCCCCCCC\C=C/CCCCCCCC(=O)OCC(O)COCC(O)COCC(O)COCC(O)COCC(O)COCC(O)COCC(O)COCC(O)COCC(O)COCC(O)CO NPTLAYTZMHJJDP-KTKRTIGZSA-N 0.000 description 1
- 239000000370 acceptor Substances 0.000 description 1
- 150000004075 acetic anhydrides Chemical class 0.000 description 1
- 230000003213 activating effect Effects 0.000 description 1
- 210000001789 adipocyte Anatomy 0.000 description 1
- 230000032683 aging Effects 0.000 description 1
- 150000001298 alcohols Chemical class 0.000 description 1
- 125000001931 aliphatic group Chemical group 0.000 description 1
- 229930013930 alkaloid Natural products 0.000 description 1
- 208000030961 allergic reaction Diseases 0.000 description 1
- 230000004075 alteration Effects 0.000 description 1
- 229920003180 amino resin Polymers 0.000 description 1
- 239000003708 ampul Substances 0.000 description 1
- 230000037005 anaesthesia Effects 0.000 description 1
- 230000036592 analgesia Effects 0.000 description 1
- 230000003266 anti-allergic effect Effects 0.000 description 1
- 239000003963 antioxidant agent Substances 0.000 description 1
- 230000003078 antioxidant effect Effects 0.000 description 1
- 235000006708 antioxidants Nutrition 0.000 description 1
- 239000002249 anxiolytic agent Substances 0.000 description 1
- 125000000637 arginyl group Chemical group N[C@@H](CCCNC(N)=N)C(=O)* 0.000 description 1
- 239000000823 artificial membrane Substances 0.000 description 1
- 210000000270 basal cell Anatomy 0.000 description 1
- 230000008827 biological function Effects 0.000 description 1
- 230000033228 biological regulation Effects 0.000 description 1
- 230000008499 blood brain barrier function Effects 0.000 description 1
- 230000017531 blood circulation Effects 0.000 description 1
- 210000001218 blood-brain barrier Anatomy 0.000 description 1
- 229940044199 carnosine Drugs 0.000 description 1
- CQOVPNPJLQNMDC-ZETCQYMHSA-N carnosine Chemical compound [NH3+]CCC(=O)N[C@H](C([O-])=O)CC1=CNC=N1 CQOVPNPJLQNMDC-ZETCQYMHSA-N 0.000 description 1
- 150000001768 cations Chemical class 0.000 description 1
- 230000032823 cell division Effects 0.000 description 1
- 230000001413 cellular effect Effects 0.000 description 1
- 230000004700 cellular uptake Effects 0.000 description 1
- 230000003399 chemotactic effect Effects 0.000 description 1
- 125000001309 chloro group Chemical group Cl* 0.000 description 1
- 235000012000 cholesterol Nutrition 0.000 description 1
- 230000001713 cholinergic effect Effects 0.000 description 1
- 238000004587 chromatography analysis Methods 0.000 description 1
- 230000004087 circulation Effects 0.000 description 1
- 238000004581 coalescence Methods 0.000 description 1
- 239000011248 coating agent Substances 0.000 description 1
- 238000000576 coating method Methods 0.000 description 1
- 238000001246 colloidal dispersion Methods 0.000 description 1
- 230000000295 complement effect Effects 0.000 description 1
- 238000006482 condensation reaction Methods 0.000 description 1
- 235000009508 confectionery Nutrition 0.000 description 1
- 238000012790 confirmation Methods 0.000 description 1
- 210000002808 connective tissue Anatomy 0.000 description 1
- 230000001276 controlling effect Effects 0.000 description 1
- 238000007796 conventional method Methods 0.000 description 1
- 230000036461 convulsion Effects 0.000 description 1
- 150000001879 copper Chemical class 0.000 description 1
- 239000006071 cream Substances 0.000 description 1
- 230000001419 dependent effect Effects 0.000 description 1
- 230000036576 dermal application Effects 0.000 description 1
- 231100000223 dermal penetration Toxicity 0.000 description 1
- 238000000586 desensitisation Methods 0.000 description 1
- 238000011161 development Methods 0.000 description 1
- 230000018109 developmental process Effects 0.000 description 1
- 206010012601 diabetes mellitus Diseases 0.000 description 1
- 230000004069 differentiation Effects 0.000 description 1
- 238000002845 discoloration Methods 0.000 description 1
- 239000002552 dosage form Substances 0.000 description 1
- 239000003937 drug carrier Substances 0.000 description 1
- 235000013399 edible fruits Nutrition 0.000 description 1
- 235000014103 egg white Nutrition 0.000 description 1
- 210000000969 egg white Anatomy 0.000 description 1
- 239000003792 electrolyte Substances 0.000 description 1
- 238000004520 electroporation Methods 0.000 description 1
- 230000008030 elimination Effects 0.000 description 1
- 238000003379 elimination reaction Methods 0.000 description 1
- 230000002121 endocytic effect Effects 0.000 description 1
- 238000012407 engineering method Methods 0.000 description 1
- 238000003912 environmental pollution Methods 0.000 description 1
- 210000000981 epithelium Anatomy 0.000 description 1
- 239000010642 eucalyptus oil Substances 0.000 description 1
- 229940044949 eucalyptus oil Drugs 0.000 description 1
- 230000029142 excretion Effects 0.000 description 1
- 229950003499 fibrin Drugs 0.000 description 1
- 239000012847 fine chemical Substances 0.000 description 1
- 238000010579 first pass effect Methods 0.000 description 1
- 150000002220 fluorenes Chemical class 0.000 description 1
- 230000004927 fusion Effects 0.000 description 1
- 210000004907 gland Anatomy 0.000 description 1
- 239000011521 glass Substances 0.000 description 1
- 235000013922 glutamic acid Nutrition 0.000 description 1
- 239000004220 glutamic acid Substances 0.000 description 1
- ZDXPYRJPNDTMRX-UHFFFAOYSA-N glutamine Natural products OC(=O)C(N)CCC(N)=O ZDXPYRJPNDTMRX-UHFFFAOYSA-N 0.000 description 1
- 125000005456 glyceride group Chemical group 0.000 description 1
- 230000002439 hemostatic effect Effects 0.000 description 1
- 229920000669 heparin Polymers 0.000 description 1
- 229960002897 heparin Drugs 0.000 description 1
- 229940088597 hormone Drugs 0.000 description 1
- 239000005556 hormone Substances 0.000 description 1
- 230000036571 hydration Effects 0.000 description 1
- 238000006703 hydration reaction Methods 0.000 description 1
- 150000002430 hydrocarbons Chemical group 0.000 description 1
- 239000001257 hydrogen Substances 0.000 description 1
- 229910052739 hydrogen Inorganic materials 0.000 description 1
- 230000003301 hydrolyzing effect Effects 0.000 description 1
- 206010020718 hyperplasia Diseases 0.000 description 1
- 230000002218 hypoglycaemic effect Effects 0.000 description 1
- 230000006698 induction Effects 0.000 description 1
- 238000007689 inspection Methods 0.000 description 1
- 229940125396 insulin Drugs 0.000 description 1
- 150000002500 ions Chemical class 0.000 description 1
- 230000007794 irritation Effects 0.000 description 1
- 230000003780 keratinization Effects 0.000 description 1
- 229940124280 l-arginine Drugs 0.000 description 1
- 150000002596 lactones Chemical class 0.000 description 1
- 238000002386 leaching Methods 0.000 description 1
- 238000004895 liquid chromatography mass spectrometry Methods 0.000 description 1
- 210000004185 liver Anatomy 0.000 description 1
- 230000033001 locomotion Effects 0.000 description 1
- 210000001365 lymphatic vessel Anatomy 0.000 description 1
- 238000001819 mass spectrum Methods 0.000 description 1
- 239000011159 matrix material Substances 0.000 description 1
- 230000001404 mediated effect Effects 0.000 description 1
- 239000001525 mentha piperita l. herb oil Substances 0.000 description 1
- 239000007769 metal material Substances 0.000 description 1
- 229930182817 methionine Natural products 0.000 description 1
- 125000001160 methoxycarbonyl group Chemical group [H]C([H])([H])OC(*)=O 0.000 description 1
- 244000000010 microbial pathogen Species 0.000 description 1
- 230000004089 microcirculation Effects 0.000 description 1
- 210000003632 microfilament Anatomy 0.000 description 1
- 230000005012 migration Effects 0.000 description 1
- 238000013508 migration Methods 0.000 description 1
- 230000000116 mitigating effect Effects 0.000 description 1
- 238000002156 mixing Methods 0.000 description 1
- 230000001333 moisturizer Effects 0.000 description 1
- 230000003020 moisturizing effect Effects 0.000 description 1
- 125000001419 myristoyl group Chemical group O=C([*])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])[H] 0.000 description 1
- 239000002105 nanoparticle Substances 0.000 description 1
- 229930014626 natural product Natural products 0.000 description 1
- 210000003739 neck Anatomy 0.000 description 1
- 230000002232 neuromuscular Effects 0.000 description 1
- 239000002858 neurotransmitter agent Substances 0.000 description 1
- 230000035764 nutrition Effects 0.000 description 1
- WWZKQHOCKIZLMA-UHFFFAOYSA-N octanoic acid Chemical compound CCCCCCCC(O)=O WWZKQHOCKIZLMA-UHFFFAOYSA-N 0.000 description 1
- 230000008520 organization Effects 0.000 description 1
- 230000000149 penetrating effect Effects 0.000 description 1
- 239000003961 penetration enhancing agent Substances 0.000 description 1
- 235000019477 peppermint oil Nutrition 0.000 description 1
- 230000008447 perception Effects 0.000 description 1
- 231100000435 percutaneous penetration Toxicity 0.000 description 1
- 230000002093 peripheral effect Effects 0.000 description 1
- 238000002823 phage display Methods 0.000 description 1
- 238000010587 phase diagram Methods 0.000 description 1
- 230000000704 physical effect Effects 0.000 description 1
- 239000002504 physiological saline solution Substances 0.000 description 1
- 239000000049 pigment Substances 0.000 description 1
- 230000003518 presynaptic effect Effects 0.000 description 1
- 230000035755 proliferation Effects 0.000 description 1
- 238000000746 purification Methods 0.000 description 1
- 239000010453 quartz Substances 0.000 description 1
- 238000004064 recycling Methods 0.000 description 1
- 230000001105 regulatory effect Effects 0.000 description 1
- 230000008439 repair process Effects 0.000 description 1
- 235000020195 rice milk Nutrition 0.000 description 1
- 238000007665 sagging Methods 0.000 description 1
- 231100000241 scar Toxicity 0.000 description 1
- 210000002374 sebum Anatomy 0.000 description 1
- 230000028327 secretion Effects 0.000 description 1
- 230000035945 sensitivity Effects 0.000 description 1
- 210000000697 sensory organ Anatomy 0.000 description 1
- 230000001568 sexual effect Effects 0.000 description 1
- 238000007493 shaping process Methods 0.000 description 1
- VYPSYNLAJGMNEJ-UHFFFAOYSA-N silicon dioxide Inorganic materials O=[Si]=O VYPSYNLAJGMNEJ-UHFFFAOYSA-N 0.000 description 1
- 150000003384 small molecules Chemical class 0.000 description 1
- 238000003746 solid phase reaction Methods 0.000 description 1
- 238000010671 solid-state reaction Methods 0.000 description 1
- 238000010183 spectrum analysis Methods 0.000 description 1
- 238000003860 storage Methods 0.000 description 1
- 239000013589 supplement Substances 0.000 description 1
- 230000001629 suppression Effects 0.000 description 1
- 238000004347 surface barrier Methods 0.000 description 1
- 230000005062 synaptic transmission Effects 0.000 description 1
- 230000008337 systemic blood flow Effects 0.000 description 1
- 230000001839 systemic circulation Effects 0.000 description 1
- 150000003505 terpenes Chemical class 0.000 description 1
- 235000007586 terpenes Nutrition 0.000 description 1
- 230000001225 therapeutic effect Effects 0.000 description 1
- 230000000699 topical effect Effects 0.000 description 1
- 238000013271 transdermal drug delivery Methods 0.000 description 1
- 230000017105 transposition Effects 0.000 description 1
- OUYCCCASQSFEME-UHFFFAOYSA-N tyrosine Natural products OC(=O)C(N)CC1=CC=C(O)C=C1 OUYCCCASQSFEME-UHFFFAOYSA-N 0.000 description 1
- 239000000341 volatile oil Substances 0.000 description 1
- 239000002699 waste material Substances 0.000 description 1
Classifications
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K8/00—Cosmetics or similar toiletry preparations
- A61K8/18—Cosmetics or similar toiletry preparations characterised by the composition
- A61K8/30—Cosmetics or similar toiletry preparations characterised by the composition containing organic compounds
- A61K8/64—Proteins; Peptides; Derivatives or degradation products thereof
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K8/00—Cosmetics or similar toiletry preparations
- A61K8/02—Cosmetics or similar toiletry preparations characterised by special physical form
- A61K8/04—Dispersions; Emulsions
- A61K8/06—Emulsions
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61Q—SPECIFIC USE OF COSMETICS OR SIMILAR TOILETRY PREPARATIONS
- A61Q19/00—Preparations for care of the skin
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K7/00—Peptides having 5 to 20 amino acids in a fully defined sequence; Derivatives thereof
- C07K7/04—Linear peptides containing only normal peptide links
- C07K7/08—Linear peptides containing only normal peptide links having 12 to 20 amino acids
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K2800/00—Properties of cosmetic compositions or active ingredients thereof or formulation aids used therein and process related aspects
- A61K2800/10—General cosmetic use
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K2800/00—Properties of cosmetic compositions or active ingredients thereof or formulation aids used therein and process related aspects
- A61K2800/40—Chemical, physico-chemical or functional or structural properties of particular ingredients
- A61K2800/41—Particular ingredients further characterized by their size
- A61K2800/413—Nanosized, i.e. having sizes below 100 nm
-
- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
- Y02P—CLIMATE CHANGE MITIGATION TECHNOLOGIES IN THE PRODUCTION OR PROCESSING OF GOODS
- Y02P20/00—Technologies relating to chemical industry
- Y02P20/50—Improvements relating to the production of bulk chemicals
- Y02P20/55—Design of synthesis routes, e.g. reducing the use of auxiliary or protecting groups
Landscapes
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- General Health & Medical Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Animal Behavior & Ethology (AREA)
- Public Health (AREA)
- Veterinary Medicine (AREA)
- Birds (AREA)
- Epidemiology (AREA)
- Organic Chemistry (AREA)
- Biochemistry (AREA)
- Biophysics (AREA)
- Genetics & Genomics (AREA)
- Medicinal Chemistry (AREA)
- Molecular Biology (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- Dispersion Chemistry (AREA)
- Dermatology (AREA)
- Cosmetics (AREA)
- Medicinal Preparation (AREA)
- Medicines That Contain Protein Lipid Enzymes And Other Medicines (AREA)
Abstract
The present invention relates to skin anti-aging and the fields such as beauty and skin care health care and daily cosmetics, be it is a kind of can Transdermal absorption cell-penetrating peptide-acetyl group Argireline nano-emulsion and preparation method thereof.It first designs and artificial synthesized cell-penetrating peptide-acetyl group Argireline prepares its nano-emulsion again.Invention uses nano-emulsion and cell-penetrating peptide modification technique synergistic effect to improve acetyl group Argireline transdermal penetration ability and Transdermal absorption effect; to reduce peptide degradation; peptide molecule stable structure and bioactivity are maintained, is improved using the peptide as the dissolubility and validity of cosmetic purpose drug or functional component.Cell-penetrating peptide modifies acetyl group Argireline nano-emulsion; it reduces and easily causes the S/C of skin irritatin and allergy that application security and comfort can be improved; and product is prepared and is carried out in room temperature; without heating and providing energy; it can mass production without special installation; not only it is conducive to industrialization, but also conducive to environmental protection and saves the energy.
Description
Technical field
The present invention relates to the related disciplines such as human body face beauty, skin-protection and health-care and its skin anti-aging cosmetics necks
Domain, and in particular to it is a kind of can be used for skin of face beauty and can the cell-penetrating peptide of transdermal penetration and Transdermal absorption carry acetyl group six
Win peptide (one acetyl group Argireline of cell-penetrating peptide) nano-emulsion and preparation method thereof.
Background technology
1, the background of related of skin barrier function and its Transdermal absorption
Human skin (Skin) is the maximum tissue organ for being covered in body surface, it, which not only has, protects entire body and have
There are the physiological functions such as secretion, excretion, perception and metabolism, and it is also the most important natural cover for defense of human external, can make body
Interior various tissues and organ from it is extraneous it is physical, chemically, the substances such as mechanicalness and pathogenic microorganism immerse body and invade
Evil body, while also preventing internal moisture, electrolyte and other material streams indecorous outer.In fact, what human skin had
These physiological functions (including skin barrier function) institutional framework special with it is closely related.Because human skin tissue by
It is divided into three layers, i.e. epidermis, skin corium and subcutaneous tissue outside to inside.Outermost epidermis microstructure can divide again from inside to outside
For basal cell layer, stratum spinosum epidermidis, stratum granulosum, hyaline layer and cuticula, (but skin of face tissue epidermis there is no hyaline layer knot
Structure).Human epidermal thickness is about 0.070-0.120mm, is mainly made of epithelial cells and dentritic cell.In epidermis, respectively
The transdermal penetration and Transdermal absorption of kind functional component are mainly related with the angling process and its degree of epithelial cells.In skin histology
The skin corium of interbed is then the structure organization between epidermis and subcutaneous tissue, with the collagenous fibres for belonging to connective tissue
It is reticular fibre to interweave jointly with elastic fibers.Under Electronic Speculum, skin corium is in fibrous reticular structure, collagenous fibres and elastic fibers
It is interweaved, is crisscross, centre is filled with unbodied matrix, and has cutaneous appendages and abundant nerve, blood vessel and leaching
Hand shaft etc..The undermost subcutaneous tissue of skin histology is located at then under corium, is a kind of containing the loose of a large amount of adipocyte
Tissue.There are sweat gland, blood vessel, lymphatic vessel nerve etc. in subcutaneous tissue.In general, existing in skin corium in skin histology abundant
Capillary, functional component as infiltration reach corium after can be absorbed quickly, and absorb transdermal functional component can quickly into
Enter blood circulation.And the outermost cuticula of epidermis is since keratinization degree is high, water content is low and by the extracellular base of lipid
Matter is wrapped up, and is the main barrier that various functional components enter in skin histology.Therefore, various functional component transdermal penetrations and thoroughly
One of the key factor that skin absorbs, is mainly how they penetrate the outermost cuticula of skin histology.
Theoretically, there are mainly two types of possible approach for external various functional component transdermal penetrations and Transdermal absorption： (1)
By epidermis approach, enter corium through cuticula and each layer of epidermis and body circulation be absorbed by capillary, this be effect at
Divide the main path of transdermal penetration and Transdermal absorption.In this approach, drug can pass through corneocyte to reach living Epidermis,
Living Epidermis can also be reached by corneocyte gap.Too due to corneocyte diffusional resistance, so functional component master
Cuticula is diffused through by iuntercellular.It is the multilayer lipid bilayer that lipoid molecule is formed, lipoid point between corneocyte
The hydrophilic segment bound water molecule of son forms aqueous area, and the hydrocarbon chain portion of lipoid molecule forms hydrophobic region, polar medicine molecule
Through the aqueous area infiltration between corneocyte, and non-polar drug molecule is permeated via hydrophobic region.(2) pass through hair follicle, sebaceous glands
With the approach such as sweat gland：I.e. functional component possibly through human skin tissue appendicle, such as hair follicle, sebaceous glands and sweat gland way
Diameter transdermal penetration and Transdermal absorption, to play corresponding biological action and physiologic function.
It acts on and saying from the effect of cosmetology function cosmetics, the performance and acquisition of cosmetology function, with wherein functional component
Transdermal penetration and Transdermal absorption have close association, and the mechanism of action in relation to transdermal penetration and Transdermal absorption includes mainly at present
Several hypothesis below：First, the integrality of keratoderma is directly destroyed by Physical effect so that functional component passes through skin
Breakage on skin be directly entered cuticula hereinafter, as using iontophoresis promote thoroughly, electroporation promote thoroughly, ultrasonic wave promote thoroughly, laser
Promote saturating, micropin and promotees saturating, thermal burn, mill skin etc.；Second, it is acted on by chemical method and makes class lipid bilayer in keratoderma
Ordered arrangement change, to a certain extent increase class lipid bilayer mobility, drug molecule be able to from cuticula dredge
Pine passes through in interval, such as dimethyl sulfoxide (DMSO), nitrogen ketone compounds, liposome, amino acid and its derivative, surface-active
Agent, dendritic macromole, volatile oil peppermint oil, eucalyptus oil, turpentine oil etc. and some natural products etc., in addition also terpenes
Class, alkaloids, lactone, alcohols etc. can also make functional component penetrate into skin；Third, by physically or chemically acting on
The binding ability for improving skin surface keratin and water, changes the aquation of keratoderma, and functional component is with hydration
Effect penetrates into skin；4th, sebaceous gland duct is penetrated by physically or chemically function influence skin accessory organ, such as transdermal enhancer
Interior dissolving sebum or lumen of gland wall epithelium lipid noble cells reduce the hydrophobicity in sebaceous gland duct, and ionic drug is promoted to penetrate
Skin.Transdermal enhancer expands sweat gland and hair follicle opening by cutin-softening layer, and one is gone forward side by side by skin to be conducive to functional component
Step absorbs.And currently used transdermal penetration and the correlation technique of Transdermal absorption are associated therewith.
Although percutaneous delivery administration or the administration of external preparation for skin approach encounter the technical bottleneck of the human skin natural cover for defense, it
Also represent a kind of purpose drug and the very valuable transport channel of functional component and penetration route.Percutaneous delivery administration or skin
Skin topical route administration it is simpler and convenient relative to other administering modes, also more safely and conveniently.Meanwhile this mode
Also less pain, also can avoid liver first-pass effect, and reduce risk of the injection needle recycling to transmission.Especially,
In medical cosmetology and cosmetic field, accelerate that the transdermal penetration of functional component and Transdermal absorption is promoted to be of great significance and apply
Value.
2, nano-emulsion and its background of related of application
The concept of nano-emulsion (nanoemulsion, NE) originates from last century the forties earliest, by Schulman and
Hoar etc. is put forward for the first time；Nineteen fifty-nine Schulman proposes nano-emulsion (NE) being officially named " micro emulsion again
The concept of this dispersion of (microemulsion, ME) ", the thus name one of nano-emulsion (NE) or referred to as micro emulsion (ME)
Directly use till today.However it is in fact really in medicine sector as the drug carrier system application of transdermal routes of administration
It took the lead in finding by scholars such as doctors Aitwood in 1974, research, application and open report.The nineties in last century, NE or ME make
The hot spot of medicine and Pharmaceutical study is had become for transdermal drug delivery system, after 2000, NE or ME become as now in the world
Medicine sector has the research field of huge applications potentiality, just starts to lead in skin-protection and health-care and effect cosmetics from 2006
Domain formally occurs.
Though its science Forming Mechanism of NE or ME and transdermal effect mechanism are intricate, their foundational system composition and structure
At ingredient mainly by pseudo-ternaries phases such as water phase, oil phase, emulsifier/assistant for emulsifying agent or surfactants/cosurfactants
Liquid collectively constitutes, can be in room temperature by this pseudo-ternary phase liquid according to the determining proper proportion relationship of scientific design and experiment
Under the conditions of prepared using pertinent instruments and method and form thermodynamically stable colloidal dispersion system.NE or ME under normal conditions
The clear appearance of foundational system is transparent, micro-strip opalescence and good dispersion, mobility are good, liquid viscosity is low, isotropism, grain
Diameter is evenly distributed, diameter range is between 10~110nm.NE or ME is to there is no a kind of NE or ME Forming Mechanisms energy complete so far
Scientific theory that is whole and definitely illustrating and explain NE or ME System formings, such as NE or ME systems negative interfacial tension form machine
It is all difficult unusual science, complete and accurately explain and explain that system, hybrid films, which form theoretical, geometry arrangement theory and R ratio theories etc.,
It is bright it, the negative interfacial tension Forming Mechanism for generally believing and accepting with ambit now is it is now recognized that more science and conjunction
The theory of reason and its explanation.This negative interfacial tension forms theoretical core and its key point, be NE or ME it is spontaneous and from
Dominant shape at dynamic process among, interfacial tension mechanism plays a crucial role, i.e. emulsifier or surfactant energy
The decline for enough causing oil-in-water (O/W) interfacial tension, appropriate assistant for emulsifying agent or cosurfactant is added can promote O/W circle again
Face tension continues to decline, until it is negative value i.e. negative interfacial tension to reach interfacial tension.This negative interfacial tension promotes system spontaneous
Interface is expanded, is adsorbed on interface to be continuously increased emulsifier or surfactant and assistant for emulsifying agent or cosurfactant, directly
It is gradually balanced to interfacial tension and reaches interfacial tension value and return back to zero.The negative interfacial tension shape occurred just because of above-mentioned moment
At to the spontaneous expansion in induction system interface and be autonomously formed NE or ME.And when NE or ME gradually form coalescence, its boundary
Face area also gradually reduces, and will form new negative interfacial tension in turn, and this new negative interfacial tension promotes interface again
Tension value returns back to zero, to the aggregation of contend with NE or ME, to maintain and support the stability of NE or ME.
As NE or ME Forming Mechanisms and its continuous of academic theory illustrate, they have been considered as presently the most excellent
Transdermal drug or functional component transmit one of carrier.They not only have many application characteristics and Action advantage, can improve not
With the solubility of attribute effect drug ingedient, degrees of fusion and bearing capacity, but also the NE that can be formed by its pseudo-ternary phase liquid
Or ME systems are to useful effects such as horny layer of epidermis, stratum granulosum, stratum spinosum epidermidis and its cutaneous appendages, and then promote, increase and
The transdermal penetration and Transdermal absorption for accelerating different attribute effect drug ingedient, to play the maximum biology of effect drug ingedient
Effect.The high-performance bio effect of functional component is closely related with its bioavilability and its transdermal penetration and Transdermal absorption amount.
Therefore, using NE or ME as efficient percutaneous dosing transmit carrier, be not only widely used at present many new medicinal preparations it
In, and also begin to apply in some high-grade beauty and skin care effect cosmetics.Including skin-whitening, nti-freckle, reparation, it is crease-resistant,
Nutrition, moisturizing, sun-proof, desensitization, antiallergic, anti-aging, anti-oxidant, analgesia anesthesia, hemostatic nanoemulsion etc., make effect nanometer emulsion in skin
It is applied in skin nursing, beauty and health care and medical cosmetology and minimally invasive beauty and shaping and a new peak and fashion trend occurs.
3, cell-penetrating peptide and its background of related of application
Cell-penetrating peptide (cell-penetrating peptide, CPP) is at present not yet in beautifying skin cosmetic applications
Can percutaneous dosing transmit carrier, it is that one kind can determine that and carry destination protein or polypeptide drug and other functional ingredient in field of medicaments
And its transdermal small peptide can be promoted, this small peptide has the function of carrying function and percutaneous delivery, is current carrying polypeptide
One of class functional component transdermal penetration and the optimal percutaneous delivery carrier of Transdermal absorption.It has not yet to see to skin have it is any
Damage or stimulation and allergic reaction, itself also without individual pharmacological activity and effect, and it have metastable molecular structure and
Physicochemical property, and have with the good connection of its entrained destination protein or polypeptide drug and other functional ingredient and it is preferable match and
Compatibility；It relatively waits so long feature and advantage to the target area very fast, effect that works, and is that destination protein or polypeptide drug etc. pass through skin
Skin surface approach enters another simple, convenient and effective one of the transdermal penetration and Transdermal absorption approach of systemic circulation, and
To convenient administration mode and transdermal route another kind useful supplement, future is expected in some high-new, high-grade, efficient functional skins
The preparations such as anti-aging, skin-protection and health-care and cosmetics are researched and developed and are played a role in applying, and have wide development prospect
With can the phase economic results in society.
The biological function of cell-penetrating peptide or transdermal peptide, native conformation, molecular structure and amino acid composition are substantially similar.There is text
Offer report, cell-penetrating peptide can by carry large biological molecule purpose drug (albumen, polypeptide, nucleic acid and bigger particulate polymers
Deng) be directed through cell membrane and enter in different type cell, or even certain large biological molecule substances can be carried directly through blood
Brain barrier.It not only can actively or passively be absorbed at 37 DEG C by living cells, and can be 4 is found to cell-penetrating peptide study on mechanism
DEG C when enter in living cells, it is this wear film mechanism be considered it be a non-receptor, non-energy rely on and non-endocytosis dynamics
Mechanism.In recent years, scholar proposes a kind of new to wear film mechanism, it is believed that it is in being mediated based on clathrin that it, which enters born of the same parents' effect,
Gulp down effect, clathrin package indent film is allowed to that cytoplasm is detached from and be directly entered from cell membrane, this process may with it is thin
The negatively charged glycosaminoglycan of cellular surface occurs positive and negative charge with the cell-penetrating peptide with positive charge and attracts each other and act on, and
The positively charged amino acid residue in this cell-penetrating peptide surface and the negatively charged proteoglycans of cell membrane surface or aminoglucan
(such as Heparan sulfate, heparin) directly electrostatic interaction is that it wears film effect and by necessary key factor in cellular uptake
One of.Related cell-penetrating peptide wear film mechanism for explain cell-penetrating peptide binding molecule amount be more than 30,000 dalton purpose drug across
Film phenomenon is considered as a kind of Lipid Rafts cell endocytic effect that non-specific big vesica is formed, and this endocytosis by
Actin filament causes and mediates.And the poly arginine in the cell-penetrating peptide rich in arginine residues can be straight with small molecule purpose drug
Binding is closed, and by its electrostatic interaction or hydrogen bond joint efficiency promote it to penetrate after birth enter it is intracellular, when poly arginine with
Negatively charged cell surface can form ion pair after combining, and transposition and enter cytoplasm across after birth under the influence of film potential, then
Its biological action will be played in the purpose drug release to cytoplasm of cell-penetrating peptide combination again.
Cell-penetrating peptide can carry high molecular weight protein or polypeptide drug and other functional ingredient penetrates the after birth with double lipid structures
Into in cell, and transdermal peptide can then carry same or like purpose drug or functional component permeate through epidermis layer enters corium
It is interior.Body biomembrane and cell membrane are lipid bilayer structure, though skin outermost layer lipid film is by aliphatic acid, cholesterol and nerve
Amide forms, but is actually also lipid and negatively charged, the mechanism of action of this similarly suitable cell-penetrating peptide of structure.Transdermal peptide
It is likely to create of short duration sexual openness for the macromoleculars such as these destination proteins or polypeptide functional component or polymer drug
Percutaneous delivery channel reaches systemic blood circulation.Once there is scholar that cyclosporin is covalently attached to the poly- smart ammonia of cell-penetrating peptide
Sour haptamer, the ability for finding its transdermal penetration and this drug of Transdermal absorption and the effect for controlling acute inflammatory reaction have aobvious
It writes and is promoted, and the transdermal penetration of the cyclosporin without cell-penetrating peptide modification and Transdermal absorption and the ability for inhibiting acute inflammatory reaction
Differ greatly.It is another have research, it was also found that by specific simulated albumin or polypeptide moiety with to be rich in arginic intracellular carrier peptides non-
After covalent bond, these intracellular carrier peptides similarly have enhancing transport under Non-covalent binding form and promote these macromoleculars
The transdermal penetration and Transdermal absorption ability of simulated albumin or polypeptide drugs.It is this kind of with penetration cell film and transdermal enhancing effect
Polypeptide, which is also known as protein transduction domain, penetration cell film and can also carry certain purpose drugs and enter among skin histology.
Transdermal peptide and cell-penetrating peptide (transduction domain) have a structural features, i.e., being rich in has positively charged arginine and lysine residual
Base, and can transport binding peptide, oligonucleotide and liposome etc. and pass through mammalian cell membrane.It is this special rich in smart ammonia
The structure of acid is also possible to generate directly through skin because of charge effect by the charged components in its arginine side group and cell membrane
The effect of plasma membrane, it forms a polarity instant channel, makes carrying functional component by instantaneously opening the immobilized artificial membrane of skin surface
Transdermal penetration and Transdermal absorption are to intradermal, to play the biological action and physiologic function of functional component.And cell-penetrating peptide etc.
Basic protein transduction domain then be considered as by cell pinocytosis mode together with covalent linkage polypeptide or protein drug into
Enter in mammalian cell, and the endocytosis form of this cell pinocytosis may be then dependent on cell membrane recess, on film receptor and
The realizations such as clathrin.Moreover, there is scholar that Phage display in vivo is applied to dermal penetration enhancer at present
Research, and have found the transdermal bioactive peptide of energy efficient transportation protein drug that one is made of 11 amino acid.It should
Small peptide, which is simply mixed in physiological saline and is applied on diabetic model rats skin with insulin, can generate good hypoglycemic
Effect.In addition, this small peptide can also help other multiple proteins or hormone to enter in skin histology through skin barrier.Preliminary machine
This small peptide of reason studies have shown that may act on skin histology, and combine rear temporary opening skin by its positive and negative charge
The lipid film and its respective channel (including space between cells and hair follicle approach etc.) of barrier carry high molecular weight protein or polypeptide drugs afterwards
Enter cycle through skin.It is another to there is scholar to find a natural pore-forming peptide that increase cutaneous permeability, with skin surface
The permeability and permeability of skin can be enhanced when activating agent synergistic application.Separately there is scholar to design manually to be closed with polypeptide solid-state reaction method
Molecular structure alteration is carried out at the peptide analogues, and for having transdermal peptide, that is, changes the group of amino acid residue in transdermal peptide
At being scanned with lysine or arginine, increase its cations quantity and change position, also can reach and improve transdermal penetration and thoroughly
The effect of skin assimilation effect.Researcher thinks as a result, can not or be difficult to originally percutaneous dosing high molecular weight protein and polypeptide and
Nucleic acid drug when being covalently attached to cell-penetrating peptide poly arginine haptamer, be possible to penetrate skin plasma membrane function and
Effect, and be possible to assist through covalent linkage those of seem transdermal penetration and transdermal absorption factor it is very low or it could be theoretically argued that
The albumen or polypeptide functional component and purpose drug for being difficult to transdermal penetration and Transdermal absorption may also be sent out into skin histology
Wave good biological action and clinical therapeutic efficacy.This theoretical foundation and experimental result, only percutaneous dosing does not provide
A kind of new and innovative ways, and to functional form cosmetics, particularly certain contain macromolecular bioactivity functional component (egg
White matter, polypeptide, enzyme, nucleic acid) etc. functional forms cosmetics research and development and application provide a new thought and research side
Method and novel product.
4, beauty victory peptide and its background of related of application
Peptides (peptide) some scholars are referred to as to win peptide, are broadly divided into polypeptide and oligopeptides, are by two or two
The above amino acid condensation and a kind of bioactive substance being connected with peptide chain or amido bond, they are the intermediate products of protein,
It is also segment of the molecular structure between amino acid and protein.Peptide matters are compared with other materials in organism, most
Big feature, which is them, has very high bioactivity and bio-diversity, therefore is also referred to as biologically active polypeptide.Peptides
Or victory peptide be by more than 20 kind natural amino acids by peptide chain in different alignments and morphosis, i.e., from two peptides to polypeptide
Complex linear and loop configuration formed, they are derived from the multi-functional compounds of protein intermediate product and native conformation,
It is also most basic and important one of the substance for constituting life, they with the mode of action of natural bioactive in physical exertion
Important biological effect, and various vital movements, metabolism process and physiological function etc. in vivo, are also nearly all by specific amino
The active peptides or protein of acid sequence composition are dominated and are regulated and controled.Moreover, type, content and the bioactivity of polypeptide or victory peptide
It is also closely related with human skin tissue structure and function, and play under certain condition highly important biological action and
Physiological function, as skin histology and its cell division, proliferation, differentiation, chemotactic, migration, aggregation, new vessels generation, microcirculation,
Pigment is formed and the synthesis of skin histology albumen and regulation and control etc..
In medical cosmetology and cosmetics and field of fine chemical, the oligopeptides being made of two amino acid is usually known as two victory
Peptide, and then it is referred to as palmitoyl tripeptide 3 by what 3 amino acid formed, and so on, four victory peptides, SymPeptide, six victory can be divided into
Peptide ..., etc..It is continued to develop in particular with peptide synthesis technology and the victory peptide that is formed of different number amino acid is to skin
Skin acts on and its function is constantly disclosed, and the victory peptide of various structures is made gradually to start for beautifying face and skin-protection and health-care and delay
Skin aging is among resisting age of skin, and especially some biological actions are unique, amino acid composing quantity is less, molecular structure
Simply, molecular weight is small, ten victory peptides victory peptide below, such as SymPeptide, Argireline, seven victory peptides, eight victory peptides, nine victory peptides and its derivative
The applications such as Matrixyl -3, acetyl group hexapeptide -3, acetyl group tetrapeptide -5, palmityl tripeptides -1, palmityl tetrapeptide -3 and carnosine
It constantly comes out in cosmetics, and fashionable for a time, honourable a burst of.
However, 20th century the eighties once had the concept that scholar proposed protein hydrolysising fragment or keratin debris, also
I.e. by high molecular weight protein metallic substance, by protease hydrolytic, either Fermentation Engineering method is hydrolyzed to polypeptide or oligopeptides.But
The product degraded after hydrolysis due to these macro-molecular proteins is complicated, derivative polypeptide is because molecular weight its type that differs is various.
Due to being difficult to select polypeptide fragment and sequence, then the classification of polypeptide and type are difficult to determine, actively selection and clear polypeptide name
Claim difficulty, the purification of single polypeptide more difficult.Therefore the protein breakdown products or derivative of these hydrolysis or fermentation, it is practical
The mixture of upper exactly various protein, polypeptide and amino acid cannot still be referred to as real beauty victory peptide sterling.Real meaning
Above beauty those of related to beautifying skin wins peptide, can trace back to 1973 earliest and be found by Loren doctors Pickart et al.
Copper peptide (be also referred to as copper win peptide), this copper peptide can not only reduce scar tissue generation, while can also stimulate damaged skin group
It knits and actively heals, and the U.S. learns and this copper victory peptide is also found can improve skin aging situation, it is applied to people by them
Body skin repair, crease-resistant and hair hyperplasia etc..In addition, there is some beauty to win peptide, include mainly：(1) class signal wins peptide：Palm
Acyl tripeptides -1, palmityl tripeptides -5, Matrixyl -3, myristoyl pentapeptide -11, palmityl hexapeptide, hexapeptide -9 etc.；(2) refreshing
Class is inhibited to win peptide through mediator：Dipeptide ophiotoxin, pentapeptide -3, acetyl group hexapeptide -3, acetyl group octapeptide -1 etc.；(3) carrying class victory
Peptide：Copper wins peptide etc.；(4) other crease-resistant classes win peptide：Palmityl dipeptides -5, hexapeptide -8 or hexapeptide -10 etc..Wherein most widely used,
Effect Argireline most outstanding, also referred to as acetyl group Argireline -3, trade name Argireline, the class that is otherwise known as meat poisoning
Bacillus toxin, the victory peptide that it is made of 6 amino acid, since it can slow down muscular contraction force, allow it is of flaccid muscles, reduce or suppression
The generation of dynamic wrinkle and elimination microgroove.
5, class botulinum toxin (Argireline) and its background of related of application
Argireline is also referred to as " Argireline element " or " class botulinum toxin " by medical cosmetology circle at present, artificial synthesized
Derivative is acetyl group Argireline -3, and English name is Acetyl hexapeptide-3, and trade name is Argireline, it
It is one of most widely used skin wrinkle resisting functional component among current beauty victory peptide.Argireline is a kind of chemically synthesized skin
The crease-resistant functional component of external application, skin wrinkle resisting mechanism and the crease-resistant used botulinum toxin type A of injection are essentially identical, i.e. Argireline
The relaxant of similar skin of face muscle, energy selectively acting is last slightly in peripheral cholinergic nerve (kinesitherapy nerve), in Neuromuscular
(effect point of maximum intensity) inhibits presynaptic membrane to discharge neurotransmitter acetylcholine, the biography of effective block nerves medium at meat contact cynapse
Pass, and property of flaccid muscles caused to be benumbed, prevent muscle fibre from shrinking, muscle tone declines, the twin alleviation of flesh convulsion, facial expression muscle without
Method plays a role, and to make facial muscle loosen, reduces or eliminates the wrinkle of facial expression myokinesis formation, reaches flat and comforts dynamic
The purpose of line, static line and microgroove.Meanwhile class botulinum toxin (Argireline) can also effectively prevent catecholamine and excessively discharge to be produced
Raw wrinkle reduces facial muscles and repeats to affect to be formed by wrinkle, to effectively releive and inhibit forehead wrinkles on one's forehead, fish tail
The contraction and activity of line, glabella line, nose band, bridge of the nose line and wrinkle periphery muscle, help loosening all muscles, make elasticity of skin tissue
Submissive smooth lines are replied, face's microgroove is further reduced or eliminates.With age, the collagenous fibres of aging skin become
Directly, fibrin beam and skin table and it is all the more parallel, skin stretching, extension leeway is greatly reduced, and elasticity lowers, skin become sagging without
Power and there is wrinkle.The Argireline that Lipotec companies research and develop earliest obtained international monopoly early in 2002, and confirmed this
Kind of Argireline can local block nerves transmit contraction of muscle information, influence leather bag nerve conduction, so that facial muscle is loosened with
Smooth dynamic line, static line and microgroove.Lipotec companies are it is even contemplated that the effect after Argireline applications can be with A type meat poisonings
Toxin matches in excellence or beauty, but avoids the pain and the high disadvantage of use cost of injection botulinum toxin type A.Almost at the same time,
It is also the current most widely used crease-resistant victory peptide of beauty --- -3 (Palmitoyl of palmityl SymPeptide that Sederma companies, which have also produced,
Pentapeptide-3), but the crease-resistant mechanism of action of beauty victory peptide and Argireline is different, it is by local stimulation
With promote I types and type III collagen and fiber laminins generation, to increase skin thickness and reduce microgroove.Second
So having the function of the facial dynamic wrinkle of confrontation, being primarily due to it can block acyl group Argireline (Argireline) in face
Neurotransmission contraction of muscle information, influences leather bag nerve conduction, facial muscle is made to loosen, to smooth dynamic line, static line and thin
Line, or even there is scholar to think, its percutaneous external application can avoid the risk that previous botulinum toxin must be injected and use cost is higher
Insufficient and defect.Therefore, acetyl group Argireline (Argireline) is one of most widely used product in beauty victory peptide, and
It is considered skin anti-aging and the face of most application potential and foreground and vitality by cosmetics industry always so far
One of portion's anti-wrinkle cosmetic.Argireline used at present be mostly by the victory peptide of 6 Amino acid synthesis, can be with fluorenes when artificial synthesized
Protecting group of the methoxycarbonyl group (Fmoc) as a amino can apply alkali by it to the stability of acid and the characteristics of easily acted on alkali
Property compound makees deprotection agent.Therefore use protecting groups of the Fmoc as amino acid, by the carboxyl of first amino of Argireline with
The form of covalent bond is connected with solid carrier amino resins (Rjnk Amide MBHA Resin), to form product Fmoc-Arg
(pbf)-Rink Amide MBHA Resin be Peptide systhesis starting point, make on vector resin the polypeptide to be synthesized carry out by
A extension is condensed peptide reaction, final to synthesize ER-6-NH2 peptide chains, and the crude product that target product can be obtained in resin is sloughed in its acidolysis,
Then identification is carried out to crude product again and final products can be obtained in purifying.
6, the relevant technologies presently, there are defect or deficiency：
The optimum therapeuticing effect of external preparation for skin drug and the best beauty functions of functional form cosmetics with wherein add
Purpose drug and functional component are closely related, and the purpose drug of these additions and the key node of functional component only have maximum limit
Degree transdermal penetration and Transdermal absorption are possible to play best biological action and application effect.Make on domestic and international market at present
Gelling agent for external preparation for skin drug and cosmetics purposes is not only commonplace, but also type is various, too numerous to mention.Though
So, these gelling agents have the characteristics that certain and advantage compared with traditional emulsion and cream agent, are also fully consistent with outside human skin
With the quality standard smeared with face, and it is also fully consistent with the good appearance and physicochemical property of general gelling agent, and apply outside
Stretching and comfort level are also all fine, but these common gelling agents, and not only its grain size is larger, nor has promotion wherein
The characteristics of functional component and percutaneous penetration of drugs and Transdermal absorption, therefore, when practical application are to be extremely difficult to expection preferably to control
Treatment effect and beauty functions.
In terms of nano-emulsion is used as it is now recognized that good percutaneous dosing transmits one of carrier, theoretical research and application study
Achievement constantly come out, some have even achieved phasic results and breakthrough, especially nano-emulsion decentralized system
Grain size ultra micro and have reached nano scale level and Particle Distribution it is uniform, it is well dispersed, solubilized and contain effect it is all good, especially
It is the features such as it accelerates to promote wherein purpose drug and functional component transdermal penetration and preferable transdermal effect and advantage, receives people
Pay close attention to and pay attention to, like and favor.However, presently the most classical most of nano-emulsion foundational system be by oil phase, water phase,
The pseudo-ternaries phase liquid such as surfactants/cosurfactants forms, and the nano-emulsion system of formation is applied in external preparation for skin and face
Apparent greasy feeling, slightly irritation and tingling sensation, unhappy peculiar smell, smearing after smearing owe comfortable and preparation storage ability
The deficiencies of not good enough or defect protrude, in particular, in order to reach and maintain interfacial tension constantly to increase for negative value i.e. negative interfacial tension
Add emulsifier or surfactant and assistant for emulsifying agent or cosurfactant to be adsorbed on interface, makes in classical nano-emulsion foundational system
Contain higher concentration surfactants/cosurfactants, this nanoemulsions relative sensitivity skin irritation, especially to all
Such as destructiveness of protide or polypeptide functional component, it has also become nano-emulsion is in preparation for external application to skin and cosmetics field
The technical bottleneck and major obstacle of application.Moreover, the transdermal penetration and Transdermal absorption effect of nano-emulsion are also by factors shadow
It rings and interferes, including its constituent, composition, functional component type and drugloading rate and osmotic concentration gradient etc., this is also
Nano-emulsion preparation needs the problems and problem that solve and overcome at present.
Functional component work(is that some are extracted from animals and plants and marine organisms mesophytization mostly in Traditional skin anti-wrinkle cosmetic
Nutritional agents and moisturizer etc., and the addition of the victory peptide of the skin wrinkle resisting with certain bioactivity (Argireline), be skin wrinkle resisting
Biological cosmetic products provide completely new R&D direction and thinking.Argireline has been used as botulinum toxin substitute to be used for top grade at present
In cosmetics and beautifying skin and anti-wrinkle product, though it is that effect is more in current beauty victory peptide as Argireline element
Certainly and the beauty most widely used, molecular weight is smaller wins one of peptide, if percutaneous dosing reaches respective action position really,
It can inhibit skin catecholamine and acetylcholine by inhibiting the synthesis of SNARE acceptors after a certain period of time applied to skin of face
It excessively discharges and plays local block nerves and transmit contraction of muscle, influence leather bag nerve conduction, facial muscles is made to relax, to press down
The effects that making or mitigating dynamic and static wrinkle and facial lines.But there are still some defects and problems in its application, such as its conduct
Though water-soluble polypeptide molecular weight very little, but still hardly possible plays its due biological effect through skin, even if can be promoted using some
Permeation enhancers into its Transdermal absorption are assisted, and theoretic desired effect is still unable to reach, and clinical manifestation is crease-resistant reacts more
Slowly, curative effect is not notable.Moreover, the affiliated peptide class formation of Argireline makes its stability in conventional water-soluble medium or aqua liquid
It is poor, the degradable destruction in aqua of functional group and activated centre.It is added to current conventional formulation (such as aqua, essence
Magnificent liquid, emulsion, creme, paste, gelling agent etc.) it applies in skin surface or carries out facial smearing, it is difficult to transdermal penetration and thoroughly
The problems such as skin absorbs, in particular, the target area of Argireline required effect in dermal application is deeper, transdermal penetration and transdermal
The effect of absorption must more preferable its due biological action of competence exertion and physiologic function.
Transdermal peptide or cell-penetrating peptide belong to peptide class formation, and the stability among water-soluble medium or liquid preparation is same
It is poor, it is also easy degradation among aqua or destroys；And simple transdermal peptide or cell-penetrating peptide itself has no special pharmacology
Activity and curative effect effect；In addition, transdermal peptide or cell-penetrating peptide must connect or carry certain efficient purpose drugs and functional component
And the corresponding biological effect of matching competence exertion and physiologic function.In particular, as beautifying skin cosmetics
The carrier of middle functional component connection its connection or carries after beautifying skin functional component in body using not yet reported
Also there is not been reported at present for the experimental studies such as the actual effect of outer transdermal penetration and Transdermal absorption.
For this purpose, the present invention, which designs one kind, can not only reduce above application technical deficiency or defect, but also it is suitble to modern beautifying skin
Using, and also more preferably can permeate the preparation with Transdermal absorption by cutaneous routes, i.e., a kind of cell-penetrating peptide carries acetyl group Argireline and (wears film
Peptide-Argireline) nano-emulsion and preparation method thereof.
Invention content
The purpose of the present invention is made up primarily directed to many insufficient or defects present in background technology, is improved, is carried
High and innovation.The particular content of the present invention is to provide one kind not only can transdermal penetration and Transdermal absorption and molecular structure and biology
A kind of cell-penetrating peptide that activity is more stablized carries acetyl group Argireline (cell-penetrating peptide-acetyl group Argireline) nano-emulsion and its preparation side
Method.To achieve the goals above, the specific design and technical solution that the present invention uses are as follows：
1, present invention design
The technology of the present invention is divided into two parts：First, it completes cell-penetrating peptide and carries acetyl group Argireline (cell-penetrating peptide-acetyl group six
Win peptide) molecular structure and artificial synthesis design and synthesis；Second, it completes cell-penetrating peptide carrying acetyl group Argireline and (wears film
Peptide-acetyl group Argireline) nanometer dairy milk starting material forms and its design and preparation of ratio and preparation method.
2, the technology of the present invention
Mentality of designing according to the invention, a kind of cell-penetrating peptide-acetyl group Argireline nano-emulsion of the invention and its preparation side
The technical solution and specific implementation mode that method uses are divided into two parts.
First part is that the cell-penetrating peptide of the present invention carries acetyl group Argireline (cell-penetrating peptide-acetyl group Argireline) molecular structure
With artificial synthesis and analyze and identify：
The present invention uses the amino acid molecular structure of the artificial synthesized cell-penetrating peptide-acetyl group Argireline of chemical solid phase synthetic method
Sequence is Ac-EEMQRRYGRKKRRQRRR.Its artificial chemistry synthetic route and technological process are：Selection resin carrier is dichloro
Resin, the active site on resin are halogens chlorine, and Solid-phase synthesis peptides are firstly the need of resin swelling, then by first amino
The C-terminal carboxyl of acid is reacted with the active site chlorine on resin, after first amino acid is connected on resin, is carried out dehydrating condensation and is connect
Second amino acid takes off Fmoc protections again after the completion of condensation.According to the amino acid sequence repetitive operation of design, remaining has been connect successively
Amino acid and complete the acetylation of N-terminal, polypeptide is cut down with cutting reagent from resin finally, forms exposed carboxylic
Base.Its specific preparation method is as follows：(1) swelling of resin：Weighing dichloro resin 1.5g, (synthesis purpose peptide is 0.08mmol, institute
Degree of substitution with dichloro resin is 0.4mmol/g, and according to 7.5 times of excess, then required dichloro resin quality is 0.08/0.4*
7.5=1.5g), it is put into reaction column, 20 milliliters of DCM is then added in reaction column, vibrate 30min, activation is for use.(2) even
Adaper amino acid：DCM solvents are leached out by husky core, Fmoc-L-Arg (Pbf)-OH of 1.05 times of resin moles is added,
The DIEA for adding 10 times of resin moles is eventually adding a small amount of DMF dissolvings, vibrates 1h.It is handed over DMF and DCM after having reacted
For cleaning 6 times.(3) it is deprotected：20 milliliters of 20% piperidines/DMF solutions are added, are taken out after 5min.Add 20 milliliter of 20% piperazine
15min is vibrated in pyridine/DMF solution.(4) analysis detection：Piperidine solution is taken out, more than ten grainy resins are taken, is washed three times, is added with ethyl alcohol
Ninhydrin, pyridine, each drop of phenol, 105 DEG C of -110 DEG C of heating 5min, change navy blue, can be after under continued access one into positive reaction
A amino acid is feminine gender if non-discolouring, needs to be deprotected again.(5) it cleans for the first time：Successively with 15 milliliters of DMF, 15 milliliters
Methanol, 15 milliliters of DMF are respectively washed twice.(6) condensation reaction：The Fmoc-L-Arg (Pbf)-of 3 times of resin moles is added
OH, the HBTU of 3 times of resin moles are dissolved with a small amount of DMF, are added immediately the DIEA of 10 times of resin moles, are reacted
30min.(7) it cleans for second：Successively with 15 milliliters of DMF, 15 ml methanols, 15 milliliters of DMF are respectively washed twice.(8) peptide chain
Extension：Repetitive operation by the above process has connect remaining amino acid successively.
Table 1, it is artificial synthesized used in the process of the amino acid situation that arrives
(9) condensation of acetylation (AC) reaction and peptide：Acetylation is cell-penetrating peptide-the last one part of acetyl Argireline,
The acetic anhydride of 3 times of resin moles is added in reaction column, the pyridine of 3 times of resin moles reacts 30min.It is whole after AC is connected
The synthesis of peptide is completed with regard to reaction.Into the last contraction phase, since AC does not have Fmoc protections, so be just not necessarily to herein
It is deprotected.The contraction of peptide：With DMF washing reactions 3 times, DCM washing reactions 3 times, methanol washing reaction 3 times finally drains peptide
Resin.(10) amino acid side chain deprotection is cut with resin：The amino acid sequence of cell-penetrating peptide-acetyl Argireline is Ac-
EEMQRRYGRKKRRQRRR, wherein the Side chain protective group contained has：Pbf, Trt, Boc, Tbu, these types of protecting group is in acid item
It is unstable under part, and that cutting resin is TFA, so deprotection and cutting resin can be carried out at the same time.Configuration cuts liquid
The volume ratio of 15ml, wherein each component is：TFA (94.5%), water (2%), EDT (2.5%), TIS (1%).Resin is packed into
In flask, (30 DEG C) oscillation 2h of constant temperature.Lysate is dried up as possible with nitrogen, is subsequently poured into centrifuge tube, is poured slowly into ether.
It seals and is put into centrifuge and centrifuge 5min, outwell supernatant, lower section is white solid.It is washed 6 times with ether again, then room temperature is waved
It does to get crude product peptide.(11) purifying of efficient liquid phase (HPLC)：1. dissolving：Crude product peptide is put into vessel, it is dense with 30-50ml
Degree is completely dissolved for 50% acetonitrile solution, can slightly ultrasound 2min.2. filtering：With 0.45 μm of membrane filtration lysate.
3. analyzing：3 μ l solution are taken to analyze crude product with analysis level HPLC in order to subsequently prepare.Mobile phase is water and acetonitrile, time
30min, gradient elution, HPLC start gradients are first balanced 5min, and then sample introduction, start gradient are：Water 95%, acetonitrile 5%,
Terminating gradient is：Water 5%, acetonitrile 95%.4. preparing：The sample dissolved is done into sample introduction preparation and purifying.Prepare HPLC balances
10min, start gradient are：Water 95%, acetonitrile 5%, terminating gradient is：Water 25%, acetonitrile 75%, gradient timetable 40min.It collects
The sample come out from detector.5. identifying：The sample collected is sampled and carries out purity and mass spectrographic identification and analysis.
(12) it analyzes and identifies：1. Mass Spectrometric Identification is analyzed：Mass Spectrometric Identification is carried out using ESI-MS, that is, measures the actual molecular weight of synthetic peptide,
And its practical values for molecular weight and its theoretical molecular weight 2431.8 are compared and analyzed, the actually measured molecular weight of mass spectral analysis
Molecular weight is completely the same in allowable error positive and negative 2 or both for error, you can proves to successfully synthesize cell-penetrating peptide-acetyl six
Win peptide.2. HPLC is analyzed：Using LC3000 type high performance liquid chromatographs, chromatographiccondition：C18, reverse phase, 4.6mm*150mm,
Gradient elution.Start gradient is 5%A+95%B, and end gradient is 30%A+70%B, and the time is 30min, flow velocity 1.0ml/
Min, ultraviolet detection wavelength are 214nm, and sample size is 10 μ l.Mobile phase A is 0.1% trifluoroacetic acid in 100% acetonitrile, flowing
Phase B is 0.1% trifluoroacetic acid in 100% water.It observes its sterling and analyzes analysis top and area maximum peak through HPLC, with inspection
Measure the purity that cell-penetrating peptide-acetyl group Argireline reaches.
Second part is a kind of raw material of cell-penetrating peptide carrying acetyl group Argireline (cell-penetrating peptide-acetyl group Argireline) nano-emulsion
Composition ratio and preparation method and penetrating absorption：
1) a kind of cell-penetrating peptide-acetyl group Argireline nano-emulsion and preparation method thereof, it is characterised in that it is by following raw material
Composition and its weight percentage ranges are as follows：
Isopropyl myristate 7.32%-10.75%, Labraso 17.53%-20.12%, gather
Fatty acid glyceride 5.84%-6.71%, double distilled water 64.54%-68.97%, cell-penetrating peptide-acetyl group Argireline 0.01%-
0.2%.
2) each raw material that prepared by a kind of cell-penetrating peptide according to claim 1-acetyl group Argireline nano-emulsion is preferred
Optimum weight percentage is：
Isopropyl myristate 7.66%, Labraso 17.53%, polyglyceryl fatty acid ester
5.84%, double distilled water 68.87%, cell-penetrating peptide-acetyl group Argireline 0.1%.
A kind of preparation method that cell-penetrating peptide carries acetyl group Argireline (cell-penetrating peptide-acetyl group Argireline) nano-emulsion is as follows：
1) according to the proportioning of cell-penetrating peptide-acetyl group Argireline nanometer dairy milk starting material, using isopropyl myristate as oil phase (A)
It is spare；
2) proportioning for pressing cell-penetrating peptide-acetyl group Argireline nanometer dairy milk starting material makes acetyl Argireline, the mixing of appropriate distilled water
Be completely dissolved, then add distilled water be allowed to spare as water phase (C)；
3) proportioning for pressing cell-penetrating peptide-acetyl group Argireline nanometer dairy milk starting material, by Labraso, poly- sweet
Oil and fat acid esters is equal to 3: 1 ratio, is placed among another container, stirs, and is allowed to form S/C mixtures (B) standby
With；
4) again in oil phase (A): S/C mixtures (B): water phase (C) is equal to 7.66: 23.37: 68.97 ratio, takes respectively
S/C mixtures (B), water phase (C) are added in oil phase (A) container according to this；
5) time constant-temperature magnetic stirring apparatus at ambient temperature, is selected, and with 200rpmmin-1Rotating speed magnetic agitation
30min, you can obtain this cell-penetrating peptide-acetyl group Argireline nano-emulsion.
6) testing result for passing through Malvern particle size analyzer is shown：Malvern particle size determination result shows six victory of acetyl group
The average grain diameter of peptide nano-emulsion is between 24.62-163.2nm.
7) FRANZ methods transdermal experiment researches show that：Cell-penetrating peptide-acetyl group six in cell-penetrating peptide-acetyl group Argireline nano-emulsion
The skin permeation rate for winning peptide is 0.066%-0.18%.
3, feature of present invention
(1) we have substantially similar physicochemical property and biological characteristics according to medical or medicinal cell-penetrating peptide or transdermal peptide
Point, i.e. its stable in physicochemical property wear the Biological characteristics such as film or transdermal effect be apparent, successfully have one kind and wearing skin and wear film effect
The all good cell-penetrating peptide of fruit is connect and artificial synthesized this with transdermal penetration and Transdermal absorption effect with acetyl group Argireline
Cell-penetrating peptide carries acetyl group Argireline (cell-penetrating peptide-acetyl group Argireline).
(2) we by this confirmations of many researchs both at home and abroad at present it is existing penetrate skin surface barrier plasma membrane and have across
The cell-penetrating peptide of the double lipid layer structures of cell membrane, which carries acetyl group Argireline (cell-penetrating peptide-acetyl Argireline) conduct, can be added to skin
Then it is confirmed again with current research and is generally recognized by the bioactive ingredients among cosmetics and effect desired polypeptides
It matches and uses for the nano-emulsion system inherently with one of efficient percutaneous dosing transmission system, and play cell-penetrating peptide simultaneously and receive
The double effect of rice milk makes scientifically and rationally to play mutual aid collaboration and synergy effect between them, general at present so as to make
The state of better transdermal penetration and Transdermal absorption is formed all over the generally acknowledged beauty Argireline with good beautifying skin anti-wrinkle effect
Gesture and function.
(3) in order to overcome most of polypeptides matters (including beauty win peptide and Argireline etc.) in aqueous solution or other liquid phases
In relative instability and its characteristics of be easy to degrade and lose its biological activity and efficacy effect, we are not only with warp
The acetyl group Argireline of acetylation substitutes naked Argireline, and it is connected or modified with cell-penetrating peptide, but also by cell-penetrating peptide-
Acetyl group Argireline is prepared into nano-emulsion dosage form, i.e. cell-penetrating peptide-acetyl group Argireline nano-emulsion, make beauty functions therein at
Divide acetyl group Argireline that can maintain its good biological activity and physiologic function and keep its good transdermal penetration
With Transdermal absorption function.Moreover, more simple aqua, cell-penetrating peptide-acetyl group Argireline is less susceptible among nano-emulsion preparation
Degradation, bioactivity and holding time and term of validity compared with its in simple aqua it is longer and more long.
(4) since this cell-penetrating peptide-acetyl group Argireline nano-emulsion is modern based on nano-emulsion foundational system and nano-emulsion
It is prepared by technology, therefore, the grain size of liquid preparation prepared by more conventional or conventional method is more small, distribution and dispersion evenly,
Its appearance and sense organ are more excellent, and external application is smeared more comfortable.
(5) due to this cell-penetrating peptide-acetyl group Argireline nano-emulsion, efficient percutaneous delivery carrier nano-emulsion base had both been used
Plinth system, functional component acetyl group Argireline combine or are connected to not only a kind of cell-penetrating peptide that can wear skin but also can wear film, because
This, it has better transdermal penetration and Transdermal absorption effect.
(6) since this cell-penetrating peptide-acetyl group Argireline nano-emulsion has better transdermal penetration and Transdermal absorption effect,
Therefore the beauty biological effectiveness of its functional component acetyl group Argireline more preferably can be played and be embodied.
4, innovative point of the present invention
(1) the designed cell-penetrating peptide being added in cosmetics with synthesis of the present invention connects and carries acetyl group six
Win peptide (cell-penetrating peptide-acetyl group Argireline) and as human skin beauty functions ingredient and desired polypeptides application, at present state
Inside and outside there is not been reported, also without similar product, and therefore, the present invention has novelty.
(2) present invention is designed connects simultaneously with what is added in the pseudo-ternary phase nano-emulsion foundational system prepared containing cell-penetrating peptide
The beautifying effect nanometer emulsion for carrying acetyl group Argireline (cell-penetrating peptide-acetyl group Argireline), keeps acetyl group Argireline activity more steady
Fixed, transdermal penetration and Transdermal absorption effect are more preferable, and there is not been reported, also without similar product both at home and abroad at present, therefore, present invention tool
There is novelty.
5, beneficial effects of the present invention
(1) beauty functions ingredient Argireline of the invention is purpose polypeptide, and molecular structure passes through acetylation modification Cheng Yi
Acyl group Argireline, answers in cosmetics using physicochemical property more stable, and biological action should be more preferable.
(2) cell-penetrating peptide is connect by the present invention with acetyl group Argireline so that the acetyl group Argireline of purpose functional component passes through
Skin permeates and Transdermal absorption effect can be improved, and cosmetic result when practical application can be promoted.
(3) acetyl group Argireline is added to previous conventional formulation (such as aqua, Essence, emulsion, creme, paste, gelling agent
Deng) septum reset external application, it is difficult to reach expected transdermal penetration and Transdermal absorption effect.And the present invention is by cell-penetrating peptide and acetyl group six
Win peptide connection, and it is matched and combined with efficient percutaneous delivery carrier nano-emulsion system, not only cell-penetrating peptide can be played and be worn
Bark effect, and nano-emulsion system can also play the rush effect of oozing of percutaneous delivery acetyl group Argireline, the two has complementary advantages, acts on phase
Ji can make the transdermal penetration of the Argireline of purpose drug and functional component and Transdermal absorption effect more preferable, when practical application
Cosmetic result ought to be more preferably.
(4) the basic nano-emulsion system in nano-emulsion of the present invention has been subjected to that pseudo-ternary phase diagram is preferred, the watr-proportion of use
Increase, oil phase ratio decreases, and is prepared with the blank nano-emulsion foundational system that this is new, quality is finer and smoother, point
Dissipate evenly, particle is more small, grain size is good up to nano scale level, nanoparticle distribution, improve the comfortable of its face coating
Property.
(5) nano-emulsion type of the invention is oil-in-water type, can make cell-penetrating peptide carry purpose drug and functional component it
The dissolubility of acetyl group Argireline, solubilising and inclusiveness effect are more preferable.
(6) among the nano-emulsion system prepared of the present invention, surfactants/cosurfactants in other words emulsifier/
The surfactants/cosurfactants used in the more previous nano-emulsion document (including patent document) of assistant for emulsifying agent additive capacity
Emulsifier/assistant for emulsifying agent additive capacity significantly reduces in other words, therefore the tingling sensation of its face smearing and skin irritation are apparent
Decline, safety in utilization significantly improves.
(7) nano-emulsion type of the invention is not only oil-in-water type, and the oil phase wherein in pseudo-ternary phase and surface work
The adding proportion of property agent/cosurfactant also significantly reduces, and therefore, the greasy feeling and peculiar smell sensillary base of previous nano-emulsion originally disappear
It removes, the satisfaction that face is smeared significantly improves.
(8) acetyl Argireline and cell-penetrating peptide belong to the bioactive ingredients of polypeptide class formation, in water-soluble medium or
Stability is poor in aqua, its functional group and activated centre are easy degradation or are destroyed in aqua.And the nano-emulsion of the present invention
Preparation is similar to microballoon, and the cell-penetrating peptide of such as protide or polypeptide functional component can be made to carry the second of purpose drug and functional component
Acyl group Argireline wraps up wherein, its stability is made to improve, and the destructiveness of physicochemical property is apparent to be lowered, therefore, what face was smeared
Biological action and effect ought to be promoted.
(9) constituent, composition, the functional component type of cell-penetrating peptide-acetyl group Argireline nano-emulsion of the invention
With drugloading rate and osmotic concentration gradient etc., it is more conducive to its transdermal penetration and Transdermal absorption, this also partly solves previous liquid
State preparation needs the problems and problem that solve and overcome.
(10) either cell-penetrating peptide or transdermal peptide, itself has no special beauty functions or drug action, this hair
It is bright that dexterously cell-penetrating peptide is connected or carries purpose drug and functional component-acetyl group Argireline with cosmetology function, make
It not only has transdermal function, but also has cosmetology function.
(11) in addition, the present invention also has the advantages that compared with existing State of cosmetics：1. applying skill of the present invention
The stability higher for the more other State of cosmetics of this nano-emulsion that art is prepared, and its extension of validity；2. matching using the present invention
Main composition Transdermal absorption effect in this nano-emulsion of system is more preferable, transdermal absorption factor higher；3. using the present invention
The safety of its application of this nano-emulsion that technology is prepared is more preferable；4. being in room using this nano-emulsion that the technology of the present invention is prepared
Temperature is lower to be prepared, therefore more preferable for the bioactivity and biological action of heat labile cell-penetrating peptide-acetyl group Argireline；5. applying
This nano-emulsion that the technology of the present invention is prepared it is organoleptic more preferable, actual use comfort is more preferably.6. matching using the technology of the present invention
The preparation ingredient of this nano-emulsion of system is simplified, preparation process is simple, practical operation is easy, it is efficiently quick to prepare, does not need spy
Different instrument and equipment does not need extraneous offer energy, need not heat, and free from environmental pollution.Resource consumption and waste are reduced,
Product cost is set to substantially reduce.
It can be seen that invention of the present invention not only overcomes and solves above-mentioned background technology related to the present invention in modern skin
The many insufficient or defects applied in skin beauty and skin nursing, moreover, through the invention technology can also obtain it is more beneficial
Effect or benefit.
Description of the drawings：
Fig. 1 is a kind of cell-penetrating peptide-artificial synthesized technological process of acetyl group Argireline and Technology Roadmap.
Fig. 2 is the schematic arrangement of cell-penetrating peptide-acetyl Argireline.
Fig. 3 is that the Mass Spectrometric Identification of cell-penetrating peptide-acetyl group Argireline analyzes Testing and appraisal collection of illustrative plates.
Fig. 4 is that the HPLC of cell-penetrating peptide-acetyl group Argireline analyzes test map.
Fig. 5 is technological process and Technology Roadmap prepared by cell-penetrating peptide-acetyl group Argireline nano-emulsion.
Fig. 6 is the particle size distribution measuring analysis chart of cell-penetrating peptide-acetyl group Argireline nano-emulsion.
Fig. 7 is the transdermal absorption factor of cell-penetrating peptide-acetyl group Argireline nano-emulsion.
Specific implementation mode is illustrated
One of implementation example：
Cell-penetrating peptide carries the preparation method of acetyl group Argireline (cell-penetrating peptide-acetyl group Argireline) in the present invention：
1 experimental section
1.1 primary raw materials and reagent
Fmoc-Arg (Pbf)-OH (arginine), Fmoc-Gln (Trt)-OH (glutamine), Fmoc-Lys (Boc)-OH
(lysine), Fmoc-Gly-OH (glycine), Fmoc-Tyr (Tbu)-OH (tyrosine), Fmoc-Met-OH (methionine),
Fmoc-Glu (Tbu)-OH (glutamic acid), Ac (acetic anhydrides: pyridine=1: 1), 2-Chlorotrityl Chloride Resin
(2 chlorine resin), DMF (n,N-Dimethylformamide), DCM (dichloromethane), acetonitrile, HBTU (benzotriazole-N, N, N ', N '-
Tetramethylurea hexafluorophosphoric acid ester), DIEA (n,N-diisopropylethylamine), TFA (trifluoroacetic acid), TIS (tri isopropyl silane),
EDT (1,2- dithioglycol), ether, piperidines, ethyl alcohol, ninhydrin, phenol, pyridine.
1.2 key instrument
TDL-50 (table-type low-speed large capacity centrifuge) (Changzhou Mei Xiang Instrument Ltd.), HY-2 (the multi-purpose oscillations of speed governing
Device) (Changzhou Lang Yue instrument manufacturings Co., Ltd), FD-1A-50 (vacuum freeze drier) (the general gloomy limited public affairs of instrument and equipment in Nanjing
Department), SHZ-D (III) type (vacuum pump using circulatory water) (Bang Xi instruments Science and Technology Ltd.), electronic balance (the sharp product precision instrument in Changzhou
Device Co., Ltd), Peptide synthesizer, industrial nitrogen, (Beijing Ke Ruihai scientific instrument are limited for LC3000 types high performance liquid chromatograph
Company), 6120 LC/MS. of Agilent
1.3 solid phase synthesis process and step
1.3.1 synthetic route technological process：The resin carrier that the present invention selects is 2 chlorine resins, the active site on resin
For halogens chlorine, Solid-phase synthesis peptides are firstly the need of resin swelling, then by the work on the C-terminal carboxyl of first amino acid and resin
Property the reaction of site chlorine carry out dehydrating condensation after first amino acid is connected on resin and connect second amino acid, after the completion of condensation
Fmoc protections are taken off again.According to the amino acid sequence repetitive operation of design, remaining amino acid has been connect successively and has completed the acetyl of N-terminal
Change, polypeptide is cut down with cutting reagent from resin finally, forms exposed carboxyl.
1.3.2 the swelling of resin：Weighing dichloro resin 1.5g, (synthesis purpose peptide is 0.08mmol, dichloro resin used
Degree of substitution is 0.4mmol/g, and by 7.5 times of excess, then required dichloro resin quality is 0.08/0.4*7.5=1.5g), it is put into
In reaction column, 20 milliliters of DCM are then added in reaction column, vibrate 30min, activation is for use.
1.3.3 it connects first amino acid and DCM solvents is leached out by husky core, 1.05 times of resin moles of addition
Fmoc-L-Arg (Pbf)-OH, adds the DIEA of 10 times of resin moles, is eventually adding a small amount of DMF dissolvings, vibrates 1h.
With DMF and DCM alternately cleaning 6 times after having reacted.
1.3.4 20 milliliters of 20% piperidines/DMF solutions are added in deprotection, are taken out after 5min.Add 20 milliliter of 20% piperazine
15min is vibrated in pyridine/DMF solution.
1.3.5 piperidine solution is taken out in detection, takes more than ten grainy resins, is washed three times with ethyl alcohol, ninhydrin, pyridine, benzene is added
Each drop of phenol, 105 DEG C of -110 DEG C of heating 5min, change navy blue, can be after the next amino acid of continued access, if not into positive reaction
Discoloration is then feminine gender, needs to be deprotected again.
1.3.6 it cleans for the first time successively with 15 milliliters of DMF, 15 ml methanols, 15 milliliters of DMF are respectively washed twice.
1.3.7 Fmoc-L-Arg (Pbf)-OH of 3 times of resin moles is added in condensation, 3 times of resin moles
HBTU is dissolved with a small amount of DMF, is added immediately the DIEA of 10 times of resin moles, reacts 30min.
1.3.8 it cleans for second：Successively with 15 milliliters of DMF, 15 ml methanols, 15 milliliters of DMF are respectively washed twice.
1.3.9 the extension of peptide chain has connect remaining amino acid successively according to above method repetitive operation.
Amino acid in table 2, the present invention used in synthesis cell-penetrating peptide-acetyl group Argireline
1.3.10 the reaction of AC and the contraction acetylation of peptide are to wear skin peptide-the last one part of Argireline, in reaction column
In be added 3 times of resin moles acetic anhydride, the pyridine of 3 times of resin moles, react 30min.After AC is connected, whole peptide
Synthesis is completed with regard to reaction.Into the last contraction phase, since AC does not have Fmoc protections, so herein just without deprotection
?.The contraction of peptide：With DMF washing reactions 3 times, DCM washing reactions 3 times, methanol washing reaction 3 times finally drains peptide resin.
1.3.11 the amino acid sequence for cutting through skin peptide-Argireline of amino acid side chain deprotection and resin is Ac-
EEMQRRYGRKKRRQRRR, wherein the Side chain protective group contained has：Pbf, Trt, Boc, Tbu, these types of protecting group is in acid item
It is unstable under part, and that cutting resin is TFA, so deprotection and cutting resin can be carried out at the same time.Configuration cuts liquid
The volume ratio of 15ml, wherein each component is：TFA (94.5%), water (2%), EDT (2.5%), TIS (1%).Resin is packed into
In flask, (30 DEG C) oscillation 2h of constant temperature.Lysate is dried up as possible with nitrogen, is subsequently poured into centrifuge tube, is poured slowly into ether.
It seals and is put into centrifuge and centrifuge 5min, outwell supernatant, lower section is white solid.It is washed 6 times with ether again, then room temperature is waved
It does to get crude product peptide.
1.3.12 HPLC purifies (1) dissolving and crude product peptide is put into vessel, with the acetonitrile of 30-50ml a concentration of 50%
Aqueous solution is completely dissolved, can slightly ultrasound 2min.(2) 0.45 μm of membrane filtration lysate is filtered.(3) analysis takes 3 μ l molten
Liquid is subsequently prepared with analysis level HPLC analysis crude products.Mobile phase is water and acetonitrile, time 30min, gradient elution, first will
HPLC start gradients balance 5min, and then sample introduction, start gradient are：Water 95%, acetonitrile 5%, terminating gradient is：Water 5%, second
Nitrile 95%.(4) it prepares and the sample dissolved is done into sample introduction preparation.It prepares HPLC and balances 10min, start gradient is：Water 95%, second
Nitrile 5%, terminating gradient is：Water 25%, acetonitrile 75%, gradient timetable 40min.Collect the sample come out from detector.
1.3.13 identify that the sample that will be collected is sampled and does purity and mass spectrographic identification.
2 analyze and identify
2.1 Mass Spectrometric Identifications are analyzed：Using ESI-MS to the actual molecular weight of cell-penetrating peptide of the present invention-acetyl group Argireline into
5 target peaks for carrying different charges are obtained by Mass Spectrometer Method analysis, respectively in row Mass Spectrometric Identification：
[M+3H]3H+Karyoplasmic ratio is 811.4, and actual measurement molecular weight is 811.4*3-3=2431.2,
[M+4H]4H+Karyoplasmic ratio is 608.9, and actual measurement molecular weight is 608.9*4-4=2431.6,
[M+5H]5H+Karyoplasmic ratio is 487.4, and actual measurement molecular weight is 487.4*5-5=2432.0,
[M+6H]6H+Karyoplasmic ratio is 406.3, and actual measurement molecular weight is 406.3*6-6=2431.8,
[M+7H]7H+Karyoplasmic ratio is 348.4, and actual measurement molecular weight is 348.4*7-7=2431.8.And cell-penetrating peptide-acetyl
The theoretical molecular weight of base Argireline is also just 2431.8.The usual actually measured molecular weight of mass spectrum and theoretical molecular weight allow to miss
Difference is ± 2.And both the theoretical molecular weight of cell-penetrating peptide of the present invention-acetyl group Argireline and actual molecular weight are consistent, thus can demonstrate,prove
Exactly cell-penetrating peptide-acetyl group Argireline that the bright present invention synthesizes.The Mass Spectrometric Identification of cell-penetrating peptide of the present invention-acetyl Argireline is analyzed
Testing and appraisal collection of illustrative plates (Fig. 3 for referring to description of the drawings).
2.1 efficient liquid phases (HPLC) analysis detection：The present invention uses LC3000 type high performance liquid chromatographs, chromatography item
Part：C18, reverse phase, 4.6mm*150mm, gradient elution.Start gradient is 5%A+95%B, and end gradient is 30%A+70%B,
Time is 30min, and flow velocity 1.0ml/min, ultraviolet detection wavelength is 214nm, and sample size is 10 μ l.Mobile phase A is 0.1%
For trifluoroacetic acid in 100% acetonitrile, Mobile phase B is 0.1% trifluoroacetic acid in 100% water, to cell-penetrating peptide-acetyl group Argireline into
Row HPLC analysis detections, testing result show：Cell-penetrating peptide of the present invention-acetyl group Argireline sterling is analyzed through HPLC and determines reservation
The top of time 8.219 is area maximum peak, is herein the accessible purity of the synthetic product, which passes through
Its integral can calculate its purity and have reached 98.76%.The HPLC of cell-penetrating peptide of the present invention-acetyl Argireline analyzes test map
(Fig. 3 for referring to description of the drawings).
It is embodied the two of citing：
A kind of cell-penetrating peptide-acetyl group Argireline nano-emulsion and preparation method thereof (for preparing 100g)：
1. primary raw material：
1. mainly preparing raw material：Isopropyl myristate (IPM), Labraso (Labrasol),
Polyglyceryl fatty acid ester (Plurol Oleique CC497), double distilled water；2. main composition：Cell-penetrating peptide-acetyl group six
Win peptide；
2. mainly preparing equipment
Ten a ten thousandth electronic balances, a ten thousandth electronic balance, one thousandth pharmacist balance, liquid flash mixer,
Time constant-temperature magnetic stirring apparatus, quartz ampoule glass aqua bi-distilling apparatus, considerable low-temperature centrifuge, sterilizing filter, various rule
Lattice container, flask, beaker.
3. main preparation process
1. accurately weighing the isopropyl myristate of 7.66g weight with one thousandth pharmacist balance, it is immediately placed on by washing
Among the 100mL cleaning conical flasks to disappear.It is used in combination marking pen to understand that label is (A)
2. weighing cell-penetrating peptide-acetyl group Argireline 100mg respectively with the analytical precision balances of ten a ten thousandths again to be placed in together
Among the conical flask of one 100mL cleaning by decontamination sterilizing.The double distilled water for being directly added into 40mL fills respectively to above-mentioned
Among the 100mL conical flasks of kind functional component, capping, and this conical flask is immediately placed on liquid flash mixer；It beats
Liquid flash mixer is opened, and is adjusted to II grades, is sufficiently mixed and uniformly is allowed to be completely dissolved, after 3-5min, closing liquid is fast
Fast mixer, removes conical flask, and it is (C) to be used in combination color mark pen clearly to mark.
3. according to designed Labraso in advance：17.53 grams of sad last of the ten Heavenly stems are accurately weighed respectively
Acid polyethylene glycol glyceride and 5.84 grams of polyglyceryl fatty acid esters, and the two is placed in the 100mL cleaning triangles by decontamination
Among flask；Capping, and this conical flask is immediately placed on liquid flash mixer, liquid flash mixer is opened, and will
It is adjusted to II grades, is sufficiently mixed uniformly, is allowed to form emulsifier/assistant for emulsifying agent (S/C) mixture, after 3-5 minutes, closing liquid
Flash mixer removes conical flask, and marking pen is used in combination to understand that label is mixture (B).
4. it is last, then in oil phase (A): S/C mixtures (B): water phase (C) is equal to 7.66: 23.37: 68.97 ratio, takes
Water phase (C) is directly added in the 100mL conical flasks in above-mentioned oil phase (A)；Then, then caprylic capric polyethylene glycol glycerol is taken
The S/C mixtures (B) that ester and polyglyceryl fatty acid ester are thoroughly mixed to form are directly added to above-mentioned oil phase (A) and water phase (C) is mixed
In the 100mL conical flasks of conjunction.
5. selecting time constant-temperature magnetic stirring apparatus under the conditions of 25 DEG C of room temperature immediately, with 200rpmmin-1Rotating speed magnetic force
Stir 30min.
6. timeing closing constant temperature blender with magnetic force removes 100mL conical flasks, it is observed that its appearance is limpid, transparent, stream
Body is good, good dispersion, is evident that opalescence, this cell-penetrating peptide of as 100g carry acetyl group Argireline nano-emulsion.
Embodiment of the present invention is only the description carried out to the preferred embodiment of the present invention, not to this hair
Bright conception and scope is defined, and under the premise of not departing from design philosophy of the present invention, engineers and technicians are to this in this field
The all variations and modifications that the technical solution of invention is made, should all fall into protection scope of the present invention, and the present invention is claimed
Technology contents have all been recorded in detail in the claims.
Claims (3)
1. a kind of cell-penetrating peptide-acetyl group Argireline nano-emulsion and preparation method thereof, it is characterised in that it is to be made of following raw material
And its weight percentage ranges are as follows：
Isopropyl myristate 7.32%-10.75%, Labraso 17.53%-20.12%, polyglycereol
Aliphatic ester 5.84%-6.71%, double distilled water 64.54%-68.97%, cell-penetrating peptide-acetyl group Argireline 0.01%-
0.2%.
2. each raw material prepared by a kind of cell-penetrating peptide according to claim 1-acetyl group Argireline nano-emulsion is preferably best
Weight percent is：
Isopropyl myristate 7.66%, Labraso 17.53%, polyglyceryl fatty acid ester 5.84%,
Double distilled water 68.87%, cell-penetrating peptide-acetyl group Argireline 0.1%.
3. a kind of preparation method of cell-penetrating peptide according to claim 1-acetyl group Argireline nano-emulsion, it is characterised in that it
Include the following steps：
First, the molecular structure and its sequence of design cell-penetrating peptide-acetyl group Argireline simultaneously carry out artificial synthesized：The present invention designs
And cell-penetrating peptide-acetyl group the Argireline synthesized is the small peptide containing 17 amino acid, amino acid composition and molecular structure sequence
For Ac-EEMQRRYGRKKRRQRRR, artificial synthesized method uses chemical solid phase synthetic method.Its specific artificial synthesis is：
(1) weigh dichloro resin 1.5g (synthesis purpose peptide is 0.08mmol, and the degree of substitution of dichloro resin used is 0.4mmol/g, according to
7.5 times of excess, then required dichloro resin quality is 0.08/0.4*7.5=1.5g), it is put into reaction column, then in reaction column
In 20 milliliters of DCM (dichloromethane) are added, vibrate 30min, swelling and the activation for completing dichloro resin are for use.(2) pass through husky core
DCM (dichloromethane) solvent is leached out, Fmoc-L-Arg (Pbf)-OH (arginine) of 1.05 times of resin moles is added, then add
The DIEA (n,N-diisopropylethylamine) for entering 10 times of resin moles, is eventually adding a small amount of DMF (n,N-Dimethylformamide)
1h is vibrated in dissolving.With DMF (n,N-Dimethylformamide) and DCM (dichloromethane) alternately cleaning 6 times after having reacted, complete first
The connection of a amino acid.(3) 20 milliliters of 20% piperidines/DMF solutions are added, are taken out after 5min.Add 20 milliliter of 20% piperidines/
DMF solution vibrates 15min, completes its deprotection reaction.(4) piperidine solution is taken out, more than ten grainy resins are taken, is washed three times with ethyl alcohol,
Ninhydrin is added, pyridine, each drop of phenol, 105 DEG C of -110 DEG C of heating 5min, for change navy blue into positive reaction, such as non-positive is anti-
It answers, then needs to re-start deprotection.(5) successively with 15 milliliters of DMF, 15 ml methanols, 15 milliliters of DMF are respectively washed twice.
(6) Fmoc-L-Arg (Pbf)-OH, HBTU of 3 times of resin moles of 3 times of resin moles is added, it is molten with a small amount of DMF
Solution is added immediately the DIEA of 10 times of resin moles, reacts 30min.(7) again successively with 15 milliliters of DMF, 15 ml methanols, 15
Milliliter DMF is respectively washed twice.(8) repeat to operate according to above method, i.e., according to cell-penetrating peptide of the present invention-acetyl Argireline
Amino acid structure sequence Ac-EEMQRRYGRKKRRQRRR, the connection of remaining amino acid has been connect successively, to complete the peptide chain
Extension.(9) acetic anhydride of 3 times of resin moles, the pyridine of 3 times of resin moles, reaction is added in reaction column again
30min.After acetyl group (AC) connects, the synthesis of whole peptide is completed with regard to reaction.Into the last contraction phase, since AC does not have
Fmoc is protected, so herein just without deprotection.The contraction of peptide：With DMF washing reactions 3 times, DCM washing reactions 3 times, methanol
Washing reaction 3 times, finally drains peptide resin.(10) amino acid side chain deprotection and the cutting of resin use the side chain protection contained
Base has：Pbf, Trt, Boc, Tbu, these types of protecting group is unstable under acidic condition, and that cutting resin is TFA, so
Deprotection and cutting resin can be carried out at the same time.Configuration cuts liquid 15ml, the volume ratio of wherein each component are：TFA (94.5%),
Water (2%), EDT (2.5%), TIS (1%).Resin is fitted into flask, (30 DEG C) oscillation 2h of constant temperature.By lysate nitrogen
It dries up, is subsequently poured into centrifuge tube as possible, be poured slowly into ether.It seals and is put into centrifuge and centrifuge 5min, outwell supernatant,
Lower section is white solid.6 times are washed with ether, then room temperature volatilizes to get crude product peptide again.(11) use the progress of HPLC methods pure
Change, i.e.,：1. dissolving：Crude product peptide is put into vessel, is completely dissolved with the acetonitrile solution of 30-50ml a concentration of 50%, it can be with
Slightly ultrasound 2min.2. filtering：With 0.45 μm of membrane filtration lysate.3. analyzing：Take 3 μ l solution analysis levels HPLC analyses thick
Product are in order to subsequently preparing.Mobile phase is water and acetonitrile, time 30min, and gradient elution first balances HPLC with start gradient
Then sample introduction, start gradient are 5min：Water 95%, acetonitrile 5%, terminating gradient is：Water 5%, acetonitrile 95%.4. preparing：It will be molten
The sample solved does sample introduction preparation.It prepares HPLC and balances 10min, start gradient is：Water 95%, acetonitrile 5%, terminating gradient is：
Water 25%, acetonitrile 75%, gradient timetable 40min.Collect the sample come out from detector.5. identifying：The sample of collection is carried out
Sample and do purity and mass spectrographic identification.
Second, the raw material prescription and its ratio of a kind of cell-penetrating peptide-acetyl group Argireline nano-emulsion of design simultaneously carry out laboratory system
It is standby.The preparation method of this cell-penetrating peptide-acetyl group Argireline nano-emulsion is as follows：
1) proportioning of cell-penetrating peptide-acetyl group Argireline nanometer dairy milk starting material is pressed, isopropyl myristate is spare as oil phase (A)；
2) cell-penetrating peptide-acetyl group Argireline, appropriate distilled water are mixed and is allowed to be completely dissolved, then distilled water is added to be allowed to as water phase
(C) spare；
3) Labraso, polyglyceryl fatty acid ester are equal to 3: 1 ratio, are placed among another container,
It stirs, is allowed to form Labraso, polyglyceryl fatty acid ester mixture (B) spare；
4) again by oil phase (A): Labraso, polyglyceryl fatty acid ester mixture (B): water phase (C) is equal to
7.66: 23.37: 68.97 ratio, take respectively Labraso, polyglyceryl fatty acid ester mixture (B),
Water phase (C) is added in oil phase (A) container according to this；
5) time constant-temperature magnetic stirring apparatus at ambient temperature, is selected, and with 200rpmmin-1Rotating speed magnetic agitation
30min, you can obtain this cell-penetrating peptide-acetyl group Argireline nano-emulsion.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201810461431.6A CN108714111B (en) | 2018-05-05 | 2018-05-05 | Cell-penetrating peptide-acetyl hexapeptide nanoemulsion and preparation method thereof |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201810461431.6A CN108714111B (en) | 2018-05-05 | 2018-05-05 | Cell-penetrating peptide-acetyl hexapeptide nanoemulsion and preparation method thereof |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN108714111A true CN108714111A (en) | 2018-10-30 |
| CN108714111B CN108714111B (en) | 2022-05-24 |
Family
ID=63900006
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201810461431.6A Expired - Fee Related CN108714111B (en) | 2018-05-05 | 2018-05-05 | Cell-penetrating peptide-acetyl hexapeptide nanoemulsion and preparation method thereof |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN108714111B (en) |
Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN110882178A (en) * | 2019-12-19 | 2020-03-17 | 深圳市维琪医药研发有限公司 | Water-in-oil system whitening polypeptide composition |
| CN114044803A (en) * | 2022-01-12 | 2022-02-15 | 浙江湃肽生物有限公司深圳分公司 | Acetyl hexapeptide-1 derivative and application thereof |
| CN114106205A (en) * | 2021-11-30 | 2022-03-01 | 滨州医学院 | Preparation of mesenchymal stem cell exosome and composition and application of composition in cosmetics |
| CN115144508A (en) * | 2022-09-02 | 2022-10-04 | 广州市乾相生物科技有限公司 | HPLC separation method suitable for multiple water-soluble peptides |
| CN115637279A (en) * | 2022-12-01 | 2023-01-24 | 翔鹏(北京)生物科技有限公司 | Recombinant human collagen for repairing skin and preparation method thereof |
Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101214200A (en) * | 2008-01-21 | 2008-07-09 | 丁克祥 | Skin externally-applied liquid state and solid state kreotoxin (liushengtai) composite nano emulsion and preparation thereof |
| CN101711874A (en) * | 2008-10-08 | 2010-05-26 | 广州暨南大学医药生物技术研究开发中心 | Application of cell penetrating peptide Tat-mediated growth factor in transdermal transfer |
| CN101874762A (en) * | 2009-12-08 | 2010-11-03 | 董萍 | Skin anti-aging composite nano emulsion and preparation method thereof |
| CN102988193A (en) * | 2011-09-19 | 2013-03-27 | 鼎翔生医科技有限公司 | Medical beautifying polypeptide capable of promoting cell regeneration and delaying skin aging |
| CN106916207A (en) * | 2017-01-20 | 2017-07-04 | 肽泽（武汉）生物科技有限公司 | The method of the plasmid DNA transfection of cell-penetrating peptide hPP chol, production and its mediation |
-
2018
- 2018-05-05 CN CN201810461431.6A patent/CN108714111B/en not_active Expired - Fee Related
Patent Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101214200A (en) * | 2008-01-21 | 2008-07-09 | 丁克祥 | Skin externally-applied liquid state and solid state kreotoxin (liushengtai) composite nano emulsion and preparation thereof |
| CN101711874A (en) * | 2008-10-08 | 2010-05-26 | 广州暨南大学医药生物技术研究开发中心 | Application of cell penetrating peptide Tat-mediated growth factor in transdermal transfer |
| CN101874762A (en) * | 2009-12-08 | 2010-11-03 | 董萍 | Skin anti-aging composite nano emulsion and preparation method thereof |
| CN102988193A (en) * | 2011-09-19 | 2013-03-27 | 鼎翔生医科技有限公司 | Medical beautifying polypeptide capable of promoting cell regeneration and delaying skin aging |
| CN106916207A (en) * | 2017-01-20 | 2017-07-04 | 肽泽（武汉）生物科技有限公司 | The method of the plasmid DNA transfection of cell-penetrating peptide hPP chol, production and its mediation |
Non-Patent Citations (2)
| Title |
|---|
| 何俊等: "《促进药物经皮吸收的促渗肽研究进展》", 《中国药学杂志》 * |
| 谢文林等: "《穿膜肽修饰胸腺五肽类似物的合成和生物活性研究》", 《华中师范大学学报（自然科学版）》 * |
Cited By (8)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN110882178A (en) * | 2019-12-19 | 2020-03-17 | 深圳市维琪医药研发有限公司 | Water-in-oil system whitening polypeptide composition |
| CN110882178B (en) * | 2019-12-19 | 2023-11-24 | 深圳市维琪科技股份有限公司 | Water-in-oil system whitening polypeptide composition |
| CN114106205A (en) * | 2021-11-30 | 2022-03-01 | 滨州医学院 | Preparation of mesenchymal stem cell exosome and composition and application of composition in cosmetics |
| CN114106205B (en) * | 2021-11-30 | 2023-10-10 | 滨州医学院 | Preparation of mesenchymal stem cell exosomes and compositions and application in cosmetics |
| CN114044803A (en) * | 2022-01-12 | 2022-02-15 | 浙江湃肽生物有限公司深圳分公司 | Acetyl hexapeptide-1 derivative and application thereof |
| CN115144508A (en) * | 2022-09-02 | 2022-10-04 | 广州市乾相生物科技有限公司 | HPLC separation method suitable for multiple water-soluble peptides |
| CN115144508B (en) * | 2022-09-02 | 2022-12-13 | 广州市乾相生物科技有限公司 | HPLC separation method suitable for multiple water-soluble peptides |
| CN115637279A (en) * | 2022-12-01 | 2023-01-24 | 翔鹏(北京)生物科技有限公司 | Recombinant human collagen for repairing skin and preparation method thereof |
Also Published As
| Publication number | Publication date |
|---|---|
| CN108714111B (en) | 2022-05-24 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN108714111A (en) | A kind of cell-penetrating peptide-acetyl group Argireline nano-emulsion and preparation method thereof | |
| JP7227903B2 (en) | Compounds useful for the treatment and/or care of skin, hair, nails and/or mucous membranes | |
| CN103314005B (en) | Useful peptide and their purposes in cosmetic composition or Pharmaceutical composition in skin and/or the treatment of mucosa and/or nursing | |
| Lintner et al. | Biologically active peptides: from a laboratory bench curiosity to a functional skin care product | |
| CN110237022A (en) | A kind of freeze-dried powder, solvent and its application | |
| RU2557401C2 (en) | Peptides, applied in skin, mucosa and/or hair treatment and/or care, and their application in cosmetic or pharmaceutical compositions | |
| EP2456457B1 (en) | Compounds which inhibit muscle contraction | |
| CN101541831A (en) | Neuronal exocytosis inhibiting peptides | |
| KR20130135819A (en) | Skin antiaging treatment | |
| CN105408345A (en) | Compounds useful in treatment and/or care of skin and cosmetic or pharmaceutical compositions thereof | |
| JP2020500155A (en) | Compounds useful in the treatment and / or care of skin, hair, nails and / or mucous membranes | |
| CN101214200A (en) | Skin externally-applied liquid state and solid state kreotoxin (liushengtai) composite nano emulsion and preparation thereof | |
| CN105999535A (en) | Application of sponge spicule as skin physical penetration enhancer | |
| JP5247676B2 (en) | Synthetic peptides useful in skin treatment and their use in cosmetic or dermopharmaceutical compositions | |
| Ledwoń et al. | Peptides as active ingredients: a challenge for cosmeceutical industry | |
| CN102292126B (en) | For treating Hyperpigmented compositions and method | |
| CN106963688A (en) | It is a kind of containing plant extracts complex polypeptide liposome composition and preparation method thereof and in the application of anti-wrinkle product | |
| US20050124545A1 (en) | Cosmetic or parmaceutical composition containing peptides | |
| BR112020000123A2 (en) | compound, use of a compound, cosmetic composition, and, method for non-therapeutic cosmetic care and / or treatment of skin, hair, nails and / or mucous membranes | |
| EP2164858A1 (en) | Novel compounds, use thereof in cosmetic and cosmeceutic applications, and compositions comprising same | |
| CN104321049B (en) | The compound of inhibitory neuron exocytosis | |
| CN110526957A (en) | Premeabilisation of cells enhances peptide, composition and its application | |
| TW202333650A (en) | Methods and compositions for cosmetic applications | |
| CN111961119B (en) | Application of polypeptide in preparation of medicine or cosmetic for promoting collagen secretion | |
| WO2020254690A1 (en) | Composition modulating botulinum neurotoxin effect |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| GR01 | Patent grant | ||
| GR01 | Patent grant | ||
| CF01 | Termination of patent right due to non-payment of annual fee |
Granted publication date: 20220524 |