CN117304368B - Composition of sodium hyaluronate and collagen pentapeptide and application thereof - Google Patents
Composition of sodium hyaluronate and collagen pentapeptide and application thereof Download PDFInfo
- Publication number
- CN117304368B CN117304368B CN202311606423.3A CN202311606423A CN117304368B CN 117304368 B CN117304368 B CN 117304368B CN 202311606423 A CN202311606423 A CN 202311606423A CN 117304368 B CN117304368 B CN 117304368B
- Authority
- CN
- China
- Prior art keywords
- sodium hyaluronate
- derivative
- collagen
- composition
- thousand
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Active
Links
- YWIVKILSMZOHHF-QJZPQSOGSA-N sodium;(2s,3s,4s,5r,6r)-6-[(2s,3r,4r,5s,6r)-3-acetamido-2-[(2s,3s,4r,5r,6r)-6-[(2r,3r,4r,5s,6r)-3-acetamido-2,5-dihydroxy-6-(hydroxymethyl)oxan-4-yl]oxy-2-carboxy-4,5-dihydroxyoxan-3-yl]oxy-5-hydroxy-6-(hydroxymethyl)oxan-4-yl]oxy-3,4,5-trihydroxyoxane-2- Chemical group [Na+].CC(=O)N[C@H]1[C@H](O)O[C@H](CO)[C@@H](O)[C@@H]1O[C@H]1[C@H](O)[C@@H](O)[C@H](O[C@H]2[C@@H]([C@@H](O[C@H]3[C@@H]([C@@H](O)[C@H](O)[C@H](O3)C(O)=O)O)[C@H](O)[C@@H](CO)O2)NC(C)=O)[C@@H](C(O)=O)O1 YWIVKILSMZOHHF-QJZPQSOGSA-N 0.000 title claims abstract description 118
- 229920002385 Sodium hyaluronate Polymers 0.000 title claims abstract description 77
- 229940010747 sodium hyaluronate Drugs 0.000 title claims abstract description 77
- 102000008186 Collagen Human genes 0.000 title claims abstract description 51
- 108010035532 Collagen Proteins 0.000 title claims abstract description 51
- 229920001436 collagen Polymers 0.000 title claims abstract description 51
- 239000000203 mixture Substances 0.000 title claims abstract description 43
- FBPFZTCFMRRESA-KVTDHHQDSA-N D-Mannitol Chemical compound OC[C@@H](O)[C@@H](O)[C@H](O)[C@H](O)CO FBPFZTCFMRRESA-KVTDHHQDSA-N 0.000 claims abstract description 46
- 239000002537 cosmetic Substances 0.000 claims abstract description 10
- KIUKXJAPPMFGSW-DNGZLQJQSA-N (2S,3S,4S,5R,6R)-6-[(2S,3R,4R,5S,6R)-3-Acetamido-2-[(2S,3S,4R,5R,6R)-6-[(2R,3R,4R,5S,6R)-3-acetamido-2,5-dihydroxy-6-(hydroxymethyl)oxan-4-yl]oxy-2-carboxy-4,5-dihydroxyoxan-3-yl]oxy-5-hydroxy-6-(hydroxymethyl)oxan-4-yl]oxy-3,4,5-trihydroxyoxane-2-carboxylic acid Chemical class CC(=O)N[C@H]1[C@H](O)O[C@H](CO)[C@@H](O)[C@@H]1O[C@H]1[C@H](O)[C@@H](O)[C@H](O[C@H]2[C@@H]([C@@H](O[C@H]3[C@@H]([C@@H](O)[C@H](O)[C@H](O3)C(O)=O)O)[C@H](O)[C@@H](CO)O2)NC(C)=O)[C@@H](C(O)=O)O1 KIUKXJAPPMFGSW-DNGZLQJQSA-N 0.000 claims description 35
- 238000002360 preparation method Methods 0.000 claims description 20
- NHGXDBSUJJNIRV-UHFFFAOYSA-M tetrabutylammonium chloride Chemical compound [Cl-].CCCC[N+](CCCC)(CCCC)CCCC NHGXDBSUJJNIRV-UHFFFAOYSA-M 0.000 claims description 18
- 238000004132 cross linking Methods 0.000 claims description 16
- PPXLQETZAQHSSL-UHFFFAOYSA-N 2-[4-(3-chloropropyl)piperazin-1-yl]ethanol Chemical compound OCCN1CCN(CCCCl)CC1 PPXLQETZAQHSSL-UHFFFAOYSA-N 0.000 claims description 15
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 14
- AGYUOJIYYGGHKV-UHFFFAOYSA-N 1,2-bis(2-chloroethoxy)ethane Chemical compound ClCCOCCOCCCl AGYUOJIYYGGHKV-UHFFFAOYSA-N 0.000 claims description 12
- -1 5-chlorosulfonyl-2-hydroxybenzoic acid modified mannitol Chemical class 0.000 claims description 6
- 239000000843 powder Substances 0.000 claims description 5
- 239000007864 aqueous solution Substances 0.000 claims description 4
- 239000003814 drug Substances 0.000 claims description 3
- 239000000945 filler Substances 0.000 claims description 3
- 239000000126 substance Substances 0.000 claims description 3
- FBPFZTCFMRRESA-FSIIMWSLSA-N D-Glucitol Natural products OC[C@H](O)[C@H](O)[C@@H](O)[C@H](O)CO FBPFZTCFMRRESA-FSIIMWSLSA-N 0.000 claims description 2
- 229920002307 Dextran Polymers 0.000 claims description 2
- 239000004615 ingredient Substances 0.000 claims description 2
- 238000002347 injection Methods 0.000 claims description 2
- 239000007924 injection Substances 0.000 claims description 2
- 239000000600 sorbitol Substances 0.000 claims description 2
- 235000010355 mannitol Nutrition 0.000 abstract description 21
- 229930195725 Mannitol Natural products 0.000 abstract description 17
- 239000000594 mannitol Substances 0.000 abstract description 17
- 230000008929 regeneration Effects 0.000 abstract description 14
- 238000011069 regeneration method Methods 0.000 abstract description 14
- 230000002195 synergetic effect Effects 0.000 abstract description 13
- 230000004936 stimulating effect Effects 0.000 abstract description 7
- 102000004196 processed proteins & peptides Human genes 0.000 abstract description 6
- 108090000765 processed proteins & peptides Proteins 0.000 abstract description 6
- 239000011243 crosslinked material Substances 0.000 abstract description 5
- 229920001184 polypeptide Polymers 0.000 abstract description 5
- 238000012360 testing method Methods 0.000 description 43
- 230000000694 effects Effects 0.000 description 34
- 229920002674 hyaluronan Polymers 0.000 description 32
- 229960003160 hyaluronic acid Drugs 0.000 description 32
- 230000014509 gene expression Effects 0.000 description 25
- 239000000523 sample Substances 0.000 description 20
- 238000003756 stirring Methods 0.000 description 14
- 102000004887 Transforming Growth Factor beta Human genes 0.000 description 13
- 108090001012 Transforming Growth Factor beta Proteins 0.000 description 13
- ZRKFYGHZFMAOKI-QMGMOQQFSA-N tgfbeta Chemical compound C([C@H](NC(=O)[C@H](C(C)C)NC(=O)CNC(=O)[C@H](CCC(O)=O)NC(=O)[C@H](CCCNC(N)=N)NC(=O)[C@H](CC(N)=O)NC(=O)[C@H](CC(C)C)NC(=O)[C@H]([C@@H](C)O)NC(=O)[C@H](CCC(O)=O)NC(=O)[C@H]([C@@H](C)O)NC(=O)[C@H](CC(C)C)NC(=O)CNC(=O)[C@H](C)NC(=O)[C@H](CO)NC(=O)[C@H](CCC(N)=O)NC(=O)[C@@H](NC(=O)[C@H](C)NC(=O)[C@H](C)NC(=O)[C@@H](NC(=O)[C@H](CC(C)C)NC(=O)[C@@H](N)CCSC)C(C)C)[C@@H](C)CC)C(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@@H](C(C)C)C(=O)N[C@@H](CC=1C=CC=CC=1)C(=O)N[C@@H](C)C(=O)N1[C@@H](CCC1)C(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CC(N)=O)C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H](C)C(=O)N[C@@H](CC=1C=CC=CC=1)C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](C)C(=O)N[C@@H](CC(C)C)C(=O)N1[C@@H](CCC1)C(=O)N1[C@@H](CCC1)C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CO)C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CC(C)C)C(O)=O)C1=CC=C(O)C=C1 ZRKFYGHZFMAOKI-QMGMOQQFSA-N 0.000 description 13
- 238000004458 analytical method Methods 0.000 description 12
- 102000002274 Matrix Metalloproteinases Human genes 0.000 description 11
- 108010000684 Matrix Metalloproteinases Proteins 0.000 description 11
- JUJWROOIHBZHMG-UHFFFAOYSA-N Pyridine Chemical compound C1=CC=NC=C1 JUJWROOIHBZHMG-UHFFFAOYSA-N 0.000 description 10
- 210000003491 skin Anatomy 0.000 description 10
- 230000005764 inhibitory process Effects 0.000 description 9
- SJXHSFSHNKFRLN-UHFFFAOYSA-N 5-chlorosulfonyl-2-hydroxybenzoic acid Chemical compound OC(=O)C1=CC(S(Cl)(=O)=O)=CC=C1O SJXHSFSHNKFRLN-UHFFFAOYSA-N 0.000 description 8
- VEXZGXHMUGYJMC-UHFFFAOYSA-N Hydrochloric acid Chemical compound Cl VEXZGXHMUGYJMC-UHFFFAOYSA-N 0.000 description 8
- 238000006243 chemical reaction Methods 0.000 description 8
- 239000005457 ice water Substances 0.000 description 8
- 108090000623 proteins and genes Proteins 0.000 description 8
- 108010022452 Collagen Type I Proteins 0.000 description 7
- 102000010834 Extracellular Matrix Proteins Human genes 0.000 description 7
- 108010037362 Extracellular Matrix Proteins Proteins 0.000 description 7
- 238000010521 absorption reaction Methods 0.000 description 7
- 210000002744 extracellular matrix Anatomy 0.000 description 7
- 239000000243 solution Substances 0.000 description 7
- 230000000638 stimulation Effects 0.000 description 7
- 239000004971 Cross linker Substances 0.000 description 6
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 description 6
- ZMXDDKWLCZADIW-UHFFFAOYSA-N N,N-Dimethylformamide Chemical compound CN(C)C=O ZMXDDKWLCZADIW-UHFFFAOYSA-N 0.000 description 6
- 238000010998 test method Methods 0.000 description 6
- 102000012422 Collagen Type I Human genes 0.000 description 5
- 239000000306 component Substances 0.000 description 5
- 231100000263 cytotoxicity test Toxicity 0.000 description 5
- 210000002950 fibroblast Anatomy 0.000 description 5
- 239000007788 liquid Substances 0.000 description 5
- UMJSCPRVCHMLSP-UHFFFAOYSA-N pyridine Natural products COC1=CC=CN=C1 UMJSCPRVCHMLSP-UHFFFAOYSA-N 0.000 description 5
- ZAMOUSCENKQFHK-UHFFFAOYSA-N Chlorine atom Chemical compound [Cl] ZAMOUSCENKQFHK-UHFFFAOYSA-N 0.000 description 4
- 108010069502 Collagen Type III Proteins 0.000 description 4
- 238000002835 absorbance Methods 0.000 description 4
- 230000003712 anti-aging effect Effects 0.000 description 4
- 239000000460 chlorine Substances 0.000 description 4
- 229910052801 chlorine Inorganic materials 0.000 description 4
- 238000002784 cytotoxicity assay Methods 0.000 description 4
- 238000001514 detection method Methods 0.000 description 4
- 238000002156 mixing Methods 0.000 description 4
- 230000004048 modification Effects 0.000 description 4
- 238000012986 modification Methods 0.000 description 4
- 230000001105 regulatory effect Effects 0.000 description 4
- 238000001291 vacuum drying Methods 0.000 description 4
- 102000001187 Collagen Type III Human genes 0.000 description 3
- IAZDPXIOMUYVGZ-UHFFFAOYSA-N Dimethylsulphoxide Chemical compound CS(C)=O IAZDPXIOMUYVGZ-UHFFFAOYSA-N 0.000 description 3
- 101001013150 Homo sapiens Interstitial collagenase Proteins 0.000 description 3
- 102000000380 Matrix Metalloproteinase 1 Human genes 0.000 description 3
- YXFVVABEGXRONW-UHFFFAOYSA-N Toluene Chemical compound CC1=CC=CC=C1 YXFVVABEGXRONW-UHFFFAOYSA-N 0.000 description 3
- 230000015572 biosynthetic process Effects 0.000 description 3
- KRKNYBCHXYNGOX-UHFFFAOYSA-N citric acid Chemical compound OC(=O)CC(O)(C(O)=O)CC(O)=O KRKNYBCHXYNGOX-UHFFFAOYSA-N 0.000 description 3
- 150000001875 compounds Chemical class 0.000 description 3
- 231100000433 cytotoxic Toxicity 0.000 description 3
- 230000001472 cytotoxic effect Effects 0.000 description 3
- 230000002500 effect on skin Effects 0.000 description 3
- 238000005516 engineering process Methods 0.000 description 3
- 238000002329 infrared spectrum Methods 0.000 description 3
- 238000000034 method Methods 0.000 description 3
- 150000003254 radicals Chemical class 0.000 description 3
- 239000012488 sample solution Substances 0.000 description 3
- 238000001179 sorption measurement Methods 0.000 description 3
- 238000003786 synthesis reaction Methods 0.000 description 3
- 101100328883 Arabidopsis thaliana COL1 gene Proteins 0.000 description 2
- 101100328890 Arabidopsis thaliana COL3 gene Proteins 0.000 description 2
- 102000000503 Collagen Type II Human genes 0.000 description 2
- 108010041390 Collagen Type II Proteins 0.000 description 2
- 230000009471 action Effects 0.000 description 2
- 230000032683 aging Effects 0.000 description 2
- 230000003110 anti-inflammatory effect Effects 0.000 description 2
- 238000003556 assay Methods 0.000 description 2
- 238000013329 compounding Methods 0.000 description 2
- 238000001816 cooling Methods 0.000 description 2
- 238000007405 data analysis Methods 0.000 description 2
- 210000004207 dermis Anatomy 0.000 description 2
- 150000002016 disaccharides Chemical class 0.000 description 2
- 230000001815 facial effect Effects 0.000 description 2
- 230000002349 favourable effect Effects 0.000 description 2
- 238000001914 filtration Methods 0.000 description 2
- 230000007760 free radical scavenging Effects 0.000 description 2
- 239000001963 growth medium Substances 0.000 description 2
- 238000010438 heat treatment Methods 0.000 description 2
- 238000004519 manufacturing process Methods 0.000 description 2
- 239000000463 material Substances 0.000 description 2
- 125000000325 methylidene group Chemical group [H]C([H])=* 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
- 230000035515 penetration Effects 0.000 description 2
- 230000003716 rejuvenation Effects 0.000 description 2
- 230000008439 repair process Effects 0.000 description 2
- 238000011160 research Methods 0.000 description 2
- 210000000434 stratum corneum Anatomy 0.000 description 2
- 239000000725 suspension Substances 0.000 description 2
- 230000009897 systematic effect Effects 0.000 description 2
- 230000000699 topical effect Effects 0.000 description 2
- 238000005406 washing Methods 0.000 description 2
- 108091032973 (ribonucleotides)n+m Proteins 0.000 description 1
- 241000283690 Bos taurus Species 0.000 description 1
- 102000029816 Collagenase Human genes 0.000 description 1
- 108060005980 Collagenase Proteins 0.000 description 1
- FBPFZTCFMRRESA-JGWLITMVSA-N D-glucitol Chemical compound OC[C@H](O)[C@@H](O)[C@H](O)[C@H](O)CO FBPFZTCFMRRESA-JGWLITMVSA-N 0.000 description 1
- 208000005422 Foreign-Body reaction Diseases 0.000 description 1
- IAJILQKETJEXLJ-UHFFFAOYSA-N Galacturonsaeure Natural products O=CC(O)C(O)C(O)C(O)C(O)=O IAJILQKETJEXLJ-UHFFFAOYSA-N 0.000 description 1
- 241000282412 Homo Species 0.000 description 1
- 108010016113 Matrix Metalloproteinase 1 Proteins 0.000 description 1
- 241001465754 Metazoa Species 0.000 description 1
- OVRNDRQMDRJTHS-UHFFFAOYSA-N N-acelyl-D-glucosamine Natural products CC(=O)NC1C(O)OC(CO)C(O)C1O OVRNDRQMDRJTHS-UHFFFAOYSA-N 0.000 description 1
- OVRNDRQMDRJTHS-FMDGEEDCSA-N N-acetyl-beta-D-glucosamine Chemical compound CC(=O)N[C@H]1[C@H](O)O[C@H](CO)[C@@H](O)[C@@H]1O OVRNDRQMDRJTHS-FMDGEEDCSA-N 0.000 description 1
- 206010028980 Neoplasm Diseases 0.000 description 1
- 102000035195 Peptidases Human genes 0.000 description 1
- 108091005804 Peptidases Proteins 0.000 description 1
- 108010076504 Protein Sorting Signals Proteins 0.000 description 1
- AEMOLEFTQBMNLQ-WAXACMCWSA-N alpha-D-glucuronic acid Chemical compound O[C@H]1O[C@H](C(O)=O)[C@@H](O)[C@H](O)[C@H]1O AEMOLEFTQBMNLQ-WAXACMCWSA-N 0.000 description 1
- 150000001413 amino acids Chemical group 0.000 description 1
- 230000003321 amplification Effects 0.000 description 1
- 230000003064 anti-oxidating effect Effects 0.000 description 1
- 230000003796 beauty Effects 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 230000031018 biological processes and functions Effects 0.000 description 1
- 230000006696 biosynthetic metabolic pathway Effects 0.000 description 1
- 201000011510 cancer Diseases 0.000 description 1
- 125000003178 carboxy group Chemical group [H]OC(*)=O 0.000 description 1
- 230000015556 catabolic process Effects 0.000 description 1
- 230000033077 cellular process Effects 0.000 description 1
- 230000008859 change Effects 0.000 description 1
- 238000012512 characterization method Methods 0.000 description 1
- 238000007385 chemical modification Methods 0.000 description 1
- 230000011382 collagen catabolic process Effects 0.000 description 1
- 229960002424 collagenase Drugs 0.000 description 1
- 239000008358 core component Substances 0.000 description 1
- 231100000135 cytotoxicity Toxicity 0.000 description 1
- 230000003013 cytotoxicity Effects 0.000 description 1
- 230000007547 defect Effects 0.000 description 1
- 238000006731 degradation reaction Methods 0.000 description 1
- 238000003745 diagnosis Methods 0.000 description 1
- 238000009792 diffusion process Methods 0.000 description 1
- 239000006185 dispersion Substances 0.000 description 1
- 231100000673 dose–response relationship Toxicity 0.000 description 1
- HHEAADYXPMHMCT-UHFFFAOYSA-N dpph Chemical compound [O-][N+](=O)C1=CC([N+](=O)[O-])=CC([N+]([O-])=O)=C1[N]N(C=1C=CC=CC=1)C1=CC=CC=C1 HHEAADYXPMHMCT-UHFFFAOYSA-N 0.000 description 1
- 238000010894 electron beam technology Methods 0.000 description 1
- 230000002708 enhancing effect Effects 0.000 description 1
- 238000002474 experimental method Methods 0.000 description 1
- 238000011049 filling Methods 0.000 description 1
- 230000012010 growth Effects 0.000 description 1
- 239000003102 growth factor Substances 0.000 description 1
- 230000013632 homeostatic process Effects 0.000 description 1
- 230000036571 hydration Effects 0.000 description 1
- 238000006703 hydration reaction Methods 0.000 description 1
- 230000002519 immonomodulatory effect Effects 0.000 description 1
- 238000002513 implantation Methods 0.000 description 1
- 230000002401 inhibitory effect Effects 0.000 description 1
- 238000011835 investigation Methods 0.000 description 1
- FZWBNHMXJMCXLU-BLAUPYHCSA-N isomaltotriose Chemical compound O[C@@H]1[C@@H](O)[C@H](O)[C@@H](CO)O[C@@H]1OC[C@@H]1[C@@H](O)[C@H](O)[C@@H](O)[C@@H](OC[C@@H](O)[C@@H](O)[C@H](O)[C@@H](O)C=O)O1 FZWBNHMXJMCXLU-BLAUPYHCSA-N 0.000 description 1
- 238000009630 liquid culture Methods 0.000 description 1
- 239000002075 main ingredient Substances 0.000 description 1
- 210000001161 mammalian embryo Anatomy 0.000 description 1
- 230000007246 mechanism Effects 0.000 description 1
- 239000002609 medium Substances 0.000 description 1
- 108020004999 messenger RNA Proteins 0.000 description 1
- 230000004060 metabolic process Effects 0.000 description 1
- 125000002496 methyl group Chemical group [H]C([H])([H])* 0.000 description 1
- 239000004005 microsphere Substances 0.000 description 1
- 229950006780 n-acetylglucosamine Drugs 0.000 description 1
- 230000007935 neutral effect Effects 0.000 description 1
- 231100001083 no cytotoxicity Toxicity 0.000 description 1
- 238000003199 nucleic acid amplification method Methods 0.000 description 1
- 108020004707 nucleic acids Proteins 0.000 description 1
- 102000039446 nucleic acids Human genes 0.000 description 1
- 150000007523 nucleic acids Chemical class 0.000 description 1
- 230000003647 oxidation Effects 0.000 description 1
- 238000007254 oxidation reaction Methods 0.000 description 1
- 125000001997 phenyl group Chemical group [H]C1=C([H])C([H])=C(*)C([H])=C1[H] 0.000 description 1
- 230000001376 precipitating effect Effects 0.000 description 1
- 238000001556 precipitation Methods 0.000 description 1
- 230000008569 process Effects 0.000 description 1
- 230000001737 promoting effect Effects 0.000 description 1
- 230000017854 proteolysis Effects 0.000 description 1
- 230000006337 proteolytic cleavage Effects 0.000 description 1
- 229940024999 proteolytic enzymes for treatment of wounds and ulcers Drugs 0.000 description 1
- 238000007634 remodeling Methods 0.000 description 1
- 210000002966 serum Anatomy 0.000 description 1
- 230000019491 signal transduction Effects 0.000 description 1
- 239000002904 solvent Substances 0.000 description 1
- 210000002784 stomach Anatomy 0.000 description 1
- 238000007920 subcutaneous administration Methods 0.000 description 1
- 238000006467 substitution reaction Methods 0.000 description 1
- 238000000967 suction filtration Methods 0.000 description 1
- 238000012385 systemic delivery Methods 0.000 description 1
- 238000010257 thawing Methods 0.000 description 1
- 230000017423 tissue regeneration Effects 0.000 description 1
- 230000001131 transforming effect Effects 0.000 description 1
- 238000002834 transmittance Methods 0.000 description 1
- 230000003827 upregulation Effects 0.000 description 1
- 238000003828 vacuum filtration Methods 0.000 description 1
- 238000012795 verification Methods 0.000 description 1
- 230000029663 wound healing Effects 0.000 description 1
Classifications
-
- C—CHEMISTRY; METALLURGY
- C08—ORGANIC MACROMOLECULAR COMPOUNDS; THEIR PREPARATION OR CHEMICAL WORKING-UP; COMPOSITIONS BASED THEREON
- C08B—POLYSACCHARIDES; DERIVATIVES THEREOF
- C08B37/00—Preparation of polysaccharides not provided for in groups C08B1/00 - C08B35/00; Derivatives thereof
- C08B37/006—Heteroglycans, i.e. polysaccharides having more than one sugar residue in the main chain in either alternating or less regular sequence; Gellans; Succinoglycans; Arabinogalactans; Tragacanth or gum tragacanth or traganth from Astragalus; Gum Karaya from Sterculia urens; Gum Ghatti from Anogeissus latifolia; Derivatives thereof
- C08B37/0063—Glycosaminoglycans or mucopolysaccharides, e.g. keratan sulfate; Derivatives thereof, e.g. fucoidan
- C08B37/0072—Hyaluronic acid, i.e. HA or hyaluronan; Derivatives thereof, e.g. crosslinked hyaluronic acid (hylan) or hyaluronates
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K31/00—Medicinal preparations containing organic active ingredients
- A61K31/70—Carbohydrates; Sugars; Derivatives thereof
- A61K31/715—Polysaccharides, i.e. having more than five saccharide radicals attached to each other by glycosidic linkages; Derivatives thereof, e.g. ethers, esters
- A61K31/726—Glycosaminoglycans, i.e. mucopolysaccharides
- A61K31/728—Hyaluronic acid
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K38/00—Medicinal preparations containing peptides
- A61K38/04—Peptides having up to 20 amino acids in a fully defined sequence; Derivatives thereof
- A61K38/08—Peptides having 5 to 11 amino acids
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K8/00—Cosmetics or similar toiletry preparations
- A61K8/18—Cosmetics or similar toiletry preparations characterised by the composition
- A61K8/30—Cosmetics or similar toiletry preparations characterised by the composition containing organic compounds
- A61K8/33—Cosmetics or similar toiletry preparations characterised by the composition containing organic compounds containing oxygen
- A61K8/34—Alcohols
- A61K8/345—Alcohols containing more than one hydroxy group
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K8/00—Cosmetics or similar toiletry preparations
- A61K8/18—Cosmetics or similar toiletry preparations characterised by the composition
- A61K8/30—Cosmetics or similar toiletry preparations characterised by the composition containing organic compounds
- A61K8/64—Proteins; Peptides; Derivatives or degradation products thereof
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K8/00—Cosmetics or similar toiletry preparations
- A61K8/18—Cosmetics or similar toiletry preparations characterised by the composition
- A61K8/72—Cosmetics or similar toiletry preparations characterised by the composition containing organic macromolecular compounds
- A61K8/73—Polysaccharides
-
- 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
- A61K8/72—Cosmetics or similar toiletry preparations characterised by the composition containing organic macromolecular compounds
- A61K8/73—Polysaccharides
- A61K8/735—Mucopolysaccharides, e.g. hyaluronic acid; Derivatives thereof
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
- A61L27/00—Materials for grafts or prostheses or for coating grafts or prostheses
- A61L27/14—Macromolecular materials
- A61L27/26—Mixtures of macromolecular compounds
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P17/00—Drugs for dermatological disorders
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61Q—SPECIFIC USE OF COSMETICS OR SIMILAR TOILETRY PREPARATIONS
- A61Q19/00—Preparations for care of the skin
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61Q—SPECIFIC USE OF COSMETICS OR SIMILAR TOILETRY PREPARATIONS
- A61Q19/00—Preparations for care of the skin
- A61Q19/08—Anti-ageing preparations
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
- A61L2400/00—Materials characterised by their function or physical properties
- A61L2400/06—Flowable or injectable implant compositions
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
- A61L2430/00—Materials or treatment for tissue regeneration
- A61L2430/34—Materials or treatment for tissue regeneration for soft tissue reconstruction
-
- 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
- Y02A—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE
- Y02A50/00—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE in human health protection, e.g. against extreme weather
- Y02A50/30—Against vector-borne diseases, e.g. mosquito-borne, fly-borne, tick-borne or waterborne diseases whose impact is exacerbated by climate change
Landscapes
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- General Health & Medical Sciences (AREA)
- Veterinary Medicine (AREA)
- Public Health (AREA)
- Animal Behavior & Ethology (AREA)
- Chemical & Material Sciences (AREA)
- Epidemiology (AREA)
- Dermatology (AREA)
- Medicinal Chemistry (AREA)
- Birds (AREA)
- Pharmacology & Pharmacy (AREA)
- Engineering & Computer Science (AREA)
- Organic Chemistry (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Molecular Biology (AREA)
- Materials Engineering (AREA)
- Emergency Medicine (AREA)
- Gerontology & Geriatric Medicine (AREA)
- Oral & Maxillofacial Surgery (AREA)
- Transplantation (AREA)
- Polymers & Plastics (AREA)
- Gastroenterology & Hepatology (AREA)
- Biochemistry (AREA)
- Immunology (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- General Chemical & Material Sciences (AREA)
- Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
- Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
Abstract
The invention discloses a composition of sodium hyaluronate and collagen pentapeptide and application thereof, and relates to the technical field of polypeptide medical cosmetology cosmetics. The composition comprises sodium hyaluronate or sodium hyaluronate derivatives; the sodium hyaluronate derivative comprises a sodium hyaluronate cross-linked derivative; the crosslinked derivative comprises a mixture crosslinked material of one or more of sodium hyaluronate with single-chain molecular weight of 1-8 ten thousand, 8-15 ten thousand, 15-30 ten thousand, 30-80 ten thousand and 80-200 ten thousand. A composition of sodium hyaluronate and collagen pentapeptide and its application, which combines sodium hyaluronate and collagen pentapeptide, has good ability of stimulating collagen regeneration and produces synergistic effect; and meanwhile, auxiliary mannitol or derivatives thereof are added, so that the collagen regeneration stimulating capability of the composition is further improved.
Description
Technical Field
The invention belongs to the technical field of polypeptide medical beauty cosmetics, and particularly relates to a composition of sodium hyaluronate and collagen pentapeptide and application thereof.
Background
Hyaluronic Acid (HA) consists of a repeating polymeric disaccharide of D-glucuronic acid and N-acetyl-D-glucosamine. Its structural stability depends on the stereochemistry of the disaccharide. The natural abundance, degradability and biocompatibility of HA in animals and humans makes it versatile in regulating various biological processes and maintaining homeostasis, such as skin repair, cancer diagnosis, wound healing, tissue regeneration, anti-inflammatory and immunomodulation. Hyaluronic acid has the ability to capture about 1000 times its weight of water, and its excellent ability to replenish water makes it one of the main ingredients of medical and cosmetic products. In addition, higher concentrations of hyaluronic acid molecules can become entangled, forming a continuous porous network. Such a network structure generates expansion pressure due to the increase of mutual repulsion between molecules and inside the molecules, in an effort to make the self structure relatively loose. When external pressure is applied to the hyaluronic acid network, the molecular network contracts, and when external pressure is withdrawn, the hyaluronic acid network will spring back to its original shape, or acquire a new shape, due to the internal expansion pressure. Because of its remarkable hydrodynamic properties, it can play an important role in the hydration and biomechanical integrity of the dermis when used for facial injection, providing elasticity and extensibility to facial skin.
Collagen pentapeptides are also known as pentapeptides-3 or pentapeptides-4, and the amino acid sequence is lysine-threonine-lysine-serine (KTTKS). Because of its ability to enhance skin remodeling by triggering cellular processes, it can inhibit collagenase activity and increase extracellular matrix (ECM) content, etc., are considered "signal peptides". KTTKS is a subfragment of type I collagen, and was first discovered by a group of researchers at the university of tennessee in 1993 to be the smallest sequence that can retain 80% of the original activity of the large number of parent peptides required to stimulate ECM synthesis. Stimulation with KTTKS is specific for type I collagen expression.
Polypeptides including collagen pentapeptides are generally unsuitable for systemic delivery because they are susceptible to extensive proteolytic degradation, and to extreme pH conditions (stomach). Topical application provides a more direct route to the site of action, and the metabolic processes of the route are greatly reduced, resulting in higher bioavailability. However, their passive penetration and penetration potential on the skin need to be considered when formulating collagen pentapeptides into topical compositions. First, KTTKS is extremely difficult to be absorbed and distributed by the stratum corneum due to its hydrophilic nature. Second, KTTKS is highly likely to interact significantly with proteolytic enzymes during slow diffusion, even if a small fraction diffuses through the stratum corneum. The two above-mentioned points add difficulty to their widespread use. Collagen pentapeptides are usually present in cosmetics in the form of lipophilic modification, and the skin transmittance is increased by modification of palmitoyl groups, but the effective utilization rate of the dermis layer can be only 3%, and the introduction of palmitoyl groups makes the collagen pentapeptides unavailable in conventional water-soluble skin products, and limits the expansion of the application thereof.
Disclosure of Invention
The invention aims to provide a composition of sodium hyaluronate and collagen pentapeptide and application thereof, wherein the composition combines the sodium hyaluronate and the collagen pentapeptide for application, has good collagen regeneration stimulating capability and generates synergistic enhancement effect; mannitol or its derivative is added simultaneously to further promote the collagen regeneration stimulating capacity of the composition.
The technical scheme adopted by the invention for achieving the purpose is as follows:
a sodium hyaluronate derivative comprising a cross-linked sodium hyaluronate derivative; the crosslinked derivative comprises a mixture crosslinked material of one or more of sodium hyaluronate with single-chain molecular weight of 1-8 ten thousand, 8-15 ten thousand, 15-30 ten thousand, 30-80 ten thousand and 80-200 ten thousand.
Specifically, the crosslinking mode of the sodium hyaluronate crosslinking derivative can be selected from the physical crosslinking modes such as traditional thermal crosslinking, freeze thawing crosslinking, electron beam crosslinking, irradiation crosslinking and the like.
A composition comprising the non-crosslinked sodium hyaluronate or crosslinked sodium hyaluronate described above. The invention firstly uses the sodium hyaluronate and the collagen pentapeptide in a combined way to obtain a composition product, and exploits one of core components in the product: the novel application scene of the collagen pentapeptide provides a way that the bioavailability is far higher than that of the traditional external application, HA and KTTKS are combined and applied for systematic research, and the combination of the HA and the KTTKS proves that the HA and the KTTKS have good collagen regeneration stimulating capability and better effect when being combined for use, and the synergic enhancement effect is generated; the composition product can be applied to the fields of subcutaneous implantation, water light, microneedle introduction and the like; and further defines the ultimate biosafety concentration of the collagen pentapeptide, thereby providing guarantee for the safety and effectiveness of the product.
Specifically, the composition further comprises collagen pentapeptide (KTTKS).
Specifically, the composition also comprises auxiliary ingredients including one or more of mannitol, sorbitol and dextran; the use concentration of the auxiliary components is 0.5-10wt%. The invention provides a technical scheme for effectively proportioning sodium hyaluronate, polypeptide and auxiliary materials, which is favorable for skin rejuvenation and has excellent effect. On the premise of taking HA+KTTKS as a main body, mannitol and the like are added as auxiliary components, and the important point is that the auxiliary materials have anti-inflammatory and free radical adsorption capacities, so that an important auxiliary effect is achieved, the action effect of the composition is obviously improved, the expression of TGF-beta is better promoted, the generation of collagen is further promoted, the better MMP-1 inhibition effect is achieved, and the collagen regeneration stimulation capacity is obviously enhanced.
More preferably, mannitol derivatives may be used in the above composition instead of mannitol.
The mannitol derivatives include products of 5-chlorosulfonyl-2-hydroxybenzoic acid modified mannitol. The mannitol derivative prepared by adopting the 5-chlorosulfonyl-2-hydroxybenzoic acid modified mannitol shows better bioactivity, has stronger oxidation resistance and remarkably improves the adsorption removal capability of free radicals; the auxiliary components are added into the composition for compounding, so that better synergistic enhancement effect is achieved, TGF-beta expression is further promoted, generation of type I and type III collagen is further promoted, and MMP-1 inhibition effect is improved.
The preparation method of the mannitol derivative comprises the following steps: mixing 5-chlorosulfonyl-2-hydroxybenzoic acid with mannitol to react and obtain mannitol derivative.
Further specifically, the method for producing the mannitol derivative comprises:
and adding dry pyridine into 5-chlorosulfonyl-2-hydroxybenzoic acid, stirring for dissolving, slowly adding D-mannitol under ice-water bath, stirring for 2-4 hours, standing overnight, adding 1/3 volume of cold water, stirring for 2-4 hours at normal temperature, adding into ice water containing concentrated hydrochloric acid while stirring, carrying out vacuum filtration, washing with absolute ethanol to neutrality, dehydrating, recrystallizing with absolute ethanol, and carrying out vacuum drying to obtain mannitol derivatives.
Specifically, the solid-to-liquid ratio of 5-chlorosulfonyl-2-hydroxybenzoic acid to pyridine is 0.5-0.7 g:1mL; the molar ratio of the D-mannitol to the 5-chlorosulfonyl-2-hydroxybenzoic acid is 1:6-8; the volume ratio of the ice water to the pyridine is 2.5-3.5:1; the volume ratio of the concentrated hydrochloric acid to the ice water is 1:2-3.
More preferably, a sodium hyaluronate derivative comprises an esterified derivative, the chemical structure of which takes a sodium hyaluronate chain as a main chain structure, and the chemical grafting modification comprises chlorine-containing compounds;
the chlorine-containing compound comprises at least one of N1-gamma-chloropropyl-N4-beta-hydroxyethyl piperazine and 1, 2-bis (2-chloroethoxy) ethane.
Specifically, the chlorine-containing compound is attached to the sodium hyaluronate chain structure by a chemical reaction of chlorine in its structure with carboxyl groups in the sodium hyaluronate structure.
Specifically, the method for producing a hyaluronic acid derivative comprises: mixing N1-gamma-chloropropyl-N4-beta-hydroxyethyl piperazine and/or 1, 2-di (2-chloroethoxy) ethane with sodium hyaluronate, and reacting under the condition of tetrabutylammonium chloride to obtain the hyaluronic acid derivative.
Further specifically, the preparation method of the sodium hyaluronate derivative comprises the following steps:
and (3) taking sodium hyaluronate, N1-gamma-chloropropyl-N4-beta-hydroxyethyl piperazine and 1, 2-bis (2-chloroethoxy) ethane, adding tetrabutylammonium chloride and N, N-dimethyl diamide, stirring and mixing uniformly, heating to 60-70 ℃, stirring and reacting for 2-3 days, recovering N, N-dimethyl diamide under reduced pressure, adding water, regulating the pH to 3-4, precipitating, filtering and vacuum drying to obtain sodium hyaluronate derivatives.
Specifically, the mass ratio of the sodium hyaluronate to the N1-gamma-chloropropyl-N4-beta-hydroxyethyl piperazine is 1:0.1-0.2; the molar ratio of the N1-gamma-chloropropyl-N4-beta-hydroxyethyl piperazine to the 1, 2-di (2-chloroethoxy) ethane is 1:0.4-0.6; the mass ratio of tetrabutyl ammonium chloride to sodium hyaluronate is 1:0.3-0.4.
Specifically, the solid-to-liquid ratio of sodium hyaluronate to N, N-dimethyl diamide is 0.2-0.35 g/1 mL; the volume ratio of water to N, N-dimethyl diamide is 1.4-1.8:1.
The invention also discloses application of the sodium hyaluronate derivative in preparing medical cosmetic filler, cosmetics or skin care products or medicaments.
The invention also discloses application of the composition in preparing medical cosmetic filler, cosmetics or skin care products or medicaments.
An aqueous product comprising the above composition.
Specifically, the concentration of sodium hyaluronate or sodium hyaluronate derivatives in the aqueous solution product is 0.2-1 wt%; preferably 0.5 to 1wt%.
Specifically, the concentration of the collagen pentapeptide in the aqueous solution product is 2-2000 ppm, preferably 20-500 ppm; more preferably 50 to 100ppm.
A powder product comprising the above composition.
Specifically, sodium hyaluronate or sodium hyaluronate crosslinked derivatives in the powder product are dissolved in water when in use, and the concentration is 0.2-1.5wt%.
Specifically, the concentration of the collagen pentapeptide in the powder product is 2-2000 ppm after the collagen pentapeptide is dissolved in water during use.
Compared with the prior art, the invention has the following beneficial effects:
the invention uses the combination of the sodium hyaluronate and the collagen pentapeptide to obtain a composition product, has better effect when being used in combination, generates the synergistic enhancement effect, obviously enhances the collagen regeneration stimulation capability and increases the MMP-1 inhibition effect. Meanwhile, the invention provides a technical scheme of effective proportion of sodium hyaluronate, polypeptide and mannitol, which is favorable for skin rejuvenation and has excellent effect; the mannitol derivative is prepared by adopting the 5-chlorosulfonyl-2-hydroxybenzoic acid modified mannitol, the mannitol derivative has better antioxidation capability, and is added into the composition as an auxiliary component, and each component is compounded for use, so that better synergistic enhancement effect is achieved, the expression of TGF-beta is further promoted, the generation of collagen is further promoted, the MMP-1 inhibition effect is increased, and the collagen regeneration stimulation capability is obviously enhanced.
Therefore, the invention provides a composition of sodium hyaluronate and collagen pentapeptide and application thereof, wherein the sodium hyaluronate and the collagen pentapeptide are combined and applied, so that the composition has good collagen regeneration stimulating capability and synergistic enhancement effect; mannitol or its derivative is added simultaneously to further promote the collagen regeneration stimulating capacity of the composition.
Drawings
FIG. 1 shows the cytotoxicity test results in the examples of the present invention;
FIG. 2 is a graph showing the relative expression of the related anti-aging genes measured after the treatment of samples of collagen pentapeptide aqueous solution with different concentrations according to the embodiment of the present invention;
FIG. 3 is a graph showing the relative expression of the genes involved in determining the relative expression of the genes involved in the present invention after the treatment of aqueous HA solutions of different concentrations;
FIG. 4 is an infrared test result of sodium hyaluronate derivatives and sodium hyaluronate in an example of the present invention;
FIG. 5 shows the results of an infrared test of mannitol derivatives in the examples of the present invention.
Detailed Description
For the purpose of making the objects, technical solutions and advantages of the embodiments of the present invention more apparent, the following describes in detail various embodiments of the present invention with reference to the embodiments. However, those of ordinary skill in the art will understand that in various embodiments of the present invention, numerous technical details have been set forth in order to provide a better understanding of the present application. However, the technical solutions claimed in the present application can be implemented without these technical details and with various changes and modifications based on the following embodiments.
Example 1:
an aqueous product comprising sodium hyaluronate and collagen pentapeptide, formulated in a series of concentrations according to table 1:
TABLE 1 sample concentration ratios
Product cytotoxicity assays
The MTT method is used for testing cytotoxicity, HA is mixed with DMSO solvent containing bovine embryo serum to be used as a liquid culture medium, and the HA concentration in the culture medium simulates the HA concentration in a product.
The test results are shown in FIG. 1. From the analysis in the figure, three concentrations of 0.2%, 0.6% and 1.2% of sodium hyaluronate are selected and respectively added with KTTKS with different concentrations, and the obtained solution has no cytotoxicity reaction.
Example 2:
exploration of synergistic action mechanism of HA+KTTKS
The present study uses the QuantiGene Plex (QGP) method based on the Luminex200 suspension chip multiplex detection system, a detection method that incorporates branched DNA technology that directly measures RNA transcripts by signal amplification. The Luminex200 suspension chip multiplex detection system combines microsphere double fluorescent markers and liquid flow dispersion laser automatic detection technology, and can realize multichannel high-flux analysis of nucleic acid, antibodies and the like.
In this experiment, a total of 5 samples were designed to act on aged human fibroblasts (HFF 50) of 50 years old, as a control group, and the specific list is shown in Table 2 below:
TABLE 2 grouping of samples
Meanwhile, 6 year old human fibroblasts (HFF 6) were provided as an additional verification to approximately determine the regulatory range of the anti-aging gene expression.
Analysis of test results:
FIG. 2 shows the relative expression of the relevant anti-aging gene (TGF-. Beta.as transforming factor-. Beta.; COL1α1 is type I collagen gene) as determined by the QuantiGene Plex (QGP) assay after treatment of 50 year old human fibroblasts with samples of collagen pentapeptides at various concentrations. From the analysis in the figure, the stimulation effect of KTTKS on type I collagen is mainly related to a biosynthesis pathway, and the synthesis of type I collagen is further promoted by promoting the up-regulation of the expression of transforming growth factor beta (TGF-beta); while maintaining mRNA stability during the relevant process.
FIG. 3 shows the relative expression of the relevant anti-aging genes (TGF-. Beta.as the growth factor. Beta.; MMP1 as matrix metalloproteinase 1; COL1α1 as collagen type I gene; COL3α1 as collagen type III gene) as determined by the QuantiGene Plex (QGP) assay after treatment of 50 year old human fibroblasts with different HA samples. From the analysis in the figure, the expression of the growth conversion factor beta (TGF-beta) increases with the increase of the concentration of sodium hyaluronate, and the dose-dependent relationship is shown, while the introduction of KTTKS (100 ppm) has a strong promotion effect on the expression of TGF-beta, and the gain effect of the TGF-beta is not the superposition of the simple two effects. Therefore, HA and KTTKS jointly promote the expression of TGF-beta, further promote the generation of type I and type III collagen, and have synergistic promotion effect between HA and KTTKS.
Meanwhile, KTTKS HAs no obvious effect of inhibiting matrix metalloproteinase (MMP-1), and HAs excellent MMP-1 inhibition effect when combined with HA, thus proving that the KTTKS and the HA have synergistic effect. One target for exogenous aging factors is the dermal extracellular matrix produced by dermal fibroblasts. Ultraviolet light (UVB) activates proteolytic cleavage of dermal collagen fibrils by up-regulating the base MMP-1. As a result of collagen degradation, ECM networks are disrupted despite de novo collagen synthesis and repair, small defects remain in the ECM and accumulate over time. Therefore, the inhibition of MMP-1 can slow down the degradation rate of extracellular matrix and further promote the regeneration of collagen
In conclusion, the HA and the KTTKS are combined and applied for the first time and are subjected to systematic research, so that the HA and the KTTKS have good collagen regeneration stimulation capability, and the effect is better when the HA and the KTTKS are combined.
Example 3:
preparation of sodium hyaluronate crosslinked derivatives
Dissolving sodium hyaluronate (commercially available) with molecular weight of 8-15 ten thousand Da and sodium hyaluronate (commercially available) with molecular weight of 80-200 ten thousand Da in water, stirring and mixing uniformly at normal temperature, heating to 80 ℃, stirring for 1h, adding citric acid to adjust pH to 4.5, transferring into a high-pressure reaction kettle, reacting at 180 ℃ for 2h, cooling to 80 ℃, stirring for 10min, transferring into the high-pressure reaction kettle again, staying at 120 ℃ for 6h, cooling to normal temperature, and forming the partially physically crosslinked sodium hyaluronate 1. In the specific preparation process, the molar ratio of HA with the molecular weight of 8-15 ten thousand Da to HA with the molecular weight of 80-200 ten thousand Da is 3:1, the total concentration of sodium hyaluronate is 1.5wt%.
The preparation of the sodium hyaluronate crosslinked material 2 differs from the sodium hyaluronate crosslinked material 1 in that: in the preparation process, the molecular weight of 80-200 ten thousand Da is changed into 15-80 ten thousand Da (purchased in market);
the preparation of sodium hyaluronate crosslinker 3 differs from that of sodium hyaluronate crosslinker 1 in that: in the preparation process, the molecular weight of 80-200 ten thousand Da is changed into 1-8 ten thousand Da (commercially available).
Example 4:
synergistic effect of series of sodium hyaluronate crosslinked derivatives and KTTKS
The specific test method is the same as in example 2. The test sample M1 was: sodium hyaluronate crosslinker 1 (prepared in example 3) at a concentration of 1.5% + KTTKS100ppm; test sample M2 was: sodium hyaluronate crosslinker 2 (prepared in example 3) at a concentration of 1.5% + KTTKS100ppm; test sample M3 (prepared in example 3) was: sodium hyaluronate cross-linked 3 concentration 1.5% + KTTKS100ppm; test sample M4 was: sodium hyaluronate crosslinker 1 concentration 1.5%; test sample M5 was: sodium hyaluronate crosslinker 2 concentration 1.5%; test sample M6 was: the concentration of the sodium hyaluronate cross-linked material 3 was 1.5%.
The test results are shown in table 3:
TABLE 3 results of relative Gene expression level test
From the analysis of the data in Table 3, it was found that the composition of crosslinked sodium hyaluronate and KTTKS prepared in this example, in which the composition of low crosslinking degree, medium crosslinking degree and high crosslinking degree was obtained according to the change in molecular weight, and that the composition of sodium hyaluronate M4 of high crosslinking degree and the composition of sodium hyaluronate M1 and KTTKS showed the highest relative expression of TGF-. Beta., COL1α1 and COL3α1 genes, showed that sodium hyaluronate for filling effect, which produced appropriate foreign body reaction, was able to stimulate collagen regeneration through TGF-. Beta.signaling pathway. Under the same group of conditions, the introduction of KTTKS can further improve the forward gain effect. The relative expression of MMP1 genes was less different between the same groups, but the expression of MMP1 was reduced and the degree of aging was reduced compared to crosslinked HA alone and compositions containing KTTKS.
Product cytotoxicity assays
The test method was the same as in example 1. From the analysis of the test results, the test sample solution obtained in this example showed no cytotoxic reaction.
Example 5:
synergistic effect of sodium hyaluronate derivative and KTTKS
The specific test method is the same as in example 2. The test sample M1 was: the concentration of the sodium hyaluronate derivative 1 prepared in the example is 0.6% + KTTKS100ppm; test sample M2 was: the concentration of the sodium hyaluronate derivative 2 prepared in the example is 0.6% + KTTKS100ppm; test sample M3 was: the concentration of the sodium hyaluronate derivative 3 prepared in this example was 0.6% + KTTKS100ppm.
Preparation of sodium hyaluronate derivative 1:
sodium hyaluronate (20-40 ten thousand molecular weight, commercially available), N1-gamma-chloropropyl-N4-beta-hydroxyethyl piperazine and 1, 2-di (2-chloroethoxy) ethane are taken, tetrabutylammonium chloride and N, N-dimethyl diamide are added, stirred and mixed uniformly, the temperature is raised to 68 ℃, the stirring reaction is carried out for 2.5 days, the N, N-dimethyl diamide is recovered under reduced pressure, water is added, the pH is regulated to 3.6, precipitation is separated out, and the sodium hyaluronate derivative 1 is obtained through filtration and vacuum drying. In the specific preparation process, the mass ratio of the sodium hyaluronate to the N1-gamma-chloropropyl-N4-beta-hydroxyethyl piperazine is 1:0.16; the molar ratio of N1-gamma-chloropropyl-N4-beta-hydroxyethyl piperazine to 1, 2-bis (2-chloroethoxy) ethane is 1:0.51; the mass ratio of tetrabutyl ammonium chloride to sodium hyaluronate is 1:0.35; the solid-to-liquid ratio of sodium hyaluronate to N, N-dimethylformamide was 0.28g:1mL; the volume ratio of water to N, N-dimethylformamide was 1.6:1.
The preparation of sodium hyaluronate derivative 2 differs from sodium hyaluronate derivative 1 in that: in the preparation process, 1, 2-bis (2-chloroethoxy) ethane is replaced by N1-gamma-chloropropyl-N4-beta-hydroxyethyl piperazine with equal molar weight.
The preparation of sodium hyaluronate derivative 3 differs from sodium hyaluronate derivative 1 in that: in the preparation process, 1, 2-bis (2-chloroethoxy) ethane with equal molar weight is adopted to replace N1-gamma-chloropropyl-N4-beta-hydroxyethyl piperazine.
Analysis of results:
the sodium hyaluronate derivative prepared by the embodiment is used together with KTTKS, the relative gene expression quantity of TGF-beta, COL1 alpha 1 and COL3 alpha 1 is obviously higher than that of sodium hyaluronate and KTTKS, the relative gene expression quantity of MMO1 is obviously lower than that of sodium hyaluronate and KTTKS, the effect of a test sample M1 is obviously better than that of a test sample M2 and a test sample M3, the sodium hyaluronate is subjected to chemical modification by using N1-gamma-chloropropyl-N4-beta-hydroxyethyl piperazine to obtain the sodium hyaluronate derivative, and the sodium hyaluronate derivative is compounded with KTTKS, so that the compounding effect of the sodium hyaluronate derivative can be further enhanced, the expression of TGF-beta is better promoted, the generation of type I collagen and type III collagen is further promoted, and the MMP-1 inhibition effect is better; and under the condition that 1, 2-di (2-chloroethoxy) ethane and N1-gamma-chloropropyl-N4-beta-hydroxyethyl piperazine modified sodium hyaluronate are used simultaneously, the two have synergistic enhancement function, and the collagen regeneration stimulation capability is obviously enhanced.
Product cytotoxicity assays
The test method was the same as in example 1. From the analysis of the test results, the test sample solution obtained in this example showed no cytotoxic reaction.
Example 6:
investigation of the Co-use conditions of HA+KTTKS+mannitol or mannitol derivatives
The specific test method is the same as in example 2. Test sample N1 was: HA concentration 0.6% + KTTKS100ppm; +mannitol 1%; test sample N2 was: HA concentration 0.6% + KTTKS100ppm; +mannitol derivative 1%.
Preparation of mannitol derivatives:
adding dry pyridine into 5-chlorosulfonyl-2-hydroxybenzoic acid according to the solid-to-liquid ratio of 0.6g to 1mL, stirring for dissolving, slowly adding D-mannitol (the molar ratio of the D-mannitol to the 5-chlorosulfonyl-2-hydroxybenzoic acid is 1 to 7.2) under an ice water bath, stirring for 3 hours, standing overnight, adding 1/3 volume of cold water, stirring at normal temperature for 3 hours, adding into ice water containing concentrated hydrochloric acid (the volume ratio of the concentrated hydrochloric acid to the ice water is 1 to 2.5, the volume ratio of the ice water to the pyridine is 3 to 1) while stirring, carrying out vacuum suction filtration, washing with absolute ethanol to be neutral, dehydrating, recrystallizing with absolute ethanol, and carrying out vacuum drying to obtain the mannitol derivative.
The test results are shown in table 4:
TABLE 4 results of relative Gene expression level test
As shown by the data analysis in Table 4, the relative expression of the genes of TGF-beta, COL1 alpha 1 and COL3 alpha 1 of the test samples N1 and N2 is obviously higher than that of HA+KTTKS, the relative expression of the genes of MMO1 is obviously lower than that of sodium hyaluronate+KTTKS, and meanwhile, the effect of the test sample N2 is obviously better than that of the test sample N1, and the mannitol or the derivative thereof is compounded with HA and KTTKS to play a role in enhancing, so that the expression of TGF-beta is further promoted, the generation of I-type and III-type collagen is further promoted, and the MMP-1 inhibition effect is better; and the mannitol derivative is obtained by adopting the 5-chlorosulfonyl-2-hydroxybenzoic acid to chemically modify mannitol, so that the promotion and inhibition effects of the mixed sample can be obviously improved.
Product cytotoxicity assays
The test method was the same as in example 1. From the analysis of the test results, the test sample solution obtained in this example showed no cytotoxic reaction.
Example 6:
mannitol and derivative Performance characterization
Determination of free radical scavenging Effect
After DPPH is dissolved in a small amount of toluene, a solution of 120 mu mol/L is prepared by using 50% ethanol, 1.9mL of the solution is taken, 0.1mL of a test sample with the concentration of 2.0g/L is added, the solution is kept stand for 20min at room temperature, and then the absorbance at 525nm is measured. The clearance was calculated according to the following equation:
clearance/% = (A-A) 1 )/(A-A 0 )×100%
Wherein A is the absorbance of a control system, A 1 To test the absorbance of the sample, A 0 Absorbance was measured for the blank.
Mannitol and the mannitol derivatives prepared in example 4 were subjected to the above test, and the results are shown in table 5:
TABLE 5 free radical scavenging effect test results
From the data analysis in table 5, the clearance rate of the mannitol derivative to the DPPH free radical is obviously higher than that of mannitol, which indicates that the mannitol derivative obtained by chemically modifying mannitol with 5-chlorosulfonyl-2-hydroxybenzoic acid can obviously improve the free radical adsorption clearance capability of the mannitol derivative.
Test example 1:
infrared sign
The testing is carried out by adopting a Fourier infrared spectrometer, and the testing range is 500-4000 cm -1 。
The sodium hyaluronate crosslinked product prepared in example 3 was subjected to the above test, and the characteristic peaks in the infrared spectrum thereof were 3275 to 3390 (b), 1615(s), 1405 (m), 1150, 1077, 1045(s), 946 (m), 893 (w). The wave number error of the actually measured band of the fingerprint area is less than +/-5 cm -1 。
The sodium hyaluronate derivative 1 prepared in example 4 and sodium hyaluronate were subjected to the above-described test, and the results are shown in fig. 4. As can be seen from the analysis of the figure, compared with the infrared test result of sodium hyaluronate, in the infrared spectrum of the sodium hyaluronate derivative 1 prepared in example 3, the infrared spectrum is 3000-2800 cm -1 The characteristic absorption peak intensity of methylene in the range is obviously improved, and the peak intensity is 1350-1250cm -1 The characteristic absorption peak intensity of C-N bond in the range is enhanced, and the characteristic absorption peak intensity is 1100-1000 cm -1 The characteristic absorption peak intensity of the C-O bond in the range was enhanced, and the above results indicate that sodium hyaluronate derivative 1 in example 3 was successfully produced.
The mannitol derivatives prepared in example 5 were subjected to the above test, and the results are shown in fig. 5. From the analysis of the graph, 3000-2800 cm -1 Characteristic absorption peak of methyl and methylene in the range of 1500-1400 cm -1 Characteristic absorption peak of benzene ring appeared in the range of 1353cm -1 、1183cm -1 The characteristic absorption peak of s=o appearing nearby, the above result indicates successful preparation of mannitol derivative in example 3.
The conventional technology in the above embodiments is known to those skilled in the art, and thus is not described in detail herein.
The foregoing is merely illustrative of the present invention, and the present invention is not limited thereto, and any person skilled in the art will readily recognize that variations or substitutions are within the scope of the present invention. Therefore, the protection scope of the present invention shall be subject to the protection scope of the claims.
Claims (7)
1. A composition comprising collagen pentapeptide, sodium hyaluronate, and a mannitol derivative comprising the product of 5-chlorosulfonyl-2-hydroxybenzoic acid modified mannitol; or alternatively, the first and second heat exchangers may be,
comprises collagen pentapeptide and sodium hyaluronate derivative, wherein the sodium hyaluronate derivative comprises sodium hyaluronate cross-linked derivative or sodium hyaluronate derivative 1 or sodium hyaluronate derivative 2 or sodium hyaluronate derivative 3;
the sodium hyaluronate crosslinking derivative comprises a mixture crosslinking substance of one or more of sodium hyaluronate with single-chain molecular weight of 1-8 ten thousand, 8-15 ten thousand, 15-30 ten thousand, 30-80 ten thousand and 80-200 ten thousand, and the crosslinking mode of the sodium hyaluronate crosslinking derivative adopts physical crosslinking;
in the preparation of the sodium hyaluronate derivative 1, N1-gamma-chloropropyl-N4-beta-hydroxyethyl piperazine and 1, 2-di (2-chloroethoxy) ethane are mixed with sodium hyaluronate, and the mixture is reacted under the condition of tetrabutylammonium chloride to prepare the hyaluronic acid derivative 1; in the preparation of the sodium hyaluronate derivative 2, N1-gamma-chloropropyl-N4-beta-hydroxyethyl piperazine is mixed with sodium hyaluronate, and the mixture is reacted under the condition of tetrabutylammonium chloride to prepare the sodium hyaluronate derivative 2; in the preparation of the sodium hyaluronate derivative 3, 1, 2-di (2-chloroethoxy) ethane is mixed with sodium hyaluronate and reacts under the condition of tetrabutylammonium chloride to prepare the sodium hyaluronate derivative 3;
the mass ratio of the sodium hyaluronate to the N1-gamma-chloropropyl-N4-beta-hydroxyethyl piperazine is 1:0.1-0.2; the molar ratio of the N1-gamma-chloropropyl-N4-beta-hydroxyethyl piperazine to the 1, 2-di (2-chloroethoxy) ethane is 1:0.4-0.6; the mass ratio of tetrabutyl ammonium chloride to sodium hyaluronate is 1:0.3-0.4;
the collagen pentapeptide is KTTKS, and the KTTKS is lysine-threonine-lysine-serine.
2. The composition of claim 1, wherein: the composition also comprises auxiliary ingredients including one or more of sorbitol and dextran.
3. Use of the composition of claim 1 for the preparation of a medical cosmetic injection filler, a cosmetic or skin care product or medicament.
4. An aqueous product comprising the composition of any one of claims 1-2.
5. The aqueous product of claim 4, wherein: the concentration of the sodium hyaluronate or sodium hyaluronate derivative in the aqueous solution product is 0.2-1.5wt%; the concentration of the collagen pentapeptide is 2-2000 ppm.
6. A powder product comprising the composition of any one of claims 1-2.
7. A powder product as claimed in claim 6, wherein: sodium hyaluronate or sodium hyaluronate crosslinked derivatives in the product are dissolved in water when in use, and the concentration is 0.2-1.5wt%; the concentration of the collagen pentapeptide in the product is 2-2000 ppm after the collagen pentapeptide is dissolved in water during use.
Priority Applications (1)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
CN202311606423.3A CN117304368B (en) | 2023-11-29 | 2023-11-29 | Composition of sodium hyaluronate and collagen pentapeptide and application thereof |
Applications Claiming Priority (1)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
CN202311606423.3A CN117304368B (en) | 2023-11-29 | 2023-11-29 | Composition of sodium hyaluronate and collagen pentapeptide and application thereof |
Publications (2)
Publication Number | Publication Date |
---|---|
CN117304368A CN117304368A (en) | 2023-12-29 |
CN117304368B true CN117304368B (en) | 2024-03-01 |
Family
ID=89285040
Family Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
CN202311606423.3A Active CN117304368B (en) | 2023-11-29 | 2023-11-29 | Composition of sodium hyaluronate and collagen pentapeptide and application thereof |
Country Status (1)
Country | Link |
---|---|
CN (1) | CN117304368B (en) |
Citations (7)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US6140312A (en) * | 1992-02-20 | 2000-10-31 | Hyal Pharmaceutical Corporation | Formulations containing hyaluronic acid |
WO2013028904A2 (en) * | 2011-08-25 | 2013-02-28 | Allergan, Inc. | Dermal filler compositions including antioxidants |
CN109833266A (en) * | 2019-03-29 | 2019-06-04 | 深圳市富可迪生物科技有限公司 | Cosmetic composition |
WO2021018950A1 (en) * | 2019-07-29 | 2021-02-04 | Giuliani S.P.A. | Cosmetic composition with combined filler and skin regenerative effect |
CN115350113A (en) * | 2022-07-04 | 2022-11-18 | 广州环亚化妆品科技股份有限公司 | Composition containing peptide and application thereof |
CN115636884A (en) * | 2022-12-23 | 2023-01-24 | 北京格林意锐医药科技有限责任公司 | Preparation method of sodium hyaluronate derivative, cross-linked sodium hyaluronate and application |
WO2023042119A2 (en) * | 2021-09-16 | 2023-03-23 | Professional Derma Sa | Composition for cosmetic use |
Family Cites Families (3)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US9119974B2 (en) * | 2011-03-04 | 2015-09-01 | Ahmed H. Al-Qahtani | Skin cream |
CZ304512B6 (en) * | 2012-08-08 | 2014-06-11 | Contipro Biotech S.R.O. | Hyaluronic acid derivative, process for its preparation, modification process and use thereof |
US20230240964A1 (en) * | 2021-12-27 | 2023-08-03 | Elastintech, Inc. | Skin-firming cosmetic compositions |
-
2023
- 2023-11-29 CN CN202311606423.3A patent/CN117304368B/en active Active
Patent Citations (7)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US6140312A (en) * | 1992-02-20 | 2000-10-31 | Hyal Pharmaceutical Corporation | Formulations containing hyaluronic acid |
WO2013028904A2 (en) * | 2011-08-25 | 2013-02-28 | Allergan, Inc. | Dermal filler compositions including antioxidants |
CN109833266A (en) * | 2019-03-29 | 2019-06-04 | 深圳市富可迪生物科技有限公司 | Cosmetic composition |
WO2021018950A1 (en) * | 2019-07-29 | 2021-02-04 | Giuliani S.P.A. | Cosmetic composition with combined filler and skin regenerative effect |
WO2023042119A2 (en) * | 2021-09-16 | 2023-03-23 | Professional Derma Sa | Composition for cosmetic use |
CN115350113A (en) * | 2022-07-04 | 2022-11-18 | 广州环亚化妆品科技股份有限公司 | Composition containing peptide and application thereof |
CN115636884A (en) * | 2022-12-23 | 2023-01-24 | 北京格林意锐医药科技有限责任公司 | Preparation method of sodium hyaluronate derivative, cross-linked sodium hyaluronate and application |
Non-Patent Citations (3)
Title |
---|
何秋星.《化妆品制剂学》.中国医药科学技术出版社,2021,第150-152页. * |
新型透明质酸钠凝胶的制备及性能研究;贾柯;《中国优秀硕士学位论文全文数据库 医药卫生科技辑》(第03期);第21页第3.2.2节 * |
载川芎嗪透明质酸水凝胶调控巨噬细胞促进糖尿病伤口愈合的研究;初迪;《万方学位论文》;第37页第3.3.1节、第41页第3.3.5节 * |
Also Published As
Publication number | Publication date |
---|---|
CN117304368A (en) | 2023-12-29 |
Similar Documents
Publication | Publication Date | Title |
---|---|---|
JP7274783B2 (en) | Reduced and oxidized polysaccharides and methods of their use | |
Zhang et al. | Hydrogels based on pH-responsive reversible carbon–nitrogen double-bond linkages for biomedical applications | |
Zhu et al. | Incorporation of ROS-responsive substance P-loaded zeolite imidazolate framework-8 nanoparticles into a Ca2+-cross-linked alginate/pectin hydrogel for wound dressing applications | |
EP2772273B9 (en) | Viscoelastic gels as novel fillers | |
JPH02504163A (en) | cross-linked carboxypolysaccharide | |
EP2222713B1 (en) | Mixed butyric-formic esters of acid polysaccharides, and their preparation and use as skin cosmetics | |
CN114869796A (en) | Whitening and skin-brightening repair composition containing active polypeptide and application thereof | |
CN115887742B (en) | Preparation method of antibacterial functional collagen-based injectable self-repairing hydrogel | |
CN117304368B (en) | Composition of sodium hyaluronate and collagen pentapeptide and application thereof | |
Cao et al. | Starch and chitosan-based antibacterial dressing for infected wound treatment via self-activated NO release strategy | |
CN114478829B (en) | Hyaluronic acid crosslinked active material composition, preparation method and application | |
CN113876788B (en) | Rhodiola rosea glycoside hydrogel preparation and preparation method and application thereof | |
CN113827501B (en) | Hyaluronic acid hydrogel mask with skin repair function and preparation method thereof | |
CN115778845A (en) | Protein composition and preparation method and application thereof | |
EP2178908B1 (en) | Peptides and pharmaceutical compositions for treating connective tissue | |
CN105199115B (en) | Preparation and application of injectable hydrogel with porous three-dimensional network structure | |
KR102132478B1 (en) | Hyaluronic Acid-Polydeoxyribonucleotide Complex and Its Film and Preparation Method Thereof | |
CN114350035B (en) | Hyaluronic acid crosslinking active material and application thereof | |
Liang et al. | A novel targeting drug delivery system based on self-assembled peptide hydrogel | |
CN114621323B (en) | Polypeptide compound with skin repairing effect and preparation method and application thereof | |
CN112876535B (en) | Bioactive polypeptides and uses thereof | |
JP6578323B2 (en) | Filaggrin production promoter | |
CN113145030A (en) | Supramolecular hydrogel and preparation method thereof | |
Jawarkar et al. | NATURAL-BASED HYDROGELS IN COSMETICS: A COMPREHENSIVE REVIEW | |
CN116327610A (en) | Botulinum-like polypeptide nano micelle and preparation method and application thereof |
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 |