CN115584037A - Crosslinked gel material containing polymer microspheres, preparation method thereof and injection filler - Google Patents
Crosslinked gel material containing polymer microspheres, preparation method thereof and injection filler Download PDFInfo
- Publication number
- CN115584037A CN115584037A CN202211280983.XA CN202211280983A CN115584037A CN 115584037 A CN115584037 A CN 115584037A CN 202211280983 A CN202211280983 A CN 202211280983A CN 115584037 A CN115584037 A CN 115584037A
- Authority
- CN
- China
- Prior art keywords
- microspheres
- hyaluronic acid
- cross
- water
- gel
- 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.)
- Pending
Links
- 239000004005 microsphere Substances 0.000 title claims abstract description 92
- 239000000945 filler Substances 0.000 title claims abstract description 40
- 229920000642 polymer Polymers 0.000 title claims abstract description 34
- 239000000463 material Substances 0.000 title claims abstract description 30
- 238000002347 injection Methods 0.000 title claims abstract description 26
- 239000007924 injection Substances 0.000 title claims abstract description 26
- 238000002360 preparation method Methods 0.000 title claims abstract description 19
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims abstract description 67
- 230000001954 sterilising effect Effects 0.000 claims abstract description 43
- 238000000034 method Methods 0.000 claims abstract description 41
- 239000003431 cross linking reagent Substances 0.000 claims abstract description 30
- 238000004132 cross linking Methods 0.000 claims abstract description 23
- 238000002156 mixing Methods 0.000 claims abstract description 15
- 239000003960 organic solvent Substances 0.000 claims abstract description 15
- 229920003232 aliphatic polyester Polymers 0.000 claims abstract description 13
- 230000001804 emulsifying effect Effects 0.000 claims abstract description 12
- 239000004094 surface-active agent Substances 0.000 claims abstract description 11
- 239000000126 substance Substances 0.000 claims abstract description 10
- 229920000249 biocompatible polymer Polymers 0.000 claims abstract description 9
- 239000000839 emulsion Substances 0.000 claims abstract description 8
- 239000007764 o/w emulsion Substances 0.000 claims abstract description 7
- 229920002521 macromolecule Polymers 0.000 claims abstract description 6
- 239000002904 solvent Substances 0.000 claims abstract description 6
- 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 109
- 229920002674 hyaluronan Polymers 0.000 claims description 109
- 229960003160 hyaluronic acid Drugs 0.000 claims description 109
- 238000003756 stirring Methods 0.000 claims description 44
- 238000000502 dialysis Methods 0.000 claims description 41
- 238000004659 sterilization and disinfection Methods 0.000 claims description 40
- 239000004632 polycaprolactone Substances 0.000 claims description 32
- 229920001610 polycaprolactone Polymers 0.000 claims description 32
- YMWUJEATGCHHMB-UHFFFAOYSA-N Dichloromethane Chemical compound ClCCl YMWUJEATGCHHMB-UHFFFAOYSA-N 0.000 claims description 30
- 238000011049 filling Methods 0.000 claims description 25
- 239000000843 powder Substances 0.000 claims description 25
- 239000000243 solution Substances 0.000 claims description 22
- 239000013067 intermediate product Substances 0.000 claims description 20
- 230000008961 swelling Effects 0.000 claims description 15
- 239000008215 water for injection Substances 0.000 claims description 15
- 238000010438 heat treatment Methods 0.000 claims description 13
- XEKOWRVHYACXOJ-UHFFFAOYSA-N Ethyl acetate Chemical compound CCOC(C)=O XEKOWRVHYACXOJ-UHFFFAOYSA-N 0.000 claims description 12
- 239000004372 Polyvinyl alcohol Substances 0.000 claims description 11
- 238000001723 curing Methods 0.000 claims description 11
- 229920000166 polytrimethylene carbonate Polymers 0.000 claims description 11
- 229920002451 polyvinyl alcohol Polymers 0.000 claims description 11
- 239000007853 buffer solution Substances 0.000 claims description 10
- -1 polytrimethylene carbonate Polymers 0.000 claims description 10
- 238000007873 sieving Methods 0.000 claims description 10
- 238000005406 washing Methods 0.000 claims description 9
- 108010035532 Collagen Chemical class 0.000 claims description 8
- 102000008186 Collagen Human genes 0.000 claims description 8
- 238000010926 purge Methods 0.000 claims description 8
- 229920001436 collagen Chemical class 0.000 claims description 7
- ZWEHNKRNPOVVGH-UHFFFAOYSA-N 2-Butanone Chemical compound CCC(C)=O ZWEHNKRNPOVVGH-UHFFFAOYSA-N 0.000 claims description 6
- WYURNTSHIVDZCO-UHFFFAOYSA-N Tetrahydrofuran Chemical compound C1CCOC1 WYURNTSHIVDZCO-UHFFFAOYSA-N 0.000 claims description 6
- YXFVVABEGXRONW-UHFFFAOYSA-N Toluene Chemical compound CC1=CC=CC=C1 YXFVVABEGXRONW-UHFFFAOYSA-N 0.000 claims description 6
- 230000005855 radiation Effects 0.000 claims description 6
- CSCPPACGZOOCGX-UHFFFAOYSA-N Acetone Chemical compound CC(C)=O CSCPPACGZOOCGX-UHFFFAOYSA-N 0.000 claims description 4
- LZCLXQDLBQLTDK-UHFFFAOYSA-N ethyl 2-hydroxypropanoate Chemical compound CCOC(=O)C(C)O LZCLXQDLBQLTDK-UHFFFAOYSA-N 0.000 claims description 4
- 229930195733 hydrocarbon Natural products 0.000 claims description 4
- 150000002430 hydrocarbons Chemical class 0.000 claims description 4
- 229920000747 poly(lactic acid) Polymers 0.000 claims description 4
- SECXISVLQFMRJM-UHFFFAOYSA-N N-Methylpyrrolidone Chemical compound CN1CCCC1=O SECXISVLQFMRJM-UHFFFAOYSA-N 0.000 claims description 3
- 239000012298 atmosphere Substances 0.000 claims description 3
- 230000001681 protective effect Effects 0.000 claims description 3
- 150000003384 small molecules Chemical class 0.000 claims description 3
- YLQBMQCUIZJEEH-UHFFFAOYSA-N tetrahydrofuran Natural products C=1C=COC=1 YLQBMQCUIZJEEH-UHFFFAOYSA-N 0.000 claims description 3
- 229920002134 Carboxymethyl cellulose Chemical class 0.000 claims description 2
- 229920001661 Chitosan Chemical class 0.000 claims description 2
- HEDRZPFGACZZDS-UHFFFAOYSA-N Chloroform Chemical compound ClC(Cl)Cl HEDRZPFGACZZDS-UHFFFAOYSA-N 0.000 claims description 2
- 239000004971 Cross linker Substances 0.000 claims description 2
- 229920002307 Dextran Polymers 0.000 claims description 2
- AEMRFAOFKBGASW-UHFFFAOYSA-N Glycolic acid Polymers OCC(O)=O AEMRFAOFKBGASW-UHFFFAOYSA-N 0.000 claims description 2
- 229920000954 Polyglycolide Polymers 0.000 claims description 2
- 229920002472 Starch Polymers 0.000 claims description 2
- 239000002253 acid Substances 0.000 claims description 2
- 239000001768 carboxy methyl cellulose Chemical class 0.000 claims description 2
- 235000010948 carboxy methyl cellulose Nutrition 0.000 claims description 2
- 239000008112 carboxymethyl-cellulose Chemical class 0.000 claims description 2
- 229960001701 chloroform Drugs 0.000 claims description 2
- 229920001577 copolymer Polymers 0.000 claims description 2
- 229940116333 ethyl lactate Drugs 0.000 claims description 2
- 150000002576 ketones Chemical class 0.000 claims description 2
- 239000000178 monomer Substances 0.000 claims description 2
- 229920002463 poly(p-dioxanone) polymer Polymers 0.000 claims description 2
- 239000000622 polydioxanone Substances 0.000 claims description 2
- 229920000136 polysorbate Polymers 0.000 claims description 2
- 239000008107 starch Substances 0.000 claims description 2
- 235000019698 starch Nutrition 0.000 claims description 2
- 239000000718 radiation-protective agent Substances 0.000 claims 1
- 238000004519 manufacturing process Methods 0.000 abstract description 15
- 239000012071 phase Substances 0.000 description 54
- 229920000728 polyester Polymers 0.000 description 34
- HEMHJVSKTPXQMS-UHFFFAOYSA-M Sodium hydroxide Chemical compound [OH-].[Na+] HEMHJVSKTPXQMS-UHFFFAOYSA-M 0.000 description 27
- 230000008569 process Effects 0.000 description 21
- 239000000047 product Substances 0.000 description 19
- 238000006243 chemical reaction Methods 0.000 description 17
- 238000005303 weighing Methods 0.000 description 17
- HPILSDOMLLYBQF-UHFFFAOYSA-N 2-[1-(oxiran-2-ylmethoxy)butoxymethyl]oxirane Chemical compound C1OC1COC(CCC)OCC1CO1 HPILSDOMLLYBQF-UHFFFAOYSA-N 0.000 description 15
- 229920001432 poly(L-lactide) Polymers 0.000 description 15
- NBIIXXVUZAFLBC-UHFFFAOYSA-N Phosphoric acid Chemical compound OP(O)(O)=O NBIIXXVUZAFLBC-UHFFFAOYSA-N 0.000 description 14
- 238000001035 drying Methods 0.000 description 14
- 238000004321 preservation Methods 0.000 description 14
- 238000004945 emulsification Methods 0.000 description 12
- 238000004108 freeze drying Methods 0.000 description 11
- IJGRMHOSHXDMSA-UHFFFAOYSA-N Atomic nitrogen Chemical compound N#N IJGRMHOSHXDMSA-UHFFFAOYSA-N 0.000 description 10
- 230000000694 effects Effects 0.000 description 9
- 239000007788 liquid Substances 0.000 description 9
- 239000000203 mixture Substances 0.000 description 9
- 229910000147 aluminium phosphate Inorganic materials 0.000 description 7
- 230000015556 catabolic process Effects 0.000 description 6
- 238000006731 degradation reaction Methods 0.000 description 6
- 238000001727 in vivo Methods 0.000 description 6
- 230000001788 irregular Effects 0.000 description 6
- 238000000746 purification Methods 0.000 description 6
- 238000001816 cooling Methods 0.000 description 5
- 238000005520 cutting process Methods 0.000 description 5
- 238000010586 diagram Methods 0.000 description 5
- 238000004090 dissolution Methods 0.000 description 5
- 230000002500 effect on skin Effects 0.000 description 5
- 229910052757 nitrogen Inorganic materials 0.000 description 5
- 239000002245 particle Substances 0.000 description 5
- 210000001519 tissue Anatomy 0.000 description 5
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 description 4
- 229920002385 Sodium hyaluronate Polymers 0.000 description 4
- 230000009286 beneficial effect Effects 0.000 description 4
- 229940010747 sodium hyaluronate Drugs 0.000 description 4
- 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 compound [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 description 4
- 238000000859 sublimation Methods 0.000 description 4
- 230000008022 sublimation Effects 0.000 description 4
- 238000012360 testing method Methods 0.000 description 4
- 239000008346 aqueous phase Substances 0.000 description 3
- 230000007547 defect Effects 0.000 description 3
- 238000001125 extrusion Methods 0.000 description 3
- 238000012986 modification Methods 0.000 description 3
- 230000004048 modification Effects 0.000 description 3
- 239000002994 raw material Substances 0.000 description 3
- UXVMQQNJUSDDNG-UHFFFAOYSA-L Calcium chloride Chemical compound [Cl-].[Cl-].[Ca+2] UXVMQQNJUSDDNG-UHFFFAOYSA-L 0.000 description 2
- 229920001634 Copolyester Polymers 0.000 description 2
- 201000004624 Dermatitis Diseases 0.000 description 2
- 241001465754 Metazoa Species 0.000 description 2
- CDBYLPFSWZWCQE-UHFFFAOYSA-L Sodium Carbonate Chemical compound [Na+].[Na+].[O-]C([O-])=O CDBYLPFSWZWCQE-UHFFFAOYSA-L 0.000 description 2
- 230000009471 action Effects 0.000 description 2
- ZRBROGSAUIUIJE-UHFFFAOYSA-N azanium;azane;chloride Chemical compound N.[NH4+].[Cl-] ZRBROGSAUIUIJE-UHFFFAOYSA-N 0.000 description 2
- WERYXYBDKMZEQL-UHFFFAOYSA-N butane-1,4-diol Chemical compound OCCCCO WERYXYBDKMZEQL-UHFFFAOYSA-N 0.000 description 2
- 239000001110 calcium chloride Substances 0.000 description 2
- 229910001628 calcium chloride Inorganic materials 0.000 description 2
- 239000003795 chemical substances by application Substances 0.000 description 2
- 238000002316 cosmetic surgery Methods 0.000 description 2
- 239000003814 drug Substances 0.000 description 2
- 238000010828 elution Methods 0.000 description 2
- 239000012467 final product Substances 0.000 description 2
- 238000004806 packaging method and process Methods 0.000 description 2
- 239000004626 polylactic acid Substances 0.000 description 2
- 229940071643 prefilled syringe Drugs 0.000 description 2
- 230000035484 reaction time Effects 0.000 description 2
- 150000003839 salts Chemical class 0.000 description 2
- 238000007789 sealing Methods 0.000 description 2
- 238000003860 storage Methods 0.000 description 2
- 230000037303 wrinkles Effects 0.000 description 2
- SHKUUQIDMUMQQK-UHFFFAOYSA-N 2-[4-(oxiran-2-ylmethoxy)butoxymethyl]oxirane Chemical compound C1OC1COCCCCOCC1CO1 SHKUUQIDMUMQQK-UHFFFAOYSA-N 0.000 description 1
- KZDCPJDWDXMJRA-UHFFFAOYSA-N 2-hydroxybutanoic acid;pentanoic acid Chemical compound CCCCC(O)=O.CCC(O)C(O)=O KZDCPJDWDXMJRA-UHFFFAOYSA-N 0.000 description 1
- AFENDNXGAFYKQO-UHFFFAOYSA-N 2-hydroxybutyric acid Chemical compound CCC(O)C(O)=O AFENDNXGAFYKQO-UHFFFAOYSA-N 0.000 description 1
- SJZRECIVHVDYJC-UHFFFAOYSA-M 4-hydroxybutyrate Chemical compound OCCCC([O-])=O SJZRECIVHVDYJC-UHFFFAOYSA-M 0.000 description 1
- JJTUDXZGHPGLLC-IMJSIDKUSA-N 4511-42-6 Chemical compound C[C@@H]1OC(=O)[C@H](C)OC1=O JJTUDXZGHPGLLC-IMJSIDKUSA-N 0.000 description 1
- 208000005443 Circulating Neoplastic Cells Diseases 0.000 description 1
- 239000004593 Epoxy Substances 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
- 229920002683 Glycosaminoglycan Polymers 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
- MBLBDJOUHNCFQT-LXGUWJNJSA-N N-acetylglucosamine Natural products CC(=O)N[C@@H](C=O)[C@@H](O)[C@H](O)[C@H](O)CO MBLBDJOUHNCFQT-LXGUWJNJSA-N 0.000 description 1
- 229910019142 PO4 Inorganic materials 0.000 description 1
- 229920000331 Polyhydroxybutyrate Polymers 0.000 description 1
- KWYUFKZDYYNOTN-UHFFFAOYSA-M Potassium hydroxide Chemical compound [OH-].[K+] KWYUFKZDYYNOTN-UHFFFAOYSA-M 0.000 description 1
- 208000003251 Pruritus Diseases 0.000 description 1
- 229920001954 Restylane Polymers 0.000 description 1
- 206010045168 Tumour embolism Diseases 0.000 description 1
- 206010046543 Urinary incontinence Diseases 0.000 description 1
- 238000010521 absorption reaction Methods 0.000 description 1
- 238000004026 adhesive bonding Methods 0.000 description 1
- 238000007605 air drying Methods 0.000 description 1
- 150000001299 aldehydes Chemical class 0.000 description 1
- 150000001338 aliphatic hydrocarbons Chemical class 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
- 229920006125 amorphous polymer Polymers 0.000 description 1
- 150000004945 aromatic hydrocarbons Chemical class 0.000 description 1
- 230000033228 biological regulation Effects 0.000 description 1
- 230000036760 body temperature Effects 0.000 description 1
- 210000000988 bone and bone Anatomy 0.000 description 1
- 239000000872 buffer Substances 0.000 description 1
- 239000000501 collagen implant Substances 0.000 description 1
- 150000001875 compounds Chemical class 0.000 description 1
- 238000013270 controlled release Methods 0.000 description 1
- 238000005034 decoration Methods 0.000 description 1
- 230000007423 decrease Effects 0.000 description 1
- 230000003247 decreasing effect Effects 0.000 description 1
- 238000011161 development Methods 0.000 description 1
- 230000018109 developmental process Effects 0.000 description 1
- 150000002016 disaccharides Chemical class 0.000 description 1
- 239000006185 dispersion Substances 0.000 description 1
- 229940079593 drug Drugs 0.000 description 1
- 229920001971 elastomer Polymers 0.000 description 1
- 239000000806 elastomer Substances 0.000 description 1
- 238000010894 electron beam technology Methods 0.000 description 1
- 238000005516 engineering process Methods 0.000 description 1
- RTZKZFJDLAIYFH-UHFFFAOYSA-N ether Substances CCOCC RTZKZFJDLAIYFH-UHFFFAOYSA-N 0.000 description 1
- 230000001815 facial effect Effects 0.000 description 1
- 230000002349 favourable effect Effects 0.000 description 1
- 210000002950 fibroblast Anatomy 0.000 description 1
- 239000004088 foaming agent Substances 0.000 description 1
- 238000007710 freezing Methods 0.000 description 1
- 230000008014 freezing Effects 0.000 description 1
- 238000004388 gamma ray sterilization Methods 0.000 description 1
- 239000007863 gel particle Substances 0.000 description 1
- 230000014509 gene expression Effects 0.000 description 1
- 238000005469 granulation Methods 0.000 description 1
- 230000003179 granulation Effects 0.000 description 1
- 229910052736 halogen Inorganic materials 0.000 description 1
- 125000005843 halogen group Chemical group 0.000 description 1
- FUZZWVXGSFPDMH-UHFFFAOYSA-M hexanoate Chemical compound CCCCCC([O-])=O FUZZWVXGSFPDMH-UHFFFAOYSA-M 0.000 description 1
- 230000007062 hydrolysis Effects 0.000 description 1
- 238000006460 hydrolysis reaction Methods 0.000 description 1
- 206010020718 hyperplasia Diseases 0.000 description 1
- 125000000879 imine group Chemical group 0.000 description 1
- 238000011534 incubation Methods 0.000 description 1
- 208000015181 infectious disease Diseases 0.000 description 1
- 230000036512 infertility Effects 0.000 description 1
- 239000000314 lubricant Substances 0.000 description 1
- 230000014759 maintenance of location Effects 0.000 description 1
- 238000005259 measurement Methods 0.000 description 1
- 230000007246 mechanism Effects 0.000 description 1
- 238000002844 melting Methods 0.000 description 1
- 230000008018 melting Effects 0.000 description 1
- 239000011859 microparticle Substances 0.000 description 1
- 229950006780 n-acetylglucosamine Drugs 0.000 description 1
- 230000000399 orthopedic effect Effects 0.000 description 1
- 230000003204 osmotic effect Effects 0.000 description 1
- 230000000737 periodic effect Effects 0.000 description 1
- NBIIXXVUZAFLBC-UHFFFAOYSA-K phosphate Chemical compound [O-]P([O-])([O-])=O NBIIXXVUZAFLBC-UHFFFAOYSA-K 0.000 description 1
- 239000010452 phosphate Substances 0.000 description 1
- 239000008055 phosphate buffer solution Substances 0.000 description 1
- 229920000071 poly(4-hydroxybutyrate) Polymers 0.000 description 1
- 239000005014 poly(hydroxyalkanoate) Substances 0.000 description 1
- 239000005015 poly(hydroxybutyrate) Substances 0.000 description 1
- 229920000070 poly-3-hydroxybutyrate Polymers 0.000 description 1
- 229920000903 polyhydroxyalkanoate Polymers 0.000 description 1
- 238000005086 pumping Methods 0.000 description 1
- 230000008929 regeneration Effects 0.000 description 1
- 238000011069 regeneration method Methods 0.000 description 1
- 230000008439 repair process Effects 0.000 description 1
- 230000028327 secretion Effects 0.000 description 1
- 230000037339 smooth wrinkles Effects 0.000 description 1
- 239000011734 sodium Substances 0.000 description 1
- 229910000029 sodium carbonate Inorganic materials 0.000 description 1
- 238000007711 solidification Methods 0.000 description 1
- 230000008023 solidification Effects 0.000 description 1
- 230000004936 stimulating effect Effects 0.000 description 1
- 230000000638 stimulation Effects 0.000 description 1
- 238000010254 subcutaneous injection Methods 0.000 description 1
- 239000007929 subcutaneous injection Substances 0.000 description 1
- 150000003457 sulfones Chemical class 0.000 description 1
- 238000001356 surgical procedure Methods 0.000 description 1
- 238000009210 therapy by ultrasound Methods 0.000 description 1
- 231100000331 toxic Toxicity 0.000 description 1
- 230000002588 toxic effect Effects 0.000 description 1
- 238000012546 transfer Methods 0.000 description 1
- 210000001260 vocal cord Anatomy 0.000 description 1
Images
Classifications
-
- C—CHEMISTRY; METALLURGY
- C08—ORGANIC MACROMOLECULAR COMPOUNDS; THEIR PREPARATION OR CHEMICAL WORKING-UP; COMPOSITIONS BASED THEREON
- C08J—WORKING-UP; GENERAL PROCESSES OF COMPOUNDING; AFTER-TREATMENT NOT COVERED BY SUBCLASSES C08B, C08C, C08F, C08G or C08H
- C08J3/00—Processes of treating or compounding macromolecular substances
- C08J3/24—Crosslinking, e.g. vulcanising, of macromolecules
-
- 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
- A61L27/16—Macromolecular materials obtained by reactions only involving carbon-to-carbon unsaturated bonds
-
- 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
- A61L27/18—Macromolecular materials obtained otherwise than by reactions only involving carbon-to-carbon unsaturated bonds
-
- 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
- A61L27/20—Polysaccharides
-
- 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
- A61L27/50—Materials characterised by their function or physical properties, e.g. injectable or lubricating compositions, shape-memory materials, surface modified materials
-
- 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
- A61L27/50—Materials characterised by their function or physical properties, e.g. injectable or lubricating compositions, shape-memory materials, surface modified materials
- A61L27/52—Hydrogels or hydrocolloids
-
- 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
- A61L27/50—Materials characterised by their function or physical properties, e.g. injectable or lubricating compositions, shape-memory materials, surface modified materials
- A61L27/58—Materials at least partially resorbable by the body
-
- C—CHEMISTRY; METALLURGY
- C08—ORGANIC MACROMOLECULAR COMPOUNDS; THEIR PREPARATION OR CHEMICAL WORKING-UP; COMPOSITIONS BASED THEREON
- C08J—WORKING-UP; GENERAL PROCESSES OF COMPOUNDING; AFTER-TREATMENT NOT COVERED BY SUBCLASSES C08B, C08C, C08F, C08G or C08H
- C08J3/00—Processes of treating or compounding macromolecular substances
- C08J3/12—Powdering or granulating
-
- C—CHEMISTRY; METALLURGY
- C08—ORGANIC MACROMOLECULAR COMPOUNDS; THEIR PREPARATION OR CHEMICAL WORKING-UP; COMPOSITIONS BASED THEREON
- C08J—WORKING-UP; GENERAL PROCESSES OF COMPOUNDING; AFTER-TREATMENT NOT COVERED BY SUBCLASSES C08B, C08C, C08F, C08G or C08H
- C08J7/00—Chemical treatment or coating of shaped articles made of macromolecular substances
- C08J7/02—Chemical treatment or coating of shaped articles made of macromolecular substances with solvents, e.g. swelling agents
-
- 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
-
- C—CHEMISTRY; METALLURGY
- C08—ORGANIC MACROMOLECULAR COMPOUNDS; THEIR PREPARATION OR CHEMICAL WORKING-UP; COMPOSITIONS BASED THEREON
- C08J—WORKING-UP; GENERAL PROCESSES OF COMPOUNDING; AFTER-TREATMENT NOT COVERED BY SUBCLASSES C08B, C08C, C08F, C08G or C08H
- C08J2367/00—Characterised by the use of polyesters obtained by reactions forming a carboxylic ester link in the main chain; Derivatives of such polymers
- C08J2367/04—Polyesters derived from hydroxy carboxylic acids, e.g. lactones
-
- C—CHEMISTRY; METALLURGY
- C08—ORGANIC MACROMOLECULAR COMPOUNDS; THEIR PREPARATION OR CHEMICAL WORKING-UP; COMPOSITIONS BASED THEREON
- C08J—WORKING-UP; GENERAL PROCESSES OF COMPOUNDING; AFTER-TREATMENT NOT COVERED BY SUBCLASSES C08B, C08C, C08F, C08G or C08H
- C08J2369/00—Characterised by the use of polycarbonates; Derivatives of polycarbonates
-
- C—CHEMISTRY; METALLURGY
- C08—ORGANIC MACROMOLECULAR COMPOUNDS; THEIR PREPARATION OR CHEMICAL WORKING-UP; COMPOSITIONS BASED THEREON
- C08J—WORKING-UP; GENERAL PROCESSES OF COMPOUNDING; AFTER-TREATMENT NOT COVERED BY SUBCLASSES C08B, C08C, C08F, C08G or C08H
- C08J2405/00—Characterised by the use of polysaccharides or of their derivatives not provided for in groups C08J2401/00 or C08J2403/00
- C08J2405/08—Chitin; Chondroitin sulfate; Hyaluronic acid; Derivatives thereof
-
- C—CHEMISTRY; METALLURGY
- C08—ORGANIC MACROMOLECULAR COMPOUNDS; THEIR PREPARATION OR CHEMICAL WORKING-UP; COMPOSITIONS BASED THEREON
- C08J—WORKING-UP; GENERAL PROCESSES OF COMPOUNDING; AFTER-TREATMENT NOT COVERED BY SUBCLASSES C08B, C08C, C08F, C08G or C08H
- C08J2429/00—Characterised by the use of homopolymers or copolymers of compounds having one or more unsaturated aliphatic radicals, each having only one carbon-to-carbon double bond, and at least one being terminated by an alcohol, ether, aldehydo, ketonic, acetal, or ketal radical; Hydrolysed polymers of esters of unsaturated alcohols with saturated carboxylic acids; Derivatives of such polymer
- C08J2429/02—Homopolymers or copolymers of unsaturated alcohols
- C08J2429/04—Polyvinyl alcohol; Partially hydrolysed homopolymers or copolymers of esters of unsaturated alcohols with saturated carboxylic acids
-
- C—CHEMISTRY; METALLURGY
- C08—ORGANIC MACROMOLECULAR COMPOUNDS; THEIR PREPARATION OR CHEMICAL WORKING-UP; COMPOSITIONS BASED THEREON
- C08J—WORKING-UP; GENERAL PROCESSES OF COMPOUNDING; AFTER-TREATMENT NOT COVERED BY SUBCLASSES C08B, C08C, C08F, C08G or C08H
- C08J2467/00—Characterised by the use of polyesters obtained by reactions forming a carboxylic ester link in the main chain; Derivatives of such polymers
- C08J2467/04—Polyesters derived from hydroxy carboxylic acids, e.g. lactones
-
- C—CHEMISTRY; METALLURGY
- C08—ORGANIC MACROMOLECULAR COMPOUNDS; THEIR PREPARATION OR CHEMICAL WORKING-UP; COMPOSITIONS BASED THEREON
- C08J—WORKING-UP; GENERAL PROCESSES OF COMPOUNDING; AFTER-TREATMENT NOT COVERED BY SUBCLASSES C08B, C08C, C08F, C08G or C08H
- C08J2469/00—Characterised by the use of polycarbonates; Derivatives of polycarbonates
Landscapes
- Health & Medical Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Medicinal Chemistry (AREA)
- General Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Veterinary Medicine (AREA)
- Dermatology (AREA)
- Oral & Maxillofacial Surgery (AREA)
- Transplantation (AREA)
- Epidemiology (AREA)
- Public Health (AREA)
- Animal Behavior & Ethology (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Polymers & Plastics (AREA)
- Organic Chemistry (AREA)
- Dispersion Chemistry (AREA)
- Medicinal Preparation (AREA)
Abstract
The invention provides a cross-linked gel material containing polymer microspheres, a preparation method thereof and an injection filler, wherein the method comprises the following steps: 1) Dissolving aliphatic polyester substances in an organic solvent to serve as an oil phase, wherein the concentration of the aliphatic polyester substances is 5-40wt%; dissolving a surfactant and a biocompatible polymer in water to form a water phase; the molecular weight of the biocompatible macromolecule is 20-400 ten thousand; 2) Mixing the oil phase and the water phase, and then continuously emulsifying to obtain an oil-in-water emulsion; 3) Simultaneously curing and crosslinking the emulsion and a crosslinking agent under an alkaline condition to obtain a crosslinked gel block containing polymer microspheres; 4) And (3) sequentially dialyzing, removing the solvent, sterilizing and the like the crosslinked gel block containing the polymer microspheres to obtain the crosslinked gel material containing the polymer microspheres. The invention simplifies the production process, can improve the production efficiency and save the input cost, and can obtain filler products with smaller cross-linking agent residue.
Description
Technical Field
The invention belongs to the technical field of medical cosmetology, and particularly relates to a crosslinked gel material containing polymer microspheres, a preparation method thereof and an injection filler.
Background
The skin injection usage amount is reported to be the most in the total treatment course number of Chinese medicine and American in 2020, and accounts for more than 60%. The skin injection filling products mainly comprise hyaluronic acid, collagen and polyesters. The dermal filler is an injection type three types of medical devices, and the action principle of the dermal filler is that a material with good biocompatibility is injected into skin tissues to play a role in filling the skin tissues or stimulating tissues to synthesize collagen, so that wrinkles of the skin or contour defects are smoothed.
Dermal filler development history includes: since 1981, the first generation of dermal fillers were animal-derived collagen fillers (e.g., zyderm, cosmoplast, and Sunmex, etc.), which were post-operativeThe effect lasts for a short time (2-4 months), animal-derived risks exist, skin allergy testing needs to be performed one month before the operation, and the use is inconvenient. Beginning in 2003, the second generation dermal fillers were cross-linked hyaluronic acid fillers (e.g., restylane) TM 、
And
etc.) which degrade in the human body to free hyaluronic acid. Hyaluronic acid is originally present in the skin, so that side effects such as skin allergy and the like are remarkably reduced, and the clinical use amount is the largest at present. However, the retention time of the crosslinked hyaluronic acid filler in the body is still relatively short (6-12 months), and the injection must be repeated every 6-12 months, so that frequent operation brings high injection infection risk. Since 2009, the third generation of fillers were artificially synthesized degradable polymeric fillers (e.g., synthetic polymeric fillers)
) Contains polylactic acid (PLA) or Polycaprolactone (PCL) microparticles and is relatively slowly decomposed in the human body (1-4 years). Compared with the first two generations of fillers, the polyester filler belongs to a semi-permanent filler, but has poor biocompatibility and more side effects compared with a cross-linked hyaluronic acid filler.
In order to make up for the defects of the third generation products, a plurality of technical schemes are disclosed at present, wherein polyester microspheres are mainly combined with cross-linked hyaluronic acid, and the polyester microspheres are mixed or wrapped in cross-linked hyaluronic acid gel, so that the semi-permanent characteristics are given to the products, the products have good biocompatibility, and the side effects caused by subcutaneous injection are reduced. For example, patent documents having publication numbers KR1020160052318, WO2017196051A1, CN110559489A and CN111184909A, etc., have a main technical idea of mixing PCL microspheres with crosslinked hyaluronic acid gel by mechanically stirring them.
Specifically, the production process of the above-disclosed technical scheme is emphasized by two steps. The preparation method comprises the following steps of 1, preparing polyester microspheres by an emulsification method (a process flow diagram is shown in figure 1), dissolving polycaprolactone into dichloromethane to serve as an oil phase (marked as dissolution B), dissolving a polyvinyl alcohol surfactant into water to serve as a water phase (marked as dissolution A), injecting the oil phase into the water phase, strongly stirring or homogenizing and emulsifying to form oil-in-water emulsion, and carrying out the procedures of heat preservation and solidification of the microspheres (volatilization of an organic solvent), washing (elution of the surfactant), sieving, irradiation sterilization and the like to obtain the polycaprolactone microspheres.
And 2, preparing the cross-linked hyaluronic acid gel (a process flow chart is shown in figure 2), dissolving hyaluronic acid powder and a NaOH solution in water, adding the NaOH solution containing BDDE, and carrying out cross-linking (heat preservation), PBS dialysis (elution of residual cross-linking agent), sieving (granulation), moist heat sterilization and other processes to obtain cross-linked hyaluronic acid gel particles. Hyaluronic Acid (HA), also known as Hyaluronic acid, is a disaccharide glycosaminoglycan composed of D-glucuronic acid and N-acetylglucosamine; HA is widely distributed in each part of a human body and HAs good hydrophilicity, viscoelasticity, lubricity and biocompatibility; usually, hyaluronic acid is present in the form of a salt, so hyaluronic acid may be referred to as sodium hyaluronate. Crosslinked hyaluronic acid gel: the cross-linked hyaluronic acid is formed by introducing a cross-linking agent to cross-link on the basis of long-chain molecules of hyaluronic acid to form a network structure, and finally exists in the form of a hydrophilic elastomer. BDDE:1, 4-Butanediol diglycidyl ether (1, 4-Butanediol diglycidyl ether, abbreviated as BDDE) is a commonly used hyaluronic acid crosslinking agent.
The technical scheme disclosed above requires at least two steps, namely, two production lines of the polyester microspheres and the crosslinked hyaluronic acid, to obtain the filled product containing the combination of the polyester microspheres and the crosslinked hyaluronic acid gel. Wherein the polyester microspheres are subjected to a drying process, and a plurality of repeated processes such as heat preservation (curing/crosslinking), washing (dialysis), sieving and sterilization processes exist. Therefore, the prior technical scheme has the disadvantages of complex process, low production efficiency and high input cost. In addition, the crosslinked hyaluronic acid dialysis process has limitations in the number of times of dialysis or dialysis time, and is not favorable for further removal of the crosslinking agent residues.
Disclosure of Invention
In view of the above, the present invention provides a crosslinked gel material containing polymer microspheres, a preparation method thereof, and an injection filler, and aims to simplify the production process, improve the production efficiency, save the input cost, and obtain a filler product with less crosslinker residue.
The invention provides a preparation method of a crosslinked gel material containing polymer microspheres, which comprises the following steps:
1) Dissolving aliphatic polyester substances in an organic solvent to serve as an oil phase, wherein the concentration of the aliphatic polyester substances is 5-40wt%; dissolving a surfactant and a biocompatible polymer in water to form a water phase; the molecular weight of the biocompatible macromolecule is 20-400 ten thousand;
2) Mixing the oil phase and the water phase, and then continuously emulsifying to obtain an oil-in-water emulsion;
3) Simultaneously curing and crosslinking the emulsion and a crosslinking agent under an alkaline condition to obtain a crosslinked gel block containing polymer microspheres;
4) And (3) sequentially carrying out dialysis, solvent removal, sieving, moist heat sterilization or irradiation sterilization on the crosslinked gel block containing the polymer microspheres to obtain the crosslinked gel material containing the polymer microspheres.
The preparation method provided by the invention has the advantages of simple process, low cost and high efficiency, and can obtain filler products with less cross-linking agent residue.
Referring to fig. 3, fig. 3 is a schematic process flow diagram of the preparation of a crosslinked hyaluronic acid gel containing polyester microspheres according to some embodiments of the present invention.
In the present example, the aqueous phase was prepared in the dissolution step a, and the oil phase was prepared in the dissolution step B. Wherein, a proper amount of aliphatic polyester and/or modified substances thereof can be taken and added into the organic solvent, and the mixture is preferably magnetically stirred and dissolved at room temperature and 200 +/-50 rpm to obtain an oily solution with the concentration of 5-40wt%, namely the oil phase.
In addition, a proper amount of surfactant is weighed, and can be added into water under the stirring of the rotating speed of 300 +/-100 rpm, the temperature is raised to 95 +/-5 ℃, the mixture is dissolved for 2 +/-0.5 h, and then the temperature is lowered to 18-28 ℃ for later use; preferably, a proper amount of 200-400 ten thousand of hyaluronic acid powder with large molecular weight is weighed, added in batches under stirring, and dissolved for 30-60min to form a solution as a water phase.
In the embodiment of the present invention, the aliphatic polyester-based material is a material for forming polyester microspheres, and is selected from at least one of polylactide, polyglycolide, polycaprolactone, polytrimethylene carbonate, and polydioxanone, or a copolymer obtained by copolymerizing two or more of the monomers of the foregoing polymers, or a modified polymer thereof; specifically Mw =0.5-30 ten thousand. Wherein the polylactide is also named polylactic acid and comprises poly-L-lactide and poly-D, L-lactide.
Polycaprolactone (PCL) is a safe degradable material without toxic and side effects, and can be degraded in vivo to obtain CO 2 And H 2 O; the PCL has good biocompatibility and longer degradation period, and is an ideal filling material for medical and aesthetic plastic surgery. Polytrimethylene carbonate (PTMC) is rubbery at body temperature, has a certain elasticity, is an amorphous polymer, and has no fixed melting point; can be widely used for degradable ligature devices, drug controlled release materials, in-vivo implanted materials, in-vivo supporting materials and the like.
The modified polymer includes but is not limited to polyhydroxyalkanoate, which is one or more of polyhydroxybutyrate, hydroxybutanoic acid valeric acid copolyester, hydroxybutanoic acid caproate copolyester, and poly 3-hydroxybutyrate/4-hydroxybutyrate copolymer.
In the embodiment of the present invention, the organic solvent is selected from one or more of substituted or unsubstituted hydrocarbons, ketones, tetrahydrofuran (THF), ethyl acetate and ethyl lactate, the hydrocarbons include aliphatic hydrocarbons and aromatic hydrocarbons, and the substituted hydrocarbons are preferably halogen substituted compounds; the organic solvent is preferably one or more of dichloromethane, trichloromethane, toluene, acetone, N-methyl pyrrolidone (NMP), methyl ethyl ketone and ethyl acetate, and dichloromethane is the preferred choice. And, the concentration of the aliphatic polyester-based material in the formulated oil phase is 5 to 40wt%, preferably 10 to 30wt%.
In the embodiment of the invention, the surfactant and the biocompatible polymer are dissolved in water, namely the water phase, generally the water for injection is adopted, and the water solution has larger proportion in the technical scheme. The surfactant has certain hydrophilicity, is beneficial to processes such as dissolution and subsequent emulsification, and is preferably polyvinyl alcohol and/or tween, and preferably polyvinyl alcohol (specifically Mw =2.5-15 ten thousand). Preferably, the concentration of polyvinyl alcohol in the aqueous phase is 0.1% to 2.0%, preferably 0.5% by weight.
In the embodiment of the present invention, the biocompatible polymer is one or more of hyaluronic acid, collagen, chitosan, carboxymethyl cellulose derivative, polyamino acid, dextran, and starch, and preferably hyaluronic acid. The molecular weight Mw of the biocompatible macromolecule is preferably 40-300 ten thousand; for example, the molecular weight of the macromolecular hyaluronic acid is 200 to 400 ten thousand, preferably 260 to 300 ten thousand, and a macromolecule such as 20 to 40 ten thousand hyaluronic acid may be used. According to the embodiment of the invention, a pore-foaming agent is optionally added into a polymer raw material to obtain porous microspheres; the amount of the pore-forming agent such as inorganic salts such as calcium chloride can be about 1%.
After the preparation of the water phase and the oil phase is completed, the preferred embodiment of the invention carries out the emulsification process: heating to 20-40 deg.C, stirring or homogenizing at 600-2500pm speed, injecting the oil phase into the water phase with a syringe pump at 30-60ml/min, and emulsifying for 15-30min to obtain oil-in-water (O/W) emulsion. In the embodiment of the invention, the solution viscosity is reduced by heating, and the emulsification effect is improved; and a certain speed is adopted for pumping to ensure the stable transportation of the water-oil ratio.
In the embodiment of the present invention, the stirring or homogenizing emulsification mode may also be other modes, such as ultrasonic treatment. The speed of rotation of the stirrer may be 600 to 2500rpm, preferably 1200 to 1600rpm.
In the preferred embodiment of the invention, a reaction kettle is arranged at 35-90 ℃ and the rotating speed of 200-400rpm, a proper amount of sodium hydroxide is added in batches under stirring, the emulsified solution is adjusted to be alkaline, the cross-linking agent is slowly added in batches, the protective atmosphere is started to purge the liquid level, and the stirring is continuously carried out at the rotating speed of 200 +/-50 rpm, so that the viscoelastic block gel containing the polyester microspheres is obtained.
The temperature range of the heat preservation is preferably controlled to be 35-90 ℃, further preferably 35-55 ℃, and preferably 45 ℃, the emulsion and the cross-linking agent are cured/cross-linked under an alkaline condition, and in a certain temperature range, the higher the temperature is, the faster the cross-linking reaction rate is, the higher the cross-linking degree is, and the larger the viscoelasticity of the product is. Wherein, the added cross-linking agent can be selected from binary epoxy, mainly butanediol diglycidyl ether (BDDE), and the specific dosage is 10-15 percent of the mass of hyaluronic acid; other crosslinking agents, such as those containing aldehyde, sulfone, or imine groups (e.g., by varying the amount of crosslinking agent, the incubation time, and the pH of the reaction system). The reasons for using higher concentrations of the cross-linking agent in embodiments of the present invention include: the reaction system HAs more water phase, so that the HA concentration is low, the solution viscosity is low, and the PCL microspheres with small particle sizes can be obtained through high-speed emulsification and dispersion; on the other hand, the number of effective crosslinks is decreased, and therefore, the concentration of the crosslinking agent is increased (the amount of the crosslinking agent used is increased), and finally, a gel having an appropriate degree of crosslinking is obtained.
The invention preferably adopts protective atmosphere to purge the liquid level, which can accelerate the volatilization of the organic solvent; in this process, the microspheres are cured, the organic solvent is volatilized, and the crosslinking reaction is simultaneously performed to obtain a crosslinked gel mass containing the polymer microspheres. According to the embodiment of the invention, sodium hydroxide can be added to realize an alkaline system (such as pH value of 9-14), and the reaction of hyaluronic acid and BDDE can be promoted. The reaction time for the heat preservation can be 6-12h, preferably 6-8h; the reaction time is relatively long, the crosslinking degree is high, and the crosslinked hyaluronic acid gel with higher viscoelasticity is obtained. Of course, other alkaline agents may be used to adjust the pH, including but not limited to potassium hydroxide (KOH), sodium carbonate (Na) 2 CO 3 ) Ammonia-ammonium chloride (NH) 3 ·H 2 O-NH 4 Cl) basic buffer solution, etc.
The block-shaped cross-linked gel obtained in the embodiment of the invention is cut into pieces so as to remove excessive surfactant, cross-linking agent and organic solvent by dialysis. Specifically, the crosslinked gel mass containing polymeric microspheres is cut into pieces having a size of about 0.5 to 2cm 3 Transferring the irregular block-packed matter into a dialysis device filled with water for injection, dialyzing for 7-11 times, wherein each dialysis time is 60-120min, and obtaining the cross-linked hyaluronic acid gel containing the polyester microspheresAnd (6) gluing. In the embodiment of the invention, the crosslinked gel block containing the polymer microspheres is washed and dialyzed by using water for injection, and the using amount of the water for injection is 20-30 times of the weight of the gel; the water for injection is used as the dialyzate instead of the phosphoric acid solution, so that the excessive phosphate content and the excessive osmotic pressure in the intermediate freeze-dried powder are avoided.
In the embodiment of the invention, dialysis is preferably carried out for 7-11 times, the dialysis time is 60-120min each time, the residual quantity of the cross-linking agent can be ensured to be less than 0.05ppm (the industry standard is less than or equal to 2 ppm), and the harm (periodic red, swollen, itchy and the like at the injection part) caused by the cross-linking agent is further reduced.
For the purpose of further removing residual dichloromethane organic solvent and the like, some embodiments of the invention transfer the dialyzed crosslinked hyaluronic acid gel block containing the polyester microspheres to a tray for drying, preferably freeze-drying in a freeze-dryer, to obtain a freeze-dried block. Exemplary lyophilization conditions include: pre-freezing at-40 + -5 deg.C for 2-3h, sublimation drying at-5-30 deg.C for 8-12h, and resolution drying at 40 + -2 deg.C for 6-8h. Freeze drying is preferred in the embodiment of the invention, which is helpful for maintaining a loose structure of the gel block after freeze drying and is convenient for crushing; other drying means, such as forced air drying, are also possible. In other embodiments, the solvent is removed by concentration. The drying process and the concentration process can remove the organic solvent, and the concentration operation can adjust the water content in the gel, and takes shorter time (the concentration time is 2-4h in total).
The intermediate product in the prior art is generally gel, the gel contains a large amount of water, a large-size storage container is needed, the storage is inconvenient, the crosslinked hyaluronic acid and the polyester microspheres are unstable in water and have certain hydrolysis, and the prepared intermediate product gel has to enter the next production procedure as soon as possible, so that the regulation and control of a production plan are not facilitated.
According to the embodiment of the invention, the polyester microspheres and the cross-linked hyaluronic acid gel are formed simultaneously through one production line, so that the input cost of equipment and a clean area is saved; and the prepared composition of the intermediate product, namely the polyester microspheres and the cross-linked hyaluronic acid, is freeze-dried powder, is easy to store and is convenient for the arrangement of production plans.
Moreover, the freeze drying and the like ensure that the dialysis times are not limited by the content of the cross-linked hyaluronic acid in the final product; the reason is that the more times of dialysis of the obtained gel block is, the larger the water absorption capacity of the cross-linked hyaluronic acid is, the content of the cross-linked hyaluronic acid is gradually reduced, and the expected marked content (such as 15 mg/ml) of the final product cannot be achieved; and the freeze-dried and other dried products are powder, and a certain amount of phosphoric acid buffer solution, normal saline and the like are added for swelling before filling, so that the accurate labeled amount of the cross-linked hyaluronic acid can be obtained. The dialysis procedure of the invention can be unlimited in times and time, so that a filler product with less residual cross-linking agent can be obtained, and side effects caused by the cross-linking agent are greatly reduced.
After the solvent is removed, the invention can be directly sieved or sieved after being crushed. In a preferred embodiment of the present invention, the lyophilized cake is pulverized in a pulverizer, and the powder sieved through a multi-stage sieve (80 mesh/150 mesh/300 mesh) is collected and then mixed with biocompatible small molecule powder such as small molecule hyaluronic acid. Wherein, the addition of micromolecular hyaluronic acid (not crosslinked) can increase the fluidity of the terminal product and effectively reduce the extrusion force in clinical use.
In the embodiment of the invention, a powder sample obtained by mixing biocompatible micromolecules after sieving is taken and subpackaged in a sterilization bag for irradiation sterilization. Wherein, for example, low molecular weight hyaluronic acid is used, which functions as a lubricant to reduce the resistance at the time of injection, and the low molecular weight hyaluronic acid is not subjected to a crosslinking reaction; the molecular weight of hyaluronic acid used at this time may range from 40 to 80 ten thousand.
In some embodiments of the present invention, the wet heat sterilization tank may be used for sterilization, such as sterilization at 121 ℃ for 15min, to obtain a sterile intermediate product; the sterilization mode is more suitable for biocompatible high molecular raw materials with relatively low molecular weight, such as hyaluronic acid raw materials with molecular weight of 20-200 ten thousand. If a moist heat sterilization method is used, the swelling can be performed before the sterilization process. The invention preferably adopts irradiation sterilization (including electron beam sterilization or gamma ray sterilization), which is beneficial to ensuring the structure of the product and the like. The irradiation dose of the irradiation sterilization is preferably 15-25kGy, and a sterile intermediate product (such as cross-linked hyaluronic acid powder containing polyester microspheres) is obtained and stored at room temperature for later use.
After irradiation sterilization, the molecular chain of the crosslinked HA is partially broken, and the in vivo degradation resistance is reduced. The embodiment of the invention selects proper HA initial molecular weight, and the in vivo degradation period is about 2 months after irradiation; the molecular weight of the polyester microspheres is reduced to a certain extent due to irradiation, so that the expected molecular weight range is reached; the process simultaneously completes the sterilization of the HA cross-linked gel and the polyester microspheres.
The degradation period of the components of the cross-linked hyaluronic acid gel in the prior disclosed technical scheme is too long (more than 3 months), which is not beneficial to the stimulation of the regeneration of collagen in skin tissues by the polyester microspheres in a short period (2 months); the action mechanism of the preferred composition is to smooth wrinkles through the physical filling effect of the gel component in the early stage (2 months), and then the polymer particles or microspheres continuously stimulate skin tissues to cause fibroblast hyperplasia and promote collagen secretion, thereby playing a role in correcting skin wrinkles.
The process of the preferred embodiment of the invention adopts a radiation sterilization mode, which not only ensures the sterility level of the product, but also partially degrades the cross-linked hyaluronic acid through radiation, thereby achieving the expected purpose of degradation cycle of about 2 months in vivo.
In order to obtain the pre-encapsulated gel injection, the sterile intermediate product is swelled, and the related buffer solution comprises phosphate buffer solution, normal saline and the like. The specific operation example is as follows: adding the sterile intermediate product into phosphoric acid buffer solution in batches, swelling for 30-60min, preferably stirring at a rotation speed of 200 +/-50 rpm for 10-20min, and uniformly mixing to obtain a cross-linked gel solution containing the polymer microspheres. The swelling can also be carried out by using normal saline (0.9% NaCl), and the swelling time is not greatly influenced within 30-60 min.
The invention provides a cross-linked gel material containing polymer microspheres obtained by the preparation method as described above, wherein the residue of the cross-linking agent is preferably less than 0.05ppm and far lower than the industry standard. The technology is applied to the products: a cross-linked sodium hyaluronate gel comprising polycaprolactone microspheres; the application fields relate to facial reshaping, vocal cord treatment, urinary incontinence, tumor embolism, bone defect repair and the like.
In the embodiment of the invention, under a hundred-grade purification environment, the swelled crosslinked hyaluronic acid gel containing the polyester microspheres is filled into an aseptic prefilled syringe by a filling machine to obtain the filler of the crosslinked hyaluronic acid gel containing the polyester microspheres. Specifically, the mass ratio of the polyester microspheres is 2% -50%, preferably 20% -30%. The filler product is generally 1 ml/piece, 2 ml/piece and the like; the proportion of the polyester microspheres is based on the content of the single filler (comprising cross-linked hyaluronic acid, PBS buffer and PCL microspheres). The invention provides an injection filler, which is prepared by filling the cross-linked gel material containing the polymer microspheres.
In conclusion, the technical scheme of the invention adopts a one-step method, can prepare materials such as cross-linked hyaluronic acid gel containing polyester microspheres, omits a drying procedure in the process of preparing the polyester microspheres by an emulsification method, and combines a plurality of procedures (curing, washing, sieving, sterilizing and the like) in the prior art; in general, the technical scheme can shorten the production period (2-4 d), remarkably improve the production efficiency and greatly reduce the cost input. In addition, the cross-linked hyaluronic acid gel and other materials containing the polyester microspheres prepared by the invention have smaller cross-linking agent residue, are used as injection fillers, and have better effects such as degradation period and the like.
Drawings
FIG. 1 is a process flow diagram of the preparation of conventional polymeric microspheres;
FIG. 2 is a process flow diagram of the preparation of a conventional crosslinked hyaluronic acid gel;
fig. 3 is a schematic process flow diagram of the preparation of a crosslinked hyaluronic acid gel containing polyester microspheres according to some embodiments of the present invention.
Detailed Description
The technical solutions in the embodiments of the present application are clearly and completely described below, and it is obvious that the described embodiments are only a part of the embodiments of the present application, and not all embodiments. All other embodiments, which can be derived by a person skilled in the art from the embodiments given herein without making any creative effort, shall fall within the protection scope of the present application.
In order to better understand the technical content of the invention, specific examples are provided below to further illustrate the invention.
Example 1
S1.1, weighing 8g of polyvinyl alcohol with Mw =3 ten thousand, adding 1000ml of water for injection while stirring at the rotation speed of 300rpm, heating to 95 ℃, stirring for dissolving for 2 hours, and cooling to 20 ℃ for later use; 3g of hyaluronic acid powder having a weight-average molecular weight of 260 ten thousand was weighed, and added in portions with stirring and dissolved to form a solution as an aqueous phase.
S1.2, 40g of Mw =4 ten thousand polycaprolactone is weighed and added into 200ml of dichloromethane, 1% of calcium chloride is mixed, and the mixture is magnetically stirred to be dissolved at room temperature and 200rpm to serve as an oil phase.
S2 emulsification
Heating the water phase to 30 ℃, mechanically stirring at 1300rpm, injecting the oil phase into the water phase by using an injection pump at 40ml/min, and continuously emulsifying for 20min after the injection is finished to form an emulsion of oil-in-water droplets;
s3 Heat preservation (curing/crosslinking)
Setting a reaction kettle at a rotating speed of 300rpm at 45 ℃, adding 8g of sodium hydroxide in batches to enable a reaction system to be in an alkaline state, slowly injecting 0.5g of BDDE in batches, continuously stirring at 200rpm, and starting a nitrogen purging liquid level; and (3) carrying out heat preservation and crosslinking for 8h at the temperature of 45 ℃ to obtain the viscoelastic gel containing the polycaprolactone microspheres.
S4 dialysis (washing)
Cutting the block gel in S3 to a size of about 0.5-2cm 3 The irregular mass of (2) was transferred to a dialysis apparatus containing water for injection, and dialysis was performed 7 times for 120min each time, and the gel mass after dialysis was collected.
S5 drying
And (4) transferring the crosslinked hyaluronic acid gel block containing the polyester microspheres in the S4 into a blast oven to volatilize excessive water, so as to obtain a dried block.
S6 crushing
The dried cake in S5 was put into a pulverizer to be pulverized, and the powder sieved through a multi-stage sieve (sequentially 80 mesh/150 mesh/300 mesh) was collected, followed by mixing 1g of hyaluronic acid having a molecular weight of 60 ten thousand.
S7 radiation sterilization
And (5) subpackaging the samples in the step (S6) into sterilization bags, performing irradiation sterilization, wherein the irradiation dose is 25kGy, obtaining a sterile intermediate product, and storing for later use.
S8 swelling
Weighing a proper amount of the intermediate product in the S7, adding the intermediate product into 156ml of phosphoric acid buffer solution in batches, swelling for 30min, stirring for 15min, and uniformly mixing to obtain the cross-linked hyaluronic acid gel solution containing the polyester microspheres.
S9 filling
And (3) under a hundred-grade purification environment, filling the crosslinked hyaluronic acid gel containing the polyester microspheres in the S8 into an aseptic pre-filling and sealing syringe through a filling machine to obtain the filling agent of the crosslinked hyaluronic acid gel containing the polyester microspheres, wherein the filling agent contains 20% of polyester microspheres, the concentration of the crosslinked hyaluronic acid is 15mg/ml, and the concentration of the non-crosslinked hyaluronic acid is 5mg/ml. The specification of the filling agent is 1 ml/branch or 2 ml/branch, and the proportion of each component is fixed.
Example 2
S1 preparation of water phase and oil phase
S1.1, weighing 8g of polyvinyl alcohol with Mw =3 ten thousand, adding 1000ml of water for injection while stirring at the rotation speed of 300rpm, heating to 95 ℃, stirring for dissolving for 2 hours, and cooling to 20 ℃ for later use; weighing 3.5g of hyaluronic acid powder with the molecular weight of 260 ten thousand, adding the hyaluronic acid powder in batches under stirring, and dissolving the hyaluronic acid powder for 45min to form a solution serving as a water phase;
s1.2 weighing 20g Mw =2 ten thousand polycaprolactone and 20g Mw =8 ten thousand polytrimethylene carbonate, adding the materials into 200ml dichloromethane, magnetically stirring the materials at the room temperature and the rotating speed of 200rpm until the materials are dissolved to obtain an oil phase.
S2 emulsification
Heating the water phase to 30 ℃, mechanically stirring at 1300rpm, injecting the oil phase into the water phase by using an injection pump at 40ml/min, and continuously emulsifying for 20min after the injection is finished to form an oil-in-water droplet emulsion;
s3 Heat preservation (curing/crosslinking)
Setting a reaction kettle at the rotating speed of 300rpm at 45 ℃, adding 8g of sodium hydroxide in 3-5 batches to ensure that a reaction system is in an alkaline state, slowly injecting 0.5g of BDDE in batches, continuously stirring at 150rpm, and starting a nitrogen purging liquid level; and (3) carrying out heat preservation and crosslinking at 45 ℃ for 8h to obtain the viscoelastic gel containing polycaprolactone and polytrimethylene carbonate microspheres.
S4 dialysis (washing)
Cutting the block gel in S3 to about size0.5-2cm 3 The irregular mass of (2) was transferred to a dialysis apparatus containing water for injection, and dialysis was performed for 9 times for 120min each time, and the gel mass after dialysis was collected.
S5 Freeze drying
And transferring the crosslinked hyaluronic acid gel block containing polycaprolactone and polytrimethylene carbonate microspheres in the S4 to a tray, putting the tray into a freeze dryer, and carrying out freeze drying (prefreezing at (-40 ℃) for 2h, sublimation drying at-5-30 ℃ for 8-12h, and resolution drying at 40 ℃ for 6-8 h) to obtain a freeze-dried block.
S6 crushing
The lyophilized cake in S5 was put into a pulverizer to be pulverized, and the powder sieved through a multi-stage sieve (sequentially 80 mesh/150 mesh/300 mesh) was collected, followed by mixing 0.5g of hyaluronic acid having a molecular weight of 60 ten thousand.
S7 radiation sterilization
And (5) subpackaging the samples in the step (S6) into sterilization bags, carrying out irradiation sterilization with the irradiation dose of 25kGy, obtaining sterile intermediate products, and storing for later use.
S8 swelling
Weighing a proper amount of the intermediate in the S7, adding the intermediate into 156ml of phosphoric acid buffer solution in batches, swelling for 30min, stirring for 15min, and uniformly mixing to obtain the cross-linked hyaluronic acid gel solution containing polycaprolactone and polytrimethylene carbonate microspheres.
S9 filling
And (3) under a hundred-grade purification environment, filling the crosslinked hyaluronic acid gel containing polycaprolactone and polytrimethylene carbonate microspheres in the S8 into an aseptic pre-filling and sealing syringe by a filling machine to obtain the filler of the crosslinked hyaluronic acid gel containing polycaprolactone and polytrimethylene carbonate microspheres. It contains 20% of polycaprolactone and polytrimethylene carbonate microspheres, the concentration of cross-linked hyaluronic acid is 17.5mg/ml, and the concentration of non-cross-linked hyaluronic acid is 2.5mg/ml.
Example 3
S1 preparation of water phase and oil phase
S1.1, weighing 8g of polyvinyl alcohol with Mw =3 ten thousand, adding 1000ml of water for injection under stirring at 300rpm, heating to 95 ℃, stirring for dissolving for 2h, and cooling to 20 ℃ for later use; weighing 3g of hyaluronic acid powder with the molecular weight of 260 ten thousand, adding the hyaluronic acid powder in batches under stirring, and dissolving the hyaluronic acid powder for 45min to form a solution serving as a water phase;
s1.2 weighing 60g of polycaprolactone with Mw=2 ten thousand, adding the polycaprolactone into 300ml of dichloromethane, and magnetically stirring the polycaprolactone at a rotating speed of 200rpm under a room temperature condition until the polycaprolactone is dissolved to serve as an oil phase.
S2 emulsification
Heating the water phase to 30 ℃, mechanically stirring at 1300rpm, injecting the oil phase into the water phase by using an injection pump at 40ml/min, and continuously emulsifying for 20min after the injection is finished to form an oil-in-water droplet emulsion;
s3 Heat preservation (curing/crosslinking)
Setting a reaction kettle at the rotation speed of 300rpm at 45 ℃, adding 8g of sodium hydroxide in 3-5 batches to enable a reaction system to be in an alkaline state, slowly injecting 0.5g of BDDE in batches, continuously stirring at 150rpm, and starting a nitrogen purging liquid level; and (4) keeping the temperature at 45 ℃ for crosslinking for 8h to obtain the viscoelastic gel containing the polycaprolactone microspheres.
S4 dialysis (washing)
Cutting the block gel in S3 to a size of about 0.5-2cm 3 The irregular mass of (2) was transferred to a dialysis apparatus containing water for injection, and dialysis was performed for 11 times for 120min each time, and the gel mass after dialysis was collected.
S5 Freeze drying
And (3) transferring the crosslinked hyaluronic acid gel block containing the polycaprolactone microspheres in the S4 to a tray, putting the crosslinked hyaluronic acid gel block into a freeze dryer, and carrying out freeze drying (prefreezing at-40 ℃ for 2h, sublimation drying at-5-30 ℃ for 8-12h, and resolution drying at 40 ℃ for 6-8 h) to obtain a freeze-dried block.
S6 crushing
The lyophilized cake in S5 was put into a pulverizer to be pulverized, and the powder sieved through a multi-stage sieve (sequentially 80 mesh/150 mesh/300 mesh) was collected, followed by mixing 1g of hyaluronic acid having a molecular weight of 60 ten thousand.
S7 radiation sterilization
And (5) subpackaging the samples in the step (S6) into sterilization bags, carrying out irradiation sterilization with the irradiation dose of 25kGy, obtaining sterile intermediate products, and storing for later use.
S8 swelling
Weighing a proper amount of the intermediate product in the S7, adding the intermediate product into 136ml of phosphoric acid buffer solution in batches, swelling for 30min, stirring for 15min, and uniformly mixing to obtain the cross-linked hyaluronic acid gel solution containing the polycaprolactone microspheres.
S9 filling
And (3) under a hundred-grade purification environment, filling the crosslinked hyaluronic acid gel containing the polycaprolactone microspheres in the S8 into an aseptic pre-filled and sealed syringe through a filling machine to obtain the filler of the crosslinked hyaluronic acid gel containing the polycaprolactone microspheres, wherein the filler contains 30% of polyester microspheres, the concentration of the crosslinked hyaluronic acid is 15mg/ml, and the concentration of the non-crosslinked hyaluronic acid is 5mg/ml.
Example 4
Preparation of S1 Water phase and oil phase
S1.1, weighing 8g of polyvinyl alcohol with Mw =3 ten thousand, adding 1000ml of water for injection while stirring at the rotation speed of 300rpm, heating to 95 ℃, stirring for dissolving for 2 hours, cooling to 20 ℃ for later use, weighing 3.5g of hyaluronic acid powder with the molecular weight of 260 ten thousand, and adding and dissolving the hyaluronic acid powder in batches while stirring to form a solution serving as a water phase;
s1.2 weighing 40g of poly-L-lactic acid with Mw =10 ten thousand, adding the poly-L-lactic acid into 200ml of dichloromethane, and magnetically stirring the mixture at the room temperature and the rotation speed of 200rpm until the mixture is dissolved to form an oil phase.
S2 emulsification
Heating the water phase to 30 ℃, mechanically stirring at 1100rpm, injecting the oil phase into the water phase by using an injection pump at 40ml/min, and continuously emulsifying for 20min after the injection is finished to form liquid drops;
s3 Heat preservation (curing/crosslinking)
Setting a reaction kettle at the rotation speed of 300rpm at 45 ℃, adding 8g of sodium hydroxide in 3-5 batches to enable a reaction system to be in an alkaline state, slowly injecting 0.5g of BDDE in batches, continuously stirring at 150rpm, and starting a nitrogen purging liquid level; and (3) carrying out heat preservation and crosslinking for 8h at the temperature of 45 ℃ to obtain the viscoelastic gel containing the poly-L-lactic acid microspheres.
S4 dialysis (washing)
Cutting the block-shaped gel in S3 into pieces with size of about 0.5-2cm 3 The irregular mass of (2) was transferred to a dialysis apparatus containing water for injection, and dialysis was performed for 5 times for 60min each time, and the gel mass after dialysis was collected.
S5 Freeze drying
Transferring the crosslinked hyaluronic acid gel block containing poly-L-lactic acid in S4 to a tray, placing into a freeze dryer, and freeze drying (prefreezing at-40 deg.C for 2h, -5-30 deg.C for sublimation drying for 8-12h, and resolution drying at 40 deg.C for 6-8 h) to obtain lyophilized block.
S6 crushing
The lyophilized cake in S5 was pulverized in a pulverizer, and the powder was sieved through a multi-stage sieve (sequentially 80 mesh/150 mesh/300 mesh), and 40-200 μm particles were collected, followed by mixing 0.5g of hyaluronic acid having a molecular weight of 60 ten thousand.
S7 swelling
Weighing a proper amount of intermediate products in the S6, adding the intermediate products into 156ml of phosphoric acid buffer solution in batches, swelling for 30min, stirring for 15min, and uniformly mixing to obtain the cross-linked hyaluronic acid gel solution containing the poly-L-lactic acid microspheres.
S8 moist Heat Sterilization
And (4) sub-packaging the sample in the step (S7) in a damp-heat sterilization tank, and carrying out damp-heat sterilization at the sterilization temperature of 121 ℃ for 15min to obtain a sterile intermediate product.
S9 filling
And (3) under a hundred-grade purification environment, filling the crosslinked hyaluronic acid gel containing the poly-L-lactic acid microspheres in the S8 into an aseptic pre-filled and sealed syringe by a filling machine to obtain the filling agent of the crosslinked hyaluronic acid gel containing the poly-L-lactic acid microspheres, wherein the filling agent contains 20% of poly-L-lactic acid microspheres, the concentration of the crosslinked hyaluronic acid is 17.5mg/ml, and the concentration of the non-crosslinked hyaluronic acid is 2.5mg/ml.
Example 5
S1 preparation of water phase and oil phase
S1.1, weighing 8g of polyvinyl alcohol with Mw =3 ten thousand, adding 1000ml of water for injection while stirring at the rotation speed of 300rpm, heating to 95 ℃, stirring for dissolving for 2 hours, cooling to 20 ℃ for later use, weighing 3.5g of hyaluronic acid powder with the molecular weight of 40 ten thousand, and adding and dissolving the hyaluronic acid powder in batches while stirring to form a solution serving as a water phase;
s1.2 weighing 40g of poly-L-lactic acid with Mw =15 ten thousand, adding the poly-L-lactic acid into 200ml of dichloromethane, and magnetically stirring the mixture at the room temperature and the rotation speed of 200rpm until the mixture is dissolved to form an oil phase.
S2 emulsification
Heating the water phase to 30 ℃, mechanically stirring at 1200rpm, injecting the oil phase into the water phase by using an injection pump at 40ml/min, and continuously emulsifying for 20min after injection to form liquid drops;
s3 Heat preservation (curing/crosslinking)
Setting a reaction kettle at the rotating speed of 300rpm at 45 ℃, adding 8g of sodium hydroxide in 3-5 batches to enable a reaction system to be in an alkaline state, slowly injecting 0.4g of BDDE in batches, continuously stirring at 150rpm, and starting a nitrogen purging liquid level; and (3) carrying out heat preservation and crosslinking for 12h at the temperature of 45 ℃ to obtain the viscoelastic gel containing the poly-L-lactic acid microspheres.
S4 dialysis (washing)
Cutting the block gel in S3 to a size of about 0.5-2cm 3 The irregular block is transferred to a dialysis device filled with buffer solution, dialysis is carried out for 5 times, each time for 120min, and the gel block after dialysis is collected.
S5 concentration
The S4 gel cake was transferred to a rotary evaporator and concentrated while removing the organic solvent to a final volume of about 200ml. The sample is gel after concentration, and does not need to be added with water for swelling.
S6 sieving
And (4) sieving the concentrated blocks in the S5 by a multi-stage sieve (sequentially passing through 80 meshes/150 meshes/300 meshes), and collecting particles with the particle size of 40-200 mu m.
S7 moist Heat Sterilization
And (4) sub-packaging the sample in the step (S6) in a damp-heat sterilization tank, and carrying out damp-heat sterilization at the sterilization temperature of 121 ℃ for 15min to obtain a sterile intermediate product.
S8 filling
And (3) under a hundred-grade purification environment, filling the crosslinked hyaluronic acid gel containing the poly-L-lactic acid microspheres in the S7 into an aseptic prefilled syringe by using a filling machine to obtain the filler of the crosslinked hyaluronic acid gel containing the poly-L-lactic acid microspheres, wherein the content of the poly-L-lactic acid microspheres is 20%, and the concentration of the crosslinked hyaluronic acid is 17.5mg/ml.
Example 6
Corresponding samples are prepared according to the methods of examples 1 to 4, and are respectively marked as 1#, 2#, 3#, and 4#; the residual amount of BDDE was measured according to the procedure defined in appendix F in accordance with "crosslinked sodium hyaluronate gel for orthopedic surgery" YY/T0961-2014. The BDDE residual test results are shown in table 1.
TABLE 1 BDDE residual test results
As can be seen from Table 1, the dialysis time of the sample No. 1-3 is 2 times that of the sample No. 4, the dialysis frequency is increased by more than 2 times, the residue of the cross-linking agent in the former is one order of magnitude lower than that in the latter, the BDDE residue of the cross-linking agent can be obviously reduced by increasing the dialysis frequency and prolonging the dialysis time, and the biological safety of the product is further improved; and the residual amount of the cross-linking agent gradually decreases with the increase of the dialysis times.
The pushing force was tested according to YY/T0962-2014 Cross-Linked sodium hyaluronate gel for Plastic surgery; the results of the squeeze force test are shown in Table 2.
TABLE 2 Pushing force measurement data
The experimental data in table 2 show that the increase of the content of the small molecular hyaluronic acid is beneficial to improving the fluidity of the product and obviously reducing the extrusion force of the product; on the other hand, when the content of the PCL microspheres is increased, the extrusion force is increased.
As can be seen from the above examples, the process of the present invention comprises: 1) Aliphatic polyester substances are dissolved in an organic solvent to be used as an oil phase, and the concentration of the aliphatic polyester substances is 5-40wt%; dissolving a surfactant and a biocompatible polymer in water to form a water phase; the molecular weight of the biocompatible macromolecule is 20-400 ten thousand; 2) Mixing the oil phase and the water phase, and then continuously emulsifying to obtain an oil-in-water emulsion; 3) Simultaneously curing and crosslinking the emulsion and a crosslinking agent under an alkaline condition to obtain a crosslinked gel block containing polymer microspheres; 4) And (3) dialyzing, removing the solvent, sterilizing and the like the crosslinked gel block containing the polymer microspheres to obtain the crosslinked gel material containing the polymer microspheres. The invention simplifies the production process, can improve the production efficiency and save the input cost, and can obtain filler products with smaller cross-linking agent residue.
The principles and embodiments of the present invention have been described herein using specific examples, which are presented only to assist in understanding the method and its core concepts of the present invention. The foregoing is only a preferred embodiment of the present invention, and it should be noted that there are objectively infinite specific structures due to the limited character expressions, and it will be apparent to those skilled in the art that a plurality of modifications, decorations or changes may be made without departing from the principle of the present invention, and the technical features described above may be combined in a suitable manner; such modifications, variations, or combinations, or other applications of the inventive concepts and solutions as may be employed without such modifications, are intended to be included within the scope of the present invention.
Claims (10)
1. A preparation method of a cross-linked gel material containing polymer microspheres is characterized by comprising the following steps:
1) Dissolving aliphatic polyester substances in an organic solvent to serve as an oil phase, wherein the concentration of the aliphatic polyester substances is 5-40wt%; dissolving a surfactant and a biocompatible polymer in water to form a water phase; the molecular weight of the biocompatible macromolecule is 20-400 ten thousand;
2) Mixing the oil phase and the water phase, and then continuously emulsifying to obtain an oil-in-water emulsion;
3) Simultaneously curing and crosslinking the emulsion and a crosslinking agent under an alkaline condition to obtain a crosslinked gel block containing polymer microspheres;
4) And (3) sequentially carrying out dialysis, solvent removal, sieving, moist heat sterilization or irradiation sterilization on the crosslinked gel block containing the polymer microspheres to obtain the crosslinked gel material containing the polymer microspheres.
2. The method according to claim 1, wherein the aliphatic polyester-based material is at least one selected from the group consisting of polylactide, polyglycolide, polycaprolactone, polytrimethylene carbonate and polydioxanone, or a copolymer obtained by copolymerizing two or more monomers of the above-mentioned polymers, or a modified polymer thereof; the organic solvent is selected from one or more of substituted or unsubstituted hydrocarbons, ketones, tetrahydrofuran, ethyl acetate and ethyl lactate, preferably one or more of dichloromethane, trichloromethane, toluene, acetone, N-methylpyrrolidone, methyl ethyl ketone and ethyl acetate; the concentration of the aliphatic polyester-based substance is preferably 10 to 30% by weight.
3. The method of claim 1, wherein the surfactant is polyvinyl alcohol and/or tween, preferably polyvinyl alcohol; the biocompatible polymer is one or more of hyaluronic acid, collagen, chitosan, carboxymethyl cellulose derivative, polyamino acid, dextran and starch, preferably hyaluronic acid; the molecular weight of the biocompatible polymer is preferably 40 to 300 ten thousand.
4. The method according to any one of claims 1 to 3, wherein the step 2) is specifically: heating to 20-40 deg.C, stirring or homogenizing at 600-2500rpm, adding the oil phase into the water phase, and emulsifying to obtain oil-in-water emulsion.
5. The method according to claim 4, wherein the amount of the cross-linking agent used in step 3) is 10-15% of the mass of the biocompatible polymer, and the curing and cross-linking are performed simultaneously at a temperature of 35-90 ℃ under a protective atmosphere purge and stirring.
6. The method of claim 4, wherein in the step 4), the crosslinked gel mass containing the polymeric microspheres is dialyzed by washing with water for injection, preferably 7-11 times of dialysis, each time for 60-120min.
7. The method according to any one of claims 1 to 3, wherein in step 4), the biocompatible small molecule is mixed after sieving to obtain a powder sample; preferably, the sterile intermediate product is obtained by adopting irradiation sterilization and irradiation dose of 15-25 kGy.
8. The method for preparing the anti-radiation agent according to claim 7, wherein the step 4) further comprises the following steps after the radiation sterilization: and adding the sterile intermediate products into the buffer solution in batches, and swelling for 30-60min to obtain a cross-linked gel solution containing the polymer microspheres.
9. The crosslinked gel material comprising polymeric microspheres obtained by the method according to any one of claims 1 to 8, preferably having a crosslinker residue of less than 0.05ppm.
10. An injection filling agent prepared by filling the cross-linked gel material containing the polymer microspheres of claim 9.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202211280983.XA CN115584037A (en) | 2022-10-19 | 2022-10-19 | Crosslinked gel material containing polymer microspheres, preparation method thereof and injection filler |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202211280983.XA CN115584037A (en) | 2022-10-19 | 2022-10-19 | Crosslinked gel material containing polymer microspheres, preparation method thereof and injection filler |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN115584037A true CN115584037A (en) | 2023-01-10 |
Family
ID=84779840
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN202211280983.XA Pending CN115584037A (en) | 2022-10-19 | 2022-10-19 | Crosslinked gel material containing polymer microspheres, preparation method thereof and injection filler |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN115584037A (en) |
Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN116173294A (en) * | 2023-02-13 | 2023-05-30 | 深圳高性能医疗器械国家研究院有限公司 | Microsphere for injection filling and preparation method thereof |
| CN117298355A (en) * | 2023-11-28 | 2023-12-29 | 常州百瑞吉生物医药股份有限公司 | Coated hyaluronic acid gel composition and preparation method and application thereof |
| CN118557793A (en) * | 2024-05-31 | 2024-08-30 | 南京汇创星美生物科技有限公司 | Facial filler containing hyaluronic acid-PHA (polyhydroxyalkanoate) -crosslinked freeze-dried microspheres and preparation method thereof |
Citations (9)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| KR20170123099A (en) * | 2016-04-28 | 2017-11-07 | 주식회사 한국비엔씨 | Dermal Filler Composition Containing Polycaprolactone And Hyaluronic Acid |
| CN109621003A (en) * | 2018-12-07 | 2019-04-16 | 长春杭盖生物科技有限公司 | A kind of preparation method of the injectable hyaluronic acid sodium gel containing microballoon |
| CN109824919A (en) * | 2019-03-21 | 2019-05-31 | 深港产学研基地 | The preparation method of Sodium Hyaluronate composite high-molecular gel |
| CN112999426A (en) * | 2021-03-03 | 2021-06-22 | 江苏西宏生物医药有限公司 | Chitosan/polycaprolactone composite microsphere gel |
| CN113230451A (en) * | 2021-04-02 | 2021-08-10 | 长春圣博玛生物材料有限公司 | Injectable dermal filler and preparation method thereof |
| CN113616604A (en) * | 2021-10-12 | 2021-11-09 | 北京蓝晶微生物科技有限公司 | Injectable polyhydroxyalkanoate microspheres and preparation method thereof |
| CN113842500A (en) * | 2021-09-24 | 2021-12-28 | 华熙生物科技股份有限公司 | Preparation method and application of degradable polymer microspheres |
| CN113861458A (en) * | 2021-09-24 | 2021-12-31 | 华熙生物科技股份有限公司 | Preparation method of hyaluronic acid filler |
| CN114904049A (en) * | 2022-05-31 | 2022-08-16 | 山东柏佳薇生物科技有限公司 | Polycaprolactone microsphere gel containing hyaluronic acid and collagen and preparation method thereof |
-
2022
- 2022-10-19 CN CN202211280983.XA patent/CN115584037A/en active Pending
Patent Citations (9)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| KR20170123099A (en) * | 2016-04-28 | 2017-11-07 | 주식회사 한국비엔씨 | Dermal Filler Composition Containing Polycaprolactone And Hyaluronic Acid |
| CN109621003A (en) * | 2018-12-07 | 2019-04-16 | 长春杭盖生物科技有限公司 | A kind of preparation method of the injectable hyaluronic acid sodium gel containing microballoon |
| CN109824919A (en) * | 2019-03-21 | 2019-05-31 | 深港产学研基地 | The preparation method of Sodium Hyaluronate composite high-molecular gel |
| CN112999426A (en) * | 2021-03-03 | 2021-06-22 | 江苏西宏生物医药有限公司 | Chitosan/polycaprolactone composite microsphere gel |
| CN113230451A (en) * | 2021-04-02 | 2021-08-10 | 长春圣博玛生物材料有限公司 | Injectable dermal filler and preparation method thereof |
| CN113842500A (en) * | 2021-09-24 | 2021-12-28 | 华熙生物科技股份有限公司 | Preparation method and application of degradable polymer microspheres |
| CN113861458A (en) * | 2021-09-24 | 2021-12-31 | 华熙生物科技股份有限公司 | Preparation method of hyaluronic acid filler |
| CN113616604A (en) * | 2021-10-12 | 2021-11-09 | 北京蓝晶微生物科技有限公司 | Injectable polyhydroxyalkanoate microspheres and preparation method thereof |
| CN114904049A (en) * | 2022-05-31 | 2022-08-16 | 山东柏佳薇生物科技有限公司 | Polycaprolactone microsphere gel containing hyaluronic acid and collagen and preparation method thereof |
Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN116173294A (en) * | 2023-02-13 | 2023-05-30 | 深圳高性能医疗器械国家研究院有限公司 | Microsphere for injection filling and preparation method thereof |
| CN117298355A (en) * | 2023-11-28 | 2023-12-29 | 常州百瑞吉生物医药股份有限公司 | Coated hyaluronic acid gel composition and preparation method and application thereof |
| CN117298355B (en) * | 2023-11-28 | 2024-03-08 | 常州百瑞吉生物医药股份有限公司 | Coated hyaluronic acid gel composition and preparation method and application thereof |
| CN118557793A (en) * | 2024-05-31 | 2024-08-30 | 南京汇创星美生物科技有限公司 | Facial filler containing hyaluronic acid-PHA (polyhydroxyalkanoate) -crosslinked freeze-dried microspheres and preparation method thereof |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN115584037A (en) | Crosslinked gel material containing polymer microspheres, preparation method thereof and injection filler | |
| CN113730652B (en) | Mixed gel for injection and preparation method and application thereof | |
| CN102548590B (en) | Viscoelastic gels as novel fillers | |
| CA2452412C (en) | Biodegradable injectable implants and related methods of manufacture and use | |
| US9345809B2 (en) | Carboxymethylcellulose polyethylene glycol compositions for medical uses | |
| CN104001209B (en) | Microparticles comprising PCL and uses thereof | |
| WO2017136935A1 (en) | Dermal filler composed of macroporous chitosan microbeads and cross-linked hyaluronic acid | |
| EP0402031A2 (en) | Injectable polymeric bodies | |
| WO1997004012A1 (en) | Polysaccharide gel composition | |
| CN113058074B (en) | Temperature-sensitive filler composition | |
| CN108653817A (en) | A kind of preparation and application of novel collagen stimulant | |
| CN107397980B (en) | Anti-adhesion composition for coating tissue repair film and using method thereof | |
| TWI840478B (en) | Crosslinked polysaccharides and related methods | |
| CN110755686A (en) | Injectable inducible bone repair material and preparation method and application thereof | |
| WO2020185074A1 (en) | Particulate gel for the treatment of a bleeding in the sinus or nasal cavity | |
| CN114395164B (en) | Polysaccharide composite gel and preparation method and application thereof | |
| CN115282339B (en) | Crosslinked hyaluronic acid/hydroxyapatite injectable material, preparation method and application | |
| CN115671388A (en) | Performance-adjustable silk protein injectable microsphere gel and preparation method thereof | |
| KR102070878B1 (en) | Method of preparing muk-gel for filler procedure by cross-linking hyaluronic acid using electron-beam without cross-linking agent | |
| CN115536872A (en) | Crosslinked gel material containing polymer microspheres and preparation method thereof | |
| CN112996551B (en) | Irradiated agarose, compositions thereof, and related methods | |
| CN115887760A (en) | Preparation process of L-polylactic acid for injection | |
| TW202432088A (en) | Crosslinked polysaccharides and related methods | |
| CN116271224A (en) | Soft tissue regeneration filler and preparation method thereof | |
| CN118320184A (en) | Injectable exosome-loaded silk fibroin microsphere-carboxymethyl cellulose composite gel 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 |