CN117815175A - Bile salt-lecithin-musk ketone nano micelle and preparation method and application thereof - Google Patents
Bile salt-lecithin-musk ketone nano micelle and preparation method and application thereof Download PDFInfo
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- CN117815175A CN117815175A CN202311548307.0A CN202311548307A CN117815175A CN 117815175 A CN117815175 A CN 117815175A CN 202311548307 A CN202311548307 A CN 202311548307A CN 117815175 A CN117815175 A CN 117815175A
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- China
- Prior art keywords
- lecithin
- bile salt
- nano
- musk ketone
- micelle
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Links
- 239000000693 micelle Substances 0.000 title claims abstract description 80
- 229940067137 musk ketone Drugs 0.000 title claims abstract description 79
- 210000000941 bile Anatomy 0.000 title claims abstract description 31
- 238000002360 preparation method Methods 0.000 title claims abstract description 25
- 239000003833 bile salt Substances 0.000 claims abstract description 95
- FACFHHMQICTXFZ-UHFFFAOYSA-N 2-(2-phenylimidazo[1,2-a]pyridin-3-yl)ethanamine Chemical compound N1=C2C=CC=CN2C(CCN)=C1C1=CC=CC=C1 FACFHHMQICTXFZ-UHFFFAOYSA-N 0.000 claims abstract description 60
- IIZPXYDJLKNOIY-JXPKJXOSSA-N 1-palmitoyl-2-arachidonoyl-sn-glycero-3-phosphocholine Chemical compound CCCCCCCCCCCCCCCC(=O)OC[C@H](COP([O-])(=O)OCC[N+](C)(C)C)OC(=O)CCC\C=C/C\C=C/C\C=C/C\C=C/CCCCC IIZPXYDJLKNOIY-JXPKJXOSSA-N 0.000 claims abstract description 34
- 229940067606 lecithin Drugs 0.000 claims abstract description 34
- 235000010445 lecithin Nutrition 0.000 claims abstract description 34
- 239000000787 lecithin Substances 0.000 claims abstract description 34
- 238000000034 method Methods 0.000 claims abstract description 21
- 239000003814 drug Substances 0.000 claims abstract description 19
- 239000002994 raw material Substances 0.000 claims abstract description 3
- WTJKGGKOPKCXLL-RRHRGVEJSA-N phosphatidylcholine Chemical compound CCCCCCCCCCCCCCCC(=O)OC[C@H](COP([O-])(=O)OCC[N+](C)(C)C)OC(=O)CCCCCCCC=CCCCCCCCC WTJKGGKOPKCXLL-RRHRGVEJSA-N 0.000 claims description 19
- OKKJLVBELUTLKV-UHFFFAOYSA-N Methanol Chemical group OC OKKJLVBELUTLKV-UHFFFAOYSA-N 0.000 claims description 18
- JQWAHKMIYCERGA-UHFFFAOYSA-N (2-nonanoyloxy-3-octadeca-9,12-dienoyloxypropoxy)-[2-(trimethylazaniumyl)ethyl]phosphinate Chemical compound CCCCCCCCC(=O)OC(COP([O-])(=O)CC[N+](C)(C)C)COC(=O)CCCCCCCC=CCC=CCCCCC JQWAHKMIYCERGA-UHFFFAOYSA-N 0.000 claims description 14
- 239000007853 buffer solution Substances 0.000 claims description 12
- ILXAOQAXSHVHTM-UHFFFAOYSA-M sodium;2-amino-2-(hydroxymethyl)propane-1,3-diol;chloride Chemical compound [Na+].[Cl-].OCC(N)(CO)CO ILXAOQAXSHVHTM-UHFFFAOYSA-M 0.000 claims description 10
- ALHUZKCOMYUFRB-UHFFFAOYSA-N muskone Natural products CC1CCCCCCCCCCCCC(=O)C1 ALHUZKCOMYUFRB-UHFFFAOYSA-N 0.000 claims description 9
- JAJWGJBVLPIOOH-IZYKLYLVSA-M sodium taurocholate Chemical group [Na+].C([C@H]1C[C@H]2O)[C@H](O)CC[C@]1(C)[C@@H]1[C@@H]2[C@@H]2CC[C@H]([C@@H](CCC(=O)NCCS([O-])(=O)=O)C)[C@@]2(C)[C@@H](O)C1 JAJWGJBVLPIOOH-IZYKLYLVSA-M 0.000 claims description 9
- 239000002904 solvent Substances 0.000 claims description 9
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 claims description 8
- ALHUZKCOMYUFRB-OAHLLOKOSA-N Muscone Chemical compound C[C@@H]1CCCCCCCCCCCCC(=O)C1 ALHUZKCOMYUFRB-OAHLLOKOSA-N 0.000 claims description 8
- 238000002156 mixing Methods 0.000 claims description 7
- WEVYAHXRMPXWCK-UHFFFAOYSA-N Acetonitrile Chemical compound CC#N WEVYAHXRMPXWCK-UHFFFAOYSA-N 0.000 claims description 6
- DHMQDGOQFOQNFH-UHFFFAOYSA-N Glycine Chemical compound NCC(O)=O DHMQDGOQFOQNFH-UHFFFAOYSA-N 0.000 claims description 4
- 239000007924 injection Substances 0.000 claims description 4
- 238000002347 injection Methods 0.000 claims description 4
- XOAAWQZATWQOTB-UHFFFAOYSA-N taurine Chemical group NCCS(O)(=O)=O XOAAWQZATWQOTB-UHFFFAOYSA-N 0.000 claims description 4
- 239000006185 dispersion Substances 0.000 claims description 3
- 238000001035 drying Methods 0.000 claims description 3
- 229940045946 sodium taurodeoxycholate Drugs 0.000 claims description 3
- YXHRQQJFKOHLAP-FVCKGWAHSA-M sodium;2-[[(4r)-4-[(3r,5r,8r,9s,10s,12s,13r,14s,17r)-3,12-dihydroxy-10,13-dimethyl-2,3,4,5,6,7,8,9,11,12,14,15,16,17-tetradecahydro-1h-cyclopenta[a]phenanthren-17-yl]pentanoyl]amino]ethanesulfonate Chemical compound [Na+].C([C@H]1CC2)[C@H](O)CC[C@]1(C)[C@@H]1[C@@H]2[C@@H]2CC[C@H]([C@@H](CCC(=O)NCCS([O-])(=O)=O)C)[C@@]2(C)[C@@H](O)C1 YXHRQQJFKOHLAP-FVCKGWAHSA-M 0.000 claims description 3
- WBWWGRHZICKQGZ-HZAMXZRMSA-N taurocholic acid Chemical compound C([C@H]1C[C@H]2O)[C@H](O)CC[C@]1(C)[C@@H]1[C@@H]2[C@@H]2CC[C@H]([C@@H](CCC(=O)NCCS(O)(=O)=O)C)[C@@]2(C)[C@@H](O)C1 WBWWGRHZICKQGZ-HZAMXZRMSA-N 0.000 claims description 3
- JLPULHDHAOZNQI-ZTIMHPMXSA-N 1-hexadecanoyl-2-(9Z,12Z-octadecadienoyl)-sn-glycero-3-phosphocholine Chemical compound CCCCCCCCCCCCCCCC(=O)OC[C@H](COP([O-])(=O)OCC[N+](C)(C)C)OC(=O)CCCCCCC\C=C/C\C=C/CCCCC JLPULHDHAOZNQI-ZTIMHPMXSA-N 0.000 claims description 2
- 229940124599 anti-inflammatory drug Drugs 0.000 claims description 2
- 238000004090 dissolution Methods 0.000 claims description 2
- 239000012467 final product Substances 0.000 claims description 2
- 229940040145 liniment Drugs 0.000 claims description 2
- 239000000865 liniment Substances 0.000 claims description 2
- 239000006210 lotion Substances 0.000 claims description 2
- 239000002504 physiological saline solution Substances 0.000 claims description 2
- 229940083466 soybean lecithin Drugs 0.000 claims description 2
- 229960003080 taurine Drugs 0.000 claims description 2
- 239000004471 Glycine Substances 0.000 claims 1
- 239000002260 anti-inflammatory agent Substances 0.000 claims 1
- 239000002245 particle Substances 0.000 abstract description 38
- 229940079593 drug Drugs 0.000 abstract description 12
- 238000005538 encapsulation Methods 0.000 abstract description 9
- 238000009826 distribution Methods 0.000 abstract description 8
- 239000002552 dosage form Substances 0.000 abstract description 5
- 239000000825 pharmaceutical preparation Substances 0.000 abstract description 2
- XEKOWRVHYACXOJ-UHFFFAOYSA-N Ethyl acetate Chemical compound CCOC(C)=O XEKOWRVHYACXOJ-UHFFFAOYSA-N 0.000 description 27
- 230000001965 increasing effect Effects 0.000 description 24
- 238000012360 testing method Methods 0.000 description 14
- 239000000243 solution Substances 0.000 description 10
- 239000013558 reference substance Substances 0.000 description 7
- 239000000203 mixture Substances 0.000 description 6
- 239000000523 sample Substances 0.000 description 6
- 238000005063 solubilization Methods 0.000 description 6
- 230000007928 solubilization Effects 0.000 description 6
- 238000005303 weighing Methods 0.000 description 6
- 229940093761 bile salts Drugs 0.000 description 5
- 238000005119 centrifugation Methods 0.000 description 5
- 239000012488 sample solution Substances 0.000 description 5
- NRHMKIHPTBHXPF-TUJRSCDTSA-M sodium cholate Chemical compound [Na+].C([C@H]1C[C@H]2O)[C@H](O)CC[C@]1(C)[C@@H]1[C@@H]2[C@@H]2CC[C@H]([C@@H](CCC([O-])=O)C)[C@@]2(C)[C@@H](O)C1 NRHMKIHPTBHXPF-TUJRSCDTSA-M 0.000 description 5
- 238000009210 therapy by ultrasound Methods 0.000 description 5
- 238000000861 blow drying Methods 0.000 description 4
- 230000000694 effects Effects 0.000 description 4
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 4
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 description 3
- 238000004458 analytical method Methods 0.000 description 3
- 238000001514 detection method Methods 0.000 description 3
- 238000010586 diagram Methods 0.000 description 3
- 238000005189 flocculation Methods 0.000 description 3
- 230000016615 flocculation Effects 0.000 description 3
- 230000036571 hydration Effects 0.000 description 3
- 238000006703 hydration reaction Methods 0.000 description 3
- 238000011835 investigation Methods 0.000 description 3
- 238000004445 quantitative analysis Methods 0.000 description 3
- 238000003860 storage Methods 0.000 description 3
- RUDATBOHQWOJDD-UHFFFAOYSA-N (3beta,5beta,7alpha)-3,7-Dihydroxycholan-24-oic acid Natural products OC1CC2CC(O)CCC2(C)C2C1C1CCC(C(CCC(O)=O)C)C1(C)CC2 RUDATBOHQWOJDD-UHFFFAOYSA-N 0.000 description 2
- 241000402754 Erythranthe moschata Species 0.000 description 2
- 239000012491 analyte Substances 0.000 description 2
- 239000000872 buffer Substances 0.000 description 2
- 239000012159 carrier gas Substances 0.000 description 2
- 230000008859 change Effects 0.000 description 2
- 230000003247 decreasing effect Effects 0.000 description 2
- 238000002296 dynamic light scattering Methods 0.000 description 2
- 238000002290 gas chromatography-mass spectrometry Methods 0.000 description 2
- 238000003384 imaging method Methods 0.000 description 2
- 238000012417 linear regression Methods 0.000 description 2
- 238000005259 measurement Methods 0.000 description 2
- 239000002105 nanoparticle Substances 0.000 description 2
- RZJRJXONCZWCBN-UHFFFAOYSA-N octadecane Chemical compound CCCCCCCCCCCCCCCCCC RZJRJXONCZWCBN-UHFFFAOYSA-N 0.000 description 2
- 238000005457 optimization Methods 0.000 description 2
- 239000000047 product Substances 0.000 description 2
- 238000011002 quantification Methods 0.000 description 2
- 150000003839 salts Chemical class 0.000 description 2
- 239000012047 saturated solution Substances 0.000 description 2
- 239000011550 stock solution Substances 0.000 description 2
- 239000004094 surface-active agent Substances 0.000 description 2
- HSINOMROUCMIEA-FGVHQWLLSA-N (2s,4r)-4-[(3r,5s,6r,7r,8s,9s,10s,13r,14s,17r)-6-ethyl-3,7-dihydroxy-10,13-dimethyl-2,3,4,5,6,7,8,9,11,12,14,15,16,17-tetradecahydro-1h-cyclopenta[a]phenanthren-17-yl]-2-methylpentanoic acid Chemical class C([C@@]12C)C[C@@H](O)C[C@H]1[C@@H](CC)[C@@H](O)[C@@H]1[C@@H]2CC[C@]2(C)[C@@H]([C@H](C)C[C@H](C)C(O)=O)CC[C@H]21 HSINOMROUCMIEA-FGVHQWLLSA-N 0.000 description 1
- BHQCQFFYRZLCQQ-UHFFFAOYSA-N (3alpha,5alpha,7alpha,12alpha)-3,7,12-trihydroxy-cholan-24-oic acid Natural products OC1CC2CC(O)CCC2(C)C2C1C1CCC(C(CCC(O)=O)C)C1(C)C(O)C2 BHQCQFFYRZLCQQ-UHFFFAOYSA-N 0.000 description 1
- OHERYIXJQJIISS-UHFFFAOYSA-N 3-Methylpentadecan-2-one Chemical group CCCCCCCCCCCCC(C)C(C)=O OHERYIXJQJIISS-UHFFFAOYSA-N 0.000 description 1
- OKTJSMMVPCPJKN-UHFFFAOYSA-N Carbon Chemical compound [C] OKTJSMMVPCPJKN-UHFFFAOYSA-N 0.000 description 1
- 239000004380 Cholic acid Substances 0.000 description 1
- RYGMFSIKBFXOCR-UHFFFAOYSA-N Copper Chemical compound [Cu] RYGMFSIKBFXOCR-UHFFFAOYSA-N 0.000 description 1
- DGABKXLVXPYZII-UHFFFAOYSA-N Hyodeoxycholic acid Natural products C1C(O)C2CC(O)CCC2(C)C2C1C1CCC(C(CCC(O)=O)C)C1(C)CC2 DGABKXLVXPYZII-UHFFFAOYSA-N 0.000 description 1
- 206010061218 Inflammation Diseases 0.000 description 1
- 206010028980 Neoplasm Diseases 0.000 description 1
- BHTRKEVKTKCXOH-UHFFFAOYSA-N Taurochenodesoxycholsaeure Natural products OC1CC2CC(O)CCC2(C)C2C1C1CCC(C(CCC(=O)NCCS(O)(=O)=O)C)C1(C)CC2 BHTRKEVKTKCXOH-UHFFFAOYSA-N 0.000 description 1
- 239000007983 Tris buffer Substances 0.000 description 1
- 230000001133 acceleration Effects 0.000 description 1
- 239000004480 active ingredient Substances 0.000 description 1
- 238000004220 aggregation Methods 0.000 description 1
- 230000002776 aggregation Effects 0.000 description 1
- 230000004075 alteration Effects 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 239000003613 bile acid Substances 0.000 description 1
- 230000004071 biological effect Effects 0.000 description 1
- 230000005540 biological transmission Effects 0.000 description 1
- 229910052799 carbon Inorganic materials 0.000 description 1
- 210000000748 cardiovascular system Anatomy 0.000 description 1
- 239000000969 carrier Substances 0.000 description 1
- 210000003169 central nervous system Anatomy 0.000 description 1
- 238000012512 characterization method Methods 0.000 description 1
- 238000006243 chemical reaction Methods 0.000 description 1
- 229960001091 chenodeoxycholic acid Drugs 0.000 description 1
- RUDATBOHQWOJDD-BSWAIDMHSA-N chenodeoxycholic acid Chemical compound C([C@H]1C[C@H]2O)[C@H](O)CC[C@]1(C)[C@@H]1[C@@H]2[C@@H]2CC[C@H]([C@@H](CCC(O)=O)C)[C@@]2(C)CC1 RUDATBOHQWOJDD-BSWAIDMHSA-N 0.000 description 1
- BHQCQFFYRZLCQQ-OELDTZBJSA-N cholic acid Chemical compound C([C@H]1C[C@H]2O)[C@H](O)CC[C@]1(C)[C@@H]1[C@@H]2[C@@H]2CC[C@H]([C@@H](CCC(O)=O)C)[C@@]2(C)[C@@H](O)C1 BHQCQFFYRZLCQQ-OELDTZBJSA-N 0.000 description 1
- 229960002471 cholic acid Drugs 0.000 description 1
- 235000019416 cholic acid Nutrition 0.000 description 1
- 238000005352 clarification Methods 0.000 description 1
- 150000001875 compounds Chemical class 0.000 description 1
- 238000010276 construction Methods 0.000 description 1
- 229910052802 copper Inorganic materials 0.000 description 1
- 239000010949 copper Substances 0.000 description 1
- KXGVEGMKQFWNSR-UHFFFAOYSA-N deoxycholic acid Natural products C1CC2CC(O)CCC2(C)C2C1C1CCC(C(CCC(O)=O)C)C1(C)C(O)C2 KXGVEGMKQFWNSR-UHFFFAOYSA-N 0.000 description 1
- 238000011161 development Methods 0.000 description 1
- 238000007865 diluting Methods 0.000 description 1
- 238000010790 dilution Methods 0.000 description 1
- 239000012895 dilution Substances 0.000 description 1
- 238000010494 dissociation reaction Methods 0.000 description 1
- 230000005593 dissociations Effects 0.000 description 1
- 239000000839 emulsion Substances 0.000 description 1
- 230000002708 enhancing effect Effects 0.000 description 1
- 238000002474 experimental method Methods 0.000 description 1
- 238000009472 formulation Methods 0.000 description 1
- 239000007789 gas Substances 0.000 description 1
- 238000004817 gas chromatography Methods 0.000 description 1
- 238000010438 heat treatment Methods 0.000 description 1
- DGABKXLVXPYZII-SIBKNCMHSA-N hyodeoxycholic acid Chemical compound C([C@H]1[C@@H](O)C2)[C@H](O)CC[C@]1(C)[C@@H]1[C@@H]2[C@@H]2CC[C@H]([C@@H](CCC(O)=O)C)[C@@]2(C)CC1 DGABKXLVXPYZII-SIBKNCMHSA-N 0.000 description 1
- 230000004054 inflammatory process Effects 0.000 description 1
- 150000002500 ions Chemical class 0.000 description 1
- 238000004519 manufacturing process Methods 0.000 description 1
- 238000004949 mass spectrometry Methods 0.000 description 1
- 239000000463 material Substances 0.000 description 1
- 238000012986 modification Methods 0.000 description 1
- 230000004048 modification Effects 0.000 description 1
- 239000002539 nanocarrier Substances 0.000 description 1
- 239000013642 negative control Substances 0.000 description 1
- 229940038384 octadecane Drugs 0.000 description 1
- 150000002894 organic compounds Chemical class 0.000 description 1
- 238000005191 phase separation Methods 0.000 description 1
- XAEFZNCEHLXOMS-UHFFFAOYSA-M potassium benzoate Chemical compound [K+].[O-]C(=O)C1=CC=CC=C1 XAEFZNCEHLXOMS-UHFFFAOYSA-M 0.000 description 1
- 239000002244 precipitate Substances 0.000 description 1
- 230000008569 process Effects 0.000 description 1
- 230000002035 prolonged effect Effects 0.000 description 1
- 238000010926 purge Methods 0.000 description 1
- 239000012088 reference solution Substances 0.000 description 1
- 230000001105 regulatory effect Effects 0.000 description 1
- 238000011160 research Methods 0.000 description 1
- 238000005070 sampling Methods 0.000 description 1
- 230000035945 sensitivity Effects 0.000 description 1
- 239000011780 sodium chloride Substances 0.000 description 1
- AAYACJGHNRIFCT-YRJJIGPTSA-M sodium glycochenodeoxycholate Chemical compound [Na+].C([C@H]1C[C@H]2O)[C@H](O)CC[C@]1(C)[C@@H]1[C@@H]2[C@@H]2CC[C@H]([C@@H](CCC(=O)NCC([O-])=O)C)[C@@]2(C)CC1 AAYACJGHNRIFCT-YRJJIGPTSA-M 0.000 description 1
- OABYVIYXWMZFFJ-ZUHYDKSRSA-M sodium glycocholate Chemical compound [Na+].C([C@H]1C[C@H]2O)[C@H](O)CC[C@]1(C)[C@@H]1[C@@H]2[C@@H]2CC[C@H]([C@@H](CCC(=O)NCC([O-])=O)C)[C@@]2(C)[C@@H](O)C1 OABYVIYXWMZFFJ-ZUHYDKSRSA-M 0.000 description 1
- 159000000000 sodium salts Chemical class 0.000 description 1
- 230000003381 solubilizing effect Effects 0.000 description 1
- 238000013112 stability test Methods 0.000 description 1
- 238000010025 steaming Methods 0.000 description 1
- 239000000126 substance Substances 0.000 description 1
- 238000006467 substitution reaction Methods 0.000 description 1
- 230000008685 targeting Effects 0.000 description 1
- BHTRKEVKTKCXOH-LBSADWJPSA-N tauroursodeoxycholic acid Chemical compound C([C@H]1C[C@@H]2O)[C@H](O)CC[C@]1(C)[C@@H]1[C@@H]2[C@@H]2CC[C@H]([C@@H](CCC(=O)NCCS(O)(=O)=O)C)[C@@]2(C)CC1 BHTRKEVKTKCXOH-LBSADWJPSA-N 0.000 description 1
- 239000012085 test solution Substances 0.000 description 1
- 230000007704 transition Effects 0.000 description 1
- LENZDBCJOHFCAS-UHFFFAOYSA-N tris Chemical compound OCC(N)(CO)CO LENZDBCJOHFCAS-UHFFFAOYSA-N 0.000 description 1
- 238000002604 ultrasonography Methods 0.000 description 1
- RUDATBOHQWOJDD-UZVSRGJWSA-N ursodeoxycholic acid Chemical compound C([C@H]1C[C@@H]2O)[C@H](O)CC[C@]1(C)[C@@H]1[C@@H]2[C@@H]2CC[C@H]([C@@H](CCC(O)=O)C)[C@@]2(C)CC1 RUDATBOHQWOJDD-UZVSRGJWSA-N 0.000 description 1
- 229960001661 ursodiol Drugs 0.000 description 1
- 238000012795 verification Methods 0.000 description 1
- 230000000007 visual effect Effects 0.000 description 1
Classifications
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K9/00—Medicinal preparations characterised by special physical form
- A61K9/10—Dispersions; Emulsions
- A61K9/107—Emulsions ; Emulsion preconcentrates; Micelles
- A61K9/1075—Microemulsions or submicron emulsions; Preconcentrates or solids thereof; Micelles, e.g. made of phospholipids or block copolymers
-
- 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
- A61K31/12—Ketones
- A61K31/122—Ketones having the oxygen directly attached to a ring, e.g. quinones, vitamin K1, anthralin
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K47/00—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient
- A61K47/06—Organic compounds, e.g. natural or synthetic hydrocarbons, polyolefins, mineral oil, petrolatum or ozokerite
- A61K47/24—Organic compounds, e.g. natural or synthetic hydrocarbons, polyolefins, mineral oil, petrolatum or ozokerite containing atoms other than carbon, hydrogen, oxygen, halogen, nitrogen or sulfur, e.g. cyclomethicone or phospholipids
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K47/00—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient
- A61K47/06—Organic compounds, e.g. natural or synthetic hydrocarbons, polyolefins, mineral oil, petrolatum or ozokerite
- A61K47/28—Steroids, e.g. cholesterol, bile acids or glycyrrhetinic acid
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P25/00—Drugs for disorders of the nervous system
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P29/00—Non-central analgesic, antipyretic or antiinflammatory agents, e.g. antirheumatic agents; Non-steroidal antiinflammatory drugs [NSAID]
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P35/00—Antineoplastic agents
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P9/00—Drugs for disorders of the cardiovascular system
Landscapes
- Health & Medical Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Medicinal Chemistry (AREA)
- General Health & Medical Sciences (AREA)
- Veterinary Medicine (AREA)
- Public Health (AREA)
- Pharmacology & Pharmacy (AREA)
- Animal Behavior & Ethology (AREA)
- General Chemical & Material Sciences (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Epidemiology (AREA)
- Organic Chemistry (AREA)
- Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
- Engineering & Computer Science (AREA)
- Molecular Biology (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Biophysics (AREA)
- Oil, Petroleum & Natural Gas (AREA)
- Rheumatology (AREA)
- Neurology (AREA)
- Neurosurgery (AREA)
- Biomedical Technology (AREA)
- Heart & Thoracic Surgery (AREA)
- Cardiology (AREA)
- Pain & Pain Management (AREA)
- Dispersion Chemistry (AREA)
- Medicinal Preparation (AREA)
Abstract
The invention provides a bile salt-lecithin-musk ketone nano micelle, and a preparation method and application thereof, and belongs to the field of pharmaceutical preparations. The bile salt-lecithin-musk ketone nano micelle is prepared from bile salt, lecithin and musk ketone serving as raw materials; the mole ratio of the bile salt to the lecithin to the musk ketone is 15: (9-11): (12-15). The musk ketone nano micelle prepared by the method is spherical, has no adhesion, uniform particle size distribution, high encapsulation efficiency and drug loading, good stability, can obviously improve the solubility of musk ketone, improves the bioavailability, and provides a good dosage form for clinical application of musk ketone.
Description
Technical Field
The invention belongs to the field of pharmaceutical preparations, and particularly relates to a bile salt-lecithin-musk ketone nano micelle, and a preparation method and application thereof.
Background
Muscone (Muscone) is an organic compound of formula C 16 H 30 O, molecular weight 238.42. Musk ketone is the main active ingredient of rare traditional Chinese medicine Musk (Musk), and the academic name is 3-methylpentadecanone. Musk ketone has various biological activities of protecting central nervous system, regulating cardiovascular system, resisting inflammation, resisting tumor, etc., but is volatile, poor in water solubility and low in bioavailability, so that it is limited in development and utilization as medicine.
In recent years, nano medicine carrying becomes a research hot spot, and has wide application in the aspects of improving medicine solubility and stability, improving bioavailability, enhancing targeting property and the like. However, the preparation of nano-dosage forms generally has the problems of complicated process, poor repeatability, high cost, low drug loading rate, low encapsulation efficiency and the like, and the industrialized production is difficult, and in addition, the safety of nano-carriers is also questioned. Therefore, a simple, cheap, green and safe preparation method of nanometer dosage forms needs to be searched for, and a musk ketone nanometer preparation with excellent performance is prepared so as to improve the application of the musk ketone nanometer preparation.
Disclosure of Invention
The invention aims to provide a bile salt-lecithin-musk ketone nano micelle, and a preparation method and application thereof.
The invention provides a bile salt-lecithin-musk ketone nano micelle, which is prepared from bile salt, lecithin and musk ketone as raw materials; the mole ratio of the bile salt to the lecithin to the musk ketone is 15: (9-11): (12-15).
Further, the mole ratio of the bile salt, the lecithin and the musk ketone is 15:9:15.
further, the bile salt is taurine-type bile salt, glycine-type bile salt or free-type bile salt;
preferably, the bile salt is a taurinate type bile salt.
Further, the taurocholate is sodium taurocholate or sodium taurodeoxycholate.
Further, the lecithin is phosphatidylcholine, soybean lecithin or egg yolk lecithin;
preferably, the lecithin is phosphatidylcholine or egg yolk lecithin.
Further, the nano micelle is prepared by adopting a film dispersion method.
The invention also provides a method for preparing the bile salt-lecithin-musk ketone nano micelle, which comprises the following steps:
(1) Dissolving bile salt, lecithin and muscone with solvent, mixing, and drying;
(2) Re-dissolving and ultrasonic treating to obtain the final product.
Further, the method comprises the steps of,
in the step (1), the solvent is methanol, ethanol or acetonitrile;
and/or in the step (2), the solvent used for re-dissolution is physiological saline, PBS buffer solution or Tris-NaCl buffer solution;
and/or, in the step (2), the ultrasonic time is 10-30 min.
Further, the method comprises the steps of,
in the step (2), the volume mass ratio of the redissolved solvent to the total weight of the bile salt, the lecithin and the musk ketone is (10-100): 1.
The invention also provides the application of the bile salt-lecithin-musk ketone nano micelle in preparing anti-inflammatory drugs;
preferably, the medicament is an injection, a liniment or a lotion.
In the present invention, bile Salt (BS) is a sodium salt or potassium salt of Bile acid compounds such as taurocholate.
The musk ketone nano micelle prepared by the method is spherical, has no adhesion, uniform particle size distribution, high encapsulation efficiency and drug loading, good stability, can obviously improve the solubility of musk ketone, improves the bioavailability, and provides a good dosage form for clinical application of musk ketone.
It should be apparent that, in light of the foregoing, various modifications, substitutions and alterations can be made herein without departing from the spirit and scope of the invention as defined by the appended claims.
The above-described aspects of the present invention will be described in further detail below with reference to specific embodiments in the form of examples. It should not be understood that the scope of the above subject matter of the present invention is limited to the following examples only. All techniques implemented based on the above description of the invention are within the scope of the invention.
Drawings
FIG. 1 shows the particle size distribution of BS-L-M-NMs prepared in example 1.
FIG. 2 shows TEM imaging of BS-L-M-NMs prepared in example 1.
FIG. 3 is a GC-MS chromatogram of muscone.
FIG. 4 shows the effect of different storage times at room temperature on the stability of the bile salt-lecithin-muscone nanomicelle according to the invention.
FIG. 5 shows particle size (A) and polydispersity (B) of various bile salt-lecithin-muscone nano-micelles.
FIG. 6 shows the particle size distribution diagram (A) and the conversion diagram (B) of the nano-micelle formed by musk ketone with different proportions.
FIG. 7 is a morphology diagram of BS-L-M-NMs prepared with different molar ratios of BS, L, M, in order of (A) 15:0:3, (B) 15:5:3, (C) 15:9:3, (D) 15:13:3, (E) 15:18:3, (F) 0:9:3.
FIG. 8 shows the effect of increasing the proportion of lecithin on the particle size and polydispersity of the prepared BS-L-M-NMs, with a fixed bile salt content of 15mM, and a musk ketone content of 0mM (A), 3mM (B), 9mM (C) and 15mM (D) in this order.
Detailed Description
The materials and equipment used in the embodiments of the present invention are all known products and are obtained by purchasing commercially available products.
EXAMPLE 1 preparation of the bile salt-lecithin-musk ketone nanomicelle of the invention
Sodium taurocholate (sodium cholate) is a Bile Salt (BS) and lecithin (lecithin, L) is used as a carrier to prepare musk ketone (M) nano-micelle (nanomicelle, NMs), wherein the lecithin is egg yolk lecithin, the content of Phosphatidylcholine (PC) in the egg yolk lecithin is about 80%, and the relative molecular weight of PC is calculated as 760. Precisely weighing BS, L and M according to the molar ratio of BS to L to M of 15:9:15, dissolving with 10 times of methanol, vortex mixing uniformly, and N 2 And (5) blow-drying. And then redissolving the mixture by using a certain volume of Tris-NaCl buffer solution until the final concentration of bile salt is 15mM, and performing ultrasonic treatment for 10min to obtain the bile salt-lecithin-musk ketone nano micelle (BS-L-M-NMs).
EXAMPLE 2 preparation of bile salt-lecithin-musk ketone nanomicelle according to the invention
Sodium taurodeoxycholate (NaTDC) which is a Bile Salt (BS) and lecithin (lecithin, L) which is a natural surfactant are used as carriers to prepare muskone (M) nano-micelle (nanomicelle, NMs), wherein the lecithin is egg yolk lecithin, the content of Phosphatidylcholine (PC) in the egg yolk lecithin is about 80%, and the relative molecular weight of the PC is calculated as 760. Precisely weighing BS, L and M according to the molar ratio of BS to L to M of 15:9:15, dissolving with 10 times of methanol, vortex mixing uniformly, and N 2 And (5) blow-drying. And then redissolving the mixture by using a certain volume of Tris-NaCl buffer solution until the final concentration of bile salt is 15mM, and performing ultrasonic treatment for 10min to obtain the bile salt-lecithin-musk ketone nano micelle (BS-L-M-NMs).
EXAMPLE 3 preparation of bile salt-lecithin-musk ketone nanomicelle according to the invention
Sodium taurocholate (sodium cholate) is a Bile Salt (BS) and lecithin (lecithin, L) is used as a carrier to prepare musk ketone (M) nano-micelle (nanomicelle, NMs), wherein the lecithin is egg yolk lecithin, the content of Phosphatidylcholine (PC) in the egg yolk lecithin is about 80%, and the relative molecular weight of PC is calculated as 760. Precisely weighing BS, L and M according to the molar ratio of BS to L to M of 15:9:15, dissolving with 10 times of ethanol, vortex mixing uniformly, and N 2 And (5) blow-drying. And then redissolving the mixture by using a certain volume of Tris-NaCl buffer solution until the final concentration of bile salt is 15mM, and performing ultrasonic treatment for 10min to obtain the bile salt-lecithin-musk ketone nano micelle (BS-L-M-NMs).
EXAMPLE 4 preparation of bile salt-lecithin-musk ketone nanomicelle according to the invention
Sodium taurocholate (sodium cholate) is a Bile Salt (BS) and lecithin (lecithin, L) is used as a carrier to prepare musk ketone (M) nano-micelle (nanomicelle, NMs), wherein the lecithin is egg yolk lecithin, the content of Phosphatidylcholine (PC) in the egg yolk lecithin is about 80%, and the relative molecular weight of PC is calculated as 760. Precisely weighing BS, L and M according to the molar ratio of BS to L to M of 15:9:15, dissolving with 10 times of methanol, vortex mixing uniformly, and N 2 And (5) blow-drying. And then redissolving the mixture by using a certain volume of PBS buffer solution until the final concentration of bile salt is 15mM, and performing ultrasonic treatment for 10min to obtain the bile salt-lecithin-musk ketone nano micelle (BS-L-M-NMs).
EXAMPLE 5 preparation of bile salt-lecithin-musk ketone nanomicelle according to the invention
Sodium taurocholate (sodium cholate) is a Bile Salt (BS) and lecithin (lecithin, L) is used as a carrier to prepare musk ketone (M) nano-micelle (nanomicelle, NMs), wherein the lecithin is egg yolk lecithin, the content of Phosphatidylcholine (PC) in the egg yolk lecithin is about 80%, and the relative molecular weight of PC is calculated as 760. Precisely weighing BS, L and M according to the molar ratio of BS to L to M of 15:9:15, dissolving with 10 times of methanol, mixing by vortex, and spin-drying by a spin-steaming instrument. And then redissolving the mixture by using a certain volume of Tris-NaCl buffer solution until the final concentration of bile salt is 15mM, and performing ultrasonic treatment for 10min to obtain the bile salt-lecithin-musk ketone nano micelle (BS-L-M-NMs).
The beneficial effects of the present invention are demonstrated by specific test examples below.
Test example 1 characterization of the bile salt-lecithin-muscone nanomicelle of the invention
1. The average particle size and zeta potential of the bile salt-lecithin-muscone nano-micelle (BS-L-M-NMs) prepared in example 1 (the bile salt is specifically sodium taurocholate) were measured by dynamic light scattering. As a result of the dynamic light scattering measurement, the average particle diameter of BS-L-M-NMs was 11nm, the particle diameter distribution was uniform, PDI was less than 0.1, and the zeta potential of BS-L-M-NMs was-21.6 mV.
2. The sample BS-L-M-NMs prepared in example 1 was diluted 1-fold with PBS buffer solution, gently dropped on a copper mesh coated with an ultra-thin carbon film, air-dried at room temperature, and placed under a Transmission Electron Microscope (TEM) to observe the microscopic morphology of BS-L-M-NMs at 5nm and 20nm, respectively. The JEM-2100FTEM acceleration voltage was 200kV and imaging analysis was performed using GatanMicroscopy Suite Software (Las Vegas, NV, USA). Under the TEM 20nm scale, the particle size distribution of BS-L-M-NMs is uniform (left in figure 2), and under the 5nm visual field, the appearance of BS-L-M-NMs is spherical and has no adhesion (right in figure 2), so that the nano micelle has good formability.
Test example 2, musk ketone quantitative methodology verification results
1. Quantitative analysis gas chromatography method: rxi-5Sil MS column (30.0mX1250 μm,0.25 μm, restek Co., USA); the temperature of the sample inlet is 250 ℃; the programmed temperature is 120 ℃, kept for 2min, and heated to 280 ℃ at a heating rate of 40 ℃/min, and kept for 2min. The carrier gas is He gas, the carrier gas pressure is 120kPa (constant pressure), no split flow exists, the total flow is 20.0mL/min, the column flow is 1.48mL/min, the linear speed is 45.6cm/sec, the purge flow is 6.0mL/min, and the sample injection amount is 1 mu L.
Mass spectrometry method: EI ion source, source temperature is 220 ℃, interface temperature is 270 ℃, detector voltage absolute value is 1kV, scanning threshold is set to 200cps, collection mode is SIM, collection m/z 254.30,5.26-8.00 min is 3.50-5.25 min, and collection m/z 238.20.
2. Taking appropriate amount of muscone reference substance, precisely weighing, dissolving with ethyl acetate, and preparing into 5mg.mL -1 Is a stock solution of (a). Gradually diluting with ethyl acetate to obtain reference substance solution with serial concentration gradient. Musk ketone nano-micelles (BS-L-M-NMs) were prepared as described in example 1. Precisely sucking 10 mu L of the middle layer, adding 990 mu L of ethyl acetate, swirling, centrifuging for 10min with 12000g, and collecting the ethyl acetate layer to obtain a nanometer preparation sample solution. According to the method described in example 1, without using muscone, a blank nano micelle was prepared, 10. Mu.L of the intermediate layer was precisely sucked, 990. Mu.L of ethyl acetate was added, vortexed, 12000g was centrifuged for 10min, and the ethyl acetate layer was taken to prepare a negative control solution. In addition, 1mL of Tris-NaCl buffer solution is taken, 1mg of musk ketone is added, vortex is carried out, ultrasound is carried out for 30min,12000g is carried out for 10min, the middle layer is the saturated solution of musk ketone, 10 mu L of the middle layer is precisely absorbed, 990 mu L of ethyl acetate is added, vortex is carried out, 12000g is carried out for 10min, and the ethyl acetate layer is taken, thus obtaining the test sample of musk ketone saturated solution. The solubilization of musk ketone by nano-micelle was evaluated.
The Internal Standard (IS) compound IS octadecane (C18), which IS prepared by ethyl acetate to a concentration of 5mg/mL of IS stock solution, diluted to 50 mug.mL -1 And (5) standby application. Precisely sucking the reference substance solution and the sample solution, respectively adding equal volumes of IS solution to obtain a series of IS-containing reference substance solution and sample solution with concentration gradients, etc.
3. Investigation of specificity
And (3) respectively taking a reference substance solution containing an internal standard, a test substance solution and a negative reference solution without the internal standard, carrying out GC-MS analysis, and recording a chromatogram.
4. Linearity and sensitivity investigation
And analyzing the reference substance solution containing IS at each concentration, and carrying out linear regression by taking the musk ketone peak area/IS peak area as an ordinate and taking the concentration change as an abscissa to obtain a standard curve and a linear range. The lower limit of detection (LOD) is the concentration of the measured component with a signal to noise ratio (S/N) of about 3, and the lower limit of quantification (LLOQ) is the concentration of the measured component with a S/N of about 10.
5. Precision test
In the linear range of the measured components, taking high, medium and low concentrations for daily precision investigation, continuously injecting each concentration for six times, recording peak areas, and expressing the results by Relative Standard Deviation (RSD).
6. Repeatability test
After musk ketone nano micelle (BS-L-M-NMs) was prepared as described in example 1, 5 parts of test solution of musk ketone nano preparation was prepared in parallel as described in "2", sample injection analysis was performed, peak areas were recorded, and the result was expressed as RSD.
7. Stability test
Taking the same sample solution, preserving at 4 ℃, respectively sampling and measuring at 0, 2, 4, 8 and 12 hours, recording peak areas, and expressing the result as RSD.
8. The musk ketone quantitative method has good specificity, the positions of musk ketone peaks in the reference substance solution and the sample solution are consistent, the components such as BS and L in the sample have no influence on musk ketone content measurement, and the peak shape is good (figure 3). The linear regression equation is y=0.0309x+0.00442, r 2 = 0.9991, the lower limit of detection and the lower limit of quantification are 0.25 μg·ml respectively -1 And 0.5. Mu.g.mL -1 Linear range of 0.5-100 mug.mL -1 The daily precision RSD of high, medium and low concentration is 0.64%, 0.42% and 0.53%, the repeatability RSD is 7.56%, the stability RSD is 1.8%, and the quantitative requirement of musk ketone is met. Good repeatability also indicates that the musk ketone nano micelle preparation process has good stability.
Test example 3 determination of muscone solubility
6 batches of musk ketone nano-micelles were prepared according to the method described in example 1, the analysis was performed by injecting samples according to the quantitative method established in test example 2, the peak area ratio (analyte peak area/internal standard peak area) was substituted into the standard curve, the concentration of the analyte was determined, and the musk ketone content in each batch of nano-preparation was calculated. The concentration of musk ketone in the nano micelle is 2.45mg/mL, and the concentration in Tris-NaCl buffer solution is lower than the detection lower limit (figure 3), which cannot be quantitatively analyzed, so that the nano micelle can greatly improve the solubility of musk ketone.
Test example 4, determination of encapsulation efficiency and drug loading
According to the quantitative result of test example 3, the Encapsulation Efficiency (EE) is 57% and the Drug Loading (DL) is 13% calculated according to the formula (1) and the formula (2), which shows that the encapsulation efficiency and the drug loading of the musk ketone nano micelle prepared by the invention are good.
Note that: w (W) 0 For the total amount of musk ketone added, W 1 The content of musk ketone in the nano particles is W 2 Is the total mass of the nano particles.
Test example 5 stability of BS-L-M-NMs
Multiple batches of musk ketone nano-micelles were prepared and their stability was examined as described in example 1.
Centrifuging for 30min at a low speed of 2000g and a high speed of 15000g, measuring particle size and PDI before and after centrifugation, and observing the centrifugal stability of the nano micelle. After centrifugation, the BS-L-M-NMs is still in a colorless transparent state, flocculation, phase separation, demulsification and other phenomena do not occur, and the average particle size and PDI before and after centrifugation do not have obvious changes, so that the BS-L-M-NMs has good centrifugal stability.
After the preparation is stored for one week at different temperatures of-20 ℃,4 ℃, 25 ℃, 40 ℃ and the like, the appearance property and the average particle size of the BS-L-M-NMs are not obviously changed, PDI is slightly increased, but is still less than 0.2, which indicates that the preparation has good stability at different temperatures.
After storage at room temperature for 0d, 1d, 2d, 3d, 5d and 7d, the appearance of BS-L-M-NMs of the invention was not significantly changed, and the particle size and PDI were slightly increased (FIG. 4), suggesting that there may be slight flocculation of BS-L-M-NMs with prolonged storage time. When the salt concentration in the system is increased, the stability of the nano-micelle can be obviously improved, flocculation is slowed down, and the hydration result by using Tris-NaCl buffer (25mM Tris;500mM NaCl) is better than that by using normal saline (figure 4), so that the Tris-NaCl buffer with higher salt content is selected for hydration in the invention.
The nano micelle formed by the small molecular surfactant is easy to depolymerize after being diluted, and the medicine can be released, so that the influence of different concentrations and different dilution factors on BS-L-M-NMs is examined. The critical micelle concentration of BS is about 10mM, when the volume of the redissolution solvent is adjusted to ensure that BS: L: M=30:18:30 (mM), the particle size is not obviously changed, and is 11.45+/-0.06 nm, PDI is slightly increased, and is 0.208+/-0.005, so that the requirement of nano micelle is met.
The nano-micelle is diluted, the nano-micelle has an increasing trend, which is possibly related to the thickness of a hydration film, and also possibly related to the dissociation of part of BS in BS-L-M-NMs and the aggregation of L-M-NMs, but when the nano-micelle is diluted to be far lower than the critical micelle concentration of BS (BS=0.2 mM), the nano-micelle can still exist stably, and the average particle size is smaller than 30nm. The nano micelle with high, medium and low concentration can exist stably after being stored for one week. The above examination shows that the stability of BS-L-M-NMs is good.
Test example 6 influence of different bile salts on physicochemical Properties of BS-L-M-NMs
In addition to selecting sodium taurocholate (NaTC) as a bile salt to construct nano-micelles, the invention also attempts to construct nano-micelles by using other common bile salts such as sodium cholate (NaC), sodium glycocholate (NaGC), sodium glycochenodeoxycholate (NaGCDC), sodium taurocholate (NaTDC) and the like according to the method of the embodiment 1. The molar ratio of fixed bile salts to lecithin was 15:9 (15 mM:9 mM), the ratio of musk ketone (M) (C1: 0mM, C2:3mM, C3:9mM, C4:15 mM) was gradually increased, and the particle size (size) and Polydispersity (PDI) of the nano-micelle were as shown in FIG. 5. When the proportion of musk ketone is low (C1 and C2), the size difference of micelle formed by different bile salts is not large, as the proportion of musk ketone is increased, the particle size of nano micelle is reduced along with the increase of the molecular size of bile salt, and the particle size is ordered as NaC > NaGC (approximately) NaGCDC > NaTC (approximately) NaTDC (figure 5A); naC, naGC, naGCDC, etc. (FIG. 5B) because the micelle changes from small to large particle size (FIG. 6A) as the musk ketone ratio increases, the present invention speculates that the micelle transitions from cake to sphere (FIG. 6B). In the concentration range of the experiment, the size of the micelle formed by the taurinated bile salt (NaTC and NaTDC) is small, the particle size distribution is uniform, and the micelle is obviously superior to free bile salt and glycin bile salt. In addition, bile acids such as ursodeoxycholic acid, hyodeoxycholic acid, tauroursodeoxycholic acid, chenodeoxycholic acid, cholic acid, etc. which are not salified also participate in the construction of nano-micelles, but white precipitates are formed due to poor water solubility of themselves, and nano-micelles are not formed. The above results demonstrate that using taurine bile salts is suitable for constructing musk ketone nano-micelles.
Test example 7 influence of different prescriptions on physicochemical Properties of BS-L-M-NMs
In addition, the present invention investigated the effect of different prescriptions on the physicochemical properties of BS-L-M-NMs, and prepared BS-L-M-NMs according to the method described in example 1. FIG. 7 is a morphology of BS-L-M-NMs prepared at different molar ratios BS, L, M.
From fig. 7, it can be seen that BS and L act synergistically to increase the solubility of M. When only BS is added (the molar ratio of BS, L and M is 15:0:3 in FIG. 7A), BS and M form emulsion with average particle size (523.5+/-8.501) nm, PDI is 0.178+/-0.009, particle size is large, encapsulation efficiency is poor, demulsification is carried out after centrifugation, and M floats on the upper layer of the BS solution; when L is added only, L itself is poor in water solubility and cannot be uniformly dispersed in water, and layering phenomenon occurs after centrifugation (FIG. 7F, molar ratio of BS, L and M is 0:9:3); only when BS and L are in the proper proportion, a clear and transparent musk ketone nano-preparation with small particle size can be formed (figures 7B-7D). Because the polarity of M is very small, compared with BS, L plays a main solubilization role on M, and in order to improve the drug loading, the proportion of L needs to be increased as much as possible, but the solubility of L per se is poor, and BS is needed to be added for solubilization, so that the invention further examines the influence of different prescription proportions on the nano micelle in order to obtain the nano micelle with high drug loading and good stability.
As shown in fig. 8 and table 1: when no M was added (m=0 mM), BS and L formed mixed nano-micelles, and when 1.8> BS: L >0.8, all samples were clear and transparent in appearance, the average particle size increased stepwise with increasing proportion of L, increasing from 8nm to 30nm, and pdi was less than 0.3 (fig. 8A); when a certain amount of M (M concentration is 3mM during preparation) is added, when the ratio of 3> BS to L is greater than 0.8, the particle size of the nano micelle is obviously increased along with the increase of the proportion of L, the particle size is obviously increased from 8nm to 160nm, the increase is obviously increased compared with the case of not adding M (FIG. 8B), the appearance is gradually changed from clarification to opacification, but the PDI is less than 0.3; further increasing the M ratio (M concentration at 9mM and 15mM at the time of preparation), the average particle size and PDI of the nano-micelle both show a trend of decreasing and increasing with increasing L ratio, but the trend of particle size change transits from U-shaped to V-shaped with increasing M (figures 8C and 8D), indicating that the suitable BS: L ratio range is smaller and smaller. This is consistent with theoretical speculation, L plays a major role in solubilization of M, and when the ratio of M is high and L is insufficient for solubilization, BS plays a major role in solubilization, and BS cannot significantly reduce the surface tension of L, so that the particle size of the formed nano-micelle is large, and at this time, BS-L-M-NMs with small size and BS-M-NMs with large and unstable particle size exist in the preparation, so that PDI is large. From FIGS. 8C-8D, the theoretical molar ratio of L to M in the formulation is about 0.6 to about 0.7 for increased drug loading. In the BS-L system, the larger the proportion of L is, the more advantageous for solubilizing M, but the higher the proportion of L is (BS: L < 1.15), the larger the particle size of the nano-micelle formed by BS-L (FIG. 8A), and as M is added, the value of BS: L needs to be further decreased (FIGS. 8B-8D), because BS plays a role in splitting and dispersing L, and the addition of M may limit the dispersing effect of BS. To obtain a nano-micelle with small particle size, good stability and high drug loading and encapsulation efficiency, BS: l=5:3 (15 mm:9 mm) was finally determined.
To further increase the drug loading and verify the pre-optimization results, BS: l=15 mM:9mM was fixed, the theoretical concentration of M (12-30 mM) was gradually increased, and the particle size and PDI were measured. The results showed that the average particle size of the nano-micelles gradually increased with increasing M, PDI increased significantly, and PDI was greater than 0.3 when M was higher than 18mM at a final concentration, and the appearance became milky gradually (Table 2). And finally optimizing the molar ratio of BS to L to M to be 15:9:15. 3 batches of nano-micelles are prepared in parallel according to the optimized prescription, the particle size and PDI are measured, the appearance is observed, and the optimized result is further verified. The 3 batches of nano-micelles are clear and transparent in appearance, the average particle size (11.35+/-0.66) nm and the PDI (PDI) of 0.07+/-0.03, and the prescription optimization result is proved to be reliable.
TABLE 1 influence of different molar ratios BS, L, M on the Properties of musk ketone nanomicelle BS-L-M-NMs
TABLE 2 influence of different M amounts on the properties of musk ketone nanomicelle BS-L-M-NMs
In conclusion, the musk ketone nano micelle prepared by the method of preparing specific bile salt and lecithin through a film dispersion method is clear and transparent, is spherical, has no adhesion, uniform particle size distribution, high encapsulation efficiency and drug loading rate and good stability, can obviously improve the solubility of musk ketone, improves the bioavailability of musk ketone, and provides a good dosage form for clinical application of musk ketone.
Claims (10)
1. A bile salt-lecithin-muscone nano micelle, which is characterized in that: it is prepared from bile salt, lecithin and musk ketone as raw materials; the mole ratio of the bile salt to the lecithin to the musk ketone is 15: (9-11): (12-15).
2. The bile salt-lecithin-muscone nano-micelle according to claim 1, wherein: the mole ratio of the bile salt to the lecithin to the musk ketone is 15:9:15.
3. the bile salt-lecithin-muscone nano-micelle according to claim 1 or 2, characterized in that: the bile salt is taurine type bile salt, glycine type bile salt or free type bile salt;
preferably, the bile salt is a taurinate type bile salt.
4. A bile salt-lecithin-muscone nano-micelle according to claim 3, characterized in that: the taurocholate is sodium taurocholate or sodium taurodeoxycholate.
5. The bile salt-lecithin-muscone nano-micelle according to claim 1 or 2, characterized in that: the lecithin is phosphatidylcholine, soybean lecithin or egg yolk lecithin;
preferably, the lecithin is phosphatidylcholine or egg yolk lecithin.
6. The bile salt-lecithin-muscone nano-micelle according to claim 1 or 2, characterized in that: the nano micelle is prepared by adopting a film dispersion method.
7. A method for preparing the bile salt-lecithin-muscone nano-micelle according to any one of claims 1 to 6, characterized in that: it comprises the following steps:
(1) Dissolving bile salt, lecithin and muscone with solvent, mixing, and drying;
(2) Re-dissolving and ultrasonic treating to obtain the final product.
8. The method according to claim 7, wherein:
in the step (1), the solvent is methanol, ethanol or acetonitrile;
and/or in the step (2), the solvent used for re-dissolution is physiological saline, PBS buffer solution or Tris-NaCl buffer solution;
and/or, in the step (2), the ultrasonic time is 10-30 min.
9. The method according to claim 8, wherein:
in the step (2), the volume mass ratio of the redissolved solvent to the total weight of the bile salt, the lecithin and the musk ketone is (10-100): 1.
10. Use of a bile salt-lecithin-muscone nano-micelle according to any one of claims 1 to 6 in the preparation of an anti-inflammatory drug;
preferably, the medicament is an injection, a liniment or a lotion.
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