CN117752607A - Oral micelle taking ginseng active ingredient as auxiliary material and preparation method and application thereof - Google Patents
Oral micelle taking ginseng active ingredient as auxiliary material and preparation method and application thereof Download PDFInfo
- Publication number
- CN117752607A CN117752607A CN202311819562.4A CN202311819562A CN117752607A CN 117752607 A CN117752607 A CN 117752607A CN 202311819562 A CN202311819562 A CN 202311819562A CN 117752607 A CN117752607 A CN 117752607A
- Authority
- CN
- China
- Prior art keywords
- ginsenoside
- micelle
- ginseng
- polysaccharide
- oral
- 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
- 239000000693 micelle Substances 0.000 title claims abstract description 140
- 241000208340 Araliaceae Species 0.000 title claims abstract description 87
- 235000005035 Panax pseudoginseng ssp. pseudoginseng Nutrition 0.000 title claims abstract description 87
- 235000003140 Panax quinquefolius Nutrition 0.000 title claims abstract description 87
- 235000008434 ginseng Nutrition 0.000 title claims abstract description 87
- 239000000463 material Substances 0.000 title claims abstract description 35
- 239000004480 active ingredient Substances 0.000 title claims abstract description 19
- 238000002360 preparation method Methods 0.000 title abstract description 25
- 229930182494 ginsenoside Natural products 0.000 claims abstract description 97
- 229940089161 ginsenoside Drugs 0.000 claims abstract description 89
- RCINICONZNJXQF-MZXODVADSA-N taxol Chemical compound O([C@@H]1[C@@]2(C[C@@H](C(C)=C(C2(C)C)[C@H](C([C@]2(C)[C@@H](O)C[C@H]3OC[C@]3([C@H]21)OC(C)=O)=O)OC(=O)C)OC(=O)[C@H](O)[C@@H](NC(=O)C=1C=CC=CC=1)C=1C=CC=CC=1)O)C(=O)C1=CC=CC=C1 RCINICONZNJXQF-MZXODVADSA-N 0.000 claims abstract description 89
- 229930012538 Paclitaxel Natural products 0.000 claims abstract description 85
- 229960001592 paclitaxel Drugs 0.000 claims abstract description 85
- 229920001282 polysaccharide Polymers 0.000 claims abstract description 74
- 239000005017 polysaccharide Substances 0.000 claims abstract description 74
- 150000004676 glycans Chemical class 0.000 claims abstract description 65
- 239000003814 drug Substances 0.000 claims abstract description 47
- ZDZOTLJHXYCWBA-VCVYQWHSSA-N N-debenzoyl-N-(tert-butoxycarbonyl)-10-deacetyltaxol Chemical compound O([C@H]1[C@H]2[C@@](C([C@H](O)C3=C(C)[C@@H](OC(=O)[C@H](O)[C@@H](NC(=O)OC(C)(C)C)C=4C=CC=CC=4)C[C@]1(O)C3(C)C)=O)(C)[C@@H](O)C[C@H]1OC[C@]12OC(=O)C)C(=O)C1=CC=CC=C1 ZDZOTLJHXYCWBA-VCVYQWHSSA-N 0.000 claims abstract description 29
- 229960003668 docetaxel Drugs 0.000 claims abstract description 29
- 229940079593 drug Drugs 0.000 claims abstract description 24
- 230000000259 anti-tumor effect Effects 0.000 claims abstract description 20
- 238000000034 method Methods 0.000 claims abstract description 17
- 238000010521 absorption reaction Methods 0.000 claims abstract description 15
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 claims description 20
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 15
- 239000007788 liquid Substances 0.000 claims description 10
- 102000004169 proteins and genes Human genes 0.000 claims description 10
- 108090000623 proteins and genes Proteins 0.000 claims description 10
- 238000001914 filtration Methods 0.000 claims description 8
- 238000009210 therapy by ultrasound Methods 0.000 claims description 8
- 238000002390 rotary evaporation Methods 0.000 claims description 7
- 108090000526 Papain Proteins 0.000 claims description 6
- 239000004365 Protease Substances 0.000 claims description 6
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 claims description 6
- 102000004139 alpha-Amylases Human genes 0.000 claims description 6
- 108090000637 alpha-Amylases Proteins 0.000 claims description 6
- 229940024171 alpha-amylase Drugs 0.000 claims description 6
- 239000012528 membrane Substances 0.000 claims description 6
- 229940055729 papain Drugs 0.000 claims description 6
- 235000019834 papain Nutrition 0.000 claims description 6
- 238000001035 drying Methods 0.000 claims description 4
- 229920002472 Starch Polymers 0.000 claims description 3
- 229920002678 cellulose Polymers 0.000 claims description 3
- 239000001913 cellulose Substances 0.000 claims description 3
- 238000004440 column chromatography Methods 0.000 claims description 3
- 238000001816 cooling Methods 0.000 claims description 3
- 239000012153 distilled water Substances 0.000 claims description 3
- 238000010828 elution Methods 0.000 claims description 3
- 238000000605 extraction Methods 0.000 claims description 3
- 239000002244 precipitate Substances 0.000 claims description 3
- 230000001376 precipitating effect Effects 0.000 claims description 3
- 239000011780 sodium chloride Substances 0.000 claims description 3
- 239000008107 starch Substances 0.000 claims description 3
- 235000019698 starch Nutrition 0.000 claims description 3
- NFZYDZXHKFHPGA-UHFFFAOYSA-N 17alpha-hydroxygofruside Natural products C12CC(C)(C)CCC2(C(=O)OC2C(C(O)C(O)C(CO)O2)O)CCC(C2(CCC3C4(C)C)C)(C)C1=CCC2C3(C)CCC4OC1OC(C(O)=O)C(O)C(O)C1OC1OC(CO)C(O)C(O)C1O NFZYDZXHKFHPGA-UHFFFAOYSA-N 0.000 claims description 2
- QXQFFGOMXYKNBA-UHFFFAOYSA-N Chikusetsusaponin V Natural products CC1(C)CCC2(CCC3C(=CCC4C3(C)CCC5C(C)(C)C(CCC45C)OC6OC(C(O)C(O)C6OC7OC(CO)C(O)C(O)C7O)C(=O)O)C2C1)C(=O)OC8OC(CO)C(O)C(O)C8O QXQFFGOMXYKNBA-UHFFFAOYSA-N 0.000 claims description 2
- NFZYDZXHKFHPGA-QQHDHSITSA-N Chikusetsusaponin-V Chemical compound O([C@@H]1[C@@H](O)[C@H](O)[C@H](O[C@H]1O[C@H]1CC[C@]2(C)[C@H]3CC=C4[C@@]([C@@]3(CC[C@H]2C1(C)C)C)(C)CC[C@]1(CCC(C[C@H]14)(C)C)C(=O)O[C@H]1[C@@H]([C@@H](O)[C@H](O)[C@@H](CO)O1)O)C(O)=O)[C@@H]1O[C@H](CO)[C@@H](O)[C@H](O)[C@H]1O NFZYDZXHKFHPGA-QQHDHSITSA-N 0.000 claims description 2
- 230000000118 anti-neoplastic effect Effects 0.000 claims description 2
- 239000003480 eluent Substances 0.000 claims description 2
- 239000000945 filler Substances 0.000 claims description 2
- RBRANZURTULKJD-UHFFFAOYSA-N ginsenoside Ro Natural products CC1(C)CCC2(CCC3(C)C(=CCC4C5(C)CCC(OC6OC(C)(C(O)C(O)C6OC7OC(CO)C(O)C(O)C7O)C(=O)O)C(C)(C)C5CCC34C)C2C1)C(=O)OC8OC(CO)C(O)C(O)C8O RBRANZURTULKJD-UHFFFAOYSA-N 0.000 claims description 2
- 238000002156 mixing Methods 0.000 claims description 2
- 239000008213 purified water Substances 0.000 claims description 2
- 238000002604 ultrasonography Methods 0.000 claims description 2
- 238000004519 manufacturing process Methods 0.000 claims 2
- 206010028980 Neoplasm Diseases 0.000 abstract description 22
- 230000000694 effects Effects 0.000 abstract description 7
- 102000018711 Facilitative Glucose Transport Proteins Human genes 0.000 abstract description 3
- 108091052347 Glucose transporter family Proteins 0.000 abstract description 3
- 238000012986 modification Methods 0.000 abstract description 2
- 230000004048 modification Effects 0.000 abstract description 2
- 230000036571 hydration Effects 0.000 abstract 1
- 238000006703 hydration reaction Methods 0.000 abstract 1
- 210000004027 cell Anatomy 0.000 description 30
- 239000000243 solution Substances 0.000 description 28
- 239000000725 suspension Substances 0.000 description 14
- 239000000203 mixture Substances 0.000 description 13
- 241000699670 Mus sp. Species 0.000 description 11
- 210000004369 blood Anatomy 0.000 description 11
- 239000008280 blood Substances 0.000 description 11
- 238000005538 encapsulation Methods 0.000 description 11
- 239000006228 supernatant Substances 0.000 description 11
- -1 taxol compound Chemical class 0.000 description 11
- 238000009472 formulation Methods 0.000 description 9
- 229940108949 paclitaxel injection Drugs 0.000 description 8
- 238000005303 weighing Methods 0.000 description 8
- 239000002245 particle Substances 0.000 description 7
- 210000002381 plasma Anatomy 0.000 description 7
- GZYPWOGIYAIIPV-JBDTYSNRSA-N ginsenoside Rb1 Chemical compound C([C@H]1O[C@H]([C@@H]([C@@H](O)[C@@H]1O)O)O[C@@](C)(CCC=C(C)C)[C@@H]1[C@@H]2[C@@]([C@@]3(CC[C@H]4C(C)(C)[C@@H](O[C@H]5[C@@H]([C@@H](O)[C@H](O)[C@@H](CO)O5)O[C@H]5[C@@H]([C@@H](O)[C@H](O)[C@@H](CO)O5)O)CC[C@]4(C)[C@H]3C[C@H]2O)C)(C)CC1)O[C@@H]1O[C@H](CO)[C@@H](O)[C@H](O)[C@H]1O GZYPWOGIYAIIPV-JBDTYSNRSA-N 0.000 description 6
- 239000002609 medium Substances 0.000 description 6
- 230000004044 response Effects 0.000 description 6
- FBFMBWCLBGQEBU-RXMALORBSA-N (2s,3r,4s,5s,6r)-2-[(2r,3r,4s,5s,6r)-2-[[(3s,5r,6s,8r,9r,10r,12r,13r,14r,17s)-3,12-dihydroxy-4,4,8,10,14-pentamethyl-17-[(2s)-6-methyl-2-[(2s,3r,4s,5s,6r)-3,4,5-trihydroxy-6-(hydroxymethyl)oxan-2-yl]oxyhept-5-en-2-yl]-2,3,5,6,7,9,11,12,13,15,16,17-dodecah Chemical compound O([C@@](C)(CCC=C(C)C)[C@@H]1[C@@H]2[C@@]([C@@]3(C[C@@H]([C@H]4C(C)(C)[C@@H](O)CC[C@]4(C)[C@H]3C[C@H]2O)O[C@H]2[C@@H]([C@@H](O)[C@H](O)[C@@H](CO)O2)O[C@H]2[C@@H]([C@@H](O)[C@H](O)[C@@H](CO)O2)O)C)(C)CC1)[C@@H]1O[C@H](CO)[C@@H](O)[C@H](O)[C@H]1O FBFMBWCLBGQEBU-RXMALORBSA-N 0.000 description 5
- FBFMBWCLBGQEBU-GYMUUCMZSA-N 20-gluco-ginsenoside-Rf Natural products O([C@](CC/C=C(\C)/C)(C)[C@@H]1[C@H]2[C@H](O)C[C@H]3[C@](C)([C@]2(C)CC1)C[C@H](O[C@@H]1[C@H](O[C@H]2[C@H](O)[C@@H](O)[C@H](O)[C@@H](CO)O2)[C@@H](O)[C@H](O)[C@@H](CO)O1)[C@H]1C(C)(C)[C@@H](O)CC[C@]31C)[C@H]1[C@H](O)[C@@H](O)[C@H](O)[C@@H](CO)O1 FBFMBWCLBGQEBU-GYMUUCMZSA-N 0.000 description 5
- HYPFYJBWSTXDAS-UHFFFAOYSA-N Ginsenoside Rd Natural products CC(=CCCC(C)(OC1OC(CO)C(O)C(O)C1O)C2CCC3(C)C4CCC5C(C)(C)C(CCC5(C)C4CC(O)C23C)OC6OC(CO)C(O)C(O)C6OC7OC(CO)C(O)C(O)C7O)C HYPFYJBWSTXDAS-UHFFFAOYSA-N 0.000 description 5
- 208000002193 Pain Diseases 0.000 description 5
- 230000005540 biological transmission Effects 0.000 description 5
- 238000000338 in vitro Methods 0.000 description 5
- 229940090044 injection Drugs 0.000 description 5
- 239000007924 injection Substances 0.000 description 5
- 238000002347 injection Methods 0.000 description 5
- 239000012778 molding material Substances 0.000 description 5
- 238000011160 research Methods 0.000 description 5
- UOJAEODBOCLNBU-UHFFFAOYSA-N vinaginsenoside R4 Natural products C1CC(C2(CC(O)C3C(C)(C)C(OC4C(C(O)C(O)C(CO)O4)OC4C(C(O)C(O)C(CO)O4)O)CCC3(C)C2CC2O)C)(C)C2C1C(C)(CCC=C(C)C)OC1OC(CO)C(O)C(O)C1O UOJAEODBOCLNBU-UHFFFAOYSA-N 0.000 description 5
- 238000002474 experimental method Methods 0.000 description 4
- 238000011580 nude mouse model Methods 0.000 description 4
- 238000005406 washing Methods 0.000 description 4
- OKKJLVBELUTLKV-UHFFFAOYSA-N Methanol Chemical compound OC OKKJLVBELUTLKV-UHFFFAOYSA-N 0.000 description 3
- 238000004458 analytical method Methods 0.000 description 3
- 230000008901 benefit Effects 0.000 description 3
- 238000006243 chemical reaction Methods 0.000 description 3
- 238000010790 dilution Methods 0.000 description 3
- 239000012895 dilution Substances 0.000 description 3
- 235000019441 ethanol Nutrition 0.000 description 3
- 238000010438 heat treatment Methods 0.000 description 3
- 230000005764 inhibitory process Effects 0.000 description 3
- 238000011081 inoculation Methods 0.000 description 3
- 230000010189 intracellular transport Effects 0.000 description 3
- 239000007791 liquid phase Substances 0.000 description 3
- 238000005259 measurement Methods 0.000 description 3
- 238000001000 micrograph Methods 0.000 description 3
- 208000004296 neuralgia Diseases 0.000 description 3
- 208000021722 neuropathic pain Diseases 0.000 description 3
- 230000008569 process Effects 0.000 description 3
- 230000001737 promoting effect Effects 0.000 description 3
- 230000001988 toxicity Effects 0.000 description 3
- 231100000419 toxicity Toxicity 0.000 description 3
- 229930182493 triterpene saponin Natural products 0.000 description 3
- CKUVNOCSBYYHIS-UHFFFAOYSA-N (20R)-ginsenoside Rg3 Natural products CC(C)=CCCC(C)(O)C1CCC(C2(CCC3C4(C)C)C)(C)C1C(O)CC2C3(C)CCC4OC1OC(CO)C(O)C(O)C1O CKUVNOCSBYYHIS-UHFFFAOYSA-N 0.000 description 2
- CKUVNOCSBYYHIS-IRFFNABBSA-N (20S)-ginsenoside Rh2 Chemical compound O([C@H]1CC[C@]2(C)[C@H]3C[C@@H](O)[C@H]4[C@@]([C@@]3(CC[C@H]2C1(C)C)C)(C)CC[C@@H]4[C@@](C)(O)CCC=C(C)C)[C@@H]1O[C@H](CO)[C@@H](O)[C@H](O)[C@H]1O CKUVNOCSBYYHIS-IRFFNABBSA-N 0.000 description 2
- CKUVNOCSBYYHIS-LGYUXIIVSA-N 20(R)-Ginsenoside Rh2 Natural products O([C@H]1[C@H](O)[C@@H](O)[C@H](O)[C@@H](CO)O1)[C@@H]1C(C)(C)[C@H]2[C@@](C)([C@H]3[C@](C)([C@@]4(C)[C@H]([C@H](O)C3)[C@@H]([C@](O)(CC/C=C(\C)/C)C)CC4)CC2)CC1 CKUVNOCSBYYHIS-LGYUXIIVSA-N 0.000 description 2
- UFNDONGOJKNAES-UHFFFAOYSA-N Ginsenoside Rb1 Natural products CC(=CCCC(C)(OC1OC(COC2OC(CO)C(O)C(O)C2O)C(O)C(O)C1O)C3CCC4(C)C3C(O)CC5C6(C)CCC(OC7OC(CO)C(O)C(O)C7OC8OC(CO)C(O)C(O)C8O)C(C)(C)C6CC(O)C45C)C UFNDONGOJKNAES-UHFFFAOYSA-N 0.000 description 2
- 102000001554 Hemoglobins Human genes 0.000 description 2
- 108010054147 Hemoglobins Proteins 0.000 description 2
- 108010019160 Pancreatin Proteins 0.000 description 2
- 241000700159 Rattus Species 0.000 description 2
- 238000002835 absorbance Methods 0.000 description 2
- 210000001772 blood platelet Anatomy 0.000 description 2
- 238000009835 boiling Methods 0.000 description 2
- 239000013592 cell lysate Substances 0.000 description 2
- 239000006285 cell suspension Substances 0.000 description 2
- 239000004035 construction material Substances 0.000 description 2
- OORMXZNMRWBSTK-LGFJJATJSA-N dammarane Chemical compound C1CCC(C)(C)[C@@H]2CC[C@@]3(C)[C@]4(C)CC[C@H]([C@H](C)CCCC(C)C)[C@H]4CC[C@@H]3[C@]21C OORMXZNMRWBSTK-LGFJJATJSA-N 0.000 description 2
- 238000011161 development Methods 0.000 description 2
- 230000018109 developmental process Effects 0.000 description 2
- 230000002708 enhancing effect Effects 0.000 description 2
- 238000011156 evaluation Methods 0.000 description 2
- 238000001704 evaporation Methods 0.000 description 2
- 230000037406 food intake Effects 0.000 description 2
- TXEWRVNOAJOINC-UHFFFAOYSA-N ginsenoside Rb2 Natural products CC(=CCCC(OC1OC(COC2OCC(O)C(O)C2O)C(O)C(O)C1O)C3CCC4(C)C3C(O)CC5C6(C)CCC(OC7OC(CO)C(O)C(O)C7OC8OC(CO)C(O)C(O)C8O)C(C)(C)C6CCC45C)C TXEWRVNOAJOINC-UHFFFAOYSA-N 0.000 description 2
- 239000001963 growth medium Substances 0.000 description 2
- 230000007062 hydrolysis Effects 0.000 description 2
- 238000006460 hydrolysis reaction Methods 0.000 description 2
- 230000001939 inductive effect Effects 0.000 description 2
- 210000002490 intestinal epithelial cell Anatomy 0.000 description 2
- 210000000265 leukocyte Anatomy 0.000 description 2
- 150000002772 monosaccharides Chemical class 0.000 description 2
- 210000000440 neutrophil Anatomy 0.000 description 2
- 208000002154 non-small cell lung carcinoma Diseases 0.000 description 2
- 229940055695 pancreatin Drugs 0.000 description 2
- 239000008188 pellet Substances 0.000 description 2
- 229920000642 polymer Polymers 0.000 description 2
- 230000035755 proliferation Effects 0.000 description 2
- 238000007920 subcutaneous administration Methods 0.000 description 2
- 239000000126 substance Substances 0.000 description 2
- 230000001225 therapeutic effect Effects 0.000 description 2
- 230000031998 transcytosis Effects 0.000 description 2
- 208000029729 tumor suppressor gene on chromosome 11 Diseases 0.000 description 2
- PYXFVCFISTUSOO-HKUCOEKDSA-N (20S)-protopanaxadiol Chemical compound C1C[C@H](O)C(C)(C)[C@@H]2CC[C@@]3(C)[C@]4(C)CC[C@H]([C@@](C)(O)CCC=C(C)C)[C@H]4[C@H](O)C[C@@H]3[C@]21C PYXFVCFISTUSOO-HKUCOEKDSA-N 0.000 description 1
- MIJYXULNPSFWEK-GTOFXWBISA-N 3beta-hydroxyolean-12-en-28-oic acid Chemical compound C1C[C@H](O)C(C)(C)[C@@H]2CC[C@@]3(C)[C@]4(C)CC[C@@]5(C(O)=O)CCC(C)(C)C[C@H]5C4=CC[C@@H]3[C@]21C MIJYXULNPSFWEK-GTOFXWBISA-N 0.000 description 1
- 206010067484 Adverse reaction Diseases 0.000 description 1
- TWCMVXMQHSVIOJ-UHFFFAOYSA-N Aglycone of yadanzioside D Natural products COC(=O)C12OCC34C(CC5C(=CC(O)C(O)C5(C)C3C(O)C1O)C)OC(=O)C(OC(=O)C)C24 TWCMVXMQHSVIOJ-UHFFFAOYSA-N 0.000 description 1
- 108010012934 Albumin-Bound Paclitaxel Proteins 0.000 description 1
- 108010088751 Albumins Proteins 0.000 description 1
- 102000009027 Albumins Human genes 0.000 description 1
- PLMKQQMDOMTZGG-UHFFFAOYSA-N Astrantiagenin E-methylester Natural products CC12CCC(O)C(C)(CO)C1CCC1(C)C2CC=C2C3CC(C)(C)CCC3(C(=O)OC)CCC21C PLMKQQMDOMTZGG-UHFFFAOYSA-N 0.000 description 1
- 108091003079 Bovine Serum Albumin Proteins 0.000 description 1
- 206010010904 Convulsion Diseases 0.000 description 1
- 239000006144 Dulbecco’s modified Eagle's medium Substances 0.000 description 1
- JKLISIRFYWXLQG-UHFFFAOYSA-N Epioleonolsaeure Natural products C1CC(O)C(C)(C)C2CCC3(C)C4(C)CCC5(C(O)=O)CCC(C)(C)CC5C4CCC3C21C JKLISIRFYWXLQG-UHFFFAOYSA-N 0.000 description 1
- 238000012449 Kunming mouse Methods 0.000 description 1
- 241001465754 Metazoa Species 0.000 description 1
- 241000699660 Mus musculus Species 0.000 description 1
- YBRJHZPWOMJYKQ-UHFFFAOYSA-N Oleanolic acid Natural products CC1(C)CC2C3=CCC4C5(C)CCC(O)C(C)(C)C5CCC4(C)C3(C)CCC2(C1)C(=O)O YBRJHZPWOMJYKQ-UHFFFAOYSA-N 0.000 description 1
- MIJYXULNPSFWEK-UHFFFAOYSA-N Oleanolinsaeure Natural products C1CC(O)C(C)(C)C2CCC3(C)C4(C)CCC5(C(O)=O)CCC(C)(C)CC5C4=CCC3C21C MIJYXULNPSFWEK-UHFFFAOYSA-N 0.000 description 1
- 229940123237 Taxane Drugs 0.000 description 1
- YURJSTAIMNSZAE-UHFFFAOYSA-N UNPD89172 Natural products C1CC(C2(CC(C3C(C)(C)C(O)CCC3(C)C2CC2O)OC3C(C(O)C(O)C(CO)O3)O)C)(C)C2C1C(C)(CCC=C(C)C)OC1OC(CO)C(O)C(O)C1O YURJSTAIMNSZAE-UHFFFAOYSA-N 0.000 description 1
- 208000027418 Wounds and injury Diseases 0.000 description 1
- 230000002159 abnormal effect Effects 0.000 description 1
- 229940028652 abraxane Drugs 0.000 description 1
- 230000006838 adverse reaction Effects 0.000 description 1
- 150000001413 amino acids Chemical class 0.000 description 1
- 230000003698 anagen phase Effects 0.000 description 1
- 239000003146 anticoagulant agent Substances 0.000 description 1
- 229940127219 anticoagulant drug Drugs 0.000 description 1
- 230000006907 apoptotic process Effects 0.000 description 1
- PYXFVCFISTUSOO-UHFFFAOYSA-N betulafolienetriol Natural products C1CC(O)C(C)(C)C2CCC3(C)C4(C)CCC(C(C)(O)CCC=C(C)C)C4C(O)CC3C21C PYXFVCFISTUSOO-UHFFFAOYSA-N 0.000 description 1
- 230000015572 biosynthetic process Effects 0.000 description 1
- 230000037396 body weight Effects 0.000 description 1
- 210000005252 bulbus oculi Anatomy 0.000 description 1
- 150000001720 carbohydrates Chemical class 0.000 description 1
- 238000010609 cell counting kit-8 assay Methods 0.000 description 1
- 230000024245 cell differentiation Effects 0.000 description 1
- 230000008859 change Effects 0.000 description 1
- 239000003153 chemical reaction reagent Substances 0.000 description 1
- 201000010897 colon adenocarcinoma Diseases 0.000 description 1
- 208000029742 colonic neoplasm Diseases 0.000 description 1
- 239000011365 complex material Substances 0.000 description 1
- 150000001875 compounds Chemical class 0.000 description 1
- 238000007796 conventional method Methods 0.000 description 1
- 230000036461 convulsion Effects 0.000 description 1
- 231100000433 cytotoxic Toxicity 0.000 description 1
- 230000001472 cytotoxic effect Effects 0.000 description 1
- 230000003013 cytotoxicity Effects 0.000 description 1
- 231100000135 cytotoxicity Toxicity 0.000 description 1
- 230000006378 damage Effects 0.000 description 1
- 230000013872 defecation Effects 0.000 description 1
- 230000007547 defect Effects 0.000 description 1
- 230000003111 delayed effect Effects 0.000 description 1
- 238000001514 detection method Methods 0.000 description 1
- 238000010586 diagram Methods 0.000 description 1
- 239000002552 dosage form Substances 0.000 description 1
- 231100000371 dose-limiting toxicity Toxicity 0.000 description 1
- 230000035622 drinking Effects 0.000 description 1
- 238000012377 drug delivery Methods 0.000 description 1
- 230000008451 emotion Effects 0.000 description 1
- 230000012202 endocytosis Effects 0.000 description 1
- 230000035611 feeding Effects 0.000 description 1
- 239000012091 fetal bovine serum Substances 0.000 description 1
- 238000007710 freezing Methods 0.000 description 1
- 230000008014 freezing Effects 0.000 description 1
- 230000006870 function Effects 0.000 description 1
- PWAOOJDMFUQOKB-WCZZMFLVSA-N ginsenoside Re Chemical compound O[C@@H]1[C@H](O)[C@@H](O)[C@H](C)O[C@H]1O[C@H]1[C@H](O[C@@H]2[C@H]3C(C)(C)[C@@H](O)CC[C@]3(C)[C@@H]3[C@@]([C@@]4(CC[C@@H]([C@H]4[C@H](O)C3)[C@](C)(CCC=C(C)C)O[C@H]3[C@@H]([C@@H](O)[C@H](O)[C@@H](CO)O3)O)C)(C)C2)O[C@H](CO)[C@@H](O)[C@@H]1O PWAOOJDMFUQOKB-WCZZMFLVSA-N 0.000 description 1
- YURJSTAIMNSZAE-HHNZYBFYSA-N ginsenoside Rg1 Chemical compound O([C@@](C)(CCC=C(C)C)[C@@H]1[C@@H]2[C@@]([C@@]3(C[C@@H]([C@H]4C(C)(C)[C@@H](O)CC[C@]4(C)[C@H]3C[C@H]2O)O[C@H]2[C@@H]([C@@H](O)[C@H](O)[C@@H](CO)O2)O)C)(C)CC1)[C@@H]1O[C@H](CO)[C@@H](O)[C@H](O)[C@H]1O YURJSTAIMNSZAE-HHNZYBFYSA-N 0.000 description 1
- CBEHEBUBNAGGKC-UHFFFAOYSA-N ginsenoside Rg1 Natural products CC(=CCCC(C)(OC1OC(CO)C(O)C(O)C1O)C2CCC3(C)C2C(O)CC4C5(C)CCC(O)C(C)(C)C5CC(OC6OC(CO)C(O)C(O)C6O)C34C)C CBEHEBUBNAGGKC-UHFFFAOYSA-N 0.000 description 1
- AOGZLQUEBLOQCI-UHFFFAOYSA-N ginsenoside-Re Natural products CC1OC(OCC2OC(OC3CC4(C)C(CC(O)C5C(CCC45C)C(C)(CCC=C(C)C)OC6OC(CO)C(O)C(O)C6O)C7(C)CCC(O)C(C)(C)C37)C(O)C(O)C2O)C(O)C(O)C1O AOGZLQUEBLOQCI-UHFFFAOYSA-N 0.000 description 1
- 230000006377 glucose transport Effects 0.000 description 1
- PFOARMALXZGCHY-UHFFFAOYSA-N homoegonol Natural products C1=C(OC)C(OC)=CC=C1C1=CC2=CC(CCCO)=CC(OC)=C2O1 PFOARMALXZGCHY-UHFFFAOYSA-N 0.000 description 1
- 230000000887 hydrating effect Effects 0.000 description 1
- 239000000413 hydrolysate Substances 0.000 description 1
- 230000036039 immunity Effects 0.000 description 1
- 238000001727 in vivo Methods 0.000 description 1
- 238000011534 incubation Methods 0.000 description 1
- 230000002401 inhibitory effect Effects 0.000 description 1
- 208000014674 injury Diseases 0.000 description 1
- 230000009191 jumping Effects 0.000 description 1
- 238000011031 large-scale manufacturing process Methods 0.000 description 1
- 230000027939 micturition Effects 0.000 description 1
- 238000012544 monitoring process Methods 0.000 description 1
- 239000002105 nanoparticle Substances 0.000 description 1
- 208000004235 neutropenia Diseases 0.000 description 1
- 229940100243 oleanolic acid Drugs 0.000 description 1
- 239000006186 oral dosage form Substances 0.000 description 1
- 238000012856 packing Methods 0.000 description 1
- RCINICONZNJXQF-QWZZYFJGSA-N paclitaxel Polymers O([C@H]1[C@@]2(C[C@@H](C(C)=C(C2(C)C)[C@H](C([C@]2(C)[C@H](O)C[C@H]3OC[C@]3(C21)OC(C)=O)=O)OC(=O)C)OC(=O)[C@@H](O)[C@H](NC(=O)C=1C=CC=CC=1)C=1C=CC=CC=1)O)C(=O)C1=CC=CC=C1 RCINICONZNJXQF-QWZZYFJGSA-N 0.000 description 1
- 230000035699 permeability Effects 0.000 description 1
- 239000000825 pharmaceutical preparation Substances 0.000 description 1
- OQUKIQWCVTZJAF-UHFFFAOYSA-N phenol;sulfuric acid Chemical compound OS(O)(=O)=O.OC1=CC=CC=C1 OQUKIQWCVTZJAF-UHFFFAOYSA-N 0.000 description 1
- 239000002504 physiological saline solution Substances 0.000 description 1
- 210000003240 portal vein Anatomy 0.000 description 1
- 239000000047 product Substances 0.000 description 1
- HZLWUYJLOIAQFC-UHFFFAOYSA-N prosapogenin PS-A Natural products C12CC(C)(C)CCC2(C(O)=O)CCC(C2(CCC3C4(C)C)C)(C)C1=CCC2C3(C)CCC4OC1OCC(O)C(O)C1O HZLWUYJLOIAQFC-UHFFFAOYSA-N 0.000 description 1
- SWQINCWATANGKN-UHFFFAOYSA-N protopanaxadiol Natural products CC(CCC=C(C)C)C1CCC2(C)C1C(O)CC1C3(C)CCC(O)C(C)(C)C3CCC21C SWQINCWATANGKN-UHFFFAOYSA-N 0.000 description 1
- SHCBCKBYTHZQGZ-DLHMIPLTSA-N protopanaxatriol Chemical compound C1C[C@H](O)C(C)(C)[C@@H]2[C@@H](O)C[C@@]3(C)[C@]4(C)CC[C@H]([C@](C)(O)CCC=C(C)C)[C@H]4[C@H](O)C[C@@H]3[C@]21C SHCBCKBYTHZQGZ-DLHMIPLTSA-N 0.000 description 1
- BBEUDPAEKGPXDG-UHFFFAOYSA-N protopanaxatriol Natural products CC(CCC=C(C)C)C1CCC2(C)C1C(O)CC3C4(C)CCC(O)C(C)(C)C4C(O)CC23C BBEUDPAEKGPXDG-UHFFFAOYSA-N 0.000 description 1
- 239000001397 quillaja saponaria molina bark Substances 0.000 description 1
- 230000035484 reaction time Effects 0.000 description 1
- 238000013214 routine measurement Methods 0.000 description 1
- 229930182490 saponin Natural products 0.000 description 1
- 150000007949 saponins Chemical class 0.000 description 1
- 235000017709 saponins Nutrition 0.000 description 1
- 238000007789 sealing Methods 0.000 description 1
- 239000012679 serum free medium Substances 0.000 description 1
- 239000007787 solid Substances 0.000 description 1
- 239000007858 starting material Substances 0.000 description 1
- 238000005728 strengthening Methods 0.000 description 1
- 239000000758 substrate Substances 0.000 description 1
- 238000003786 synthesis reaction Methods 0.000 description 1
- 229920002994 synthetic fiber Polymers 0.000 description 1
- 125000002456 taxol group Polymers 0.000 description 1
- 238000012360 testing method Methods 0.000 description 1
- WROMPOXWARCANT-UHFFFAOYSA-N tfa trifluoroacetic acid Chemical compound OC(=O)C(F)(F)F.OC(=O)C(F)(F)F WROMPOXWARCANT-UHFFFAOYSA-N 0.000 description 1
- 238000002560 therapeutic procedure Methods 0.000 description 1
- 235000013619 trace mineral Nutrition 0.000 description 1
- 239000011573 trace mineral Substances 0.000 description 1
- 230000032258 transport Effects 0.000 description 1
- 210000004881 tumor cell Anatomy 0.000 description 1
- 230000004565 tumor cell growth Effects 0.000 description 1
- 230000004614 tumor growth Effects 0.000 description 1
- 238000001291 vacuum drying Methods 0.000 description 1
- 239000000341 volatile oil Substances 0.000 description 1
- 230000004584 weight gain Effects 0.000 description 1
- 235000019786 weight gain Nutrition 0.000 description 1
- 230000004580 weight loss Effects 0.000 description 1
Abstract
The invention discloses an oral micelle taking ginseng active ingredients as auxiliary materials, a preparation method and application thereof, wherein ginsenoside is taken as a forming material of the micelle, ginseng polysaccharide is taken as a modification material of the micelle, paclitaxel or docetaxel is taken as a model drug, and the oral micelle taking the ginsenoside as the forming material is prepared by a film hydration method. On one hand, the micelle constructed by the invention can promote the drug transmembrane property and the ginseng polysaccharide property by utilizing the glucose transporter property by the micelle, so that the oral absorption of the insoluble drug taxol and docetaxel is improved; on the one hand, the antitumor effect of the active ingredients of the ginseng is assisted by the ginsenoside and the ginseng polysaccharide, so that the tumor treatment effect of the taxol and the docetaxel is improved.
Description
Technical Field
The invention belongs to the technical field of pharmaceutical preparations, and particularly relates to an oral micelle taking ginseng active ingredients as auxiliary materials, and a preparation method and application thereof.
Background
Paclitaxel (Paclitaxel) and docetaxel (docetaxel), both of which are cytotoxic taxanes that are widely used clinically in tumor therapy. However, both are typical BCS class iv compounds, which have poor solubility and permeability, resulting in very low oral bioavailability (around 1%). The existing preparation containing paclitaxel can be paclitaxel injection (Taxol), paclitaxel albumin nanoparticle for injection (Abraxane) and paclitaxel polymer micelle for injection (Genexol-PM); the preparation containing docetaxel mainly comprises docetaxel injection (Taxote) and the like; all are clinical large varieties. However, due to poor oral absorption, the varieties are injection administration forms, and have the defects of poor patient compliance and more adverse reactions, if the varieties are developed into oral forms, the clinical application of the varieties containing taxol or docetaxel is facilitated.
The key point of developing taxol and docetaxel oral preparation is that whether the oral absorption of taxol and docetaxel can be improved on the premise of safe use; among the numerous formulation techniques, micelles are a common technique. When the concentration of the amphiphilic substance in the medium exceeds the critical micelle concentration, the amphiphilic substance can spontaneously form micelles, so that the preparation process of the micelles is relatively simple and convenient compared with the common nano preparation. However, since the therapeutic effect of the drug is difficult to be further exerted by the effect of the micelle alone, materials with different functions are required to be added into the micelle construction material, so that the preparation process of the micelle construction material is relatively complex, the problem of safety is easy to cause, and the clinical development value of the polymer micelle is restricted to limit the development of a micelle drug delivery system. In addition, the micelle which is marketed at present is an injection administration dosage form, and no oral dosage form is marketed yet. The main reason for limiting the marketing of oral micelles is that oral micelles cannot greatly improve the oral bioavailability of poorly soluble drugs. At present, research shows that the micelle has the characteristic of easy ingestion and difficult transcytosis, namely, the micelle can be easily ingested by intestinal epithelial cells, but is difficult to transport to the basal membrane side in cells, so that the micelle is influenced to be released out of the cells through endocytosis, and is absorbed by portal veins and then enters whole-body blood.
Ginseng is a medicine with abundant use experience in China, and contains various components such as saponin, polysaccharide, volatile oil, protein, amino acid, trace elements and the like, wherein the ginsenoside and the ginseng polysaccharide are the main active components of the ginseng. According to the difference of aglycone structures, ginsenosides can be classified into dammarane type tetracyclic triterpene saponins, oleanolic acid type pentacyclic triterpene saponins (Ro, ri, rh3, F4, etc.), and octoelevator type ginsenosides (F11, RT5, RT4, etc.) 3 types, wherein dammarane type tetracyclic triterpene saponins are further classified into protopanaxadiol (Rb 1, rb2, rb3, rc, rd, rh2, etc.) and protopanaxatriol (Re, rg1, rg2, rf, rh1, etc.) 2 types. In the research process of the invention, we find that various ginsenoside components have certain capacity of spontaneously forming micelles; in addition, in the course of our research, we found that the traditional Chinese medicine polysaccharide can promote the oral absorption of the medicine to a certain extent, particularly because many saccharide components in the polysaccharide are substrates of glucose transporter, which can promote the intracellular transport of the medicine, thereby improving the oral absorption of the medicine.
Disclosure of Invention
The invention aims to safely and effectively improve the oral absorption of indissolvable drugs taxol and docetaxel, thereby providing a preparation method and application of an oral micelle taking ginseng active ingredients as auxiliary materials for enhancing the anti-tumor effect of taxol and docetaxel.
Based on the background and the findings in the research process, the invention adopts the natural ginsenoside as the forming material of the micelle, and plays the effect of promoting the oral absorption of the drug by the micelle; the affinity of ginseng polysaccharide and glucose transporter is utilized to improve the intracellular transport efficiency of the micelle in intestinal epithelial cells, so that the effect of promoting the oral absorption of the drug of the micelle is further improved; in addition, ginsenoside and ginseng polysaccharide have auxiliary anti-tumor effect (such as enhancing immunity (strengthening body resistance), inducing tumor cell differentiation and apoptosis, and inhibiting tumor cell growth), and can further improve anti-tumor effect of paclitaxel and docetaxel. The oral micelle only adopts natural ginsenoside and ginseng polysaccharide as the assembly materials of the micelle, and has better in vivo use safety compared with the synthetic materials.
The invention aims at realizing the following technical scheme:
an oral micelle taking ginseng active ingredients as auxiliary materials is characterized in that: the oral micelle comprises auxiliary materials and an anti-tumor indissoluble drug, wherein the auxiliary materials are ginsenoside, and the anti-tumor indissoluble drug is a taxol compound.
Preferably, the mass ratio of the anti-tumor indissoluble medicine to the ginsenoside is 1 (0.2-30), more preferably 1 (0.2-9), and even more preferably 1 (4-9).
Preferably, the taxol compound is taxol or docetaxel.
Preferably, the ginsenoside is one or more of ginsenoside Ro, ri, rh3, F4, F11, RT5, RT4, rb1, rb2, rb3, rc, rd, rh2, re, rg1, rg2, rf, PPD, rh1, and the like; more preferably, the composition contains one or more of ginsenoside Rb1, rd, rh2, re, rg1 and PPD; more preferred is one or more of ginsenoside Rb1, rd and Rh 2.
Preferably, the oral micelle further comprises a modified material ginseng polysaccharide, wherein the mass ratio of the ginseng polysaccharide to the ginsenoside is 1 (0.05-10), more preferably 1 (0.2-5).
Preferably, the extraction method of ginseng polysaccharide comprises the following steps:
a. adding pure water into ginseng sample according to a feed liquid ratio of 1:30, extracting for 0.5-1h, cooling, centrifuging, precipitating with ethanol, filtering, collecting precipitate, and drying to obtain ginseng crude polysaccharide;
b. and d, performing enzymolysis by adopting alpha-amylase to remove starch in the ginseng crude polysaccharide obtained in the step a, wherein the enzymolysis conditions are as follows: 800 U alpha-amylase, and carrying out enzymolysis for 0.5-1h at 50-60 ℃;
c. and d, carrying out enzymolysis by papain to remove protein in the ginseng crude polysaccharide obtained in the step b, wherein the enzymolysis conditions are as follows: 200 U papain is subjected to enzymolysis at 30-40 ℃ for 1-3 h to obtain crude ginseng polysaccharide after enzymolysis;
d. preparing crude ginseng polysaccharide after enzymolysis, performing column chromatography elution by using DEAE-52 cellulose filler, eluting with distilled water, 0.1, 0.2, 0.3, 0.4, 0.5, 0.6, 0.7 and 0.8mol/L NaCl solution respectively in sequence, eluting at the flow rate of 1mL/min, collecting eluent to obtain polysaccharide 1, 2, 3, 4, 5, 6, 7, 8 and 9 respectively, and obtaining polysaccharide 2 to obtain the ginseng polysaccharide.
The invention also provides a preparation method of the oral micelle taking the ginseng active ingredient as an auxiliary material, which comprises the following steps:
(1) Mixing ginsenoside and antineoplastic insoluble medicine, placing in absolute ethanol, and dissolving by ultrasound to obtain medicinal and ginsenoside-containing solution;
(2) Placing the solution containing the medicine and the ginsenoside obtained in the step (1) into a rotary evaporator (or similar principle equipment), and removing ethanol by rotary evaporation at the temperature of 30-60 ℃ to enable the solution to slowly form a uniform film;
(3) Adding purified water or ginseng polysaccharide water solution into the film obtained in the step (2), carrying out ultrasonic treatment at 20-55 ℃ for 0.5-15 min, more preferably ultrasonic treatment at 5-15 min, completely hydrating the film, carrying out constant-temperature vibration at 30-40 ℃ for 0.5-2 h, more preferably vibration for 0.5-1h, and then filtering with a 0.22 mu m filter membrane to obtain paclitaxel ginsenoside micelle or ginseng polysaccharide modified paclitaxel ginsenoside micelle.
Preferably, in the step (1), the ultrasonic time is 10 to 40min, more preferably 15 to 30min; in the solution containing the medicine and the ginsenoside, the concentration of the anti-tumor indissoluble medicine is 0.01-20 mg/mL, and more preferably 0.1-0.6 mg/mL. In the step (2), the time of the rotary evaporation is 5 to 60 minutes, more preferably 15 to 30 minutes.
The invention also provides an application of the oral micelle taking the ginseng active ingredient as an auxiliary material in preparing an anti-tumor medicament.
The invention also provides an application of the oral micelle taking the ginseng active ingredient as an auxiliary material in improving the oral absorption of paclitaxel or docetaxel.
The invention has the advantages that: when the micelle is prepared, natural ginseng active ingredients are selected as auxiliary materials, so that the problems of poor safety, complex material synthesis steps and the like of the existing micelle forming materials can be solved. The ginsenoside is selected as a material to prepare the micelle, the capacity of promoting the oral absorption of the drug of the micelle can be exerted, and the ginseng polysaccharide is selected as a modification material to realize the efficient intracellular transport of the micelle by means of a glucose transport way, so that the problem that the oral absorption of the micelle is influenced due to easy ingestion and difficult transcytosis of the existing micelle is solved; therefore, the prepared micelle can better improve the oral absorption of the medicine. In addition, the antitumor effect of ginsenoside and ginseng polysaccharide can further improve the antitumor effect of paclitaxel (or docetaxel). The oral micelle disclosed by the invention has the advantages of high safety, good oral absorption, high anti-tumor effect and the like. The preparation equipment of the oral micelle is common equipment of pharmaceutical enterprises, has mild preparation conditions, is simple and controllable, is suitable for large-scale production, and has higher popularization and application values.
Drawings
FIG. 1 is a Transmission Electron Microscope (TEM) image of paclitaxel ginsenoside micelle and ginseng polysaccharide modified paclitaxel ginsenoside micelle according to the present invention (wherein A, B is paclitaxel ginsenoside micelle prepared in examples 3 and 4, respectively; C, D is paclitaxel ginsenoside micelle modified by ginseng polysaccharide prepared in examples 8 and 10, respectively);
FIG. 2 shows the results of in vitro uptake of polysaccharide and paclitaxel suspensions of the present invention by caco-2 cells.
FIG. 3 is a schematic of the in vitro toxicity of various paclitaxel formulations of the present invention to A549 cells;
FIG. 4 shows the results of in vitro uptake of various paclitaxel formulations of the present invention by A549 cells;
FIG. 5 is a graph of paclitaxel plasma drug concentration versus time according to the present invention;
FIG. 6 is a graph of plasma drug concentration versus time for docetaxel according to the present invention;
FIG. 7 is a graph showing the thermal pain response latency profile of various groups of paclitaxel formulations according to the present invention;
FIG. 8 shows the blood routine measurement results (10 mg/kg) of each paclitaxel formulation of the present invention.
Detailed Description
In order to further understand the present invention, a technical solution in the embodiments of the present invention will be clearly and completely described below in conjunction with the embodiments of the present invention, and it is apparent that the described embodiments are only some embodiments of the present invention, but not all embodiments. All other embodiments, which can be made by those skilled in the art based on the embodiments of the invention without making any inventive effort, are intended to be within the scope of the invention.
The experimental methods, in which specific conditions are not noted in the following examples, were selected according to conventional methods and conditions, or according to the commercial specifications.
Example 1
The preparation method of paclitaxel ginsenoside micelle comprises the following steps:
a. weighing 20mg of ginsenoside, 5mg of taxol in a round bottom flask, adding 15mL of absolute ethyl alcohol, and performing ultrasonic treatment for 15min (25 ℃ and 40 kHz) to completely dissolve the ginsenoside.
b. The solution obtained in step a was subjected to rotary evaporation (vacuum: 90-110 par) at 40℃for 15min to form a uniform film.
c. Adding 10mL of pure water into the film obtained in the step b, carrying out ultrasonic treatment for 15min (25 ℃ and 40 kHz) to hydrate the film, oscillating for 1h at a constant temperature of 40 ℃, and filtering the obtained liquid with a 0.22um filter membrane to obtain the taxol ginsenoside micelle. The obtained micelle had a particle diameter of 227.+ -. 36nm and a PDI of 0.189.+ -. 0.019.
According to the method, one of the ginsenoside in the following table 1 is selected as an auxiliary material, and the encapsulation rate of the obtained micelle is shown in the following table 1:
TABLE 1 encapsulation efficiency of micelles with single ginsenoside as molding material
Encapsulation efficiency = mass of drug contained in micelle/amount of drug added
Example 2
The preparation method of paclitaxel ginsenoside micelle comprises the following steps: 30mg of ginsenoside, 1mg of paclitaxel and the rest of steps are the same as in example 1. The obtained micelle size was 248.+ -. 41nm, and the PDI was 0.195.+ -. 0.023.
According to the method, one of the ginsenoside in the following table 2 is selected as an auxiliary material, and the encapsulation rate of the obtained micelle is shown in the following table 2:
TABLE 2 encapsulation efficiency of micelles with single ginsenoside as molding material
Example 3
The preparation method of paclitaxel ginsenoside micelle comprises the following steps:
a. weighing 4.5mg of each of the two ginsenosides, 1mg of paclitaxel in a round bottom flask, adding 10mL of absolute ethanol, and performing ultrasonic treatment for 30min (25 ℃ C., 40 kHz) to completely dissolve the ginsenosides.
b. And c, performing rotary evaporation (vacuum degree: 90-110 par) on the solution obtained in the step a at 30 ℃ for 30min to form a uniform film.
c. Adding 10mL of pure water into the film obtained in the step b, carrying out ultrasonic treatment for 5min (25 ℃ and 40 kHz) to hydrate the film, oscillating for 1h at a constant temperature of 30 ℃, and passing the obtained liquid through a 0.22um filter membrane to obtain the taxol ginsenoside micelle, wherein a transmission electron microscope image of the taxol ginsenoside micelle is shown in figure 1. The obtained micelle size was 158.+ -. 21nm and PDI was 0.166.+ -. 0.054.
According to the method, two different combinations of ginsenoside in the following table 3 are selected as auxiliary materials respectively, and the encapsulation rate of the obtained micelle is shown in the following table 3:
TABLE 3 encapsulation efficiency of micelles with two ginsenosides as molding materials
Example 4
The preparation method of paclitaxel ginsenoside micelle comprises the following steps: 10mg of each of the two ginsenosides, 1mg of paclitaxel, were weighed into a round bottom flask and the rest of the procedure was as in example 3. The transmission electron microscope diagram is shown in figure 1. The obtained micelle had a particle size of 155.+ -. 19nm and a PDI of 0.155.+ -. 0.081.
According to the method, two different combinations of the ginsenoside in the following table 4 are selected as auxiliary materials respectively, and the encapsulation rate of the obtained micelle is shown in the following table 4:
table 4 encapsulation efficiency of micelles with two ginsenosides as molding materials
Example 5
The preparation method of the docetaxel ginsenoside micelle comprises the following steps: two ginsenosides each 4.5mg, docetaxel 1mg were weighed in round bottom flask and the rest of the procedure was as in example 2. The obtained micelle has a particle size of 137+ -14 nm and PDI of 0.192+ -0.021.
According to the method, two different combinations of the ginsenoside in the following table 5 are selected as auxiliary materials respectively, and the encapsulation rate of the obtained micelle is shown in the following table 5:
table 5 encapsulation efficiency of micelles with two ginsenosides as molding materials
Example 6
Ginseng polysaccharide extraction method
a. Weighing a proper amount of ginseng sample in a beaker, and adding pure water according to a feed liquid ratio of 1:30 (g/mL). Ultrasonic extracting at 70deg.C and 60 Hz for 30min, cooling, centrifuging at 6000 r/min, adding 4 times volume of absolute ethanol into supernatant, and precipitating with ethanol at-20deg.C for 24 h. Filtering, collecting precipitate, and drying in vacuum oven (70deg.C) to constant weight to obtain crude Ginseng radix polysaccharide (GSP).
b. The starch of the above extracted GPS was removed using alpha-amylase. The GPS solution of 10mg/mL was carefully configured. Adding 800U alpha-amylase into the prepared GPS solution, performing enzymolysis for 30min at 50-60 ℃, transferring into a boiling water bath, and heating for 5min to terminate the enzymolysis reaction. After the solution was cooled to room temperature, it was centrifuged (4000 r/min,10 min) and the supernatant was collected.
c. Papain is used to remove proteins from the extracted GPS. 200U papain is added into the solution for enzymolysis at 30-40 ℃ for 2h, and then the solution is transferred into a boiling water bath for heating for 5min to terminate the enzymolysis reaction. After the solution was cooled to room temperature, it was centrifuged (4000 r/min,10 min) and the supernatant was collected. Heating the supernatant to 70 ℃ to evaporate excessive water, concentrating to 50 mL, adding 200 mL absolute ethyl alcohol into the concentrated solution, shaking uniformly, freezing in a refrigerator at-20 ℃ and standing for 24 h, filtering to obtain white solid, and drying in a vacuum drying oven (60 ℃) to constant weight to obtain the GPS after enzymolysis of 4.8 g.
d. Taking GPS 4.0 g after enzymolysis, and preparing 10mg/mL solution. The solution was eluted by column chromatography using DEAE-52 cellulose packing, and eluted with distilled water, 0.1, 0.2, 0.3, 0.4, 0.5, 0.6, 0.7, 0.8mol/L NaCl solution, respectively, at a flow rate of 1mL/min, collected using test tubes, and 10mL per tube. The absorbance of each bottle of liquid was measured by the phenol-sulfuric acid method, an elution curve was drawn, the solution having an absorption peak was collected, and the collected solution was freeze-dried (-40 ℃). 9 polysaccharides were obtained in total.
e. Accurately weighing 2mg of purified ginseng polysaccharide sample, placing the sample in a reaction kettle, adding 2mL of 2mol/L trifluoroacetic acid (TFA) solution, sealing, placing the solution in a 117 ℃ oven for hydrolysis 5h, taking supernatant, evaporating the supernatant to dryness, repeatedly washing the supernatant with methanol for 3 times, and evaporating the hydrolysate to dryness. The hydrolysis products were subjected to liquid phase analysis to calculate the amount of each monosaccharide. The results are shown in Table 6 below:
TABLE 6 yields of different ginseng polysaccharides and representative monosaccharide ratios
Example 7
Dilution of the suspension of caco-2 cells (human cloned colon adenocarcinoma cells) to a concentration of 2X 10 5 Plates were plated at 24 well plates at 500 μl/well and placed in cell incubators for 24 h. The cells were removed and then washed 2 times with PBS. The paclitaxel suspension was diluted to 20. Mu.g/mL with high sugar medium, and each well was added with the diluted paclitaxel suspension and 200. Mu.g/mL of 9 polysaccharide solutions, respectively. Incubating in incubator for 2h, discarding the liquid medicine and washing the cells with PBS for 2 times; cells were digested by adding 100. Mu.L of pancreatin, disrupted by adding 200. Mu.L of cell lysate, and centrifuged at 12000r/min for 10min. The supernatant was analyzed for drug concentration using a high performance liquid phase, and the lower protein pellet was assayed for total protein using BCA kit. Finally, the uptake of the micelles by caco-2 cells was evaluated in terms of supernatant drug concentration/protein mass. As a result, as shown in FIG. 2, polysaccharide 2 was most ingested with the paclitaxel suspension group, and therefore, this polysaccharide 2 was selected as the ginseng polysaccharide starting material of the present invention.
Example 8
The preparation method of the paclitaxel ginsenoside micelle modified by ginseng polysaccharide comprises the following steps:
a. weighing ginsenoside Rb 1 27mg, ginsenoside Rd 27mg, taxol 6mg in a round bottom flask, and 10mL of absolute ethanol were added and the mixture was sonicated for 30min (25 ℃ C., 40 kHz) to completely dissolve the extract.
b. The solution obtained in step a was subjected to rotary evaporation (vacuum: 100 to 120 par) at 37℃for 30min to form a uniform film.
c. B, adding 0.18mL of ginseng polysaccharide solution (with the concentration of 60 mg/mL) into the film obtained in the step b, carrying out ultrasonic treatment for 5min (25 ℃ and 40 kHz) to hydrate the film, oscillating for 0.5h at a constant temperature of 30 ℃, and filtering the obtained liquid through a 0.22um filter membrane to obtain the ginseng polysaccharide modified taxol ginsenoside micelle. The transmission electron microscope image is shown in figure 1, the particle size of the obtained micelle is 154+/-27 nm, and the PDI is 0.17+/-0.08.
Example 9
The preparation method of the paclitaxel ginsenoside micelle modified by ginseng polysaccharide comprises the following steps: step a, weighing ginsenoside Rb 1 12mg of ginsenoside Rd 12mg and taxol 6mg are placed in a round bottom flask, 0.1mL of ginseng polysaccharide solution (the concentration is 60 mg/mL) is added in step c, and the rest steps are the same as in example 8. The obtained micelle particle size was 151.+ -. 16nm and PDI was 0.15.+ -. 0.08.
Example 10
The preparation method of the paclitaxel ginsenoside micelle modified by ginseng polysaccharide comprises the following steps: weighing ginsenoside Rb 1 30mg, ginsenoside Rd30mg, taxol 6mg in a round bottom flask and the rest of the steps are the same as in example 8. The transmission electron microscope image is shown in figure 1, the particle size of the obtained micelle is 159+/-23 nm, and the PDI is 0.16+/-0.06.
Example 11
The preparation method of the docetaxel ginsenoside micelle modified by the ginseng polysaccharide comprises the following steps: weighing ginsenoside Rb 1 27mg of ginsenoside Rd 27mg and 6mg of docetaxel were placed in a round-bottomed flask, and the rest of the procedures were the same as in example 8. The particle size of the obtained micelle was 176.+ -. 33nm and the PDI was 0.11.+ -. 0.09.
Application example 1
In vitro cytotoxicity of different paclitaxel formulations on A549 cells (human non-small cell lung cancer cells)
Dilution of the cell suspension to 8X 10 4 Plates were plated at 100. Mu.L per well in 96-well plates at a volume of one/mL, and placed in a cell incubator for 24. 24 h. The cells were removed and then washed 2 times with PBS. Paclitaxel suspension, paclitaxel injection, paclitaxel ginsenoside micelle prepared in example 3, and paclitaxel ginsenoside micelle modified by panaxan prepared in example 8 are used respectivelyThe high sugar culture medium is diluted into different concentrations (0.1 mug/mL, 1 mug/mL, 5 mug/mL, 10 mug/mL, 20 mug/mL and 50 mug/mL), and the diluted paclitaxel suspension, paclitaxel injection, paclitaxel ginsenoside micelle and ginseng polysaccharide modified paclitaxel ginsenoside micelle are respectively added into each hole. Incubating in incubator for 24 h, discarding the liquid medicine and washing the cells with PBS for 2 times; adding 10 mu LCCK-8 solution (Cell Counting Kit-8 cell counting reagent) into each well, and shaking uniformly; placing the mixture into an incubator for incubation of 2h, and taking out the mixture; absorbance (a) values were measured with a microplate reader at 450 nm. As shown in FIG. 3, the ginseng polysaccharide modified paclitaxel micelle shows stronger toxicity to A549 cells at the same concentration.
Application example 2
In vitro uptake of paclitaxel ginsenoside micelle by A549 cells (human non-small cell lung cancer cells)
Dilution of the cell suspension to a concentration of 3X 10 5 Plates were plated at 24 well plates at 500 μl/well and placed in cell incubators for 24 h. The cells were removed and then washed 2 times with PBS. The paclitaxel suspension, paclitaxel injection, paclitaxel ginsenoside micelle prepared in example 3 and paclitaxel ginsenoside micelle modified by the ginseng polysaccharide prepared in example 8 are diluted to 20 mug/mL by a high sugar culture medium, and the diluted paclitaxel suspension, paclitaxel injection, paclitaxel ginsenoside micelle and paclitaxel ginsenoside micelle modified by the ginseng polysaccharide are respectively added into each hole. Incubating in incubator for 2h, discarding the liquid medicine and washing the cells with PBS for 2 times; cells were digested by adding 100. Mu.L of pancreatin, disrupted by adding 200. Mu.L of cell lysate, and centrifuged at 12000r/min for 10min. The supernatant was analyzed for drug concentration using a high performance liquid phase, and the lower protein pellet was assayed for total protein using BCA kit. Finally, the uptake of micelles by a549 cells was evaluated in terms of supernatant drug concentration/protein mass. As a result, as shown in FIG. 4, the ginseng polysaccharide-modified paclitaxel micelles were more easily taken up by A549 cells.
Application example 3
18 SD male rats were taken and randomly divided into 3 groups, weight 180-220 g. After 12h of fasting prior to dosing, different Docetaxel (DTX) formulations were administered separately: docetaxel suspension, docetaxel ginsenoside micelle prepared in example 5, and docetaxel ginsenoside micelle modified by ginseng polysaccharide prepared in example 11. The dosage is 6 mg/kg, and the blood taking time is as follows: 0.083, 0.25, 0.5, 1, 1.5, 2, 3, 4, 6, 8, 12, 24, h, blood drawn at each time point was 0.3 mL; the obtained blood is centrifuged to obtain blood plasma, and the obtained blood plasma is subjected to corresponding treatment and analysis of blood concentration, and then the absolute bioavailability is analyzed (see figure 6). The results show that the micelle group has higher bioavailability, and the bioavailability of the docetaxel ginsenoside micelle modified by the ginseng polysaccharide is highest.
Application example 4
18 SD male rats were taken and randomly divided into 3 groups, weight 180-220 g. Following a 12h pre-dosing fast, different paclitaxel formulations were administered separately: paclitaxel suspension, paclitaxel ginsenoside micelle prepared in example 3, and paclitaxel ginsenoside micelle modified by panaxan prepared in example 8. The administration dose is 50 mg/kg, and the blood taking time is as follows: 0.083, 0.25, 0.5, 1, 1.5, 2, 3, 4, 6, 8, 12, 24, h, blood drawn at each time point was 0.3 mL; the obtained blood is centrifuged to obtain blood plasma, and the blood plasma is subjected to corresponding treatment and analysis of blood concentration, and bioavailability of each preparation is compared (see figure 5). The results show that the micelle group has higher bioavailability, and the bioavailability of the ginseng polysaccharide modified taxol-ginsenoside micelle is highest.
Application example 5
Research on anti-tumor effect of micelle on nude mice
A549 cells were cultured in DMEM medium containing 10% fetal bovine serum. A549 cells in exponential growth phase were collected and resuspended in serum-free medium to appropriate concentrations (2.5X10 6 ) After that, it was used for subcutaneous tumor inoculation of mice. Subcutaneous inoculation of 6 female mice on the right side 2.5X10 6 A549 cells. Average volume of tumor is 200-300mm 3 At the time, A549 (generation 3) tumor blocks which are inoculated to the armpit of a nude mouse and are in a rapid proliferation stage are taken, the tumor blocks are cut into tumor blocks with the size of 1mm multiplied by 1mm, and the nude mouse is inoculated subcutaneously on the right limb of the nude mouse by using a trocar under the aseptic condition. Until the tumor grows to 300-400mm 3 Time-division group. Randomly grouped according to the size of the tumor,marked as the following 6 groups respectively: physiological saline group, paclitaxel suspension group (10 mg/kg), paclitaxel ginsenoside micelle group prepared in example 3 (10 mg/kg), panaxan modified paclitaxel ginsenoside micelle low dose group prepared in example 8 (5 mg/kg), panaxan modified paclitaxel ginsenoside micelle medium dose group (10 mg/kg), panaxan modified paclitaxel ginsenoside micelle high dose group (20 mg/kg). Eight weeks after grouping. After tumor inoculation, routine monitoring mainly includes tumor growth and treatment of the animal weight gain or loss (weight 2 times per week).
Relative tumor inhibition rate TGI (%): tgi=1-T/C (%). T/C% is the relative tumor proliferation rate, i.e., the percentage value of the treated and control groups relative to the tumor volume or tumor weight at a certain time point. T and C are the Relative Tumor Volume (RTV) or Tumor Weight (TW) of the treatment group and the control group, respectively, at a particular time point.
From the data of the relative tumor inhibition TGI (%) (tgi= (1-treatment group tumor weight/control group tumor weight) ×100%) of each drug group (see table 7), the effect of each drug group on tumor was found to be: the high-dose group of the paclitaxel micelle modified by the ginseng polysaccharide is more than the paclitaxel ginsenoside micelle group, the paclitaxel suspension group is more than the low-dose group of the paclitaxel micelle modified by the ginseng polysaccharide, and the result shows that the paclitaxel micelle of the ginseng polysaccharide has good inhibition effect on tumors.
TABLE 7 therapeutic Effect of different formulations on tumors
Application example 6
Initial safety evaluation of ginseng polysaccharide modified paclitaxel micelle
50 SPF-grade Kunming mice (body weight 20.+ -. 2 g /) were kept in an environment with regular light and dark alternation time, and different paclitaxel preparations were administered separately: blank group, paclitaxel suspension group (10 mg/mL), ginsenoside group, paclitaxel injection group (10 mg/mL), paclitaxel ginsenoside micelle low dose group (5 mg/mL) prepared in example 3, paclitaxel ginsenoside micelle medium dose group (10 mg/mL), paclitaxel ginsenoside micelle high dose group (20 mg/mL), ginseng polysaccharide modified paclitaxel ginsenoside micelle low dose group (5 mg/mL) prepared in example 8, ginseng polysaccharide modified paclitaxel ginsenoside micelle medium dose group (10 mg/mL), and ginseng polysaccharide modified paclitaxel ginsenoside micelle high dose group (20 mg/mL). The administration is carried out daily for 10 days, abnormal actions such as torsion, convulsion and the like are observed after the administration, and the weight, activity, feeding and drinking conditions, urination and defecation conditions, unhairing conditions and the like of mice in each group are observed and recorded daily. The safety was primarily evaluated by examining the change in the latency of the pain response during administration and blood routine related indicators, taking the occurrence of dose-limiting toxicity (neuropathic pain and neutropenia) as an evaluation target for safety. Mice were assayed for thermal pain response latency (TWL) using a hotplate method. The hotplate was adjusted to 55±0.5 ℃ with the time between full landing of the mice on their hind feet to licking the hind feet or jumping as their thermal pain response latency (TWL). Mice with pain response latency within 5-20 s were screened for experiments prior to the experiments. To avoid injury to the soles of the mice, the mice were returned to their cages with a score of 40 s that remained stationary beyond the maximum response time of 40 s. The hot plate measurement is carried out the day before the administration to obtain the stable basic reaction time before the experiment, and the measurement of the hot plate is measured before the administration during the administration so as to avoid inaccurate measurement results caused by the influence of the emotion of the mice in the administration process. Three consecutive determinations were made at intervals of 10min, and the average value was taken as TWL of the mice on the same day. EP tubes of 1.5. 1.5 mL were prepared in advance, and anticoagulant 0.15. 0.15 mL was added to each tube. Mice were harvested from eyeballs and placed in EP tubes on day 10 post-dose, immediately sampled for detection by a blood-plasma analyzer, focusing on changes in the numbers of leukocytes, neutrophils, platelets, hemoglobin.
The results were counted: the examination results of neuropathic pain (see figure 7) show that compared with paclitaxel injection, paclitaxel ginsenoside micelle and paclitaxel ginsenoside micelle modified by ginseng polysaccharide have the advantages of delayed time for inducing neuropathic pain, maximum delay of paclitaxel ginsenoside micelle modified by ginseng polysaccharide and better safety. The results of hematology toxicity examination (see figure 8) show that the paclitaxel ginsenoside micelle modified by the ginseng polysaccharide has little difference in the indexes of platelets, hemoglobin, white blood cells and neutrophils compared with the paclitaxel injection under the same administration dosage.
Claims (10)
1. An oral micelle taking ginseng active ingredients as auxiliary materials is characterized in that: the oral micelle comprises auxiliary materials and an anti-tumor indissoluble drug, wherein the auxiliary materials are ginsenoside, the mass ratio of the anti-tumor indissoluble drug to the ginsenoside is 1 (0.2-30), and the anti-tumor indissoluble drug is paclitaxel or docetaxel.
2. The oral micelle taking ginseng active ingredients as auxiliary materials according to claim 1, wherein: the ginsenoside is one or a combination of more of ginsenoside Ro, ri, rh3, F4, F11, RT5, RT4, rb1, rb2, rb3, rc, rd, rh2, re, rg1, rg2, rf, PPD and Rh 1.
3. The oral micelle taking ginseng active ingredients as auxiliary materials according to claim 1, wherein: the oral micelle also comprises a modified material ginseng polysaccharide, wherein the mass ratio of the ginseng polysaccharide to the ginsenoside is 1 (0.05-10).
4. An oral micelle containing ginseng active ingredients as an auxiliary material according to claim 1 or 3, wherein: the mass ratio of the anti-tumor indissoluble medicine to the ginsenoside is 1 (0.2-9), and the mass ratio of the ginseng polysaccharide to the ginsenoside is 1 (0.2-5).
5. An oral micelle containing ginseng active ingredients as set forth in claim 3, wherein: the extraction method of ginseng polysaccharide comprises the following steps:
a. adding pure water into ginseng sample according to a feed liquid ratio of 1:30, extracting for 0.5-1h, cooling, centrifuging, precipitating with ethanol, filtering, collecting precipitate, and drying to obtain ginseng crude polysaccharide;
b. and d, performing enzymolysis by adopting alpha-amylase to remove starch in the ginseng crude polysaccharide obtained in the step a, wherein the enzymolysis conditions are as follows: 800 U alpha-amylase, and carrying out enzymolysis for 0.5-1h at 50-60 ℃;
c. and d, carrying out enzymolysis by papain to remove protein in the ginseng crude polysaccharide obtained in the step b, wherein the enzymolysis conditions are as follows: 200 U papain is subjected to enzymolysis at 30-40 ℃ for 1-3 h to obtain crude ginseng polysaccharide after enzymolysis;
d. preparing crude ginseng polysaccharide after enzymolysis, performing column chromatography elution by using DEAE-52 cellulose filler, eluting with distilled water, 0.1, 0.2, 0.3, 0.4, 0.5, 0.6, 0.7 and 0.8mol/L NaCl solution respectively in sequence, eluting at the flow rate of 1mL/min, collecting eluent to obtain polysaccharide 1, 2, 3, 4, 5, 6, 7, 8 and 9 respectively, and obtaining polysaccharide 2 to obtain the ginseng polysaccharide.
6. A method for preparing oral micelles with ginseng active ingredients as in any one of claims 1-3 or 5, comprising the steps of:
(1) Mixing ginsenoside and antineoplastic insoluble medicine, placing in absolute ethanol, and dissolving by ultrasound to obtain medicine and ginsenoside-containing solution;
(2) The solution containing the medicine and the ginsenoside obtained in the step (1) is subjected to rotary evaporation at the temperature of 30-60 ℃ to remove the ethanol, so that a uniform film is formed;
(3) Adding purified water or ginseng polysaccharide water solution into the film obtained in the step (2), carrying out ultrasonic treatment at the temperature of 20-55 ℃ for 0.5-15 min to hydrate the film, carrying out constant-temperature vibration at the temperature of 30-40 ℃ for 0.5-2 h, and then filtering with a filter membrane with the thickness of 0.22 mu m to obtain the oral micelle.
7. The method of manufacturing according to claim 6, wherein: in the step (1), the ultrasonic time is 10-40 min, and the concentration of the anti-tumor indissoluble medicine in the medicine-containing and ginsenoside solution is 0.01-20 mg/mL; in the step (2), the time of rotary evaporation is 5-60 min.
8. The method of manufacturing according to claim 7, wherein: the concentration of the anti-tumor indissoluble medicine in the step (1) is 0.1-0.6 mg/mL.
9. Use of an oral micelle taking ginseng active ingredients as auxiliary materials according to any one of claims 1 to 3 or 5 in preparing an anti-tumor medicament.
10. Use of an oral micelle containing ginseng as an active ingredient as claimed in claim 2 for improving oral absorption of paclitaxel or docetaxel.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202311819562.4A CN117752607A (en) | 2023-12-27 | 2023-12-27 | Oral micelle taking ginseng active ingredient as auxiliary material and preparation method and application thereof |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202311819562.4A CN117752607A (en) | 2023-12-27 | 2023-12-27 | Oral micelle taking ginseng active ingredient as auxiliary material and preparation method and application thereof |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN117752607A true CN117752607A (en) | 2024-03-26 |
Family
ID=90325433
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN202311819562.4A Pending CN117752607A (en) | 2023-12-27 | 2023-12-27 | Oral micelle taking ginseng active ingredient as auxiliary material and preparation method and application thereof |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN117752607A (en) |
-
2023
- 2023-12-27 CN CN202311819562.4A patent/CN117752607A/en active Pending
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| US20110059124A1 (en) | The quality control method and application of a kind of ganoderma lucidum spore oil fat emulsion | |
| WO2015090180A1 (en) | Sanchi flower arab galactan and preparation method and use thereof | |
| CN104644697B (en) | The preparation method and applications of ganoderma lucidum Ultramicro-powder | |
| EP3572088B1 (en) | A panax plant extract and pharmaceutical composition and use thereof | |
| Ke et al. | Research progress on the extraction technology and activity study of Epimedium polysaccharides | |
| CN101301455A (en) | Chinese medicine compound turmeric rhizome solid dispersion for treating hyperlipemia | |
| CN105338993A (en) | Composition for preventing, relieving or treating colitis, containing complex extracts | |
| CN108186559A (en) | Cystine injection and its preparation process | |
| CN109674742A (en) | A kind of curcumin hydrogel matrix slow releasing composition, preparation method and its application in anti-cancer field | |
| US20050287230A1 (en) | Method of producing ginsenoside 20 (R)-Rh2 and composition of matter thereof | |
| CN1977885B (en) | Antihepatitis medicinal composition | |
| CN102357180B (en) | Traditional Chinese medicine composite for curing cancer and preparation method and application thereof | |
| CN101428050B (en) | Active composition for treating thrombus, cardio-cerebrovascular system diseases | |
| CN1919339B (en) | Cucurbitacin nano preparation comprising protein, preparation method and use thereof | |
| CN117752607A (en) | Oral micelle taking ginseng active ingredient as auxiliary material and preparation method and application thereof | |
| WO2020000828A1 (en) | Gracilaria lemaneiformis polysaccharide having significant hypolipidemic activity and preparation method therefor and use thereof | |
| CN101152233B (en) | Pharmaceutical composition of snakegourd fruit and folium ginkgo | |
| CN107266599B (en) | Flammulina velutipes, extracting method and its application in terms of functional consitipation drug is treated in preparation | |
| CN106309758B (en) | Pharmaceutical composition for resisting gastrointestinal cancer | |
| CN100482266C (en) | Medical composite prepared by sarcandra and oldenlandia | |
| CN103330721A (en) | Holothurian flower extract for treatment of female POF and its preparation method | |
| CN100534476C (en) | Traditional Chinese medicine composition made by isatis root and rhizoma belamcandae, and its preparation method and use | |
| CN111329871A (en) | Preparation method and application of product of cordyceps militaris for preventing and treating liver cancer | |
| CN101011543A (en) | Antineoplastic medicine composition | |
| CN110721193B (en) | Application of cynomorium songaricum total polysaccharide in preparation of medicine for treating asthma |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination |