CN104056275B - 多功能主动靶向透明质酸‑聚乳酸载体合成及其抗肿瘤药物胶束制备方法 - Google Patents
多功能主动靶向透明质酸‑聚乳酸载体合成及其抗肿瘤药物胶束制备方法 Download PDFInfo
- Publication number
- CN104056275B CN104056275B CN201410244165.3A CN201410244165A CN104056275B CN 104056275 B CN104056275 B CN 104056275B CN 201410244165 A CN201410244165 A CN 201410244165A CN 104056275 B CN104056275 B CN 104056275B
- Authority
- CN
- China
- Prior art keywords
- pla
- micelle
- hyaluronic acid
- antineoplastic
- suspension
- 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.)
- Expired - Fee Related
Links
- 229920002674 hyaluronan Polymers 0.000 title claims abstract description 41
- 229960003160 hyaluronic acid Drugs 0.000 title claims abstract description 37
- KIUKXJAPPMFGSW-DNGZLQJQSA-N (2S,3S,4S,5R,6R)-6-[(2S,3R,4R,5S,6R)-3-Acetamido-2-[(2S,3S,4R,5R,6R)-6-[(2R,3R,4R,5S,6R)-3-acetamido-2,5-dihydroxy-6-(hydroxymethyl)oxan-4-yl]oxy-2-carboxy-4,5-dihydroxyoxan-3-yl]oxy-5-hydroxy-6-(hydroxymethyl)oxan-4-yl]oxy-3,4,5-trihydroxyoxane-2-carboxylic acid Chemical compound CC(=O)N[C@H]1[C@H](O)O[C@H](CO)[C@@H](O)[C@@H]1O[C@H]1[C@H](O)[C@@H](O)[C@H](O[C@H]2[C@@H]([C@@H](O[C@H]3[C@@H]([C@@H](O)[C@H](O)[C@H](O3)C(O)=O)O)[C@H](O)[C@@H](CO)O2)NC(C)=O)[C@@H](C(O)=O)O1 KIUKXJAPPMFGSW-DNGZLQJQSA-N 0.000 title claims abstract description 35
- 230000000118 anti-neoplastic effect Effects 0.000 title claims abstract description 24
- 230000008685 targeting Effects 0.000 title claims abstract description 22
- 238000002360 preparation method Methods 0.000 title claims abstract description 19
- 238000003786 synthesis reaction Methods 0.000 title claims abstract description 18
- 230000015572 biosynthetic process Effects 0.000 title claims abstract description 17
- 239000000693 micelle Substances 0.000 claims abstract description 92
- 239000004626 polylactic acid Substances 0.000 claims abstract description 87
- 229920000747 poly(lactic acid) Polymers 0.000 claims abstract description 74
- 239000003814 drug Substances 0.000 claims abstract description 38
- 239000002131 composite material Substances 0.000 claims abstract description 37
- 239000000203 mixture Substances 0.000 claims abstract description 32
- 229920000642 polymer Polymers 0.000 claims abstract description 27
- 238000001338 self-assembly Methods 0.000 claims abstract description 25
- 229920000728 polyester Polymers 0.000 claims abstract description 21
- 239000000843 powder Substances 0.000 claims abstract description 11
- 239000003795 chemical substances by application Substances 0.000 claims abstract description 6
- 229920001577 copolymer Polymers 0.000 claims abstract description 4
- 210000003734 kidney Anatomy 0.000 claims abstract description 4
- 239000000725 suspension Substances 0.000 claims description 39
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Chemical compound O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 31
- 229960001592 paclitaxel Drugs 0.000 claims description 28
- 229930012538 Paclitaxel Natural products 0.000 claims description 27
- IAZDPXIOMUYVGZ-UHFFFAOYSA-N Dimethylsulphoxide Chemical compound CS(C)=O IAZDPXIOMUYVGZ-UHFFFAOYSA-N 0.000 claims description 26
- 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 description 26
- 239000008213 purified water Substances 0.000 claims description 23
- AOJJSUZBOXZQNB-TZSSRYMLSA-N Doxorubicin Chemical compound O([C@H]1C[C@@](O)(CC=2C(O)=C3C(=O)C=4C=CC=C(C=4C(=O)C3=C(O)C=21)OC)C(=O)CO)[C@H]1C[C@H](N)[C@H](O)[C@H](C)O1 AOJJSUZBOXZQNB-TZSSRYMLSA-N 0.000 claims description 22
- 239000003960 organic solvent Substances 0.000 claims description 22
- WEVYAHXRMPXWCK-UHFFFAOYSA-N Acetonitrile Chemical compound CC#N WEVYAHXRMPXWCK-UHFFFAOYSA-N 0.000 claims description 21
- 238000004108 freeze drying Methods 0.000 claims description 21
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 claims description 19
- CSCPPACGZOOCGX-UHFFFAOYSA-N Acetone Chemical compound CC(C)=O CSCPPACGZOOCGX-UHFFFAOYSA-N 0.000 claims description 18
- RTZKZFJDLAIYFH-UHFFFAOYSA-N Diethyl ether Chemical compound CCOCC RTZKZFJDLAIYFH-UHFFFAOYSA-N 0.000 claims description 18
- OKKJLVBELUTLKV-UHFFFAOYSA-N Methanol Chemical compound OC OKKJLVBELUTLKV-UHFFFAOYSA-N 0.000 claims description 18
- YMWUJEATGCHHMB-UHFFFAOYSA-N Dichloromethane Chemical compound ClCCl YMWUJEATGCHHMB-UHFFFAOYSA-N 0.000 claims description 15
- 229920001400 block copolymer Polymers 0.000 claims description 15
- 229940079593 drug Drugs 0.000 claims description 15
- 239000007787 solid Substances 0.000 claims description 14
- IJGRMHOSHXDMSA-UHFFFAOYSA-N Atomic nitrogen Chemical compound N#N IJGRMHOSHXDMSA-UHFFFAOYSA-N 0.000 claims description 12
- HEDRZPFGACZZDS-UHFFFAOYSA-N Chloroform Chemical compound ClC(Cl)Cl HEDRZPFGACZZDS-UHFFFAOYSA-N 0.000 claims description 12
- 238000013019 agitation Methods 0.000 claims description 12
- 239000007788 liquid Substances 0.000 claims description 12
- 239000000523 sample Substances 0.000 claims description 11
- XEKOWRVHYACXOJ-UHFFFAOYSA-N Ethyl acetate Chemical compound CCOC(C)=O XEKOWRVHYACXOJ-UHFFFAOYSA-N 0.000 claims description 9
- 238000001914 filtration Methods 0.000 claims description 9
- VEXZGXHMUGYJMC-UHFFFAOYSA-N Hydrochloric acid Chemical compound Cl VEXZGXHMUGYJMC-UHFFFAOYSA-N 0.000 claims description 8
- 238000004090 dissolution Methods 0.000 claims description 8
- 239000012982 microporous membrane Substances 0.000 claims description 8
- 239000012071 phase Substances 0.000 claims description 8
- 239000002904 solvent Substances 0.000 claims description 8
- 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 description 7
- -1 acyls Imines Chemical class 0.000 claims description 7
- 229940009456 adriamycin Drugs 0.000 claims description 7
- 239000008346 aqueous phase Substances 0.000 claims description 7
- 238000006243 chemical reaction Methods 0.000 claims description 7
- 238000000502 dialysis Methods 0.000 claims description 7
- 238000009792 diffusion process Methods 0.000 claims description 7
- 229960003668 docetaxel Drugs 0.000 claims description 7
- 238000003760 magnetic stirring Methods 0.000 claims description 6
- 229910052757 nitrogen Inorganic materials 0.000 claims description 6
- 229920001610 polycaprolactone Polymers 0.000 claims description 6
- 238000005516 engineering process Methods 0.000 claims description 5
- 238000001291 vacuum drying Methods 0.000 claims description 5
- JGSARLDLIJGVTE-UHFFFAOYSA-N 3,3-dimethyl-7-oxo-6-[(2-phenylacetyl)amino]-4-thia-1-azabicyclo[3.2.0]heptane-2-carboxylic acid Chemical compound O=C1N2C(C(O)=O)C(C)(C)SC2C1NC(=O)CC1=CC=CC=C1 JGSARLDLIJGVTE-UHFFFAOYSA-N 0.000 claims description 4
- QOSSAOTZNIDXMA-UHFFFAOYSA-N Dicylcohexylcarbodiimide Chemical compound C1CCCCC1N=C=NC1CCCCC1 QOSSAOTZNIDXMA-UHFFFAOYSA-N 0.000 claims description 4
- FBOZXECLQNJBKD-ZDUSSCGKSA-N L-methotrexate Chemical compound C=1N=C2N=C(N)N=C(N)C2=NC=1CN(C)C1=CC=C(C(=O)N[C@@H](CCC(O)=O)C(O)=O)C=C1 FBOZXECLQNJBKD-ZDUSSCGKSA-N 0.000 claims description 4
- 239000002253 acid Substances 0.000 claims description 4
- 230000004913 activation Effects 0.000 claims description 4
- 239000003153 chemical reaction reagent Substances 0.000 claims description 4
- VFLDPWHFBUODDF-FCXRPNKRSA-N curcumin Chemical compound C1=C(O)C(OC)=CC(\C=C\C(=O)CC(=O)\C=C\C=2C=C(OC)C(O)=CC=2)=C1 VFLDPWHFBUODDF-FCXRPNKRSA-N 0.000 claims description 4
- 229960000485 methotrexate Drugs 0.000 claims description 4
- 239000012046 mixed solvent Substances 0.000 claims description 4
- 238000012856 packing Methods 0.000 claims description 4
- 238000001556 precipitation Methods 0.000 claims description 4
- 238000000967 suction filtration Methods 0.000 claims description 4
- 229920000428 triblock copolymer Polymers 0.000 claims description 4
- HDTRYLNUVZCQOY-UHFFFAOYSA-N α-D-glucopyranosyl-α-D-glucopyranoside Natural products OC1C(O)C(O)C(CO)OC1OC1C(O)C(O)C(O)C(CO)O1 HDTRYLNUVZCQOY-UHFFFAOYSA-N 0.000 claims description 3
- HAWSQZCWOQZXHI-FQEVSTJZSA-N 10-Hydroxycamptothecin Chemical compound C1=C(O)C=C2C=C(CN3C4=CC5=C(C3=O)COC(=O)[C@]5(O)CC)C4=NC2=C1 HAWSQZCWOQZXHI-FQEVSTJZSA-N 0.000 claims description 3
- STQGQHZAVUOBTE-UHFFFAOYSA-N 7-Cyan-hept-2t-en-4,6-diinsaeure Natural products C1=2C(O)=C3C(=O)C=4C(OC)=CC=CC=4C(=O)C3=C(O)C=2CC(O)(C(C)=O)CC1OC1CC(N)C(O)C(C)O1 STQGQHZAVUOBTE-UHFFFAOYSA-N 0.000 claims description 3
- FBPFZTCFMRRESA-KVTDHHQDSA-N D-Mannitol Chemical compound OC[C@@H](O)[C@@H](O)[C@H](O)[C@H](O)CO FBPFZTCFMRRESA-KVTDHHQDSA-N 0.000 claims description 3
- GUBGYTABKSRVRQ-QKKXKWKRSA-N Lactose Natural products OC[C@H]1O[C@@H](O[C@H]2[C@H](O)[C@@H](O)C(O)O[C@@H]2CO)[C@H](O)[C@@H](O)[C@H]1O GUBGYTABKSRVRQ-QKKXKWKRSA-N 0.000 claims description 3
- 229930195725 Mannitol Natural products 0.000 claims description 3
- 208000005718 Stomach Neoplasms Diseases 0.000 claims description 3
- HDTRYLNUVZCQOY-WSWWMNSNSA-N Trehalose Natural products O[C@@H]1[C@@H](O)[C@@H](O)[C@@H](CO)O[C@@H]1O[C@@H]1[C@H](O)[C@@H](O)[C@@H](O)[C@@H](CO)O1 HDTRYLNUVZCQOY-WSWWMNSNSA-N 0.000 claims description 3
- 208000006105 Uterine Cervical Neoplasms Diseases 0.000 claims description 3
- HDTRYLNUVZCQOY-LIZSDCNHSA-N alpha,alpha-trehalose Chemical compound O[C@@H]1[C@@H](O)[C@H](O)[C@@H](CO)O[C@@H]1O[C@@H]1[C@H](O)[C@@H](O)[C@H](O)[C@@H](CO)O1 HDTRYLNUVZCQOY-LIZSDCNHSA-N 0.000 claims description 3
- 150000001412 amines Chemical group 0.000 claims description 3
- 201000010881 cervical cancer Diseases 0.000 claims description 3
- 238000005253 cladding Methods 0.000 claims description 3
- 229960000975 daunorubicin Drugs 0.000 claims description 3
- STQGQHZAVUOBTE-VGBVRHCVSA-N daunorubicin Chemical compound O([C@H]1C[C@@](O)(CC=2C(O)=C3C(=O)C=4C=CC=C(C=4C(=O)C3=C(O)C=21)OC)C(C)=O)[C@H]1C[C@H](N)[C@H](O)[C@H](C)O1 STQGQHZAVUOBTE-VGBVRHCVSA-N 0.000 claims description 3
- 206010017758 gastric cancer Diseases 0.000 claims description 3
- 229920000578 graft copolymer Polymers 0.000 claims description 3
- KIUKXJAPPMFGSW-MNSSHETKSA-N hyaluronan Chemical compound CC(=O)N[C@H]1[C@H](O)O[C@H](CO)[C@@H](O)C1O[C@H]1[C@H](O)[C@@H](O)[C@H](O[C@H]2[C@@H](C(O[C@H]3[C@@H]([C@@H](O)[C@H](O)[C@H](O3)C(O)=O)O)[C@H](O)[C@@H](CO)O2)NC(C)=O)[C@@H](C(O)=O)O1 KIUKXJAPPMFGSW-MNSSHETKSA-N 0.000 claims description 3
- 229940099552 hyaluronan Drugs 0.000 claims description 3
- 230000002209 hydrophobic effect Effects 0.000 claims description 3
- JJTUDXZGHPGLLC-UHFFFAOYSA-N lactide Chemical compound CC1OC(=O)C(C)OC1=O JJTUDXZGHPGLLC-UHFFFAOYSA-N 0.000 claims description 3
- 239000008101 lactose Substances 0.000 claims description 3
- 208000032839 leukemia Diseases 0.000 claims description 3
- 201000007270 liver cancer Diseases 0.000 claims description 3
- 208000014018 liver neoplasm Diseases 0.000 claims description 3
- 239000000594 mannitol Substances 0.000 claims description 3
- 235000010355 mannitol Nutrition 0.000 claims description 3
- 238000002156 mixing Methods 0.000 claims description 3
- UDGSVBYJWHOHNN-UHFFFAOYSA-N n',n'-diethylethane-1,2-diamine Chemical class CCN(CC)CCN UDGSVBYJWHOHNN-UHFFFAOYSA-N 0.000 claims description 3
- 239000012074 organic phase Substances 0.000 claims description 3
- 239000008363 phosphate buffer Substances 0.000 claims description 3
- 239000000376 reactant Substances 0.000 claims description 3
- 201000011549 stomach cancer Diseases 0.000 claims description 3
- 238000010792 warming Methods 0.000 claims description 3
- 108010082627 (ethylene glycol)-poly(lactide)-poly(3-methyl-morpholine-2,5-dione) Proteins 0.000 claims description 2
- AOJJSUZBOXZQNB-VTZDEGQISA-N 4'-epidoxorubicin Chemical compound O([C@H]1C[C@@](O)(CC=2C(O)=C3C(=O)C=4C=CC=C(C=4C(=O)C3=C(O)C=21)OC)C(=O)CO)[C@H]1C[C@H](N)[C@@H](O)[C@H](C)O1 AOJJSUZBOXZQNB-VTZDEGQISA-N 0.000 claims description 2
- GUBGYTABKSRVRQ-XLOQQCSPSA-N Alpha-Lactose Chemical compound O[C@@H]1[C@@H](O)[C@@H](O)[C@@H](CO)O[C@H]1O[C@@H]1[C@@H](CO)O[C@H](O)[C@H](O)[C@H]1O GUBGYTABKSRVRQ-XLOQQCSPSA-N 0.000 claims description 2
- 206010006187 Breast cancer Diseases 0.000 claims description 2
- 208000026310 Breast neoplasm Diseases 0.000 claims description 2
- HTIJFSOGRVMCQR-UHFFFAOYSA-N Epirubicin Natural products COc1cccc2C(=O)c3c(O)c4CC(O)(CC(OC5CC(N)C(=O)C(C)O5)c4c(O)c3C(=O)c12)C(=O)CO HTIJFSOGRVMCQR-UHFFFAOYSA-N 0.000 claims description 2
- WQZGKKKJIJFFOK-GASJEMHNSA-N Glucose Natural products OC[C@H]1OC(O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-GASJEMHNSA-N 0.000 claims description 2
- 235000005035 Panax pseudoginseng ssp. pseudoginseng Nutrition 0.000 claims description 2
- 235000003140 Panax quinquefolius Nutrition 0.000 claims description 2
- 229920000436 Poly(lactide-co-glycolide)-block-poly(ethylene glycol)-block-poly(lactide-co-glycolide) Polymers 0.000 claims description 2
- 229920000432 Polylactide-block-poly(ethylene glycol)-block-polylactide Polymers 0.000 claims description 2
- 229930006000 Sucrose Natural products 0.000 claims description 2
- CZMRCDWAGMRECN-UGDNZRGBSA-N Sucrose Chemical compound O[C@H]1[C@H](O)[C@@H](CO)O[C@@]1(CO)O[C@@H]1[C@H](O)[C@@H](O)[C@H](O)[C@@H](CO)O1 CZMRCDWAGMRECN-UGDNZRGBSA-N 0.000 claims description 2
- 125000003368 amide group Chemical group 0.000 claims description 2
- 238000005576 amination reaction Methods 0.000 claims description 2
- WQZGKKKJIJFFOK-VFUOTHLCSA-N beta-D-glucose Chemical compound OC[C@H]1O[C@@H](O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-VFUOTHLCSA-N 0.000 claims description 2
- 125000003178 carboxy group Chemical group [H]OC(*)=O 0.000 claims description 2
- 238000011109 contamination Methods 0.000 claims description 2
- 229940109262 curcumin Drugs 0.000 claims description 2
- 235000012754 curcumin Nutrition 0.000 claims description 2
- 239000004148 curcumin Substances 0.000 claims description 2
- VFLDPWHFBUODDF-UHFFFAOYSA-N diferuloylmethane Natural products C1=C(O)C(OC)=CC(C=CC(=O)CC(=O)C=CC=2C=C(OC)C(O)=CC=2)=C1 VFLDPWHFBUODDF-UHFFFAOYSA-N 0.000 claims description 2
- 230000008030 elimination Effects 0.000 claims description 2
- 238000003379 elimination reaction Methods 0.000 claims description 2
- 229960001904 epirubicin Drugs 0.000 claims description 2
- 235000008434 ginseng Nutrition 0.000 claims description 2
- 239000008103 glucose Substances 0.000 claims description 2
- 125000002887 hydroxy group Chemical group [H]O* 0.000 claims description 2
- KKZJGLLVHKMTCM-UHFFFAOYSA-N mitoxantrone Chemical compound O=C1C2=C(O)C=CC(O)=C2C(=O)C2=C1C(NCCNCCO)=CC=C2NCCNCCO KKZJGLLVHKMTCM-UHFFFAOYSA-N 0.000 claims description 2
- 229960001156 mitoxantrone Drugs 0.000 claims description 2
- 208000002154 non-small cell lung carcinoma Diseases 0.000 claims description 2
- SDHTXBWLVGWJFT-XKCURVIJSA-N oridonin Chemical compound C([C@@H]1[C@@H](O)[C@@]23C(C1=C)=O)C[C@H]2[C@]12[C@@H](O)CCC(C)(C)[C@H]1[C@H](O)[C@@]3(O)OC2 SDHTXBWLVGWJFT-XKCURVIJSA-N 0.000 claims description 2
- 239000003208 petroleum Substances 0.000 claims description 2
- 229920000762 poly(caprolactone)-poly(ethylene glycol)-poly(caprolactone) Polymers 0.000 claims description 2
- 229920000382 poly(ethylene glycol) methyl ether-block-poly(L-lactide-co-glycolide) Polymers 0.000 claims description 2
- 108010054442 polyalanine Proteins 0.000 claims description 2
- 108010094020 polyglycine Proteins 0.000 claims description 2
- 229920000232 polyglycine polymer Polymers 0.000 claims description 2
- 239000001397 quillaja saponaria molina bark Substances 0.000 claims description 2
- 229930185378 rubescensin Natural products 0.000 claims description 2
- VHXNKPBCCMUMSW-FQEVSTJZSA-N rubitecan Chemical compound C1=CC([N+]([O-])=O)=C2C=C(CN3C4=CC5=C(C3=O)COC(=O)[C@]5(O)CC)C4=NC2=C1 VHXNKPBCCMUMSW-FQEVSTJZSA-N 0.000 claims description 2
- 229950009213 rubitecan Drugs 0.000 claims description 2
- 229930182490 saponin Natural products 0.000 claims description 2
- 150000007949 saponins Chemical class 0.000 claims description 2
- 229910000033 sodium borohydride Inorganic materials 0.000 claims description 2
- 239000012279 sodium borohydride Substances 0.000 claims description 2
- BEOOHQFXGBMRKU-UHFFFAOYSA-N sodium cyanoborohydride Chemical group [Na+].[B-]C#N BEOOHQFXGBMRKU-UHFFFAOYSA-N 0.000 claims description 2
- 239000005720 sucrose Substances 0.000 claims description 2
- 208000029729 tumor suppressor gene on chromosome 11 Diseases 0.000 claims description 2
- QGZKDVFQNNGYKY-UHFFFAOYSA-N Ammonia Chemical compound N QGZKDVFQNNGYKY-UHFFFAOYSA-N 0.000 claims 2
- JVTAAEKCZFNVCJ-UHFFFAOYSA-N lactic acid Chemical compound CC(O)C(O)=O JVTAAEKCZFNVCJ-UHFFFAOYSA-N 0.000 claims 2
- NAALWFYYHHJEFQ-ZASNTINBSA-N (2s,5r,6r)-6-[[(2r)-2-[[6-[4-[bis(2-hydroxyethyl)sulfamoyl]phenyl]-2-oxo-1h-pyridine-3-carbonyl]amino]-2-(4-hydroxyphenyl)acetyl]amino]-3,3-dimethyl-7-oxo-4-thia-1-azabicyclo[3.2.0]heptane-2-carboxylic acid Chemical compound N([C@@H](C(=O)N[C@H]1[C@H]2SC([C@@H](N2C1=O)C(O)=O)(C)C)C=1C=CC(O)=CC=1)C(=O)C(C(N1)=O)=CC=C1C1=CC=C(S(=O)(=O)N(CCO)CCO)C=C1 NAALWFYYHHJEFQ-ZASNTINBSA-N 0.000 claims 1
- 241000208340 Araliaceae Species 0.000 claims 1
- 108010020346 Polyglutamic Acid Proteins 0.000 claims 1
- 238000012644 addition polymerization Methods 0.000 claims 1
- 229910021529 ammonia Inorganic materials 0.000 claims 1
- 238000006555 catalytic reaction Methods 0.000 claims 1
- 229960003109 daunorubicin hydrochloride Drugs 0.000 claims 1
- 150000002148 esters Chemical class 0.000 claims 1
- VJJPUSNTGOMMGY-MRVIYFEKSA-N etoposide Chemical compound COC1=C(O)C(OC)=CC([C@@H]2C3=CC=4OCOC=4C=C3[C@@H](O[C@H]3[C@@H]([C@@H](O)[C@@H]4O[C@H](C)OC[C@H]4O3)O)[C@@H]3[C@@H]2C(OC3)=O)=C1 VJJPUSNTGOMMGY-MRVIYFEKSA-N 0.000 claims 1
- 229960005420 etoposide Drugs 0.000 claims 1
- 238000007710 freezing Methods 0.000 claims 1
- 230000008014 freezing Effects 0.000 claims 1
- 239000012535 impurity Substances 0.000 claims 1
- 239000004310 lactic acid Substances 0.000 claims 1
- 235000014655 lactic acid Nutrition 0.000 claims 1
- 229920002643 polyglutamic acid Polymers 0.000 claims 1
- 230000010148 water-pollination Effects 0.000 claims 1
- 206010028980 Neoplasm Diseases 0.000 abstract description 33
- 238000000034 method Methods 0.000 abstract description 19
- 230000000694 effects Effects 0.000 abstract description 10
- 238000002347 injection Methods 0.000 abstract description 9
- 239000007924 injection Substances 0.000 abstract description 9
- 210000000865 mononuclear phagocyte system Anatomy 0.000 abstract description 8
- 230000003285 pharmacodynamic effect Effects 0.000 abstract description 7
- 230000009471 action Effects 0.000 abstract description 5
- 238000009472 formulation Methods 0.000 abstract description 4
- 206010057249 Phagocytosis Diseases 0.000 abstract description 3
- 238000001727 in vivo Methods 0.000 abstract description 3
- 230000008782 phagocytosis Effects 0.000 abstract description 3
- 230000029142 excretion Effects 0.000 abstract description 2
- 239000003446 ligand Substances 0.000 abstract description 2
- 239000000825 pharmaceutical preparation Substances 0.000 abstract description 2
- 230000001093 anti-cancer Effects 0.000 abstract 2
- 238000001647 drug administration Methods 0.000 abstract 1
- 230000009881 electrostatic interaction Effects 0.000 abstract 1
- 239000003978 infusion fluid Substances 0.000 abstract 1
- 210000004877 mucosa Anatomy 0.000 abstract 1
- 229940098458 powder spray Drugs 0.000 abstract 1
- 239000000243 solution Substances 0.000 description 15
- 239000002245 particle Substances 0.000 description 11
- 201000011510 cancer Diseases 0.000 description 10
- 210000004027 cell Anatomy 0.000 description 10
- 102100032912 CD44 antigen Human genes 0.000 description 9
- 101000868273 Homo sapiens CD44 antigen Proteins 0.000 description 9
- KWYUFKZDYYNOTN-UHFFFAOYSA-M Potassium hydroxide Chemical compound [OH-].[K+] KWYUFKZDYYNOTN-UHFFFAOYSA-M 0.000 description 8
- WYURNTSHIVDZCO-UHFFFAOYSA-N Tetrahydrofuran Chemical compound C1CCOC1 WYURNTSHIVDZCO-UHFFFAOYSA-N 0.000 description 6
- 239000006185 dispersion Substances 0.000 description 6
- 230000000857 drug effect Effects 0.000 description 6
- 230000004048 modification Effects 0.000 description 6
- 238000012986 modification Methods 0.000 description 6
- 229920001223 polyethylene glycol Polymers 0.000 description 6
- 210000004881 tumor cell Anatomy 0.000 description 6
- 230000000259 anti-tumor effect Effects 0.000 description 5
- 231100000135 cytotoxicity Toxicity 0.000 description 5
- 230000003013 cytotoxicity Effects 0.000 description 5
- 235000019441 ethanol Nutrition 0.000 description 5
- 238000011275 oncology therapy Methods 0.000 description 5
- 238000002626 targeted therapy Methods 0.000 description 5
- 238000010521 absorption reaction Methods 0.000 description 4
- 239000002246 antineoplastic agent Substances 0.000 description 4
- 229940041181 antineoplastic drug Drugs 0.000 description 4
- 238000012377 drug delivery Methods 0.000 description 4
- 238000011156 evaluation Methods 0.000 description 4
- 108010003425 hyaluronan-mediated motility receptor Proteins 0.000 description 4
- 229940108949 paclitaxel injection Drugs 0.000 description 4
- 210000002381 plasma Anatomy 0.000 description 4
- 238000005160 1H NMR spectroscopy Methods 0.000 description 3
- 102100027735 Hyaluronan mediated motility receptor Human genes 0.000 description 3
- 102100026849 Lymphatic vessel endothelial hyaluronic acid receptor 1 Human genes 0.000 description 3
- 241000699670 Mus sp. Species 0.000 description 3
- 239000003005 anticarcinogenic agent Substances 0.000 description 3
- 238000012512 characterization method Methods 0.000 description 3
- 201000010099 disease Diseases 0.000 description 3
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 3
- 229960004679 doxorubicin Drugs 0.000 description 3
- 230000014509 gene expression Effects 0.000 description 3
- 238000004128 high performance liquid chromatography Methods 0.000 description 3
- 238000000338 in vitro Methods 0.000 description 3
- 230000002401 inhibitory effect Effects 0.000 description 3
- 229940090044 injection Drugs 0.000 description 3
- 229920001606 poly(lactic acid-co-glycolic acid) Polymers 0.000 description 3
- 230000008569 process Effects 0.000 description 3
- 238000011160 research Methods 0.000 description 3
- 238000007151 ring opening polymerisation reaction Methods 0.000 description 3
- 238000002390 rotary evaporation Methods 0.000 description 3
- YLQBMQCUIZJEEH-UHFFFAOYSA-N tetrahydrofuran Natural products C=1C=COC=1 YLQBMQCUIZJEEH-UHFFFAOYSA-N 0.000 description 3
- IAZDPXIOMUYVGZ-WFGJKAKNSA-N Dimethyl sulfoxide Chemical compound [2H]C([2H])([2H])S(=O)C([2H])([2H])[2H] IAZDPXIOMUYVGZ-WFGJKAKNSA-N 0.000 description 2
- 238000005033 Fourier transform infrared spectroscopy Methods 0.000 description 2
- 101000832213 Homo sapiens Stabilin-2 Proteins 0.000 description 2
- 101710178181 Lymphatic vessel endothelial hyaluronic acid receptor 1 Proteins 0.000 description 2
- 239000002202 Polyethylene glycol Substances 0.000 description 2
- 102100024470 Stabilin-2 Human genes 0.000 description 2
- 238000009825 accumulation Methods 0.000 description 2
- 230000000711 cancerogenic effect Effects 0.000 description 2
- 231100000315 carcinogenic Toxicity 0.000 description 2
- 239000003054 catalyst Substances 0.000 description 2
- 150000001875 compounds Chemical class 0.000 description 2
- 238000013461 design Methods 0.000 description 2
- 239000012738 dissolution medium Substances 0.000 description 2
- 239000000839 emulsion Substances 0.000 description 2
- 239000003292 glue Substances 0.000 description 2
- 230000036541 health Effects 0.000 description 2
- 230000036571 hydration Effects 0.000 description 2
- 238000006703 hydration reaction Methods 0.000 description 2
- 230000001404 mediated effect Effects 0.000 description 2
- 125000002496 methyl group Chemical group [H]C([H])([H])* 0.000 description 2
- KJFMBFZCATUALV-UHFFFAOYSA-N phenolphthalein Chemical compound C1=CC(O)=CC=C1C1(C=2C=CC(O)=CC=2)C2=CC=CC=C2C(=O)O1 KJFMBFZCATUALV-UHFFFAOYSA-N 0.000 description 2
- 229920001553 poly(ethylene glycol)-block-polylactide methyl ether Polymers 0.000 description 2
- 239000000047 product Substances 0.000 description 2
- 230000001681 protective effect Effects 0.000 description 2
- BBEAQIROQSPTKN-UHFFFAOYSA-N pyrene Chemical compound C1=CC=C2C=CC3=CC=CC4=CC=C1C2=C43 BBEAQIROQSPTKN-UHFFFAOYSA-N 0.000 description 2
- 108020003175 receptors Proteins 0.000 description 2
- 102000005962 receptors Human genes 0.000 description 2
- 210000000664 rectum Anatomy 0.000 description 2
- 238000006268 reductive amination reaction Methods 0.000 description 2
- 239000007962 solid dispersion Substances 0.000 description 2
- 241000894007 species Species 0.000 description 2
- 210000002784 stomach Anatomy 0.000 description 2
- 239000006228 supernatant Substances 0.000 description 2
- 210000001519 tissue Anatomy 0.000 description 2
- 238000004448 titration Methods 0.000 description 2
- 231100000331 toxic Toxicity 0.000 description 2
- 230000002588 toxic effect Effects 0.000 description 2
- 238000011282 treatment Methods 0.000 description 2
- DWNBOPVKNPVNQG-LURJTMIESA-N (2s)-4-hydroxy-2-(propylamino)butanoic acid Chemical compound CCCN[C@H](C(O)=O)CCO DWNBOPVKNPVNQG-LURJTMIESA-N 0.000 description 1
- 102000009027 Albumins Human genes 0.000 description 1
- 108010088751 Albumins Proteins 0.000 description 1
- 241000894006 Bacteria Species 0.000 description 1
- DBJPGQWEKGRALK-UHFFFAOYSA-N C(=O)O.CCCCCCCCCCCCCCCCCC Chemical compound C(=O)O.CCCCCCCCCCCCCCCCCC DBJPGQWEKGRALK-UHFFFAOYSA-N 0.000 description 1
- 201000009030 Carcinoma Diseases 0.000 description 1
- 208000017897 Carcinoma of esophagus Diseases 0.000 description 1
- 108010001857 Cell Surface Receptors Proteins 0.000 description 1
- 206010009944 Colon cancer Diseases 0.000 description 1
- 208000001333 Colorectal Neoplasms Diseases 0.000 description 1
- 244000163122 Curcuma domestica Species 0.000 description 1
- 235000003392 Curcuma domestica Nutrition 0.000 description 1
- MWWSFMDVAYGXBV-RUELKSSGSA-N Doxorubicin hydrochloride Chemical compound Cl.O([C@H]1C[C@@](O)(CC=2C(O)=C3C(=O)C=4C=CC=C(C=4C(=O)C3=C(O)C=21)OC)C(=O)CO)[C@H]1C[C@H](N)[C@H](O)[C@H](C)O1 MWWSFMDVAYGXBV-RUELKSSGSA-N 0.000 description 1
- 208000000461 Esophageal Neoplasms Diseases 0.000 description 1
- VGGSQFUCUMXWEO-UHFFFAOYSA-N Ethene Chemical group C=C VGGSQFUCUMXWEO-UHFFFAOYSA-N 0.000 description 1
- 102000002812 Heat-Shock Proteins Human genes 0.000 description 1
- 108010004889 Heat-Shock Proteins Proteins 0.000 description 1
- 101001054921 Homo sapiens Lymphatic vessel endothelial hyaluronic acid receptor 1 Proteins 0.000 description 1
- 206010058467 Lung neoplasm malignant Diseases 0.000 description 1
- 241000699666 Mus <mouse, genus> Species 0.000 description 1
- 238000005481 NMR spectroscopy Methods 0.000 description 1
- 208000002454 Nasopharyngeal Carcinoma Diseases 0.000 description 1
- 206010061306 Nasopharyngeal cancer Diseases 0.000 description 1
- 240000002853 Nelumbo nucifera Species 0.000 description 1
- 235000006508 Nelumbo nucifera Nutrition 0.000 description 1
- 235000006510 Nelumbo pentapetala Nutrition 0.000 description 1
- 208000034530 PLAA-associated neurodevelopmental disease Diseases 0.000 description 1
- 244000131316 Panax pseudoginseng Species 0.000 description 1
- 239000004698 Polyethylene Substances 0.000 description 1
- 108010039918 Polylysine Proteins 0.000 description 1
- 229920002385 Sodium hyaluronate Polymers 0.000 description 1
- HPZOOQSXPMEJBV-ODCFVKFUSA-N Tirilazad mesylate Chemical compound CS(O)(=O)=O.O=C([C@@H]1[C@@]2(C)CC=C3[C@@]4(C)C=CC(=O)C=C4CC[C@H]3[C@@H]2C[C@H]1C)CN(CC1)CCN1C(N=1)=CC(N2CCCC2)=NC=1N1CCCC1 HPZOOQSXPMEJBV-ODCFVKFUSA-N 0.000 description 1
- ZMANZCXQSJIPKH-UHFFFAOYSA-N Triethylamine Chemical class CCN(CC)CC ZMANZCXQSJIPKH-UHFFFAOYSA-N 0.000 description 1
- 239000000853 adhesive Substances 0.000 description 1
- 230000001070 adhesive effect Effects 0.000 description 1
- 239000003513 alkali Substances 0.000 description 1
- 229940037003 alum Drugs 0.000 description 1
- 230000009435 amidation Effects 0.000 description 1
- 238000011938 amidation process Methods 0.000 description 1
- 238000007112 amidation reaction Methods 0.000 description 1
- 238000004458 analytical method Methods 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 238000006065 biodegradation reaction Methods 0.000 description 1
- 230000005540 biological transmission Effects 0.000 description 1
- 238000012661 block copolymerization Methods 0.000 description 1
- 230000017531 blood circulation Effects 0.000 description 1
- 230000037396 body weight Effects 0.000 description 1
- 210000000481 breast Anatomy 0.000 description 1
- 238000004364 calculation method Methods 0.000 description 1
- 208000035269 cancer or benign tumor Diseases 0.000 description 1
- 239000000969 carrier Substances 0.000 description 1
- 230000001413 cellular effect Effects 0.000 description 1
- 230000004700 cellular uptake Effects 0.000 description 1
- 238000002512 chemotherapy Methods 0.000 description 1
- 230000004087 circulation Effects 0.000 description 1
- 230000009514 concussion Effects 0.000 description 1
- 238000013270 controlled release Methods 0.000 description 1
- 238000007334 copolymerization reaction Methods 0.000 description 1
- 239000006184 cosolvent Substances 0.000 description 1
- 239000013078 crystal Substances 0.000 description 1
- 230000001186 cumulative effect Effects 0.000 description 1
- 235000003373 curcuma longa Nutrition 0.000 description 1
- 238000005202 decontamination Methods 0.000 description 1
- 230000003588 decontaminative effect Effects 0.000 description 1
- 230000003111 delayed effect Effects 0.000 description 1
- 238000001514 detection method Methods 0.000 description 1
- 238000011161 development Methods 0.000 description 1
- 102000038379 digestive enzymes Human genes 0.000 description 1
- 108091007734 digestive enzymes Proteins 0.000 description 1
- 238000010790 dilution Methods 0.000 description 1
- 239000012895 dilution Substances 0.000 description 1
- 125000000118 dimethyl group Chemical group [H]C([H])([H])* 0.000 description 1
- 125000002185 docetaxel anhydrous group Chemical group 0.000 description 1
- 229960002918 doxorubicin hydrochloride Drugs 0.000 description 1
- 229940035754 doxorubicin hydrochloride 50 mg Drugs 0.000 description 1
- 239000000890 drug combination Substances 0.000 description 1
- 239000003596 drug target Substances 0.000 description 1
- 230000012202 endocytosis Effects 0.000 description 1
- 210000000981 epithelium Anatomy 0.000 description 1
- LYCAIKOWRPUZTN-UHFFFAOYSA-N ethylene glycol Substances OCCO LYCAIKOWRPUZTN-UHFFFAOYSA-N 0.000 description 1
- 238000001704 evaporation Methods 0.000 description 1
- 230000008020 evaporation Effects 0.000 description 1
- 230000005284 excitation Effects 0.000 description 1
- 210000002950 fibroblast Anatomy 0.000 description 1
- GVEPBJHOBDJJJI-UHFFFAOYSA-N fluoranthrene Natural products C1=CC(C2=CC=CC=C22)=C3C2=CC=CC3=C1 GVEPBJHOBDJJJI-UHFFFAOYSA-N 0.000 description 1
- 210000004907 gland Anatomy 0.000 description 1
- 239000011521 glass Substances 0.000 description 1
- 229930182470 glycoside Natural products 0.000 description 1
- 150000002338 glycosides Chemical class 0.000 description 1
- 238000000703 high-speed centrifugation Methods 0.000 description 1
- 230000005847 immunogenicity Effects 0.000 description 1
- 238000002329 infrared spectrum Methods 0.000 description 1
- 238000011835 investigation Methods 0.000 description 1
- 150000002500 ions Chemical class 0.000 description 1
- 239000002502 liposome Substances 0.000 description 1
- 201000005202 lung cancer Diseases 0.000 description 1
- 208000020816 lung neoplasm Diseases 0.000 description 1
- 210000002540 macrophage Anatomy 0.000 description 1
- 239000000463 material Substances 0.000 description 1
- 238000005259 measurement Methods 0.000 description 1
- 238000005374 membrane filtration Methods 0.000 description 1
- 102000006240 membrane receptors Human genes 0.000 description 1
- 230000036457 multidrug resistance Effects 0.000 description 1
- 230000035772 mutation Effects 0.000 description 1
- 239000002539 nanocarrier Substances 0.000 description 1
- 201000011216 nasopharynx carcinoma Diseases 0.000 description 1
- 239000013642 negative control Substances 0.000 description 1
- 238000000655 nuclear magnetic resonance spectrum Methods 0.000 description 1
- 230000010355 oscillation Effects 0.000 description 1
- 210000001672 ovary Anatomy 0.000 description 1
- 230000002018 overexpression Effects 0.000 description 1
- 239000005022 packaging material Substances 0.000 description 1
- 239000003182 parenteral nutrition solution Substances 0.000 description 1
- 235000010603 pastilles Nutrition 0.000 description 1
- 230000035479 physiological effects, processes and functions Effects 0.000 description 1
- 239000002504 physiological saline solution Substances 0.000 description 1
- 239000002574 poison Substances 0.000 description 1
- 231100000614 poison Toxicity 0.000 description 1
- 239000004632 polycaprolactone Substances 0.000 description 1
- 229920000573 polyethylene Polymers 0.000 description 1
- 229920000656 polylysine Polymers 0.000 description 1
- 238000006116 polymerization reaction Methods 0.000 description 1
- 239000013641 positive control Substances 0.000 description 1
- IQVVSTXAINGURX-UHFFFAOYSA-N propan-2-one;pyrene Chemical compound CC(C)=O.C1=CC=C2C=CC3=CC=CC4=CC=C1C2=C43 IQVVSTXAINGURX-UHFFFAOYSA-N 0.000 description 1
- ZAHRKKWIAAJSAO-UHFFFAOYSA-N rapamycin Natural products COCC(O)C(=C/C(C)C(=O)CC(OC(=O)C1CCCCN1C(=O)C(=O)C2(O)OC(CC(OC)C(=CC=CC=CC(C)CC(C)C(=O)C)C)CCC2C)C(C)CC3CCC(O)C(C3)OC)C ZAHRKKWIAAJSAO-UHFFFAOYSA-N 0.000 description 1
- 230000009467 reduction Effects 0.000 description 1
- 238000006722 reduction reaction Methods 0.000 description 1
- 230000002829 reductive effect Effects 0.000 description 1
- 238000012827 research and development Methods 0.000 description 1
- 239000012313 reversal agent Substances 0.000 description 1
- 239000012266 salt solution Substances 0.000 description 1
- 238000007789 sealing Methods 0.000 description 1
- 239000013049 sediment Substances 0.000 description 1
- 238000000926 separation method Methods 0.000 description 1
- 239000012679 serum free medium Substances 0.000 description 1
- 229960002930 sirolimus Drugs 0.000 description 1
- QFJCIRLUMZQUOT-HPLJOQBZSA-N sirolimus Chemical compound C1C[C@@H](O)[C@H](OC)C[C@@H]1C[C@@H](C)[C@H]1OC(=O)[C@@H]2CCCCN2C(=O)C(=O)[C@](O)(O2)[C@H](C)CC[C@H]2C[C@H](OC)/C(C)=C/C=C/C=C/[C@@H](C)C[C@@H](C)C(=O)[C@H](OC)[C@H](O)/C(C)=C/[C@@H](C)C(=O)C1 QFJCIRLUMZQUOT-HPLJOQBZSA-N 0.000 description 1
- 229940010747 sodium hyaluronate Drugs 0.000 description 1
- YWIVKILSMZOHHF-QJZPQSOGSA-N sodium;(2s,3s,4s,5r,6r)-6-[(2s,3r,4r,5s,6r)-3-acetamido-2-[(2s,3s,4r,5r,6r)-6-[(2r,3r,4r,5s,6r)-3-acetamido-2,5-dihydroxy-6-(hydroxymethyl)oxan-4-yl]oxy-2-carboxy-4,5-dihydroxyoxan-3-yl]oxy-5-hydroxy-6-(hydroxymethyl)oxan-4-yl]oxy-3,4,5-trihydroxyoxane-2- Chemical compound [Na+].CC(=O)N[C@H]1[C@H](O)O[C@H](CO)[C@@H](O)[C@@H]1O[C@H]1[C@H](O)[C@@H](O)[C@H](O[C@H]2[C@@H]([C@@H](O[C@H]3[C@@H]([C@@H](O)[C@H](O)[C@H](O3)C(O)=O)O)[C@H](O)[C@@H](CO)O2)NC(C)=O)[C@@H](C(O)=O)O1 YWIVKILSMZOHHF-QJZPQSOGSA-N 0.000 description 1
- 238000000527 sonication Methods 0.000 description 1
- 230000006641 stabilisation Effects 0.000 description 1
- 238000011105 stabilization Methods 0.000 description 1
- 238000003756 stirring Methods 0.000 description 1
- 230000004083 survival effect Effects 0.000 description 1
- 238000010189 synthetic method Methods 0.000 description 1
- 230000009885 systemic effect Effects 0.000 description 1
- 125000002456 taxol group Chemical group 0.000 description 1
- 231100000419 toxicity Toxicity 0.000 description 1
- 230000001988 toxicity Effects 0.000 description 1
- 230000005909 tumor killing Effects 0.000 description 1
- 230000005760 tumorsuppression Effects 0.000 description 1
- 235000013976 turmeric Nutrition 0.000 description 1
- 238000002604 ultrasonography Methods 0.000 description 1
- 210000003462 vein Anatomy 0.000 description 1
- 238000005406 washing Methods 0.000 description 1
Landscapes
- Medicinal Preparation (AREA)
- Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
Abstract
本发明属于高分子化学和药物制剂领域。具体涉及主动靶向透明质酸‑聚乳酸载体的合成及其抗肿瘤药物胶束的制备方法和应用;采用新型自组装技术通过静电作用,将两亲性的PEG嵌段聚酯共聚物与肿瘤靶向性配体透明质酸‑聚乳酸共聚物自组装形成多功能复合胶束,该载抗癌药胶束与复合胶束组合物,能显著提高难溶性肿瘤药溶解性或水溶性抗癌药的载药量与包封率,体内能生物降解,能够避免网状内皮系统(RES)的吞噬和肾脏的排泄,具有长循环作用,多功能组合物突出优势有肿瘤主动靶向作用,体内外药效学参数明显优于抗肿瘤药普通注射剂。临床上可接受的给药途径有注射给药或粘膜给药;剂型为注射剂、输液剂、注射冻干粉针剂或粉雾剂。
Description
技术领域
本发明属于高分子化学和药物制剂领域。具体涉及主动靶向透明质酸-聚乳酸载体的合成及其抗肿瘤药物胶束的制备方法和应用;本发明还涉及具有主动靶向生物降解载体与两亲性嵌段共聚物载体自组装新技术。
背景技术
癌症严重是危害人们健康和生命的常见病,全球每年新发病1200万,死亡率高,我国每年发病人数超过220万,恶性肿瘤的发病死亡率在我国逐年上升已是不争的事实,并已成为造成我国居民疾病死亡的首要病因。据国家卫生部信息中心的报道,近年来危害我国民众的主要癌症类型依次为胃癌(21.76%)、肝癌(17.83%)、肺癌(15.19%)、食管癌(15.02%)、结直肠癌(4.54%)、白血病(3.53%)、子宫颈癌(1.64%)、鼻咽癌(1.53%)和乳腺癌(1.49%)。
目前癌症的临床治疗仍以化疗为主,上市的紫杉醇和多西紫杉醇普通注射剂的毒副作用大,缺点是化疗药物对癌细胞没有选择性,会导致相当数量正常细胞的死亡,热点是针对特定的癌症类型、特定病人研究开发更有针对性、副作用更少“靶向治疗”。抗肿瘤药物新型制剂得以研究和开发。其中紫杉醇脂质体(力朴素)、白蛋白纳米粒已上市并应用与临床。
EP0583955B1专利公开一种单甲基聚乙二醇-聚乳酸载多西紫杉醇胶束的制备方法,将单甲基聚乙二醇聚乳酸和多西紫杉醇,在60℃水浴溶解于乙腈,将乙腈液缓慢滴加到纯化水中,搅拌1h,60℃常压蒸发5h,胶束粒径为50nm,载药量为6%。Kim等采用PEO-b-PDLLA制备-PM胶束,已进入临床研究阶段,然而该胶束存在药物消除速率过快的问题,剂量提高2.5倍,该胶束血浆的AUC为Taxol的74.1%,血浆清除速率CL为Taxol的3倍[Kim S C,Kim D W,Shim Y H,et al.In vivo evaluation of polymeric micellarpaclitaxel formulation:toxicity and efficacy.Journal of Controlled Release,2007,72(1):191-202]。Glen S.Kwon等授权一种肿瘤多药耐药逆转剂热休克蛋白(Hsp-90),采用Hsp-90修饰PEG-b-PLA包载紫杉醇或雷帕霉素,薄膜旋转蒸发法制备胶束,过滤除菌冷冻干燥即得载药胶束冻干粉,该胶束存在主要问题,A549荷瘤小鼠药效学评价中,胶束组de1药效远低于市售制剂组[US8529917]。
针对被动靶向载体嵌段共聚物胶束存在体内靶向性低问题,提出了靶向治疗,是指在细胞分子水平上,针对已经明确的致癌位点,来设计相应的治疗药物,药物进入体内会特异地选择致癌位点来相结合发生作用,使肿瘤细胞特异性死亡,而不会波及肿瘤周围的正常组织细胞。通过靶向治疗可以改善药物在肿瘤部位的分布,上皮细胞癌、卵巢癌、直肠癌、胃癌等多种肿瘤细胞表面阳性表达透明质酸受体CD44,RHAMM、HARE和LYVE-1等,表达量高,而正常细胞CD44、RHAMM受体阴性。
透明质酸(HA)在人体内存在多种特异性受体,如CD44,RHAMM、HARE和LYVE-1等,而CD44、RHAMM在多种肿瘤(如上皮、卵巢、直肠、胃部肿瘤)均有过度表达。HA具有高度特异性和生物相容性,HA修饰载体是设计和制备聚合物胶束亲水链段介导肿瘤主动靶向成的关键。将药物主动靶向肿瘤细胞,从而提高药物在肿瘤组织分布,达到肿瘤靶向治疗目的。
Choi K Y等合成PEG修饰的透明质酸-5β-胆烷酸聚合物,将聚合物溶于水相,喜树碱DMSO溶液缓慢滴入聚合物水相,用透析法制备胶束。与成纤维细胞NIH-3Y3相比,在CD44过表达SCC7癌细胞,细胞摄取量显著增加。但该方法缺点是胶束的平均粒径较大为319.6nm,另透明质酸包覆于PEG内部无法暴露,影响与靶细胞表面受体的作用,使药物难以从网状内皮系统(RES)逃逸,被单核巨噬细胞系统(MPS)清除,降低了胶束中药物在肿瘤部位蓄积[Choi K Y,Yoon H Y, Kim J-H,et al.,Smart nanocarrier based on PEGylatedhyaluronic acid for cancer therapy.ACS nano,2011,5(11):8591-9]。Pitarresi G合成透明质酸接枝聚乳酸与聚乙二醇(HA-g-PLA-g-PEG)共聚,制备阿霉素胶束,该胶束通过CD44受体介导内吞作用增加HCT116细胞的摄取,但PLA接枝率较低为5.0%,导致该胶束载药量低为4.8%,包封率为9.6%,抗肿瘤药效降低[Pitarresi G, Palumbo F S,AlbaneseA,et al.,Self-assembled amphiphilic hyaluronic acid graft copolymers fortargeted release of antitumoral drug.J Drug Target,2010,18(4):264-76]。
Yadav A K等制备阿霉素(DOX)HA-PEG-PLGA胶束,将PLGA-PEG-NH2溶于纯化水中加入HA和EDCA,磁力搅拌7h,冷冻干燥得HA-PEG-PLGA。用沉淀分散法水相方法制备聚合物胶束,包封率为87.66%。药动学结果表明,DOX胶束血浆药物浓度远低于游离DOX,可能由于血浆中抗透明质酸酶和玻尿酸降解酶的作用分解胶束,降低药物在血液循环滞留时间[Yadav A K,Mishra P,Mishra A K,et al.,Development and characterization ofhyaluronic acid-anchored PLGA nanoparticulate carriers ofdoxorubicin.Nanomedicine:nanotechnology,biology,and medicine,2007,3(4):246-57]。中国专利201010228961授权一种透明质酸-C18胶束的制备方法,将HA-C18和紫杉醇溶于DMSO中,磁力搅拌混合均匀,用透析法透析除有机溶剂和游离紫杉醇,粒径为236.6nm,载药量为9.57%,与聚酯相比十八烷甲酸的生物相容性不好,今后临床应用有困难。
基于HA与聚酯两亲性生物降解载体修饰药物传递系统,对于一些肿瘤治疗具有重大潜在应用价值,新型给药系统对抗癌药临床应用提供理论依据。本发明以透明质酸为主动靶头通过与聚酯合成系列两嵌段共聚物,以透明质酸为基础构建的主动靶向药物传递系统,具有免疫原性低、生物相容性良好和可降解等优点,还具有低临界胶束浓度和药物缓释等特性;HA与CD44高表达的肿瘤细胞特异性结合,引导纳米胶束在肿瘤细胞中主动蓄集,达到主动靶向肿瘤细胞目的。
发明内容
本发明涉及主动靶向透明质酸-聚乳酸载体的合成及其抗肿瘤药物胶束的制备方法和应用;本发明还涉及具有主动靶向生物降解载体与两亲性嵌段共聚物载体自组装新技术。本发明的目的是将透明质酸-聚酯(聚乳酸、聚己内酯等)通过还原胺化反应合成,采用该两亲性载体包覆抗癌药物制备聚合物胶束;更重要的目的是采用自组装技术通过静电作用,将两亲性的PEG嵌段聚酯共聚物与肿瘤靶向性配体透明质酸-聚乳酸共聚物自组装形成多功能复合胶束。
本发明利用透明质酸分子中含有多种基团通过共价键与聚酯中活性基团结合,通过酰氨化反应合成HA-聚酯,通过与PEG化-聚酯载体自组装新技术新策略,使PEG在循环过程中屏蔽HA表面,通过摄取前暴露HA靶点,更有利于延长药物在循环系统中的循环时间,其双亲水外壳和聚酯环绕的疏水内核,可逃避单核巨噬细胞的吞噬,减少药物全身性扩散的毒副作用提高抗癌药物靶向治疗作用。
本发明提供一种或系列具有肿瘤靶向性载体和多功能作用的自组装复合胶束组合物,其特征在于:透明质酸末端的羟基与聚乳酸中羧基通过共价键结合,为具有两亲性的嵌段共聚物,其结构通式(I):
其中透明质酸分子量为4500~45000Da,聚乳酸分子量为1200~13000Da,m范围为20~92,n范围为30~103;
所述(I)的透明质酸-聚乳酸抗癌药物自发形成胶束,通过自组装新技术与AB、ABA、ABC两亲性-聚酯嵌段共聚物包覆抗癌药物成多功能自组装胶束,其自组装胶束由双亲水性外壳和聚酯环绕的疏水内核组成;
所述透明质酸与聚乳酸的重量比为1∶0.2~0.8;透明质酸-聚乳酸与两亲性聚酯共聚物的自组装胶束组合物的重量比为:1∶0.2~4.0;
所述的透明质酸-聚乳酸,可以包载疏水性或亲水性抗肿瘤药物,其中药物与(I)的重量比为1∶1.0~200。
本发明提供一种主动靶向透明质酸-聚乳酸载体及其抗肿瘤药物胶束组合物,其特征在于所述难溶性抗肿瘤药物选自紫杉醇、多烯紫杉醇、阿霉素、冬凌草素、姜黄素、姜黄素3、人参皂苷Rh2、柔红霉素、表柔比星、10羟基喜树碱、9-硝基喜树碱、S-38、甲氨蝶呤、依托泊苷、米托蒽醌或其衍生物;水溶性抗肿瘤药物选自有盐酸阿霉素、盐酸拓扑霉素、盐酸柔红霉素等。
本发明的透明质酸-聚乳酸共聚物载体的合成方法包括如下步骤:
(1)聚乳酸的合成
将D,L-丙交酯适量加入圆底烧瓶中,油浴温度为100-135℃,待丙交酯熔融,加入催化剂Sn(Oct)2,通氮气排净容器中空气,逐步升温至140-175℃温度,聚合反应6-8h,加有机溶剂a纯化,真空干燥即得PLA固体;
聚乳酸的活化:将聚乳酸溶于有机溶剂a,加N,N-二环己基碳二亚胺和N-羟基-琥珀酰亚胺,氮气保护,室温反应24h,用有机溶剂b沉淀,抽滤,真空干燥,即得PLA-NHS;
(2)透明质酸-聚乳酸的合成
透明质酸的末端胺化合成:将N,N-二乙基乙二胺用pH7-9.5磷酸盐缓冲溶解,称取透明质酸加纯化水溶解滴入上述溶液中,磁力搅拌2-6h。加还原试剂,控制温度8~20℃反应1h,室温反应12h;将反应物放入透析袋中,用PBS缓冲溶液透析两天,冷冻干燥冻干,即得末端胺化-透明质酸HA-NH2;
称取HA-NH2适量,加pH7-9.5PBS缓冲溶液,超声溶解,加PLA-NHS有机溶剂,室温磁力搅拌反应15-30h,采用有机溶剂a,除去杂质沉淀;纯化水透析袋法透析24-72h,除去未反应的HA-NH2,冷冻干燥,即得HA-PLA固体;
其中所述有机溶剂a为甲醇、乙醇、乙醚、乙腈、氯仿、二氯甲烷、二甲基亚砜一种或混合物;所述溶剂b为石油醚、冰乙醚、乙酸乙酯的一种或混合物;
所述还原试剂选自氰基硼氢化钠、硼氢化钠等。
本发明的具有主动靶向透明质酸-聚乳酸载体及自组装多功能胶束组合物,其特征在于,所述两亲性-聚酯为AB、ABA或ABC型两嵌段共聚物、三嵌段共聚物或接枝共聚物,其中两嵌段共聚物选自mPEG-PLA、mPEG-PCL、mPEG-PLGA,三嵌段共聚物选自mPEG-PLLA-PMMD、PLA-PEG-PLA、PCL-PEG-PCL或PLGA-PEG-PLGA;PMMD为聚丙氨酸、聚赖氨酸、聚甘氨酸、聚谷氨酸,m为800~20000。
本发明的具有主动靶向透明质酸-聚乳酸载体及其抗肿瘤药物胶束组合物的制备方法,其特征在于,所述聚合物胶束和复合胶束组合物的制备方法如下:
(1)聚合物胶束的制备:称取透明质酸-聚乳酸溶于纯化水,抗肿瘤药物加有机溶剂a超声溶解,将含药相滴入载体水相,置恒温磁力搅拌仪,扩散2-6h得胶束初混悬液;再将上述胶束初混悬液移至透析袋中,在纯化水或磷酸盐缓冲液中透析12-48h,探头超声5-40min,0.22μm微孔滤膜过滤,即得胶束混悬液;另取胶束混悬液,加冻干保护剂,分装西林瓶冷冻干燥,即得胶束固体粉;或
(2)复合胶束组合物的制备:两亲性聚酯溶解于有机溶剂,抗肿瘤药物溶于有机溶剂;将含药相缓慢滴入载体有机相,置恒温磁力搅拌仪,扩散2-6h即得胶束初混悬液;再将上述胶束初混悬液置入旋转蒸发仪发挥除去有机溶剂,加纯化水分散,即得胶束混悬液1;另取两亲性聚酯载体加有机溶剂超声溶解,滴入磁力搅拌的胶束混悬液1中,固体分散组装2~3h,探头超声5-30min,0.22μm微孔滤膜过滤,即得自组装复合胶束混悬液;另取自组装复合胶束混悬液,加冻干保护剂,分装西林瓶冷冻干燥,即得自组装复合胶束固体粉。
本发明所提供的主动靶向透明质酸-聚乳酸载体及其抗肿瘤药物胶束组合物,其特征是,其中所述有机溶剂为甲醇、乙醇、乙腈、丙酮、乙醚、二氯甲烷、三氯甲烷等的一种或两种混合溶剂;
所述聚合物胶束或复合胶束冻干保护剂选自甘露醇、葡萄糖、乳糖、蔗糖或海藻糖中的一种或几种,冻干保护剂用量按载药聚合物胶束混悬液重量计,1份载药聚合物胶束混悬液加入冻干保护剂0.01~0.2份。
本发明所提供的主动靶向透明质酸-聚乳酸载体及其抗肿瘤药物胶束组合物,其特征是,该组合物具有改善抗癌药溶解性,载药量大于5%,平均粒径小于250nm,制剂稳定性好。体内能生物降解,该载药胶束与复合胶束,能够避免网状内皮系统(RES)的吞噬和肾脏的排泄,具有长循环作用和肿瘤主动靶向作用,体内外药效学明显高于抗肿瘤药普通注射剂。
本发明所提供的主动靶向透明质酸-聚乳酸载体及其抗肿瘤药物胶束组合物,其特征在于该载药胶束和复合胶束可用于治疗非小细胞肺癌、乳腺癌、卵巢癌、子宫颈癌、胃癌、肝癌、白血病、肾癌等恶性肿瘤的单一或联合用药。
附图说明
图1是透明质酸-聚乳酸-紫杉醇聚合物胶束与自组装胶束的释放曲线(n=6)
图2是紫杉醇注射剂、自组装胶束和透明质酸-聚乳酸紫杉醇胶束HepG-2细胞毒性图(n=3)
具体实施方式
实施例1 聚乳酸(PLA)的合成及其活化
聚乳酸(PLA)的合成:采用开环聚合法,将10g D,L-丙交酯(69.4mmol)加入单口圆底烧瓶中,油浴温度为120℃,待丙交酯熔融,加入催化剂1.8mL Sn(Oct)2(5mmol)。通氮气排净容器中空气,逐步升温至160℃温度,聚合反应6h。加入乙酸乙酯适量溶解,再加乙醇沉淀,抽滤,离子水洗涤,真空干燥即得淡黄色PLA固体,收率为80.2%。开环聚合反应温度与时间对聚乳酸分子量的影响结果见表1。
聚乳酸反应式如下:
采用端基滴定法测定PLA分子量。配制二氯甲烷-无水乙醇(v/v,1∶1)聚乳酸溶液(浓度为100mg/mL),用0.02mol/L氢氧化钾乙醇液滴定终点,以酚酞为指示剂。聚乳酸的重均分子量为计算公式:Mn=1000m/℃(V-V0),m为PLA质量(g);C为氢氧化钾的摩尔浓度(mol/L);V为聚合物耗氢氧化钾溶液体积(mL);V0为空白溶剂耗氢氧化钾溶液体积(mL)。
表1 开环聚合反应温度与时间对聚乳酸分子量的影响
聚乳酸(PLA3500)活化:采用酰胺化反应,将5.3g PLA3500(1.5mmol)溶于二氯甲烷40mL,依次加618.9mgN,N-二环己基碳二亚胺(DCC,3.0mmol)和345.3mgN-羟基-琥珀酰亚胺(NHS)(3.0mmol)(PLA/DCC/NHS摩尔比为1∶2∶2),氮气保护,室温反应24h。用冰乙醚沉淀,静置抽滤,真空干燥4h,即得NHS修饰PLA(PLA3500-NHS),收率为86.65%,
PLA--NHS合成反应式如下:
实施例2 透明质酸-聚乳酸(HA-PLA)的合成
(1)透明质酸的末端胺化合成
透明质酸(Mn:7800)胺化:采用还原胺化反应,将150mg N,N-二乙基乙二胺(1.3mmol)用pH9.5磷酸盐缓冲溶解,称取9.5g透明质酸(HA7800)加纯化水100mL溶解(1.2mmol)滴入上述溶液中,磁力搅拌4h。加0.04g氰基硼氢化钠(0.6mmol),控制温度8~20℃反应1h,室温反应12h。将反应物放入透析袋中(截留分子量为8000~10000),再用PBS缓冲溶液透析两天。冷冻干燥冻干,即得末端胺化-透明质酸(HA-NH2),收率85.91%。
(2)HA7800-PLA3500的合成
8.2g称取HA-NH2(1.1mmol),加PBS(pH9.5)缓冲溶液100mL,超声溶解,加入4.33gPLA3500-NHS(1.2mmol)DMSO溶液(HA与PLA质量比为2∶1),室温磁力搅拌反应24h。加甲醇适量,除去未反应PLA-NHS沉淀。透析袋中用纯化水透析48h除去未反应的HA-NH2,冷冻干燥,即得HA7800-PLA3500固体,产率为71.2%。根据具体要求,通过调整合成条件和HA与PLA质量比,合成反应获得以下产物HA35000-PLA3500、HA7800-PLA6000和HA35000-PLA6000,产率分别为59.7%、77.2%。和61.4%。
HA-PLA的合成步骤如下式:
实施例3 透明质酸-聚乳酸的结构表征和性质测定
(1)结构表征具体步骤
红外光谱(FT-IR)表征:取PLA、HA和HA-PLA适量,与KBr混匀研成粉末,压制成薄片,采用NICO LET-5700型傅立叶红外光谱仪测定与分析。从HA-PLA的FT-IR光图结果分析如下:HA特征吸收峰为3424.5cm-1(υasOH和υasNH),2892.8cm-1(υC-H),1628.7cm-1(υasC-O);PLA特征吸收峰为1758.1cm-1(υC=O),2995.8cm-1(υasCH3),1455.6cm-1(δasCH3);2892.8cm-1处峰强度加大,通过以上结果分析,证实PLA以新生成的酰胺键与HA共价连接,确定合成产物为透明质酸-聚乳酸(HA-PLA)嵌段共聚物。
核磁共振光谱(1H NMR)表征:取HA-PLA适量,溶于DMSO-d6,300MHz频率,采用AV-III-500型核磁共振仪进行测定与分析。1H NMR表征HA-PLA结构,HA-PLA的1H-NMR图中观察到,PLA特征吸收峰为δ1.32和δ1.38(d,3H,-O-CO-CH(CH 3)-O-),δ5.18(m,-O-CO-CH(CH3)-O-)和HA的特征吸收峰:δ1.85(s,3H,-NH-CO-CH 3);δ3.32~3.58(m,-OH);δ3.25(t,2H,-N-CH2-CH 2-O-)。上述分析可知,PLA成功共价键连接到透HA上,成功合成两嵌段共聚物HA-PLA。
(2)嵌段共聚物的CMC测定
采用芘荧光探针法测定HA7800-PLA3500和HA7800-PLA6000两种载体CMC。取芘-丙酮溶液(5×10-5mol/L)10μL置离心管,挥干丙酮,分别系列加入浓度为0.05、0.1、0.2、0.4、1、2、4、10、20、40、100、200、400、1000和2000μg/mL HA-PLA液,芘浓度为5×10-7mol/L。激发波长为392nm,发射波长为338nm和333nm,激发缝宽和发射缝宽分别为5nm和3nm。以I338/I333对浓度对数值作图,发生突变处对应的浓度即为临界胶束浓度。结果显示,两种载体CMC分别为1.985×10-2g/L和2.41×10-3g/L。
(3)包封率与载药量测定
采用高速离心法测定空白与载药胶束的包封率与载药量。取紫杉醇或抗癌药胶束/复合胶束适量高速离心,转速为8000rpm,离心10min,取上清液1mL,加甲醇稀释,吸取25μL注入HPLC测定药物浓度,计算包封率和载药量,HPLC色谱条件:流动相:甲醇∶水(70∶30);流速:1.0mL/min;进样量:20μL;检测波长:227nm;柱温:30℃。
实施例4 乳化-溶剂扩散与透析结合法制备紫杉醇胶束
称取200mg HA7800-PLA6000溶于纯化水25mL,紫杉醇加二甲基亚砜(DMSO)适量超声溶解(投药量为5%、10%和15%),将含药相滴入载体水相,置恒温磁力搅拌仪,扩散2h得胶束初混悬液。再将上述胶束初混悬液移至透析袋中(MWCO为12000),在纯化水中透析12h,除去杂质与有机溶剂,探头超声5min,0.22μm微孔滤膜过滤,即得胶束混悬液,不同投药量对胶束粒径和包封率的影响见表2。另用透射电镜检测胶束的形态,胶束为圆整球形,采用Zetasizer3000激光粒度分析仪测定胶束粒径分布、分散系数和Zeta电位,其平均粒径为150nm,Zeta电位为-10.7mv载药量为4~10%;包载紫杉醇胶束嵌段共聚物的种类对其理化性质影响,结果见表3。
表2 紫杉醇胶束投药量与理化性质的影响(n=3)
表3 包载紫杉醇胶束嵌段共聚物种类对理化性质影响(n=3)
实施例5 乳化/溶剂挥发-薄膜分散法-固体分散法自组装紫杉醇复合胶束
称取200mg HA7800-PLA6000溶于氯仿35mL,称取紫杉醇适量溶于乙腈。将含药相缓慢滴入载体有机相,置恒温磁力搅拌仪,扩散3h得胶束初混悬液;再将上述胶束初混悬液置入旋转蒸发仪发挥除去氯仿,加纯化水分散,即得胶束混悬液1;另取两亲性聚酯载体250mgmPEG-PLA-PMMD(2∶2∶3)加丙酮适量超声溶解,滴入磁力搅拌的胶束混悬液1中,固体分散法组装2~3h,探头超声10min,0.22μm微孔滤膜过滤,即得自组装复合胶束混悬液。另取自组装复合胶束混悬液50mL,加冻干保护剂8.0g甘露醇,分装西林瓶冷冻干燥,即得自组装复合胶束固体粉。
采用Zetasizer3000激光粒度分析仪测定自组装复合胶束粒径分布、分散系数和Zeta电位,复溶于水的自组装复合胶束的平均粒径为205.6nm,Zeta电位为2.56mv,载药量为7.23%,包封率为90.5%。
实施例6 微相分离法制备多西紫杉醇胶束
称取200mg HA35000-PLA3500和30mg多西紫杉醇加入混合溶剂(混合溶剂由2g液体聚乙二醇600和1.8g乙醇作为助溶剂)制备一种混合物。缓慢加热该混合物至60℃,磁力搅拌45min,冷却至室温,,即得多西紫杉醇胶束组合物,该组合物用纯化水稀释,置透析袋(MWCO:12000)在纯化水中透析12h,除去聚乙二醇600,用0.45μm微孔滤膜过滤,即得胶束混悬液。测定平均粒径为286.7nm,Zeta电位为-20.4mv,载药量为9.04%,包封率为84.2%。
实施例7 10-羟基喜树碱自组装胶束的制备
称取两嵌段聚合物120mg mPEG2000-PCL(6∶4)置茄形瓶中,加丙酮20mL,超声溶解,40℃水旋转蒸发除去有机溶剂,加纯化水适量水合,即得mPEG-PCL空白胶束溶液;称取100mg HA-PLA和20mg10-羟基喜树碱溶于四氢呋喃(THF)20mL,超声溶解;另取mPEG-PCL空白胶束溶液适量,与上述含药溶液混合置玻璃离心管中,密封剧烈震荡形成乳浊液。室温缓磁力搅拌24h,将该乳浊液置透析袋(MWCO:12000)于纯化水中透析24h,除去THF,经0.45μm微孔滤膜过滤,即得10-羟基喜树碱自组装胶束混悬液。另取该自组装胶束混悬液适量,加冻干保护乳糖10.5g,混合混匀,经冷冻干燥,即得自组装胶束冻干粉固体。
采用Zetasizer3000激光粒度分析仪测定自组装复合胶束粒径分布、分散系数和Zeta电位,冻干粉复溶于水自组装复合胶束,测定平均粒径为210.6nm,Zeta电位为-21.6mv,载药量为5.83%,包封率为79.8%。
实施例8 阿霉素自组装胶束的制备
取两亲性嵌段聚合物100mg mPEG5000-PLA(7∶3)置于茄形瓶中,加入丙酮20mL,超声溶解,40℃旋转蒸发除去有机溶剂,加纯化水适量水合,即得mPEG5000-PLA空白胶束液1,作为水相;取盐酸阿霉素50mg溶于DMSO适量,滴加36.5mg三乙胺,避光下磁力搅拌过夜,即得阿霉素溶液。称取120mg HA7800-PLA6000加DMSO适量溶解,另取上述载体溶液与阿霉素溶液混匀,即得阿霉素胶束液2;另取mPEG5000-PLA空白胶束液1与阿霉素胶束液2混合均匀呢,室温磁力缓慢搅拌24h,进行自组装复合胶束的包覆,将复合胶束移至透析袋(MWCO:12000)于纯化水中透析24h,除去DMSO,探针超声10min,经0.45μm微孔滤膜过滤,即得自组装复合胶束混悬液。取该自组装胶束混悬液适量,加冻干保护海藻糖7.8g,混合混匀,经冷冻干燥,即得自组装胶束冻干粉固体。
采用Zetasizer3000激光粒度分析仪测定自组装复合胶束粒径分布、分散系数和Zeta电位,冻干粉复溶于水自组装复合胶束。测定平均粒径为175.2nm,Zeta电位为-7.5mv,载药量为6.38%,包封率为89.7%。
实施例9 体外释放研究
取HA-PLA/PTX胶束和HA-PLA/mPEG-PLA-PMMD/PTX自组装胶束混悬液0.5mL(浓度为1.0mg/mL)装入透析袋(MWCO=8000Da),释放介质pH7.4PBS缓冲溶液(含Tween800.5%(v/v))100mL,于37℃恒温振荡水浴仪,进行上述两种含药胶束制剂的体外释放研究。分别于15、30min和1、2、4、6、8、12、24、36和48h取样1mL,补充等量释放介质,经0.45μm滤膜过滤,吸取50μL注入HPLC,色谱条件同实施例3,测定紫杉醇浓度,计算累计释放量。结果见图1,HA-PLA/PTX胶束24h、48h累计释放度分别为54.91%和71.85%,而自组装胶束24h、48h累计释放度分别为58.65%和75.26%,结果表明两种紫杉醇胶束具有缓慢释药作用。
实施例10 体外抗肿瘤药效与细胞毒性研究
取对数生长期的HepG-2细胞接种于96孔,培养24h后,每孔分别加入相应浓度药物(PTX溶液剂、HA-PLA/PTX和HA-PLA/mPEG-PLA-PMMD/PTX自组装复合胶束)无血清培养基200μL,各组样品紫杉醇浓度分别为0.01、0.1、1、10、20、50和100μg/mL。每组设定三个复孔,孵育48h,加入浓度为5mg/mLMTT20μL,继续孵育4h,吸除孔内上清液,加二甲基亚矾150μL溶解紫色结晶,于酶标仪490nm处测定样品的吸收度(OD)。计算相对细胞存活率和IC50,评价靶向胶束的细胞毒性。
由图2细胞毒性结果显示,体外浓度从10μg/mL抑瘤效果比PTX溶液组高,HA-PLA/PTX胶束和HA-PLA/mPEG-PLA-PMMD/PTX复合胶束组IC50值分别为28.28μg/mL和10.21μg/mL相比于PTX组(45.29μg/mL)显著降低(p<0.05),表明靶头HA修饰的紫杉醇可使胶束通过CD44受体介导的胞吞作用大量进入细胞;HA-PLA紫杉醇胶束体外抗肿瘤活性增高,HA-PLA/mPEG-PLA-PMMD/PTX复合胶束有较强的细胞毒性,其体外抗肿瘤药效显著增强。
实施例11 体内抗肿瘤药效的研究
建立Heps荷瘤小鼠为模型,随机将Heps-荷瘤小鼠随机分成4组:阴性对照组生理盐水,阳性对照组(市售紫杉醇注射液);HA-PLA/PTX胶束组和HA-PLA/mPEG-PLA-PMMD/PTX自组装胶束组(给药计量均为10mg/kg)。每组5只,分别于0天、2天、4天和6天尾静脉注射给药。每天测量体重,于第7天将小鼠处死,剥取分离肿瘤,称瘤重,测量瘤体积,计算抑瘤率,进行紫杉醇胶束和自组装胶束靶向制剂的药效学评价。结果见表4,由表4显示,HA-PLA/PTX荷瘤肿瘤抑制药效学研究。结果表明,与紫杉醇注射液相比,构建透明质酸-聚乳酸紫杉醇胶束与自组装多功能复合胶束靶向给药系统,以HA为配体,具有CD44介导主动靶向肿瘤作用。体内药效学评价表明,HA-PLA/PTX胶束组体积抑瘤率为53.96%,自组装复合胶束组的体积抑瘤率为69.72%,均高于注射液40.35%。胶束与自组装多功能复合胶束载药胶束,前者可增强肿瘤主动靶向性,后者还具有长循环作用,显著降低毒副作用,均可提高药效。
表4 不同紫杉醇制剂的药效学结果(n=5)
*与生理盐水组相比显著性差异,P<0.05
Δ与紫杉醇注射液组相比有显著性差异,P<0.05。
Claims (4)
1.一种主动靶向透明质酸-聚乳酸载体,其特征在于:透明质酸末端的羟基胺化,与聚乳酸中羧基通过酰胺键结合,为具有两亲性的嵌段共聚物载体,其中透明质酸分子量为4500~45000Da,聚乳酸分子量为1200~13000Da,m范围为20~92,n范围为30~103;所述透明质酸与聚乳酸的重量比为1∶0.2~0.8,其结构通式(I):
所述透明质酸-聚乳酸共聚物(I)载体的制备方法包括如下步骤:
(1)聚乳酸的合成
将D,L-丙交酯适量加入圆底烧瓶中,油浴温度为100-135℃,待丙交酯熔融,加入催化剂Sn(Oct)2,通氮气排净容器中空气,逐步升温至140~175℃温度,聚合反应6-8h,加有机溶剂a纯化,真空干燥,即得聚乳酸固体;
聚乳酸的活化:将聚乳酸溶于有机溶剂a,加N,N-二环己基碳二亚胺和N-羟基-琥珀酰亚胺,氮气保护,室温反应24h,用有机溶剂b沉淀,抽滤,真空干燥,即得聚乳酸-NHS;
(2)透明质酸-聚乳酸的合成
透明质酸的末端胺化合成:将N,N-二乙基乙二胺用pH7-9.5磷酸盐缓冲液溶解,称取透明质酸加纯化水溶解滴入上述溶液中,磁力搅拌2~6h;加还原试剂,控制温度8~20℃反应1~3h,室温反应8~16h;将反应物放入透析袋中,用PBS缓冲溶液透析两天,冷冻干燥冻干,即得透明质酸-NH2;
称取透明质酸-NH2适量,加pH7-9.5 PBS缓冲溶液超声溶解,加聚乳酸-NHS有机溶剂a,室温磁力搅拌反应15-30h,采用有机溶剂a,除去杂质沉淀;用纯化水透析24-72h,除去杂质,冷冻干燥,即得透明质酸-聚乳酸固体;
其中所述有机溶剂a为甲醇、乙醇、乙醚、乙腈、氯仿、二氯甲烷、二甲基亚砜一种或混合物;所述溶剂b为石油醚、冰乙醚、乙酸乙酯的一种或混合物;
所述还原试剂选自氰基硼氢化钠和硼氢化钠。
2.一种抗肿瘤药物自组装胶束组合物,其特征在于:权利要求1中(I)载体与AB、ABA、ABC两亲性聚酯嵌段共聚物,通过自组装新技术,包覆抗肿瘤药物形成多功能复合胶束;多功能复合胶束由亲水性外壳和聚酯环绕的疏水内核组成;
所述透明质酸-聚乳酸与两亲性聚酯共聚物的自组装多功能复合胶束组合物的重量比为1∶0.2~4.0;
一种抗肿瘤药物自组装胶束组合物的制备方法:
(1)称取透明质酸-聚乳酸溶于纯化水,抗肿瘤药物加有机溶剂a超声溶解,将含药相滴入载体水相,置恒温磁力搅拌仪,扩散2~6h得胶束初混悬液;再将上述胶束初混悬液移至透析袋中,在纯化水或磷酸盐缓冲液中透析12~48h,探头超声5~40min,0.22μm微孔滤膜过滤,即得聚合物胶束混悬液;另取聚合物胶束混悬液,加冻干保护剂,分装西林瓶冷冻干燥,即得聚合物胶束固体粉末;或
(2)两亲性聚酯溶解于有机溶剂,抗肿瘤药物溶于有机溶剂;将含药相缓慢滴入透明质酸-聚乳酸载体有机相,置恒温磁力搅拌仪,扩散2-6h,即得聚合物胶束初混悬液;再将上述聚合物胶束初混悬液置旋转蒸发仪挥去有机溶剂,加纯化水分散,即得聚合物胶束混悬液1;另取两亲性聚酯载体加有机溶剂超声溶解,滴入磁力搅拌的聚合物胶束混悬液1中,固体分散自组装2~3h,探头超声5-30min,0.22μm微孔滤膜过滤,即得自组装复合胶束混悬液;另取自组装复合胶束混悬液,加冻干保护剂,分装西林瓶冷冻干燥,即得自组装复合胶束固体粉末;
所述两亲性聚酯为AB、ABA或ABC型两嵌段共聚物、三嵌段共聚物或接枝共聚物,其中两嵌段共聚物选自mPEG-PLA、mPEG-PCL、mPEG-PLGA、mPEG-PAA,三嵌段共聚物选自mPEG-PLLA-PMMD、PLA-PEG-PLA、PCL-PEG-PCL、PLGA-PEG-PLGA或PEG-PAA-PEG;PMMD为聚丙氨酸、聚赖氨酸、聚甘氨酸、聚谷氨酸,m为800~20000;
其中所述有机溶剂为甲醇、乙醇、乙腈、丙酮、乙醚、二氯甲烷、三氯甲烷的一种或两种混合溶剂;
所述聚合物胶束或复合胶束冻干保护剂选自甘露醇、葡萄糖、乳糖、蔗糖或海藻糖中的一种或几种,冻干保护剂用量按载药聚合物胶束混悬液重量计,1份载药聚合物胶束混悬液加入冻干保护剂0.01~0.2份。
3.根据权利要求2所述的抗肿瘤药物自组装胶束组合物,其特征在于所述抗肿瘤药物选自紫杉醇、多烯紫杉醇、阿霉素、冬凌草素、姜黄素、人参皂苷Rh2、柔红霉素、表柔比星、10-羟基喜树碱、9-硝基喜树碱、S-38、甲氨蝶呤、依托泊苷、米托蒽醌或其衍生物、盐酸阿霉素、盐酸拓扑霉素、盐酸柔红霉素;其中抗肿瘤药物与(I)的重量比为1∶1.0~200。
4.根据权利要求2所述的抗肿瘤药物自组装胶束组合物,其特征在于该胶束组合物可用于治疗非小细胞肺癌、乳腺癌、卵巢癌、子宫颈癌、胃癌、肝癌、白血病、肾癌的单一或联合用药。
Priority Applications (1)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
CN201410244165.3A CN104056275B (zh) | 2014-05-30 | 2014-05-30 | 多功能主动靶向透明质酸‑聚乳酸载体合成及其抗肿瘤药物胶束制备方法 |
Applications Claiming Priority (1)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
CN201410244165.3A CN104056275B (zh) | 2014-05-30 | 2014-05-30 | 多功能主动靶向透明质酸‑聚乳酸载体合成及其抗肿瘤药物胶束制备方法 |
Publications (2)
Publication Number | Publication Date |
---|---|
CN104056275A CN104056275A (zh) | 2014-09-24 |
CN104056275B true CN104056275B (zh) | 2017-09-19 |
Family
ID=51544390
Family Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
CN201410244165.3A Expired - Fee Related CN104056275B (zh) | 2014-05-30 | 2014-05-30 | 多功能主动靶向透明质酸‑聚乳酸载体合成及其抗肿瘤药物胶束制备方法 |
Country Status (1)
Country | Link |
---|---|
CN (1) | CN104056275B (zh) |
Families Citing this family (14)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN106943603B (zh) * | 2017-01-24 | 2020-07-14 | 中山大学 | 一种以pH敏感胶束为模板的纳米金壳的制备方法 |
CN107029235A (zh) * | 2017-03-31 | 2017-08-11 | 武汉理工大学 | 多功能协同主动靶向给药系统及其制备和应用 |
CN107141492A (zh) * | 2017-06-08 | 2017-09-08 | 华南理工大学 | 一种具有靶向功能仿病毒结构高分子囊泡及其制备与应用 |
CN109771663B (zh) * | 2017-11-10 | 2022-03-18 | 中国科学院宁波材料技术与工程研究所 | 一种酸响应性抗癌纳米药物的制备及应用 |
CN108926567A (zh) * | 2018-08-17 | 2018-12-04 | 西南大学 | 基于丝素蛋白用于乳腺癌靶向联合化疗的纳米药物及制备 |
CN110078901A (zh) * | 2019-05-24 | 2019-08-02 | 上海典范医疗科技有限公司 | 一种外消旋聚乳酸制备方法 |
CN112791226A (zh) * | 2019-11-14 | 2021-05-14 | 美国发现集团有限公司 | 一种具有抗肿瘤功能的纳米机器人及其制备方法 |
CN111407729A (zh) * | 2020-04-24 | 2020-07-14 | 东南大学 | 布洛芬聚合物前药与阿霉素共组装纳米粒子及其制备方法 |
CN111632153B (zh) * | 2020-06-23 | 2023-02-24 | 宁夏医科大学 | 一种化学基因药物共载的靶向纳米递药系统及其制备方法 |
WO2022045823A1 (ko) * | 2020-08-31 | 2022-03-03 | 주식회사 바임 | 생분해성 고분자 분산체, 이를 포함하는 조성물 및 피부 개선용 시스템 |
CN112675314B (zh) * | 2020-12-31 | 2023-02-24 | 中国药科大学 | 一种骨靶向纳米胶束递送系统及其制备方法 |
CN113712897B (zh) * | 2021-07-23 | 2023-04-25 | 上海中医药大学 | 一种可溶性微针介导的载紫草素的透皮递药系统及其制备 |
CN114409887B (zh) * | 2021-12-15 | 2023-09-01 | 浙江大学杭州国际科创中心 | 一种氟化端基两亲性聚合物、其制备方法及应用 |
WO2023221082A1 (zh) * | 2022-05-20 | 2023-11-23 | 爱美客技术发展股份有限公司 | 一种接枝聚合物材料及其制备方法与应用 |
Citations (1)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN103751795A (zh) * | 2013-05-20 | 2014-04-30 | 中国药科大学 | 透明质酸-抗肿瘤药偶联物及复合纳米粒组合物的制备和应用 |
-
2014
- 2014-05-30 CN CN201410244165.3A patent/CN104056275B/zh not_active Expired - Fee Related
Patent Citations (1)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN103751795A (zh) * | 2013-05-20 | 2014-04-30 | 中国药科大学 | 透明质酸-抗肿瘤药偶联物及复合纳米粒组合物的制备和应用 |
Non-Patent Citations (1)
Title |
---|
《self assembled amphiphilic hyaluronic acid graft copolymers for targeted release of antitumoral drug》;Pitarresi G. et al.;《Journal of Drug Targeting》;20100531;第18卷(第4期);摘要 * |
Also Published As
Publication number | Publication date |
---|---|
CN104056275A (zh) | 2014-09-24 |
Similar Documents
Publication | Publication Date | Title |
---|---|---|
CN104056275B (zh) | 多功能主动靶向透明质酸‑聚乳酸载体合成及其抗肿瘤药物胶束制备方法 | |
Zheng et al. | Improving breast cancer therapy using doxorubicin loaded solid lipid nanoparticles: Synthesis of a novel arginine-glycine-aspartic tripeptide conjugated, pH sensitive lipid and evaluation of the nanomedicine in vitro and in vivo | |
Huang et al. | Biodegradable self-assembled nanoparticles of poly (D, L-lactide-co-glycolide)/hyaluronic acid block copolymers for target delivery of docetaxel to breast cancer | |
Gothwal et al. | Polymeric micelles: recent advancements in the delivery of anticancer drugs | |
Mezghrani et al. | Hepatocellular carcinoma dually-targeted nanoparticles for reduction triggered intracellular delivery of doxorubicin | |
CN101791411B (zh) | 两亲性多糖偶联物及其药物组合物的制备和应用 | |
Shahriari et al. | Synthesis of hyaluronic acid-based polymersomes for doxorubicin delivery to metastatic breast cancer | |
Tang et al. | Paclitaxel prodrug based mixed micelles for tumor-targeted chemotherapy | |
CN101254309A (zh) | 叶酸受体介导靶向乙酰普鲁兰多糖纳米粒及制备方法 | |
CN109010846A (zh) | 聚乙二醇-壳聚糖-姜黄素聚合物、及其载药纳米粒子和制备方法 | |
Zhu et al. | Enhanced tumor targeting and antitumor efficacy via hydroxycamptothecin-encapsulated folate-modified N-succinyl-N′-octyl chitosan micelles | |
Han et al. | Free paclitaxel-loaded E-selectin binding peptide modified micelle self-assembled from hyaluronic acid-paclitaxel conjugate inhibit breast cancer metastasis in a murine model | |
Luo et al. | pH-responsive stearic acid-O-carboxymethyl chitosan assemblies as carriers delivering small molecular drug for chemotherapy | |
CN103006539A (zh) | 一种聚合物胶束药物组合物及其制备方法 | |
CN104162169A (zh) | 一种药物组合物及其制备方法和用途 | |
CN103720675B (zh) | 一种具有氧化还原敏感的姜黄素前药胶束、胶束单体及其制备方法 | |
Ying et al. | Orally administrable therapeutic nanoparticles for the treatment of colorectal cancer | |
Zhang et al. | Zwitterionic targeting doxorubicin-loaded micelles assembled by amphiphilic dendrimers with enhanced antitumor performance | |
CN106176602B (zh) | 一种靶向作用于胃癌组织的多西紫杉醇壳聚糖纳米胶束及制备方法和应用 | |
Zhang et al. | Biodegradable reduction and pH dual-sensitive polymer micelles based on poly (2-ethyl-2-oxazoline) for efficient delivery of curcumin | |
Pandey et al. | Targeted drug delivery and gene therapy through natural biodegradable nanostructures in pharmaceuticals | |
Zhou et al. | Engineering polyzwitterion with acylsulfonamide-based betaine structure for protonated switch of surface chemistry at tumoral pH and reductive responsive drug release of polymeric micelles | |
CN105153428B (zh) | 一种用于粘液渗透的pH响应性高分子胶束及其制备方法 | |
CN103083682B (zh) | 叶酸修饰的壳聚糖季铵盐-紫杉醇聚合物药物及其制备方法和应用 | |
EP3054928A1 (en) | Self-assembled brush block copolymer-nanoparticles for drug delivery |
Legal Events
Date | Code | Title | Description |
---|---|---|---|
C06 | Publication | ||
PB01 | Publication | ||
C10 | Entry into substantive examination | ||
SE01 | Entry into force of request for substantive examination | ||
GR01 | Patent grant | ||
GR01 | Patent grant | ||
CF01 | Termination of patent right due to non-payment of annual fee |
Granted publication date: 20170919 |