CN109453123A - A kind of Combretastatin analog derivative freeze-dried powder and preparation method thereof - Google Patents
A kind of Combretastatin analog derivative freeze-dried powder and preparation method thereof Download PDFInfo
- Publication number
- CN109453123A CN109453123A CN201811372794.9A CN201811372794A CN109453123A CN 109453123 A CN109453123 A CN 109453123A CN 201811372794 A CN201811372794 A CN 201811372794A CN 109453123 A CN109453123 A CN 109453123A
- Authority
- CN
- China
- Prior art keywords
- freeze
- phosphatide
- combretastatin
- analog derivative
- dried powder
- 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.)
- Granted
Links
- 239000000843 powder Substances 0.000 title claims abstract description 39
- 238000002360 preparation method Methods 0.000 title claims abstract description 19
- 150000004814 combretastatins Chemical class 0.000 title claims description 77
- 238000004108 freeze drying Methods 0.000 claims abstract description 15
- 239000003381 stabilizer Substances 0.000 claims abstract description 9
- 238000000034 method Methods 0.000 claims abstract description 7
- 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 abstract description 6
- 239000008103 glucose Substances 0.000 claims abstract description 6
- 150000003904 phospholipids Chemical class 0.000 claims abstract description 6
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Chemical compound O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 16
- CSCPPACGZOOCGX-UHFFFAOYSA-N Acetone Chemical compound CC(C)=O CSCPPACGZOOCGX-UHFFFAOYSA-N 0.000 claims description 12
- 150000001875 compounds Chemical class 0.000 claims description 12
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 claims description 8
- 239000000243 solution Substances 0.000 claims description 8
- 238000001291 vacuum drying Methods 0.000 claims description 8
- IXYCFFBBFRFQJT-UHFFFAOYSA-N 5-[4-(3,5-dimethoxyphenyl)-1h-imidazol-5-yl]-2-methoxyphenol Chemical class COC1=CC(OC)=CC(C2=C(NC=N2)C=2C=C(O)C(OC)=CC=2)=C1 IXYCFFBBFRFQJT-UHFFFAOYSA-N 0.000 claims description 7
- LGZKGOGODCLQHG-CYBMUJFWSA-N 5-[(2r)-2-hydroxy-2-(3,4,5-trimethoxyphenyl)ethyl]-2-methoxyphenol Chemical group C1=C(O)C(OC)=CC=C1C[C@@H](O)C1=CC(OC)=C(OC)C(OC)=C1 LGZKGOGODCLQHG-CYBMUJFWSA-N 0.000 claims description 6
- HEDRZPFGACZZDS-UHFFFAOYSA-N Chloroform Chemical compound ClC(Cl)Cl HEDRZPFGACZZDS-UHFFFAOYSA-N 0.000 claims description 6
- YMWUJEATGCHHMB-UHFFFAOYSA-N Dichloromethane Chemical compound ClCCl YMWUJEATGCHHMB-UHFFFAOYSA-N 0.000 claims description 6
- 238000006243 chemical reaction Methods 0.000 claims description 6
- 239000008215 water for injection Substances 0.000 claims description 6
- JQWAHKMIYCERGA-UHFFFAOYSA-N (2-nonanoyloxy-3-octadeca-9,12-dienoyloxypropoxy)-[2-(trimethylazaniumyl)ethyl]phosphinate Chemical compound CCCCCCCCC(=O)OC(COP([O-])(=O)CC[N+](C)(C)C)COC(=O)CCCCCCCC=CCC=CCCCCC JQWAHKMIYCERGA-UHFFFAOYSA-N 0.000 claims description 5
- FZWLAAWBMGSTSO-UHFFFAOYSA-N Thiazole Chemical compound C1=CSC=N1 FZWLAAWBMGSTSO-UHFFFAOYSA-N 0.000 claims description 5
- 230000015572 biosynthetic process Effects 0.000 claims description 5
- 235000019441 ethanol Nutrition 0.000 claims description 5
- 239000003960 organic solvent Substances 0.000 claims description 5
- 125000001997 phenyl group Chemical group [H]C1=C([H])C([H])=C(*)C([H])=C1[H] 0.000 claims description 5
- 238000003786 synthesis reaction Methods 0.000 claims description 5
- 125000000339 4-pyridyl group Chemical group N1=C([H])C([H])=C([*])C([H])=C1[H] 0.000 claims description 4
- 230000004048 modification Effects 0.000 claims description 4
- 238000012986 modification Methods 0.000 claims description 4
- 235000010482 polyoxyethylene sorbitan monooleate Nutrition 0.000 claims description 4
- 229920000053 polysorbate 80 Polymers 0.000 claims description 4
- 125000005809 3,4,5-trimethoxyphenyl group Chemical group [H]C1=C(OC([H])([H])[H])C(OC([H])([H])[H])=C(OC([H])([H])[H])C([H])=C1* 0.000 claims description 3
- -1 3,5- Dimethoxyphenyl Chemical group 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
- 238000005411 Van der Waals force Methods 0.000 claims description 3
- 229910052739 hydrogen Inorganic materials 0.000 claims description 3
- 239000001257 hydrogen Substances 0.000 claims description 3
- 125000002887 hydroxy group Chemical group [H]O* 0.000 claims description 3
- 150000002460 imidazoles Chemical class 0.000 claims description 3
- 239000008101 lactose Substances 0.000 claims description 3
- 239000000594 mannitol Substances 0.000 claims description 3
- 235000010355 mannitol Nutrition 0.000 claims description 3
- 125000002924 primary amino group Chemical group [H]N([H])* 0.000 claims description 3
- 230000009467 reduction Effects 0.000 claims description 3
- 238000006467 substitution reaction Methods 0.000 claims description 3
- 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 2
- LVNGJLRDBYCPGB-LDLOPFEMSA-N (R)-1,2-distearoylphosphatidylethanolamine Chemical compound CCCCCCCCCCCCCCCCCC(=O)OC[C@H](COP([O-])(=O)OCC[NH3+])OC(=O)CCCCCCCCCCCCCCCCC LVNGJLRDBYCPGB-LDLOPFEMSA-N 0.000 claims description 2
- QFMZQPDHXULLKC-UHFFFAOYSA-N 1,2-bis(diphenylphosphino)ethane Chemical compound C=1C=CC=CC=1P(C=1C=CC=CC=1)CCP(C=1C=CC=CC=1)C1=CC=CC=C1 QFMZQPDHXULLKC-UHFFFAOYSA-N 0.000 claims description 2
- KILNVBDSWZSGLL-KXQOOQHDSA-N 1,2-dihexadecanoyl-sn-glycero-3-phosphocholine Chemical compound CCCCCCCCCCCCCCCC(=O)OC[C@H](COP([O-])(=O)OCC[N+](C)(C)C)OC(=O)CCCCCCCCCCCCCCC KILNVBDSWZSGLL-KXQOOQHDSA-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
- HVXBOLULGPECHP-WAYWQWQTSA-N Combretastatin A4 Chemical compound C1=C(O)C(OC)=CC=C1\C=C/C1=CC(OC)=C(OC)C(OC)=C1 HVXBOLULGPECHP-WAYWQWQTSA-N 0.000 claims description 2
- GZDFHIJNHHMENY-UHFFFAOYSA-N Dimethyl dicarbonate Chemical compound COC(=O)OC(=O)OC GZDFHIJNHHMENY-UHFFFAOYSA-N 0.000 claims description 2
- ZCQWOFVYLHDMMC-UHFFFAOYSA-N Oxazole Chemical compound C1=COC=N1 ZCQWOFVYLHDMMC-UHFFFAOYSA-N 0.000 claims description 2
- OAICVXFJPJFONN-UHFFFAOYSA-N Phosphorus Chemical compound [P] OAICVXFJPJFONN-UHFFFAOYSA-N 0.000 claims description 2
- 229920001213 Polysorbate 20 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
- 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 2
- 230000009471 action Effects 0.000 claims description 2
- 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 2
- 150000004945 aromatic hydrocarbons Chemical class 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
- 229960005537 combretastatin A-4 Drugs 0.000 claims description 2
- HVXBOLULGPECHP-UHFFFAOYSA-N combretastatin A4 Natural products C1=C(O)C(OC)=CC=C1C=CC1=CC(OC)=C(OC)C(OC)=C1 HVXBOLULGPECHP-UHFFFAOYSA-N 0.000 claims description 2
- 150000004292 cyclic ethers Chemical class 0.000 claims description 2
- 229910052736 halogen Inorganic materials 0.000 claims description 2
- 150000002367 halogens Chemical class 0.000 claims description 2
- 229910052698 phosphorus Inorganic materials 0.000 claims description 2
- 239000011574 phosphorus Substances 0.000 claims description 2
- 229920002946 poly[2-(methacryloxy)ethyl phosphorylcholine] polymer Polymers 0.000 claims description 2
- 239000000256 polyoxyethylene sorbitan monolaurate Substances 0.000 claims description 2
- 235000010486 polyoxyethylene sorbitan monolaurate Nutrition 0.000 claims description 2
- 239000000244 polyoxyethylene sorbitan monooleate Substances 0.000 claims description 2
- 229940068977 polysorbate 20 Drugs 0.000 claims description 2
- 229940068968 polysorbate 80 Drugs 0.000 claims description 2
- 239000007787 solid Substances 0.000 claims description 2
- 239000005720 sucrose Substances 0.000 claims description 2
- 238000009834 vaporization Methods 0.000 claims description 2
- 230000008016 vaporization Effects 0.000 claims description 2
- 210000004556 brain Anatomy 0.000 claims 1
- JEIPFZHSYJVQDO-UHFFFAOYSA-N iron(III) oxide Inorganic materials O=[Fe]O[Fe]=O JEIPFZHSYJVQDO-UHFFFAOYSA-N 0.000 claims 1
- 239000008347 soybean phospholipid Substances 0.000 claims 1
- 150000003557 thiazoles Chemical class 0.000 claims 1
- 230000000694 effects Effects 0.000 abstract description 15
- 238000002347 injection Methods 0.000 abstract description 6
- 239000007924 injection Substances 0.000 abstract description 6
- 238000001990 intravenous administration Methods 0.000 abstract description 3
- 238000010253 intravenous injection Methods 0.000 abstract description 3
- 238000004519 manufacturing process Methods 0.000 abstract description 3
- 231100000331 toxic Toxicity 0.000 abstract description 3
- 230000002588 toxic effect Effects 0.000 abstract description 3
- 206010009944 Colon cancer Diseases 0.000 abstract description 2
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 abstract description 2
- 208000029742 colonic neoplasm Diseases 0.000 abstract description 2
- 201000007270 liver cancer Diseases 0.000 abstract description 2
- 208000014018 liver neoplasm Diseases 0.000 abstract description 2
- 208000002154 non-small cell lung carcinoma Diseases 0.000 abstract description 2
- 238000013268 sustained release Methods 0.000 abstract description 2
- 239000012730 sustained-release form Substances 0.000 abstract description 2
- 231100000419 toxicity Toxicity 0.000 abstract description 2
- 230000001988 toxicity Effects 0.000 abstract description 2
- 208000029729 tumor suppressor gene on chromosome 11 Diseases 0.000 abstract description 2
- 230000002238 attenuated effect Effects 0.000 abstract 1
- 238000000265 homogenisation Methods 0.000 abstract 1
- 238000001802 infusion Methods 0.000 abstract 1
- 239000003223 protective agent Substances 0.000 abstract 1
- 230000008685 targeting Effects 0.000 abstract 1
- 239000003814 drug Substances 0.000 description 20
- 229940079593 drug Drugs 0.000 description 18
- 238000005259 measurement Methods 0.000 description 15
- KBPLFHHGFOOTCA-UHFFFAOYSA-N 1-Octanol Chemical compound CCCCCCCCO KBPLFHHGFOOTCA-UHFFFAOYSA-N 0.000 description 10
- 210000004027 cell Anatomy 0.000 description 10
- IAZDPXIOMUYVGZ-UHFFFAOYSA-N Dimethylsulphoxide Chemical compound CS(C)=O IAZDPXIOMUYVGZ-UHFFFAOYSA-N 0.000 description 9
- 229910019142 PO4 Inorganic materials 0.000 description 9
- NBIIXXVUZAFLBC-UHFFFAOYSA-K phosphate Chemical compound [O-]P([O-])([O-])=O NBIIXXVUZAFLBC-UHFFFAOYSA-K 0.000 description 9
- 239000010452 phosphate Substances 0.000 description 9
- 235000021317 phosphate Nutrition 0.000 description 9
- 238000004090 dissolution Methods 0.000 description 7
- 238000005215 recombination Methods 0.000 description 7
- 230000006798 recombination Effects 0.000 description 7
- UHOVQNZJYSORNB-UHFFFAOYSA-N Benzene Chemical compound C1=CC=CC=C1 UHOVQNZJYSORNB-UHFFFAOYSA-N 0.000 description 6
- OKKJLVBELUTLKV-UHFFFAOYSA-N Methanol Chemical compound OC OKKJLVBELUTLKV-UHFFFAOYSA-N 0.000 description 6
- 230000004700 cellular uptake Effects 0.000 description 5
- 239000002131 composite material Substances 0.000 description 5
- WDOGQTQEKVLZIJ-WAYWQWQTSA-N combretastatin a-4 phosphate Chemical compound C1=C(OP(O)(O)=O)C(OC)=CC=C1\C=C/C1=CC(OC)=C(OC)C(OC)=C1 WDOGQTQEKVLZIJ-WAYWQWQTSA-N 0.000 description 4
- 231100000135 cytotoxicity Toxicity 0.000 description 4
- 230000003013 cytotoxicity Effects 0.000 description 4
- 238000011049 filling Methods 0.000 description 4
- 239000001963 growth medium Substances 0.000 description 4
- 238000003756 stirring Methods 0.000 description 4
- 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 description 3
- REDUQXCPUSNJOL-UHFFFAOYSA-N C(C1=CC=CC=C1)NC(CN(C(C1=CC=C(C=C1)C(C)C)=O)CC1=CC=C(C=C1)C(NO)=O)=O Chemical compound C(C1=CC=CC=C1)NC(CN(C(C1=CC=C(C=C1)C(C)C)=O)CC1=CC=C(C=C1)C(NO)=O)=O REDUQXCPUSNJOL-UHFFFAOYSA-N 0.000 description 3
- 230000008901 benefit Effects 0.000 description 3
- LGZKGOGODCLQHG-UHFFFAOYSA-N combretastatin Natural products C1=C(O)C(OC)=CC=C1CC(O)C1=CC(OC)=C(OC)C(OC)=C1 LGZKGOGODCLQHG-UHFFFAOYSA-N 0.000 description 3
- 229960005527 combretastatin A-4 phosphate Drugs 0.000 description 3
- 230000007547 defect Effects 0.000 description 3
- 239000012738 dissolution medium Substances 0.000 description 3
- 125000002467 phosphate group Chemical group [H]OP(=O)(O[H])O[*] 0.000 description 3
- 229920001983 poloxamer Polymers 0.000 description 3
- 239000000651 prodrug Substances 0.000 description 3
- 229940002612 prodrug Drugs 0.000 description 3
- 239000012679 serum free medium Substances 0.000 description 3
- 238000003860 storage Methods 0.000 description 3
- 239000006228 supernatant Substances 0.000 description 3
- JLPULHDHAOZNQI-ZTIMHPMXSA-N 1-hexadecanoyl-2-(9Z,12Z-octadecadienoyl)-sn-glycero-3-phosphocholine Chemical compound CCCCCCCCCCCCCCCC(=O)OC[C@H](COP([O-])(=O)OCC[N+](C)(C)C)OC(=O)CCCCCCC\C=C/C\C=C/CCCCC JLPULHDHAOZNQI-ZTIMHPMXSA-N 0.000 description 2
- CFWRDBDJAOHXSH-SECBINFHSA-N 2-azaniumylethyl [(2r)-2,3-diacetyloxypropyl] phosphate Chemical compound CC(=O)OC[C@@H](OC(C)=O)COP(O)(=O)OCCN CFWRDBDJAOHXSH-SECBINFHSA-N 0.000 description 2
- RVGRUAULSDPKGF-UHFFFAOYSA-N Poloxamer Chemical compound C1CO1.CC1CO1 RVGRUAULSDPKGF-UHFFFAOYSA-N 0.000 description 2
- 238000009825 accumulation Methods 0.000 description 2
- 239000002585 base Substances 0.000 description 2
- 239000011203 carbon fibre reinforced carbon Substances 0.000 description 2
- 238000013461 design Methods 0.000 description 2
- 230000000857 drug effect Effects 0.000 description 2
- 238000000338 in vitro Methods 0.000 description 2
- 239000002502 liposome Substances 0.000 description 2
- 239000007788 liquid Substances 0.000 description 2
- 238000004895 liquid chromatography mass spectrometry Methods 0.000 description 2
- 239000012528 membrane Substances 0.000 description 2
- 239000002105 nanoparticle Substances 0.000 description 2
- 229960000502 poloxamer Drugs 0.000 description 2
- 238000011160 research Methods 0.000 description 2
- 229940083466 soybean lecithin Drugs 0.000 description 2
- 239000000126 substance Substances 0.000 description 2
- IAKHMKGGTNLKSZ-INIZCTEOSA-N (S)-colchicine Chemical compound C1([C@@H](NC(C)=O)CC2)=CC(=O)C(OC)=CC=C1C1=C2C=C(OC)C(OC)=C1OC IAKHMKGGTNLKSZ-INIZCTEOSA-N 0.000 description 1
- CRUILBNAQILVHZ-UHFFFAOYSA-N 1,2,3-trimethoxybenzene Chemical class COC1=CC=CC(OC)=C1OC CRUILBNAQILVHZ-UHFFFAOYSA-N 0.000 description 1
- 241000723375 Colchicum Species 0.000 description 1
- 102000002322 Egg Proteins Human genes 0.000 description 1
- 108010000912 Egg Proteins Proteins 0.000 description 1
- 235000010469 Glycine max Nutrition 0.000 description 1
- 244000068988 Glycine max Species 0.000 description 1
- 206010028980 Neoplasm Diseases 0.000 description 1
- 241001299681 Salix herbacea Species 0.000 description 1
- 206010044565 Tremor Diseases 0.000 description 1
- 102000004243 Tubulin Human genes 0.000 description 1
- 108090000704 Tubulin Proteins 0.000 description 1
- 238000002835 absorbance Methods 0.000 description 1
- 230000001464 adherent effect Effects 0.000 description 1
- 239000003513 alkali Substances 0.000 description 1
- 230000000118 anti-neoplastic effect Effects 0.000 description 1
- 230000000259 anti-tumor effect Effects 0.000 description 1
- 230000006907 apoptotic process Effects 0.000 description 1
- 239000007864 aqueous solution Substances 0.000 description 1
- 210000004369 blood Anatomy 0.000 description 1
- 239000008280 blood Substances 0.000 description 1
- 239000007853 buffer solution Substances 0.000 description 1
- 230000015556 catabolic process Effects 0.000 description 1
- 230000001413 cellular effect Effects 0.000 description 1
- PJANXHGTPQOBST-QXMHVHEDSA-N cis-stilbene Chemical group C=1C=CC=CC=1/C=C\C1=CC=CC=C1 PJANXHGTPQOBST-QXMHVHEDSA-N 0.000 description 1
- 238000002425 crystallisation Methods 0.000 description 1
- 230000008025 crystallization Effects 0.000 description 1
- 238000002784 cytotoxicity assay Methods 0.000 description 1
- 231100000263 cytotoxicity test Toxicity 0.000 description 1
- 238000006731 degradation reaction Methods 0.000 description 1
- 238000001514 detection method Methods 0.000 description 1
- 238000005538 encapsulation Methods 0.000 description 1
- 238000009472 formulation Methods 0.000 description 1
- 230000036541 health Effects 0.000 description 1
- ZFGMDIBRIDKWMY-PASTXAENSA-N heparin Chemical compound CC(O)=N[C@@H]1[C@@H](O)[C@H](O)[C@@H](COS(O)(=O)=O)O[C@@H]1O[C@@H]1[C@@H](C(O)=O)O[C@@H](O[C@H]2[C@@H]([C@@H](OS(O)(=O)=O)[C@@H](O[C@@H]3[C@@H](OC(O)[C@H](OS(O)(=O)=O)[C@H]3O)C(O)=O)O[C@@H]2O)CS(O)(=O)=O)[C@H](O)[C@H]1O ZFGMDIBRIDKWMY-PASTXAENSA-N 0.000 description 1
- 229920000669 heparin Polymers 0.000 description 1
- 238000010874 in vitro model Methods 0.000 description 1
- 238000001727 in vivo Methods 0.000 description 1
- 238000011534 incubation Methods 0.000 description 1
- 238000009776 industrial production Methods 0.000 description 1
- 230000007794 irritation Effects 0.000 description 1
- 150000002632 lipids Chemical class 0.000 description 1
- 239000000463 material Substances 0.000 description 1
- 230000007246 mechanism Effects 0.000 description 1
- 239000000693 micelle Substances 0.000 description 1
- 239000000203 mixture Substances 0.000 description 1
- 238000000465 moulding Methods 0.000 description 1
- 229930014626 natural product Natural products 0.000 description 1
- 210000004681 ovum Anatomy 0.000 description 1
- 229910052760 oxygen Inorganic materials 0.000 description 1
- 238000005192 partition Methods 0.000 description 1
- 239000000825 pharmaceutical preparation Substances 0.000 description 1
- 230000003285 pharmacodynamic effect Effects 0.000 description 1
- 150000003016 phosphoric acids Chemical class 0.000 description 1
- 230000035479 physiological effects, processes and functions Effects 0.000 description 1
- 239000000047 product Substances 0.000 description 1
- 102000004169 proteins and genes Human genes 0.000 description 1
- 108090000623 proteins and genes Proteins 0.000 description 1
- 238000000197 pyrolysis Methods 0.000 description 1
- 238000011002 quantification Methods 0.000 description 1
- 239000002994 raw material Substances 0.000 description 1
- 230000004044 response Effects 0.000 description 1
- 150000003839 salts Chemical class 0.000 description 1
- 229920006395 saturated elastomer Polymers 0.000 description 1
- 238000007789 sealing Methods 0.000 description 1
- 210000002966 serum Anatomy 0.000 description 1
- 239000007962 solid dispersion Substances 0.000 description 1
- 229910052717 sulfur Inorganic materials 0.000 description 1
- 230000001502 supplementing effect Effects 0.000 description 1
- 230000004083 survival effect Effects 0.000 description 1
- 230000002792 vascular Effects 0.000 description 1
- 210000003462 vein Anatomy 0.000 description 1
- 125000000391 vinyl group Chemical group [H]C([*])=C([H])[H] 0.000 description 1
- 229920002554 vinyl polymer Polymers 0.000 description 1
- 230000010148 water-pollination Effects 0.000 description 1
Classifications
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K9/00—Medicinal preparations characterised by special physical form
- A61K9/14—Particulate form, e.g. powders, Processes for size reducing of pure drugs or the resulting products, Pure drug nanoparticles
- A61K9/19—Particulate form, e.g. powders, Processes for size reducing of pure drugs or the resulting products, Pure drug nanoparticles lyophilised, i.e. freeze-dried, solutions or dispersions
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K31/00—Medicinal preparations containing organic active ingredients
- A61K31/075—Ethers or acetals
- A61K31/085—Ethers or acetals having an ether linkage to aromatic ring nuclear carbon
- A61K31/09—Ethers or acetals having an ether linkage to aromatic ring nuclear carbon having two or more such linkages
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K47/00—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient
- A61K47/06—Organic compounds, e.g. natural or synthetic hydrocarbons, polyolefins, mineral oil, petrolatum or ozokerite
- A61K47/24—Organic compounds, e.g. natural or synthetic hydrocarbons, polyolefins, mineral oil, petrolatum or ozokerite containing atoms other than carbon, hydrogen, oxygen, halogen, nitrogen or sulfur, e.g. cyclomethicone or phospholipids
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K9/00—Medicinal preparations characterised by special physical form
- A61K9/0012—Galenical forms characterised by the site of application
- A61K9/0019—Injectable compositions; Intramuscular, intravenous, arterial, subcutaneous administration; Compositions to be administered through the skin in an invasive manner
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K9/00—Medicinal preparations characterised by special physical form
- A61K9/0012—Galenical forms characterised by the site of application
- A61K9/0053—Mouth and digestive tract, i.e. intraoral and peroral administration
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P35/00—Antineoplastic agents
Landscapes
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Pharmacology & Pharmacy (AREA)
- Veterinary Medicine (AREA)
- Public Health (AREA)
- General Health & Medical Sciences (AREA)
- Animal Behavior & Ethology (AREA)
- Medicinal Chemistry (AREA)
- Epidemiology (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Engineering & Computer Science (AREA)
- General Chemical & Material Sciences (AREA)
- Organic Chemistry (AREA)
- Molecular Biology (AREA)
- Biophysics (AREA)
- Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
- Oil, Petroleum & Natural Gas (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Dermatology (AREA)
- Nutrition Science (AREA)
- Physiology (AREA)
- Medicinal Preparation (AREA)
- Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
Abstract
本发明公开了一种康普瑞汀类衍生物冻干粉针及其制备方法。所述冻干粉针由康普瑞汀衍生物磷脂复合物、稳定剂和冻干保护剂构成，制备方法包括：康普瑞汀衍生物磷脂复合物的制备；高压均质；冷冻干燥。冻干粉针溶解于5％葡萄糖或0.9％生理盐水注射液中使用，可用于静脉注射、滴注或直接口服以治疗非小细胞肺癌、肝癌、结肠癌。本发明制备的冻干粉针药脂比高，毒副作用小，可改善康普瑞汀类衍生物的亲水性和亲脂性，具有一定缓释效果和被动靶向，能提高口服生物利用度，降低静脉给药剂量，减毒增效，复溶性良好，且该制备方法工艺过程简单，易于工业化生产。
The invention discloses a freeze-dried powder injection of compretin derivatives and a preparation method thereof. The freeze-dried powder injection is composed of a compretin derivative phospholipid complex, a stabilizer and a freeze-drying protective agent. The preparation method includes: preparation of the compretin derivative phospholipid complex; high-pressure homogenization; and freeze-drying. The freeze-dried powder for injection is dissolved in 5% glucose or 0.9% normal saline injection, and can be used for intravenous injection, drip infusion or direct oral administration to treat non-small cell lung cancer, liver cancer and colon cancer. The freeze-dried powder for injection prepared by the invention has high drug-to-lipid ratio, small toxic and side effects, can improve the hydrophilicity and lipophilicity of compretin derivatives, has certain sustained-release effect and passive targeting, and can improve oral bioavailability. The intravenous administration dose is reduced, the toxicity is attenuated and the effect is enhanced, and the redissolving property is good, and the preparation method has a simple technological process and is easy to industrialize production.
Description
Technical field
The present invention relates to pharmaceutical preparation and its preparation technical fields, and in particular to a kind of Combretastatin analog derivative freeze-dried powder
Needle and preparation method thereof.
Background technique
Combretastatin (Combretastatin) is a kind of cis- hexichol separated from the bark of African shrub dwarf willow tree
Vinyl natural products, wherein Combretastatin A4 (hereinafter referred to as CA4) be in all structures cytotoxicity it is most strong,
A kind of simplest compound of structure, the benzene that 3,4, the 5- trimethoxies with the connection of cis- carbon-carbon double bond linking arm replace
The basic structure of ring and 3- hydroxyl -4- methoxy substitution phenyl ring.The antitumor mechanism of CA4 predominantly inhibits tubulin poly-
It closes, induce cell apoptosis and antineoplastic vascular is acted on, action target spot is similar with colchicin, and activity is substantially better than colchicum
Alkali.
But CA4 water solubility and fat-soluble difference, and its cis-stilbene structure-activity is stronger but unstable, therefore at present
It is substantially carried out the structural modification of 3 aspects: 1. modifying A ring structure；2. modifying B ring structure；3. modifying carbon-carbon double bond bridging bond
Structure.The stronger CA4 derivative of activity is obtained by structural modification, effective dose is low, and toxic side effect is smaller when use, has huge
Potential applicability in clinical practice.The CA4 derivative, mother nucleus structure are as follows:
Wherein: A N, O or S；R3, which preferably is selected from, can form Van der Waals force, hydrogen bond with phosphatide with phenyl, hydroxyl, amino etc.
The effects of structure.Wherein according to patent CN201110422678, preferably active structure is structure 1:4- (3,5- dimethoxy)-
5- (4- anisyl) oxazole, structure 2:2- phenyl -4- (3,4,5- trimethoxyphenyl) -5- (4- pyridyl group) thiazole, structure
3:4- (3,5- dimethoxy phenyl) -5- (3- hydroxyl -4- methoxyphenyl) imidazoles.
But the water solubility of CA4 derivative is still very poor, limits their clinical application, therefore it is water-soluble to design and prepare its
Property derivative prodrug is current research hotspot.So far the research of the CA4 derivative of domestic and foreign literature report is confined to design more
With its phosphate prodrug of synthesis.OxiGene company, the U.S. has designed and synthesized its phosphate prodrug CA4 phosphate
(combretastatin A4 phosphate, CA4P), comes into the clinical and experimental study stage at present.Although CA4P is improved
The water solubility of CA4, but it cannot stablize storage in aqueous solution, therefore need to be prepared as freeze-dried powder storage, freeze-dried powder
Have the defects that the following aspects: 1. CA4P freeze-dried powder is also easy to produce degradation after redissolving, and stability is poor；2. phos-phate forms liposoluble
Property it is poor, be not easily accessible cell and play drug effect, effective dose is high；3. internal toxic side effect is big, degrade after intravenous administration
Quick-release is eliminated fastly, is not had slow release effect；4. being confined to intravenous administration, do not have the convenience and compliance of oral administration.
In addition, though the method that drug is directly prepared into the new formulations such as liposome, solid dispersion nanoparticle, polymeric micelle can
Improve drug itself, but that there are drugloading rates is low, the problems such as encapsulation rate is unstable, corresponding freeze-dried powder there are Similar Problems,
And preparation process, supplementary product consumption, auxiliary material safety and storage etc. also can Shortcomings.
In existing patent, Combretastatin analog derivative phosphatide complexes related invention is had no.
Summary of the invention
It is an object of the invention to the drawbacks described above to solve Combretastatin analog derivative freeze-dried powder, provide a kind of health
Puri spit of fland analog derivative phosphatide complexes.
A further object of the present invention is to provide a kind of Combretastatin analog derivative phosphatide complexes freeze-dried powder and its system
Preparation Method.
The purpose of the present invention can be achieved through the following technical solutions:
A kind of Combretastatin analog derivative phosphatide complexes are made of Combretastatin analog derivative with phosphatide, Combretastatin class
The molar ratio of derivative and phosphatide is 2:1~1:10, preferably 1:3.
Wherein Combretastatin analog derivative is imidazoles, oxazole and the thiazole of 3,5- Dimethoxyphenyl of the A ring containing 4 substitutions
Class CombretastatinA4 derivative, preferably has phenyl, hydroxyl, and amino etc. can form Van der Waals force, hydrogen bond etc. with phosphatide
The structure of effect, preferably 4- (3,5- dimethoxy) -5- (4- anisyl) oxazole, 2- phenyl -4- (3,4,5- trimethoxy-benzenes
Base) -5- (4- pyridyl group) thiazole, 4- (3,5- dimethoxy phenyl) -5- (3- hydroxyl -4- methoxyphenyl) imidazoles, it is further excellent
Select 4- (3,5- dimethoxy phenyl) -5- (3- hydroxyl -4- methoxyphenyl) imidazoles.
Wherein phosphatide is selected from natural phosphatide and synthesis source phosphatide；The preferred soybean ovum of natural phosphatide
Phosphatide, egg yolk lecithin, cephalin, sphingomyelin, serinephosphatide；The preferred hydrogenated phospholipid of the synthesis source phosphatide,
The analog of PMPC, DPPC, DMPC, DPPE, DSPE, DPPSA and its structural modification.
Combretastatin analog derivative phosphatide complexes of the present invention are to prepare Combretastatin analog derivative phosphatide compound
Application in object freeze-dried powder.
A kind of Combretastatin analog derivative phosphatide complexes freeze-dried powder is answered by the Combretastatin analog derivative phosphatide
Object, stabilizer and freeze drying protectant is closed to constitute.
Wherein stabilizer is selected from PLURONICS F87, Polysorbate 80, Polysorbate 20 or phosphatide and any combination thereof.
Wherein freeze drying protectant is selected from mannitol, lactose, glucose, trehalose, sorbierite or sucrose and any combination thereof.
A kind of preparation method of the Combretastatin derivative freeze-dried powder, comprising the following steps:
(1) Combretastatin analog derivative and phosphatide are dissolved in organic solvent (a), control reaction density is 0.5~20mg/
ML, and stand at a temperature of 0~100 DEG C and compound to be greater than 0.5h；
(2) compound organic solvent (a) used is removed using reduction vaporization and vacuum drying method, obtains Combretastatin
Analog derivative phosphatide complexes solid；
(3) Combretastatin analog derivative phosphatide complexes are distributed to the water for injection dissolved with stabilizer and freeze drying protectant
In, it is high-pressure homogeneous, cross film, freeze-drying；Wherein, Solutions in Freeze-drying each component weight percent is Combretastatin class phosphatide complexes
0.1%~2.0%, stabilizer 0~10%, freeze drying protectant 2~10%.
The reaction dissolvent (a) is preferably selected from aromatic hydrocarbons, halogen derivatives, cyclic ethers, methylene chloride, chloroform, acetone, second
One of alcohol is a variety of, preferably ethyl alcohol.
Wherein reaction density is preferably 3mg/mL, and reaction temperature is preferably 25 DEG C, and recombination time is preferably 12h.
The Combretastatin analog derivative phosphatide complexes freeze-dried powder of method preparation can be redissolved in 5% glucose or 0.9%
In normal saline solution, partial size is preferably 180.7 ± 23.9nm after redissolution, and PDI is 0.108 ± 0.021.
Combretastatin analog derivative phosphatide complexes freeze-dried powder of the present invention is dissolved in 5% glucose or 0.9% physiology salt
It is used in water injection, can be used for being injected intravenously, instils or directly take orally to treat non-small cell lung cancer, liver cancer, colon cancer.
Combretastatin analog derivative phosphatide complexes freeze-dried powder provided by the invention, has the advantage that
1, preparation process is simple, and introducing organic solvent toxicity is low, is suitable for industrial production；
2, compared with liposome, identical molal quantity can load more CA4 derivatives, and drugloading rate is high, reduce phosphatide and use
Amount；
3, the slip for forming and acting on reduction drug of phospholipid molecule and specific CA4 derivative molecular；
4, structure of phospholipid has amphipathic, compared with CA4 derivative, can significantly improve CA4 derivative phosphatide composite
Hydrophily and lipophilicity, lipid membrane can improve intake of the cell to drug；
5, CA4 derivative phosphatide composite is administered orally, and compared with CA4 derivative phosphate, oral bio benefit can be improved
Expenditure；
6, CA4 derivative phosphatide composite intravenous injection administration, compared with CA4 derivative phosphate, prepares and stored
Stability is good in journey, there is certain slow release effect and pH to respond, and nanoparticle has certain passive target effect, can reduce
Dosage improves drug effect, the defect for overcoming CA4 derivative phosphate dosage high；
7, CA4 derivative phosphatide composite can be substantially reduced that CA4 derivative is oral and the intravenous injection of CA4 derivative phosphate
Irritation.
Detailed description of the invention
Fig. 1: the release in vitro measurement of Combretastatin analog derivative phosphatide complexes freeze-dried powder.
Fig. 2: Combretastatin analog derivative phosphatide complexes freeze-dried powder cellular uptake measurement.
Fig. 3: Combretastatin analog derivative phosphatide complexes freeze-dried powder cytotoxicity assay.
Fig. 4: Combretastatin analog derivative phosphatide complexes oral administration biaavailability measurement.
Specific embodiment
In order to make the objectives, technical solutions, and advantages of the present invention clearer, with reference to embodiments, to the present invention
Further details of explanation.It should be appreciated that particular embodiments described herein is not used to limit this hair to illustrate the present invention
Bright range.
Embodiment 1
Precision weighs molal weight ratio 2:1 Combretastatin analog derivative (4- (3,5- dimethoxy) -5- (4- anisyl)
Oxazole) and soybean lecithin in 50mL round-bottomed flask, 5mL acetone is added, control drug concentration is 20mg/mL, is stirred to complete
After fully dissolved, after compound 1h is stood at 30 DEG C, vacuum drying obtains Combretastatin analog derivative phosphatide complexes, measures recombination rate
It is 50%.
Embodiment 2
Precision weighs molal weight ratio 2:1 Combretastatin analog derivative (2- phenyl -4- (3,4,5- trimethoxyphenyl) -
5- (4- pyridyl group) thiazole) and soybean lecithin in 50mL round-bottomed flask, be added 200mL acetone, control drug concentration be
0.5mg/mL is stirred to after being completely dissolved, and after compound 2h is stood at 35 DEG C, vacuum drying obtains Combretastatin analog derivative phosphatide
Compound, measurement recombination rate are 72%
Embodiment 3
Precision weighs molal weight ratio 1:1 Combretastatin analog derivative (4- (3,5- dimethoxy phenyl) -5- (3- hydroxyl -4-
Methoxyphenyl) imidazoles) and egg yolk lecithin in 50mL round-bottomed flask, control drug concentration be 3mg/mL, be added 30mL benzene,
Stirring is to after being completely dissolved, and after compound 2h is stood at 40 DEG C, vacuum drying obtains Combretastatin analog derivative phosphatide complexes, surveys
Determining recombination rate is 62%.
Embodiment 4
Precision weighs molal weight ratio 1:2 Combretastatin analog derivative (4- (3,5- dimethoxy phenyl) -5- (3- hydroxyl -4-
Methoxyphenyl) imidazoles) and egg yolk lecithin in 50mL round-bottomed flask, control drug concentration be 2mg/mL, be added 40mL second
Alcohol is stirred to after being completely dissolved, and after compound 2h is stood at 50 DEG C, it is compound that vacuum drying obtains Combretastatin analog derivative phosphatide
Object, measurement recombination rate are 62%.
Embodiment 5
Precision weighs molal weight ratio 1:4 Combretastatin analog derivative (4- (3,5- dimethoxy phenyl) -5- (3- hydroxyl -4-
Methoxyphenyl) imidazoles) and cephalin in 50mL round-bottomed flask, be added 35mL chloroform, control drug concentration be 2.5mg/mL,
Stirring is to after being completely dissolved, and after compound 12h is stood at 20 DEG C, vacuum drying obtains Combretastatin analog derivative phosphatide complexes,
Measuring recombination rate is 53%.
Embodiment 6
Precision weighs molal weight ratio 1:3 Combretastatin analog derivative (4- (3,5- dimethoxy phenyl) -5- (3- hydroxyl -4-
Methoxyphenyl) imidazoles) and egg yolk lecithin in 50mL round-bottomed flask, be added 30mL ethyl alcohol, control drug concentration be 3mg/
ML, stirring make it completely dissolved, and after compound 12h is stood at 25 DEG C, it is compound that vacuum drying obtains Combretastatin analog derivative phosphatide
Object, measurement recombination rate are 97%.
Embodiment 7
Weigh 188 500mg of Combretastatin analog derivative phosphatide complexes (embodiment 1) 100mg, Poloxamer
(5%), glucose 200mg (2%), dissolution are scattered in 10mL water for injection, filling in 10mL cillin bottle after high-pressure homogeneous,
Freeze-drying, freeze-dried powder appearance collapse, and partial size is 242.1 ± 36.9nm after measurement is redissolved.
Embodiment 8
Weigh Combretastatin analog derivative phosphatide complexes (embodiment 2) 100mg, Tween-80 200mg (2%), lactose
500mg (5%), dissolution are scattered in 10mL water for injection, filling in 10mL cillin bottle after high-pressure homogeneous, are freeze-dried, and are frozen
Dry powder appearance is more fluffy, and partial size is 221.6 ± 45.6nm after measurement is redissolved.
Embodiment 9
Weigh 188 300mg of Combretastatin analog derivative phosphatide complexes (embodiment 6) 100mg, Poloxamer
(3%), mannitol 500mg (5%), dissolution are scattered in 10mL water for injection, filling in 10mL cillin bottle after high-pressure homogeneous,
Freeze-drying, the molding of freeze-dried powder appearance, partial size is 180.7 ± 23.9nm after measurement is redissolved.
Embodiment 10
The measurement of solubility and apparent partition coefficients
It is measured using fask oscillating method (GB/T 21853-2008), measures the pre-saturated water 20mL of n-octyl alcohol respectively and set ground three
In the bottle of angle, each 3 parts, it is separately added into excessive Combretastatin analog derivative 4- (3,5- dimethoxy phenyl) -5- (3- hydroxyl -4- first
Phenyl) Combretastatin analog derivative phosphatide complexes described in imidazoles and embodiment 6,25 DEG C of shaking 12h to dissolution equilibrium move
Into centrifuge tube, 2000 × g is centrifuged 20min.Upper liquid 10mL is taken, is respectively set in ground triangular flask, n-octyl alcohol 10mL is separately added into,
In 25 DEG C of shakings to dissolution equilibrium, 2000 × g is centrifuged 20min.Water phase is pipetted respectively, n-octyl alcohol phase 2mL is set in 10mL measuring bottle, use
Methanol constant volume measures Combretastatin analog derivative and Combretastatin analog derivative phosphatide complexes in one water system of n-octyl alcohol
Solubility.It is shown in Table 1.The results show that solubility improves about 8 times in Combretastatin analog derivative phosphatide complexes water, LogP is improved
About 1.2 times.
The solubility of 1 Combretastatin analog derivative of table and its phosphatide complexes in one water system of n-octyl alcohol
Embodiment 11
Release in vitro measurement.
It takes 1mL Combretastatin analog derivative phosphatide complexes freeze-dried powder (embodiment 6) to redissolve object respectively and is placed in bag filter
It is interior, meet sink conditions, both ends sealing clamp opening is respectively put into 200mL dissolution medium, and dissolution medium is containing 1%
The PBS buffer solution of pH7.4, pH6.6 and pH5.5 of Tween80 investigate its release conditions under 37 DEG C, 100rmp stirring.
C118P group respectively at 5min, 10min, 15min, 0.5h, 1h, 2h, 4h, 6h, 8h, 12h, respectively at 0.5h, 1h, 2h, 4h, 6h,
8h, 12h take 2mL dissolution medium, while supplementing isometric 37 DEG C of fresh dissolution mediums, and sample crosses 0.22 μm of filter membrane, LC-MS
Sample introduction records peak area, calculates the Accumulation dissolution at each time point, draws release profiles.See Fig. 1.The results show that Combretastatin
Analog derivative phosphatide complexes freeze-dried powder preparation group release in vitro model meets drug release under the conditions of pH 7.4 and 6.6 pH
First order kinetics, zero order kinetics are met under the conditions of pH 5.5, and drug release has obvious slow release effect and pH response existing
As it is contemplated that the drug release of preparation in vivo has sustained release and tumor tissues accumulation ability.
Embodiment 12
The measurement of cell pharmacodynamics
Cellular uptake: setting blank, Combretastatin analog derivative, Combretastatin analog derivative phosphate, Combretastatin class
Four groups of derivative phosphatide composite freeze-dried powder (preparation of embodiment 9).Take the A549 cell of logarithm production period with 5 × 106A/hole
It is inoculated in 6 orifice plates, is cultivated for 24 hours in 37 DEG C of cell incubators of complete culture solution, keep its adherent, discard original culture medium, with not
Culture solution containing serum is incubated for 15min, and every hole is added 1500 μ L (the 0.0375 μ L containing DMSO) and is diluted to serum free medium
The serum free medium of 1500 μ L (the 0.0375 μ L containing DMSO) not drug containing is added in 25nM medicine effective concentration, setting control group,
It is incubated in 37 DEG C, investigates uptake ratio and absorbs the relationship of time.Be incubated for different time (1h, 2h, 3h), observation cellular morphology and
Density quickly discards culture medium, and 4 DEG C of PBS are added and terminate cellular uptake, and rinse cell 3 times.200 μ L cell pyrolysis liquids are added
30min smudge cells are shaken in ice bath, 12000 × g is centrifuged 15min, draws 20 μ L supernatants, and BCA protein quantification takes 50 μ L
200 μ L of methanol is added in supernatant, and vortex 3min, 12000 × g are centrifuged 5min, takes supernatant 100uL, and LC-MS measurement drug contains
It measures (all group n=3).See Fig. 2.The results show that Combretastatin analog derivative phosphatide complexes freeze-dried powder group cellular uptake
Amount be higher than bulk pharmaceutical chemicals and phosphate group, and have conspicuousness, illustrate Combretastatin analog derivative phosphatide complexes freeze-dried powder by
It is easier to be utilized by cellular uptake in its lipophilic characteristics.
Cytotoxicity: the A549 cell of logarithm production period is taken, with 1 × 105A/hole is inoculated in 96 orifice plates, is cultivated completely
37 DEG C of base cultures for 24 hours, remove culture medium, and the correspondence group (medicine of the 100 diluted various concentrations of μ L serum free medium is added in every hole
Object concentration is 2.5,5,10,25,50,100nM), after 37 DEG C are incubated for for 24 hours, the MTT solution of the 5mg/mL of 20 μ L is added in every hole.
After 37 DEG C of incubation 4h, culture medium is discarded, the DMSO of 100 μ L is added, shakes 10min to dissolve the crystallization of first a ceremonial jade-ladle, used in libation, using microplate reader
With measurement absorbance at 570nm, cell survival rate is calculated.Combretastatin analog derivative phosphatide complexes cytotoxicity is strong.See figure
3.The results show that Combretastatin analog derivative phosphatide complexes freeze-dried powder IC50Raw material is below under each concentration of 48h for 24 hours
Medicine and phosphate group, and have conspicuousness, illustrate that the cytotoxicity of Combretastatin analog derivative phosphatide complexes freeze-dried powder is strong
In Combretastatin analog derivative phosphate freeze-dried powder, Combretastatin analog derivative curative effect of medication can be improved, reduce Combretastatin
The clinical administration dosage of analog derivative.
Embodiment 13
Oral administration biaavailability measurement
Rat 18 are chosen, Combretastatin derivative group, common mixt group, Combretastatin analog derivative phosphorus are randomly divided into
Fat complexes group (preparation of embodiment 6).After fasting 12h, with the dosage stomach-filling of 50mg/kg, 0.25 after administration, 0.5,1,1.5,2,
3,4,6,8,12,24,36h eyeground vein clump adopts whole blood 0.3mL, is placed in sodium heparin tubes, 10000rpm is centrifuged 10min, takes
The concentration of upper plasma detection Combretastatin analog derivative.See Fig. 4.The results show that Combretastatin analog derivative phosphatide complexes
Group oral administration biaavailability compared with bulk pharmaceutical chemicals group improves about 7 times, and about 5 times of Increased Plasma Half-life, oral administration overcomes Kang Purui
Spit of fland analog derivative phosphoric acid salt drug is only limited to the defect of intravenously administrable.
Claims (9)
1. a kind of Combretastatin analog derivative phosphatide complexes, which is characterized in that it is made of Combretastatin analog derivative and phosphatide,
Wherein the molar ratio of Combretastatin analog derivative and phosphatide is 2:1~1:10.
2. Combretastatin analog derivative phosphatide complexes according to claim 1, which is characterized in that the Combretastatin class
Derivative is that imidazoles, oxazole and the thiazoles Combretastatin A4 of 3,5- Dimethoxyphenyl of the A ring containing 4 substitutions spread out
Biology, preferably has phenyl, hydroxyl, and amino etc. can form the structure of Van der Waals force, hydrogen bond action with phosphatide；Further preferred 4-
(3,5- dimethoxy) -5- (4- anisyl) oxazole, 2- phenyl -4- (3,4,5- trimethoxyphenyl) -5- (4- pyridyl group)
Thiazole, 4- (3,5- dimethoxy phenyl) -5- (3- hydroxyl -4- methoxyphenyl) imidazoles.
3. Combretastatin analog derivative phosphatide complexes according to claim 1, which is characterized in that the phosphatide is selected from day
The phosphatide and synthesis source phosphatide in right source；Wherein natural phosphatide preferably soya lecithin, egg yolk lecithin, brain phosphorus
Rouge, sphingomyelin, serinephosphatide；The preferred hydrogenated phospholipid of synthesis source phosphatide, PMPC, DPPC, DMPC, DPPE, DSPE,
The analog of DPPSA or its structural modification.
4. Combretastatin analog derivative phosphatide complexes described in claim 1-3 are to prepare Combretastatin analog derivative phosphatide multiple
Close the application in object freeze-dried powder.
5. a kind of Combretastatin analog derivative freeze-dried powder, which is characterized in that by Kang Pu of any of claims 1-3
Auspicious spit of fland analog derivative phosphatide complexes, stabilizer and freeze drying protectant are constituted.
6. Combretastatin analog derivative freeze-dried powder according to claim 5, which is characterized in that the stabilizer is selected from pool
One of Luo Shamu 188, Polysorbate 80, Polysorbate 20 are a variety of.
7. Combretastatin analog derivative freeze-dried powder according to claim 1, which is characterized in that the freeze drying protectant choosing
From one of mannitol, lactose, glucose, trehalose, sorbierite or sucrose or a variety of.
8. the preparation method of Combretastatin derivative freeze-dried powder described in a kind of claim 5, which is characterized in that including following
Step:
(1) Combretastatin analog derivative and phosphatide being dissolved in organic solvent (a), control reaction density is 0.5~20mg/mL,
And it is stood at a temperature of 0~100 DEG C compound greater than 0.5h；
(2) compound organic solvent (a) used is removed using reduction vaporization and vacuum drying method, obtains Combretastatin class and spreads out
Biological phosphatide complexes solid；
(3) Combretastatin analog derivative phosphatide complexes are distributed to dissolved in the water for injection of stabilizer and freeze drying protectant,
It is high-pressure homogeneous, film is crossed, is freeze-dried up to the Combretastatin derivative freeze-dried powder；Wherein Solutions in Freeze-drying each component weight
Percentage is Combretastatin class phosphatide complexes 0.1~2.0%, and stabilizer 0~10%, freeze drying protectant 2~10%, surplus is
Water for injection.
9. preparation method according to claim 6, which is characterized in that the reaction dissolvent (a) is selected from aromatic hydrocarbons, halogen-derived
One of object, cyclic ethers, methylene chloride, chloroform, acetone, ethyl alcohol are a variety of.
Priority Applications (2)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201811372794.9A CN109453123B (en) | 2018-11-19 | 2018-11-19 | A kind of compretin derivatives freeze-dried powder injection and preparation method thereof |
| PCT/CN2018/118108 WO2020103175A1 (en) | 2018-11-19 | 2018-11-29 | Combretastatin derivative lyophilized powder injection and preparation method therefor |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201811372794.9A CN109453123B (en) | 2018-11-19 | 2018-11-19 | A kind of compretin derivatives freeze-dried powder injection and preparation method thereof |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN109453123A true CN109453123A (en) | 2019-03-12 |
| CN109453123B CN109453123B (en) | 2021-05-11 |
Family
ID=65610876
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201811372794.9A Expired - Fee Related CN109453123B (en) | 2018-11-19 | 2018-11-19 | A kind of compretin derivatives freeze-dried powder injection and preparation method thereof |
Country Status (2)
| Country | Link |
|---|---|
| CN (1) | CN109453123B (en) |
| WO (1) | WO2020103175A1 (en) |
Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN110237050A (en) * | 2019-07-11 | 2019-09-17 | 苏州大学 | A kind of compretidine nanoparticle and preparation method thereof |
| CN113024400A (en) * | 2021-03-08 | 2021-06-25 | 沈阳药科大学 | Colchicine derivative and preparation method and application thereof |
| CN114460179A (en) * | 2020-11-09 | 2022-05-10 | 深圳市健翔生物制药有限公司 | Method for measuring in-vitro release degree of degarelix acetate for injection |
Citations (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2006119121A2 (en) * | 2005-04-29 | 2006-11-09 | University Of Louisville Research Foundation, Inc. | Cell-surface decoration with active agents |
| CN102190625A (en) * | 2010-03-18 | 2011-09-21 | 南京圣和药业有限公司 | Stilbene tumor targeted medicine Combretastatin A4 analogs |
| WO2012138694A2 (en) * | 2011-04-07 | 2012-10-11 | Emory University | Compositions comprising saccharide binding moieties and methods for targeted therapy |
| CN102863388A (en) * | 2011-07-05 | 2013-01-09 | 南京圣和药业有限公司 | Tumor targeted drug Combretastatin A4 derivatives |
| CN105288648A (en) * | 2015-10-14 | 2016-02-03 | 东南大学 | Phospholipid compound of hydrophilic drugs as well as pharmaceutical composition and application of phospholipid compound |
| CN105476966A (en) * | 2016-01-29 | 2016-04-13 | 中国药科大学 | Silibinin and phospholipid complex nano freeze-dried powder and preparation method thereof |
| CN106580945A (en) * | 2015-10-14 | 2017-04-26 | 上海天氏利医药科技有限公司 | Combretastatin A4 derivative and preparation thereof |
-
2018
- 2018-11-19 CN CN201811372794.9A patent/CN109453123B/en not_active Expired - Fee Related
- 2018-11-29 WO PCT/CN2018/118108 patent/WO2020103175A1/en active Application Filing
Patent Citations (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2006119121A2 (en) * | 2005-04-29 | 2006-11-09 | University Of Louisville Research Foundation, Inc. | Cell-surface decoration with active agents |
| CN102190625A (en) * | 2010-03-18 | 2011-09-21 | 南京圣和药业有限公司 | Stilbene tumor targeted medicine Combretastatin A4 analogs |
| WO2012138694A2 (en) * | 2011-04-07 | 2012-10-11 | Emory University | Compositions comprising saccharide binding moieties and methods for targeted therapy |
| CN102863388A (en) * | 2011-07-05 | 2013-01-09 | 南京圣和药业有限公司 | Tumor targeted drug Combretastatin A4 derivatives |
| CN105288648A (en) * | 2015-10-14 | 2016-02-03 | 东南大学 | Phospholipid compound of hydrophilic drugs as well as pharmaceutical composition and application of phospholipid compound |
| CN106580945A (en) * | 2015-10-14 | 2017-04-26 | 上海天氏利医药科技有限公司 | Combretastatin A4 derivative and preparation thereof |
| CN105476966A (en) * | 2016-01-29 | 2016-04-13 | 中国药科大学 | Silibinin and phospholipid complex nano freeze-dried powder and preparation method thereof |
Non-Patent Citations (3)
| Title |
|---|
| MICO VICTORIA，等: "Evaluation of lipid-stabilised tripropionin nanodroplets as a delivery route for combretastatin A4", 《INTERNATIONAL JOURNAL OF PHARMACEUTICS》 * |
| 刘汉，等: "康普瑞汀脂质体体外抗乳腺癌拟态血管的作用", 《中国药学杂志》 * |
| 马娟娟，等: "血管抑制剂康普瑞汀和康普瑞汀磷酸二钠纳米制剂研究进展", 《国际药学研究杂志》 * |
Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN110237050A (en) * | 2019-07-11 | 2019-09-17 | 苏州大学 | A kind of compretidine nanoparticle and preparation method thereof |
| CN114460179A (en) * | 2020-11-09 | 2022-05-10 | 深圳市健翔生物制药有限公司 | Method for measuring in-vitro release degree of degarelix acetate for injection |
| CN113024400A (en) * | 2021-03-08 | 2021-06-25 | 沈阳药科大学 | Colchicine derivative and preparation method and application thereof |
Also Published As
| Publication number | Publication date |
|---|---|
| CN109453123B (en) | 2021-05-11 |
| WO2020103175A1 (en) | 2020-05-28 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| Jing et al. | A novel polyethylene glycol mediated lipid nanoemulsion as drug delivery carrier for paclitaxel | |
| CN102580111B (en) | Quercetin hydroxypropyl beta-cyclodextrin clathrate liposome, and preparation method thereof and application thereof | |
| CN107149592A (en) | Biological self-assembly nano-crystalline injection and preparation method with lympha targeted function | |
| CN109453123A (en) | A kind of Combretastatin analog derivative freeze-dried powder and preparation method thereof | |
| Wang et al. | A combined self-assembled drug delivery for effective anti-breast cancer therapy | |
| CN106139151A (en) | Ascorbic acid palmityl ester and the synergistic pharmaceutical composition of antitumor drug | |
| CN103228296B (en) | Bendamustine anionic-atioinic cyclopolysaccharide compositions | |
| CN105999279A (en) | Medicine composition comprising butylphthalide and cosolvent | |
| CN103622909A (en) | Cardiolipin-containing new liposome preparation, and its application in antitumor drugs | |
| CN104826122A (en) | Lipid-modified substance of chlorogenic acid and derivative thereof, preparation method and purification method of the lipid-modified substance | |
| CN100525758C (en) | Garcinolic acid liposome and freezing-drying powdery preparation and its making method | |
| CN104826118A (en) | Application of lipid-modified substance of chlorogenic acid and derivative thereof | |
| CN102526073A (en) | Application of mogrol H9 for preparing antitumor drugs | |
| CN102133184A (en) | Icaritin liposome and preparation method thereof | |
| CN108853056B (en) | Folic acid targeted modification carried doxorubicin hydrochloride and gambogic acid nano-structure lipid carrier preparation and preparation method thereof | |
| CN103585639A (en) | Lactoferrin modified solid lipid nanoparticles, as well as preparation method and application thereof | |
| CN102716089A (en) | Gemcitabine hydrochloride liposome injection | |
| CN105919935B (en) | Sorafenib medicine lipid nano suspension and preparation method thereof | |
| CN104473873B (en) | A kind of Cabazitaxel long circulating liposome injection and preparation method thereof | |
| CN104546722B (en) | Miriplatin lipidosome and preparation method thereof | |
| CN103127060A (en) | Application of chloranthus japonicus alcohol D in preparation of antitumor drugs | |
| Wang et al. | Preparation and anti-tumor activity of PEG-PCL polymersomes loaded with curcumol derivative in HepG2 cell line | |
| CN108434101A (en) | A kind of novel Tivozanib liposomes for anticancer, preparation and its preparation method and application | |
| CN103127061A (en) | Medicine application of chloranthus japonicus alcohol M | |
| CN102397280B (en) | Application of 2 alpha-hydroxy protopanoxadiol medicine |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| GR01 | Patent grant | ||
| GR01 | Patent grant | ||
| CF01 | Termination of patent right due to non-payment of annual fee |
Granted publication date: 20210511 |
|
| CF01 | Termination of patent right due to non-payment of annual fee |