CN102579337B - Long circulation lipid nano-suspension containing docetaxel and preparation method thereof - Google Patents
Long circulation lipid nano-suspension containing docetaxel and preparation method thereof Download PDFInfo
- Publication number
- CN102579337B CN102579337B CN 201210058386 CN201210058386A CN102579337B CN 102579337 B CN102579337 B CN 102579337B CN 201210058386 CN201210058386 CN 201210058386 CN 201210058386 A CN201210058386 A CN 201210058386A CN 102579337 B CN102579337 B CN 102579337B
- Authority
- CN
- China
- Prior art keywords
- docetaxel
- circulation
- long
- suspension
- peg
- 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
- 229960003668 docetaxel Drugs 0.000 title claims abstract description 143
- 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 title claims abstract description 142
- 150000002632 lipids Chemical class 0.000 title claims abstract description 82
- 230000004087 circulation Effects 0.000 title claims abstract description 75
- 238000002360 preparation method Methods 0.000 title claims abstract description 64
- 239000006070 nanosuspension Substances 0.000 title abstract description 56
- 239000000725 suspension Substances 0.000 claims abstract description 62
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims abstract description 32
- 238000000034 method Methods 0.000 claims abstract description 25
- 238000010008 shearing Methods 0.000 claims abstract description 13
- 239000000839 emulsion Substances 0.000 claims description 28
- 239000012153 distilled water Substances 0.000 claims description 24
- OVBPIULPVIDEAO-LBPRGKRZSA-N Folic acid Natural products C=1N=C2NC(N)=NC(=O)C2=NC=1CNC1=CC=C(C(=O)N[C@@H](CCC(O)=O)C(O)=O)C=C1 OVBPIULPVIDEAO-LBPRGKRZSA-N 0.000 claims description 20
- 210000000481 breast Anatomy 0.000 claims description 20
- 239000011724 folic acid Substances 0.000 claims description 16
- 235000019152 folic acid Nutrition 0.000 claims description 14
- 229960000304 folic acid Drugs 0.000 claims description 14
- OVBPIULPVIDEAO-UHFFFAOYSA-N N-Pteroyl-L-glutaminsaeure Natural products C=1N=C2NC(N)=NC(=O)C2=NC=1CNC1=CC=C(C(=O)NC(CCC(O)=O)C(O)=O)C=C1 OVBPIULPVIDEAO-UHFFFAOYSA-N 0.000 claims description 12
- 238000001132 ultrasonic dispersion Methods 0.000 claims description 10
- 125000003929 folic acid group Chemical group 0.000 claims description 6
- JLPULHDHAOZNQI-ZTIMHPMXSA-N 1-hexadecanoyl-2-(9Z,12Z-octadecadienoyl)-sn-glycero-3-phosphocholine Chemical compound CCCCCCCCCCCCCCCC(=O)OC[C@H](COP([O-])(=O)OCC[N+](C)(C)C)OC(=O)CCCCCCC\C=C/C\C=C/CCCCC JLPULHDHAOZNQI-ZTIMHPMXSA-N 0.000 claims description 5
- 230000015572 biosynthetic process Effects 0.000 claims description 4
- 229940083466 soybean lecithin Drugs 0.000 claims description 4
- 206010028980 Neoplasm Diseases 0.000 abstract description 54
- 229920001223 polyethylene glycol Polymers 0.000 abstract description 33
- 230000008685 targeting Effects 0.000 abstract description 30
- 230000000694 effects Effects 0.000 abstract description 13
- 150000003904 phospholipids Chemical class 0.000 abstract description 11
- 231100000331 toxic Toxicity 0.000 abstract description 8
- 230000002588 toxic effect Effects 0.000 abstract description 8
- 210000003022 colostrum Anatomy 0.000 abstract 2
- 235000021277 colostrum Nutrition 0.000 abstract 2
- 238000000265 homogenisation Methods 0.000 abstract 1
- 239000003814 drug Substances 0.000 description 42
- 238000009472 formulation Methods 0.000 description 27
- 229940090044 injection Drugs 0.000 description 27
- 238000002347 injection Methods 0.000 description 27
- 239000007924 injection Substances 0.000 description 27
- 239000000203 mixture Substances 0.000 description 27
- 239000000243 solution Substances 0.000 description 27
- 229940079593 drug Drugs 0.000 description 19
- 239000002105 nanoparticle Substances 0.000 description 18
- 239000002245 particle Substances 0.000 description 16
- 239000002202 Polyethylene glycol Substances 0.000 description 15
- 241000699666 Mus <mouse, genus> Species 0.000 description 13
- 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 description 11
- 229930195725 Mannitol Natural products 0.000 description 11
- 241000699670 Mus sp. Species 0.000 description 11
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 description 11
- 230000005540 biological transmission Effects 0.000 description 11
- 238000001914 filtration Methods 0.000 description 11
- 239000000594 mannitol Substances 0.000 description 11
- 235000010355 mannitol Nutrition 0.000 description 11
- 210000001519 tissue Anatomy 0.000 description 11
- -1 vitamin E succinic acid ester Chemical class 0.000 description 11
- 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 10
- 238000012856 packing Methods 0.000 description 10
- WEVYAHXRMPXWCK-UHFFFAOYSA-N Acetonitrile Chemical compound CC#N WEVYAHXRMPXWCK-UHFFFAOYSA-N 0.000 description 9
- RYGMFSIKBFXOCR-UHFFFAOYSA-N Copper Chemical compound [Cu] RYGMFSIKBFXOCR-UHFFFAOYSA-N 0.000 description 9
- 229910052802 copper Inorganic materials 0.000 description 9
- 239000010949 copper Substances 0.000 description 9
- 238000001035 drying Methods 0.000 description 9
- 239000012982 microporous membrane Substances 0.000 description 9
- IYDGMDWEHDFVQI-UHFFFAOYSA-N phosphoric acid;trioxotungsten Chemical compound O=[W](=O)=O.O=[W](=O)=O.O=[W](=O)=O.O=[W](=O)=O.O=[W](=O)=O.O=[W](=O)=O.O=[W](=O)=O.O=[W](=O)=O.O=[W](=O)=O.O=[W](=O)=O.O=[W](=O)=O.O=[W](=O)=O.OP(O)(O)=O IYDGMDWEHDFVQI-UHFFFAOYSA-N 0.000 description 9
- 238000010186 staining Methods 0.000 description 9
- 239000007864 aqueous solution Substances 0.000 description 7
- 230000008859 change Effects 0.000 description 7
- 238000009826 distribution Methods 0.000 description 7
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 description 6
- OKKJLVBELUTLKV-UHFFFAOYSA-N Methanol Chemical compound OC OKKJLVBELUTLKV-UHFFFAOYSA-N 0.000 description 6
- 238000004108 freeze drying Methods 0.000 description 6
- 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 description 5
- 210000004185 liver Anatomy 0.000 description 5
- 230000008569 process Effects 0.000 description 5
- 230000001225 therapeutic effect Effects 0.000 description 5
- IJGRMHOSHXDMSA-UHFFFAOYSA-N Atomic nitrogen Chemical compound N#N IJGRMHOSHXDMSA-UHFFFAOYSA-N 0.000 description 4
- WQZGKKKJIJFFOK-GASJEMHNSA-N Glucose Chemical compound OC[C@H]1OC(O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-GASJEMHNSA-N 0.000 description 4
- 230000000259 anti-tumor effect Effects 0.000 description 4
- 201000011510 cancer Diseases 0.000 description 4
- 210000004027 cell Anatomy 0.000 description 4
- 238000002474 experimental method Methods 0.000 description 4
- 210000002216 heart Anatomy 0.000 description 4
- 238000000338 in vitro Methods 0.000 description 4
- 238000001727 in vivo Methods 0.000 description 4
- 239000002904 solvent Substances 0.000 description 4
- 210000000952 spleen Anatomy 0.000 description 4
- 239000004094 surface-active agent Substances 0.000 description 4
- 210000003462 vein Anatomy 0.000 description 4
- 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 description 3
- PZNPLUBHRSSFHT-RRHRGVEJSA-N 1-hexadecanoyl-2-octadecanoyl-sn-glycero-3-phosphocholine Chemical compound CCCCCCCCCCCCCCCCCC(=O)O[C@@H](COP([O-])(=O)OCC[N+](C)(C)C)COC(=O)CCCCCCCCCCCCCCC PZNPLUBHRSSFHT-RRHRGVEJSA-N 0.000 description 3
- 239000008280 blood Substances 0.000 description 3
- 210000004369 blood Anatomy 0.000 description 3
- 238000002425 crystallisation Methods 0.000 description 3
- 230000008025 crystallization Effects 0.000 description 3
- 238000005516 engineering process Methods 0.000 description 3
- 239000008103 glucose Substances 0.000 description 3
- 239000000787 lecithin Substances 0.000 description 3
- 235000010445 lecithin Nutrition 0.000 description 3
- 229940067606 lecithin Drugs 0.000 description 3
- 239000003446 ligand Substances 0.000 description 3
- 239000004531 microgranule Substances 0.000 description 3
- 239000012071 phase Substances 0.000 description 3
- 235000010482 polyoxyethylene sorbitan monooleate Nutrition 0.000 description 3
- 229920000053 polysorbate 80 Polymers 0.000 description 3
- 238000011160 research Methods 0.000 description 3
- 238000012216 screening Methods 0.000 description 3
- 235000002639 sodium chloride Nutrition 0.000 description 3
- 239000012086 standard solution Substances 0.000 description 3
- 239000000126 substance Substances 0.000 description 3
- 238000010189 synthetic method Methods 0.000 description 3
- 231100000419 toxicity Toxicity 0.000 description 3
- 230000001988 toxicity Effects 0.000 description 3
- 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 description 2
- FVXDQWZBHIXIEJ-LNDKUQBDSA-N 1,2-di-[(9Z,12Z)-octadecadienoyl]-sn-glycero-3-phosphocholine Chemical compound CCCCC\C=C/C\C=C/CCCCCCCC(=O)OC[C@H](COP([O-])(=O)OCC[N+](C)(C)C)OC(=O)CCCCCCC\C=C/C\C=C/CCCCC FVXDQWZBHIXIEJ-LNDKUQBDSA-N 0.000 description 2
- DSNRWDQKZIEDDB-SQYFZQSCSA-N 1,2-dioleoyl-sn-glycero-3-phospho-(1'-sn-glycerol) Chemical compound CCCCCCCC\C=C/CCCCCCCC(=O)OC[C@H](COP(O)(=O)OC[C@@H](O)CO)OC(=O)CCCCCCC\C=C/CCCCCCCC DSNRWDQKZIEDDB-SQYFZQSCSA-N 0.000 description 2
- SNKAWJBJQDLSFF-NVKMUCNASA-N 1,2-dioleoyl-sn-glycero-3-phosphocholine Chemical compound CCCCCCCC\C=C/CCCCCCCC(=O)OC[C@H](COP([O-])(=O)OCC[N+](C)(C)C)OC(=O)CCCCCCC\C=C/CCCCCCCC SNKAWJBJQDLSFF-NVKMUCNASA-N 0.000 description 2
- NRJAVPSFFCBXDT-HUESYALOSA-N 1,2-distearoyl-sn-glycero-3-phosphocholine Chemical compound CCCCCCCCCCCCCCCCCC(=O)OC[C@H](COP([O-])(=O)OCC[N+](C)(C)C)OC(=O)CCCCCCCCCCCCCCCCC NRJAVPSFFCBXDT-HUESYALOSA-N 0.000 description 2
- RFVFQQWKPSOBED-PSXMRANNSA-N 1-myristoyl-2-palmitoyl-sn-glycero-3-phosphocholine Chemical compound CCCCCCCCCCCCCCCC(=O)O[C@@H](COP([O-])(=O)OCC[N+](C)(C)C)COC(=O)CCCCCCCCCCCCC RFVFQQWKPSOBED-PSXMRANNSA-N 0.000 description 2
- IIZPXYDJLKNOIY-JXPKJXOSSA-N 1-palmitoyl-2-arachidonoyl-sn-glycero-3-phosphocholine Chemical compound CCCCCCCCCCCCCCCC(=O)OC[C@H](COP([O-])(=O)OCC[N+](C)(C)C)OC(=O)CCC\C=C/C\C=C/C\C=C/C\C=C/CCCCC IIZPXYDJLKNOIY-JXPKJXOSSA-N 0.000 description 2
- ANOUKFYBOAKOIR-UHFFFAOYSA-N 3,4-dimethoxyphenylethylamine Chemical compound COC1=CC=C(CCN)C=C1OC ANOUKFYBOAKOIR-UHFFFAOYSA-N 0.000 description 2
- OVMSOCFBDVBLFW-VHLOTGQHSA-N 5beta,20-epoxy-1,7beta,13alpha-trihydroxy-9-oxotax-11-ene-2alpha,4alpha,10beta-triyl 4,10-diacetate 2-benzoate Chemical compound O([C@@H]1[C@@]2(C[C@H](O)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)O)C(=O)C1=CC=CC=C1 OVMSOCFBDVBLFW-VHLOTGQHSA-N 0.000 description 2
- KDCGOANMDULRCW-UHFFFAOYSA-N 7H-purine Chemical compound N1=CNC2=NC=NC2=C1 KDCGOANMDULRCW-UHFFFAOYSA-N 0.000 description 2
- 206010002198 Anaphylactic reaction Diseases 0.000 description 2
- PEDCQBHIVMGVHV-UHFFFAOYSA-N Glycerol Natural products OCC(O)CO PEDCQBHIVMGVHV-UHFFFAOYSA-N 0.000 description 2
- 206010018910 Haemolysis Diseases 0.000 description 2
- XUMBMVFBXHLACL-UHFFFAOYSA-N Melanin Chemical compound O=C1C(=O)C(C2=CNC3=C(C(C(=O)C4=C32)=O)C)=C2C4=CNC2=C1C XUMBMVFBXHLACL-UHFFFAOYSA-N 0.000 description 2
- 241001465754 Metazoa Species 0.000 description 2
- 101001000212 Rattus norvegicus Decorin Proteins 0.000 description 2
- 244000162450 Taxus cuspidata Species 0.000 description 2
- 235000009065 Taxus cuspidata Nutrition 0.000 description 2
- SORGEQQSQGNZFI-UHFFFAOYSA-N [azido(phenoxy)phosphoryl]oxybenzene Chemical compound C=1C=CC=CC=1OP(=O)(N=[N+]=[N-])OC1=CC=CC=C1 SORGEQQSQGNZFI-UHFFFAOYSA-N 0.000 description 2
- 125000002252 acyl group Chemical group 0.000 description 2
- 235000001014 amino acid Nutrition 0.000 description 2
- 208000003455 anaphylaxis Diseases 0.000 description 2
- 239000002246 antineoplastic agent Substances 0.000 description 2
- 229940041181 antineoplastic drug Drugs 0.000 description 2
- 150000001875 compounds Chemical class 0.000 description 2
- 230000034994 death Effects 0.000 description 2
- UREBDLICKHMUKA-CXSFZGCWSA-N dexamethasone Chemical compound C1CC2=CC(=O)C=C[C@]2(C)[C@]2(F)[C@@H]1[C@@H]1C[C@@H](C)[C@@](C(=O)CO)(O)[C@@]1(C)C[C@@H]2O UREBDLICKHMUKA-CXSFZGCWSA-N 0.000 description 2
- 229960003957 dexamethasone Drugs 0.000 description 2
- 238000010586 diagram Methods 0.000 description 2
- FVJZSBGHRPJMMA-UHFFFAOYSA-N distearoyl phosphatidylglycerol Chemical compound CCCCCCCCCCCCCCCCCC(=O)OCC(COP(O)(=O)OCC(O)CO)OC(=O)CCCCCCCCCCCCCCCCC FVJZSBGHRPJMMA-UHFFFAOYSA-N 0.000 description 2
- 239000002552 dosage form Substances 0.000 description 2
- 238000009513 drug distribution Methods 0.000 description 2
- 238000011156 evaluation Methods 0.000 description 2
- 230000008588 hemolysis Effects 0.000 description 2
- 238000004128 high performance liquid chromatography Methods 0.000 description 2
- 230000002401 inhibitory effect Effects 0.000 description 2
- 238000011081 inoculation Methods 0.000 description 2
- 238000001990 intravenous administration Methods 0.000 description 2
- 210000004072 lung Anatomy 0.000 description 2
- 210000002540 macrophage Anatomy 0.000 description 2
- 125000001570 methylene group Chemical group [H]C([H])([*:1])[*:2] 0.000 description 2
- 229910052757 nitrogen Inorganic materials 0.000 description 2
- 238000011275 oncology therapy Methods 0.000 description 2
- 230000003285 pharmacodynamic effect Effects 0.000 description 2
- 229920002946 poly[2-(methacryloxy)ethyl phosphorylcholine] polymer Polymers 0.000 description 2
- 102000005962 receptors Human genes 0.000 description 2
- 108020003175 receptors Proteins 0.000 description 2
- 235000004400 serine Nutrition 0.000 description 2
- 239000011780 sodium chloride Substances 0.000 description 2
- 229960002668 sodium chloride Drugs 0.000 description 2
- 230000001954 sterilising effect Effects 0.000 description 2
- 238000004659 sterilization and disinfection Methods 0.000 description 2
- 239000006228 supernatant Substances 0.000 description 2
- 238000012360 testing method Methods 0.000 description 2
- 238000011282 treatment Methods 0.000 description 2
- 230000005909 tumor killing Effects 0.000 description 2
- 210000001835 viscera Anatomy 0.000 description 2
- 238000005303 weighing Methods 0.000 description 2
- 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 description 1
- KLFKZIQAIPDJCW-GPOMZPHUSA-N 1,2-dihexadecanoyl-sn-glycero-3-phosphoserine Chemical compound CCCCCCCCCCCCCCCC(=O)OC[C@H](COP(O)(=O)OC[C@H](N)C(O)=O)OC(=O)CCCCCCCCCCCCCCC KLFKZIQAIPDJCW-GPOMZPHUSA-N 0.000 description 1
- OWEGMIWEEQEYGQ-UHFFFAOYSA-N 100676-05-9 Natural products OC1C(O)C(O)C(CO)OC1OCC1C(O)C(O)C(O)C(OC2C(OC(O)C(O)C2O)CO)O1 OWEGMIWEEQEYGQ-UHFFFAOYSA-N 0.000 description 1
- UAXAYRSMIDOXCU-BJDJZHNGSA-N 2-[[(2r)-2-[[(2s)-2-[[2-[[(2s)-4-amino-2-[[(2r)-2-amino-3-sulfanylpropanoyl]amino]-4-oxobutanoyl]amino]acetyl]amino]-5-(diaminomethylideneamino)pentanoyl]amino]-3-sulfanylpropanoyl]amino]acetic acid Chemical compound SC[C@H](N)C(=O)N[C@@H](CC(N)=O)C(=O)NCC(=O)N[C@@H](CCCN=C(N)N)C(=O)N[C@@H](CS)C(=O)NCC(O)=O UAXAYRSMIDOXCU-BJDJZHNGSA-N 0.000 description 1
- LRYZPFWEZHSTHD-HEFFAWAOSA-O 2-[[(e,2s,3r)-2-formamido-3-hydroxyoctadec-4-enoxy]-hydroxyphosphoryl]oxyethyl-trimethylazanium Chemical class CCCCCCCCCCCCC\C=C\[C@@H](O)[C@@H](NC=O)COP(O)(=O)OCC[N+](C)(C)C LRYZPFWEZHSTHD-HEFFAWAOSA-O 0.000 description 1
- 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 1
- MSTNYGQPCMXVAQ-KIYNQFGBSA-N 5,6,7,8-tetrahydrofolic acid Chemical compound N1C=2C(=O)NC(N)=NC=2NCC1CNC1=CC=C(C(=O)N[C@@H](CCC(O)=O)C(O)=O)C=C1 MSTNYGQPCMXVAQ-KIYNQFGBSA-N 0.000 description 1
- 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 description 1
- IYMAXBFPHPZYIK-BQBZGAKWSA-N Arg-Gly-Asp Chemical compound NC(N)=NCCC[C@H](N)C(=O)NCC(=O)N[C@@H](CC(O)=O)C(O)=O IYMAXBFPHPZYIK-BQBZGAKWSA-N 0.000 description 1
- 208000031648 Body Weight Changes Diseases 0.000 description 1
- 206010006187 Breast cancer Diseases 0.000 description 1
- 208000026310 Breast neoplasm Diseases 0.000 description 1
- FBPFZTCFMRRESA-FSIIMWSLSA-N D-Glucitol Natural products OC[C@H](O)[C@H](O)[C@@H](O)[C@H](O)CO FBPFZTCFMRRESA-FSIIMWSLSA-N 0.000 description 1
- FBPFZTCFMRRESA-JGWLITMVSA-N D-glucitol Chemical compound OC[C@H](O)[C@@H](O)[C@H](O)[C@H](O)CO FBPFZTCFMRRESA-JGWLITMVSA-N 0.000 description 1
- 229920002307 Dextran Polymers 0.000 description 1
- GZDFHIJNHHMENY-UHFFFAOYSA-N Dimethyl dicarbonate Chemical compound COC(=O)OC(=O)OC GZDFHIJNHHMENY-UHFFFAOYSA-N 0.000 description 1
- 102000002322 Egg Proteins Human genes 0.000 description 1
- 108010000912 Egg Proteins Proteins 0.000 description 1
- 108090000790 Enzymes Proteins 0.000 description 1
- 102000004190 Enzymes Human genes 0.000 description 1
- 229930091371 Fructose Natural products 0.000 description 1
- 239000005715 Fructose Substances 0.000 description 1
- RFSUNEUAIZKAJO-ARQDHWQXSA-N Fructose Chemical compound OC[C@H]1O[C@](O)(CO)[C@@H](O)[C@@H]1O RFSUNEUAIZKAJO-ARQDHWQXSA-N 0.000 description 1
- 241000287828 Gallus gallus Species 0.000 description 1
- ZWZWYGMENQVNFU-UHFFFAOYSA-N Glycerophosphorylserin Natural products OC(=O)C(N)COP(O)(=O)OCC(O)CO ZWZWYGMENQVNFU-UHFFFAOYSA-N 0.000 description 1
- 101001105486 Homo sapiens Proteasome subunit alpha type-7 Proteins 0.000 description 1
- 102000001706 Immunoglobulin Fab Fragments Human genes 0.000 description 1
- 108010054477 Immunoglobulin Fab Fragments Proteins 0.000 description 1
- 108010021625 Immunoglobulin Fragments Proteins 0.000 description 1
- 102000008394 Immunoglobulin Fragments Human genes 0.000 description 1
- 241000581650 Ivesia Species 0.000 description 1
- 238000012449 Kunming mouse Methods 0.000 description 1
- 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 description 1
- GUBGYTABKSRVRQ-PICCSMPSSA-N Maltose Natural products O[C@@H]1[C@@H](O)[C@H](O)[C@@H](CO)O[C@@H]1O[C@@H]1[C@@H](CO)OC(O)[C@H](O)[C@H]1O GUBGYTABKSRVRQ-PICCSMPSSA-N 0.000 description 1
- 108010047562 NGR peptide Proteins 0.000 description 1
- 206010029155 Nephropathy toxic Diseases 0.000 description 1
- 206010033128 Ovarian cancer Diseases 0.000 description 1
- 206010061535 Ovarian neoplasm Diseases 0.000 description 1
- 229910019142 PO4 Inorganic materials 0.000 description 1
- 229930012538 Paclitaxel Natural products 0.000 description 1
- 229920003171 Poly (ethylene oxide) Polymers 0.000 description 1
- 239000004698 Polyethylene Substances 0.000 description 1
- 102100021201 Proteasome subunit alpha type-7 Human genes 0.000 description 1
- CZPWVGJYEJSRLH-UHFFFAOYSA-N Pyrimidine Chemical compound C1=CN=CN=C1 CZPWVGJYEJSRLH-UHFFFAOYSA-N 0.000 description 1
- MTCFGRXMJLQNBG-UHFFFAOYSA-N Serine Natural products OCC(N)C(O)=O MTCFGRXMJLQNBG-UHFFFAOYSA-N 0.000 description 1
- 229930006000 Sucrose Natural products 0.000 description 1
- 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 description 1
- 229940123237 Taxane Drugs 0.000 description 1
- AOBORMOPSGHCAX-UHFFFAOYSA-N Tocophersolan Chemical compound OCCOC(=O)CCC(=O)OC1=C(C)C(C)=C2OC(CCCC(C)CCCC(C)CCCC(C)C)(C)CCC2=C1C AOBORMOPSGHCAX-UHFFFAOYSA-N 0.000 description 1
- 102000002070 Transferrins Human genes 0.000 description 1
- 108010015865 Transferrins Proteins 0.000 description 1
- 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 description 1
- 229930003427 Vitamin E Natural products 0.000 description 1
- 238000010521 absorption reaction Methods 0.000 description 1
- CGIHPACLZJDCBQ-UHFFFAOYSA-N acibenzolar Chemical compound SC(=O)C1=CC=CC2=C1SN=N2 CGIHPACLZJDCBQ-UHFFFAOYSA-N 0.000 description 1
- 230000009471 action Effects 0.000 description 1
- 230000004913 activation Effects 0.000 description 1
- 230000009056 active transport Effects 0.000 description 1
- 230000006978 adaptation Effects 0.000 description 1
- 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 description 1
- 150000001413 amino acids Chemical class 0.000 description 1
- 239000002870 angiogenesis inducing agent Substances 0.000 description 1
- 238000010171 animal model Methods 0.000 description 1
- 108010072041 arginyl-glycyl-aspartic acid Proteins 0.000 description 1
- 229930014667 baccatin III Natural products 0.000 description 1
- 230000008901 benefit Effects 0.000 description 1
- 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 description 1
- GUBGYTABKSRVRQ-QUYVBRFLSA-N beta-maltose Chemical compound OC[C@H]1O[C@H](O[C@H]2[C@H](O)[C@@H](O)[C@H](O)O[C@@H]2CO)[C@H](O)[C@@H](O)[C@@H]1O GUBGYTABKSRVRQ-QUYVBRFLSA-N 0.000 description 1
- 230000017531 blood circulation Effects 0.000 description 1
- 230000037396 body weight Effects 0.000 description 1
- 230000004579 body weight change Effects 0.000 description 1
- 201000008275 breast carcinoma Diseases 0.000 description 1
- 239000004359 castor oil Substances 0.000 description 1
- 235000019438 castor oil Nutrition 0.000 description 1
- 239000003795 chemical substances by application Substances 0.000 description 1
- 238000004140 cleaning Methods 0.000 description 1
- 239000005515 coenzyme Substances 0.000 description 1
- 238000001246 colloidal dispersion Methods 0.000 description 1
- 210000003792 cranial nerve Anatomy 0.000 description 1
- 239000013078 crystal Substances 0.000 description 1
- GVJHHUAWPYXKBD-UHFFFAOYSA-N d-alpha-tocopherol Natural products OC1=C(C)C(C)=C2OC(CCCC(C)CCCC(C)CCCC(C)C)(C)CCC2=C1C GVJHHUAWPYXKBD-UHFFFAOYSA-N 0.000 description 1
- 238000003381 deacetylation reaction Methods 0.000 description 1
- BIABMEZBCHDPBV-UHFFFAOYSA-N dipalmitoyl phosphatidylglycerol Chemical compound CCCCCCCCCCCCCCCC(=O)OCC(COP(O)(=O)OCC(O)CO)OC(=O)CCCCCCCCCCCCCCC BIABMEZBCHDPBV-UHFFFAOYSA-N 0.000 description 1
- 238000007599 discharging Methods 0.000 description 1
- 238000012377 drug delivery Methods 0.000 description 1
- 230000002708 enhancing effect Effects 0.000 description 1
- 230000007613 environmental effect Effects 0.000 description 1
- 150000002148 esters Chemical class 0.000 description 1
- 239000000284 extract Substances 0.000 description 1
- 229940064302 folacin Drugs 0.000 description 1
- 229940014144 folate Drugs 0.000 description 1
- 102000006815 folate receptor Human genes 0.000 description 1
- 108020005243 folate receptor Proteins 0.000 description 1
- WIGCFUFOHFEKBI-UHFFFAOYSA-N gamma-tocopherol Natural products CC(C)CCCC(C)CCCC(C)CCCC1CCC2C(C)C(O)C(C)C(C)C2O1 WIGCFUFOHFEKBI-UHFFFAOYSA-N 0.000 description 1
- 229940093181 glucose injection Drugs 0.000 description 1
- ZEMPKEQAKRGZGQ-XOQCFJPHSA-N glycerol triricinoleate Natural products CCCCCC[C@@H](O)CC=CCCCCCCCC(=O)OC[C@@H](COC(=O)CCCCCCCC=CC[C@@H](O)CCCCCC)OC(=O)CCCCCCCC=CC[C@H](O)CCCCCC ZEMPKEQAKRGZGQ-XOQCFJPHSA-N 0.000 description 1
- 239000008187 granular material Substances 0.000 description 1
- 230000012010 growth Effects 0.000 description 1
- 229920000669 heparin Polymers 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
- 229960001008 heparin sodium Drugs 0.000 description 1
- 230000002440 hepatic effect Effects 0.000 description 1
- 229940099578 hydrogenated soybean lecithin Drugs 0.000 description 1
- 230000007062 hydrolysis Effects 0.000 description 1
- 238000006460 hydrolysis reaction Methods 0.000 description 1
- 230000005847 immunogenicity Effects 0.000 description 1
- 238000010253 intravenous injection Methods 0.000 description 1
- FZWBNHMXJMCXLU-BLAUPYHCSA-N isomaltotriose Chemical compound O[C@@H]1[C@@H](O)[C@H](O)[C@@H](CO)O[C@@H]1OC[C@@H]1[C@@H](O)[C@H](O)[C@@H](O)[C@@H](OC[C@@H](O)[C@@H](O)[C@H](O)[C@@H](O)C=O)O1 FZWBNHMXJMCXLU-BLAUPYHCSA-N 0.000 description 1
- 210000003734 kidney Anatomy 0.000 description 1
- 210000001865 kupffer cell Anatomy 0.000 description 1
- 239000008101 lactose Substances 0.000 description 1
- 239000002502 liposome Substances 0.000 description 1
- 239000007791 liquid phase Substances 0.000 description 1
- 231100000053 low toxicity Toxicity 0.000 description 1
- 238000004519 manufacturing process Methods 0.000 description 1
- 238000013507 mapping Methods 0.000 description 1
- 238000013178 mathematical model Methods 0.000 description 1
- 230000010534 mechanism of action Effects 0.000 description 1
- 230000001404 mediated effect Effects 0.000 description 1
- 201000001441 melanoma Diseases 0.000 description 1
- 238000005374 membrane filtration Methods 0.000 description 1
- 230000004060 metabolic process Effects 0.000 description 1
- 238000002156 mixing Methods 0.000 description 1
- 230000004048 modification Effects 0.000 description 1
- 238000012986 modification Methods 0.000 description 1
- 210000000865 mononuclear phagocyte system Anatomy 0.000 description 1
- 239000002159 nanocrystal Substances 0.000 description 1
- 230000007694 nephrotoxicity Effects 0.000 description 1
- 231100000417 nephrotoxicity Toxicity 0.000 description 1
- 208000002154 non-small cell lung carcinoma Diseases 0.000 description 1
- 239000003921 oil Substances 0.000 description 1
- 210000000056 organ Anatomy 0.000 description 1
- 239000003960 organic solvent Substances 0.000 description 1
- 210000004681 ovum Anatomy 0.000 description 1
- 229960001592 paclitaxel Drugs 0.000 description 1
- 239000000825 pharmaceutical preparation Substances 0.000 description 1
- NBIIXXVUZAFLBC-UHFFFAOYSA-K phosphate Chemical compound [O-]P([O-])([O-])=O NBIIXXVUZAFLBC-UHFFFAOYSA-K 0.000 description 1
- 239000010452 phosphate Substances 0.000 description 1
- 239000008363 phosphate buffer Substances 0.000 description 1
- 230000035790 physiological processes and functions Effects 0.000 description 1
- 229940113116 polyethylene glycol 1000 Drugs 0.000 description 1
- 239000002243 precursor Substances 0.000 description 1
- QEVHRUUCFGRFIF-MDEJGZGSSA-N reserpine Chemical compound O([C@H]1[C@@H]([C@H]([C@H]2C[C@@H]3C4=C(C5=CC=C(OC)C=C5N4)CCN3C[C@H]2C1)C(=O)OC)OC)C(=O)C1=CC(OC)=C(OC)C(OC)=C1 QEVHRUUCFGRFIF-MDEJGZGSSA-N 0.000 description 1
- 238000005070 sampling Methods 0.000 description 1
- 238000004062 sedimentation Methods 0.000 description 1
- 150000003355 serines Chemical class 0.000 description 1
- 239000008354 sodium chloride injection Substances 0.000 description 1
- 239000000600 sorbitol Substances 0.000 description 1
- 238000005728 strengthening Methods 0.000 description 1
- 239000005720 sucrose Substances 0.000 description 1
- 238000003786 synthesis reaction Methods 0.000 description 1
- 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 description 1
- 238000002560 therapeutic procedure Methods 0.000 description 1
- 210000001541 thymus gland Anatomy 0.000 description 1
- 210000004881 tumor cell Anatomy 0.000 description 1
- 208000029729 tumor suppressor gene on chromosome 11 Diseases 0.000 description 1
- 229940088594 vitamin Drugs 0.000 description 1
- 229930003231 vitamin Natural products 0.000 description 1
- 235000013343 vitamin Nutrition 0.000 description 1
- 239000011782 vitamin Substances 0.000 description 1
- 229940046009 vitamin E Drugs 0.000 description 1
- 235000019165 vitamin E Nutrition 0.000 description 1
- 239000011709 vitamin E Substances 0.000 description 1
- 150000003722 vitamin derivatives Chemical class 0.000 description 1
- 239000008215 water for injection Substances 0.000 description 1
Images
Abstract
The invention discloses a long circulation lipid nano-suspension containing docetaxel. The nano-suspension is prepared from the following components in parts by weight: 0.5-50 parts of water solution, 0.5-50 parts of docetaxel, 0.05-20 parts of lipid-PEG (poly(ethylene glycol)) and 50-90 parts of phospholipid or is prepared from 0.5-50 parts of water solution, 0.5-50 parts of docetaxel, 0.05-20 parts of lipid-PEG, 50-90 parts of phospholipid and 0.01-10 parts of targeting factor with tumor targeting function. A preparation method of the nano-suspension comprises the following steps of: (1) dissolving phospholipid and lipid-PEG into the water solution to obtain solution A, or dissolving phospholipid, lipid-PEG and the targeting factor with tumor targeting function into the water solution to obtain solution A; (2) dispersing docetaxel into the solution A obtained in the step (1) to obtain suspension B; (3) carrying out high speed shearing on the suspension B for 1-3 minutes to obtain colostrum C; and (4) homogenizing the colostrum C by a high pressure homogenization method, thus obtaining the long circulation lipid nano-suspension. The preparation has the effects of increasing the content of docetaxel and reducing the toxic or side effect of docetaxel.
Description
Technical field
The present invention relates to contain long-circulation fat matter nano suspension, targeting lipids nano suspension and the lyophilized formulations thereof of docetaxel, with and preparation method thereof, medical technical field belonged to.
Background technology
Docetaxel (docetaxel) is that the precursor compound 10-deacetylation baccatin III that extracts from Taxus baccata (Taxusbaccata L) is through the semi-synthetic taxoids that obtains.The mechanism of action of docetaxel and taxanes seemingly, anti-tumor activity is 1.3～12 times of paclitaxel, and the kinds of tumor such as breast carcinoma, nonsmall-cell lung cancer and ovarian cancer are all had definite curative effect.The docetaxel structural formula is: C
43H
53NO
14, molecular weight is 807.88, structural formula is as follows:
The very low 3 μ g/ml that are about of the dissolubility of docetaxel in water, oral administration biaavailability is poor, less than 8%, for improving its bioavailability, use clinically at present its injection, the listing dosage form is Docetaxel for Injection and docetaxel injection, and such preparation dissolves, dilutes and could use with 5% glucose injection or 0.9% sodium chloride injection before intravenous drip.Also have Tween 80 and ethanol equal solvent in such preparation, easily produce haemolysis and anaphylactic reaction, need take in advance the medical treatments such as dexamethasone, clinical application is inconvenient, and drug safety is low.Place easily oxidized, hydrolysis because docetaxel is long-time in solution, thereby reduce the content of principal agent docetaxel, so the quality stability of injection is poor.So far, there is no solution of the above problems.
Nano suspension is a kind of submicron colloidal dispersion of pure medicament nano granule, take surfactant as solubilizing agent.Compare with lipid system, this technology can be successfully made preparation with the medicine that not only is insoluble in water but also is insoluble in oil, overcome and prepared the shortcoming that other preparation needs first dissolved substance, improve the bioavailability of insoluble drug by simple technique and prescription, avoid simultaneously a large amount of supplementary elements to patient's toxic and side effects.Keep best crystalline state when preparation prepares and had enough little drug particle size.The particulate nature of this dosage form can change the intravenous Pharmacokinetic Characteristics of medicine in addition, and the effect of high-efficiency low-toxicity is arranged; The particle promotion stripping that diminishes can solve many and the low relevant problem of oral administration biaavailability; Medicine is solid-state its chemically stable that makes; The small-particle sedimentation is slow, makes its physically stable.We once applied for a Chinese invention patent (201011486.0), related to a kind of docetaxel nanometer crystallization preparation and lyophilized preparation and preparation method of containing.It is characterized in that the present invention utilizes surfactant as solubilizing agent, through the high pressure breast is even, docetaxel is prepared into nanocrystal solution, obtain the docetaxel nanometer crystal lyophilized preparation finally by filtration sterilization, lyophilization.Studies show that, this preparation can increase the dissolubility of docetaxel, can increase to a certain extent drug level in tumor, improves antitumous effect, and the toxic action of body is obviously reduced.
This drug-supplying system belongs to the passive target drug-supplying system, medicine carrying microgranule is easily absorbed by the macrophage of monokaryon-macrophage system (the especially kupffer cell of liver), be transported to the organs such as liver, spleen by normal physiological processes, the target site that reach other is just had any problem.If to the medicine carrying microgranule surface modifying the picked-up that to avoid macrophage, prevent from concentrating in the liver spleen, changed microgranule in vivo distribution and arrive specific target site and realize initiatively targeting, thereby accumulate and active transport is realized target administration to tumor tissues by passive.
Long circular nanometer suspensoid (long circulation nano-suspensions) claims that also stealthy nano suspension (stealth nano-suspensions) is an important research direction.The long circular nanometer suspensoid that hydrophilic Polyethylene Glycol (PEG) is modified can be escaped catching of reticuloendothelial system, significant prolongation its holdup time in blood circulation, be detained and osmosis (EPR) prolong drug circulation time in vivo by enhancing, increase the accumulating in the tumor tissues by bag medicine carrying thing, thereby improved the targeting that cancer therapy drug transmits.
Long circular nanometer suspensoid drug-supplying system still belongs to the passive target drug-supplying system.For further strengthening the lipid nanometer suspensoid to the selectively targeted ability of tumor tissues, to significantly improving the curative effect of antitumor drug, the research initiatively stealthy nano suspension of targeting becomes the important means that improves the cancer therapy drug targeting.Therefore can modify stealthy nano suspension with folic acid, transferrins and monoclonal antibody etc., to increase the targeting of drug delivery.Folic acid (folic acid, FA) is a kind of micromolecule vitamin, and it can be reduced to tetrahydrofolic acid, and the latter is the coenzyme of one carbon unit transferring enzyme, participates in the de novo synthesis of one carbon unit metabolism and purine, thymus pyrimidine.Compare with other targeted molecular such as monoclonal antibody, the FA relative molecular mass is little, non-immunogenicity, cheap and easy to get, good stability, and medicine or carrier between chemical bonding simple.Therefore build delivery system take FA as targeted molecular and become one of study hotspot of active targeting therapy on tumor.
Summary of the invention
For above-mentioned prior art, the invention provides the long-circulation fat matter nano suspension, targeting lipids nano suspension and the lyophilized formulations thereof that contain docetaxel, and provide its preparation method, preparation of the present invention, improved the content of docetaxel in preparation, reduce the toxic and side effects of docetaxel, effectively increase preparation circulation time in vivo, play long-acting and purpose slow release; In addition, can realize the targeted delivery of medicine, optionally with drug release in tumor locus or its cell interior, improve the drug level of target area, further improve the therapeutic effect of medicine.
The present invention is achieved by the following technical solutions:
A kind of long-circulation fat matter nano suspension that contains docetaxel is to be made by the component that comprises following weight portion: 0.5～50 part of aqueous solution, 0.5～50 part of docetaxel, lipid-PEG0.05～20 part, 0～90 part of phosphatidase 15;
Or: 0.5～50 part of aqueous solution, 0.5～50 part of docetaxel, 0.05～20 part of lipid-PEG, 0～90 part of phosphatidase 15 has 0.01～10 part of the targeting factor of tumor-targeting function;
Prepare by following preparation method:
(1) get phospholipid and lipid-PEG (as, DSPE-PEG, DOPE-PEG, DPPC-PEG), be dissolved in aqueous solution, get solution A;
Or: get phospholipid, lipid-PEG and be connected with the cancer target factor lipid-PEG-ligand (as, DSPE-PEG-FA, DSPE-PEG-NGR, DSPE-PEG-RGD), be dissolved in aqueous solution, get solution A;
(2) docetaxel is scattered in the solution A of step (1) gained, gets suspension B;
(3) with suspension B high speed shear 1～3 minute, get primary emulsion C;
(4) primary emulsion C is adopted the even method of high pressure breast carry out breast even, namely get long-circulation fat matter docetaxel nanometer suspensoid/targeting lipids nano suspension.
Also comprise freeze drying protectant in the described long-circulation fat matter nano suspension that contains docetaxel, the concentration of freeze drying protectant in the lipid nanometer suspensoid is 0.005～0.2mg/ml, and the mode that adds is: add after breast is even.
Preferably, the molecular weight of described Polyethylene Glycol is between 200～5000, preferably between 2000～3000.
described phospholipid is selected from: soybean lecithin (SPC), polyethylene glycol 1000 vitamin E succinic acid ester (TPGS), two myristoyl lecithin (DMPC), DLPC (DLPC), two hard ester acyl lecithin (DPPC), DPPC (DPPC), distearoylphosphatidylcholine (DSPC), MPPC (MPPC), PMPC (PMPC), PSPC (PSPC), SPPC (SPPC), Ovum Gallus domesticus Flavus lecithin (EPC), hydrogenated soybean lecithin (HSPC), DOPC (DOPC), DOPE (DOPE), PE (DLPG), two Petiolus Trachycarpi acyl glycerol (DPPG), DSPG (DSPG), DOPG (DOPG), two myristoyl phosphatidic acid (DMPA), DPPA (DPPA), DMPEA (DMPE), DPPE (DPPE), two myristoyl Phosphatidylserine (DMPS), two palmityl phosphatidyl two serines (DPPS), cephalin acyl serine (PS), cranial nerve sphingomyelins (BSP), two palmityl sphingomyelins (DPSP), distearyl sphingomyelin (DSSP), any in DSPE (DSPE) or several mixing, wherein preferably: soybean lecithin (SPC), DSPE (DSPE) or DOPE (DOPE).
Described targeting factor with tumor-targeting function is selected from: specific angiogenesis factor receptor antibody and Fab fragment thereof, RGD peptide and analog thereof, NGR peptide and analog thereof, folic acid (folic acid, FA), preferred folic acid.
Described aqueous solution is selected from distilled water, 0.9% sodium-chloride water solution, 5% D/W, contain the water for injection of 5% glucose and 0.9% sodium chloride, contain the ringer's inj of 5% glucose or any in phosphate buffer.
In described step (3), the speed of high speed shear is 20000r/min.
In described step (4), the parameter of the even method of high pressure breast is: successively in 200bar circulation 10 times, 500bar circulation 10 times, 1000bar circulation 20 times.This optimised process for finding out through great many of experiments is if circulating pressure is too little or cycle-index can not be made not the crystallization preparation or reach desired particle diameter.
Described freeze drying protectant is selected from: mannitol, sorbitol, sucrose, glucose, lactose, fructose, trehalose, maltose, dextran, aminoacid, amino acid salts, phosphate or Polyethylene Glycol.
The described preparation method that contains the long-circulation fat matter nano suspension of docetaxel, step is as follows:
(1) get phospholipid and lipid-PEG (as, DSPE-PEG, DOPE-PEG, DPPC-PEG), be dissolved in aqueous solution, get solution A;
Or: get phospholipid, lipid-PEG and be connected with the cancer target factor lipid-PEG-ligand (as, DSPE-PEG-FA, DSPE-PEG-NGR, DSPE-PEG-RGD), be dissolved in aqueous solution, get solution A;
(2) docetaxel is scattered in the solution A of step (1) gained, gets suspension B;
(3) with suspension B high speed shear 1～3 minute, get primary emulsion C;
(4) primary emulsion C is adopted the even method of high pressure breast carry out breast even, namely get long-circulation fat matter docetaxel nanometer suspensoid; Or: after breast is even, add freeze drying protectant, filtration sterilization, moisture is removed in lyophilization, gets dry long-circulation fat matter docetaxel nanometer suspensoid.
In the present invention, the synthetic method of lipid-PEG is: phospholipid lipid (as DSPE) is connected with PEG, forms the phospholipid lipid-PEG (DSPE-PEG) that PEG modifies.the synthetic method that is connected with the lipid-PEG-ligand of the cancer target factor is: after obtaining lipid-PEG (DSPE-PEG), the activation of PEG end is formed Acibenzolar lipid-PEG-BTC or lipid-PEG-NHS (DSPE-PEG-BTC or DSPE-PEG-NHS), be connected with the targeting factor with tumor-targeting function (FA as carboxylic in end) again, formation has the targeting factor of tumor-targeting function (as lipid-PEG-F, DSPE-PEG-F) (synthetic method see reference document: Targeting Folate Receptor with Folate Linked to Extremities Poly (ethylene glycol)-Grafted Liposomes:In Vitro Studies).
The present invention has designed a kind of folacin receptor mediated long-circulation fat matter nanometer suspension preparation as novel docetaxel drug-supplying system: the antitumor drug bag is stated from nano suspension, modifies with acquisition long-circulation fat matter nano suspension with hydrophilic Polyethylene Glycol (PEG) on the nano suspension surface; Modify with PEG and the targeting factor FA with tumor-targeting function simultaneously, obtain the targeting lipids nano suspension, its meaning is present in the following aspects: (1) uses the nano suspension bag to carry docetaxel, can effectively reduce the toxic and side effects of docetaxel; (2) carry out the PEGization modification and can effectively increase preparation circulation time in vivo, play long-acting and purpose slow release; (3) end that utilizes modified with folic acid PEG can be realized the targeted delivery of medicine as the targeting Guidance means, optionally with drug release in tumor locus or its cell interior, improve the drug level of target area, improve the bioavailability of medicine.
The long-circulation fat matter nano suspension that contains docetaxel of the present invention, with phospholipid as surfactant, toxicity is low, good biocompatibility, have safe, good stability, the advantage such as toxic and side effects is low, curative effect is reliable, preparation technology is simple, can pass through the membrane filtration degerming, can realize suitability for industrialized production.
The nano suspension that the long-circulation fat matter nano suspension that contains docetaxel of the present invention is modified with PEG is as carrier, bag carrying anti-tumor medicine docetaxel.By modifying with PEG, must improve the long cycle performance of medicine, improve the half-life of medicine, thereby improve the therapeutic effect of tumor.
The nano suspension that the long-circulating target lipid nanometer suspensoid that contains docetaxel of the present invention is modified simultaneously with PEG and tumor locus targeting factor is as carrier, bag carrying anti-tumor medicine docetaxel.By take PEG as bridge, indirect joint tumor locus targeting factor must improve target-oriented drug, also can improve the half-life of medicine simultaneously, thereby improves the therapeutic effect of tumor.
In order to improve the targeting of docetaxel, the half-life of also improving medicine simultaneously, improve the bioavailability of medicine, thereby improve the therapeutic effect of tumor, and reduce the toxic action of medicine, do not add organic solvent or polyoxyethylene castor oil in preparation process of the present invention, and select suitable surfactant and concentration, circulation can form long-circulation fat matter nano suspension, targeting lipids nano suspension and the lyophilized formulations thereof that contains docetaxel for several times at suitable pressures.Due to the strict screening of process conditions of the present invention, resulting nano suspension particle diameter is less, and is not destroyed through the filtering with microporous membrane post crystallization.Get respectively docetaxel long circulation lipid nanometer suspensoid and targeting lipids nano suspension appropriate, drip on copper mesh, carry out negative staining with 2% phosphotungstic acid, after natural drying, (TEM) is lower under transmission electron microscope observes, and particle diameter is between 50～250nm.
Description of drawings
Fig. 1 is the transmission electron microscope photo (* 14000) of nanometer suspension freeze-dried dose of the long-circulation fat matter of embodiment 1 preparation.
Fig. 2 is the transmission electron microscope photo (* 14000) of suspension freeze-dried dose of the long-circulating target lipid nanometer of embodiment 7 preparation.
Fig. 3 is the particle size distribution figure (mean diameter 204.2nm) of nanometer suspension freeze-dried dose of the long-circulation fat matter of embodiment 1 preparation.
Fig. 4 is the particle size distribution figure (mean diameter 220.6nm) of suspension freeze-dried dose of the long-circulating target lipid nanometer of embodiment 7 preparation.
Fig. 5 is the release in vitro curve.
Fig. 6 is the mouse tumor audio-visual picture.
Fig. 7 is the Mouse Weight variation diagram, annotates: * P＜0.05, compare with Docetaxel * * P＜0.01.
Fig. 8 is mouse tumor change in volume figure, annotates:
$P＜0.05,
$ $Compare with the normal saline group P＜0.01, * P＜0.05, and compare with Docetaxel * * P＜0.01,
#P＜0.05,
##Compare with long-circulation fat matter nano suspension P＜0.01.
Fig. 9 is the relation curve (n=5) that after the various preparations of mouse mainline, blood drug level and time change.
Figure 10 is the many handkerchiefs of mouse mainline
Drug level after docetaxel long circulation lipid nanometer suspensoid and docetaxel long-circulating target lipid nanometer suspensoid in each tissue wherein, (a) represents Duo Pa
(b) represent docetaxel long circulation lipid nanometer suspensoid, (c) represent docetaxel long-circulating target lipid nanometer suspensoid.
Figure 11 is the many handkerchiefs of mouse mainline
Drug distribution (n=5) in each time tumor after docetaxel long circulation lipid nanometer suspensoid and docetaxel long-circulating target lipid nanometer suspensoid, wherein, each histogrammic left, center, right represents respectively Duo Pa
Docetaxel long circulation lipid nanometer suspensoid and docetaxel long-circulating target lipid nanometer suspensoid.
The specific embodiment
The present invention is further illustrated below in conjunction with embodiment.
Embodiment 1:
Bag carries the long-circulation fat matter nano suspension of docetaxel
Precision takes SPC: DSPE-PEG (300: 6) (unit: mg), add in the 60ml distilled water dissolving and consist of disperse medium (solution A).Add the 30mg docetaxel, the even suspension B that gets of ultra-sonic dispersion.Continue to adopt high-speed shearing machine 20000r/min high speed shear 1～3min, make primary emulsion C.Then primary emulsion C is adopted the even method of high pressure breast, respectively 200bar circulation 10 times, 500bar circulation 10 times, 1000bar circulation 20 times makes docetaxel long circulation lipid nanometer suspensoid.
The suspension of above preparation adds in the cillin bottle of packing into after 3.0g mannitol, put-80 ℃ of pre-freeze 24h in refrigerator, then put into freezer dryer-40 ℃, 0.5mbar48h, get the suspension freeze-dried preparation of white loose shape docetaxel long circulation lipid nanometer, this lyophilized formulations adds the 2mL distilled water can redissolve fully in 1min through jolting.
Get docetaxel long circulation lipid nanometer suspension appropriate, drip on copper mesh, carry out negative staining with 2% phosphotungstic acid, after natural drying, (TEM) is lower under transmission electron microscope observes, and resulting nano particle diameter is less, and is not destroyed after filtering with microporous membrane.
Make nanoparticle suspension formulations mean diameter 204.2nm.
Before and after lyophilizing, the particle diameter result is as shown in table 1: adopt 3000SH laser particle analyzer (Britain Ma Erwen company), and the change of size before and after three batches of nanoparticle suspension formulations lyophilizing of mensuration preparation, after result shows lyophilizing, particle diameter increases to some extent, has good stability.
The nanometer suspension freeze-dried front and back of table 1. long-circulation fat matter particle diameter result
Embodiment 2:
Blank long-circulation fat matter nano suspension
Precision takes SPC: DSPE-PEG (300: 9) (unit: mg), add in the 60ml distilled water dissolving and consist of disperse medium (solution A).Adopt high-speed shearing machine 20000r/min high speed shear 1～3min, make primary emulsion C.Then primary emulsion C is adopted the even method of high pressure breast, respectively 200bar circulation 10 times, 500bar circulation 10 times, 1000bar circulation 20 times makes blank long-circulation fat matter nano suspension.
The suspension of above preparation adds in the cillin bottle of packing into after 3.0g mannitol, put-80 ℃ of pre-freeze 24h in refrigerator, then put into freezer dryer-40 ℃, 0.5mbar48h, get the nanometer suspension freeze-dried preparation of the blank long-circulation fat matter of white loose shape, this lyophilized formulations adds the 2mL distilled water can redissolve fully in 1min through jolting.
Embodiment 3:
Bag carries the long-circulation fat matter nano suspension of docetaxel
Precision takes SPC: DSPE-PEG (300: 9) (unit: mg), add in the 60ml distilled water dissolving and consist of disperse medium (solution A).Add the 30mg docetaxel, the even suspension B that gets of ultra-sonic dispersion.Continue to adopt high-speed shearing machine 20000r/min high speed shear 1～3min, make primary emulsion C.Then primary emulsion C is adopted the even method of high pressure breast, respectively 200bar circulation 10 times, 500bar circulation 10 times, 1000bar circulation 20 times makes docetaxel long circulation lipid nanometer suspensoid.
The suspension of above preparation adds in the cillin bottle of packing into after 3.0g mannitol, put-80 ℃ of pre-freeze 24h in refrigerator, then put into freezer dryer-40 ℃, 0.5mbar48h, get the suspension freeze-dried preparation of white loose shape docetaxel long circulation lipid nanometer, this lyophilized formulations adds the 2mL distilled water can redissolve fully in 1min through jolting.
Get docetaxel long circulation lipid nanometer suspension appropriate, drip on copper mesh, carry out negative staining with 2% phosphotungstic acid, after natural drying, (TEM) is lower under transmission electron microscope observes, and resulting nano particle diameter is less, and is not destroyed after filtering with microporous membrane.
Make nanoparticle suspension formulations mean diameter 220.1nm.
Embodiment 4:
Bag carries the long-circulation fat matter nano suspension of docetaxel
Precision takes SPC: DSPE-PEG (300: 15) (unit: mg), add in the 60ml distilled water dissolving and consist of disperse medium (solution A).Add the 30mg docetaxel, the even suspension B that gets of ultra-sonic dispersion.Continue to adopt high-speed shearing machine 20000r/min high speed shear 1～3min, make primary emulsion C.Then primary emulsion C is adopted the even method of high pressure breast, respectively 200bar circulation 10 times, 500bar circulation 10 times, 1000bar circulation 20 times makes docetaxel long circulation lipid nanometer suspensoid.
The suspension of above preparation adds in the cillin bottle of packing into after 3.0g mannitol, put-80 ℃ of pre-freeze 24h in refrigerator, then put into freezer dryer-40 ℃, 0.5mbar48h, get the suspension freeze-dried preparation of white loose shape docetaxel long circulation lipid nanometer, this lyophilized formulations adds the 2mL distilled water can redissolve fully in 1min through jolting.
Get docetaxel long circulation lipid nanometer suspension appropriate, drip on copper mesh, carry out negative staining with 2% phosphotungstic acid, after natural drying, (TEM) is lower under transmission electron microscope observes, as shown in Figure 1, due to the strict screening of process conditions of the present invention, resulting nano particle diameter is less, and is not destroyed after filtering with microporous membrane.
Make nanoparticle suspension formulations mean diameter 190.5nm, as shown in Figure 3.
Embodiment 5:
Bag carries the long-circulation fat matter nano suspension of docetaxel
Precision takes SPC: DSPE-PEG (300: 21) (unit: mg), add in the 60ml distilled water dissolving and consist of disperse medium (solution A).Add the 30mg docetaxel, the even suspension B that gets of ultra-sonic dispersion.Continue to adopt high-speed shearing machine 20000r/min high speed shear 1～3min, make primary emulsion C.Then primary emulsion C is adopted the even method of high pressure breast, respectively 200bar circulation 10 times, 500bar circulation 10 times, 1000bar circulation 20 times makes docetaxel long circulation lipid nanometer suspensoid.
The suspension of above preparation adds in the cillin bottle of packing into after 3.0g mannitol, put-80 ℃ of pre-freeze 24h in refrigerator, then put into freezer dryer-40 ℃, 0.5mbar48h, get the suspension freeze-dried preparation of white loose shape docetaxel long circulation lipid nanometer, this lyophilized formulations adds the 2mL distilled water can redissolve fully in 1min through jolting.
Get docetaxel long circulation lipid nanometer suspension appropriate, drip on copper mesh, carry out negative staining with 2% phosphotungstic acid, after natural drying, (TEM) is lower under transmission electron microscope observes, and resulting nano particle diameter is less, and is not destroyed after filtering with microporous membrane.
Make nanoparticle suspension formulations mean diameter 230.3nm.
Embodiment 6:
Bag carries the long-circulation fat matter nano suspension of docetaxel
Precision takes SPC: DSPE-PEG (300: 27) (unit: mg), add in the 60ml distilled water dissolving and consist of disperse medium (solution A).Add the 30mg docetaxel, the even suspension B that gets of ultra-sonic dispersion.Continue to adopt high-speed shearing machine 20000r/min high speed shear 1～3min, make primary emulsion C.Then primary emulsion C is adopted the even method of high pressure breast, respectively 200bar circulation 10 times, 500bar circulation 10 times, 1000bar circulation 20 times makes docetaxel long circulation lipid nanometer suspensoid.
The suspension of above preparation adds in the cillin bottle of packing into after 3.0g mannitol, put-80 ℃ of pre-freeze 24h in refrigerator, then put into freezer dryer-40 ℃, 0.5mbar48h, get the suspension freeze-dried preparation of white loose shape docetaxel long circulation lipid nanometer, this lyophilized formulations adds the 2mL distilled water can redissolve fully in 1min through jolting.
Get docetaxel long circulation lipid nanometer suspension appropriate, drip on copper mesh, carry out negative staining with 2% phosphotungstic acid, after natural drying, (TEM) is lower under transmission electron microscope observes, and resulting nano particle diameter is less, and is not destroyed after filtering with microporous membrane.
Make nanoparticle suspension formulations mean diameter 250.5nm.
Embodiment 7:
Bag carries the long-circulating target lipid nanometer suspensoid of the modified with folic acid of docetaxel
Precision takes SPC: DSPE-PEG: DSPE-PEG-F (300: 5: 1) (unit: mg), add in the 60ml distilled water dissolving and consist of disperse medium (solution A).Add the 30mg docetaxel, the even suspension B that gets of ultra-sonic dispersion.Continue to adopt high-speed shearing machine 20000r/min high speed shear 1～3min, make primary emulsion C.Then primary emulsion C is adopted the even method of high pressure breast, respectively 200bar circulation 10 times, 500bar circulation 10 times, 1000bar circulation 20 times makes docetaxel long-circulating target lipid nanometer suspensoid.
The suspension of above preparation adds in the cillin bottle of packing into after 3.0g mannitol, put-80 ℃ of pre-freeze 24h in refrigerator, then put into freezer dryer-40 ℃, 0.5mbar48h, get the suspension freeze-dried preparation of white loose shape docetaxel long-circulating target lipid nanometer, this lyophilized formulations adds the 2mL distilled water can redissolve fully in 1min through jolting.
Get docetaxel long-circulating target nanosuspension appropriate, drip on copper mesh, carry out negative staining with 2% phosphotungstic acid, after natural drying, (TEM) is lower under transmission electron microscope observes, and resulting nano particle diameter is less, and is not destroyed after filtering with microporous membrane.
Make nanoparticle suspension formulations mean diameter 220.6nm.
Before and after lyophilizing, the particle diameter result is as shown in table 2: adopt 3000SH laser particle analyzer (Britain Ma Erwen company), and the change of size before and after three batches of nanoparticle suspension formulations lyophilizing of mensuration preparation, after result shows lyophilizing, particle diameter increases to some extent, has good stability.
The suspension freeze-dried front and back of table 2. long-circulating target lipid nanometer particle diameter result
Embodiment 8:
Blank long-circulating target lipid nanometer suspensoid
Precision takes SPC: DSPE-PEG: DSPE-PEG-F (300: 5: 1) (unit: mg), add in the 60ml distilled water dissolving and consist of disperse medium (solution A).Adopt high-speed shearing machine 20000r/min high speed shear 1～3min, make primary emulsion C.Then primary emulsion C is adopted the even method of high pressure breast, respectively 200bar circulation 10 times, 500bar circulation 10 times, 1000bar circulation 20 times makes blank long-circulating target lipid nanometer suspensoid.
The suspension of above preparation adds in the cillin bottle of packing into after 3.0g mannitol, put-80 ℃ of pre-freeze 24h in refrigerator, then put into freezer dryer-40 ℃, 0.5mbar48h, get the suspension freeze-dried preparation of the blank long-circulating target lipid nanometer of white loose shape, this lyophilized formulations adds the 2mL distilled water can redissolve fully in 1min through jolting.
Embodiment 9:
Bag carries the long-circulating target lipid nanometer suspensoid of the modified with folic acid of docetaxel
Precision takes SPC: DSPE-PEG: DSPE-PEG-F (300: 15: 6) (unit: mg), add in the 60ml distilled water dissolving and consist of disperse medium (solution A).Add the 30mg docetaxel, the even suspension B that gets of ultra-sonic dispersion.Continue to adopt high-speed shearing machine 20000r/min high speed shear 1～3min, make primary emulsion C.Then primary emulsion C is adopted the even method of high pressure breast, respectively 200bar circulation 10 times, 500bar circulation 10 times, 1000bar circulation 20 times makes docetaxel long-circulating target lipid nanometer suspensoid.
The suspension of above preparation adds in the cillin bottle of packing into after 3.0g mannitol, put-80 ℃ of pre-freeze 24h in refrigerator, then put into freezer dryer-40 ℃, 0.5mbar48h, get the suspension freeze-dried preparation of white loose shape docetaxel long-circulating target lipid nanometer, this lyophilized formulations adds the 2mL distilled water can redissolve fully in 1min through jolting.
Get docetaxel long-circulating target nanosuspension appropriate, drip on copper mesh, carry out negative staining with 2% phosphotungstic acid, after natural drying, (TEM) is lower under transmission electron microscope observes, as shown in Figure 2, due to the strict screening of process conditions of the present invention, resulting nano particle diameter is less, and is not destroyed after filtering with microporous membrane.
Make nanoparticle suspension formulations mean diameter 200.5nm, as shown in Figure 4.
Embodiment 10:
Bag carries the long-circulating target lipid nanometer suspensoid of the modified with folic acid of docetaxel
Precision takes SPC: DSPE-PEG: DSPE-PEG-F (300: 15: 9) (unit: mg), add in the 60ml distilled water dissolving and consist of disperse medium (solution A).Add the 30mg docetaxel, the even suspension B that gets of ultra-sonic dispersion.Continue to adopt high-speed shearing machine 20000r/min high speed shear 1～3min, make primary emulsion C.Then primary emulsion C is adopted the even method of high pressure breast, respectively 200bar circulation 10 times, 500bar circulation 10 times, 1000bar circulation 20 times makes docetaxel long-circulating target lipid nanometer suspensoid.
The suspension of above preparation adds in the cillin bottle of packing into after 3.0g mannitol, put-80 ℃ of pre-freeze 24h in refrigerator, then put into freezer dryer-40 ℃, 0.5mbar48h, get the suspension freeze-dried preparation of white loose shape docetaxel long-circulating target lipid nanometer, this lyophilized formulations adds the 2mL distilled water can redissolve fully in 1min through jolting.
Get docetaxel long-circulating target nanosuspension appropriate, drip on copper mesh, carry out negative staining with 2% phosphotungstic acid, after natural drying, (TEM) is lower under transmission electron microscope observes, and resulting nano particle diameter is less, and is not destroyed after filtering with microporous membrane.
Make nanoparticle suspension formulations mean diameter 230.8nm.
Embodiment 11: the foundation of docetaxel HPLC chromatographic condition and standard curve:
Hypersil ODS post (4.6mm * 250mm, 5 μ m)
Mobile phase: acetonitrile-distilled water (55: 45, V/V);
Sample size: 20 μ L;
Flow velocity: 1.0mL/min;
Detect wavelength: 230nm;
Column temperature: room temperature;
Theoretical cam curve is not less than 6000.
The preparation of standard solution
Precision takes docetaxel standard substance 10mg, with the dissolving of 10ml acetonitrile, obtains the standard solution of 1mg/ml, puts 4 ℃ of Refrigerator stores standby.
The foundation of docetaxel solution standard curve:
Get 7 10ml volumetric flasks, compound concentration is the standard solution of 5,10,20,30,40,50 μ g/ml, by high-efficient liquid phase technique, to concentration (C) mapping, obtains regression equation y=12684x-722.76, r=0.9999 with peak area (A).
Embodiment 12: the release in vitro of the suspension freeze-dried preparation of docetaxel nanometer:
Get respectively embodiment 1, docetaxel long circulation lipid nanometer suspensoid, long-circulating target lipid nanometer suspensoid and commercially available docetaxel injection (many handkerchiefs of embodiment 7 preparations
) solution 0.25mL (C=1.0mg/ml) adds in the pretreated bag filter that shut off value is relative molecular mass 8-12kDa, wherein Docetaxel injection group is matched group, each parallel 3 parts, tighten bag mouth, put into the 15ml release medium (PBS that contains 0.5% Tween 80 is housed, pH=7.4) in EP pipe, then (temperature is 37 ± 0.5 ℃ to be placed in constant temperature oscillator, rotating speed is 100r/min) carry out release test, respectively at 1,2,4,8,12, the 24h sampling, each 1m L release medium of taking out, and add simultaneously 37 ℃ of fresh release medium with volume.Medicament contg by in HPLC method mensuration release medium calculates total release percentage (%), draws release profiles, as shown in Figure 5.
Can find out that by discharging result long-circulation fat matter nano suspension, long-circulating target lipid nanometer suspensoid and Docetaxel injection release in vitro all meet the first order kinetics mathematical model, long-circulation fat matter nano suspension discharge fit equation be ln (100-Q)=-0.084t+4.588, regression coefficient r=0.9985; Long-circulating target lipid nanometer suspensoid discharge fit equation be ln (100-Q)=-0.066t+4.567, regression coefficient r=0.9980; Injection Docetaxel group discharge fit equation be ln (100-Q)=-0.1859t+4.4038, regression coefficient r=0.9991.
Attention: * * P＜0.01, compare with the Docetaxel group
Embodiment 13: pharmacodynamic experiment
40 of the female Kunming mouses of laboratory animal employing 18-22g size, after environmental adaptation in 7 days, Mice Inoculated melanin tumour b16 tumor cell.At first use trypsinization in PBS (pH 7.4) cancerous cell, in right side of mice oxter inoculation, every approximately inoculation 5 * 10
4Individual mouse melanoma B16 cell.When tumor size is about 100mm
3When (approximately 10 days), mice is divided into six groups, every group 10, be respectively normal saline group, blank long-circulation fat matter nano suspension group, blank long-circulating target lipid nanometer suspensoid group, injection Docetaxel group, long-circulation fat matter nano suspension group (embodiment 1 preparation), long-circulating target lipid nanometer suspension formulation group (implementing 7 preparations).Wherein injection Docetaxel group, long-circulation fat matter nano suspension group, long-circulating target lipid nanometer suspensoid group are respectively by tail intravenously administrable 20mg/kg, normal saline group, blank long-circulation fat matter nano suspension group, blank long-circulating target lipid nanometer suspensoid group injection equal-volume solution, be administered once in every three days, three weeks of successive administration.
In experimentation, every other day Mouse Weight is carried out weighing, tumor size is measured.Put to death mice after three weeks, get tumor, weigh, calculate tumour inhibiting rate, draw the Mouse Weight variation diagram, tumor change in volume figure.Result such as following table 3 and Fig. 6,7,8:
Tumour inhibiting rate (%)=(the average tumor of the average tumor weight-test group of matched group is heavy)/average tumor of matched group heavy * 100%
Table 3
Annotate:
$P＜0.05,
$ $Compare with the normal saline group P＜0.01, * P＜0.05, and compare with Docetaxel * * P＜0.01,
#P＜0.05,
##Compare with long-circulation fat matter nano suspension P＜0.01.
Conclusion: in pharmacodynamic experiment, by the mouse tumor audio-visual picture, the mouse tumor multigraph, tumor change in volume figure can find out intuitively that long-circulation fat matter nano suspension group compares with Docetaxel solution group and have obvious tumor killing effect, long-circulating target lipid nanometer suspensoid group has again better therapeutic effect, illustrates that pharmaceutical preparation of the present invention has the drug action of good treatment tumor disease aspect.Docetaxel long circulation lipid nanometer suspensoid group and injection Docetaxel group are relatively, the tumor volume reduces obviously (P＜0.01), long-circulating target lipid nanometer suspensoid group and long-circulation fat matter nano suspension group are relatively, the tumor volume also reduces obviously (P＜0.05), shows that the preparation group compares better efficacy with Docetaxel injection group.Yet, the Mouse Weight variation differs obviously (p＜0.01), normal saline group, blank long-circulation fat matter nano suspension group, blank long-circulating target lipid nanometer suspensoid group, long-circulation fat matter nano suspension group, long-circulating target lipid nanometer suspensoid group Mice Body weight average have growth, and gain percentage is respectively 56.89%, 44.40%, 48.64%, 18.89%, 22.07%.Yet, have injection Docetaxel group Mouse Weight only and descend, drop to 20.95%.Be a key factor of judgement preparation toxicity due to body weight change, therefore can say that the toxic action of docetaxel nanometer suspension formulation is much smaller than commercially available injection Docetaxel.
By above result as can be known, relatively tumor killing effect is better for docetaxel long circulation lipid nanometer suspensoid group, long-circulating target lipid nanometer suspensoid group preparation group and injection Docetaxel group, but the former is significantly less than the latter to body toxicity, and need not to take in advance haemolysis and anaphylactic reaction that the medicine such as dexamethasone brings with Tween 80 and ethanol equal solvent in the control injection in the medication process, thereby improved the convenience of clinical application and safety etc.
Embodiment 14: pharmacokinetics and tissue distribution experiment
Get and inoculate kind of a tumor mice, the tumor volume is 100mm approximately
3, be divided at random 3 groups, 3 of the parallel settings of each time point.The tail vein injection administration, dosage 60mg/kg (all with the docetaxel cubage), fasting 12-18h before administration, freely drink water, first group is injection Docetaxel group, second group is docetaxel long circular nanometer suspension formulation group (embodiment 1 preparation), and the 3rd group is docetaxel targeted nano suspension formulation group (implementing 7 preparations).Get blood respectively at 5min, 15min, 30min, 45min, 1,2,4,6,8,10,12h after (0h), administration before administration in the eyeground vein clump, be placed in the centrifuge tube through the heparin sodium rinse, the centrifugal 10min of 4000r/min gets upper plasma, and-20 ℃ of Refrigerator stores are to be measured.Put to death mice after getting blood, after winning tumor, the heart, liver, spleen, lung, kidney device and being placed in normal saline and cleaning, be stored in-20 ℃ of refrigerators to be measured.
Plasma sample is processed: accurate absorption mice plasma 0.2mL to be measured, be placed in the 1.5mL centrifuge tube, add methanol and acetonitrile solution (50: 50, V/V) 0.5mL is after vortex 1min, the centrifugal 10min of 10000r/min, supernatant is moved in centrifuge tube, 40 ℃, dry up under-50 ℃ of water-bath nitrogen current, be dissolved in 0.1mL mobile phase, get 20 μ L sample introductions.
Tissue sample is processed: get the tested animal internal organs, precise weighing adds 1mL normal saline homogenate (hepatic tissue adds 2mL).Get homogenate sample 0.2mL, put in the 1.5mL centrifuge tube, add acetonitrile and methanol solution (50: 50, V/V) 0.5mL is after vortex 1min, the centrifugal 10min of 10000r/min, supernatant is moved in centrifuge tube, 40 ℃, dry up under-50 ℃ of water-bath nitrogen current, be dissolved in 0.1mL mobile phase, get 20 μ L sample introductions.
1, interior medicine dynamics result of study: as shown in Figure 9.
Calculating such as the table 4 of pharmacokinetic parameter:
Table 4
Annotate: * P＜0.05, compare with Docetaxel * * P＜0.01.
2, mice docetaxel lipid nanometer suspensoid distribution research
As shown in Figure 10,11; By Figure 10 and Figure 11 as can be known, the many handkerchiefs of mouse tail vein injection
After injection, docetaxel is respectively organizing internal organs that distribution is all arranged, and distributes in the majority at the heart and nephridial tissue; After tail vein injection docetaxel long circulation lipid nanometer suspensoid and docetaxel long-circulating target lipid nanometer suspendible, medicine distributes in liver, spleen and lung tissue and increases, and has reduced the distribution of docetaxel in the heart, nephridial tissue, and heart nephrotoxicity reduces.With many handkerchiefs
The injection group is compared, and in lipid nanometer suspensoid group tumor tissue, drug distribution obviously improves, and has obvious tumor-targeting.After administration 8h, the interior drug level of docetaxel long circulation lipid nanometer suspensoid group tumor is 3.19 times of injection group, and the interior drug level of docetaxel long-circulating target lipid nanometer suspendible tumor is 3.48 times of injection group.
3, Evaluation on Its Targeting Performance in Mice Body: as shown in table 5.
The many handkerchiefs of table 5. intravenous injection
Evaluation on Its Targeting Performance (n=5) in docetaxel long circulation lipid nanometer suspensoid and docetaxel long-circulating target lipid nanometer suspensoid Mice Body
These three kinds of docetaxel injectas, total targeting efficient T of docetaxel long circulation lipid nanometer suspensoid group tumor
eBy many handkerchiefs
5.22% of injection group is brought up to 5.67% (1.09 times), its relative uptake ratio r
eWith peak concentration C in tumor
eBe respectively 5.22 and 1.02; Total targeting efficient T of docetaxel long-circulating target lipid nanometer suspensoid group tumor
eBy many handkerchiefs
Injection group 5.22% is brought up to 6.40% (1.23 times), its relative uptake ratio r
eWith peak concentration C in tumor
eBe respectively 5.92 and 1.17.Compare with docetaxel long circulation lipid nanometer suspensoid, docetaxel long-circulating target lipid nanometer suspensoid has better cancer target efficient.
Claims (2)
1. a bag carries the long-circulating target lipid nanometer suspensoid of the modified with folic acid of docetaxel, it is characterized in that: prepare by the following method: precision takes soybean lecithin: PEG-DSPE: PEG-DSPE-folic acid, three's quality is 300:5:1, unit: mg adds to dissolving formation disperse medium in the 60ml distilled water; Add the 30mg docetaxel, the even suspension B that gets of ultra-sonic dispersion; Continue to adopt high-speed shearing machine 20000r/min high speed shear 1～3min, make primary emulsion C; Then primary emulsion C is adopted the even method of high pressure breast, respectively 200bar circulation 10 times, 500bar circulation 10 times, 1000bar circulation 20 times makes docetaxel long-circulating target lipid nanometer suspensoid.
2. a bag carries the preparation method of long-circulating target lipid nanometer suspensoid of the modified with folic acid of docetaxel, it is characterized in that: step is as follows: precision takes soybean lecithin: PEG-DSPE: PEG-DSPE-folic acid, three's quality is 300:5:1, unit: mg adds to dissolving formation disperse medium in the 60ml distilled water; Add the 30mg docetaxel, the even suspension B that gets of ultra-sonic dispersion; Continue to adopt high-speed shearing machine 20000r/min high speed shear 1～3min, make primary emulsion C; Then primary emulsion C is adopted the even method of high pressure breast, respectively 200bar circulation 10 times, 500bar circulation 10 times, 1000bar circulation 20 times makes docetaxel long-circulating target lipid nanometer suspensoid.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN 201210058386 CN102579337B (en) | 2012-03-07 | 2012-03-07 | Long circulation lipid nano-suspension containing docetaxel and preparation method thereof |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN 201210058386 CN102579337B (en) | 2012-03-07 | 2012-03-07 | Long circulation lipid nano-suspension containing docetaxel and preparation method thereof |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN102579337A CN102579337A (en) | 2012-07-18 |
| CN102579337B true CN102579337B (en) | 2013-11-06 |
Family
ID=46468950
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN 201210058386 Expired - Fee Related CN102579337B (en) | 2012-03-07 | 2012-03-07 | Long circulation lipid nano-suspension containing docetaxel and preparation method thereof |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN102579337B (en) |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| EP4299053A1 (en) * | 2022-06-28 | 2024-01-03 | Vysoká skola chemicko-technologická v Praze | Method for preparing an aqueous nanosuspension containing nanocrystals of pharmaceutically active ingredient and a combination of stabilizers, aqueous nanosuspension and use thereof |
Families Citing this family (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103340883A (en) * | 2013-07-25 | 2013-10-09 | 山东大学 | Ceramide-based combined medicine for treating tumor |
| CN103536930B (en) * | 2013-09-24 | 2015-06-24 | 上海纳米技术及应用国家工程研究中心有限公司 | Polylactic acid-polyethylene glycol-tumor penetrating peptide compound, preparation and application thereof |
| CN104208030A (en) * | 2014-08-01 | 2014-12-17 | 中国人民解放军军事医学科学院放射与辐射医学研究所 | Albumin-combined taxol long-circulation nano-particle freeze-dried preparation |
| CN104856955A (en) * | 2015-05-06 | 2015-08-26 | 刘星言 | Docetaxel loaded lipid vesicle drug delivery system and production method thereof |
| CN105412950B (en) * | 2015-12-08 | 2018-06-29 | 上海师范大学 | With MRI/SPECT bimodal image cancer target multifunctional nano probes and preparation and application |
| CN107970447B (en) * | 2017-11-29 | 2020-09-25 | 徐州医科大学 | Lipid-poly-hypoxia radiotherapy sensitizer with dual functions of targeting and radiotherapy sensitization |
Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101317816A (en) * | 2007-06-08 | 2008-12-10 | 中国科学院上海药物研究所 | Docetaxel long-circulation formulation |
| CN101439182A (en) * | 2008-12-18 | 2009-05-27 | 北京大学 | Somatostatin receptor mediated tumor-targeted medicament composition |
| CN101653416A (en) * | 2009-09-22 | 2010-02-24 | 北京大学 | Tumor dual target liposome mediated by integrin and preparation method thereof |
| CN101653414A (en) * | 2008-08-19 | 2010-02-24 | 中国科学院上海药物研究所 | Long-circulating solid lipid docetaxel nanoparticles and preparation method thereof |
| CN101773480A (en) * | 2010-01-19 | 2010-07-14 | 山东大学 | Preparation method of Nnanocrystal preparation containing docetaxelDuoxitasai and method for preparing a freeze-drying agent thereof |
Family Cites Families (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JP5250229B2 (en) * | 2007-09-25 | 2013-07-31 | 独立行政法人科学技術振興機構 | Carbon nanohorn |
| CN101780042A (en) * | 2010-03-26 | 2010-07-21 | 天津大学 | Taxol nano targeting slow-release long-circulating liposome and preparation method thereof |
-
2012
- 2012-03-07 CN CN 201210058386 patent/CN102579337B/en not_active Expired - Fee Related
Patent Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101317816A (en) * | 2007-06-08 | 2008-12-10 | 中国科学院上海药物研究所 | Docetaxel long-circulation formulation |
| CN101653414A (en) * | 2008-08-19 | 2010-02-24 | 中国科学院上海药物研究所 | Long-circulating solid lipid docetaxel nanoparticles and preparation method thereof |
| CN101439182A (en) * | 2008-12-18 | 2009-05-27 | 北京大学 | Somatostatin receptor mediated tumor-targeted medicament composition |
| CN101653416A (en) * | 2009-09-22 | 2010-02-24 | 北京大学 | Tumor dual target liposome mediated by integrin and preparation method thereof |
| CN101773480A (en) * | 2010-01-19 | 2010-07-14 | 山东大学 | Preparation method of Nnanocrystal preparation containing docetaxelDuoxitasai and method for preparing a freeze-drying agent thereof |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| EP4299053A1 (en) * | 2022-06-28 | 2024-01-03 | Vysoká skola chemicko-technologická v Praze | Method for preparing an aqueous nanosuspension containing nanocrystals of pharmaceutically active ingredient and a combination of stabilizers, aqueous nanosuspension and use thereof |
Also Published As
| Publication number | Publication date |
|---|---|
| CN102579337A (en) | 2012-07-18 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN102579337B (en) | Long circulation lipid nano-suspension containing docetaxel and preparation method thereof | |
| CN101474155B (en) | Lung-targeted medicine carrying precursor liposome for injection and method of use thereof | |
| DK2508170T3 (en) | LIPOSOM OF IRINOTECAN OR ITS HYDROCHLORIDE AND ITS PROCEDURE | |
| CN1840193B (en) | Nanometer capsule of anthracene nucleus anticancer antibiotic with polyethylene glycol-phospholipid | |
| US9814734B2 (en) | Bufalin liposome, preparation method therefor and application thereof | |
| US20110052708A1 (en) | Methods and formulations for the delivery of pharmacologically active agents | |
| US20060073175A1 (en) | Methods and formulations for delivery of pharmacologically active agents | |
| US20090196918A1 (en) | Liposomal formulations of hydrophobic lactone drugs in the presence of metal ions | |
| Yang et al. | Cisplatin and paclitaxel co-delivered by folate-decorated lipid carriers for the treatment of head and neck cancer | |
| Fang et al. | SN38-loaded< 100 nm targeted liposomes for improving poor solubility and minimizing burst release and toxicity: in vitro and in vivo study | |
| Xin et al. | PLGA nanoparticles introduction into mitoxantrone-loaded ultrasound-responsive liposomes: In vitro and in vivo investigations | |
| CN102784109B (en) | Taxane medicines albumin nanoparticle preparation for injection and preparation method thereof | |
| CN109771663B (en) | Preparation and application of acid-responsive anticancer nano-drug | |
| CN108567742B (en) | SN38 lipid composition, preparation method and application thereof | |
| CN106139151A (en) | Ascorbic acid palmityl ester and the synergistic pharmaceutical composition of antitumor drug | |
| Zhang et al. | Pharmacokinetics, distribution and anti-tumor efficacy of liposomal mitoxantrone modified with a luteinizing hormone-releasing hormone receptor-specific peptide | |
| CN113384530B (en) | Polysaccharide core Nanocells and preparation method and application thereof | |
| CN100594902C (en) | Nano micelle preparation of Catharanthus roseus alkaloids antineoplastic drugs with coating of phospholipid derived from polyethylene glycol | |
| CN104274401A (en) | High drug-loading content nano suspension for camptothecin medicine based on HCPT-PEG (hydroxycamptothecin-polyethylene glycol) and preparation method of high drug-loading content nano suspension | |
| CN104548125A (en) | Preparation and application of PEG-PTX-NCs (pegylation-paclitaxel-nanocrystals) | |
| Hu et al. | Docetaxel-loaded cholesterol-PEG co-modified poly (n-butyl) cyanoacrylate nanoparticles for antitumor drug pulmonary delivery: preparation, characterization, and in vivo evaluation | |
| CN109381429B (en) | Paclitaxel targeted sustained-release liposome modified by leukocyte membrane and preparation method thereof | |
| CN107158395B (en) | Cabazitaxel phospholipid composition and preparation method and application thereof | |
| CN101322681B (en) | Method for preparing nano micelle formulation of anthracene nucleus antineoplastic antibiotic | |
| CN102085189B (en) | Docetaxel liposome sterile lyophilized preparation and preparation method thereof |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| C14 | Grant of patent or utility model | ||
| GR01 | Patent grant | ||
| CF01 | Termination of patent right due to non-payment of annual fee | ||
| CF01 | Termination of patent right due to non-payment of annual fee |
Granted publication date: 20131106 |