Polymer gene medicine carrier, preparation method thereof and use thereof for preparing antitumor medicine
A gene drug and polymer technology, applied in anti-tumor drugs, gene therapy, drug combination, etc., can solve safety problems and other problems, achieve the effect of good cancer cells and tumors, inhibition of cancer cells and tumors, and low toxicity
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Embodiment 1
[0032] Synthesis of macromolecule gene drug carrier targeting tumors with polyethylenimine as the backbone of hydroxypropyl cyclodextrin as branched chain folic acid:
[0033]
[0034] Dissolve 0.51g of hydroxypropyl cyclodextrin in 6ml of DMSO, add 0.1ml of triethylamine, and dissolve 0.42g of carbonyldiimidazole in 6ml, mix and stir for 1-3 hours under nitrogen protection in the dark. The product is hydroxypropyl cyclodextrin-carbonyldiimidazole. The product was added with 4° ether to produce a precipitate, which was filtered and dissolved in DMSO to form a hydroxypropyl cyclodextrin-carbonyldiimidazole DMSO solution. Dissolve 0.89g of polyethyleneimine with a molecular weight of 600 (CAS No. 9002-98-6) in DMSO, add the above hydroxypropyl cyclodextrin-carbonyldiimidazole DMSO solution dropwise, and place the mixture in a nitrogen-protected light-proof container Stirring was continued for 3-6 hours. The reacted product was placed in a dialysis bag with a molecular weigh...
Embodiment 2
[0037] Cytotoxicity tests and results of the compounds prepared in Example 1.
[0038] Cell lines and culture conditions: HepG2 (human liver cancer cells), Hela (human cervical cancer cells), HLF (human lung fibroblast-like cells), A549 (human lung cancer cells) were provided by the Experimental and Animal Center of Sun Yat-sen University. The cells were cultured in DMEM medium containing 10% fetal bovine serum, which contained 100 units of penicillin and 100 micrograms of streptomycin per milliliter, and the cells were seeded in a 10 cm diameter petri dish at 37 degrees, 5% CO 2 The cells were cultured in the environment, and passaged by trypsinization when the cells were confluent.
[0039] Cytotoxicity test: Cytotoxicity was measured by MTT method. The cells were digested with 0.25% trypsin into a single cell suspension, and the number of viable cells was calculated using a hemocytometer version, and the concentration of viable cells was adjusted to 5 × 10 4 Inoculate eac...
Embodiment 3
[0042] The polymer gene carrier prepared in Example 1 carried gene drug siRNA to inhibit tumor growth test and results.
[0043] Tumors were induced in nude mice first, and in order to obtain tumors, 50 μl of density was 10 6 HeLa cells were injected into 6-7 week old female nude mice with PBS solution. The tumor size was measured every day after the injection of cancer cells. The tumor size was measured with a vernier caliper. The length is (a) and the width is (b). The formula for calculating the tumor volume is V=0.5a×b 2 . When the tumor grows to 80mm 3 The administration was started at the beginning of time, and divided into four administration groups: (1) PBS as control; (2) no therapeutic effect siRNA / folate targeting carrier group; (3) treatment siRNA / no folic acid targeting carrier group; (4) treatment siRNA / folate targeting vector set. Administration was performed once every three days, and the administration method was tail vein injection, and each nude mouse w...
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