A polymer targeting drug carrier for bladder tumor perfusion therapy and preparation method thereof
A bladder tumor and targeting technology, which is applied in the direction of non-active components of polymer compounds, antineoplastic drugs, drug combinations, etc., can solve the problems of troublesome treatment, limited application, low response rate, etc., and achieves a simple and easy preparation method, good Targeting effect, high drug loading effect
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Embodiment 1
[0042] Embodiment 1: the synthesis of amphiphilic block polymer
[0043] 1) Synthesis of PEO-b-PCL
[0044] Synthetic route such as figure 1 shown. The ethylene oxide monomer (EO) and tetrahydrofuran (THF) solvents were dehydrated and deoxygenated, the reaction device was evacuated three times, protected by argon, and placed in an ice-water bath. Use a double-pointed needle to transfer the cooled and dried EO monomer to the reaction tube, and design the molecular weight according to the experiment. Under the protection of argon, THF and bis(trimethylsilyl)nitrogen potassium [(CH 3 ) 3 Si] 2 The NK initiators are sequentially added to the reaction tube. After reacting for 2-5 days, a THF solution of a certain volume of caprolactone monomer (CL) was added under the protection of argon. After continuing the reaction for 0.5-3 hours, add acetic acid to terminate the reaction, and precipitate the polymer product in excess cold ether to obtain an amphiphilic block copolymer NH...
Embodiment 2
[0055] Example 2: Synthesis of polymers modified by fluorescent molecules and targeting molecules
[0056] 1) Synthesis of FITC-PEO-b-PCL
[0057] Synthetic route such as figure 2 shown. Weigh 0.0780g of fluorescein isothiocyanate (FITC) and dissolve it in 5ml of dimethyl sulfoxide (DMSO), add it to 1.0g of NH 2 -PEO 5k -b-PCL 5k In the reaction tube, react in an oil bath at 20-100°C for 72 hours, dialyze twice in distilled water, centrifuge to remove the upper clear night, and freeze-dry the lower precipitate to obtain the product FITC-PEO 5k -b-PCL 5k . The structure and composition of the product can be determined by GPC and 1HNMR to co-confirm. In the same way, RBITC-PEO-b-PCL can be obtained by replacing FITC with tetramethylrhodamine (RBITC).
[0058] 2) Synthesis of FA-PEO-b-PCL
[0059] Synthetic route such as image 3 shown. 1-Ethyl-(3-dimethylaminopropyl) carbodiimide hydrochloride (EDC, 0.0396g), N-hydroxysuccinimide (NHS, 0.0160g) and folic acid (FA, 0...
Embodiment 4
[0078] Example 4: Targeted adsorption test of micelles on T24 cells
[0079] 1) Cell culture
[0080] Human bladder tumor cell T24 cells were selected for in vitro experiments, and the cells were cultured in F-12 medium (adding 10% fetal bovine serum and 1% penicillin and streptomycin) at 37°C and 5% CO 2 Incubated in the incubator. During the experiment, cells in good growth state were taken at 1×10 5 The density of cells / well was seeded in a 24-well plate. When the cells spread to 70%, discard the old culture medium, rinse with phosphate buffered saline (PBS) three times, add fresh culture medium, and add a certain amount of different concentrations (0.15mg / ml, 0.3mg / ml, respectively) at the same time. ml, 0.45mg / ml) of unmodified CH3O-PEO5k-b-PCL5k fluorescent micelles solution or FA-PEO with dual functions of targeting and fluorescence 5k -b-PCL 5k The micellar solution continued to grow.
[0081] 2) Laser confocal microscope observation
[0082] After the c...
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