One-pot synthesis and application of selenite esterified chitosan copper for anti-liver cancer drug
A technology of chitosan copper and selenite, which is applied in the field of biomedical engineering, can solve the problems of low selenization amount, insufficient activity, and failure to achieve a high ratio, so as to reduce dissolution, prevent copper ions and amino groups from degrading, Eliminate the effect of other side effects
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Embodiment 1
[0041] Embodiment 1 One-pot synthesis method of anti-liver cancer drug selenite esterified chitosan copper
[0042] 9.0g chitosan (molecular formula (C 6 h 11o 4 N·H 2 (O)n, equivalent to chitosan monomer 50mmol) is dissolved in 700mL mass fraction 1% formic acid solution, then add the aqueous solution of 12.5g copper sulfate pentahydrate (that is: 50mmol copper sulfate pentahydrate), stir at 60-70 ℃ After reacting for 1 h, adjust the pH value of the reaction solution to 7 with a 10% aqueous sodium hydroxide solution by mass fraction, stir the reaction for 1 h and wait for the pH value to decrease, then adjust the pH value of the reaction solution to 7 with a 10% aqueous sodium hydroxide solution again, Continue the reaction; repeat this for 2-3 times until the pH value no longer drops, and the complexation reaction is complete (the identification method is: take 2mL solution and add 2mL ethanol to precipitate the product, take the supernatant and add dropwise sodium hydrox...
Embodiment 2
[0057] The IR spectrum contrast of the selenite esterified chitosan copper prepared in embodiment 2 chitosan copper and embodiment 1
[0058] Compare chitosan copper [preparation by the method for preparing chitosan copper by literature (Wang Aiqin, etc., the synthesis and characterization of chitosan and Cu (Ⅱ) complexes [J], 2000 (03): 297-300)] and implementation The IR figure of the prepared selenite esterified chitosan copper pure product of example 1, the result is as follows figure 1 and figure 2 as shown, figure 2 It is easy to see in the IR diagram of selenized chitosan copper: compared with figure 1 In the IR spectrum of chitosan copper, the vibration peak of the Se=O double bond of very strong selenite is observed at 713cm-1, the free hydroxyl peak (3390cm-1) disappears, and the free hydroxyl is completely selenous After esterification, there is no influence of free hydroxyl groups, and the complexed hydroxyl groups and hydroxide groups in the compound can clea...
Embodiment 3
[0060] Anti-liver cancer activity test of embodiment 3 selenite esterified chitosan copper
[0061] Human hepatoma cells HepG2 were cultured in DMEM medium containing 10% fetal bovine serum and 100 U / mL of penicillin and streptomycin in a 37°C incubator with a CO2 concentration of 5% [15]. Change the medium every 1-2 days or when the medium turns light yellow. When the cells grew to 80% of the bottom area of the flask, the cells were subcultured. Operate in an ultra-clean bench, first discard the old medium, wash gently with normal saline twice, add an appropriate amount of trypsin-EDTA to digest, observe the cells under the microscope to become round, quickly add new medium, and stop Digestion. Blow down the cells with a pipette gun, transfer to a centrifuge tube, centrifuge at 1 000r / min for 5min, discard the supernatant, resuspend in the new medium, count the cells with a hemocytometer, and pass passage at an appropriate ratio .
[0062] 96-well plate culture: Add Hep...
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