Preparation method of cisplatin nano medicament for treating ovarian cancer
A nano-drug, cisplatin technology, applied in the field of ovarian cancer, can solve the problem of no significant improvement in the progression-free survival of patients, and achieve the effect of inhibiting proliferation and metastasis, and reducing toxic and side effects
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Embodiment 1
[0020] Example 1: Preparation and Stability Detection of Cisplatin Nanomedicine
[0021] 1. Preparation of cisplatin nanomedicine
[0022] Take 50 mg of carboxylated heparin, 6 mg of folic acid, and 6 mg of cRGD and dissolve them in 3 mL of dimethyl sulfoxide; then add 21 mg of 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide, N-hydroxysuccinyl 15 mg of amine was reacted at room temperature for 12 hours, dialyzed in deionized water for 48 hours, and concentrated to obtain a carboxylated heparin-folate-cyclopeptide nanoparticle solution. The concentration of folic acid in carboxylated heparin-folic acid-cyclic peptide solution detected by ultraviolet spectrophotometer is 0.36mg / mL; the concentration of cRGD in carboxylated heparin-folic acid-cyclic peptide solution is detected by CQBCA method is 0.05mg / mL; lyophilized 1mL of carboxylated heparin-folate-cyclopeptide solution weighs 2.5mg, and subtracting the mass of folic acid and cRGD, the mass of carboxylated heparin can be obta...
Embodiment 2
[0025] Example 2: Cytotoxicity experiment of cisplatin nanomedicine on ovarian cancer cells SKOV3 and A2780
[0026] SKOV3 and A2780 cells were cultured to the logarithmic growth phase, seeded into a 96-well plate, and the number of cells per well was about 3000. After routine culture overnight, 5 concentration gradients were set with cisplatin as the quantitative standard, namely 2 μg / mL, 4 μg / mL, 8μg / mL, 16μg / mL, 32μg / mL act on the cells, and set 5 duplicate wells for each concentration. In the control group, only the medium was replaced. The absorbance of each hole at 490nm was detected by a standard instrument, and the relative cell survival rate of each group was calculated according to the formula (%)=(average absorbance value of the experimental group / average absorbance value of the control group)×100%, and the results were as follows: image 3 The results showed that the inhibitory ability of cisplatin nanomedicine on the proliferation of ovarian cancer tumor cells SK...
Embodiment 3
[0027] Embodiment 3: In vivo biological function experiment of cisplatin nanomedicine
[0028] 1. Distribution of cisplatin nanomedicine in nude mouse orthotopic ovarian cancer model
[0029]The SKOV3 (SKOV3-Luc) cell line labeled with firefly luciferase was selected to construct a nude mouse ovarian carcinoma in situ model, and the tumor fluorescence was detected with an in vivo imager; the prepared cisplatin nanomedicine was labeled with cy7, and the cisplatin 2 μg / The dose of g was injected into the tail vein of nude mice, and the cy7 fluorescence of the drug was detected with an in vivo imager at 30 minutes, 3 hours, 6 hours, 12 hours, and 24 hours after injection; the mice were dissected after 24 hours, and the heart was taken , liver, spleen, lung, kidney, and tumor tissue to detect drug cy7 fluorescence and tumor fluorescence. The result is as Figure 4 As shown, the fluorescence of pure cy7 decays rapidly within 24 hours, while the fluorescence value of cisplatin na...
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