Application of combination of dianhydrogalactitol and autophagy inhibitor in anti-melanoma drug preparation
An autophagy inhibitor and anti-melanin technology, applied in the field of medicine, can solve the problems of no research on the role of melanoma, impaired immune function of patients, and decreased quality of life. hysteresis effect
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Embodiment 1
[0024] The application of Weikangol and an autophagy inhibitor in combination in the preparation of an anti-melanoma drug, the anti-melanoma drug comprising Weikangol and an autophagy inhibitor.
[0025] Further, the chemical formula of Weiconol is:
[0026]
[0027] Further, the autophagy inhibitor is selected from 3-MA.
[0028] Further, the anti-melanoma drug is a compound drug made of Weikangol and an autophagy inhibitor.
[0029] Further, the compound drug is prepared in the form of solid, solution, dispersion, micelle, emulsion, liposome or nano-microspheres.
[0030] Further, the autophagy inhibitor in the compound drug and Weikangol are administered in combination or sequentially.
[0031] Further, the autophagy inhibitor inhibits the autophagy of the melanoma cells induced by Weiconol, and the antagonism of the melanoma cells to Weiconol treatment caused by the autophagy.
Embodiment 2
[0033] Preparation of main reagents:
[0034] Preparation of trypsin: Weigh KCl0.4g, KH2PO40.06g, NaCl8.0g, NaHCO30.35g, NaH:PO412H2O0.08g, phenol red 0.02g, EDTANa0.255g, dissolve in about 800ml deionized water in turn, adjust the pH to 7.2-7.4, finally add 2.5g of trypsin, add water to make up to 1L, make the trypsin concentration 0.25%, adjust the pH to about 7.4, carry out filter sterilization, store at 4°C after aliquoting.
[0035] Preparation of Weikangol solution: Accurately weigh 0.2mg of Weikangol in a 0.2ml EP tube, add 4μl of DMSO to dissolve it, and then make 50mg / ml Weikangol mother solution, and store it in the dark at 4°C. Dilute to the required concentration with serum-free medium or complete medium before use.
[0036] Preparation of autophagy inhibitor solution: Dissolve an appropriate amount of 3-MA in PBS to make a 10 mg / mL storage solution, filter and sterilize it with a 0.1 μm filter, and pack it into 100 μL / bottle, one at a time, Subcutaneous or intra...
Embodiment 3
[0040] Cell culture: Use PRMI-1640 dry powder medium to dissolve B16F0 cells in 1000ml of fresh three-distilled water. After complete dissolution, add penicillin and streptomycin to make the final concentration 100U / mL, 100μg / mL, and finally add 3.7gNaHCO3, Adjust the pH to 7.2-7.4 and stir thoroughly, then filter and sterilize, aliquot and store at 4°C for culture, when the cells grow to the logarithmic growth phase, digest with trypsin, digest the cells at 1*105cells / The concentration of mL was transferred to the cell culture plate for culture, and after 12 hours of culture, the cells in each group were given the corresponding concentration of Viconol solution; in the case of combined administration, 2mm3-MA was added 1 hour before administration to inhibit the autophagy of B16F0 cells; continuous culture After 24h, 48h or 72h, proceed to the next step.
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