2-thio-n-hydroxycytosine ribonucleoside phosphate and its use in antiviral drugs
A technology of hydroxycytosine ribonucleoside phosphate and thiolation, applied in the field of medicinal chemistry, can solve problems such as long survival period, virus rebound, drug resistance, etc., and achieve high product yield, good inhibitory effect, and simple steps Effect
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Embodiment 1
[0039] The synthetic method of N-hydroxyl 2-thio-N-hydroxycytosine ribonucleoside phosphate, its synthetic route is as follows:
[0040]
[0041] Compound A (10 g), hexamethyldisilazane (300 mL), trimethylchlorosilane (7.71 g) and ammonium sulfate (412 mg) were mixed, N 2Under protection, stir at 126 °C for 18 hours to make the solution clear, concentrate the reaction solution, then drain it with an oil pump, add 1,2-dichloroethane (200 mL), compound B (27.6 g) to the reaction solution, and finally add Tin tetrachloride (27.0 g), stirred at room temperature for 1 h. Add saturated sodium bicarbonate solution dropwise to quench, extract with dichloromethane, wash the organic phase with saturated brine, dry over anhydrous sodium sulfate, filter to remove anhydrous sodium sulfate, concentrate the filtrate, and dry it with an oil pump to obtain compound C (33.7 g , white solid), compound C was directly used in the next step without further purification.
[0042] Compound C (4 ...
Embodiment 2
[0051] Example 2 In vitro anti-HBV activity research
[0052] The prepared compound and the positive control drugs telbivudine and lamivudine were used for in vitro anti-HBV efficacy evaluation test.
[0053] Telbivudine (Shanghai Titan Technology Co., Ltd.), Lamivudine (Shanghai Titan Technology Co., Ltd.), HepG2.2.15 cells (provided by the Antiviral Drug Research Laboratory, School of Pharmacy, Fudan University), fetal bovine serum (FBS) (Thermo Fisher Biochemicals Co., Ltd.), DMEM medium (Thermo Fisher Biochemicals Co., Ltd.), carbon dioxide incubator (Thermo Fisher Biochemicals Co., Ltd.), fluorescent quantitative PCR (Thermo Fisher Murphy Fisher Biochemical Co., Ltd.).
[0054] Detection of drug antiviral activity: take a bottle of well-grown HepG2.2.15 cells, digest with trypsin, make a single cell suspension, count with a cell counting plate, and adjust the cells with DMEM medium containing 10% FBS serum. Density to 2 x 10 5 Individual / mL, inoculated in culture plate...
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