Atazanavir sulfate (BMS-232632 sulfate) is an azapeptide and HIV-PROTEASE INHIBITOR that is used in the treatment of HIV INFECTIONS and AIDS in combination with other ANTI-HIV AGENTS.

HIV protease:2.66 nM(Ki)

30 mM Atazanavir作用于LS180V细胞,免疫反应的P-gp表达提高2.5倍,细胞内Rh123降低。Atazanavir抑制U251,T98 g和 LN229胶质母细胞瘤细胞系的细胞生长,显著地提高GRP78和CHOP蛋白水平。Atazanavir也抑制人类20S proteasome 蛋白酶体。Atazanavir作用于U251胶质瘤细胞,各种不同大小的多聚泛素化蛋白质明显增加。Atazanavir 作用于病毒感染的H9细胞,抑制病毒gag前体p55聚合蛋白的蛋白水解裂解,IC50为~47 nM。Atazanavir作用于RF/MT-2 株,具有有效的抗病毒活性,EC50 为3.89 nM。30 μM Atazanavir作用于HepG2细胞,改变内质网应激的幅度和UPR基因表达。

Protease assays: To determine the inhibition constants (Ki) for each Prt inhibitor, purified HIV-1 RF wild-type Prt (2.5 nM) is incubated at 37 ℃ with 1 μM to 15 μM fluorogenic substrate in reaction buffer (1 M NaCl, 1 mM EDTA, 0.1 M sodium acetate [pH 5.5], 0.1% polyethylene glycol 8000) in the presence or absence of Atazanavir. Cleavage of the substrate is quantified by measuring an increase in fluorescent emission at 490 nM after excitation at 340 nM using a Cytofluor 4000. Reactions are carried out using 1.36 μM, 1.66 μM, 2.1 μM, 3.0 μM, 5.0 μM, or 15 μM substrate in the presence of five concentrations of Atazanavir (1.25 nM to 25 nM). Substrate cleavage is monitored at 5-min intervals for 30 min. Cleavage rates are then determined for each sample at early time points in the reaction, and Ki values are determined from the slopes of the resulting Michaelis-Menten plots.

To determine cytotoxicity, host cells are incubated in the presence of serially diluted Atazanavir for 6 days and cell viability is quantitated using an XTT[2,3-bis(2-methoxy-4-nitro-5-sulfophenyl-2H-tetrazolium-5-carboxanilide] assay to calculate the 50% cytotoxic concentrations (CC50s). To assess the effect of human serum proteins on antiviral activity, the 10% fetal calf serum normally used for assays is replaced with 40% adult human serum or 1 mg of α1-acid glycoprotein/mL.(Only for Reference)