Asunaprevir (BMS-650032) is an effective hepatitis C virus (HCV) NS3 protease inhibitor.

4a (ED43):1.6 nM, 1b J4L6S:0.3 nM, 6a (HK-6A):0.9 nM, HCV 1a H77:0.7 nM, 5a (SA13):1.7 nM

Asunaprevir抑制基因型1a（H77株）和基因型1b（J4L6S株）的NS3蛋白酶活性，其IC50分别为0.7和0.3 nM。针对编码NS3蛋白酶领域的基因型1a、1b和4a的复制子，ASV的EC50值在1.2到4.0 nM之间[2]。在以10倍和30倍EC50值（分别为50或150 nM最终浓度）的Asunaprevir浓度下，维持复制子细胞的选择性压力。对于基因型1b的抗性选择，以10倍或30倍EC50值（分别为30或90 nM最终浓度）的Asunaprevir浓度维持复制子细胞[3]。单次或多次给药，每日两次，剂量从200到600 mg的Asunaprevir通常被患者良好耐受，能在慢性感染基因型1 HCV的受试者中快速且大幅降低HCV RNA水平[4]。

Asunaprevir（3-15 mg/kg，p.o.）在多种动物种类中表现出肝脏靶向性质，肝脏至血浆比例在不同物种间从40至359倍不等。在给药后24小时，所有测试物种的肝脏暴露度普遍达到或超过抑制剂EC50（针对HCV基因型1复制子）的110倍以上[2]。

Cytotoxicity is determined by incubating cells (3,000 to 10,000 cells/well) with serially diluted test compounds or DMSO for 5 days (MT-2 cells) or 4 days (all other cell types). Cell viability is quantitated using an MTS assay for MT-2 or a Cell-Titer Blue reagent assay for HEK-293, HuH-7, HepG2, and MRC5 cells, and 50% cytotoxic concentrations (CC50s) are calculated.

Mice (n=9 per group; overnight fast) receive Asunaprevir (ASV) by oral gavage (5 mg/kg; a vehicle of PEG-400-ethanol, 9:1). Blood samples (-0.2 mL) are obtained by retro-orbital bleeding at 0.25, 0.5, 1, 3, 6, 8, and 24 h after dosing. Within each group, three animals are bled at 0.25, 3, and 24 h, three at 0.5 and 6 h, and three at 1 and 8 h, resulting in a composite pharmacokinetic profile. Livers and brains are also removed from mice at the terminal sampling points. Rats (n=3 per group; overnight fast) receive ASV (amorphous free acid) by oral gavage (3, 5, 10, and 15 mg/kg) in PEG-400-ethanol (9:1). Serial blood samples (-0.3 mL) are obtained from the jugular vein predosing (0 h) and at 0.25, 0.5, 0.75, 1, 2, 4, 6, 8, 24, and 48 h post dosing. To assess tissue exposure, rats are orally administered ASV (5 or 15 mg/kg, same vehicle as above), and blood, liver, and heart samples from two rats/group are obtained at 0.17, 0.5, 1, 2, 4, 6, 8, 24, 48, and 72 h after dosing.