Patick A K, Binford S L, Brothers M A, Jackson R L, Ford C E, Diem M D, Maldonado F, Dragovich P S, Zhou R, Prins T J, Fuhrman S A, Meador J W, Zalman L S, Matthews D A, Worland S T
Agouron Pharmaceuticals, Inc., San Diego, California 92121, USA.
Antimicrob Agents Chemother. 1999 Oct;43(10):2444-50. doi: 10.1128/AAC.43.10.2444.
AG7088 is a potent, irreversible inhibitor of human rhinovirus (HRV) 3C protease (inactivation rate constant (k(obs)/[I]) = 1,470,000 +/- 440,000 M(-1) s(-1) for HRV 14) that was discovered by protein structure-based drug design methodologies. In H1-HeLa and MRC-5 cell protection assays, AG7088 inhibited the replication of all HRV serotypes (48 of 48) tested with a mean 50% effective concentration (EC(50)) of 0.023 microM (range, 0.003 to 0.081 microM) and a mean EC(90) of 0.082 microM (range, 0.018 to 0.261 microM) as well as that of related picornaviruses including coxsackieviruses A21 and B3, enterovirus 70, and echovirus 11. No significant reductions in the antiviral activity of AG7088 were observed when assays were performed in the presence of alpha(1)-acid glycoprotein or mucin, proteins present in nasal secretions. The 50% cytotoxic concentration of AG7088 was >1,000 microM, yielding a therapeutic index of >12,346 to >333,333. In a single-cycle, time-of-addition assay, AG7088 demonstrated antiviral activity when added up to 6 h after infection. In contrast, a compound targeting viral attachment and/or uncoating was effective only when added at the initiation of virus infection. Direct inhibition of 3C proteolytic activity in infected cells treated with AG7088 was demonstrated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis analysis of radiolabeled proteins, which showed a dose-dependent accumulation of viral precursor polyproteins and reduction of processed protein products. The broad spectrum of antiviral activity of AG7088, combined with its efficacy even when added late in the virus life cycle, highlights the advantages of 3C protease as a target and suggests that AG7088 will be a promising clinical candidate.
AG7088是一种强效、不可逆的人鼻病毒（HRV）3C蛋白酶抑制剂（对于HRV 14，失活速率常数（k(obs)/[I]）= 1,470,000 ± 440,000 M⁻¹ s⁻¹），它是通过基于蛋白质结构的药物设计方法发现的。在H1-HeLa和MRC-5细胞保护试验中，AG7088抑制了所有测试的HRV血清型（48种中的48种）的复制，平均50%有效浓度（EC(50)）为0.023微摩尔（范围为0.003至0.081微摩尔），平均EC(90)为0.082微摩尔（范围为0.018至0.261微摩尔），以及包括柯萨奇病毒A21和B3、肠道病毒70和艾柯病毒11在内的相关微小核糖核酸病毒的复制。当在含有α(1)-酸性糖蛋白或粘蛋白（鼻分泌物中的蛋白质）的情况下进行试验时，未观察到AG7088的抗病毒活性有显著降低。AG7088的50%细胞毒性浓度>1,000微摩尔，治疗指数>12,346至>333,333。在单周期、添加时间试验中，AG7088在感染后长达6小时添加时仍表现出抗病毒活性。相比之下，一种靶向病毒附着和/或脱壳的化合物仅在病毒感染开始时添加才有效。通过对放射性标记蛋白质的十二烷基硫酸钠-聚丙烯酰胺凝胶电泳分析，证明了AG7088对感染细胞中3C蛋白水解活性的直接抑制，该分析显示病毒前体多聚蛋白呈剂量依赖性积累，加工后的蛋白质产物减少。AG7088广泛的抗病毒活性，加上即使在病毒生命周期后期添加也具有疗效，突出了3C蛋白酶作为靶点的优势，并表明AG7088将是一个有前景的临床候选药物。
