Current drugs for treating human cytomegalovirus (HCMV) infections are limited by resistance and treatment-associated toxicities. In developing mechanistically novel HCMV antivirals, we discovered an N-benzyl hydroxypyridone carboxamide antiviral hit (8a) inhibiting HCMV in submicromolar range. We describe herein the structure–activity relationship (SAR) for 8a, and the characterization of potent analogs for cytotoxicity/cytostatic property, the preliminary mechanism of action, and the absorption, distribution, metabolism and excretion (ADME) properties. The SAR revealed a few pharmacophore features conferring optimal antiviral profile, including the 5-OH, the N-1 benzyl, at least one –CH₂− in the linker, and a di-halogen substituted phenyl ring in the amide moiety. In the end, we identified numerous analogs with sub-micromolar antiviral potency and good selectivity index. The preliminary mechanism of action characterization used a pUL89-C biochemical endonuclease assay, a virus entry assay, a time-of-addition assay, and a compound withdrawal assay. ADME profiling measuring aqueous solubility, plasma and liver microsomal stability, and parallel artificial membrane permeability assay (PAMPA) permeability demonstrated largely favorable drug-like properties. Together, these studies validate the N-benzyl hydroxypyridone carboxamide as a viable chemotype for potent and mechanistically distinct antivirals against HCMV.

目前用于治疗人类巨细胞病毒 (HCMV) 感染的药物受到耐药性和治疗相关毒性的限制。在开发机械新颖的 HCMV 抗病毒药物时，我们发现了N-苄基羟基吡啶酮甲酰胺抗病毒药物 ( 8a ) 在亚微摩尔范围内抑制 HCMV。我们在此描述了8a的构效关系 (SAR) ，以及有效类似物的细胞毒性/细胞抑制特性、初步作用机制以及吸收、分布、代谢和排泄 (ADME) 特性的表征。SAR 揭示了一些具有最佳抗病毒特性的药效团特征，包括 5-OH、N-1 苄基、至少一个 –CH ₂- 在接头中，以及在酰胺部分中的二卤素取代的苯环。最后，我们鉴定了许多具有亚微摩尔抗病毒效力和良好选择性指数的类似物。初步的作用机制表征使用了 pUL89-C 生化核酸内切酶测定、病毒进入测定、添加时间测定和化合物撤除测定。ADME 分析测量水溶性、血浆和肝微粒体稳定性以及平行人工膜通透性测定 (PAMPA) 通透性证明了很大程度上有利的药物样特性。总之，这些研究验证了N-苄基羟基吡啶酮甲酰胺是一种可行的化学类型，可用于有效且机制上不同的抗 HCMV 抗病毒药物。