Validation and Characterization of Five Distinct Novel Inhibitors of Human Cytomegalovirus

The
critical consequences of human cytomegalovirus (HCMV) infection
in the transplant population and in congenitally infected infants,
the limited treatment options for HCMV, and the rise of resistant
mutants toward existing therapies has fueled the search for new anti-HCMV
agents. A pp28-luciferase recombinant HCMV was used as a reporter
system for high-throughput screening of HCMV inhibitors. Approximately
400 000 compounds from existing libraries were screened. Subsequent
validation assays using resynthesized compounds, several virus strains,
and detailed virology assays resulted in the identification of five
structurally unique and selective HCMV inhibitors, active at sub to
low micromolar concentrations. Further characterization revealed that
each compound inhibited a specific stage of HCMV replication. One
compound was also active against herpes simplex virus (HSV1 and HSV2),
and another compound was active against Epstein–Barr virus
(EBV). Drug combination studies revealed that all five compounds were
additive with ganciclovir or letermovir. Future studies will focus
on optimization of these new anti-HCMV compounds along with mechanistic
studies.

人类巨细胞病毒（HCMV）感染在移植人群和先天性感染婴儿中的严重后果，HCMV治疗选择的局限性，以及现有疗法中耐药突变体的出现，这些因素共同推动了新型抗HCMV药物的研发。本研究采用pp28-荧光素酶重组HCMV作为HCMV抑制剂高通量筛选的报告系统。对现有化合物库中的约40万种化合物进行了筛选。随后，利用重新合成的化合物、多种病毒株和详细的病毒学检测进行的验证实验，成功鉴定出五种结构独特且具有选择性的HCMV抑制剂，这些抑制剂在亚微摩尔至低微摩尔浓度范围内具有活性。进一步的表征表明，每种化合物都能抑制HCMV复制的特定阶段。其中一种化合物对单纯疱疹病毒（HSV1和HSV2）也具有活性，另一种化合物对EB病毒（EBV）也具有活性。药物组合研究显示，这五种化合物与更昔洛韦或勒特莫维尔的组合均呈相加作用。未来的研究将聚焦于这些新型抗HCMV化合物的优化及其作用机制的深入研究。