Design, synthesis, and antiviral activity evaluation of the bis-ester prodrug of N4-hydroxycytidine phosphate

IF 2.3 4区医学 Q3 CHEMISTRY, MEDICINAL

Bioorganic & Medicinal Chemistry Letters Pub Date : 2026-04-01 Epub Date: 2025-12-25 DOI:10.1016/j.bmcl.2025.130525

Hao Sun , Xu Yan , Chunhuan Mo , Jia Chen , Yan Liu , Yaohui Liu , Xia Han , Jianqin Gao , Lehua Yin , Jinghua Xu , Yuan Chen , Xinhua He

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Abstract

Monophosphorylation is a key rate-limiting step for the efficacy of broad-spectrum nucleoside antiviral drugs, and designing phosphate prodrugs is an effective method to enhance antiviral efficacy. N4-hydroxycytidine (NHC) has a broad-spectrum antiviral effect. Based on the mechanism of action and metabolic inactivation characteristics of NHC, we designed a series of dual ester prodrugs targeting the N4-hydroxyl group and the 5′-phosphate of the ribose ring. Twenty-nine compounds were designed and synthesized. The cytotoxicity assay results showed low cytotoxicity (no significant toxicity at 160 μM). Using the Vesicular Stomatitis Virus expressing Green Fluorescent Protein (VSV-GFP) virus screen, it was found that all of them have antiviral activity. Compounds 2328 and 2322 showed better antiviral activity than molnupiravir. The EC₅₀s of 2328 against VSV, HMPV-A2, and RSV-A2 viruses were 2.19 μM, 35.6 μM, and 53.5 μM, respectively, and the EC₅₀s of 2322 against VSV, HMPV-A2, and RSV-A2 were 6.23 μM, 52.8 μM, and 28.2 μM, respectively. Structure-activity relationships indicate that the antiviral activity of compounds at the cellular level is closely linked to the structure of the prodrug's phosphate ester, whereas the ester group on the N4-hydroxyl is more tolerant of diverse functional groups.

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磷酸n4 -羟基胞苷双酯前药的设计、合成及抗病毒活性评价。

单磷酸化是广谱核苷类抗病毒药物药效的关键限速步骤，设计磷酸原药是提高抗病毒药效的有效方法。n4 -羟基胞苷（NHC）具有广谱抗病毒作用。基于NHC的作用机制和代谢失活特点，我们设计了一系列针对NHC的n4 -羟基和核糖环的5′-磷酸的双酯前药。设计并合成了29个化合物。细胞毒性实验结果显示细胞毒性较低（160 μM无明显毒性）。通过表达绿色荧光蛋白的水疱性口炎病毒（VSV-GFP）病毒筛选，发现它们均具有抗病毒活性。化合物2328和2322的抗病毒活性优于molnupiravir。2328对VSV、HMPV-A2和RSV-A2病毒的ec50分别为2.19 μM、35.6 μM和53.5 μM， 2322对VSV、HMPV-A2和RSV-A2病毒的ec50分别为6.23 μM、52.8 μM和28.2 μM。构效关系表明，化合物在细胞水平上的抗病毒活性与前药的磷酸酯结构密切相关，而n4 -羟基上的酯基对多种官能团的耐受性更强。

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来源期刊

Bioorganic & Medicinal Chemistry Letters 医学-医药化学

CiteScore

5.70

自引率

3.70%

发文量

463

审稿时长

27 days

期刊介绍： Bioorganic & Medicinal Chemistry Letters presents preliminary experimental or theoretical research results of outstanding significance and timeliness on all aspects of science at the interface of chemistry and biology and on major advances in drug design and development. The journal publishes articles in the form of communications reporting experimental or theoretical results of special interest, and strives to provide maximum dissemination to a large, international audience.