Department of Microbiology, Faculty of Science, Mahidol University, Bangkok 10400, Thailand.
Laboratory of Medicinal Chemistry, Chulabhorn Research Institute, Bangkok 10210, Thailand.
Viruses. 2023 Sep 27;15(10):2012. doi: 10.3390/v15102012.
Every year, dengue virus (DENV) affects millions of people. Currently, there are no approved drugs for the treatment of DENV infection. Autophagy is a conserved degradation process that was shown to be induced by DENV infection and required for optimal DENV replication. The modulation of autophagy is, therefore, considered an attractive target to treat DENV infection. This study carried out a high-content image screen analysis using Crispr-Cas9 GFP-LC3 knocked-in HeLa cells of a compound library synthesized from or inspired by natural products and their biocongener precursors to discover novel autophagy inhibitors. The screen identified Ka-003 as the most effective compound for decreasing the number of autophagic vacuoles inside cells upon autophagy induction. Ka-003 could inhibit autophagy in a dose-dependent manner at low micromolar concentrations. More importantly, Ka-003 demonstrated the concentration-dependent inhibition of DENV production in Crispr-Cas9 GFP-LC3 knocked-in THP-1 monocytes. The core structure of Ka-003, which is a methyl cyclohexene derivative, resembles those found in mulberry plants, and could be synthetically prepared in a bioinspired fashion. Taken together, data indicate that Ka-003 hampered autophagy and limited DENV replication. The low cytotoxicity of Ka-003 suggests its therapeutic potential, which warrants further studies for the lead optimization of the compound for dengue treatment.
每年，登革热病毒（DENV）都会影响数百万人。目前，尚无批准的药物可用于治疗 DENV 感染。自噬是一种保守的降解过程，已被证明可被 DENV 感染诱导，并对最佳 DENV 复制至关重要。因此，自噬的调节被认为是治疗 DENV 感染的一个有吸引力的目标。本研究使用 Crispr-Cas9 GFP-LC3 敲入 HeLa 细胞进行了高内涵图像筛选分析，该细胞使用从天然产物或其生物同源物前体合成的化合物库进行了筛选，以发现新型自噬抑制剂。筛选确定 Ka-003 是在自噬诱导时降低细胞内自噬小泡数量的最有效化合物。Ka-003 可以在低微摩尔浓度下以剂量依赖性方式抑制自噬。更重要的是，Ka-003 证明了在 Crispr-Cas9 GFP-LC3 敲入 THP-1 单核细胞中，DENV 产生的浓度依赖性抑制。Ka-003 的核心结构是甲基环己烯衍生物，类似于桑植物中的结构，并且可以以仿生的方式进行合成制备。总而言之，数据表明 Ka-003 阻碍了自噬并限制了 DENV 的复制。Ka-003 的低细胞毒性表明其具有治疗潜力，值得进一步研究以优化该化合物用于登革热治疗的先导化合物。
