Broad-spectrum inhibition of diverse enveloped RNA viruses by the host-directed ZJ-101 analogue series

The global burden of pathogenic enveloped RNA viruses, including flaviviruses, alphaviruses, filoviruses, and orthomyxoviruses, remains a severe public health challenge. Direct-acting antivirals are frequently limited by narrow therapeutic windows and rapid emergence of drug-resistant variants, highlighting an urgent need for host-directed therapeutic strategies. Here, we characterize the ZJ-101 analogue series, a class of small molecules derived from the marine natural product Superstolide A. We demonstrate that ZJ-101 analogues exhibit potent antiviral activity against dengue virus serotype 2 (DENV-2) with EC50 values in the single-digit nanomolar range across orthogonal assay formats and multiple cell types, with minimal host toxicity. This activity extends across diverse RNA viruses, including Zika virus, chikungunya virus, rabies virus, Ebola virus, and multiple influenza A subtypes, consistent with a broad-spectrum host-directed mechanism. Antiviral efficacy was further validated in a physiologically relevant 3D human cerebral organoid model, where ZJ-114 reduced DENV-2 and Zika virus infection to near-undetectable levels at 5 nM. Global quantitative proteomics identified a coordinated reduction in host factors involved in viral attachment, endosomal uptake, glycosylation, and endomembrane trafficking, providing a mechanistic basis for the antiviral breadth. Together, these findings establish the ZJ-101 series as a highly potent, broad-spectrum antiviral scaffold with translational evidence in human tissue models, representing a promising host-directed strategy against endemic and emerging RNA viral threats.