ArticleJournal of pharmaceutical analysis2026
Multi-layered target identification strategy of natural products: Daphnetin targets NQO1 and OPLAH in silicosis alleviation.
Article in Journal of pharmaceutical analysis, 2026. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Not yet cited in PubMed.
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Abstract
Pneumoconiosis is a common occupational lung disease with unclear pathogenesis and no effective targeted therapies. Emerging evidence suggests that daphnetin is a promising therapeutic candidate for silicosis, yet its efficacy targets remain poorly characterized. In this study, we propose and implement a multi-layered target identification strategy that integrates transcriptomics, clinical metabolomics, public database mining, an artificial intelligence (AI)-based target prediction model, and tissue-based thermal proteome profiling (Tissue-TPP) technology. This strategy enables stepwise elucidation, from pneumoconiosis-related targets to daphnetin efficacy targets and ultimately to direct targets. Using this approach, we validated nicotinamide adenine dinucleotide phosphate hydrogen (NAD(P)H) quinone oxidoreductase 1 (NQO1) and 5-oxoprolinase (OPLAH) as direct targets of daphnetin, and identified Kelch-like ECH-associated protein 1 (KEAP1), nuclear factor erythroid 2-related factor 2 (NRF2), and arginase 1 (ARG1) as indirect targets. Notably, OPLAH is reported here as a direct target of daphnetin for the first time. Furthermore, the binding of daphnetin to NQO1 and OPLAH helps explain its effects on cellular thiol levels. Using a thiol-responsive probe, we observed that daphnetin significantly increased intracellular thiol levels and reduced silica-induced reactive oxygen species (ROS) accumulation in both A549 and THP-1 cells, thereby contributing to its antioxidant effects. Additionally, overexpression (OE) of OPLAH significantly increased reduced glutathione (GSH) levels and alleviated silica-induced epithelial-mesenchymal transition (EMT). Conversely, OPLAH knockdown partially inhibited the efficacy of daphnetin. Together, these findings clarify the anti-fibrotic mechanism of daphnetin, support its therapeutic potential for silicosis, and provide a practical framework for the target discovery of natural products (NPs).
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