Multi-layered target identification strategy of natural products: Daphnetin targets NQO1 and OPLAH in silicosis alleviation

Ling Wang; Bo-Yan Ma; Sheng-Ping Jiang; Shu-Ting Wei; Xue-Mei Qin; Zhen-Yu Li

doi:10.1016/j.jpha.2026.101629

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Ling Wang, Bo-Yan Ma, Sheng-Ping Jiang, Shu-Ting Wei, Xue-Mei Qin, Zhen-Yu Li. Multi-layered target identification strategy of natural products: Daphnetin targets NQO1 and OPLAH in silicosis alleviation[J]. Journal of Pharmaceutical Analysis. doi: 10.1016/j.jpha.2026.101629

Citation:

Ling Wang, Bo-Yan Ma, Sheng-Ping Jiang, Shu-Ting Wei, Xue-Mei Qin, Zhen-Yu Li. Multi-layered target identification strategy of natural products: Daphnetin targets NQO1 and OPLAH in silicosis alleviation[J]. Journal of Pharmaceutical Analysis. doi: 10.1016/j.jpha.2026.101629

Citation:

Ling Wang, Bo-Yan Ma, Sheng-Ping Jiang, Shu-Ting Wei, Xue-Mei Qin, Zhen-Yu Li. Multi-layered target identification strategy of natural products: Daphnetin targets NQO1 and OPLAH in silicosis alleviation[J]. Journal of Pharmaceutical Analysis. doi: 10.1016/j.jpha.2026.101629

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Multi-layered target identification strategy of natural products: Daphnetin targets NQO1 and OPLAH in silicosis alleviation

doi: 10.1016/j.jpha.2026.101629

Ling Wang^a,b,
Bo-Yan Ma^a,b,
Sheng-Ping Jiang^a,b,
Shu-Ting Wei^c,d,
Xue-Mei Qin^a,b,
Zhen-Yu Li^{a,b
,
,}

a. Modern Research Center for Traditional Chinese Medicine, Shanxi University, Taiyuan, 030006, China;
b. The Key Laboratory of Chemical Biology and Molecular Engineering of Ministry of Education, Shanxi University, Taiyuan, 030006, China;
c. First Clinical Medical College, Shanxi Medical University, Taiyuan, 030000, China;
d. National Health Commission Key Laboratory of Pneumoconiosis, Shanxi Key Laboratory of Respiratory Diseases, Department of Pulmonary and Critical Care Medicine, The First Hospital of Shanxi Medical University, Taiyuan, 030000, China

Funds:

This work was supported by the National Natural Science Foundation of China (Grant No.: 81973466), the National Administration of Traditional Chinese Medicine Youth Qihuang Scholars Support Project, China, and the Program of Graduate Innovation Research in Shanxi Province, China (Program No.: 2023KY019). The authors would like to acknowledge BioGDP.com (https://biogdp.com/) for providing the platform to create the schematic diagrams in this article.

Received Date: Jul. 23, 2025
Accepted Date: Apr. 02, 2026
Rev Recd Date: Apr. 02, 2026
Available Online: Apr. 03, 2026

Abstract

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).
- Silicosis,
- Daphnetin,
- Multi-layered target identification,
- NQO1,
- OPLAH

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References(37)

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