Discovery of notoprenylate F from Notopterygium incisum (Qianghuo) as a PPARγ agonist with potent anti-inflammatory activity through HTS²

Abstract

Notopterygium incisum, a traditional medicinal herb widely used for treating rheumatic pain and inflammatory disorders, is an important source of structurally diverse and pharmacologically active coumarins. Using an HTS²-guided screening strategy, we identified 13 previously undescribed coumarins, notoprenylates A–K (compounds 1–11), including two pairs of enantiomers. The structures of these compounds were elucidated through integrated spectroscopic analyses, including HRESIMS, NMR, and chemical calculations, complemented by Mo₂(AcO)₄-induced circular dichroism experiments. Among these new compounds, notoprenylate F (compound 6) showed mostly inhibition of pro-inflammatory factor NO with a IC₅₀ of 7.01 μmol/L. Transcriptome data from THP-1 cells treated with new compounds collected using the HTS² platform revealed that compound 6 significantly regulates the PPAR signaling pathway, thereby exerting anti-inflammatory effects. Molecular docking and SPR assays suggested that ompound 6 binds to PPARγ, with a K_d value of 5.3 μmol/L. Furthermore, our results showed that compound 6 acts as a PPARγ agonist, and exerts anti-inflammatory effects by inhibiting PPARγ-mediated MAPK, NF-κB, and NLRP3 inhibition. Molecular docking simulations further confirmed that the hydroxyl group and tricyclic epoxide on the side chain form stable interactions with GLN283 and TYR473. This indicates that the isoprenyl side chain of compound 6 is critical for fitting into the pocket of the PPARγ ligand-binding domain (LBD). Overall, this study not only enriches the chemical diversity of coumarins from N. incisum but also identifies notoprenylate F as a promising PPARγ-targeted anti-inflammatory candidate, providing new insights into the pharmacological potential of traditional medicinal plants.

Keywords

HTS² / Coumarin / PPARγ agonist / Anti-inflammatory / Notopterygium incisum

References

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