Aleuritopteris argentea (S. G. Gmél.) Fée is a medicinal fern consisting of an ent -labdane diterpene, i.e. alepterolic aicd, as the major metabolite. We recently isolated grams of alepterolic acid from A. argentea enabling subsequent structural modification. By incorporation of amino moiety to alepterolic acid, fifteen amide derivatives were synthesized, characterized, and further biological evaluated regarding their activity against four cancer cells and normal human liver cells. The potency of synthesized amides dramatically improved as compared to alepterolic aicd itself. The best hit (compound 11 ) inhibits HeLa cells with an IC 50 of 7.39 ± 0.80 μM, and is nearly nontoxic to normal cells. Compound 11 exhibits an inhibitory effect on the colony forming ability of the four cancer cells, especially of HeLa cells. Moreover, it induces apoptosis of HeLa cells by decreasing mitochondrial membrane potential and altering expression of apoptosis-associated proteins. Release of cytochrome c , activation of caspases-3, caspases-9 and alteration of Bax/Bcl-2 balance was detected in the biological assays. These results imply that compound 11 can inhibit the proliferation of cervical cancer cell line HeLa and induce apoptosis through the mitochondrial pathway. These findings encourage further rational structural modification of 15- carboxyl group of alepterolic acid.
Alepterolic acid is a natural compound occurring in Aleuritopteris argentea (Gmél.) Fée. In this study, 12 derivatives of alepterolic acid were obtained by chemical modification and evaluated regarding their effects on nitric oxide (NO) production in RAW264.7 macrophages stimulated by lipopolysaccharide (LPS). Notably, compound 13 exhibited low toxicity while significantly inhibiting LPS-induced NO production. To elucidate its immunomodulatory mechanism, we quantified transcription levels of key cellular inflammatory factors (TNF-α and IL-6) using RT-qPCR and measured protein levels of key cellular inflammatory factor (iNOS) by Western blotting. Additionally, mitochondrial membrane potential and concentrations of reactive oxygen species (ROS) in RAW264.7 cells were analyzed by flow cytometry, and the phosphorylation status of key proteins in the MAPK inflammatory pathway was assessed by Western blotting. The results demonstrate that compound 13 exerts its anti-inflammatory effects by inhibiting MAPK pathway phosphorylation, downregulating LPS-induced transcription of proinflammatory cytokines (TNF- α , IL-6), reducing ROS generation, and delaying the decline of mitochondrial membrane potential. This study provides a basis for the application of alepterolic derivatives in inflammatory diseases.
危险品化学品经营许可证（带存储）