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A natural compound derivative P-13 inhibits STAT3 signaling by covalently inhibiting Janus kinase 2

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We investigated the function and molecular mechanisms of 2-desoxy-4β-propylcarbamate-pulchellin (P-13), a sesquiterpene lactone derivative of 2-desoxy-4-epi-pulchellin from the traditional Chinese medicinal herb Carpesium abrotanoides L, in regulating STAT3 signaling and cancer cell growth. We found that P-13 inhibited the IL-6-induced, as well as the constitutive, STAT3 activation in a dose and time-dependent manner. In vitro kinase activity analyses demonstrated that P-13 directly inhibited JAK2 kinase activity. The inhibitory effects of P-13 on JAK2/STAT3 signaling could be blocked by reducing agents dithiothreitol (DTT) or glutathione (GSH), indicating an involvement of the thiol-reactive α-β unsaturated carbonyl group in P-13. Further analyses with mass spectrograph, as well as molecular docking, revealed that P-13 covalently bound with the C452 in the SH2 domain of JAK2. Furthermore, P-13 inhibited growth and induced death of many cancer cell lines, particularly those expressing constitutively activated STAT3. It also inhibited in vivo growth of human cancer cell xenografts. Taken together, these findings revealed P-13 as a novel covalent inhibitor of JAK2, uncovered a new mechanism to inhibit JAK2, and provided a promising anti-cancer drug candidate.

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Acknowledgments

We thank Prof. Xinyuan Fu (National University of Singapore) for providing the HepG2/STAT3 cell line. We also thank Mingwei Sun (Shanghai Institute of Materia Medica) for the help of LC-MS analysis and Yuqi Yu (Shanghai Institute of Materia Medica) for the help of the molecular docking.

Funding

This work was supported by the National Natural Science Foundation of China (grant 81673465 to Q. Yu) and the China Ministry of Science and Technology Key New Drug Creation and Manufacturing Program (No. 2014ZX9102001002 to JY. Yuan).

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Author notes

  1. Hui Huang and Junxing Niu contributed equally to the paper.

Authors and Affiliations

  1. Division of Tumor Pharmacology, Shanghai Institute of Materia Medica, Chinese Academy of Sciences, 555 Zu Chong Zhi Road, Zhangjiang Hi-Tech Park, Shanghai, 201203, China

    Hui Huang, Junxing Niu, Fei Wang & Qiang Yu

  2. University of Chinese Academy of Sciences, No. 19A Yuquan Road, Beijing, 100049, China

    Hui Huang, Junxing Niu, Fei Wang & Qiang Yu

  3. Stake Key Laboratory Cultivation Base for TCM Quality and Efficacy, School of Pharmacy, Nanjing University of Chinese Medicine, 138 Xianlin Avenue, Nanjing, 210046, Jiangsu Province, China

    Lihong Hu

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Huang, H., Niu, J., Wang, F. et al. A natural compound derivative P-13 inhibits STAT3 signaling by covalently inhibiting Janus kinase 2. Invest New Drugs 37, 452–460 (2019). https://doi.org/10.1007/s10637-018-0637-2

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