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Discovery of novel MIF inhibitors that attenuate microglial inflammatory activation by structures-based virtual screening and in vitro bioassays

Acta Pharmacologica Sinica volume 43, pages 1508–1520 (2022)Cite this article

Abstract

Macrophage migration inhibitory factor (MIF) is a pluripotent pro-inflammatory cytokine and is related to acute and chronic inflammatory responses, immune disorders, tumors, and other diseases. In this study, an integrated virtual screening strategy and bioassays were used to search for potent MIF inhibitors. Twelve compounds with better bioactivity than the prototypical MIF-inhibitor ISO-1 (IC50 = 14.41 μM) were identified by an in vitro enzymatic activity assay. Structural analysis revealed that these inhibitors have novel structural scaffolds. Compound 11 was then chosen for further characterization in vitro, and it exhibited marked anti-inflammatory efficacy in LPS-activated BV-2 microglial cells by suppressing the activation of nuclear factor kappa B (NF-κB) and mitogen-activated protein kinases (MAPKs). Our findings suggest that MIF may be involved in the regulation of microglial inflammatory activation and that small-molecule MIF inhibitors may serve as promising therapeutic agents for neuroinflammatory diseases.

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Fig. 1: Workflow of virtual screening.
Fig. 2: Chemical structures of compounds.
Fig. 3: The 3D presentation of the interactions.
Fig. 4: Effects of the compounds on the production of NO, cell viability, protein levels of iNOS, and COX-2 in LPS-primed BV-2 microglial cells.
Fig. 5: Effects of compounds 5, 9, 11, and 12 on the production of NO and the mRNA expression of pro-inflammatory factors in glial cells.
Fig. 6: The anti-inflammatory effects of MIF tautomerase inhibitors were dependent on MIF expression.
Fig. 7: Competitive inhibition pattern and the inhibition of MIF-mediated abrogation of glucocorticoid activity by compound 11.
Fig. 8: Effects of compound 11 on the generation of pro-inflammatory factors in LPS-primed BV-2 microglial cells.
Fig. 9: Compound 11 inhibits LPS-induced BV-2 microglial cells activation via NF-κB and MAPK signaling.
Fig. 10: Neuroprotective effect of compound 11 on CM from a microglia/neuroblastoma co-culture model.

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Acknowledgements

This study was supported by the National Natural Science Foundation of China (81703496, 81973334, 31970909, 21173156, 81803430, 81773702, and 81373382), Natural Science Foundation of Hubei Province (2020CFB543), and Natural Science Foundation of Jiangsu Province (BE2019650) and the Priority Academic Program Development of the Jiangsu Higher Education Institutes (PAPD), National Center for International Research (2017B01012), and Inner Mongolia Natural Science Foundation (2019MS08197).

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  1. These authors contributed equally: Yu Zhang, Lei Xu, Yao Zhang

Authors and Affiliations

  1. Department of Pharmacy, Hubei Clinical Research Center of Parkinson’s disease, Xiangyang Key Laboratory of Movement Disorders, Xiangyang No.1 People’Hospital, Hubei University of Medicine, Xiangyang, 441000, China

    Yu Zhang, Yao Zhang, Jie Pan & Pu-qing Wang

  2. Institute of Bioinformatics and Medical Engineering, Jiangsu University of Technology, Changzhou, 213001, China

    Lei Xu

  3. Jiangsu Key Laboratory of Neuropsychiatric Diseases and College of Pharmaceutical Sciences, Soochow University, Suzhou, 215123, China

    Sheng Tian, Xue-chu Zhen & Long-Tai Zheng

  4. School of Pharmacy, Baotou Medical College, Baotou, 014060, China

    Huan-ting Li & Bo-wen Gao

  5. College of Pharmaceutical Sciences, Zhejiang University, Hangzhou, 310058, China

    Ting-jun Hou

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Contributions

LTZ, XCZ, and TJH designed the research. Yu Zhang, LX, and Yao Zhang designed the experiments. Yu Zhang, LX, and Yao Zhang conducted virtual screening and biological assays. JP, PQW, ST, HTL, and BWG helped to perform part of biological assays. Yu Zhang, LX, Yao Zhang, LTZ, XCZ, and TJH wrote the manuscript.

Corresponding authors

Correspondence to Ting-jun Hou, Xue-chu Zhen or Long-Tai Zheng.

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Zhang, Y., Xu, L., Zhang, Y. et al. Discovery of novel MIF inhibitors that attenuate microglial inflammatory activation by structures-based virtual screening and in vitro bioassays. Acta Pharmacol Sin 43, 1508–1520 (2022). https://doi.org/10.1038/s41401-021-00753-x

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  • DOI: https://doi.org/10.1038/s41401-021-00753-x

Keywords

  • macrophage migration inhibitory factor
  • virtual screening
  • tautomerase assay
  • naive Bayesian classification
  • neuroinflammation

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