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Inflachromene attenuates seizure severity in mouse epilepsy models via inhibiting HMGB1 translocation

Acta Pharmacologica Sinica volume 44, pages 1737–1747 (2023)Cite this article

Abstract

Epilepsy is not well controlled by current anti-seizure drugs (ASDs). High mobility group box 1 (HMGB1) is a DNA-binding protein in the nucleus regulating transcriptional activity and maintaining chromatin structure and DNA repair. In epileptic brains, HMGB1 is released by activated glia and neurons, interacting with various receptors like Toll-like receptor 4 (TLR4) and downstream glutamatergic NMDA receptor, thus enhancing neural excitability. But there is a lack of small-molecule drugs targeting the HMGB1-related pathways. In this study we evaluated the therapeutic potential of inflachromene (ICM), an HMGB-targeting small-molecule inhibitor, in mouse epilepsy models. Pentylenetetrazol-, kainic acid- and kindling-induced epilepsy models were established in mice. The mice were pre-treated with ICM (3, 10 mg/kg, i.p.). We showed that ICM pretreatment significantly reduced the severity of epileptic seizures in all the three epilepsy models. ICM (10 mg/kg) exerted the most apparent anti-seizure effect in kainic acid-induced epileptic status (SE) model. By immunohistochemical analysis of brain sections from kainic acid-induced SE mice, we found that kainic acid greatly enhanced HMGB1 translocation in the hippocampus, which was attenuated by ICM pretreatment in subregion- and cell type-dependent manners. Notably, in CA1 region, the seizure focus, ICM pretreatment mainly inhibited HMGB1 translocation in microglia. Furthermore, the anti-seizure effect of ICM was related to HMGB1 targeting, as pre-injection of anti-HMGB1 monoclonal antibody (5 mg/kg, i.p.) blocked the seizure-suppressing effect of ICM in kainic acid-induced SE model. In addition, ICM pretreatment significantly alleviated pyramidal neuronal loss and granule cell dispersion in kainic acid-induced SE model. These results demonstrate that ICM is an HMGB-targeting small molecule with anti-seizure potential, which may help develop a potential drug for treating epilepsy.

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Fig. 1: ICM alleviates seizure severity in PTZ-induced seizure model.
Fig. 2: ICM alleviates seizure severity in hippocampal-kindled seizure model.
Fig. 3: ICM alleviates seizure severity in KA-induced SE model.
Fig. 4: ICM attenuates the increased HMGB1 translocation in KA-induced SE model.
Fig. 5: ICM attenuates the increased translocation of HMGB1 with cell-type specificity in different subregions of hippocampus.
Fig. 6: Anti-HMGB1 mAb abolishes the anti-seizure effect of ICM in KA-induced SE model.
Fig. 7: ICM alleviates pyramidal neuronal loss and granule cell dispersion after KA-induced SE.

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Acknowledgements

This project was supported by grants from the National Key R&D program of China (2021ZD0202803 and 2020YFA0803902), the National Natural Science Foundation of China (82022071), and the Natural Science Foundation of Zhejiang Province (LD22H310003).

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  1. These authors contributed equally: Si-jie Dai, Yu-ying Shao

Authors and Affiliations

  1. Institute of Pharmacology & Toxicology, College of Pharmaceutical Sciences, School of Medicine, Zhejiang University, Hangzhou, 310058, China

    Si-jie Dai, Yu-ying Shao, Jin-yi Sun, Zhi-sheng Li, Jia-ying Shi, Yi Wang & Zhong Chen

  2. Key Laboratory of Neuropharmacology and Translational Medicine of Zhejiang Province, School of Pharmaceutical Sciences, Zhejiang Chinese Medical University, Hangzhou, 310053, China

    Yang Zheng, Meng-qi Yan, Xiao-yun Qiu, Ceng-lin Xu, Yi Wang & Zhong Chen

  3. CRI Center for Chemical Proteomics, Department of Chemistry, Seoul National University, Seoul, 08826, Republic of Korea

    Wan-sang Cho, Sihyeong Yi & Seung Bum Park

  4. Department of Pharmacology, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences, Okayama, Japan

    Masahiro Nishibori

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Contributions

ZC, YW, and SBP designed the research. SJD, YYS, MQY, and XYQ conducted the experiments. SJD, YZ, and YW conducted the data analysis. YZ, JYSun, ZSL, JYShi, CLX, WSC, SY, SBP, MN, YW, and ZC provided technical guidance and contributed to the data discussion. SJD, YYS, YZ, and YW wrote the manuscript. YW and ZC supervised all aspects of the work.

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Correspondence to Yi Wang or Zhong Chen.

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Dai, Sj., Shao, Yy., Zheng, Y. et al. Inflachromene attenuates seizure severity in mouse epilepsy models via inhibiting HMGB1 translocation. Acta Pharmacol Sin 44, 1737–1747 (2023). https://doi.org/10.1038/s41401-023-01087-6

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