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Novel Caspase-1 inhibitor CZL80 improves neurological function in mice after progressive ischemic stroke within a long therapeutic time-window

Acta Pharmacologica Sinica volume 43, pages 2817–2827 (2022)Cite this article

Abstract

Progressive ischemic stroke (PIS) is featured by progressive neurological dysfunction after ischemia. Ischemia-evoked neuroinflammation is implicated in the progressive brain injury after cerebral ischemia, while Caspase-1, an active component of inflammasome, exaggerates ischemic brain injury. Current Caspase-1 inhibitors are inadequate in safety and druggability. Here, we investigated the efficacy of CZL80, a novel Caspase-1 inhibitor, in mice with PIS. Mice and Caspase-1−/− mice were subjected to photothrombotic (PT)-induced cerebral ischemia. CZL80 (10, 30 mg·kg−1·d−1, i.p.) was administered for one week after PT onset. The transient and the progressive neurological dysfunction (as foot faults in the grid-walking task and forelimb symmetry in the cylinder task) was assessed on Day1 and Day4-7, respectively, after PT onset. Treatment with CZL80 (30 mg/kg) during Day1-7 significantly reduced the progressive, but not the transient neurological dysfunction. Furthermore, we showed that CZL80 administered on Day4-7, when the progressive neurological dysfunction occurred, produced significant beneficial effects against PIS, suggesting an extended therapeutic time-window. CZL80 administration could improve the neurological function even as late as Day43 after PT. In Caspase-1−/− mice with PIS, the beneficial effects of CZL80 were abolished. We found that Caspase-1 was upregulated during Day4-7 after PT and predominantly located in activated microglia, which was coincided with the progressive neurological deficits, and attenuated by CZL80. We showed that CZL80 administration did not reduce the infarct volume, but significantly suppressed microglia activation in the peri-infarct cortex, suggesting the involvement of microglial inflammasome in the pathology of PIS. Taken together, this study demonstrates that Caspase-1 is required for the progressive neurological dysfunction in PIS. CZL80 is a promising drug to promote the neurological recovery in PIS by inhibiting Caspase-1 within a long therapeutic time-window.

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Fig. 1: CZL80 rescues progressive motor dysfunction in photothrombotic mice.
Fig. 2: Treatment with CZL80 during Day4-7, but not Day1-4 after photothrombosis ameliorates progressive neurological dysfunction.
Fig. 3: CZL80 attenuates the long-term motor function recovery after photothrombotic stroke.
Fig. 4: Caspase-1−/− abolishes the therapeutic effect of CZL80.
Fig. 5: CZL80 cannot inhibit neuronal death in PT-induced progressive stroke.
Fig. 6: Microglia activation in the progressive stroke.
Fig. 7: CZL80 inhibits the activation of microglia.
Fig. 8: Angiogenesis and improved blood flow have limited contribution to the pharmacological effects of CZL80.

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Acknowledgements

This work was funded by the National Natural Science Foundation of China (82173792 and 81822044), Zhejiang Provincial Natural Science Foundation (LZ21H310001) and the Starry Night Science Fund of Zhejiang University Shanghai Institute for Advanced Study (SN-ZJU-SIAS-0011). We are grateful to the Imaging Facilities, Zhejiang University School of Medicine for the help in microscopy.

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  1. These authors contributed equally: Ling Pan, Wei-dong Tang

Authors and Affiliations

  1. Institute of Pharmacology and Toxicology, College of Pharmaceutical Sciences, Zhejiang University, Hangzhou, 310058, China

    Ling Pan, Wei-dong Tang, Ke Wang, Qi-feng Fang, Meng-ru Liu, Zhan-xun Wu, Yi Wang, Wei-wei Hu, Zhong Chen & Xiang-nan Zhang

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

    Yi Wang & Zhong Chen

  3. Department of Pharmachemistry, College of Pharmaceutical Sciences, Zhejiang University, Hangzhou, 310058, China

    Sun-liang Cui & Ting-jun Hou

  4. Department of Pharmacology, School of Medicine and Life Sciences, Nanjing University of Chinese Medicine, Nanjing, 210029, China

    Gang Hu

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Contributions

XNZ and ZC designed research. LP and WDT planned and performed the experiments, analyzed the data and edited the paper. KW, QFF, MRL and ZXW participated in the biochemistry experiments. YW, SLC, GH, TJH and WWH participated in the project discussion and analyzed the data. LP contributed to writing the paper, and XNZ reviewed/edited the paper.

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Correspondence to Zhong Chen or Xiang-nan Zhang.

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Pan, L., Tang, Wd., Wang, K. et al. Novel Caspase-1 inhibitor CZL80 improves neurological function in mice after progressive ischemic stroke within a long therapeutic time-window. Acta Pharmacol Sin 43, 2817–2827 (2022). https://doi.org/10.1038/s41401-022-00913-7

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  • DOI: https://doi.org/10.1038/s41401-022-00913-7

Keywords

  • progressive ischemic stroke
  • neuroinflammation
  • Caspase-1 inhibitor
  • microglia
  • neurological dysfunction
  • therapeutic time-window

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