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Treatment of cerebral ischemia by disrupting ischemia-induced interaction of nNOS with PSD-95

Nature Medicine volume 16, pages 1439–1443 (2010)Cite this article

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A Corrigendum to this article was published on 07 September 2011

This article has been updated

Abstract

Stroke is a major public health problem leading to high rates of death and disability in adults1,2. Excessive stimulation of N-methyl-D-aspartate receptors (NMDARs) and the resulting neuronal nitric oxide synthase (nNOS) activation are crucial for neuronal injury after stroke insult3,4,5,6,7. However, directly inhibiting NMDARs or nNOS can cause severe side effects because they have key physiological functions in the CNS5,8,9,10,11,12. Here we show that cerebral ischemia induces the interaction of nNOS with postsynaptic density protein-95 (PSD-95). Disrupting nNOS–PSD-95 interaction via overexpressing the N-terminal amino acid residues 1–133 of nNOS (nNOS-N1–133) prevented glutamate-induced excitotoxicity and cerebral ischemic damage. Given the mechanism of nNOS–PSD-95 interaction, we developed a series of compounds and discovered a small-molecular inhibitor of the nNOS-PSD-95 interaction, ZL006. This drug blocked the ischemia-induced nNOS–PSD-95 association selectively, had potent neuroprotective activity in vitro and ameliorated focal cerebral ischemic damage in mice and rats subjected to middle cerebral artery occlusion (MCAO) and reperfusion. Moreover, it readily crossed the blood-brain barrier, did not inhibit NMDAR function, catalytic activity of nNOS or spatial memory, and had no effect on aggressive behaviors. Thus, this new drug may serve as a treatment for stroke, perhaps without major side effects.

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Figure 1: Glutamate-induced nNOS translocation from cytosol to plasma membrane via its binding to PSD-95.
Figure 2: Dissociation of PSD-95–nNOS prevents excitotoxicity and cerebral ischemia.
Figure 3: Effects of target compounds on nNOS–PSD-95 interaction.
Figure 4: ZL006 prevents NMDAR-dependent excitotoxicity and cerebral ischemia.

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Change history

  • 07 September 2011

     In the version of this article initially published, one of the structural details for the compound ZL010 in Figure 3f was incorrect. The R1 of ZL010 should be OCH3, not OH. This error does not affect the interpretation of the data or the conclusions of the paper. The error has been corrected in the HTML and PDF versions of the article.

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Acknowledgements

This work was supported by grants from the National Natural Science Foundation of China ((30971021, 81030023) (D.-Y.Z.) and (30901550) (C.-X.L.)). We thank X. Jin, Q.-P. Li, C.-C. Cao, H.-H. Zhou, W.-X. Sun, D.-L. Wu, Y. Hu and J. Zhang for technical assistance.

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Author notes

  1. Li Zhou, Fei Li and Hai-Bing Xu: These authors contributed equally to this work.

Authors and Affiliations

  1. Department of Pharmacology, School of Pharmacy, Nanjing Medical University, Nanjing, China

    Li Zhou, Hai-Bing Xu, Chun-Xia Luo, Hai-Yin Wu, Ming-Mei Zhu, Qi-Gang Zhou & Dong-Ya Zhu

  2. Department of Medicinal Chemistry, School of Pharmacy, Nanjing Medical University, Nanjing, China

    Fei Li & Xing Ji

  3. Laboratory of Cerebrovascular Disease, Nanjing Medical University, Nanjing, China

    Chun-Xia Luo, Hai-Yin Wu, Qi-Gang Zhou & Dong-Ya Zhu

  4. Department of Neurobiology, Nanjing Medical University, Nanjing, China

    Wei Lu

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Contributions

L.Z. contributed to the design of the study and performed the cell culture studies, coimmunoprecipitation, western blotting, imaging of NO synthesis and surgical preparation. F.L. performed the design and synthesis of target compounds. H.-B.X. contributed to coimmunoprecipitation, structure-activity relationship analyses and behavioral analyses. C.-X.L. contributed to the design of the study and performed cell culture and morphological analysis of cortical neurons. H.-Y.W. participated in surgical preparation, infarct volume measurement, neuroscore assessment, physiological parameter determination, platelet aggregation and bleeding time measurements. M.-M.Z. performed behavioral analyses, lentivirus production and stereotaxic injection. W.L. performed electrophysiological experiments. X.J. participated in synthesis of target compounds. Q.-G.Z. participated in western blotting analysis. D.-Y.Z. initiated the project and participated in the design of the studies. All authors contributed to data analysis.

Corresponding authors

Correspondence to Chun-Xia Luo or Dong-Ya Zhu.

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The authors declare no competing financial interests.

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Supplementary Figures 1–14, Supplementary Tables 1 and 2 and Supplementary Methods (PDF 845 kb)

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Zhou, L., Li, F., Xu, HB. et al. Treatment of cerebral ischemia by disrupting ischemia-induced interaction of nNOS with PSD-95. Nat Med 16, 1439–1443 (2010). https://doi.org/10.1038/nm.2245

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