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Targeted delivery of nitric oxide via a ‘bump-and-hole’-based enzyme–prodrug pair

Nature Chemical Biology volume 15, pages 151–160 (2019)Cite this article

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Abstract

The spatiotemporal generation of nitric oxide (NO), a versatile endogenous messenger, is precisely controlled. Despite its therapeutic potential for a wide range of diseases, NO-based therapies are limited clinically due to a lack of effective strategies for precisely delivering NO to a specific site. In the present study, we developed a novel NO delivery system via modification of an enzyme–prodrug pair of galactosidase–galactosyl-NONOate using a ‘bump-and-hole’ strategy. Precise delivery to targeted tissues was clearly demonstrated by an in vivo near-infrared imaging assay. The therapeutic potential was evaluated in both rat hindlimb ischemia and mouse acute kidney injury models. Targeted delivery of NO clearly enhanced its therapeutic efficacy in tissue repair and function recovery and abolished side effects due to the systemic release of NO. The developed protocol holds broad applicability in the targeted delivery of important gaseous signaling molecules and offers a potent tool for the investigation of relevant molecular mechanisms.

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Fig. 1: ‘Bump-and-hole’-based prodrug design.
Fig. 2: Engineering β-galactosidases for selectively activating ‘bumpy’ glycosyl-probes.
Fig. 3: Synthesis and evaluation of MeGal-NO prodrug.
Fig. 4: Targeted delivery of nitric oxide enhances the angiogenesis and blood perfusion, modulates inflammatory response, and ameliorates muscle recovery in a rat hindlimb ischemia model.
Fig. 5: Targeted delivery of nitric oxide attenuates renal injury and promotes kidney repair in mouse acute kidney injury (AKI) model.
Fig. 6: Targeted delivery of nitric oxide rescues the vulnerability of eNOS-deficient mice to acute kidney injury (AKI).

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Data availability

The data that support the findings of this study are available within the article and supplementary information files and from the corresponding authors upon reasonable request.

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Acknowledgements

We thank J. Xiao at Shanghai University for kindly donating eNOS-knockout mice. This study was supported by National Natural Science Foundation of China (Nos. 81522023 to Q.Z., 81830060 to D.K., 81603064 to J.H., 81871500 to Q.Z., and 91639113 to Q.Z.) and National Key R&D Program of China (2017YFC1103501 to D.K. and Q.Z.).

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Author notes

  1. These authors contributed equally: Jingli Hou, Yiwa Pan, Dashuai Zhu.

Authors and Affiliations

  1. State Key Laboratory of Medicinal Chemical Biology and College of Pharmacy, Nankai University, Tianjin, China

    Jingli Hou, Yueyuan Fan, Jiansong Cheng & Jie Shen

  2. Tianjin Key Laboratory on Technologies Enabling Development of Clinical Therapeutics and Diagnostics, School of Pharmacy, Tianjin Medical University, Tianjin, PR China

    Jingli Hou & Yangping Liu

  3. State key Laboratory of Medicinal Chemical Biology, Key Laboratory of Bioactive Materials, Ministry of Education, and College of Life Science, Nankai University, Tianjin, China

    Yiwa Pan, Yongzhen Wei, He Wang, Kang Qin, Deling Kong & Qiang Zhao

  4. School of Medicine, Nankai University, Tianjin, China

    Dashuai Zhu & Yongzhe Che

  5. Department of Genitourinary Oncology, Tianjin Medical University Cancer Institute and Hospital, National Clinical Research Center for Cancer, Key Laboratory of Cancer Prevention and Therapy, Tianjin, China

    Guowei Feng

  6. Center for Research and Development of Chinese Medicine, Tianjin State Key Laboratory of Modern Chinese Medicine, Tianjin University of Traditional Chinese Medicine, Tianjin, China

    Tiechan Zhao & Yan Zhu

  7. Spine Department, Tianjin Hospital, Tianjin, China

    Qiang Yang

  8. Department of Chemistry and Center for Therapeutics and Diagnostics, Georgia State University, Atlanta, GA, USA

    Peng George Wang

  9. Jiangsu Center for the Collaboration and Innovation of Cancer Biotherapy, Cancer Institute, Xuzhou Medical University, Xuzhou, Jiangsu, China

    Qiang Zhao

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Contributions

J.S. and J.C. conceived the original concept and initiated this project. Q.Z. designed the experiment and supervised the entire project. Y.F. performed the expression of enzymes under the supervision of J.C. J.H. carried out the synthesis of all compounds and fluorescent probes. G.F. established ischemia animal models. Y.P. and D.Z. performed the in vivo evaluation and analyzed data under the supervision of Q.Z. J.H. and H.W. performed the in vitro releasing test of NO. Y.W. carried out the RT-PCR assay. T.Z. performed micro-CT analysis under the supervision of Y.Z. Y.L. helped with EPR analysis. K.Q., Y.C., and Q.Y. helped with data collection. P.G.W. and D.K. provided critical feedback and helped in review of the article. Q.Z., J.C., and J.H. wrote the paper with input from other authors.

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Correspondence to Jiansong Cheng, Jie Shen or Qiang Zhao.

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Hou, J., Pan, Y., Zhu, D. et al. Targeted delivery of nitric oxide via a ‘bump-and-hole’-based enzyme–prodrug pair. Nat Chem Biol 15, 151–160 (2019). https://doi.org/10.1038/s41589-018-0190-5

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