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An Inhibitor of Macrophage Arginine Transport and Nitric Oxide Production (CNI-1493) Prevents Acute Inflammation and Endotoxin Lethality

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Abstract

Background

Nitric oxide (NO), a small effector molecule produced enzymatically from L-arginine by nitric oxide synthase (NOS), is a mediator not only of important homeostatic mechanisms (e.g., blood vessel tone and tissue perfusion), but also of key aspects of local and systemic inflammatory responses. Previous efforts to develop inhibitors of NOS to protect against NO-mediated tissue damage in endotoxin shock have been unsuccessful, largely because such competitive NOS antagonists interfere with critical vasoregulatory NO production in blood vessels and decrease survival in endotoxemic animals. Accordingly, we sought to develop a pharmaceutical approach to selectively inhibit NO production in macrophages while sparing NO responses in blood vessels.

Materials and Methods

The processes of cytokine-inducible L-arginine transport and NO production were studied in the murine macrophage-like cell line (RAW 264.7). A series of multivalent guanylhydrazones were synthesized to inhibit cytokine-inducible L-arginine transport. One such compound (CNI-1493) was studied further in animal models of endothelial-derived relaxing factor (EDRF) activity, carrageenan inflammation, and lethal lipopolysaccharide (LPS) challenge.

Results

Upon activation with cytokines, macrophages increase transport of L-arginine to support the production of NO by NOS. Since endothelial cells do not require this additional arginine transport to produce NO, we reasoned that a competitive inhibitor of cytokine-inducible L-arginine transport would not inhibit EDRF activity in blood vessels, and thus might be effectively employed against endotoxic shock. CNI-1493, a tetravalent guanylhydrazone, proved to be a selective inhibitor of cytokine-inducible arginine transport and NO production, but did not inhibit EDRF activity. In mice, CNI-1493 prevented the development of carrageenan-induced footpad inflammation, and conferred protection against lethal LPS challenge.

Conclusions

A selective inhibitor of cytokine-inducible L-arginine transport that does not inhibit vascular EDRF responses is effective against endotoxin lethality and significantly reduces inflammatory responses.

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Acknowledgments

This work was supported in part by National Institutes of Health Grant R01DK49283 (KT), a Faculty Fellowship Award from the American College of Surgeons (KT), a grant from Cytokine Network Inc., and Institutional Funding from the Picower Institute.

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Authors and Affiliations

  1. The Picower Institute for Medical Research, Manhasset, New York, 11030, USA

    Marina Bianchi, Peter Ulrich, Malcolm Meistrell III, Gary A. Zimmerman, Helena Schmidtmayerova, Michael Bukrinsky, Thomas Donnelley, Richard Bucala, Barbara Sherry, Kirk R. Manogue, Anthony Cerami & Kevin J. Tracey

  2. New York Hospital-Cornell University Medical College, New York, New York, USA

    Gary A. Zimmerman

  3. Laboratory of Biomedical Science, Department of Surgery, North Shore University Hospital-Cornell University Medical College, Manhasset, New York, USA

    Ona Bloom, Anthony J. Tortolani & Kevin J. Tracey

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  1. Marina Bianchi
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Bianchi, M., Ulrich, P., Bloom, O. et al. An Inhibitor of Macrophage Arginine Transport and Nitric Oxide Production (CNI-1493) Prevents Acute Inflammation and Endotoxin Lethality. Mol Med 1, 254–266 (1995). https://doi.org/10.1007/BF03401550

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