Nitric oxide (NO) is a widespread, potent, biological mediator that has many physiological and pathophysiological roles1. Research in the field of NO appears to have followed a straightforward path, and the findings have been progressive: NO and cyclic GMP are involved in vasodilatation; glycerol trinitrate relaxes vascular smooth muscles by bioconversion to NO; mammalian cells synthesize NO; and last, NO mediates vasodilatation by stimulating the soluble guanylate cyclase (sGC), a heterodimeric (α/β) haem protein that converts GTP to cGMP2–4. Here we report the discovery of a regulatory site on sGC. Using photoaffinity labelling, we have identified the cysteine 238 and cysteine 243 region in the α1-subunit of sGC as the target for a new type of sGC stimulator. Moreover, we present a pyrazolopyridine, BAY 41-2272, that potently stimulates sGC through this site by a mechanism that is independent of NO. This results in antiplatelet activity, a strong decrease in blood pressure and an increase in survival in a low-NO rat model of hypertension, and as such may offer an approach for treating cardiovascular diseases.
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Moncada, S., Palmer, R. M. & Higgs, E. A. Nitric oxide: physiology, pathophysiology, and pharmacology. Pharmacol. Rev. 43, 109–142 (1991).
Furchgott, R. F. Endothelium-derived relaxing factor: Discovery, early studies, and identification as nitric oxide (Nobel lecture). Angew. Chem. Int. Ed. Engl. 38, 1870–1880 (1999).
Murad, F. Discovery of some of the biological effects of nitric oxide and its role in cell signaling (Nobel lecture). Angew. Chem. Int. Ed. Engl. 38, 1857–1868 (1999).
Ignarro, L. J. Nitric oxide: A unique endogenous signaling molecule in vascular biology (Nobel lecture). Angew. Chem. Int. Ed. Engl. 38, 1882–1892 (1999).
Straub, A. et al. Substituted pyrazole derivatives. WO 2,000,006,568 (2000). Chem. Abstr. 132, 122629 (2000)
Garthwaite, J. et al. Potent and selective inhibition of nitric oxide-sensitive guanylyl cyclase by 1H[1,2,4]oxadiazolo[4,3-a]quinoxalin-1-one. Mol. Pharmacol. 48, 184–188 (1995).
Schrammel, A., Behrends, S., Schmidt, K., Koesling, D. & Mayer, B. Characterization of 1H[1,2,4]oxadiazolo[4,3-a]quinoxalin-1-one as a heme-site inhibitor of nitric oxide-sensitive guanylyl cyclase. Mol. Pharmacol. 50, 1–5 (1996).
Brunner, F., Schmidt, K., Nielsen, E. B. & Mayer, B. Novel guanylyl cyclase inhibitor potently inhibits cyclic GMP accumulation in endothelial cells and relaxation of bovine pulmonary artery. J. Pharmacol. Exp. Ther. 277, 48–53 (1996).
Friebe, A., Schultz G. & Koesling, D. Sensitizing soluble guanylate cyclase to become a highly CO-sensitive enzyme. EMBO J. 15, 6863–6868 (1996).
Hoenicka, M. et al. Purified soluble guanylyl cyclase expressed in a baculovirus/Sf9 system: stimulation by YC-1, NO and CO. J. Mol. Med. 77, 14–23 (1999).
Nakane, M. et al. Molecular cloning and expression of cDNAs coding for soluble guanylate cyclase from rat lung. J. Biol. Chem. 265, 16841–16845 (1990).
Zabel, U., Weeger, M., La, M. & Schmidt, H. H. H. W. Human soluble guanylate cyclase: functional expression and revised isoenzyme family. Biochem. J. 335, 51–57 (1998).
Stasch, J. P., Kazda, S. & Neuser, D. Different effects of ANP and nitroprusside on cyclic GMP extrusion of isolated aorta. Eur. J. Pharmacol. 174, 279–282 (1989).
Becker, E. M. et al. The vasodilator-stimulated phosphoprotein (VASP): target of YC-1 and nitric oxide effects in human and rat platelets. J. Cardiovasc. Pharmacol. 35, 390–397 (2000).
Mülsch, A. et al. Effect of YC-1, an NO-independent, superoxide-sensitive stimulator of soluble guanylyl cyclase, on smooth muscle responsiveness to nitrovasodilators. Br. J. Pharmacol. 120, 681–689 (1997).
Ko, F. N., Wu, C. C., Kuo, S. C., Lee, F. Y. & Teng, C. M. YC-1, a novel activator of platelet guanylate cyclase. Blood 84, 4226–4233 (1994).
Wu, C. C., Ko, F. N., Kuo, S. C., Lee, F. Y. & Teng, C. M. YC-1 inhibited human platelet aggregation through NO-independent activation of soluble guanylate cyclase. Br. J. Pharmacol. 116, 1973–1978 (1995).
Becker, E. M. et al. Generation and characterization of a stable soluble guanylate cyclase-overexpressing CHO cell line. Nitric Oxide 3, 55–66 (1999).
Friebe, A. & Koesling, D. Mechanism of YC-1 induced activation of soluble guanylyl cyclase. Mol. Pharmacol. 53, 123–127 (1998).
Hobbs, A. J. Soluble guanylate cyclase: the forgotten sibling. Trends Pharmacol. Sci. 18, 484–491 (1997).
Galle, J. et al. Effects of the soluble guanylyl cyclase activator, YC-1, on vascular tone, cyclic GMP levels and phosphodiesterase activity. Br. J. Pharmacol. 127, 195–203 (1999).
Hirth-Dietrich, C., Stasch, J. P. & Ganten, D. Transgenic rats with an additional renin gene (TGR(mRen2)27) as a sensitive model for the renal effects of NO synthase inhibition. Naunyn-Schmiedeberg's Arch. Pharmacol. 350, R6 (1994).
Zhao, Y., Schelvis, J. P. M., Babcock, G. T. & Marletta, M. A. Identification of histidine 105 in the β1 subunit of soluble guanylate cyclase as the heme proximal ligand. Biochemistry 37, 4502–4509 (1998).
Zhao, Y. & Marletta, M. A. Localization of the heme binding region in soluble guanylate cyclase. Biochemistry 36, 15959–15964 (1997).
Kharitonov, V. G., Sharma, V. S., Magde, D. & Koesling, D. Kinetics and equilibria of soluble guanylate cyclase ligation by CO: effect of YC-1. Biochemistry 38, 10699–10706 (1999).
Born, G. V. R. & Cross, M. J. The aggregation of blood platelets. J. Physiol. (Lond.) 168, 178–185 (1963).
Laemmli, U. K. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 227, 680–685 (1970).
Ahr, H. H. & Steinke, W. Vasodilation and first experience with quantitative studies in whole-body autoradiography during drug development. Xenobiotic Metab. Dispos. 9, 371–378 (1994).
We thank C. Robyr-Fürstner for the synthesis of YC-1; E. Bischoff for performing the PDE assays; and D. Wood for critical comments on the manuscript.
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Stasch, JP., Becker, E., Alonso-Alija, C. et al. NO-independent regulatory site on soluble guanylate cyclase. Nature 410, 212–215 (2001). https://doi.org/10.1038/35065611
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