Selective iNOS inhibitor, ONO1714 successfully retards the development of high-cholesterol diet induced atherosclerosis by novel mechanism



      We have reported that inducible nitric oxide synthase (iNOS) is present only in deep areas of plaque in atherosclerosis. However, the role of iNOS in the development of atherosclerosis is not well known. We therefore investigated the relevance of iNOS inhibition.

      Methods and results

      Seven groups of male rabbits were fed a 0.5% high-cholesterol diet (HCD) for 8 weeks. Gp1-HCD was fed HCD only; Gp2-O17 was fed HCD with ONO1714, an iNOS inhibitor; Gp3-AG was fed HCD with amino-guanidine (AG), an iNOS inhibitor; Gp4-AR was fed HCD with l-arginine; Gp5-AR-O17 was fed HCD with l-arginine with ONO1714; Gp6-LNA was fed HCD with l-NAME (a NOS inhibitor); and Gp7-LN-O17 was fed HCD with l-NAME plus ONO1714. ONO1714 decreased atherosclerosis by about 70% (area occupied by lesions: 3.0 ± 0.4% in Gp2-O17 versus 10.3 ± 1.6% in Gp1-HCD) and also decreased atherosclerosis in Gp7-LN-O17. The ONO compound enhanced the atheroprotective effect of l-arginine. Amino-guanidine also showed an anti-atherosclerotic effect. Tone-related basal NO release and acetylcholine-induced NO-dependent relaxation were improved in Gp2-O17 and Gp5-AR-O17. O2 release was decreased in Gp2-O17 and Gp7-LN-O17.


      ONO1714 retards the progression of atherosclerosis in rabbits. Although the up-regulation of endothelial nitric oxide synthase (eNOS) and the decrease of O2 may play roles in this retardation, the inhibition of iNOS may be the principal factor, alone was not sufficient.


      ACh (acetylcholine chloride), EDR (endothelium-dependent relaxation), eNOS (endothelial nitric oxide synthase), iNOS (inducible nitric oxide synthase), MCLA (2-methyl-6-(p-methoxyphenyl)-3,7-dihydroimidazo 1,2-alphapyrazin-3-one), l-NAME (nitroarginine methylester), l-NMA (NG-monomethyl-l-arginine acetate, inhibitor of NO synthase), NO (nitric oxide), NOS (nitric oxide synthase), NTG (nitroglycerine), PGF2α (prostaglandin F2α)


      To read this article in full you will need to make a payment

      Purchase one-time access:

      Academic & Personal: 24 hour online accessCorporate R&D Professionals: 24 hour online access
      One-time access price info
      • For academic or personal research use, select 'Academic and Personal'
      • For corporate R&D use, select 'Corporate R&D Professionals'


      Subscribe to Atherosclerosis
      Already a print subscriber? Claim online access
      Already an online subscriber? Sign in
      Institutional Access: Sign in to ScienceDirect


        • Hayashi T.
        • Kuzuya M.
        • Funaki C.
        • Kuzuya F.
        β-Very low density lipoprotein inhibits endothelium-dependent relaxation in atherosclerotic vessels.
        Blood Vessels. 1989; 26: 290-299
        • Chester A.H.
        • O’Neil G.S.
        • Moncada S.
        • Tadjkarimi S.
        • Yacoub M.H.
        Low basal and stimulated release of nitric oxide in atherosclerotic epicardial coronary arteries.
        Lancet. 1990; 336: 897-900
        • Cayatte A.J.
        • Palacino J.J.
        • Horten K.
        • Cohen R.A.
        Chronic inhibition of nitric oxide production accelerates neointima formation and impairs endothelial function in hypercholesterolemic rabbits.
        Arterioscler Thromb Vasc Biol. 1994; 14: 746-752
        • Ignarro L.J.
        Biological actions and properties of endothelium-derived nitric oxide formed and released from artery and vein.
        Circ Res. 1989; 65: 1-21
        • Harrison D.G.
        • Armstrong M.L.
        • Freiman P.C.
        • Heistad D.D.
        Restoration of endothelium-dependent relaxation by dietary treatment of atherosclerosis.
        J Clin Invest. 1987; 80: 1808-1811
        • Hayashi T.
        • Esaki T.
        • Muto E.
        • Kano H.
        • Iguchi A.
        Endothelium-dependent relaxation of rabbit atherosclerotic aorta was not restored by control of hyperlipidemia—the possible role of peroxynitrite.
        Atherosclerosis. 1999; 147: 349-367
        • Shears 2nd, L.L.
        • Kibbe M.R.
        • Murdock A.D.
        • et al.
        Efficient inhibition of intimal hyperplasia by adenovirus-mediated inducible nitric oxide synthase gene transfer to rats and pigs in vivo.
        J Am Coll Surg. 1998; 187: 295-306
        • Gunnett C.A.
        • Lund D.D.
        • Chu Y.
        • et al.
        NO-dependent vasorelaxation is impaired after gene transfer of inducible NO-synthase.
        Arterioscler Thromb Vasc Biol. 2001; 21: 1281-1287
        • Naka M.
        • Nanbu T.
        • Kobayashi K.
        • et al.
        A potent inhibitor of inducible nitric oxide synthase, ONO-1714, a cyclic amidine derivative.
        Biochem Biophys Res Commun. 2000; 270: 663-667
        • Ogawa D.
        • Shikata K.
        • Matsuda M.
        • et al.
        Protective effect of a novel and selective inhibitor of inducible nitric oxide synthase on experimental crescentic glomerulonephritis in WKY rats.
        Nephrol Dial Transplant. 2002; 17: 2117-2121
      1. Lipid Research Clinics Program Manual of Laboratory Operations, vol. 1, second ed., Lipid and Lipoprotein Analysis. U.S. Dept. of Health, Education and Welfare, Publ. no. (NIH). Washington, DC, U.S. Govt. Printing Office. 1982; 76–628.

        • Hayashi T.
        • Fukuto J.M.
        • Ignarro L.J.
        • Chaudhuri G.
        Basal release of nitric oxide from aortic rings are greater in female rabbits than in male rabbits: implications for atherosclerosis.
        Proc Natl Acad Sci USA. 1992; 89: 11259-11263
        • Weiner B.H.
        • Ockene I.S.
        • Hoogasian J.J.
        Inhibition of atherosclerosis by cod-liveroil in a hyperlipidemic swine model.
        N Engl J Med. 1986; 315: 841-845
        • Esaki T.
        • Hayashi T.
        • Muto E.
        • et al.
        Expression of inducible nitric oxide synthase in T lymphocytes and macrophages of cholesterol-fed rabbits.
        Atherosclerosis. 1997; 128: 39-46
        • Hayashi T.
        • Esaki T.
        • Muto E.
        • et al.
        Endothelium-dependent relaxation of rabbit atherosclerotic aorta was not restored by control of hyperlipidemia—the possible role of peroxynitrite.
        Atherosclerosis. 1999; 147: 349-367
        • Chilvers E.R.
        • Giembycz M.A.
        • Challiss R.A.
        • Barnes B.J.
        • Nahorski S.R.
        Lack of effect of zaprinast on methacholine-induced contraction and inositol 1,4,5-trisphosphate accumulation in bovine tracheal smooth muscle.
        Br J Pharmacol. 1991; 103: 1119-1125
        • Matsuda N.
        • Hattori Y.
        • Zhang X.-H.
        • et al.
        Contractions to histamine in pulmonary and mesenteric arteries from endotoxemic rabbits: modulation by vascular expressions of inducible nitric-oxide synthase and histamine H1-receptors.
        J Pharmacol Exp Ther. 2003; 307: 175-181
        • Kano H.
        • Hayashi T.
        • Sumi D.
        • et al.
        A HMG-CoA reductase inhibitor improved regression of atherosclerosis without affecting serum lipid levels—relevance of upregulation of eNOS.
        Biochem Biophys Res Commun. 1999; 259: 414-419
        • Kumar N.T.
        • Hayashi T.
        • Sumi D.
        • Kano H.
        • Iguchi A.
        Stabilization of atherosclerosis by a HMG-CoA reductase inhibitor—effects of increasing basal NO and decreasing superoxide.
        Am J Physiol Heart Circ Physiol. 2001; 281: H75-H83
        • Esaki T.
        • Hayashi T.
        • Iguchi A.
        Expression of inducible nitric oxide synthase and Fas/Fas ligand correlates with the incidence of apoptotic cell death in atheromatous plaques of human coronary arteries.
        Nitric Oxide. 2000; 4: 561-571
        • Kuhlencordt P.J.
        • Chen J.
        • Han F.
        • Astern J.
        • Huang P.L.
        Genetic deficiency of inducible nitric oxide synthase reduces atherosclerosis and lowers plasma lipid peroxides in apolipoprotein E-knockout mice.
        Circulation. 2001; 103: 3099-3104
        • Ihrig M.
        • Dangler C.A.
        • Fox J.G.
        Mice lacking inducible nitric oxide synthase develop spontaneous hypercholesterolemia and aortic atheromas.
        Atherosclerosis. 2001; 156: 103-107
        • Niu X.L.
        • Yang X.
        • Hoshiai K.
        • et al.
        Inducible nitric oxide synthase deficiency does not affect the susceptibility of mice to atherosclerosis but increases collagen content in lesions.
        Circulation. 2001; 103: 1115-1120
        • Ungvari Z.
        • Csiszar A.
        • Edwards J.G.
        • et al.
        Increased superoxide production in coronary arteries in hyperhomocysteinemia: role of tumor necrosis factor-alpha, NAD(P)H oxidase, and inducible nitric oxide synthase.
        Arterioscler Thromb Vasc Biol. 2003; 23: 418-424
        • Cooke C.L.
        • Davidge S.T.
        Peroxynitrite increases iNOS through NF-kappaB and decreases prostacyclin synthase in endothelial cells.
        Am J Physiol Cell Physiol. 2002; 282: C395-C402
        • Chen J.
        • Kuhlencordt P.
        • Urano F.
        • et al.
        Effects of chronic treatment with l-arginine on atherosclerosis in apoE knockout and apoE/inducible NO synthase double-knockout mice.
        Arterioscler Thromb Vasc Biol. 2003; 23: 97-103
        • Simonet S.
        • Rupin A.
        • Badier-Commander C.
        • et al.
        Evidence for superoxide anion generation in aortas of cholesterol-fed rabbits treated with l-arginine.
        Eur J Pharmacol. 2004; 492: 211-216
        • Yoon S.K.
        • Lee K.N.
        • Lee J.H.
        • Jeong J.S.
        • Kwak J.-Y.
        Pulmonary vascular dilation induced by l-arginine: correlation with induction of endothelial nitric oxide synthase in a rabbit model.
        Acta Radiol. 2005; 46: 48-54
        • Hayashi T.
        • Iguchi A.
        • Ignarro L.J.
        Gene transfer of endothelial NO synthase (eNOS), but not eNOS plus inducible NOS regressed atherosclerosis in rabbits.
        Cardiovasc Res. 2004; 61: 339-351