Download PDF
Arzneimittelforschung 2008; 58(2): 71-80
DOI: 10.1055/s-0031-1296471
Liver Therapeutics
Editio Cantor Verlag Aulendorf (Germany)

Nitric Oxide-independent Activation of Soluble Guanylate Cyclase by BAY 60-2770 in Experimental Liver Fibrosis

Andreas Knorr
1   Institute of Cardiovascular Research, Bayer HealthCare AG, Wuppertal, Germany
,
Claudia Hirth-Dietrich
1   Institute of Cardiovascular Research, Bayer HealthCare AG, Wuppertal, Germany
,
Cristina Alonso-Alija
2   Institute of Medicinal Chemistry, Bayer HealthCare AG, Wuppertal, Germany
,
Michael Härter
2   Institute of Medicinal Chemistry, Bayer HealthCare AG, Wuppertal, Germany
,
Michael Hahn
2   Institute of Medicinal Chemistry, Bayer HealthCare AG, Wuppertal, Germany
,
Yvonne Keim
1   Institute of Cardiovascular Research, Bayer HealthCare AG, Wuppertal, Germany
,
Frank Wunder
1   Institute of Cardiovascular Research, Bayer HealthCare AG, Wuppertal, Germany
,
Johannes-Peter Stasch
1   Institute of Cardiovascular Research, Bayer HealthCare AG, Wuppertal, Germany
› Author Affiliations
Further Information

Publication History

Publication Date:
15 December 2011 (online)

    Permissions and Reprints

Summary

Liver cirrhosis is a chronic disease with high mortality rate and need for effective pharmacological intervention. The fibrotic remodelling of liver tissue is crucially dependent on hepatic stellate cell activation. Activation of hepatic stellate cells is reduced by an increase in cyclic guanosine monophosphate (cGMP). Stable cGMP analogues also reduce the contractile response of hepatic stellate cells. However, cGMP production is downregulated in the cirrhotic liver due to the reduced activity of the endothelial nitric oxide synthase.

Objective:

Here we report that the novel activator of soluble guanylate cyclase (sGC), BAY 60-2770 (4-({(4-carboxy-butyl)[2-(5-fluoro-2-{[4’-(trifluoromethyl) biphenyl-4-yl]methoxy}phenyl)ethyl]amino}methyl)benzoic acid), which increases the activity of sGC in a nitric oxide-independent manner, attenuates liver fibrosis in two rat models.

Methods:

The compound was studied in the pig serum model and the carbon tetrachloride model. Fibrosis was assessed by estimating the increase in fibrous collagen by micromorphometry of histological sections stained with Sirius Red/Fast Green and by measuring total hepatic collagen.

Results:

BAY 60-2770, on a recombinant sGC reporter cell line, stimulated the luminescence signals with an EC50 value of 5.4 ±1.2 nmol/L. In the presence of [1] [2] [4] oxadiazolo [4,3-a] quinoxalin-1-one (ODQ; 10 µmol/L) the EC50 was shifted to 0.39 ±0.11 nmol/L. In both fibrosis models, once daily oral administration of BAY 60-2770 concomittantly with the fibrotic stimulus prevented 60–75% of fibrosis, the lowest effective dose being 0.1 mg/kg in the pig serum model and 0.3 mg/kg in the carbon tetrachloride model. The treatment was well tolerated by all animals. The doses used were devoid of any significant influence on systemic blood pressure.

Conclusion:

Nitric oxide-independent activation of sGC might be an innovative therapeutic approach for the treatment of liver fibrosis of necro-inflammatory and immunological origin.

Key words

BAY 60-2770, activation of soluble guanylyl cyclase, antifibrotic effect - Guanylate cyclase, nitric oxide-independent activation, soluble - Liver fibrosis, carbon tetrachloride induced, pig serum induced - Liver therapeutics
 

  • References

  • 1 Friedman SL. Stellate cells: a moving target in hepatic fibrogenesis. Hepatology. 2004; 40: 1041-1003
    CrossrefPubMedGoogle Scholar
  • 2 Reynaert H, Thompson MG, Thomas T, Geerts A. Hepatic stellate cells: role in microcirculation and pathophysiology of portal hypertension. Gut. 2002; 50: 571-581
    CrossrefPubMedGoogle Scholar
  • 3 Gäbele E, Brenner D A, Rippe R A. Liver fibrosis: Signals leading to the amplification of the fibrinogenic hepatic stellate cell. Front Biosc. 2003; 8: d69-d77
    CrossrefPubMedGoogle Scholar
  • 4 Corpechot C, Barbu V, Wendum D, Kinnman N, Rey C, Poupon R et al. Hypoxia-induced VEGF and collagen I expressions are associated with angiogenesis and fibrogenesis in experimental cirrhosis. Hepatology. 2002; 35: 1010-1021
    CrossrefPubMedGoogle Scholar
  • 5 Kawada N, Kuroki T, Uoya M, Inue M, Kobayashi K. Smooth muscle alpha-actin expression in rat hepatic stellate cell is regulated by nitric oxide and cGMP production. Biochem. Biophys Res Commun. 1996; 229: 238-242
    CrossrefPubMedGoogle Scholar
  • 6 Tao J, Mallat A, Gallois C, Belmadani S, Mery PF, Nhieu JT et al. Biological effects of C-type natriuretic peptide in human myofibroblastic hepatic stellate cells. J Biol Chem. 1999; 274: 23761-23769
    CrossrefPubMedGoogle Scholar
  • 7 Rockey DC, Chung JJ, McKee CM, Noble PW. Stimulation of inducible nitric oxide synthase in rat liver by hyaluronan fragments. Hepatology. 1998; 27: 86-92
    CrossrefPubMedGoogle Scholar
  • 8 Van de Casteele M, Omasta A, Janssens S, Roskams T, Desmet V, Nevens F et al. In vivo gen transfer of endothelial nitric oxide synthase decreases portal pressure in anaesthetized CCl4 cirrhotic rats. Gut. 2002; 51: 440-445
    CrossrefPubMedGoogle Scholar
  • 9 Loureiro-Silva MR, Iwakiri Y, Abraldes JG, Haq O, Groszmann RJ. Increased phosphodiesterase-5 expression is involved in the decreased vasodilator response to nitric oxide in cirrhotic rat livers. J Hepatol. 2006; 44: 886-893
    CrossrefPubMedGoogle Scholar
  • 10 Stasch JP, Alonso-Alija C, Apeler H, Dembowsky K, Feurer A, Minuth T et al. Pharmacological actions of a novel NO-independent guanylyl cyclase stimulator, BAY 41– 8543 in vitro studies. Br J Pharmacol. 2002; a 135: 333-343
    CrossrefPubMedGoogle Scholar
  • 11 Stasch IP, Dembowsky K, Perzborn E, Stahl E, Schramm M. Cardiovascular actions of a novel NO independent guanylyl cyclase stimulator BAY 41– 8543 in vivo studies. Br J Pharmacol. 2002; b 135: 344-355
    CrossrefPubMedGoogle Scholar
  • 12 Stasch JP, Schmidt P, Alonso-Alija C, Apeler H, Dembowsky K, Haerter M, Heil M et al. NO– and haem-independent activation of soluble guanylyl cyclase: molecular basis and cardiovascular implications of a new pharmacological principle. Br J Pharmacol. 2002; c 136: 773-783
    CrossrefPubMedGoogle Scholar
  • 13 Alonso-Alija C, Heil M, Flubacher D, Naab P, Stasch JP, Wunder F et al. Novel derivatives of dicarboxylic acid having pharmaceuticals properties. Patent WO-119780-A-2001.03.22.2001
    PubMedGoogle Scholar
  • 14 Wunder F, Stasch JP, Hütter J, Alonso-Alija C, Hüser J, Lohrmann E. A cell-based cGMP assay useful for ultra-high-throughput screening and identification of modulators of the NO/cGMP pathway. Anal Biochem. 2005; 339: 104-112
    CrossrefPubMedGoogle Scholar
  • 15 McLean E, McLean A, Sutton P. Instant Cirrhosis. An improved method for producing cirrhosis of the liver in rats by simultaneous administration of CCl4 and phenobarbitone. Br J Exp Pathol. 1969; 50: 502-506
    PubMedGoogle Scholar
  • 16 Bhunchet E, Wake K. Role of mesenchymal cell populations in porcine serum-induced rat liver fibrosis. Hepatology. 1992; 16: 1452-1473
    CrossrefPubMedGoogle Scholar
  • 17 Hoenicka M, Becker EM, Apeler H, Sirichoke T, Schröder H, Gerzer R et al. Purified soluble guanylyl cyclase expressed in a baculovirus/Sf9 system: stimulation by YC-1, nitric oxide, and carbon monoxide. J Mol Med. 1999; 77: 14-23
    CrossrefPubMedGoogle Scholar
  • 18 Prockop DJ, Udenfriend S. A specific method for the analysis of hydroxyproline in tissues and urine. Anal Biochem. 1960; 1: 228-239
    CrossrefPubMedGoogle Scholar
  • 19 Bergmeyer HU, Hørder M, Rej R. Approved recommendation on IFCC methods for the measurement of catalytic concentration of enzymes. Part 3. IFCC method for alanine aminotransferase. J Clin Chem Clin Biochem. 1986; a 24: 481-495
    PubMedGoogle Scholar
  • 20 Bergmeyer HU, Hørder M, Rej R. Approved recommendation on IFCC methods for the measurement of catalytic concentration of enzymes. Part 2. IFCC method for aspartate aminotransferase. J Clin Chem Clin Biochem. 1986; b 24: 497-510
    PubMedGoogle Scholar
  • 21 Doumas BT, Watson WA, Biggs HG. Albumin standards and the measurement of serum albumin with bromcresol green. Clin Chim Acta. 1971; 31: 87-96
    CrossrefPubMedGoogle Scholar
  • 22 Wahlefeld AW, Herz G, Bernt E. Modification of the Malloy-Evelyn-method for a simple, reliable determination of total bilirubin in serum. Scand J Clin Lab Invest. 1972; 29 (Suppl. 126) Abstr. 11.12
    PubMedGoogle Scholar
  • 23 Menon KV, Kamath PS. Managing the complications of cirrhosis. Mayo Clin Proc. 2000; 75: 501-509
    CrossrefPubMedGoogle Scholar
  • 24 Bosch J, Garcia-Pagan JC. Complications of Cirrhosis. I. Portal hypertension. J Hepatol. 2000; 32 (1 Suppl) 141-156
    CrossrefPubMedGoogle Scholar
  • 25 Zhang J, Pegoli jr W, Clemens M. Endothelin-1 induces direct constriction of hepatic sinusoids. Am J Physiol. 1994; 266: G624-G632
    PubMedGoogle Scholar
  • 26 Rockey DC, Weisinger RA. Endothelin induced contractility of stellate cells from normal and cirrhotic rat liver: implications for regulation of portal pressure and resistance. Hepatology. 1996; 24: 233-240
    CrossrefPubMedGoogle Scholar
  • 27 Rockey D, Chung J. Reduced nitric oxide production by endothelial cells in cirrhotic rat liver: Endothelial dysfunction in portal hypertension. Gastroenterology. 1996; 114: 344-351
    PubMedGoogle Scholar
  • 28 Bathal PS, Grossmann HJ. Reduction of the increased portal vascular resistance of the isolated perfused cirrhotic rat liver by vasodilators. J Hepatol. 1985; 1: 325-329
    CrossrefPubMedGoogle Scholar
  • 29 Grose RD, Plevris JN, Redhead DN, Bouchier IAD, Hayes P. The acute and chronic effects of isosorbide-5-mononitrate on portal hemodynamics in cirrhosis. J Hepatol. 1994; 20: 542-547
    CrossrefPubMedGoogle Scholar
  • 30 Mela M, Mancuso A, Burroughs A. Drug treatment for portal hypertension. Ann Hepatol. 2002; 3: 102-120
    PubMedGoogle Scholar
  • 31 Wiest R, Groszmann RJ. The paradox of nitric oxide in cirrhosis and portal hypertension: too much, not enough. Hepatology. 2002; 35: 478-491
    CrossrefPubMedGoogle Scholar
  • 32 Gentilini P, La Villa G, Laffi G, Pinzani M. Hemodynamic derangement and cardiac dysfunction in cirrhosis. Ann Ital Med Int. 2004; 19: 90-97
    PubMedGoogle Scholar
  • 33 Reeves HL, Friedman SL. Activation of hepatic stellate cells-a-key issue in liver fibrosis. Front Biosc. 2002; 7: d808-d826
    CrossrefPubMedGoogle Scholar
  • 34 Murphy F, Arthur M, Iredale J. Developing strategies for liver fibrosis treatment. Expert Opin Invest Drugs. 2002; 11: 1575-1585
    CrossrefPubMedGoogle Scholar
  • 35 Gupta TK, Toruner M, Chung MK, Groszmann RJ. Endothelial dysfunction and decreased production of nitric oxide in the intrahepatic microcirculation of cirrhotic rats. Hepatology. 1998; 28: 926-931
    CrossrefPubMedGoogle Scholar
  • 36 Hobbs AJ. Soluble guanylate cyclase: an old therapeutic target revisited. Br J Pharmacol. 2002; 136: 637-640
    CrossrefPubMedGoogle Scholar
  • 37 Tulis DA. Salutary properties of YC-1 in the cardiovascular and hematological systems. Curr Med Chem. 2004; 2: 343-359
    PubMedGoogle Scholar
  • 38 Minin EA, Buchalov IB, Wellner M, Palmes D, Spiegel H-U, Neumann J et al. L-Arginine-NO-cGMP signaling following acute liver injury in the rat. Exp Tox Path. 2005; 57: 161-171
    PubMedGoogle Scholar
  • 39 Fiorucci S, Antonelli E, Morelli O, Mencarelli A, Casini A, Mello T et al. NCX-1000, a NO-releasing derivative of urodesoxcholic acid, selectively delivers NO to the liver and protects against development of portal hypertension. Proc Natl Acad Sci. 2002; 98: 8897-8902
    PubMedGoogle Scholar
  • 40 Fiorucci S, Antonelli E, Tocchetti P, Morelli A. Treatment of portal hypertension with NCX-1000, a liver-specific NO donor. A review of its current status. Cardiovasc Drug Rev. 2004; 22: 135-146
    PubMedGoogle Scholar
  • 41 Fiorucci S, Antonelli E, Brancaleone V, Sanpaolo L, Orlandi S, Distrutti E. NCX-1000, a nitric oxide-releasing derivative of orsodeoxycholic acid, ameliorates portal hypertension and lowers norepinephrine-induced intrahepatic resistance in the isolated and perfused rat liver. J Hepatol. 2003; 39: 932-939
    CrossrefPubMedGoogle Scholar
  • 42 Loureiro-Silva M, Cadelina G, Iwakiri Y, Groszmann R. A liver-specific nitric oxide donor improves the intra-hepatic vascular response to both portal blood flow increase and methoxamine in cirrhotic rats. J Hepatol. 2003; 39: 940-946
    CrossrefPubMedGoogle Scholar
  • 43 Moal F, Veal N, Vuillemin E, Barrière E, Wang J, Fizanne L. Hemodynamic and antifibrotic effects of a selective liver nitric oxide donor V-PYRRO/NO in bile duct ligated rats. World J Gastroenterol. 2006; 12: 6639-6645
    PubMedGoogle Scholar
  • 44 Bosch J. Decreasing hepatic vascular tone by liver-specific NO donors: wishful thinking or a promising reality?. J Hepatol. 2003; 39: 1072-1075
    CrossrefPubMedGoogle Scholar
  • 45 Garcia-Pagan JC, Navasa M, Bosch J, Bru C, Pizcueta P, Rodes J. Enhancement of portal pressure reduction by the association of isosorbide-5- mononitrate to propranolol admionistration in patients with cirrhosis. Hepatology. 1990; 11: 230-238
    CrossrefPubMedGoogle Scholar
  • 46 Bellis l, Berzigotti A, Abraldes JG, Moitinho E, Garcia-Pagan JC, Bosch J. Low doses of isosorbide mononitrate attenuate the postprandial increase in portal pressure in patients with cirrhosis. Hepatology. 2003; 37: 378-384
    CrossrefPubMedGoogle Scholar
  • 47 Garcia-Pagan JC, Morillas R, Banares R, Albillos A, Villanueva C, Vila C. Spanish Variceal Bleeding Study Group: Propranolol plus placebo versus propranolol plus isosorbide-5 mononitrate in the prevention of a first variceal blee: A double-blind RCT. Hepatology. 2003; 37: 1260-1266
    CrossrefPubMedGoogle Scholar
  • 48 Merkel C, Marin R, Sacerdoti D, Donada C, Cavallarin G, Torboli P. Long-term results of a clinical trial of nadolol with or without isosorbide mononitrate for primary prophylaxis of variceal bleeding in cirrhosis. Hepatology. 2000; 31: 324-329
    CrossrefPubMedGoogle Scholar
  • 49 Li J, Billiar TR. Nitric oxide. IV. Determinants of nitric oxide protection and toxicity in liver. Am J Physiol. 1999; 276: G1069-G1073
    PubMedGoogle Scholar
  • 50 Hon WM, Lee KH, Khoo HE. Nitric oxide in liver diseases:friendfoe or just passerby?. Ann NY Acad Sci. 2002; 962: 275-295
    CrossrefPubMedGoogle Scholar
  • 51 Chen T, Zamora R, Zuckerbraun B, Billiar TR. Role of nitric oxide in liver injury. Curr Mol Med. 2003; 3: 519-526
    CrossrefPubMedGoogle Scholar
  • 52 Dudenhoefer AA, Loureiro-Silva MR, Cadelina GW, Gupta T, Groszman RJ. Bioactivation of nitroglycerin and vasomotor response to nitric oxide are impaired in cirrhotic rat livers. Hepatology. 2002; 36: 381-385
    CrossrefPubMedGoogle Scholar