Inhibition of TGF-beta signaling by an ALK5 inhibitor protects rats from dimethylnitrosamine-induced liver fibrosis.
1 Chronic liver disease is characterized by an exacerbated accumulation of matrix, causing progressive fibrosis, which may lead to cirrhosis. Transforming growth factor beta (TGF-beta), a well-known profibrotic cytokine, transduces its signal through the ALK5 ser/thr kinase receptor, and increases transcription of different genes including PAI-1 and collagens. The identification of GW6604 (2-phenyl-4-(3-pyridin-2-yl-1H-pyrazol-4-yl)pyridine), an ALK5 inhibitor, allowed us to evaluate the therapeutic potential of inhibiting TGF-beta pathway in different models of liver disease. 2 A cellular assay was used to identify GW6604 as a TGF-beta signaling pathway inhibitor. This ALK5 inhibitor was then tested in a model of liver hepatectomy in TGF-beta-overexpressing transgenic mice, in an acute model of liver disease and in a chronic model of dimethylnitrosamine (DMN)-induced liver fibrosis. 3 In vitro, GW6604 inhibited autophosphorylation of ALK5 with an IC(50) of 140 nM and in a cellular assay inhibited TGF-beta-induced transcription of PAI-1 (IC(50): 500 nM). In vivo, GW6604 (40 mg kg(-1) p.o.) increased liver regeneration in TGF-beta-overexpressing mice, which had undergone partial hepatectomy. In an acute model of liver disease, GW6604 reduced by 80% the expression of collagen IA1. In a chronic model of DMN-induced fibrosis where DMN was administered for 6 weeks and GW6604 dosed for the last 3 weeks (80 mg kg(-1) p.o., b.i.d.), mortality was prevented and DMN-induced elevations of mRNA encoding for collagen IA1, IA2, III, TIMP-1 and TGF-beta were reduced by 50-75%. Inhibition of matrix genes overexpression was accompanied by reduced matrix deposition and reduction in liver function deterioration, as assessed by bilirubin and liver enzyme levels. 4 Our results suggest that inhibition of ALK5 could be an attractive new approach to treatment of liver fibrotic diseases by both preventing matrix deposition and promoting hepatocyte regeneration.
de Gouville
Anne-Charlotte
AC
Biology Department, GlaxoSmithKline, 25 avenue du Quebec, Les Ulis 91951, France. m.degouville@gsk.com
Boullay
Valerie
V
Krysa
Gael
G
Pilot
Julia
J
Brusq
Jean-Marie
JM
Loriolle
Florence
F
Gauthier
Jean-Michel
JM
Papworth
Stephen A
SA
Laroze
Alain
A
Gellibert
Françoise
F
Huet
Stephane
S
D030201:Activin Receptors, Type I / Q000037:antagonists & inhibitors* / Q000378:metabolism; D000208:Acute Disease; D000818:Animals; D045744:Cell Line, Tumor; D002908:Chronic Disease; D004128:Dimethylnitrosamine; D006498:Hepatectomy; D006801:Humans; D008103:Liver Cirrhosis / Q000139:chemically induced / Q000473:pathology / Q000517:prevention & control*; D008115:Liver Regeneration / Q000187:drug effects; D008297:Male; D051379:Mice; D008822:Mice, Transgenic; D010766:Phosphorylation; D017395:Plasminogen Activator Inhibitor 1 / Q000096:biosynthesis / Q000235:genetics; D017346:Protein Serine-Threonine Kinases; D011720:Pyrazoles / Q000494:pharmacology*; D011725:Pyridines / Q000494:pharmacology*; D051381:Rats; D017207:Rats, Sprague-Dawley; D000077293:Receptor, Transforming Growth Factor-beta Type I; D018125:Receptors, Transforming Growth Factor beta / Q000037:antagonists & inhibitors* / Q000378:metabolism; D015398:Signal Transduction; D014158:Transcription, Genetic; D016212:Transforming Growth Factor beta / Q000037:antagonists & inhibitors* / Q000235:genetics / Q000502:physiology
C502971:2-phenyl-4-(3-pyridin-2-yl-1H-pyrazol-4-yl)pyridine; D017395:Plasminogen Activator Inhibitor 1; D011720:Pyrazoles; D011725:Pyridines; D018125:Receptors, Transforming Growth Factor beta; D016212:Transforming Growth Factor beta; D017346:Protein Serine-Threonine Kinases; D030201:Activin Receptors, Type I; D000077293:Receptor, Transforming Growth Factor-beta Type I; C000625841:TGFBR1 protein, human; C000625842:Tgfbr1 protein, mouse; C000625843:Tgfbr1 protein, rat; D004128:Dimethylnitrosamine
EMBO J. 1998 Jun 1;17(11):3091-100
9606191
Arch Pathol Lab Med. 1976 Aug;100(8):405-14
60092
Front Neuroendocrinol. 1998 Oct;19(4):287-322
9799587
Arch Pathol. 1964 Jul;78:69-72
14150734
Histochemistry. 1986;85(2):129-33
3744896
J Histochem Cytochem. 1985 Aug;33(8):737-43
2410480
Cell. 2003 Jun 13;113(6):685-700
12809600
Mol Pharmacol. 2002 Jul;62(1):65-74
12065756
Hepatol Res. 2002 Oct;24(2):174
12270747
Proc Natl Acad Sci U S A. 1995 Mar 28;92(7):2572-6
7708687
Proc Natl Acad Sci U S A. 2000 Jul 5;97(14):8015-20
10859350
Hepatology. 2000 Aug;32(2):247-55
10915731
Kidney Int. 1998 Nov;54(5):1570-80
9844133
J Clin Invest. 1996 Jan 1;97(1):232-7
8550840
J Biol Chem. 1991 Jan 15;266(2):1092-100
1985937
Science. 2002 May 31;296(5573):1646-7
12040180
Scand J Clin Lab Invest. 1999 Aug;59(5):343-7
10533846
Proc Natl Acad Sci U S A. 1991 Aug 1;88(15):6642-6
1862087
J Med Chem. 2002 Feb 28;45(5):999-1001
11855979
EMBO J. 1987 May;6(5):1281-6
3111844
J Clin Pathol. 1996 Dec;49(12):998-1004
9038738
J Hepatol. 1999 Oct;31(4):703-11
10551395
Thorax. 1999 Sep;54(9):805-12
10456973
J Hepatol. 2002 Feb;36(2):200-9
11830331
N Engl J Med. 1994 Nov 10;331(19):1286-92
7935686
Hepatology. 2003 Nov;38(5):1107-15
14578849
Cell. 1999 Feb 5;96(3):425-36
10025408
BMC Gastroenterol. 2003 Oct 18;3:29
14565855
Mol Pharmacol. 2002 Jul;62(1):58-64
12065755
Am J Pathol. 1999 Apr;154(4):1115-24
10233850
Toxicology. 2001 Jan 2;156(2-3):129-38
11164615
Mol Biol Cell. 2000 Feb;11(2):555-65
10679014
Matrix. 1992 Feb;12(1):36-43
1560788
J Hepatol. 1994 Mar;20(3):388-93
8014451
J Gastroenterol Hepatol. 1999 Jul;14(7):618-33
10440206
N Engl J Med. 2000 May 4;342(18):1350-8
10793168
Proc Natl Acad Sci U S A. 1992 Jun 15;89(12):5408-12
1608949
Lab Invest. 1996 Jun;74(6):991-1003
8667617
Mol Reprod Dev. 1992 Jun;32(2):111-20
1637549
J Med Chem. 2003 Sep 11;46(19):3953-6
12954047
Proc Natl Acad Sci U S A. 1996 Jun 11;93(12):5877-82
8650186
Cell. 2000 Oct 13;103(2):295-309
11057902
Clin Biochem. 2000 Oct;33(7):563-70
11124342
Cell. 1993 Nov 19;75(4):681-92
8242743
J Hepatol. 1998 May;28(5):803-11
9625315
Scand J Gastroenterol. 1996 Dec;31(12):1137-43
8976003
J Viral Hepat. 2001 Nov;8(6):430-7
11703574
Am J Respir Crit Care Med. 2001 Mar;163(3 Pt 1):770-7
11254537
J Biol Chem. 2002 Mar 29;277(13):11069-76
11796725