Synthetic Farnesoid X Receptor Agonists Induce High-Density Lipoprotein-Mediated Transhepatic Cholesterol Efflux in Mice and Monkeys and Prevent Atherosclerosis in Cholesteryl Ester Transfer Protein Transgenic Low-Density Lipoprotein Receptor (−/−) Mice

Farnesoid X receptor (FXR), a bile acid-activated nuclear hormone receptor, plays an important role in the regulation of cholesterol and more specifically high-density lipoprotein (HDL) homeostasis. Activation of FXR is reported to lead to both pro- and anti-atherosclerotic effects. In the present study we analyzed the impact of different FXR agonists on cholesterol homeostasis, plasma lipoprotein profiles, and transhepatic cholesterol efflux in C57BL/6J mice and cynomolgus monkeys and atherosclerosis development in cholesteryl ester transfer protein transgenic (CETPtg) low-density lipoprotein receptor (LDLR) (−/−) mice. In C57BL/6J mice on a high-fat diet the synthetic FXR agonists isopropyl 3-(3,4-difluorobenzoyl)-1,1-dimethyl-1,2,3,6-tetrahydroazepino[4,5-b]indole-5-carboxylate (FXR-450) and 4-[2-[2-chloro-4-[[5-cyclopropyl-3-(2,6-dichlorophenyl)-4-isoxazolyl]methoxy]phenyl]cyclopropyl]benzoic acid (PX20606) demonstrated potent plasma cholesterol-lowering activity that affected all lipoprotein species, whereas 3-[2-[2-chloro-4-[[3-(2,6-dichlorophenyl)-5-(1-methylethyl)-4-isoxazolyl]methoxy]phenyl]ethenyl]benzoic acid (GW4064) and 6-ethyl chenodeoxycholic acid (6-ECDCA) showed only limited effects. In FXR wild-type mice, but not FXR(−/−) mice, the more efficacious FXR agonists increased fecal cholesterol excretion and reduced intestinal cholesterol (re)uptake. In CETPtg-LDLR(−/−) mice PX20606 potently lowered total cholesterol and, despite the observed HDL cholesterol (HDLc) reduction, caused a highly significant decrease in atherosclerotic plaque size. In normolipidemic cynomolgus monkeys PX20606 and 6-ECDCA both reduced total cholesterol, and PX20606 specifically lowered HDL2c but not HDL3c or apolipoprotein A1. That pharmacological FXR activation specifically affects this cholesterol-rich HDL2 subclass is a new and highly interesting finding and sheds new light on FXR-dependent HDLc lowering, which has been perceived as a major limitation for the clinical development of FXR agonists.

[1]  Barbara Gross,et al.  Bile acid receptors as targets for the treatment of dyslipidemia and cardiovascular disease , 2012, Journal of Lipid Research.

[2]  Ann M. Thomas,et al.  Farnesoid X Receptor Induces Murine Scavenger Receptor Class B Type I via Intron Binding , 2012, PloS one.

[3]  P. Tontonoz,et al.  Transcriptional integration of metabolism by the nuclear sterol-activated receptors LXR and FXR , 2012, Nature Reviews Molecular Cell Biology.

[4]  P. Barter,et al.  Predictive value of different HDL particles for the protection against or risk of coronary heart disease. , 2012, Biochimica et biophysica acta.

[5]  J. Chaparro-Riggers,et al.  Proprotein Convertase Substilisin/Kexin Type 9 Antagonism Reduces Low-Density Lipoprotein Cholesterol in Statin-Treated Hypercholesterolemic Nonhuman Primates , 2012, Journal of Pharmacology and Experimental Therapeutics.

[6]  A. Kontush,et al.  Biological activities of HDL subpopulations and their relevance to cardiovascular disease. , 2011, Trends in molecular medicine.

[7]  M. Yeh,et al.  Hepatic free cholesterol accumulates in obese, diabetic mice and causes nonalcoholic steatohepatitis. , 2011, Gastroenterology.

[8]  Hans Richter,et al.  Studies in mice, hamsters, and rats demonstrate that repression of hepatic apoA-I expression by taurocholic acid in mice is not mediated by the farnesoid-X-receptor , 2011, Journal of Lipid Research.

[9]  D. Rader,et al.  Novel HDL-directed pharmacotherapeutic strategies , 2011, Nature Reviews Cardiology.

[10]  S. Kliewer,et al.  FGF19 as a Postprandial, Insulin-Independent Activator of Hepatic Protein and Glycogen Synthesis , 2011, Science.

[11]  Robert L Wilensky,et al.  Cholesterol efflux capacity, high-density lipoprotein function, and atherosclerosis. , 2011, The New England journal of medicine.

[12]  W. Gong,et al.  Upregulation of scavenger receptor class B type I expression by activation of FXR in hepatocyte. , 2010, Atherosclerosis.

[13]  U. Deuschle,et al.  Synthesis and pharmacological validation of a novel series of non-steroidal FXR agonists. , 2010, Bioorganic & medicinal chemistry letters.

[14]  F. Kuipers,et al.  A Role of the Bile Salt Receptor FXR in Atherosclerosis , 2010, Arteriosclerosis, thrombosis, and vascular biology.

[15]  M. L. Crawley,et al.  Farnesoid X receptor modulators: a patent review , 2010, Expert Opinion on Therapeutic Patents.

[16]  M. Miyazaki,et al.  Farnesoid X Receptor Activation Prevents the Development of Vascular Calcification in ApoE−/− Mice With Chronic Kidney Disease , 2010, Circulation research.

[17]  M. Trauner,et al.  Bile Acids as Regulators of Hepatic Lipid and Glucose Metabolism , 2010, Digestive Diseases.

[18]  T. Willson,et al.  Identification of Novel Pathways That Control Farnesoid X Receptor-mediated Hypocholesterolemia* , 2009, The Journal of Biological Chemistry.

[19]  M. Evans,et al.  Pyrrole[2,3-d]azepino compounds as agonists of the farnesoid X receptor (FXR). , 2009, Bioorganic & medicinal chemistry letters.

[20]  M. Evans,et al.  Activation of farnesoid X receptor prevents atherosclerotic lesion formation in LDLR−/− and apoE−/− mice Published, JLR Papers in Press, January 27, 2009. , 2009, Journal of Lipid Research.

[21]  U. Tietge,et al.  Scavenger Receptor BI-mediated Selective Uptake Is Required for the Remodeling of High Density Lipoprotein by Endothelial Lipase* , 2009, Journal of Biological Chemistry.

[22]  C. Huard,et al.  A synthetic farnesoid X receptor (FXR) agonist promotes cholesterol lowering in models of dyslipidemia. , 2009, American journal of physiology. Gastrointestinal and liver physiology.

[23]  E. Distrutti,et al.  Antiatherosclerotic effect of farnesoid X receptor. , 2009, American journal of physiology. Heart and circulatory physiology.

[24]  P. Edwards,et al.  FXR signaling in metabolic disease , 2008, FEBS letters.

[25]  T. Warner,et al.  Farnesoid X Receptor Ligands Inhibit Vascular Smooth Muscle Cell Inflammation and Migration , 2007, Arteriosclerosis, thrombosis, and vascular biology.

[26]  J. Auwerx,et al.  In vivo imaging of farnesoid X receptor activity reveals the ileum as the primary bile acid signaling tissue. , 2007, Molecular endocrinology.

[27]  B. Brewer,et al.  Effects of FXR in foam-cell formation and atherosclerosis development. , 2006, Biochimica et biophysica acta.

[28]  P. Edwards,et al.  FXR Deficiency Causes Reduced Atherosclerosis in Ldlr−/− Mice , 2006, Arteriosclerosis, thrombosis, and vascular biology.

[29]  Folkert Kuipers,et al.  The Farnesoid X Receptor Modulates Adiposity and Peripheral Insulin Sensitivity in Mice* , 2006, Journal of Biological Chemistry.

[30]  Ke Ma,et al.  Farnesoid X receptor is essential for normal glucose homeostasis. , 2006, The Journal of clinical investigation.

[31]  T. Willson,et al.  Activation of the nuclear receptor FXR improves hyperglycemia and hyperlipidemia in diabetic mice. , 2006, Proceedings of the National Academy of Sciences of the United States of America.

[32]  M. Reilly,et al.  Endothelial Lipase Concentrations Are Increased in Metabolic Syndrome and Associated with Coronary Atherosclerosis , 2005, PLoS medicine.

[33]  M. Reilly,et al.  Hepatic expression of scavenger receptor class B type I (SR-BI) is a positive regulator of macrophage reverse cholesterol transport in vivo. , 2005, The Journal of clinical investigation.

[34]  S. Kliewer,et al.  Fibroblast growth factor 15 functions as an enterohepatic signal to regulate bile acid homeostasis. , 2005, Cell metabolism.

[35]  V. Rodríguez-Sureda,et al.  A procedure for measuring triacylglyceride and cholesterol content using a small amount of tissue. , 2005, Analytical biochemistry.

[36]  Jonathan C. Cohen,et al.  Expression of ABCG5 and ABCG8 Is Required for Regulation of Biliary Cholesterol Secretion* , 2005, Journal of Biological Chemistry.

[37]  R. Savkur,et al.  Regulation of carbohydrate metabolism by the farnesoid X receptor. , 2005, Endocrinology.

[38]  N. Webb,et al.  Remodeling of HDL remnants generated by scavenger receptor class B type I Published, JLR Papers in Press, June 21, 2004. DOI 10.1194/jlr.M400026-JLR200 , 2004, Journal of Lipid Research.

[39]  T. Warner,et al.  Expression and activation of the farnesoid X receptor in the vasculature. , 2004, Proceedings of the National Academy of Sciences of the United States of America.

[40]  R. Blevins,et al.  Farnesoid X Receptor Activates Transcription of the Phospholipid Pump MDR3* , 2003, Journal of Biological Chemistry.

[41]  Grace Guo,et al.  The Farnesoid X-receptor Is an Essential Regulator of Cholesterol Homeostasis* , 2003, The Journal of Biological Chemistry.

[42]  M. Kindy,et al.  The fate of HDL particles in vivo after SR-BI-mediated selective lipid uptake Published, JLR Papers in Press, August 16, 2002. DOI 10.1194/jlr.M200173-JLR200 , 2002, Journal of Lipid Research.

[43]  T. Willson,et al.  6alpha-ethyl-chenodeoxycholic acid (6-ECDCA), a potent and selective FXR agonist endowed with anticholestatic activity. , 2002, Journal of medicinal chemistry.

[44]  J. Dallongeville,et al.  Bile acid-activated nuclear receptor FXR suppresses apolipoprotein A-I transcription via a negative FXR response element. , 2002, The Journal of clinical investigation.

[45]  T. Willson,et al.  Chemical genomics: Functional analysis of orphan nuclear receptors in the regulation of bile acid metabolism , 2001, Medicinal research reviews.

[46]  M. Makishima,et al.  Human Bile Salt Export Pump Promoter Is Transactivated by the Farnesoid X Receptor/Bile Acid Receptor* , 2001, The Journal of Biological Chemistry.

[47]  D. Moore,et al.  The Farnesoid X-activated Receptor Mediates Bile Acid Activation of Phospholipid Transfer Protein Gene Expression* , 2000, The Journal of Biological Chemistry.

[48]  Masahiro Tohkin,et al.  Targeted Disruption of the Nuclear Receptor FXR/BAR Impairs Bile Acid and Lipid Homeostasis , 2000, Cell.

[49]  L. Moore,et al.  A regulatory cascade of the nuclear receptors FXR, SHP-1, and LRH-1 represses bile acid biosynthesis. , 2000, Molecular cell.

[50]  T. A. Kerr,et al.  Molecular basis for feedback regulation of bile acid synthesis by nuclear receptors. , 2000, Molecular cell.

[51]  L. Moore,et al.  Identification of a chemical tool for the orphan nuclear receptor FXR. , 2000, Journal of medicinal chemistry.

[52]  D. Rader,et al.  Gene transfer and hepatic overexpression of the HDL receptor SR-BI reduces atherosclerosis in the cholesterol-fed LDL receptor-deficient mouse. , 2000, Arteriosclerosis, thrombosis, and vascular biology.

[53]  J. Lehmann,et al.  Bile acids: natural ligands for an orphan nuclear receptor. , 1999, Science.

[54]  M. Makishima,et al.  Identification of a nuclear receptor for bile acids. , 1999, Science.

[55]  E. Rubin,et al.  Lower Plasma Levels and Accelerated Clearance of High Density Lipoprotein (HDL) and Non-HDL Cholesterol in Scavenger Receptor Class B Type I Transgenic Mice* , 1999, The Journal of Biological Chemistry.

[56]  A. Tall,et al.  Decreased Atherosclerosis in Heterozygous Low Density Lipoprotein Receptor-deficient Mice Expressing the Scavenger Receptor BI Transgene* , 1999, The Journal of Biological Chemistry.

[57]  E. Edelman,et al.  Overexpression of the HDL receptor SR-BI alters plasma HDL and bile cholesterol levels , 1997, Nature.

[58]  A. Tall,et al.  Dietary cholesterol increases transcription of the human cholesteryl ester transfer protein gene in transgenic mice. Dependence on natural flanking sequences. , 1992, The Journal of clinical investigation.

[59]  A. Tall,et al.  Reduced high density lipoprotein cholesterol in human cholesteryl ester transfer protein transgenic mice. , 1991, The Journal of biological chemistry.

[60]  S. Manabe,et al.  Preliminary dose finding study for subacute toxicological study of pravastatin sodium in monkeys. , 1989, The Journal of toxicological sciences.

[61]  S. Goldstein,et al.  A density gradient ultracentrifugal procedure for the isolation of the major lipoprotein classes from human serum. , 1981, Journal of lipid research.

[62]  N S Radin,et al.  Lipid extraction of tissues with a low-toxicity solvent. , 1978, Analytical biochemistry.

[63]  S. Grundy,et al.  QUANTITATIVE ISOLATION AND GAS--LIQUID CHROMATOGRAPHIC ANALYSIS OF TOTAL FECAL BILE ACIDS. , 1965, Journal of lipid research.

[64]  S. Grundy,et al.  QUANTITATIVE ISOLATION AND GAS--LIQUID CHROMATOGRAPHIC ANALYSIS OF TOTAL DIETARY AND FECAL NEUTRAL STEROIDS. , 1965, Journal of lipid research.

[65]  B. Staels,et al.  Role of bile acids and bile acid receptors in metabolic regulation. , 2009, Physiological reviews.

[66]  F. Kuipers,et al.  Increased fecal neutral sterol loss upon liver X receptor activation is independent of biliary sterol secretion in mice. , 2005, Gastroenterology.

[67]  Review Article Non-Alcoholic Fatty Liver Disease: The Bile Acid-Activated Farnesoid X Receptor as an Emerging Treatment Target , 2022 .