USOO7671085B2 (12) United States Patent (10) Patent No.: US 7,671,085 B2 DOWnes et al. (45) Date of Patent: Mar. 2, 2010 (54) NON-STEROIDAL FARNESOIDX RECEPTOR 4,265,874 A 5/1981 Bonsen et al. MODULATORS AND METHODS FOR THE 5,151,442 A 9, 1992 Garcia et al. USE THEREOF FOREIGN PATENT DOCUMENTS (75) Inventors: Michael R. Downes, San Diego, CA WO WOOO/37077 6, 2000 (US); Ronald M. Evans, La Jolla, CA WO WOOOf 76.523 12/2000 (US) WO WO 2004/046.162 6, 2004 OTHER PUBLICATIONS (73) Assignee: The Salk Institute for Biological Fi1Orucci iS. S, RizzoRIZZO (J,G. DoniniJonini A,A. DistruttiJS 1 E.E, and SanlucciSantucci L.L, “Targetilargeting Studies, La Jolla, CA (US) farnesoid X receptor for liver and metabolic disorders.” Trends in c - r Molecular Medicine, Jul. 2007. 13(7) 298–309.* (*) Notice: Subj ect to any disclaimer, the term of this Scotti E, Gilardi F. Godio C, Gers E, Krneta J. Mitro N. De Fabiani E. patent is extended or adjusted under 35 Caruso D, and Crestani M. "Bile acids and their signaling pathways: U.S.C. 154(b) by 907 days. eclectic regulators of diverse cellular functions.” Cellular and Molecular Life Sciences, Oct. 2007, 64(19-20), 2477-2491.* (21) Appl. No.: 10/535,043 Laffitte et al., “Identification of the DNA binding specificity and potential target genes for the farnesoid X-activated receptor.” Journal (22) PCT Filed: Nov. 14, 2003 of Biological Chemistry, 275: 10638-10647, 2000. Nicolaou et al., “Natural product-like combinatorial libraries based (86). PCT No.: PCT/USO3/36137 on privileged structures. 1. General principles and Solid-phase syn thesis of benzopyrians.” Journal of the American Chem. Soc., 122: S371 (c)(1), 9939-9953, 2000. (2), (4) Date: Dec. 9, 2005 Nicolaou et al., “Natural product-like combinatorial libraries based s 9 on privileged structures. 2. Construction of a 10,000-membered benzopyran library by directed split-and-pool chemistry using (87) PCT Pub. No.: WO2004/046068 nanoKans and optical encoding.” Journal of the American Chem. PCT Pub. Date: Jun. 3, 2004 Soc., 122:9954-9967, 2000. * cited by examiner (65) Prior Publication Data Primary Examiner San-ming Hui US 2006/O128764 A1 Jun. 15, 2006 Assistant Examiner Paul Zarek O O (74) Attorney, Agent, or Firm—Foley & Lardner LLP: Related U.S. Application Data Stephen E. Reiter (63) Continuation-in-part of application No. 10/658,115, filed on Sep. 8, 2003, now abandoned. (57) ABSTRACT (60) Provisional application No. 60/426,664, filed on Nov. The efficient regulation of cholesterol synthesis, metabolism, 15, 2002. acquisition, and transport is an essential component of lipid homeostasis. The farnesoid X receptor (FXR) is a transcrip 51) Int. Cl. tional sensor for bile acids, the primprimary product of cholesterol AOIN 43/16 2006.O1 metabolism. Accordingly,gly the developmentp of potent,p selec AOIN 37/2 (2006.01) tive, Small molecule agonists, partial agonists, and antago AOIN 37/44 (2006.01) nists of FXR would be an important step in further deconvo A6 IK3I/35 (2006.01) luting FXR physiology. In accordance with the present A6 IK 3L/24 (2006.01) invention, the identification of novel potent FXRactivators is (52) U.S. Cl. ........................ 514/456; 514/539; 514/620 described. Two derivatives of invention compounds, bearing (58) Field of Classification Search ....................... None stilbene orbiaryl moieties, contain members that are the most See application file for complete search history. potent FXR agonists reported to date in cell-based assays. These compounds are useful as chemical tools to further (56) References Cited define the physiological role of FXR as well as therapeutic U.S. PATENT DOCUMENTS leads for the treatment of diseases linked to cholesterol, bile acids and their metabolism and homeostasis. 4,160,452 A 7, 1979 Theeuwes 4,256,108 A 3, 1981 Theeuwes 18 Claims, 1 Drawing Sheet U.S. Patent Mar. 2, 2010 US 7,671,085 B2 NHR response element Cell based Assay KRRXR on a FKRRE 35000 s otSC Grah uth lul M Conc of CDCA FXR efficacy on a 384 well plate. Figure 1 US 7,671,085 B2 1. 2 NON-STEROIDAL FARNESOIDX RECEPTOR binds to LRH-1 (liver receptor homolog), which is required in MODULATORS AND METHODS FOR THE CYP7A activation. Additionally, both LXR and FXR are USE THEREOF implicated in the regulation of several other gene products involved in cholesterol absorption, metabolism and transport. FIELD OF THE INVENTION Thus, the identification of potent, selective, small molecule FXRagonists, partial agonists and antagonists would be pow The present invention relates to new chemical entities. In a erful tools and would have many potential applications. For particular aspect, the present invention relates to non-steroi example, Such compounds would facilitate the in vivo analy dal modulators of farnesoid X receptors (FXR). In another sis of FXR physiology in vivo. In addition, Such compounds, aspect, the present invention relates to methods for modulat 10 in conjunction with DNA arraying technology, might allow ing FXR-mediated processes employing the novel com for the discovery of new gene products under the control of pounds described herein. FXR. Further, FXR modulators might find potential utility in the treatment of cholestasis and other disease states associ BACKGROUND OF THE INVENTION ated with aberrant levels, flow and release of bile acids. More 15 over, in the absence of a crystal structure of FXR, a thorough The following discussion of the background of the inven structure-activity relationship (SAR) study of ligands that tion is merely provided to aid the reader in understanding the modulate the activity of FXR would allow for the delineation invention and is not admitted to describe or constitute prior art of the structural requirements for ligand binding and might to the present invention. aid in the design of future ligands and potential therapeutics. The efficient regulation of cholesterol biosynthesis, metabolism, acquisition and transport is an essential function SUMMARY OF THE INVENTION of mammalian cells. High levels of cholesterol are associated with atherosclerosis, a leading cause of death in the western In accordance with the present invention, the identification world and a major risk factor correlated with the occurrence of novel potent FXR activators is described. Initial screening of coronary heart disease and stroke. Until recently, recom 25 of a 10,000-membered, diversity-orientated library of ben mendations for the treatment of hypercholestemia were Zopyran containing Small molecules for FXR activation uti focused on the use of statins, which inhibit the de novo bio lizing a cell-based reporter assay led to the identification of synthesis of cholesterol, and the use of bile acid sequestering several lead compounds owning low micromolar activity agents. While statin-based agents are still in widespread use (ECss=5-10 uM. These compounds were systematically as cholesterol-lowering drugs, an evolving understanding of 30 modified employing parallel solution-phase synthesis and the mechanisms controlling cholesterol homeostasis has led Solid-phase synthesis to provide numerous compounds that to new molecular targets as candidates in therapeutic inter potently activate FXR. Two derivatives of invention com vention. pounds, bearing stilbene orbiaryl moieties, contain members Cholesterol metabolism is controlled through a complex that are the most potent FXR agonists reported to date in feedback loop involving cholesterol itself and bile acids 35 cell-based assays. These compounds are useful as chemical (which are primary oxidation products), and through secre tools to further define the physiological role of FXR as well as tion in the gut, the single most critical regulators of choles therapeutic leads for the treatment of diseases linked to cho terol absorption. The nuclear receptors LXR (liver X recep lesterol, bile acids and their metabolism and homeostasis. tor) and FXR (farnesoid X receptor) are the specialized sensors of cholesterol and bile acids that control transcription 40 BRIEF DESCRIPTION OF THE FIGURE of networks encoding key metabolic enzymes. For example activation of LXR by oxysterols (i.e., mono-oxygenated cho FIG. 1 summarizes the efficacy of the functional assay for lesterol metabolites) leads to the up-regulation of CYP7A1, the identification of FXR agonists, using the known FXR the enzyme that catalyzes the rate limiting step in the conver agonist, chenodeoxycholic acid (CDCA). sion of cholesterol to bile acids. In turn, bile acids such as 45 chenodeoxycholic acid (CDCA, 1, a low affinity endogenous DETAILED DESCRIPTION OF THE INVENTION agonist for FXR, whose structure is shown below) are potent ligands for FXR, whose activation leads to down-regulation In accordance with the present invention, there are pro of CYP7A1, leading to the completion of the feedback cir vided compounds having the structure: cuit. 50 55 60 21y-OR wherein: A is a C3 up to C8 branched chain alkyl or substituted alkyl 65 group, a C3 up to C7 cycloalkyl or Substituted cycloalkyl, an In this circuit FXR induces the expression of a transcriptional optionally substituted aryl or an optionally substituted het repressor SHP (small heterodimer partner) which in turn eroaryl, US 7,671,085 B2 3 4 X is —C(O)— or —CH2—, tuted heterocyclic” refers to heterocyclic groups further bear R is methyl or ethyl, ing one or more Substituents as set forth above. R" is H, hydroxy, alkoxy, benzoyloxy, mesityloxy, or As employed herein, “aryl” refers to aromatic groups hav –OCHC(O)OCHs. ing in the range of 6 up to 14 carbon atoms and “substituted R’ is Hor R can cooperate with R to form a benzopyran, 5 aryl” refers to aryl groups further bearing one or more Sub wherein the pyran ring has the structure: stituents as set forth above. As employed herein, “aryloxy' refers to —O-aryl groups having in the range of 6 up to 14 carbon atoms and 'substi tuted aryloxy' refers to aryloxy groups further bearing one or 10 more substituents as set forth above.