title: A Genomic DNA Reporter Screen Identifies Squalene Synthase Inhibitors That Act Cooperatively with Statins to Upregulate the Low-Density Lipoprotein Receptor.
abstract: Hypercholesterolemia remains one of the leading risk factors for the development of cardiovascular disease. Many large double-blind studies have demonstrated that lowering low-density lipoprotein (LDL) cholesterol using a statin can reduce the risk of having a cardiovascular event by approximately 30%. However, despite the success of statins, some patient populations are unable to lower their LDL cholesterol to meet the targeted lipid levels, due to compliance or potency issues. This is especially true for patients with heterozygous familial hypercholesterolemia who may require additional upregulation of the low-density lipoprotein receptor (LDLR) to reduce LDL cholesterol levels below those achievable with maximal dosing of statins. Here we identify a series of small molecules from a genomic DNA reporter screen that upregulate the LDLR in mouse and human liver cell lines at nanomolar potencies (EC50= 39 nM). Structure-activity relationship studies carried out on the lead compound, OX03771 [(E)-N,N-dimethyl-3-(4-styrylphenoxy)propan-1-amine], led to the identification of compound OX03050 [(E)-3-(4-styrylphenoxy)propan-1-ol], which had similar potency (EC50= 26 nM) but a much-improved pharmacokinetic profile and showed in vivo efficacy. Compounds OX03050 and OX03771 were found to inhibit squalene synthase, the first committed step in cholesterol biosynthesis. These squalene synthase inhibitors were shown to act cooperatively with statins to increase LDLR expression in vitro. Overall, we demonstrated here a novel series of small molecules with the potential to be further developed to treat patients either alone or in combination with statins.
authors.affiliation: Departments of Physiology, Anatomy, and Genetics (A.G.K., L.C.S.T., M.C., S.A., O.H., J.H.-M., R.W.-M.) and Pharmacology (A.J.R.), University of Oxford, Oxford, United Kingdom; Division of Cardiovascular Medicine, British Heart Foundation Centre of Research Excellence, University of Oxford, John Radcliffe Hospital, Oxford, United Kingdom (A.B.H., G.D., K.M.C.); and Chemistry Research Laboratory, Department of Chemistry, University of Oxford, Oxford, United Kingdom (J.M., C.J.R.B., G.M.W., A.J.R.).
authors.forename: Alastair G
authors.initials: AG
authors.lastname: Kerr
authors.affiliation: Departments of Physiology, Anatomy, and Genetics (A.G.K., L.C.S.T., M.C., S.A., O.H., J.H.-M., R.W.-M.) and Pharmacology (A.J.R.), University of Oxford, Oxford, United Kingdom; Division of Cardiovascular Medicine, British Heart Foundation Centre of Research Excellence, University of Oxford, John Radcliffe Hospital, Oxford, United Kingdom (A.B.H., G.D., K.M.C.); and Chemistry Research Laboratory, Department of Chemistry, University of Oxford, Oxford, United Kingdom (J.M., C.J.R.B., G.M.W., A.J.R.).
authors.forename: Lawrence C S
authors.initials: LCS
authors.lastname: Tam
authors.affiliation: Departments of Physiology, Anatomy, and Genetics (A.G.K., L.C.S.T., M.C., S.A., O.H., J.H.-M., R.W.-M.) and Pharmacology (A.J.R.), University of Oxford, Oxford, United Kingdom; Division of Cardiovascular Medicine, British Heart Foundation Centre of Research Excellence, University of Oxford, John Radcliffe Hospital, Oxford, United Kingdom (A.B.H., G.D., K.M.C.); and Chemistry Research Laboratory, Department of Chemistry, University of Oxford, Oxford, United Kingdom (J.M., C.J.R.B., G.M.W., A.J.R.).
authors.forename: Ashley B
authors.initials: AB
authors.lastname: Hale
authors.affiliation: Departments of Physiology, Anatomy, and Genetics (A.G.K., L.C.S.T., M.C., S.A., O.H., J.H.-M., R.W.-M.) and Pharmacology (A.J.R.), University of Oxford, Oxford, United Kingdom; Division of Cardiovascular Medicine, British Heart Foundation Centre of Research Excellence, University of Oxford, John Radcliffe Hospital, Oxford, United Kingdom (A.B.H., G.D., K.M.C.); and Chemistry Research Laboratory, Department of Chemistry, University of Oxford, Oxford, United Kingdom (J.M., C.J.R.B., G.M.W., A.J.R.).
authors.forename: Milena
authors.initials: M
authors.lastname: Cioroch
authors.affiliation: Departments of Physiology, Anatomy, and Genetics (A.G.K., L.C.S.T., M.C., S.A., O.H., J.H.-M., R.W.-M.) and Pharmacology (A.J.R.), University of Oxford, Oxford, United Kingdom; Division of Cardiovascular Medicine, British Heart Foundation Centre of Research Excellence, University of Oxford, John Radcliffe Hospital, Oxford, United Kingdom (A.B.H., G.D., K.M.C.); and Chemistry Research Laboratory, Department of Chemistry, University of Oxford, Oxford, United Kingdom (J.M., C.J.R.B., G.M.W., A.J.R.).
authors.forename: Gillian
authors.initials: G
authors.lastname: Douglas
authors.affiliation: Departments of Physiology, Anatomy, and Genetics (A.G.K., L.C.S.T., M.C., S.A., O.H., J.H.-M., R.W.-M.) and Pharmacology (A.J.R.), University of Oxford, Oxford, United Kingdom; Division of Cardiovascular Medicine, British Heart Foundation Centre of Research Excellence, University of Oxford, John Radcliffe Hospital, Oxford, United Kingdom (A.B.H., G.D., K.M.C.); and Chemistry Research Laboratory, Department of Chemistry, University of Oxford, Oxford, United Kingdom (J.M., C.J.R.B., G.M.W., A.J.R.).
authors.forename: Sarina
authors.initials: S
authors.lastname: Agkatsev
authors.affiliation: Departments of Physiology, Anatomy, and Genetics (A.G.K., L.C.S.T., M.C., S.A., O.H., J.H.-M., R.W.-M.) and Pharmacology (A.J.R.), University of Oxford, Oxford, United Kingdom; Division of Cardiovascular Medicine, British Heart Foundation Centre of Research Excellence, University of Oxford, John Radcliffe Hospital, Oxford, United Kingdom (A.B.H., G.D., K.M.C.); and Chemistry Research Laboratory, Department of Chemistry, University of Oxford, Oxford, United Kingdom (J.M., C.J.R.B., G.M.W., A.J.R.).
authors.forename: Olivia
authors.initials: O
authors.lastname: Hibbitt
authors.affiliation: Departments of Physiology, Anatomy, and Genetics (A.G.K., L.C.S.T., M.C., S.A., O.H., J.H.-M., R.W.-M.) and Pharmacology (A.J.R.), University of Oxford, Oxford, United Kingdom; Division of Cardiovascular Medicine, British Heart Foundation Centre of Research Excellence, University of Oxford, John Radcliffe Hospital, Oxford, United Kingdom (A.B.H., G.D., K.M.C.); and Chemistry Research Laboratory, Department of Chemistry, University of Oxford, Oxford, United Kingdom (J.M., C.J.R.B., G.M.W., A.J.R.).
authors.forename: Joseph
authors.initials: J
authors.lastname: Mason
authors.affiliation: Departments of Physiology, Anatomy, and Genetics (A.G.K., L.C.S.T., M.C., S.A., O.H., J.H.-M., R.W.-M.) and Pharmacology (A.J.R.), University of Oxford, Oxford, United Kingdom; Division of Cardiovascular Medicine, British Heart Foundation Centre of Research Excellence, University of Oxford, John Radcliffe Hospital, Oxford, United Kingdom (A.B.H., G.D., K.M.C.); and Chemistry Research Laboratory, Department of Chemistry, University of Oxford, Oxford, United Kingdom (J.M., C.J.R.B., G.M.W., A.J.R.).
authors.forename: James
authors.initials: J
authors.lastname: Holt-Martyn
authors.affiliation: Departments of Physiology, Anatomy, and Genetics (A.G.K., L.C.S.T., M.C., S.A., O.H., J.H.-M., R.W.-M.) and Pharmacology (A.J.R.), University of Oxford, Oxford, United Kingdom; Division of Cardiovascular Medicine, British Heart Foundation Centre of Research Excellence, University of Oxford, John Radcliffe Hospital, Oxford, United Kingdom (A.B.H., G.D., K.M.C.); and Chemistry Research Laboratory, Department of Chemistry, University of Oxford, Oxford, United Kingdom (J.M., C.J.R.B., G.M.W., A.J.R.).
authors.forename: Carole J R
authors.initials: CJR
authors.lastname: Bataille
authors.affiliation: Departments of Physiology, Anatomy, and Genetics (A.G.K., L.C.S.T., M.C., S.A., O.H., J.H.-M., R.W.-M.) and Pharmacology (A.J.R.), University of Oxford, Oxford, United Kingdom; Division of Cardiovascular Medicine, British Heart Foundation Centre of Research Excellence, University of Oxford, John Radcliffe Hospital, Oxford, United Kingdom (A.B.H., G.D., K.M.C.); and Chemistry Research Laboratory, Department of Chemistry, University of Oxford, Oxford, United Kingdom (J.M., C.J.R.B., G.M.W., A.J.R.).
authors.forename: Graham M
authors.initials: GM
authors.lastname: Wynne
authors.affiliation: Departments of Physiology, Anatomy, and Genetics (A.G.K., L.C.S.T., M.C., S.A., O.H., J.H.-M., R.W.-M.) and Pharmacology (A.J.R.), University of Oxford, Oxford, United Kingdom; Division of Cardiovascular Medicine, British Heart Foundation Centre of Research Excellence, University of Oxford, John Radcliffe Hospital, Oxford, United Kingdom (A.B.H., G.D., K.M.C.); and Chemistry Research Laboratory, Department of Chemistry, University of Oxford, Oxford, United Kingdom (J.M., C.J.R.B., G.M.W., A.J.R.).
authors.forename: Keith M
authors.initials: KM
authors.lastname: Channon
authors.affiliation: Departments of Physiology, Anatomy, and Genetics (A.G.K., L.C.S.T., M.C., S.A., O.H., J.H.-M., R.W.-M.) and Pharmacology (A.J.R.), University of Oxford, Oxford, United Kingdom; Division of Cardiovascular Medicine, British Heart Foundation Centre of Research Excellence, University of Oxford, John Radcliffe Hospital, Oxford, United Kingdom (A.B.H., G.D., K.M.C.); and Chemistry Research Laboratory, Department of Chemistry, University of Oxford, Oxford, United Kingdom (J.M., C.J.R.B., G.M.W., A.J.R.).
authors.forename: Angela J
authors.initials: AJ
authors.lastname: Russell
authors.affiliation: Departments of Physiology, Anatomy, and Genetics (A.G.K., L.C.S.T., M.C., S.A., O.H., J.H.-M., R.W.-M.) and Pharmacology (A.J.R.), University of Oxford, Oxford, United Kingdom; Division of Cardiovascular Medicine, British Heart Foundation Centre of Research Excellence, University of Oxford, John Radcliffe Hospital, Oxford, United Kingdom (A.B.H., G.D., K.M.C.); and Chemistry Research Laboratory, Department of Chemistry, University of Oxford, Oxford, United Kingdom (J.M., C.J.R.B., G.M.W., A.J.R.) richard.wade-martins@dpag.ox.ac.uk angela.russell@chem.ox.ac.uk.
authors.forename: Richard
authors.initials: R
authors.lastname: Wade-Martins
mesh_terms: D000818:Animals; D016466:CHO Cells; D045744:Cell Line, Tumor; D006224:Cricetinae; D003412:Cricetulus; D004305:Dose-Response Relationship, Drug; D004357:Drug Synergism; D004791:Enzyme Inhibitors; D013186:Farnesyl-Diphosphate Farnesyltransferase / Q000037:antagonists & inhibitors* / Q000378:metabolism; D005820:Genetic Testing / Q000379:methods*; D006801:Humans; D019161:Hydroxymethylglutaryl-CoA Reductase Inhibitors / Q000008:administration & dosage*; D008297:Male; D051379:Mice; D011973:Receptors, LDL / Q000096:biosynthesis*; D054852:Small Molecule Libraries / Q000008:administration & dosage* / Q000494:pharmacology; D015854:Up-Regulation / Q000187:drug effects / Q000502:physiology*
chemical_list: D004791:Enzyme Inhibitors; D019161:Hydroxymethylglutaryl-CoA Reductase Inhibitors; C573412:LDLR protein, human; D011973:Receptors, LDL; D054852:Small Molecule Libraries; D013186:Farnesyl-Diphosphate Farnesyltransferase