title: Differential in vivo effects on target pathways of a novel arylpyrazole glucocorticoid receptor modulator compared with prednisolone.
abstract: Glucocorticoids are widely prescribed to treat autoimmune and inflammatory diseases. Although they are extremely potent, their utility in clinical practice is limited by a variety of adverse side effects. Development of compounds that retain the potent immunomodulating and anti-inflammatory properties of classic glucocorticoids while exhibiting reduced adverse actions is therefore a priority. Using heavy water labeling and mass spectrometry to measure fluxes through multiple glucocorticoid-responsive, disease-relevant target pathways in vivo in mice, we compared the effects of a classic glucocorticoid receptor (GR) ligand, prednisolone, with those of a novel arylpyrazole-based compound, L5 {[1-(4-fluorophenyl)-4a-methyl-5,6,7,8-tetrahydro-4H-benzo[f]indazol-5-yl]-[4-(trifluoromethyl)phenyl]methanol}. We show for the first time that L5 exhibits clearly selective actions on disease-relevant pathways compared with prednisolone. Prednisolone reduced bone collagen synthesis, skin collagen synthesis, muscle protein synthesis, and splenic lymphocyte counts, proliferation, and cell death, whereas L5 had none of those actions. In contrast, L5 was a more rapid and potent inhibitor of hippocampal neurogenesis than prednisolone, and L5 and prednisolone induced insulin resistance equally. Administration of prednisolone or L5 increased expression comparably for one GR-regulated gene involved in protein degradation in skeletal muscle (Murf1) and one GR-regulated gluconeogenic gene in liver (PEPCK). In summary, L5 dissociates the pleiotropic effects of the GR ligand prednisolone in intact animals in ways that neither gene expression nor cell-based models were able to fully capture or predict. Because multiple actions can be measured concurrently in a single animal, this method is a powerful systems approach for characterizing and differentiating the effects of ligands that bind nuclear receptors.
authors.affiliation: Department of Nutritional Sciences and Toxicology, 309 Morgan Hall, University of California, Berkeley, CA 94720-3104, USA. djroohk@berkeley.edu
authors.forename: Donald J
authors.initials: DJ
authors.lastname: Roohk
authors.forename: Krista A
authors.initials: KA
authors.lastname: Varady
authors.forename: Scott M
authors.initials: SM
authors.lastname: Turner
authors.forename: Claire L
authors.initials: CL
authors.lastname: Emson
authors.forename: Richard W
authors.initials: RW
authors.lastname: Gelling
authors.forename: Mahalakshmi
authors.initials: M
authors.lastname: Shankaran
authors.forename: Glen
authors.initials: G
authors.lastname: Lindwall
authors.forename: Lauren E
authors.initials: LE
authors.lastname: Shipp
authors.forename: Thomas S
authors.initials: TS
authors.lastname: Scanlan
authors.forename: Jen-Chywan
authors.initials: JC
authors.lastname: Wang
authors.forename: Marc K
authors.initials: MK
authors.lastname: Hellerstein
mesh_terms: D000273:Adipose Tissue / Q000187:drug effects / Q000378:metabolism; D000818:Animals; D001842:Bone and Bones / Q000187:drug effects / Q000378:metabolism; D016923:Cell Death / Q000187:drug effects; D049109:Cell Proliferation / Q000187:drug effects; D003094:Collagen / Q000096:biosynthesis; D020869:Gene Expression Profiling; D005938:Glucocorticoids / Q000494:pharmacology*; D006624:Hippocampus / Q000166:cytology / Q000187:drug effects; D007191:Indazoles / Q000494:pharmacology*; D007333:Insulin Resistance; D007515:Islets of Langerhans / Q000166:cytology / Q000187:drug effects; D008099:Liver / Q000187:drug effects / Q000378:metabolism; D018655:Lymphocyte Count; D008297:Male; D051379:Mice; D008810:Mice, Inbred C57BL; D018482:Muscle, Skeletal / Q000187:drug effects / Q000378:metabolism; D055495:Neurogenesis / Q000187:drug effects; D011239:Prednisolone / Q000494:pharmacology*; D014176:Protein Biosynthesis / Q000187:drug effects; D011965:Receptors, Glucocorticoid / Q000502:physiology*; D015398:Signal Transduction / Q000187:drug effects*; D012867:Skin / Q000187:drug effects / Q000378:metabolism; D013154:Spleen / Q000166:cytology / Q000187:drug effects; D013234:Stem Cells / Q000187:drug effects; D014280:Triglycerides / Q000378:metabolism
chemical_list: C548783:(1-(4-fluorophenyl)-4a-methyl-5,6,7,8-tetrahydro-4H-benzo(f)indazol-5-yl)-(4-(trifluoromethyl)phenyl)methanol; D005938:Glucocorticoids; D007191:Indazoles; D011965:Receptors, Glucocorticoid; D014280:Triglycerides; D003094:Collagen; D011239:Prednisolone