Late stage assay provider counterscreen results from the probe development effort to identify non-agonists of the peroxisome proliferator-activated receptor gamma (PPARg): Leptin-deficient ob/ob mouse model studies to assess the effect of probe candidate on fasting blood glucose
Name:Late stage assay provider counterscreen results from the probe development effort to identify non-agonists of the peroxisome proliferator-activated receptor gamma (PPARg): Leptin-deficient ob/ob mouse model studies to assess the effect of probe candidate on fasting blood glucose. ..more
Depositor Specified Assays
Source (MLPCN Center Name): The Scripps Research Institute Molecular Screening Center (SRISMC)
Center Affiliation: The Scripps Research Institute, TSRI
Assay Provider: Patrick Griffin, TSRI
Network: Molecular Library Probe Production Center Network (MLPCN)
Grant Proposal Number: U54 MH084512
Grant Proposal PI: Patrick Griffin, TSRI
External Assay ID: LEPTIN-DEFICIENT-OB-OB-MOUSE_GLUCOSE_INH NON-AG PPARG
Name:Late stage assay provider counterscreen results from the probe development effort to identify non-agonists of the peroxisome proliferator-activated receptor gamma (PPARg): Leptin-deficient ob/ob mouse model studies to assess the effect of probe candidate on fasting blood glucose.
Peroxisome proliferator-activated receptors (PPARs) belong to the nuclear receptor superfamily and are lipid sensors functioning as ligand-dependent transcription factors regulating gene expression patterns of diverse biological processes (1, 2). PPARs play a critical role in metabolic processes such as glucose metabolism, lipid metabolism, and have been implicated in anti-atherogenic, anti-inflammatory as well as anti-hypertensive functions (3). Like other nuclear receptors, PPARs act as agonist-activated transcription factors, regulating specific PPARG gene transcription. PPARs have been shown to respond to small molecules and are well-documented for therapeutic actions triggered by synthetic agonists (4-6). Among the three isoforms of PPAR identified, PPAR gamma (NR1C3) is implicated in several important disorders such as atherosclerosis, diabetes, obesity and cancer, providing strong justification for the search for specific PPARg agonists that can be used to treat these pathologies. However, the clinical use of PPARg agonists has been associated with adverse effects that are mainly caused by the concomitant activation of various PPARG genes implicated in different physiological pathways. These side effects include weight gain through increased adipogenesis, renal fluid retention and plasma volume expansion, as well as toxic effects in the liver (7). To design safer and more selective PPARg agonists, the different physiological pathways triggered by PPARg activation have to be decoupled. Recently, new classes of PPARg ligands, the so called selective PPARg modulators (SPPARgMs), have been developed. These compounds respond as partial agonists in a GAL-4 luciferase assay and are assumed to display a different binding mode in the PPARg subunit compared to the full agonist, glitazones (8). Selective recruitment of transcriptional coactivators by partial agonists has also been demonstrated, suggesting that different PPARg binding mode leading to a distinct coactivator recruitment profile may explain the change in gene expression patterns compared to those of full agonists (glitazones). Further, due to their improved pharmacodynamic properties, there is substantial interest and need to develop insulin-sensitizing PPARg modulators with minimal classical activation of PPARg and reduced side effects, while maintaining robust antidiabetic efficacy (9-11). The objective of this project is to identify compounds that bind to PPARg and do not induce PPARg transactivation (ligands; non-agonists) (12).
1. Chawla, A., et al., Nuclear receptors and lipid physiology: Opening the X-files. Science, 2001. 294(5548): p. 1866-1870.
2. Krey, G., et al., Fatty acids, eicosanoids, and hypolipidemic agents identified as ligands of peroxisome proliferator-activated receptors by coactivator-dependent receptor ligand assay. Molecular Endocrinology, 1997. 11(6): p. 779-791.
3. Bishop-Bailey, D., T. Hla, and T.D. Warner, Intimal smooth muscle cells as a PPARG for peroxisome proliferator-activated receptor-gamma ligand therapy. Circ Res, 2002. 91(3): p. 210-7.
4. Evans, R.M., G.D. Barish, and Y.X. Wang, PPARs and the complex journey to obesity. Nat Med, 2004. 10(4): p. 355-61.
5. Staels, B., et al., Mechanism of action of fibrates on lipid and lipoprotein metabolism. Circulation, 1998. 98(19): p. 2088-93.
6. Barish, G.D., V.A. Narkar, and R.M. Evans, PPAR delta: a dagger in the heart of the metabolic syndrome. J Clin Invest, 2006. 116(3): p. 590-7.
7. Berger, J.P., T.E. Akiyama, and P.T. Meinke, PPARs: therapeutic PPARGs for metabolic disease. Trends Pharmacol Sci, 2005. 26(5): p. 244-51.
8. Berger J, Leibowitz MD, Doebber TW, Elbrecht A, Zhang B, Zhou G, Biswas C, Cullinan CA, Hayes NS, Li Y, Tanen M, Ventre J, Wu MS, Berger GD, Mosley R, Marquis R, Santini C, Sahoo SP, Tolman RL, Smith RG, Moller DE. Novel peroxisome proliferator-activated receptor (PPAR) gamma and PPARdelta ligands produce distinct biological effects. J Biol Chem. 1999 Mar 5;274(10):6718-25.
9. Berger JP, Petro AE, Macnaul KL, Kelly LJ, Zhang BB, Richards K, Elbrecht A, Johnson BA, Zhou G, Doebber TW, Biswas C, Parikh M, Sharma N, Tanen MR, Thompson GM, Ventre J, Adams AD, Mosley R, Surwit RS, Moller DE.Distinct properties and advantages of a novel peroxisome proliferator-activated protein [gamma] selective modulator. Mol Endocrinol. 2003 Apr;17(4):662-76.
10. Minoura H, Takeshita S, Ita M, Hirosumi J, Mabuchi M, Kawamura I, Nakajima S, Nakayama O, Kayakiri H, Oku T, Ohkubo-Suzuki A, Fukagawa M, Kojo H, Hanioka K, Yamasaki N, Imoto T, Kobayashi Y, Mutoh S.
Eur J Pharmacol. 2004 Jun 28;494(2-3):273-81. Pharmacological characteristics of a novel nonthiazolidinedione insulin sensitizer, FK614.
11. Vidovic D, Busby SA, Griffin PR, Schurer SC. A combined ligand- and structure-based virtual screening protocol identifies submicromolar PPARg partial agonists. ChemMedChem. 2011 Jan 3;6(1):94-103.
12. Choi JH, Banks AS, Estall JL, Kajimura S, Bostrom P, Laznik D, Ruas JL, Chalmers MJ, Kamenecka TM, Bluher M, Griffin PR, Spiegelman BM. Anti-diabetic drugs inhibit obesity-linked phosphorylation of PPARgamma by Cdk5. Nature. 2010 Jul 22;466(7305):451-6.
mouse, leptin, obese, mouse, fasting, ob, diabetes, glucose, metabolism, probe, counterscreen, PPARG, Late stage, late stage AID, powders, purchased, synthesized, PPAR gamma, PPARg, PPARG1, PPARG2, PPAR, peroxisome proliferator-activated receptor gamma, partial agonist, agonist, non-agonist, ligand, inhibit, assay provider, CBI, center based initiative, center-based, biochemical, selective, nuclear receptor, tumor, cancer, dose response, triplicate, Scripps Florida, The Scripps Research Institute Molecular Screening Center, SRIMSC, Molecular Libraries Probe Production Centers Network, MLPCN.
§ Panel component ID.
The purpose of this assay is to determine whether a powder sample of a compound identified as a novel PPARG ligand and non-agonist probe candidate (ML244) can improve glucose tolerance and thereby reduce blood glucose levels in the leptin deficient ob/ob mouse model. The probe should be active in this assay. The compound was tested in animals at 40 mg/kg. In this assay the control was vehicle (not active) and the positive control was rosiglitazone. Rosiglitazone and the probe ML244 each induced significant reductions in blood glucose levels at the timepoints tested.
All animal experiments were performed according to procedures approved by Beth Israel Deaconess Medical Center's Institutional Animal Care and Use Committee. 4 to 5 week-old male C57BL/6J and C57BL/6J-Lepob/ob mice were obtained from the Jackson Laboratory. C57BL/6J mice were fed a high fat, high sucrose diet (60% kcal fat, D12492, Research Diets Inc.). For glucose tolerance tests, mice were intraperitoneally (i.p.) injected with rosiglitazone or probe compound (ML244; SR1664) for 5 days, and fasted overnight before i.p. injection of 1 g/kg D-glucose.
The high control reference compound, Rosiglitazone, led to the following blood glucose levels at the indicated timepoints following glucose injection:
0 min, 100 mg/dl
30 min, 300 mg/dl
60 min, 250 mg/dl
90 min, 190 mg/dl
120 min, 150 mg/dl
Animals treated with vehicle alone (low control, inactive) had he following blood glucose levels at the indicated timepoints following glucose injection:
0 m, 100 mg/dl
30 min, 450 mg/dl
60 min, 420 mg/dl
90 min, 400 mg/dl
120 min, 350 mg/dl
PubChem Activity Outcome and Score:
Compounds that decreased blood glucose to levels comparable to that of animals treated with rosiglitazone were considered active. The probe was active.
The PubChem Activity Score is assigned a value of 100 for probe compounds, 50 for actives and 0 for inactives.
The PubChem Activity Score range for active compounds is 100-100. There were no inactive compounds.
List of Reagents:
See protocol text.
This assay was run by the assay provider's collaborator. This assay may have been run as two or more separate campaigns, each campaign testing a unique set of compounds. All test compound concentrations reported above and below are nominal; the specific test concentration(s) for a particular compound may vary based upon the actual sample provided.