GW-6471 - PPAR Antagonist - MedChemExpress

1、Hotline: 400-820-3792Inhibitors Agonists Screening Librarieswww.MedChemEGW 6471Cat. No.: HY-15372CAS No.: 880635-03-0分式: CHFNO分量: 619.67作靶点: PPAR作通路: Cell Cycle/DNA Damage储存式: Powder -20C 3 years4C 2 yearsIn solvent -80C 6 months-20C 1 month溶解性数据体外实验 DMSO : 125 mg/mL (201.72 mM; Need ultrasonic)H2O

2、: 40% PEG300 5% Tween-80 45% salineSolubility: 2.08 mg/mL (3.36 mM); Clear solution2. 请依序添加每种溶剂： 10% DMSO 90% corn oilSolubility: 2.08 mg/mL (3.36 mM); Clear solution1/3 Master of Small Molecules 您边的抑制剂师www.MedChemEBIOLOGICAL ACTIVITY物活性 GW 6471种有效的 PPAR 拮抗剂。IC50 & Target PPAR体外研究 In a cell-based re

3、porter assay, GW 6471 completely inhibits GW409544-induced activation of PPAR with anIC50 of 0.24 M 1. The functional role of PPAR is evaluated on renal cell carcinoma (RCC) cell viability byMTT assay. Both Caki-1 (VHL wild type) and 786-O (VHL mutated) cells are incubated separately with aspecific

4、PPAR agonist, WY14,643, or a specific PPAR antagonist, GW 6471 at concentrations from 12.5 to100 M for 72 hours, and cell viability is assessed. While WY14,643 either has no affect on, or slightlyincreased, cell viability, GW 6471 significantly and dose-dependently inhibits cell viability (up toappr

5、oximately 80%) in both cell lines 2.体内研究 To test the antitumor activity of PPAR antagonism in vivo, a subcutaneous xenograft mouse model is used.Caki-1 cells are implanted subcutaneously in nude (Nu/Nu) mice. After tumor masses reach -5 mm indiameter, GW 6471 is administrated intraperitoneally every

6、 other day for 4 wk at a dose (20 mg/kg mousebody wt) that is described to be effective in an in vivo dose-response study and confirmed here to beefficacious. There are significant differences in tumor growth between vehicle- and GW 6471-treatedanimals. No toxicity is observed at the doses of GW 647

7、1 based on weights of the animals, and laboratoryvalues, including kidney and liver function tests, are not adversely affected. To demonstrate on-target effectsof GW 6471, c-Myc levels are evaluated in the tumors, which show significant decreases in the GW 6471-treated animals 3.PROTOCOLCell Assay 2

8、 786-O and Caki-1 cells are plated in 96 well plates. Both cells are incubated separately with WY14,643 orGW 6471 at concentrations from 12.5 to 100 M for 72 hours, and after the indicated treatments, the cellsare incubated in MTT solution/media mixture. Then, the MTT solution is removed and the blu

9、e crystallineprecipitate in each well is dissolved in DMSO. Visible absorbance of each well at 540 nm is quantified using amicroplate reader 2.MCE has not independently confirmed the accuracy of these methods. They are for reference only.Animal Mice 3Administration 3 Male athymic Nu/Nu mice (8 wk of

10、 age, 25 g body wt) are injected with 1105 Caki-1 cells subcutaneously(3:1 DMEM-Matrigel) in the flank region. Tumor progression is monitored weekly by calipers. When tumorsize reaches 80-100 mm3, animals are randomly assigned to four groups and treatments are started (day1). The vehicle group recei

11、ve DMSO (4% in PBS) intraperitoneally and vegetable oil via oral gavage. ThePPAR group is injected intraperitoneally with GW 6471 in the same vehicle (20 mg/kg body wt; murine doseresponse is reported elsewhere) every other day. The Sunitinib group receive Sunitinib in vegetable oil viaoral gavage (

12、40 mg/kg body wt) 5 days/wk. Another group receive GW 6471+Sunitinib. To determine anypotential toxicity of the treatment(s), body weights of the animals are measured and signs of adversereactions are monitored. On day 28, the mice are euthanized and the tumor mass is determined. Tumorgrowth rate is

13、 calculated 3.2/3 Master of Small Molecules 您边的抑制剂师www.MedChemEMCE has not independently confirmed the accuracy of these methods. They are for reference only.REFERENCES1. Xu HE, et al. Structural basis for antagonist-mediated recruitment of nuclear co-repressors by PPARalpha. Nature. 2002 Feb14;415(

14、6873):813-7.2. Abu Aboud O, et al. Inhibition of PPAR induces cell cycle arrest and apoptosis, and synergizes with glycolysisinhibition in kidneycancer cells. PLoS One. 2013 Aug 7;8(8):e71115.3. Abu Aboud O, et al. PPAR inhibition modulates multiple reprogrammed metabolic pathways in kidney cancer and attenuates tumorgrowth. Am J Physiol Cell Physiol. 2015 Jun 1;308