Cort108297 - Glucocorticoid Receptor Antagonist - MedChemExpress

1、Hotline: 400-820-3792Inhibitors Agonists Screening Librarieswww.MedChemECort108297Cat. No.: HY-15710CAS No.: 1018679-79-2分式: CHFNOS分量: 535.55作靶點: Glucocorticoid Receptor作通路: GPCR/G Protein儲存式: Powder -20C 3 years4C 2 yearsIn solvent -80C 6 months-20C 1 monthBIOLOGICAL ACTIVITY物活性 Cort108297種親和性的特異性糖

2、質(zhì)激素受體 (GR) 拮抗劑，Ki 值為 0.45 nM。IC50 & Target Ki: 0.45 nM (glucocorticoid receptor) 1體外研究 In LAPC4 cells, co-treatment with Dexamethasone induces steady-state SGK1 expression 1.7-fold comparedto R1881/Enzalutamide (RE) treatment alone. Addition of CORT118335 (1M) inhibits Dexamethasone-induced SGK1 exp

3、ression 50% while Cort108297 completely blocks the Dexamethasone-mediated SGK1increase (pKLK3 expression is increased 2.5-fold by Dexamethasone compared to treatment with RE. BothCort108297 and CORT118335 antagonize Dexamethasone-induced KLK3 expression (by 48% and 60%,respectively, pSGK1 gene expre

4、ssion is dramatically induced by 100-fold compared to RE-treated cells andthis induction is completely abrogated by both Cort108297 and CORT118335 (pKLK3 is also induced (7.5-fold) by Dexamethasone compared to RE in CWR-22Rv1 cells; Cort108297 and CORT118335 inhibits thisinduction by 70% and 75%, re

5、spectively (p 2.體內(nèi)研究Ten-week-old, male, C57BL/6J mice are fed a diet containing 60% fat calories and water supplemented with11% sucrose for 4 weeks. Groups (n=8) receive one of the following: Cort108297 (80mg/kg QD),Cort108297 (40mg/kg BID), Mifepristone (30mg/kg BID), Rosiglitazone (10mg/kg QD), or

6、 vehicle.Compared to mice receiving a high-fat, high-sugar diet plus vehicle, mice receiving a high-fat, high-sugar dietplus either Mifepristone or Cort108297 gain significantly less weight. At the end of the four week treatmentperiod, mice receiving Cort108297 40mg/kg BID or Cort108297 80mg/kg QD a

7、lso have significantly lowersteady plasma glucose than mice receiving vehicle 3. Male rats are treated for five days with Mifepristone1/3 Master of Small Molecules 您邊的抑制劑師www.MedChemE(10 mg/kg), Cort108297 (30 mg/kg and 60 mg/kg), Imipramine (10mg/kg) or vehicle and exposed to forcedswim test (FST)

8、or restraint stress. Both doses of Cort108297 potently suppress peak corticosteroneresponses to FST and restraint stress. However, only the higher dose of Cort108297 (60mg/kg) significantlydecreases immobility in the forced swim test (FST) 4.PROTOCOLKinase Assay 1 Steroid receptor competition bindin

9、g assays are run in a buffer containing 20 mM HEPES buffer (pH=7.6),0.2 mM EDTA, 75 mM NaCl, 1.5 mM MgCl2, 20% glycerol, 20 mM sodium molybdate, 0.2 mM DTT, 20 g/mL Aprotinin, and 20 g/mL Leupeptin (assay buffer). Radiolabeled ligands are used to detect binding tocells expressing receptors including

10、 0.25 nM 3HAldosterone for mineralocorticoid receptor (MR) binding, 0.3nM 3HDexamethasone for GR binding, 0.36 nM 3HMethyltrienolone for aldosterone receptor (AR) binding,and 0.29 nM 3Hmethyltrienolone for PR binding. Receptors are recombinantly expressed in humanembryonic kidney 293 (HEK-293) cells

11、, and 20 g of 293-MR lysate, 20 g of 293-GR lysate, 22 g of 293-AR lysate, or 40 g of 293-PR lysate are added per well. Competing test compounds (e.g., Cort108297) areadded at various concentrations from 0.01 nM to 10 M. Nonspecific binding is determined in the presence of500 nM Aldosterone for MR b

12、inding, 500 nM Dexamethasone for GR binding, or 500 nM methyltrienolone forAR and PR binding. The binding reactions (140 L) are incubated overnight at 4C, then 70 l of coldcharcoal-dextran buffer (containing per 50 mL of assay buffer, 0.75 g of Charcoal, and 0.25 g of Dextran) isadded to each reacti

13、on. Plates are mixed for 8 minutes on an orbital shaker at 4C. The plates are thencentrifuged at 3000 rpm at 4C for 10 minutes. A 120 L aliquot of the binding reaction mixture is thentransferred to another 96-well plate, and 175 L of Wallac Optiphase Hisafe 3 scintillation fluid is added toeach well

14、. The plates are sealed and shaken vigorously using an orbital shaker. After 2 hour incubation, theplates are counted using a Wallac MicroBeta counter 1.MCE has not independently confirmed the accuracy of these methods. They are for reference only.Cell Assay 2 LAPC4 and CWR-22Rv1 cells are plated in

15、 standard media and incubated overnight. Cells are washed withPBS and placed into media containing charcoal stripped FBS, 10% for LAPC4 or 1%/10% for CWR-22Rv1.Cells are treated for indicated times with media changes every other day with either vehicle control orspecified treatment: 1 nM R1881, 100

16、nM Dexamethasone, 10 M Enzalutamide, 100 nM Mifepristone, 1 MCORT118335, 1 M Cort108297. For all experiments, equimolar vehicle (ethanolDMSO) is added to everysample for equal treatment periods. Cells are plated and treated. At indicated days cells are washed,trypsinized, pelleted, and resuspended i

17、n media. Cells are then mixed 1:1 with trypan blue and viable cellsare counted in a blinded fashion. Three biological replicates are assayed per condition per time point and themean of the biological replicates is reported 2.MCE has not independently confirmed the accuracy of these methods. They are

18、 for reference only.Animal Mice 3Administration 34 Forty ten-week-old, male, C57BL/6J mice are fed ad libitum a diet containing 60% fat calories and watersupplemented with 11% sucrose for 4 weeks. In addition, they receive one of the following five treatments:Cort108297 (80mg/kg QD), Cort108297 (40m

19、g/kg BID), Mifepristone (30mg/kg BID), Rosiglitazone, an oralglycemic medication (10mg/kg QD), or vehicle (10% DMSO in 0.5% CMC). An additional control group (n=8)is fed a standard chow diet and tap water and does not receive any treatment.2/3 Master of Small Molecules 您邊的抑制劑師www.MedChemERats 4Male

20、Sprague Dawley rats (250-275 g) are used. Forty-eight rats are matched by body weight and areadministered, Cort108297 dissolved in DMSO (30mg/kg s.c.(n=10) or 60 mg/kg s.c. (n=10), Mifepristonedissolved in DMSO 10mg/kg s.c. (n=10), Imipramine dissolved in saline 10mg/kg i.p. (n=10) or vehicle DMSOs.

21、c. (n=4) or saline i.p. (n=4). Control groups consist of both subcutaneous (s.c.) and intraperitoneal (i.p.)groups to control for the route of administration and both DMSO and saline to control for any potentialdifferences between the compounds on neuroendocrine and behavioral stress responsiveness.MCE has not independently confirmed the accuracy of these methods. They are for reference only.REFERENCES1. Sindelar DK, et al. LLY-2707, a novel nonsteroidal glucocorticoid antagonist that reduces atypical antipsychotic-associated weight gainin rats. J Pharmacol Exp Ther. 201