A-966492 _PARP 抑制剂 - MedChemExpress

1、Hotline: 400-820-3792Inhibitors Agonists Screening Librarieswww.MedChemEA-966492Cat. No.: HY-10614CAS No.: 934162-61-5分式: CHFNO分量: 324.35作靶点: PARP作通路: Cell Cycle/DNA Damage; Epigenetics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 : 100 mg/mL (308.31 mM)* means

2、 soluble, but saturation unknown.Mass Solvent1 mg 5 mg 10 mg Concentration制备储备液1 mM 3.0831 mL 15.4154 mL 30.8309 mL5 mM 0.6166 mL 3.0831 mL 6.1662 mL10 mM 0.3083 mL 1.5415 mL 3.0831 mL请根据产品在不同溶剂中的溶解度，选择合适的溶剂配制储备液，并请注意储备液的保存式和期限。体内实验请根据您的实验动物和给药式选择适当的溶解案，配制前请先配制澄清的储备液，再依次添加助溶剂(为保证实验结果的可靠性，体内实验的作液，建议您

3、现现配，当天使；澄清的储备液可以根据储存条件，适当保存；以下溶剂前的百分 指该溶剂在您配制终溶液中的体积占)：1. 请依序添加每种溶剂： 10% DMSO 40% PEG300 5% Tween-80 45% salineSolubility: 2.5 mg/mL (7.71 mM); Clear solution2. 请依序添加每种溶剂： 10% DMSO 90% (20% SBE-CD in saline)Solubility: 2.5 mg/mL (7.71 mM); Clear solution3. 请依序添加每种溶剂： 10% DMSO 90% corn oil1/3 Master

4、of Small Molecules 您边的抑制剂师www.MedChemESolubility: 2.5 mg/mL (7.71 mM); Clear solutionBIOLOGICAL ACTIVITY物活性 A-966492种新颖的，有效的 PARP1 和 PARP2 抑制剂，Ki 值为 1 nM 和 1.5 nM。IC50 & Target PARP-1 PARP-21 nM (Ki) 1.5 nM (Ki)体外研究 A-966492 is one of the most potent PARP inhibitors. A-966492 displays excellent pote

5、ncy against the PARP-1 enzyme with a Ki of 1 nM and an EC50 of 1 nM in a whole cell assay. A-966492 significantly enhances theefficacy of TMZ in a dose-dependent manner. In addition, A-966492 is orally bioavailable across multiplespecies, crosses the bloodbrain barrier, and appears to distribute int

6、o tumor tissue. A-966492 represents apromising, structurally diverse benzimidazole analogue and is being further characterized preclinically 1.体内研究 A-966492 demonstrates good in vivo efficacy in a B16F10 subcutaneous murine melanoma model incombination with temozolomide and in an MX-1 breast cancer

7、xenograft model both as a single agent and incombination with carboplatin. In addition, A-966492 has excellent pharmaceutical properties and hasdemonstrated in vivo efficacy in preclinical mouse tumor models in combination with TMZ and carboplatin, aswell as single agent activity in a BRCA1-deficien

8、t MX-1 tumor model. A-966492 is further characterized inSpragueDawley rats, beagle dogs, and cynomolgus monkeys, with A-966492 demonstrating oralbioavailabilities of 3472% and half-lives of 1.71.9 hours. In vivo, A-966492 demonstrates significantenhancement of the efficacy of TMZ in a murine B16F10

9、syngeneic melanoma model, with the A-966492combination groups showing superior efficacy 1.PROTOCOLKinase Assay 1 The enzyme assay is conducted in buffer containing 50 mM Tris, pH 8.0, 1 mM dithiothreitol(DTT), and 4 mMMgCl2. PARP reactions contains 1.5 M 3H-NAD+ (1.6 Ci/mmol), 200 nM biotinylated hi

10、stone H1, 200 nMslDNA, and 1 nM PARP-1 or 4 nM PARP-2 enzyme. Autoreactions utilizing SPA bead-based detection arecarried out in 100 L volumes in white 96-well plates. Reactions are initiated by adding 50 L of 2X NAD+substrate mixture to 50 L of 2 enzyme mixture containing PARP and DNA. These reacti

11、ons are terminatedby the addition of 150 L of 1.5 mM benzamide (appr 1103-fold over its IC50). A 170 L amount of thestopped reaction mixtures is transferred to streptavidin-coated Flash Plates, incubated for 1 hour, andcounted using a TopCount microplate scintillation counter. Ki data are determined

12、 from inhibition curves atvarious substrate concentrations.MCE has not independently confirmed the accuracy of these methods. They are for reference only.Cell Assay 1 C41 cells are treated with A-966492 for 30 minutes in a 96-well plate. PARP are activated by damaging DNAwith 1 mM H2O2 for 10 minute

13、s. Cells are washed with ice-cold phosphate-buffered saline (PBS) once andfixed with prechilled methanol/acetone (7:3) at -20C for 10 minutes. After they are air-dried, plates arerehydrated with PBS and blocked using 5% nonfat dry milk in PBS-Tween (0.05%) (blocking solution) for 302/3 Master of Sma

14、ll Molecules 您边的抑制剂师www.MedChemEminutes at room temperature. Cells are incubated with anti-PAR antibody 10H (1:50) in blocking solution atroom temperature for 60 minutes followed by washing with PBS-Tween20 five times, and incubation withgoat antimouse fluorescein 5(6)-isothiocyanate (FITC)-coupled

15、antibody (1:50) and 1 g/mL 40,6-diamidino-2-phenylindole (DAPI) in blocking solution at room temperature for 60 minutes. After washing with PBS-Tween20 5 times, analysis is performed using an fmax Fluorescence Microplate Reader set at the excitationand emission wavelength for FITC or the excitation

16、and emission wavelength for DAPI. PARP activity (FITCsignal) is normalized with cell numbers (DAPI).MCE has not independently confirmed the accuracy of these methods. They are for reference only.Animal A 0.2 cc amount of a 1:10 dilution of tumor brei in 45% Matrigel and 45% Spinner MEM is injectedAd

17、ministration 1 subcutaneously into the flank of female SCID mice on study day 0. Tumors are allowed to grow to theindicated size and then randomized to therapy groups (N=10 mice/group). PARP inhibitor therapy begin onday 14, with cisplatin treatment starting on day 16. At various intervals following

18、 tumor inoculation, theindividual tumor dimensions are serially measured using calibrated microcalipers, and the tumor volumes arecalculated.MCE has not independently confirmed the accuracy of these methods. They are for reference only.户使本产品发表的科研献 J Mol Med (Berl). 2019 Jun 14. Patent. US20180263995A1.See more customer validations on HYPERLINK / www.MedChemEREFERENCES1. Penning TD, et al. Optimization of phenyl-substituted benzimidazole carboxamide poly(ADP-ribose) polymerase inhibitors: identificationof (S)-2-(2-fluoro-4-(pyrrolidin-2-y