PF-9366-DataSheet-生命科学试剂-MedChemExpress

1、Hotline: 400-820-3792Inhibitors Agonists Screening Librarieswww.MedChemEPF-9366Cat. No.: HY-107778CAS No.: 72882-78-1分式: CHClN分量: 350.84作靶点: Others作通路: Others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 : 25 mg/mL (71.26 mM; Need ultrasonic)Mass Solvent1 mg 5

2、mg 10 mg Concentration制备储备液1 mM 2.8503 mL 14.2515 mL 28.5030 mL5 mM 0.5701 mL 2.8503 mL 5.7006 mL10 mM 0.2850 mL 1.4252 mL 2.8503 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

3、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 (7.13 mM); Clear solution2. 请依序添加每种溶剂： 10% DMSO 90% corn oilSolubility: 2.5 mg/mL (7.13 mM); Clear solutionBIOLOGICAL ACTIVITY1/3 Master of Small Molecules 您边的抑制剂师www.MedChemE物活性 PF-9366种有效的甲硫氨酸腺苷转移酶 (Mat2A) 抑制剂，IC50 值

4、和 Kd 值分别为 420 nM 和 170 nM。IC50 & Target IC50: 420 nM (Mat2A) 1Kd: 170 nM (Mat2A) 1体外研究 PF-9366 is a Mat2A inhibitor, with an IC50 of 420 nM and a Kd of 170 nM. PF-9366 displays no substantialoff-target activity in GPCRs, neurotransporters, phosphodiesterases, and ion channels. PF-9366 hasinhibitory

5、activity against Mat2A in cancer cells. PF-9366 inhibits cellular S-Adenosyl-L-methionine (SAM)production with an IC50 of 1.2 M in H520 lung carcinoma cells. PF-9366 is more potent in Huh-7 cellsagainst SAM synthesis, with an IC50 of 255 nM, and also suppresses the proliferation of cells with an IC5

6、0 of10 M.PROTOCOLKinase Assay 1 The Mat2A and Mat2B proteins are extensively dialyzed into a buffer containing 150 mM KCl, 25 mMHEPES, pH 7.4, 5 mM MgCl2, 5% (v/v) glycerol, 2 mM TCEP. Concentrations are determinedspectrophotometrically using an 280 of 44,350 /M.cm for Mat2A and an 280 of 36,440 /M.

7、cm for Mat2B.Compounds (PF-9366) are diluted from 100% DMSO stocks into a buffer without DMSO. In a typicalexperiment, nineteen 15 L injections of 200 M compound or 30-35 M Mat2B are made into 10 M Mat2Aon a VP ITC or nineteen 2 L injections of 200 M compound into 10 M Mat2A on an Auto iTC200. Data

8、areanalyzed and fit to a simple 1:1 binding model 1.MCE has not independently confirmed the accuracy of these methods. They are for reference only.Cell Assay 1 Huh-7 cells are seeded at a concentration of 15,000 cells per well for 6-h incubation with compound (PF-9366) and 4,000 cells per well for 7

9、2-h incubation with compound in 96-well plates in 200 L of growthmedium. NCI-H520 MAT2B knockdown cells are seeded at a concentration of 20,000 cells per well for 6 hincubation or 10,000 cells per well for 72 h incubation with compound in 96 well plates in 200 L of growthmedium. Cells are allowed to

10、 attach overnight at 37C with 5% CO2. A 5 solution of cycloleucine isprepared fresh from powder stock in growth medium. Other compounds (PF-9366) are diluted in 100%DMSO using a three-fold dilution scheme and further diluted in growth medium to give 0.5% DMSO final.Consistency of cellular confluence

11、 for each cell line is monitored with the IncuCyte Zoom live cell imager.Proliferation is measured using CellTiterGlo reagent. Growth media is removed from the cell plates followingcompound treatment and 80 L/well CellTiter Glo diluted 1:1 in PBS added. Luminescence is measured byan Plate Reader 1.M

12、CE 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 Mol Cell. 2019 Aug.See more customer validations on HYPERLINK / www.MedChemEREFERENCES2/3 Master of Small Molecules 您边的抑制剂师www.MedChemE1. Quinlan CL, et al. Targeting S-adenosylmethionine biosynthesis with a novel allosteric inhibitor of Mat2A. Nat Chem Biol. 2017Jul;13(7):785-792.McePdfHeightCauti