Lomeguatrib-PaTrin-2-DataSheet-生命科学试剂-MedChemExpressVIP

1、Hotline: 400-820-3792Inhibitors Agonists Screening Librarieswww.MedChemELomeguatribCat. No.: HY-13668CAS No.: 192441-08-0Synonyms: PaTrin-2分式: CHBrNOS分量: 326.17作靶点: DNA Methyltransferase作通路: 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 : 56 mg/mL (1

2、71.69 mM)* means soluble, but saturation unknown.Mass Solvent1 mg 5 mg 10 mg Concentration制备储备液1 mM 3.0659 mL 15.3294 mL 30.6589 mL5 mM 0.6132 mL 3.0659 mL 6.1318 mL10 mM 0.3066 mL 1.5329 mL 3.0659 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

3、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积占)：1. 请依序添加每种溶剂： 10% DMSO 40% PEG300 5% Tween-80 45% salineSolubility: 2.08 mg/mL (6.38 mM); Clear solution2. 请依序添加每种溶剂： 10% DMSO 90% (20% SBE-CD in saline)Solubility: 2.08 mg/mL (6.38 mM); Clear solution1/3 Master of Small Molecule

4、s 您边的抑制剂师www.MedChemE3. 请依序添加每种溶剂： 10% DMSO 90% corn oilSolubility: 2.08 mg/mL (6.38 mM); Clear solutionBIOLOGICAL ACTIVITY物活性 Lomeguatrib种有效的 MGMT 抑制剂，在细胞体系和 MCF-7 细胞体系中，IC50 值分别为 9 nM 和 -6nM。IC50 & Target MGMT MGMT6 nM (IC50, in MCF-7 cells) 9 nM (IC50)体外研究 Lomeguatrib (Compound 10) is a O6-methyl

5、guanine methyltransferase (MGMT) inhibitor, with an IC50 of 9nM in cell-free assay 1 and -6nM in MCF-7 cells. Lomeguatrib (10M) substantially increases the growthinhibitory effects of temozolomide in MCF-7 cells (D60=10M with Lomeguatrib vs 400M without) 2.体内研究 Lomeguatrib (20mg/kg i.p.) completely

6、inactivates MGMT within 2h, but shows no significant effect on tumorgrowth in MCF-7 xenografts 2.PROTOCOLKinase Assay 1 Briefly, 200 g of extracted cellular protein from HeLaS3 cells in 200 L of 70 mM HEPES buffer (with 1 mMdithiothreitol (DTT), 5 mM EDTA, pH 7.8) is incubated at 37C with a defined

7、concentration of Lomeguatrib(added as a DMSO solution). After 30 min an excess of 3H-methylated DNA (120000 cpm) is added, andthe incubation is continued for an additional 90 min. The reaction is stopped by the addition of 400 L TCA(13%), and the DNA is hydrolyzed by heating the reaction mixture for

8、 30 min at 98C. The precipitatedprotein is washed three times with 400-L portions of 5% TCA, solubilized in 0.1 N NaOH, and analyzed byliquid scintillation counting using the cocktail Rotiszint eco plus and a TRI-CARB. Enzyme activity isexpressed as fmol of 3Hmethyl transferred to TCA-insoluble prot

9、ein material per mg of total cellular protein.Percent inhibition is calculated relative to untreated control samples. Each assay is repeated three times, andIC50 values are determined graphically from plots of percent inhibition vs inhibitor concentration 1.MCE has not independently confirmed the ac

10、curacy of these methods. They are for reference only.Cell Assay 2 To determine toxicity, the MTT growth inhibition assay is employed. Cells (1000 per well) are plated into a96-well plate and following a 24h attachment period, Lomeguatrib is added to the cells. After 2h incubationwith Lomeguatrib (10

11、M) at 37C, 5% CO2, increasing doses of temozolomide or vehicle are added and thecells are incubated for a further 4-5 days. At the end of the exposure period, 150g MTT is added to eachwell and plates are incubated for 3h at 37C, 5% CO2. The media are removed and the formazan crystalsformed in the vi

12、able cells are solubilised in 200L DMSO. The absorbances at 540 and 690nm aredetermined using a ELISA plate reader and growth inhibition calculated as a percentage of the A540-A690 ofuntreated wells 2.MCE has not independently confirmed the accuracy of these methods. They are for reference only.2/3

13、Master of Small Molecules 您边的抑制剂师www.MedChemEAnimal Mice 2Administration 2 To assess the ability of Lomeguatrib to sensitise human breast tumour xenografts to the tumour growthinhibitory effects of temozolomide, groups of at least six nude mice are treated as follows: the vehicle controlgroup are gi

14、ven corn oil then 20% DMSO in PBS; the temozolomide only group are given corn oil thentemozolomide (100mg/kg/day); the Lomeguatrib only group are given Lomeguatrib (20mg/kg/day) thenDMSO in PBS, and the Lomeguatrib plus temozolomide group are given Lomeguatrib (20mg/kg/day) thentemozolomide (100mg/k

15、g/day). Drugs or vehicles are administered i.p. once daily for 5 days with aseparation of 1h. Up to 10 and at least six animals are assigned to each group, and mean tumour volume isstandardised across the groups at the start of the experiment: thus the control, Lomeguatrib, temozolomideand Lomeguatr

16、ib plus temozolomide groups had mean tumour volumes of 29.87.6 (range 19.0-38.7),33.214.7 (range 16.5-58.7), 35.110.9 (range 20.9-52.4) and 30.310.0 (range 20.7-44.5)mm3,respectively 2.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 Proc Nat

17、l Acad Sci U S A. 2019 Feb 19;116(8):2961-2966. Patent. US20180263995A1.See more customer validations on HYPERLINK / www.MedChemEREFERENCES1. Reinhard J, et al. Monosaccharide-linked inhibitors of O(6)-methylguanine-DNA methyltransferase (MGMT): synthesis, molecularmodeling, and structure-activity relationships. J Med Chem. 2001 Nov 22;44(24):4050-61.2. Clemons M, et al. O6-(4-bromothenyl)guanine reverses temozolomide resistance in human breast tumour MC