TAK-632 - Raf 抑制劑 - 生命科學試劑 - MedChemExpress

1、Hotline: 400-820-3792Inhibitors Agonists Screening Librarieswww.MedChemETAK-632Cat. No.: HY-15767CAS No.: 1228591-30-7分式: CHFNOS分量: 554.52作靶點: Raf; Aurora Kinase作通路: MAPK/ERK Pathway; Cell Cycle/DNA Damage; Epigenetics儲存式: Powder -20C 3 years4C 2 yearsIn solvent -80C 6 months-20C 1 month溶解性數(shù)據(jù)體外實驗 DM

2、SO : 100 mg/mL (180.34 mM; Need ultrasonic)Mass Solvent1 mg 5 mg 10 mg Concentration制備儲備液1 mM 1.8034 mL 9.0168 mL 18.0336 mL5 mM 0.3607 mL 1.8034 mL 3.6067 mL10 mM 0.1803 mL 0.9017 mL 1.8034 mL請根據(jù)產(chǎn)品在不同溶劑中的溶解度，選擇合適的溶劑配制儲備液，并請注意儲備液的保存式和期限。體內(nèi)實驗 請根據(jù)您的實驗動物和給藥式選擇適當?shù)娜芙獍福渲魄罢埾扰渲瞥吻宓膬湟?，再依次添加助溶?為保證實驗結(jié)果的可靠性，體

3、內(nèi)實驗的作液，建議您現(xiàn)現(xiàn)配，當天使；澄清的儲備液可以根據(jù)儲存條件，適當保存；以下溶劑前的百分 指該溶劑在您配制終溶液中的體積占)：1. 請依序添加每種溶劑： 10% DMSO 40% PEG300 5% Tween-80 45% salineSolubility: 2.5 mg/mL (4.51 mM); Suspended solution; Need ultrasonic2. 請依序添加每種溶劑： 10% DMSO 90% corn oilSolubility: 2.5 mg/mL (4.51 mM); Clear solutionBIOLOGICAL ACTIVITY1/3 Master

4、 of Small Molecules 您邊的抑制劑師www.MedChemE物活性 TAK-632種有效的 pan-RAF 抑制劑，作于 CRAF，BRAFV600E 和 BRAFWT，IC50 分別為 1.4，2.4 和8.3 nM。IC50 & Target c-Raf Braf Aurora B PDGFR1.4 nM (IC50) 8.3 nM (IC50) 66 nM (IC50) 120 nM (IC50)PDGFR FGFR3 TIE2 IKK610 nM (IC50) 280 nM (IC50) 740 nM (IC50) 3700 nM (IC50)CDK1 CDK2 p3

5、8 GSK3790 nM (IC50) 580 nM (IC50) 600 nM (IC50) 500 nM (IC50)MEK13700 nM (IC50)體外研究 TAK-632 inhibits PDGFR, FGFR3, GSK3, CDK2, P38, PDGFR, TIE2, and CDK1 with a range of IC50values from 120-790 nM. CHK1, IKK, and MEK1 are inhibited over an IC50 range of 1400-1700 nM. With 1h of preincubation time, T

6、AK-632 inhibits BRAF and CRAF in an ATP competitive manner (at low ATPconcentrations BRAF IC50: 15 nM; CRAF: 8.1 nM). The respective biochemical activity of TAK-632 againstBRAF and CRAF reduces to IC50 values of 58 nM and 62 nM at high ATP concentrations.TAK-632demonstrates strong inhibition of pMEK

7、 and pERK in HMVII cells with IC50 values of 49 nM and 50 nM,respectively 1. TAK-632 shows strong antiproliferative effects both in A375 and SK-MEL-2 cells (GI50 of 40-190 nM in A375 cells and GI50 of 190-250 nM in SK-MEL-2 cells) 2.體內(nèi)研究 TAK-632 demonstrates dramatically improved solubility (740 g/m

8、L) in pH 6.8 phosphate buffer and exhibitssignificant oral absorption (at a dose of 25 mg/kg, AUC, 32.47 g h/mL; F, 51.7%) in rats. In a dog PK study,10 mg/kg administration of TAK-632 also shows superior oral bioavailability (F: 108%).Oral singleadministration of TAK-632 inhibits pERK in tumors at

9、8 h after its administration over a dose range of 1.9-24.1 mg/kg. In particular, 9.7-24.1 mg/kg dosing with TAK-632 strongly inhibits pERK levels to 11% of thecontrol. TAK-632 exhibits dose-dependent antitumor efficacy without severe body weight reduction over adose range of 3.9-24.1 mg/kg. Signific

10、ant tumor regression is observed at 9.7 mg/kg and 24.1 mg/kg(T/C=2.1% and 12.1%, respectively) 1. TAK-632 exhibits potent antitumor efficacy when orallyadministered at 60 mg/kg once daily (T/C=37%, P 2.PROTOCOLKinase Assay 2 Immunoprecipitated BRAF or CRAF is incubated with recombinant inactive MEK

11、(K97R) at 30C for 30minutes in kinase reaction buffer containing ATP/Mg2+. RAS/RAF wild-type (A431, CsFb, and HeLa), KRAS-mutant (A549, HCT-116, and MIA PaCa-2), and NRAS-mutant melanoma (GAK, HMV-II, and SK-MEL-2)cells are treated with TAK-632 (0, 0.32, 1.6, 8, 40, 200, 1000 and 5000 nM) at the ind

12、icated concentrationsfor 2 hours. Cell lysates are analyzed by Western blot analysis 2.MCE has not independently confirmed the accuracy of these methods. They are for reference only.Cell Assay 2 Cell viability is assessed (3 replicates) using the Sulforhodamine B assay or by the CellTiter-Glo lumine

13、scent2/3 Master of Small Molecules 您邊的抑制劑師www.MedChemEcell viability assay. The concentrations of TAK-632 that produced 50% growth inhibition (GI50) are calculatedusing PCP software. The combination index (CI) is calculated using CalcuSyn software. To investigate theantiproliferative activity of TAK

14、-632, we performed proliferation assays in various cell lines harboring mutatedBRAF, NRAS, or KRAS. HMV-II, SK-MEL-2, or A375 cells are cotreated with TAK-632 and TAK-733 at theindicated concentrations for 72 hours. Cell viability is measured. The CI value at EC50 is calculated. A375cells stably exp

15、ressing NRASQ61K or N-BRAF are cotreated with TAK-632 and TAK-733 at the indicatedconcentrations for 72 hours. Cell viability is measured. The CI value at EC50 is calculated 2.MCE has not independently confirmed the accuracy of these methods. They are for reference only.Animal Mice 2Administration 2

16、 The xenograft-implanted nude mice are used. Mice bearing SK-MEL-2 xenografts are treated once daily for21 consecutive days with vehicle or TAK-632 at the indicated concentrations (10 mice per each treatmentgroup). Day 0 indicates the beginning of treatment. Tumors are measured twice a week. Mice be

17、aring SK-MEL-2 xenografts are treated once daily (QD) for 3 days with vehicle, TAK-632 at 60 mg/kg (60 mpk), orTAK-632 at 120 mg/kg (120 mpk). Tumor xenografts are obtained at indicated time points after the finaltreatment and analyzed by Western blot analysis. Individual blots with dividing lines a

18、re combined from asingle electrophoresis gel. Bars represent densitometric analysis of phospho-ERK, normalized to vehicle-treated control (meanSD).MCE has not independently confirmed the accuracy of these methods. They are for reference only.戶使本產(chǎn)品發(fā)表的科研獻 Br J Pharmacol. 2019 Jun;176(12):2095-2108. Harvard Medical School LINCS LIBRARYSee more customer validations on HYPERLINK / www.Me