Dacomitinib-PF-00299804-DataSheet-MedChemExpress

1、Hotline: 400-820-3792Inhibitors Agonists Screening Librarieswww.MedChemEDacomitinibCat. No.: HY-13272CAS No.: 1110813-31-4Synonyms: PF-00299804; PF-299804分式: CHClFNO分量: 469.94作靶点: EGFR作通路: JAK/STAT Signaling; Protein Tyrosine Kinase/RTK储存式: Powder -20C 3 years4C 2 yearsIn solvent -80C 6 months-20C 1

2、 month溶解性数据体外实验 DMSO : 50 mg/mL (106.40 mM; Need ultrasonic)H2O : 40% PEG300 5% Tween-80 45% salineSolubility: 2.5 mg/mL (5.32 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 (5.32 mM); Suspended solution; Need ultrasonic1/3 Master of Small Molecules 您边的抑制剂师

3、www.MedChemE3. 请依序添加每种溶剂： 10% DMSO 90% corn oilSolubility: 2.5 mg/mL (5.32 mM); Clear solutionBIOLOGICAL ACTIVITY物活性 Dacomitinib种特异，不可逆的 ERBB 家族抑制剂，作于 EGFR，ERBB2 和 ERBB4 的 IC50 分别为 6nM，45.7 nM 和 73.7 nM。IC50 & Target EGFR ErbB2 ErbB46 nM (IC50) 45.7 nM (IC50) 73.7 nM (IC50)体外研究 Dacomitinib (PF002998

4、04) effectively inhibits the in vitro kinase activity of wild-type EGFR (IC50=6 nM)withsimilar efficacy. Dacomitinib also effectively inhibits wild-type ERBB2 with IC50 of 45.7 nM. In H441, an IC50is reached with Dacomitinib but only at a very high concentration (4 M) and likely reflects off-target

5、effects.In cell lines wild-type for both EGFR and K-ras (H322, H1819, and Calu-3), Gefitinib and Dacomitinib botheffectively inhibit growth of H1819 and Calu-3 cells but not of H322 cells. Dacomitinib is a pan-ERBB inhibitorand most EGFR mutant cell lines express multiple ERBB family members, the ef

6、fects on EGFRphosphorylation could potentially be indirect. Dacomitinib inhibits EGFR phosphorylation in all of the differentEGFR T790M proteins whereas Gefitinib is ineffective even at 10 M. In the NIH3T3 cells, phosphorylation ofEGFR L858R/T790M is completely inhibited by 1 nM Dacomitinib, whereas

7、 100 nM or greater is required toinhibit EGFR WT/T790M or Del/T790M 1. The HER2-amplified cell lines are most sensitive to growthinhibition by Dacomitinib (IC50 2.体内研究 To evaluate the efficacy of Dacomitinib, xenografts in nu/nu mice are generated using HCC827 GFP andHCC827 Del/T790M cells and treat

8、ed the mice with Dacomitinib. Dacomitinib (10 mg/kg/d by daily oralgavage) effectively inhibits the growth of HCC827 GFP xenografts. In contrast, HCC827 Del/T790Mxenografts are resistant to Gefitinib, whereas Dacomitinib treatment is substantially more effective atinhibiting growth of this xenograft

9、 model 1.PROTOCOLKinase Assay 1 The catalytic domains of ERBB1, ERBB2, and ERBB4 tagged with glutathione S-transferase are expressedin insect cells and purified. ELISA-based enzyme assays and IC50 determinations for ERBB1, ERBB2, andERBB4 are performed. Enzyme assays and IC50 determinations for all

10、other kinases used in this study areperformed 1.MCE has not independently confirmed the accuracy of these methods. They are for reference only.Cell Assay 2 Cells are seeded in duplicate at 5103 to 5104 cells per well in 24-well plates, and growth inhibition data iscalculated. Briefly, day after plat

11、ing, Dacomitinib is added at 10 M and 2-fold dilutions over 12concentrations are carried out to generate a dose-response curve. Control wells without the drug are alsoseeded. The cells are counted on day 1 when the drug is added, as well as after 6 days when the experimentended. After the trypsiniza

12、tion cells are placed in an Isotone solution and immediately counted using a2/3 Master of Small Molecules 您边的抑制剂师www.MedChemECoulter Z1 particle counter. The suspension cultures are counted using a Coulter Vi-Cell counter 2.MCE has not independently confirmed the accuracy of these methods. They are

13、for reference only.Animal Mice 1Administration 1 Nude mice (nu/nu; 6-8 weeks old) are used for in vivo studies. Mice are anesthetized using a 2% isoflurane(Baxter) inhalation oxygen mixture. A suspension of 5106 HCC827-GFP or HCC827-Del/T790M lungcancer cells (in 0.2 mL of PBS) are inoculated s.c. i

14、nto the lower-right quadrant of the flank of each mouse.Five mice are inoculated with either HCC827-GFP or HCC827-Del/T790M cells in the Gefitinib treatmentgroup. Tumors are measured twice weekly using calipers, and volume is calculated using the followingformula: lengthwidth20.52. Mice are monitore

15、d daily for body weight and general condition. Mice arerandomized to treatment when the mean tumor volume is 400 to 500 mm3. Gefitinib is administered at 150mg/kg/d by daily oral gavage. Dacomitinib is administered at 10 mg/kg/d by daily oral gavage. Theexperiment is terminated when the mean size of

16、 the control tumors reached 2000 mm3.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 Sci Transl Med. 2018 Jul 18;10(450). pii: eaaq1093. Mol Cancer Ther. 2018 Mar;17(3):603-613. RSC Adv., 2018, 8, 38733 PLoS One. 2019 Apr 4;14(4):e0214598. H

17、arvard Medical School LINCS LIBRARYSee more customer validations on HYPERLINK / www.MedChemEREFERENCES1. Engelman JA, et al. PF00299804, an irreversible pan-ERBB inhibitor, is effective in lung cancer models with EGFR and ERBB2mutations that are resistant to gefitinib. Cancer Res. 2007 Dec 15;67(24):11924-32.2. Kalous O, et al. Dacomitinib (PF-00299804), an irreversible Pan-HER inhibitor, inhibits proliferation of HER2-amplified breast cancer celllines resistant to tr