1、Product Data SheetDacomitinibCat. No.: HY-13272CAS No.: 1110813-31-4分式: CHClFNO分量: 469.94作靶點: EGFR; Apoptosis作通路: JAK/STAT Signaling; Protein Tyrosine Kinase/RTK; Apoptosis儲存式: Powder -20C 3 years4C 2 yearsIn solvent -80C 6 months-20C 1 month溶解性數據體外實驗 DMSO : 41.67 mg/mL (88.67 mM; Need ultrasonic)H2

2、O : 0.1 mg/mL (insoluble)SolventMass1 mg 5 mg 10 mgConcentration制備儲備液1 mM 2.1279 mL 10.6397 mL 21.2793 mL5 mM 0.4256 mL 2.1279 mL 4.2559 mL10 mM 0.2128 mL 1.0640 mL 2.1279 mL請根據產品在不同溶劑中的溶解度選擇合適的溶劑配制儲備液；旦配成溶液，請分裝保存，避免反復凍融造成的產品失效。儲備液的保存式和期限：-80C, 6 months; -20C, 1 month。-80C 儲存時，請在 6 個內使，-20C 儲存時，請在 1

3、 個內使。體內實驗請根據您的實驗動物和給藥式選擇適當的溶解案。以下溶解案都請先按照 In Vitro 式配制澄清的儲備液，再依次添加助溶劑：為保證實驗結果的可靠性，澄 的儲備液可以根據儲存條件，適當保存；體內實驗的作液，建議您現現配，當天使； 以下溶劑前顯的百分 指該溶劑在您配制終溶液中的體積占；如在配制過程中出現沉淀、析出現象，可以通過加熱和/或超聲的式助溶1. 請依序添加每種溶劑： 10% DMSO 40% PEG300 5% Tween-80 45% salineSolubility: 2.5 mg/mL (5.32 mM); Clear solution此案可獲得 2.5 mg/mL

4、(5.32 mM，飽和度未知) 的澄清溶液。以 1 mL 作液為例，取 100 L 25.0 mg/mL 的澄 DMSO 儲備液加到 400 L PEG300 中，混合均勻；向上述體系中加50 L Tween-80，混合均勻；然后繼續(xù)加 450 L 理鹽定容 1 mL。2. 請依序添加每種溶劑： 10% DMSO 90% (20% SBE-CD in saline)Solubility: 2.5 mg/mL (5.32 mM); Suspended solution; Need ultrasonicPage 1 of 2 www.MedChemE此案可獲得 2.5 mg/mL (5.32 mM

5、) 的均勻懸濁液，懸濁液可于服和腹腔注射。以 1 mL 作液為例，取 100 L 25.0 mg/mL 的澄 DMSO 儲備液加到 900 L 20% 的 SBE-CD 理鹽溶液中，混合均勻。3. 請依序添加每種溶劑： 10% DMSO 90% corn oilSolubility: 2.5 mg/mL (5.32 mM); Clear solution此案可獲得 2.5 mg/mL (5.32 mM，飽和度未知) 的澄 溶液，此案不適于實驗周 期在半個以上的實驗。以 1 mL 作液為例，取 100 L 25.0 mg/mL 的澄 DMSO 儲備液加到 900 L 油中，混合均勻。BIOLOG

6、ICAL ACTIVITY物活性 Dacomitinib種特異，不可逆的 ERBB 家族抑制劑，作于 EGFR，ERBB2 和 ERBB4 的 IC50 分別為 6 nM，45.7 nM和 73.7 nM。IC & Target EGFR ErbB2 ErbB46 nM (IC50) 45.7 nM (IC50) 73.7 nM (IC50)體外研究 Dacomitinib (PF00299804) effectively inhibits the in vitro kinase activity of wild-type EGFR (IC50=6 nM)with similarefficac

7、y. Dacomitinib also effectively inhibits wild-type ERBB2 with IC50 of 45.7 nM. In H441, an IC50 is reached withDacomitinib but only at a very high concentration (4 M) and likely reflects off-target effects. In cell lines wild-type forboth EGFR and K-ras (H322, H1819, and Calu-3), Gefitinib and Dacom

8、itinib both effectively inhibit growth of H1819and Calu-3 cells but not of H322 cells. Dacomitinib is a pan-ERBB inhibitor and most EGFR mutant cell lines expressmultiple ERBB family members, the effects on EGFR phosphorylation could potentially be indirect. Dacomitinibinhibits EGFR phosphorylation

9、in all of the different EGFR T790M proteins whereas Gefitinib is ineffective even at 10 M. In the NIH3T3 cells, phosphorylation of EGFR L858R/T790M is completely inhibited by 1 nM Dacomitinib, whereas100 nM or greater is required to inhibit EGFR WT/T790M or Del/T790M1. The HER2-amplified cell lines

10、are mostsensitive to growth inhibition by Dacomitinib (IC501 M in 14 of 16 lines; 87.5%) as compared with 5 of 28 (17.9%)of HER2-nonamplified lines (excluding immortalized lines)2.體內研究 To evaluate the efficacy of Dacomitinib, xenografts in nu/nu mice are generated using HCC827 GFP and HCC827Del/T790

11、M cells and treated the mice with Dacomitinib. Dacomitinib (10 mg/kg/d by daily oral gavage) effectivelyinhibits the growth of HCC827 GFP xenografts. In contrast, HCC827 Del/T790M xenografts are resistant to Gefitinib,whereas Dacomitinib treatment is substantially more effective at inhibiting growth

12、 of this xenograft model1.PROTOCOLKinase Assay 1 The catalytic domains of ERBB1, ERBB2, and ERBB4 tagged with glutathione S-transferase are expressed in insect cellsand purified. ELISA-based enzyme assays and IC50 determinations for ERBB1, ERBB2, and ERBB4 are performed.Enzyme assays and IC50 determ

13、inations for all other kinases used in this study are performed1.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. Briefl

14、y, day after plating, Dacomitinib is added at 10 M and 2-fold dilutions over 12 concentrations arecarried out to generate a dose-response curve. Control wells without the drug are also seeded. The cells are countedon day 1 when the drug is added, as well as after 6 days when the experiment ended. Af

15、ter the trypsinization cells areplaced in an Isotone solution and immediately counted using a Coulter Z1 particle counter. The suspension culturesare counted using a Coulter Vi-Cell counter2.MCE has not independently confirmed the accuracy of these methods. They are for reference only.Page 2 of 3 ww

16、w.MedChemEAnimal Mice1Administration 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 lung cancer cells (in 0.2 mL ofPBS) are inoculated s.c. into the lo

17、wer-right quadrant of the flank of each mouse. Five mice are inoculated with eitherHCC827-GFP or HCC827-Del/T790M cells in the Gefitinib treatment group. Tumors are measured twice weekly usingcalipers, and volume is calculated using the following formula: lengthwidth20.52. Mice are monitored daily f

18、orbody weight and general condition. Mice are randomized to treatment when the mean tumor volume is 400 to 500mm3. Gefitinib is administered at 150 mg/kg/d by daily oral gavage. Dacomitinib is administered at 10 mg/kg/d bydaily oral gavage. The experiment is terminated when the mean size of the cont

19、rol tumors reached 2000 mm3.MCE has not independently confirmed the accuracy of these methods. They are for reference only.戶使本產品發(fā)表的科研獻 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. Harvard Medical School LINCS LIBRARYSee more customer validations on HYPERLINK www.MedChemE www.MedChemEREFERENCES1. Engelman JA, et al