Erlotinib-CP-358774-DataSheet-生命科學(xué)試劑-MedChemExpress

1、Hotline: 400-820-3792Inhibitors Agonists Screening Librarieswww.MedChemEErlotinibCat. No.: HY-50896CAS No.: 183321-74-6Synonyms: CP-358774; NSC 718781; OSI-774分式: CHNO分量: 393.44作靶點(diǎn): EGFR; Autophagy作通路: JAK/STAT Signaling; Protein Tyrosine Kinase/RTK; Autophagy儲(chǔ)存式: Powder -20C 3 years4C 2 yearsIn sol

2、vent -80C 6 months-20C 1 month溶解性數(shù)據(jù)體外實(shí)驗(yàn) DMSO : 50 mg/mL (127.08 mM)* means soluble, but saturation unknown.Mass Solvent1 mg 5 mg 10 mg Concentration制備儲(chǔ)備液1 mM 2.5417 mL 12.7084 mL 25.4168 mL5 mM 0.5083 mL 2.5417 mL 5.0834 mL10 mM 0.2542 mL 1.2708 mL 2.5417 mL請(qǐng)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(chǔ)備液，并請(qǐng)注意儲(chǔ)備液的保存式和期

3、限。體內(nèi)實(shí)驗(yàn) 請(qǐng)根據(jù)您的實(shí)驗(yàn)動(dòng)物和給藥式選擇適當(dāng)?shù)娜芙獍?，配制前?qǐng)先配制澄清的儲(chǔ)備液，再依次添加助溶劑(為保證實(shí)驗(yàn)結(jié)果的可靠性，體內(nèi)實(shí)驗(yàn)的作液，建議您現(xiàn)現(xiàn)配，當(dāng)天使；澄清的儲(chǔ)備液可以根據(jù)儲(chǔ)存條件，適當(dāng)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(qǐng)依序添加每種溶劑： 10% DMSO 40% PEG300 5% Tween-80 45% salineSolubility: 2 mg/mL (5.08 mM); Clear solution2. 請(qǐng)依序添加每種溶劑： 10% DMSO 90% corn oilSolubility: 2 mg/mL (5.08 mM); Cle

4、ar solution1/4 Master of Small Molecules 您邊的抑制劑師www.MedChemEBIOLOGICAL ACTIVITY物活性 Erlotinib種治療細(xì)胞肺癌的藥物。抑制純化 EGFR 激酶的 IC50 為2 nM。IC50 & Target EGFR2 nM (IC50, Cell Free Assay)體外研究 Erlotinib (CP-358,774) is also a potent inhibitor of the recombinant intracellular (kinase) domain of the EGFR,with an IC

5、50 of 1 nM. The proliferation of DiFi cells is strongly inhibited by Erlotinib with an IC50 of 100 nMfor an 8-day proliferation assay 1. The combination of B-DIM and Erlotinib (2 M) results in a significantinhibition of colony formation in BxPC-3 cells when compared with either agent alone. The comb

6、ination of B-DIM and Erlotinib (2 M) results in a significant induction of apoptosis only in BxPC-3 cells when comparewith the apoptotic effect of either agent alone 2.體內(nèi)研究 Under the experimental conditions, the combination of B-DIM and Erlotinib (50 mg/kg, i.p.) treatment showssignificant decrease

7、(P 2. Erlotinib (20 mg/kg, p.o.) significantly attenuates Cisplatin (CP)-induced bodyweight (BW) loss when compared to the CP+vehicle (V) rats (P 3PROTOCOLKinase Assay 1 The kinase reaction is performed in 50 L of 50 mM HEPES (pH 7.3), containing 125 mM NaCl, 24 mMMgCl2, 0.1 mM Na3VO4, 20 M ATP, 1.6

8、 g/mL EGF, and 15 ng of EGFR, affinity purified from A431 cellmembranes. The compound in DMSO is added to give a final DMSO concentration of 2.5%. Phosphorylationis initiated by addition of ATP and proceeded for 8 mm at room temperature, with constant shaking. Thekinase reaction is terminated by asp

9、iration of the reaction mixture and is washed 4 times with wash buffer.Phosphorylated PGT is measured by 25 mim of incubation with 50 L per well HRP-conjugated PY54antiphosphotyrosine antibody, diluted to 0.2 g/mL in blocking buffer (3% BSA and 0.05% Tween 20 in PBS).Antibody is removed by aspiratio

10、n, and the plate is washed 4 times with wash buffer. The colonmetric signalis developed by addition of TMB Microwell Peroxidase Substrate, 50 L per well, and stopped by the additionof 0.09 M sulfuric acid, 50L per well. Phosphotyrosine is estimated by measurement of absorbance at 450nm. The signal f

11、or controls is typically 0.6-1.2 absorbance units, with essentially no back ground in wellswithout AlP, EGFR, or POT and is proportional to the time of incubation for 10 mm 1.MCE has not independently confirmed the accuracy of these methods. They are for reference only.Cell Assay 2 To test the viabi

12、lity of cells treated with B-DIM, Erlotinib, or the combination, BxPC-3 and MIAPaCa cells areplated (3,000-5,000 per well) in a 96-well plate and incubated overnight at 37C. A range of concentrationsfor both B-DIM (10-50 M) and Erlotinib (1-5 M) is initially tested. Based on the initial results, the

13、concentration of B-DIM (20 M) and Erlotinib (2 M) are chosen for all assays. The effects of B-DIM (20 M),Erlotinib (2 M), and the combination on BxPC-3 and MIAPaCa cells are determined by the standard MTTassay after 72 h and is repeated three times. The color intensity is measured by a Tecan micropl

14、atefluorometer at 595 nm. DMSO-treated cells are considered to be the untreated control and assigned a valueof 100%. In addition to the above assay, we have also done clonogenic assay for assessing the effects of2/4 Master of Small Molecules 您邊的抑制劑師www.MedChemEtreatment 2.MCE has not independently c

15、onfirmed the accuracy of these methods. They are for reference only.Animal Mice 2Administration 23 Female ICR-SCID (6-7 weeks old) mice are randomized into the following treatment groups (n=7): (a)untreated control; (b) only B-DIM (50 mg/kg body weight), intragastric once every day; (c) Erlotinib (5

16、0 mg/kgbody weight), everyday i.p. for 15 days; and (d) B-DIM and Erlotinib, following schedule as for individualtreatments. All mice are killed on day 3 following last dose of treatment, and their body weight is determined.One part of the tissue is rapidly frozen in liquid nitrogen and stored at 70

17、C for future use and the other partis fixed in formalin and processed for paraffin block. H&E staining of fixed tissue section is used to confirmthe presence of tumor(s) in each pancreas.Rats 3Six-week-old male Sprague-Dawley (SD) rats weighing 180 to 210 g are used. Cisplatin (CP) is freshlyprepare

18、d in saline at a concentration of 1 mg/mL and then injected intraperitoneally in SD rats (n=28) at adose of 7 mg/kg on day 0. To investigate the effect of Erlotinib, 28 CP-N rats are divided into two groups.Separate groups (n=14) each of animals are administered with either Erlotinib (20 mg/kg) (CP+

19、E, n=14) orvehicle (CP+V, n=14) daily by oral gavage from day -1 (24 hours prior to the CP injection) to day 3. Vehicle-treated groups receive an equivalent volume of saline. Five male SD rats at the age of 6 weeks are used as anormal control group (NC, n=5). The NC rats are given an equivalent volu

20、me of saline daily by oral gavagefrom day -1 to day 3. At day 4 (96 hours after CP injection), each rat is anesthetized and sacrificed byexsanguination after the cardiac puncture; blood is collected by cardiac puncture and kidneys are collected.Renal tissue is divided; separate portions are snap-fro

21、zen in liquid nitrogen or fixed in 2% paraformaldehyde/phosphate-buffered saline (PBS) for later use. All surgery is performed under diethyl ethergas anesthesia, and all efforts are made to minimize suffering.MCE has not independently confirmed the accuracy of these methods. They are for reference o

22、nly.戶使本產(chǎn)品發(fā)表的科研獻(xiàn) Sci Transl Med. 2018 Jul 18;10(450). pii: eaaq1093. Nat Commun. 2019 Apr 18;10(1):1812 Oncogene. 2017 May 11;36(19):2643-2654. J Invest Dermatol. 2019 Jan;139(1):224-234. Mol Oncol. 2018 Mar;12(3):305-321.See more customer validations on HYPERLINK / www.MedChemEREFERENCES1. Moyer JD, et al. Induction of apoptosis and cell cycle arrest by CP-358,77