埃罗替尼作用机制- Medchemexpress- MCE中国

Product Data?SheetErlotinibCat. No.:CAS No.:分?式:分?量:作?靶点:作?通路:储存?式:HY-50896183321-74-6C??H??N?O?393.44EGFR; AutophagyJAK/STAT Signaling; Protein Tyrosine Kinase/RTK; AutophagyPowderIn solvent-20°C4°C-80°C-20°C3 years2 years6 months1 monthInhibitors?Agonists?Screening Libraries溶解性数据体外实验DMSO : ≥ 50 mg/mL (127.08 mM)H2O : < 0.1 mg/mL (insoluble)* \Mass1 mg5 mg10 mgSolventConcentration制备储备液1 mM5 mM10 mM2.5417 mL0.5083 mL0.2542 mL12.7084 mL2.5417 mL1.2708 mL25.4168 mL5.0834 mL2.5417 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；?旦配成溶液，请分装保存，避免反复冻融造成的产品失效。储备液的保存?式和期限：-80°C, 6 months; -20°C, 1 month。-80°C 储存时，请在 6 个?内使?，-20°C 储存时，请在 1 个?内使?。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。以下溶解?案都请先按照 In Vitro ?式配制澄清的储备液，再依次添加助溶剂： 为保证实验结果的可靠性，澄清的储备液可以根据储存条件，适当保存；体内实验的?作液，建议您现?现配，当天使?； 以下溶剂前显?的百分?是指该溶剂在您配制终溶液中的体积占?；如在配制过程中出现沉淀、析出现象，可以通过加热和/或超声的?式助溶1. 请依序添加每种溶剂：?0.5% CMC-Na/saline waterSolubility: 10 mg/mL (25.42 mM); Suspended solution; Need ultrasonic2. 请依序添加每种溶剂：?10% DMSO ?? 40% PEG300 ?? 5% Tween-80 ?? 45% salineSolubility: ≥ 2 mg/mL (5.08 mM); Clear solution此?案可获得 ≥ 2 mg/mL (5.08 mM，饱和度未知) 的澄清溶液。 以 1 mL ?作液为例，取 100 μL 20.0 mg/mL 的澄清 DMSO 储备液加到 400 μL PEG300 中，混合均匀；向上述体系中加? 50 μL Tween-80，混合均匀；然后继续加? 450 μL ?理盐?定容? 1 mL。Page 1 of 2 www.MedChemExpress.cn

3. 请依序添加每种溶剂：?10% DMSO ?? 90% (20% SBE-β-CD in saline)Solubility: 2.5 mg/mL (6.35 mM); Suspended solution; Need ultrasonic

此?案可获得 2.5 mg/mL (6.35 mM) 的均匀悬浊液，悬浊液可?于?服和腹腔注射。

以 1 mL ?作液为例，取 100 μL 25.0 mg/mL 的澄清 DMSO 储备液加到 900 μL 20% 的 SBE-β-CD ?理盐??溶液中，混合均匀。

4. 请依序添加每种溶剂：?10% DMSO ?? 90% corn oilSolubility: ≥ 2.5 mg/mL (6.35 mM); Clear solution

此?案可获得 ≥ 2.5 mg/mL (6.35 mM，饱和度未知) 的澄清溶液，此?案不适?于实验周期在半个?以上的实验。 以 1 mL ?作液为例，取 100 μL 25.0 mg/mL 的澄清 DMSO 储备液加到 900 μL ??油中，混合均匀。

5. 请依序添加每种溶剂：?50% PEG300 ?? 50% saline

Solubility: 10 mg/mL (25.42 mM); Suspended solution; Need ultrasonic

BIOLOGICAL ACTIVITY

?物活性

Erlotinib (CP-358774) 是?种直接作?的 EGFR 酪氨酸激酶抑制剂，对? EGFR 的 IC50 为 2 nM。Erlotinib 可降低完整肿瘤细胞的 EGFR ?磷酸化，IC50 为 20 nM。Erlotinib ?于治疗??细胞肺癌。

IC?? & Target

EGFR

2 nM (IC50, Cell Free Assay)

体外研究

Erlotinib (CP-358774) is also a potent inhibitor of the recombinant intracellular (kinase) domain of the EGFR, with an IC50 of 1 nM. The proliferation of DiFi cells is strongly inhibited by Erlotinib with an IC50 of 100 nM for an 8-day proliferation assay[1]. The combination of B-DIM and Erlotinib (2 μM) results in a significant inhibition of colony formation in BxPC-3 cells when compared with either agent alone. The combination of B-DIM and Erlotinib (2 μM) results in a significant induction of apoptosis only in BxPC-3 cells when compare with the apoptotic effect of either agent alone[2].

体内研究

Under the experimental conditions, the combination of B-DIM and Erlotinib (50 mg/kg, i.p.) treatment shows significant decrease (P <0.01) in tumor weight compared with untreated control[2]. Erlotinib (20 mg/kg, p.o.) significantly attenuates Cisplatin (CP)-induced body weight (BW) loss when compared to the CP+vehicle (V) rats (P<0.05). Erlotinib treatment significantly improves renal function in CP-N(normal control group, NC) rats. The CP+Erlotinib (E) rats show significant reduction of the levels of Serum creatinine (s-Cr) (P<0.05), blood urea nitrogen (BUN) (P<0.05), urinary N-acetyl-β-D-glucosaminidase (NAG) index (P<0.05), and significant increase of urine volume (UV) (P<0.05) and Cr clearance (Ccr) (P<0.05) compare to the CP+V rats[3]

PROTOCOL

Kinase Assay [1]

The kinase reaction is performed in 50 μL of 50 mM HEPES (pH 7.3), containing 125 mM NaCl, 24 mM MgCl2, 0.1 mM Na3VO4, 20 μM ATP, 1.6 μg/mL EGF, and 15 ng of EGFR, affinity purified from A431 cell membranes. The compound in DMSO is added to give a final DMSO concentration of 2.5%. Phosphorylation is initiated by addition of ATP and proceeded for 8 mm at room temperature, with constant shaking. The kinase reaction is terminated by aspiration of the reaction mixture and is washed 4 times with wash buffer. Phosphorylated PGT is measured by 25 mim of incubation with 50 μL per well HRP-conjugated PY54 antiphosphotyrosine antibody, diluted to 0.2 μg/mL in blocking buffer (3% BSA and 0.05% Tween 20 in PBS). Antibody is removed by aspiration, and the plate is washed 4 times with wash buffer. The colonmetric signal is developed by addition of TMB Microwell Peroxidase Substrate, 50 μL per well, and stopped by the addition of 0.09 M sulfuric acid, 50μL per well. Phosphotyrosine is estimated by measurement of

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absorbance at 450 nm. The signal for controls is typically 0.6-1.2 absorbance units, with essentially no back ground in wells without 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 viability of cells treated with B-DIM, Erlotinib, or the combination, BxPC-3 and MIAPaCa cells are plated (3,000-5,000 per well) in a 96-well plate and incubated overnight at 37°C. A range of concentrations for both B-DIM (10-50 μM) and Erlotinib (1-5 μM) is initially tested. Based on the initial results, the concentration of B-DIM (20 μM) and Erlotinib (2 μM) are chosen for all assays. The effects of B-DIM (20 μM), Erlotinib (2 μM), and the combination on BxPC-3 and MIAPaCa cells are determined by the standard MTT assay after 72 h and is repeated three times. The color intensity is measured by a Tecan microplate fluorometer at 595 nm. DMSO-treated cells are considered to be the untreated control and assigned a value of 100%. In addition to the above assay, we have also done clonogenic assay for assessing the effects of treatment[2].

MCE has not independently confirmed the accuracy of these methods. They are for reference only.

Animal

Administration [2][3]

Mice[2]

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 (50 mg/kg body weight), everyday i.p. for 15 days; and (d) B-DIM and Erlotinib, following schedule as for individual treatments. 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°C for future use and the other part is fixed in formalin and processed for paraffin block. H&E staining of fixed tissue section is used to confirm the presence of tumor(s) in each pancreas. Rats[3]

Six-week-old male Sprague-Dawley (SD) rats weighing 180 to 210 g are used. Cisplatin (CP) is freshly prepared in saline at a concentration of 1 mg/mL and then injected intraperitoneally in SD rats (n=28) at a dose 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+E, n=14) or vehicle (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 a normal control group (NC, n=5). The NC rats are given an equivalent volume of saline daily by oral gavage from day -1 to day 3. At day 4 (96 hours after CP injection), each rat is anesthetized and sacrificed by exsanguination after the cardiac puncture; blood is collected by cardiac puncture and kidneys are collected. Renal tissue is divided; separate portions are snap-frozen in liquid nitrogen or fixed in 2% paraformaldehyde/phosphate-buffered saline (PBS) for later use. All surgery is performed under diethyl ether gas anesthesia, and all efforts are made to minimize suffering.

MCE has not independently confirmed the accuracy of these methods. They are for reference only.

客户使?本产品发表的科研?献

????Sci Transl Med. 2024 Jul 18;10(450). pii: eaaq1093.????Nat Commun.?2024 Apr 18;10(1):1812????Oncogene. 2017 May 11;36(19):2643-2654.????J Invest Dermatol. 2024 Jan;139(1):224-234.????Mol Oncol. 2024 Mar;12(3):305-321.

See more customer validations on www.MedChemExpress.cnREFERENCES

[1].?Moyer JD, et al. Induction of apoptosis and cell cycle arrest by CP-358,774, an inhibitor of epidermal growth factor receptor tyrosine kinase. Cancer Res.

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1997, 57(21), 4838-4848.

[2].?Ali S, et al. Apoptosis-inducing effect of erlotinib is potentiated by 3,3'-diindolylmethane in vitro and in vivo using an orthotopic model of pancreatic cancer. Mol Cancer Ther, 2008, 7(6), 1708-1719.

[3].?Wada Y, et al. Epidermal growth factor receptor inhibition with erlotinib partially prevents cisplatin-induced nephrotoxicity in rats. PLoS One. 2014 Nov 12;9(11):e111728.

McePdfHeightCaution: Product has not been fully validated for medical applications. For research use only.

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