Pelitinib (EKB-569)

카탈로그 번호S1392 배치:S139202

인쇄

기술 자료

화학식

C24H23ClFN5O2
분자량 467.92 CAS 번호 257933-82-7
용해도 (25°C)* 시험관 내(In vitro) DMSO 13 mg/mL (27.78 mM)
Water Insoluble
Ethanol Insoluble
생체 내(In Vivo) (개별적으로 순서대로 용매를 제품에 첨가하십시오.)
Homogeneous suspension
CMC-NA
≥5mg/ml Taking the 1 mL working solution as an example, add 5 mg of this product to 1 ml of CMC-Na solution, mix evenly to obtain a homogeneous suspension with a final concentration of 5 mg/ml.
* <1 mg/ml은 약간 용해되거나 불용해됨을 의미합니다.
* Selleck은 모든 화합물의 용해도를 자체적으로 테스트하며, 실제 용해도는 게시된 값과 약간 다를 수 있습니다. 이는 정상적인 현상이며, 약간의 배치 간 변동으로 인해 발생합니다.
* 실온 배송 (안정성 테스트 결과 이 제품은 냉각 조치 없이 배송될 수 있음을 보여줍니다.)

원액 준비

생물학적 활성

설명 Pelitinib (EKB-569) is a potent irreversible EGFR inhibitor with IC50 of 38.5 nM. This compound also slightly inhibits Src, MEK/ERK and ErbB2 with IC50s of 282 nM, 800 nM and 1255 nM, respectively. Phase2.
표적
EGFR Src MEK/ERK ErbB2 Raf
38.5 nM 282 nM 800 nM 1.255 μM 3.353 μM
시험관 내(In vitro) Pelitinib (EKB-569) displays much higher inhibitory activity against EGFR, compared with the closely related c-erbB-2, as well as other kinases such as Src, Cdk4, c-Met, Raf, and MEK/ERK, with IC50 ranging from 282 nM for Src to >20 μM for Cdk4. Consistently, this compound treatment significantly inhibits the autophosphorylation of EGFR but not c-Met in A431 cells. It potently inhibits the proliferation of normal human keratinocytes (NHEK), as well as A431 and MDA-468 tumor cells with IC50 of 61 nM, 125 nM, and 260 nM, respectively, while displaying little activity against MCF-7 cells with IC50 of 3.6 μM. This chemical inhibits EGF-induced phosphorylation of EGFR in A431 and NHEK cells with IC50 of 20-80 nM, as well as the phosphorylation of STAT3 with IC50 of 30-70 nM. It at 75-500 nM also specifically inhibits the activation of AKT and ERK1/2, without affecting NF-κB pathway. In NHEK cells, this compound also potently inhibits TGF-α mediated EGFR activation with IC50 of 56 nM, as well as activation of STAT3 and ERK1/2 with IC50 of 60 nM and 62 nM, respectively.
생체 내(In Vivo) A single oral dose of 10 mg/kg Pelitinib (EKB-569) potently inhibits the EGFR phosphorylation in A431 xenografts with over-expressed EGFR, by 90% within 1 hour, and by >50% after 24 hours. Administration of this compound at 20 mg/kg/day inhibits tumorigenesis in APCMin/+ mice by 87%, equivalent to the effect of used with 2 times doses of EKI-785 (40 mg/kg/day), consistent with greater in vivo potency. This chemical selectively inhibits EGFR signaling in airway epithelial cells in vivo. In the mouse model of airway epithelial remodeling that is inducible by viral infection and features a delayed but permanent switch to goblet cell metaplasia, this compound treatment at 20 mg/kg/day corrects all 3 aspects of epithelial remodeling, by completely blocking the increase of ciliated cells and decrease of Clara cells, and significantly inhibiting the metaplasia of goblet cells.
특징 An improved version of EKI-785.

프로토콜 (참조)

키나아제 분석:

[1]
  • Autophosphorylation of EGFR in cells

    For experiments using cells in culture, A431 cells are treated with various concentrations of Pelitinib (EKB-569) for 2.75 hours before co-incubation with 100 ng/mL EGF for 0.25 hour. Cells are washed twice with cold phosphate-buffered saline (PBS) before adding to lysis buffer (10 mM Tris, pH 7.5, 5 mM ethylenediamine tetra-acetic acid (EDTA), 150 mM NaCl, 1% Triton X-100, 1% Sodium deoxycholate, 0.1 % SDS, 1 mM PMSF, 10 mg/mL pepstatin A, 10 mg/mL leupeptin, 20 KIU/mL aprotinin, 2 mM sodium orthovanadate, and 100 mM sodium fluoride) for 20 minutes on ice, before immunoprecipitation and SDS-PAGE-immunoblotting. For immunoprecipitation, cultured cells are placed in cold lysis buffer and immediately homogenized on ice with a polytron with several pulses. The homogenate is first centrifuged at 2500 rpm (20 minutes, 4 °C) and then again at 14,000 rpm in a microcentrifuge (10 minutes, 4 °C). Supernatants (1000 μg protein) are incubated for 2 hours at 4 °C with 15 mL of EGFR polyclonal antibody. After 2 hours, 50 μL of protein G plus/protein A agarose beads is added and incubated with constant rotation for 2 hours at 4 °C. After washing with lysis buffer, beads are boiled for 2 minutes in Laemmli sample buffer. Proteins are then resolved by SDS-PAGE, transferred to immobilon membrane and probed overnight with an anti-phosphotyrosine antibody conjugated with horseradish peroxidase (HRP). Membranes are developed using the ECL reagent. Total EGFR protein is determined by stripping membranes and re-probing with receptor-specific antibodies. Quantitation of bands is done by densitometry, using ImageQuant software with a Molecular Dynamics laser transmittance scanner.
세포 분석:

[2]
  • 세포주

    NHEK, A431, MCF-7, and MDA-468

  • 농도

    Dissolved in DMSO, final concentrations ~10 μM

  • 배양 시간

    5 days

  • 방법

    Cells are seeded in 96-well dishes, and after 2 hours, Pelitinib (EKB-569) is added and incubated for 5 days. After incubation, the medium is removed from each well and fresh medium (150 μL) + 1 mg/mL MTT solution (50 μL) is added. After incubation for 2 hours at 37 °C, the medium is replaced with 150 μL DMSO, and absorbance at 540 nm in each well is determined. The IC50 is calculated by linear regression of the data.
동물 연구:

[1]
  • 동물 모델

    Athymic nu/nu female mice bearing subcutaneous A431 tumors, or APCMin/+ male mice, a murine model of human familial adenomatous polyposis (FAP)

  • 용량

    10, or 20 mg/kg/day

  • 투여

    Oral gavage

참조

  • https://pubmed.ncbi.nlm.nih.gov/10973323/
  • https://pubmed.ncbi.nlm.nih.gov/14749472/
  • https://pubmed.ncbi.nlm.nih.gov/16453019/

고객 제품 검증

HBMEC were preincubated with the indicated concentrations of either ErB1/2/4 inhibitor (Pelitinib; EKB-569), and cells were then infected for 4 h with the unencapsulated strain, MC58 siaD. The numbers of adherent (black bars) and invasive (gray bars) bacteria were determined in a gentamicin protection assay. The graphs show the percentages of adhesion and invasion of inhibitor-treated cells, relative to control cells (means 盨D of three independent experiments performed in duplicate). *, P < 0.05.

데이터 출처 [ Infect Immun , 2014 , 82(3), 1243-55 ]

<p>IGF-1 and IL-1β mediated induction of Bcl-2 expression involves EGFR.   (A)  Bcl-2 mRNA levels in AALEBs treated with IGF-1 or IL-1β in the presence or absence of two EGFR tyrosine kinase inhibitors, EKB-569 (1 μM) or PD153035 (1 μM). </p>

데이터 출처 [ J Immunol , 2012 , 188, 4581-4589 ]

Immunofluorescence Assay. PDCD4, GRA3, and overlapping are represented in red, green, and yellow, respectively. Group I TKIs sunitinib and AZD9291 show similar results to that of the negative control with PDCD4 and GRA3 well localized inside the nuclei. Mild disruption of PDCD4 in the nuclei is observed with Group II TKIs gefitinib, erlotinib, and AG1478. Group III TKIs neratinib, afatinib, and pelitinib show comparable changes to that of pyrimethamine 5 μM, with complete disruption of PDCD4 and GRA3, without any localization. The assays were repeated 3 times, and each experiment was performed in triplicate. A representative result is shown (x1,000).

데이터 출처 [ , , Korean J Parasitol, 2017, 55(5):491-503 ]

Selleck's Pelitinib (EKB-569) 인용됨 11 출판물

Integrative analysis of drug response and clinical outcome in acute myeloid leukemia [ Cancer Cell, 2022, S1535-6108(22)00312-9] PubMed: 35868306
EGFR Inhibition Potentiates FGFR Inhibitor Therapy and Overcomes Resistance in FGFR2 Fusion-Positive Cholangiocarcinoma [ Cancer Discov, 2022, 12(5):1378-1395] PubMed: 35420673
The multi-kinase inhibitor afatinib serves as a novel candidate for the treatment of human uveal melanoma [ Cell Oncol (Dordr), 2022, 45(4):601-619] PubMed: 35781872
Biomarker LEPRE1 induces pelitinib-specific drug responsiveness by regulating ABCG2 expression and tumor transition states in human leukemia and lung cancer [ Sci Rep, 2022, 12(1):2928] PubMed: 35190588
A Genome-Scale CRISPR Screen Identifies the ERBB and mTOR Signalling Networks as Key Determinants of Response to PI3K Inhibition in Pancreatic Cancer [ Mol Cancer Ther, 2020, 5;molcanther.1131.2019] PubMed: 32371585
Mapping phospho-catalytic dependencies of therapy-resistant tumours reveals actionable vulnerabilities. [ Nat Cell Biol, 2019, 21(6):778-790] PubMed: 31160710
Suppressors for Human Epidermal Growth Factor Receptor 2/4 (HER2/4): A New Family of Anti-Toxoplasmic Agents in ARPE-19 Cells [Kim YH Korean J Parasitol, 2017, 55(5):491-503] PubMed: 29103264
Sensitivity of Melanoma Cells to EGFR and FGFR Activation but Not Inhibition is Influenced by Oncogenic BRAF and NRAS Mutations. [Garay T, et al. Pathol Oncol Res, 2015, 10.1007/s12253-015-9916-9] PubMed: 25749811
Selective inhibition of human solute carrier transporters by multikinase inhibitors [Johnston RA, et al. Drug Metab Dispos, 2014, 42(11):1851-7] PubMed: 25165131
Role of epidermal growth factor receptor signaling in the interaction of Neisseria meningitidis with endothelial cells [Slanina H et al. Infect Immun, 2014, 82(3):1243-55] PubMed: 24379285

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