AT7519 HCl

카탈로그 번호S7808 배치:S780801

인쇄

기술 자료

화학식

C16H18Cl3N5O2
분자량 418.71 CAS 번호 902135-91-5
용해도 (25°C)* 시험관 내(In vitro) DMSO 52 mg/mL (124.19 mM)
Water 43 mg/mL (102.69 mM)
Ethanol 28 mg/mL (66.87 mM)
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.
Clear solution
Saline

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 30.000mg/ml (71.65mM) Taking the 1 mL working solution as an example, add 30 mg of this product to 1 ml of physiological saline (0.9% NaCL solution), mix evenly to make it clear, The mixed solution should be used immediately for optimal results. 
* <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 AT7519 HCl is a multi-CDK inhibitor for CDK1, 2, 4, 6 and 9 with IC50 of 10-210 nM in cell-free assays. It is less potent to CDK3 and little active to CDK7. Phase 2.
표적
CDK9/CyclinT
(Cell-free assay)		 CDK5/p35
(Cell-free assay)		 CDK2/CyclinA
(Cell-free assay)		 GSK-3β
(Cell-free assay)		 CDK4/CyclinD1
(Cell-free assay)		 더 보기
<10 nM 13 nM 47 nM 89 nM 100 nM
시험관 내(In vitro) AT7519 is an ATP competitive CDK inhibitor with a Ki value of 38 nM for CDK1. AT7519 is inactive against all non-CDK kinases with the exception of GSK3β (IC50 = 89 nM). AT7519 shows potent antiproliferative activity in a variety of human tumor cell lines with IC50 values ranging from 40 nM for MCF-7 to 940 nM for SW620 consistent with the inhibition of CDK1 and CDK2. AT7519 induces dose-dependent cytotoxicity in multiple myeloma (MM) cell lines with IC50 values ranging from 0.5 to 2 μM at 48 hours, with the most sensitive cell lines being MM.1S (0.5 μM) and U266 (0.5 μM) and the most resistant MM.1R (>2 μM). It does not induce cytotoxicity in peripheral blood mononuclear cells (PBMNC). AT7519 partially overcomes the proliferative advantage conferred by IL6 and IGF-1 as well as the protective effect of bone marrow stromal cells (BMSCs). AT7519 induces rapid dephosphorylation of RNA pol II CTD at serine 2 and serine 5 sites, and leads to the inhibition of transcription, partially contributing to AT7519 induced cytotoxicity of MM cells. AT7519 induces activation of GSK-3β by down-regulating GSK-3β phosphorylation, which also contributes to AT7519 induced apoptosis independent of the inhibition of transcription.
생체 내(In Vivo) A twice daily dosing of AT7519 (9.1 mg/kg) causes tumor regression of both early-stage and advanced-stage s.c. tumors in the HCT116 and HT29 colon cancer xenograft models. AT7519 treatment (15 mg/kg) inhibits tumor growth and prolongs the median overall survival of mice in the human MM xenograft mouse model in association with increased caspase 3 activation.

프로토콜 (참조)

키나아제 분석:[1]
  • In vitro Kinase Assays

    Kinase assays for CDK1, CDK2 and GSK3-β are all carried out in a radiometric filter binding format. Assays for CDK5 are in DELFIA format and for CDKs 4 and 6 in ELISA format. For CDKs 1 and 2, the relevant CDK and 0.12 μg/mL Histone H1 are incubated in 20 mM MOPS, pH 7.2, 25 mM β-glycerophosphate, 5 mM EDTA, 15 mM MgCl2, 1 mM sodium orthovanadate, 1 mM DTT, 0.1 mg/mL BSA, 45 μM ATP (0.78 Ci/mmol) and different concentrations of AT7519 for 2 or 4 hours respectively. For GSK3-β, the relevant enzyme and 5 μM glycogen synthase peptide 2 along with 10 mM MOPS pH 7.0, 0.1 mg/mL BSA, 0.001% Brij-35, 0.5% glycerol, 0.2 mM EDTA, 10 mM MgCl2, 0.01% β-mercaptoethanol, 15 μM ATP (2.31 Ci/mmol) and different concentrations of AT7519 are incubated for 3 hours. Assay reactions are stopped by adding an excess of orthophosphoric acid and filtered using Millipore MAPH filter plates. The plates are then washed, scintillant added and radioactivity measured by scintillation counting on a Packard TopCount. For CDK5, CDK5/p35 and 1μM of a biotinylated Histone H1 peptide (Biotin-PKTPKKAKKL) are incubated in 25 mM Tris-HCl, pH 7.5, 2.5 mM MgCl2, 0.025% Brij-35, 0.1 mg/mL BSA, 1 mM DTT, 15 μM ATP and different concentrations of AT7519 for 30 minutes. Assay reactions are stopped using EDTA, transferred to Neutravidin-coated plates and phosphorylated peptide quantified by means of a rabbit phospho-cdk1 substrate polyclonal antibody and DELFIA europium-labelled anti-rabbit IgG secondary antibody using time-resolved fluorescence at λex=335nm, λem=620nm. For CDK 4 and 6 assays, plates are coated with GST- pRb769-921 and blocked with Superblock. CDK4 or 6 is incubated with 15 mM MgCl2, 50 mM HEPES, pH 7.4, 1 mM DTT, 1 mM EGTA, pH 8.0, 0.02% Triton X-100, 2.5% DMSO and different concentrations of AT7519; the reaction is initiated by addition of ATP. After 30 minutes, reactions are stopped by the addition of 0.5 M EDTA pH 8.0. Plates are then washed and incubated for one hour with the primary antibody (anti- p-Rb Serine 780) diluted in Superblock followed by secondary antibody (alkaline phosphatase linked anti-rabbit) for a further hour. Plates are developed using the Attophos system and fluorescence read on a Spectramax Gemini plate reader at excitation 450 nm and emission 580 nm. In all cases, IC50 values are calculated from replicate curves, using GraphPad Prism software.
세포 분석:[2]
  • 세포주

    MM.1S, MM.1R, RPMI8226, U266, RPMI8266, RPMI-Dox40, OPM1 cells, primary MM cells and PBMNCs

  • 농도

    Dissolved in DMSO at a concentration of 10 mM, final concentrations 0.25-4 μM

  • 배양 시간

    24 or 48 hours

  • 방법

    Cells are incubated with different concentrations of AT7519 for 24 or 48 hours at 37°C. Cell viability is assessed by measuring 3-(4,5-dimethylthiazol-2-yl)-2,5 diphenyl tetrasodium bromide (MTT) dye absorbance. DNA synthesis is measured by tritiated thymidine uptake (3H-TdR). Apoptosis is assessed by using Annexin V/PI staining. The percentage of cells undergoing apoptosis is defined as the sum of early apoptosis (Annexin V-positive cells) and late apoptosis (Annexin V-positive and PI-positive cells
동물 연구:[2]
  • 동물 모델

    Male SCID mice inoculated subcutaneously with MM.1S cells

  • 용량

    15 mg/kg/day

  • 투여

    Dosed i.p.

참조

  • https://pubmed.ncbi.nlm.nih.gov/19174555/
  • https://pubmed.ncbi.nlm.nih.gov/20101221/

고객 제품 검증

(c) Effect of AT7519 treatment on RNA pol II occupancy at the PRCC gene. MM1.S cells were treated with either DMSO vehicle (blue) or 2 μM AT7519 (brown) for 6 h, followed by RNA pol II ChIP-seq analysis. Twenty-fold magnifications of the rpm/bp scale of these gene tracks are shown in the right panel to show the difference in reads for elongating RNA pol II. TR, RNA pol II traveling ratio.(d) Genome-wide binding average RNA pol II (ChIP-seq) on active promoters and gene bodies following treatment of MM1.S cells with DMSO vehicle (blue) or 2 μM of AT7519 (brown) for 6 h. Magnification of the rpm/bp scale at gene bodies is shown in the inset. The inset includes RNA polymerase II traveling ratio distributions (TR, mean) derived from MM1.S cells treated with DMSO (blue) or 2 μM AT7519 (red).(e) Chemical structures of the pan-CDK inhibitor AT7519 and its biotinylated counterpart bio-AT7519.(f) In vitro kinase assays with recombinant cyclin T-CDK9 complex in the presence of increasing concentrations of AT7519 or bio-AT7519. The derived IC50 values for each compound are shown.(g) Effect of AT7519 and bio-AT7519 on MM1.S cell proliferation. Cells were treated with varying concentrations of drug for 72 h as indicated. The derived EC50 values for each compound are shown.

데이터 출처 [ , , Nat Biotechnol, 2014, 32(1): 92-96. ]

(C) Western blot analyses following treatment with the indicated doses of cyclin-dependent kinase (CDK) inhibitors for apoptosis markers, poly(ADP-ribose) polymerase 1 (PARP-1), caspase 3, cleaved form of caspase 3 and β-actin.

데이터 출처 [ , , Int J Oncol, 2018, 53(2):703-712 ]

Selleck's AT7519 HCl 인용됨 16 출판물

Interleukin-15 enhanced the survival of human γδT cells by regulating the expression of Mcl-1 in neuroblastoma [ Cell Death Discov, 2022, 8(1):139] PubMed: 35351861
O-GlcNAc transferase maintains metabolic homeostasis in response to CDK9 inhibition [ Glycobiology, 2022, cwac038] PubMed: 35708495
Cdk2 suppresses IL-23 expression and the onset of severe acute pancreatitis [ Immun Inflamm Dis, 2022, 10(6):e631] PubMed: 35634959
High-content image-based analysis and proteomic profiling identifies Tau phosphorylation inhibitors in a human iPSC-derived glutamatergic neuronal model of tauopathy [ Sci Rep, 2021, 11(1):17029] PubMed: 34426604
The cyclin-dependent kinase inhibitor AT7519 augments cisplatin's efficacy in ovarian cancer via multiple oncogenic signaling pathways [ Fundam Clin Pharmacol, 2021, 10.1111/fcp.12709] PubMed: 34212421
Development of a miRNA-controlled dual-sensing system and its application for targeting miR-21 signaling in tumorigenesis [ Exp Mol Med, 2020, 10.1038/s12276-020-00537-z] PubMed: 33311703
Inhibition of O-GlcNAc Transferase Renders Prostate Cancer Cells Dependent on CDK9 [ Molecular Cancer Research, 2020, 1512-1521] PubMed: 32611550
Inhibition of Cyclin-dependent Kinase (CDK) Decreased Survival of NB4 Leukemic Cells: Proposing a p53-Independent Sensitivity of Leukemic Cells to Multi-CDKs Inhibitor AT7519 [ Iran J Pharm Res, 2020, 19(3):144-155] PubMed: 33680018
Fibroblast growth factor receptor influences primary cilium length through an interaction with intestinal cell kinase [ Proc Natl Acad Sci U S A, 2019, 116(10):4316-4325] PubMed: 30782830
CDK Blockade Using AT7519 Suppresses Acute Myeloid Leukemia Cell Survival through the Inhibition of Autophagy and Intensifies the Anti-leukemic Effect of Arsenic Trioxide [ Iran J Pharm Res, 2019, 18(Suppl1):119-131] PubMed: 32802093

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