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CHIR-124 Chk inhibitor

Cat.No.S2683

CHIR-124 is a novel and potent Chk1 inhibitor with IC50 of 0.3 nM in a cell-free assay. It shows 2,000-fold selectivity against Chk2, 500- to 5,000-fold less activity against CDK2/4 and Cdc2.
CHIR-124 Chk inhibitor Chemical Structure

Chemical Structure

Molecular Weight: 419.91

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Quality Control

Cell Culture, Treatment & Working Concentration

Cell Lines Assay Type Concentration Incubation Time Formulation Activity Description PMID
human MDA-MB-435 cells Cytotoxic assay Cytotoxicity against human MDA-MB-435 cells, EC50=0.08 μM
human MDA-MB-435 cells Cytotoxic assay Cytotoxicity against human MDA-MB-435 cells in presence of camptothecin
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Chemical Information, Storage & Stability

Molecular Weight 419.91 Formula

C23H22ClN5O

 Storage (From the date of receipt)
CAS No. 405168-58-3 Download SDF Storage of Stock Solutions

Synonyms N/A Smiles C1CN2CCC1C(C2)NC3=C(C(=O)NC4=C3C=C(C=C4)Cl)C5=NC6=CC=CC=C6N5

Solubility

In vitro
Batch:

DMSO : Insoluble
(Moisture-contaminated DMSO may reduce solubility. Use fresh, anhydrous DMSO.)

Water : Insoluble

Ethanol : Insoluble

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Mass Concentration Volume Molecular Weight

In vivo
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Mechanism of Action

Targets/IC50/Ki
Chk1 [1]
(Cell-free assay)
0.3 nM
FLT3 [1]
(Cell-free assay)
5.8 nM
PDGFR [1]
(Cell-free assay)
6.6 nM
GSK-3 [1]
(Cell-free assay)
23.3 nM
In vitro

CHIR-124 is a quinolone-based small molecule that is structurally unrelated to other known inhibitors of Chk1. This compound interacts synergistically with topoisomerase poisons (e.g., Camptothecin or SN-38) in causing growth inhibition in a variety of cancer cell lines, including breast carcinoma (MDA-MB-231 and MDA-MB-435) and colon carcinoma (SW-620 and Colo205), all of which contains the mutant p53 gene. It abrogates the SN-38-induced S and G2-M checkpoints and potentiates apoptosis in MDA-MD-435 breast cancer cells. The abrogation of the G2-Mcheckpoint and induction of apoptosis by this chemical are enhanced by the loss of p53. [1] This compound also potently targets other kinases such as PDGFR and Flt3 with IC50 of 6.6 nM and 5.8 nM, respectively. [2]
Kinase Assay
Chk1 Assay
For the Chk1 assay, the kinase domain is expressed in Sf9 insect cells, and a biotinylated cdc25c peptide containing the consensus Chk1/Chk2 phosphorylation site (*)(biotin-[AHX]SGSGS*GLYRSPSMP-ENLNRPR[CONH2]) is used as the substrate. A dilution series of CHIR-124 is mixed with a kinasereaction buffer containing a final concentration of 30 mM Tris-HCl(pH 7.5), 10 mM MgCl2, 2 mM DTT, 4 mM EDTA, 25 mMβ-glycerophosphate, 5 mM MnCl2, 0.01% bovine serum albumin, 1.35 nM CHK1 kinase domain, 0.5 μM peptide substrate, and 1 AM unlabeled ATP, plus 5 nM 33Pγ-labeled ATP (specific activity = 2,000 Ci/mmol). Reactions and detection of the phosphate transfer are carried out by a radioactive method. Reactions are incubated at room temperature for 1 to 4 hours and the phosphorylated peptide captured on streptavidin-coated microtiter plates containing stop reaction buffer (25 mM EDTA [ethylenediaminetetraacetic acid], 50 mMHEPES, pH 7.5). Phosphorylated peptide is measured with the DELFIA TRF system using a Europium-labeled anti-phosphotyrosine antibody PT66. The concentration of this compound for IC50 is calculated using nonlinear regression with XL-Fit data analysis software.
In vivo

CHIR-124 potentiates the growth inhibitory effects by abrogating the G2-M checkpoint and increasing tumor apoptosis in an orthotopic breast cancer xenograft model.
References

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