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CHIR-124 Chk inhibitor

Name: CHIR-124
Brand: Selleck Chemicals
SKU: S2683
Rating: 5 (47 reviews)

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.

Chemical Structure

Molecular Weight: 419.91

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Quality Control

Batch: Purity: 99.48%

99.48

Related Targets	CDK HSP PD-1/PD-L1 ROCK Wee1 DNA/RNA Synthesis Microtubule Associated Ras KRas Aurora Kinase
Other Chk Inhibitors	Prexasertib (LY2606368) Dihydrochloride CCT245737 (SRA737) AZD7762 Rabusertib (LY2603618) MK-8776 (SCH 900776) PF-477736 Prexasertib (LY2606368) BML-277 (Chk2 Inhibitor II) GDC-0575 SAR-020106

Cell Culture, Treatment & Working Concentration

Cell Lines	Assay Type	Activity Description
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
Click to View More Cell Line Experimental Data

Chemical Information, Storage & Stability

Molecular Weight	419.91	Formula	C₂₃H₂₂ClN₅O	Storage (From the date of receipt)	3 years-20°Cpowder
CAS No.	405168-58-3	Download SDF		Storage of Stock Solutions	1 year-80°Cin solvent 1 month-20°Cin solvent
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

Molarity Calculator

Mass	Concentration	Volume	Molecular Weight
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Dilution Calculator Molecular Weight Calculator

In vivo Batch:
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.

In vivo Formulation Calculator (Clear solution)

Step 1: Enter information below (Recommended: An additional animal making an allowance for loss during the experiment)

Dosage: mg/kg Average weight of animals: g Dosing volume per animal:μL Number of animals:

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% DMSO + % + % Tween 80 + % ddH₂O

%DMSO+ %

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Working concentration: mg/ml;

Method for preparing DMSO master liquid: mg drug pre-dissolved in μL DMSO ( Master liquid concentration mg/mL, Please contact us first if the concentration exceeds the DMSO solubility of the batch of drug. )

Method for preparing in vivo formulation: Take μL DMSO master liquid, next addμL PEG300, mix and clarify, next addμL Tween 80, mix and clarify, next add μL ddH₂O, mix and clarify.

Method for preparing in vivo formulation: Take μL DMSO master liquid, next add μL Corn oil, mix and clarify.

Note: 1. Please make sure the liquid is clear before adding the next solvent.
2. Be sure to add the solvent(s) in order. You must ensure that the solution obtained, in the previous addition, is a clear solution before proceeding to add the next solvent. Physical methods such as vortex, ultrasound or hot water bath can be used to aid dissolving.

Mechanism of Action

Targets/IC₅₀/Ki	Chk1 (Cell-free assay) 0.3 nM FLT3 (Cell-free assay) 5.8 nM PDGFR (Cell-free assay) 6.6 nM GSK-3 (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 Cell Cycle Checkpoints and potentiates apoptosis in MDA-MD-435 breast cancer cells. The abrogation of the G2-M Cell Cycle Checkpoint and induction of apoptosis by this chemical are enhanced by the loss of p53. This compound also potently targets other kinases such as PDGFR and Flt3 with IC50 of 6.6 nM and 5.8 nM, respectively.
Kinase Assay	Chk1 Assay
Kinase 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]SGSGSGLYRSPSMP-ENLNRPR[CONH2]) is used as the substrate. A dilution series of CHIR-124 is mixed with a kinase reaction buffer containing a final concentration of 30 mM Tris-HCl (pH 7.5), 10 mM MgCl₂, 2 mM DTT, 4 mM EDTA, 25 mM β-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 mM HEPES, 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 Cell Cycle Checkpoint and increasing tumour apoptosis in an orthotopic breast cancer xenograft model.
References	[1] https://pubmed.ncbi.nlm.nih.gov/17255282/ [2] https://pubmed.ncbi.nlm.nih.gov/20068082/

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