BMS-265246

Catalog No.S2014 Batch:S201402

Technical Data

Formula	C₁₈H₁₇F₂N₃O₂
Molecular Weight	345.34	CAS No.	582315-72-8
Solubility (25°C)*	In vitro	DMSO	35 mg/mL (101.34 mM)
		Water	Insoluble
		Ethanol	Insoluble
* <1 mg/ml means slightly soluble or insoluble. * Please note that Selleck tests the solubility of all compounds in-house, and the actual solubility may differ slightly from published values. This is normal and is due to slight batch-to-batch variations. * Room temperature shipping (Stability testing shows this product can be shipped without any cooling measures.)

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Biological Activity

Description

BMS-265246 is a potent and selective CDK1/2 inhibitor with IC50 of 6 nM/9 nM in a cell-free assay. It is 25-fold more selective for CDK1/2 than CDK4.

Targets

CDK1/CyclinB ^[1] (Cell-free assay)	CDK2/CyclinE ^[1] (Cell-free assay)	CDK4/CyclinD ^[1] (Cell-free assay)
6 nM	9 nM	230 nM

In vitro

BMS265246 inhibits the activity of CDK4/cycD with IC50 of 0.23 μM and prevents A2780 Cytox with IC50 of 0.76 μM. BMS265246 which binds to CDK2 shows the inhibitor resides coincident with the ATP purine binding site and forms important H-bonds with Leu83 on the protein backbone. BMS265246 exhibits CDK1 and CDK2 potency that is 25- and 11-fold more potent versus CDK1 and CDK2, respectively. BMS265246 represents the most potent CDK/CDK2 selective analogue from this chemotype. ^[1] A recent study shows that BMS-265246 inhibits cell proliferation with EC50 ranging from 0.293 μM-0.492 μM in HCT-116 cells. After treatment of BMS-265246, the dominant cell populations are G2-arrested cells having 4N DNA content, large round nuclei, and low DNA intensity. ^[2]

Protocol (from reference)

Kinase Assay:^{^[1]}	CDK1/Cyclin B1 Kinase Assay Kinase reactions consists of 100 ng of baculovirus expressed GST-CDK1/cyclin B1 complex, 1 μg histone H1, 0.2 μCi 33P γ-ATP, 25 μM ATP in 50 μL of kinase buffer (50 mM Tris, pH 8.0, 10 mM MgCl₂, 1 mM EGTA, 0.5 mM DTT). Reactions are incubated for 45 min at 30 °C and stopped by the addition of cold trichloroacetic acid (TCA) to a final concentration of 15%. TCA precipitates are collected onto GF/C unifilter plates using a Filtermate universal harvester, and the filters are quantitated using a TopCount 96 well liquid scintillation counter. Dose response curves are generated to determine the concentration required to inhibit 50% of kinase activity (IC50). BMS265246 is dissolved at 10 mM in DMSO and evaluated at six concentrations, each in triplicate. The final concentration of DMSO in the assay equals 2%. IC50 values are derived by nonlinear regression analysis and have a coefficient of variance (SD/mean, n = 6) = 16%.
Cell Assay:^{^[2]}	Cell lines HCT116 Concentrations 0-5 μM Incubation Time 24 hours Method HCT-116 cells are plated onto 96-well dishes. For each well, the cell density is calculated by counting the number of objects (cells) per field of view, and averaging across all fields for a given well. For a treatment compound, cell density is converted to a percentage relative to the plate-averaged cell density from DMSO treatment (i.e., 100% corresponds to the average cell density for DMSO treatment). Logistic regression curve fits are done using TIBCO Spotfire, and the concentration at which the curve crosses 50% is reported as the EC50 of BMS-265246.

Kinase Assay:^{^[1]}

CDK1/Cyclin B1 Kinase Assay
Kinase reactions consists of 100 ng of baculovirus expressed GST-CDK1/cyclin B1 complex, 1 μg histone H1, 0.2 μCi 33P γ-ATP, 25 μM ATP in 50 μL of kinase buffer (50 mM Tris, pH 8.0, 10 mM MgCl₂, 1 mM EGTA, 0.5 mM DTT). Reactions are incubated for 45 min at 30 °C and stopped by the addition of cold trichloroacetic acid (TCA) to a final concentration of 15%. TCA precipitates are collected onto GF/C unifilter plates using a Filtermate universal harvester, and the filters are quantitated using a TopCount 96 well liquid scintillation counter. Dose response curves are generated to determine the concentration required to inhibit 50% of kinase activity (IC50). BMS265246 is dissolved at 10 mM in DMSO and evaluated at six concentrations, each in triplicate. The final concentration of DMSO in the assay equals 2%. IC50 values are derived by nonlinear regression analysis and have a coefficient of variance (SD/mean, n = 6) = 16%.

Cell Assay:^{^[2]}

Cell lines
HCT116
Concentrations
0-5 μM
Incubation Time
24 hours
Method
HCT-116 cells are plated onto 96-well dishes. For each well, the cell density is calculated by counting the number of objects (cells) per field of view, and averaging across all fields for a given well. For a treatment compound, cell density is converted to a percentage relative to the plate-averaged cell density from DMSO treatment (i.e., 100% corresponds to the average cell density for DMSO treatment). Logistic regression curve fits are done using TIBCO Spotfire, and the concentration at which the curve crosses 50% is reported as the EC50 of BMS-265246.

References

Selleck's BMS-265246 has been cited by 12 publications

CDC7 inhibition induces replication stress-mediated aneuploid cells with an inflammatory phenotype sensitizing tumors to immune checkpoint blockade [ Nat Commun, 2023, 10.1038/s41467-023-43274-3]	PubMed: 37980406
Nascent transcriptome reveals orchestration of zygotic genome activation in early embryogenesis [ Curr Biol, 2022, 32(19):4314-4324.e7]	PubMed: 36007528
Non-catalytic allostery in α-TAT1 by a phospho-switch drives dynamic microtubule acetylation [ J Cell Biol, 2022, 221-11e202202100]	PubMed: 36222836
Olaparib Induced Senescence is Bypassed through G2/M Checkpoint Override in Olaparib Resistant Prostate Cancer [ Mol Cancer Ther, 2022, molcanther.0604.2021]	PubMed: 35086956
Bromodomain-Containing Protein BRD4 Is Hyperphosphorylated in Mitosis [ Cancers (Basel), 2020, 12(6):E1637]	PubMed: 32575711
Inducing and exploiting vulnerabilities for the treatment of liver cancer [ Nature, 2019, 10.1038/s41586-019-1607-3]	PubMed: 31578521
Inducing and exploiting vulnerabilities for the treatment of liver cancer. [ Nature, 2019, 574(7777):268-272]	PubMed: 31578521
The TRAPP complex mediates secretion arrest induced by stress granule assembly. [ EMBO J, 2019, 38(19):e101704]	PubMed: 31429971
CDK9 modulates circadian clock by attenuating REV-ERBα activity. [ Biochem Biophys Res Commun, 2019, 513(4):967-973]	PubMed: 31005255
Nuclear Nestin deficiency drives tumor senescence via lamin A/C-dependent nuclear deformation. [ Nat Commun, 2018, 9(1):3613]	PubMed: 30190500

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SHIPPING AND STORAGE
Selleck products are transported at room temperature. If you receive the product at room temperature, please rest assured, the Selleck Quality Inspection Department has conducted experiments to verify that the normal temperature placement of one month will not affect the biological activity of powder products. After collecting, please store the product according to the requirements described in the datasheet. Most Selleck products are stable under the recommended conditions.

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