SBE 13 HCl

Catalog No.S7720 Batch:S772001

Technical Data

Formula	C₂₄H₂₈Cl₂N₂O₄
Molecular Weight	479.4	CAS No.	1052532-15-6
Solubility (25°C)*	In vitro	DMSO	95 mg/mL (198.16 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.)

Preparing Stock Solutions

Biological Activity

Description

SBE 13 HCl is a potent and selective PLK1 inhibitor with IC50 of 200 pM, >4000-fold selectivity over Aurora A kinase, Plk2 and Plk3.

Targets

PLK1 ^[1]

200 pM

In vitro

SBE 13 decreases cell proliferation in various cancer cell lines, and causes a G2/M arrest followed by apoptosis. ^[1] In primary cells, SBE 13 does not impair cell cycle and thus proliferation of primary cells. ^[2] SBE13 in combination with Enzastaurin displays a synergistic reduction of cell proliferation and enhanced apoptosis induction in HCT116(p53-/-) cells. ^[3]

Protocol (from reference)

Kinase Assay:^{^[1]}	Kinase assays To assay Plk1 and Aurora A kinase activity, cells are lysed after 13 hrs release in the presence of SBE13 after double thymidine block, and kinases are immunoprecipitated from lysates using antibodies as described. In brief, for each immunoprecipitation 800 µg of total protein were incubated with 1.5 µg Plk1 antibody cocktail, 3 µg Plk2 antibody, 3 µg Plk3 antibody, or 5 µg Aurora A antibody, respectively, for 2 hrs at 4°C on a rotator. Immunoprecipitated protein is collected using Protein G Agarose beads. The Plk1, Plk2 and Plk3 immunoprecipitates are incubated with 1 µg casein and with 1 µCi of [γ³²-P]ATP for 30 min at 37°C in kinase buffer. The Aurora A immunoprecipitates are incubated with 0.5 µl Histone and with 1 µCi of [γ³²-P]ATP for 60 min at room temperature in kinase buffer. Products from the kinase assays are fractionated on 10% Bis-Tris-polyacrylamide gels, and the phosphorylated substrate is visualized by autoradiography after an exposure of 12 to 36 hrs. An equal amount of immunoprecipitates is subjected to western blot analysis to confirm equal loading of Plk1, Plk2, Plk3 or Aurora A protein in kinase reactions. Immunoprecipitated Plk1 after 13 hrs release in the presence of SBE13 is assayed after de-phosphorylation using λ protein phosphatase and compared to kinase activity of endogenous immunoprecipitated Plk1. Activity of Plk1 kinase with and wiiiuithout de-phosphorylation is compared and the ratio between de-phosphorylated and “normal” endogenous immunoprecipitated Plk1 kinase activity is calculated.
Cell Assay:^sup>[1]	Cell lines HeLa, A431, HCT-15, HT-29, MCF-7, U2OS, LN-229, SKW 6.4, PC-3, Det-562, SK-BR-3, SK-OV-3 and A549nb cells Concentrations 100 μM Incubation Time 72 hours Method Cells are treated with SBE13 one day after subculturing. Control cells are incubated with normal culture medium. Concentrations of SBE13 ranged from 1 nM–100 µM. The growth rate of 1 x 10⁵ cells per 6-well is determined by counting cells at 24, 48 and 72 hours after treatment. Cell culture studies are performed in triplicate for each time point.

Kinase Assay:^{^[1]}

Kinase assays
To assay Plk1 and Aurora A kinase activity, cells are lysed after 13 hrs release in the presence of SBE13 after double thymidine block, and kinases are immunoprecipitated from lysates using antibodies as described. In brief, for each immunoprecipitation 800 µg of total protein were incubated with 1.5 µg Plk1 antibody cocktail, 3 µg Plk2 antibody, 3 µg Plk3 antibody, or 5 µg Aurora A antibody, respectively, for 2 hrs at 4°C on a rotator. Immunoprecipitated protein is collected using Protein G Agarose beads. The Plk1, Plk2 and Plk3 immunoprecipitates are incubated with 1 µg casein and with 1 µCi of [γ³²-P]ATP for 30 min at 37°C in kinase buffer. The Aurora A immunoprecipitates are incubated with 0.5 µl Histone and with 1 µCi of [γ³²-P]ATP for 60 min at room temperature in kinase buffer. Products from the kinase assays are fractionated on 10% Bis-Tris-polyacrylamide gels, and the phosphorylated substrate is visualized by autoradiography after an exposure of 12 to 36 hrs. An equal amount of immunoprecipitates is subjected to western blot analysis to confirm equal loading of Plk1, Plk2, Plk3 or Aurora A protein in kinase reactions. Immunoprecipitated Plk1 after 13 hrs release in the presence of SBE13 is assayed after de-phosphorylation using λ protein phosphatase and compared to kinase activity of endogenous immunoprecipitated Plk1. Activity of Plk1 kinase with and wiiiuithout de-phosphorylation is compared and the ratio between de-phosphorylated and “normal” endogenous immunoprecipitated Plk1 kinase activity is calculated.

Cell Assay:^sup>[1]

Cell lines
HeLa, A431, HCT-15, HT-29, MCF-7, U2OS, LN-229, SKW 6.4, PC-3, Det-562, SK-BR-3, SK-OV-3 and A549nb cells
Concentrations
100 μM
Incubation Time
72 hours
Method
Cells are treated with SBE13 one day after subculturing. Control cells are incubated with normal culture medium. Concentrations of SBE13 ranged from 1 nM–100 µM. The growth rate of 1 x 10⁵ cells per 6-well is determined by counting cells at 24, 48 and 72 hours after treatment. Cell culture studies are performed in triplicate for each time point.

References

Selleck's SBE 13 HCl has been cited by 7 publications

PLK1 inhibition dampens NLRP3 inflammasome-elicited response in inflammatory disease models [ J Clin Invest, 2023, 133(21)e162129]	PubMed: 37698938
PLK1 inhibition dampens NLRP3 inflammasome-elicited response in inflammatory disease models [ J Clin Invest, 2023, 133(21)e162129]	PubMed: 37698938
Inhibition of polo-like kinase 1 (PLK1) facilitates reactivation of gamma-herpesviruses and their elimination [ PLoS Pathog, 2021, 17(7):e1009764]	PubMed: 34297745
Effects of Aurora kinase A on mouse decidualization via Stat3-plk1-cdk1 pathway [ Reprod Biol Endocrinol, 2021, 19(1):162]	PubMed: 34715887
Inhibition of Polo-like kinase 1 (PLK1) facilitates the elimination of HIV-1 viral reservoirs in CD4 + T cells ex vivo [ Sci Adv, 2020, 6(29):eaba1941]	PubMed: 32832623
SF3B1 deficiency impairs human erythropoiesis via activation of p53 pathway: implications for understanding of ineffective erythropoiesis in MDS [ J Hematol Oncol, 2018, 11(1):19]	PubMed: 29433555
SF3B1 deficiency impairs human erythropoiesis via activation of p53 pathway: implications for understanding of ineffective erythropoiesis in MDS [ J Hematol Oncol, 2018, 11(1):19]	PubMed: 29433555

RETURN POLICY
Selleck Chemical’s Unconditional Return Policy ensures a smooth online shopping experience for our customers. If you are in any way unsatisfied with your purchase, you may return any item(s) within 7 days of receiving it. In the event of product quality issues, either protocol related or product related problems, you may return any item(s) within 365 days from the original purchase date. Please follow the instructions below when returning products.

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.

NOT FOR HUMAN, VETERINARY DIAGNOSTIC OR THERAPEUTIC USE.