Alisertib

Manufactured by MedChemExpress

Sourced in United States

Alisertib is a small molecule inhibitor that targets the Aurora A kinase. It is a research reagent used in scientific studies to investigate the role of Aurora A kinase in various cellular processes.

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Close spelling variants of this product

Alisertib/MLN8237, (2 citations)

7 protocols using alisertib

Breast Cancer Cell Line Characterization

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Human breast cancer cell lines from the American Type Culture Collection were cultured in standard conditions. We confirmed negative mycoplasma testing and STR profile for each cell line. Alisertib was purchased from MedChem Express (Monmouth Junction, NJ). FRAX1036 was synthesized by AK and WW [22 (link)].

Korobeynikov V., Borakove M., Feng Y., Wuest W.M., Koval A.B., Nikonova A.S., Serebriiskii I., Chernoff J., Borges V.F., Golemis E.A, & Shagisultanova E. (2019). Combined inhibition of Aurora A and p21-activated kinase 1 as a new treatment strategy in breast cancer. Breast Cancer Research and Treatment, 177(2), 369-382.

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Evaluation of Selinexor, Temozolomide, and Alisertib

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The drug compounds used in this study were commercially obtained (i.e., selinexor from Selleckchem; temozolomide [TMZ] and alisertib from MedChemExpress) and resuspended as per the manufacturer's instructions for in vitro and in vivo use.

Nguyen R., Wang H., Sun M., Lee D.G., Peng J, & Thiele C.J. (2022). Combining selinexor with alisertib to target the p53 pathway in neuroblastoma. Neoplasia (New York, N.Y.), 26, 100776.

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Quantifying Viable Tumor Cells by MTT Assay

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MTT analysis was performed to quantify viable tumor cells in culture and to determine IC50 values after treatment with Adavivint, Alisertib, Berzosertib, Olaparib and Trametinib (all obtained from MedChemExpress, New York, USA). For dose response analyses, cells were seeded in a 96-well plate under standard culture conditions. After overnight culture, the cells were treated with a dilution series of the various inhibitors. 48 h after the initial treatment, all inhibitors were replenished and after 120 h MTT (5 mg/ml) was added to each well, followed by an incubation for 90 min at 37 °C. Subsequently, supernatants were aspirated, and precipitated formazan dye was solubilized with DMSO. Absorbance was measured at 540 nm using a Tecan infinite F50 spectrophotometer and viable tumor cells were calculated by the mean absorbance relative to DMSO control.

Viol F., Sipos B., Fahl M., Clauditz T.S., Amin T., Kriegs M., Nieser M., Izbicki J.R., Huber S., Lohse A.W, & Schrader J. (2022). Novel preclinical gastroenteropancreatic neuroendocrine neoplasia models demonstrate the feasibility of mutation-based targeted therapy. Cellular Oncology (Dordrecht), 45(6), 1401-1419.

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Angiogenic Inhibitors Modulate DIPG Tumor Growth

Cited 1 time

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Alisertib (MedChemExpress, Monmouth Junction, NJ), bortezomib (MedChemExpress), panobinostat (MedChemExpress), ponatinib (MedChemExpress), MSN1-Leu (synthesized in our lab, [20 (link)]) were sterilely prepared and dissolved in 0.5% DMSO and sterile PBS. Bevacizumab (SelleckChem) is water soluble, and was directly dissolved in sterile PBS. Drugs were administered by pipetting 15μL of drug solution directly on top of (but not contacting) the CAM tumor on embryonic development day 11, 13, 15. Each experiment contained 1–5 biological replicates per treatment group (at time of ultrasound) and there were at least three experimental replicates for each drug (n = 3–24 per drug treatment). Vehicle treatments (0.5% DMSO in sterile PBS) from all drug experimental replicates were combined to form one large vehicle group for further analysis (DIPGXIIIp* n = 38, DIPGIV n = 19).

Power E.A., Fernandez-Torres J., Zhang L., Yaun R., Lucien F, & Daniels D.J. (2022). Chorioallantoic membrane (CAM) assay to study treatment effects in diffuse intrinsic pontine glioma. PLoS ONE, 17(2), e0263822.

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Comparative Analysis of PKI with PLK4 Inhibition

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We studied 8 PKI with PLK4 cross-over potential: CFI-400945 (CAS#1338800-06-8, Cat#16850) (Cayman Chemical, Ann Arbor, MI, USA) [55 (link)]; CFI-400437 (CAS#1247000-76-5, Cat# SYN-1207) (SynKinase, San Diego, CA, USA) [60 (link)]; centrinone (CAS#1798871-30-3, Ludwig Institute for Cancer Research, New York, NY, USA) [59 (link)]; centrinone B (CAS#1798871-31-4, Ludwig Institute for Cancer Research, New York, NY, USA) [59 (link)]; KW-2449 (CAS# 1000669-72-6, Cat#HY-10339) (MedChemExpress, Monmouth Junction, NJ, USA) [63 (link)]; R1530 (CAS#882531-87-5, Cat#15225) (Cayman Chemical, Ann Arbor, MI, USA) [57 (link)]; axitinib (CAS#319460-85-0, Cat#13813) (Cayman Chemical, Ann Arbor, MI, USA) [61 (link)], and alisertib (CAS#1028486-01-2, Cat# HY-10971) (MedChemExpress, Monmouth Junction, NJ, USA) [97 (link)] (Figure 1).

Suri A., Bailey A.W., Tavares M.T., Gunosewoyo H., Dyer C.P., Grupenmacher A.T., Piper D.R., Horton R.A., Tomita T., Kozikowski A.P., Roy S.M, & Sredni S.T. (2019). Evaluation of Protein Kinase Inhibitors with PLK4 Cross-Over Potential in a Pre-Clinical Model of Cancer. International Journal of Molecular Sciences, 20(9), 2112.

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Cell Line Cultivation and Characterization

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Isogenic breast cancer cell lines, MCF7-WT/ RB-del were maintained in DMEM media containing 10% FBS. T47D cells were grown in RPMI media, supplemented with 10% FBS. CAMA-1 cells were purchased from ATCC and were cultured in EMEM media (Cat # 30–2003) that was supplemented with 10% FBS. All the cells lines were grown at 37 °C and 5% CO₂ and were confirmed to be mycoplasma free. Cell line authentication was performed using STR analysis. Palbociclib, Alisertib, and Birinapant were purchased from MedChemExpress (NJ, USA). Mk1775 was purchased from Chemitek (IN, USA). Fulvestrant, Everolimus, and Alpelisib were purchased from SelleckChem. MK1775 was purchased from ChemiTek. All the compounds were dissolved in DMSO to a final concentration of 10 mM. A customized drug library, comprising 311 compounds was purchased from SelleckChem.

Kumarasamy V., Nambiar R., Wang J., Rosenheck H., Witkiewicz A.K, & Knudsen E.S. (2022). RB loss determines selective resistance and novel vulnerabilities in ER-positive breast cancer models. Oncogene, 41(27), 3524-3538.

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Alisertib Inhibits Cancer Cell Viability

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Alisertib (name MLN8237, formula C₂₇H₂₀ClFN₄O₄, #HY-10971, Medchem Express, Sollentuna, Sweden), an orally active and selective Aurora A kinase inhibitor (IC50 = 1.2 nM), which binds to Aurora A kinase resulting in mitotic spindle abnormalities and mitotic accumulation, was dissolved in dimethyl sulfoxide (DMSO, Sigma, Darmstadt, Germany). The effect of Alisertib on CAL-120 and MDA-MB-231 cells viability was determined by a dose–response curve based on different drug concentrations of Alisertib ranging from 0.0001 µM to 20 µM, and DMSO as vehicle, for 72 h. Cell viability was measured using 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenylte-trazolium bromide (MTT) assay.

Skov N., Alves C.L., Ehmsen S, & Ditzel H.J. (2022). Aurora Kinase A and Bcl-xL Inhibition Suppresses Metastasis in Triple-Negative Breast Cancer. International Journal of Molecular Sciences, 23(17), 10053.

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