Alisertib (MLN8237)

Alisertib (MLN 8237) is a selective Aurora A inhibitor with an IC50 of 1.2 nM.

Signaling Pathways >> Cell Cycle/DNA Damage >> Aurora Kinase

Signaling Pathways >> Epigenetics >> Aurora Kinase

Signaling Pathways >> Autophagy >> Autophagy

Research Areas >> Cancer

Aurora A:1.2 nM (IC50)

Alisertib leads the MM cells to mitotic spindle abnormalities, mitotic accumulation, as well as inhibition of cell proliferation through apoptosis and senescence. Alisertib up-regulates p53 and tumor suppressor genes p21 and p27[1]. The decreased activity of MLN8054/Alisertib for the T217D/W277E Aurora A/TPX2 complex may reflect the increased affinity for ATP induced by cofactor binding to Aurora A[2]. Alisertib inhibits cell proliferation with IC50 values ranging from 15 to 469 nM in different tumer cell lines[3].

Alisertib (Alisertib, 30 mg/kg, p.o.) significantly reduces tumor burden and increases overall survival in xenograft-murine model of human-MM[1]. Alisertib (20, 30 mg/kg, p.o.) causes tumor growth inhibition in solid tumor xenograft models and regressions in in vivo models of lymphoma, and reduces FLT uptake in HCT-116 xenograft tumors[3].

To measure Aurora A activity, 25 ng (12.5 mM final concentration) or 250 ng (125 nM final concentration) of purified bacterially expressed Aurora A is assayed in the presence of the appropriate inhibitors (MLN8054, Alisertib), using Histone H3 as substrate for 20 min at 30°C in the presence of 100 μM [γ-32P] ATP. For Aurora A/TPX2 assays, 50 ng of a TPX2 [1-43] peptide, representing a 2-fold molar excess over Aurora A, is included. The Aurora A/TPX2 complex is preformed in kinase reactions prior to subsequent addition of inhibitors and ATP.

MM cell lines are incubated with DMSO or Alisertib (0.125-0.5 μM) in combination with conventional anti-MM agents melphalan (2.5-5 μM), doxorubicin (50-100 nM), or dexamethasone (50-100 nM); and with novel anti-MM agents bortezomib (2.5-5 nM) or lenalidomide (0.5-1 μM) for 72 hours. Cell viability is measured by MTT assay. The combination index (CI) is determined by isobologram analysis using CalcuSyn software, Version 2.0 (CI < 1 indicates synergistic effect; CI=1, additive effect; and CI > 1, no significant combination effect).

Mice are irradiated (200 cGy), and then 5×106 MM1.S cells are inoculated subcutaneously in the right flank. When tumor growth is measurable (2 weeks after the injection), mice are assigned into 4 groups (10 mice each) receiving vehicle orally (100 μL of 10% 2-hydroxypropyl-β-cyclodextrin/1% sodium bicarbonate) or Alisertib (7.5 mg/kg, 15 mg/kg, and 30 mg/kg in a final formulation in 10% 2-hydroxypropyl-β-cyclodextrin/1% sodium bicarbonate) for 21 consecutive days. The maximal tolerated dose of Alisertib in most mouse strains (continuous dosing for 21 days) is approximately 20 mg/kg twice a day (40 mg/kg per day). Tumor volumes are measured by a Vernier caliper every alternate day and calculated using the following formula: length×width2×0.5. Mice are killed at the end of the treatment, 2 hours after the last treatment, or when tumor reaches 2 cm3; tumors are immediately collected from mice and evaluated for induction of apoptosis and cell death by TdT-mediated dUTP nick end labeling (TUNEL) assay.

[1]. Güllü G, et al. A novel Aurora-A kinase inhibitor MLN8237 induces cytotoxicity and cell-cycle arrest in multiple myeloma Blood June 24, 2010 vol. 115 no. 25 5202-5213

[2]. Sloane DA, et al. Drug-Resistant Aurora A Mutants for Cellular Target Validation of the Small Molecule Kinase Inhibitors MLN8054 and MLN8237 ACS Chem. Biol., 2010, 5 (6), pp 563-576

[3]. Manfredi MG, et al. Characterization of Alisertib (MLN8237), an investigational small-molecule inhibitor of aurora A kinase using novel in vivo pharmacodynamic assays.Clin Cancer Res. 2011 Dec 15;17(24):7614-7624.

[4]. Yayi Feng, et al. Targeting aurora a kinase (AURKA) and P21-activated kinase 1 (PAK1) in hormone receptor-positive breast cancer. College of Medicine. Drexel University. November 2016.

Tozasertib | AT9283 | Barasertib (AZD1152-HQPA) | danusertib | Hesperadin | Reversine | MLN8054 | GSK1070916 | AMG-900 | KW-2449 | TAK-632 | TAK-901 | MK-5108(VX-689) | pf-03814735 | ENMD-2076