Reversine, a small synthetic purine analogue (2, 6-disubstituted purine), is a potent inhibitior of Aurora A/B/C(IC50s=150-500 nM).

Aurora A:150-500 nM, Aurora B:150-500 nM, Aurora C:150-500 nM

在携带U14肿瘤的小鼠体内,腹腔注射Reversine（10 mg/kg）能够消退肿瘤细胞.

在稳定转染的中国仓鼠卵巢细胞中,Reversine能够竞争性抑制毛喉素刺激的cAMP产生。在HCT116细胞中,Reversine抑制Aurora靶点,及组蛋白H3的磷酸化。在原代人肿瘤样品中,Reversine能够抑制白血病细胞集落的形成。

Radioligand Binding Assays: Each tube in the A3 AR competitive binding assay contains 100 μL of membrane suspension (20 μg of protein), 50 μL of [125I]4-amino-3-iodobenzyl)adenosine-5′-N-methyluronamide (0.5 nM), and 50 μL of increasing concentrations of the test ligands in Tris-HCl buffer (50 mM, pH 7.4) containing 10 mM MgCl2 and 1 mM EDTA. Nonspecific binding is determined using 10 mM 5′-N-ethylcarboxamidoadenosine in the buffer. The mixtures are incubated at 25°C for 60 min. Binding reactions are terminated by filtration through Whatman GF/B filters under reduced pressure using a MT-24 cell harvester. Filters are washed three times with 9 mL of ice-cold buffer. Radioactivity is determined using a Beckman γ-counter, and the percent inhibition is calculated.

Cell viability of different tumor cell lines is assessed using ATPlite 1step. Briefly, 2 × 104?cells for each well are plated in a 96-well plate in presence of crescent quantity of reversine. After 72 h, the plates are recovered and 100 μL ATPlite solution is added to each well. The plates are shaken for 2 min at 700 rpm and luminescence is measured using EnVision Multilabel plate reader. Each sample is analyzed in triplicate.(Only for Reference)