CYT997 (Lexibulin)

854074-39-8

917111-46-7

917111-44-5

Under nitrogen protection, 2-chloro-5-methyl-N-[(1S)-1-pyridin-3-ylbutyl]pyrimidin-4-amine (277 mg, 1.0 mmol), 4-{[(ethylamino)carbonyl]amino}-3-methoxybenzeneboronic acid pinacol ester (416 mg, 1.3 mmol), and tetrakis(triphenylphosphine)palladium(0) (116 mg, 0.1 mmol) were were dissolved in a mixture of toluene-n-propanol (12 mL, 3:1, v/v) and 2M aqueous sodium carbonate (750 μL, 1.5 mmol) was added. The reaction mixture was heated at 100 °C for 17 h with vigorous stirring. After the reaction was completed, it was cooled to room temperature, diluted by adding ethyl acetate (25 mL), and the mixture was washed sequentially with water (6 × 15 mL), brine (20 mL), and dried over anhydrous sodium sulfate. The solvent was removed by concentration under reduced pressure to obtain the crude product. The crude product was purified by column chromatography using dichloromethane-methanol-ammonia (93:7:1, v/v/v) as eluent to give the target product (110 mg, 57% yield).1H NMR (CDCl3) δ 0.99 (t, 3H, J = 7.4 Hz, CH3), 1.18 (t, 3H, J = 7.2 Hz, CH3), 1.33- 1.60 (m, 2H, CH2), 1.82-2.02 (m, 2H, CH2), 2.11 (s, 3H, Ar-Me), 3.25-3.39 (m, 2H, CH2), 3.87 (s, 3H, OMe), 4.80-4.96 (m, 2H, 2 × NH), 5.26-5.36 (m, 1H, CH), 6.98 (br s, 1H, CH), 4.80-4.96 (m, 1H, CH), 5.26-5.36 (m, 1H, CH), 6.98 (br s, 1H, J = 7.2 Hz, CH 6.98 (br s, 1H, Ar-NHCONH), 7.22-7.28 (m, 1H, ArH), 7.67-7.72 (m, 2H, ArH), 7.83-7.88 (m, 1H, ArH), 8.05 (d, 1H, J = 0.8 Hz, ArH), 8.10 (d, 1H, J = 8.2 Hz, ArH), 8.47 (dd, 1H, J = 8.2 Hz, ArH), 8.47 (dd, 1H, J = 8.2 Hz, ArH), 8.47 (dd, 1H, J = 8.2 Hz, ArH) 8.47 (dd, 1H, J = 6.6, 1.8 Hz, ArH), 8.69 (d, 1H, J = 2.0 Hz, ArH).

Treatment of A549 cells with CYT997 (1 μM) for 24 hours induced rapid reorganization of microtubules, including disruption of the existing microtubule network and accumulation of some cytoplasmic tubulin in plaques, resulting in significant changes in cell morphology, including Adherent cells are lost and cells are reduced. CYT997 is toxic to 16 cancer cells with IC50 ranging from 9 nM in HepG2 cells to 101 nM in KHOS/NP cells. CYT997 effectively acts on HCT15 cells, has a multi-drug resistance mechanism Pgp (MDR consistent with CYT997 destroys cellular tubulin, CYT997 effectively inhibits proliferation, induces cell cycle arrest, and induces apoptosis in human myeloid cell lines (HMCLs) and primary MM cells die.