2-Dicyclohexylphosphino-2',4',6'-triisoprophylbiphenyl → HCID 54765114, compound

Temperature

100 °C

To a stirred solution of dicyclohexyl(2′,4′,6′-triisopropylbiphenyl-2-yl)phosphine (0.018 g, 0.038 mmol), 4-(3-bromo-1,2,4-thiadiazol-5-yl)morpholine (0.059 g, 0.24 mmol), 7-fluoro-3-methyl-2-(pyridin-2-yl)quinolin-4-amine (0.06 g, 0.24 mmol) and Pd2 dba3 (0.009 g, 9.48 μmol) in toluene (2.4 mL) was added sodium tert-butoxide (0.057 g, 0.592 mmol). The reaction mixture was heated to 100° C. and stirred for 1 hour. The crude reaction mixture was filtered through a plug of alumina eluting with (dichloromethane/methanol; 3/1), the filtrate was concentrated in vacuo. The crude product was purified by column chromatography on alumina (0-60% ethyl acetate/hexanes) to give the desired product. The desired product was further purified with HPLC (10-90% of 0.1% TFA acetonitrile solution in 0.1% TFA water solution.) The desired fractions were concentrated then diluted with ethyl acetate. After washing twice with saturated aq. sodium bicarbonate solution, the solvent was removed under reduced pressure to yield pure product N-(7-fluoro-3-methyl-2-(pyridin-2-yl)quinolin-4-yl)-5-morpholino-1,2,4-thiadiazol-3-amine. 1H NMR (400 MHz, CD2Cl2) δ ppm 8.65 (1H, d, J=3.9 Hz), 8.03 (1H, dd, J=9.2, 5.9 Hz), 7.94 (1H, td, J=7.7, 1.8 Hz), 7.74 (1H, d, J=7.8 Hz), 7.67 (1H, dd, J=10.0, 2.7 Hz), 7.39-7.49 (1H, m), 7.32 (1H, ddd, J=9.3, 8.2, 2.4 Hz), 3.71-3.83 (4H, m), 3.37-3.50 (4H, m), 2.28 (3H, s). Mass Spectrum (ESI) m/e=423.2 (M+1). Biological Assays Recombinant Expression of PI3Ks Full length p110 subunits of PI3k α, β and δ, N-terminally labeled with polyHis tag, were coexpressed with p85 with Baculo virus expression vectors in sf9 insect cells. P110/p85 heterodimers were purified by sequential Ni-NTA, Q-HP, Superdex-100 chromatography. Purified α, β and δ isozymes were stored at −20° C. in 20 mM Tris, pH 8, 0.2M NaCl, 50% glycerol, 5 mM DTT, 2 mM Na cholate. Truncated PI3Kγ, residues 114-1102, N-terminally labeled with polyHis tag, was expessed with Baculo virus in Hi5 insect cells. The γ isozyme was purified by sequential Ni-NTA, Superdex-200, Q-HP chromatography. The γ isozyme was stored frozen at −80° C. in NaH2PO4, pH 8, 0.2M NaCl, 1% ethylene glycol, 2 mM β-mercaptoethanol.

1. customThe crude reaction mixture
2. filtrationwas filtered through a plug of alumina eluting with (dichloromethane/methanol; 3/1)
3. concentrationthe filtrate was concentrated in vacuo
4. customThe crude product was purified by column chromatography on alumina (0-60% ethyl acetate/hexanes)

ORD record

ord-c6f75ca2b8234f35b4832e77776539e4

Source dataset

uspto-grants-2014_04

DOI

10.6084/m9.figshare.5104873.v1

Patent number

US08686137B2