Reaction Steps
Ethyl cis-4-(7-{[2-({4-[(diethoxyphosphoryl)methyl]-2-methoxyphenyl}amino)-5-(trifluoromethyl)pyrimidin-4-yl]amino}-2-methyl-1-oxo-2,3-dihydro-1H-isoindol-4-yl)cyclohexanecarboxylate (Example 87, 64.0 mg, 0.087 mmol) was taken up in THF (0.50 mL, 6.2 mmol) and MeOH (0.50 mL, 12 mmol). This was treated with a solution of Lithium hydroxide, monohydrate (18.3 mg, 0.44 mmol) in H2O (0.50 mL, 28 mmol) and allowed to stir at rt for 30 hours, during which, the reaction mixture transformed from a thick slurry to a homogenous solution. The reaction was concentrated in vacuo to a solid. The crude product was purified by CombiFlash®Rf 4X Organic Purification System [12 g RediSep® Normal-phase GOLD Silica Flash Column, dried loaded, elution gradient: 0→7.5% MeOH in 1:1 EtOAc/DCM] to afford 4.0 mg (6% yield) of the desired product. 1H NMR (400 MHz, DMSO-d6) δ ppm 8.95 (s, 1 H), 8.38 (s, 1 H), 7.52 (br. s., 1 H), 7.19 (br. s., 1 H), 7.05 (s, 1 H), 6.90 (d, J=8.59 Hz, 1 H), 4.50 (s, 2 H), 3.95-4.06 (m, 4 H), 3.77 (s, 3 H), 3.21-3.29 (m, 2 H), 3.08 (s, 3 H), 2.67 (br. s., 1 H), 2.60 (br. s., 1 H), 2.15 (d, J=7.83 Hz, 2 H), 1.55-1.69 (m, 6 H), 1.21 (t, 6 H). MS (ES+): m/z=706.81 [MH+]. HPLC: tR=1.55 min (UPLC TOF, polar—3 min). Example 89 may be further prepared as a salt. Non-limiting examples include the sodium, potassium, ammonium, 1,3-dihydroxy-2-(hydroxymethyl)propan-2-aminium [tromethamine], lysine and choline salts. This can be achieved by combining the compound with sodium hydroxide, potassium hydroxide, ammonium hydroxide, or 2-amino-2-(hydroxymethyl)propane-1,3-diol [tromethamine], lysine or choline by mixing of solids where appropriate or by combining solutions of known concentrations of either or both components. Known techniques include filtration of a suspension (if the salt precipitates) or concentration or lyophilization of the salt solution. The compound may be formulated as a solution using, for example, a combination of water, a cyclodextrin such as hydroxypropyl-β-cyclodextrin (HPBCD), a surfactant such as sodium lauryl sulfate (SLS), a PEG ether such as PEG-400, propylene glycol (PG) and a buffer to modulate the pH such as a bicarbonate, carbonate or hydroxide salt (or combination thereof). Some non-limiting examples of this are aqueous mixtures containing 5-10% SLS buffered to pH 9.5, 10% SLS+30% PEG400+15% PG, 40% HPBCD, and 20-40% HPBCD buffered to pH 9.5. The skilled artisan will realize there are other excipients and combinations including solid, suspension and other solution forms.
