Synergi c18 150

Example 56

[Figure (not displayed)]

To a solution of thiophene-2-carboxylic acid (22.93 mg, 178.90 μmol), HATU (68.02 mg, 178.90 μmol) and DIPEA (84.08 mg, 650.55 μmol) in DCM (1.0 mL) was added (2S)-2-amino-4-methylsulfanyl-N-(4-phenylthiazol-2-yl)butanamide (50.00 mg, 162.64 μmol) and the mixture was stirred at 20° C. for 2 hr. The reaction mixture was concentrated under reduced pressure to give a residue. The residue was purified by prep-HPLC (column: Phenomenex Synergi C18 150*25*10 μm; mobile phase: [water (0.1% TFA)-ACN]; B %: 50%-80%, 9 min) and lyophilized to give compound 58 as a white solid.

¹H NMR (400 MHz, CDCl₃) δ=7.69 (d, J=7.2 Hz, 2H), 7.62-7.61 (m, 1H), 7.48 (d, J=4.8 Hz, 1H), 7.40-7.36 (m, 2H), 7.34-7.30 (m, 1H), 7.26-7.24 (d, J=8.0 Hz, 1H), 7.07 (s, 1H), 7.06-7.03 (m, 1H), 5.00-4.95 (m, 1H), 2.68-2.63 (m, 2H), 2.29-2.20 (m, 2H), 2.10 (s, 3H) ppm.

LCMS (ESI) m/z: [M+H]⁺=418.0

Chiral HPLC: Amycoat-MeOH(DEA)-40-7 min-3 mL-35T, 3.772 min.

Example 29

[Figure (not displayed)]

¹H NMR (400 MHz, DMSO-d₆) δ=1.31 (dq, J=15.10, 6.46 Hz, 1H), 1.69-1.82 (m, 1H), 2.18-2.26 (m, 1H), 2.31 (s, 3H), 4.58 (br d, J=3.79 Hz, 2H), 4.91-5.17 (m, 1H), 5.42 (br s, 1H), 7.31 (d, J=8.07 Hz, 1H), 7.42 (s, 1H), 7.72 (s, 1H), 8.06 (d, J=8.07 Hz, 1H), 8.33 (s, 1H), 12.70 (br s, 1H); LCMS (electrospray) m/z 358.3 (M+H)+.

Synthetic Method I

Example 113

[Figure (not displayed)]

To a solution of Example 29 (380 mg, 1.06 mmol, 1 eq) in DCM (5 mL) was added DAST (685.52 mg, 4.25 mmol, 561.90 μL, 4 eq) at 0° C. The mixture was stirred at 30° C. for 3 hr under N2 atmosphere. The reaction mixture was diluted with DCM (5 mL), and then the resulting organic phase was washed with saturated sodium bicarbonate aqueous solution (10 mL) and brine (10 mL), dried over sodium sulfate, filtered and the filtrate was concentrated in vacuum to give a residue. The residue was purified by pre-HPLC (column: Phenomenex Synergi C18 150*25*10 um; mobile phase: [water (0.1% TFA)-ACN]; B %: 20%-50%, 10 min).

¹H NMR (400 MHz, DMSO-d₆) δ=1.32 (ddt, J=12.84, 8.99, 6.33, 6.33 Hz, 1H), 1.69-1.82 (m, 1H), 2.20-2.28 (m, 1H), 2.36 (s, 3H), 4.93-5.17 (m, 1H), 5.46 (s, 1H), 5.57 (s, 1H), 7.48 (dd, J=8.31, 1.83 Hz, 1H), 7.53 (s, 1H), 7.84 (d, J=8.31 Hz, 1H), 8.07 (d, J=1.59 Hz, 1H), 8.48 (s, 1H), 12.77 (br s, 1H); LCMS (electrospray) m/z 360.0 (M+H)+.

Synthetic Method Z

Example 29

[Figure (not displayed)]

To a solution of 1-[[(2S,3S,4S)-3-ethyl-4-fluoro-5-oxo-pyrrolidin-2-yl]methoxy]-4-[2-(4-formyl cyclohexyl)ethynyl]-7-methoxy-isoquinoline-6-carboxamide (50.0 mg, 100 umol, Intermediate AFM) in methanol (2 mL) was added NH₄OAc (1.56 g, 20.1 mmol). After the reaction mixture was stirred at 20° C. for 3 hr, NaBH₃CN (9.51 mg, 151 umol) was added. The reaction mixture was stirred at 20° C. for 16 hrs. On completion, the reaction mixture was quenched by water (0.2 mL) and concentrated in vacuo. The residue was purified by prep-HPLC (column: Phenomenex Synergi C18 150*25*10 um; mobile phase: [water (0.225% FA)-ACN]; B %: 15%-35%, 10 min) to give the title compound (10.5 mg, 18% yield, FA salt) as off-white solid. ¹H NMR (400 MHz, DMSO-d₆) δ 8.87 (s, 1H), 8.40 (s, 1H), 8.02 (s, 1H), 7.95-7.68 (m, 3H), 7.33-6.42 (m, 2H), 4.99-4.81 (m, 1H), 4.53 (dd, J=3.2, 11.2 Hz, 1H), 4.26 (dd, J=6.4, 11.2 Hz, 1H), 4.12-4.05 (m, 1H), 3.98 (s, 3H), 2.66-2.62 (m, 1H), 2.62-2.59 (m, 2H), 2.58-2.52 (m, 2H), 2.17-2.02 (m, 2H), 1.90-1.76 (m, 2H), 1.68-1.55 (m, 2H), 1.55-1.40 (m, 3H), 1.10-0.97 (m, 4H); LC-MS (ESI⁺) m/z 497.2 (M+H)⁺.

Example 687

[Figure (not displayed)]

To a solution of N5-[1-[2-[2-(2-aminoethoxy)ethoxy]ethyl]pyrazol-4-yl]-1-methyl-N7-(4-morpholinocyclohexyl)pyrazolo[4,3-d]pyrimidine-5,7-diamine (60.0 mg, 106 umol, HCl, Intermediate UT) and 2-(2,6-dioxo-3-piperidyl)-4-fluoro-isoindoline-1,3-dione (29.3 mg, 106 umol, Intermediate R) in DMF (2 mL) was added DIPEA (68.6 mg, 530 umol, 92.4 uL). The mixture was stirred at 90° C. for 16 hours. On completion, the reaction mixture was concentrated in vacuo. The residue was purified by prep-HPLC (column: Phenomenex Synergi C18 150*25*10 um; mobile phase: [water (0.225% FA)-ACN]; B%: 9%-30%, 7 min) to give the title compound (13.2 mg, 15% yield, FA) as yellow solid. ¹H NMR (400 MHz, DMSO-d₆) δ 11.09 (s, 1H), 8.55 (s, 1H), 7.92 (s, 1H), 7.56 (s, 1H), 7.54-7.43 (m, 2H), 7.05-6.96 (m, 2H), 6.55 (t, J=5.6 Hz, 1H), 6.43 (d, J=7.6 Hz, 1H), 5.04 (dd, J=5.2, 12.8 Hz, 1H), 4.17 (t, J=5.2 Hz, 2H), 4.12 (s, 3H), 4.10-4.04 (m, 1H), 3.74 (t, J=5.2 Hz, 2H), 3.57-3.51 (m, 14H), 2.94-2.78 (m, 2H), 2.63-2.58 (m, 1H), 2.57-2.54 (m, 1H), 2.28-2.21 (m, 1H), 2.13-1.82 (m, 6H), 1.56-1.45 (m, 2H), 1.39-1.29 (m, 2H); LC-MS (ESI⁺) m/z 785.4 (M+H)⁺.

Example 5

[Figure (not displayed)]

Example 58

[Figure (not displayed)]

To a solution of Compound 1 (200 mg, 552.14 μmol, 1 eq) in ACETONE (5 mL) and H₂O (5 mL) was added NaIO₄ (708.59 mg, 3.31 mmol, 183.57 μL, 6 eq) and NH₄HCO₃ (261.90 mg, 3.31 mmol, 272.81 μL, 6 eq). The mixture was stirred at 40° C. for 1 hr. The reaction mixture was concentrated under reduced pressure to remove solvent. The residue was diluted with H₂O (20 mL), then the mixture was extracted with ethyl acetate (20 mL*3). The combined organic layers were washed with brine (20 mL*2), dried over Na₂SO₄, filtered and concentrated under reduced pressure to give a residue. The residue was purified by pre-HPLC (column: Phenomenex Synergi C18 150*25*10 um; mobile phase: [water (0.05% HCl)-ACN]; B %: 15%-41%, 8 min). Example 58 (58 mg, 172.24 μmol, 31% yield, 94% purity, HCl) was obtained as a light yellow solid.

¹H NMR (400 MHz, METHANOL-d₄) δ=1.26-1.39 (m, 1H), 1.81-1.94 (m, 1H), 2.19 (dtd, J=9.19, 6.87, 6.87, 4.34 Hz, 1H), 4.87-5.18 (m, 1H), 7.69-7.75 (m, 1H), 7.81 (br d, J=7.82 Hz, 1H), 8.22 (s, 1H); LCMS (electrospray) m/z 281.2 (M+H)+.

Synthetic Method P

Example 2

[Figure (not displayed)]

To a solution of methyl 2-[4-(hydroxymethyl)cyclohexyl]-6-[[6-(trifluoromethyl)pyridine-2-carbonyl]amino]-1,3-benzothiazole-5-carboxylate (120 mg, 243 umol, synthesized via Step 1 of Intermediate BAX) in THF (10 mL) was added MeMgBr (3 M, 405 uL) and the mixture was stirred at 0° C. for 2 hrs. On completion, the reaction mixture was quenched by addition 10 mL sat. NH₄Cl at 0° C., and then diluted with 50 mL water and extracted with EA (50 mL×3). The combined organic layers were washed with 100 mL brine, dried over Na₂SO₄, filtered and concentrated in vacuo to give a residue. The residue was purified by pre-HPLC (column: Phenomenex Synergi C18 150*25*10 um; mobile phase: [water (0.225% FA)-ACN]; B %: 44/6-74%, 10 min) to give the title compound (80.0 mg, 65% yield) as white solid. ¹H NMR (400 MHz, DMSO-d₆) δ 12.56 (s, 1H), 9.07 (s, 1H), 8.51-8.45 (m, 1H), 8.39 (t, J=8.0 Hz, 1H), 8.20 (d, J=7.6 Hz, 1H), 7.94-7.88 (m, 1H), 6.08 (s, 1H), 4.46 (t, J=5.2 Hz, 1H), 3.28 (t, J=5.6 Hz, 2H), 3.10-3.00 (m, 1H), 2.19 (d, J=11.2 Hz, 2H), 1.94-1.84 (m, 2H), 1.64 (s, 6H), 1.61-1.53 (m, 2H), 1.50-1.40 (m, 1H), 1.19-1.06 (m, 2H); LC-MS (ESI⁺) m/z 494.0 (M+1)⁺.

Example 687

[Figure (not displayed)]

To a solution of N5-[1-[2-[2-(2-aminoethoxy)ethoxy]ethyl]pyrazol-4-yl]-1-methyl-N7-(4-morpholinocyclohexyl)pyrazolo[4,3-d]pyrimidine-5,7-diamine (60.0 mg, 106 umol, HCl, Intermediate UT) and 2-(2,6-dioxo-3-piperidyl)-4-fluoro-isoindoline-1,3-dione (29.3 mg, 106 umol, Intermediate R) in DMF (2 mL) was added DIPEA (68.6 mg, 530 umol, 92.4 uL). The mixture was stirred at 90° C. for 16 hours. On completion, the reaction mixture was concentrated in vacuo. The residue was purified by prep-HPLC (column: Phenomenex Synergi C18 150*25*10 um; mobile phase: [water (0.225% FA)-ACN]; B %: 9%-30%, 7 min) to give the title compound (13.2 mg, 15% yield, FA) as yellow solid. ¹H NMR (400 MHz, DMSO-d₆) δ 11.09 (s, 1H), 8.55 (s, 1H), 7.92 (s, 1H), 7.56 (s, 1H), 7.54-7.43 (m, 2H), 7.05-6.96 (m, 2H), 6.55 (t, J=5.6 Hz, 1H), 6.43 (d, J=7.6 Hz, 1H), 5.04 (dd, J=5.2, 12.8 Hz, 1H), 4.17 (t, J=5.2 Hz, 2H), 4.12 (s, 3H), 4.10-4.04 (m, 1H), 3.74 (t, J=5.2 Hz, 2H), 3.57-3.51 (m, 14H), 2.94-2.78 (m, 2H), 2.63-2.58 (m, 1H), 2.57-2.54 (m, 1H), 2.28-2.21 (m, 1H), 2.13-1.82 (m, 6H), 1.56-1.45 (m, 2H), 1.39-1.29 (m, 2H); LC-MS (ESI⁺) m/z 785.4 (M+H)⁺.