281 semi preparative hplc system

Example 57

To a solution of 58a-int (80 mg, 149.42 umol) in H₂O (800 uL) was added HCl (12 N, 12.45 uL) and AcOH (8.97 mg, 149.42 umol, 8.54 uL). The mixture was stirred at 70° C. for 12 hrs. LC-MS showed 57a-int was consumed completely and one main peak with desired MS for Example 57 was detected. The reaction mixture was concentrated and was purified by prep-HPLC to give Example 57.

HPLC purification conditions: Gilson 281 semi-preparative HPLC system; Mobile phase A: TFA/H₂O=0.075% v/v; B: AcN (gradient % B at T=0: 14%; at T=10.5 min: 34%); Column: Boston Green ODS 150×30 5 u; Flow rate: 25 mL/min; Monitor wavelength: 220 & 254 nm.

¹H NMR (400 MHz, METHANOL-d₄): δ ppm 7.32-7.39 (m, 3H), 7.22 (d, J=8.60 Hz, 2H), 6.82-6.85 (m, 2H), 4.87-4.89 (m, 1H), 4.22 (s, 2H), 4.00-4.05 (m, 2H), 3.47 (br t, J=6.73 Hz, 2H), 3.21 (dd, J=14.22, 5.40 Hz, 1H), 3.12-3.15 (m, 3H), 2.96 (dd, J=14.22, 9.15 Hz, 1H), 1.85-1.93 (m, 4H); LCMS (ESI+): m/z=521.0 (M+H)⁺, RT: 2.3 min; HPLC purity: 97.2%, RT: 5.5 min; Chiral SFC: purity: 100%, ee value: 100%, RT: 2.8 min.

Example 20

To a solution of 20a-int (150 mg, 305.86 umol) in THF (2 mL)/H₂O (2 mL) was added LiOH. H₂O (12.83 mg, 305.86 umol). This was stirred for 12 hrs at 20° C. after which LCMS showed the desired product was present and the reaction complete. The solvent was removed and the crude material was purified by prep-HPLC to give Example 20.

HPLC purification conditions: Gilson 281 semi-preparative HPLC system; Mobile phase A: TFA/H₂O=0.075% v/v; B: AcN (gradient % B at T=0: 20%; at 11.5 min: 40%); Column: Boston Green ODS 150×30 5 u; Flow rate: 25 mL/min; Monitor wavelength: 220 & 254 nm.

¹H NMR (400 MHz, METHANOL-d₄): 7.56 (d, J=7.2 Hz, 1H), 7.35-7.41 (m, 3H), 6.60 (d, J=7.2 Hz, 1H), 5.52-5.63 (m, 1H), 4.60-4.64 (m, 1H), 3.44-3.46 (m, 2H), 2.78-2.79 (m, 2H), 2.68-2.76 (m, 2H) 2.64-2.66 (m, 1H), 2.05-2.08 (m, 2H), 1.89-1.92 (m, 2H), 1.65-1.67 (m, 2H), 1.43-1.46 ppm (m, 2H); LCMS (ESI+): m/z=476.0 (M+H)+, RT=2.3 min; HPLC: RT=4.9 min, purity: 90.9%; Chiral SFC purity: 100%, e.e. value: 100%, RT=3.3 min.

Example 23

To 23c-int (0.4 g) in THF (10 mL)/H₂O (10 mL) was added LiOH. H₂O (0.16 g, 3.78 mmol). The mixture was stirred for 2 h at 30° C. then was concentrated and purified by prep-HPLC to give Example 23.

HPLC purification conditions: Gilson 281 semi-preparative HPLC system; Mobile phase A: TFA/H₂O=0.075% v/v; B: AcN (gradient % B at T=0: 10%; at T=12 min: 30%); Column: Luna C18 100×30 5 u; Flow rate: 20 mL/min; Monitor wavelength: 220 & 254 nm.

¹H NMR (400 MHz, CHLOROFORM-d): δ ppm 2.03-1.97 (m, 2H) 2.59-2.49 (m, 3H) 3.21 (t, J=10.4 Hz, 2H) 3.89 (d, J=5.2 Hz, 3H) 5.12-5.07 (m, 1H) 5.42 (s, 1H) 6.38 (d, J=7.6 Hz, 1H) 6.69-6.63 (m, 2H) 6.77 (d, J=8.8 Hz, 2H) 7.15 (d, J=8.8 Hz, 2H) 7.28-7.22 (m, 3H) 7.79-7.74 (m, 2H) 9.67 (s, 1H); LCMS (ESI+): m/z=514.0 (M+H)⁺, RT: 2.2 min; HPLC purity: 95.3%, RT: 6.2 min; Chiral SFC purity: 100%, ee value: 100%, RT: 3.4 min.

Example 54

To 54c-int (65 mg, 133.07 umol) in THF (3 mL)/H₂O (1 mL) was added LiOH.H₂O (11.17 mg, 266.14 umol). The mixture was stirred at 25° C. for 12 hours. LCMS showed 54c-int was consumed and one main peak for Example 54 was detected. The mixture was adjusted to pH 6 with aqueous HCl (1N) then concentrated and purified by prep-HPLC to give Example 54.

HPLC purification conditions: Gilson 281 semi-preparative HPLC system; Mobile phase A: 10 mM NH₄HCO₃ in H₂O; B: AcN (gradient % B at T=0: 30%; at T=12 min: 50%); Column: YMC-Actus Triart C18 150×30 5 u; Flow rate: 25 mL/min; Monitor wavelength: 220 & 254 nm.

¹H NMR (400 MHz, METHANOL-d₄): δ ppm 7.32-7.47 (m, 3H), 4.62 (dd, J=9.15, 4.74 Hz, 1H), 3.51 (br t, J=5.51 Hz, 2H), 3.20-3.29 (m, 4H), 2.13-2.22 (m, 2H), 1.88-2.00 (m, 1H), 1.71-1.84 (m, 1H), 1.57-1.68 (m, 2H), 1.48-1.56 (m, 1H), 1.48-1.49 (m, 1H), 1.38 (br s, 8H); LCMS (ESI+): m/z=474.0 (M+H)⁺, RT: 2.5 min; HPLC purity: 97.6%, RT: 6.3 min; Chiral SFC purity: 99.1%, ee value: 98.2%, RT: 3.0 min.

Example 22

To 22f-int (100 mg, 185.37 umol) in THF (5 mL) was added LiOH (13.32 mg, 556.12 umol) and H₂O (2 mL), then the mixture was stirred at 40° C. for 12 hrs. After this time LCMS showed the reaction was complete. The mixture was concentrated then purified by prep-HPLC to give Example 22.

HPLC purification conditions: Gilson 281 semi-preparative HPLC system; Mobile phase A: TFA/H₂O=0.075% v/v; B: AcN (gradient % B at T=0: 12%; at 10.5 min: 34%); Column: Boston Green ODS 150×30 5 u; Flow rate: 25 mL/min; Monitor wavelength: 220 & 254 nm.

¹H NMR (400 MHz, METHANOL-d₄): 6 ppm 7.63 (d, J=7.2 Hz, 1H), 7.32-7.36 (m, 3H), 7.17 (d, J=8.4 Hz, 2H), 6.83 (d, J=7.5 Hz, 1H), 6.50 (d, J=8.4 Hz, 2H), 4.81 (br s, 1H), 4.16-4.23 (m, 2H), 3.90 (br d, J=2.8 Hz, 2H), 3.47-3.53 (m, 2H), 3.27-3.28 (m, 1H), 3.17 (dd, J=14.0, 5.52 Hz, 1H), 2.92 (dd, J=14.2, 9.2 Hz, 1H), 2.83 (t, J=6.4 Hz, 2H), 1.92-2.00 (m, 2H); LCMS (ESI+): m/z=525.0 (M+H)⁺, RT: 2.5 min; HPLC purity: 90.5%, RT: 6.2 min; Chiral SFC purity: 87.3%, ee value: 74.5%, RT: 1.3 min.

Example 53

A mixture of compound 53d-int (100 mg, 200.23 umol) and LiOH.H₂O (8.40 mg, 200.23 umol) in THF (5 mL)/H₂O (1 mL) was stirred at 20° C. for 1 hr. LCMS showed desired MS; the mixture was concentrated and the residue was purified by prep-HPLC (TFA) to give Example 53.

HPLC purification conditions: Gilson 281 semi-preparative HPLC system; Mobile phase A: TFA/H₂O=0.075% v/v; B: AcN (gradient % B at T=0: 25%; at T=12 min: 55%); Column: YMC-Actus Triart C18 150×30 5 u; Flow rate: 25 mL/min; Monitor wavelength: 220 & 254 nm.

¹H NMR (400 MHz, METHANOL-d₄): δ ppm 7.30-7.50 (m, 3H), 4.62 (dd, J=9.26, 4.63 Hz, 1H), 3.59-3.74 (m, 4H), 3.33 (s, 2H), 3.11 (s, 3H), 1.87-2.01 (m, 1H), 1.72-1.85 (m, 1H), 1.60-1.71 (m, 2H), 1.47-1.59 (m, 2H), 1.38 (br s, 8H); LCMS (ESI+): m/z=485.2 (M+H)⁺, RT: 2.4 min; HPLC purity: 90.6%, RT: 5.8 min; Chiral SFC purity: 100%, ee value: 100%, RT: 2.6 min.

Example 51

To 51c-int (100 mg, 211.69 umol) in THF (5 mL) and H₂O (2 mL) was added LiOH.H₂O (17.77 mg, 423.38 umol); the mixture was stirred at 25° C. for 2 hours. LCMS showed 51c-int was consumed completely and one main peak with desired MS was detected. The mixture was concentrated and adjusted to pH 6 with aq. HCl (1N) and was concentrated in vacuum. The residue was purified by prep-HPLC to give Example 51.

HPLC purification conditions: Gilson 281 semi-preparative HPLC system; Mobile phase A: 10 mM NH₄HCO₃ in H₂O; B: AcN (gradient % B at T=0: 25%; at T=12 min: 45%); Column: YMC-Actus Triart C18 150×30 5 u; Flow rate: 25 mL/min; Monitor wavelength: 220 & 254 nm.

¹H NMR (400 MHz, METHANOL-d₄): 6 ppm 7.33-7.47 (m, 3H), 4.56 (dd, J=8.27, 4.74 Hz, 1H), 3.21-3.30 (m, 6H), 1.86-2.00 (m, 3H), 1.71-1.84 (m, 1H), 1.45-1.58 (m, 4H), 1.26-1.42 (m, 7H), 1.25-1.26 (m, 1H); LCMS (ESI+): m/z=458.0 (M+H)⁺, RT: 2.5 min; HPLC purity: 100%, RT: 6.5 min; Chiral SFC purity: 100%, ee value: 100%, RT: 3.0 min.