High Performance Liquid Chromatography:Article Title: Substituted aminothiazoles as DGKzeta inhibitors for immune activation
Article Snippet: Preparative chiral HPLC Instrument: Sepiatec: Prep SFC100; Column: Chiralpak IA 5μ 250 × 30 mm; eluent A: CO2; eluent B: 2-propanol + 0.4 vol % diethylamine; isocratic: 25% B; flow: 100 mL/min; temperature: 40° C.; BPR: 150 bar; UV: 280 nm Analytical chiral HPLC: Rt = 4.57 min Instrument: Agilent: 1260, Aurora SFC-module; column: Chiralpak IA 5μ 100 × 4.6 mm; eluent A: CO2; eluent B: 2-propanol + 0.4 vol % diethylamine, isocratic: 25% B; flow: 4 mL/min; temperature: 37.5° C.; BPR: 100 bar; UV: 280 nm 74 Intermediate 92; rac-2- bromo- propanamide 1H-NMR (400 MHz, DMSO-d6): δ ppm = 1.18 (d, J = 7.35 Hz, 3 H), 5.05 (q, J = 7.18 Hz, 1 H), 7.28 (s, 1 H), 7.32 (td, J = 8.30, 2.15 Hz, 1 H), 7.37-7.46 (m, 4 H), 7.47-7.56 (m, 4 H), 7.60 (s, 1 H), 7.85-8.35 (m, 2 H). .. Preparative chiral HPLC Instrument: Sepiatec: Prep SFC100; Column: Chiralpak IA 5μ 250 × 30 mm; eluent A: CO2; eluent B: 2-propanol + 0.4 vol % diethylamine; isocratic: 25% B; flow: 100 mL/min; temperature: 40° C.; BPR: 150 bar; UV: 280 nm Analytical chiral HPLC: Rt = 4.57 min Instrument: Agilent: 1260, Aurora SFC-module; column: Chiralpak IA 5μ 100 × 4.6 mm; eluent A: CO2; eluent B: 2-propanol + 0.4 vol % diethylamine, isocratic: 25% B; flow: 4 mL/min; temperature: 37.5° C.; BPR: 100 bar; UV: 280 nm 74 Intermediate 92; rac-2- bromo- propanamide 1H-NMR (400 MHz, DMSO-d6): δ ppm = 1.18 (d, J = 7.35 Hz, 3 H), 5.05 (q, J = 7.18 Hz, 1 H), 7.28 (s, 1 H), 7.32 (td, J = 8.30, 2.15 Hz, 1 H), 7.37-7.46 (m, 4 H), 7.47-7.56 (m, 4 H), 7.60 (s, 1 H), 7.85-8.35 (m, 2 H). ..
Article Title: Substituted aminothiazoles as DGKzeta inhibitors for immune activation
Article Snippet: Preparative chiral HPLC Instrument: Sepiatec: Prep SFC100; Column: Chiralpak IA 5μ 250 × 30mm; eluent A: CO2; eluent B: 2-propanol + 0.4 vol % diethylamine; isocratic: 30% B; flow: 100 mL/min; temperature: 40° C.; BPR: 150 bar; UV: 254 nm Analytical chiral HPLC: Rt = 1.72 min Instrument: Agilent: 1260, Aurora SFC-Modul; Column: Chiralpak IA 5μ 100 × 4.6mm; eluent A: CO2; eluent B: 2-propanol + 0.4 vol % diethylamine; isocratic: 30% B; flow: 4 mL/min; temperature: 37.5° C.; BPR: 100 bar; UV: 254 nm 62.2 Example 62 1H-NMR (400 MHz, DMSO-d6): δ ppm = 1.16 (d, J = 7.60 Hz, 3 H), 3.75 (s, 3 H), 5.06 (q, J = 6.76 Hz, 1 H), 6.93 (d, J = 8.87 Hz, 2 H), 7.22-7.27 (m, 1 H), 7.34 (t, J = 8.87 Hz, 2 H), 7.48 (d, J = 8.87 Hz, 2 H), 7.57 (s, 1 H), 7.64 (dd, J = 8.87, 5.07 Hz, 2 H), 7.79- 8.38 (m, 2 H). .. Preparative chiral HPLC Instrument: Sepiatec: Prep SFC100; Column: Chiralpak IA 5μ 250 × 30mm; eluent A: CO2; eluent B: 2-propanol + 0.4 vol % diethylamine; isocratic: 30% B; flow: 100 mL/min; temperature: 40° C.; BPR: 150 bar; UV: 254 nm Analytical chiral HPLC: Rt = 1.72 min Instrument: Agilent: 1260, Aurora SFC-Modul; Column: Chiralpak IA 5μ 100 × 4.6mm; eluent A: CO2; eluent B: 2-propanol + 0.4 vol % diethylamine; isocratic: 30% B; flow: 4 mL/min; temperature: 37.5° C.; BPR: 100 bar; UV: 254 nm 62.2 Example 62 1H-NMR (400 MHz, DMSO-d6): δ ppm = 1.16 (d, J = 7.60 Hz, 3 H), 3.75 (s, 3 H), 5.06 (q, J = 6.76 Hz, 1 H), 6.93 (d, J = 8.87 Hz, 2 H), 7.22-7.27 (m, 1 H), 7.34 (t, J = 8.87 Hz, 2 H), 7.48 (d, J = 8.87 Hz, 2 H), 7.57 (s, 1 H), 7.64 (dd, J = 8.87, 5.07 Hz, 2 H), 7.79- 8.38 (m, 2 H). .. Preparative chiral HPLC Instrument: Sepiatec: Prep SFC100; Column: Chiralpak IA 5μ 250 × 30mm; eluent A: CO2; eluent B: 2-propanol + 0.4 vol % diethylamine; isocratic: 30% B; flow: 100 mL/min; temperature: 40° C.; BPR: 150 bar; UV: 254 nm Analytical chiral HPLC: Rt = 2.61 min Instrument: Agilent: 1260, Aurora SFC-Modul; Column: Chiralpak IA 5μ 100 × 4.6mm; eluent A: CO2; eluent B: 2-propanol + 0.4 vol % diethylamine; isocratic: 30% B; flow: 4 mL/min; temperature: 37.5° C.; BPR: 100 bar; UV: 254 nm 62.2 Example 62.2 was determined to be (R)-2-(N-[4- amino-5-(4-methoxybenzoyl)thiazol-2-yl]-4- fluoro-anilino)propanamide, by means of X-ray crystal structure analysis.
Article Title: Substituted aminothiazoles as DGKzeta inhibitors for immune activation
Article Snippet: Preparative chiral HPLC Instrument: PrepCon Labomatic HPLC-3; Column: YMC Amylose SA 10μ, 250 × 50; eluent A: methyl tert-butyl ether + 0.1 vol % diethylamine; eluent B: methanol; isocratic: 50% A + 50% B; flow: 100 mL/min; temperature: 25° C.; UV: 254 nm Analytical chiral HPLC: Rt = 1.28 min Instrument: Thermo Fisher UltiMate 3000; Column: YMC Amylose SA 3μ, 100 × 4.6; eluent A: methyl tert-butyl ether + 0.1 vol % diethylamine; eluent B: acetonitrile; isocratic: 50% A + 50% B; flow: 1.4 mL/min; temperature: 25° C.; UV: 254 nm 174.2 Example 174 1H NMR (400 MHz, DMSO-d6) δ ppm = 1.15 (d, J = 7.35 Hz, 3 H), 3.73 (s, 3H), 5.06 (q, J = 7.35 Hz, 1 H), 7.01 (d, J = 9.13 Hz, 2 H), 7.16 (m, 2 H), 7.21 (s, 1 H), 7.27 (t, J = 72 Hz, 1 H), 7.46 (m, 2 H), 7.54 (m, 3 H), 8.15 (m, 2 H) LC-MS (method 2) Rt = 1.08 min MS (ESIpos): m/z = 463.2 [M + H]+ 31% yield 2-(N-[4-amino-5-[4- (difluoromethoxy)benzoyl] thiazol-2-yl]-4-methoxy- anilino)propanamide (enantiomer 2) Chiral HPLC Example 174.2 HPLC separation of rac-2-(N-[4-amino-5-[4-(difluoromethoxy)benzoyl]thiazol-2-yl]-4-methoxy- anilino)propanamide(316 mg, 0.66 mmol, Example 174 on a chiral column gave 121 mg (31% yield) of 2-(N-[4-amino-5-[4-(difluoromethoxy)benzoyl]thiazol-2-yl]-4-methoxy- anilino)propanamide, enantiomer 2. .. Preparative chiral HPLC Instrument: PrepCon Labomatic HPLC-3; Column: YMC Amylose SA 10μ, 250 × 50; eluent A: methyl tert-butyl ether + 0.1 vol % diethylamine; eluent B: methanol; isocratic: 50% A + 50% B; flow: 100 mL/min; temperature: 25° C.; UV: 254 nm Analytical chiral HPLC: Rt = 1.62 min Instrument: Thermo Fisher UltiMate 3000; Column: YMC Amylose SA 3μ, 100 × 4.6; eluent A: methyl tert-butyl ether + 0.1 vol % diethylamine; eluent B: acetonitrile; isocratic: 50% A + 50% B; flow: 1.4 mL/min; temperature: 25° C.; UV: 254 nm 175 Intermediate 116, rac-2- bromopropanamide 1H NMR (400 MHz, DMSO-d6) δ ppm = 1.16 (d, J = 7.35 Hz, 3 H), 5.07 (q, J = 7.35 Hz, 1 H), 7.26 (br s, 1 H), 7.33 (m, 3 H), 7.59 (br s, 1 H), 7.63 (m, 3 H), 7.71 (m, 2 H), 8.23 (m, 3 H), 8.38 (m, 1 H) Biotage (method X) LC-MS (method 2) Rt = 1.05 min MS (ESIpos): m/z = 430.1 [M + H]+ 66% yield rac-2-(N-[4-amino-5-(4- nitrobenzoyl)thiazol-2-yl]- 4-fluoro- anilino)propanamide 176 Intermediate 117, rac-2- bromopropanamide 1H NMR (500 MHz, DMSO-d6) δ ppm = 1.16 (d, J = 7.57 Hz, 3 H), 2.29 (s, 3 H), 3.80 (s, 3 H), 5.11 (q, J = 7.35 Hz, 1 H), 6.99 (d, J = 8.83 Hz, 2 H), 7.20 (s, 1 H), 7.36 (dd, J = 8.83 Hz, 2 H), 7.58 (m, 2 H), 7.61 (s, 1 H), 7.68 (m, 2 H) LC-MS (method 2) Rt = 1.16 min MS (ESIpos): m/z = 414.5 [M + H]+ 79% yield rac-2-(4-fluoro-N-[5-(4- methoxybenzoyl)-4- methyl-thiazol-2- yl]anilino)propanamide 176.1 (R)-2-(4-fluoro-N-[5-(4- and methoxybenzoyl)-4- 176.2 methyl-thiazol-2- yl]anilino)propanamide and (S)-2-(4-fluoro-N-[5- (4-methoxybenzoy)-4- methyl-thiazol-2- yl]anilino)propanamide 176.1 Example 176 1H NMR (500 MHz, DMSO-d6) δ ppm = 1.16 (d, J = 7.57 Hz, 3 H), 2.29 (s, 3 H), 3.80 (s, 3 H), 5.11 (q, J = 7.35 Hz, 1 H), 6.99 (d, J = 8.83 Hz, 2 H), 7.20 (s, 1 H), 7.36 (dd, J = 8.83 Hz, 2 H), 7.58 (m, 2 H), 7.61 (s, 1 H), 7.68 (m, 2 H) LC-MS (method 2) Rt = 1.16 min MS (ESIpos): m/z = 414.5 [M + H]+ 49% yield 2-(4-fluoro-N-[5-(4- methoxybenzoyl)-4- methyl-thiazol-2- yl]anilino)propanamide (enantiomer 1) Chiral HPLC Example 176.1 HPLC separation of rac-2-(4-fluoro-N-[5-(4-methoxybenzoyl)-4-methyl-thiazol-2- yl]anilino)propanamide(80 mg, 0.19 mmol, Example 176 on a chiral column gave 39 mg (49% yield) of 2-(4-fluoro-N-[5-(4-methoxybenzoyl)-4-methyl-thiazol-2-yl]anilino)propanamide, enantiomer 1. .. Preparative chiral HPLC Instrument: PrepCon Labomatic HPLC-4; Column: YMC Cellulose SC 10μ, 250 × 50; eluent A: hexane + 0.1 vol % diethylamine; eluent B: ethanol; isocratic: 80% A + 20% B; flow: 100 mL/min; temperature: 25° C.; UV: 254 nm Analytical chiral HPLC: Rt = 4.20 min Instrument: Thermo Fisher UltiMate 3000; Column: YMC Cellulose SC 3μ, 100 × 4.6; eluent A: hexane + 0.1 vol % diethylamine; eluent B: ethanol; isocratic: 80% A + 20% B; flow: 1.4 mL/min; temperature: 25° C.; UV: 254 nm 176.2 Example 176 1H NMR (500 MHz, DMSO-d6) δ ppm = 1.16 (d, J = 7.57 Hz, 3 H), 2.29 (s, 3 H), 3.80 (s, 3 H), 5.11 (q, J = 7.35 Hz, 1 H), 6.99 (d, J = 8.83 Hz, 2 H), 7.20 (s, 1 H), 7.36 (dd, J = 8.83 Hz, 2 H), 7.58 (m, 2 H), 7.61 (s, 1 H), 7.68 (m, 2 H) LC-MS (method 2) Rt = 1.16 min MS (ESIpos): m/z = 414.5 [M + H]+ 49% yield 2-(4-fluoro-N-[5-(4- methoxybenzoyl)-4- methyl-thiazol-2- yl]anilino)propanamide (enantiomer 2) Chiral HPLC Example 176.2 HPLC separation of rac-2-(4-fluoro-N-[5-(4-methoxybenzoyl)-4-methyl-thiazol-2- yl]anilino)propanamide(80 mg, 0.19 mmol, Example 176 on a chiral column gave 31 mg (39% yield) of 2-(4-fluoro-N-[5-(4-methoxybenzoyl)-4-methyl-thiazol-2-yl]anilino)propanamide, enantiomer 2.
Article Title: Substituted aminothiazoles as DGKzeta inhibitors for immune activation
Article Snippet: Preparative chiral HPLC Instrument: Sepiatec: Prep SFC100; Column: Chiralpak IA 5μ 250 × 30mm; eluent A: CO2; eluent B: 2-propanol + 0.4 vol % diethylamine; isocratic: 25% B; flow: 100 mL/min; temperature: 40° C.; BPR: 150 bar; UV: 280 nm Analytical chiral HPLC: Rt = 3.08 min Instrument: Agilent: 1260, Aurora SFC-Modul; column: Chiralpak IA 5μ 100 × 4.6mm; eluent A: CO2; eluent B: 2-Propanol + 0.4% diethylamine (99%); isocratic; 25%B; flow: 4 mL/min; temperature: 37.5° C.; BPR: 100 bar; UV: 280 nm 61.2 Example 61.2 was determined to be (R)-2-(N- (4-amino-5-benzoyl-thiazol-2-yl)-4-fluoro- anilino)propanamide, by means of X-ray crystal structure analysis. .. Preparative chiral HPLC Instrument: Sepiatec: Prep SFC100; Column: Chiralpak IA 5μ 250 × 30mm; eluent A: CO2; eluent B: 2-propanol + 0.4 vol % diethylamine; isocratic: 25% B; flow: 100 mL/min; temperature: 40° C.; BPR: 150 bar; UV: 280 nm Analytical chiral HPLC: Rt = 3.08 min Instrument: Agilent: 1260, Aurora SFC-Modul; column: Chiralpak IA 5μ 100 × 4.6mm; eluent A: CO2; eluent B: 2-Propanol + 0.4% diethylamine (99%); isocratic; 25%B; flow: 4 mL/min; temperature: 37.5° C.; BPR: 100 bar; UV: 280 nm 61.2 Example 61.2 was determined to be (R)-2-(N- (4-amino-5-benzoyl-thiazol-2-yl)-4-fluoro- anilino)propanamide, by means of X-ray crystal structure analysis. ..
Article Title: Substituted aminothiazoles as DGKzeta inhibitors for immune activation
Article Snippet: Preparative chiral HPLC Instrument: Sepiatec: Prep SFC100; Column: Chiralpak IB 5μ 250 × 30 mm; eluent A: CO2; eluent B: 2-propanol + 0.4 vol % diethylamine; isocratic: 35% B; flow: 100 mL/min; temperature: 40° C.; BPR: 150 bar; UV: 254 nm Analytical chiral HPLC: Rt = 4.42 min Instrument: Agilent: 1260, Aurora SFC-Modul; Column: Chiralpak IB 5μ 100 × 4.6 mm; eluent A: CO2; eluent B: 2-propanol + 0.4 vol % diethylamine; isocratic: 35% B; flow: 4 mL/min; temperature: 37.5° C.; BPR: 100 bar; UV: 254 nm 171 Intermediate 111, rac-2- bromopropanamide 1H NMR (400 MHz, DMSO-d6) δ ppm = 1.16 (d, J = 7.35 Hz, 3 H), 5.07 (q, J = 7.35 Hz, 1 H), 7.29 (m, 5 H), 7.58 (m, 1 H), 7.63 (m, 2 H), 7.76 (m, 2 H), 8.21 (m, 2 H) Biotage (method X) LC-MS (method 2) Rt = 1.21 min MS (ESIpos): m/z = 511.0 [M + H]+ 39% yield rac-2-(N-[4-amino-5-(4- iodobenzoyl)thiazol-2-yl] 4-fluoro- anilino)propanamide 172 Intermediate 113, rac-2- bromopropanamide 1H NMR (400 MHz, DMSO-d6) δ ppm = 1.20 (d, J = 7.35 Hz, 3 H), 3.87 (s, 3 H), 5.03 (q, J = 7.35 Hz, 1 H), 6.85 (dd, J = 8.62, 0.76 Hz, 1 H), 7.31 (s, 1 H), 7.52 (m, 1 H), 7.64 (s, 1 H), 7.76 (m, 2 H), 7.84 (dd, J = 8.49, 2.41 Hz, 1 H), 8.14 (m, 2 H), 8.37 (m, 1 H) Biotage (method Y) LC-MS (method 2) Rt = 1.14 min MS (ESIpos): m/z = 450.3 [M + H]+ 38% yield rac-2-(N-[4-amino-5-(6- methoxypyridine-3- carbonyl)thiazol-2-yl]-4- chloro-3-fluoro- anilino)propanamide 172.1 (R)-2-(N-[4-amino-5-(6- and methoxypyridine-3- 172.2 carbonyl)thiazol-2-yl]-4- chloro-3-fluoro- anilino)propanamide and (S)-2-(N-[4-amino-5-(6- methoxypyridine-3- carbonyl)thiazol-2-yl]-4- chloro-3-fluoro- anilino)propanamide 172.1 Example 172 1H NMR (400 MHz, DMSO-d6) δ ppm = 1.20 (d, J = 7.35 Hz, 3 H), 3.87 (s, 3 H), 5.03 (q, J = 7.35 Hz, 1 H), 6.85 (dd, J = 8.62, 0.76 Hz, 1 H), 7.31 (s, 1 H), 7.52 (m, 1 H), 7.64 (s, 1 H), 7.76 (m, 2 H), 7.84 (dd, J = 8.49, 2.41 Hz, 1 H), 8.14 (m, 2 H), 8.37 (m, 1 H) LC-MS (method 2) Rt = 1.14 min MS (ESIpos): m/z = 450.3 [M + H]+ 24% yield 2-(N-[4-amino-5-(6- methoxypyridine-3- carbonyl)thiazol-2-yl]-4- chloro-3-fluoro- anilino)propanamide (enantiomer 1) Chiral HPLC Example 172.1 HPLC separation of rac-2-(N-[4-amino-5-(6-methoxypyridine-3-carbonyl)thiazol-2-yl]-4-chloro-3- fluoro-anilino)propanamide(312 mg, 0.69 mmol, Example 172) on a chiral column gave 78 mg (24% yield) of 2-(N-[4-amino-5-(6-methoxypyridine-3-carbonyl)thiazol-2-yl]-4-chloro-3-fluoro- anilino)propanamide, enantiomer 1. .. Preparative chiral HPLC Instrument: Sepiatec: Prep SFC100; Column: Chiralpak IG 5μ 250 × 30 mm; eluent A: CO2; eluent B: methanol + 0.2 vol % aqueous ammonia (32%); isocratic: 30% B; flow: 100 mL/min; temperature: 40° C.; BPR: 150 bar; UV: 254 nm Analytical chiral HPLC: Rt = 2.61 min Instrument: Agilent: 1260, Aurora SFC-Modul; Column: Chiralpak IG 5μ 100 × 4.6 mm; eluent A: CO2; eluent B: methanol + 0.2 vol % aqueous ammonia (32%); isocratic: 30% B; flow: 4 mL/min; temperature: 37.5° C.; BPR: 100 bar; UV: 254 nm 172.2 Example 172 1H NMR (400 MHz, DMSO-d6) δ ppm = 1.20 (d, J = 7.35 Hz, 3 H), 3.87 (s, 3 H), 5.03 (q, J = 7.35 Hz, 1 H), 6.85 (dd, J = 8.62, 0.76 Hz, 1 H), 7.31 (s, 1 H), 7.52 (m, 1 H), 7.64 (s, 1 H), 7.76 (m, 2 H), 7.84 (dd, J = 8.49, 2.41 Hz, 1 H), 8.14 (m, 2 H), 8.37 (m, 1 H) LC-MS (method 2) Rt = 1.14 min MS (ESIpos): m/z = 450.3 [M + H]+ 23% yield 2-(N-[4-amino-5-(6- methoxypyridine-3- carbonyl)thiazol-2-yl]-4- chloro-3-fluoro- anilino)propanamide (enantiomer 2) Chiral HPLC Example 172.2 HPLC separation of rac-2-(N-[4-amino-5-(6-methoxypyridine-3-carbonyl)thiazol-2-yl]-4-chloro-3- fluoro-anilino)propanamide(312 mg, 0.69 mmol, Example 172) on a chiral column gave 73 mg (23% yield) of 2-(N-[4-amino-5-(6-methoxypyridine-3-carbonyl)thiazol-2-yl]-4-chloro-3-fluoro- anilino)propanamide, enantiomer 2. .. Preparative chiral HPLC Instrument: Sepiatec: Prep SFC100; Column: Chiralpak IG 5μ 250 × 30 mm; eluent A: CO2; eluent B: methanol + 0.2 vol % aqueous ammonia (32%); isocratic: 30% B; flow: 100 mL/min; temperature: 40° C.; BPR: 150 bar UV: 254 nm Analytical chiral HPLC: Rt = 3.40 min Instrument: Agilent: 1260, Aurora SFC-Modul; Column: Chiralpak IG 5μ 100 × 4.6 mm; eluent A: CO2; eluent B: methanol + 0.2 vol % aqueous ammonia (32%); isocratic: 30% B; flow: 4 mL/min; temperature: 37.5° C.; BPR: 100 bar; UV: 254 nm 173 Intermediate 114, rac-2- bromopropanamide 1H NMR (400 MHz, DMSO-d6) δ ppm = 1.16 (d, J = 7.35 Hz, 3 H), 5.07 (m, 1 H), 6.94 (m, 2 H), 7.06 (m, 2 H), 7.20 (m, 1 H), 7.25 (s, 1 H), 7.33 (dd, J = 8.87 Hz, 2 H), 7.41 (m, 2 H), 7.53 (m, 2 H), 7.58 (s, 1 H), 7.64 (m, 2 H), 8.14 (m, 2 H) Biotage (method X) LC-MS (method 2) Rt = 1.26 min MS (ESIpos): m/z = 477.2 [M + H]+ 56% yield rac-2-(N-[4-amino-5-(4- phenoxybenzoyl)thiazol-2- yl]-4-fluoro- anilino)propanamide 174 Intermediate 115, rac-2- bromopropanamide 1H NMR (400 MHz, DMSO-d6) δ ppm = 1.15 (d, J = 7.35 Hz, 3 H), 3.73 (s, 3H), 5.06 (q, J = 7.35 Hz, 1 H), 7.01 (d, J = 9.13 Hz, 2 H), 7.16 (m, 2 H), 7.21 (s, 1 H), 7.27 (t, J = 72 Hz, 1 H), 7.46 (m, 2 H), 7.54 (m, 3 H), 8.15 (m, 2 H) Biotage (method X) LC-MS (method 2) Rt = 1.08 min MS (ESIpos): m/z = 463.2 [M + H]+ 77% yield rac-2-(N-[4-amino-5-[4- (difluoromethoxy)benzoyl] thiazol-2-yl]-4-methoxy- anilino)propanamide 174.1 (R)-2-(N-[4-amino-5-[4- and (difluoromethoxy)benzoyl] 174.2 thiazol-2-yl]-4-methoxy- anilino)propanamide and (S)-2-(N-[4-amino-5-[4- (difluoromethoxy)benzoyl] thiazol-2-yl]-4-methoxy- anilino)propanamide 174.1 Example 174 1H NMR (400 MHz, DMSO-d6) δ ppm = 1.15 (d, J = 7.35 Hz, 3 H), 3.73 (s, 3H), 5.06 (q, J = 7.35 Hz, 1 H), 7.01 (d, J = 9.13 Hz, 2 H), 7.16 (m, 2 H), 7.21 (s, 1 H), 7.27 (t, J = 72 Hz, 1 H), 7.46 (m, 2 H), 7.54 (m, 3 H), 8.15 (m, 2 H) LC-MS (method 2) Rt = 1.08 min MS (ESIpos): m/z = 463.2 [M + H]+ 31% yield 2-(N-[4-amino-5-[4- (difluoromethoxy)benzoyl] thiazol-2-yl]-4-methoxy- anilino)propanamide (enantiomer 1) Chiral HPLC Example 174.1 HPLC separation of rac-2-(N-[4-amino-5-[4-(difluoromethoxy)benzoyl]thiazol-2-yl]-4-methoxy- anilino)propanamide(316 mg, 0.66 mmol, Example 174 on a chiral column gave 121 mg (31% yield) of 2-(N-[4-amino-5-[4-(difluoromethoxy)benzoyl]thiazol-2-yl]-4-methoxy- anilino)propanamide, enantiomer 1.
Article Title: Substituted aminothiazoles as DGKzeta inhibitors for immune activation
Article Snippet: Preparative chiral HPLC Instrument: Sepiatec: Prep SFC100; Column: Chiralpak IG 5μ 250 × 30 mm; eluent A: CO2; eluent B: ethanol + 0.2 vol % aqueous ammonia (32%); isocratic: 15% B; flow: 100 mL/min; temperature: 40° C.; BPR: 150 bar; UV: 254 nm Analytical chiral HPLC: Rt = 2.81 min Instrument: Agilent: 1260, Aurora SFC-Modul; Column: Chiralpak IG 5μ 100 × 4.6 mm; eluent A: CO2; eluent B: ethanol + 0.2 vol % aqueous ammonia (32%); isocratic: 25% B; flow: 4 mL/min; temperature: 37.5° C.; BPR: 100 bar; UV: 254 nm 187.2 Example 187 1H NMR (400 MHz, DMSO-d6) δ ppm = 1.15 (d, J = 7.60 Hz, 3 H), 2.45 (s, 3 H), 5.06 (br s, 1 H), 7.10-7.23 (m, 1 H), 7.23-7.28 (m, 2 H), 7.33 (dd, J = 8.74 Hz, 2 H), 7.59 (br s, 1 H), 7.63 (dd, J = 8.87, 5.07 Hz, 2 H), 8.05-8.42 (m, 2 H), 8.46 (d, J = 5.07 Hz, 1 H) 23% yield 2-(N-[4-amino-5-(2- methylpyridine-4- carbonyl)thiazol-2-yl]-4- fluoro-anilino)propanamide (enantiomer 2) Chiral HPLC Example 187.2 HPLC separation of rac-2-(N-[4-amino-5-(2-methylpyridine-4-carbonyl)thiazol-2-yl]-4-fluoro- anilino)propanamide(333 mg, 0.83 mmol, Example 187) on a chiral column gave 78 mg (23% yield) of 2-(N-[4-amino-5-(2-methylpyridine-4-carbonyl)thiazol-2-yl]-4-fluoro-anilino)propanamide, enantiomer 2. .. Preparative chiral HPLC Instrument: Sepiatec: Prep SFC100; Column: Chiralpak IG 5μ 250 × 30 mm; eluent A: CO2; eluent B: ethanol + 0.2 vol % aqueous ammonia (32%); isocratic: 15% B; flow: 100 mL/min; temperature: 40° C.; BPR: 150 bar; UV: 254 nm Analytical chiral HPLC: Rt = 3.56 min Instrument: Agilent: 1260, Aurora SFC-Modul; Column: Chiralpak IG 5μ 100 × 4.6 mm; eluent A: CO2; eluent B: ethanol + 0.2 vol % aqueous ammonia (32%); isocratic: 25% B; flow: 4 mL/min; temperature: 37.5° C.; BPR: 100 bar; UV: 254 nm 188 Intermediate 129, rac-2- bromopropanamide 1H NMR (400 MHz, DMSO-d6) δ ppm = 1.16 (d, J = 7.35 Hz, 3 H), 4.99-5.13 (m, 1 H), 6.97 (t, J = 52 Hz, 1 H), 7.27 (s, 1 H), 7.33 (t J = 8.87 Hz, 2 H), 7.56-7.61 (m, 2 H), 7.64 (dd, J = 8.87, 5.07 Hz, 2 H), 7.67 (s, 1 H), 8.20-8.57 (m, 2 H), 8.73 (d, J = 4.56 Hz, 1 H) RP-HPLC (method C basic) LC-MS (method 2) Rt = 1.00 min MS (ESIpos): m/z = 436.3 [M + H]+ 13% yield rac-2-(N-[4-amino-5-[2- (difluoromethyl)pyridine-4- carbonyl]thiazol-2-yl]-4- fluoro-anilino)propanamide 189 Intermediate 130, rac-2- bromopropanamide 1H NMR (400 MHz, DMSO-d6) δ ppm = 1.22 (d, J = 7.35 Hz, 3 H), 5.05 (q, J = 7.35 Hz, 1 H), 5.74 (s, 1 H), 7.32 (s, 1 H), 7.37-7.44 (m, 2 H), 7.48-7.55 (m, 2 H), 7.56-7.65 (m, 3 H), 8.06-8.42 (m, 2 H), 8.68-8.73 (m, 2 H) RP-HPLC (method C basic) LC-MS (method 2) Rt = 0.80 min MS (ESIpos): m/z = 368.2 [M + H]+ 17% yield rac-2-[(4-amino-5-benzoyl- thiazol-2-yl)-(4- pyridyl)amino]propanamide 190 Intermediate 131, rac-2- bromopropanamide 1H NMR (400 MHz, DMSO-d6) δ ppm = 1.15 (d, J = 7.35 Hz, 3 H), 3.83 (s, 3H), 5.00-5.11 (m, 1 H), 6.76 (s, 1 H), 6.99 (dd, J = 5.20, 1.39 Hz, 1 H), 7.26 (s, 1 H), 7.30- 7.37 (m, 2 H), 7.59 (s, 1 H), 7.63 (dd, J = 8.87, 5.07 Hz, 2 H), 8.17-8.20 (m, 1 H), 8.21-8.48 (m, 2 H) RP-HPLC (method C basic) LC-MS (method 2) Rt = 0.98 min MS (ESIpos): m/z = 416.2 [M + H]+ 29% yield rac-2-(N-[4-amino-5-(2- methoxypyridine-4- carbonyl)thiazol-2-yl]-4- fluoro-anilino)propanamide 191 Intermediate 133; rac-2- bromopropanamide 1H NMR (400 MHz, DMSO-d6) δ ppm = 8.12 (br s, 2H), 7.57 (s, 1H), 7.46-7.53 (m, 3H), 7.34-7.44 (m, 4H), 7.21-7.28 (m, 2H), 4.98- 5.10 (m, 1H), 3.87 (s, 3H), 1.16 (d, J = 7.4 Hz, 3H). ..
Article Title: Substituted aminothiazoles as DGKzeta inhibitors for immune activation
Article Snippet: Preparative chiral HPLC Instrument: Sepiatec: Prep SFC100; Column: Chiralpak IC 5μ 250 × 30mm; eluent A: CO2; eluent B: methanol + 0.2 vol % aqueous ammonia (32%); isocratic: 40% B; flow: 100 mL/min; temperature: 40° C.; BPR: 150 bar; UV: 254 nm Analytical chiral HPLC: Rt = 2.66 min Instrument: Agilent: 1260, Aurora SFC-Modul; Column: Chiralpak IC 5μ 100 × 4.6 mm; eluent A: CO2; eluent B: methanol + 0.2 vol % aqueous ammonia (32%); isocratic: 40% B; flow: 4 mL/min; temperature: 37.5° C.; BPR: 100 bar; UV: 254 nm 79 rac-2-[(4-amino-5-benzoyl- thiazol-2-yl)-(3- Intermediate 95; rac-2- bromo- propanamide 1H-NMR (400 MHz, DMSO-d6): δ ppm = 1.17 (d, J =7.35 Hz, 3 H), 5.07 (q, J =7.10 Hz, 1 H), 5.76 (s, 1 H), 7.30 (s, 1 H), 7.35- 7.43 (m, 3 H), 7.47-7.51 (m, 2 H), 7.54 (dd, J = 8.1 1, 4.82 Hz, 1 H), 7.64 (s, 1 H), 7.90-8.44 (m, 2 H), 8.04-8.08 (m, 1 H), 8.63 (dd, J = 4.56, 1.52 Hz, 1 H), 8.74 (d, J = 2.03 Hz, 1 H). .. Preparative chiral HPLC Instrument: Sepiatec: Prep SFC100; Column: Chiralpak IC 5μ 250 × 30mm; eluent A: CO2; eluent B: methanol + 0.2 vol % aqueous ammonia (32%); isocratic: 40% B; flow: 100 mL/min; temperature: 40° C.; BPR: 150 bar; UV: 254 nm Analytical chiral HPLC: Rt = 2.66 min Instrument: Agilent: 1260, Aurora SFC-Modul; Column: Chiralpak IC 5μ 100 × 4.6 mm; eluent A: CO2; eluent B: methanol + 0.2 vol % aqueous ammonia (32%); isocratic: 40% B; flow: 4 mL/min; temperature: 37.5° C.; BPR: 100 bar; UV: 254 nm 79 rac-2-[(4-amino-5-benzoyl- thiazol-2-yl)-(3- Intermediate 95; rac-2- bromo- propanamide 1H-NMR (400 MHz, DMSO-d6): δ ppm = 1.17 (d, J =7.35 Hz, 3 H), 5.07 (q, J =7.10 Hz, 1 H), 5.76 (s, 1 H), 7.30 (s, 1 H), 7.35- 7.43 (m, 3 H), 7.47-7.51 (m, 2 H), 7.54 (dd, J = 8.1 1, 4.82 Hz, 1 H), 7.64 (s, 1 H), 7.90-8.44 (m, 2 H), 8.04-8.08 (m, 1 H), 8.63 (dd, J = 4.56, 1.52 Hz, 1 H), 8.74 (d, J = 2.03 Hz, 1 H). ..
Article Title: Substituted aminothiazoles as DGKzeta inhibitors for immune activation
Article Snippet: Preparative chiral HPLC Instrument: Sepiatec: Prep SFC100; Column: Chiralpak IG 5μ 250 × 30 mm; eluent A: CO2; eluent B: methanol + 0.2 vol % aqueous ammonia (32%); isocratic: 30% B; flow: 100 mL/min; temperature: 40° C.; BPR: 150 bar; UV: 254 nm Analytical chiral HPLC: Rt = 2.61 min Instrument: Agilent: 1260, Aurora SFC-Modul; Column: Chiralpak IG 5μ 100 × 4.6 mm; eluent A: CO2; eluent B: methanol + 0.2 vol % aqueous ammonia (32%); isocratic: 30% B; flow: 4 mL/min; temperature: 37.5° C.; BPR: 100 bar; UV: 254 nm 172.2 Example 172 1H NMR (400 MHz, DMSO-d6) δ ppm = 1.20 (d, J = 7.35 Hz, 3 H), 3.87 (s, 3 H), 5.03 (q, J = 7.35 Hz, 1 H), 6.85 (dd, J = 8.62, 0.76 Hz, 1 H), 7.31 (s, 1 H), 7.52 (m, 1 H), 7.64 (s, 1 H), 7.76 (m, 2 H), 7.84 (dd, J = 8.49, 2.41 Hz, 1 H), 8.14 (m, 2 H), 8.37 (m, 1 H) LC-MS (method 2) Rt = 1.14 min MS (ESIpos): m/z = 450.3 [M + H]+ 23% yield 2-(N-[4-amino-5-(6- methoxypyridine-3- carbonyl)thiazol-2-yl]-4- chloro-3-fluoro- anilino)propanamide (enantiomer 2) Chiral HPLC Example 172.2 HPLC separation of rac-2-(N-[4-amino-5-(6-methoxypyridine-3-carbonyl)thiazol-2-yl]-4-chloro-3- fluoro-anilino)propanamide(312 mg, 0.69 mmol, Example 172) on a chiral column gave 73 mg (23% yield) of 2-(N-[4-amino-5-(6-methoxypyridine-3-carbonyl)thiazol-2-yl]-4-chloro-3-fluoro- anilino)propanamide, enantiomer 2. .. Preparative chiral HPLC Instrument: Sepiatec: Prep SFC100; Column: Chiralpak IG 5μ 250 × 30 mm; eluent A: CO2; eluent B: methanol + 0.2 vol % aqueous ammonia (32%); isocratic: 30% B; flow: 100 mL/min; temperature: 40° C.; BPR: 150 bar UV: 254 nm Analytical chiral HPLC: Rt = 3.40 min Instrument: Agilent: 1260, Aurora SFC-Modul; Column: Chiralpak IG 5μ 100 × 4.6 mm; eluent A: CO2; eluent B: methanol + 0.2 vol % aqueous ammonia (32%); isocratic: 30% B; flow: 4 mL/min; temperature: 37.5° C.; BPR: 100 bar; UV: 254 nm 173 Intermediate 114, rac-2- bromopropanamide 1H NMR (400 MHz, DMSO-d6) δ ppm = 1.16 (d, J = 7.35 Hz, 3 H), 5.07 (m, 1 H), 6.94 (m, 2 H), 7.06 (m, 2 H), 7.20 (m, 1 H), 7.25 (s, 1 H), 7.33 (dd, J = 8.87 Hz, 2 H), 7.41 (m, 2 H), 7.53 (m, 2 H), 7.58 (s, 1 H), 7.64 (m, 2 H), 8.14 (m, 2 H) Biotage (method X) LC-MS (method 2) Rt = 1.26 min MS (ESIpos): m/z = 477.2 [M + H]+ 56% yield rac-2-(N-[4-amino-5-(4- phenoxybenzoyl)thiazol-2- yl]-4-fluoro- anilino)propanamide 174 Intermediate 115, rac-2- bromopropanamide 1H NMR (400 MHz, DMSO-d6) δ ppm = 1.15 (d, J = 7.35 Hz, 3 H), 3.73 (s, 3H), 5.06 (q, J = 7.35 Hz, 1 H), 7.01 (d, J = 9.13 Hz, 2 H), 7.16 (m, 2 H), 7.21 (s, 1 H), 7.27 (t, J = 72 Hz, 1 H), 7.46 (m, 2 H), 7.54 (m, 3 H), 8.15 (m, 2 H) Biotage (method X) LC-MS (method 2) Rt = 1.08 min MS (ESIpos): m/z = 463.2 [M + H]+ 77% yield rac-2-(N-[4-amino-5-[4- (difluoromethoxy)benzoyl] thiazol-2-yl]-4-methoxy- anilino)propanamide 174.1 (R)-2-(N-[4-amino-5-[4- and (difluoromethoxy)benzoyl] 174.2 thiazol-2-yl]-4-methoxy- anilino)propanamide and (S)-2-(N-[4-amino-5-[4- (difluoromethoxy)benzoyl] thiazol-2-yl]-4-methoxy- anilino)propanamide 174.1 Example 174 1H NMR (400 MHz, DMSO-d6) δ ppm = 1.15 (d, J = 7.35 Hz, 3 H), 3.73 (s, 3H), 5.06 (q, J = 7.35 Hz, 1 H), 7.01 (d, J = 9.13 Hz, 2 H), 7.16 (m, 2 H), 7.21 (s, 1 H), 7.27 (t, J = 72 Hz, 1 H), 7.46 (m, 2 H), 7.54 (m, 3 H), 8.15 (m, 2 H) LC-MS (method 2) Rt = 1.08 min MS (ESIpos): m/z = 463.2 [M + H]+ 31% yield 2-(N-[4-amino-5-[4- (difluoromethoxy)benzoyl] thiazol-2-yl]-4-methoxy- anilino)propanamide (enantiomer 1) Chiral HPLC Example 174.1 HPLC separation of rac-2-(N-[4-amino-5-[4-(difluoromethoxy)benzoyl]thiazol-2-yl]-4-methoxy- anilino)propanamide(316 mg, 0.66 mmol, Example 174 on a chiral column gave 121 mg (31% yield) of 2-(N-[4-amino-5-[4-(difluoromethoxy)benzoyl]thiazol-2-yl]-4-methoxy- anilino)propanamide, enantiomer 1. .. Preparative chiral HPLC Instrument: PrepCon Labomatic HPLC-3; Column: YMC Amylose SA 10μ, 250 × 50; eluent A: methyl tert-butyl ether + 0.1 vol % diethylamine; eluent B: methanol; isocratic: 50% A + 50% B; flow: 100 mL/min; temperature: 25° C.; UV: 254 nm Analytical chiral HPLC: Rt = 1.28 min Instrument: Thermo Fisher UltiMate 3000; Column: YMC Amylose SA 3μ, 100 × 4.6; eluent A: methyl tert-butyl ether + 0.1 vol % diethylamine; eluent B: acetonitrile; isocratic: 50% A + 50% B; flow: 1.4 mL/min; temperature: 25° C.; UV: 254 nm 174.2 Example 174 1H NMR (400 MHz, DMSO-d6) δ ppm = 1.15 (d, J = 7.35 Hz, 3 H), 3.73 (s, 3H), 5.06 (q, J = 7.35 Hz, 1 H), 7.01 (d, J = 9.13 Hz, 2 H), 7.16 (m, 2 H), 7.21 (s, 1 H), 7.27 (t, J = 72 Hz, 1 H), 7.46 (m, 2 H), 7.54 (m, 3 H), 8.15 (m, 2 H) LC-MS (method 2) Rt = 1.08 min MS (ESIpos): m/z = 463.2 [M + H]+ 31% yield 2-(N-[4-amino-5-[4- (difluoromethoxy)benzoyl] thiazol-2-yl]-4-methoxy- anilino)propanamide (enantiomer 2) Chiral HPLC Example 174.2 HPLC separation of rac-2-(N-[4-amino-5-[4-(difluoromethoxy)benzoyl]thiazol-2-yl]-4-methoxy- anilino)propanamide(316 mg, 0.66 mmol, Example 174 on a chiral column gave 121 mg (31% yield) of 2-(N-[4-amino-5-[4-(difluoromethoxy)benzoyl]thiazol-2-yl]-4-methoxy- anilino)propanamide, enantiomer 2.
Nuclear Magnetic Resonance:Article Title: Substituted aminothiazoles as DGKzeta inhibitors for immune activation
Article Snippet: Preparative chiral HPLC Instrument: PrepCon Labomatic HPLC-3; Column: YMC Amylose SA 10μ, 250 × 50; eluent A: methyl tert-butyl ether + 0.1 vol % diethylamine; eluent B: methanol; isocratic: 50% A + 50% B; flow: 100 mL/min; temperature: 25° C.; UV: 254 nm Analytical chiral HPLC: Rt = 1.28 min Instrument: Thermo Fisher UltiMate 3000; Column: YMC Amylose SA 3μ, 100 × 4.6; eluent A: methyl tert-butyl ether + 0.1 vol % diethylamine; eluent B: acetonitrile; isocratic: 50% A + 50% B; flow: 1.4 mL/min; temperature: 25° C.; UV: 254 nm 174.2 Example 174 1H NMR (400 MHz, DMSO-d6) δ ppm = 1.15 (d, J = 7.35 Hz, 3 H), 3.73 (s, 3H), 5.06 (q, J = 7.35 Hz, 1 H), 7.01 (d, J = 9.13 Hz, 2 H), 7.16 (m, 2 H), 7.21 (s, 1 H), 7.27 (t, J = 72 Hz, 1 H), 7.46 (m, 2 H), 7.54 (m, 3 H), 8.15 (m, 2 H) LC-MS (method 2) Rt = 1.08 min MS (ESIpos): m/z = 463.2 [M + H]+ 31% yield 2-(N-[4-amino-5-[4- (difluoromethoxy)benzoyl] thiazol-2-yl]-4-methoxy- anilino)propanamide (enantiomer 2) Chiral HPLC Example 174.2 HPLC separation of rac-2-(N-[4-amino-5-[4-(difluoromethoxy)benzoyl]thiazol-2-yl]-4-methoxy- anilino)propanamide(316 mg, 0.66 mmol, Example 174 on a chiral column gave 121 mg (31% yield) of 2-(N-[4-amino-5-[4-(difluoromethoxy)benzoyl]thiazol-2-yl]-4-methoxy- anilino)propanamide, enantiomer 2. .. Preparative chiral HPLC Instrument: PrepCon Labomatic HPLC-3; Column: YMC Amylose SA 10μ, 250 × 50; eluent A: methyl tert-butyl ether + 0.1 vol % diethylamine; eluent B: methanol; isocratic: 50% A + 50% B; flow: 100 mL/min; temperature: 25° C.; UV: 254 nm Analytical chiral HPLC: Rt = 1.62 min Instrument: Thermo Fisher UltiMate 3000; Column: YMC Amylose SA 3μ, 100 × 4.6; eluent A: methyl tert-butyl ether + 0.1 vol % diethylamine; eluent B: acetonitrile; isocratic: 50% A + 50% B; flow: 1.4 mL/min; temperature: 25° C.; UV: 254 nm 175 Intermediate 116, rac-2- bromopropanamide 1H NMR (400 MHz, DMSO-d6) δ ppm = 1.16 (d, J = 7.35 Hz, 3 H), 5.07 (q, J = 7.35 Hz, 1 H), 7.26 (br s, 1 H), 7.33 (m, 3 H), 7.59 (br s, 1 H), 7.63 (m, 3 H), 7.71 (m, 2 H), 8.23 (m, 3 H), 8.38 (m, 1 H) Biotage (method X) LC-MS (method 2) Rt = 1.05 min MS (ESIpos): m/z = 430.1 [M + H]+ 66% yield rac-2-(N-[4-amino-5-(4- nitrobenzoyl)thiazol-2-yl]- 4-fluoro- anilino)propanamide 176 Intermediate 117, rac-2- bromopropanamide 1H NMR (500 MHz, DMSO-d6) δ ppm = 1.16 (d, J = 7.57 Hz, 3 H), 2.29 (s, 3 H), 3.80 (s, 3 H), 5.11 (q, J = 7.35 Hz, 1 H), 6.99 (d, J = 8.83 Hz, 2 H), 7.20 (s, 1 H), 7.36 (dd, J = 8.83 Hz, 2 H), 7.58 (m, 2 H), 7.61 (s, 1 H), 7.68 (m, 2 H) LC-MS (method 2) Rt = 1.16 min MS (ESIpos): m/z = 414.5 [M + H]+ 79% yield rac-2-(4-fluoro-N-[5-(4- methoxybenzoyl)-4- methyl-thiazol-2- yl]anilino)propanamide 176.1 (R)-2-(4-fluoro-N-[5-(4- and methoxybenzoyl)-4- 176.2 methyl-thiazol-2- yl]anilino)propanamide and (S)-2-(4-fluoro-N-[5- (4-methoxybenzoy)-4- methyl-thiazol-2- yl]anilino)propanamide 176.1 Example 176 1H NMR (500 MHz, DMSO-d6) δ ppm = 1.16 (d, J = 7.57 Hz, 3 H), 2.29 (s, 3 H), 3.80 (s, 3 H), 5.11 (q, J = 7.35 Hz, 1 H), 6.99 (d, J = 8.83 Hz, 2 H), 7.20 (s, 1 H), 7.36 (dd, J = 8.83 Hz, 2 H), 7.58 (m, 2 H), 7.61 (s, 1 H), 7.68 (m, 2 H) LC-MS (method 2) Rt = 1.16 min MS (ESIpos): m/z = 414.5 [M + H]+ 49% yield 2-(4-fluoro-N-[5-(4- methoxybenzoyl)-4- methyl-thiazol-2- yl]anilino)propanamide (enantiomer 1) Chiral HPLC Example 176.1 HPLC separation of rac-2-(4-fluoro-N-[5-(4-methoxybenzoyl)-4-methyl-thiazol-2- yl]anilino)propanamide(80 mg, 0.19 mmol, Example 176 on a chiral column gave 39 mg (49% yield) of 2-(4-fluoro-N-[5-(4-methoxybenzoyl)-4-methyl-thiazol-2-yl]anilino)propanamide, enantiomer 1. .. Preparative chiral HPLC Instrument: PrepCon Labomatic HPLC-4; Column: YMC Cellulose SC 10μ, 250 × 50; eluent A: hexane + 0.1 vol % diethylamine; eluent B: ethanol; isocratic: 80% A + 20% B; flow: 100 mL/min; temperature: 25° C.; UV: 254 nm Analytical chiral HPLC: Rt = 4.20 min Instrument: Thermo Fisher UltiMate 3000; Column: YMC Cellulose SC 3μ, 100 × 4.6; eluent A: hexane + 0.1 vol % diethylamine; eluent B: ethanol; isocratic: 80% A + 20% B; flow: 1.4 mL/min; temperature: 25° C.; UV: 254 nm 176.2 Example 176 1H NMR (500 MHz, DMSO-d6) δ ppm = 1.16 (d, J = 7.57 Hz, 3 H), 2.29 (s, 3 H), 3.80 (s, 3 H), 5.11 (q, J = 7.35 Hz, 1 H), 6.99 (d, J = 8.83 Hz, 2 H), 7.20 (s, 1 H), 7.36 (dd, J = 8.83 Hz, 2 H), 7.58 (m, 2 H), 7.61 (s, 1 H), 7.68 (m, 2 H) LC-MS (method 2) Rt = 1.16 min MS (ESIpos): m/z = 414.5 [M + H]+ 49% yield 2-(4-fluoro-N-[5-(4- methoxybenzoyl)-4- methyl-thiazol-2- yl]anilino)propanamide (enantiomer 2) Chiral HPLC Example 176.2 HPLC separation of rac-2-(4-fluoro-N-[5-(4-methoxybenzoyl)-4-methyl-thiazol-2- yl]anilino)propanamide(80 mg, 0.19 mmol, Example 176 on a chiral column gave 31 mg (39% yield) of 2-(4-fluoro-N-[5-(4-methoxybenzoyl)-4-methyl-thiazol-2-yl]anilino)propanamide, enantiomer 2.
Article Title: Substituted aminothiazoles as DGKzeta inhibitors for immune activation
Article Snippet: Preparative chiral HPLC Instrument: Sepiatec: Prep SFC100; Column: Chiralpak IB 5μ 250 × 30 mm; eluent A: CO2; eluent B: 2-propanol + 0.4 vol % diethylamine; isocratic: 35% B; flow: 100 mL/min; temperature: 40° C.; BPR: 150 bar; UV: 254 nm Analytical chiral HPLC: Rt = 4.42 min Instrument: Agilent: 1260, Aurora SFC-Modul; Column: Chiralpak IB 5μ 100 × 4.6 mm; eluent A: CO2; eluent B: 2-propanol + 0.4 vol % diethylamine; isocratic: 35% B; flow: 4 mL/min; temperature: 37.5° C.; BPR: 100 bar; UV: 254 nm 171 Intermediate 111, rac-2- bromopropanamide 1H NMR (400 MHz, DMSO-d6) δ ppm = 1.16 (d, J = 7.35 Hz, 3 H), 5.07 (q, J = 7.35 Hz, 1 H), 7.29 (m, 5 H), 7.58 (m, 1 H), 7.63 (m, 2 H), 7.76 (m, 2 H), 8.21 (m, 2 H) Biotage (method X) LC-MS (method 2) Rt = 1.21 min MS (ESIpos): m/z = 511.0 [M + H]+ 39% yield rac-2-(N-[4-amino-5-(4- iodobenzoyl)thiazol-2-yl] 4-fluoro- anilino)propanamide 172 Intermediate 113, rac-2- bromopropanamide 1H NMR (400 MHz, DMSO-d6) δ ppm = 1.20 (d, J = 7.35 Hz, 3 H), 3.87 (s, 3 H), 5.03 (q, J = 7.35 Hz, 1 H), 6.85 (dd, J = 8.62, 0.76 Hz, 1 H), 7.31 (s, 1 H), 7.52 (m, 1 H), 7.64 (s, 1 H), 7.76 (m, 2 H), 7.84 (dd, J = 8.49, 2.41 Hz, 1 H), 8.14 (m, 2 H), 8.37 (m, 1 H) Biotage (method Y) LC-MS (method 2) Rt = 1.14 min MS (ESIpos): m/z = 450.3 [M + H]+ 38% yield rac-2-(N-[4-amino-5-(6- methoxypyridine-3- carbonyl)thiazol-2-yl]-4- chloro-3-fluoro- anilino)propanamide 172.1 (R)-2-(N-[4-amino-5-(6- and methoxypyridine-3- 172.2 carbonyl)thiazol-2-yl]-4- chloro-3-fluoro- anilino)propanamide and (S)-2-(N-[4-amino-5-(6- methoxypyridine-3- carbonyl)thiazol-2-yl]-4- chloro-3-fluoro- anilino)propanamide 172.1 Example 172 1H NMR (400 MHz, DMSO-d6) δ ppm = 1.20 (d, J = 7.35 Hz, 3 H), 3.87 (s, 3 H), 5.03 (q, J = 7.35 Hz, 1 H), 6.85 (dd, J = 8.62, 0.76 Hz, 1 H), 7.31 (s, 1 H), 7.52 (m, 1 H), 7.64 (s, 1 H), 7.76 (m, 2 H), 7.84 (dd, J = 8.49, 2.41 Hz, 1 H), 8.14 (m, 2 H), 8.37 (m, 1 H) LC-MS (method 2) Rt = 1.14 min MS (ESIpos): m/z = 450.3 [M + H]+ 24% yield 2-(N-[4-amino-5-(6- methoxypyridine-3- carbonyl)thiazol-2-yl]-4- chloro-3-fluoro- anilino)propanamide (enantiomer 1) Chiral HPLC Example 172.1 HPLC separation of rac-2-(N-[4-amino-5-(6-methoxypyridine-3-carbonyl)thiazol-2-yl]-4-chloro-3- fluoro-anilino)propanamide(312 mg, 0.69 mmol, Example 172) on a chiral column gave 78 mg (24% yield) of 2-(N-[4-amino-5-(6-methoxypyridine-3-carbonyl)thiazol-2-yl]-4-chloro-3-fluoro- anilino)propanamide, enantiomer 1. .. Preparative chiral HPLC Instrument: Sepiatec: Prep SFC100; Column: Chiralpak IG 5μ 250 × 30 mm; eluent A: CO2; eluent B: methanol + 0.2 vol % aqueous ammonia (32%); isocratic: 30% B; flow: 100 mL/min; temperature: 40° C.; BPR: 150 bar; UV: 254 nm Analytical chiral HPLC: Rt = 2.61 min Instrument: Agilent: 1260, Aurora SFC-Modul; Column: Chiralpak IG 5μ 100 × 4.6 mm; eluent A: CO2; eluent B: methanol + 0.2 vol % aqueous ammonia (32%); isocratic: 30% B; flow: 4 mL/min; temperature: 37.5° C.; BPR: 100 bar; UV: 254 nm 172.2 Example 172 1H NMR (400 MHz, DMSO-d6) δ ppm = 1.20 (d, J = 7.35 Hz, 3 H), 3.87 (s, 3 H), 5.03 (q, J = 7.35 Hz, 1 H), 6.85 (dd, J = 8.62, 0.76 Hz, 1 H), 7.31 (s, 1 H), 7.52 (m, 1 H), 7.64 (s, 1 H), 7.76 (m, 2 H), 7.84 (dd, J = 8.49, 2.41 Hz, 1 H), 8.14 (m, 2 H), 8.37 (m, 1 H) LC-MS (method 2) Rt = 1.14 min MS (ESIpos): m/z = 450.3 [M + H]+ 23% yield 2-(N-[4-amino-5-(6- methoxypyridine-3- carbonyl)thiazol-2-yl]-4- chloro-3-fluoro- anilino)propanamide (enantiomer 2) Chiral HPLC Example 172.2 HPLC separation of rac-2-(N-[4-amino-5-(6-methoxypyridine-3-carbonyl)thiazol-2-yl]-4-chloro-3- fluoro-anilino)propanamide(312 mg, 0.69 mmol, Example 172) on a chiral column gave 73 mg (23% yield) of 2-(N-[4-amino-5-(6-methoxypyridine-3-carbonyl)thiazol-2-yl]-4-chloro-3-fluoro- anilino)propanamide, enantiomer 2. .. Preparative chiral HPLC Instrument: Sepiatec: Prep SFC100; Column: Chiralpak IG 5μ 250 × 30 mm; eluent A: CO2; eluent B: methanol + 0.2 vol % aqueous ammonia (32%); isocratic: 30% B; flow: 100 mL/min; temperature: 40° C.; BPR: 150 bar UV: 254 nm Analytical chiral HPLC: Rt = 3.40 min Instrument: Agilent: 1260, Aurora SFC-Modul; Column: Chiralpak IG 5μ 100 × 4.6 mm; eluent A: CO2; eluent B: methanol + 0.2 vol % aqueous ammonia (32%); isocratic: 30% B; flow: 4 mL/min; temperature: 37.5° C.; BPR: 100 bar; UV: 254 nm 173 Intermediate 114, rac-2- bromopropanamide 1H NMR (400 MHz, DMSO-d6) δ ppm = 1.16 (d, J = 7.35 Hz, 3 H), 5.07 (m, 1 H), 6.94 (m, 2 H), 7.06 (m, 2 H), 7.20 (m, 1 H), 7.25 (s, 1 H), 7.33 (dd, J = 8.87 Hz, 2 H), 7.41 (m, 2 H), 7.53 (m, 2 H), 7.58 (s, 1 H), 7.64 (m, 2 H), 8.14 (m, 2 H) Biotage (method X) LC-MS (method 2) Rt = 1.26 min MS (ESIpos): m/z = 477.2 [M + H]+ 56% yield rac-2-(N-[4-amino-5-(4- phenoxybenzoyl)thiazol-2- yl]-4-fluoro- anilino)propanamide 174 Intermediate 115, rac-2- bromopropanamide 1H NMR (400 MHz, DMSO-d6) δ ppm = 1.15 (d, J = 7.35 Hz, 3 H), 3.73 (s, 3H), 5.06 (q, J = 7.35 Hz, 1 H), 7.01 (d, J = 9.13 Hz, 2 H), 7.16 (m, 2 H), 7.21 (s, 1 H), 7.27 (t, J = 72 Hz, 1 H), 7.46 (m, 2 H), 7.54 (m, 3 H), 8.15 (m, 2 H) Biotage (method X) LC-MS (method 2) Rt = 1.08 min MS (ESIpos): m/z = 463.2 [M + H]+ 77% yield rac-2-(N-[4-amino-5-[4- (difluoromethoxy)benzoyl] thiazol-2-yl]-4-methoxy- anilino)propanamide 174.1 (R)-2-(N-[4-amino-5-[4- and (difluoromethoxy)benzoyl] 174.2 thiazol-2-yl]-4-methoxy- anilino)propanamide and (S)-2-(N-[4-amino-5-[4- (difluoromethoxy)benzoyl] thiazol-2-yl]-4-methoxy- anilino)propanamide 174.1 Example 174 1H NMR (400 MHz, DMSO-d6) δ ppm = 1.15 (d, J = 7.35 Hz, 3 H), 3.73 (s, 3H), 5.06 (q, J = 7.35 Hz, 1 H), 7.01 (d, J = 9.13 Hz, 2 H), 7.16 (m, 2 H), 7.21 (s, 1 H), 7.27 (t, J = 72 Hz, 1 H), 7.46 (m, 2 H), 7.54 (m, 3 H), 8.15 (m, 2 H) LC-MS (method 2) Rt = 1.08 min MS (ESIpos): m/z = 463.2 [M + H]+ 31% yield 2-(N-[4-amino-5-[4- (difluoromethoxy)benzoyl] thiazol-2-yl]-4-methoxy- anilino)propanamide (enantiomer 1) Chiral HPLC Example 174.1 HPLC separation of rac-2-(N-[4-amino-5-[4-(difluoromethoxy)benzoyl]thiazol-2-yl]-4-methoxy- anilino)propanamide(316 mg, 0.66 mmol, Example 174 on a chiral column gave 121 mg (31% yield) of 2-(N-[4-amino-5-[4-(difluoromethoxy)benzoyl]thiazol-2-yl]-4-methoxy- anilino)propanamide, enantiomer 1.
Article Title: Substituted aminothiazoles as DGKzeta inhibitors for immune activation
Article Snippet: Preparative chiral HPLC Instrument: Sepiatec: Prep SFC100; Column: Chiralpak IG 5μ 250 × 30 mm; eluent A: CO2; eluent B: ethanol + 0.2 vol % aqueous ammonia (32%); isocratic: 15% B; flow: 100 mL/min; temperature: 40° C.; BPR: 150 bar; UV: 254 nm Analytical chiral HPLC: Rt = 2.81 min Instrument: Agilent: 1260, Aurora SFC-Modul; Column: Chiralpak IG 5μ 100 × 4.6 mm; eluent A: CO2; eluent B: ethanol + 0.2 vol % aqueous ammonia (32%); isocratic: 25% B; flow: 4 mL/min; temperature: 37.5° C.; BPR: 100 bar; UV: 254 nm 187.2 Example 187 1H NMR (400 MHz, DMSO-d6) δ ppm = 1.15 (d, J = 7.60 Hz, 3 H), 2.45 (s, 3 H), 5.06 (br s, 1 H), 7.10-7.23 (m, 1 H), 7.23-7.28 (m, 2 H), 7.33 (dd, J = 8.74 Hz, 2 H), 7.59 (br s, 1 H), 7.63 (dd, J = 8.87, 5.07 Hz, 2 H), 8.05-8.42 (m, 2 H), 8.46 (d, J = 5.07 Hz, 1 H) 23% yield 2-(N-[4-amino-5-(2- methylpyridine-4- carbonyl)thiazol-2-yl]-4- fluoro-anilino)propanamide (enantiomer 2) Chiral HPLC Example 187.2 HPLC separation of rac-2-(N-[4-amino-5-(2-methylpyridine-4-carbonyl)thiazol-2-yl]-4-fluoro- anilino)propanamide(333 mg, 0.83 mmol, Example 187) on a chiral column gave 78 mg (23% yield) of 2-(N-[4-amino-5-(2-methylpyridine-4-carbonyl)thiazol-2-yl]-4-fluoro-anilino)propanamide, enantiomer 2. .. Preparative chiral HPLC Instrument: Sepiatec: Prep SFC100; Column: Chiralpak IG 5μ 250 × 30 mm; eluent A: CO2; eluent B: ethanol + 0.2 vol % aqueous ammonia (32%); isocratic: 15% B; flow: 100 mL/min; temperature: 40° C.; BPR: 150 bar; UV: 254 nm Analytical chiral HPLC: Rt = 3.56 min Instrument: Agilent: 1260, Aurora SFC-Modul; Column: Chiralpak IG 5μ 100 × 4.6 mm; eluent A: CO2; eluent B: ethanol + 0.2 vol % aqueous ammonia (32%); isocratic: 25% B; flow: 4 mL/min; temperature: 37.5° C.; BPR: 100 bar; UV: 254 nm 188 Intermediate 129, rac-2- bromopropanamide 1H NMR (400 MHz, DMSO-d6) δ ppm = 1.16 (d, J = 7.35 Hz, 3 H), 4.99-5.13 (m, 1 H), 6.97 (t, J = 52 Hz, 1 H), 7.27 (s, 1 H), 7.33 (t J = 8.87 Hz, 2 H), 7.56-7.61 (m, 2 H), 7.64 (dd, J = 8.87, 5.07 Hz, 2 H), 7.67 (s, 1 H), 8.20-8.57 (m, 2 H), 8.73 (d, J = 4.56 Hz, 1 H) RP-HPLC (method C basic) LC-MS (method 2) Rt = 1.00 min MS (ESIpos): m/z = 436.3 [M + H]+ 13% yield rac-2-(N-[4-amino-5-[2- (difluoromethyl)pyridine-4- carbonyl]thiazol-2-yl]-4- fluoro-anilino)propanamide 189 Intermediate 130, rac-2- bromopropanamide 1H NMR (400 MHz, DMSO-d6) δ ppm = 1.22 (d, J = 7.35 Hz, 3 H), 5.05 (q, J = 7.35 Hz, 1 H), 5.74 (s, 1 H), 7.32 (s, 1 H), 7.37-7.44 (m, 2 H), 7.48-7.55 (m, 2 H), 7.56-7.65 (m, 3 H), 8.06-8.42 (m, 2 H), 8.68-8.73 (m, 2 H) RP-HPLC (method C basic) LC-MS (method 2) Rt = 0.80 min MS (ESIpos): m/z = 368.2 [M + H]+ 17% yield rac-2-[(4-amino-5-benzoyl- thiazol-2-yl)-(4- pyridyl)amino]propanamide 190 Intermediate 131, rac-2- bromopropanamide 1H NMR (400 MHz, DMSO-d6) δ ppm = 1.15 (d, J = 7.35 Hz, 3 H), 3.83 (s, 3H), 5.00-5.11 (m, 1 H), 6.76 (s, 1 H), 6.99 (dd, J = 5.20, 1.39 Hz, 1 H), 7.26 (s, 1 H), 7.30- 7.37 (m, 2 H), 7.59 (s, 1 H), 7.63 (dd, J = 8.87, 5.07 Hz, 2 H), 8.17-8.20 (m, 1 H), 8.21-8.48 (m, 2 H) RP-HPLC (method C basic) LC-MS (method 2) Rt = 0.98 min MS (ESIpos): m/z = 416.2 [M + H]+ 29% yield rac-2-(N-[4-amino-5-(2- methoxypyridine-4- carbonyl)thiazol-2-yl]-4- fluoro-anilino)propanamide 191 Intermediate 133; rac-2- bromopropanamide 1H NMR (400 MHz, DMSO-d6) δ ppm = 8.12 (br s, 2H), 7.57 (s, 1H), 7.46-7.53 (m, 3H), 7.34-7.44 (m, 4H), 7.21-7.28 (m, 2H), 4.98- 5.10 (m, 1H), 3.87 (s, 3H), 1.16 (d, J = 7.4 Hz, 3H). ..
Article Title: Substituted aminothiazoles as DGKzeta inhibitors for immune activation
Article Snippet: Preparative chiral HPLC Instrument: Sepiatec: Prep SFC100; Column: Chiralpak IG 5μ 250 × 30 mm; eluent A: CO2; eluent B: methanol + 0.2 vol % aqueous ammonia (32%); isocratic: 30% B; flow: 100 mL/min; temperature: 40° C.; BPR: 150 bar; UV: 254 nm Analytical chiral HPLC: Rt = 2.61 min Instrument: Agilent: 1260, Aurora SFC-Modul; Column: Chiralpak IG 5μ 100 × 4.6 mm; eluent A: CO2; eluent B: methanol + 0.2 vol % aqueous ammonia (32%); isocratic: 30% B; flow: 4 mL/min; temperature: 37.5° C.; BPR: 100 bar; UV: 254 nm 172.2 Example 172 1H NMR (400 MHz, DMSO-d6) δ ppm = 1.20 (d, J = 7.35 Hz, 3 H), 3.87 (s, 3 H), 5.03 (q, J = 7.35 Hz, 1 H), 6.85 (dd, J = 8.62, 0.76 Hz, 1 H), 7.31 (s, 1 H), 7.52 (m, 1 H), 7.64 (s, 1 H), 7.76 (m, 2 H), 7.84 (dd, J = 8.49, 2.41 Hz, 1 H), 8.14 (m, 2 H), 8.37 (m, 1 H) LC-MS (method 2) Rt = 1.14 min MS (ESIpos): m/z = 450.3 [M + H]+ 23% yield 2-(N-[4-amino-5-(6- methoxypyridine-3- carbonyl)thiazol-2-yl]-4- chloro-3-fluoro- anilino)propanamide (enantiomer 2) Chiral HPLC Example 172.2 HPLC separation of rac-2-(N-[4-amino-5-(6-methoxypyridine-3-carbonyl)thiazol-2-yl]-4-chloro-3- fluoro-anilino)propanamide(312 mg, 0.69 mmol, Example 172) on a chiral column gave 73 mg (23% yield) of 2-(N-[4-amino-5-(6-methoxypyridine-3-carbonyl)thiazol-2-yl]-4-chloro-3-fluoro- anilino)propanamide, enantiomer 2. .. Preparative chiral HPLC Instrument: Sepiatec: Prep SFC100; Column: Chiralpak IG 5μ 250 × 30 mm; eluent A: CO2; eluent B: methanol + 0.2 vol % aqueous ammonia (32%); isocratic: 30% B; flow: 100 mL/min; temperature: 40° C.; BPR: 150 bar UV: 254 nm Analytical chiral HPLC: Rt = 3.40 min Instrument: Agilent: 1260, Aurora SFC-Modul; Column: Chiralpak IG 5μ 100 × 4.6 mm; eluent A: CO2; eluent B: methanol + 0.2 vol % aqueous ammonia (32%); isocratic: 30% B; flow: 4 mL/min; temperature: 37.5° C.; BPR: 100 bar; UV: 254 nm 173 Intermediate 114, rac-2- bromopropanamide 1H NMR (400 MHz, DMSO-d6) δ ppm = 1.16 (d, J = 7.35 Hz, 3 H), 5.07 (m, 1 H), 6.94 (m, 2 H), 7.06 (m, 2 H), 7.20 (m, 1 H), 7.25 (s, 1 H), 7.33 (dd, J = 8.87 Hz, 2 H), 7.41 (m, 2 H), 7.53 (m, 2 H), 7.58 (s, 1 H), 7.64 (m, 2 H), 8.14 (m, 2 H) Biotage (method X) LC-MS (method 2) Rt = 1.26 min MS (ESIpos): m/z = 477.2 [M + H]+ 56% yield rac-2-(N-[4-amino-5-(4- phenoxybenzoyl)thiazol-2- yl]-4-fluoro- anilino)propanamide 174 Intermediate 115, rac-2- bromopropanamide 1H NMR (400 MHz, DMSO-d6) δ ppm = 1.15 (d, J = 7.35 Hz, 3 H), 3.73 (s, 3H), 5.06 (q, J = 7.35 Hz, 1 H), 7.01 (d, J = 9.13 Hz, 2 H), 7.16 (m, 2 H), 7.21 (s, 1 H), 7.27 (t, J = 72 Hz, 1 H), 7.46 (m, 2 H), 7.54 (m, 3 H), 8.15 (m, 2 H) Biotage (method X) LC-MS (method 2) Rt = 1.08 min MS (ESIpos): m/z = 463.2 [M + H]+ 77% yield rac-2-(N-[4-amino-5-[4- (difluoromethoxy)benzoyl] thiazol-2-yl]-4-methoxy- anilino)propanamide 174.1 (R)-2-(N-[4-amino-5-[4- and (difluoromethoxy)benzoyl] 174.2 thiazol-2-yl]-4-methoxy- anilino)propanamide and (S)-2-(N-[4-amino-5-[4- (difluoromethoxy)benzoyl] thiazol-2-yl]-4-methoxy- anilino)propanamide 174.1 Example 174 1H NMR (400 MHz, DMSO-d6) δ ppm = 1.15 (d, J = 7.35 Hz, 3 H), 3.73 (s, 3H), 5.06 (q, J = 7.35 Hz, 1 H), 7.01 (d, J = 9.13 Hz, 2 H), 7.16 (m, 2 H), 7.21 (s, 1 H), 7.27 (t, J = 72 Hz, 1 H), 7.46 (m, 2 H), 7.54 (m, 3 H), 8.15 (m, 2 H) LC-MS (method 2) Rt = 1.08 min MS (ESIpos): m/z = 463.2 [M + H]+ 31% yield 2-(N-[4-amino-5-[4- (difluoromethoxy)benzoyl] thiazol-2-yl]-4-methoxy- anilino)propanamide (enantiomer 1) Chiral HPLC Example 174.1 HPLC separation of rac-2-(N-[4-amino-5-[4-(difluoromethoxy)benzoyl]thiazol-2-yl]-4-methoxy- anilino)propanamide(316 mg, 0.66 mmol, Example 174 on a chiral column gave 121 mg (31% yield) of 2-(N-[4-amino-5-[4-(difluoromethoxy)benzoyl]thiazol-2-yl]-4-methoxy- anilino)propanamide, enantiomer 1. .. Preparative chiral HPLC Instrument: PrepCon Labomatic HPLC-3; Column: YMC Amylose SA 10μ, 250 × 50; eluent A: methyl tert-butyl ether + 0.1 vol % diethylamine; eluent B: methanol; isocratic: 50% A + 50% B; flow: 100 mL/min; temperature: 25° C.; UV: 254 nm Analytical chiral HPLC: Rt = 1.28 min Instrument: Thermo Fisher UltiMate 3000; Column: YMC Amylose SA 3μ, 100 × 4.6; eluent A: methyl tert-butyl ether + 0.1 vol % diethylamine; eluent B: acetonitrile; isocratic: 50% A + 50% B; flow: 1.4 mL/min; temperature: 25° C.; UV: 254 nm 174.2 Example 174 1H NMR (400 MHz, DMSO-d6) δ ppm = 1.15 (d, J = 7.35 Hz, 3 H), 3.73 (s, 3H), 5.06 (q, J = 7.35 Hz, 1 H), 7.01 (d, J = 9.13 Hz, 2 H), 7.16 (m, 2 H), 7.21 (s, 1 H), 7.27 (t, J = 72 Hz, 1 H), 7.46 (m, 2 H), 7.54 (m, 3 H), 8.15 (m, 2 H) LC-MS (method 2) Rt = 1.08 min MS (ESIpos): m/z = 463.2 [M + H]+ 31% yield 2-(N-[4-amino-5-[4- (difluoromethoxy)benzoyl] thiazol-2-yl]-4-methoxy- anilino)propanamide (enantiomer 2) Chiral HPLC Example 174.2 HPLC separation of rac-2-(N-[4-amino-5-[4-(difluoromethoxy)benzoyl]thiazol-2-yl]-4-methoxy- anilino)propanamide(316 mg, 0.66 mmol, Example 174 on a chiral column gave 121 mg (31% yield) of 2-(N-[4-amino-5-[4-(difluoromethoxy)benzoyl]thiazol-2-yl]-4-methoxy- anilino)propanamide, enantiomer 2.
Liquid Chromatography with Mass Spectroscopy:Article Title: Substituted aminothiazoles as DGKzeta inhibitors for immune activation
Article Snippet: Preparative chiral HPLC Instrument: PrepCon Labomatic HPLC-3; Column: YMC Amylose SA 10μ, 250 × 50; eluent A: methyl tert-butyl ether + 0.1 vol % diethylamine; eluent B: methanol; isocratic: 50% A + 50% B; flow: 100 mL/min; temperature: 25° C.; UV: 254 nm Analytical chiral HPLC: Rt = 1.28 min Instrument: Thermo Fisher UltiMate 3000; Column: YMC Amylose SA 3μ, 100 × 4.6; eluent A: methyl tert-butyl ether + 0.1 vol % diethylamine; eluent B: acetonitrile; isocratic: 50% A + 50% B; flow: 1.4 mL/min; temperature: 25° C.; UV: 254 nm 174.2 Example 174 1H NMR (400 MHz, DMSO-d6) δ ppm = 1.15 (d, J = 7.35 Hz, 3 H), 3.73 (s, 3H), 5.06 (q, J = 7.35 Hz, 1 H), 7.01 (d, J = 9.13 Hz, 2 H), 7.16 (m, 2 H), 7.21 (s, 1 H), 7.27 (t, J = 72 Hz, 1 H), 7.46 (m, 2 H), 7.54 (m, 3 H), 8.15 (m, 2 H) LC-MS (method 2) Rt = 1.08 min MS (ESIpos): m/z = 463.2 [M + H]+ 31% yield 2-(N-[4-amino-5-[4- (difluoromethoxy)benzoyl] thiazol-2-yl]-4-methoxy- anilino)propanamide (enantiomer 2) Chiral HPLC Example 174.2 HPLC separation of rac-2-(N-[4-amino-5-[4-(difluoromethoxy)benzoyl]thiazol-2-yl]-4-methoxy- anilino)propanamide(316 mg, 0.66 mmol, Example 174 on a chiral column gave 121 mg (31% yield) of 2-(N-[4-amino-5-[4-(difluoromethoxy)benzoyl]thiazol-2-yl]-4-methoxy- anilino)propanamide, enantiomer 2. .. Preparative chiral HPLC Instrument: PrepCon Labomatic HPLC-3; Column: YMC Amylose SA 10μ, 250 × 50; eluent A: methyl tert-butyl ether + 0.1 vol % diethylamine; eluent B: methanol; isocratic: 50% A + 50% B; flow: 100 mL/min; temperature: 25° C.; UV: 254 nm Analytical chiral HPLC: Rt = 1.62 min Instrument: Thermo Fisher UltiMate 3000; Column: YMC Amylose SA 3μ, 100 × 4.6; eluent A: methyl tert-butyl ether + 0.1 vol % diethylamine; eluent B: acetonitrile; isocratic: 50% A + 50% B; flow: 1.4 mL/min; temperature: 25° C.; UV: 254 nm 175 Intermediate 116, rac-2- bromopropanamide 1H NMR (400 MHz, DMSO-d6) δ ppm = 1.16 (d, J = 7.35 Hz, 3 H), 5.07 (q, J = 7.35 Hz, 1 H), 7.26 (br s, 1 H), 7.33 (m, 3 H), 7.59 (br s, 1 H), 7.63 (m, 3 H), 7.71 (m, 2 H), 8.23 (m, 3 H), 8.38 (m, 1 H) Biotage (method X) LC-MS (method 2) Rt = 1.05 min MS (ESIpos): m/z = 430.1 [M + H]+ 66% yield rac-2-(N-[4-amino-5-(4- nitrobenzoyl)thiazol-2-yl]- 4-fluoro- anilino)propanamide 176 Intermediate 117, rac-2- bromopropanamide 1H NMR (500 MHz, DMSO-d6) δ ppm = 1.16 (d, J = 7.57 Hz, 3 H), 2.29 (s, 3 H), 3.80 (s, 3 H), 5.11 (q, J = 7.35 Hz, 1 H), 6.99 (d, J = 8.83 Hz, 2 H), 7.20 (s, 1 H), 7.36 (dd, J = 8.83 Hz, 2 H), 7.58 (m, 2 H), 7.61 (s, 1 H), 7.68 (m, 2 H) LC-MS (method 2) Rt = 1.16 min MS (ESIpos): m/z = 414.5 [M + H]+ 79% yield rac-2-(4-fluoro-N-[5-(4- methoxybenzoyl)-4- methyl-thiazol-2- yl]anilino)propanamide 176.1 (R)-2-(4-fluoro-N-[5-(4- and methoxybenzoyl)-4- 176.2 methyl-thiazol-2- yl]anilino)propanamide and (S)-2-(4-fluoro-N-[5- (4-methoxybenzoy)-4- methyl-thiazol-2- yl]anilino)propanamide 176.1 Example 176 1H NMR (500 MHz, DMSO-d6) δ ppm = 1.16 (d, J = 7.57 Hz, 3 H), 2.29 (s, 3 H), 3.80 (s, 3 H), 5.11 (q, J = 7.35 Hz, 1 H), 6.99 (d, J = 8.83 Hz, 2 H), 7.20 (s, 1 H), 7.36 (dd, J = 8.83 Hz, 2 H), 7.58 (m, 2 H), 7.61 (s, 1 H), 7.68 (m, 2 H) LC-MS (method 2) Rt = 1.16 min MS (ESIpos): m/z = 414.5 [M + H]+ 49% yield 2-(4-fluoro-N-[5-(4- methoxybenzoyl)-4- methyl-thiazol-2- yl]anilino)propanamide (enantiomer 1) Chiral HPLC Example 176.1 HPLC separation of rac-2-(4-fluoro-N-[5-(4-methoxybenzoyl)-4-methyl-thiazol-2- yl]anilino)propanamide(80 mg, 0.19 mmol, Example 176 on a chiral column gave 39 mg (49% yield) of 2-(4-fluoro-N-[5-(4-methoxybenzoyl)-4-methyl-thiazol-2-yl]anilino)propanamide, enantiomer 1. .. Preparative chiral HPLC Instrument: PrepCon Labomatic HPLC-4; Column: YMC Cellulose SC 10μ, 250 × 50; eluent A: hexane + 0.1 vol % diethylamine; eluent B: ethanol; isocratic: 80% A + 20% B; flow: 100 mL/min; temperature: 25° C.; UV: 254 nm Analytical chiral HPLC: Rt = 4.20 min Instrument: Thermo Fisher UltiMate 3000; Column: YMC Cellulose SC 3μ, 100 × 4.6; eluent A: hexane + 0.1 vol % diethylamine; eluent B: ethanol; isocratic: 80% A + 20% B; flow: 1.4 mL/min; temperature: 25° C.; UV: 254 nm 176.2 Example 176 1H NMR (500 MHz, DMSO-d6) δ ppm = 1.16 (d, J = 7.57 Hz, 3 H), 2.29 (s, 3 H), 3.80 (s, 3 H), 5.11 (q, J = 7.35 Hz, 1 H), 6.99 (d, J = 8.83 Hz, 2 H), 7.20 (s, 1 H), 7.36 (dd, J = 8.83 Hz, 2 H), 7.58 (m, 2 H), 7.61 (s, 1 H), 7.68 (m, 2 H) LC-MS (method 2) Rt = 1.16 min MS (ESIpos): m/z = 414.5 [M + H]+ 49% yield 2-(4-fluoro-N-[5-(4- methoxybenzoyl)-4- methyl-thiazol-2- yl]anilino)propanamide (enantiomer 2) Chiral HPLC Example 176.2 HPLC separation of rac-2-(4-fluoro-N-[5-(4-methoxybenzoyl)-4-methyl-thiazol-2- yl]anilino)propanamide(80 mg, 0.19 mmol, Example 176 on a chiral column gave 31 mg (39% yield) of 2-(4-fluoro-N-[5-(4-methoxybenzoyl)-4-methyl-thiazol-2-yl]anilino)propanamide, enantiomer 2.
Article Title: Substituted aminothiazoles as DGKzeta inhibitors for immune activation
Article Snippet: Preparative chiral HPLC Instrument: Sepiatec: Prep SFC100; Column: Chiralpak IB 5μ 250 × 30 mm; eluent A: CO2; eluent B: 2-propanol + 0.4 vol % diethylamine; isocratic: 35% B; flow: 100 mL/min; temperature: 40° C.; BPR: 150 bar; UV: 254 nm Analytical chiral HPLC: Rt = 4.42 min Instrument: Agilent: 1260, Aurora SFC-Modul; Column: Chiralpak IB 5μ 100 × 4.6 mm; eluent A: CO2; eluent B: 2-propanol + 0.4 vol % diethylamine; isocratic: 35% B; flow: 4 mL/min; temperature: 37.5° C.; BPR: 100 bar; UV: 254 nm 171 Intermediate 111, rac-2- bromopropanamide 1H NMR (400 MHz, DMSO-d6) δ ppm = 1.16 (d, J = 7.35 Hz, 3 H), 5.07 (q, J = 7.35 Hz, 1 H), 7.29 (m, 5 H), 7.58 (m, 1 H), 7.63 (m, 2 H), 7.76 (m, 2 H), 8.21 (m, 2 H) Biotage (method X) LC-MS (method 2) Rt = 1.21 min MS (ESIpos): m/z = 511.0 [M + H]+ 39% yield rac-2-(N-[4-amino-5-(4- iodobenzoyl)thiazol-2-yl] 4-fluoro- anilino)propanamide 172 Intermediate 113, rac-2- bromopropanamide 1H NMR (400 MHz, DMSO-d6) δ ppm = 1.20 (d, J = 7.35 Hz, 3 H), 3.87 (s, 3 H), 5.03 (q, J = 7.35 Hz, 1 H), 6.85 (dd, J = 8.62, 0.76 Hz, 1 H), 7.31 (s, 1 H), 7.52 (m, 1 H), 7.64 (s, 1 H), 7.76 (m, 2 H), 7.84 (dd, J = 8.49, 2.41 Hz, 1 H), 8.14 (m, 2 H), 8.37 (m, 1 H) Biotage (method Y) LC-MS (method 2) Rt = 1.14 min MS (ESIpos): m/z = 450.3 [M + H]+ 38% yield rac-2-(N-[4-amino-5-(6- methoxypyridine-3- carbonyl)thiazol-2-yl]-4- chloro-3-fluoro- anilino)propanamide 172.1 (R)-2-(N-[4-amino-5-(6- and methoxypyridine-3- 172.2 carbonyl)thiazol-2-yl]-4- chloro-3-fluoro- anilino)propanamide and (S)-2-(N-[4-amino-5-(6- methoxypyridine-3- carbonyl)thiazol-2-yl]-4- chloro-3-fluoro- anilino)propanamide 172.1 Example 172 1H NMR (400 MHz, DMSO-d6) δ ppm = 1.20 (d, J = 7.35 Hz, 3 H), 3.87 (s, 3 H), 5.03 (q, J = 7.35 Hz, 1 H), 6.85 (dd, J = 8.62, 0.76 Hz, 1 H), 7.31 (s, 1 H), 7.52 (m, 1 H), 7.64 (s, 1 H), 7.76 (m, 2 H), 7.84 (dd, J = 8.49, 2.41 Hz, 1 H), 8.14 (m, 2 H), 8.37 (m, 1 H) LC-MS (method 2) Rt = 1.14 min MS (ESIpos): m/z = 450.3 [M + H]+ 24% yield 2-(N-[4-amino-5-(6- methoxypyridine-3- carbonyl)thiazol-2-yl]-4- chloro-3-fluoro- anilino)propanamide (enantiomer 1) Chiral HPLC Example 172.1 HPLC separation of rac-2-(N-[4-amino-5-(6-methoxypyridine-3-carbonyl)thiazol-2-yl]-4-chloro-3- fluoro-anilino)propanamide(312 mg, 0.69 mmol, Example 172) on a chiral column gave 78 mg (24% yield) of 2-(N-[4-amino-5-(6-methoxypyridine-3-carbonyl)thiazol-2-yl]-4-chloro-3-fluoro- anilino)propanamide, enantiomer 1. .. Preparative chiral HPLC Instrument: Sepiatec: Prep SFC100; Column: Chiralpak IG 5μ 250 × 30 mm; eluent A: CO2; eluent B: methanol + 0.2 vol % aqueous ammonia (32%); isocratic: 30% B; flow: 100 mL/min; temperature: 40° C.; BPR: 150 bar; UV: 254 nm Analytical chiral HPLC: Rt = 2.61 min Instrument: Agilent: 1260, Aurora SFC-Modul; Column: Chiralpak IG 5μ 100 × 4.6 mm; eluent A: CO2; eluent B: methanol + 0.2 vol % aqueous ammonia (32%); isocratic: 30% B; flow: 4 mL/min; temperature: 37.5° C.; BPR: 100 bar; UV: 254 nm 172.2 Example 172 1H NMR (400 MHz, DMSO-d6) δ ppm = 1.20 (d, J = 7.35 Hz, 3 H), 3.87 (s, 3 H), 5.03 (q, J = 7.35 Hz, 1 H), 6.85 (dd, J = 8.62, 0.76 Hz, 1 H), 7.31 (s, 1 H), 7.52 (m, 1 H), 7.64 (s, 1 H), 7.76 (m, 2 H), 7.84 (dd, J = 8.49, 2.41 Hz, 1 H), 8.14 (m, 2 H), 8.37 (m, 1 H) LC-MS (method 2) Rt = 1.14 min MS (ESIpos): m/z = 450.3 [M + H]+ 23% yield 2-(N-[4-amino-5-(6- methoxypyridine-3- carbonyl)thiazol-2-yl]-4- chloro-3-fluoro- anilino)propanamide (enantiomer 2) Chiral HPLC Example 172.2 HPLC separation of rac-2-(N-[4-amino-5-(6-methoxypyridine-3-carbonyl)thiazol-2-yl]-4-chloro-3- fluoro-anilino)propanamide(312 mg, 0.69 mmol, Example 172) on a chiral column gave 73 mg (23% yield) of 2-(N-[4-amino-5-(6-methoxypyridine-3-carbonyl)thiazol-2-yl]-4-chloro-3-fluoro- anilino)propanamide, enantiomer 2. .. Preparative chiral HPLC Instrument: Sepiatec: Prep SFC100; Column: Chiralpak IG 5μ 250 × 30 mm; eluent A: CO2; eluent B: methanol + 0.2 vol % aqueous ammonia (32%); isocratic: 30% B; flow: 100 mL/min; temperature: 40° C.; BPR: 150 bar UV: 254 nm Analytical chiral HPLC: Rt = 3.40 min Instrument: Agilent: 1260, Aurora SFC-Modul; Column: Chiralpak IG 5μ 100 × 4.6 mm; eluent A: CO2; eluent B: methanol + 0.2 vol % aqueous ammonia (32%); isocratic: 30% B; flow: 4 mL/min; temperature: 37.5° C.; BPR: 100 bar; UV: 254 nm 173 Intermediate 114, rac-2- bromopropanamide 1H NMR (400 MHz, DMSO-d6) δ ppm = 1.16 (d, J = 7.35 Hz, 3 H), 5.07 (m, 1 H), 6.94 (m, 2 H), 7.06 (m, 2 H), 7.20 (m, 1 H), 7.25 (s, 1 H), 7.33 (dd, J = 8.87 Hz, 2 H), 7.41 (m, 2 H), 7.53 (m, 2 H), 7.58 (s, 1 H), 7.64 (m, 2 H), 8.14 (m, 2 H) Biotage (method X) LC-MS (method 2) Rt = 1.26 min MS (ESIpos): m/z = 477.2 [M + H]+ 56% yield rac-2-(N-[4-amino-5-(4- phenoxybenzoyl)thiazol-2- yl]-4-fluoro- anilino)propanamide 174 Intermediate 115, rac-2- bromopropanamide 1H NMR (400 MHz, DMSO-d6) δ ppm = 1.15 (d, J = 7.35 Hz, 3 H), 3.73 (s, 3H), 5.06 (q, J = 7.35 Hz, 1 H), 7.01 (d, J = 9.13 Hz, 2 H), 7.16 (m, 2 H), 7.21 (s, 1 H), 7.27 (t, J = 72 Hz, 1 H), 7.46 (m, 2 H), 7.54 (m, 3 H), 8.15 (m, 2 H) Biotage (method X) LC-MS (method 2) Rt = 1.08 min MS (ESIpos): m/z = 463.2 [M + H]+ 77% yield rac-2-(N-[4-amino-5-[4- (difluoromethoxy)benzoyl] thiazol-2-yl]-4-methoxy- anilino)propanamide 174.1 (R)-2-(N-[4-amino-5-[4- and (difluoromethoxy)benzoyl] 174.2 thiazol-2-yl]-4-methoxy- anilino)propanamide and (S)-2-(N-[4-amino-5-[4- (difluoromethoxy)benzoyl] thiazol-2-yl]-4-methoxy- anilino)propanamide 174.1 Example 174 1H NMR (400 MHz, DMSO-d6) δ ppm = 1.15 (d, J = 7.35 Hz, 3 H), 3.73 (s, 3H), 5.06 (q, J = 7.35 Hz, 1 H), 7.01 (d, J = 9.13 Hz, 2 H), 7.16 (m, 2 H), 7.21 (s, 1 H), 7.27 (t, J = 72 Hz, 1 H), 7.46 (m, 2 H), 7.54 (m, 3 H), 8.15 (m, 2 H) LC-MS (method 2) Rt = 1.08 min MS (ESIpos): m/z = 463.2 [M + H]+ 31% yield 2-(N-[4-amino-5-[4- (difluoromethoxy)benzoyl] thiazol-2-yl]-4-methoxy- anilino)propanamide (enantiomer 1) Chiral HPLC Example 174.1 HPLC separation of rac-2-(N-[4-amino-5-[4-(difluoromethoxy)benzoyl]thiazol-2-yl]-4-methoxy- anilino)propanamide(316 mg, 0.66 mmol, Example 174 on a chiral column gave 121 mg (31% yield) of 2-(N-[4-amino-5-[4-(difluoromethoxy)benzoyl]thiazol-2-yl]-4-methoxy- anilino)propanamide, enantiomer 1.
Article Title: Substituted aminothiazoles as DGKzeta inhibitors for immune activation
Article Snippet: Preparative chiral HPLC Instrument: Sepiatec: Prep SFC100; Column: Chiralpak IG 5μ 250 × 30 mm; eluent A: CO2; eluent B: ethanol + 0.2 vol % aqueous ammonia (32%); isocratic: 15% B; flow: 100 mL/min; temperature: 40° C.; BPR: 150 bar; UV: 254 nm Analytical chiral HPLC: Rt = 2.81 min Instrument: Agilent: 1260, Aurora SFC-Modul; Column: Chiralpak IG 5μ 100 × 4.6 mm; eluent A: CO2; eluent B: ethanol + 0.2 vol % aqueous ammonia (32%); isocratic: 25% B; flow: 4 mL/min; temperature: 37.5° C.; BPR: 100 bar; UV: 254 nm 187.2 Example 187 1H NMR (400 MHz, DMSO-d6) δ ppm = 1.15 (d, J = 7.60 Hz, 3 H), 2.45 (s, 3 H), 5.06 (br s, 1 H), 7.10-7.23 (m, 1 H), 7.23-7.28 (m, 2 H), 7.33 (dd, J = 8.74 Hz, 2 H), 7.59 (br s, 1 H), 7.63 (dd, J = 8.87, 5.07 Hz, 2 H), 8.05-8.42 (m, 2 H), 8.46 (d, J = 5.07 Hz, 1 H) 23% yield 2-(N-[4-amino-5-(2- methylpyridine-4- carbonyl)thiazol-2-yl]-4- fluoro-anilino)propanamide (enantiomer 2) Chiral HPLC Example 187.2 HPLC separation of rac-2-(N-[4-amino-5-(2-methylpyridine-4-carbonyl)thiazol-2-yl]-4-fluoro- anilino)propanamide(333 mg, 0.83 mmol, Example 187) on a chiral column gave 78 mg (23% yield) of 2-(N-[4-amino-5-(2-methylpyridine-4-carbonyl)thiazol-2-yl]-4-fluoro-anilino)propanamide, enantiomer 2. .. Preparative chiral HPLC Instrument: Sepiatec: Prep SFC100; Column: Chiralpak IG 5μ 250 × 30 mm; eluent A: CO2; eluent B: ethanol + 0.2 vol % aqueous ammonia (32%); isocratic: 15% B; flow: 100 mL/min; temperature: 40° C.; BPR: 150 bar; UV: 254 nm Analytical chiral HPLC: Rt = 3.56 min Instrument: Agilent: 1260, Aurora SFC-Modul; Column: Chiralpak IG 5μ 100 × 4.6 mm; eluent A: CO2; eluent B: ethanol + 0.2 vol % aqueous ammonia (32%); isocratic: 25% B; flow: 4 mL/min; temperature: 37.5° C.; BPR: 100 bar; UV: 254 nm 188 Intermediate 129, rac-2- bromopropanamide 1H NMR (400 MHz, DMSO-d6) δ ppm = 1.16 (d, J = 7.35 Hz, 3 H), 4.99-5.13 (m, 1 H), 6.97 (t, J = 52 Hz, 1 H), 7.27 (s, 1 H), 7.33 (t J = 8.87 Hz, 2 H), 7.56-7.61 (m, 2 H), 7.64 (dd, J = 8.87, 5.07 Hz, 2 H), 7.67 (s, 1 H), 8.20-8.57 (m, 2 H), 8.73 (d, J = 4.56 Hz, 1 H) RP-HPLC (method C basic) LC-MS (method 2) Rt = 1.00 min MS (ESIpos): m/z = 436.3 [M + H]+ 13% yield rac-2-(N-[4-amino-5-[2- (difluoromethyl)pyridine-4- carbonyl]thiazol-2-yl]-4- fluoro-anilino)propanamide 189 Intermediate 130, rac-2- bromopropanamide 1H NMR (400 MHz, DMSO-d6) δ ppm = 1.22 (d, J = 7.35 Hz, 3 H), 5.05 (q, J = 7.35 Hz, 1 H), 5.74 (s, 1 H), 7.32 (s, 1 H), 7.37-7.44 (m, 2 H), 7.48-7.55 (m, 2 H), 7.56-7.65 (m, 3 H), 8.06-8.42 (m, 2 H), 8.68-8.73 (m, 2 H) RP-HPLC (method C basic) LC-MS (method 2) Rt = 0.80 min MS (ESIpos): m/z = 368.2 [M + H]+ 17% yield rac-2-[(4-amino-5-benzoyl- thiazol-2-yl)-(4- pyridyl)amino]propanamide 190 Intermediate 131, rac-2- bromopropanamide 1H NMR (400 MHz, DMSO-d6) δ ppm = 1.15 (d, J = 7.35 Hz, 3 H), 3.83 (s, 3H), 5.00-5.11 (m, 1 H), 6.76 (s, 1 H), 6.99 (dd, J = 5.20, 1.39 Hz, 1 H), 7.26 (s, 1 H), 7.30- 7.37 (m, 2 H), 7.59 (s, 1 H), 7.63 (dd, J = 8.87, 5.07 Hz, 2 H), 8.17-8.20 (m, 1 H), 8.21-8.48 (m, 2 H) RP-HPLC (method C basic) LC-MS (method 2) Rt = 0.98 min MS (ESIpos): m/z = 416.2 [M + H]+ 29% yield rac-2-(N-[4-amino-5-(2- methoxypyridine-4- carbonyl)thiazol-2-yl]-4- fluoro-anilino)propanamide 191 Intermediate 133; rac-2- bromopropanamide 1H NMR (400 MHz, DMSO-d6) δ ppm = 8.12 (br s, 2H), 7.57 (s, 1H), 7.46-7.53 (m, 3H), 7.34-7.44 (m, 4H), 7.21-7.28 (m, 2H), 4.98- 5.10 (m, 1H), 3.87 (s, 3H), 1.16 (d, J = 7.4 Hz, 3H). ..
Article Title: Substituted aminothiazoles as DGKzeta inhibitors for immune activation
Article Snippet: Preparative chiral HPLC Instrument: Sepiatec: Prep SFC100; Column: Chiralpak IG 5μ 250 × 30 mm; eluent A: CO2; eluent B: methanol + 0.2 vol % aqueous ammonia (32%); isocratic: 30% B; flow: 100 mL/min; temperature: 40° C.; BPR: 150 bar; UV: 254 nm Analytical chiral HPLC: Rt = 2.61 min Instrument: Agilent: 1260, Aurora SFC-Modul; Column: Chiralpak IG 5μ 100 × 4.6 mm; eluent A: CO2; eluent B: methanol + 0.2 vol % aqueous ammonia (32%); isocratic: 30% B; flow: 4 mL/min; temperature: 37.5° C.; BPR: 100 bar; UV: 254 nm 172.2 Example 172 1H NMR (400 MHz, DMSO-d6) δ ppm = 1.20 (d, J = 7.35 Hz, 3 H), 3.87 (s, 3 H), 5.03 (q, J = 7.35 Hz, 1 H), 6.85 (dd, J = 8.62, 0.76 Hz, 1 H), 7.31 (s, 1 H), 7.52 (m, 1 H), 7.64 (s, 1 H), 7.76 (m, 2 H), 7.84 (dd, J = 8.49, 2.41 Hz, 1 H), 8.14 (m, 2 H), 8.37 (m, 1 H) LC-MS (method 2) Rt = 1.14 min MS (ESIpos): m/z = 450.3 [M + H]+ 23% yield 2-(N-[4-amino-5-(6- methoxypyridine-3- carbonyl)thiazol-2-yl]-4- chloro-3-fluoro- anilino)propanamide (enantiomer 2) Chiral HPLC Example 172.2 HPLC separation of rac-2-(N-[4-amino-5-(6-methoxypyridine-3-carbonyl)thiazol-2-yl]-4-chloro-3- fluoro-anilino)propanamide(312 mg, 0.69 mmol, Example 172) on a chiral column gave 73 mg (23% yield) of 2-(N-[4-amino-5-(6-methoxypyridine-3-carbonyl)thiazol-2-yl]-4-chloro-3-fluoro- anilino)propanamide, enantiomer 2. .. Preparative chiral HPLC Instrument: Sepiatec: Prep SFC100; Column: Chiralpak IG 5μ 250 × 30 mm; eluent A: CO2; eluent B: methanol + 0.2 vol % aqueous ammonia (32%); isocratic: 30% B; flow: 100 mL/min; temperature: 40° C.; BPR: 150 bar UV: 254 nm Analytical chiral HPLC: Rt = 3.40 min Instrument: Agilent: 1260, Aurora SFC-Modul; Column: Chiralpak IG 5μ 100 × 4.6 mm; eluent A: CO2; eluent B: methanol + 0.2 vol % aqueous ammonia (32%); isocratic: 30% B; flow: 4 mL/min; temperature: 37.5° C.; BPR: 100 bar; UV: 254 nm 173 Intermediate 114, rac-2- bromopropanamide 1H NMR (400 MHz, DMSO-d6) δ ppm = 1.16 (d, J = 7.35 Hz, 3 H), 5.07 (m, 1 H), 6.94 (m, 2 H), 7.06 (m, 2 H), 7.20 (m, 1 H), 7.25 (s, 1 H), 7.33 (dd, J = 8.87 Hz, 2 H), 7.41 (m, 2 H), 7.53 (m, 2 H), 7.58 (s, 1 H), 7.64 (m, 2 H), 8.14 (m, 2 H) Biotage (method X) LC-MS (method 2) Rt = 1.26 min MS (ESIpos): m/z = 477.2 [M + H]+ 56% yield rac-2-(N-[4-amino-5-(4- phenoxybenzoyl)thiazol-2- yl]-4-fluoro- anilino)propanamide 174 Intermediate 115, rac-2- bromopropanamide 1H NMR (400 MHz, DMSO-d6) δ ppm = 1.15 (d, J = 7.35 Hz, 3 H), 3.73 (s, 3H), 5.06 (q, J = 7.35 Hz, 1 H), 7.01 (d, J = 9.13 Hz, 2 H), 7.16 (m, 2 H), 7.21 (s, 1 H), 7.27 (t, J = 72 Hz, 1 H), 7.46 (m, 2 H), 7.54 (m, 3 H), 8.15 (m, 2 H) Biotage (method X) LC-MS (method 2) Rt = 1.08 min MS (ESIpos): m/z = 463.2 [M + H]+ 77% yield rac-2-(N-[4-amino-5-[4- (difluoromethoxy)benzoyl] thiazol-2-yl]-4-methoxy- anilino)propanamide 174.1 (R)-2-(N-[4-amino-5-[4- and (difluoromethoxy)benzoyl] 174.2 thiazol-2-yl]-4-methoxy- anilino)propanamide and (S)-2-(N-[4-amino-5-[4- (difluoromethoxy)benzoyl] thiazol-2-yl]-4-methoxy- anilino)propanamide 174.1 Example 174 1H NMR (400 MHz, DMSO-d6) δ ppm = 1.15 (d, J = 7.35 Hz, 3 H), 3.73 (s, 3H), 5.06 (q, J = 7.35 Hz, 1 H), 7.01 (d, J = 9.13 Hz, 2 H), 7.16 (m, 2 H), 7.21 (s, 1 H), 7.27 (t, J = 72 Hz, 1 H), 7.46 (m, 2 H), 7.54 (m, 3 H), 8.15 (m, 2 H) LC-MS (method 2) Rt = 1.08 min MS (ESIpos): m/z = 463.2 [M + H]+ 31% yield 2-(N-[4-amino-5-[4- (difluoromethoxy)benzoyl] thiazol-2-yl]-4-methoxy- anilino)propanamide (enantiomer 1) Chiral HPLC Example 174.1 HPLC separation of rac-2-(N-[4-amino-5-[4-(difluoromethoxy)benzoyl]thiazol-2-yl]-4-methoxy- anilino)propanamide(316 mg, 0.66 mmol, Example 174 on a chiral column gave 121 mg (31% yield) of 2-(N-[4-amino-5-[4-(difluoromethoxy)benzoyl]thiazol-2-yl]-4-methoxy- anilino)propanamide, enantiomer 1. .. Preparative chiral HPLC Instrument: PrepCon Labomatic HPLC-3; Column: YMC Amylose SA 10μ, 250 × 50; eluent A: methyl tert-butyl ether + 0.1 vol % diethylamine; eluent B: methanol; isocratic: 50% A + 50% B; flow: 100 mL/min; temperature: 25° C.; UV: 254 nm Analytical chiral HPLC: Rt = 1.28 min Instrument: Thermo Fisher UltiMate 3000; Column: YMC Amylose SA 3μ, 100 × 4.6; eluent A: methyl tert-butyl ether + 0.1 vol % diethylamine; eluent B: acetonitrile; isocratic: 50% A + 50% B; flow: 1.4 mL/min; temperature: 25° C.; UV: 254 nm 174.2 Example 174 1H NMR (400 MHz, DMSO-d6) δ ppm = 1.15 (d, J = 7.35 Hz, 3 H), 3.73 (s, 3H), 5.06 (q, J = 7.35 Hz, 1 H), 7.01 (d, J = 9.13 Hz, 2 H), 7.16 (m, 2 H), 7.21 (s, 1 H), 7.27 (t, J = 72 Hz, 1 H), 7.46 (m, 2 H), 7.54 (m, 3 H), 8.15 (m, 2 H) LC-MS (method 2) Rt = 1.08 min MS (ESIpos): m/z = 463.2 [M + H]+ 31% yield 2-(N-[4-amino-5-[4- (difluoromethoxy)benzoyl] thiazol-2-yl]-4-methoxy- anilino)propanamide (enantiomer 2) Chiral HPLC Example 174.2 HPLC separation of rac-2-(N-[4-amino-5-[4-(difluoromethoxy)benzoyl]thiazol-2-yl]-4-methoxy- anilino)propanamide(316 mg, 0.66 mmol, Example 174 on a chiral column gave 121 mg (31% yield) of 2-(N-[4-amino-5-[4-(difluoromethoxy)benzoyl]thiazol-2-yl]-4-methoxy- anilino)propanamide, enantiomer 2.
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