Structure of 2086301-12-2
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| CAS No. : | 2086301-12-2 |
| Formula : | C17H21N3O2S |
| M.W : | 331.43 |
| SMILES Code : | O=C([C@H]1NC[C@H](O)C1)N[C@H](C2=CC=C(C3=C(C)N=CS3)C=C2)C |
| English Name : | (2S,4R)-4-Hydroxy-N-((S)-1-(4-(4-methylthiazol-5-yl)phenyl)ethyl)pyrrolidine-2-carboxamide |
| MDL No. : | MFCD35453547 |
* All experimental methods are cited from the reference, please refer to the original source for details. We do not guarantee the accuracy of the content in the reference.

| Yield | Reaction Conditions | Operation in experiment |
|---|---|---|
| With N-ethyl-N,N-diisopropylamine; N-[(dimethylamino)-3-oxo-1H-1,2,3-triazolo[4,5-b]pyridin-1-yl-methylene]-N-methylmethanaminium hexafluorophosphate In N,N-dimethyl-formamide at 0 - 20℃; for 12h; Inert atmosphere; | ||
| 1.31 g | With N-ethyl-N,N-diisopropylamine; HATU In N,N-dimethyl-formamide at 0 - 20℃; for 12h; Inert atmosphere; | 1 tert- Butyl ((S)-l-((2S,4R)-4-hydroxy-2-(((S)-l-(4-(4-methylthiazol-5- yl)phenyl)ethyl)carb-amoyl)pyrrolidin- 1 -yl)-3 ,3 -dimethyl- 1 -oxobutan-2-yl)carbamate (57) This solid (56), obtained as described above was dissolved in 4N HC1 in dioxane (0327) (25 mL, 100 mmol) and MeOH (25 mL) and the mixture was stirred at ambient temperature for 12 h. The mixture was then concentrated and the residue was dried under vacuum to afford an intermediate, which was used in next step without further purification. UPLC-MS (ESL) calc, for C17H22N3O2S [M+l]+: 332.14, found 332.11. HATU (1.37 g, 3.6 mmol, 1.2 eq) was added to a solution of this intermediate (994 mg, 3.0 mmol, 1.0 eq), (S)-2-((/er/-butoxycarbonyl)amino)-3,3-dimethylbutanoic acid (694 mg, 3.0 mmol, 1.0 eq), and DIPEA (1.57 mL, 9.0 mmol, 3.0 eq) in DMF (10 mL) at 0 °C under N2. The mixture was stirred at ambient temperature for 12 h when TLC showed that the reaction was complete. The reaction mixture was quenched with H2O (100 mL) and extracted with EtOAc (75mLx2). The combined organic layer was washed with brine (100 mL) and dried over Na2S04. The organic solution was filtered and concentrated. The residue was purified by silica gel flash column chromatography with hexane :EtO Ac then DCM: MeOH to afford the desired compound (57) as a white solid (1.31 g, 80% yield). NMR (CDCI3, 400 MHz) d (ppm) 8.65 (s, 1H), 7.70 (d, J = 8.0 Hz, 1H), 7.35- 7.31 (m, 4H), 5.29 (d, J= 9.2 Hz, 1H), 5.06-5.01 (m, 1H), 4.67 (t, J= 8.0 Hz, 1H), 4.46- 4.44 (m, 1H), 4.22-4.19 (m, 1H), 3.91 (d, J = 17.2 Hz, 1H), 3.61-3.58 (m, 1H), 2.46 (s, 3H), 2.37-2.30 (m, 1H), 2.04-1.99 (m, 1H), 1.44 (d, J= 7.2 Hz, 3H), 1.35 (s, 9H), 0.96 (s, 9H); 13C NMR (CDCI3, 100 MHz) d (ppm) 172.22, 170.13, 156.15, 150.56, 148.21, 143.43, 131.74, 130.59, 129.49, 126.46, 80.18, 69.91, 58.86, 56.58, 48.74, 38.60, 36.02, 35.48, 28.34, 26.39, 22.17, 15.95; UPLC-MS (ESF) calc for C28H41N4O5S [M+l]+: 545.28, found 545.35. |
| 510 mg | With N-ethyl-N,N-diisopropylamine; N-[(dimethylamino)-3-oxo-1H-1,2,3-triazolo[4,5-b]pyridin-1-yl-methylene]-N-methylmethanaminium hexafluorophosphate In tetrahydrofuran at 20℃; | F Step F: Add (S)-2-((tert-Butoxycarbonyl)amino)-3,3-dimethylbutyric acid (293 mg, 1.3 mmol), compounds INT-5-5 (420 mg, 1.3 mmol), ), 2-(7-aza-1H-benzotriazol-1-yl)-N,N,N',N'-tetramethyluronium hexafluorophosphate (483 mg, 1.3 mmol) and N-Diisopropylethylamine (818 mg, 6.3 mmol) to tetrahydrofuran (5 ml) solution, respectively, and stir at 20 degrees Celsius for 18 hours. LCMS monitoring shows that raw materials are gone. After quenching the reaction solution with purified water (200 ml), it was extracted with ethyl acetate (100 ml×3). The organic phase was combined and dried with anhydrous sodium sulfate and filtered and concentrated. The resulting mixture was purified by silica gel column chromatography to obtain 510 mg of compound INT-5-6. |
| With N-ethyl-N,N-diisopropylamine; HATU In N,N-dimethyl-formamide at 0 - 20℃; | ||
| With N-ethyl-N,N-diisopropylamine; HATU In N,N-dimethyl-formamide at 0 - 20℃; | 4.1.20. tert-butyl ((S)-1-((2S,4R)-4-hydroxy-2-(((S)-1-(4-(4-methylthiazol-5-yl)phenyl)ethyl) carbamoyl) pyrrolidin-1-yl)-3,3-dimethyl-1-oxobutan-2-yl) carbamate (P13) This solid material (P12), obtained as described above was dissolvedin 2 M HCl in EA (20 mL, 40 mmol), and the mixture was stirred atambient temperature for 12 h. The mixture was then concentrated, andthe residue was dried under vacuum to afford intermediate, which wasused in next step without further purification. HATU (8.50 g, 22.5 mmol,1.4 equiv) was added to a solution of this intermediate (5.80 g, 16.0mmol, 1.0 equiv), (S)-2-((tert-butoxycarbonyl) amino)-3,3- dimethylbutanoicacid (3.46 g, 16.0 mmol, 1.0 equiv), and DIPEA (8.8 mL,48.0 mmol, 3.0 equiv) in DMF (30 mL) at 0 C. The mixture was stirredat ambient temperature for 12 h when TLC showed that the reaction wascomplete. The reaction mixture was quenched with H2O (120 mL) andextracted with EtOAc (75 mL × 2). The combined organic layer waswashed with brine (100 mL) and dried over Na2SO4. The organic solutionwas filtered and concentrated. The residue was purified by silica gelflash column chromatography (DCM: MeOH 30:1) to afford the desiredcompound. | |
| With N-ethyl-N,N-diisopropylamine; HATU In N,N-dimethyl-formamide at 0 - 20℃; | 4.1.20. tert-butyl ((S)-1-((2S,4R)-4-hydroxy-2-(((S)-1-(4-(4-methylthiazol-5-yl)phenyl)ethyl) carbamoyl) pyrrolidin-1-yl)-3,3-dimethyl-1-oxobutan-2-yl) carbamate (P13) This solid material (P12), obtained as described above was dissolvedin 2 M HCl in EA (20 mL, 40 mmol), and the mixture was stirred atambient temperature for 12 h. The mixture was then concentrated, andthe residue was dried under vacuum to afford intermediate, which wasused in next step without further purification. HATU (8.50 g, 22.5 mmol,1.4 equiv) was added to a solution of this intermediate (5.80 g, 16.0mmol, 1.0 equiv), (S)-2-((tert-butoxycarbonyl) amino)-3,3- dimethylbutanoicacid (3.46 g, 16.0 mmol, 1.0 equiv), and DIPEA (8.8 mL,48.0 mmol, 3.0 equiv) in DMF (30 mL) at 0 C. The mixture was stirredat ambient temperature for 12 h when TLC showed that the reaction wascomplete. The reaction mixture was quenched with H2O (120 mL) andextracted with EtOAc (75 mL × 2). The combined organic layer waswashed with brine (100 mL) and dried over Na2SO4. The organic solutionwas filtered and concentrated. The residue was purified by silica gelflash column chromatography (DCM: MeOH 30:1) to afford the desiredcompound. | |
| With N-ethyl-N,N-diisopropylamine; N-[(dimethylamino)-3-oxo-1H-1,2,3-triazolo[4,5-b]pyridin-1-yl-methylene]-N-methylmethanaminium hexafluorophosphate In N,N-dimethyl-formamide at 20℃; | ||
| 84 % | With benzotriazol-1-ol; 1-ethyl-(3-(3-dimethylamino)propyl)-carbodiimide hydrochloride; N-ethyl-N,N-diisopropylamine In dichloromethane; N,N-dimethyl-formamide at -10 - 20℃; | |
| 15 % | With N-ethyl-N,N-diisopropylamine; N-[(dimethylamino)-3-oxo-1H-1,2,3-triazolo[4,5-b]pyridin-1-yl-methylene]-N-methylmethanaminium hexafluorophosphate In N,N-dimethyl-formamide at 16 - 20℃; | 6.f Synthesis of Int. 28 Into a 100 mL round-bottom flask were added (2S,4R)-4-hydroxy-N-[(1S)-1-[4-(4-methyl- 1,3-thiazol-5-yl)phenyl]ethyl]pyrrolidine-2-carboxamide (2.5 g, 7.543 mmol, 1.2 equiv), (2S)-2-[(tert-butoxycarbonyl)amino]-3,3-dimethylbutanoic acid (1.45 g, 6.286 mmol, 1 equiv), DMF (25 mL), HATU (3.59 g, 9.429 mmol, 1.5 equiv), and DIEA (2.44 g, 18.858 mmol, 3 equiv) and the mixture stirred for 1 h at 16 °C. The reaction was quenched with water at 16 °C. The residue was purified by reversed-phase flash chromatography on a C18 column eluting with a 5% to 60% gradient of MeCN in water (10 mmol/L NH4HCO3) over 40 min with UV (254 nm) detection. This resulted in tert-butyl N-[(2S)-1-[(2S,4R)-4-hydroxy-2-[(1S)- 1-[4-(4-methyl-1,3-thiazol-5-yl)phenyl]ethyl]carbamoyl}pyrrolidin-1-yl]-3,3-dimethyl-1- oxobutan-2-yl]carbamate (509.4 mg, 15% yield) as a white solid. LCMS: (ES, m/z): [M+H]+ = 545 (LCMS condition: Column: Shim-pack Scepter C18-120; Mobile phase A: Water/5 mM NH4HCO3; Mobile phase B: Acetonitrile; Flow rate: 1.5 mL/min; RT: 0.965 min) |

| Yield | Reaction Conditions | Operation in experiment |
|---|---|---|
| Multi-step reaction with 3 steps 1: hydrogenchloride / methanol; 1,4-dioxane / 12 h / 20 °C 2: N-ethyl-N,N-diisopropylamine; N-[(dimethylamino)-3-oxo-1H-1,2,3-triazolo[4,5-b]pyridin-1-yl-methylene]-N-methylmethanaminium hexafluorophosphate / N,N-dimethyl-formamide / 12 h / 0 - 20 °C / Inert atmosphere 3: hydrogenchloride / methanol; 1,4-dioxane / 12 h / 20 °C | ||
| Multi-step reaction with 3 steps 1: hydrogenchloride / 1,4-dioxane; methanol / 12 h / 20 °C 2: HATU; N-ethyl-N,N-diisopropylamine / N,N-dimethyl-formamide / 12 h / 0 - 20 °C / Inert atmosphere 3: hydrogenchloride / 1,4-dioxane; methanol / 12 h / 20 °C | ||
| Multi-step reaction with 3 steps 1: hydrogenchloride / dichloromethane; 1,4-dioxane / 1 h / 20 °C 2: triethylamine; N-[(dimethylamino)-3-oxo-1H-1,2,3-triazolo[4,5-b]pyridin-1-yl-methylene]-N-methylmethanaminium hexafluorophosphate / dichloromethane / 2 h / 20 °C 3: hydrogenchloride / dichloromethane; 1,4-dioxane / 1 h / 20 °C |
| Multi-step reaction with 3 steps 1: hydrogenchloride / 1,4-dioxane; dichloromethane / 1 h / 25 °C 2: N-[(dimethylamino)-3-oxo-1H-1,2,3-triazolo[4,5-b]pyridin-1-yl-methylene]-N-methylmethanaminium hexafluorophosphate; N-ethyl-N,N-diisopropylamine / tetrahydrofuran / 18 h / 20 °C 3: hydrogenchloride / 1,4-dioxane; dichloromethane / 1 h / 25 °C | ||
| Multi-step reaction with 3 steps 1.1: hydrogenchloride / methanol; 1,4-dioxane / 16 h / 25 °C 2.1: N-ethyl-N,N-diisopropylamine; N-[(dimethylamino)-3-oxo-1H-1,2,3-triazolo[4,5-b]pyridin-1-yl-methylene]-N-methylmethanaminium hexafluorophosphate / N,N-dimethyl-formamide / 15 min / 25 °C 2.2: 16 h / 25 °C 3.1: hydrogenchloride / 1,4-dioxane / 2 h / 25 °C | ||
| Multi-step reaction with 3 steps 1: trifluoroacetic acid / dichloromethane / 30 min / 20 °C 2: N-ethyl-N,N-diisopropylamine; HATU / N,N-dimethyl-formamide / 12 h / 20 °C 3: trifluoroacetic acid / dichloromethane / 30 min / 20 °C | ||
| Multi-step reaction with 3 steps 1: hydrogenchloride / ethyl acetate / 12 h / 20 °C 2: HATU; N-ethyl-N,N-diisopropylamine / N,N-dimethyl-formamide / 2 h / 0 - 20 °C / Inert atmosphere 3: hydrogenchloride / ethyl acetate / 12 h / 20 °C | ||
| Multi-step reaction with 3 steps 1: trifluoroacetic acid / dichloromethane / 30 min / 20 °C 2: N-ethyl-N,N-diisopropylamine; N-[(dimethylamino)-3-oxo-1H-1,2,3-triazolo[4,5-b]pyridin-1-yl-methylene]-N-methylmethanaminium hexafluorophosphate / N,N-dimethyl-formamide / 30 min / 20 °C 3: trifluoroacetic acid / dichloromethane / 30 min / 20 °C | ||
| Multi-step reaction with 3 steps 1: hydrogenchloride / 1,4-dioxane / 1 h / 16 °C 2: N-ethyl-N,N-diisopropylamine; N-[(dimethylamino)-3-oxo-1H-1,2,3-triazolo[4,5-b]pyridin-1-yl-methylene]-N-methylmethanaminium hexafluorophosphate / N,N-dimethyl-formamide / 1 h / 16 °C 3: hydrogenchloride / 1,4-dioxane / 1 h / 16 °C | ||
| Multi-step reaction with 3 steps 1: trifluoroacetic acid / dichloromethane / 1 h / 20 °C 2: N-ethyl-N,N-diisopropylamine; N-[(dimethylamino)-3-oxo-1H-1,2,3-triazolo[4,5-b]pyridin-1-yl-methylene]-N-methylmethanaminium hexafluorophosphate / N,N-dimethyl-formamide / 2 h / 20 °C 3: trifluoroacetic acid / dichloromethane / 1 h / 20 °C |

| Yield | Reaction Conditions | Operation in experiment |
|---|---|---|
| With hydrogenchloride In 1,4-dioxane; methanol at 20℃; for 12h; | ||
| With hydrogenchloride In 1,4-dioxane; methanol at 20℃; for 12h; | 1 tert- Butyl ((S)-l-((2S,4R)-4-hydroxy-2-(((S)-l-(4-(4-methylthiazol-5- yl)phenyl)ethyl)carb-amoyl)pyrrolidin- 1 -yl)-3 ,3 -dimethyl- 1 -oxobutan-2-yl)carbamate (57) This solid (56), obtained as described above was dissolved in 4N HC1 in dioxane (0327) (25 mL, 100 mmol) and MeOH (25 mL) and the mixture was stirred at ambient temperature for 12 h. The mixture was then concentrated and the residue was dried under vacuum to afford an intermediate, which was used in next step without further purification. UPLC-MS (ESL) calc, for C17H22N3O2S [M+l]+: 332.14, found 332.11. HATU (1.37 g, 3.6 mmol, 1.2 eq) was added to a solution of this intermediate (994 mg, 3.0 mmol, 1.0 eq), (S)-2-((/er/-butoxycarbonyl)amino)-3,3-dimethylbutanoic acid (694 mg, 3.0 mmol, 1.0 eq), and DIPEA (1.57 mL, 9.0 mmol, 3.0 eq) in DMF (10 mL) at 0 °C under N2. The mixture was stirred at ambient temperature for 12 h when TLC showed that the reaction was complete. The reaction mixture was quenched with H2O (100 mL) and extracted with EtOAc (75mLx2). The combined organic layer was washed with brine (100 mL) and dried over Na2S04. The organic solution was filtered and concentrated. The residue was purified by silica gel flash column chromatography with hexane :EtO Ac then DCM: MeOH to afford the desired compound (57) as a white solid (1.31 g, 80% yield). NMR (CDCI3, 400 MHz) d (ppm) 8.65 (s, 1H), 7.70 (d, J = 8.0 Hz, 1H), 7.35- 7.31 (m, 4H), 5.29 (d, J= 9.2 Hz, 1H), 5.06-5.01 (m, 1H), 4.67 (t, J= 8.0 Hz, 1H), 4.46- 4.44 (m, 1H), 4.22-4.19 (m, 1H), 3.91 (d, J = 17.2 Hz, 1H), 3.61-3.58 (m, 1H), 2.46 (s, 3H), 2.37-2.30 (m, 1H), 2.04-1.99 (m, 1H), 1.44 (d, J= 7.2 Hz, 3H), 1.35 (s, 9H), 0.96 (s, 9H); 13C NMR (CDCI3, 100 MHz) d (ppm) 172.22, 170.13, 156.15, 150.56, 148.21, 143.43, 131.74, 130.59, 129.49, 126.46, 80.18, 69.91, 58.86, 56.58, 48.74, 38.60, 36.02, 35.48, 28.34, 26.39, 22.17, 15.95; UPLC-MS (ESF) calc for C28H41N4O5S [M+l]+: 545.28, found 545.35. | |
| With hydrogenchloride In 1,4-dioxane; dichloromethane at 20℃; for 1h; |
| 420 mg | With hydrogenchloride In 1,4-dioxane; dichloromethane at 25℃; | E Step E: Add 1,4-dioxane solution (4 M, 4 ml) of hydrochloric acid with compound INT-5-4 (350 mg, 0.8 mmol) to dichloromethane (4 ml). The reaction mixture is stirred at 25 degrees Celsius for 1 h. LCMS monitoring shows that raw materials are gone. The reaction mixture was concentrated to remove the solvent to obtain 420 mg of INT-5-5. |
| 40 g | With hydrogenchloride In 1,4-dioxane at 25℃; | |
| With trifluoroacetic acid In dichloromethane at 20℃; | ||
| With hydrogenchloride In ethyl acetate at 20℃; | 4.1.20. tert-butyl ((S)-1-((2S,4R)-4-hydroxy-2-(((S)-1-(4-(4-methylthiazol-5-yl)phenyl)ethyl) carbamoyl) pyrrolidin-1-yl)-3,3-dimethyl-1-oxobutan-2-yl) carbamate (P13) This solid material (P12), obtained as described above was dissolvedin 2 M HCl in EA (20 mL, 40 mmol), and the mixture was stirred atambient temperature for 12 h. The mixture was then concentrated, andthe residue was dried under vacuum to afford intermediate, which wasused in next step without further purification. HATU (8.50 g, 22.5 mmol,1.4 equiv) was added to a solution of this intermediate (5.80 g, 16.0mmol, 1.0 equiv), (S)-2-((tert-butoxycarbonyl) amino)-3,3- dimethylbutanoicacid (3.46 g, 16.0 mmol, 1.0 equiv), and DIPEA (8.8 mL,48.0 mmol, 3.0 equiv) in DMF (30 mL) at 0 C. The mixture was stirredat ambient temperature for 12 h when TLC showed that the reaction wascomplete. The reaction mixture was quenched with H2O (120 mL) andextracted with EtOAc (75 mL × 2). The combined organic layer waswashed with brine (100 mL) and dried over Na2SO4. The organic solutionwas filtered and concentrated. The residue was purified by silica gelflash column chromatography (DCM: MeOH 30:1) to afford the desiredcompound. | |
| With hydrogenchloride In ethyl acetate at 20℃; | 4.1.20. tert-butyl ((S)-1-((2S,4R)-4-hydroxy-2-(((S)-1-(4-(4-methylthiazol-5-yl)phenyl)ethyl) carbamoyl) pyrrolidin-1-yl)-3,3-dimethyl-1-oxobutan-2-yl) carbamate (P13) This solid material (P12), obtained as described above was dissolvedin 2 M HCl in EA (20 mL, 40 mmol), and the mixture was stirred atambient temperature for 12 h. The mixture was then concentrated, andthe residue was dried under vacuum to afford intermediate, which wasused in next step without further purification. HATU (8.50 g, 22.5 mmol,1.4 equiv) was added to a solution of this intermediate (5.80 g, 16.0mmol, 1.0 equiv), (S)-2-((tert-butoxycarbonyl) amino)-3,3- dimethylbutanoicacid (3.46 g, 16.0 mmol, 1.0 equiv), and DIPEA (8.8 mL,48.0 mmol, 3.0 equiv) in DMF (30 mL) at 0 C. The mixture was stirredat ambient temperature for 12 h when TLC showed that the reaction wascomplete. The reaction mixture was quenched with H2O (120 mL) andextracted with EtOAc (75 mL × 2). The combined organic layer waswashed with brine (100 mL) and dried over Na2SO4. The organic solutionwas filtered and concentrated. The residue was purified by silica gelflash column chromatography (DCM: MeOH 30:1) to afford the desiredcompound. | |
| With hydrogenchloride In 1,4-dioxane; dichloromethane; water at 20℃; | Step 2: (2S,4R)-4-Hydroxy-N-((S)-1-(4-(4-methylthiazol-5-yl)phenyl)ethyl) pyrrolidine-2-carboxamide To a solution of tert-butyl (2S,4R)-4-hydroxy-2-[[(1S)-1-[4-(4-methylthiazol-5-yl)-phenyl]ethyl]carbamoyl]pyrrolidine-1-carboxylate (3.77 g, 8.74 mmol) in DCM (20 mL) was added 4M HCl-dioxane (20 mL, 80 mmol) and the reaction mixture was stirred at room temperature for 1 h. The solvent was evaporated under reduced pressure to afford the title compound. | |
| Multi-step reaction with 2 steps 1: hydrogenchloride / methanol / 2 h / 4 °C 2: sodium hydroxide / methanol; dichloromethane; water / 4 °C / pH 12.5 - 13 | ||
| With hydrogenchloride In 1,4-dioxane | 39.3 (2S,4R)-4-Hydroxy-N-((S)-1-(4-(4-methylthiazol-5-yl)phenyl)ethyl)pyrrolidine-2-carboxamide 39c Compound 1a (4.00 g, 9.27 mmol) was dissolved in hydrogen chloride/1,4-dioxane solution (50 mL, 4.0 M), stirred for 1 hour, and concentrated to obtain a crude product. The crude product was adjusted to neutrality with triethylamine and concentrated under reduced pressure to obtain the crude title compound 39c (5.06 g). The product was used directly in the next step without purification. | |
| With hydrogenchloride In 1,4-dioxane; dichloromethane at 20℃; | 47.2 Step 2: (2S,4R)-4-hydroxy-N-((S)-1-(4-(4-methylthiazol-5-yl) phenyl)ethyl) pyrrolidine-2-carboxamide To a stirred solution of tert-butyl (2S,4R)-4-hydroxy-2-[[(1S)-1-[4-(4-methylthiazol-5-yl)phenyl]ethyl]carbamoyl]pyrrolidine-1-carboxylate (3.77 g, 8.74 mmol, 1.00 eq.) in DCM (20 mL) was added HCl-dioxane (4.0 M, 20.0 mL). The resulting mixture was stirred at RT for 1 h and concentrated to afford the title compound. | |
| 16.8 g | With hydrogenchloride In 1,4-dioxane at 20℃; | Step 2: Synthesis of compound M29-2 M29-1 (22.0 g) was added to DCM (100 mL), and a hydrochloric acid dioxane solution (100 mL) was added under an ice-water bath. The reaction was reacted at room temperature for 60 minutes, and the reaction was completed by LCMS monitoring. The reaction solution was directly concentrated, and 1M NaOH solution was added until the pH of the reaction solution was 8. DCM (100 mL) was added to the reaction solution, and the organic phase was collected. The organic phase was washed once with saturated brine, dried over anhydrous sodium sulfate, filtered, and concentrated to obtain 16.8 g of a white solid, namely compound M29-2. |
| 98 % | With hydrogenchloride In 1,4-dioxane at 16℃; | 6.e Synthesis of Int. 27 Into a 100 mL round-bottom flask were added tert-butyl (2S,4R)-4-hydroxy-2-[(1S)-1-[4- (4-methyl-1,3-thiazol-5-yl)phenyl]ethyl]carbamoyl}pyrrolidine-1-carboxylate (2.8 g, 6.488 mmol, 1 equiv), 1,4-dioxane (10 mL) and HCl in 1,4-dioxane (0.79 mL, 4 M). The mixture was stirred for 1 h at 16 °C and was concentrated under reduced pressure. This afforded (2S,4R)-4-hydroxy-N-[(1S)-1-[4-(4-methyl-1,3-thiazol-5-yl)phenyl]ethyl]pyrrolidine-2- carboxamide (2.1 g, 98% yield) as a yellow green solid. LCMS: (ES, m/z): [M+H]+ = 332 (LCMS condition: Column: Shim-pack Scepter C18-120; Mobile phase A: Water/5 mM NH4HCO3; Mobile phase B: Acetonitrile; Flow rate: 1.5 mL/min) |
| With trifluoroacetic acid In dichloromethane at 20℃; for 1h; | TFA (2 mL) was added to a stirred solution of tert-butyl (2S,4R)-4-hydroxy-2-(((S)-1-(4-(4-methylthiazol-5- yl)phenyl)ethyl)carbamoyl)pyrrolidine-1-carboxylate (1 g, 2.3 mmol) in DCM (8 mL). The reaction mixture was stirred for 1h at room temperature. The reaction mixture was concentrated under reduced pressure to give a crude product that was purified by reverse phase FCC (column, C18 silica gel; mobile phase, ACN in Water (0.1% TFA), 10% to 50% gradient in 10 min; detector, UV 254 nm) affording the title compound (682 mg, 2.1 mmol) as a yellow solid. LCMS (ESI) m/z [M+H]+ = 332.1. | |
| 1.6 g | With trifluoroacetic acid In dichloromethane for 0.5h; | Synthesis of (2S,4R)-4-hydroxy-N-((S)-l-(4-(4-methylthiazol-5- yl)phenyl)ethyl)pyrrolidine-2 -carboxamide, 7. To a solution of 4 (2.00 g, 9.16 mmol, 1.0 eq), 5 (2.12 g, 9.16 mmol, 1.0 eq), HATU (5.22 g, 13.74 mmol, 1.5 eq) in DMF (50 mL) was added DIPEA (3.55 g, 27.48 mmol, 3.0 eq). The mixture was stirred for 30 minutes. The reaction was quenched by adding water. The mixture was extracted by ethyl acetate (80*2 mL) twice. Combined organic phases were washed by water twice. After dried by anhydrous Na2SO4and concentrated. The received residue was dissolved into DCM (40 mL), TFA (20 mL) was added, the mixture stirred for 30 minutes. After reaction, the solution was concentrated. The residue was purified by C-18 reversal column chromatography (elution solvents: CH3CN/H2O from 10% to 100%, 0.1%TFA) to get target product 7 (1.6 g, 53%) as a slight yellow solid. ESI MS m/z: 332.13 [M+H]+. |
| With trifluoroacetic acid In dichloromethane at 20℃; for 1h; | ||
| With hydrogenchloride In 1,4-dioxane; dichloromethane at 20℃; for 1h; | 59.2 Step 2: (2S,4R)-4-hydroxy-N-((S)-1-(4-(4-methylthiazol-5-yl)phenyl)ethyl) pyrrolidine-2-carboxamide To a solution of tert-butyl (2S,4R)-4-hydroxy-2-[(1S)-1-[4-(4-methylthiazol-5-yl)-phenyl]ethyl]carbamoyl]pyrrolidine-1-carboxylate (3.77 g, 8.74 mmol) in DCM (20 mL) wasadded 4M HCl-dioxane (20 mL, 80 mmol) and the reaction mixture was stirred at roomtemperature for 1 h. The solvent was evaporated under reduced pressure to afford the titlecompound. | |
| With hydrogenchloride In 1,4-dioxane; dichloromethane at 20℃; for 1h; | 2 Step 2: (2S,4R)-4-Hydroxy-N-((S)-1-(4-(4-methylthiazol-5-yl)phenyl)ethyl) pyrrolidine-2-carboxamide To a solution of tert-butyl (2S,4R)-4-hydroxy-2-[[(1S)-1-[4-(4-methylthiazol-5-yl)-phenyl]ethyl]carbamoyl]pyrrolidine-1-carboxylate (3.77 g, 8.74 mmol) in DCM (20 mL) wasadded 4M HCl-dioxane (20 mL, 80 mmol) and the reaction mixture was stirred at roomtemperature for 1 h. The solvent was evaporated under reduced pressure to afford the titlecompound. | |
| 682 mg | With trifluoroacetic acid In dichloromethane at 20℃; for 1h; | 6.2 Step 2. (2S,4R)-4-Hydroxy-N-((S)-1-(4-(4-methylthiazol-5-yl)phenyl)ethyl)pyrrolidine-2-carboxamide TFA (2 mL) was added to a stirred solution of tert-butyl (2S,4R)-4-hydroxy-2-(((S)-1-(4-(4-methylthiazol-5 yl)phenyl)ethyl) carbamoyl)pyrrolidine-1-carboxylate (1 g, 2.3 mmol) in DCM (8 mL). The reaction mixture was stirred for 1h at room temperature. The reaction mixture was concentrated under reduced pressure to give a crude product that was purified by reverse phase FCC (column, C18 silica gel; mobile phase, ACN in Water (0.1% TFA), 10% to 50% gradient in 10 min; detector, UV 254 nm) affording the title compound (682 mg, 2.1 mmol) as a yellow solid. LCMS (ESI) m/z [M+H]+ = 332.1. |

| Yield | Reaction Conditions | Operation in experiment |
|---|---|---|
| With triethylamine; N-[(dimethylamino)-3-oxo-1H-1,2,3-triazolo[4,5-b]pyridin-1-yl-methylene]-N-methylmethanaminium hexafluorophosphate In N,N-dimethyl-formamide at 0℃; for 2h; | ||
| With triethylamine; N-[(dimethylamino)-3-oxo-1H-1,2,3-triazolo[4,5-b]pyridin-1-yl-methylene]-N-methylmethanaminium hexafluorophosphate In N,N-dimethyl-formamide at 20℃; for 2h; | 59.3 Step 3: (9H-Fluoren-9-yl)methyl ((R)-1-((2S,4R)-4-hydroxy-2-(((S)-1-(4-(4-methylthiazol-5-yl)phenyl)ethyl)carbamoyl)pyrrolidin-1-yl)-3-methyl-1-oxo-3-(tritylthio)butan-2-yl)carbamate To a solution of (2R)-2-(9H-fluoren-9-ylmethoxy carbonylamino)-3-methyl-3-trityl-5 sulfanyl-butanoic acid (1576.6 mg, 2.57 mmol), (2S,4R)-4-hydroxy-N-[(1S)-1-[4-(4-methylthiazol-5-yl)phenyl]ethyl]pyrrolidine-2-carboxamide hydrochloride (1350 mg, 3.67mmmol) inDMF (10 mL) was added TEA (1.53 mL, 11.01 mmol), and HATU (2790.6 mg, 7.34 mmol), andthe mixture was stirred at room temperature for 2 h. The reaction mixture was diluted with water,extracted with EA. The combined organic layers were washed with brine, dried over anhydrous10 sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The residue waspurified by flash silica gel column chromatography to afford the title compound. | |
| With triethylamine; N-[(dimethylamino)-3-oxo-1H-1,2,3-triazolo[4,5-b]pyridin-1-yl-methylene]-N-methylmethanaminium hexafluorophosphate In N,N-dimethyl-formamide at 20℃; for 2h; | 3 Step 3: (9H-Fluoren-9-yl)methyl ((R)-1-((2S,4R)-4-hydroxy-2-(((S)-1-(4-(4-methylthiazol-5-yl)phenyl)ethyl)carbamoyl)pyrrolidin-1-yl)-3-methyl-1-oxo-3-(tritylthio)butan-2-yl)carbamate To a solution of (2R)-2-(9H-fluoren-9-ylmethoxycarbonylamino)-3-methyl-3-trityl-5 sulfanyl-butanoic acid (1576.6 mg, 2.57 mmol), (2S,4R)-4-hydroxy-N-[(1S)-1-[4-(4-methylthiazol-5-yl)phenyl]ethyl]pyrrolidine-2-carboxamide hydrochloride (1350 mg, 3.67mmmol) inDMF (10 mL) was added TEA (1.53 mL, 11.01 mmol), and HATU (2790.6 mg, 7.34 mmol), andthe mixture was stirred at room temperature for 2 h. The reaction mixture was diluted with water,extracted with EA. The combined organic layers were washed with brine, dried over anhydrous10 sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The residue waspurified by flash silica gel column chromatography to afford the title compound. |
| Yield | Reaction Conditions | Operation in experiment |
|---|---|---|
| Multi-step reaction with 4 steps 1: N-[(dimethylamino)-3-oxo-1H-1,2,3-triazolo[4,5-b]pyridin-1-yl-methylene]-N-methylmethanaminium hexafluorophosphate; N-ethyl-N,N-diisopropylamine / tetrahydrofuran / 18 h / 20 °C 2: hydrogenchloride / 1,4-dioxane; dichloromethane / 1 h / 25 °C 3: N-[(dimethylamino)-3-oxo-1H-1,2,3-triazolo[4,5-b]pyridin-1-yl-methylene]-N-methylmethanaminium hexafluorophosphate; N-ethyl-N,N-diisopropylamine / tetrahydrofuran / 18 h / 20 °C 4: hydrogenchloride / 1,4-dioxane; dichloromethane / 1 h / 25 °C |