Structure of 2316837-29-1
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| CAS No. : | 2316837-29-1 |
| Formula : | C15H23FN2O5 |
| M.W : | 330.35 |
| SMILES Code : | O=C(C1(CC1)F)N[C@@H](C(C)(C)C)C(N2[C@@H](C[C@H](C2)O)C(O)=O)=O |
| English Name : | (2S,4R)-1-((S)-2-(1-Fluorocyclopropane-1-carboxamido)-3,3-dimethylbutanoyl)-4-hydroxypyrrolidine-2-carboxylic acid |
| MDL No. : | MFCD34666431 |
| InChI Key : | QKWZXSRSTKMDNH-KXUCPTDWSA-N |
| Pubchem ID : | 153633266 |
* 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 20℃; for 0.5h; |
| Yield | Reaction Conditions | Operation in experiment |
|---|---|---|
| 94% | With water; lithium hydroxide In tetrahydrofuran at 20℃; | |
| 80% | With lithium hydroxide monohydrate In tetrahydrofuran; water at 0 - 20℃; for 18h; | (2S,4R)-1-((S)-2-(1-Fluorocyclopropane-1-carboxamido)-3,3-dimethylbutanoyl)-4-hydroxypyrrolidine-2-carboxylic Acid (E3LB-4(B)) To the solution of methyl (2S,4R)-1-((S)-2-(1-fluorocyclopropane-1-carboxamido)-3,3-dimethylbutanoyl)-4-hydroxypyrrolidine-2-carboxylate (BH) (0.65 g, 1.887 mmol) in THF (5.0 mL) at 0° C. was added the solution of lithium hydroxide monohydrate (0.792 g, 18.874 mmol) in water (2.5 mL). The resulting mixture was stirred at rt for 18 h. The organic solvent was removed under vacuo, the residue was diluted with ice-water (15 mL) and the pH was slowly adjusted to 2-3 with 2N HCl. The mixture was then extracted with EA (5 mL*4). The reunited organic layers were washed with brine (10 mL), dried over anhydrous Na2SO4 and then evaporated under reduced pressure to give a residue which was tritured with DEE, filtered off, and dried to give the titled compound as a white solid (0.497 g, 80% yield). 1H NMR (400 MHz, DMSO-d6): δ 12.55 (bs, 1H), 7.27 (d, J=6.9 Hz, 1H), 5.21 (d, J=3.7 Hz, 1H), 4.60 (d, J=9.1 Hz, 1H), 4.3-4.25 (m, 2H), 3.70-3.54 (m, 2H), 2.21-2.06 (m, 1H), 1.96-1.84 (m, 1H), 1.44-1.10 (m, 4H), 0.96 (s, 9H); 13C NMR (101 MHz, DMSO-d6): δ 173.56, 169.56, 168.53 (d, J=20.3 Hz), 78.55 (d, J=232.5 Hz), 69.28, 58.27, 56.88, 56.78, 37.69, 36.48, 26.57 (3C), 13.33 (dd, J=23.7, 10.3 Hz). |
| 71.4% | With lithium hydroxide monohydrate; water In methanol at 20℃; for 3h; |
| 70% | With lithium hydroxide monohydrate; water In tetrahydrofuran at 25℃; for 1h; | I.E Step E: (2S,4R)-1-((S)-2-(1-Fluorocyclopropane-1-carboxamido)-3,3-dimethylbutanoyl)-4-hydroxypyrrolidine-2-carboxylic acid To a solution of methyl (2S,4R)-1-((S)-2-(1-fluorocyclopropane-1-carboxamido)-3,3-dimethylbutanoyl)-4-hydroxypyrrolidine-2-carboxylate (5.0 g, 15.0 mmol) in THF (40.0 mL) and H2O (10.0 mL) was added lithium hydroxide (1.08 g, 45.3 mmol) and the reaction was stirred at 25 °C for 1 hour. The reaction mixture was adjusted to pH = 6 with 1 N HCl and extracted with DCM for 3 times. The combined organic layers were washed with brine, dried over anhydrous Na2SO4, filtered, and concentrated to afford (2S,4R)-1-((S)-2-(1- 111fluorocyclopropane-1-carboxamido)-3,3-dimethylbutanoyl)-4-hydroxypyrrolidine-2-carboxylic acid (5.30 g, 70%) as a white solid. MS(ES) [M+H]+331.1. |
| 720 mg | With lithium hydroxide monohydrate; water In tetrahydrofuran at 25℃; Inert atmosphere; | 1.2 (2S,4R)-1-((S)-2-(1-fluorocyclopropanecarboxamido)-3,3-dimethylbutanoyl)-4-hydroxypyrrolidine-2-carboxylic acid To a solution of (2S,4R)-methyl 1-((S)-2-(1-fluorocyclopropanecarboxamido)-3,3- dimethylbutanoyl)-4-hydroxypyrrolidine-2-carboxylate (1.00 g, 2.90 mmol) in THF (10 mL) was added LiOH.H2O (366 mg, 8.71 mmol) in H2O (5 mL). The mixture was stirred at 25 °C for 2 h. On completion, the pH of the reaction mixture was adjusted to 4~3 by addition 2 M HCl, then the mixture was extracted with EtOAc (10 mL x 10). The organic phase was separated, dried over Na2SO4, filtered and concentrated under reduced pressure to give the title compound (720 mg) as a white oil. |
| 720 mg | With lithium hydroxide monohydrate; water In tetrahydrofuran at 25℃; Inert atmosphere; | 1.2 (2S,4R)-1-((S)-2-(1-fluorocyclopropanecarboxamido)-3,3-dimethylbutanoyl)-4-hydroxypyrrolidine-2-carboxylic acid To a solution of (2S,4R)-methyl 1-((S)-2-(1-fluorocyclopropanecarboxamido)-3,3- dimethylbutanoyl)-4-hydroxypyrrolidine-2-carboxylate (1.00 g, 2.90 mmol) in THF (10 mL) was added LiOH.H2O (366 mg, 8.71 mmol) in H2O (5 mL). The mixture was stirred at 25 °C for 2 h. On completion, the pH of the reaction mixture was adjusted to 4~3 by addition 2 M HCl, then the mixture was extracted with EtOAc (10 mL x 10). The organic phase was separated, dried over Na2SO4, filtered and concentrated under reduced pressure to give the title compound (720 mg) as a white oil. |
| 70 % | With lithium hydroxide monohydrate In tetrahydrofuran; water at 20℃; | Step 4: Dissolve 8-c (1.5g, 4.3mmol) in THF (30mL) and water (10mL) and add LiOH ·H2O(900mg, 22.0mmol), 2 hours at room temperature, TLC monitoring reaction was completed. Concentrate to remove THF, add sodium hydroxide solid to adjust pH=12, add ethyl acetate (100mL) to extract, collect the aqueous phase, adjust the pH to 3-4 with hydrochloric acid solution (6mol/L) in the aqueous phase, extract with ethyl acetate (100mL×3), combine organic phase, anhydrous sodium sulfate drying, filtration, concentrate organic phase to obtain intermediate 8 (1.0g, white solid), yield: 70%. |
| 4.4 g | With water; sodium hydroxide In methanol for 3h; | Preparation of 6. To a solution of crude product 5 (5.00 g, 14.52 mmol, 1 equiv.) in MeOH/H2O (50 mL/5mL) was added NaOH (1.16 g, 28.28 mmol, 2.0 equiv.) The mixture was stirred for 3 h. MeOH was removed under rotatory evaporator. The aqueous phase was added TFA until the pH reach 4. The crude product aqueous solution was purified by C-18 reversal column (elution solvents: CH3CN/H2O from 10% to 100%, 0.1%TFA) to get pure product (4.4 g, 92%). ESI MS m/z: 331.15 [M+H]+. 1H NMR (400 MHz, DMSO) δ 12.81 (s, 1H), 7.26 (dd, J = 9.3, 3.0 Hz, 1H), 4.60 (dd, J = 9.4, 1.3 Hz, 1H), 4.44 - 4.27 (m, 2H), 3.66 (dd, J = 10.8, 3.9 Hz, 1H), 3.63 - 3.56 (m, 1H), 2.14 (ddt, J = 11.8, 7.9, 1.8 Hz, 1H), 1.91 (ddd, J = 13.3, 9.3, 4.5 Hz, 1H), 1.42 - 1.37 (m, 1H), 1.37 - 1.31 (m, 1H), 1.26 - 1.14 (m, 2H), 0.99 (s, 9H). |

| Yield | Reaction Conditions | Operation in experiment |
|---|---|---|
| 44% | 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℃; Inert atmosphere; | (2S,4R)-N-((S)-3-((6-(2-(2-(2,3-Difluoro-6-(2-morpholinothiazol-4-yl)phenoxy)acetamido)hexyl)amino)-1-(4-(4-methylthiazol-5-yl)phenyl)-3-oxopropyl)-1-((S)-2-(1-fluorocyclopropane-1-carboxamido)-3,3-dimethylbutanoyl)-4-hydroxypyrrolidine-2-carboxamide (Example 23) In an oven-dried round-bottom flask, under nitrogen atmosphere, to a stirred solution of (2S,4R)-1-((S)-2-(1-fluorocyclopropane-1-carboxamido)-3,3-dimethylbutanoyl)-4-hydroxypyrrolidine-2-carboxylic acid (E3LB-4(B)) (0.018 g, 0.054 mmol), (S)-3-amino-N-(6-(2-(2,3-difluoro-6-(2-morpholinothiazol-4-yl)phenoxy)acetamido)hexyl)-3-(4-(4-methylthiazol-5-yl)phenyl)propanamide hydrochloride (BP) (0.040 g, 0.054 mmol), and DIPEA (0.047 mL, 0.272 mmol) in dry DMF (0.7 mL) was added HATU (0.026 g, 0.068 mmol). Stirring was continued at rt overnight. The reaction mixture was poured in ice-water yielding a precipitate which was collected by filtration, dried, and purified by flash column chromatography on SiO2 (DCM/MeOH, 96:4 to 94:6) affording the titled compound as white solid (0.024 g, 44% yield). 1H NMR (400 MHz, CDCl3): δ 8.70 (s, 1H), 7.96 (d, J=8.1 Hz, 1H), 7.58 (t, J=6.5 Hz, 1H), 7.40 (s, 4H), 7.16-7.05 (m, 2H), 7.00 (dd, J=16.6, 8.9 Hz, 1H), 6.92 (s, 1H), 6.38 (bs, 1H), 5.39-5.32 (m, 1H), 4.69 (t, J=7.9 Hz, 1H), 4.66-4.57 (m, 3H), 4.54 (s, 1H), 4.01 (d, J=11.1 Hz, 1H), 3.89-3.82 (m, 4H), 3.74 (dd, J=11.0, 3.6 Hz, 1H), 3.57-3.51 (m, 4H), 3.32-3.24 (m, 2H), 3.23-3.14 (m, 2H), 2.87-2.71 (m, 2H), 2.53 (s, 3H), 2.32-2.17 (m, 2H), 1.52-1.40 (m, 4H), 1.39-1.25 (m, 9H), 1.09 (s, 9H); 13C NMR (101 MHz, CDCl3): δ 170.81, 170.74, 170.63, 170.01 (d, J=20.2 Hz), 169.95, 168.29, 150.86 (dd, J=251.6, 11.4 Hz), 150.36, 148.39, 146.08-145.96 (m), 144.58 (d, J=8.9 Hz), 144.20 (dd, J=247.36, 14.80 Hz), 140.55, 131.60, 131.02, 129.48 (2C), 126.82 (2C), 124.96-124.77 (m), 124.55 (dd, J=7.8, 4.0 Hz), 112.49 (d, J=17.1 Hz), 106.02, 78.32 (d, J=219.5 Hz), 72.28 (d, J=5.2 Hz), 70.20, 66.08 (2C), 59.33, 57.48, 56.67, 50.63, 48.63 (2C), 41.92, 39.06, 38.60, 37.08, 35.68, 29.28, 29.05, 26.48 (3C), 25.88, 25.82, 16.07, 13.68 (dd, J=10.2, 7.0 Hz) (2C). HRMS (ESI) m/z [M+H]+ calcd for C49H61F3N8O8S2 1011.40786. found 1011.40793. |
| Yield | Reaction Conditions | Operation in experiment |
|---|---|---|
| 83% | With lithium hydroxide monohydrate; water In methanol at 20 - 25℃; for 12h; | Step 3: To a solution of Intermediate 44-2 (3 g, 8.71 mmol) in MeOH (60 mL) and EbO (20 mL) was added LiOH.fbO (1.1 g, 26.13 mmol) at 25°C. The mixture was stirred at 20°C for 12 h and then concentrated to give a residue. The residue was dissolved in water (10 mL) and acidified to pH = 2 by the addition of cone. HC1. The resulting mixture was extracted with DCM : MeOH (5:1, 3 x 50 mL) and the combined organic layers were dried over Na2SO4 and filtered. The filtrate was concentrated to afford Intermediate 44 (2.4 g, 83% yield) as a white solid. LCMS (ESI) m/z 331.1 [M+H] +, |
| 83% | With lithium hydroxide monohydrate; water In methanol at 20 - 25℃; for 12h; | Step 3: To a solution of Intermediate 44-2 (3 g, 8.71 mmol) in MeOH (60 mL) and EbO (20 mL) was added LiOH.fbO (1.1 g, 26.13 mmol) at 25°C. The mixture was stirred at 20°C for 12 h and then concentrated to give a residue. The residue was dissolved in water (10 mL) and acidified to pH = 2 by the addition of cone. HC1. The resulting mixture was extracted with DCM : MeOH (5:1, 3 x 50 mL) and the combined organic layers were dried over Na2SO4 and filtered. The filtrate was concentrated to afford Intermediate 44 (2.4 g, 83% yield) as a white solid. LCMS (ESI) m/z 331.1 [M+H] +, |
| Yield | Reaction Conditions | Operation in experiment |
|---|---|---|
| 63% | With O-(1H-benzotriazol-1-yl)-N,N,N',N'-tetramethyluronium hexafluorophosphate; N-ethyl-N,N-diisopropylamine In N,N-dimethyl-formamide at 0 - 20℃; for 2h; | |
| 56% | 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℃; for 12h; | Step 4: To a solution of Intermediate 45-3 (2 g, 6.39 mmol) in DMF (20 mL) wa added DIPEA (5.28 mL, 31.97 mmol,), Intermediate 44 (2.53 g, 7.67 mmol) and HATU (2.92 g, 7.67 mmol) at 20°C. The mixture was stirred at rt for 12 h and then poured into H2O (20 ml) and extracted with EtOAc (3 x 50 mL). The combined organic layers were dried over Na2S04, filtered and concentrated. The crude was purified by HPLC (88:12 to 48:52 H2O (0.09% TFA): CH3CN) to obtain (S)-methyl 3-((2S,4R)-1-((S)-2-(l- fluorocyclopropanecarboxamido)-3,3-dimethylbutanoyl)-4-hydroxy pyrrolidine-2 - carboxamido)-3-(4-(4-methylthiazol-5-yl) phenyl)propanoate (Intermediate 45-4) (2.1 g, 56% yield) as a white solid. LCMS (ESI) m/z 589.4 [M+ |
| 56% | 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℃; for 12h; | Step 4: To a solution of Intermediate 45-3 (2 g, 6.39 mmol) in DMF (20 mL) wa added DIPEA (5.28 mL, 31.97 mmol,), Intermediate 44 (2.53 g, 7.67 mmol) and HATU (2.92 g, 7.67 mmol) at 20°C. The mixture was stirred at rt for 12 h and then poured into H2O (20 ml) and extracted with EtOAc (3 x 50 mL). The combined organic layers were dried over Na2S04, filtered and concentrated. The crude was purified by HPLC (88:12 to 48:52 H2O (0.09% TFA): CH3CN) to obtain (S)-methyl 3-((2S,4R)-1-((S)-2-(l- fluorocyclopropanecarboxamido)-3,3-dimethylbutanoyl)-4-hydroxy pyrrolidine-2 - carboxamido)-3-(4-(4-methylthiazol-5-yl) phenyl)propanoate (Intermediate 45-4) (2.1 g, 56% yield) as a white solid. LCMS (ESI) m/z 589.4 [M+ |
| With benzotriazol-1-ol; N-ethyl-N,N-diisopropylamine In N,N-dimethyl-formamide at 20℃; |

| Yield | Reaction Conditions | Operation in experiment |
|---|---|---|
| 81% | With 10% Pd/C; hydrogen In ethyl acetate at 20℃; for 16h; | |
| 76% | With 10% Pd/C; hydrogen In ethanol; ethyl acetate at 20℃; Inert atmosphere; | |
| With 10% Pd/C; hydrogen In ethanol |
| 75 % | With lithium hydroxide monohydrate In tetrahydrofuran; water at 0 - 23℃; |

| Yield | Reaction Conditions | Operation in experiment |
|---|---|---|
| 79 % | Stage #1: 9-(1-((3S,6S)-6-amino-3-methyl-6-(4-(4-methylthiazol-5-yl)phenyl)hexyl)piperidin-4-yl)-4-bromo-7-cyclopentylbenzo[4,5]imidazo[1,2-a]quinazolin-5(7H)-one bis-hydrochloride 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℃; Stage #2: (2S,4R)-1-((S)-2-(1-fluorocyclopropane-1-carboxamido)-3,3-dimethylbutanoyl)-4-hydroxypyrrolidine-2-carboxylic acid |
| Yield | Reaction Conditions | Operation in experiment |
|---|---|---|
| 68% | Stage #1: C17H21N5O2S With trifluoroacetic acid In dichloromethane for 0.5h; Stage #2: (2S,4R)-1-((S)-2-(1-fluorocyclopropane-1-carboxamido)-3,3-dimethylbutanoyl)-4-hydroxypyrrolidine-2-carboxylic acid 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 for 0.5h; | Synthesis of 5, To a solution of 2 (400 mg, 1.11 mmol, 1 eq) in DCM (10 mL) was added TFA (5 mL). the mixture was stirred for 30 minutes. After removing solvent and TFA, the residue was redissolved into DMF (10 mL), 4 (368 mg, 3.33 mmol, 1 eq.), HATU (637 mg, 1.67 mmol, 1.5 eq) and TEA (430 mg, 3.33 mmol, 3 eq) were added. The mixture was stirred for 30 minutes. The solution was purified by C-18 reversal column to get target compound 5 (430 mg, 68%) ESI MS m/z: 572.24 [M+H]+ |