Structure of Val-Ala-PAB
CAS No.: 1343476-44-7
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| CAS No. : | 1343476-44-7 |
| Formula : | C15H23N3O3 |
| M.W : | 293.36 |
| SMILES Code : | CC(C)[C@H](N)C(N[C@@H](C)C(NC1=CC=C(C=C1)CO)=O)=O |
| English Name : | (S)-2-Amino-N-((S)-1-((4-(hydroxymethyl)phenyl)amino)-1-oxopropan-2-yl)-3-methylbutanamide |
| MDL No. : | MFCD32641649 |
| InChI Key : | NNWYWNRCBPYLML-GWCFXTLKSA-N |
| Pubchem ID : | 68223172 |
* 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 4-methyl-morpholine; O-(benzotriazol-1-yl)-N,N,N',N'-tetramethyluronium tetrafluoroborate In N,N-dimethyl-formamide at 20℃; for 72h; | 4.4A Coupling of the product (15) with linker, R-CO2H, was perfornned by activation with 1 equivalent of TBTU in the presence of 2 equivalents of NMM in DMF for 72 hours at room temperature to produce compound (16). |
| Yield | Reaction Conditions | Operation in experiment |
|---|---|---|
| Multi-step reaction with 2 steps 1: O-(benzotriazol-1-yl)-N,N,N',N'-tetramethyluronium tetrafluoroborate; 4-methyl-morpholine / N,N-dimethyl-formamide / 72 h / 20 °C 2: N,N-dimethyl-formamide |
| Yield | Reaction Conditions | Operation in experiment |
|---|---|---|
| Multi-step reaction with 3 steps 1: N,N-dimethyl-formamide / 6 h / 20 °C 2: N-ethyl-N,N-diisopropylamine / N,N-dimethyl-formamide / 5 h / 20 °C 3: N-ethyl-N,N-diisopropylamine; benzotriazol-1-ol / N,N-dimethyl-formamide / 20 °C | ||
| Multi-step reaction with 3 steps 1: N,N-dimethyl-formamide / 20 °C 2: N-ethyl-N,N-diisopropylamine / N,N-dimethyl-formamide / 5 h / 20 °C 3: N-ethyl-N,N-diisopropylamine; benzotriazol-1-ol / N,N-dimethyl-formamide / 20 °C | ||
| Multi-step reaction with 3 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 / N,N-dimethyl-formamide / 2 h / 20 °C 2: N-ethyl-N,N-diisopropylamine / N,N-dimethyl-formamide / 16 h / 20 °C 3: benzotriazol-1-ol; pyridine / dimethyl sulfoxide / 16 h / 20 °C |
| Yield | Reaction Conditions | Operation in experiment |
|---|---|---|
| Multi-step reaction with 2 steps 1: N-ethoxycarbonyl-2-ethoxy-1,2-dihydroquinoline / tetrahydrofuran / 72 h / 20 °C 2: tetrakis(triphenylphosphine) palladium(0); pyrrolidine / dichloromethane / 16.5 h / 20 °C / Inert atmosphere | ||
| Multi-step reaction with 2 steps 1: N-ethoxycarbonyl-2-ethoxy-1,2-dihydroquinoline / tetrahydrofuran / 40 h / 20 °C 2: tetrakis(triphenylphosphine) palladium(0); pyrrolidine / dichloromethane / 0.5 h / 20 °C / Inert atmosphere |
| Yield | Reaction Conditions | Operation in experiment |
|---|---|---|
| 74% | With N-ethoxycarbonyl-2-ethoxy-1,2-dihydroquinoline In tetrahydrofuran at 20℃; for 120h; | Compound 51 EEDQ (1.22 g, 4.93 mmol, 1.05 eq.) was added toa solution of amine dipeptide (50) (1.37 g, 4.69 mmol, 1 eq.) and m-dPeg2 acid (0.73 g, 4.93 mmol, 1.05 eq.) in dry THF (60 mE). The solution was stirred at room temperature for 5 days. The solvent was evaporated under reduced pressure. The residue was purified by flash column chromatography [100% chloroform to 95% chloroforml5% methanol in 1% increments] to give the product 51 as a white solid (1.46 g, 74%). Analytical Data: RT 2.22 mm; MS (ES) mlz (relative intensity) 446 ([M+Na], 80), 424 ([M+H], 70), MS (ES-) m/z (relative intensity) 422 ([M-H], 100). |
| Yield | Reaction Conditions | Operation in experiment |
|---|---|---|
| 76% | With triethylamine In N,N-dimethyl-formamide for 3.5h; | 1-4 Example 1-4: Synthesis of activated carbonate derivative 35 with cleavable linker To a solution of alkyne-PEG4-NHS ester (52 mg, 130 mitioI) in DMF (2 mL) were added 71 b (46 mg, 155 mitioI) and Et3N (54 pL, 39 mg, 390 mitioI). The resulting mixture was stirred for 3.5 h, diluted with DMF (0.5 mL) and concentrated. The residue was purified by silica chromatography (DCM 15% MeOH in DCM). The desired product was obtained as a white solid (57 mg, 76%). 1 H NMR (400 MHz, CDCI3) d (ppm) 8.66 (bs, 1 H), 7.73-6.65 (m, 2H), 7.33-7.25 (m, 2H), 7.20 (d, J = 7.9 Hz, 1 H), 7.16 (d, J = 6.8 Hz, 1 H), 4.68 (quintet, J = 7.5 Hz, 1 H), 4.62 (s, 2H), 4.22 (dd, J = 5.7 Hz, 6.8 Hz, 1 H), 4.18 (d, J = 2.4 Hz, 2H), 3.82 (dt, J = 9.7 Hz, 3.2 Hz, 1 H), 3.72-3.50 (m, 17H), 2.71-2.61 (m, 1 H), 2.51-2.41 (m, 1 H), 2.43 (t, J = 2.4 Hz, 1 H), 2.31-2.21 (m, 1 H), 1.44 (d, J = 7.2 Hz, 3H), 1 .01 (d, J = 6.9 Hz, 3H), 0.98 (d, J = 6.9 Hz, 3H). |
| Yield | Reaction Conditions | Operation in experiment |
|---|---|---|
| 76% | With N-ethyl-N,N-diisopropylamine In tetrahydrofuran; N,N-dimethyl-formamide at 20℃; for 0.5h; | 32.1 Step 1: 6-(11,12-Didehydrodibenzo[b,f]azocin-5(6H)-yl)-N-[(2S)-1-[(2S)-1-[4- (hydroxymethyl)phenyl]amino}-1-oxopropan-2-yl]amino}-3-methyl-1-oxobutan-2-yl]-6- oxohexanamide To a solution of (2S)-2-amino-N-[(1S)-2-[4-(hydroxymethyl)anilino]-1-methyl-2-oxo- ethyl]-3-methyl-butanamide (240 mg, 0.82 mmol) dissolved in DMF (2 mL) and THF (5.5 mL) was added N,N-diisopropylethylamine (0.39 mL, 2.24 mmol). The reaction mixture was cooled to 0 oC and 1-[6-(11,12-didehydrodibenzo[b,f]azocin-5(6H)-yl)-6- oxohexanoyl]oxy}pyrrolidine-2,5-dione (320 mg, 0.75 mmol) was added. The reaction mixture was warmed to rt and allowed to stir for 30 min. The reaction mixture was diluted with water and extracted with EtOAc (3x). The combined organic phases were washed with an aqueous saturated solution of sodium bicarbonate and brine, dried (Na2SO4), filtered and evaporated to dryness. The crude residue was purified by silica gel chromatography (50-100% EtOAc/DCM) to provide 6- (11,12-didehydrodibenzo[b,f]azocin-5(6H)-yl)-N-[(2S)-1-[(2S)-1-[4- (hydroxymethyl)phenyl]amino}-1-oxopropan-2-yl]amino}-3-methyl-1-oxobutan-2-yl]-6- oxohexanamide (347 mg, 76%). LCMS (AA): m/z = 609.4 (M+H). |
| Yield | Reaction Conditions | Operation in experiment |
|---|---|---|
| 163 mg | With benzotriazol-1-ol; 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 dichloromethane at 20℃; for 4h; | 13.5 [0352] Step 5: Preparation of (S)-2-(32-azido-5-oxo-3,9,12,15,18,21,24,27,30-nonoxy-6-diazapentatriacontamido)-N-((S)-1-((4-(hydroxymethyl)phenyl)amino)-1-oxopropan-2-yl)-3-methylbutyramide. At room temperature, O-benzotriazolyl-tetramethyluroniumhexafluorophosphate (160 mg, 0.42 mmol), 1-hydroxybenzotriazole (57 mg, 0.42 mmol), N,N-diisopropylethylamine (109 mg, 0.84 mmol) and 32-azido-5-oxo-3,9,12,15,18,21,24,27,30-nonoxy-6-azatricyclodecane-1-acid (156 mg, 0.28 mmol) were added to a solution of compound 30-5 (84 mg, 0.28 mmol) in dichloromethane (3 mL) and reacted for 4 hours under stirring. Purification was performed on silica gel column chromatography to obtain the title compound (163 mg). ESI-MS (m/z): 830.4 [M+H]+ |
| 7.8 g | With N-ethoxycarbonyl-2-ethoxy-1,2-dihydroquinoline In dichloromethane at 25℃; | 8.1 Step 1: (S)-2-(32-Azido-5-oxo-3,9,12,15,18,21,24,27,30-nonaoxa-6-azatritetradecamide )-N-((S)-1-((4-(hydroxymethyl)phenyl)amino)-1-oxopropan-2-yl)-3-methylbutylamine (8-3) Compound 8-2 (3.60g, 12.27mmol) was dissolved in dichloromethane (30mL), compound 8-1 (7.49g, 13.50mmol) and EEDQ (6.07g, 24.54mmol) were added, and reacted at 25°C for 4 hours. The reaction solution was concentrated under reduced pressure, and the concentrate was separated by reverse phase C18 column (70% acetonitrile/0.1% aqueous formic acid) to obtain the title compound 8-3 (7.80 g). |
| 7.8 g | With N-ethoxycarbonyl-2-ethoxy-1,2-dihydroquinoline In dichloromethane at 25℃; for 4h; | 8.1 Step 1: Synthesis of (S)-2-(3,2-azido-5-oxo-3,9,12,15,18,21,24,27,30-nonaoxa-6-azatriacontamide)-N-((S)-1-((4-(hydroxymethyl)phenyl)amino)-1-oxopropane-2-yl)-3-methylbutanamide (8-3) Compound 8-2 (3.60 g, 12.27 mmol) was dissolved in dichloromethane (30 mL), and compound 8-1 (7.49 g, 13.50 mmol) and EEDQ (6.07 g, 24.54 mmol) were added, and the mixture was reacted at 25°C for 4 hours. The reaction solution was concentrated under reduced pressure, and the concentrate was separated by a reverse phase C18 column (70% acetonitrile/0.1% formic acid aqueous solution) to obtain the title compound 8-3 (7.80 g). |
| Yield | Reaction Conditions | Operation in experiment |
|---|---|---|
| 1.64 g | With 1-ethyl-(3-(3-dimethylamino)propyl)-carbodiimide hydrochloride; N-ethyl-N,N-diisopropylamine In N,N-dimethyl-formamide at 20℃; for 16h; | 3.1 Step 1: (2S)-2-[3-[2-[2-[2-[2-[2-[2-[2-[2-[2-[2-[2-(2- azidoethoxy)ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethox y]ethoxy]propanoylamino]-N-[(1S)-2-[4-(hydroxymethyl)anilino]-1-methyl-2-oxo-ethyl]- 3-methyl-butanamide To a suspension of (2S)-2-amino-N-[(1S)-2-[4-(hydroxymethyl)anilino]-1-methyl-2- oxo-ethyl]-3-methyl-butanamide (900 mg, 3.07 µmol) in DMF (10 mL) were successively added a solution of 3-[2-[2-[2-[2-[2-[2-[2-[2-[2-[2-[2-(2- azidoethoxy)ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy ]propanoic acid (2.00 g, 3.07 mmol) in DMF (10 mL), EDC (650 mg, 3.38 mmol) as a powder and DIPEA (1.00 mL, 6.14 mmol). The reaction was stirred at room temperature for 16 h. The crude product was purified using C18 reverse phase prep-HPLC by direct deposit of the reaction mixture on the Xbridge column and using the NH4HCO3method to afford the desired product (1.64 g, 1.78 mmol). IR: (ν cm-1) 3600-3200, 3287, 2106, 1668, 1630, 1100.1H NMR (400 MHz, dmso-d6) δ ppm 9.82 (m, 1H), 8.14 (d, 1H), 7.87 (d, 1H), 7.54 (d, 2H), 7.23 (d, 2H), 5.08 (t, 1H), 4.43 (d, 2H), 4.39 (m, 1H), 4.20 (m, 1H), 3.65-3.44 (m, 48H), 3.39 (t, 2H), 2.5-2.3 (m, 2H), 1.97 (m, 1H), 1.31 (d, 3H), 0.87/0.84 (2d, 6H). HRMS (ESI) [M+H]+found: 919.5234 (δ = 3.4 ppm) |
| 1.64 g | With 1-ethyl-(3-(3-dimethylamino)propyl)-carbodiimide hydrochloride; N-ethyl-N,N-diisopropylamine In N,N-dimethyl-formamide at 20℃; for 16h; | 3.1 Step 1: (2S)-2-[3-[2-[2-[2-[2-[2-[2-[2-[2-[2-[2-[2-(2- azidoethoxy)ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethox y]ethoxy]propanoylamino]-N-[(1S)-2-[4-(hydroxymethyl)anilino]-1-methyl-2-oxo-ethyl]- 3-methyl-butanamide To a suspension of (2S)-2-amino-N-[(1S)-2-[4-(hydroxymethyl)anilino]-1-methyl-2- oxo-ethyl]-3-methyl-butanamide (900 mg, 3.07 µmol) in DMF (10 mL) were successively added a solution of 3-[2-[2-[2-[2-[2-[2-[2-[2-[2-[2-[2-(2- azidoethoxy)ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy ]propanoic acid (2.00 g, 3.07 mmol) in DMF (10 mL), EDC (650 mg, 3.38 mmol) as a powder and DIPEA (1.00 mL, 6.14 mmol). The reaction was stirred at room temperature for 16 h. The crude product was purified using C18 reverse phase prep-HPLC by direct deposit of the reaction mixture on the Xbridge column and using the NH4HCO3method to afford the desired product (1.64 g, 1.78 mmol). IR: (ν cm-1) 3600-3200, 3287, 2106, 1668, 1630, 1100.1H NMR (400 MHz, dmso-d6) δ ppm 9.82 (m, 1H), 8.14 (d, 1H), 7.87 (d, 1H), 7.54 (d, 2H), 7.23 (d, 2H), 5.08 (t, 1H), 4.43 (d, 2H), 4.39 (m, 1H), 4.20 (m, 1H), 3.65-3.44 (m, 48H), 3.39 (t, 2H), 2.5-2.3 (m, 2H), 1.97 (m, 1H), 1.31 (d, 3H), 0.87/0.84 (2d, 6H). HRMS (ESI) [M+H]+found: 919.5234 (δ = 3.4 ppm) |
| 1.64 g | With 1-ethyl-(3-(3-dimethylamino)propyl)-carbodiimide hydrochloride; N-ethyl-N,N-diisopropylamine; HATU In N,N-dimethyl-formamide at 20℃; | 1 Step 1: Synthesis of (2S)-2-[3-[2-[2-[2-[2-[2-[2-[2-[2-[2-[2-[2-(2- azidoethoxy)ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]eth oxy]ethoxy]propanoylamino]-N-[(1S)-2-[4-(hydroxymethyl)anilino]-1-methyl-2-oxo- ethyl]-3-methyl-butanamide To a solution of (2S)-2-amino-N-[(1S)-2-[4-(hydroxymethyl)anilino]-1-methyl-2-oxo- ethyl]-3-methyl-butanamide (0.9 g, 3.07 mmol; obtained according to Step 3 of the synthesis of L18-C3) and 3-[2-[2-[2-[2-[2-[2-[2-[2-[2-[2-[2-(2- azidoethoxy)ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy ]propanoic acid (purchased from Broadpharm, 2 g, 3.07 mmol) in DMF (20 mL) were successively added DIPEA (1 mL, 6.13 mmol), 3-(ethyliminomethyleneamino)propyl- dimethyl-ammonium; chloride (EDC) (0.65 g, 3.37 mmol) and [dimethylamino(triazolo[4,5- b]pyridin-3-yloxy)methylene]-dimethyl-ammonium; hexafluorophosphate (HATU) (1.28 g, 3.37 mmol). The mixture was stirred at room temperature overnight and purified by C18 reverse phase prep-HPLC by direct deposit of the reaction mixture on the Xbridge column and using the NH4HCO3 method to afford (2S)-2-[3-[2-[2-[2-[2-[2-[2-[2-[2-[2-[2-[2-(2- azidoethoxy)ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy ]propanoylamino]-N-[(1S)-2-[4-(hydroxymethyl)anilino]-1-methyl-2-oxo-ethyl]-3-methyl- butanamide (1.64 g, 1.81 mmol).1H NMR (400 MHz, dmso-d6): δ 9.82 (m, 1H), 8.14 (d, 1H), 7.87 (d, 1H), 7.54 (d, 2H), 7.23 (d, 2H), 5.08 (t, 1H), 4.43 (d, 2H), 4.39 (m, 1H), 4.2 (m, 1H), 3.65-3.44 (m, 48H), 3.39 (t, 2H), 2.50-2.30 (m, 2H), 1.97 (m, 1H), 1.31 (d, 3H), 0.87/0.84 (m, 6 H). IR Wavelength (cm-1): 3600-3200, 3287, 2106, 1668, 1630, 1100. HR-ESI+: m/z [M+H]+ = 919.5265 / 919.5234 (measured/theoretical). |
| 1.64 g | With 1-ethyl-(3-(3-dimethylamino)propyl)-carbodiimide hydrochloride; N-ethyl-N,N-diisopropylamine; HATU In N,N-dimethyl-formamide at 20℃; | 1 Step 1: Synthesis of (2S)-2-[3-[2-[2-[2-[2-[2-[2-[2-[2-[2-[2-[2-(2- azidoethoxy)ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]eth oxy]ethoxy]propanoylamino]-N-[(1S)-2-[4-(hydroxymethyl)anilino]-1-methyl-2-oxo- ethyl]-3-methyl-butanamide To a solution of (2S)-2-amino-N-[(1S)-2-[4-(hydroxymethyl)anilino]-1-methyl-2-oxo- ethyl]-3-methyl-butanamide (0.9 g, 3.07 mmol; obtained according to Step 3 of the synthesis of L18-C3) and 3-[2-[2-[2-[2-[2-[2-[2-[2-[2-[2-[2-(2- azidoethoxy)ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy ]propanoic acid (purchased from Broadpharm, 2 g, 3.07 mmol) in DMF (20 mL) were successively added DIPEA (1 mL, 6.13 mmol), 3-(ethyliminomethyleneamino)propyl- dimethyl-ammonium; chloride (EDC) (0.65 g, 3.37 mmol) and [dimethylamino(triazolo[4,5- b]pyridin-3-yloxy)methylene]-dimethyl-ammonium; hexafluorophosphate (HATU) (1.28 g, 3.37 mmol). The mixture was stirred at room temperature overnight and purified by C18 reverse phase prep-HPLC by direct deposit of the reaction mixture on the Xbridge column and using the NH4HCO3 method to afford (2S)-2-[3-[2-[2-[2-[2-[2-[2-[2-[2-[2-[2-[2-(2- azidoethoxy)ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy ]propanoylamino]-N-[(1S)-2-[4-(hydroxymethyl)anilino]-1-methyl-2-oxo-ethyl]-3-methyl- butanamide (1.64 g, 1.81 mmol).1H NMR (400 MHz, dmso-d6): δ 9.82 (m, 1H), 8.14 (d, 1H), 7.87 (d, 1H), 7.54 (d, 2H), 7.23 (d, 2H), 5.08 (t, 1H), 4.43 (d, 2H), 4.39 (m, 1H), 4.2 (m, 1H), 3.65-3.44 (m, 48H), 3.39 (t, 2H), 2.50-2.30 (m, 2H), 1.97 (m, 1H), 1.31 (d, 3H), 0.87/0.84 (m, 6 H). IR Wavelength (cm-1): 3600-3200, 3287, 2106, 1668, 1630, 1100. HR-ESI+: m/z [M+H]+ = 919.5265 / 919.5234 (measured/theoretical). |
| 1.64 g | With 1-ethyl-(3-(3-dimethylamino)propyl)-carbodiimide hydrochloride; N-ethyl-N,N-diisopropylamine In N,N-dimethyl-formamide at 20℃; | 3.1 Step 1: (2S)-2-[3-[2-[2-[2-[2-[2-[2-[2-[2-[2-[2-[2-(2- azidoethoxy)ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethox y]ethoxy]propanoylamino]-N-[(1S)-2-[4-(hydroxymethyl)anilino]-1-methyl-2-oxo-ethyl]- 3-methyl-butanamide To a suspension of (2S)-2-amino-N-[(1 S)-2-[4-(hydroxymethyl)anilino]-1 -methyl-2- oxo-ethyl]-3-methyl-butanamide (900 mg, 3.07 pmol) in DMF (10 mL) were successively added a solution of 3-[2-[2-[2-[2-[2-[2-[2-[2-[2-[2-[2-(2- azidoethoxy)ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy ]propanoic acid (2.00 g, 3.07 mmol) in DMF (10 mL), EDC (650 mg, 3.38 mmol) as a powder and DIPEA (1.00 mL, 6.14 mmol). The reaction was stirred at room temperature for 16 h.The crude product was purified using C18 reverse phase prep-HPLC by direct deposit of the reaction mixture on the Xbridge column and using the NH4HCO3 method to afford the desired product (1 .64 g, 1 .78 mmol). IR: (v cm1) 3600-3200, 3287, 2106, 1668, 1630, 1100.1H NMR (400 MHz, dmso-d6) 5 ppm 9.82 (m, 1 H), 8.14 (d, 1 H), 7.87 (d, 1 H), 7.54 (d, 2H), 7.23 (d, 2H), 5.08 (t, 1 H), 4.43 (d, 2H), 4.39 (m, 1 H), 4.20 (m, 1 H), 3.65-3.44 (m, 48H), 3.39 (t, 2H), 2.5-2.3 (m, 2H), 1 .97 (m, 1 H), 1 .31 (d, 3H), 0.87/0.84 (2d, 6H). HRMS (ESI) [M+H]+found: 919.5234 (5 = 3.4 ppm). |
| 1.64 g | With 1-ethyl-(3-(3-dimethylamino)propyl)-carbodiimide hydrochloride; N-ethyl-N,N-diisopropylamine In N,N-dimethyl-formamide at 20℃; | 3.F.1 Step 1: (2S)-2-[3-[2-[2-[2-[2-[2-[2-[2-[2-[2-[2-[2-(2- azidoethoxy)ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethox y]ethoxy]propanoylamino]-N-[(1S)-2-[4-(hydroxymethyl)anilino]-1-methyl-2-oxo-ethyl]- 3-methyl-butanamide To a suspension of (2S)-2-amino-N-[(1S)-2-[4-(hydroxymethyl)anilino]-1-methyl-2- oxo-ethyl]-3-methyl-butanamide (900 mg, 3.07 μmol) in DMF (10 mL) were successively added a solution of 3-[2-[2-[2-[2-[2-[2-[2-[2-[2-[2-[2-(2- azidoethoxy)ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy ]propanoic acid (2.00 g, 3.07 mmol) in DMF (10 mL), EDC (650 mg, 3.38 mmol) as a powder and DIPEA (1.00 mL, 6.14 mmol). The reaction was stirred at room temperature for 16 h. The crude product was purified using C18 reverse phase prep-HPLC by direct deposit of the reaction mixture on the Xbridge column and using the NH4HCO3 method to afford the desired product (1.64 g, 1.78 mmol). IR: (ν cm-1) 3600-3200, 3287, 2106, 1668, 1630, 1100. 1H NMR (400 MHz, dmso-d6) δ ppm 9.82 (m, 1H), 8.14 (d, 1H), 7.87 (d, 1H), 7.54 (d, 2H), 7.23 (d, 2H), 5.08 (t, 1H), 4.43 (d, 2H), 4.39 (m, 1H), 4.20 (m, 1H), 3.65-3.44 (m, 48H), 3.39 (t, 2H), 2.5-2.3 (m, 2H), 1.97 (m, 1H), 1.31 (d, 3H), 0.87/0.84 (2d, 6H). HRMS (ESI) [M+H]+ found: 919.5234 (δ = 3.4 ppm). |
| 1.64 g | With 1-ethyl-(3-(3-dimethylamino)propyl)-carbodiimide hydrochloride; N-ethyl-N,N-diisopropylamine In N,N-dimethyl-formamide at 20℃; | 3.1 Step 1: (2S)-2-[3-[2-[2-[2-[2-[2-[2-[2-[2-[2-[2-[2-(2- azidoethoxy)ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethox y]ethoxy]propanoylamino]-N-[(1S)-2-[4-(hydroxymethyl)anilino]-1-methyl-2-oxo-ethyl]- 3-methyl-butanamide To a suspension of (2S)-2-amino-N-[(1 S)-2-[4-(hydroxymethyl)anilino]-1 -methyl-2- oxo-ethyl]-3-methyl-butanamide (900 mg, 3.07 pmol) in DMF (10 mL) were successively added a solution of 3-[2-[2-[2-[2-[2-[2-[2-[2-[2-[2-[2-(2- azidoethoxy)ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy ]propanoic acid (2.00 g, 3.07 mmol) in DMF (10 mL), EDC (650 mg, 3.38 mmol) as a powder and DIPEA (1.00 mL, 6.14 mmol). The reaction was stirred at room temperature for 16 h.The crude product was purified using C18 reverse phase prep-HPLC by direct deposit of the reaction mixture on the Xbridge column and using the NH4HCO3 method to afford the desired product (1 .64 g, 1 .78 mmol). IR: (v cm1) 3600-3200, 3287, 2106, 1668, 1630, 1100.1H NMR (400 MHz, dmso-d6) 5 ppm 9.82 (m, 1 H), 8.14 (d, 1 H), 7.87 (d, 1 H), 7.54 (d, 2H), 7.23 (d, 2H), 5.08 (t, 1 H), 4.43 (d, 2H), 4.39 (m, 1 H), 4.20 (m, 1 H), 3.65-3.44 (m, 48H), 3.39 (t, 2H), 2.5-2.3 (m, 2H), 1 .97 (m, 1 H), 1 .31 (d, 3H), 0.87/0.84 (2d, 6H). HRMS (ESI) [M+H]+found: 919.5234 (5 = 3.4 ppm). |

| Yield | Reaction Conditions | Operation in experiment |
|---|---|---|
| 84.35% | Stage #1: 3-(2-(2-(3-(2,5-dioxo-2,5-dihydro-1H-pyrrol-1-yl)propanamido)ethoxy)ethoxy)propanoic acid With benzotriazol-1-ol; 1-ethyl-(3-(3-dimethylamino)propyl)-carbodiimide hydrochloride; N-ethyl-N,N-diisopropylamine In N,N-dimethyl-formamide at 20℃; for 2h; Inert atmosphere; Stage #2: (2S)-2-amino-N-[(1S)-2-[4-(hydroxymethyl)anilino]-1-methyl-2-oxoethyl]-3-methylbutanamide In N,N-dimethyl-formamide | 1.1 1) Preparation of Intermediates I-11a, I-12a, and I-13a Maleimide-diethylene glycol-carboxylic acid (3-(2-{2-[3-(2,5-Dioxo-2,5-dihydro-pyrrol-1-yl)-propionylamino]-ethoxy}-ethoxy)-propionic acid) (500.00 mg, 1.52 mmol) was added to a 100 mL eggplant-shaped flask and dissolved in 20 mL of anhydrous DMF. Under stirring at room temperature, EDCI (437.08 mg, 2.28 mmol), HOBt (247.27 mg, 1.83 mmol) and DIPEA (294.44 mg, 2.28 mmol) were added successively. The reaction was carried out at room temperature for 2 h under argon protection. Compound VA (Val-Ala) (445.91 mg, 1.52 mmol) was added to the reaction solution and monitored by TLC. After the reaction was completed, the solvent was removed by rotary evaporator and purified by column chromatography to obtain intermediate I-13a as a colorless oil (773.98 mg, 84.35% yield). |