3-马来酰亚胺基丙酸羟基琥珀酰亚胺酯

反应条件: temp:0.00; pressure:0.00; stir:; See reaction.notes.procedure_details.

实验步骤: Compound 14 (86 mg, 138 μmol) was dissolved in anhydrous acetonitrile (2 mL) and 3-(Maleimido)propionic acid N-hydroxysuccinimide ester (44 mg, 166 mmol) was added. The mixture was stirred overnight at room temperature, the solvent removed under reduced pressure and the residue purified over silica gel using chloroform/methanol (98/2) as eluant to give 86 mg (111 μmol, 81% yield) of a colorless oil. the solvent removed under reduced pressure the residue purified over silica gel

反应条件: temp:0.00; pressure:0.00; stir:; See reaction.notes.procedure_details.

实验步骤: 3-(2,5-Dioxo-2,5-dihydropyrrol-1-yl)propionic acid 2,5-dioxopyrrolidin-1-yl ester (787 mg, crude) was dissolved in DMF (15 ml). 8-Aminooctanoic acid (350 mg) and DIPEA (0.45 ml) was added. The mixture was stirred under nitrogen at room temperature over night. The mixture was concentrated. The residue was dissolved in EtOAc and extracted with 0.1 N HCl (2×). The phase separation was not easy. The organic layer was washed with brine (2×), dried (Na2SO4) and concentrated to give a white solid. LC-MS, M/z: 311.34 (M+1). The mixture was concentrated The residue was dissolved in EtOAc extracted with 0.1 N HCl (2×) The phase separation The organic layer was washed with brine (2×) dried (Na2SO4) concentrated to give a white solid

反应条件: temp:0.00; pressure:0.00; stir:; See reaction.notes.procedure_details.

实验步骤: Malimidopropionic acid (500 mg) was dissolved in dry THF (15 ml). TSTU (790 mg) and DIPEA (0.62 ml) was added. The mixture was stirred at room temperature under nitrogen over night. The yellow thick suspension was concentrated. The residue was dissolved in DCM and extracted with 0.1 N HCl (2×) and brine (1×). The organic layer was dried (Na2SO4) and concentrated to give a white solid. LC-MS, M/z: 267.26 (M+1). The yellow thick suspension was concentrated The residue was dissolved in DCM extracted with 0.1 N HCl (2×) and brine (1×) The organic layer was dried (Na2SO4) concentrated to give a white solid

反应条件: temp:0.00; pressure:0.00; stir:; See reaction.notes.procedure_details.

实验步骤: Malimidopropionic acid (500 mg) was dissolved in dry THF (15 ml). TSTU (790 mg) and DIPEA (0.62 ml) was added. The mixture was stirred at room temperature under nitrogen over night. The yellow thick suspension was concentrated. The residue was dissolved in DCM and extracted with 0.1 N HCl (2×) and brine (1×). The organic layer was dried (Na2SO4) and concentrated to give a white solid. LC-MS, M/z: 267.26 (M+1). The yellow thick suspension was concentrated The residue was dissolved in DCM extracted with 0.1 N HCl (2×) and brine (1×) The organic layer was dried (Na2SO4) concentrated to give a white solid

反应条件: temp:0.00; pressure:0.00; stir:; See reaction.notes.procedure_details.

实验步骤: 3-Maleimidopropionic acid N-hydroxysuccinimide ester (18) (4.0 g, 15.03 mmol, 1.0 equiv.) was dissolved in N,N-dimethylformamide (75 mL) and 3-aminopropyltriethoxysilane (17) (5.06 mL, 15.03 mmol, 1 equiv.) was added and the reaction mixture was allowed to stir for 22 hours. An aliquot (0.15 mL) was removed by syringe, concentrated in vacuo, and the resulting oil was analyzed by 1H NMR spectroscopy. The oil was consistent with desired product. The reaction mixture was then concentrated in vacuo at 50-55° C. and the residue obtained was dissolved in CH2Cl2 (75 mL), washed with water (3×50 mL) and brine (50 mL), filtered through granular Na2SO4, and concentrated under reduced pressure in vacuo to provide 6.65 g (>100% due to residual N,N-dimethylformamide) of the crude maleimide-silane (19) as a light yellow oil. Crude (19) was used without purification. 1H NMR (300 MHz, CDCl3) δ 6.65 (s, 2H), 6.00 (s, 1H), 3.77 (q, J=7 Hz, 6H), 3.70 (m, 2H), 3.17 (m, 2H), 2.46 (m, 2H), 1.56 (m, 2H), 1.18 (t, J=7 Hz, 9H), 0.57 (m, 2H). 13C NMR (75 MHz, CDCl3) δ 170.36, 169.28, 134.09, 58.35, 41.77, 34.56, 34.21, 23.20, 18.20, 7.65. FB-MS (pos): [M+H]+ 373 (373.2 calcd). An aliquot (0.15 mL) was removed by syringe concentrated in vacuo The reaction mixture was then concentrated in vacuo at 50-55° C. the residue obtained washed with water (3×50 mL) and brine (50 mL) filtered through granular Na2SO4 concentrated under reduced pressure in vacuo

反应条件: temp:0.00; pressure:0.00; stir:; See reaction.notes.procedure_details.

实验步骤: 3-Maleimidopropionic acid N-hydroxysuccinimide ester (18) (4.0 g, 15.03 mmol, 1.0 equiv.) was dissolved in N,N-dimethylformamide (75 mL) and 3-aminopropyltriethoxysilane (17) (5.06 mL, 15.03 mmol, 1 equiv.) was added and the reaction mixture was allowed to stir for 22 hours. An aliquot (0.15 mL) was removed by syringe, concentrated in vacuo, and the resulting oil was analyzed by 1H NMR spectroscopy. The oil was consistent with desired product. The reaction mixture was then concentrated in vacuo at 50-55° C. and the residue obtained was dissolved in CH2Cl2 (75 mL), washed with water (3×50 mL) and brine (50 mL), filtered through granular Na2SO4, and concentrated under reduced pressure in vacuo to provide 6.65 g (>100% due to residual N,N-dimethylformamide) of the crude maleimide-silane (19) as a light yellow oil. Crude (19) was used without purification. 1H NMR (300 MHz, CDCl3) δ 6.65 (s, 2H), 6.00 (s, 1H), 3.77 (q, J=7 Hz, 6H), 3.70 (m, 2H), 3.17 (m, 2H), 2.46 (m, 2H), 1.56 (m, 2H), 1.18 (t, J=7 Hz, 9H), 0.57 (m, 2H). 13C NMR (75 MHz, CDCl3) δ 170.36, 169.28, 134.09, 58.35, 41.77, 34.56, 34.21, 23.20, 18.20, 7.65. FB-MS (pos): [M+H]+ 373 (373.2 calcd). An aliquot (0.15 mL) was removed by syringe concentrated in vacuo The reaction mixture was then concentrated in vacuo at 50-55° C. the residue obtained washed with water (3×50 mL) and brine (50 mL) filtered through granular Na2SO4 concentrated under reduced pressure in vacuo

反应条件: temp:0.00; pressure:0.00; stir:; See reaction.notes.procedure_details.

实验步骤: A solution of β-alanine (1.8 g, 20 mmol) and maleic anhydride (2 g, 20 mmol) in DMF (30 mL) was stirred at room temperature for 3 h. After the solid was completely dissolved, N-hydroxysuccinimide (NHS; 2.3 g, 20 mmol) and dicyclohexylcarbodiimide (DCC; 4.6 g, 24 mmol) was added into the solution and the mixture was stirred at room temperature overnight. The solution was filtered and the precipitate was washed with water (100 ml) and dichloromethane (100 ml). The filtrate was collected and the organic layer was washed with 3×50 ml 5% NaHCO3 and one time with brine. The organic layer was dried with Na2SO4 and dichloromethane removed under reduced pressure to obtain a white solid (55%). This solid does not need further purification and can be directly used for next coupling step. 1H NMR (CDCl3, 400 MHz): δ 2.82 (s, 4H, two succinimido-CH2), 3.02 (t, J=7.0 Hz, 2H, COCH2), 3.94 (t, J=7.0 Hz, 2H, NCH2), 6.74 (s, 2H, two CH). After the solid was completely dissolved The solution was filtered the precipitate was washed with water (100 ml) and dichloromethane (100 ml) The filtrate was collected the organic layer was washed with 3×50 ml 5% NaHCO3 and one time with brine The organic layer was dried with Na2SO4 and dichloromethane removed under reduced pressure

反应条件: temp:0.00; pressure:0.00; stir:; See reaction.notes.procedure_details.

实验步骤: A solution of β-alanine (1.8 g, 20 mmol) and maleic anhydride (2 g, 20 mmol) in DMF (30 mL) was stirred at room temperature for 3 h. After the solid was completely dissolved, N-hydroxysuccinimide (NHS; 2.3 g, 20 mmol) and dicyclohexylcarbodiimide (DCC; 4.6 g, 24 mmol) was added into the solution and the mixture was stirred at room temperature overnight. The solution was filtered and the precipitate was washed with water (100 ml) and dichloromethane (100 ml). The filtrate was collected and the organic layer was washed with 3×50 ml 5% NaHCO3 and one time with brine. The organic layer was dried with Na2SO4 and dichloromethane removed under reduced pressure to obtain a white solid (55%). This solid does not need further purification and can be directly used for next coupling step. 1H NMR (CDCl3, 400 MHz): δ 2.82 (s, 4H, two succinimido-CH2), 3.02 (t, J=7.0 Hz, 2H, COCH2), 3.94 (t, J=7.0 Hz, 2H, NCH2), 6.74 (s, 2H, two CH). After the solid was completely dissolved The solution was filtered the precipitate was washed with water (100 ml) and dichloromethane (100 ml) The filtrate was collected the organic layer was washed with 3×50 ml 5% NaHCO3 and one time with brine The organic layer was dried with Na2SO4 and dichloromethane removed under reduced pressure

反应条件: temp:0.00; pressure:0.00; stir:; See reaction.notes.procedure_details.

实验步骤: 2,5-dioxopyrrolidin-1-yl 3-(2,5-dioxo-2,5-dihydro-1H-pyrrol-1-yl)propanoate (0.65 g, 2.43 mmol) was added to a solution of crude N-(3-(4-(4-((3-(2-(2-(3-aminopropoxy)ethoxy)ethoxy)propyl)amino)-benzoyl)phenoxy)propyl)-1-(5-(2-oxohexahydro-1H-thieno[3,4-d]imidazol-4-yl)pentanamido)-3,6,9,12-tetraoxapentadecan-15-amide (2 g, 2.11 mmol) in DCM (20 mL) and the reaction mixture was stirred for 2 hs at room temperature. Upon completion, reaction was then concentrated under reduced pressure, and purified by silica gel chromatography (DCM to DCM:MeOH, gradient) to yield 1.70 g (1.55 mmol, 73%) N-(3-(4-(4-((17-(2,5-dioxo-2,5-dihydro-1H-pyrrol-1-yl)-15-oxo-4,7,10-trioxa-14-azaheptadecyl)-amino)benzoyl)phenoxy)propyl)-1-(5-(2-oxohexahydro-1H-thieno[3,4-d]imidazol-4-yl)pentanamido)-3,6,9,12-tetraoxapentadecan-15-amide as yellow oil. Upon completion, reaction was then concentrated under reduced pressure purified by silica gel chromatography (DCM to DCM:MeOH, gradient)

反应条件: temp:0.00; pressure:0.00; stir:; See reaction.notes.procedure_details.

实验步骤: Compound 14 (86 mg, 138 μmol) was dissolved in anhydrous acetonitrile (2 mL) and 3-(Maleimido)propionic acid N-hydroxysuccinimide ester (44 mg, 166 mmol) was added. The mixture was stirred overnight at room temperature, the solvent removed under reduced pressure and the residue purified over silica gel using chloroform/methanol (98/2) as eluant to give 86 mg (111 μmol, 81% yield) of a colorless oil. the solvent removed under reduced pressure the residue purified over silica gel

反应条件: temp:0.00; pressure:0.00; stir:; See reaction.notes.procedure_details.

实验步骤: 3-(2,5-Dioxo-2,5-dihydropyrrol-1-yl)propionic acid 2,5-dioxopyrrolidin-1-yl ester (787 mg, crude) was dissolved in DMF (15 ml). 8-Aminooctanoic acid (350 mg) and DIPEA (0.45 ml) was added. The mixture was stirred under nitrogen at room temperature over night. The mixture was concentrated. The residue was dissolved in EtOAc and extracted with 0.1 N HCl (2×). The phase separation was not easy. The organic layer was washed with brine (2×), dried (Na2SO4) and concentrated to give a white solid. LC-MS, M/z: 311.34 (M+1). The mixture was concentrated The residue was dissolved in EtOAc extracted with 0.1 N HCl (2×) The phase separation The organic layer was washed with brine (2×) dried (Na2SO4) concentrated to give a white solid

反应条件: temp:0.00; pressure:0.00; stir:; See reaction.notes.procedure_details.

实验步骤: Malimidopropionic acid (500 mg) was dissolved in dry THF (15 ml). TSTU (790 mg) and DIPEA (0.62 ml) was added. The mixture was stirred at room temperature under nitrogen over night. The yellow thick suspension was concentrated. The residue was dissolved in DCM and extracted with 0.1 N HCl (2×) and brine (1×). The organic layer was dried (Na2SO4) and concentrated to give a white solid. LC-MS, M/z: 267.26 (M+1). The yellow thick suspension was concentrated The residue was dissolved in DCM extracted with 0.1 N HCl (2×) and brine (1×) The organic layer was dried (Na2SO4) concentrated to give a white solid