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The BI-3802 was designed by Boehringer Ingelheim and could be obtained free of charge through the Boehringer Ingelheim open innovation portal opnMe.com, associated with its negative control.
Synonyms: 2-Maleimidoethylamine hydrochloride; N-(2-Aminoethyl)maleimide hydrochloride
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CAS No. : | 134272-64-3 |
Formula : | C6H9ClN2O2 |
M.W : | 176.60 |
SMILES Code : | O=C(C=C1)N(CCN)C1=O.[H]Cl |
Synonyms : |
2-Maleimidoethylamine hydrochloride; N-(2-Aminoethyl)maleimide hydrochloride
|
MDL No. : | MFCD09800491 |
InChI Key : | NJQOCRDPGFWEKA-UHFFFAOYSA-N |
Pubchem ID : | 22118207 |
GHS Pictogram: |
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Signal Word: | Warning |
Hazard Statements: | H315-H319 |
Precautionary Statements: | P305+P351+P338 |
* 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 |
---|---|---|
30.8% | With N-ethyl-N,N-diisopropylamine; In N,N-dimethyl-formamide; at 0 - 20℃; | To a solution of 4-azido-2-hydroxybenzoic acid N-hydroxysuccinimide ester (28 mg, 0.10 mmol) and N-(2-aminoethyl)maleimide hydrochloride (27 mg, 0.15 mmol) in DMF (250 muL) was added (i-Pr)2NEt (53 muL, 0.30 mmol) dropwise at 0 C. The reaction was allowed to warm room temperature, and stirred for 30 min. The reaction mixture was diluted with EtOAc, and then washed with 10% citric acid, H2O and brine. The organic layer was dried over Mg2SO4, and the solution was filtered, concentrated, and flash-chromatographed on silica gel to provide 1 (9.4 mg, 30.8%) as a pale yellow solid: mp >300 C; 1H NMR (CDCl3) delta 3.64 (m, 2H), 3.86 (t, J = 5.2 Hz , 2H) 6.53 (dd, J = 2.3 and 8.6 Hz, 1H) 6.62 (d, J = 2.3 Hz, 1H) 6.77 (s, 2H) 7.36 (d, J = 8.6 Hz, 1H); 13C NMR (CDCl3) delta 37.1, 39.9, 108.1, 110.1, 110.8, 127.2, 134.4, 146.0, 163.1, 169.8, 171.2. MS (ESI), calcd for C13H12N5O4 [M + H]+ 302.1, found 302.1 |
Yield | Reaction Conditions | Operation in experiment |
---|---|---|
Reaction of commercially available Boc-Gly-Gly-Gly-OH (compound 8) with Nhydroxyxuccinimide and EDC coupling agent affords compound 9. Reaction of compound 9 with 1-(2-aminoethyl)-maleimide HC1 in the presence of a base such as diisopropyl ethyl amine (DIPEA) followed by Boc deprotection with HC1 in methoxymethyl ether gives compound 10. Reaction of compound 10 with glutamic anhydride gives compound 11. Reaction of compound 11 with DM? using EDC as coupling agent will give the desired product compound 12. |
Yield | Reaction Conditions | Operation in experiment |
---|---|---|
In N,N-dimethyl acetamide; water; for 3h;Inert atmosphere; | A solution of 14.6 mM DM4 in DMA (2 mL, 29.2 mmol) was treated with 1:1 solution of saturated sodium bicarbonate to water (5%, 219 tl) followed by Sulfo-SPDB (29.7 mg, 0.073 mmol) under argon at room temperature with HPLC/MS monitoring. After 1 hour DM4 was consumed to give DM4-Sulfo-SPDB-DM4. Low Resolution MS calcd. (M1)= 1073.2; found (M-1)=1073.2.The reaction was treated with 1-(2-aminoethyl)-maleimide HC1 (12.90 mg, 0.073 mmol) under argon. After 3 hour desired product was formed. The material was purified on semi-prep HPLC C8 column using Water with 0.2% fomiic acid and 0.1% TEA and Acetonitrile. Low resolution MS, calcd. (M-1)=1098.34; found (M-1)=1098.2. |
Yield | Reaction Conditions | Operation in experiment |
---|---|---|
35% | With 1-ethyl-(3-(3-dimethylamino)propyl)-carbodiimide hydrochloride; In N,N-dimethyl-formamide; at 20℃; for 0.25h; | (DM 1 -GMB- Ala-Gly-Gly)2-Lys-b-Ala-OH (5.3 mg, 0.0022 mmol) was dissolved in anhydrous dimethylformamide (0.25 mL) to which was added EDC (2.0 mg, 0.014 mmol). After 2 min 2-amino-ethyl-maleimide HC1 salt (1 mg, 0.0057 mmol) was added and the solution was stirred at room temperature for 15 min. The reaction mixture was purified by HPLC on a XB-C18 21.2x150 mm, 5muiotaeta column with a flow rate of 21.2mL/min. eluting with deionized water containing 0.1% formic acid using a gradient of acetonitrile 5% for 4 min then a linear gradient of 5% - 95% over 17 min. Fractions containing desired product were combined, frozen and lyophilized to give 2 mg (35 % yield) of white solid. MS [M + Na]+ calcd. 2537.0; found 2537.3. |
Yield | Reaction Conditions | Operation in experiment |
---|---|---|
38% | With N-ethyl-N,N-diisopropylamine; In water; N,N-dimethyl-formamide; for 0.75h; | (SPDB-Ala-Gly-Gly)2-Lys-P-Ala-ONHS (22 mg, 0.020 mmol) was dissolved in anhydrous dimethyl formamide (400 mu) to which was added DM4 (30 mg, 0.038) with magnetic stirring. After 2 min deionized water (20 mu) was added then after 15 min diisopropylethyl amine (10 mu, mmol) and H2N-(CH2)2-Mal HC1 salt (3.7 mg, 0.022 mmol) were added. After an additional 30 min, sample was injected directly onto a 19 mm x 150 mm C18 HPLC column eluting with deionized water containing 0.2% formic acid with a 5-95% linear acetonitrile gradient. Flow rate was 20 mL/min. Fractions containing pure desired product were combined and frozen then lyophilized to give 19 mg (38% yield) of product as a white solid. MS [M + Na]+ calcd.2491.0; found 2491.3. |
Yield | Reaction Conditions | Operation in experiment |
---|---|---|
82% | With triethylamine; In tetrahydrofuran; at 20℃; for 16h; | To a solution of 1-(2-aminoethyl)-1H-pyrrol-2,5-dione 2,2,2-hydrochloride: (150mg, 0.849mmol) and TEA (350muL, 2.50mmol) in THF (4mL) was added (E)-perfluorophenyl 4-(phenyldiazenyl)benzoat 7 (666mg, 1.70mmol) in THF (4mL) dropwise. After 16h at rt, the solvent was evaporated, the resulting residue was dissolved in CHCl3 and washed with H2O (30mL), 1M HCl (30mL) and brine (30mL). The organic layer was dried over Na2SO4, filtered and concentrated. The crude product was purified by silica column (eluent EtOAc:hexane: 1:4?1:1; Rf:0.10?0.50) to yield the product as an orange solid (243mg; 82%). 1H NMR (500MHz, DMSO) delta 8.73 (t, J=6.0Hz, 1H), 7.96-7.88 (m, 6H), 7.67-7.56 (m, 3H), 7.01 (s, 2H), 3.62 (t, J=5.7Hz, 2H), 3.45 (dd, J=11.6, 5.9Hz, 2H); 13C NMR (126MHz, DMSO) delta 171.04, 165.76, 153.24, 151.89, 136.61, 134.51, 131.92, 129.49, 128.35, 122.67, 122.27, 37.71, 37.10; HPLC/MS (tr=13.92min; ESI) found: 349.1 (M+H)+, calc.: 349.1. |
Yield | Reaction Conditions | Operation in experiment |
---|---|---|
95.2% | With hydrogenchloride; In ethyl acetate; at 20℃; for 8h; | N-(2-((tert-Butoxycarbonyl)amino)ethyl)maleimide (750 mg, 3.12 mmol) was dissolved in 4 M HCl in ethyl acetate (20 mL) and stirred for 8 h at room temperature. Then, addition of diethyl ether at 0 C. provided the title compound as a white precipitate (524 mg, 2.97 mmol, 95.2% yield). 1H NMR (500 MHz, MeOD): delta 6.90 (s, 2H), 3.83-3.79 (m, 2H), 3.17-3.13 (m, 2H). |
Yield | Reaction Conditions | Operation in experiment |
---|---|---|
With trichlorophosphate; In tetrahydrofuran; at -78℃; for 2h; | N-2-ethyl-malimide hydrochloride salt (1.0 g, 5.66 mmol) in THF (50 ml) cooled at -78 C. was added phosphoryl trichloride (0.86 g, 5.66 mmol). After stirred at -78 C. for 2 h to form (2-(2,5-dioxo-2,5-dihydro-1H-pyrrol-1-yl)ethyl)phosphoramidic dichloride (2), 3-aminopropanoic acid (0.51 g, 5.70 mmol) in the mixture of THF/H2O (2:1, 30 ml) and triethylamine (1.0 g, 9.90 mmol) was added to the solution. The resulting mixture was stirred at RT for 3 h, concentrated under vacuum and purified on the SiO2 column eluted with H2O/CH3CN (1:201:10) to afford the title compound 4 (1.28 g, 78% yield). ESI MS m/z-C9H13N3O6P (M-H), cacld. 290.06. found 290.10. |
Yield | Reaction Conditions | Operation in experiment |
---|---|---|
Example 3 3-((Bis((2-(2,5-dioxo-2,5-dihydro-1H-pyrrol-1-yl)ethyl)amino)-phosphoryl)-amino)propanoic acid (8) N-2-ethyl-malimide hydrochloride salt (2.0 g, 11.32 mmol) in THF (100 ml) cooled at -78 C. was added phosphoryl trichloride (0.86 g, 5.66 mmol). After stirred at -78 C. for 1 h, the mixture was added triethylamine (1.0 g, 9.90 mmol) and the resulting solution was stirred at RT for 3 h to generate bis(2-(2,5-dioxo-2,5-dihydro-1H-pyrrol-1-yl)ethyl)-phosphoramidic chloride (7). Then 3-aminopropanoic acid (0.51 g, 5.70 mmol) in the mixture of THF/H2O (2:1, 30 ml) and triethylamine (1.51 g, 14.90 mmol) was added to the solution. The resulting mixture was stirred at 35 C. for 3 h, concentrated under vacuum and purified on the SiO2 column eluted with H2O/CH3CN (1:20?1:10) to afford the title compound 8 (1.47 g, 63% yield). ESI MS m/z-C15H19N5O7P (M-H), cacld. 412.11. found 412.20. |
Yield | Reaction Conditions | Operation in experiment |
---|---|---|
16% | To a suspension of the free thiol, 4a (88 mg, 0.105 mmol) and l-((2,5- dioxopyrrolidin-l-yl)oxy)-l-oxo-4-(pyridin-2-yldisulfanyl)butane-2-sulfonic acid (64.0 mg, 0.158 mmol) in anhydrous dichloromethane (2.10 mL) was added DIPEA (55.0 mu, 0.315 mmol) under nitrogen at room temperature. The mixture stirred for 16 hours and then l-(2- aminoethyl)-lH-pyrrole-2,5-dione hydrochloride (55.6 mg, 0.315 mmol), anhydrous dichloromethane (1.0 mL) and DIPEA (0.055 mL, 0.315 mmol) were added. The mixture stirred for an additional 5 hours at room temperature upon which the reaction was concentrated in vacuo. The resulting residue was purified by RP-HPLC (CI 8, CH3CN/H2O). Fractions containing desired product were frozen and lyophilized to give maleimide, compound D4 (20 mg, 16% yield) as a white solid. LCMS = 4.92 min (8 min method). MS (m/z): 1158.6 (M + 1)+. |
Yield | Reaction Conditions | Operation in experiment |
---|---|---|
80% | With N-ethyl-N,N-diisopropylamine; In dichloromethane; at 20℃; for 3h; | To a solution of the NHS ester, 6b (12.3 mg, 0.011 mmol) and N-(2- aminoethyl)maleimide hydrochloride (2.0 mg, 0.011 mmol) in anhydrous dichloromethane (0.3 mL) was added DIPEA (0.0022 mL, 0.013 mmol). The mixture was stirred at room temperature for 3 hours then it was stripped under reduced pressure. The residue was purified by semi-preparative reverse phase HPLC (CI 8 column, CH3CN/H2O). The fractions that contained pure product were combined, frozen and lyophilized to give the desired maleimide, compound D6 (10 mg, 80% yield). LCMS = 8.3 min (15 min method). MS (m/z): 1181.8 (M + D+). |
80% | With N-ethyl-N,N-diisopropylamine; In dichloromethane; at 20℃; for 3h; | Step 2: To a solution of the NHS ester, compound 6a (12.3 mg, 0.011 mmol) and N-(2- aminoethyl)maleimide hydrochloride (2.0 mg, 0.011 mmol) in anhydrous dichloromethane (0.3 niL) was added DIPEA (0.0022 niL, 0.013 mmol). The mixture was stirred at room temperature for 3 hours then it was stripped under reduced pressure. The residue was purified by semi-preparative reverse phase HPLC (CI 8 column, CH3CN/H2O). The fractions that contained pure product were combined, frozen and lyophilized to give the desired maleimide, compound D6 (10 mg, 80% yield). LCMS = 8.3 min (15 min method). MS (m/z): 1181.8 (M + 1)+. |
80% | With N-ethyl-N,N-diisopropylamine; In dichloromethane; at 20℃; for 3h; | To a solution of the NHS ester, compound 6a (12.3 mg, 0.011 mmol) and N-(2-aminoethyl)maleimide hydrochloride (2.0 mg, 0.011 mmol) in anhydrous dichloromethane (0.3 mL) was added DIPEA (0.0022 mL, 0.013 mmol). The mixture was stirred at room temperature for 3 hours then it was stripped under reduced pressure. The residue was purified by semi-preparative reverse phase HPLC (CI 8 column, CH3CN/H2O). The fractions that contained pure product were combined, frozen and lyophilized to give the desired maleimide, compound D6 (10 mg, 80% yield). LCMS = 8.3 min (15 min method). MS (m/z): 1181.8 (M + 1)+. |
80% | With N-ethyl-N,N-diisopropylamine; In dichloromethane; at 20℃; for 3h; | Step 2: To a solution of the NHS ester, compound 6a (12.3 mg, 0.011 mmol) and N-(2-aminoethyl)maleimide hydrochloride (2.0 mg, 0.011 mmol) in anhydrous dichloromethane (0.3 mL) was added DIPEA (0.0022 mL, 0.013 mmol). The mixture was stirred at room temperature for 3 hours then it was stripped under reduced pressure. The residue was purified by semi-preparative reverse phase HPLC (CI 8 column, CH3CN/H2O). The fractions that contained pure product were combined, frozen and lyophilized to give the desired maleimide, compound D6 (10 mg, 80% yield). LCMS = 8.3 min (15 min method). MS (m/z): 1181.8 (M + 1)+. |
80% | With N-ethyl-N,N-diisopropylamine; In dichloromethane; at 25℃; for 3h; | (0435) To a solution of the NHS ester, 2b (12.3 mg, 0.011 mmol) and N-(2-aminoethyl)maleimide hydrochloride (2.0 mg, 0.011 mmol) in anhydrous dichloromethane (0.3 mL) was added DIPEA (0.0022 mL, 0.013 mmol). The mixture was stirred at room temperature for 3 hours then it was stripped under reduced pressure. The residue was purified by semi-preparative reverse phase HPLC (C18 column, CH3CN/H2O). The fractions that contained pure product were combined, frozen and lyophilized to give the desired maleimide, compound D2 (10 mg, 80% yield). LCMS=8.3 min (15 min method). MS (m/z): 1181.8 (M+1)+. |
Yield | Reaction Conditions | Operation in experiment |
---|---|---|
58% | In dichloromethane; at 20℃; for 3.5h; | NHS ester 5a (8.2 mg, 7.6 muiotaetaomicron) and l-(2-aminoethyl)- lH-pyrrole-2,5-dione hydrochloride (2.2 mg, 0.011 mmol) were dissolved in anhydrous dichloromethane (305 ) at room temperature. DIPEA (2.66 mu, 0.015mmol) was added and the reaction and was stirred for 3.5 hours. The reaction mixture was concentrated and was purified by RPHPLC (CI 8 column, CH3CN/H2O, gradient, 35% to 55%). The desired product fractions were frozen and lyophilized to give maleimide, compound D5 as a solid white powder (5.3 mg, 58% yield). LCMS = 5.11 min (8 min method). MS (m/z): 1100.6 (M + 1)+. |
58% | With N-ethyl-N,N-diisopropylamine; In dichloromethane; at 20℃; for 3.5h; | NHS ester, compound 5a (8.2 mg, 7.6 muiotaetaomicron) and l-(2-aminoethyl)-lH-pyrrole-2,5-dione hydrochloride (2.2 mg, 0.011 mmol) were dissolved in anhydrous dichloromethane (305 mu) at room temperature. DIPEA (2.66 mu, 0.015mmol) was added and the reaction and was stirred for 3.5 hours. The reaction mixture was concentrated and was purified by RPHPLC (CI 8 column, CH3CN/H2O, gradient, 35% to 55%). The desired product fractions were frozen and lyophilized to give maleimide, compound D5 as a solid white powder (5.3 mg, 58% yield). LCMS = 5.11 min (8 min method). MS (m/z): 1100.6 (M + 1)+. |
58% | With N-ethyl-N,N-diisopropylamine; In dichloromethane; at 20℃; for 3.5h; | NHS ester, compound 5a (8.2 mg, 7.6 muiotaetaomicron) and l-(2-aminoethyl)-lH-pyrrole- 2,5-dione hydrochloride (2.2 mg, 0.011 mmol) were dissolved in anhydrous dichloromethane (305 mu) at room temperature. DIPEA (2.66 mu, 0.015mmol) was added and the reaction and was stirred for 3.5 hours. The reaction mixture was concentrated and was purified by RPHPLC (CI 8 column, CH3CN/H20, gradient, 35% to 55%). The desired product fractions were frozen and lyophilized to give maleimide, compound D5 as a solid white powder (5.3 mg, 58% yield). LCMS = 5.11 min (8 min method). MS (m z): 1100.6 (M + 1)+. |
58% | With N-ethyl-N,N-diisopropylamine; at 20℃; for 3.5h; | NHS ester, compound 5a (8.2 mg, 7.6 muiotaetaomicron) and l-(2-aminoethyl)-lH-pyrrole-2,5- dione hydrochloride (2.2 mg, 0.011 mmol) were dissolved in anhydrous dichloromethane (305 mu) at room temperature. DIPEA (2.66 mu, 0.015mmol) was added and the reaction and was stirred for 3.5 hours. The reaction mixture was concentrated and was purified by RPHPLC (CI 8 column, CH3CN/H20, gradient, 35% to 55%). The desired product fractions were frozen and lyophilized to give maleimide, compound D5 as a solid white powder (5.3 mg, 58% yield). LCMS = 5.11 min (8 min method). MS (m z): 1100.6 (M + 1)+. |
58% | With N-ethyl-N,N-diisopropylamine; In dichloromethane; at 25℃; for 3.5h; | (0432) NHS ester 1a (8.2 mg, 7.6 mumol) (prepared according to procedures described in US 2016/0082114, incorporated herein by reference) and <strong>[134272-64-3]1-(2-aminoethyl)-1H-pyrrole-2,5-dione hydrochloride</strong> (2.2 mg, 0.011 mmol) were dissolved in anhydrous dichloromethane (305 muL) at room temperature. DIPEA (2.66 muL, 0.015 mmol) was added and the reaction and was stirred for 3.5 hours. The reaction mixture was concentrated and was purified by RPHPLC (C18 column, CH3CN/H2O, gradient, 35% to 55%). The desired product fractions were frozen and lyophilized to give maleimide, compound D1 as a solid white powder (5.3 mg, 58% yield). LCMS=5.11 min (8 min method). MS (m/z): 1100.6 (M+1)+. |
Yield | Reaction Conditions | Operation in experiment |
---|---|---|
68% | With N-ethyl-N,N-diisopropylamine; In dichloromethane; at 20℃; for 4h;Inert atmosphere; | To a solution of NHS ester, 7a (5 mg, 4.82 muiotaetaomicron) and l-(2-aminoethyl)-lH-pyrrole- 2,5-dione hydrochloride (1.7 mg, 9.64 muiotaetaomicron) in anhydrous dichloromethane (200 ) was added DIPEA (1.512 mu, 8.68 muiotaetaomicron) under nitrogen. The mixture was stirred at room temperature for 4 hours and then concentrated in vacuo. The resulting residue was purified by RP-HPLC (CI 8, CH3CN/H2O). Fractions containing desired product were frozen and lyophilized to give maleimide, compound D7 (3.5 mg, 68% yield). LCMS = 4.61 min (15 min method). MS (m/z): 1062.8 (M + 1)+. |
68% | With N-ethyl-N,N-diisopropylamine; In dichloromethane; at 20℃; for 4h;Inert atmosphere; | To a solution of NHS ester, 7a (5 mg, 4.82 muiotaetaomicron) and l-(2-aminoethyl)-lH-pyrrole-2,5-dione hydrochloride (1.7 mg, 9.64 muiotaetaomicron) in anhydrous dichloromethane (200 mu) was added DIPEA (1.512 mu, 8.68 muiotaetaomicron) under nitrogen. The mixture was stirred at room temperature for 4 hours and then concentrated in vacuo. The resulting residue was purified by RP-HPLC (CI 8, CH3CN / H20). Fractions containing desired product were frozen and lyophilized to give maleimide, compound D7 (3.5 mg, 68% yield). LCMS = 4.61 min (15 min method). MS (m/z): 1062.8 (M + 1)+. |
68% | With N-ethyl-N,N-diisopropylamine; In dichloromethane; at 20℃; for 4h;Inert atmosphere; | To a solution of NHS ester, 7a (5 mg, 4.82 muetaiotaomicron) and l-(2-aminoethyl)-lH- pyrrole-2,5-dione hydrochloride (1.7 mg, 9.64 muiotaetaomicron) in anhydrous dichloromethane (200 mu) was added DIPEA (1.512 mu, 8.68 muiotaetaomicron) under nitrogen. The mixture was stirred at room temperature for 4 hours and then concentrated in vacuo. The resulting residue was purified by RP-HPLC (CI 8, CH3CN / H20). Fractions containing desired product were frozen and lyophilized to give maleimide, compound D7 (3.5 mg, 68% yield). LCMS = 4.61 min (15 min method). MS (m z): 1062.8 (M + 1)+. |
68% | With N-ethyl-N,N-diisopropylamine; In dichloromethane; at 20℃; for 4h;Inert atmosphere; | To a solution of NHS ester, 7a (5 mg, 4.82 muetaiotaomicron) and l-(2-aminoethyl)-lH-pyrrole- 2,5-dione hydrochloride (1.7 mg, 9.64 muiotaetaomicron) in anhydrous dichloromethane (200 mu) was added DIPEA (1.512 mu, 8.68 muiotaetaomicron) under nitrogen. The mixture was stirred at room temperature for 4 hours and then concentrated in vacuo. The resulting residue was purified by RP-HPLC (CI 8, CH3CN / H20). Fractions containing desired product were frozen and lyophilized to give maleimide, compound D7 (3.5 mg, 68% yield). LCMS = 4.61 min (15 min method). MS (m z): 1062.8 (M + 1)+. |
68% | With N-ethyl-N,N-diisopropylamine; In dichloromethane; at 25℃; for 4h;Inert atmosphere; | (0439) To a solution of NHS ester, 4a (5 mg, 4.82 mumol) and <strong>[134272-64-3]1-(2-aminoethyl)-1H-pyrrole-2,5-dione hydrochloride</strong> (1.7 mg, 9.64 mumol) in anhydrous dichloromethane (200 muL) was added DIPEA (1.512 muL, 8.68 mumol) under nitrogen. The mixture was stirred at room temperature for 4 hours and then concentrated in vacuo. The resulting residue was purified by RP-HPLC (C18, CH3CN/H2O). Fractions containing desired product were frozen and lyophilized to give maleimide, compound D4 (3.5 mg, 68% yield). LCMS=4.61 min (15 min method). MS (m/z): 1062.8 (M+1)+. |
Yield | Reaction Conditions | Operation in experiment |
---|---|---|
20 mg | With N-ethyl-N,N-diisopropylamine; In dichloromethane; at 20℃; for 5h;Inert atmosphere; | To a suspension of the free thiol, Dl (88 mg, 0.105 mmol) and l-((2,5-dioxopyrrolidin-l- yl)oxy)-l-oxo-4-(pyridin-2-yldisulfanyl)butane-2-sulfonic acid (sulfo-SPDB) (64.0 mg, 0.158 mmol) in anhydrous dichloromethane (2.10 mL) was added DIPEA (55.0 mu, 0.315 mmol) under nitrogen at room temperature. The mixture stirred for 16 hours and then l-(2- aminoethyl)-lH-pyrrole-2,5-dione hydrochloride (55.6 mg, 0.315 mmol), anhydrous dichloromethane (1.0 mL) and DIPEA (0.055 mL, 0.315 mmol) were added. The mixture stirred for an additional 5 hours at room temperature upon which the reaction was concentrated in vacuo. The resulting residue was purified by RP-HPLC (CI 8, CH3CN/H2O). Fractions containing desired product were frozen and lyophilized to give maleimide, D4 (20 mg, 16% yield) as a white solid. LCMS = 4.92 min (8 min method). MS (m/z): 1158.6 (M + D+. |
20 mg | With N-ethyl-N,N-diisopropylamine; In dichloromethane; at 20℃; for 5h; | To a suspension of the free thiol, Dl (88 mg, 0.105 mmol) and l-((2,5- dioxopyrrolidin-l-yl)oxy)-l-oxo-4-(pyridin-2-yldisulfanyl)butane-2-sulfonic acid (sulfo- SPDB) (64.0 mg, 0.158 mmol) in anhydrous dichloromethane (2.10 mL) was added DIPEA (55.0 mu, 0.315 mmol) under nitrogen at room temperature. The mixture stirred for 16 hours and then l-(2-aminoethyl)-lH-pyrrole-2,5-dione hydrochloride (55.6 mg, 0.315 mmol), anhydrous dichloromethane (1.0 mL) and DIPEA (0.055 mL, 0.315 mmol) were added. The mixture stirred for an additional 5 hours at room temperature upon which the reaction was concentrated in vacuo. The resulting residue was purified by RP-HPLC (CI 8, CH3CN/H2O). Fractions containing desired product were frozen and lyophilized to give maleimide, D4 (20 mg, 16% yield) as a white solid. LCMS = 4.92 min (8 min method). MS (m/z): 1158.6 (M + 1)+. |
Yield | Reaction Conditions | Operation in experiment |
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The synthesis of the compound tert-butyl (25S)-22-{(1R)-1-[1-benzyl-4-(2,5-difluorophenyl)-1H-imidazol-2-yl]-2,2-dimethylpropyl}-25-[(tert-butoxycarbonyl)amino]-1-(2,5-dioxo-2,5-dihydro-1H-pyrrol-1-yl)-4,21-dioxo-7,10,13,16-tetraoxa-19-thia-3,22-diazahexacosan-26-oate was carried out analogously to the synthesis of Intermediate C18. 37.4 mg (0.04 mmol) of (6S)-9-{(1R)-1-[1-benzyl-4-(2,5-difluorophenyl)-1H-imidazol-2-yl]-2,2-dimethylpropyl}-6-(tert-butoxycarbonyl)-2,2-dimethyl-4,10-dioxo-3,15,18,21,24-pentaoxa-12-thia-5,9-diazaheptacosan-27-oic acid. 9.2 mg (0.05 mmol) of <strong>[134272-64-3]1-(2-aminoethyl)-1H-pyrrole-2,5-dione hydrochloride</strong> (1:1). This gave 23.4 mg (49% of theory) of the compound tert-butyl (25S)-22-{(1R)-1-[1-benzyl-4-(2,5-difluorophenyl)-1H-imidazol-2-yl]-2,2-dimethylpropyl}-25-[(tert-butoxycarbonyl)amino]-1-(2,5-dioxo-2,5-dihydro-1H-pyrrol-1-yl)-4,21-dioxo-7,10,13,16-tetraoxa-19-thia-3,22-diazahexacosan-26-oate. LC-MS (Method 1): Rt=1.47 min; MS (ESIpos): m/z=1057 [M+H]+. |
Yield | Reaction Conditions | Operation in experiment |
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With N-ethyl-N,N-diisopropylamine; | tert-Butyl [22-{(1R)-1-[1-benzyl-4-(2,5-difluorophenyl)-1H-imidazol-2-yl]-2,2-dimethylpropyl}-1-(2,5-dioxo-2,5-dihydro-1H-pyrrol-1-yl)-4,21-dioxo-7,10,13,16-tetraoxa-19-thia-3,22-diazapentacosan-25-yl]carbamate 21.0 mg (0.03 mmol) of 9-{(1R)-1-[1-benzyl-4-(2,5-difluorophenyl)-1H-imidazol-2-yl]-2,2-dimethylpropyl}-2,2-dimethyl-4,10-dioxo-3,15,18,21,24-pentaoxa-12-thia-5,9-diazaheptacosan-27-oic acid (Intermediate C17) and 5.8 mg (0.0.3 mmol) of <strong>[134272-64-3]1-(2-aminoethyl)-1H-pyrrole-2,5-dione hydrochloride</strong> (1:1) were initially charged in 1.0 ml of acetonitrile, and 26.1 mg (0.20 mmol) of N,N-diisopropylethylamine and 20.9 mg (0.03 mmol) of T3P (50% in ethyl acetate) were added. The mixture was stirred at RT overnight. The reaction mixture was purified directly by preparative RP-HPLC (column: Reprosil 250*30; 10mu, flow rate: 50 ml/min, MeCN/water). The solvents were evaporated under reduced pressure and the residue was dried under high vacuum. This gave 19.7 mg (79% of theory) of the title compound. LC-MS (Method 1): Rt=1.42 min; MS (ESIpos): m/z=835 [M+H]+. |
Yield | Reaction Conditions | Operation in experiment |
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With N-ethyl-N,N-diisopropylamine; | Trifluoroacetic Acid/4-[{(1R)-1-[1-benzyl-4-(2,5-difluorophenyl)-1H-imidazol-2-yl]-2,2-dimethylpropyl}(glycoloyl)amino]methyl}-N-[2-(2,5-dioxo-2,5-dihydro-1H-pyrrol-1-yl)ethyl]pyrrolidine-3-carboxamide (1:1) 40.5 mg (0.06 mmol) of 4-[{(1R)-1-[1-benzyl-4-(2,5-difluorophenyl)-1H-imidazol-2-yl]-2,2-dimethylpropyl}(glycoloyl)amino]methyl}-1-(tert-butoxycarbonyl)pyrrolidine-3-carboxylic acid (Intermediate C25) and 14.5 mg (0.08 mmol) of <strong>[134272-64-3]1-(2-aminoethyl)-1H-pyrrole-2,5-dione hydrochloride</strong> (1:1) were initially charged in 1.0 ml of acetonitrile, and 64.4 mg (0.51 mmol) of N,N-diisopropylethylamine and 50.0 mg (0.08 mmol) of T3P were added and the mixture was stirred at RT for 16 h. The same amount of <strong>[134272-64-3]1-(2-aminoethyl)-1H-pyrrole-2,5-dione hydrochloride</strong> (1:1), N,N-diisopropylethylamine and T3P were added again, and the mixture was stirred at RT for a further 4 h. The reaction mixture was purified directly by preparative RP-HPLC (column: Reprosil 125*30; 10mu, flow rate: 50 ml/min, MeCN/water). The solvents were evaporated under reduced pressure and the residue was dried under high vacuum. This gave 7.2 mg (15% of theory) of the compound tert-butyl 3-[{(1R)-1-[1-benzyl-4-(2,5-difluorophenyl)-1H-imidazol-2-yl]-2,2-dimethylpropyl}(glycoloyl)amino]methyl}-4-[2-(2,5-dioxo-2,5-dihydro-1H-pyrrol-1-yl)ethyl]carbamoyl}pyrrolidine-1-carboxylate. LC-MS (Method 1): Rt=1.30 min; MS (ESIpos): m/z=763 [M+H]+. |
Yield | Reaction Conditions | Operation in experiment |
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16% | (0436) (0437) To a suspension of the free thiol, 3a (88 mg, 0.105 mmol) and 1-((2,5-dioxopyrrolidin-1-yl)oxy)-1-oxo-4-(pyridin-2-yldisulfanyl)butane-2-sulfonic acid (64.0 mg, 0.158 mmol) in anhydrous dichloromethane (2.10 mL) was added DIPEA (55.0 muL, 0.315 mmol) under nitrogen at room temperature. The mixture stirred for 16 hours and then <strong>[134272-64-3]1-(2-aminoethyl)-1H-pyrrole-2,5-dione hydrochloride</strong> (55.6 mg, 0.315 mmol), anhydrous dichloromethane (1.0 mL) and DIPEA (0.055 mL, 0.315 mmol) were added. The mixture stirred for an additional 5 hours at room temperature upon which the reaction was concentrated in vacuo. The resulting residue was purified by RP-HPLC (C18, CH3CN/H2O). Fractions containing desired product were frozen and lyophilized to give maleimide, compound D3 (20 mg, 16% yield) as a white solid. LCMS=4.92 min (8 min method). MS (m/z): 1158.6 (M+1)+. | |
16% | To a suspension of the free thiol, Dl (88 mg, 0.105 mmol) and l-((2,5- dioxopyrrolidin-l-yl)oxy)-l-oxo-4-(pyridin-2-yldisulfanyl)butane-2-sulfonic acid (sulfo- SPDB) (64.0 mg, 0.158 mmol) in anhydrous dichloromethane (2.10 mL) was added DIPEA (55.0 mu, 0.315 mmol) under nitrogen at room temperature. The mixture stirred for 16 hours and then l-(2-aminoethyl)-lH-pyrrole-2,5-dione hydrochloride (55.6 mg, 0.315 mmol), anhydrous dichloromethane (1.0 mL) and DIPEA (0.055 mL, 0.315 mmol) were added. The mixture stirred for an additional 5 hours at room temperature upon which the reaction was concentrated in vacuo. The resulting residue was purified by RP-HPLC (CI 8, CH3CN/H20). Fractions containing desired product were frozen and lyophilized to give maleimide, D4 (20 mg, 16% yield) as a white solid. LCMS = 4.92 min (8 min method). MS (m/z): 1158.6 (M + 1)+. |