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CAS No. : | 159857-80-4 | MDL No. : | MFCD26142966 |
Formula : | C28H40N6O7 | Boiling Point : | - |
Linear Structure Formula : | - | InChI Key : | AMKBTTRWLGVRER-OFVILXPXSA-N |
M.W : | 572.65 | Pubchem ID : | 57544445 |
Synonyms : |
MC-Val-Cit-PAB-OH
|
Chemical Name : | 6-(2,5-Dioxo-2,5-dihydro-1H-pyrrol-1-yl)-N-((S)-1-(((S)-1-((4-(hydroxymethyl)phenyl)amino)-1-oxo-5-ureidopentan-2-yl)amino)-3-methyl-1-oxobutan-2-yl)hexanamide |
Signal Word: | Warning | Class: | N/A |
Precautionary Statements: | P261-P305+P351+P338 | UN#: | N/A |
Hazard Statements: | H302-H315-H319-H335 | Packing Group: | N/A |
GHS Pictogram: |
* 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 |
---|---|---|
57% | With N-ethyl-N,N-diisopropylamine In dichloromethane; N,N-dimethyl-formamide at 23℃; for 20 h; | To a solution of Compound 11 (500 mg, 0.87 mmol) and bis(4-nitrophenyl) carbonate (bis-PNP) (2.64 g, 8.72 mmol) in DCM:DMF (8:2, 25 mL) was added DIPEA (0.45 mL, 2.61 mmol). The reaction mixture was stirred for 20 h at 23 °C and poured onto a silica gel column (DCM:CH3OH, from 50:1 to 10:1 ) to afford pure target Compound 9 (364 mg, 57percent). Rf= 0.40 (CH2CI2:CH3OH, 9:1 ).1H NMR (400 MHz, CDCI3/CD3OD): δ 9.45 (s, 1 H), 8.23 (d, J = 8.3 Hz, 2H), 7.59 (d, J = 8.5 Hz, 2H), 7.35 (d, J = 8.3 Hz, 2H), 7.34 (d, J = 8.5 Hz, 2H), 6.65 (s, 2H), 5.20 (s, 2H), 4.56 (dt, J = 10.5, 5.4 Hz, 1 H), 4.15 (d, J = 7.2 Hz, 1 H), 3.46 (dd, J = 8.0, 6.4 Hz, 2H), 3.16-2.89 (m, 2H), 2.21 (dd, J = 8.3, 6.6 Hz, 2H), 2.06-1 .97 (m, 1 H), 1.90-1.83 (m, 1 H), 1.73-1.46 (m, 7H), 1.34- 1.20 (m, 2H), 0.91 (d, J = 6.7 Hz, 3H), 0.90 (d, = 6.7 Hz, 3H).13C NMR (125 MHz, CDCI3/CD3OD) δ 174.4, 172.4, 171.1 , 170.6, 160.5, 155.5, 152.5, 145.3, 138.7, 134.1 , 129.9, 129.5, 125.2, 121.8, 120.0, 70.6, 59.0, 53.2, 37.5, 35.8, 30.6, 29.6, 29.3, 28.1 , 26.2, 26.2, 25.1 , 19.1 , 18.1. ESI-MS m/z: Calcd. for CasH^N O^: 737.3. Found: 738.3 (M+H)+. |
57% | With N-ethyl-N,N-diisopropylamine In N,N-dimethyl-formamide at 0 - 20℃; for 15 h; Inert atmosphere | Preparation Example 1-2: Preparation of Compound (II-3) (0054) (0055) Under an argon stream, 6-(2,5-dioxo-2,5-dihydro-1H-pyrrol-1-yl)-N-((S)-1-(((S)-1-((4-(hydroxymethyl)phenyl)amino)-1-oxo-5-ureidopentan-2-yl)amino)-3-methyl-1-oxobutan-2-yl)hexanamide(5.0 g, 8.7 mmol, Dubowchik et al., Bioconjugate Chem., 2002, 13 (4), pp 855-869) was dissolved in 60 mL of anhydrous dimethylformamide, added with N, N-diisopropylethylamine (3.0 mL, 17.4 mmol), and then cooled to 0°C. To the mixture was added at once bis(4-nitrophenyl) carbonate (7.94 g, 26.1 mmol), followed by stirring at room temperature for 15 hrs. After completion of the reaction, the reaction mixture was concentrated in a high vacuum, and the concentrate was purified by silica gel column chromatography to obtain 4-((S)-2-((S)-2-(6-(2 ,5-dioxo-2,5-dihydro-1H-pyrrol-1-yl)hexanamido)-3-methylbutanamido)-5-ureidopentanamido)benzyl (4-nitrophenyl) carbonate (3.65 g, 57 percent) as a pale yellow solid. (0056) 1H NMR (400 MHz, DMSO-d6) δ 0.83 (d, J = 6.8 Hz, 3 H), 0.86 (d, J = 6.8 Hz, 3 H), 1.09 (t, J = 7.2 Hz, 1 H), 1.19 (m, 2 H), 1.34 - 1.76 (m, 7 H), 1.96 (m, 1 H), 2.15 (m, 2 H), 2.99 (m, 2 H), 3.37 (m, 2 H), 4.19 (t, J= 7.8 Hz, 1 H), 4.39 (m, 1 H), 5.24 (s, 2 H), 5.41 (s, 2 H), 5.97 (brt, J= 5.6 Hz, 1 H), 7.00 (s, 2H), 7.41 (d, J= 8.4 Hz, 2 H), 7.57 (d, J= 7.2 Hz, 2 H), 7.65 (d, J= 8.4 Hz, 2 H), 7.80 (d, J= 8.4 Hz, 1 H), 8.09 (d, J = 7.2 Hz, 1 H), 8.31 (d, J = 7.2 Hz, 2 H), 10.05 (brs, 1 H) (0057) Under an argon stream, 4-((S)-2-((S)-2-(6-(2, 5-dioxo-2,5-dihydro-1H-pyrrol-1-yl)hexanamido)-3-methylbutanamido)-5-ureidopentanamido)benzyl (4-nitrophenyl) carbonate (3.65 g, 4.95 mmol) was dissolved in 90 mL of anhydrous dimethylformamide, and stirred, together with t-butyl (2-aminoethyl)(methyl)carbamate (0.86 g, 4.95 mmol), at 20 - 25°C for 2 hrs. After completion of the reaction, the reaction mixture was completely concentrated in a high vaccum, and the concentrate was purified by silica gel column chromatography to obtain an amino-protected derivative of compound (II-3) (3.8 g, 99percent). (0058) LC-MS m/z: 773.5 [M+H]+ (0059) To a solution of the amino-protected derivative of compound (II-3) (146 mg, 0.186 mmol) in 5 mL of dichloromethane was dropwise added 2 mL of trifluoroacetic acid, followed by stirring at 20 - 25°C for 2 hrs. After completion of the reaction, the reaction solvent was removed by vacuum concentration, and then trifluoroacetic acid was completely removed by adding 5 mL of toluene twice to obtain a concentrated TFA salt of the title compound. |
46% | Inert atmosphere | To stirred solution of 6-maleimidocaproyl-val-cit-PAB 38 in dry DMF under nitrogen, bis-(p-nitrophenyl)carbonate was added followed by DIPEA, resulting in a colour change from colourless to bright yellow. The solution was stirred at room temperature under nitrogen for 1 h after which the DMF was removed by high vacuum to give an oily residue. This was triturated with ethyl acetate for 15 min resulting in precipitation which was completed by the addition of ether. The solid was collected and washed well with ether and air dried to give an off-white solid. TLC [silica gel: 10 percent MeOH/DCM Rf0.46].This was purified by chromatography [silica gel: 5-10percent MeOH/DCM gradient elution] to give the activated linker 39 as a white solid 0.006 g, (46percent). MS (m/z) 738..3091 (M+H), HRMS (m/z) calculated for C35H43N7O11Na M+Na 760.2918 found 760.2922 |
40% | With pyridine In dichloromethane at 0 - 20℃; for 3.16667 h; Inert atmosphere | 168.6 mg (0.294 mmol) of mc-vc-PABOH was dissolved in 5 ml of anhydrous pyridine under nitrogen protection.Cool to about 0°C. Another 179 mg (3 eq) of PNP was dissolved in 5 ml of DCM, and it was slowly added to the reaction system. And inThe ice bath was removed after keeping at 0° C. for 10 min, and the reaction was stirred at room temperature for 3 h. After the reaction is completed, add 70ml EA and 100ml 15percentAqueous citric acid, dispense organic layer. The organic layer was washed successively with citric acid, water, saturated saline and dried over anhydrous sodium sulfate.Dry and filter. The filtrate was concentrated to dryness under reduced pressure to give a pale yellow oil. Methyl tert-butyl ether was added for crystallization to give an off-white solid.86mg, yield 40percent. |
40% | With pyridine In dichloromethane at 0 - 20℃; for 3.16667 h; Inert atmosphere | Under the protection of nitrogen, 168.6 mg (0.294 mmol) of mc-vc-PABOH was dissolved in 5 ml of anhydrous pyridine.The reaction system was cooled to about 0 °C. Further, PNP 179 mg (3 eq) was dissolved in 5 ml of DCM, and then slowly added to the reaction system.And after 10 minutes at 0 ° C, remove the ice bath.The reaction was further stirred at room temperature for 3 h. The reaction is completed,Add 70 ml EA and 100 ml 15percent aqueous citric acid solution.The organic layer was separated. The organic layer was washed successively with citric acid, water and saturated brine.Dry over anhydrous sodium sulfate and filter.The filtrate was concentrated to dryness under reduced pressure to give a pale yellow oil.Crystallization with methyl tert-butyl ether gave 86 mg of an off-white solid.The yield was 40percent. |
Yield | Reaction Conditions | Operation in experiment |
---|---|---|
100% | In 1-methyl-pyrrolidin-2-one; for 24h;Heating; | vc-PABOH 205 mg (0.54 mmol) and MC-OSu 184 mg (1.1 eq) were added to 10 ml of NMP.The reaction was stirred for 24 h under warming. After completion of the reaction, the mixture was concentrated under reduced pressure at 40 C., and 20 ml of methyl tert-butyl ether was added to the resulting oil.Crystallization. It was filtered and the cake was washed with methyl tert-butyl ether to give 310 mg of product in 100% yield. |
100% | In 1-methyl-pyrrolidin-2-one; at 20℃; for 24h; | VP-PABOH 205 mg (0.54 mmol) and MC-OSu 184 mg (1.1 eq) were added to 10 ml of NMP, and the reaction was stirred at room temperature for 24 h.The reaction is completed,Should be completed, concentrated at 40 C under reduced pressure,The obtained oil was added to 20 ml of methyl t-butyl ether and stirred to crystallize. filter,The filter cake was washed with methyl tert-butyl ether.Get 310mg of product,The yield is 100%. |
97% | In N,N-dimethyl-formamide; at 20 - 30℃; | To mc-OSu (1.7 eq.) was added anhydrous N,N-Dimethylformamide (3 vol.) and the resulting mixture was stirred at 20C until a clear colorless solution formed. A solution of Val-Cit-PABOH (1 eq.) in anhydrous N,N-Dimethylformamide (7 vol.) was then added over 30 minutes while keeping temperature below 30C. The reaction mixture was stirred at 30C for 5-6 hours until reaction was complete. Ethyl acetate (30 vol.) was then added over 30 minutes at 30C. The resulting suspension was stirred at that temperature for 10-20 minutes before being cooled to 20C and stirred at 20C for 2-4 hours. Filtered solids were solubilized in N.N-Dimethylformamide (10 vol.) and the resulting mixture was stirred at 30C for 30-60 minutes. Ethyl acetate (30 vol.) was added over 30 minutes at 30C. The resulting suspension was stirred at that temperature for 10-20 minutes before being cooled to 20C and stirred at 20C for 2-4 hours. The resulting solids were collected by filtration, washed and dried under vacuum (97% yield). MS: m/e 573 (MH)+, 595 (M+Na)+. |
96% | In N,N-dimethyl-formamide; for 2h; | [0438] Fmoc-val-cit-PAB-OH (14.61 g, 24.3 mmol, 1.0 eq.; see, e.g., U.S. Patent No. 6,214,345 to Firestone et al.) was diluted with DMF (120 mL, 0.2 M) and to this solution was added a diethylamine (60 mL). The reaction was monitored by HPLC and found to be complete in 2 h. The reaction mixture was concentrated and the resulting residue was precipitated using ethyl acetate (ca. 100 mL) under sonication over for 10 min. Ether (200 mL) was added and the precipitate was further sonicated for 5 min. The solution was allowed to stand for 30 min. without stirring and was then filtered and dried under high vacuum to provide Val-cit-P AB-OH, which was used in the next step without further purification. Yield: 8.84 g (96%). Val-cit-PAB-OH (8.0 g, 21 mmol) was diluted with DMF (110 mL) and the resulting solution was treated with MC-OSu (Willner et at, 1993, Bioconjugate Chem. 4:521; 6.5 g, 21 mmol, 1.0 eq.). The reaction was complete according to HPLC after 2 h. The reaction mixture was concentrated and the resulting oil was precipitated using ethyl acetate (50 mL). After sonicating for 15 min, ether (400 mL) was added and the mixture was sonicated further until all large particles were broken up. The solution was then filtered and the solid dried to provide an off-white solid intermediate. Yield: 11.63 g (96%); ES-MS m/z 757.9 [M-H] |
96% | In N,N-dimethyl-formamide; for 2h; | [0454] Fmoc-val-cit-P AB-OH (14.61 g, 24.3 mmol, 1.0 eq., see, e.g., U.S. Patent No. 6,214,345 to Firestone et al.) was diluted with DMF (120 mL, 0.2 M) and to this solution was added a diethylamine (60 mL). The reaction was monitored by HPLC and found to be complete in 2 h. The reaction mixture was concentrated and the resulting residue was precipitated using ethyl acetate (ca. 100 mL) under sonication for 10 min. Ether (200 mL) was added and the precipitate was further sonicated for 5 min. The solution was allowed to stand for 30 min. without stirring and was then filtered and dried under high vacuum to provide Val-cit-P AB-OH, which was used in the next step without further purification. Yield: 8.84 g (96%). Val-cit-P AB-OH (8.0 g, 21 mmol) was diluted with DMF (110 mL) and the resulting solution was treated with MC-OSu (Willner et al, 1993, EPO <DP n="138"/>Bioconjugate Chem. 4:521; 6.5 g, 21 mmol, 1.0 eq.). The reaction was complete according to HPLC after 2 h. The reaction mixture was concentrated and the resulting oil was precipitated using ethyl acetate (50 mL). After sonicating for 15 min, ether (400 niL) was added and the mixture was sonicated further until all large particles were broken up. The solution was then filtered and the solid dried to provide an off-white solid intermediate. Yield: 11.63 g (96%); ES-MS m/z 757.9 [M-H] |
96% | In N,N-dimethyl-formamide; for 2h; | Fmoc-val-cit-PAB-OH (14.61 g, 24.3 mmol), 1.0 eq., U.S. Patent No. 6214345 to Firestone et al.) was dilutedwith DMF (120 mL, 0.2 M) and to this solution was added a diethylamine (60 mL). The reaction was monitored by HPLCand found to be complete in 2 h. The reaction mixture was concentrated and the resulting residue was precipitated usingethyl acetate (ca. 100 mL) under sonication over for 10 min. Ether (200 mL) was added and the precipitate was furthersonicated for 5 min. The solution was allowed to stand for 30 min. without stirring and was then filtered and dried under high vacuum to provide Val-cit-PAB-OH, which was used in the next step without further purification. Yield: 8.84 g (96%).Val-cit-PAB-OH (8.0 g, 21 mmol) was diluted with DMF (110 mL) and the resulting solution was treated with MC-OSu(Willner et al., (1993) Bioconjugate Chem. 4:521; 6.5 g, 21 mmol, 1.0 eq.). Reaction was complete according to HPLCafter 2 h. The reaction mixture was concentrated and the resulting oil was precipitated using ethyl acetate (50 mL). Aftersonicating for 15 min, ether (400 mL) was added and the mixture was sonicated further until all large particles werebroken up. The solution was then filtered and the solid dried to provide an off-white solid intermediate. Yield: 11.63 g(96%); ES-MS m/z 757.9 [M-H] |
92% | In 1-methyl-pyrrolidin-2-one; at 20℃; for 16h;Inert atmosphere; | Mc-Val-Cit-PABOH 5. Compound 3 (0.19 g, 0.49 mmol) was added to a solution of ester 4 (0.18 g, 0.57 mmol) in NMP (7 mL), and the reaction was stirred for 16 h. The solvent was evaporated at reduced pressure, and then the residue was triturated with Et20 (30 mL). The crude product was collected by filtration and washed with Et20 (3 x 15 mL). The crude product was purified by flash chromatography using a pre-packed 25 g silica column [solvent A: MeOH; solvent B: CH2C12; gradient: 0%A / 100%B (1 CV), 0%A / 100%B? 30%A / 70%B (10 CV), 30%A / 70%B (2 CV); flow rate: 25 mL/min; monitored at 254 and 280 nm] to afford linker 5 (0.26 g, 0.45 mmol, 92% yield) as a brown solid. [000197] NMR (500 MHz, DMSO-d6) delta 9.82 (1H, s), 7.98 (1H, d,J= 8.5 Hz), 7.73 (1H, d,J= 8.5 Hz), 7.46 (2H, d,J= 7.8 Hz), 7.14 (2H, d, 7= 8.0 Hz), 6.92 (2H, s), 5.89 (1H, t,J= 6.0 Hz), 5.33 (2H, s), 5.01 (1H, t, J= 5.7 Hz), 4.34 (2H, d, J= 4.5 Hz), 4.29 (1H, q,J= 7.4 Hz), 4.14 - 4.05 (1H, m), 3.29 (2H, t, .7=6.3 Hz), 2.99-2.82 (2H, m), 2.17- 1.96 (1H, m), 1.94- 1.80 (2H, m), 1.67-1.03 (8H, m), 0.76 (3H, d, .7=6.4 Hz), 0.73 (3H, d, .7=6.1 Hz). [000198] 13C NMR (125 MHz, DMSO-d6) delta 173.2, 172.7, 171.5, 170.8, 159.3, 138.0, 137.8, 134.9, 127.3, 119.2, 63.0, 58.0, 53.5, 37.4, 35.4, 30.8, 29.8, 28.2, 26.2, 25.7, 25.4, 19.7, 18.6. |
0.69 g | In N,N-dimethyl-formamide; at 20℃; | The product of reaction step d (0.5 g, 1.3 mmol) was dissolved in 18 mL of DMF and added to MC-OSu (0.45 g,1.4 mmol), the reaction was stirred at room temperature overnight. After the reaction is completed, the reaction solvent is spin-dried, washed with diethyl ether three times, and filtered to obtain a yellow solid.The body was 0.69 g without further purification. |
With triethylamine; In N,N-dimethyl-formamide; at 20℃; for 20h; | : Procedure A: A solution of compound 8 (0.270 g, 0.450 mmol, 1.0 equiv) in DMF (0.2 M) was treated with triethylamine (1.25 mL, 8.90 mmol, 19.8 equiv) followed by stirring at room temperature for 24 hours. DMF and excess triethylamine were removed under reduced pressure. The resulting residue was dissolved in DMF (0.1 M) and to the solution Mc-OSu (0.150 g, 0.500 mmol, 1.1 equiv) was added. The reaction mixture was stirred at room temperature for 20 hours. DMF was removed under reduced pressure and the resulting residue was purified by flash column chromatography using 3-12% MeOH-CH2Cl2 as solvent. The product was obtained as white solid. Yield: 86%. |
Yield | Reaction Conditions | Operation in experiment |
---|---|---|
95% | With N-ethyl-N,N-diisopropylamine; In N,N-dimethyl-formamide; at 25℃; | mc-Val-Cit-PABOH (1 eq.) was mixed with bis(4-nitrophenyl) carbonate (1.87 eq.) in N,N-dimethylformamide (8 vol.) at 20C. N,N-diisopropyl ethyl amine (1.75 eq.) was added at 25C. The reaction mixture was stirred at 25C for 2-6 hours until reaction was complete. Product was precipitated out of the reaction mixture by adding anhydrous ethyl acetate (12.5 vol.) at 25C and tert-Butyl methyl ether (12.5 vol.). The resulting slurry was stirred, then cooled to 0C and stirred for 10-30 minutes. The solids were isolated by filtration, washed and dried under vacuum before being re-slurried in ethyl acetate (12.5 vol.) at 20C, filtered and dried once more (95% yield). MS: m/e 738 (MH)+, 760 (M+Na)+. |
57% | With N-ethyl-N,N-diisopropylamine; In dichloromethane; N,N-dimethyl-formamide; at 23℃; for 20h; | To a solution of Compound 11 (500 mg, 0.87 mmol) and bis(4-nitrophenyl) carbonate (bis-PNP) (2.64 g, 8.72 mmol) in DCM:DMF (8:2, 25 mL) was added DIPEA (0.45 mL, 2.61 mmol). The reaction mixture was stirred for 20 h at 23 C and poured onto a silica gel column (DCM:CH3OH, from 50:1 to 10:1 ) to afford pure target Compound 9 (364 mg, 57%). Rf= 0.40 (CH2CI2:CH3OH, 9:1 ).1H NMR (400 MHz, CDCI3/CD3OD): delta 9.45 (s, 1 H), 8.23 (d, J = 8.3 Hz, 2H), 7.59 (d, J = 8.5 Hz, 2H), 7.35 (d, J = 8.3 Hz, 2H), 7.34 (d, J = 8.5 Hz, 2H), 6.65 (s, 2H), 5.20 (s, 2H), 4.56 (dt, J = 10.5, 5.4 Hz, 1 H), 4.15 (d, J = 7.2 Hz, 1 H), 3.46 (dd, J = 8.0, 6.4 Hz, 2H), 3.16-2.89 (m, 2H), 2.21 (dd, J = 8.3, 6.6 Hz, 2H), 2.06-1 .97 (m, 1 H), 1.90-1.83 (m, 1 H), 1.73-1.46 (m, 7H), 1.34- 1.20 (m, 2H), 0.91 (d, J = 6.7 Hz, 3H), 0.90 (d, = 6.7 Hz, 3H).13C NMR (125 MHz, CDCI3/CD3OD) delta 174.4, 172.4, 171.1 , 170.6, 160.5, 155.5, 152.5, 145.3, 138.7, 134.1 , 129.9, 129.5, 125.2, 121.8, 120.0, 70.6, 59.0, 53.2, 37.5, 35.8, 30.6, 29.6, 29.3, 28.1 , 26.2, 26.2, 25.1 , 19.1 , 18.1. ESI-MS m/z: Calcd. for CasH^N O^: 737.3. Found: 738.3 (M+H)+. |
57% | With N-ethyl-N,N-diisopropylamine; In N,N-dimethyl-formamide; at 0 - 20℃; for 15h;Inert atmosphere; | Preparation Example 1-2: Preparation of Compound (II-3) (0054) (0055) Under an argon stream, <strong>[159857-80-4]6-(2,5-dioxo-2,5-dihydro-1H-pyrrol-1-yl)-N-((S)-1-(((S)-1-((4-(hydroxymethyl)phenyl)amino)-1-oxo-5-ureidopentan-2-yl)amino)-3-methyl-1-oxobutan-2-yl)hexanamide</strong>(5.0 g, 8.7 mmol, Dubowchik et al., Bioconjugate Chem., 2002, 13 (4), pp 855-869) was dissolved in 60 mL of anhydrous dimethylformamide, added with N, N-diisopropylethylamine (3.0 mL, 17.4 mmol), and then cooled to 0C. To the mixture was added at once bis(4-nitrophenyl) carbonate (7.94 g, 26.1 mmol), followed by stirring at room temperature for 15 hrs. After completion of the reaction, the reaction mixture was concentrated in a high vacuum, and the concentrate was purified by silica gel column chromatography to obtain 4-((S)-2-((S)-2-(6-(2 ,5-dioxo-2,5-dihydro-1H-pyrrol-1-yl)hexanamido)-3-methylbutanamido)-5-ureidopentanamido)benzyl (4-nitrophenyl) carbonate (3.65 g, 57 %) as a pale yellow solid. (0056) 1H NMR (400 MHz, DMSO-d6) delta 0.83 (d, J = 6.8 Hz, 3 H), 0.86 (d, J = 6.8 Hz, 3 H), 1.09 (t, J = 7.2 Hz, 1 H), 1.19 (m, 2 H), 1.34 - 1.76 (m, 7 H), 1.96 (m, 1 H), 2.15 (m, 2 H), 2.99 (m, 2 H), 3.37 (m, 2 H), 4.19 (t, J= 7.8 Hz, 1 H), 4.39 (m, 1 H), 5.24 (s, 2 H), 5.41 (s, 2 H), 5.97 (brt, J= 5.6 Hz, 1 H), 7.00 (s, 2H), 7.41 (d, J= 8.4 Hz, 2 H), 7.57 (d, J= 7.2 Hz, 2 H), 7.65 (d, J= 8.4 Hz, 2 H), 7.80 (d, J= 8.4 Hz, 1 H), 8.09 (d, J = 7.2 Hz, 1 H), 8.31 (d, J = 7.2 Hz, 2 H), 10.05 (brs, 1 H) (0057) Under an argon stream, 4-((S)-2-((S)-2-(6-(2, 5-dioxo-2,5-dihydro-1H-pyrrol-1-yl)hexanamido)-3-methylbutanamido)-5-ureidopentanamido)benzyl (4-nitrophenyl) carbonate (3.65 g, 4.95 mmol) was dissolved in 90 mL of anhydrous dimethylformamide, and stirred, together with t-butyl (2-aminoethyl)(methyl)carbamate (0.86 g, 4.95 mmol), at 20 - 25C for 2 hrs. After completion of the reaction, the reaction mixture was completely concentrated in a high vaccum, and the concentrate was purified by silica gel column chromatography to obtain an amino-protected derivative of compound (II-3) (3.8 g, 99%). (0058) LC-MS m/z: 773.5 [M+H]+ (0059) To a solution of the amino-protected derivative of compound (II-3) (146 mg, 0.186 mmol) in 5 mL of dichloromethane was dropwise added 2 mL of trifluoroacetic acid, followed by stirring at 20 - 25C for 2 hrs. After completion of the reaction, the reaction solvent was removed by vacuum concentration, and then trifluoroacetic acid was completely removed by adding 5 mL of toluene twice to obtain a concentrated TFA salt of the title compound. |
57% | With N-ethyl-N,N-diisopropylamine; In dichloromethane; N,N-dimethyl-formamide; at 23℃; for 20h; | To a solution of LIN 1 -2 (500 mg, 0.87 mmol) and bis(4-nitrophenyl) carbonate (bis-PNP) (2.64 g, 8.72 mmol) in DCM:DMF (8:2, 25 mL) was added DIPEA (0.45 mL, 2.61 mmol). The reaction mixture was stirred for 20 h at 23 SC and poured onto a silica gel column (DCM:CH30H, from 50:1 to 10:1 ) to afford pure target LIN 1 (364 mg, 57%). (1575) Rf= 0.40 (CH2Cl2:CH3OH, 9:1 ). (1576) 1 H NMR (400 MHz, CDCI3/CD3OD): d 9.45 (s, 1 H), 8.23 (d, J = 8.3 Hz, 2H), 7.59 (d, J = 8.5 Hz, 2H), 7.35 (d, J = 8.3 Hz, 2H), 7.34 (d, J = 8.5 Hz, 2H), 6.65 (s, 2H), 5.20 (s, 2H), 4.56 (dt, J = 10.5, 5.4 Hz, 1 H), 4.15 (d, J = 7.2 Hz, 1 H), 3.46 (dd, J = 8.0, 6.4 Hz, 2H), 3.16-2.89 (m, 2H), 2.21 (dd, J = 8.3, 6.6 Hz, 2H), 2.06-1 .97 (m, 1 H), 1 .90-1 .83 (m, 1 H), 1 .73-1 .46 (m, 7H), 1 .34-1 .20 (m, 2H), 0.91 (d, J = 6.7 Hz, 3H), 0.90 (d, J = 6.7 Hz, 3H). (1577) 13C NMR (125 MHz, CDCI3/CD3OD) d 174.4, 172.4, 171 .1 , 170.6, 160.5, 155.5, 152.5, 145.3, 138.7, 134.1 , 129.9, 129.5, 125.2, 121 .8, 120.0, 70.6, 59.0, 53.2, 37.5, 35.8, 30.6, 29.6, 29.3, (1578) 28.1 , 26.2, 26.2, 25.1 , 19.1 , 18.1 . (1579) ESI-MS m/z : Calcd. for C35H43N7O1 1 : 737.3. Found: 738.3 (M+H)+. |
46% | In N,N-dimethyl-formamide;Inert atmosphere; | To stirred solution of 6-maleimidocaproyl-val-cit-PAB 38 in dry DMF under nitrogen, bis-(p-nitrophenyl)carbonate was added followed by DIPEA, resulting in a colour change from colourless to bright yellow. The solution was stirred at room temperature under nitrogen for 1 h after which the DMF was removed by high vacuum to give an oily residue. This was triturated with ethyl acetate for 15 min resulting in precipitation which was completed by the addition of ether. The solid was collected and washed well with ether and air dried to give an off-white solid. TLC [silica gel: 10 % MeOH/DCM Rf0.46].This was purified by chromatography [silica gel: 5-10% MeOH/DCM gradient elution] to give the activated linker 39 as a white solid 0.006 g, (46%). MS (m/z) 738..3091 (M+H), HRMS (m/z) calculated for C35H43N7O11Na M+Na 760.2918 found 760.2922 |
40% | With pyridine; In dichloromethane; at 0 - 20℃; for 3.16667h;Inert atmosphere; | 168.6 mg (0.294 mmol) of mc-vc-PABOH was dissolved in 5 ml of anhydrous pyridine under nitrogen protection.Cool to about 0C. Another 179 mg (3 eq) of PNP was dissolved in 5 ml of DCM, and it was slowly added to the reaction system. And inThe ice bath was removed after keeping at 0 C. for 10 min, and the reaction was stirred at room temperature for 3 h. After the reaction is completed, add 70ml EA and 100ml 15%Aqueous citric acid, dispense organic layer. The organic layer was washed successively with citric acid, water, saturated saline and dried over anhydrous sodium sulfate.Dry and filter. The filtrate was concentrated to dryness under reduced pressure to give a pale yellow oil. Methyl tert-butyl ether was added for crystallization to give an off-white solid.86mg, yield 40%. |
40% | With pyridine; In dichloromethane; at 0 - 20℃; for 3.16667h;Inert atmosphere; | Under the protection of nitrogen, 168.6 mg (0.294 mmol) of mc-vc-PABOH was dissolved in 5 ml of anhydrous pyridine.The reaction system was cooled to about 0 C. Further, PNP 179 mg (3 eq) was dissolved in 5 ml of DCM, and then slowly added to the reaction system.And after 10 minutes at 0 C, remove the ice bath.The reaction was further stirred at room temperature for 3 h. The reaction is completed,Add 70 ml EA and 100 ml 15% aqueous citric acid solution.The organic layer was separated. The organic layer was washed successively with citric acid, water and saturated brine.Dry over anhydrous sodium sulfate and filter.The filtrate was concentrated to dryness under reduced pressure to give a pale yellow oil.Crystallization with methyl tert-butyl ether gave 86 mg of an off-white solid.The yield was 40%. |
With N-ethyl-N,N-diisopropylamine; In N,N-dimethyl-formamide; for 1h; | [0439] The off-white solid intermediate (8.0 g, 14.0 mmol) was diluted with DMF (120 mL, 0.12 M) and to the resulting solution was added bis(4-nitrophenyl)carbonate (8.5 g, 28.0 mmol, 2.0 eq.) and D3EA (3.66 mL, 21.0 mmol, 1.5 eq.). The reaction was complete in 1 h according to HPLC. The reaction mixture was concentrated to provide an oil that is precipitated with EtOAc, and then triturated with EtOAc (ca. 25 mL). The solute was further precipitated with ether (ca. 200 mL) and triturated for 15 min. The solid was filtered and dried under high vacuum to provide Compound AB which is 93% pure EPO <DP n="168"/>according to HPLC and used in the next step without further purification. Yield: 9.7 g (94%). | |
With N-ethyl-N,N-diisopropylamine; In N,N-dimethyl-formamide; for 1h; | [0455] The off-white solid intermediate (8.0 g, 14.0 mmol) was diluted with DMF (120 mL, 0.12 M) and to the resulting solution was added bis(4-nitrophenyl)carbonate (8.5 g, 28.0 mmol, 2.0 eq.) and DIEA (3.66 mL, 21.0 mmol, 1.5 eq.). The reaction was complete in 1 h according to HPLC. The reaction mixture was concentrated to provide an oil that was precipitated with EtOAc, and then triturated with EtOAc (ca. 25 mL). The solute was further precipitated with ether (ca. 200 mL) and triturated for 15 min. The solid was filtered and dried under high vacuum to provide Compound AB which was 93% pure according to HPLC and used in the next step without further purification. Yield: 9.7 g (94%). | |
With N-ethyl-N,N-diisopropylamine; In N,N-dimethyl-formamide; for 1h; | The off-white solid intermediate (8.0 g, 14.0 mmol) was diluted with DMF (120 mL, 0.12 M) and to the resultingsolution was added bis(4-nitrophenyl)carbonate (8.5 g, 28.0 mmol, 2.0 eq.) and DIEA (3.66 mL, 21.0 mmol, 1.5 eq.).The reaction was complete in 1 h according to HPLC. The reaction mixture was concentrated to provide an oil that wasprecipitated with EtOAc, and then triturated with EtOAc (ca. 25 mL). The solute was further precipitated with ether (ca.200 mL) and triturated for 15 min. The solid was filtered and dried under high vacuum to provide Compound AB whichwas 93% pure according to HPLC and used in the next step without further purification. Yield: 9.7 g (94%). | |
0.42 g | With N-ethyl-N,N-diisopropylamine; In N,N-dimethyl-formamide; at 20℃; | dissolving the reaction product of reaction step e in 12 mL of DMF,After dissolution, (PNP) 2CO (1.1 g, 3.6 mmol) and DIEA 0.5 mL were added.Stir at room temperature overnight. After the reaction is completed, the solvent is evaporated.Separation and purification on a silica gel column, the separation conditions are dichloromethane:Methanol = 20:1.The product is a yellowish solid0.42g. |
With N-ethyl-N,N-diisopropylamine; In N,N-dimethyl-formamide; at 20℃; | Compound 7 (1.3 g, 2.16 rnrnoi) and bis(4~nitrophenyl) carbonate (1.34 g, 4.34 rnrnol) were combined and dissolved in DMF (6 mb). To this solution was added DIPEA (0.75 mL, 4.35 mmol). The mixture was stirred overnight at room temperature. The reaction mixture was evaporated to dryness and the residue was washed with ether. |
Yield | Reaction Conditions | Operation in experiment |
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67% | With benzotriazol-1-ol; diisopropyl-carbodiimide; In dichloromethane; N,N-dimethyl-formamide; at 23℃; for 2h; | To a solution of Compound 10 (1 .6 g, 3.42 mmol) and 4-aminobenzyl alcohol (PABOH) (0.84 g, 6.84 mmol) in DCM (60 mL) was added a solution of HOBt (0.92 g, 6.84 mmol) in DMF (5 mL). DIPCDI (1.05 mL, 6.84 mmol) was added, the reaction mixture was stirred for 2 h at 23 C, Et20 (150 mL) was added, and the solid obtained was filtrated in a filter plate under vacuum to obtain Compound 11 (1 .31 g, 67%).1H NMR (500 MHz, DMSO-d6): delta 9.88 (s, 1 H), 8.03 (d, J = 7.6 Hz, 1 H), 7.77 (dd, J = 12.2, 8.5 Hz, 1 H), 7.53 (d, J = 8.2 Hz, 2H), 7.21 (d, J = 8.2 Hz, 2H), 6.99 (s, 3H), 6.01 -5.92 (m, 1 H), 5.39 (s, 2H), 5.07 (s, 1 H), 4.41 (s, 2H), 4.39-4.31 (m, 1 H), 4.23- 4.12 (m, 1 H), 3.36 (t, J = 7.0 Hz, 2H), 3.06-2.97 (m, 1 H), 2.96-2.90 (m, 1 H), 2.22-2.03 (m, 2H), 2.01 -1 .88 (m, 1 H), 1 .76-1.62 (m, 1 H), 1 .63-1 .28 (m, 6H), 1 .25-1.1 1 (m, 2H), 0.84 (d, J = 6.9 Hz, 3H), 0.81 (d, J = 6.8 Hz, 3H).ESI-MS m/z: Calcd. for C28H4oN607: 572.3. Found: 573.3 (M+H)+. |
67% | With benzotriazol-1-ol; diisopropyl-carbodiimide; In dichloromethane; N,N-dimethyl-formamide; at 23℃; for 2h; | To a solution of LIN 1-1 (1.6 g, 3.42 mmol) and 4-aminobenzyl alcohol (PABOH) (0.84 g, 6.84 mmol) in DCM (60 mL) was added a solution of HOBt (0.92 g, 6.84 mmol) in DMF (5 mL). DIPCDI (1.05 mL, 6.84 mmol) was added, the reaction mixture was stirred for 2 h at 23 SC, Et20 (150 mL) was added, and the solid obtained was filtrated in a filter plate under vacuum to obtain LIN 1-2 (1.31 g, 67%). (1570) 1H NMR (500 MHz, DMSO-ofe): d 9.88 (s, 1H), 8.03 (d, J= 7.6 Hz, 1H), 7.77 (dd, J= 12.2, 8.5 Hz, 1 H), 7.53 (d, J = 8.2 Hz, 2H), 7.21 (d, J= 8.2 Hz, 2H), 6.99 (s, 3H), 6.01-5.92 (m, 1H), 5.39 (s, 2H), 5.07 (s, 1H), 4.41 (s, 2H), 4.39-4.31 (m, 1H), 4.23-4.12 (m, 1H), 3.36 (t, J= 7.0 Hz, 2H), 3.06-2.97 (m, 1H), 2.96-2.90 (m, 1H), 2.22-2.03 (m, 2H), 2.01-1.88 (m, 1H), 1.76- 1.62 (m, 1 H), 1.63-1.28 (m, 6H), 1.25-1.11 (m, 2H), 0.84 (d, J= 6.9 Hz, 3H), 0.81 (d, J= 6.8 Hz, 3H). (1571) ESI-MS m/z : Calcd. for C28H40N6O7: 572.3. Found: 573.3 (M+H)+. |
Yield | Reaction Conditions | Operation in experiment |
---|---|---|
Compound 20-8 (0.768 g, 0.883 mmol) was added to the reaction mixture of Compound 23-3 as just described above. The reaction mixture was reacted for about 5min and then was purified by flash chromatography using methanol-dichloromethane gradient to give Compound 23-4, the 20 site of which was protected by Boc. Then the protecting group Boc was de-protected by treatment with TFA and the product was recovered by precipitation with diethyl ether to give the TFA salt of Compound 23-4. |
Yield | Reaction Conditions | Operation in experiment |
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80% | With thionyl chloride; In 1-methyl-pyrrolidin-2-one; at 0 - 25℃; for 4h;Inert atmosphere; | Dry N-methylpyrrolidone, NMP (55 g) and 6-(2,5-dioxo-2,5-dihydro-lH-pyrrol-l-yl)- N-((S)-l-(((S)-l-((4-(hydroxymethyl)phenyl)amino)-l-oxo-5-ureidopentan-2-yl)amino)-3- methyl-l-oxobutan-2-yl)hexanamide, <strong>[159857-80-4]MC-VC-PAB</strong>-OH (10.0 g, 17.5 mmol) were charged to the reactor under N2. (0143) (0144) The contents were warmed to 50-55 C for 1 h before cooling to 0-5 C. Thionyl chloride (2.6 g, 21.9 mmol) was charged to the reactor over 1 h with temperature of <5 C. After the addition, the reactor contents were warmed to 20-25 C and held for 1 h. Since LC showed starting material remaining, the batch was cooled to 0-5 C and more thionyl chloride (0.2 g, 1.7 mmol) was charged to the reactor over 1 h. After the addition, the contents were warmed to 20-25 C and held for 1 h. The batch was cooled to 0-5 C and water (170 g) was added while maintaining temperature of <5 C. The resultant slurry was filtered and the filter cake washed with water (2 x 30 g). The cake was then washed with EtOAc (30 g), CH3CN (2 x 30 g) and MTBE (1 x 30 g). The wet cake was dried under vacuum at 20-25 C to give a lightly colored solid (8.3 g, 80% yield). lH NMR (600 MHz, DMSO-d6, 28 C) delta 10.02 (s, IH), 8.06 (d, 7.5, IH), 7.78 (d 8.7, IH), 7.60 (d, 8.6, 2H), 7.36 (d, 8.6, 2H), 6.99 (s, 2H), 5.98 (bs, IH), 5.40 (vbs, IH), 4.71 (s, 2H), 4.38 (m, IH), 4.19 (dd, .5, 6.9, IH), 3.37 (t, 7.1, 2H), 3.03 (m, IH), 2.94 (m, IH), 2.18 (m, IH), 2.12 (m, IH), 1.97 (m, IH), 1.70 (m, IH), 1.60 (m, IH), 1.48 (m, 5H), 1.37 (m, IH), 1.19 (pen, 7.7, 2H), 0.85 (d, 6.8, 3H), 0.82 (d, 6.8, 3H). 13C NMR (151 MHz, DMSO-d6, 28 deg C) delta 172.2, 171.2, 170.9, 170.6, 158.8, 138.9, 134.3, 132.2, 129.4, 119.0, 57.5, 53.0, 46.1, 38.5, 36.9, 34.8, 30.3, 29.2, 27.7, 26.7, 25.7, 24.8, 19.1, 18.1. |
71% | With thionyl chloride; at 20℃; for 1.5h; | To a stirred suspension of R18 (100 mg, 0.174 mmol, 1.0 eq) at room temperature was slowly added SOCl2(14 muL, 0.192 mmol, 1.1 eq) using a micro syringe. The slurry reaction mixture was stirred for 1.5 h and an aliquot analyzed by LC/MS indicated the formation of the desired. The crude mixture was concentrated to remove all volatiles under reduced pressure. The mixture was diluted with 2 mL of DMSO and loaded on to an ISCO C18 Aq 50g column for purification (10 - 95% MeCN in water, 0.05% AcOH). The pure fractions were combined and lyophilized to give 72 mg (71%) of R19 as an off-white solid. for C28H39ClN6O6, 590.26; found 591.3 (M+H), 1181.5 (2M+H).1H NMR (500 MHz; DMSO-d6) ' 10.03 (s, 1H), 8.03 - 8.11 (m, 1H), 7.79 (d, J = 8.30 Hz, 1H), 7.57 - 7.63 (m, J = 8.79 Hz, 2H), 7.34 - 7.38 (m, J = 8.79 Hz, 2H), 6.99 - 7.02 (m, 2H), 5.97 (br. s., 1H), 5.40 (br. s., 2H), 4.71 (s, 2H), 4.34 - 4.43 (m, 2H), 4.16 - 4.21 (m, 1H), 3.36 - 3.42 (m, 3H), 2.90 - 3.06 (m, 3H), 2.07 - 2.22 (m, 3H), 1.91 - 2.00 (m, 1H), 1.66 - 1.73 (m, 1H), 1.31 - 1.41 (m, 1H), 1.18 (quin, J = 7.69 Hz, 3H), 0.79 - 0.89 (m, 7H). |
With thionyl chloride; N,N-dimethyl-formamide; In dichloromethane; at 20℃;Sealed tube; | 6-(2,5-Dioxo-2,5-dihydro-1H-pyrrol-1-yl)-N-((S)-1-((S)-1-(4-(hydroxymethyl)phenylamino)-1-oxo-5-ureidopentan-2-ylamino)-3-methyl-1-oxobutan-2-yl)hexanamide 8, prepared according to procedures in WO 2012113847; U.S. Pat. No. 7,659,241; U.S. Pat. No. 7,498,298; US 20090111756; US 20090018086; U.S. Pat. No. 6,214,345; Dubowchik et al (2002) Bioconjugate Chem. 13(4):855-869 (1.009 g, 1.762 mmol, 1.000, 1009 mg) was taken up in N,N-dimethylformamide (6 mL, 77 mmol, 44, 5700 mg). To this was added a solution of thionyl chloride (1.1 equiv., 1.938 mmol, 1.100, 231 mg) in dichloromethane (DCM) (1 mL, 15.44 mmol, 8.765, 1325 mg) in portions dropwise ( was added over 1 hour, stirred one hour at room temperature (RT) then added the other half over another hour). The solution remained a yellow color. Another 0.6 eq of thionyl chloride was added as a solution in 0.5 mL DCM dropwise, carefully. The reaction remained yellow and was stirred sealed overnight at RT. The reaction was monitored by LC/MS to 88% product benzyl chloride 9. Another 0.22 eq of thionyl chloride was added dropwise as a solution in 0.3 mL DCM. When the reaction approached 92% benzyl chloride 9, the reaction was bubbled with N2. The concentration was reduced from 0.3 M to 0.6 M. The product N-((S)-1-((S)-1-(4-(chloromethyl)phenylamino)-1-oxo-5-ureidopentan-2-ylamino)-3-methyl-1-oxobutan-2-yl)-6-(2,5-dioxo-2,5-dihydro-1H-pyrrol-1-yl)hexanamide 9 was stored in the refrigerator as a 0.6 M solution and used as is. M/Z 591.3, 92% yield. |
With thionyl chloride; In N,N-dimethyl-formamide; | <strong>[159857-80-4]MC-Val-Cit-PAB</strong>-Cl was obtained from the chlorination of <strong>[159857-80-4]MC-Val-Cit-PAB</strong>-OH. <strong>[159857-80-4]MC-Val-Cit-PAB</strong>-OHSee patent "Antibody drug con jugates, W02014/191578A1The obtained <strong>[159857-80-4]MC-Val-Cit-PAB</strong>-OH was dissolved in anhydrous DMF, and thionyl chloride (2 eq.The DMF was removed under reduced pressure, and the reaction product was separated on silica gel column to afford <strong>[159857-80-4]MC-Val-Cit-PAB</strong>-Cl |
Yield | Reaction Conditions | Operation in experiment |
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98% | In 1-methyl-pyrrolidin-2-one; at 20℃; for 16h;Inert atmosphere; | To a stirred solution of val-cit-PAB 24 (0.11 g, 0.29 mmol) in dry N- methylpyrrolidinone, NMP (5 ml) under nitrogen, N-succinimidyl-6- maleimidohexanoate (0.0983 g, 0.318 mmol) was added and the resulting light- brown solution stirred at room temperature for 16 h. The NMP was removed by high vacuum at < 40C. The resulting thick oily residue was triturated with dry ether (20 ml) the solid collected by filtration and washed several times with dry ether and air dried to give the desired product 38 an off-white powder 0.16 g (98%). TLC [silica gel: 10% MeOH/DCM Rf0.21. MS (m/z) 572.653 (M+1) calculated for |
Yield | Reaction Conditions | Operation in experiment |
---|---|---|
1 g | With triethylamine; In dichloromethane; N,N-dimethyl-formamide; at 0℃; | To a solution of triphosgene (156 mg, 0.52 mmol) in DCM (20 mL) was added a solution of 1 (1.0 g, 1.05 mmol) and Et3N (318 mg, 3.15 mmol) in DCM (5.0 mL). The mixture was stirred at 0 C. for 1 h, and concentrated to give the crude intermediate, which was added (0.88 g, 1.53 mmol) in DCM (20 mL) was added to a mixture of triethylamine (310 mg, 3.07 mmol) and <strong>[159857-80-4]6-(2,5-dioxo-2,5-dihydro-1H-pyrrol-1-yl)-N-((S)-1-(((S)-1-((4-(hydroxymethyl)phenyl)amino)-1-oxo-5-ureidopentan-2-yl)amino)-3-methyl-1-oxobutan-2-yl)hexanamide</strong>, <strong>[159857-80-4]MC-VC-PAB</strong> (1.0 g, 1.02 mmol) in DMF (10 mL) at 0 C. The mixture was diluted with DCM (40 mL), washed with water (2×30 mL). The organic layer was dried over Na2SO4, concentrated and purified by chromatography on silica (0-10% MeOH in DCM) to give tert-butyl (2-((S)-2-(((tert-butyldimethylsilyl)oxy)methyl)-4-methylenepyrrolidine-1-carbonyl)-5-((5-(4-((S)-2-(((tert-butyldimethylsilyl)oxy)methyl)-4-methylenepyrrolidine-1-carbonyl)-5-((((4-((S)-2-((S)-2-(6-(2,5-dioxo-2,5-dihydro-1H-pyrrol-1-yl)hexanamido)-3-methylbutanamido)-5-ureidopentanamido)benzyl)oxy)carbonyl)amino)-2-methoxyphenoxy)pentyl)oxy)-4-methoxyphenyl)carbamate 21 (1.0 g, 0.64 mmol, 62.4% yield) as a yellow solid. |
Yield | Reaction Conditions | Operation in experiment |
---|---|---|
0.013 g | In tetrahydrofuran; at 45℃; for 16h;Inert atmosphere; | Mc-Val-Cit-PABC- GPl 56 Carbamate 7. To a flask containing a solution of KGP156 (0.100 g, 0.281 mmol) in CH2C12 (5 mL) was added triphosgene (0.0417 g, 0.141 mmol), and the reaction was stirred for 18 h at ambient temperature. The solvent was evaporated by blowing with N2 gas followed by rotavapor evaporation at 40 C to obtain the isocyanate intermediate 10 as a tan solid. To the flask containing isocyanate 10 was added a solution of linker 5 (0.240 g, 0.420 mmol) in THF (2 mL), and the reaction was stirred for 16 h at 45 C. The solvent was evaporated under reduced pressure, and the resulting residue was purified by flash chromatography using a pre-packed 10 g silica column [solvent A: MeOH; solvent B: CH2C12; gradient: 0%A / 100%B ( 1 CV), 0%A / 100%B? 20%A / 80%B ( 10 CV), 20%A / 80%B (2 CV); flow rate: 12 mL/min; monitored at 254 and 280 nm] to afford a crude product. The crude product was further purified by preparative TLC (MeOH/CH2Cl2, 1 :9) followed by re- crystallization with MeOH to obtain desired carbamate 7 (0.013 g, 0.0136 mmol, 5% yield) as a tan solid. [000206] NMR (600 MHz, CD3OD) delta 7.63 (2H, d, J = 7.2 Hz), 7.42 (2H, d, J = 7.2 Hz), 6.93 (1 H, d, J = 8.5 Hz), 6.90 (1H, d, J = 8.8 Hz), 6.80 (2H, s), 6.55 (2H, s), 6.39 (1H, t, J= 7.3 Hz), 5.51 (2H, s), 5.17 (2H, s), 4.53 ( 1 H, t, J = 7.0 Hz), 4.18 (1H, d, 7 = 7.5 Hz), 3.84 (3H, s), 3.78 (3H, s), 3.77 (6H, s), 3.49 (2H, t, J = 7.1 Hz), 3.26 - 3.18 ( 1 H, m), 3.13 (1 H, dt, J = 13.3, 6.5 Hz), 2.66 (2H, t, J = 6.7 Hz), 2.29 (2H, t, J = 7.4 Hz), 2.17 - 2.10 (1 H, m), 2.09 (1 H, dt, 7 = 13.7, 6.8 Hz), 1.93 (2H, q, J = 12.3, 9.6 Hz), 1 .78 (1 H, ddt, ./ = 17.7, 13.3, 6.6 Hz), 1 .65 (3H, p, J = 7.4 Hz), 1 .59 (3H, p, J = 7.3 Hz), 1 .34 - 1 .29 (2H, m), 0.99 (6H, t, .7 = 7.4 Hz). [000207] 13C NMR ( 150 MHz, CD3OD) delta 176.3, 174.0, 172.6, 172.3, 162.3, 158.4, 156.2, 154.2, 144.2, 142.6, 140.0, 139.4, 138.4, 135.3, 135.3, 134.5, 134.2, 130.2, 129.6, 128.2, 121.1 , 109.8, 106.5, 67.4, 61 .2, 60.6, 56.5, 56.2, 55.0, 54.8, 38.4, 36.5, 35.0, 31.7, 30.4, 27.8, 27.4, 26.5, 26.4, 20.9, 19.8, 18.9. [000208] HRMS: Obsvd 976.4421 [M + Na]+, Calcd for C50H63N7NaO12: 976.4427. |
Yield | Reaction Conditions | Operation in experiment |
---|---|---|
28% | In N,N-dimethyl-formamide; at 20℃; for 72h;Inert atmosphere; | Synthesis of Mc-Val-Cit-PABC- GP05 : KGP05 isocyanate (.04 mmol) and Mc-Val-Cit- PAB-OH (.04 mmol) were added to an oven dried flask and dissolved in DMF (.8 ml). The reaction was allowed to stir under nitrogen at room temperature for 3 days. The solvent was removed by vacuum distillation and the resulting oil was purified by flash column chromatography (0-20% CH30H/CH2C12). The desired carbamate (.01 mmol, 28% yield) was obtained was a brown solid. |
With dibutyltin dilaurate; In N,N-dimethyl-formamide; at 20℃; | : A solution of KGP05 (0.232 g, 0.680 mmol, 1.0 equiv) in CH2Cl2 (0.1 M)- NaHCO3 Sat [1:1] was treated with triphosgene (0.403 g, 1.358 mmol, 2.0 equiv) followed by stirring at room temperature for 5 hours to generate the desired isocyanate. The reaction mixture was extracted with CH2Cl2 (3X). The combined organic layers were dried over sodium sulfate. The solvent was removed under reduced pressure, and the crude product was taken directly to the next step without further purification. Compound 1 (0.043 g, 0.075 mmol, 1.0 equiv) and the isocyanate compound 21 (0.040 g, 0.110 mmol, 1.5 equiv) were dissolved in DMF followed by addition of dibutyltin dilaurate (0.08 mL, 0.15 mmol, 2.0 equiv). The reaction was stirred at room temperature for 3 days. The solvent (DMF) was removed under reduced pressure and the resulting residue was purified by flash column chromatography using 2-15% MeOH-CH2Cl2 as solvent. The product was obtained as a white solid. Yield: 60%. 1H NMR (600 MHz, DMSO-d6) delta 10.01 (s, 1H), 8.68 (s, 1H), 8.10 (d, J = 7.2 Hz, 1H), 7.82 (d, J = 8.7 Hz, 1H), 7.62 (d, J = 8.7 Hz, 2H), 7.36 (d, J = 8.4 Hz, 2H), 7.00 (s, 2H), 6.88 (d, J = 8.7 Hz, 1H), 6.82 (d, J = 8.3 Hz, 1H), 6.55 (s, 2H), 6.03 - 5.93 (m, 2H), 5.42 (s, 2H), 5.04 (s, 2H), 4.38 (d, J = 7.8 Hz, 1H), 4.23 - 4.15 (m, 1H), 3.75 (s, 6H), 3.73 (s, 3H), 3.69 (s, 3H), 3.36 (t, J = 6.9 Hz, 2H), 3.05 - 2.98 (m, 1H), 2.98 - 2.91 (m, 1H), 2.62 (t, J = 8.2 Hz, 2H), 2.29 - 2.20 (m, 2H), 2.21 - 2.15 (m, 1H), 2.15 - 2.07 (m, 1H), 2.01 - 1.92 (m, 1H), 1.73 - 1.65 (m, 1H), 1.63 - 1.56 (m, 1H), 1.54 - 1.40 (m, 6H), 1.39 - 1.32 (m, 1H), 1.20 - 1.15 (m, 2H), 0.85 (d, J = 6.6 Hz, 3H), 0.82 (d, J = 6.6 Hz, 3H). 13C NMR (151 MHz, DMSO-d6) delta 172.27, 171.29, 171.06, 170.60, 158.88, 154.41, 152.69, 138.92, 136.66, 135.96, 134.44, 131.75, 127.42, 124.87, 124.31, 118.90, 108.36, 105.63, 60.04, 57.58, 55.86, 55.47, 53.11, 37.02, 34.94, 30.38, 29.30, 27.77, 26.83, 25.79, 24.92, 22.84, 22.43, 19.26, 18.20; HRMS calc. for C49H61N7O12 [M+Na]+: 962.4270. Found: 962.4273. |
Yield | Reaction Conditions | Operation in experiment |
---|---|---|
25% | A flask was charged with 10-Boc-SN-38 (197 mg, 0.400 mmol), DMAP (146 mg, 1.20 mmol) and triphosgene (47 mg, 0.16 mmol). Dichloromethane (4 mL) was added, and after stirring for 5 min, <strong>[159857-80-4]<strong>[159857-80-4]MC-Val-Cit-PAB</strong>OH</strong> (Bioconj. Chem., 2002, 13, 855-869) (230 mg, 0.400 mmol) was added, and the solution stirred for another 4 min. The reaction mixture was immediately filtered through a short silica gel column, eluting with 15:1 dichloromethane:methanol. The eluent was concentrated in vacuo, and the remaining residue was further purified by preparative TLC (15:1 dichloromethane:methanol) to give 10? (108 mg, 0.0990 mmol, 25% yield). LCMS M/Z: 1091 [M + H]. |
Yield | Reaction Conditions | Operation in experiment |
---|---|---|
41 mg | After dissolving 100 mg of purchased 10-O-Boc-SN-38 in 10 ml of dry dichloromethane,Add 25.6 mg (1 eq) of DMAP,A solution of triphosgene in dichloromethane (62 mg of triphosgene dissolved in 2 ml of dichloromethane) was added dropwise at 0C.Di Bi,Continue to react at 0C for 12hDichloromethane is removed under reduced pressureAfter dissolving with 10 ml of dry DMF,Add 144mg mc-vc-PABOH,Stir to room temperature and stir for 24h.After the preparation of liquid phase separation mc-vc-PAB-SN-38 41mg,The two-step yield is a total of 19.7%. | |
41 mg | After 100 mg of purchased 10-O-Boc-SN-38 was dissolved in 10 ml of dry dichloromethane, 25.6 mg (1 eq) of DMAP was added.A solution of triphosgene in dichloromethane (62 mg of triphosgene dissolved in 2 ml of dichloromethane) was added dropwise at 0 C.Continue to react at 0 C for 12 h,The dichloromethane was removed under reduced pressure.After dissolving in 10 ml of dry DMF,Add 144 mg mc-vc-PABOH,Stir to room temperature and stir for 24 hours.After preparative liquid phase separation, mc-vc-PAB-SN-38 41mg was obtained.The two-step yield is 19.7%. |
Yield | Reaction Conditions | Operation in experiment |
---|---|---|
77.64% | Under the room temperature condition, weighing compound d (13.91 g, 25.1 mmol), EDCI (9.62 g, 50.2 mmol), DMAP (153 mg, 1 . 26 mmol) dissolved in dichloromethane (250 ml) in, measuring the DIEA (13.1 ml, 75.3 mmol), is added to the above solution in, stir at room temperature 10 min. Weighing MC - Val - Cit - PAB linker (28.75 g, 50.2 mmol) dissolved in DMF (40 ml) in, then dropwise added to the reaction solution, after material feeding is finished, the reaction is stirred at room temperature overnight. After the reaction, to the round bottom flask is added to a solution of saturated ammonium chloride (100 ml), stirring 5 min. Extraction, collection of the organic layer, the organic layer in order to saturated salt is washed with water (100 ml × 2), then the anhydrous magnesium sulfate drying 2 h. Filter, collecting the filtrate, the filtrate is concentrated to obtain pale yellow solid material, the solid material by column chromatography purification (200 - 300 mesh silica gel, methanol: methylene chloride=5:3 as eluents), to obtain compound e (white solid material) 21.58 g, yield 77.64%. |
Yield | Reaction Conditions | Operation in experiment |
---|---|---|
82.5% | Under the room temperature condition, weighing compound a (17.14 g, 30.2 mmol), 1 - (3 - dimethylamino propyl) -3 - ethyl carbodiimide hydrochloride (11.58 g, 60.4 mmol), 4 - dimethylamino pyridine (185 mg, 1 . 51 mmol) dissolved in dichloromethane (300 ml) in, measuring the N, N - diisopropyl ethylamine (15.8 ml, 90.6 mmol), is added to the above solution in, stir at room temperature 10 min. Weighing MC - Val - Cit - PAB linker (36.59 g, 60.4 mmol) dissolved in dimethyl formamide (50 ml) in, then dropwise added to the reaction solution, after material feeding is finished, the reaction is stirred at room temperature overnight. After the reaction, to the round bottom flask is added to a solution of saturated ammonium chloride (100 ml), stirring 5 min. Collecting the organic layer, the organic layer in order to saturated salt is washed with water (100 ml × 2), then the anhydrous magnesium sulfate drying 2 h. Filter, collecting the filtrate, the filtrate is concentrated to obtain pale yellow solid material, the solid material by column chromatography purification (200 - 300 mesh silica gel, methanol: methylene chloride=2:1 as eluents), to obtain compound b (white solid material) 27.9 g, yield 82.5%. |
Yield | Reaction Conditions | Operation in experiment |
---|---|---|
48.4% | To the solution of triphosgene (26.5 mg, 0.090 mmol) in DCM (5.0 mL) was added a solution of A3 (210 mg, 0.200 mmol) and triethylamine (60.28 mg, 0.60 mmol) in DCM (5.0mL) at 0C. The reaction mixture was stirred at 15C for 30 mm. To the above mixture was added a solution of <strong>[159857-80-4]MC-VC-PAB</strong> (166.0 mg, 0.290 mmol) and triethylamine (59.0 mg, 0.58 mmol) in DMSO (3.0 mL) dropwise. The reaction mixture was stirred at 40C for 2 h. The mixture was diluted with DCM (30 mL) and washed with water (3 x 10 mL). The combinedorganic layer was dried, concentrated, and purified by column chromatography (0-10% MeOHin DCM) to afford A4 (160 mg, 48.4%) as a yellow oil. LCMS (5-95, AB, 1.5 mm): RT = 1.271 mi m/z = 828.6 [M/2+1]+ |
Yield | Reaction Conditions | Operation in experiment |
---|---|---|
48.4% | To the solution of triphosgene (26.5 mg, 0.090 mmol) in DCM (5.0 mL) was added a solution of A3 (210 mg, 0.200 mmol) and triethylamine (60.28 mg, 0.60 mmol) in DCM (5.0mL) at 0C. The reaction mixture was stirred at 15C for 30 mm. To the above mixture was added a solution of <strong>[159857-80-4]MC-VC-PAB</strong> (166.0 mg, 0.290 mmol) and triethylamine (59.0 mg, 0.58 mmol) in DMSO (3.0 mL) dropwise. The reaction mixture was stirred at 40C for 2 h. The mixture was diluted with DCM (30 mL) and washed with water (3 x 10 mL). The combinedorganic layer was dried, concentrated, and purified by column chromatography (0-10% MeOHin DCM) to afford A4 (160 mg, 48.4%) as a yellow oil. LCMS (5-95, AB, 1.5 mm): RT = 1.271 mi m/z = 828.6 [M/2+1]+ |
Yield | Reaction Conditions | Operation in experiment |
---|---|---|
140 mg | To a solution of triphosgene (41.6 mg, 0.140 mmol) in DCM (6.0 mL) was added a solution of compound A3 (300 mg, 0.280 mmol) and triethylamine (56.7 mg, 0.560 mmol) in DCM (5.0 mL). The mixture was stirred at 8C for 15 mm. The mixture was concentrated to give the crude product (307 mg, 99.9%) as a yellow solid, which was used for next step directly.To a solution of the crude product (300.0 mg, 0.270 mmol) in DCM (10 mL) was added a solution of MC VCPAB (156 mg, 0.270 mmol) and triethylamine (27.7 mg, 0.270 mmol) in DMF (6.0 mL). After the mixture was stirred at 8C for 12 h, it was concentrated and purified by flash column chromatography (6% MeOH in DCM) to give the product compound A4 (140 mg, 31%) as a yellow solid. LCMS (5-95, AB, 1.5 mm): RT =1.354 mi m/z=836.4 [M12+l]+ |
Yield | Reaction Conditions | Operation in experiment |
---|---|---|
140 mg | To a solution of triphosgene (41.6 mg, 0.140 mmol) in DCM (6.0 mL) was added a solution of compound A3 (300 mg, 0.280 mmol) and triethylamine (56.7 mg, 0.560 mmol) in DCM (5.0 mL). The mixture was stirred at 8C for 15 mm. The mixture was concentrated to give the crude product (307 mg, 99.9%) as a yellow solid, which was used for next step directly.To a solution of the crude product (300.0 mg, 0.270 mmol) in DCM (10 mL) was added a solution of MC VCPAB (156 mg, 0.270 mmol) and triethylamine (27.7 mg, 0.270 mmol) in DMF (6.0 mL). After the mixture was stirred at 8C for 12 h, it was concentrated and purified by flash column chromatography (6% MeOH in DCM) to give the product compound A4 (140 mg, 31%) as a yellow solid. LCMS (5-95, AB, 1.5 mm): RT =1.354 mi m/z=836.4 [M12+l]+ |
Yield | Reaction Conditions | Operation in experiment |
---|---|---|
41.8% | To a solution of triphosgene (30.2 mg, 0.100 mmol) in DCM (5.0 mL) was added asolution of compound AS (210 mg, 0.200 mmol) and triethylamine (61.7 mg, 0.610 mmol) inDCM (5.0 mL). The mixture was stirred at 8C for 30 mm. Then the mixture was added dropwise to a solution of MC VCPAB (139.8 mg, 0.240 mmol) and triethylamine (61.7 mg, 0.6 10 mmol) in DMF (5.0 mL). The mixture was stirred at 8C for 12 h. The mixture was concentrated and purified by flash column chromatography (8% MeOH in DCM) to give theproduct compound A6 (140 mg, 0.085 mmol, 41.8%) as a yellow oil. LCMS (5-95, AB,1.5mm): RT =1.248 mi m/z =816.l[M/2+l]+ |
Yield | Reaction Conditions | Operation in experiment |
---|---|---|
41.8% | To a solution of triphosgene (30.2 mg, 0.100 mmol) in DCM (5.0 mL) was added asolution of compound AS (210 mg, 0.200 mmol) and triethylamine (61.7 mg, 0.610 mmol) inDCM (5.0 mL). The mixture was stirred at 8C for 30 mm. Then the mixture was added dropwise to a solution of MC VCPAB (139.8 mg, 0.240 mmol) and triethylamine (61.7 mg, 0.6 10 mmol) in DMF (5.0 mL). The mixture was stirred at 8C for 12 h. The mixture was concentrated and purified by flash column chromatography (8% MeOH in DCM) to give theproduct compound A6 (140 mg, 0.085 mmol, 41.8%) as a yellow oil. LCMS (5-95, AB,1.5mm): RT =1.248 mi m/z =816.l[M/2+l]+ |
Yield | Reaction Conditions | Operation in experiment |
---|---|---|
30.7% | To a solution of triphosgene (46.0 mg, 0.160 mmol) in DCM (7.0 mL) was added a mixture of compound A3 (400.0 mg, 0.360 mmol) and triethylamine (37.0 mg, 0.370 mmol) in DCM (3.0 mL) at 0C underN2. After the reaction mixture was stirred at 0C for 30 mm, a solution of <strong>[159857-80-4]MC-VC-PAB</strong> (247.0 mg, 0.430 mmol) and triethylamine (73.0 mg, 0.720 mmol) inDMF (3.0 mL) was added at 20C under N2. The reaction mixture was stirred at 40C under N2 for 8 h. The mixture was concentrated and purified by column chromatography (0-6% MeOH in DCM) to afford compound A4 (190 mg, 30.7%) as a yellow solid. LCMS (5-95, AB, 1.5 mm):RT =1.308 mi m/z=854.2 [M12+1]+ |
Yield | Reaction Conditions | Operation in experiment |
---|---|---|
30.7% | To a solution of triphosgene (46.0 mg, 0.160 mmol) in DCM (7.0 mL) was added a mixture of compound A3 (400.0 mg, 0.360 mmol) and triethylamine (37.0 mg, 0.370 mmol) in DCM (3.0 mL) at 0C underN2. After the reaction mixture was stirred at 0C for 30 mm, a solution of <strong>[159857-80-4]MC-VC-PAB</strong> (247.0 mg, 0.430 mmol) and triethylamine (73.0 mg, 0.720 mmol) inDMF (3.0 mL) was added at 20C under N2. The reaction mixture was stirred at 40C under N2 for 8 h. The mixture was concentrated and purified by column chromatography (0-6% MeOH in DCM) to afford compound A4 (190 mg, 30.7%) as a yellow solid. LCMS (5-95, AB, 1.5 mm):RT =1.308 mi m/z=854.2 [M12+1]+ |
Yield | Reaction Conditions | Operation in experiment |
---|---|---|
34% | To a solution of triphosgene (53 mg, 0.18 mmol) in DCM (15 mL) was added a mixture of compound A3 (500 mg, 0.39 mmol) and triethylamine (45 mg, 0.44 mmol) in DCM (5.0 mL)at 0C under N2. The reaction mixture was stirred at 0C for 30 mm. To this reaction mixture was added a solution of MC VCPAB (308 mg, 0.54 mmol) and triethylamine (90 mg, 0.89 mmol) in DMSO (4.0 mL) at 10C under N2. The reaction mixture was stirred at 40C under N2 for 6 h. The mixture was diluted with DCM (30 mL), washed with water (2 x 15 mL) and the aqueous layer was extracted with EtOAc (2 x 20 mL). The combined organic layer was driedover Na2SO4, concentrated and purified by flash column (0-10% MeOH in DCM) to afford compound 8 (300 mg, 34% yield) as a yellow solid. LCMS (5-95, AB, 1.5 mm): RT =1.253 mi m/z=857.1 [M/2+1]+ |
Yield | Reaction Conditions | Operation in experiment |
---|---|---|
34% | To a solution of triphosgene (53 mg, 0.18 mmol) in DCM (15 mL) was added a mixture of compound A3 (500 mg, 0.39 mmol) and triethylamine (45 mg, 0.44 mmol) in DCM (5.0 mL)at 0C under N2. The reaction mixture was stirred at 0C for 30 mm. To this reaction mixture was added a solution of MC VCPAB (308 mg, 0.54 mmol) and triethylamine (90 mg, 0.89 mmol) in DMSO (4.0 mL) at 10C under N2. The reaction mixture was stirred at 40C under N2 for 6 h. The mixture was diluted with DCM (30 mL), washed with water (2 x 15 mL) and the aqueous layer was extracted with EtOAc (2 x 20 mL). The combined organic layer was driedover Na2SO4, concentrated and purified by flash column (0-10% MeOH in DCM) to afford compound 8 (300 mg, 34% yield) as a yellow solid. LCMS (5-95, AB, 1.5 mm): RT =1.253 mi m/z=857.1 [M/2+1]+ |
Yield | Reaction Conditions | Operation in experiment |
---|---|---|
19.8% | To a mixture of triphosgene (26.45 mg, 0.090 mmol) and 4 A MS (30 mg) in DCM (3.0 mL) was added a solution of compound A2 (220.0 mg, 0.220 mmol) and triethylamine (22.6 mg,0.220 mmol) in DCM (2.0 mL) at 0C. The mixture was stirred at 0C for 1 h, and concentrated. To the solution this isocyanate (225.0 mg, 0.220 mmol) in DCM (6.0 mL) was added a solution of MC VCPAB (34.59 mg, 0.060 mmol), Et3N (23 mg, 0.22 mmol) and 4A MS (30 mg) in DMF (2.0 mL) at 0C. After the mixture was stirred at 20C for 16 h, it was quenched with water. DCM (10 mL) was added, separated, and DCM phase was concentratedand purified by prep-TLC (15% MeOH in DCM, Rf= 0.4) to afford compound A3 (70 mg,19.8%) as light yellow oil. LCMS (5-95, AB, 1.5 mm): RT =1.107 mi m/z=793.9 [M12+l]+ |
Yield | Reaction Conditions | Operation in experiment |
---|---|---|
19.8% | To a mixture of triphosgene (26.45 mg, 0.090 mmol) and 4 A MS (30 mg) in DCM (3.0 mL) was added a solution of compound A2 (220.0 mg, 0.220 mmol) and triethylamine (22.6 mg,0.220 mmol) in DCM (2.0 mL) at 0C. The mixture was stirred at 0C for 1 h, and concentrated. To the solution this isocyanate (225.0 mg, 0.220 mmol) in DCM (6.0 mL) was added a solution of MC VCPAB (34.59 mg, 0.060 mmol), Et3N (23 mg, 0.22 mmol) and 4A MS (30 mg) in DMF (2.0 mL) at 0C. After the mixture was stirred at 20C for 16 h, it was quenched with water. DCM (10 mL) was added, separated, and DCM phase was concentratedand purified by prep-TLC (15% MeOH in DCM, Rf= 0.4) to afford compound A3 (70 mg,19.8%) as light yellow oil. LCMS (5-95, AB, 1.5 mm): RT =1.107 mi m/z=793.9 [M12+l]+ |
Yield | Reaction Conditions | Operation in experiment |
---|---|---|
37% | To a solution of triphosgene (97.1 mg, 0.33 mmol) in DCM (15 mL) was added a solution of Compound A3 (800.0 mg, 0.33 mmol) and Et3N (99.31 mg, 0.98 mmol) in DCM (5.0 mL). After the mixture was stirred at 30C for 30 mm, it was concentrated. To aboveresidue was added a solution of MC VCPAB (368.0 mg, 0.33 mmol) and Et3N (0.14 mL, 0.98 mmol) in DMF (15 mL). The mixture was stirred at 25C for 12 h. The mixture was concentrated and purified by flash column chromatography (5-10% MeOH in DCM) to give A4 (300 mg, 37%) as a yellow solid. LCMS (5-95, AB, 1.5 mm): RT =1.023 mi m/z=850.5 [M12+l]+ |
Yield | Reaction Conditions | Operation in experiment |
---|---|---|
6% | With benzotriazol-1-ol; O-(benzotriazol-1-yl)-N,N,N',N'-tetramethyluronium tetrafluoroborate; N-ethyl-N,N-diisopropylamine; In N,N-dimethyl-formamide; at 70℃; for 12h; | General procedure: To a solution of 1c (20 mg, 38 mumol) in DMF (2 mL) were added MC-VCPAB (12b, 0.11 g, 0.19 mmol), TBTU (63 mg, 0.19 mmol), HOBt (26 mg, 0.19 mmol) and DIPEA (41 mg, 0.19 mmol) at RT. The resulting mixture was stirred at 70 oC for 12 hours. No more product 2f was formed, which was monitored by LCMS. The reaction mixture was directly purified by prep-HPLC (method A) to give title compound 2f (2.6 mg, yield 6%) as a white solid. ESI m/z: 1085.3 (M + H)+.1H NMR (400 MHz, CDCl3) j 7.55-7.52 (m, 1H), 7.34-7.30 (m,1H), 6.67 (br s, 1H), 6.30-6.27 (m, 1H), 6.03 (br s, 1H), 5.16-5.11 (m, 2H), 5.09-4.87 (m, 2H), 4.80-4.61 (m, 1H), 4.60-4.30 (m, 2H), 3.47-3.44 (m, 1H), 3.21-3.10 (m, 1H), 3.03-2.96 (m, 2H), 2.79-2.55 (m, 4H), 2.35-1.73 (m, 28H), 1.71-1.32 (m, 15H), 1.29-1.07 (m, 4H), 0.99- 0.85 (m, 9H) ppm |
6% | With benzotriazol-1-ol; O-(benzotriazol-1-yl)-N,N,N',N'-tetramethyluronium tetrafluoroborate; N-ethyl-N,N-diisopropylamine; In N,N-dimethyl-formamide; at 70℃; for 12h; | To a solution of 1c (20 mg, 38 mumol) in DMF (2 mL) were added MC-VCPAB (12b, 0.11 g, 0.19 mmol), TBTU (63 mg, 0.19 mmol), HOBt (26 mg, 0.19 mmol) and DIPEA (41 mg, 0.19 mmol) at RT. The resulting mixture was stirred at 70 C for 12 hours. No more product 2f was formed, which was monitored by LCMS. The reaction mixture was directly purified by prep- HPLC (method A) to give title compound 2f (2.6 mg, yield 6%) as a white solid. ESI m/z: 1085.3 (M + H)+.1H NMR (400 MHz, CDCl3) t 7.55-7.52 (m, 1H), 7.34-7.30 (m,1H), 6.67 (br s, 1H), 6.30-6.27 (m, 1H), 6.03 (br s, 1H), 5.16-5.11 (m, 2H), 5.09-4.87 (m, 2H), 4.80-4.61 (m, 1H), 4.60-4.30 (m, 2H), 3.47-3.44 (m, 1H), 3.21-3.10 (m, 1H), 3.03-2.96 (m, 2H), 2.79-2.55 (m, 4H), 2.35-1.73 (m, 28H), 1.71-1.32 (m, 15H), 1.29-1.07 (m, 4H), 0.99-0.85 (m, 9H) ppm. |
Yield | Reaction Conditions | Operation in experiment |
---|---|---|
44% | With sodium hydrogencarbonate; In water; acetonitrile; at 4 - 20℃; for 16h;pH 8.0; | A solution of R18b (700 mg, 1.47 mmol, 1.0 eq) in water (8 mL) was diluted with 2 mL of aqueous NaHCO3 solution at 4 C and the mixture (pH = 8.0) was treated with commercially available 2,5-dioxopyrrolidin-1-yl 6-(2,5-dioxo-2,5-dihydro-1H-pyrrol-1-yl)hexanoate (408 mg, 0.9 eq) in 10 mL of acetonitrile. The suspension was stirred at room temperature for 16 h until the reaction was complete. The crude product was concentrated under reduced pressure and diluted with DMSO (5 mL). The crude product was purified by an ISCO 150g C18 column (eluents: 10 - 95% MeCN in water, 0.05 % in AcOH). Pure fractions were combined and lyophilized to afford 368 mg (44%) of compound R18 572.30; found 573.6 (M+H), (2M+H), 1145.9.1H NMR (500 MHz; DMSO-d6): ' 9.89 (s, 1H), 8.05 (d, J = 7.33 Hz, 1H), 7.81 (d, J = 8.79 Hz, 1H), 7.52 - 7.57 (m, J = 8.79 Hz, 2H), 7.21 - 7.26 (m, J = 8.79 Hz, 2H), 6.99 - 7.02 (m, 2H), 5.98 (br. s., 1H), 5.40 (s, 2H), 5.10 (t, J = 5.62 Hz, 1H), 4.35 - 4.45 (m, 3H), 4.18 (dd, J = 6.84, 8.30 Hz, 1H), 3.26 - 3.33 (m, 2H), 2.91 - 3.06 (m, 2H), 2.08 - 2.22 (m, 2H), 1.93 - 2.01 (m, 1H), 1.66 - 1.74 (m, 1H), 1.59 (dd, J = 4.40, 9.28 Hz, 1H), 1.43 - 1.55 (m, 5H), 1.32 - 1.43 (m, 1H), 1.19 (quin, J = 7.57 Hz, 2H), 0.84 (d, J = 8.30 Hz, 3H), 0.80 - 0.89 (m, 3H). |
Yield | Reaction Conditions | Operation in experiment |
---|---|---|
95% | In N,N-dimethyl-formamide; at 20℃; | Compound 6 (400 mg, 1.05 mmol) was combined with 6- maleimidohexannoic acid pentylfluorophenyl ester, MHPf (437 mg, 1.15 mmol) and DMF (10 mL) was added. The resulting mixture was stirred at room temperature overnight. (1049) [0562] The solvent was evaporated from the reaction mixture to leave a residue. The residue was titurated with ether and the resulting slid was filtered. The solid was washed with ethyl ether and dried under vacuum to give a solid in 570 mg for a 95% % yield. MS (ESI): m/e 572.66, (Mi l) , 573, (M + Na)+, 595. |
Yield | Reaction Conditions | Operation in experiment |
---|---|---|
96% | With thionyl chloride; N,N-dimethyl-formamide; at 0 - 80℃; for 0.75h; | Compound 7 (800 mg, 1.39 mmol) was combined with DMF (4 mL) and heated at 80 C until the solution was homogeneous. After 15 minutes, the reaction mixture was allowed to cool to room temperature and was subsequently cooled to 0 C. To this solution was added thionyl chloride (113 pL, 1.54 mmol) dropwise. The reaction mixture wns stirred for 30 minutes at 0 C, and then slowly quenched with ice cold water to form a solid, which was filtered and collected 380 mg. The filtrate was evaporated to dryness and titurated with ethyl ether to give an additional 400 rng of product for a 96% yield MS (ESI): m/e 590 3 (Mi l) . 591, 592. |
Tags: 159857-80-4 synthesis path| 159857-80-4 SDS| 159857-80-4 COA| 159857-80-4 purity| 159857-80-4 application| 159857-80-4 NMR| 159857-80-4 COA| 159857-80-4 structure
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P371 + P380 + P375 | In case of major fire and large quantities: Evacuate area. Fight fire remotely due to the risk of explosion. |
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Code | Phrase |
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Code | Phrase |
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Health hazards | |
Code | Phrase |
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H303 | May be harmful if swallowed |
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H305 | May be harmful if swallowed and enters airways |
H310 | Fatal in contact with skin |
H311 | Toxic in contact with skin |
H312 | Harmful in contact with skin |
H313 | May be harmful in contact with skin |
H314 | Causes severe skin burns and eye damage |
H315 | Causes skin irritation |
H316 | Causes mild skin irritation |
H317 | May cause an allergic skin reaction |
H318 | Causes serious eye damage |
H319 | Causes serious eye irritation |
H320 | Causes eye irritation |
H330 | Fatal if inhaled |
H331 | Toxic if inhaled |
H332 | Harmful if inhaled |
H333 | May be harmful if inhaled |
H334 | May cause allergy or asthma symptoms or breathing difficulties if inhaled |
H335 | May cause respiratory irritation |
H336 | May cause drowsiness or dizziness |
H340 | May cause genetic defects |
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H351 | Suspected of causing cancer |
H360 | May damage fertility or the unborn child |
H361 | Suspected of damaging fertility or the unborn child |
H361d | Suspected of damaging the unborn child |
H362 | May cause harm to breast-fed children |
H370 | Causes damage to organs |
H371 | May cause damage to organs |
H372 | Causes damage to organs through prolonged or repeated exposure |
H373 | May cause damage to organs through prolonged or repeated exposure |
Environmental hazards | |
Code | Phrase |
H400 | Very toxic to aquatic life |
H401 | Toxic to aquatic life |
H402 | Harmful to aquatic life |
H410 | Very toxic to aquatic life with long-lasting effects |
H411 | Toxic to aquatic life with long-lasting effects |
H412 | Harmful to aquatic life with long-lasting effects |
H413 | May cause long-lasting harmful effects to aquatic life |
H420 | Harms public health and the environment by destroying ozone in the upper atmosphere |
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