反应详情

N-{[4-((2R,3R)-1-(4-Chlorophenyl)-3-{[2-(4-chlorophenyl)-2-oxoethyl]thio}-4-oxoazetidin-2-yl)phenoxy]acetyl}glycine (14.6 mg, 0.026 mmol) and N-methylmorpholine (20 μl, 0.18 mmol) were dissolved in DCM (2 ml). tert-Butyl ff-(tert-butoxycarbonyl)-D-lysinate hydrochloride (11.1 mg, 0.033 mmol) was added and after 5 minutes TBTU (9.8 mg, 0.031 mmol) was added to the suspension. The mixture was stirred overnight. Additional tert-butyl N6-(tert-butoxycarbonyl)-D-lysinate (4.8 mg, 0.014 mmol), N-methylmorpholine (10 μl, 91 μmol) and TBTU (4.6 mg, 0.014 mmol) were added and the mixture was stirred for 2.5 h. The formation of the ester was confirmed. M/z: 855.4 (M−H). DCM (3 ml) and water (5 ml) were added and the solution was acidified to pH 3 with KHSO4 (2M). The organic phase was washed with water (2×5 ml). The combined water phases were extracted with DCM (2×5 ml). The organic phase was dried over Na2SO4, filtered and concentrated under reduced pressure. The oily residue was dissolved in DCM (1.5 ml) and TFA (1 ml) was added. The mixture was stirred for 2.5 h. The mixture was concentrated and co-evaporation with toluene was performed to assist the removal of TFA. The residue was dissolved in methanol (2 ml) and sodium borohydride (10.4 mg, 0.027 mmol) was added. After 15 minutes, ammonium acetate buffer (0.1M, 1.5 ml) was added and the solvent was removed under reduced pressure. The residue was purified with preparative HPLC on a C8 column. A gradient from 20% to 50% MeCN in 0.1M ammonium acetate buffer was used as eluent. After lyophilisation, the title compound was obtained as a white solid (10.8 mg, 59%). H-NMR (400 MHz, DMSO-d6): 1.18-1.36 (m, 2H), 1.41-1.70 (m, 4H), 2.71 (t, 2H), 2.84-2.97 (m, 2H), 3.72-3.75 (brd, 2H), 3.93 (m, 1H), 4.30 (d, 0.5H), 4.34 (d, 0.5H), 4.52 (s, 2H), 4.68-4.77 (m, 1H), 5.03 (d, 0.5H), 5.07 (d, 0.5H), 6.98 (d, 2H), 7.20 (d, 2H), 7.31-7.38 (m, 8H), 7.63-7.72 (m, 1H), 8.34 (t, 1H). M/z: 705.1.

1. additionwas added to the suspension
2. stirringthe mixture was stirred for 2.5 h
3. washThe organic phase was washed with water (2×5 ml)
4. extractionThe combined water phases were extracted with DCM (2×5 ml)
5. dry with materialThe organic phase was dried over Na2SO4
6. filtrationfiltered
7. concentrationconcentrated under reduced pressure
8. dissolutionThe oily residue was dissolved in DCM (1.5 ml)
9. additionTFA (1 ml) was added
10. stirringThe mixture was stirred for 2.5 h
11. concentrationThe mixture was concentrated
12. customthe removal of TFA
13. dissolutionThe residue was dissolved in methanol (2 ml)
14. waitAfter 15 minutes
15. customthe solvent was removed under reduced pressure
16. customThe residue was purified with preparative HPLC on a C8 column