Diisopropylcarbodiimide → (S)-2-Mercapto-3-methylbutyroyl-(L)-homo-phenylalanyl-(L)-phenylalanine

Combine Fmoc-(L)-phenylalanine-NovaSyn® TGT resin (1.0 g, 0.15 mmol/g, Nova Biochem), dimethylformamide (3 mL) and 50% piperidine/dimethylformamide (3 mL). After 5 minutes, filter, add dimethylformamide (3 mL) and 50% piperidine/dimethylformamide (3 mL), bubble for 10 minutes, and filter. Rinse the resin with dimethylformamide (10 times 5 mL), dichloromethane (3 times 5 mL), and then dimethylformamide (5 mL). Kaiser (ninhydrin) test indicates complete deprotection. Add Fmoc-(L)-homophenylalanine (0.240 g, 0.5 mmol), O-(7-azabenzotriazol-1-yl)-1,1,3,3-tetramethyluronium hexafluorophosphate (0.228 g, 0.60 mmol), and N,N-diisopropylethylamine (0.21 mL, 1.20 mmol), and dimethylformamide (5 mL). After 15 hours, filter, rinse the resin with dimethylformamide (9 times 5 mL), dichloromethane (3 times 5 mL), and then dimethylformamide (5 mL). Add dimethylformamide (3 mL) and 50% piperidine/dimethylformamide (3 mL), bubble for 10 minutes, and filter. Rinse the resin with dimethylformamide (10 times 5 mL), dichloromethane (3 times 5 mL), and then dimethylformamide (5 mL). Kaiser (ninhydrin) test indicates complete deprotection. Add (R)-2-bromo-3-methylbutyric acid (0.145 g, 0.6 mmol), 1,3-diisopropylcarbodiimide (0.076 g, 0.60 mmol), and N,N-diisopropylethylamine (0.21 mL, 1.20 mmol) in N-methylpyrrolidinone (5 mL). After 15 hours, filter, rinse the resin with dimethylformamide (9 times 5 mL), dichloromethane (3 times 5 mL), and then dimethylformamide (5 mL). Combine the resin with a degassed solution of potassium thioacetate (0.137 g, 1.2 mmol) in N-methylpyrolidinone (5 mL). After 3 hours, filter and rinse the resin with dimethylformamide (9 times 5 mL). Combine the resin with a degassed saturated methanolic ammonia solution (5 mL). After 5 minutes, filter and rinse the resin with methanol (5 times 5 mL). Combine the resin and a degassed 95% trifluoroacetic acid/4% water/1% 2-mercaptoethanol solution (10 mL). After 3 hours, filter, rinse with 50% trifluoroacetic acid/dichloromethane (2 times 10 mL). Concentrate the filtrate in vacuo to give a residue. Combine the residue with 1% trifluoroacetic acid/acetonitrile solution and evaporate under a stream of nitrogen to give a residue. Combine the residue and water (about 4 mL) and lyophilize to give a residue. Combine the residue and acetonitrile and water and again lyophilize to give a residue. Purify by HPLC to give the title compound: retention time=4:24 minutes, YMC Inc. 4.0 mm×50 mm ODS-A column, flow rate 1.0 mL/minute, initial conditions 70% A (95% water/5% acetonitrile/0.1% trifluoroacetic acid) and 30% B (5% water/95% acetonitrile/0.085% trifluoracetic acid) and a linear gradient over 5 minutes to 100% B. Mass Spectrum (MH+=443).

1. filtrationfilter
2. additionadd dimethylformamide (3 mL) and 50% piperidine/dimethylformamide (3 mL)
3. custombubble for 10 minutes
4. filtrationfilter
5. washRinse the resin with dimethylformamide (10 times 5 mL), dichloromethane (3 times 5 mL)
6. waitAfter 15 hours
7. filtrationfilter
8. washrinse the resin with dimethylformamide (9 times 5 mL), dichloromethane (3 times 5 mL)
9. customAdd dimethylformamide (3 mL) and 50% piperidine/dimethylformamide (3 mL), bubble for 10 minutes
10. filtrationfilter
11. washRinse the resin with dimethylformamide (10 times 5 mL), dichloromethane (3 times 5 mL)
12. waitAfter 15 hours
13. filtrationfilter
14. washrinse the resin with dimethylformamide (9 times 5 mL), dichloromethane (3 times 5 mL)
15. waitAfter 3 hours
16. filtrationfilter
17. washrinse the resin with dimethylformamide (9 times 5 mL)
18. waitAfter 5 minutes
19. filtrationfilter
20. washrinse the resin with methanol (5 times 5 mL)
21. waitAfter 3 hours
22. filtrationfilter
23. washrinse with 50% trifluoroacetic acid/dichloromethane (2 times 10 mL)
24. concentrationConcentrate the filtrate in vacuo
25. customto give a residue
26. customevaporate under a stream of nitrogen
27. customto give a residue
28. customto give a residue
29. customto give a residue
30. customPurify by HPLC

ORDレコード

ord-5ebe91982a314058bd7c1f8ed59a5eee

出典データセット

uspto-grants-2002_03

DOI

10.6084/m9.figshare.5104873.v1

特許番号

US06352976B1