Reaktion #593947
ord-385167b2418642f6bdd354b18805363b
Reaktionsgleichung
Reagenzien
Lösungsmittel
Reaktionsbedingungen
Vorschrift
After 4.44 g (0.45 mmol/g) of Rink Amide MBHA resin commercially available was swollen in DMF, the resin was treated with 50 mL of 20% piperidine/DMF solution for 20 minutes to remove the Fmoc group. The resulting resin was washed with DMF and treated in DMF with 4.21 g (8 mmol) of Fmoc-Trp(Boc)-OH, 1.27 mL (8 mmol) of DIPCDI and 1.31 g (8 mmol) of HOOBt at room temperature for 90 minutes, whereby Trp(Boc) was introduced to give the Fmoc-Trp(Boc)-Rink Amide MBHA resin. In a similar manner, Orn(Mtt) was introduced to give 2 mmol of the Fmoc-Orn(Mtt)-Trp(Boc)-Rink Amide MBHA resin. After the resin obtained was washed and swollen in toluene, 30 mL of TFA/TIS/TFE/toluene (1/5/47/47) was added, followed by shaking for 30 minutes and removing the solution by filtration. This procedure was repeated until yellow color caused by free Mtt group in a TFA/TIS/TFE/toluene (1/5/47/47) solution disappeared when the solution was added; thus the Mtt group was removed. The resulting Fmoc-Orn-Trp(Boc)-Rink Amide MBHA resin was neutralized with 5%-DIEA/toluene solution. After washing with toluene, 15 mL of toluene-TFE (4:1) and 1.95 g (6 mmol) of N-methyl-N,N′-bis-Boc-1-guanylpyrazole obtained in REFERENCE EXAMPLE 1 were added to the resin. DIEA was added to the mixture to adjust pH of the solution to 10. The solution was shaken for 15 hours to give Fmoc-Arg(Boc2,Me)-Trp(Boc)-Rink Amide MBHA resin (2 mmol). After the resin obtained was dried in MeOH, 0.03 mmol was weighed and reswollen in DMF. Leu was introduced in the same manner as described above to give 0.03 mmol of Fmoc-Leu-Arg(Boc2,Me)-Trp(Boc)-Rink Amide MBHA resin. After Fmoc deprotection in 20% piperidine/DMF solution, the resin was treated in DMF with 51.5 mg (0.12 mmol) of Fmoc-Phe-Ψ(CH2CH2)-Gly-OH obtained in REFERENCE EXAMPLE 3, 19.1 μl (0.12 mmol) of DIPCDI and 240 μl of 0.5 M HOAt/DMF solution at room temperature for 150 minutes. The resin was washed with DMF and then treated with 10.9 μl of (0.12 mmol) of Ac2O and 20.9 μl (0.12 mmol) of DIEA for capping of residual amino groups. Subsequently, the resin was shaken overnight in 2 mL of DMF solution of Ac-D-Tyr-Hyp-Asn-Thr-OH (0.06 mmol) synthesized in a conventional manner of the liquid-phase process, 31.2 mg (0.06 mmol) of PyAOP, 120 μl (0.06 mmol) of 0.5 M HOAt/DMF solution and 10.5 μl (0.06 mmol) of DIEA to give Ac-D-Tyr-Hyp-Asn-Thr-Phe-Ψ(CH2CH2)-Gly-Leu-Arg(Boc2,Me)-Trp(Boc)-Rink Amide MBHA resin. After drying, 1.5 mL of TFA/PhSMe/m-cresol/H2O/TIS/EDT (80/5/5/5/2.5/2.5) was added to the resin and stirred for 90 minutes.