Solid-phase Synthesis And Purification For Thymopentin & Fusukang

Solid-Phase Synthesis and purification of thymopentin were studied in this thesis. By studying the effects of coupling time, coupling solvents, coupling reagents and cleavage conditions on the yield of thymopentin systemically during Fmoc-based solid-phase peptide synthesis, the protocol was optimized. Using DMF as the coupling solvent; HATU as the coupling reagent; TFA, p-methyl phenol, EDT and H2O (97:0.25:0.25:2.5) as the cleavage condition, the yield of thymopentin was 79.68% and the purity was 80.36% under 2 equiv. amino acid at a coupling rate of 3 h per amino acid. The Fmoc-amino quantified by RP-HPLC was evaluated the reaction extent. Crude thymopentin was isolated and purified, and the optimal technology was established: The crude product was purified by ion-exchange chromatography (IEC), and was completely purified by reversed-phase high performance liquid chromatography (RP-HPLC) again. The total yield was 68% with the purity of 98%. In addition, Solid-Phase Synthesis and purification of Fusukang were studied. By studying the effects of coupling reagents, coupling time and washing manner on yield during Fmoc-based solid-phase synthesis, the technique was optimized. Using HATU as the coupling regent, washing 5 times with DCM and reacting with optimized coupling time of each amino acid, the yield of Fusukang was 21.02% and the purity was 32.30%. According to isolation and purification of Fusukang, the optimal technology was established: The crude product was purified by ion-exchange chromatography (IEC), and was completely purified by reversed-phase high performance liquid chromatography (RP-HPLC), the total yield was33% with the purity of 95%.