Fmoc Solid-phase Synthesis Of Neurotensin

Neurotensin(NT) is a kind of bioactive peptide which composed of 13 amino acids.It has multi-functions,especially on pain relieving and blood hypertension.Thus it has broad applications as potential nerve medicines in the field of medicial area.In this thesis,designed of bioreactor of solid-phase peptide synthesis and optimized of synthesis technology are focused on.Firstly,a kind of vivrational mixed bioreactor had been designed and chosen as the NT synthesis reactor.Then,NT had been synthesized successfully from C-terminal to N-terminal according to the amino acids sequence Pyr-Leu-Tyr-Glu-Gln-Lys-Pro -Arg-Arg-Pro-Tyr-Ile-Leu-OH with the raw material of FMOC-amino acids and Wang resin as the vector.The influences of combination,deblock and cleavage reagents on the finall result of synthesis were discussed.The optimization technology of NT synthesis were as follow:using DCM as the coupling solvent;TBTU/HOBt/DIEA as the coupling reagent;TFA/H₂O/thioanisole/Phenol/EDT (82:5:5:6:2) as the cleavage condition,the yield of thymopentin was 61.8%under 2 times amino acids at a coupling rate of 3～12 hours.This solid-phase synthesis method was simple and convenient,and easy to be amplified.Finally,an HPLC method was built up to analyze the crude Neurotensin.Behaviors under different HPLC conditions were studied,which proved that the resolution was so sensitive to acetonitrile concentration that the initial concentration of acetonitrile can affect the result of analysis. It was identified by HPLC/MS and the molecular weight of NT was 1672.96.The result was the same as its theoretical value.