Synthesis Process Control And Separation Conditions Optimization Of Non-T Cell Peptide

If T lymphocytes are activated,they will lead to the increasingly release of cytokines,immunoglobulins,chemokines and other inflammatory mediators,which will cause synovial hyperplasia,destruction of cartilage and bone,pathological changes of chronic disabling.Non-T cell binding peptide(NTAP)is an innovative drug which is developed for this kind of disease.Compared with traditional drug,non-T cell binding peptide take effect in the pathogenesis of the disease.At present,the solid-phase peptide synthesis of NTAP process route has been completed,solution-phase peptide synthesis for NTAP is in progress.Therefore,the quality control of solid-phase synthesis of non T-cells peptide,the impurities determination and the assay method of NTAP has great significance to the safety and effectiveness of this drug.At the same time,the process monitoring of the synthesis reaction of the non-T cell binding peptide in solution-phase peptide synthesis reaction is an important part of successful completion of the liquid phase synthesis route.This thesis is mainly for the quality control of non-T cell binding peptide from solid phase synthesis and the process control of non-T cell binding peptide from solution-phase peptide synthesis.The paper consists of four parts as follows:1)The review of non T cells peptide(NTAP),including the non T cells peptide structure information,the synthesis about peptide and the separation method of peptides.2)The quality study for solid-phase peptide synthesis(SPPS)of NTAP was done.The physical and chemical properties of NTAP from SPPS was determined.The amino acid composition determination,the amino acid sequencing analysis 、 liquid chromatography-mass spectrometry was used to proof the structure.The three main impurities were assured by separation and concentration.3)The Influencing factors for separation was investigated and then the HPLC method to determine the NTAP was established.The method validation results show that the proposed method has the property of better separation,good accuracy,precision and linearity.The method was used to determine the NTAP content of different batches by external reference method with satisfied results.4)In the process of solution-phase peptide synthesis,in order to control the final product,the process of the synthesis was studied,including the detection method of amino acid raw materials and intermediates fragments by TLC and HPLC,LC-MS.The methodscan well control the purity of all the fragments,which can assure the quality of NTAP from solution-phase peptide synthesis.