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Study on the Synthesis of RGD Tripeptide by the Combination of Chemical and Enzymatic Method

Author: CaoJianJun
Tutor: ShenShuBao;OuYangPingKai
School: Nanjing University of Technology
Course: Biochemical Engineering
Keywords: RGD Peptide synthesis DCC Protease Porcine pancreatic lipase
CLC: TQ28
Type: Master's thesis
Year: 2003
Downloads: 261
Quote: 0
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Abstract


In recent years, the rapid development of genetic engineering, protein engineering and enzyme engineering technology, as well as the modernization of the separation, purification and detection means, greatly accelerate the rate of discovery of new active peptides. Research and development having a biological activity or the pharmaceutically active peptide, including the synthesis of the peptide fragment of its precursor, the international tech fields has become an important part in the competition. Traditional peptide synthesis by liquid phase chemical synthesis, commonly used include carbodiimide (DCC) method, mixed anhydride method, acid chloride method, azides method, etc., to provide a variety of options for the synthesis of a polypeptide. Later in the liquid phase chemical synthesis of peptides on the basis of further development of the solid phase synthesis method. The past three decades enzymatic peptide synthesis has become a favorite of peptide synthesis field has matured with the non-aqueous enzymology-depth research and development, application of enzymes in organic synthesis, enzymatic synthesis method compared with chemical synthesis Law shows many advantages. In recent years proteolytic enzymes, the use of a variety of sources, synthesized in a non-aqueous medium the various functions of the peptide or a precursor thereof, including some of the nutritional function of dipeptide and tripeptides, such as low-calorie high-intensity sweeteners dipeptide, RGD tripeptides and the like. Using an enzyme reactor, continuous synthesis of some of the functional peptide is close to the production scale. RGD is composed of arginine (Arg), glycine (Gly), aspartic acid (Asp) three amino sequence peptide is found in recent years, one of the short peptide sequence having biological activity. RGD sequence peptide is able to competitive inhibition the including fibrin Fib, including a variety of adhesion proteins binding to platelets, thereby inhibiting the binding of the FIB and platelet, so that in future clinical applications, and has good prospects. Identify it in the endothelial cells, antithrombotic, anticoagulant, inhibiting tumor ophthalmic diagnosis and treatment, and the treatment of burns and skin ulcers has an important role. The RGD peptide synthesis has increasingly shown its importance and potential of the market value. The purpose of this paper is a preliminary study RGD peptide synthesis process and route of liquid phase chemical method (DCC method, mixed acid anhydride method, aminolysis replaced) synthesis GD dipeptide, a variety of the RGD tripeptide experimental process condensing agent, into the mechanism of action of the anhydride reagent, and to study a variety of factors influence the extent to which the peptide synthesis, and the accumulation of data, so as to provide a theoretical basis and necessary technical constant RGD tripeptide scale preparation. In this paper, the DCC method, mixed acid anhydride method Synthesis of HCl · Gly-Asp (OBZL) 2 the dipeptide product and Z-Arg (Tos)-Gly-Asp (OBZL) 2 tripeptide product (ammonia solution to replace chloride synthesis) synthesis of the dipeptide Gly-Asp (NH 2 ) 2 product; study more of the factors including reaction time, the kind of solvent, the ratio of the substrate, and an alkali added in an amount of the chemical synthesis of the RGD sequence of the peptide. The added amount of the base specific means of triethylamine and N-methyl morpholine, and the purpose and role of the different reaction of the same. Nanjing a Master thesis X external experiment is too flat objects small attempt to not more than the pig pudsy lactone diol (N. 1.) And iodine the egg [the enzyme (lryp advised to ;) sub w this wide into eleven j '( Z 8 I · 1 Liao. Magic Gly eighteen ** HN / * and * eighteen * to \Bin. blanket M home Jin lower primary discussion cloth point he vermis M the reagents [·, the impact of factors such as the amount of water added to the enzymatic reaction. Mountains in the the laboratory hardware limitations plus * limited time T is * Mountain effect, our people. has not been excellent co-enzymatic peptide synthesis is far longer hair sticking out at our factory sub point ... servant a, the original meaning of the experimental results policy preliminary theoretical analysis and make a synthesis of RGD chemical method have a reasonable explanation, some instructive conclusions. door) ... dipeptide, tripeptide experiment is small, taking into account the low dielectric constant solvent not small), the Secretary Shing NJ fiscal from compliance w] eve {strict two objects whether eve

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