The Preparation And Pharmacodynamic Studies Of Glucose Regulation Of Long-term Glucose Regulation Polypeptide

DM is a serious diseases that threat human health and life,the incidence and mortality rate is rising year by year.95%of the patients are T2DM.T2DM is caused by insulin resistance and insulin deficiency,also called Non-insulin-dependent diabetes mellitus.Many patients who have T2DM still use insulin treatment,this causes the insulin dosage increasing and hypoglycemic effect is getting worse,which urges researchers to continue to develop the non-insulin drug.Because the non-insulin drug isn't naturally occurring bioactive molecules in vivo.The long-term using can lead to varieties of adverse reactions,and will seriously limit the application of this kind of drug.Currently,the main drugs for the treatment of T2DM are still based on durgs like Sulfonylurea,Biguanide,which promoting the insulin secretion.But except for the common adverse reactions,the most obvious drawback of this kind of drugs may cause persistent hypoglycemia,and long-term using brings on the oversecretion of insulin and finally causes the failure of Beta Cell.In addition,because the traditional treatment of T2DM particularly emphasizes on ameliorating high blood glucose status,this kind of treatment is only palliative,causing the weakening of glucose control,and the decline of the number and function of Beta Cell of most patients gradually.Therefore,developing new drugs that can control T2DM and varieties of complications in a safer and more effective way,especially endogenous substances or their analogues,which regulate blood glucose status,is the hotspot and focus of current research.GLP-1 is a kind of important endocrine hormone of intestinal tract that possess the abilities of decreasing blood glucose status,enhancing insulin secretion,proliferation ability on Bate Cell,decreasing the glucagon secretion,ameliorating Insulin Resistance,delaying gastric emptying,controlling cravings,protecting the cardiovascular and nervous system,ect,which have several features of ideal hypoglycemics,and have become the hotspot of research and development in the field of T2DM treating.But the natural GLP-1 is very easy to be degraded by DDP-IV,that limits the clinical application.Therefore,trying to prolong the action time is the point of current research.This study is based on the studies mentioned above,we did researches including high expression,fermentation,renaturation,purification,mass detection and preliminary study on pharmacodynamics about a new and combined Pro-GLP-1 analog,that was constructed by our research group.Find out the following results.1.Find out the expression conditions of pilot fermentation of 601-GLP-1 by changing series of conditions and draw the fermentation curve.2.Manage Ni²⁺-chelating chromatography of 601-GLP-1 by finding the renaturation condition,and achieve good renaturation rates and effects.3.Finish the experiment of protein purification,concentration and endotoxin elimination experimentation.Got 601-GLP-1 proteins that match the standard of endotoxin using the method of washing inclusion bodies,chromatography of anion-exchange and cation-exchange4.Finish the primary research of effect of blood glucose regulation.Hypoglycemic experiment,glucose tolerance experiment and stimulating insulin secretion experiment proved the marked effect of 601-GLP-1.