【摘 要】
：
The purpose of this research is to design a mosaic protein which could be used to cure cephalophyma and cerebral edema during acute cerebral stroke.The foundation of designing this mosaic protein (TNH
【机 构】
：
Bioengineering College of Chongqing Institute of Technology, Chongqing400050 ;Chongqing Fagen Biomed
【出 处】
：
2005 WHTS3rd Annual Congress of International Drug Discovery
论文部分内容阅读
The purpose of this research is to design a mosaic protein which could be used to cure cephalophyma and cerebral edema during acute cerebral stroke.The foundation of designing this mosaic protein (TNHH) is combination with the C peptide of the hirudin and neutrophil inhibition factor (NIF) by inseting a hinge region of fibrin binding region, in which hirudin C-peptide can inhibit thrombin activity and NIF can inhibit neutrophil activity by specific blocking CD 1 lb/CD 18.Firstly,selecting the optimal scheme and amino acid sequence to design TNHH by protein modeling, then artificially synthesizing cDNA segment and constructing bacterial expression vector.To get high purity and active recombinant TNHH, the expressing strains were cultured in fermentor, inclusion bodies were extracted and renaturated, two-step chromatographies were performed.The characteristics of recombinant TNHH are that IC50 of inhibiting WBC is 10 nmol/L, inhibiting thrombin activity is 1000 ATU/mg, the combination ratio with fibrin is 50 nmol/L, molecular weight is 44Kd and pI is 10.5.Amino acid sequencing and secondary structure analyses (disulfide bond and CD) were compatible with theoretical design.Recombinant TNHH in rat acute cerebral infarction model and acute cerebral hemorrhage model showed that it could eliminate the areas of cerbral infarction and cerebral hemorrhage respectively, decrease cephalophyma and edema, and enhance the recovery of behaviors after brain damage.The results demonstrated TNHH is potential to be a candidate drug in the treatment of cephalophyma and cerebral edema during acute cerebral stroke.
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