Molecular design and characterization of recombinant long half-life mutants of human tissue factor pathway inhibitor

Hao Bai, Duan Ma, Yu Gao Zhang, Nong Zhang, De Sheng Kong, Hong Shen Guo, Wei Mo, Qi Qun Tang, Hou Yan Song

Research output: Contribution to journalArticlepeer-review

Abstract

Tissue factor pathway inhibitor (TFPI) is a physiological inhibitor of extrinsic pathway of coagulation and has biological functions of anticoagulation and anti-inflammation. Although TFPI has been proved to be a good therapeutic agent of sepsis, inflammatory shock, and DIC, the clinical application and therapeutic effects of TFPI are impeded because of its short half-life in vivo. In order to prolong the half-life of TFPI, homology modeling and molecule docking were performed on a computer workstation principally in protein structural biology and binding characteristics between TFPI and its receptor LRP (low-density lipoprotein receptor related protein). Two recombinant long half-life human TFPI mutants coined TFPI-Mut1 and TFPI-Mut4 were designed and expressed in E.coli. In comparison with the wild-type TFPI, TFPI-Mut1 and TFPI-Mut4 presented a few of changes in spatial configuration and a decrease in relative Gibbs free energy of docking complex by 17.3% and 21.5%, respectively, as indicated by a computer simulation.After refolding and purification, anticoagulant activities, anti-TF/FVIIa and anti-FXa activities of the mutants were found to be the same as those of wide-type TFPI. The pharmacokinetics research indicated that alpha phase half-life (t1/2α) of TFPI-Mut1 and TFPI-Mut4 were prolonged 1.33-fold and 1.96-fold respectively, beta phase half-life (t 1/2 β) of TFPI-Mut1 and TFPI-Mut4 were prolonged 1.62-fold and 4.22-fold respectively. These results suggested that TFPI-Mut1 and TFPI-Mut4 maintained the bioactivities of wild-type TFPI, prolonged half-life in vivo simultaneously and were expected for better clinical value and therapeutic effect.

Original languageEnglish (US)
Pages (from-to)1055-1060
Number of pages6
JournalThrombosis and Haemostasis
Volume93
Issue number6
DOIs
StatePublished - Jun 1 2005

Keywords

  • Half-life
  • Mutants
  • Pharmacokinetics
  • TFPI

ASJC Scopus subject areas

  • Hematology

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