Engineering the proteolytic specificity of activated protein C improves its pharmacological properties

被引:47
作者
Berg, DT
Gerlitz, B
Shang, J
Smith, T
Santa, P
Richardson, MA
Kurz, KD
Grinnell, BW
Mace, K
Jones, BE
机构
[1] Lilly Corp Ctr, Lilly Res Labs, Div Biores Technol & Prot, Indianapolis, IN 46285 USA
[2] Lilly Corp Ctr, Lilly Res Labs, Div Cardiovasc Res, Indianapolis, IN 46285 USA
[3] Lilly Corp Ctr, Lilly Res Labs, Div Drug Disposit, Indianapolis, IN 46285 USA
关键词
D O I
10.1073/pnas.0736918100
中图分类号
O [数理科学和化学]; P [天文学、地球科学]; Q [生物科学]; N [自然科学总论];
学科分类号
07 ; 0710 ; 09 ;
摘要
Human activated protein C (APC) is an antithrombotic, antiinflammatory serine protease that plays a central role in vascular homeostasis, and activated recombinant protein C, drotrecogin alfa (activated), has been shown to reduce mortality in patients with severe sepsis. Similar to other serine proteases, functional APC levels are regulated by the serine protease inhibitor family of proteins including alpha(1)-antitrypsin and protein C inhibitor. Using APC-substrate modeling, we designed and produced a number of derivatives with the goal of altering the proteolytic specificity of APC such that the variants exhibited resistance to inactivation by protein C inhibitor and alpha(1)-antitrypsin yet maintained their primary anticoagulant activity. Substitutions at Leu-194 were of particular interest, because they exhibited 4- to 6-fold reductions in the rate of inactivation in human plasma and substantially increased pharmacokinetic profiles compared with wild-type APC. This was achieved with minimal impairment of the anticoagulant/antithrombotic activity of APC. These data demonstrate the ability to selectively modulate substrate specificity and subsequently affect in vivo performance and suggest therapeutic opportunities for the use of protein C derivatives in disease states with elevated serine protease inhibitor levels.
引用
收藏
页码:4423 / 4428
页数:6
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