Recombinant antitrypsin Pittsburgh undergoes proteolytic cleavage during E. coli sepsis and fails to prevent the associated coagulopathy in a primate model

Thromb Haemost. 1998 Nov;80(5):816-21.

Abstract

During severe sepsis there is dramatic activation of both contact proteases and the coagulation pathway. These processes contribute to the development of shock and disseminated intravascular coagulation (DIC) respectively. The Pittsburgh mutant of antitrypsin (358Met-Arg) is a novel protease inhibitor with activity against both thrombin and the contact proteases and should therefore prove beneficial as a therapeutic agent in the management of septic shock. This hypothesis was supported by an earlier study in a pig model where recombinant antitrypsin Pittsburgh (rAT Pittsburgh) at a concentration of 1 microM alleviated some of the features of shock, but did not improve survival. In order to reduce the lethal effects of E. coli sepsis we postulated that a higher concentration of antitrypsin Pittsburgh would be necessary. To test this hypothesis we used rAT Pittsburgh in a primate model. This was chosen in preference to another species as E. coli sepsis in the primate has been well characterised and closely resembles the changes seen in man. Surprisingly this treatment did not alleviate the features of shock and unexpectedly appeared to exacerbate the associated coagulopathy. We propose two possible mechanisms for this unforeseen outcome. The first results from the broad spectrum of activity of antitrypsin Pittsburgh. As well as inhibiting thrombin and the contact proteases, the Pittsburgh mutant also inhibits activated protein C. Inhibition of the protein C system is known to exacerbate septic shock. Secondly, a significant quantity of inactive antitrypsin Pittsburgh, cleaved at the reactive centre, was detected in the plasma of the treated animals. Proteolytically altered serpins, including antitrypsin. have been shown to enhance the inflammatory process. Therefore the accumulation of cleaved rAT Pittsburgh might be expected to exacerbate septic shock.

Publication types

  • Comparative Study
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Disseminated Intravascular Coagulation / drug therapy
  • Disseminated Intravascular Coagulation / etiology
  • Disseminated Intravascular Coagulation / prevention & control*
  • Dogs / blood
  • Drug Evaluation, Preclinical
  • Endopeptidases / metabolism
  • Escherichia coli Infections / complications*
  • Female
  • Humans
  • Papio / blood
  • Partial Thromboplastin Time
  • Protease Inhibitors / pharmacokinetics*
  • Protease Inhibitors / therapeutic use
  • Protease Inhibitors / toxicity
  • Rabbits / blood
  • Recombinant Fusion Proteins / pharmacokinetics
  • Recombinant Fusion Proteins / therapeutic use
  • Recombinant Fusion Proteins / toxicity
  • Shock, Septic / complications
  • Shock, Septic / drug therapy*
  • Species Specificity
  • Swine / blood
  • Treatment Failure
  • alpha 1-Antitrypsin / genetics
  • alpha 1-Antitrypsin / pharmacokinetics*
  • alpha 1-Antitrypsin / therapeutic use
  • alpha 1-Antitrypsin / toxicity

Substances

  • Protease Inhibitors
  • Recombinant Fusion Proteins
  • alpha 1-Antitrypsin
  • alpha 1-antitrypsin Pittsburgh
  • Endopeptidases