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Design and evaluation of a thrombin-activable plasminogen activator
W P Yang1, J Goldstein, R Procyk
1Department of Macromolecular Biochemistry, Bristol-Myers Squibb Pharmaceutical Research Institute, Princeton, New Jersey 08543-4000.
Biochemistry
|March 1, 1994
Summary
A novel thrombin-activable plasminogen activator, 59D8-scuPA-T, selectively dissolves fresh blood clots. This targeted approach offers potential as a new antithrombotic agent for treating thrombosis.
Area of Science:
- Biochemistry
- Molecular Biology
- Pharmacology
Background:
- Developing targeted thrombolytic agents is crucial for effective clot dissolution.
- Plasminogen activators are key in breaking down fibrin clots.
- Current agents may lack specificity, leading to off-target effects.
Purpose of the Study:
- To design and characterize a novel chimeric plasminogen activator, 59D8-scuPA-T.
- To evaluate its selective activation by thrombin and fibrin binding.
- To assess its efficacy in thrombolysis of fresh clots.
Main Methods:
- Constructed a chimeric molecule fusing an anti-fibrin antibody fragment (59D8) with a thrombin-activable single-chain urokinase plasminogen activator (scuPA-T).
- Utilized site-directed mutagenesis to create the thrombin-activable scuPA-T variant.
- Performed in vitro clot lysis assays using plasma clots and assessed inhibition by heparin and hirudin.
Main Results:
- The chimeric molecule 59D8-scuPA-T demonstrated specific activation by thrombin, not plasmin.
- 59D8-scuPA-T effectively lysed thrombin-induced plasma clots in vitro.
- The agent selectively targeted and lysed fresh clots within a thrombin-induced clot model.
Conclusions:
- 59D8-scuPA-T is a potent, thrombin-activable plasminogen activator with high fibrin affinity.
- It offers selective thrombolysis of thrombin-rich clots, distinguishing them from aged clots.
- This targeted approach suggests potential as an antithrombotic agent.