Amerikanische Gesellschaft für Hirudotherapie

Improvement of the specificity of dipetarudin by site directed mutagenesis

Research article published in Thrombosis and haemostasis (2005)

Zuletzt aktualisiert: June 18, 2026Geprüft von: ASH Editorial Board
Research article — evidence reviewArticle reference
Evidence: Research reportArzneimittelentwicklungKlinische StudienSpeichel-PharmakologieLpez M et al. · Thrombosis and haemostasis, 2005

Abstract

Protease specificity is crucial to the design of thrombin inhibitors as inhibition of other physiologically relevant serine-proteases can compromise their clinical use. Dipetarudin, a potent thrombin inhibitor, also inhibits trypsin and plasmin. Due to the specificity of an inhibitor being influenced by the amino acid residue at the P1 position, we replaced the Arg10 at P1 position of dipetarudin by a histidine, which is the P1 residue of rhodniin, a very specific thrombin inhibitor. The amino acid replacement was carried out by site directed mutagenesis. The mutant, dipetarudinR10H, showed a loss of plasmin and trypsin inhibitory activities present in its wild-type counterpart and a 3-fold higher dissociation constant for thrombin than dipetarudin. However, compared to dipetarudin and r-hirudin, dipetarudinR10H showed similar activity in coagulation screening assays such as activated partial thromboplastin time (aPTT), prothrombin time (PT), ecarin clotting time (ECT) and ecarin chromogenic assay (ECA).

Abstract sourced from PubMed (NCBI) for the cited record. See the original publication for the authoritative version.

Publication typeJournal ArticleResearch Support, Non-U.S. Gov't
Indexed MeSH termsAmino Acid SubstitutionBlood CoagulationBlood Coagulation TestsHumansMutagenesis, Site-DirectedRecombinant Fusion ProteinsSerine Proteinase InhibitorsStructure-Activity RelationshipSubstrate SpecificityThrombin

Zusammenfassung

Protease specificity is crucial to the design of thrombin inhibitors as inhibition of other physiologically relevant serine-proteases can compromise their clinical use.

Warum dies für die Hirudotherapie relevant ist

This study used site-directed mutagenesis to replace the P1 arginine at position 10 of dipetarudin with histidine, yielding a mutant (dipetarudinR10H) that lost plasmin and trypsin inhibitory activity. In coagulation screening assays, including activated partial thromboplastin time and prothrombin time, dipetarudinR10H showed similar activity to both dipetarudin and r-hirudin. The work is relevant because it evaluates engineered thrombin inhibitors and directly compares their functional coagulation effects against r-hirudin. However, the abstract does not identify the biological origins of r-hirudin or dipetarudin, nor does it involve live leeches. Therefore, the connection to the leech secretome is entirely indirect, relying solely on the in-vitro comparison with r-hirudin.

Zitation

Improvement of the specificity of dipetarudin by site directed mutagenesis

Lpez M et al. · Thrombosis and haemostasis, 2005

DOI

Verwandter klinischer Kontext

Erfahren Sie, wie diese Forschung mit der klinischen Praxis verknüpft ist

Drug PipelineDrug DevelopmentDirect Thrombin InhibitorsClinical EvidenceClinical ApplicationsSalivary Gland SecretionAnti-Inflammatory MechanismsSalivary ComplexHemostasisHirudinOtolaryngology

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