Molecular dynamic and pharmacological studies on protein-engineered hirudin variants of Hirudinaria manillensis and Hirudo medicinalis
Basic science / preclinical published in British journal of pharmacology (2022)
Abstract
BACKGROUND AND PURPOSE: Hirudin variants are the most powerful thrombin inhibitors discovered to date, with a lower risk of bleeding than heparin. For anticoagulation, the C-termini of hirudin variants bind to the exocite I of thrombin. Anticoagulant effects of gene-recombinant hirudin are weaker than natural hirudin for the reason of lacking tyrosine O-sulfation at C-terminus. EXPERIMENTAL APPROACH: An integrative pharmacological study was carried out using molecular dynamic, molecular biological and in vivo and in vitro experiments to elucidate the anticoagulant effects of protein-engineered hirudins. KEY RESULTS: Molecular dynamic analysis showed that modifications of the C-termini of hirudin variant 1 of Hirudo medicinalis (HV1) and hirudin variant 2 of Hirudinaria manillensis (HM2) changed the binding energy of the C-termini to human thrombin. The study indicated that Asp61 of HM2 that corresponds to sulfated Tyr63 of HV1 is critical for inhibiting thrombin activities. Further, the anticoagulant effects of HV1 and HM2 were improved when the amino acid residues adjacent to Asp61 were mutated to Asp. These improvements were prolongation of the activated partial thromboplastin time, prothrombin time and thrombin time of human blood, and decreased Ki and IC50 values. In the in vivo experiments, mutations at C-termini of HV1 and HM2 significantly changed partial thromboplastin time, prothrombin and thrombin time CONCLUSION AND IMPLICATIONS: The study indicated that the anticoagulant effects of gene-engineered HM2 are stronger than gene-engineered HV1 and HM2-E60D-I62D has the strongest effects and could be an antithrombotic with better therapeutic effects.
Abstract sourced from PubMed (NCBI) for the cited record. See the original publication for the authoritative version.
Resumen
Hirudin variants are the most powerful thrombin inhibitors discovered to date, with a lower risk of bleeding than heparin. For anticoagulation, the C-termini of hirudin variants bind to the exocite I of thrombin.
Por qué esto importa para la hirudoterapia
Este estudio examinó cómo la ingeniería de proteínas y mutaciones específicas de aminoácidos en el C-terminal de variantes de hirudin provenientes de Hirudo medicinalis y Hirudinaria manillensis afectan su unión a la trombina humana y su actividad anticoagulante general in vitro e in vivo. Para la hirudoterapia y la aplicación clínica del secretoma salivar de las sanguijuelas, estos hallazgos son sumamente relevantes porque proporcionan información mecanística sobre cómo superar la eficacia reducida del hirudin recombinante causada por la ausencia de O-sulfatación natural de tirosina. Al identificar mutaciones específicas que mejoran la inhibición de la trombina y prolongan los tiempos de coagulación sanguínea, la investigación destaca vías para optimizar los anticoagulantes derivados naturalmente de las sanguijuelas para uso terapéutico. Como advertencia honesta, se trata de una investigación preclínica que utiliza dinámica molecular y modelos animales de laboratorio; se centra completamente en la ingeniería de proteínas recombinantes en lugar de la terapia con sanguijuelas vivas, y la seguridad o eficacia clínica de estas variantes modificadas en pacientes humanos reales permanece sin establecer.
Citación
Molecular dynamic and pharmacological studies on protein-engineered hirudin variants of Hirudinaria manillensis and Hirudo medicinalis.
Sun Y et al. · British journal of pharmacology, 2022
Contexto clínico relacionado
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Añadido a la biblioteca ASH: March 18, 2026 · Última actualización del sitio: June 18, 2026