Tyrosine phosphorylation of recombinant hirudin increases affinity to thrombin and antithrombotic activity
Basic science / preclinical published in Proteins (2024)
Abstract
Thrombin is one of the key enzymes of the blood coagulation system and a promising target for the development of anticoagulants. One of the most specific natural thrombin inhibitors is hirudin, contained in the salivary glands of medicinal leeches. The medicinal use of recombinant hirudin is limited because of the lack of sulfation on Tyr63, resulting in a 10-fold decrease in activity compared to native (sulfated) hirudin. In the present work, a set of hirudin derivatives was tested for affinity to thrombin: phospho-Tyr63, Tyr63(carboxymethyl)Phe, and Tyr63Glu mutants, which mimic Tyr63 sulfation and Gln65Glu mutant and lysine-succinylated hirudin, which enhance the overall negative charge of hirudin, as well as sulfo-hirudin and desulfo-hirudin as references. Using steered molecular dynamics simulations with subsequent umbrella sampling, phospho-hirudin was shown to exhibit the highest affinity to thrombin among all hirudin analogs, including native sulfo-hirudin; succinylated hirudin was also prospective. Phospho-hirudin exhibited the highest antithrombotic activity in in vitro assay in human plasma. Taking into account the modern methods for obtaining phospho-hirudin and succinylated hirudin, they are prospective as anticoagulants in clinical practice.
Abstract sourced from PubMed (NCBI) for the cited record. See the original publication for the authoritative version.
Resumen
Thrombin is one of the key enzymes of the blood coagulation system and a promising target for the development of anticoagulants. One of the most specific natural thrombin inhibitors is hirudin, contained in the salivary glands of medicinal leeches.
Por qué esto importa para la hirudoterapia
Este estudio utilizó simulaciones de dinámica molecular dirigida y ensayos in vitro en plasma humano para comparar la afinidad por la trombina y la actividad antitrombótica entre derivados de hirudin recombinante—incluidos phospho-Tyr63, mutantes con modificación de carga y formas succiniladas—frente a la sulfo-hirudina nativa y la desulfo-hirudina. El trabajo es directamente relevante para la hirudoterapia y el secretoma salivar de la sanguijuela: la hirudin es el principal inhibidor de la trombina procedente de la saliva de la sanguijuela medicinal, y la caracterización de cómo modificaciones postraduccionales específicas (particularmente la fosforilación que imita la sulfatación nativa de Tyr63) potencian la unión y la actividad puede orientar el diseño de anticoagulantes recombinantes más potentes derivados de esta proteína salivar. Sin embargo, los hallazgos se limitan al modelado computacional y a ensayos in vitro en plasma sin datos clínicos en animales ni humanos, y las proyecciones de los autores sobre la utilidad clínica son prospectivas más que demostradas. Sería necesaria la validación mediante estudios in vivo y ensayos clínicos antes de poder formular cualquier afirmación terapéutica.
Citación
Tyrosine phosphorylation of recombinant hirudin increases affinity to thrombin and antithrombotic activity.
Volkova A, Semenyuk P · Proteins, 2024
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Añadido a la biblioteca ASH: March 18, 2026 · Última actualización del sitio: June 18, 2026