Recombinant protein and fragments thereof, method for producing said recombinant protein, synthetic gene and use of sculptin or recombinant protein for preparing a medicament or pharmaceutical composition for the prophylaxis and/or treatment of thromboembolic diseases or as a direct and specific inhibitor of thrombin

Inventors

Chudzinski-Tavassi, Ana Marisa • Goldfeder, Mauricio Barbugiani • Iqbal, Asif

Assignees

Fundacao Butantan

Abstract

A class of proteins that inhibit thrombin, particularly direct inhibitor of thrombin modified from sculptin, identified in the transcriptomics analysis of the salivary glands of ticks, as well as fragments and recombinant protein thereof, which can be used as anticoagulant agents and for the prophylaxis and/or treatment of thromboembolic diseases. These proteins fall within the fields of biochemistry, molecular biology, genetics, pharmacy and medicinal chemistry, being related to biochemical and metabolic processes.

Core Innovation

The invention relates to a novel class of tick-salivary “sculptin”-derived recombinant thrombin inhibitors and fragments. A recombinant protein comprising one sequence selected from SEQ ID NO: 1 to SEQ ID NO: 16 is described as a direct, specific, competing and reversible inhibitor of thrombin.

The recombinant thrombin inhibitors are reported to show selective thrombin inhibition versus other serine proteases, with tight-binding kinetics. The document reports direct, specific, competing and reversible thrombin inhibition, including IC50 and Ki values, and describes thrombin inhibition kinetics using inhibition and binding parameters.

The document also describes controlled proteolysis by thrombin and/or factor Xa that produces fragments. These thrombin-derived fragments retain thrombin-inhibitory activity, and functional anticoagulant effects are reported in human plasma, including prolongation measures.

The recombinant inhibitors are described in the context of therapeutic or prophylactic use, including use as a direct and specific inhibitor of thrombin for thromboembolic diseases. The document further outlines protein/gene engineering aspects, including a synthetic gene with identity thresholds to SEQ ID NO: 17, expression vector/plasmid, and recombinant expression described in E. coli.

Claims Coverage

The patent document provides two independent claims, both directed to methods of prophylaxis and/or treatment of thromboembolic diseases using an effective dose of a pharmaceutical composition comprising a recombinant protein defined by SEQ ID NOs 1–16. A second independent claim additionally frames the same recombinant-protein administration as use as a direct and specific thrombin inhibitor.

Across both independent claims, the inventive scope is defined by administering an effective dose of a pharmaceutical composition containing a recombinant protein sequence selected from SEQ ID NOs 1–16 for prophylaxis and/or treatment of thromboembolic diseases, and in the second claim also for use as a direct and specific thrombin inhibitor.

Stated Advantages

Documented Applications