Screening of the Promising Direct Thrombin Inhibitors from Haematophagous Organisms. Part I: Recombinant Analogues and Their Antithrombotic Activity In Vitro.

М А Костромина ,Igor A Dyachenko,Natalia A Perepechenova,E S Sadovnikova ,А Н Мурашев ,Elena V Grigorieva,Alina M Ismailova,Yu A Palikova ,Evgeniy A Zayats,Dmitry D Lykoshin,Elena A Tukhovskaya,А П Момот ,Г А Слащева ,А Н Мамаев ,Nadezhda I Novikova,I N Kravchenko ,Elvira R Shaykhutdinova ,В А Паликов ,Yu A Abramchik ,Р С Есипов

doi:10.3390/biomedicines10010011

Abstract

The success in treatment of venous thromboembolism and acute coronary syndromes using direct thrombin inhibitors has stimulated research aimed at finding a new anticoagulant from haematophagous organisms. This study deals with the comparison between hirudin-1 from Hirudomedicinalis(desirudin), being the first-known and most well-studied natural anticoagulant, along with recombinant analogs of haemadin from the leech Haemadipsa sylvestris, variegin from the tick Amblyomma variegatum, and anophelin from Anopheles albimanus. These polypeptides were chosen due to their high specificity and affinity for thrombin, as well as their distinctive inhibitory mechanisms. We have developed a universal scheme for the biotechnological production of these recombinant peptides as pharmaceutical substances. The anticoagulant activities of these peptides were compared using the thrombin amidolytic activity assay and prolongation of coagulation time (thrombin time, prothrombin time, and activated partial thromboplastin time) in mouse and human plasma. The preliminary results obtained suggest haemadin as the closest analog of recombinant hirudin-1, the active substance of the medicinal product Iprivask (Aventis Pharmaceuticals, USA) for the prevention of deep venous thrombosis in patients undergoing elective hip or knee replacement surgery. In contrast, variegin can be regarded as a natural analog of bivalirudin (Angiomax, The Medicines Company), a synthetic hirudin-1 derivative certified for the treatment of patients undergoing percutaneous coronary intervention and of patients with unstable angina pectoris after percutaneous transluminal coronary angioplasty.

Highlights

Disturbances in the blood coagulation system due to an imbalance of coagulation, fibrinolysis, and inflammation processes lead to the formation of blood clots in the vessels.Arterial thrombosis is the major cause of ischemic stroke and acute myocardial infarction, whereas venous thrombosis can lead to deep vein thrombosis (DVT), pulmonary embolism (PE), and unstable angina [1]
We have reported the production of the recombinant hirudin-1 and anophelin, an anticoagulant from the malaria mosquito Anopheles albimanus [36,37]
We have used the original pH-dependent Cterminal cleavage construct based on the GyrA mini-intein from Mycobacterium xenopi (Figure S1b) [37]

Summary

Introduction

Disturbances in the blood coagulation system due to an imbalance of coagulation, fibrinolysis, and inflammation processes lead to the formation of blood clots in the vessels.Arterial thrombosis is the major cause of ischemic stroke and acute myocardial infarction, whereas venous thrombosis can lead to deep vein thrombosis (DVT), pulmonary embolism (PE), and unstable angina [1]. Major orthopedic surgery, including hip or knee replacement surgery or hip fracture surgery, are associated with a high risk of DVT/PE arterial and venous thrombosis. Therapy with a combination of various drugs is a necessary step in the treatment of arterial and venous thromboembolisms [3,4]. Direct and indirect anticoagulants are the basis for the prevention and treatment of venous thromboembolism (VTE) [5,6]. The former directly inhibit the serine proteases of the blood coagulation cascade, while the latter prevent the formation of precursors of prothrombin and other coagulation factors [7,8]. The general problem of the antithrombotic therapy is the difficulty of striking the optimal balance between efficacy and safety, with regard to bleeding

Methods

Results

Conclusion