Hirudin variants production by genetic engineered microbial factory

Abstract

Hirudin was discovered as an active anticoagulant in leech extracts almost 60 years ago. Since their initial discovery, hirudin and its variants have been produced with various anti-thrombotic, cancer cell inhibition, diabetic cataract treatment and anti-fatigue activities. Some hirudin variants have been approved for clinical use and released into the marketplace. Recent progress has seen made in relation to hirudin variants expressed in several well-established microbial hosts, including Escherichia coli, Saccharomyces cerevisiae, Pichia pastoris and others, with high levels of activity and yield. This review summarizes the current progress on hirudin production using microbial producers, and considers the outlook for future development.

The efficacy and safety of Hirudin plus Aspirin versus Warfarin in the secondary prevention of Cardioembolic Stroke due to Nonvalvular Atrial Fibrillation: A multicenter prospective cohort study

“…The recombinant hirudin products, such as lepirudin, desirudin and bivalirudin, have manifested their clinical relevance in short term preoperative and postoperative thrombosis prophylaxis [21], disseminated intravascular coagulation [77], coronary ischemic syndromes [25], extracorporeal circulation [78], and heparin-induced thrombocytopenia [79]. Currently, bivalirudin, a synthetic hirudin analog comprising 20 amino acid residues from hirudin, shows increasing importance in clinical use for its lower immunogenicity and less dependence on renal clearance [80]. Bivalirudin has been approved in the use of percutaneous transluminal coronary angioplasty [81].…”

Cell-free synthesis of the hirudin variant 1 of the blood-sucking leech Hirudo medicinalis

“…The large-scale synthesis and purification of hirudin for scientific, commercial and medical purposes remains a challenging task 16 , 23 . Preparations of hirudin from whole leeches may lead to ethical concerns (death of innumerable animals) and/or limitations due to the collapse of natural populations 2 , 53 .…”

“…Preparations of hirudin from whole leeches may lead to ethical concerns (death of innumerable animals) and/or limitations due to the collapse of natural populations 2 , 53 . The biotechnological production of hirudin in bacteria, yeast and animal cells is hence an attractive alternative 16 . However, there are major drawbacks and limitations in heterologous expression, too 54 .…”

“…Hirudin is a drug of medical relevance in clinical use for decades 67 , 68 . So far, the biotechnological production of recombinant hirudin depends on either bacterial or yeast expression systems 16 . Both systems have major drawbacks in terms of putative contaminations and limitations in terms of yield of biologically active product 34 , 35 , 37 .…”

“…The availability of mRNA/cDNA sequence data of hirudin completely altered the situation and enabled the production of large amounts of recombinant hirudin in prokaryotic and eukaryotic expression systems. Bacterial expression systems include Escherichia coli ( E. coli ) 7 – 10 , Bacillus subtilis , Lactococcus lactis and Streptomyces lividans 11 , 12 , whereas in fungi hirudin could be successfully synthesized in Acremonium chrysogenum , Ogataea angusta , Pichia pastoris and Saccharomyces cerevisiae 13 – 16 , respectively. Eukaryotic expression systems, beside fungi, include insect cell lines, Chinese hamster ovary (CHO) cells, mouse fibroblast cells, porcine endothelial cells and pituitary secretory cell lines 17 – 19 .…”