Hirudin biosimilar(Rhein-Minapharm)

Current antithrombotic therapies face dual constraints of bleeding complications and monitoring requirements. Although natural hirudin provides targeted thrombin inhibition, its clinical adoption is hindered by sourcing limitations. This study developed a recombinant hirudin variant HMg (rHMg) with enhanced anticoagulant activity through genetic engineering and established cost-effective large-scale production methods. The synthesised HMg gene was expressed in E. coli BL21 via a pET vector plasmid, followed by nickel-affinity purification. Systematic evaluations demonstrated rHMg's antithrombin activity of 9573 ATU/mg, dose-dependent prolongation of APTT/PT/TT. It has superior thrombin inhibition with the IC₅₀ and K_i values were 2.8 and 0.323 nM respectively compared to FDA approved drug bivalirudin (p < 0.001). The high-yield prokaryotic expression of rHMg with enhanced anticoagulant efficacy provides a novel strategy for developing affordable antithrombotic drugs, showing significant potential for cardiovascular disease management.

Renal fibrosis is a pathological feature of chronic kidney injury that contributes to renal failure. This study aimed to explore the effects of Hirudin on renal fibrosis. The antifibrotic effect of Hirudin was evaluated using unilateral ureteral obstruction (UUO) rats and TGF-β-treated HK-2 cells. The autophagy inhibitor 3-methyladenine was used to further explore the potential mechanism. Hirudin treatment significantly reduced UUO-induced elevations in blood urea nitrogen, creatinine, and alpha-smooth muscle actin (α-SMA) levels and improved kidney injury and renal fibrosis. In addition, Hirudin markedly decreased NLRP3 inflammasome-related protein expression and increased autophagy-related protein expression in the kidneys of UUO rats. Hirudin significantly increased cell viability, reduced α-SMA and NLRP3 inflammasome-related protein levels, and increased autophagy-related protein levels in TGF-β-treated HK-2 cells. However, the effects of Hirudin were counteracted by 3-methyladenine. In conclusion, Hirudin inhibits the activation of NLRP3 inflammasome by inducing autophagy to improve renal fibrosis.