Preparation and evaluation a new generation of low molecular weight heparin

Abstract

Enoxaparin is widely used in clinic, but it has some disadvantages. For example, its anticoagulant activity is weaker compared with heparin and it can not be effectively neutralizad by protamine sulfate (PS) in case of bleeding. Therefore, in this work, a new generation of low molecular weight heparin (NG-LMWH) was prepared.The NG-LMWH was prepared with the method of alkaline β-elimination followed by gel chromatography. Estimating the molecular weight of the NG-LMWH by GPC-HPLC, it has a remarkably low polydispersity index and narrow molecular weight distribution. The polydispersity index of NG-LMWH was 1.052, which was lower than heparin (1.5) and enoxaparin (1.279). Anti-FXa and anti-FIIa potency of NG-LMWH was much higher than that of Enoxaparin, and close to that of heparin, which was determined by chromogenic substrate method. To test the degree of anti-FXa or anti-FIIa potency neutralized by PS, equivalent anti-FXa or anti-FIIa activity doses of different anticoagulant in plasma were titrated with increasing amounts of PS in plasma. The results indicate that NG-LMWH was more efficiently neutralized by PS than enoxaparin.The efficacy of anti-thrombus of NG-LMWH was superior to enoxaparin and the effect was dose dependent, which was evaluated with rat carotid artery thrombosis and inferior vena cava thrombosis model. The results of pharmacokinetics in New Zealand rabbits showed that the pharmacokinetic characteristics of NG-LMWH were similar to enoxaparin. The NG-LMWH prepared in this work has both advantages of heparin and enoxaparin with more effective and safer anticoagulation than enoxaparin.

Keywords: Anti-thrombus; Anticoagulation; Low molecular weight heparin (LMWH); Pharmacokinetic; Protamine sulfate; Rational design.