Key Laboratory of Marine Drugs, Chinese Ministry of Education, Institute of Marine Drug and Food, Ocean University of China, Qingdao, 266003, China.
J Med Chem. 2011 Jan 27;54(2):603-10. doi: 10.1021/jm101381k. Epub 2010 Dec 17.
Seven pharmaceutical heparins were investigated by oligosaccharide mapping by digestion with heparin lyase 1, 2, or 3, followed by high performance liquid chromatography analysis. The structure of one of the prepared mapping standards, ΔUA-Gal-Gal-Xyl-O-CH(2)CONHCH(2)COOH (where ΔUA is 4-deoxy-α-l-threo-hex-4-eno-pyranosyluronic acid, Gal is β-d-galactpyranose, and Xyl is β-d-xylopyranose) released from the linkage region using either heparin lyase 2 or heparin lyase 3 digestion, is reported for the first time. A size-dependent susceptibility of site cleaved by heparin lyase 3 was also observed. Heparin lyase 3 acts on the undersulfated domains of the heparin chain and does not cleave the linkages within heparin's antithrombin III binding site. Thus, a novel low molecular weight heparin (LMWH) is afforded on heparin lyase 3 digestion of heparin due to this unique substrate specificity, which has anticoagulant activity comparable to that of currently available LMWH.
七种药物肝素通过肝素酶 1、2 或 3 的消化进行寡糖图谱分析，然后进行高效液相色谱分析。在所制备的图谱标准品之一的结构中，用肝素酶 2 或肝素酶 3 消化从连接区释放出 ΔUA-Gal-Gal-Xyl-O-CH(2)CONHCH(2)COOH（其中 ΔUA 是 4-脱氧-α-l-赤式-己-4-烯吡喃糖醛酸，Gal 是β-d-半乳糖吡喃糖，Xyl 是β-d-木吡喃糖），这是首次报道。还观察到肝素酶 3 切割位点的尺寸依赖性易感性。肝素酶 3 作用于肝素链的低硫酸化结构域，并且不会切割抗凝血酶 III 结合部位内的肝素键。因此，由于这种独特的底物特异性，肝素酶 3 消化肝素可提供新型低分子量肝素（LMWH），其抗凝活性可与目前可用的 LMWH 相媲美。
