Structural features of glycol-split low-molecular-weight heparins and their heparin lyase generated fragments.
Structural features of glycol-split low-molecular-weight heparins and their heparin lyase generated fragments.
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DOI：
10.1007/s00216-013-7446-4
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发表时间：
2014-01
影响因子：
4.3
通讯作者：
Naggi, Annamaria
中科院分区：
文献类型：
--
作者：
Alekseeva, Anna;Casu, Benito;Cassinelli, Giuseppe;Guerrini, Marco;Torri, Giangiacomo;Naggi, Annamaria
关键词：
高碘酸盐氧化，然后硼氢化物还原，将众所周知的抗血栓形成剂肝素和低分子量肝素（LMWH）转化为它们的“还原的氧肝素”（RO）类型的“二醇裂解”（gs）衍生物，其中一些目前正在开发为潜在的抗癌和抗炎药物。尽管GS-肝素的结构最近已被研究，但更复杂和生物利用度更高的GS-LMWH的细节尚未报道。我们获得了RO衍生物的三个最常见的低分子量肝素（汀扎肝素，依诺肝素，达肝素），并研究了它们的结构，通过二维核磁共振（2D NMR）光谱和离子对反相高效液相色谱（IPRP-HPLC）与电喷雾电离质谱（ESI-MS）。将LC-MS分析扩展至其肝素酶生成的寡糖。RO-LMWH的联合NMR/LC-MS分析提供了主要涉及内部非硫酸化葡萄糖醛酸和艾杜糖醛酸残基（包括部分水解形成“残留物”）的二醇裂解诱导转化的证据，以及在非还原末端（主要在RO-达肝素中）形成的葡萄糖醛酸残基水解的证据。还获得了微小修饰的证据，如一些达肝素内部氨基糖残基的环收缩。出乎意料的是，发现依诺肝素还原端的N-硫酸化1，6-脱水甘露糖胺残基对高碘酸盐氧化敏感。此外，在亭扎肝素和依诺肝素中，硼氢化物还原将还原端的半缩醛氨基糖转化为糖醇。在用肝素酶消化之前和之后，单个LMWH的RO衍生物的典型LC-MS特征包括从原始抗凝血酶结合和“连接”区域产生的寡糖。
Periodate oxidation followed by borohydride reduction converts the well-known antithrombotics heparin and low-molecular weight heparins (LMWHs) into their “glycol-split” (gs) derivatives of the “reduced oxyheparin” (RO) type, some of which are currently being developed as potential anti-cancer and anti-inflammatory drugs. Whereas the structure of gs-heparins has been recently studied, details of the more complex and more bioavailable gs-LMWHs have not been yet reported. We obtained RO derivatives of the three most common LMWHs (tinzaparin, enoxaparin, and dalteparin) and studied their structures by two-dimensional nuclear magnetic resonance (2D NMR) spectroscopy and ion-pair reversed-phase high-performance liquid chromatography (IPRP-HPLC) coupled with electrospray ionization mass spectrometry (ESI-MS). The LC-MS analysis was extended to their heparinase-generated oligosaccharides. The combined NMR/LC-MS analysis of RO-LMWHs provided evidence for glycol-splitting-induced transformations mainly involving internal nonsulfated glucuronic and iduronic acid residues (including partial hydrolysis with formation of “remnants”), and for the hydrolysis of the gs uronic acid residues when formed at the non-reducing ends (mainly, in RO-dalteparin). Evidence for minor modifications, such as ring contraction of some dalteparin internal aminosugar residues was also obtained. Unexpectedly, the N-sulfated 1,6-anhydro-mannosamine residues at the enoxaparin reducing end were found to be susceptible to the periodate oxidation. In addition, in tinzaparin and enoxaparin the borohydride reduction converts the hemiacetalic aminosugars at the reducing end to alditols. Typical LC-MS signatures of RO-derivatives of individual LMWH both before and after digestion with heparinases included oligosaccharides generated from the original antithrombin-binding and “linkage” regions.
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影响因子：
6.9
作者：
Casu, Benito;Naggi, Annamaria;Torri, Giangiacomo
通讯作者：
Torri, Giangiacomo
影响因子：
6.7
作者：
Bisio, Antonella;Vecchietti, Davide;Torri, Giangiacomo
通讯作者：
Torri, Giangiacomo
影响因子：
5.7
作者：
Guerrini, Marco;Guglieri, Sara;Torri, Giangiacomo
通讯作者：
Torri, Giangiacomo
影响因子：
6.7
作者：
Mousa, Shaker A.;Linhardt, Robert;Amirkhosravi, Ali
通讯作者：
Amirkhosravi, Ali
影响因子：
3.5
作者：
HOOK, M;BJORK, I;LINDAHL, U
通讯作者：
LINDAHL, U