Preactivation-based, one-pot combinatorial synthesis of heparin-like hexasaccharides for the analysis of heparin-protein interactions.
Preactivation-based, one-pot combinatorial synthesis of heparin-like hexasaccharides for the analysis of heparin-protein interactions.
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DOI：
10.1002/chem.201000987
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发表时间：
2010-07-26
影响因子：
4.3
通讯作者：
Huang, Xuefei
中科院分区：
文献类型：
--
作者：
Wang, Zhen;Xu, Yongmei;Yang, Bo;Tiruchinapally, Gopinath;Sun, Bin;Liu, Renpeng;Dulaney, Steven;Liu, Jian;Huang, Xuefei
肝素（HP）和硫酸乙酰肝素（HS）在许多生物事件中起重要作用。越来越多的证据表明，HP和HS的生物学功能可能严重依赖于它们的精确结构，包括艾杜糖醛酸的位置和硫酸化模式。然而，由于巨大的结构变化，解开HP代码一直极具挑战性。为了克服这一障碍，我们研究了使用基于预活化的一锅糖基化方法组装HP/HS寡糖库的可能性。HP/HS寡糖合成中的主要挑战是在葡糖胺和糖醛酸之间形成顺式-1，4-键的立体选择性。通过筛选，在匹配的糖基供体和受体上鉴定出合适的保护基团，导致HP中存在的顺式-1，4-和反式-1，4-键的立体特异性形成。所设计的保护基化学也非常灵活。从两个先进的硫代糖基二糖中间体，所有所需的二糖模块库制备可以产生在一个发散的方式，这大大简化了积木的制备。此外，基于预激活的一锅法的反应独立性使我们能够混合构建模块。这允许以组合方式快速组装具有系统变化和精确控制的主链结构的十二种HP/HS六糖。在糖组装中实现的速度和高产率，而不需要使用大量过量的构建块，突出了我们的方法的优点，这可以普遍用于促进HP/HS生物学的研究。作为原理的证明，该六糖组用于探测骨架序列对与成纤维细胞生长因子-2（FGF-2）结合的影响。经鉴定，侧翼为N-硫酸化葡糖胺的2-O-硫酸化艾杜糖醛酸的三糖序列是最小结合基序，发现N-硫酸化是关键。这为进一步开发对FGF-2结合更有效的化合物提供了有用的信息，这些化合物在伤口愈合和抗癌治疗中具有潜在的应用。
Heparin (HP) and heparan sulfate (HS) play important roles in many biological events. Increasing evidence has shown that the biological functions of HP and HS can be critically dependent upon their precise structures, including the position of the iduronic acids and sulfation patterns. However, unraveling the HP code has been extremely challenging due to the enormous structural variations. To overcome this hurdle, we investigated the possibility of assembling a library of HP/HS oligosaccharides using a preactivation-based, one-pot glycosylation method. A major challenge in HP/HS oligosaccharide synthesis is stereoselectivity in the formation of the cis-1,4-linkages between glucosamine and the uronic acid. Through screening, suitable protective groups were identified on the matching glycosyl donor and acceptor, leading to stereospecific formation of both the cis-1,4- and trans-1,4-linkages present in HP. The protective group chemistry designed was also very flexible. From two advanced thioglycosyl disaccharide intermediates, all of the required disaccharide modules for library preparation could be generated in a divergent manner, which greatly simplified building-block preparation. Furthermore, the reactivity-independent nature of the preactivation-based, one-pot approach enabled us to mix the building blocks. This allowed rapid assembly of twelve HP/HS hexasaccharides with systematically varied and precisely controlled backbone structures in a combinatorial fashion. The speed and the high yields achieved in glycoassembly without the need to use a large excess of building blocks highlighted the advantages of our approach, which can be of general use to facilitate the study of HP/HS biology. As a proof of principle, this panel of hexasaccharides was used to probe the effect of backbone sequence on binding with the fibroblast growth factor-2 (FGF-2). A trisaccharide sequence of 2-O-sulfated iduronic acid flanked by N-sulfated glucosamines was identified to be the minimum binding motif and N-sulfation was found to be critical. This provides useful information for further development of more potent compounds towards FGF-2 binding, which can have potential applications in wound healing and anticancer therapy.
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影响因子：
2.8
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