Tailoring Structures of Sulfated Oligosaccharides for Modulating Heparanase Activity

项目总结 乙酰肝素酶被认为是癌症侵袭性表型的主要调节因子， 是导致癌症患者预后不佳的重要因素，也是治疗的主要目标。 虽然基于碳水化合物的肝素酶已经被开发出来，但没有一种被翻译成 在临床上使用。此外，乙酰肝素酶与硫酸乙酰肝素(HS)的相互作用受 底物硫化序列，并且仅具有特定硫化模式的底物被切割 通过乙酰肝素酶。虽然最近已经解析了人乙酰肝素酶的晶体结构， 它仍然不允许确定具有定义的N-和O-的结合表位的结构 乙酰肝素酶各亚基的硫酸盐化模式。为了应对这些挑战，目标1需要 HS文库并行组合综合的新模块化学方法 三糖底物，代表所有可能的N-乙酰以及O-和N-硫化基序。 我们的策略提供了对乙酰肝素酶底物专一性的系统了解 乙酰肝素酶如何选择有利的切割位点。在目标2中，我们建议开发乙酰肝素酶- 抑制硫代低聚糖的高效和临床适用性。在《目标3》中，最 在AIM 2中开发的有效抑制剂将被测试与其他HS结合的交叉生物活性 负责调节抗凝剂和血管生成活性的蛋白质，抗体- 诱导的血小板减少和肿瘤细胞转移。最终目标是了解如果 乙酰肝素酶的体外抑制作用将在体内转化，我们将评估其疗效 抑制淋巴瘤肿瘤生长和实验性转移最有效的抑制剂(S)。 总而言之，这个项目将提供对硫酸盐识别主题的洞察和有利的 用于开发乙酰肝素酶抑制剂的切割位点。这个项目的重大影响是 如果成功，这可能会导致发现一种抑制肝素酶的小碳水化合物 用于治疗淋巴瘤的分子，淋巴瘤是北美第五大癌症， 主要发生在淋巴组织中的肿瘤。

PROJECT SUMMARY Heparanase is recognized as a master regulator of the aggressive phenotype of cancer, an important contributor to the poor outcome of cancer patients and a prime target for therapy. Although carbohydrate-based heparanase have been developed, but none were translated into use in the clinic. Additionally, interaction of heparanase with heparan sulfate (HS) is regulated by substrate sulfation sequences, and only substrates with specific sulfation patterns are cleaved by heparanase. Although the crystal structure of human heparanase has been recently resolved, it still does not allow determination of the structure of binding epitopes with defined N-and O- sulfation patterns at each subsite of heparanase. To address these challenges, Aim 1 entails a new modular chemical approach for the parallel combinatorial synthesis of a library of HS trisaccharide substrates, representing all possible N-acetyl as well as O- and N-sulfation motifs. Our strategy provides a systematic understanding of substrate specificity for heparanase and how heparanase selects favorable cleavage site. In Aim 2, we propose to develop heparanase- inhibiting sulfated oligosaccharides of high efficacy and clinical applicability. In Aim 3, the most potent inhibitors developed in Aim 2 will be tested for cross-bioactivity to other HS-binding proteins, which are responsible for mediating anticoagulant and angiogenic activity, antibody- induced thrombocytopenia, and tumor cell metastasis. With the ultimate goal of understanding if the in vitro inhibition of heparanase would translate in vivo, we will evaluate the effectiveness of the most potent inhibitor(s) in inhibiting lymphoma tumor growth and experimental metastasis. Together, this project will provide insight into sulfate-recognition motifs and favorable cleavage site for use to develop inhibitors of heparanase. The significant impact of this project is that, if successful, it could lead to the discovery of a heparanase-inhibiting small carbohydrate molecule for treatment of lymphomas, which are the fifth leading cancer in the North America, produce tumors predominantly in lymphoid structures.