Organosulfates and Polyphosphates: Their Synthesis and the Enzymes that Process Them.

我的研究计划的主要重点是开发合成分子的新方法，这些分子可用于构建生物学上重要的化合物，并评估选定的化合物作为治疗剂设计的先导结构，并阐明具有医学意义的酶的机制。 本提案的第一个主题涉及硫酸化碳水化合物的合成。 这是一类非常重要的生物分子，因为它们在许多重要的生物过程中发挥关键作用，如血液凝固和免疫反应。 我们已经开发了一种新的方法来制备这些化合物，采用单糖轴承保护磺基二酯作为积木。为了确定该方法的范围，将进行关于受保护的磺基二酯区域选择性掺入单糖结构单元以及受保护的磺基二酯对糖基化反应的影响的研究。 两个复杂的多硫酸化寡糖的全合成也将实现。 该提案的第二部分涉及类固醇硫酸酯酶（STS）抑制剂的设计，合成和评价，STS是一种参与雌激素储存的酶。STS的抑制剂正被研究作为治疗乳腺癌的治疗剂。我们已经制备了非常有效的STS可逆和不可逆抑制剂。我们将确定这些抑制剂如何结合STS使用光亲和标记，建模，NMR和X射线晶体学分析。本论文亦将探讨STS中的底物进入及产物释放机制。 在本提案的第三部分，中间体和过渡态类似物的反应催化的胞苷三磷酸合酶（CTPS），一种抗癌和抗病毒的目标，将构建使用一种新的和强大的磷酸化程序最近在我们的小组。 这些化合物将用于确定哪些残基对于CTPS反应中的配体结合以及中间体和过渡态稳定是重要的。这些研究的结果将为开发靶向CTPS的药物提供至关重要的信息。

The primary focus of my research program is to develop new methods for synthesizing molecules that can be used in the construction of biologically important compounds and evaluate selected compounds as lead structures for the design of therapeutic agents and for elucidating mechanisms of medicinally significant enzymes. The first subject of this proposal concerns the synthesis of sulfated carbohydrates. This is an extremely important class of biomolecules as they play key roles in numerous important biological processes such as blood clotting and the immune response. We have developed a new approach to preparing these compounds which employs monosaccharides bearing protected sulfodiesters as building blocks. To determine the scope of this methodology, studies on the regioselective incorporation of protected sulfodiesters into monosaccharide building blocks as well as the effect of protected sulfodiesters on glycosylation reactions will be performed. The total synthesis of two complex multisulfated oligosaccharides will also be achieved. The second part of this proposal deals with the design, synthesis and evaluation of inhibitors of steroid sulfatase (STS), an enzyme involved in the storage of estrogens. Inhibitors of STS are being examined as therapeutics for treating of breast cancer. We have prepared very potent reversible and irreversible inhibitors of STS. We will determine how these inhibitors bind to STS using photoaffinity labeling, modeling, NMR and x-ray crystallographic analyses. The mechanism of substrate entry and product release in STS will also be studied. In the third part of this proposal intermediate and transition state analogs of the reaction catalyzed by cytidine triphosphate synthase (CTPS), an anticancer and antiviral target, will be constructed using a novel and powerful phosphorylation procedure recently developed in our group. These compounds will be used to determine which residues are important for ligand binding and intermediate and transition state stabilization in the CTPS reaction. These results of these studies will provide information that will be crucial for developing drugs that target CTPS.