Expanding the role of P-31 NMR in the study of phosphate-acting enzymes

布鲁克是世界领先的核磁共振（NMR）光谱仪，并热衷于扩大其在催化和酶抑制研究中的存在。许多药物靶标是化学转化含磷酸盐化合物的酶。12种激酶抑制剂药物已经用于癌症化疗，还有100多种正在临床试验中。对酶机制的详细了解极大地帮助了新的抑制剂的设计：流感药物奥司他韦和扎那米韦，以及抗癌药物福罗地辛都是在确定靶酶的过渡态（TS）结构后设计的。TS是酶必须跨越的能量屏障的顶端，酶才能进行反应。酶与TSs结合非常紧密，**使TS模仿有效的抑制剂。然而，确定TS结构在技术上要求很高；它要求**测量同位素标签对反应速率的影响，这种影响通常小于1%。核磁共振波谱学的最新进展使大多数实验室都可以使用这些测量仪器，但是利用这些新功能所需的方法仍然不发达。我们建议结合物理化学/应用酶学研究，开发利用磷-31 （31P）核磁共振测量**同位素效应的方法。这代表了一个合适的专业知识组合，布鲁克加拿大提供**NMR专业知识，而Berti实验室提供TS分析专业知识。Bruker Canada将直接受益于新方法的开发，这些方法将成为其应用程序库的一部分。私营部门和学术实验室都将能够利用核磁共振技术的扩展曲目来确定**TS结构，扩大非专业实验室的核磁共振光谱的效用。布鲁克加拿大公司还将直接获得麦克马斯特的样品制备、HQP和核磁共振光谱仪，为潜在客户进行开发和演示。所提出的核磁共振实验有望吸引其他从事抑制剂设计和反应机制研究的学术实验室的兴趣。因此，拟议中的工作将扩大布鲁克加拿大公司在私营和公共部门的客户群。

Bruker is the world leader in nuclear magnetic resonance (NMR) spectrometers, and is keen to expand its**presence in catalysis and enzyme inhibition research. Many pharmaceutical targets are enzymes that**chemically transform phosphate-containing compounds. Twelve kinase inhibitor drugs are already used in**cancer chemotherapy, and >100 more in are clinical trials. Detailed understanding of enzyme mechanisms**greatly aids new inhibitor design: the flu drugs oseltamivir and zanamivir, and the cancer drug forodesine were**all designed after determining the target enzymes' transition state (TS) structures. The TS is the top of the**energetic barrier that enzymes must surmount for a reaction to occur. Enzymes bind TSs extremely tightly,**making TS mimics potent inhibitors. However, determining TS structures is technically demanding; it requires**measuring the effects of isotopic labels on reaction rates, effects that are frequently < 1%. Recent advances in**NMR spectroscopy have brought the instrumentation for these measurements within reach of most labs, but the**methods needed to exploit these new capabilities remain under-developed. We propose a combined physical**chemistry / applied enzymology study to develop methods to use phosphorus-31 (31P) NMR to measure**isotope effects. This represents an opportune combination of expertise, with Bruker Canada providing the**NMR expertise while the Berti lab providing expertise in TS analysis. Bruker Canada will directly benefit**from the development of new methods that will become part of their applications library. Both private sector**and academic labs will be able to take advantage of the expanded repertoire of NMR techniques to determine**TS structures, expanding the utility of NMR spectroscopy for non-specialist labs. Bruker Canada will also**benefit from direct access to sample preparation, HQP and the NMR spectrometers available at McMaster for**both development and demonstrations for potential customers. The proposed NMR experiments are expected**to attract the interest of other academic laboratories working on inhibitor design and reaction mechanisms. The**proposed work will therefore expand Bruker Canada's customer base in both the private and public sectors.