Luminescent Host Molecules for Multisite Recognition of Polyphosphate Anions

新药的研究通常从先导化合物的高通量筛选开始，然后确定潜在药物的作用方式，并测量选择性和效力。许多药物通过抑制酶活性起作用，因此，为了增加对先导药物化合物选择的信心，制药公司必须有可靠的、负担得起的测定方法来准确测量酶活性。消耗和产生多磷酸核苷阴离子的酶是癌症药物发现的主要目标。这将使最佳候选药物能够在进入昂贵的动物和人体试验之前被确定。大多数现有的酶分析需要昂贵的，不稳定的抗体或化学修饰的试剂，这是耗时的准备和需要特别小心处理。这些试剂的高成本和验证测定所需的时间给药物开发过程带来了巨大的压力。此外，这些检测仅限于单一“终点”测量，这限制了我们对新药作用方式的理解。这增加了候选药物通过早期开发阶段的风险，只是在后期，更昂贵的阶段失败。为了在药物发现过程的早期提高生产力，迫切需要一种低成本的酶活性实时监测方法。这个项目的目的是开发分子探针，可以可逆地结合特定的核苷多磷酸阴离子（如二磷酸腺苷），这些阴离子是在药物上重要的酶反应中产生的。在与阴离子结合后，探针将提供一个发光信号，实时精确地指示酶的活性。该探针将用于直接测量几种重要酶类（激酶、gtp酶、糖基转移酶）常见的多磷酸阴离子的产生，从而消除了对昂贵的抗体或化学修饰试剂的需要。所提出的分子探针在降低进行高通量筛选分析所需的成本和时间方面具有巨大的潜力。它们将为快速、准确地测定酶的动力学和机制提供重要的一步。这将有助于在药物发现的早期阶段更好地选择和验证新的候选药物，减少在更漫长和昂贵的动物和人体试验阶段寻找注定失败的化合物的努力。

The search for new drugs often begins with high-throughput screening of lead compounds followed by determination of mode of action of the potential drug and measurements of selectivity and potency. Many drugs act by inhibiting enzyme activity, therefore, to increase confidence in the selection of lead drug compounds it is crucial that pharmaceutical companies have robust, affordable assays to measure enzyme activity accurately. Enzymes that consume and produce nucleoside polyphosphate anions represent a major target in cancer drug discovery. This will enable the best drug candidates to be identified before they enter expensive animal and human testing.The majority of existing enzyme assays require expensive, unstable antibodies or chemically modified reagents, which are time consuming to prepare and require special care in handling. The high cost of these reagents and time required to validate the assays places a significant strain on the drug development process. In addition, these assays are restricted to single 'end-point' measurements, which limits our understanding of the mode of action of a new drug. This increases the risk of a drug candidate passing the early development stages, only to fail at a later, more expensive stage. In order to increase productivity earlier in the drug discovery process, a low-cost method for real-time monitoring of enzyme activity is urgently needed.The aim of this project is to develop molecular probes that bind reversibly to specific nucleoside polyphosphate anions (e.g. adenosine diphosphate) that are produced during pharmaceutically important enzyme reactions. Upon binding to the anion, the probe will provide a luminescent signal that precisely indicates the activity of the enzyme in real-time. The probes will be used to directly measure the production of polyphosphate anions common to several important enzyme classes (kinases, GTPases, glycosyltransferases), eliminating the need for expensive antibodies or chemically modified reagents. The proposed molecular probes have enormous potential to reduce the cost and time required to conduct high-throughput screening assays. They will provide a vital step towards the rapid, accurate determination of enzyme kinetics and mechanism. This will enable better selection and validation of new drug candidates at an early stage in drug discovery, reducing effort pursuing compounds destined to fail in the more lengthy and costly phases of animal and human testing.