Identification and elaboration of fluorine containing inhibitors of proline rich tyrosine kinase and endothelial nitric oxide synthase

2型糖尿病是一种越来越常见的疾病，我们的身体不能产生足够的胰岛素，或者产生的胰岛素功能不正常。英国约有400万人受到2型糖尿病的影响，据估计，仅在英国，每年用于治疗糖尿病及其并发症的费用就高达140亿英镑。与没有糖尿病的人相比，糖尿病患者患心血管疾病的可能性高达五倍，他们的生活质量可能受到更大的影响。这在一定程度上是因为糖尿病改变了血管内的细胞，导致血管中更多的有毒分子，以及一氧化氮等有益分子的减少。我们最近发现，在2型糖尿病患者体内发现了一种名为PYK2的蛋白质水平升高。这种蛋白质通过阻止产生一氧化氮的蛋白质(称为eNOS)工作来降低一氧化氮水平。我们将使用基于片段的配体发现来识别调节eNOS和PYK2之间相互作用的片段。这些可以被细化成小分子。与传统的筛选技术相比，基于片段的配体发现允许对化学空间进行更大的采样，即使使用中等大小的化合物文库也是如此。由于片段通常表现出较弱的结合亲和力，因此需要使用核磁共振等敏感的生物物理技术来检测结合事件。与不含氟的片段相比，使用含氟片段进行核磁共振筛选可提高吞吐量，因为获得的光谱更简单且更易于阅读。这使我们能够筛选没有光谱重叠的样品混合物，允许表征碎片的鸡尾酒，每管约10种化合物，最大限度地提高效率。这种新发现的相互作用为解决2型糖尿病的心血管并发症提供了一种新的方法。利兹大学的新型4通道核磁共振探头首次实现了含氟片段的筛查。这项工作的结果将为开发针对以前未开发的途径治疗2型糖尿病的疗法铺平道路。

Type 2 Diabetes Mellitus, where our bodies do not produce enough insulin, or the insulin that is produced does not function properly,is an increasingly common disorder. Around four million people in the UK are affected by type 2 diabetes and it is estimated that 14 billion pounds is spent a year on treating diabetes and its complications in the UK alone. People with diabetes are up to five times more likely to have cardiovascular disease and their quality of life is likely to be more affected by it than people who don't have the disease.This is in part due to diabetes changing the cells that line your blood vessels, leading more toxic molecules in them and a decrease in beneficial molecules such as nitric oxide. We have recently shown that increased levels of a protein called PYK2 is found in the bodies of people with type 2 diabetes. This protein reduces nitric oxide levels by preventing the protein that makes nitric oxide (called eNOS) from working. We will use fragment based ligand discovery to identify fragments which modulate the interaction between eNOS and PYK2. These can be elaborated into small molecules. Fragment based ligand discovery allows greater sampling of chemical space compared with traditional screening techniques, even with a modestly sized compound library. Because of the weaker binding affinities typically exhibited by fragments, sensitive biophysical techniques such as Nuclear Magnetic Resonance are required to detect the binding event. The use of fluorine containing fragments for Nuclear Magnetic Resonance screening allows increased throughput versus fragments without a fluorine in as the spectra obtained are simplified and easier to read. This gives us the ability to screen mixtures of samples without spectral overlap, allowing cocktails of fragments, with around 10 compounds per tube, to be characterised, maximising efficiency.This newly discovered interaction offers a novel way to address the cardiovascular complications of type 2 diabetes. The new 4 channel Nuclear Magnetic Resonance probe at Leeds allows fluorine containing fragment screening for the first time. Results from this work will pave the way to develop therapies to treat type 2 diabetes targeting a previously unexploited pathway.