Design and Synthesis of Novel Action Inhibitors for Bruton's Tyrosine Kinase (BTK)

布鲁顿酪氨酸激酶 (BTK) 新型作用抑制剂的设计与合成

• 批准号: 2751540
• 金额: --
• 依托单位: University of Cambridge
• 依托单位国家: 英国
• 项目类别: Studentship
• 财政年份: 2022
• 资助国家: 英国
• 起止时间: 2022 至 无数据
• 项目状态: 未结题
• 来源: https://gtr.ukri.org/projects?ref=studentship-2751540
• 关键词: Design; Synthesis; Novel; Action; Inhibitors

Bruton's Tyrosine Kinase (BTK) has a pivotal role in the BCR signalling pathway which is upregulated in B cell malignancies. Activation is dependent on two phosphorylation events, at Y551 and Y223 in the kinase and SH3 domains respectively. Of the remaining three domains, the Pleckstrin Homology (PH) domain is essential for the membrane recruitment which allows for phosphorylation. Conventional therapies target the ATP binding site in the kinase domain, specifically residue C481, to prevent BTK activation. However, this region is relatively conserved across the kinome and prolonged use leads to resistance and relapse as patients develop a C481S mutation. To overcome these issues, this project aims to exploit fragment-based drug discovery (FBDD) against the PH domain. Literature precedent justifies targeting this domain, which proved effective for a similar protein (AKT), as well as utilising FBDD against proteins with a significant role in cancer (CK2). Fragment elaboration led to a parent compound and the project is now well-established with three generations of compounds already synthesised by linking two hit fragments. Further generations of analogues will be synthesised and tested, using a synthetic route in need of optimisation. The scaffold will be investigated by replacing the ketone and phenyl groups and measuring affinity. Biochemical validation will be achieved in collaboration with the Hyvönen Group through mass spectrometry, differential scanning fluorimetry (DSF), X-ray crystallography and fluorescence polarisation (FP). Since the compounds are enantiomeric, key analogues will be sent to Astra Zeneca for separation. Testing the enantiomers individually in conjunction with molecular modelling and covalent docking will help deduce whether their binding affinities are competitive. Biological approaches to confirm inhibition will be investigated given the available facilities, but could include GFP-tagging, immunoprecipitation and blotting or flow cytometry. The research will contribute significantly to the field of covalent inhibition whilst providing a novel solution to the problem of resistance in targeting BTK.

Bruton的酪氨酸激酶（BTK）在BCR信号通路中具有关键作用，该途径在B细胞恶性肿瘤中进行了更新。激活分别取决于两个磷酸化事件，分别在激酶和SH3结构域中的Y551和Y223。在其余三个结构域中，Pleckstrin同源（pH）结构域对于允许磷酸化的膜募集至关重要。常规疗法靶向激酶结构域中的ATP结合位点合に行します。英语：但是，随着患者发展C481S突变，该区域相对在整个Kinome中配置相对构型，并且长时间使用会导致耐药性和继电器。为了克服这些问题，该项目旨在探索针对pH领域的基于碎片的药物发现（FBDD）。文献对该结构域的辩护是对类似蛋白质（AKT）有效的有效性，并利用FBDD对蛋白质在癌症中具有重要作用的蛋白质（CK2）（CK2）。片段阐述导致了父院，并且该项目现在已经建立了良好的，三代化合物已经通过链接两个命中片段来合成。使用需要优化的合成途径，将对其他几代类似物进行合成和测试。脚手架将通过取代酮和苯基并测量亲和力来研究脚手架。通过质谱，差异扫描荧光法（DSF），X射线晶体学和荧光极化（FP），将与Hyvönen组合作实现生化验证。由于这些化合物是对映体的，因此将键类似物发送到Astra Zeneca进行分离。与分子建模和共价对接分别测试对映异构体将有助于推断其结合亲和力是否具有竞争力。鉴于可用的设施，将研究确认抑制作用的生物学方法，但可能包括toging gfp，免疫沉淀和印迹或流式细胞仪。这项研究将极大地促进共价抑制领域，同时为靶向BTK的抗药性问题提供新颖的解决方案。