Cellular and biophysical mechanisms of small-molecule inhibition of non-structural protein 5A.

非结构蛋白 5A 小分子抑制的细胞和生物物理机制。

• 批准号: 2398038
• 金额: --
• 依托单位: University College London
• 依托单位国家: 英国
• 项目类别: Studentship
• 财政年份: 2020
• 资助国家: 英国
• 起止时间: 2020 至 无数据
• 项目状态: 未结题
• 来源: https://gtr.ukri.org/projects?ref=studentship-2398038
• 关键词: Cellular; biophysical; mechanisms; small; molecule

Intrinsically disordered proteins (IDPs) play important roles in diseases such as Alzheimer's, breast cancer, and viruses, including Hepatitis C virus (HCV) and SARS-CoV-2. As such, targeting disorder offers huge therapeutic opportunity. However, IDPs are often considered undruggable due to their highly dynamic nature and the absence of well-defined structure. Non-structural protein 5A (NS5A) is a multi-functional phosphoprotein from HCV, a virus which affects over 58 million people worldwide and causes chronic liver disease. Domain I of NS5A is highly structured and exclusively involved in genome replication, while domains II and III are unstructured and required for the production of infectious viral particles. Existing therapies in widespread clinical use comprise of direct acting antivirals (DAAs) which are believed to target NS5A. These drugs are exceptionally potent, exhibiting picomolar activities, however their exact mechanisms of action remain unknown. Preliminary nuclear magnetic resonance (NMR) experiments undertaken in Professor Hansen's lab show that one such drug, ledipasvir, interacts with the unstructured region of NS5A.This PhD project aims to uncover the cellular and biophysical mechanisms of NS5A inhibition. I aim to understand whether existing therapeutics target the structured and/or unstructured domains of NS5A, and if these interactions result in inhibition of HCV replication. In the Towers lab, an HCV replicon assay will be employed to test ledipasvir and other DAAs against HCV replication and determine the significance of the disordered and structured regions of NS5A on the activity of the drugs. NMR experiments, undertaken in the Hansen lab, will be used to characterise the binding of these drugs to various domains of NS5A. This research will deepen understanding of the therapeutic activity of NS5A-targeting DAAs and be important both in the treatment of HCV, and in the design of drugs targeting other proteins, particularly those with disordered regions.

内含子无序蛋白（IDP）在阿尔茨海默病、乳腺癌等疾病和病毒（包括丙型肝炎病毒（HCV）和SARS-CoV-2）中发挥重要作用。因此，靶向疾病提供了巨大的治疗机会。然而，由于国内流离失所者的高度动态性和缺乏明确界定的结构，他们往往被认为是不可求助的。非结构蛋白5A（NS 5A）是来自HCV的多功能磷蛋白，HCV是一种影响全球超过5800万人并导致慢性肝病的病毒。NS 5A的结构域I是高度结构化的并且专门参与基因组复制，而结构域II和III是非结构化的并且是产生感染性病毒颗粒所需的。广泛临床使用的现有疗法包括据信靶向NS 5A的直接作用抗病毒药物（DAA）。这些药物非常有效，表现出皮摩尔活性，但其确切的作用机制仍然未知。在汉森教授实验室进行的初步核磁共振（NMR）实验表明，一种这样的药物ledipasvir与NS 5A的非结构化区域相互作用。我的目的是了解现有的治疗方法是否靶向NS 5A的结构化和/或非结构化结构域，以及这些相互作用是否导致HCV复制的抑制。在Towers实验室，将采用HCV复制子检测来测试ledipasvir和其他DAA对HCV复制的影响，并确定NS 5A的无序和结构化区域对药物活性的重要性。在汉森实验室进行的核磁共振实验将被用来验证这些药物与NS 5A不同结构域的结合。这项研究将加深对NS 5A靶向DAA的治疗活性的理解，并且在治疗HCV和设计靶向其他蛋白质，特别是那些具有无序区域的蛋白质的药物方面都很重要。