a Centro de Investigaciones Biológicas (CIB-CSIC), C/Ramiro de Maeztu , Madrid , Spain.
b Department of Biochemistry , University of Cambridge , Cambridge , UK.
J Enzyme Inhib Med Chem. 2018 Dec;33(1):1034-1047. doi: 10.1080/14756366.2018.1476502.
Allosteric sites on proteins are targeted for designing more selective inhibitors of enzyme activity and to discover new functions. Acetylcholinesterase (AChE), which is most widely known for the hydrolysis of the neurotransmitter acetylcholine, has a peripheral allosteric subsite responsible for amyloidosis in Alzheimer's disease through interaction with amyloid β-peptide. However, AChE plays other non-hydrolytic functions. Here, we identify and characterise using computational tools two new allosteric sites in AChE, which have allowed us to identify allosteric inhibitors by virtual screening guided by structure-based and fragment hotspot strategies. The identified compounds were also screened for in vitro inhibition of AChE and three were observed to be active. Further experimental (kinetic) and computational (molecular dynamics) studies have been performed to verify the allosteric activity. These new compounds may be valuable pharmacological tools in the study of non-cholinergic functions of AChE.
蛋白质的别构位点是设计酶活性更具选择性抑制剂和发现新功能的目标。乙酰胆碱酯酶（AChE）最广为人知的功能是水解神经递质乙酰胆碱，它通过与淀粉样 β-肽相互作用，在阿尔茨海默病的淀粉样变性中具有一个外周别构亚基。然而，AChE 还具有其他非水解功能。在这里，我们使用计算工具鉴定和表征了 AChE 中的两个新的别构位点，这使我们能够通过基于结构和片段热点策略的虚拟筛选来识别别构抑制剂。鉴定出的化合物还进行了体外抑制 AChE 的筛选，发现有三种具有活性。进一步进行了实验（动力学）和计算（分子动力学）研究来验证别构活性。这些新化合物可能是研究 AChE 非胆碱能功能的有价值的药理学工具。
