Dynamin Inhibitors as Tools for Dissecting the Endocytic Pathway in Neurons

神经细胞通过释放神经递质进行交流，神经递质被包裹在神经末梢内的突触小泡中。囊泡的数量是有限的，因此它们被回收(内吞作用)以供重复使用。一些人类神经疾病劫持了病理性多肽、蛋白质或病毒进入细胞内的途径，从而瘫痪、杀死或感染神经元。我们的总体目标是控制神经沟通，最终使我们能够治疗癫痫等神经沟通障碍。在最极端的情况下，完全阻断内吞作用会迅速导致神经通讯完全阻断。因此，减缓(而不是阻断)可能是控制某些神经疾病的一种手段。例如，癫痫发作是神经元不受控制的放电。控制内吞作用的主要机制集中在蛋白质动力素上。动力素可以组装成一个微小的、紧紧缠绕的螺旋或弹簧。当能量(GTP水解)被施加到纳米弹簧上时，它会迅速释放，从细胞壁分离出空的再循环突触小泡，回到神经元中。我们的前提是，阻止纳米弹簧可能会导致新一代抗癫痫药物的出现。为了实现这一点，我们已经发现了第一种动力蛋白的化学抑制剂。在这个项目中，我们将通过展示它们针对Dynamin中不同的位置来确定它们是如何工作的。我们已经开始了一项雄心勃勃的化学合成计划，以极大地提高抑制剂的效力和特异性。我们将使用组合化学的迭代方法来扩展这一点。当应用于神经元时，这些药物似乎是第一批内吞抑制药。将测试我们的提议，即他们将揭示动力蛋白在内吞作用的不同阶段的多个作用点。该项目将证明抗动力素药物的开发可能导致第一个抗内吞药物的原理。这有可能导致未来靶向抗癫痫和抗癌药物的开发。

Nerve cells communicate by the release of neurotransmitters which are packaged in synaptic vesicles inside nerve endings. There is a finite number of vesicles, so they are recycled (endocytosis) for reuse. Some human neural diseases hijack the endocytic pathway for entry of pathological peptides, proteins or viruses to paralyse, kill or infect neurons. Our overall aim is to control nerve communication to ultimately allow us to treat disorders of nerve communication like epilepsy. At its most extreme, completely blocking endocytosis quickly results in a complete block in nerve communication. Therefore slowing it down (rather than blocking) might be a means to control some neural diseases. For example, a seizure is the uncontrolled firing of neurons. The main mechanisms controlling endocytosis converge on the protein dynamin. Dynamin can assemble into a tiny, tightly wound helix or spring. When energy (GTP hydrolysis) is applied to the nanospring it rapidly releases to cleave off empty recycling synaptic vesicles from the cell wall back into the neuron. Our premise is that blocking the nanospring may lead to a new generation of antiepileptic drugs. To achieve this we have already discovered the first chemical inhibitors of dynamin. In this project we will determine how they work, by showing that they target distinct sites in dynamin. We have embarked on an ambitious chemical synthesis program to greatly improve the potency and specificity of the inhibitors. We will expand this with an iterative approach using combinatorial chemistry. When applied to neurons, the drugs appear to be the first endocytosis inhibitors. Will test our proposal that they will reveal multiple points of action of dynamin in various stages of endocytosis. This project will prove the principle that the development of anti-dynamin drugs could lead to the first anti-endocytic drugs. This has the potential to lead to future development of targeted antiepileptic and anticancer drugs.