囊泡电化学传感器的构建及其在药物-细胞膜亲和力筛选模型的应用研究

Based on the fabrication of vesicle electrochemical sensor, the quantitative relationship between the electrochemical response value and concentration of the sample is established by drawing lessons from the theoretical and quantitative methods of receptor-pharmacodynamics. The parameter of affinity (Kd), which directly reflects the activity of drugs, is obtained by using the related mathematical processing of linear transformation and computer nonlinear fitting. The screening model based on drug-cell membrane affinity is established according to the large amount of experimental data by using the vesicle electrochemical biosensors. At the same time, these experimental and quantitative models are verified by using the drug screening method bsed on the optical biosensor. Then vesicle electrochemical sensor array is investigated in order to realize the rapid drug screening. The core target of the high throughput screening, which performs the international advanced level and independent innovation, will be realized. It will help to offer the powerful technology of domestic drug screening and accelerate the process of researching new drugs with independent property rights in our country.

以构建囊泡电化学传感器为基础，通过借鉴受体-药效学上的理论及其定量方法，建立电化学响应值与样品浓度的量化关系，利用线性变换和计算机非线性拟合进行相关数学处理获得亲和力参数Kd，从而直接反映药物活性。依据大量的实验数据建立基于囊泡电化学传感器研究药物-细胞仿生膜亲和力用于药物活性筛选的实验模型和定量模型，并采用光学传感器药物筛选方法验证其用于药物筛选的可行性，继而研究构建囊泡阵列电化学传感器，实现快速药物筛选，最终实现达到具有国际先进水平的、自主创新的高通量筛选检测模型的核心目标，为国内新药筛选提供有力技术支撑，加快我国具有自主产权新药研发的进程。

以构建囊泡等电化学传感器为基础，通过借鉴受体-药效学上的理论及其定量方法，运用电化学响应值与样品浓度的量化关系，利用线性变换和计算机非线性拟合进行相关数学处理，推导得到了计算药物-细胞膜亲和力测定的定量模型。通过在细胞表面过表达beta2-肾上腺素受体激动剂和CHO-alpha1肾上腺素受体两种靶点，并通过囊泡、纳米金/壳聚糖和聚赖氨酸等材料固定两种靶点，构建了相关电化学传感器，采用电化学阻抗、循环伏安法或示差脉冲伏安法研究了利多卡因、丁卡因、福莫特罗、丙卡特罗和肾上腺素等药物与靶点之间相互作用，通过相关电化学信号的变化可识别药物与靶点之间的作用，并利用推导得到的药物-细胞膜亲和力测定的定量模型计算得到了靶点和药物之间亲和力Kd，初步构建了药物-细胞膜亲和力筛选模型，为药物的快速筛选提供参考。在该项目的资助下，发表相关SCI论文7篇。