Construction And Application Of Alpha 1A-adrenocepter Biochromatographic Model

Active components screening in complex system is a hot field of pharmacology, and is also an important way to find original new drugs. The active component screening methods include:animal model selection, solution-cells selection, enzymes and receptor model selection and classic chromatography model selection. The establishment and application of these models play a positive role for drug screening of active components, but these methods need to be further improved in the specific areas. This paper is mainly focus on the research of construction and application of alpha 1A-adrenocepter (α1A-AR) biochromatographic model, and aim to establish a new screening method for the active component. The paper has three chapters. The author's main contributions are as follows:1. Based on existent methods of screening active component, the author make use of the high separate capacity of chromatography in complex system and the high specificity of adrenocptors to propose a new model of biochromatographic screening of drug active ingredients.2. The author used the method of cell culturing and affinity chromatography to obtain alpha 1A-adrenocepters. Then, receptors were immobilized on the surface of macro porous silica gel by chemical coupling method, and the bonding sites and the bioactivity of the receptor on the surface of the stationary phase were characterized. Based on these, the biocharomatographic model was implicated to screening active components of Safflower. The result show that, capacity factors of terazosin, noradrenaline, tamsulosin, aramine and urapidil respectively are 3.785,5.260,6.065,4.512 and 3.275. Thus proves that the alpha 1A-adrenocepters biocharomatographic model can be used to screening active components of complex system.3. The number of binding sites, association constant and influencing of immobilization alpha 1A-adrenocepters with prazosin and terazosin were determinated by the methods of frontal analysis and zonal elution. The result show that, the association constant of prazosin with alpha 1A-adrenocepters on the biochromatographic column was 1.6×105M-1, the binding sites was 5.8×10-6M. The main force on the binding process between prazosin and alpha 1A-adrenocepters was the electrostatic force. Terazosin and alpha 1 A-adrenocepters has two types of binding sites, the total binding sites was 6.1 x 10-6M, association constants were 1.1×105M-1,0.1 x 104M-1, The main force on binding process between prazosin and alpha 1A-adrenocepters was hydrophobic interaction. All the above results demonstrated that the receptor biocharomatographic model was a new technology for online investigating interaction between receptor and drug.