Active ingredients of beta _2- adrenergic receptor chromatographic screening in Maxingshigan Decoction
|Keywords||β2-adrenoceptor Biochromatography Screening Maxing Shigan Decoction Bioactive compounds|
Major diseases which are prone to happen is a main factor which is harmful to human’s health and the quality of life. As an effective means to prevent and treat diseases, to research efficient and innovative drugs is an eternal hot issue of chemistry, biology and pharmaceutical field. Although computer-aided drug design, drug molecules’discovery based on molecular fragments, high-throughput drug screening methods have been successfully used in drug discovery and development process, but the number of new drugs in international markets have shown a downward trend in recent years. The essential reason is that the above methods are not fundamental solutions to screening the active pharmaceutical ingredients for low success rate and long life cycle. Thus construction of efficient screening methods for active pharmaceutical ingredients is still one of the key scientific problems to be solved in medicine field. In this thesis, we choose one of the traditional Chinese medicine Maxing Shigan decoction, to conduct a systematic study of high-throughput screening its active ingredients. Designing to provide ideas for the receptor chromatography to screening of the active ingredients in traditional Chinese medicine. This paper is divided into four chapters, the author’s main contributions are as follows:1. Established high performance liquid chromatography-electrospray ionization-time of flight tandem four-pole detection method to detect ephedrine of Maxing Shigan decortion; Orthogonal design was employed to optimize the engineering for the extracting procedure of ephedrine. The results showed that the peak area and the concentration of ephedrine gave a linear relationship during the range of0.005-1.0mg/mL, precision determination of the relative standard deviation is0.28%, reproducible determination of the relative standard deviation is0.51%, the average recovery is95.4%. It demonstrated that this method can be used to determining ephedrine in ephedra and its compound preparation; the optimized engineering of ephedrine was believed to be the process of30fold water, a value of 80℃for heating temperature, a repeatability of4times and40min for each time. It can be applied to use β2-adrenergic receptor chromatography to screening the active ingredients of Maxing Shigan decortion and provide stable and reliable sample preparation and quality control methods.2. The bioactive compounds of Maxing Shigan decoction were screened by β2-adrenoceptor biochromatography, use the competitive displacement analysis method to study the interaction between receptor and the active ingredients. The result showed that ephedrine is the active ingredient in Maxing Shigan decortion which had the specific interaction with β2-AR, The association constant of ephedrine with β2-AR in the biochromatographic column was5.85×103/M, and the binding sites were1.72×10-7M. It proved that the receptor chromatography can be used for rapid screening of specific active ingredients in traditional Chinese medicine and its receptor interaction mechanism.