Investigation On The Interaction Between Five Active Components Of Chinese Traditional Herb And Human Serum Album By High-performance Affinity Chromatography

The study on the interaction between small molecule, especially multi-component molecules of Chinese traditional herb and proteins is very important. It is expected to provide a theoretical basis and guidance for the modernization of traditional Chinese medicine, such as the molecular pharmacology of Chinese medicine, Chinese medicine active ingredients screening, transforming the molecular structure of drugs, the scientific understanding of traditional Chinese medicine compound, as well as drug compatibility. In this thesis, the interaction between five active components of Chinese traditional herb and human serum album (HSA) immobilized onto chromatographic supports had been investigated by high-performance affinity chromatography with frontal analysis and competition and displacement studies. The obtained results are as follows:1. The adsorptions of these five active components of Chinese traditional herb on the immobilized HSA column were determined by frontal analysis. Two kinds of binding isotherm models were used to judge the types of binding site of these drugs on HSA. The results showed that there was only a single type of binding site on HSA for ferulic acid, paeonol, chlorogenic acid and vanillic acid, and two types of binding site for puerarin.2. The self-competition approach of competition and displacement studies was also used to determin the binding constants of five active components of Chinese traditional herb. It was found that the interaction between HSA and the five drugs was of weak interaction.3. The direct competitive approach of competition and displacement studies was used to study the direct interaction between three drugs, ferulic acid, paeonoT and puerarin. The conclusion that ferulic acid, paeonol and puerarin competed with each other at the same type of binding site of HSA molecular can be used to explain the Chinese medicine theory, in which each drug's clinical efficacy can be improved by utilizing the compatibility of Chinese traditional medicine. Moreover, ferulic acid and paeonol had a direct competition at the same type of binding site of HSA molecular which was indole site (site II) by using probe reagents.4. The effects of temperature on the interaction between four active components of Chinese traditional herb and immobilized HSA were investigated. According to the thermodynamic parameters, the main type of binding force between these drugs and HSA was suggested. The results showed that the main sort of binding force was hydrogen bonding and van der waals force for ferulic acid and paeonol binding with HSA, and electrostatic force for chlorogenic acid and vanillic acid.