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The Application and Study of the Interaction Mechanism between Biomacromolecule and Pharmaceutical Molecules by Fluorescent Probe

Author: ZhaoHu
Tutor: WangHuaiYou
School: Shandong Normal University
Course: Analytical Chemistry
Keywords: Evans Blue Bovine serum albumin Gentamicin Vancomycin Ampicillin Erythromycin Fluorescence Quenching Non-radiative energy transfer Competitive responses
CLC: R96
Type: Master's thesis
Year: 2008
Downloads: 355
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Abstract

The mechanism and the mechanism of the binding between the drug and the biological macromolecule is an important component in the field of pharmaceutical science. Binding site and the mechanism of action of the drug in the human body has a direct impact on the therapeutic effect of the drugs on the disease. Therefore, the study of drugs and biological macromolecules in the the simulated human environment under the conditions of the binding site and mechanism of action is very necessary, can also ensure the safety of people's medication, reasonable and effective. Spectrophotometry in organic analysis and life science research in recent years has shown a rising trend. From the published literature, the application of organic chromogenic reagent from the field of inorganic analysis extended to the the life active substances (antibodies, nucleic acids, proteins, amino acids, etc.) and morphological analysis of environmental pollutants. However, some drugs and biological macromolecules experimental phenomenon or phenomena is not obvious, can not be directly measured or derivation of the experimental data. Fluorescent probe method can better solve this problem, the fluorescent probe method is added drug probe molecular role biological macromolecules or fluorescence of the probe molecule to study the interaction between the drug molecules and biological macromolecules change mechanism, to further understanding of the mechanism of action of the drug molecules in the body. Efficacy of drugs and their adverse two contradictory aspects, how to improve the therapeutic index of the drug, is a closely watched issue. Targeted drug research carried out in recent years, will have a certain effect of the drug with a suitable carrier, by chemical methods combine targeted drug, according to the characteristics of some of the physiological functions of the body, a selective drug delivery to the treatment site. Thereby increasing the therapeutic effect of the drug, reducing the adverse reaction due to systemic distribution of drugs. Most of the currently used by the carrier of endogenous or synthetic macromolecules, such as albumin, antibodies, glycoproteins, dextran, or poly amino acids, etc.. Have special requirements in the application, such as solubility, stability, purity, inherent toxicity, biodegradability, antigenicity, and the degree of difficulty in combination with small molecule drugs. In recent years, molecular fluorescent probes have been widely used in the surface-induced conformational change occurs at the contact surface of biological macromolecules such as studies of protein. Look from a biological pharmaceutically perspective, the serum albumin is the most abundant protein in the plasma, which can in conjunction with many endogenous and exogenous compounds, play a role in the storage and transhipment, is one of the most important biological functions of albumin its ability to transport the drug; characterization albumin binding capacity and the binding point experiments. The characterization process is indirect analysis method based on direct observation of albumin and other substances combined physical and chemical properties change occurs, the direct analysis method; separation of albumin and other substances or conjugates observed changes in physical and chemical properties of albumin occurred . Direct analysis, fluorescence analysis method is widely used and is considered to be superior to indirect analysis method (such as the balanced dynamic permeation method, ultrafiltration method, gel filtration method), this is because it does not destroy binding equilibrium, the closest real reaction. In this thesis, the use of a fluorescent probe method to study the binding site and mechanism of action of the drug molecules and biological macromolecules, so as to further the understanding and the mechanism of action of the study drug in the body and the way to play a pharmacodynamics. Mainly using the fluorescence change caused by the binding between the biological macromolecules, fluorescent probe molecules and drug molecules to study the binding constants of the reaction, binding site, the binding points and the energy transfer, and other parameters, and thus the mechanism of action between them. The first chapter of Application and Prospect of the role of research and application, and the fluorescent probe method in the field of drug molecules and biological macromolecules. The second chapter Evans Blue (Evans blue, EB) fluorescence quenching effect of bovine serum albumin (Bovine serum albumin, BSA) using fluorescence spectroscopy and UV - visible absorption spectroscopy study of Evans blue the quenching bovine serum Albumin reaction. When using 264nm excitation, bovine serum albumin, the maximum emission wavelength at 340nm. Evans Blue and bovine serum albumin interaction, bovine serum albumin fluorescence quenching study, according to the Stern-Volmer equation and the fluorescence lifetime of the types and mechanisms of fluorescence quenching, the quenching mechanism is static quenching, namely Iraq Evans blue and bovine serum albumin to form a stable complex. Experiments seeking to forget the Evans blue and bovine serum albumin binding constants KBSA-EB = 1.122 × 106Lmol-1, and the combination of points n = 0.994. According to the F? Rster non-radiative energy transfer theory, respectively Evans Blue bovine serum albumin efficiency of energy transfer between 0.276 and distance of 3.14nm. Meanwhile, according to the synchronous fluorescence spectra fluorescence derived bovine serum albumin is mainly caused by a tryptophan residue. Finally, the experiments prove that the quenching reaction between the Evans blue and bovine serum albumin and quenching mechanism is energy transfer. The thesis Chapter VI Chapter Evans Blue as a fluorescent probe to study gentamicin (. Gentamicin) bovine serum vancomycin (vancomycin), ampicillin (ampicillin), erythromycin (with erythromycin) white protein of competing reactions. Evans Blue and Bovine Serum Albumin, bovine serum albumin fluorescence quenching according to the Stern-Volmer equation and the fluorescence lifetime of study types and mechanisms of fluorescence quenching. The results show that the quenching type is static quenching, Evans Blue and bovine serum albumin to form a stable complex. Evans blue with bovine serum albumin binding constants, combined with a number of points, and to determine the thermodynamic constants and types of interaction force between EB and BSA. When gentamicin, vancomycin, ampicillin, erythromycin were added, the relative fluorescence intensity of bovine serum albumin recovery. This suggests that these drugs with Evans blue bovine serum albumin occurred competitive reaction. The competitive reaction mechanism of Evans blue and gentamicin drug binding constants were calculated.

The Application and Study of the Interaction Mechanism between Biomacromolecule and Pharmaceutical Molecules by Fluorescent Probe

Abstract

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