Research Of P-Nitro Caffeic Acid Phenethyl Ester In Vitro Metabolism

Caffeic acid phenethyl ester (caffeic acid phenethyl ester, CAPE) as one of the main active ingredient of propolis extract, it has become the hot new drug of research in recent years, it can clear suppression of inflammatory responses, immunomodulatory and anti-tumor, and many other biological activity. Many researchers have carried out studies of the structural modification of the screened compounds have a stronger biological activity than caffeic acid phenethyl ester.Our group innovative synthesis of such derivatives, nitro-caffeic acid phenethyl ester in the2-phenylethyl-3-(3,4-hydroxyphenyl)-2-acrylic phenylethyl bit by adding nitro. The most important feature of this derivative is significantly enhanced compared with caffeic acid phenethyl ester in immune regulation. Not yet in-depth study of caffeic acid phenethyl ester (CAPE) in nitro-regulation of the immune mechanism of action is unclear, so we need to be in-depth exploration, and in vitro metabolism study is a good entry point.The in vitro metabolism of liver in vitro metabolism of the intestinal metabolism of bacteria outside the research, as well as other organs in vitro metabolism studies. This paper referred to in vitro metabolism studies in vitro hepatic drug metabolizing enzyme metabolism of drugs, which is today a very important drug metabolism research tools, but also the metabolism of hot spots. Hepatic drug metabolizing enzyme research methods to animal liver or liver cells to construct the system of in vitro liver metabolism studies, and liver microsomes in vitro incubation method is the most mainstream of the in vitro liver metabolism studies. It has, such as the enzyme preparation technology is simple, fast metabolic processes, results and good reproducibility, experimental study is easy to operate and can also be used for the inhibition of hepatic drug metabolizing enzyme and in vitro metabolic clearance of research advantages. Based on these advantages, the method is widely used in practical work. The in vitro metabolism to rid the body of interference, direct observation of select metabolic enzyme on the substrate to provide a favorable reference data for the subsequent pharmacological experiments.In vitro metabolism studies can not be separated from the enzymatic reaction kinetics (kinetics theory of by enzyme-catalyzed reactions), it is to support the cornerstone of in vitro metabolism study. Simply put, the enzymatic reaction kinetics of enzymatic reactions in the enzymatic reaction rate and its influencing factors, such as enzyme concentration, substrate concentration, pH of the reaction system, the temperature of the reaction system, inhibitors and activation the type and concentration of the agent. Study the end of the concentration on the reaction rate, Michaelis-Menten equation is essential for the theoretical basis. The so-called Michaelis-Menten equation (the Michaelis-Menten equation) is deduced in1913by German chemist Michaelis and Menten mathematical expressions that represent the basic principles of enzymatic kinetics (Michaelis-Menten equation:v=Vmax of [S]/(Km of+[S])) This equation indicates that the quantitative relationship between the substrate concentration [S] and the enzyme reaction rate [V], to better reveal the relationship between drugs and metabolism.In this paper, the following main points:1. Liver drug metabolizing enzymes involved in the metabolism of nitro caffeic acid phenethyl ester in rat liver microsomes in vitro enzyme kinetics of the metabolism process;2.inhibition types of metabolic inhibitors;3. metabolites;4. provide the basis for rational drug use in combination drug interactions and clinical safety.Use of:first, the preparation of liver microsomes; then incubated metabolism, the use of rat liver microsomal suspension concentration is diluted to0.08g1-1suspension, while the metabolism of the original drug concentration in vitro metabolic experiments vary; Finally, sample handling, after the end of the reaction by adding three times the amount of cold acetonitrile and vortex mixed to terminate the reaction at4℃,10000g centrifugation for10min, the supernatant, NaCl60-70mg of Vortex for2min,10,000g of centrifugation for5min, pipet the upper liquid in the centrifuge tube, and then at50℃water bath evaporated to dryness the residue obtained is dissolved in acetonitrile,0.22μm microporous membrane, taking into20μL samples. Will nitro caffeic acid phenethyl ester to join the rat liver microsomal incubation, and then detect the remaining amount and use of the theory of enzyme kinetics data analysis.The experimental results show that the nitro caffeic acid phenethyl in rat liver microsomes body mainly by the CYP450enzyme system (cytochrome P450) in the CYP4503A4and the CYP4502C19these two enzymes metabolized, while ketoconazole and Lansuo of omeprazole can be a significant anti-competitive inhibition of metabolism in rat liver microsomes, this feature can provide the basis for rational drug use in combination drug interactions and clinical safety. The major metabolites of the nitro-caffeic acid phenethyl ester, nitro-beta-phenylethyl alcohol, caffeic acid, ferulic acid and nitric Jia Wei acid phenethyl ester, in which the metabolic formation of caffeic acid, ferulic acid is highly group of biologically active molecules, so you can commence from here on the nitro caffeic acid phenethyl immunomodulatory mechanism.