Preparation And Application Of Genechip For Genotyping The Hot Mutations Of Drug Metabolizing Enzyme

Preparation and Application of Genechip for Genotyping the Hot Mutations ofDrug Metabolizing EnzymeCytochrome P450 super family are important enzymes metabolizing a serials of endothelic and xenobiotic chemistry, including many drugs, precacinogen, and pretoxigen, CYP450s involve in the clearation of about 60% clinical drug. The sub-family 1-3 of CYP450 have been studied deeply, more and more concentrate on the relationship between their function and molecular properties. CYP 2C9, CYP 2C19, CYP 2D6 and CYP 3A4 are important to drug metabolism, and CYP 1A1, CYP 1A2, CYP 1B1, CYP 2E1 and CYP 3A4 are necessary to the metabolizing activation of carcinogens. Traditional genotyping technologies hold back the progress of pharmacogenomics. Developing high throughout, fast and accurate genotyping method is the key.Many studies have proven the genetic polymorphisms of CYP450s. Genetic variations contribute to these polymorphisms, influencing the general/adverse drug response and toxicity of individuals. Lots of pathogenic xenobiotics will be activated or inactivated by the polymorphisms; it is depending on the type of catalytic reaction. Sensitivities to special disease maybe decrease or increase when the activity going down. Because of the metabolizing importance and intimate relevance to diseases, so rapid progress were get in studies about CYP450. Base on the enzymatic importance, higher frequencies in Chinese and functional mutation, We chose 8 SNPs from CYP 1A2, CYP 2D6, CYP 2C19, CYP 2C9 and CYP 3A4, investigating the genetic characterizes of Chinese metabolizing enzymes to build independent database of functional metabolism polymorphism ourself, which severing in clinical therapy or drug development.With the microarray producing platform in our center, we designed a set of low density genechip genotyping SNPs of the metabolism enzymes, and accessed tosampling, MultiPCR and products-mixed hybridization have been done, and the resolution was got to genotype metabolism enzymes with oligonucleotide-based genechip. We analysis the genotyping results and pharmacokinetics of CYP 3c9*3 (taking tolbotemide as probe drug) from the same subjects, the CYP 3c9*l/*3 frequency is 6.6%, and individuals with this genotype have a lower metabolizing to tolbotemide. We also sequencing the PCR products from different genotype groups which identified by genechip, the results were confirmed. So we conclude that, after large scale clinical test, it is feasible to develop drug metabolism enzyme genotypingmicroarray with quality control.