Association Between The CYP2C9 Polymorphism And Metabolism Of Glibenclamide

【Objective】Study of the CYP2C9 gene polymorphism in healthy controls and diabetics; Analysis the effect of the human body CYP2C9 enzyme mutant allele on glibenclamide drug metabolism. Elucidate the clinical treatment of drug reaction individual differences from the perspective of individual genetics, to provide a evidence for guide glibenclamide and other diabetes drugs clinical rational application , to achieve diabetes individuation medical ,and to lay the foundation for the Pharmacogenetics study.【Methods】Polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) and sequencing methods were used to study the CYP2C9 gene polymorphism on promoter region,exon 2,exon 3,exon 4,exon 6 and exon 7 mutational site and frequency analysis in 150 healthy controls,100 typeⅡdiabetics and 7 cases of long-term use of glibenclamide treatment of typeⅡdiabetics;HPLC techniques were used to study the drug metabolism for 7 cases of long-term use of glibenclamide treatment of typeⅡdiabetics after 1,2.5,4,8h at different time points. 3P97 pharmacokinetic software was used to calculate pharmacokinetic parameters and paired t-test to comparison.【Results】1. In the detection of 150 healthy controls, CYP2C9 gene promoter region -1188C/T sites,exon 2 269T>C sites,exon 3 430C>T sites,exon 6 895A>G sites and exon 7 1075A>C sites were 53.9% 1.3%,0%,2%,7% heterozygous carriers; 100 diabetics in these sites were 60.4%,0%,1%,1%,7% heterozygous carriers . Exon 4 found no polymorphism loci.2. Statistical analysis showed that CYP2C9 gene promoter region -1188C/T,exon 2 269T>C and exon 3 430C>T mutational site exist quite different, but no statistical significance . The present study results with the other parts of China in total population, the Asian population( Japanese and Korean), Europe and the United States population CYP2C9 allele frequency comparisons found : exon 3 430C>T sites,with the population Europe and the United States, there was significant difference (Χ2p =65.3154, P<0.05); exon 7 1075A>C sites with the Asian population ( Japanese and Korean) (Χ2p =5.6488. P <0.05) and Europe and the United States population (Χ2p =11.6795. P <0.05) were significantly different, Other results of the comparison sites no significant difference (P>0.05).3. In glibenclamide drug metabolism experiment found one poor metabolizer(PM). CYP2C9 gene sequencing showed that the subject of PM is a exon 6 895A>G site CYP2C9*1/*16 heterozygous genotype. The subject of PM glibenclamide plasma half-life of 51.87h. and other diabetics subjects (CYP2C9*1/*1) on average half-life of the individual 5.775h. It is about 9 times than CYP2C9*1/*1 genotype.【Conclusion】1. Polymorphism study showed, CYP2C9 gene promoter -1188C/T sites. Exon 2 269T>C sites, exon 3 430C>T sites, Exon 6 895A>G sites and exon 7 1075A>C sites there are polymorphic loci. Including 150 healthy controls and 100 typeⅡdiabetics these sites were 56.5%,0.8%,0.4%,1.6%,7.0% heterozygous carriers,and in exon 4, we found no polymorphism loci.2. The present study results with the other parts of China population, the Asian population( Japanese and Korean), Europe and the United States population CYP2C9 allele frequency comparisons found: exon 3 430C>T sites, there was significant difference with the population Europe and the United States;exon 7 1075A>C sites,there was significant difference with the Asian population ( Japanese and Korean), Europe and the United States population(P<0.05).3. Diabetics with glibenclamide metabolism study shows that CYP2C9 enzyme mutant allele CYP2C9*16(Exon 6 895A>G sites)to glibenclamide Pharmacokinetics significant affected, The research show that according to CYP2C9 gene typing of clinical specimens and Individual medical is of great guiding significance.