Selection Of Dihydrofolate Reductase Inhibitors And Enzyme’s Immobilized

Dihydrofolate reductase (dihydrofolate reductase, EC1.5.1.3) is a key enzyme inthe metabolism of nucleic acid, it catalyze transformation of dihydrofolate intotetrahydrofolate, while folate participate in the synthesis of DNA and RNA in thepurine nucleotides metabolism, it can promote the proliferation of cells lysed. Tocontrol the generation of tetrahydrofolate become the main research for anti-cancerdrugs. The dihydrofolate reductase become an important target enzyme for anticancerdrugs. Dihydrofolate reductase inhibitors screened by UV spectrophotometry, and theimmobilized anti-cancer drug screening model was established.Twelve artificial inhibitors were chosen. The dihydrofolate reductase determinedby UV spectrophotometry for inhibitors, through data comparison of CM2, BP1, BP7,CA9, BPF1, BPF2, HS11etc. inhibitors, the corresponding decision coefficients were:0.8507,0.8109,0.9931,0.9671,0.9779,0.9651,0.9434. It will affect as inhibitortogether with other CI5、CI8、CI9、BPF3、TV95enzymes.Dihydrofolate reductase were extracted from human liver, the inhibitor screeningfrom experiments were tested, the results indicated that BPF1, CA9either in animalexperiment or in this experiment the correlation coefficient is maintained above0.9respectively:0.9873,0.9640. While the activity of CA9is weak, so determine theBPF1as inhibitor to human dihydrofolate reductase.Immobilization condition of dihydrofolate reductase and stability were explored,analysis cross-linking embedding and embedding cross-linking were analysed, theresults showed the that the latter activity was higher than the control and reached apeak when the concentration is4%glutaraldehyde, so choose the embeddingcross-linking immobilized dihydrofolate reductase. Build BPF1-immobilizedenzyme model, measured obtaining the coefficient of determination was0.9523,prove inhibitor effect is still good. This was provided make dihydrofolate reductase asnew anticancer drugs in vitro assay may be possible.