Development of methods for in vitro testing of potential drugs against Alzheimer disease

Charles University in Prague Faculty of Pharmacy in Hradec Králové Healthcare bioanalytics - Specialist in Laboratory Methods Department of Biochemical Sciences Candidate: Bc. Martina Hrabinová Supervisor: Doc. PharmDr. Tomáš Šimůnek Ph.D. Supervisor specialist: Pplk. Doc. PharmDr. Daniel Jun Ph.D. Title of diploma thesis: Development of method for in vitro testing of potential drugs Alzheimer disease Alzheimer's disease (AD) is a neurodegenerative disease with increasing incidence. Although many decades have passed since the disease was discovered, there is no causal therapy yet. Currently available therapy consists in treatment with central acetylcholinesterase inhibitors and memantine, improving the patient's quality of life. The aim of this study was to develop a simple colorimetric method for the determination of prolyl oligopeptidase (POP) and beta secretase (BACE) activity, important enzymes associated with AD pathogenesis, and to use this method for screening of potential AD drugs and to determine their antioxidant and antiradical activity. Results showed that tested substances were weak inhibitors of POP compared to standard inhibitor Z- Gly-Pro- prolinal. Standard cholinesterase inhibitors used in AD therapy showed no ability to inhibit POP. The method for determination of BACE activity..