|
|
Development of methods for in vitro testing of potential drugs against Alzheimer disease
Hrabinová, Martina ; Šimůnek, Tomáš (advisor) ; Soukup, Ondřej (referee)
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...
|
|
| |
|
|
Development of methods for testing drugs affecting the CNS
Hrabinová, Martina ; Jun, Daniel (advisor) ; Patočka, Jiří (referee)
1 Abstract and keywords The current thesis aims at the production of three enzymes, including beta-secretase 1 (BACE1; beta-site amyloid precursor protein cleaving enzyme 1), acetylcholinesterase (AChE), and butyrylcholinesterase (BChE). BACE1 is an integral membrane protein that plays a crucial role in the amyloid precursor protein's cleavage. The product is subsequently processed by y-secretase, producing amyloid-beta peptides and insoluble amyloid plaques in Alzheimer's patients (AD). According to the Annual Report of the Czech Alzheimer's Society, 158,000 patients were diagnosed in 2019. By 2050, this number is supposed to reach 300,000 patients. AChE inhibitors and N-methyl-D-aspartate receptor antagonists are currently the only alternatives for AD therapy. AChE is also a target enzyme in nerve agent (NA) poisoning (together with BChE). The Department of Toxicology and Military Pharmacy focuses on studying NA effects and medical protection against them. From this perspective, the production of AChE and BChE is essential for developing and evaluating newly synthesized cholinesterase (ChE) reactivators and inhibitors. The thesis focuses on introducing the expression system Expi293 to produce human recombinant enzymes BACE1, AChE, and BChE. For each enzyme, the individual steps required to obtain a...
|
|
|
Development of methods for in vitro testing of potential drugs against Alzheimer disease
Hrabinová, Martina ; Šimůnek, Tomáš (advisor) ; Soukup, Ondřej (referee)
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...
|
|
| |
guest ::
RSS feed.