Analitičke metode za praćenje metaboličke aktivnosti humanih citokrom P450 enzima

master's thesis

• Title (english): Analytical methods for assessing the metabolic activity of human cytochrome P450 enzymes
• Author: Dominik Dautović
• Mentor: Mirza Bojić (mentor)
• Committee member: Mirza Bojić (predsjednik povjerenstva)
• Committee member: Hrvoje Rimac (član povjerenstva)
• Granter: University of Zagreb; Faculty of Pharmacy and Biochemistry; (Department of medicinal chemistry); Zagreb
• Defense date and country: 2020-10-28, Croatia
• Scientific / art field, discipline and subdiscipline: BIOMEDICINE AND HEALTHCARE; Pharmacy; Pharmacy

Abstract

Citokrom P450 (CYP) je superporodica enzima koja ima ključnu ulogu u metabolizmu lijekova i brojnih ksenobiotika uvođenjem hidrofilnih skupina u njihove molekulske strukture. Glavni CYP enzimi uključeni u metabolizam lijekova su CYP3A4, CYP2D6, CYP2C9, CYP1A2 i CYP2C19. Genetski predodređene razlike ili utjecaj vanjskih čimbenika mogu uzrokovati promjenu enzimske aktivnosti citokroma P450, što može dovesti do različitih učinaka, uključujući neželjene i štetne reakcije lijekova ili gubitak terapijskog djelovanja. Ovaj rad daje pregled i usporedbu najčešće korištenih metoda za praćenje metaboličke aktivnosti humanih citokrom P450 enzima. Opisan je opći tijek postupaka analize, te prednosti i ograničenja metoda koje se temelje na tekućinskoj kromatografiji visoke djelotvornosti (HPLC), koja je najčešće spregnuta sa spektrometrijom masa (MS), a zatim i metoda koje se temelje na fluorescenciji, luminescenciji ili radioaktivnosti. Prikazana je njihova primjena u in vitro i u in vivo praćenju metaboličke aktivnosti humanih citokrom P450 enzima uz specifične marker supstrate ili inhibitore, koja su provedena u svrhu fenotipizacije radi optimizacije terapijskih ishoda, utvrđivanja odnosa fenotipa i genotipa, probira modulacijskih učinaka kandidata za nove lijekove, proučavanja inhibicijskog ili induktivnog djelovanja lijekova, ljekovitog bilja, sastojaka hrane i prehrambenih aditiva, te usporedbe postojećih i validacije novih pristupa.

Abstract (english)

Cytochrome P450 (CYP) is a superfamily of enzymes that plays a key role in the metabolism of drugs and numerous xenobiotics by introducing hydrophilic groups into their molecular structures. The major CYP enzymes involved in drug metabolism are CYP3A4, CYP2D6, CYP2C9, CYP1A2, and CYP2C19. Genetically determined differences or modulation of cytochrome P450 enzyme activity can lead to a variety of effects, including adverse drug reactions or loss of therapeutic effects. This paper provides an overview and comparison of the most commonly used methods for assessing the metabolic activity of cytochrome P450 enzymes. The general work flow of these methods is described, as well as the advantages and limitations of each method. These methods include high performance liquid chromatography (HPLC)-based methods, which are most often coupled with mass spectrometry (MS); fluorescence-based; luminescence-based; and radioactivity-based methods. Various studies where these methods have been applied for in vitro and in vivo evaluation of cytochrome P450 metabolic activity using specific markers are presented in this paper. Numerous activity measurements have been performed: for the purpose of phenotyping cytochrome P450 enzymes to optimize therapeutic outcomes; to determine the relationship between phenotype and genotype; for screening of modulatory effects of drug candidates and studying the inhibitory or inductive effects of drugs, herbs, food ingredients and additives; and also for comparisons of existing approaches and validation of new approaches for cytochrome P450 activity assessment.

• Keywords: citokrom P450; metabolizam lijekova; metabolička aktivnost
• Keywords (english): cytochrome P450; drug metabolism; metabolic activity
• Language: croatian
• URN:NBN: urn:nbn:hr:163:008276
• Study programme: Title: Pharmacy; Study programme type: university; Study level: integrated undergraduate and graduate; Academic / professional title: magistar/magistra farmacije (magistar/magistra farmacije)
• Type of resource: Text
• File origin: Born digital
• Access conditions: Open access
• Created on: 2021-04-23 13:40:55