Ispitivanje utjecaja derivata itakonske kiseline na koncentraciju tiolnih skupina ljudskoga seruma

master's thesis

• Title (english): Examining the effect of itaconic acid derivates on thiol group concentration in human serum
• Author: Luka Mavrić
• Mentor: Karmela Barišić (mentor)
• Committee member: Karmela Barišić (predsjednik povjerenstva)
• Committee member: Roberta Petlevski (član povjerenstva)
• Committee member: Lovorka Vujić (član povjerenstva)
• Granter: University of Zagreb; Faculty of Pharmacy and Biochemistry; (Department of medical biochemistry and haematology); Zagreb
• Defense date and country: 2021-03-26, Croatia
• Scientific / art field, discipline and subdiscipline: BIOMEDICINE AND HEALTHCARE; Pharmacy; Medical Biochemistry

Abstract

Unatoč prednostima lijekova koji učinak postižu kovalentnim vezanjem za ciljne molekule, rizik od toksičnosti uzrokovane nespecifičnim vezanjem predstavlja komplikacije u njihovom razvoju. Neki od tih lijekova su Michaelovi akceptori. Radi se o elektrofilima koji se kovalentno vežu za cisteinske ostatke ciljnih proteina. Osim toksičnosti zbog promjene svojstava vezanih proteina, nespecifično kovalentno vezanje Michaelovih akceptora za tiole proteina i peptida dovodi i do deplecije glutationa. Deplecijom glutationa narušava se funkcija mehanizama antioksidacijske obrane i dolazi do stanja oksidacijskog stresa. U ovom radu ispitan je utjecaj jedanaest potencijalnih citostatika na koncentraciju tiolnih skupina ljudskog seruma. Ispitani spojevi derivati su itakonske kiseline i Michaelovi akceptori. Ispitan je i utjecaj antimalarika koji su strukturne komponente nekih derivata. Ispitani spojevi dodani su u uzorke ljudskog seruma te je nakon inkubacije od sat vremena određena koncentracija tiolnih skupina. Svaki spoj ispitan je uz inkubaciju pri temperaturi od 37 °C i uz inkubaciju pri sobnoj temperaturi. Za određivanje koncentracije tiolnih skupina korištena je metoda s Ellmanovim reagensom. Izraženo smanjenje koncentracije tiolnih skupina ukazuje na nespecifično vezanje ispitanog spoja. Od jedanaest ispitanih derivata itakonske kiseline, samo dva derivata, spojevi V9 i MI9, nisu uzrokovali pad koncentracije tiolnih skupina veći od 30% ni pri jednoj temperaturi. Nijedan od ispitanih antimalarika nije uzrokovao pad koncentracije tiolnih skupina veći od 10% ni pri jednoj temperaturi. Za detaljniju karakterizaciju in vivo opterećenja nespecifičnim kovalentnim vezanjem ispitanih derivata itakonske kiseline potrebno je provesti dodatna ispitivanja.

Abstract (english)

Although the drugs that achieve their effect by covalently binding the target molecules have their advantages, the risk of toxicity caused by nonspecific binding poses complications in their development. Some of those drugs are Michael acceptors. They are electrophiles that covalently bind cysteine residues of target proteins. In addition to toxicity caused by the modification of bound protein's properties, nonspecific covalent binding of Michael acceptors to protein and peptide thiol groups leads to glutathione depletion. Glutathione depletion damages the function of antioxidant defense mechanisms and causes the state of oxidative stress. In this paper, the effect of eleven potential cytostatics on the concentration of thiol groups in human serum was tested. The tested compounds are itaconic acid derivatives and Michael acceptors. Antimalarics that are structural components of some derivatives were also tested. The tested compounds were added to the samples of human serum. Thiol group concentration was determined after a one-hour incubation. Each compound was tested with an incubation at 37 °C and with an incubation at room temperature. Thiol group concentration was determined using the Ellman reagent method. Substantial decrease in thiol group concentration indicates nonspecific binding of the tested compound. Out of eleven tested itaconic acid derivatives, only two compounds, V9 and MI9, didn’t cause a decrease in thiol group concentration greater than 30% at either temperature. None of the tested antimalarics caused a decrease in thiol group concentration greater than 10% at either temperature. Additional testing is required for further characterization of in vivo nonspecific covalent binding burden of the tested itaconic acid derivatives.

• Keywords: itakonska kiselina; derivati itakonske kiseline; Michaelovi akceptori; nespecifično kovalentno vezanje; Ellmanov reagens
• Keywords (english): itaconic acid; itaconic acid derivatives; Michael acceptors; nonspecific covalent binding; Ellman reagent
• Language: croatian
• URN:NBN: urn:nbn:hr:163:930090
• Study programme: Title: Medical biochemistry; Study programme type: university; Study level: integrated undergraduate and graduate; Academic / professional title: magistar/magistra medicinske biokemije i laboratorijske medicine (magistar/magistra medicinske biokemije i laboratorijske medicine)
• Type of resource: Text
• File origin: Born digital
• Access conditions: Open access
• Created on: 2021-11-16 16:16:27