Sinteza različnih fluoroforov z azidno ali alkinsko skupino in preučevanje vpliva temperature na intenziteto njihove emisije

Močnik, Anže (avtor), Pajk, Stane (mentor)

Temperatura vpliva na praktično vse biokemične procese. Prilagoditev na temperaturo okolja je ključna lastnost, ki je omogočila preživetje in razvoj živih bitij. Temperatura v celici in njeni okolici je temeljni in izredno pomemben fizikalni parameter za celične sisteme in njihove naloge, kot so delitev celic, difuzija, metabolizem, diferenciacija, izražanje genov in aktivnost encimov. Vpliv zdravil, bolezni, staranja in drugih fizioloških sprememb v celici prek spremembe temperature bi lahko v realnem času spremljali s pomočjo fluorescence. Pojav fluorescence je oddajanje svetlobe iz snovi, do katerega pride, ko snov obsevamo s svetlobo krajše valovne dolžine; snov absorbira svetlobo, ki povzroči prehod elektrona v vzbujeno stanje. Ob vračanju v osnovno stanje spojina nato odda foton višje valovne dolžine od svetlobe, ki jo uporabimo za obsevanje. Spojine, ki so sposobne fluorescence, imenujemo fluorofori. Intenziteta fluorescence nekaterih fluoroforov je odvisna od temperature in različne intenzitete predstavljajo dobro izhodišče za razvoj sond za merjenje temperature na mikroskopskem nivoju (npr. v organelih celic). V tej magistrski nalogi smo sintetizirali različne fluorofore, katerih fluorescenca je odvisna od temperature okolja. Načrtovali smo fluorofore iz družine antracenov, ksantenov in kumarinov, ki smo jih tudi uspešno sintetizirali. Cilj je bil pripraviti vodotopne sonde na osnovi različnih osnovnih skeletov, da bi dobili sonde z različno odzivnostjo fluorescence na temperaturo. Na omenjene fluorofore smo uvedli alkinsko ali azidno skupino, ki omogočata enostavno »klik« kemijo. Po uspešnem sinteznem delu smo izbranim sondam pomerili ekscitacijski in emisijski spekter, na podlagi katerega smo se lahko odločili za ustrezne spojine za nadaljnje preiskave. Izbranim spojinam smo nato pomerili še emisijski spekter v odvisnosti od temperature in ustrezne pare združili s klik reakcijo. Za nadaljnje delo predlagamo še sintezo enakih osnovnih skeletov z drugimi substituenti, za zmanjšanje lipofilnosti, izboljšanje stabilnosti in vodotopnosti.

• Jezik: Slovenski jezik
• Ključne besede: temperatura celice, fluorescenca, fluorofor, temperaturna odvisnost fluorescence, klik reakcija
• Vrsta gradiva: Magistrsko delo/naloga
• Organizacija: FFA - Fakulteta za farmacijo
• Leto izida: 2023
• PID: 20.500.12556/RUL-153166
• Datum objave v RUL: 20.12.2023

MOČNIK, Anže, 2023, Sinteza različnih fluoroforov z azidno ali alkinsko skupino in preučevanje vpliva temperature na intenziteto njihove emisije [na spletu]. Magistrsko delo. [Dostopano 4 maj 2025]. Pridobljeno s: https://repozitorij.uni-lj.si/IzpisGradiva.php?lang=slv&id=153166

Sekundarni jezik

• Jezik: Angleški jezik
• Naslov: Synthesis of various fluorophores with an azide or alkyne group and the study of temperature effect on their emission intensity

Izvleček

Temperature affects all biochemical processes. Adaptation to ambient temperature is a key feature that has enabled the survival and development of living beings. Temperature in the cell and its surroundings is a fundamental and extremely important physical parameter for cellular systems and their functions such as cell division, diffusion, metabolism, differentiation, gene expression and enzyme activity. Observing the effects of drugs, disease, aging and other physiological changes in the cell through temperature changes could be monitored in real time using fluorescence. The phenomenon of fluorescence is the emission of light from a substance, which occurs when a substance is irradiated with light of shorter wavelength, the substance absorbs light, which causes an electron to go into an excited state. Upon returning to the ground state, the compound then emits a photon of a higher wavelength than the light used for irradiation. Compounds that are capable of fluorescence are called fluorophores. The fluorescence intensity of some fluorophores is temperature-dependent and they are a good starting point for the development of probes for measuring temperature at the microscopic level (e. g. in organelles of cells). In this master thesis, we synthesized various fluorophores which fluorescence depends on the ambient temperature. We designed fluorophores from the family of anthracenes, xanthenes and coumarins, which were also successfully synthesized. The aim was to make water-soluble probes based on different basic skeletons in order to obtain probes with different fluorescence response to temperature. We have introduced an alkine or azide group to these fluorophores, which allow for easy “click” chemistry. After successful synthesis, the excitation and emission spectra of the selected probes were measured. On their basis we were able to decide on the appropriate compounds for further investigation. We then measured the emission spectra of selected compounds as a function of temperature and combined the corresponding pairs with a click reaction. For further work, we propose the synthesis of the same basic skeletons with other substituents to reduce lipophilicity, improve stability and water solubility.

• Ključne besede: cell temperature, fluorescence, fluorophore, temperature dependence of fluorescence, click reaction