Določanje termodinamske topnosti in permeabilnostnega koeficienta zaviralcev bakterijskih giraz B in topoizomeraz IV

Nizka topnost je eden izmed najpogostejših vzrokov za prekinitev razvoja učinkovine, zato je topnost treba spremljati že v začetnih fazah razvoja zdravila. Slaba topnost pomeni slabšo absorpcijo spojine, kar se kaže v slabšem učinku zdravila. Ob topnosti v začetnih fazah razvoja učinkovine določamo še permeabilnostne lastnosti, od katerih je odvisno, kako dobro bo spojina prehajala biološke bariere, kot so celične membrane. Permeabilnostne lastnosti nam povedo, kakšna bo pot učinkovine do mesta delovanja in tako pomagajo napovedati ADME lastnosti spojin. Spojinam lahko določimo kinetično ali termodinamsko topnost. Kinetično topnost običajno določamo v zgodnjih fazah raziskovanja učinkovin, termodinamsko pa nekoliko kasneje v fazi razvoja zdravil. V zadnjih letih je prišlo do razvoja številnih metod za določanje topnosti, za referenčne metode pa štejemo različne izvedbe določitve ravnotežne topnosti. Določanje permeabilnostnih lastnosti lahko izvedemo s PAMPA in vitro metodo visoke zmogljivosti, ki uporablja nebiološko membrano in posnema lastnosti bioloških barier. Na katedri za farmacevtsko kemijo na Fakulteti za farmacijo so sintetizirali modelne spojine karboksilne kisline in strukturno različne CF3-substituirane diole, ki predstavljajo ključne fragmente zaviralcev bakterijskih topoizomeraz. V magistrski nalogi smo določili in vrednotili topnost in permeabilnost teh spojin glede na njihove strukturne lastnosti, kjer smo si pomagali s podatki iz literature. Za določanje koncentracij smo uporabili HPLC tehniko, ki je glede na število testiranih spojin dovolj zmogljiva, hkrati pa tudi zadostno občutljiva in selektivna, da z njo lahko kvantitativno določimo vse topnosti in prepoznamo morebitne težave s stabilnostjo preiskovanih spojin med meritvami. Dobljeni rezultati bodo služili kot pomoč pri optimizaciji topnosti in permeabilnosti novih zaviralcev DNA giraze in topoizomeraze IV. Low solubility is one of the most common reasons for interrupting the development of active pharmaceutical ingredients, so it is necessary to determine these properties already in the initial stages. Poor solubility means a poorly absorbed compound, which is reflected in a lower effect of the drug. Along with solubility in the initial stages of development we also determine permeability properties, which determine how well the compound will pass through biological barriers, such as cell membranes. Permeability properties tell us what the path of the active ingredient will be to the site of action and thus help us predict the ADME properties of the compound. For compounds we can determine kinetic or thermodynamic solubility. Kinetic solubility is usually determined in the early stages of active ingredient research and thermodynamic solubility is determined later in the drug development phase. In recent years, a number of methods for determining solubility have been developed, and reference methods include the most diverse implementations of determining equilibrium solubility. Determination of permeability properties can be performed with the high-throughput PAMPA in vitro method, which uses a non-biological membrane and mimics the properties of biological barriers. Model carboxylic acid compounds and structurally different CF3-substituted diols, which represent key fragments of bacterial topoisomerase inhibitors, were synthesized at the Department of Pharmaceutical Chemistry at the Faculty of Pharmacy. In the master's thesis, we determined and evaluated the solubility and permeability of these compounds according to their structural properties, where we used data from the literature. To determine the concentrations, we used the HPLC technique, which, given the number of tested compounds, is powerful enough, but also sensitive and selective enough to quantitatively determine all solubilities and identify potential problems with the stability of the investigated compounds during measurements. The obtained results will serve as an aid in optimizing the solubility and permeability of new inhibitors of DNA gyrase and topoisomerase IV.