Razvoj analiznih metod visoke zmogljivosti za določanje topnosti in porazdelitvenega koeficienta snovi

Žižek, Petra (author), Frlan, Rok (mentor), Proj, Matic (comentor)

Problem lažno pozitivnih zadetkov in zadetkov z neugodnimi lastnostmi za razvoj do spojin vodnic, ki jih dobimo pri rešetanju visoke zmogljivosti, lahko omejimo z dobro pripravo knjižnic spojin. Kemijske knjižnice spojin lahko sestavlja več milijonov spojin, ki imajo želene lastnosti za nadaljnje testiranje. Vsaki kemikaliji lahko določimo lastnosti, kot so kemijska struktura, čistost, količina in fizikalno-kemijske lastnosti spojine in jih shranimo v nekakšni zbirki podatkov. Našo knjižnico, ki vsebuje 887 spojin, smo pripravili s pomočjo virtualnega filtriranja v računalniškem programu KNIME iz kemikalij, ki jih že imamo na naši fakulteti. Cilj magistrske naloge je bil razvoj in optimizacija dveh metod visoke zmogljivosti za določanje topnosti in porazdelitvenega koeficienta, ki bi ju lahko uporabili na pripravljeni kemijski knjižnici. Za določanje topnosti smo uporabili metodo merjenja absorbance na mikrotitrski plošči in metodo HPLC. Na mikrotitrskih ploščah smo določali kinetično in termodinamsko topnost, ter preverili termodinamsko metodo še s pomočjo HPLC metode na reverznofazni C18 koloni. Za določanje porazdelitvenega koeficienta smo uporabili izokratsko metodo HPLC. Preizkušali smo tri različne kolone, različne mobilne faze in deleže organskega modifikatorja. V drugem delu smo metodi uporabili za določanje topnosti in porazdelitvenega koeficienta 219 modelnim spojinam iz kemijske knjižnice, ki smo jo pripravili z izbranimi filtri v računalniškem programu KNIME. Metodo merjenja termodinamske topnosti smo sicer razvili, a žal ni dala dobrih rezultatov, saj so le ti slabo ponovljivi in imajo slabo korelacijo z rezultati HPLC metode. Pravtako nismo bili uspešni z metodo za določanje porazdelitvenega koeficienta, saj le ta ni primerna za uporabo na takšnem številu spojin. Za nadaljnje delo predlagamo še dodaten razvoj in optimizacijo HPLC metode za določanje logP.

• Language: Slovenian
• Keywords: knjižnica spojin, topnost, logP, HPLC, visoka zmogljivost
• Work type: Master's thesis/paper
• Organization: FFA - Faculty of Pharmacy
• Year: 2020
• PID: 20.500.12556/RUL-121726
• Publication date in RUL: 24.10.2020

ŽIŽEK, Petra, 2020, Razvoj analiznih metod visoke zmogljivosti za določanje topnosti in porazdelitvenega koeficienta snovi [online]. Master’s thesis. [Accessed 11 April 2025]. Retrieved from: https://repozitorij.uni-lj.si/IzpisGradiva.php?lang=eng&id=121726

Secondary language

• Language: English
• Title: Development of high-throughput analytical methods for the determination of substances solubility and partition coefficient

Abstract

The problem of false positives and hits with unfavourable properties obtained in high-throughput screening can be limited by good preparation of compound libraries. Chemical libraries can consist of up to a few million compounds that have the desired properties for further testing. Each compound can be characterized by properties such as chemical structure, purity, quantity and physicochemical properties of the compound. Our library, which contains 887 compounds, was prepared using virtual filtering in KNIME software from the chemicals on our faculty. The master's thesis aims to develop and optimize high-throughput methods for determining the solubility and partition coefficient that could be applied to our prepared chemical library. The microplate absorbance measurement method and the HPLC method were used to determine the solubility. We determined kinetic and thermodynamic solubility on microplates and then validated the thermodynamic method using the HPLC method on a reversed-phase C18 column. We used the isocratic HPLC method to determine the partition coefficient. We tested three different columns, different mobile phases and organic modifier proportions. Furthermore, these methods were used to determine the solubility and partition coefficient of 219 model compounds from the chemical library, which was prepared with selected filters in the KNIME software. The method of measuring thermodynamic solubility was developed but unfortunately it did not give good results, as these are poorly reproducible and had a poor correlation with the results of HPLC methods. The method for determining the partition coefficient was not suitable for use on such scale. The Master’s thesis outcomes represent starting points for further development and optimization of the logP HPLC method.

• Keywords: chemical library, solubility, logP, HPLC, high throughput