Fragment screening using WAC towards new SMARCA4 inhibitors

Sammanfattning: The drug discovery and development process has changed dramatically during the last decades and the growth of more efficient techniques as high throughput screening combined with combinatorial synthesis using amino acids and carboxylic acids on solid support makes it possible to investigate the affinity of molecules towards a biological target. Fragment based drug discovery (FBDD) has been implemented into different techniques with purpose of discovering new drugs. In this study, a high throughput technique, weak affinity chromatography (WAC) has been used to screen small aromatics ( <400 Da) with purpose of finding a new starting point for a SMARCA4 inhibitor. It is known that the protein is involved in the transcription and repair of DNA, hence the discovery of a new inhibitor is of great interest in medicinal chemistry. To find new inhibitors with high quality data, the need for a more neutral reference column is crucial. It has been observed that the current reference column which consists of ethanolamine have a charge impact on the results, where positively charged molecules being repelled and negatively charged molecules more attracted to the column, consequently the molecules elutes faster and slower respectively. Thus, a reference peptide column was developed by mimicking the isoelectrical point and the amino acid sequence of the protein with the purpose of having a reference column that better mimics the protein. During this study, a total of 216 molecules where synthesized, they were distributed in 13 different libraries and each mix was examined and evaluated after the screenings on WAC. WAC has been proven to be an efficient, sensitive, and a robust method, which also provides the possibility to calculate the dissociation constant (KD). Due to its smoothness, flexibility, and productivity, the technique has very high potential to contribute and to establish more reliable FBDD research.