Synthesis of affinity chromatography compounds for target identification studiesZore, Matej (Avtor) Anderluh, Marko (Mentor) Boije, Gustav (Komentor) Heart failure is a global public health problem that affects millions of people worldwide. Currently available medical therapy provides only a temporary improvement of the symptoms, yet does not stop the progression of the disease. Consequently, there is an unmet need in the development of new therapies to stop progression or even reverse this pathological state. Lately, cardiac transcription factors have been extensively investigated, since they have an essential role in both physiological and pathological processes, including myocardial hypertrophy. Two such transcription factors are GATA4 and NKX2-5, both having a pivotal role in heart development as well as in cardiac hypertrophy. We synthesized a series of compounds for affinity chromatography, based on a potent inhibitor of GATA4-NKX2-5 transcriptional synergy. Designed derivatives have tri- and pentaethylene glycol linker attached at different positions. In later work, two final compounds were assayed for binding affinity with GATA4 and NKX2-5, in order to study the effect of linker position on the ligand-protein affinity. Additionally, control compound was synthesised to study the non-specific binding of used proteins to compounds, linkers, and matrix. The results of the biological assay on GATA4 confirmed that control compound does not bind GATA4, yet there was a presence of some non-specific protein binding. Additionally, it has been observed that linker position does not alter compound-protein binding affinity. On the other hand, binding of NKX2-5 has proved to be non-specific. The described synthesis can be of great help in the synthesis of similar affinity chromatography compounds. Combined with the results of the biological testing, the synthesized compounds are a good basis for further optimization and research in the area. [M. Zore]20182020-10-21 11:06:06Magistrsko delo/naloga121658sl