Increased FOXO3 nuclear localization is involved in neuroblastoma chemoresistance and tumor angiogenesis. Accordingly, FOXO3 inhibition is a promising strategy for boosting antitumor immune responses and suppressing FOXO3-mediated therapy resistance in cancer cells. However, no FOXO3 inhibitors are currently available for clinical use. Nevertheless, we have recently identified (4-propoxy)phenylpyrimidinylguanidine as a FOXO3 inhibitor in cancer cells in the low micromolar range. Here, we report the synthesis and structure-activity relationship study of a small library of its derivatives, some of which inhibit FOXO3-induced gene transcription in cancer cells in a submicromolar range and are thus 1 order of magnitude more potent than their parent compound. By NMR and molecular docking, we showed that these compounds differ in their interactions with the DNA-binding domain of FOXO3. These results may provide a foundation for further optimizing (4-propoxy)phenylpyrimidinylguanidine and developing therapeutics for inhibiting the activity of forkhead box (FOX) transcription factors and their interactions with other binding partners.

Department of Organic Chemistry Faculty of Science Charles University Albertov 6 Prague 12843 Czech Republic

Department of Pediatrics 1 Medical University Innsbruck Innrain 66 Innsbruck 6020 Austria

Department of Pediatrics 2 Medical University Innsbruck Innrain 66 Innsbruck 6020 Austria

Department of Physical and Macromolecular Chemistry Faculty of Science Charles University Albertov 6 Prague 12843 Czech Republic

Institute of Physiology of the Czech Academy of Sciences Laboratory of Structural Biology of Signaling Proteins Division BIOCEV Prumyslova 595 Vestec 25250 Czech Republic

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