Target validation of UK-101 and functional studies of beta1i

摘要: OF DISSERTATION TARGET VALIDATION UK-101 AND FUNCTIONAL STUDIES β1i is a major catalytic subunit of the immunoproteasome, an alternative form constitutive proteasome, and its upregulation has been demonstrated in variety disease states including cancer. Our lab developed small molecule inhibitor β1i, dubbed UK-101. While causes apoptosis cancer cell lines, it was not clear whether this apoptotic effect directly mediated by irreversible inhibition β1i. Since off-target effects are roadblocks for development new effective pharmaceuticals, target validation studies system would assist further progression inhibitors towards preclinical trials. hypothesis that expression activity important growth proliferation PC-3 prostate line, therefore seen upon treatment cells with due solely to covalent To test hypothesis, number complementary approaches were used. The increased these interferongamma or tumor necrosis factor-alpha, natural inducers immunoproteasome. decreased using interfering RNA short hairpin RNA, transient stable manner, respectively. All then treated efficacy interferon-gamma but did change any other suggesting specific This confirmed molecular probe proteasome bound subunits. Additional experiments performed determine on cells. Simply removing reduces viability Other mutation which inhibited reduced when compared those containing wild type protein. Overall, our data indicate potential therapeutic Further medicinal chemistry efforts will be required develop into truly selective inhibitor.