OTX015 (500 nM) exposure induces a strong decrease of BRD2, BRD4 and c-MYC and increase of HEXIM1 proteins, while BRD3 expression is unchanged. c-MYC, BRD2, BRD3, BRD4 and HEXIM1 mRNA levels do correlate however with viability following exposure to OTX015. Sequential combinations of OTX015 with other epigenetic modifying drugs, panobinostat and azacitidine have a synergic effect on growth of the KASUMI cell line[2]. OTX015 (0.1, 1, 5 μM) treatment induces HIV-1 full-length transcripts and viral outgrowth in resting CD4+ T cells from infected individuals receiving suppressive antiretroviral therapy (ART), while exerting minimal toxicity and effects on T cell activation. OTX015-mediated activation of HIV-1 involves an increase in CDK9 occupancy and RNAP II C-terminal domain (CTD) phosphorylation.
In MDA-MB-231 murine xenografts, tumor mass is significantly (p < 0.05) reduced by OTX015 (50 mg/kg) with respect to vehicle-treated animals. OTX015 in combination with 2 mg/kg everolimus shows more effective activity than OTX015 alone.
TZMbl cells, grown in Dulbecco’s modified Eagle’s medium (DMEM) supplemented with 10% FBS, are plated at 1×105 cells/well in 24-well culture plates 24 h before transfection and then transfected with Tat or pcDNA 3.1 plasmid using Lipofectamine 2000. Primary CD4+ T cells (5 × 106) isolated from healthy donors are transfected with Tat or pcDNA 3.1 plasmid at the presence of LTR-Luc construct and then plated in 6-well culture plates. At 24 h post-transfection, the cells are mock-treated or treated with OTX015. At 48 h post-treatment, cells are lysed and luciferase activity is measured using the Dual-Luciferase Reporter Assay Kit.
MCE has not independently confirmed the accuracy of these methods. They are for reference only.
