Ethyl 2-((4-Chlorophenyl)amino)thiazole-4-carboxylate and Derivatives Are Potent Inducers of Oct3/4

Abstract

The octamer-binding transcription factor 4 (Oct3/4) is a master gene in the transcriptional regulatory network of pluripotent cells. Repression of Oct3/4 in embryonic stem cells (ESCs) is associated with cell differentiation and loss of pluripotency, whereas forced overexpression in cooperation with other transcriptional factors, such as Nanog, Sox2, and Lin28, can reprogram somatic cells back into pluripotent cells, termed induced pluripotent stem cells (iPSCs). However, random integration and potential tumorigenic transformation caused by viral transduction limit the clinical application of iPSCs. By performing a cell-based high throughput screening (HTS) campaign, we identified several potential small molecules as inducers of Oct3/4 expression. Here we report a lead structure ethyl 2-((4-chlorophenyl)amino)-thiazole-4-carboxylate, termed O4I2, showing high activity in enforcing Oct3/4 expression. On the basis of chemical expansion, we further identified derivatives having increased activities toward Oct3/4 induction. Thus, O4I2 and its derivatives should provide a new class of small molecules suitable for iPSC generation.

The essential role of TAp73 in bortezomib-induced apoptosis in p53-deficient colorectal cancer cells

“…As we described before 49, 50 , HCT116 cells were seeded in a Ø-12mm cover slip coated with Geltrex (Life Technologies, Germany) at a density of 200,000 cells/well. After 24 hrs the cells were incubated with bortezomib at designed concentrations for 24 hrs, fixed with 4% PFA at RT (room temperature) for 15 min, and blocked with blocking buffer (5% goat serum, 1% BSA and 0.3% Triton X-100 in PBS) for 30 min.…”

Methylisoindigo preferentially kills cancer stem cells by interfering cell metabolism via inhibition of LKB1 and activation of AMPK in PDACs

“…Meisoindigo represses activity of CSC-associated genes and reduces stemness of Jopaca-1 A body of experimental results indicates that genes involved in the regulation of pluripotency, such as Oct3/4, Nanog and Sox2 (Cheng et al, 2015a(Cheng et al, , 2015bLonardo et al, 2011), or abundantly expressed during EMT (Chaffer et al, 2013), such as N-Cadherin and ZEB1, could play important roles in maintaining CSCs. Upon treatment with 20 mM meisoindigo for 24 h, we detected a significant reduction in the expression of CSCassociated genes in Jopaca-1 analyzed by qRT-PCR, showing only 40% residual expression for ZEB1 and 10% for Oct3/4, Sox2 and N-Cadherin (Figure 2A).…”

Methylisoindigo and Its Bromo-Derivatives Are Selective Tyrosine Kinase Inhibitors, Repressing Cellular Stat3 Activity, and Target CD133+ Cancer Stem Cells in PDAC

“…Human primary fibroblasts were isolated and cultivated as described previously with positive ethic permission [ 59 , 60 ].…”

“…As previously reported [ 18 , 59 , 60 ], 2 × 10 5 cells were seeded and treated with compounds for 30 min or 2 h. Cells were lysed in Urea-lysis buffer containing 1 mM EDTA, 0.5% Triton X-100, 5 mM NaF, 6 M Urea, 1 mM Na 3 VO 4 , 10 mg/mL Pepstatin, 100 mM PMSF, and 3 mg/mL Aprotinin in PBS. Protein concentrations were normalized to the smallest value and proteins were resolved on 8% SDS-PAGE and blotted to membrane using BlueFlash Large semi-dry blotter from Serva Electrophoresis (Heidelberg, Germany).…”