CMLD-2, an inhibitor of HuR-ARE interaction , competitively binds HuR protein disrupting its interaction with adenine-uridine rich elements (ARE)-containing mRNAs ( K i =350 nM). CMLD-2 induces apoptosis exhibits antitumor activity in different cancer cells as colon, pancreatic, thyroid and lung cancer cell lines. Hu antigen R (HuR) is an RNA binding protein, can regulate target mRNAs stability and translation In Vitro CMLD-2 (1-75 μM; 24-72 h) inhibits thyroid cancer cell viability. CMLD-2 (20-30 μM; 24-48 h) activates caspases and induces apoptotic cell death in H1299 and A549 cells. CMLD-2 (30 μM; 24-48 h) induces G1 cell cycle arrest and mitochondrial perturbation in H1299 and A549 cell. CMLD-2 (30 μM; 24-48 h) reduces expression of HuR and HuR-regulated mRNAs and proteins in H1299 cells. CMLD-2 (35 μM; 72 h) decreases directional migration capability in SW1736, 8505C, BCPAP and K1 cells. CMLD-2 induces a strong decrease of MAD2 mRNA levels in SW1736, 8505C, BCPAP and K1 cells. MCE has not independently confirmed the accuracy of these methods. They are for reference only. Cell Viability AssayCell Line: SW1736, 8505C, BCPAP and K1 cells Concentration: 1, 5, 10, 25, 35, 50, 75 μM Incubation Time: 24, 48, 72 hours Result: Reduced the viability of all the four cell lines when used at 35, 50 and 75 μM concentration and at different time points. Apoptosis AnalysisCell Line: H1299, A549, H1975, HCC827, MRC-9 and CCD16 cells Concentration: 20, 30 μM Incubation Time: 24, 48 hours Result: Marked activated the caspase-9 and -3 in lung tumor cells. Induce the cleavage of PARP in lung tumor cells. Significantly increased the annexin-V-positive staining in lung tumor cells. Cell Cycle AnalysisCell Line: H1299, A549, MRC-9 and CCD16 cells Concentration: 30 μM Incubation Time: 24, 48 hours Result: Induced greater G1 phase cell cycle arrest in H1299 and A549 cells than in MRC-9 and CCD16 cells. Western Blot AnalysisCell Line: H1299, A549, H1975, HCC827, CCD16 and MRC-9 cells Concentration: 20, 30 μM Incubation Time: 24, 48 hours Result: Diminished protein expression of HuR, Bcl-2, Cyclin E and Bcl-XL and increased expression of p27 and BAX in lung tumor cells. Form:Solid IC50& Target:HuR