Zotatifin (eFT226) is a highly effective and sequence-selective inhibitor of the RNA helicase eIF4A. It promotes eIF4A binding to specific mRNA sequences with recognition motifs in the 5′-UTRs, with an IC50 of 2 nM, and disrupts the assembly of the eIF4F initiation complex. By enhancing the interaction between eIF4A and polypurine RNA sequence or structural motifs, zotatifin converts eIF4A into a sequence-specific translation repressor, thereby regulating the translation of certain target genes. It also exhibits anti-tumor activity.