Absorption enhancers have been investigated since the 1960s, in order to assist the transfer of drugs across the paracellular space in the intestinal epithelium. However, few absorption enhancers are presently used clinically, due to the difficulty of developing enhancers with high specificity and low toxicity. Using high-throughput genomic techniques, new drug candidates such as, non-Lipinski molecules, peptides, antibodies and nucleic acids, are being discovered, so the need for oral drug delivery strategies using absorption enhancers is gaining importance. The key to addressing this issue is to understand the molecular mechanism of the paracellular route in epithelial cell sheets. Towards this end, basic research in cell biology has revealed the components that regulate the paracellular route, and how the transport of substances is regulated. Based on these findings, novel strategies for enhancing drug absorption have been proposed. In this article, the authors first survey the development of absorption enhancers, then outline recent progress in the cell biology of tight junctions, and finally discuss novel approaches for absorption enhancers based on these advances.