Signaling pathways, such as the Hedgehog (Hh) pathway, hold the key for unlocking the potential applications of stem cells and are also considered attractive targets for anti-cancer intervention. Although, several small molecule Hh inhibitors are undergoing clinical trials, there is a need to identify new inhibitors capable of overcoming acquired resistance. In this context, a natural product, Taepeenin D, has been recently identified to interfere with Gli function, which is the last step in Hh signaling. The aim of this multidisciplinary project was to develop novel small molecule inhibitors of Gli function based on the structure of Taepeenin D. To this end: i) the first total synthesis of this natural product was pursued and ii) Structure-Activity-Relationships were investigated through biological evaluation of related structural analogues.