Small molecule inhibitors of the Dishevelled‐CXXC5 interaction are new drug candidates for bone anabolic osteoporosis therapy

Bone anabolic agents promoting bone formation and rebuilding damaged bones would ideally overcome the limitations of anti‐resorptive therapy, the current standard prescription for osteoporosis. However, the currently prescribed parathyroid hormone (PTH)‐based anabolic drugs present limitations and adverse effects including osteosarcoma during long‐term use. Also, the antibody‐based anabolic drugs that are currently being developed present the potential limits in clinical application typical of macromolecule drugs. We previously identified that CXXC5 is a negative feedback regulator of the Wnt/β‐catenin pathway via its interaction with Dishevelled (Dvl) and suggested the Dvl–CXXC5 interaction as a potential target for anabolic therapy of osteoporosis. Here, we screened small‐molecule inhibitors of the Dvl–CXXC5 interaction via a newly established in vitro assay system. The screened compounds were found to activate the Wnt/β‐catenin pathway and enhance osteoblast differentiation in primary osteoblasts. The bone anabolic effects of the compounds were shown using ex vivo ‐cultured calvaria. Nuclear magnetic resonance (NMR) titration analysis confirmed interaction between Dvl PDZ domain and KY‐02061, a representative of the screened compounds. Oral administration of KY‐02327, one of 55 newly synthesized KY‐02061 analogs, successfully rescued bone loss in the ovariectomized (OVX) mouse model. In conclusion, small‐molecule inhibitors of the Dvl–CXXC5 interaction that block negative feedback regulation of Wnt/β‐catenin signaling are potential candidates for the development of bone anabolic anti‐osteoporosis drugs. Synopsis Small‐molecule inhibitors of the Dishevelled (Dvl)–CXXC5 interaction activate Wnt/β‐catenin signaling, promote osteoblast differentiation and new bone formation, and are therefore potential anabolic anti‐osteoporosis drug candidates. Small‐molecule inhibitors of the Dvl–CXXC5 interaction were screened via a newly established in vitro binding assay system. The identified compounds promoted osteoblast differentiation and new bone formation. Interactions between the screened compounds and the Dvl PDZ domain were confirmed by nuclear magnetic resonance (NMR) titration analyses. Oral administration of the selected compound showed bone anabolic effects in an ovariectomized mouse model. Graphical Abstract Small‐molecule inhibitors of the Dishevelled (Dvl)‐CXXC5 interaction activate Wnt/β‐catenin signaling, promote osteoblast differentiation and new bon