Abstract
Background: Osteoporosis affects millions of people worldwide, and current medications such as bisphosphonates and denosumab are not effective enough to reverse bone loss. Moreover, these treatments have drawbacks, including jaw osteonecrosis and skin eczema. Hence, there is an urgent need for new drugs to treat osteoporosis. Methods: Drug library screening was performed via alkaline phosphatase (ALP) staining in osteoblasts to identify potential candidates for osteoporosis treatment. qPCR, Western blotting, ALP staining, alizarin red staining, and tartrate-resistant acid phosphatase (TRAP) staining were conducted to assess the impact of ZM-306416 (ZM) on osteoblast and osteoclast differentiation in vitro. Additionally, RNA sequencing and pathway analysis were carried out to explore the underlying molecular mechanisms involved. Micro-CT scanning and immunostaining were used to determine bone phenotypes in vivo. Results: Drug library screening revealed that ZM enhances ALP activity in osteoblasts, indicating its potential as a pro-osteogenic agent. ZM exerts dual effects by promoting osteoblast differentiation through the Wnt/β-catenin signaling pathway and simultaneously inhibiting osteoclast differentiation through the NF-κB and MAPK signaling pathways. In an OVX mouse model, ZM effectively prevents bone loss by stimulating osteoblast formation and inhibiting osteoclast development. Conclusions: Our study revealed that ZM has a dual anti-osteoporosis effect by promoting osteoblastogenesis and inhibiting osteoclastogenesis, which is mediated by activation of the Wnt/β-catenin signaling pathway and suppression of the NF-κB/MAPK cascades. These findings suggest that ZM could be a promising therapeutic agent for alleviating osteoporosis.