Our previous study has showed that some fatty acids are inhibitory to osteoclastogenesis in vitro. These fatty acids include the saturated fatty acids palmitic acid and stearic acid, and the mono-unsaturated fatty acid palmitoleic acid. However, their potency is relatively modest, likely due to the rapid metabolism. To improve the potency, we have developed the tetrazole analogue based on the backbone of palmitoleic acid. This analogue has displayed the potency about 10-fold higher than the natural compound in inhibiting osteoclast in vitro. The current study evaluated the analogue’s skeletal effect in vivo. Similar to the natural fatty acids mentioned above, the analogue is not water soluble and solvent-based formulation could be toxic to animals. To make the analogue aqueously soluble, the analogue was complexed with the carrier (2-hydroxypropyl)-β-cyclodextrin (β-CD), which contains a hydrophobic cavity and a hydrophilic surface. CD-1 mice were fed low calcium diet for 5 days before daily injection in the hemi-calvaria with two doses (40 and 100 µg/injection) of the analogue/β-CD complex or the equivalent vehicle for consecutive 5 days. Calcein was injected twice while Alizarin complexone was injected once during the study to label the bone apposition. The results show that the lower dose of the analogue significantly increased the bone formation index in inter label width, bone area, mineralization surface/bone surface, mineral apposition rate and bone formation rate. Meanwhile, the lower dose displayed a trend in reducing the bone resorption index with significant reduction seen in TRAP surface/bone surface. However, the higher dose did not have effect on bone formation and resorption index. This study suggests that the analogue is an anabolic compound on bone.