Zingiber officinale has been used as a treatment for osteoporosis in Ayurveda, a traditional Indian medicine since about 2000 years ago. Osmunda japonica has been used as a treatment for rheumatoid arthritis for hundreds of years in Europe, mainly in Spain. Two drug candidates that showed powerful effects in the treatment of osteoporosis in Zingiber officinale and Osmunda japonica were identified through various experiments. The two osteoporosis drug candidates discovered through this study are derivatives with a common benzylideneacetone structure. Benzylideneacetone has a benzylidene ring structure and a combined structure of acetone as the main skeleton. This study intends to describe the process of selecting two of the most promising candidates for osteoporosis treatment through in vitro and in vivo experiments. Similar substances are included in Osmunda japonica and Zingiber officinale, and are being studied in various fields such as immunity, inflammation, and cancer. For these experiments, we used various benzylideneacetone derivatives synthesized in Gyeonggido Business & Science Accelerator.Osteoporosis is mainly caused by two causes, and the first is the increase in osteoclast activity. It is known that there are two cytokines that play an important role in causing this phenomenon. First, macrophage colony-stimulating factor (M-CSF) undergoes osteoclast differentiation through the ERK and PI3K/Akt pathways. Second, the receptor activator of nuclear factor-kB ligand (RANKL) binds to its receptor RANK and it induces osteoclast differentiation by delivering RANK signal to NF-kB, Nuclear Factor Of Activated T-Cells 1 (NFATc1). Second, osteoblast activity is reduced. Osteoblasts are regulated by various hormones, growth factors, and cytokines. In this study, changes in RUNX2 (Runt-related transcription factor 2), an early transcription factor in osteoblast differentiation, were studied, and the extent of bone formation was confirmed through ALP (alkaline phosphatase) analysis.In this situation, the current osteoporosis drug development strategy is focused on reducing bone resorption and loss by inhibiting the differentiation of osteoclasts. Alendronate, a bisphosphonate class, is a representative drug. However, side effects caused by excessive osteoclast suppression are a major obstacle to treatment. In order to solve this problem, previous study conducted on a candidate material for a treatment for bone-related diseases that is safe and effective with fewer side effects while maintaining the balance of osteoclasts and osteoblasts. As a result, Osmundacetone, a benzylideneacetone derivative with bone loss inhibitory activity, was found in the Osmunda japonica rhizome extract, and was defined as compound 1 in this paper. Compound 1 (IC50: 7.8 uM) had a lower ability to inhibit osteoclast formation than alendronate. However, as a result of the study through RUNX2 (Runt-related transcription factor 2) and ALP analysis, the ability to induce osteoblast differentiation was increased by about 3 times.In this paper, various derivatives were synthesized based on the previously discovered compound 1, and they were named Benzylideneacetone derivatives. Based on various studies, Iso-dehydrozingerone with strong osteoclast inhibitory ability could be found and compound 2c was defined. Compound 2c has a very similar structure to Dehydrozingerone (Benzylideneacetone derivative), which can be extracted from ginger, in which only the residues 3 and 4 are reversed. Through the study, i am able to obtain the result that compound 2c exhibited 30-fold (IC50: 0.11uM) more potent osteoclastogenesis inhibitory activity than alendronate while maintaining osteoblast activity. In order to verify the effects of compounds 1 and 2c in vivo, a study was conducted in the ddY mouse animal model in which osteoporosis was induced through ovariectomy. As a result of oral administration for a total of 4 weeks after a recovery period of 1 week after surgery, compound 2c administered 1 mg/kg/d displayed a preventive effect on the development of osteoporosis in OVX mice comparable to that of 10 mg/kg/d alendronate. Based on the results of this study confirming the effectiveness of osteoporosis treatment in various in vitro and in vivo study, i found that compounds 1, 2c could inhibit osteoclast formation and independently activate osteoblast formation. This result will suggest the possibility of a new type of osteoporosis treatment in the treatment market where only one of the existing osteoclasts and osteoblasts can be controlled.These characteristics are the advantages of compound 1, 2c that existing treatments do not have, and are expected to provide clinical implications and insights in the development of osteoporosis treatments.