Synthesis and evaluation of 2-NMPA derivatives as potential agents for prevention of osteoporosis in vitro and in vivo

Zhihao Chen; Devaneyan Joseph; Mina Ding; Swapnil Pandurang Bhujbal; Robin Prakash Sirvin Rajan; Eunae Kim; Sang-Wook Park; Sunwoo Lee; Tae-Hoon Lee

doi:10.1016/j.ejmech.2023.115767

Synthesis and evaluation of 2-NMPA derivatives as potential agents for prevention of osteoporosis in vitro and in vivo

Eur J Med Chem. 2023 Nov 15:260:115767. doi: 10.1016/j.ejmech.2023.115767. Epub 2023 Aug 28.

Authors

Zhihao Chen¹, Devaneyan Joseph², Mina Ding¹, Swapnil Pandurang Bhujbal³, Robin Prakash Sirvin Rajan², Eunae Kim⁴, Sang-Wook Park⁵, Sunwoo Lee⁶, Tae-Hoon Lee⁷

Affiliations

¹ BioMedical Sciences Graduate Program (BMSGP), Chonnam National University, Gwangju, 61186, Republic of Korea.
² Department of Chemistry, Chonnam National University, Gwangju, 61186, Republic of Korea.
³ Institute of Pharmaceutical Science and Technology, Hanyang University, Ansan, 426-791, Republic of Korea.
⁴ Department of Pharmacy, College of Pharmacy, Chosun University, Gwangju, 61452, Republic of Korea.
⁵ Department of Oral Biochemistry, Dental Science Research Institute, School of Dentistry, Chonnam National University, Gwangju, 61186, Republic of Korea.
⁶ Department of Chemistry, Chonnam National University, Gwangju, 61186, Republic of Korea. Electronic address: sunwoo@chonnam.ac.kr.
⁷ Department of Oral Biochemistry, Dental Science Research Institute, School of Dentistry, Chonnam National University, Gwangju, 61186, Republic of Korea. Electronic address: thlee83@jnu.ac.kr.

PMID: 37651877
DOI: 10.1016/j.ejmech.2023.115767

Abstract

Abnormal osteoclast differentiation causes various bone disorders such as osteoporosis. Targeting the formation and activation of osteoclasts has been recognized as an effective approach for preventing osteoporosis. Herein, we synthesized eleven 2-NMPA derivatives which are (2-(2-chlorophenoxy)-N-(4-alkoxy-2-morpholinophenyl) acetamides, and evaluated their suppression effects on osteoclastogenesis in vitro by using TRAP-staining assay. Among the synthesized eleven novel 2-NMPAs, 4-(2-(2-chlorophenoxy)acetamido)-3-morpholinophenyl trifluoromethanesulfonate (11b), 4-(2-(2-chlorophenoxy) acetamido)-3-morpholinophenyl-3-(N-(2-oxo-2-((2-(phenylthio) phenyl) amino) ethyl)methylsulfonamido)benzoate (11d), and 4-(2-(2-chlorophenoxy) acetamido)-3-morpholinophenyl 4-acetamidobenzenesulfonate (11h) displayed highly inhibitory bioactivity on the differentiation of primary osteoclasts. 11h was selected for further investigation of the inhibitory effects and potential mechanism involved in the suppression of osteoclastogenesis. In vitro analysis suggested that 11h inhibited osteoclastogenesis with an IC₅₀ of 358.29 nM, decreased the formation of F-action belts and bone resorption, without interfering cell viability and osteoblast differentiation. Furthermore, the mRNA expressions of osteoclast-specific genes such as Acp5, Nfatc1, Dc-stamp, Atp6v0d2, Mmp9, and Ctsk significantly decreased following 11h treatment. RANKL-induced osteoclast-specific proteins analysis demonstrated that 11h suppressed osteoclast differentiation by downregulating of RANKL-mediated TRAF6 expression, followed by inactivation of PI3K/AKT and IκBα/NF-κB signaling pathways. Finally, 11h inhibited ovariectomy-induced bone loss in vivo. Therefore, the current work highlighted the therapeutic potential of 11h as an anti-osteoporosis lead compound.

Keywords: Molecular docking; N-(2-morpholinophenyl) acetamide; Osteoclastogenesis; Structure modification; TRAF6.

MeSH terms

Female
Humans
Osteoclasts
Osteogenesis
Osteoporosis* / drug therapy
Osteoporosis* / prevention & control
Phosphatidylinositol 3-Kinases*

Substances

Phosphatidylinositol 3-Kinases