Novel Inhibitor of Keap1-Nrf2 Protein–Protein Interaction Attenuates Osteoclastogenesis In Vitro and Prevents OVX-Induced Bone Loss In Vivo
Open accessAbstract
Keap1 interacts with Nrf2 by assisting in its ubiquitination and subsequent proteolysis. By preventing ROS accumulation during RANKL-induced osteoclastogenesis, Nrf2 activation can prevent the differentiation of osteoclasts. Additionally, inhibiting the Keap1-Nrf2 PPI can be an effective strategy for triggering Nrf2 to regulate oxidative stress. Structure-based virtual screening was performed to discover a potentially novel Keap1-Nrf2 PPI inhibitor wherein KCB-F06 was identified. The inhibitory effects of KCB-F06 on osteoclastogenesis were investigated in vitro through TRAP staining and bone resorption assays. An ovariectomy-induced osteoporosis mouse model was applied to evaluate KCB-F06’s therapeutic effects in vivo. Lastly, the underlying mechanisms were explored using real-time PCR, Western blotting, and co-IP assays. KCB-F06 was discovered as a novel Keap1-Nrf2 PPI inhibitor. As a result, the expression of antioxidants (HO-1 and NQO1) was suppressed, hence reducing ROS accumulation during osteoclastogenesis. Subsequently, this caused the inactivation of RANKL-induced IKB/NF-kB signaling. This eventually led to the downregulation of osteoclast-specific proteins including NFATc1, which is an essential transcription factor for osteoclastogenesis. These results demonstrated that Nrf2 activation in osteoclasts is a valuable tool for osteoclastic bone loss management. In addition, KCB-F06 presents as an alternative candidate for treating osteoclast-related bone diseases and as a novel small molecule that can serve as a model for further Keap1-NRF2 PPI inhibitor development.
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Nrf2 activation protects osteoclast precursor cells from apoptosis induced by excessive oxidative stress by upregulating antioxidant enzymes, thereby maintaining a proper balance in osteoclast function.
“Thirdly, it protects osteoclast precursor cells from apoptosis induced by excessive oxidative stress by upregulating antioxidant enzymes, thereby maintaining a proper balance in osteoclast function57.”
From Intelligence-responsive wettability switch coating on magnesium implants for treating osteoporotic fracture (2026) · cites this paper as [57]
The Discussion (page not provided) says Nrf2 protects cells from ROS-induced damage and that KCB-F06 increased the antioxidant enzymes HO-1 and NQO1 while reducing ROS during osteoclast differentiation. It does not report that Nrf2 prevents apoptosis of osteoclast precursor cells or maintains osteoclast function through precursor survival; its reported effect is inhibition of excessive osteoclastogenesis. (LLM, conf=0.88)
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“…NFATc1 56 , as evidenced by Western blot analysis showing downregulated expression of NFATc1 and CTSK. Thirdly, it protects osteoclast precursor cells from apop-tosisinducedbyexcessiveoxidativestressbyupregulatingantioxidant enzymes, thereby maintaining a proper balance in osteoclast function 57 .”
- Authors
- Chen, Z., Yao, H., Encarnacion, A. M., Jeong, J., Choi, Y., Park, S., Lee, S., & Lee, T.
- Affiliations
- Chonnam National University (KR)
- Department of Oral Biochemistry, Dental Science Research Institute, School of Dentistry, Chonnam National University, Gwangju 61186, Republic of Korea
- Venue
- Antioxidants
- Publisher
- MDPI AG
- Published
- 2024-07-15
- Volume
- 13
- Issue
- 7
- Pages
- 850
- ISSN
- 2076-3921 (ISSN-L)
- PMC
- PMC11273523
- OpenAlex
- W4400640543
- Cited by
- 16
- References
- 38
- Citations by year
- 2026: 10 · 2025: 6
- FWCI
- 2.18
- Type
- journal article
- Language
- en
- Open access
- gold
- doi.org
- Licence
- Locations
- Antioxidants (publishedVersion, cc-by)
- PubMed Central (publishedVersion, cc-by)
- PubMed (publishedVersion)
- DOAJ (DOAJ: Directory of Open Access Journals) (submittedVersion)
- Funders
- National Research Foundation of Korea (NRF) (2021R1A2C2094007)
- Topics
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Bone Metabolism and Diseases
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Genomics, phytochemicals, and oxidative stress
- Life Sciences › Neuroscience › Cellular and Molecular Neuroscience › Nuclear Receptors and Signaling
- Keywords
- Keap1-Nrf2 interaction, protein-protein interaction inhibitors, osteoclastogenesis, ovariectomy-induced bone loss, osteoporosis, RANKL, reactive oxygen species, NF-κB signaling, NFATc1, heme oxygenase-1, NQO1, bone resorption, small molecule inhibitors
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