Cyclolinopeptide F, a cyclic peptide from flaxseed inhibited RANKL-induced osteoclastogenesis via downergulation of RANK expression

  • Toshio Kaneda
  • Yuki Nakajima
  • Sae Koshikawa
  • Alfarius Eko Nugroho
  • Hiroshi MoritaEmail author
Original Paper


Previously, we reported that cyclolinopeptides (CLs) extracted from flaxseed inhibited receptor activator of nuclear factor κ-B ligand (RANKL)-induced osteoclastogenesis from mouse bone marrow cells in vitro. However, mode of action involved in CLs-inhibited osteoclastogenesis has been yet unknown. Therefore, in this study, we investigated the details of inhibitory activity of cyclolinopeptide-F (CL-F) in osteoclastogenesis, as a representative of CLs. CL-F dose-dependently inhibited RANKL-induced osteoclastogenesis (IC50 0.58 µM) without cytotoxic effects. The inhibition by CL-F was mainly observed in macrophage colony-stimulating factor (M-CSF)-induced proliferation/differentiation phase from M-CSF responsive immature myeloid cells to monocyte/macrophage (M/Mϕ) lineage. Additionally, CL-F also slightly inhibited RANKL-induced differentiation phase from M/Mϕ to mature osteoclasts. Expression of RANKL receptor, RANK, in M-CSF-induced M/Mϕ, i.e. osteoclast progenitor cells, was decreased by CL-F treatment. Furthermore, RT-PCR analysis revealed that CL-F inhibited c-fos gene expression, which is reported to be crucial for RANK expression in osteoclast progenitor cells induced with M-CSF from myeloid lineage cells. These results suggested that CL-F inhibits osteoclastogenesis via down regulation of c-fos expression, which leads to the down-regulation of RANK expression in M-CSF-induced osteoclast progenitors.


Flaxseed Cyclic peptide Cyclolinopeptide Osteoclastogenesis Bone marrow macrophage Osteoclast 



This work was partly supported by a Grant-in-Aid for Scientific Research from the Ministry of Education, Culture, Sports, Science and Technology of Japan.

Supplementary material

11418_2019_1292_MOESM1_ESM.pdf (101 kb)
Supplementary material 1 (PDF 100 kb)


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Copyright information

© The Japanese Society of Pharmacognosy 2019

Authors and Affiliations

  • Toshio Kaneda
    • 1
  • Yuki Nakajima
    • 1
  • Sae Koshikawa
    • 1
  • Alfarius Eko Nugroho
    • 1
  • Hiroshi Morita
    • 1
    Email author
  1. 1.Faculty of Pharmaceutical SciencesHoshi UniversityTokyoJapan

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