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KR-20260067135-A - Composition for Prevention or Treatment of bone diseases comprising comprising Irilin D

KR20260067135AKR 20260067135 AKR20260067135 AKR 20260067135AKR-20260067135-A

Abstract

The present invention relates to a composition for the prevention or treatment of bone diseases comprising irilin D as an active ingredient. Specifically, since irilin D does not cause cytotoxicity and has an excellent effect in inhibiting the differentiation of bone marrow cells into osteoclasts that destroy or absorb bone tissue, it can be usefully utilized as a pharmaceutical composition for the prevention or treatment of bone diseases caused by osteoclast differentiation, such as osteoporosis, osteolithiasis, osteosclerosis, arthritis, Paget's disease, osteoclastoma, periprosthetic osteolysis, cancer bone metastasis, metabolic bone diseases, multiple myelomas, periodontal disease, and osteogenesis imperfecta, or as a health food composition for improvement.

Inventors

  • 이정형
  • 민병선
  • 갈민주

Assignees

  • 강원대학교산학협력단

Dates

Publication Date
20260512
Application Date
20241105

Claims (8)

  1. A composition for the prevention or treatment of bone diseases comprising irilin D as an active ingredient.
  2. In paragraph 1, A composition for the prevention or treatment of bone disease, characterized in that the bone disease is one or more diseases selected from the group consisting of osteoporosis, osteolithiasis, osteosclerosis, arthritis, Paget's disease, osteoclastoma, periprosthetic osteolysis, cancer bone metastasis, metabolic bone disease, multiple myelomas, periodontal disease, and osteogenesis imperfecta.
  3. In paragraph 1, The above composition for the prevention or treatment of bone disease is characterized by preventing or treating bone disease through the inhibition of osteoclast differentiation.
  4. In paragraph 1, The above composition for the prevention or treatment of bone disease is characterized by increasing bone mineral density (BMD), cortical thickness (Ct.Th), femoral cortical area (Ct. Ar/Tt. Ar), trabecular thickness (Tb.Th), number of trabecular teeth (Tb.N), or bone volume ratio (BV/TV), or decreasing trabecular separation (Tb.Sp).
  5. In paragraph 1, The above composition for the prevention or treatment of bone disease is characterized by inhibiting the expression of p-ERK-1/2, p-JNK, p-p38, cathepsin K, TRAP, ACP5, DC-STAMP, c-Fos, and NFATc1 or the degradation of IκBα.
  6. In paragraph 1, The above composition for the prevention or treatment of bone disease is characterized by inhibiting the nuclear translocation of p65/RelA.
  7. A health functional food for the prevention or improvement of bone diseases containing irilin D as an active ingredient.
  8. In paragraph 1, A health functional food for the prevention or improvement of bone diseases, characterized in that the above bone disease is one or more diseases selected from the group consisting of osteoporosis, osteolithiasis, osteosclerosis, arthritis, Paget's disease, metabolic bone disease, multiple myeloma, periodontal disease, and osteogenesis imperfecta.

Description

Composition for Prevention or Treatment of bone diseases comprising Irilin D as an active ingredient The present invention relates to a composition for the prevention or treatment of bone diseases comprising irilin D as an active ingredient. The dynamic equilibrium between osteoclasts (OC), which resorb bone, and osteoblasts (OB), which form bone, is crucial for maintaining bone homeostasis. An imbalance between OCs and OBs can lead to numerous bone disorders, including osteoporosis, osteolithiasis, bone metastasis, and rheumatoid arthritis (Rodan and Martin, 2000; Xu et al., 2023). In particular, the formation or activation of excessive OCs due to chronic inflammation or menopause results in significant bone loss, leading to inflammatory bone diseases and menopausal osteoporosis (Redlich and Smolen, 2012; Mundy, 2007). Therefore, targeting the inhibition of OC differentiation can be the most fundamental approach to preventing and treating osteolytic bone diseases. Osteocysts (OCs), the major multinucleated cells responsible for bone resorption, are distinct from the hematopoietic monocyte/macrophage lineages of the bone marrow (Udagawa et al., 1990). The differentiation of OC precursors into fully functional OCs is driven primarily by macrophage colony-stimulating factor (M-CSF) and nuclear factor-κB ligand receptor activator (RANKL), which are produced by stromal cells, OBs, and osteocytes (Nakashima et al., 2011; Zhou et al., 2022). Furthermore, immune cells in the bone microenvironment secrete cytokines such as RANKL and tumor necrosis factor-α (TNF-α), which can further promote OC differentiation under pathological conditions such as chronic inflammation and estrogen deficiency (Mundy, 2007; Zhou et al., 2022; Veis and O'Brien, 2023). M-CSF not only induces the expression of receptor activators of NF-κB (RANK), the RANKL receptor, but also induces the survival and proliferation of early-stage OC precursors (Kong et al., 1999; Hamilton, 2008; Takayanagi, 2007). The binding of RANKL to RANK of OC precursors initiates the activation of downstream signaling proteins, including extracellular signal-regulating kinase-1/2 (ERK-1/2), c-Jun N-terminal kinase (JNK), p38 MAPK, and the NF-κB pathway (Wada et al., 2006). These signaling pathways activate the c-Fos transcription factor, which consequently leads to the activation of NFATc1 (Boyle et al., 2003; Grigoriadis et al., 1994; Takayanagi et al., 2002). NFATc1 activates the transcription of many OC-specific genes involved in OC differentiation and maturation, including OSCAR, SRC, ITGB3, ACP5, CtsK, and DC-STAMP (Asagiri and Takayanagi, 2007). Bisphosphonates are commonly prescribed as drugs that inhibit the activation of OC. Bisphosphonates prevent bone density loss, thereby reducing the risk of fractures in postmenopausal women with osteoporosis. However, long-term use of such anti-absorption drugs can lead to serious side effects, such as osteonecrosis of the jaw (Skjødt et al., 2018). Therefore, there is a need to develop new absorption inhibitors for the prevention and treatment of OC-related diseases. Figure 1 shows the chemical structure of irilin D (IRD) and its effect on RANKL-induced OC differentiation in mouse BMMs. (A) Chemical structure of irilin D (IRD). (B) Cell viability via CCK-8 analysis after 48 hours of incubation of BMMs with IRD (0, 1, 3, 10 μM) in the presence of M-CSF. (C) TRAP staining images and quantification of TRAP-positive OCs per field after 6 days of incubation of BMMs with IRD (0, 1, 3, 10 μM) in the presence of RANKL and M-CSF. (D) TRAP staining images and quantification of TRAP-positive OCs per field after 6 days of incubation of BMMs with alendronate (AN, 0, 0.3, 1, 3 μM) in the presence of RANKL and M-CSF. (E) Western blotting results of cathepsin K and quantification of α-tubulin-standardized cathepsin K expression levels after culturing RAW264.7 cells with IRD (0, 1, 3, 10 μM) in the presence of RANKL for 3 days. (*P<0.05, **P<0.01, ****P<0.0001. ns: no significant difference) Figure 2 shows the results regarding the effect of irilin D on OC formation in mouse BMMs. (A) Images visualizing the actin structure of OCs by leuorescein-binding phallodin staining and quantification of the number of actin rings per field after incubating BMMs with IRD (0, 1, 3, 10 μM) in the presence of RANKL and M-CSF for 10 days. (B) Representative images of resorption pits in OCs and quantification of the resorption pit area per field after incubating BMMs with IRD (0, 1, 3, 10 μM) in the presence of RANKL and M-CSF for 10 days and performing bone resorption analysis. (*P<0.05, ***P<0.001, ****P<0.0001. ns: no significant difference) Figure 3 shows the results regarding the effect of Irilin D on the differentiation stage of OC. (A) Time-course experimental design. BMMs were cultured with RANKL and M-CSF and then treated with IRD (10 μM) on the indicated dates. (B) Representative image of TRAP staining. (C) Quantification of TRAP-positive OCs