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EP-4577640-B1 - COMPOSITION COMPRISING AVENANTHRAMIDE C FOR INDUCING DIFFERENTIATION OF MESENCHYMAL STEM CELLS INTO CHONDROCYTES AND METHOD OF USING THE SAME

EP4577640B1EP 4577640 B1EP4577640 B1EP 4577640B1EP-4577640-B1

Inventors

  • KHEMARANGSAN, Veerapol

Dates

Publication Date
20260506
Application Date
20231030

Claims (13)

  1. A composition for inducing differentiation of mesenchymal stem cells (MSCs) into chondrocytes, said composition comprising Avenanthramide C (Avn C) and chondroitin sulfate (CS) as a bioactive factor, wherein said composition is a medium or hydrogel.
  2. The composition according to claim 1, wherein said Avenanthramide C (Avn C) is present at the concentration in the range of 10 nM - 50,000 nM, preferably at a concentration of 50 - 300 nM.
  3. The composition according to claim 2, wherein said chondroitin sulfate (CS) is present at the concentration in the range of 300 - 700 µg/mL.
  4. The composition according to claim 1, wherein said mesenchymal stem cells are adipose-derived mesenchymal stem cells (aMSCs).
  5. The composition according to any one of claims 1-4, wherein said chondrocytes are 2-dimensional manner chondrocyte-like cells.
  6. The composition according to any one of claims 1-4, for increasing chondrocyte markers in said chondrocytes, and wherein said chondrocyte markers are selected from the group consisting of FSTL-1, LRP1, ACAN, COL2A, SOX9, and any combination thereof.
  7. Use of a composition according to anyone of claims 1 to 4 for inducing in vitro differentiation of mesenchymal stem cells (MSCs).
  8. A method for inducing differentiation of in vitro cultured mesenchymal stem cells (MSCs) into chondrocytes, said method comprising a step of culturing said MSCs in a medium comprising Avenanthramide C (Avn C) and chondroitin sulfate (CS) as a bioactive factor.
  9. The method according to claim 8, wherein said Avenanthramide C (Avn C) is present at the concentration in the range of 10 nM - 50,000 nM, preferably present at the concentration in the range of 50 nM - 300 nM.
  10. The method according to claim 9, wherein said chondroitin sulfate (CS) is present in the medium at the concentration in the range of 300 - 700 µg/mL.
  11. The method according to claim 8, wherein said mesenchymal stem cells are adipose-derived mesenchymal stem cells (aMSCs).
  12. The method according to any one of claims 8 to 11, wherein said chondrocytes are 2-dimensional manner chondrocyte-like cells.
  13. The method according to any one of claims 8 to 11, wherein chondrocyte markers of said chondrocytes are increased, and wherein said chondrocyte markers are selected from the group consisting of FSTL-1, LRP1, ACAN, COL2A, SOX9, and any combination thereof.

Description

TECHNICAL FIELD The present invention is generally related to an application of Avenanthramide C (Avn C) as an inducer of chondrocyte differentiation in Mesenchymal Stem Cells (MSCs). More particularly, the presently disclosed embodiments are related to the differentiation of mesenchymal stem cells (MSCs) into chondrocytes, a composition comprising avenanthramide c (Avn C), and a method of using the same. BACKGROUND In the context of aging populations, the decrease in operational chondrocytes, identified by their limited regenerative ability, significantly contributes to the gradual worsening of joint discomfort over an extended timeframe. Chondrocytes, essential cells responsible for producing and releasing various components of the extracellular matrix, encompassing diverse types of collagens, are vital for upholding cartilage flexibility. The introduction of chondrocyte replacement therapy has the potential to notably improve the quality of life for individuals grappling with joint-related concerns. Mesenchymal Stem Cells (MSCs), possessing the capacity to differentiate into various cell lineages, including chondrocytes, have generated significant interest in the development of MSC-based therapies to restore cartilage. By utilizing the inherent characteristics of a patient's MSCs and orchestrating their directed differentiation toward the chondrocyte lineage, this inventive approach offers a secure alternative that effectively avoids potential risks linked to immune system rejection of cellular grafts. Mesenchymal Stem Cells (MSCs) are adult stem cells derived from diverse tissues, holding great potential for regenerative cartilage therapies due to their multipotent differentiation abilities. However, a practical and dependable method for inducing chondrocyte differentiation in MSCs is necessary. Current methods lack optimal effectiveness, prompting the requirement for a novel inducer with superior results. Avenanthramides (Avn), polyphenolic compounds predominantly found in oats, have gained attention due to their distinctive biological properties, including antioxidant, antiproliferative, anti-histamine, and anti-inflammatory effects. Korean patent application KR20210091080A1 titled "Pharmaceutical composition for treating or preventing arthritis" and filed by RYU JE HWANG et al. discloses the use of Avn-C to decrease the expression of cartilage matrix-degrading enzyme and cartilage degeneration inducers of chondrocytes for the treatment or prevention of arthritis. Document FU RONG ET AL: 2022 (DOI: 10.1016/j.bcp.2022.115021) relates to the senescence-inducing effect of AVN C in colorectal cancer cells. Further, a non-patent literature titled "Avenanthramide C as a novel candidate to alleviate osteoarthritic pathogenesis" published by Thanh-Tam Tran et al. discloses the use of Avenanthramide-C for the prevention of osteoarthritis (OA) progression and to prevent the expression of matrix-metalloproteinases (MMPs) in OA pathogenesis. This non-patent literature also highlights that the administration of Avn-C did not affect the expression of inflammatory mediators (Ptgs2 and Nos) or anabolic factors (Col2a1, Aggrecan, and Sox9) in OA pathogenesis. Another non-patent literature titled "Avenanthramides Prevent Osteoblast and Osteocyte Apoptosis and Induce Osteoclast Apoptosis in Vitro in an Nrf2-Independent Manner" published by Gretel G. Pellegrini et al. discloses the application of Avenanthramide to enhance osteogenic differentiation by increasing the expression of COL1A. The study demonstrates that the Avenanthramides exert their anti-apoptotic effects on osteoblastic cells and their pro-apoptotic effects on osteoclasts. Another, recent research conducted by Trans et al. sheds light on the inhibitory effects of Avenanthramide C (Avn C) on cartilage degradation. The study revealed that Avn C effectively suppressed the expression of catabolic factors, specifically MMP-3, -12, and -13, induced by IL-1β in mouse articular chondrocytes. However, Avn C did not impact the IL-1β-mediated reduction of mature factors such as Collagen 2 (COL2A1), a major component of the cartilage extracellular matrix (ECM), or SOX9, a crucial transcription factor involved in collagen 2 regulation. Notably, Avn C was applied to inflammatory responses of fully differentiated chondrocytes within mature cartilage during this investigation. This specification recognizes there is a need for a novel application of Avenanthramide C (Avn C) as a potent inducer of chondrocyte differentiation in Mesenchymal Stem Cells (MSCs), establishing a more effective approach in the field. MSCs, which are adult stem cells derived from various tissues such as bone marrow, adipose tissue, and umbilical cord tissue, possess remarkable versatility and can differentiate into multiple cell types. The above-mentioned shortcomings, disadvantages, and problems are addressed herein and which will be understood by reading and studying the following specification. S