CN-120437124-B - Application of small molecule ISX-9 in promoting mesenchymal stem cell osteogenic differentiation and bone injury regeneration and repair

Abstract

The invention provides application of isoxazole 9 (ISX-9) in promoting mesenchymal stem cell osteogenic differentiation and bone injury regeneration and repair. ISX-9 is a small molecular compound and has the characteristics of easy acquisition, controllable structure, low cost and the like. In-vitro experiments show that ISX-9 has the capability of inducing mesenchymal stem cells to osteogenic differentiation and promoting mineralized nodule formation, and in-vivo experiments show that hydrogel wrapping ISX-9 can promote regeneration and repair of the skull defects of mice, so that ISX-9 has good osteogenesis inducibility and is a potential bone regeneration and repair promoting medicament. Therefore, ISX-9 can promote mesenchymal stem cells to osteogenic differentiation and be applied to bone tissue regeneration and repair. The invention also provides a direct acting target and an acting mechanism of the ISX-9 for promoting osteogenic differentiation. The invention further provides a novel ISX-9 drug sustained release delivery system, which is beneficial to the clinical application from theoretical research.

Inventors

• CUI ZHONGKAI
• CHEN SIXU
• ZHANG JIAN

Assignees

• 南方医科大学

Dates

Publication Date

20260508

Application Date

20250610

Claims (8)

1. The application of ISX-9 in preparing medicines for promoting mesenchymal stem cell osteogenic differentiation is characterized in that the chemical structural formula of ISX-9 is shown as follows: The mesenchymal stem cells are selected from one or more of umbilical cord mesenchymal stem cells, bone marrow mesenchymal stem cells and adipose mesenchymal stem cells, and the osteogenic differentiation is used for bone defect, bone nonunion or delayed repair of bone healing.
2. 2. The application of the hydrogel wrapping ISX-9 in preparing medicines for promoting mesenchymal stem cell osteogenic differentiation is characterized in that the chemical structural formula of the ISX-9 is shown as follows: The mesenchymal stem cells are selected from one or more of umbilical cord mesenchymal stem cells, bone marrow mesenchymal stem cells and adipose mesenchymal stem cells, and the osteogenic differentiation is used for bone defect, bone nonunion or delayed repair of bone healing.
3. 3. The method for inducing the osteoblast differentiation of the mesenchymal stem cells in vitro is characterized in that an osteoblast differentiation medium containing ISX-9 is adopted to carry out the osteoblast differentiation of the mesenchymal stem cells, the ISX-9 is added as an accelerator for inducing the osteoblast differentiation, the concentration of the ISX-9 is 15-25 mu M, the induction time is 7-14 days, and the chemical structural formula of the ISX-9 is as follows: The mesenchymal stem cells are selected from one or more of umbilical cord mesenchymal stem cells, bone marrow mesenchymal stem cells and adipose mesenchymal stem cells.
4. The application of ISX-9 in preparing medicines for repairing bone injury regeneration is characterized in that the bone injury is one or more of bone defect, bone nonunion or bone healing delay, and the chemical structural formula of the ISX-9 is as follows: 。
5. 5. the application of the hydrogel wrapping the ISX-9 in preparing a bone injury regeneration repair drug is characterized in that the chemical structural formula of the ISX-9 is shown as follows: The bone injury is one or more of a bone defect, a bone disjunction, or a delayed bone healing.
6. 6. The use according to claim 2 or 5, wherein the hydrogel is selected from one or more of a glycol chitosan methacrylate hydrogel, a gelatin hydrogel or a hyaluronic acid hydrogel.
7. Use of an ISX-9 drug delivery system for the preparation of a bone injury regeneration repair drug, said ISX-9 having the chemical structural formula: The bone injury is one or more of a bone defect, a bone disjunction, or a delayed bone healing.
8. 8. The use of claim 7, wherein the drug sustained release delivery system comprises one or more of microspheres, micelles, or liposomes.

Description

Application of small molecule ISX-9 in promoting mesenchymal stem cell osteogenic differentiation and bone injury regeneration and repair Technical Field The invention relates to the technical field of stem cells, in particular to application of ISX-9 in promoting mesenchymal stem cell osteogenic differentiation and bone injury regeneration and repair. Background In recent years, mesenchymal stem cells (MESENCHYMAL STEM CELLS, MSCs) have received increasing attention as a model for studying bone tissue regeneration and repair. Stem cells are multipotent cells that have self-renewing ability in vivo and are capable of differentiating into a variety of cell types including osteoblasts under appropriate conditions. Mesenchymal Stem Cells (BMSCs) derived from bone marrow are the most widely studied cells in bone tissue engineering, but acquisition and in vitro culture of BMSCs are a relatively troublesome problem in the clinical application process. Adipose-derived mesenchymal stem cells (Adipose DERIVED STEM CELLS, ADSCS) and BMSCs are derived from mesoderm, have good osteogenic differentiation capacity as BMSCs, and ADSCs have the advantages of wide sources, easier acquisition and small damage to donor areas. ADSCs are preferential candidates for future bone tissue engineering seed cells, and have wide scientific significance and application prospect. Therefore, ADSCs are selected as a cell model research object, and the invention can be applied to mesenchymal stem cells from all mesoderm sources and induced pluripotent stem cells (iPS cells) artificially. The small molecular compound has the unique advantages of targeting, regulating and controlling stem cell fate, controllable cost, easy large-scale preparation and the like, and becomes a research front in the field of bone regeneration. The development of a high-efficiency, safe and economic micromolecular bone repair drug is an important method for solving the clinical dilemma of bone regeneration. ISX-9 is composed of isoxazole and benzene ring, has molecular weight 234.27 Da, and can rapidly penetrate cell membrane to play a role by virtue of the characteristics of small molecular weight and high fat solubility. Recent studies have shown that ISX-9 exhibits significant biological activity in a variety of cell types, particularly with great potential in stem cell fate regulation, cell survival and proliferation. ISX-9 not only can promote calcium ion inflow through activating voltage-gated calcium ion channels and N-methyl-D-aspartate (NMDA) receptors to induce neural stem cells to differentiate towards neurons, but also can penetrate through the blood brain barrier and depend on activation of a muscle cell enhancer 2 (Mef 2), so that proliferation of neural stem cells in hippocampus and neurogenesis （Luis E B Bettio, et al. ISX-9 can potentiate cell proliferation and neuronal commitment in the rat dentate gyrus[J]. Neuroscience, 2016, 332:212-222.）. are promoted, and in addition, the determination of fate of cells from the mesoderm of ISX-9 plays a key regulatory role. ISX-9 activates classical and non-classical WNT signaling pathways at different stages of myocardial progenitor (CPCs) differentiation, promoting myocardial lineage differentiation. ISX-9 can also determine the differentiation direction of enteroendocrine cells (EECs) through the regulation of calcium signals, and determine the lineage fate （Tsakmaki A, et al. ISX-9 manipulates endocrine progenitor fate revealing conserved intestinal lineages in mouse and human organoids[J]. Molecular Metabolism, 2020, 34.）. of intestinal cells although ISX-9 plays an important role in regulating neurogenesis and neuronal function, its role in mesenchymal stem cell osteogenic differentiation and bone injury regeneration repair has not been reported. Disclosure of Invention The invention aims to overcome the defects and the shortcomings in the prior art and provide the application of ISX-9 in promoting mesenchymal stem cell osteogenic differentiation and bone injury regeneration and repair. The research of the invention shows that ISX-9 has the capability of inducing mesenchymal stem cells to osteogenic differentiation and promoting mineralized nodule formation, and in vivo experiments show that hydrogel wrapping ISX-9 can promote regeneration and repair of the skull defect of mice, and the ISX-9 has good osteogenesis inducibility and is a potential bone regeneration and repair promoting medicament. And we further find the mechanism and action target point of ISX-9 for promoting osteogenic differentiation, and construct ISX-9 drug slow release system at the same time, break through the existing hydrophobic micromolecular drug local delivery inefficiency, short technological bottleneck such as acting time, offer brand new target point and intelligent administration strategy for bone injury clinical treatment. The above object of the present invention is achieved by the following technical solutions: The inv