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KR-20260062161-A - Composition for preventing or treating Parkinson's disease comprising brain endothelial cell-derived nanovesicles as an active ingredient

KR20260062161AKR 20260062161 AKR20260062161 AKR 20260062161AKR-20260062161-A

Abstract

The present invention relates to a composition for the prevention or treatment of Parkinson's disease comprising nanovesicles derived from brain endothelial cells as an active ingredient, and more specifically, to a composition for the prevention or treatment of Parkinson's disease comprising nanovesicles derived from brain endothelial cells surface-decorated with IL-4 as an active ingredient. The IL-4 surface-decorated brain endothelial cell-derived nanovesicles according to the present invention effectively pass through the blood-brain barrier, increase brain endothelial cell activity, induce proliferation of dopaminergic neurons, inhibit dopaminergic neuronal damage, and induce repolarization of brain microglia from an inflammatory state to an anti-inflammatory state; thus, they have the advantage of being usable as a therapeutic agent useful for the prevention, improvement, and treatment of Parkinson's disease.

Inventors

  • 이원종
  • 조하영

Assignees

  • 인천대학교 산학협력단

Dates

Publication Date
20260507
Application Date
20241025

Claims (17)

  1. A pharmaceutical composition for the prevention or treatment of Parkinson's disease comprising nanovesicles derived from brain endothelial cells as an active ingredient.
  2. A pharmaceutical composition for the prevention or treatment of Parkinson's disease according to claim 1, wherein the brain endothelial cell-derived nanovesicles are surface-decorated with cytokines.
  3. A pharmaceutical composition for the prevention or treatment of Parkinson's disease according to claim 1, wherein the brain endothelial cell-derived nanovesicles have an average diameter of 50 to 200 nm.
  4. A pharmaceutical composition for the prevention or treatment of Parkinson's disease according to claim 1, wherein the brain endothelial cell-derived nanovesicle comprises a transferrin receptor on its surface.
  5. A pharmaceutical composition for the prevention or treatment of Parkinson's disease according to claim 1, wherein the brain endothelial cell-derived nanovesicles target and penetrate the blood-brain barrier.
  6. In claim 1, the brain endothelial cell-derived nanovesicles are (i) Increase in brain endothelial cell activity; (ii) Induction of proliferation of dopaminergic neurons; (iii) inhibition of dopaminergic neuronal damage; and/or (iv) Repolarization of brain microglia from an inflammatory state to an anti-inflammatory state; thereby, a pharmaceutical composition for the prevention or treatment of Parkinson's disease that exerts a preventive or therapeutic effect on Parkinson's disease.
  7. A pharmaceutical composition for the prevention or treatment of Parkinson's disease according to claim 6, wherein the pharmaceutical composition inhibits dopaminergic neuronal damage by increasing the gene expression amount of one or more antioxidant factors selected from the group consisting of Nrf2, HO-1, and NQO-1.
  8. A pharmaceutical composition for the prevention or treatment of Parkinson's disease according to claim 6, wherein the pharmaceutical composition reduces inflammation of brain microglia by reducing the gene expression levels of inflammatory factors TNF-α and/or IL-1β.
  9. A pharmaceutical composition for the prevention or treatment of Parkinson's disease according to claim 6, wherein the pharmaceutical composition reduces inflammation of brain microglia by increasing the gene expression levels of the anti-inflammatory factors CD206 and/or Arg-1.
  10. A pharmaceutical composition for the prevention or treatment of Parkinson's disease according to claim 1, wherein the pharmaceutical composition comprises one or more pharmaceutically acceptable carriers.
  11. Cytokine-surfaced brain endothelial cell-derived nanovesicles.
  12. In claim 11, the above-mentioned cytokine is IL-4, IL-13, IL-10, or TGF-β, brain endothelial cell-derived nanovesicle.
  13. A method for preparing cytokine-surface-decorated brain endothelial cell-derived nanovesicles comprising the following steps: (a) A step of extruding brain endothelial cells with an extruder; (b) a step of recovering nanovesicles by centrifuging the extruded liquid passed through the extruder; and (c) A step of surface decorating the surface of the above cytokine on the surface of the nanovesicle.
  14. In Paragraph 13, The above step (a) is characterized by extruding to sequentially pass through 1000nm, 100nm, and 50nm filters.
  15. In Paragraph 13, The above step (b) is characterized by recovering nanovesicles contained in the supernatant after centrifugation at 10,000 to 15,000 rpm for 10 to 30 minutes.
  16. In claim 13, the above step (c) is characterized by surface decorating the surface of the cytokine on the nanovesicle surface by lipid insertion, biotin-streptavidin binding, amide binding, triazole binding, click chemistry reaction, or enzyme conjugation reaction.
  17. In Paragraph 13, The above step (c) is characterized by mixing DSPE-PEG-IL-4 with nanovesicles and then incubating at a temperature of 30-40°C for 2-10 hours to surface decorate the nanovesicles with IL-4.

Description

Composition for preventing or treating Parkinson's disease comprising brain endothelial cell-derived nanovesicles as an active ingredient The present invention relates to a composition for the prevention or treatment of Parkinson's disease comprising nanovesicles derived from brain endothelial cells as an active ingredient, and more specifically, to a composition for the prevention or treatment of Parkinson's disease comprising nanovesicles derived from brain endothelial cells surface-decorated with IL-4 as an active ingredient. Recently, therapies based on extracellular vesicles (EVs), which are produced within cells such as cells and platelets and secreted into the extracellular space, have emerged as a treatment for Parkinson's disease (Theranostics. 2024; 14(8): 3358-3384) (J Biomed Sci. 2024 Sep 5;31(1):87). Extracellular vesicles contain specific molecules such as proteins, lipids, nucleic acids, and metabolites found in cells. They are characterized by a lipid bilayer surrounding these molecules, which stably protects them while enabling their delivery to other cells. Furthermore, due to their low immunogenicity and cytotoxicity, as well as excellent biocompatibility, they are attracting attention as therapeutic agents capable of overcoming the limitations of existing chemical drugs used to treat Parkinson's disease. However, the amount of extracellular vesicles secreted by cells is limited, and the processes for their isolation and purification require a long time and high cost; additionally, there are areas that need improvement in terms of yield. As an alternative to overcome the above limitations, cell-derived nanovesicles (Nonovesicles, NV) produced using cell extrusion have characteristics similar to extracellular vesicles, but can be obtained with relatively less time and cost and with high yield. Meanwhile, Parkinson's disease is a degenerative brain disease characterized by the degeneration of parts of the brain; it is a chronic and progressive disease that primarily affects motor function. Recently, the incidence rate has been continuously increasing due to population aging, and it is particularly high among the population aged 60 and older. To date, treatments that slow the progression of Parkinson's disease or alleviate symptoms are primarily used, and a cure has not yet been developed. In this context, damage and death of dopaminergic neurons are the pathological causes of Parkinson's disease, and an inflammatory environment caused by the activation of microglia can also induce neuronal damage. However, currently used chemotherapy agents such as dopamine, levodopa, and dopamine agonists have biological limitations, such as being unable to cross the blood-brain barrier (BBB), having short plasma half-lives, and having low bioavailability; they are also accompanied by side effects such as nausea, dyskinesia, drowsiness, and insomnia (Nanoscale Adv. 2022 Nov 3;4(24):5233-5244.) Therefore, there is a need to develop treatments that overcome these biological limitations and have fewer side effects. Figure 1 is a schematic diagram showing the process of manufacturing DSPE-PEG-IL4. Figure 2 shows a schematic diagram illustrating the process of producing brain endothelial cell-derived nanovesicles (BNV) and the size and concentration distribution of nanovesicles according to filtering processes of various sizes. Figure 3 shows the results of confirming the presence of transferrin receptors in brain endothelial cells, brain endothelial cell-derived exosomes (BEVs), and brain endothelial cell-derived nanovesicles (BNVs). Figure 4 shows the results of comparing the amount of IL-4 before and after surface decoration of brain endothelial cell-derived nanovesicles (BNV) using ELISA analysis. Figure 5 shows the results of comparing the degree of repolarization of brain microglia activated by brain microglia-derived exosomes (MEV-IL4), brain endothelial cell-derived exosomes (BEV-IL4), and brain endothelial cell-derived nanovesicles (BNV-IL4), respectively, surface decorated with IL-4. Figure 6 shows the results of comparing the degree of absorption into brain endothelial cells of brain endothelial cell-derived nanovesicles (BNV) and brain microglia-derived nanovesicles (MNV). Figure 7 shows the results of comparing the morphology of brain endothelial cell-derived nanovesicles (BNV) and brain endothelial cell-derived nanovesicles surface-decorated with IL-4 (BNV-IL4) using an electron microscope. Figure 8 shows the results of comparing the sizes of brain endothelial cell-derived nanovesicles (BNV) and brain endothelial cell-derived nanovesicles surface-decorated with IL-4 (BNV-IL4) using NTA. Figure 9 shows the results of comparing the zeta potential and polydispersity of brain endothelial cell-derived nanovesicles (BNV) and brain endothelial cell-derived nanovesicles surface-decorated with IL-4 (BNV-IL4) using DLS. Figure 10 shows the results of analyzing the biocompatibility and storage stability of b