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US-12622928-B2 - Nanovesicles from adult stem cells and its use for targeted therapy

US12622928B2US 12622928 B2US12622928 B2US 12622928B2US-12622928-B2

Abstract

Disclosed herein is a nanovesicles from adult stem cells containing iron nanoparticles and its use. The nanovesicle according to the present disclosure provides effects of maximizing the efficacy of treating mesenchymal stem cells by pretreating cells with iron nanoparticles; reconstituting the cells in a nano-sized form to facilitate intravenous injection; and increasing the efficiency of targeting disease areas through magnet induction. In particular, the present disclosure can replace mesenchymal stem cells as a cell therapeutic agent, and it can be applied to various diseases as a novel biopharmaceutical drug because it can increase the function and efficiency of an exosome-based therapeutic agent.

Inventors

  • Byung-Soo Kim
  • Han Young Kim
  • In Bo HAN
  • Hemant Kumar

Assignees

  • SEOUL NATIONAL UNIVERSITY R&DB FOUNDATION
  • SUNGKWANG MEDICAL FOUNDATION

Dates

Publication Date
20260512
Application Date
20190129
Priority Date
20180131

Claims (17)

  1. 1 . An artificial iron-nanovesicle, which is an artificial iron-nanovesicle prepared from an adult stem cell comprising iron nanoparticles therein, wherein the nanovesicle is administered intravenously to a subject, and after the administration, the nanovesicle is targeted through a step of targeting the administered nanovesicle to an organ or tissue of the subject by applying magnetism to the organ or tissue of the subject, wherein iron nanoparticles are contained in the artificial iron-nanovesicle, and have a diameter in the range of 10 nm to 15 nm, and wherein the artificial iron-nanovesicle is prepared by the method comprising: a step of pretreating the adult stem cells by culturing them in the presence of iron nanoparticles; and a step of sequentially extruding the pretreated adult stem cells with a membrane filter having different pore sizes.
  2. 2 . The iron-nanovesicle of claim 1 , wherein 1 μg of the nanovesicle comprises 17 ng of nanoparticles.
  3. 3 . The iron-nanovesicle of claim 1 , wherein the adult stem cell is a mesenchymal stem cell.
  4. 4 . The iron-nanovesicle of claim 1 , wherein the nanovesicle has a diameter in the range of 100 nm to 150 nm.
  5. 5 . A method for preparing a nanovesicle comprising iron nanoparticles therein, wherein the method for preparing the nanovesicle according to claim 1 comprises: a step of providing adult stem cells; a step of pretreating the adult stem cells by culturing them in the presence of iron nanoparticles; and a step of sequentially extruding the pretreated adult stem cells with a membrane filter having at least four different pore sizes with a pore size of 10 μm or less in the order of decreasing pore size.
  6. 6 . The method of claim 5 , wherein in the pretreatment step, 1×10 6 cells are treated with iron nanoparticles at a concentration of 40 μg/mL for 16 hours.
  7. 7 . The method of claim 5 , wherein the extrusion is performed such that the pretreated adult stem cells are sequentially extruded with a membrane filter having a pore size of 10 μm, 5 μm, 1 μm, and 400 nm, and nanovesicles having a pore size of 150 nm are obtained as a result.
  8. 8 . The method of claim 5 , wherein the adult stem cells are mesenchymal stem cells.
  9. 9 . An artificial iron-nanovesicle comprising the iron nanoparticles prepared by a method, comprising: a step of providing adult stem cells; a step of pretreating the adult stem cells by culturing them in the presence of iron nanoparticles; and a step of sequentially extruding the pretreated adult stem cells with a membrane filter having at least four different pore sizes with a pore size of 10 μm or less in the order of decreasing pore size, and wherein iron nanoparticles are contained in the artificial iron-nanovesicle, and have a diameter in the range of 10 nm to 15 nm.
  10. 10 . A pharmaceutical composition for the treatment of a spinal cord injury comprising the artificial iron-nanovesicle of claim 1 .
  11. 11 . A pharmaceutical composition for promoting angiogenesis, comprising the artificial iron-nanovesicle of claim 1 .
  12. 12 . A pharmaceutical composition for anti-inflammation comprising the artificial iron-nanovesicles of claim 1 .
  13. 13 . A pharmaceutical composition for the treatment of stroke or myocardial infarction comprising the artificial iron-nanovesicle of claim 1 .
  14. 14 . The pharmaceutical composition according to claim 10 , wherein the pharmaceutical composition is administered intravenously to a subject, and after the administration, the pharmaceutical composition is administered in such a manner that targeted the administered pharmaceutical composition is targeted to a targeted organ or tissue of the subject by applying magnetism to the organ or tissue of the subject.
  15. 15 . The artificial iron-nanovesicle of claim 9 , wherein in the pretreatment step, 1×10 6 cells are treated with the iron nanoparticles at a concentration of 40 μg/mL for 16 hours.
  16. 16 . The artificial iron-nanovesicle of claim 9 , wherein the extrusion is performed such that the pretreated adult stem cells are sequentially extruded with a membrane filter having a pore size of 10 μm, 5 μm, 1 μm, and 400 nm, and nanovesicles having a pore size of 150 nm are obtained as a result.
  17. 17 . The artificial iron-nanovesicle of claim 9 , wherein the adult stem cells are mesenchymal stem cells.

Description

CROSS-REFERENCE TO RELATED APPLICATION(S) This application claims priority to KR Patent Application No. 10-2018-0011928 filed on Jan. 31, 2018, and KR Patent Application No. 10-2019-0010380 filed on Jan. 28, 2019, the content of which is hereby incorporated by reference herein. FIELD OF THE DISCLOSURE The present disclosure relates to exosomes derived from stem cells and targeted therapy using the same RELATED ART Exosomes are vesicles with a size of 50 nm to 200 nm secreted from cells. Since exosomes have genetic or protein information derived from their parent cells, they can deliver the corresponding information to other cells. As such, the exosomes derived from mesenchymal stem cells have a therapeutic efficacy similar to that of mesenchymal stem cells, and thus, a potential of exosomes as a therapeutic agent in the field of cell-free regenerative medicine has been suggested. Korea Patent Application Publication No. 2018-0003322 (published on Jan. 9, 2018) relates to a composition for promoting angiogenesis, comprising an exosome-mimicking nanovesicle derived from adult stem cells and a method for preparing the same, and a nano-sized vesicle contained in a filtrate of adult stem cells, in which two or more sizes of adult stem cells are sequentially passed from a large filter to a small filter with membrane filters of different sizes. Korea Patent Application Publication No. 10-2017-0010956 (Published on Feb. 2, 2017) relates to a composition for preventing or treating diabetes comprising a stem cell-derived vesicle as an active ingredient. However, the composition had no targeting ability for in vivo organs, and systemically administered exosomes were very limited in use because they did not exert sufficient therapeutic effects. Additionally, due to the nature of exosomes being secreted naturally, a very small amount of exosomes is secreted (1 μg to 4 μg of exosomes are secreted from 106 cells per day based on mesenchymal stem cells), there were limitations in supplying exosomes as a treatment. Additionally, in order to increase target efficiency to an organ in need of treatment, most studies or inventions generally employ a method of direct injection into a disease site, rather than using exosomes as an agent for intravenous injection, but the direction injection has problems in that it is invasive and has poor safety. Therefore, there is a need for the development of a production technology which is capable of synthesizing nanovesicles as a synthetic exosome in a high yield and which has an increased target efficiency to an organ requiring treatment is required. SUMMARY OF THE DISCLOSURE Due to the nature of exosomes being secreted naturally, a very small amount of exosomes is secreted (1 μg to 4 μg of exosomes are secreted from 106 cells per day based on mesenchymal stem cells), there were limitations in supplying exosomes as a treatment. Additionally, most studies or inventions generally employ a method of direct injection into a disease site rather than using exosomes as an agent for intravenous injection, which is invasive and has poor safety. In general, the reason for direct injection is that intravenous injection of exosomes has a low targeting efficiency at the corresponding disease area. Accordingly, the present disclosure synthesizes artificial exosomes (i.e., nanovesicles) in a high yield, and the targeting efficiency also shows an improved effect due to magnet induction by coupling with iron nanoparticles. Exosomes are vesicles with a size of 50 nm to 200 nm secreted from cells. Since exosomes have genetic or protein information derived from their parent cells, they can deliver the corresponding information to other cells. Accordingly, the exosomes derived from mesenchymal stem cells have a therapeutic efficacy similar to that of mesenchymal stem cells, and are thus widely used in various diseases as an alternative therapeutic agent. The present disclosure was developed with the goal of promoting functional recovery of a spinal cord injury by synthesizing artificial exosomes derived from mesenchymal stem cells (i.e., iron-nanovesicles) to improve targeting efficacy and therapeutic effects after intravenous injection. The present disclosure can replace mesenchymal stem cells as a cell therapeutic agent, and it can be applied to various diseases as a novel biopharmaceutical drug because it can increase the function and efficiency of an exosome-based therapeutic agent. An aspect of the present disclosure provides an iron nanovesicle derived from adult stem cells containing iron nanoparticles therein. In an embodiment, the nanovesicle according to the present disclosure is administered to a subject in need of administration of the nanovesicle (e.g., a mammal), particularly considering the characteristics of the nanovesicle according to the present disclosure, is administered intravenously, and after the administration, the nanovesicle is targeted through a step of targeting the administered na