Exosome, preparation method thereof, use thereof and pharmaceutical composition

Patent metadata

Field	Value
Publication number	US-2024252542-A1
Application number	US-202418425313-A
Country	US
Kind code	A1
Filing date	Jan 29, 2024
Priority date	Jan 30, 2023
Publication date	Aug 1, 2024
Grant date	—

Abstract

An exosome, a preparation method of an exosome, uses of the exosome, and a pharmaceutical composition including the exosome for treating an ischemia condition of a tissue are provided. The exosome is derived from a genetically engineered mesenchymal stem cell, and the genetically engineered mesenchymal stem cell includes an exogenous PD-L1 gene and an exogenous HGF gene.

First claim

What is claimed is:

1 . An exosome derived from a genetically engineered mesenchymal stem cell, wherein the genetically engineered mesenchymal stem cell comprises an exogenous PD-L1 gene and an exogenous HGF gene.

2 . The exosome of claim 1 , wherein the exogenous PD-L1 gene comprises the amino acid sequence of SEQ ID NO: 4, and the exogenous HGF gene comprises the amino acid sequence of SEQ ID NO: 5.

3 . The exosome of claim 1 , wherein the exogenous PD-L1 gene and the exogenous HGF gene are linked by a self-cleaving peptide coding fragment.

4 . The exosome of claim 1 , wherein the exosome comprises an overexpression PD-L1 and an overexpression HGF.

5 . The exosome of claim 4 , wherein when measured by a flow cytometry, a PD-L1 expression level on an exosome membrane of the exosome is increased relative to a control exosome, and the control exosome is derived from a non-genetically engineered mesenchymal stem cell.

6 . The exosome of claim 4 , wherein when measured by an ELISA, a HGF content of the exosome is increased relative to a control exosome, and the control exosome is derived from a non-genetically engineered mesenchymal stem cell.

7 . The exosome of claim 1 , wherein the exosome comprises an overexpression CXCR4 on an exosome membrane thereof.

8 . The exosome of claim 7 , wherein when measured by a flow cytometry, a proportion of the exosome with CXCR4 surface marker is increased relative to a control exosome, and the control exosome are derived from a non-genetically engineered mesenchymal stem cell.

9 . The exosome of claim 1 , wherein a particle size of the exosome ranges from 30 nm to 200 nm.

10 . The exosome of claim 9 , wherein the particle size of the exosome ranges from 100 nm to 150 nm.

11 . A preparation method of an exosome, comprising:
constructing a genetically engineered mesenchymal stem cell, wherein an exogenous HGF gene and an exogenous PD-L1 gene are transferred into a mesenchymal stem cell to obtain the genetically engineered mesenchymal stem cell; performing a culture step, wherein the genetically engineered mesenchymal stem cell is cultured in a culture medium to obtain a conditioned medium; and performing an isolation step, wherein an exosome is collected from the conditioned medium by an isolating method.

12 . The preparation method of the exosome of claim 11 , wherein in the culture step, the genetically engineered mesenchymal stem cell is cultured under a hypoxic condition.

13 . The preparation method of the exosome of claim 12 , wherein the hypoxic condition is an oxygen content of 3% or less.

14 . The preparation method of the exosome of claim 11 , wherein the mesenchymal stem cell is an adipose mesenchymal stem cell, an umbilical cord mesenchymal stem cell or a bone marrow mesenchymal stem cell.

15 . The preparation method of the exosome of claim 11 , wherein the mesenchymal stem cell is an adipose mesenchymal stem cell or an umbilical cord mesenchymal stem cell.

16 . A pharmaceutical composition for treating an ischemia condition of a tissue, comprising:
the exosome of claim 1 ; and a pharmaceutically acceptable carrier.

17 . The pharmaceutical composition of claim 16 , wherein an administration route of the pharmaceutical composition is intravenous injection, intracarotid artery injection, intraarterial injection or a combination thereof.

18 . A method for treating an ischemia condition of a tissue, comprising administering the exosome of claim 1 to a subject in need for a treatment.

19 . The method of claim 18 , wherein the tissue is a brain.

20 . The method of claim 19 , wherein the ischemia condition is an ischemic stroke.

21 . The method of claim 20 , wherein the exosome reduces an area of the brain damaged by the ischemic stroke in the subject.

22 . A method for enhancing neuroregeneration or reducing neuron death, comprising administering the exosome of claim 1 to a subject in need for a treatment.

23 . A method for reducing an inflammatory response, comprising administering the exosome of claim 1 to a subject in need for a treatment.

Assignees

• Univ China Medical

Inventors

• Jeng Long-Bin
• Hung Mien-Chie
• Shyu Woei-Cherng
• Lin Syuan-Ling
• Chen Chien-Lin

Classifications

• C12N5/06

  Animal cells or tissues; Human cells or tissues (preservation of excised living parts A01N1/10) · CPC title

• A61P25/28

  for treating neurodegenerative disorders of the central nervous system, e.g. nootropic agents, cognition enhancers, drugs for treating Alzheimer's disease or other forms of dementia · CPC title

• A61K35/28 Primary

  Bone marrow; Haematopoietic stem cells; Mesenchymal stem cells of any origin, e.g. adipose-derived stem cells · CPC title

• C12N5/0667

  Adipose-derived stem cells [ADSC]; Adipose stromal stem cells · CPC title

• C12N5/0668

  Mesenchymal stem cells from other natural sources · CPC title