Looking for breakthrough ideas for innovation challenges? Try Patsnap Eureka!

Microvesicles derived from nucleated, mammalian cells and use thereof

a technology of microvesicles and mammalian cells, applied in the direction of microcapsules, chemical/physical processes, dna/rna fragmentation, etc., can solve the problems of inability to deliver drugs to specific cells or tissues, immune responses within, and inability to use red blood cell-derived vesicles to deliver drugs, etc., to reduce the agony and inconvenience of cancer patients, enhance the effect of therapeutic efficacy and easy accurate diagnosis of cells or tissues

Inactive Publication Date: 2014-02-13
MDIMUNE INC
View PDF1 Cites 9 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Benefits of technology

The present invention is about using microvesicles (tiny spheres) made from mammalian cells to deliver therapeutic and diagnostic substances to target cells or tissues. This can be done selectively and effectively, reducing the risk of harmful side effects for patients during treatment. The microvesicles can be particularly useful in treating cancer, even metastasis, without causing harmful side effects. Furthermore, the microvesicles can also be used for accurate diagnosis of cells or tissues associated with diseases. The substances loaded onto the microvesicles can perform their functions effectively. The use of these microvesicles is limited only to the delivery of substances that are expressed by nucleated, mammalian cells. The preparation method of the microvesicles is simple and can be used in both in-vitro and in-vivo treatment and diagnosis, or experiments.

Problems solved by technology

When vesicles, usually comprised of bacterial cell membranes, are derived from Gram-negative bacteria, they have lipopolysaccharides that may cause various adverse effects including immune responses within the body.
However, the red blood cell-derived vesicles cannot be used to deliver drugs to specific cells or tissues because red blood cells lack an ability to recognize specific cells or tissues.
Further, red blood cells are anucleated, therefore, transformation for the expression of ligands recognizing specific cells or tissues on the surface of red blood cell is not possible.

Method used

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
View more

Image

Smart Image Click on the blue labels to locate them in the text.
Viewing Examples
Smart Image
  • Microvesicles derived from nucleated, mammalian cells and use thereof
  • Microvesicles derived from nucleated, mammalian cells and use thereof
  • Microvesicles derived from nucleated, mammalian cells and use thereof

Examples

Experimental program
Comparison scheme
Effect test

example 1

Preparation of Microvesicles by Extrusion

[0187]FIG. 1 is a scheme showing a process of preparing microvesicles loaded with various substances including targeting materials, therapeutic materials and diagnostic materials from nucleated mammalian cells, whether transformed or not.

[0188]According to the procedure illustrated in the scheme of FIG. 1, microvesicles were prepared from monocytes or macrophages. From among those suggested in FIG. 1, extrusion and a density gradient were selected.

[0189]The monocyte U937 (ATCC No. CRL-1593.2) or the macrophage Raw264.7 (ATCC No. TIB-71) was resuspended at a density of 5×106 cells / ml in 3 mL of PBS (phosphate buffered saline). The cell suspension was passed three times through each of the membrane filters with a pore size of 10 μm, 5 μm and 1 μm, in that order. In a 5 mL untracentrifuge tube were sequentially placed 1 mL of 50% OptiPrep, 1 mL of 5% OptiPrep and 3 mL of the cell suspension effluent from the membrane filters. Ultracentrifugation...

example 2

Preparation of Microvesicles by Sonication

[0190]According to the procedure illustrated in the scheme of FIG. 1, microvesicles were prepared from monocytes or macrophages. From among those suggested in FIG. 1, sonication and a density gradient were selected.

[0191]Monocytes or macrophages were suspended at a density of 2×107 cells / ml in 3 mL of PBS, followed by 30 cycles of sonication with the sonicator (UP 400S, Hielscher) at amplitude 50%, and cycle 0.5 and then with a water bath sonicator for 30 min. In a 5 mL untracentrifuge tube were sequentially placed 1 mL of 50% OptiPrep, 1 mL of 5% OptiPrep and 3 mL of the sonicated cell suspension. Ultracentrifugation at 100,000×g for 2 hours formed a layer of microvesicles between 50% OptiPrep and 5% OptiPrep.

example 3

Analysis of Property of Monocyte-Derived Microvesicles

[0192]The microvesicles generated from monocytes in Example 1 were adsorbed for 3 min to a glow-discharged carbon-coated copper grid. The grid was washed with distilled water and stained for 1 min with uranylacetate before observation under a JEM101 electron microscope (Jeol, Japan). The electron microscope image is shown in FIG. 2.

[0193]As can be seen in the Transmission electron microscope(TEM) image of FIG. 2, the microvesicles constructed from monocytes by extrusion consisted of a lipid bilayer and is generally spherical with a size of 100˜200 nm. The microvesicles generated from monocytes in Example 1 were diluted to a concentration of 5 μg / ml in 1 mL of PBS which was then placed in a cuvette and analyzed for particle sizes using a dynamic light scattering (DLS) particle size analyzer. The results are given in FIG. 3. As can be seen, the microvesicles ranged in size from 200 to 300 nm with a mean size of 250 nm.

[0194]With 0....

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

PUM

PropertyMeasurementUnit
period of timeaaaaaaaaaa
sizeaaaaaaaaaa
sizeaaaaaaaaaa
Login to View More

Abstract

The present invention relates to a microvesicle that is derived from nucleated mammalian cells, which are smaller than the nucleated cells. The microvesicles of the present invention can be used in the delivery of a therapeutic or diagnostic substance to specific tissues or cells, and more particularly, relates to microvesicles derived from monocytes, macrophages, dendritic cells, stem cells or the like, which can be used to deliver specific therapeutic or diagnostic substances for treating and / or diagnosing tissue associated with cancer, diseased blood vessels, inflammation, or the like.

Description

TECHNICAL FIELD[0001]The present invention relates to microvesicles derived from nucleated mammalian cells and the use thereof in the delivery of therapeutic and / or diagnostic substances.BACKGROUND ART[0002]A drug delivery system (DDS) is intended to aid the delivery of medicine to a target site within the body to bring about a therapeutic effect. For example, if a medicine is excreted too fast from the body due to its low absorption or bioavailability rates, a DDS may be used to modify the drug release profile. Medicines with serious adverse effects need to be delivered to target tissues or cells only. Many currently available anticancer agents, for example, exhibit cytotoxicity on normal cells as well as on cancerous cells. The substantial delivery of anticancer agents to cancerous cells or tissues would reduce the agony and inconvenience of cancer patients during treatment.[0003]Since the first use thereof in the 1960s, liposomes have been widely studied for their use in DDS. Adv...

Claims

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

Application Information

Patent Timeline
no application Login to View More
Patent Type & Authority Applications(United States)
IPC IPC(8): A61K9/127
CPCA61K9/1278A61K9/127A61K9/5068A61K45/06A61K35/15A61K35/545A61K2300/00
Inventor GHO, YONG SONGKIM, YOON KEUNJANG, SU CHULKIM, OH YOUNCHOI, DONG-SICYOON, YAE JIN
Owner MDIMUNE INC
Who we serve
  • R&D Engineer
  • R&D Manager
  • IP Professional
Why Patsnap Eureka
  • Industry Leading Data Capabilities
  • Powerful AI technology
  • Patent DNA Extraction
Social media
Patsnap Eureka Blog
Learn More
PatSnap group products

Browse by: Latest US Patents, China's latest patents, Technical Efficacy Thesaurus, Application Domain, Technology Topic, Popular Technical Reports.

© 2024 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap|About US| Contact US: help@patsnap.com