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Nanocomposite with adenosine wrapped by exosome and application thereof

A nanocomposite and exosome technology, applied in the field of nanomedicine, can solve the problems of short blood half-life, achieve the effects of improving learning and memory, improving exploration ability, and reversing the reduction of slope

Pending Publication Date: 2022-05-27
SOUTH CHINA NORMAL UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, as a small molecule drug, adenosine has a short blood half-life, is easily metabolized, and is difficult to exert its effect through the blood-brain barrier. Therefore, it is very important to modify adenosine to exert its therapeutic effect on ischemic stroke

Method used

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  • Nanocomposite with adenosine wrapped by exosome and application thereof
  • Nanocomposite with adenosine wrapped by exosome and application thereof
  • Nanocomposite with adenosine wrapped by exosome and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0065] Example 1 Preparation of a nanoscale exosome-adenosine complex

[0066] 1. Experimental method

[0067] 1. HT22 cell culture

[0068] Take 50ml of fetal bovine serum and put it in a high-speed centrifuge (ultracentrifuge CP80NX), centrifuge for 120 min at 4°C, with a centrifugal force of 100,000g to obtain fetal bovine serum without exosomes. 10% fetal bovine serum in DMEM high-glucose medium (containing double antibodies, the concentration of penicillin is 0.03mg / ml, the concentration of streptomycin is 0.05mg / ml, pH=7.0-7.2), the thawed HT22 hippocampal cells were recovered in The cells were incubated at 37°C in a cell flask. When the cell density reached 90%, the culture medium was discarded, washed twice with PBS, and then digested with 0.25% trypsin containing EDTA to prepare a single cell suspension, which was inoculated into a petri dish. .

[0069] 2. Extraction of exosomes from HT22 cells

[0070] When the cells are covered by about 70% to 80% of the bottom...

Embodiment 2

[0077] Example 2 Physical and chemical characterization and detection of nanoscale exosome-adenosine complex particles

[0078] 1. Scanning electron microscope detection

[0079] 1. Experimental method

[0080] Take a small amount of a certain concentration Example 1 Prepare the nanoscale exosome-adenosine complex solution in an EP tube, put it on ice, take it to the electron microscope room, use a different pipette to absorb a small amount of liquid, drop one drop on 300 mesh On the copper mesh, take it up to dry naturally to ensure that it is fully dried, and then check the particle morphology with a 120KV electron microscope.

[0081] 2. Experimental results

[0082] The result is as figure 2 As shown in the electron microscope image, the exosome-adenosine complex prepared in Example 1 is spherical in shape, with a diameter close to 100-150 nm, and has an obvious membrane structure.

[0083] 2. Grain testing

[0084] 1. Experimental method

[0085] A small amount of ...

Embodiment 3

[0099] Example 3 The effect of nanoscale exosome-adenosine complex on ischemia mouse model

[0100] The successfully synthesized nanoscale exosome-adenosine complex was subsequently applied to ischemia-reperfusion mice to detect its effects on motor behavior and learning and memory ability of ischemic mice.

[0101] When screening the mouse model of cerebral ischemia, the model was established by clipping bilateral common carotid arteries for 30 minutes. After the clipping for 30 minutes, reperfusion injury was performed, and then the surgical wound was sutured. Penicillin injections were given for 3 days after surgery to prevent infection. Pure adenosine and the nanoscale exosome-adenosine complex prepared in Example 1 were injected into the tail vein for 10 days, respectively, followed by cardiac perfusion, and brain tissue was collected for HE staining to observe the damage of cells in the hippocampal CA1 area to determine whether the model Build success.

[0102] A mouse...

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Abstract

The invention discloses a nano compound with adenosine wrapped by exosome and application of the nano compound, a nano drug delivery system is developed and designed by wrapping a small molecule drug adenosine by using endogenous carrier mouse hippocampus cell exosome, the particle size of the nano drug delivery system is about 150nm, and the nano drug delivery system is stable in form. The traditional Chinese medicine composition can improve cognitive impairment caused by cerebral ischemia, improve the exploration ability of ischemic mice, improve the learning and memory of the mice, and play a neuroprotective role in behavioral expression; and meanwhile, the reduction of fEPSP slope can be reversed. The compound can be used for preparing ischemic stroke postoperative nerve injury repairing medicines.

Description

technical field [0001] The present invention relates to the technical field of nanomedicine, in particular, to a nanocomposite of exosome-encapsulated adenosine and its application. Background technique [0002] Stroke, also known as stroke, is a neurological disease with cerebral ischemia and hemorrhagic injury as the main clinical manifestations. Worldwide, stroke is one of the leading causes of death and permanent disability. Due to its extremely high incidence, stroke has become the primary major disease threatening human health, affecting the lives of millions of people every year. Ischemic stroke is the main type of stroke in clinical practice. About 85% of strokes are ischemic in nature. Ischemic stroke is caused by the formation of blood clots resulting in insufficient blood supply (ischemia) and lack of oxygen and nutrients to the brain material deficiency. Subsequently, a series of biochemical reactions leading to neuronal cell death and neuroinflammation occur....

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K9/51A61K47/46A61K31/7076A61P9/10A61P25/00A61P25/28
CPCA61K9/5176A61K31/7076A61P9/10A61P25/00A61P25/28
Inventor 关燕清刘丽陈祉贝张艺泉李梓晴
Owner SOUTH CHINA NORMAL UNIVERSITY
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