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Application of chitosan nanogel loaded with oxytocin in preparation of medicine for early intervention of Alzheimer's disease

A kind of Alzheimer's disease and chitosan nanotechnology, which is applied in the field of biomedicine, can solve the problems of low effective concentration and short half-life of exogenous oxytocin, so as to improve learning and memory ability, reduce Aβ deposition and neurotoxicity. The effect of non-damage and mild reaction conditions

Pending Publication Date: 2021-11-26
TIANJIN MEDICAL UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0006] The purpose of the present invention is to solve the problem that the effective concentration in the brain is low due to the short half-life of exogenous oxytocin, which limits its application to the early intervention of Alzheimer's disease, and provides a chitosan nanometer loaded with oxytocin Application of the gel in the preparation of early intervention drugs for Alzheimer's disease, that is, early intervention for Alzheimer's disease through anti-inflammatory effects

Method used

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  • Application of chitosan nanogel loaded with oxytocin in preparation of medicine for early intervention of Alzheimer's disease
  • Application of chitosan nanogel loaded with oxytocin in preparation of medicine for early intervention of Alzheimer's disease
  • Application of chitosan nanogel loaded with oxytocin in preparation of medicine for early intervention of Alzheimer's disease

Examples

Experimental program
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Effect test

Embodiment 1

[0041]In this example, a non-functional modified chitosan nanogel loaded with oxytocin was prepared. Its preparation method comprises the following steps:

[0042] (1) 10 mg of polyethylene glycol-modified carboxymethyl chitosan was dissolved in 10 mL of ultrapure water, and the pH was adjusted to 5.0.

[0043] (2) 5 mg of oxytocin was added under magnetic stirring (850 rpm), followed by 3.96 mL of TPP solution with a concentration of 0.6 mg / mL, and stirred for 3 minutes.

[0044] (3) The above solution was centrifuged at 9000 rpm for 20 minutes to obtain a non-functionalized modified chitosan nanogel loaded with oxytocin, which was stored at 4° C. in the dark for future use.

[0045] The prepared oxytocin-loaded chitosan nanogel was characterized for its particle size and potential. The results showed that the dynamic light scattering particle size was 91nm, and the surface potential was about 0mV.

Embodiment 2

[0047] In this example, an oxytocin-loaded chitosan nanogel modified with functional molecules across the blood-brain barrier was prepared. Its preparation method comprises the following steps:

[0048] (1) Dissolve 10 mg of polyethylene glycol-modified carboxymethyl chitosan in 10 mL of ultrapure water, and then add 400 μL of 1-(3-dimethylaminopropyl)-3-ethyl carbon with a concentration of 10 mg / mL Diimine hydrochloride and 200 μL of N-hydroxysuccinimide at a concentration of 10 mg / mL were magnetically stirred in ultrapure water for 1 hour.

[0049] (2) 3.5 mg of angiopep-2 was added to the above solution, magnetically stirred for 6 hours under dark conditions, and then dialyzed in ultrapure water for 24 hours under dark conditions to obtain an angiopep-2 modified chitosan solution.

[0050] (3) The pH of the chitosan solution modified by angiopep-2 is adjusted to 5.0, 10 mL of the above solution is taken, 5 mg of oxytocin is added thereto, and 3.96 mL of sodium phosphate tr...

Embodiment 3

[0056] This example evaluates the efficacy of the oxytocin-loaded chitosan nanogel prepared in Example 2 for early intervention in Alzheimer's disease.

[0057] (1) Cell level test:

[0058] The microglial cell line BV-2 cells were used as the experimental object, and the Alzheimer's disease cell model with inflammatory response was constructed by lipopolysaccharide stimulation. 5 Cells were seeded in a 24-well plate at a density of 37°C for 24 hours. When the cells grew to about 80%, 1 mL of medium containing 50 μg / mL lipopolysaccharide was added and incubated for 24 hours.

[0059] The microglial cell line BV-2 cells were used as the experimental object, and the early cell model of Alzheimer's disease was constructed by pretreatment of oxytocin-loaded chitosan nanogel. 10 5 Cells were seeded in a 24-well plate at a density of 37°C for 24 hours. When the cells grew to about 80%, 1 mL of medium containing 100 μg / mL oxytocin-loaded chitosan nanogel was added to incubate for 2...

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Abstract

The invention relates to an application of chitosan nanogel loaded with oxytocin in preparation of a medicine for early intervention of Alzheimer's disease. The research finds that the chitosan nanogel loaded with oxytocin has a remarkable effect in the aspect of early treatment of the Alzheimer's disease. Cellular level experiment results show that the chitosan nanogel loaded with the oxytocin can inhibit microglial cell activation caused by lipopolysaccharide and reduce expression of inflammatory factors in microglial cells. Animal level experiment results show that the chitosan nanogel loaded with the oxytocin can significantly reduce the expression level of inflammatory factors of hippocampus of an early model mouse with the Alzheimer's disease, significantly reduce beta-amyloid protein deposition and neuron damage, and significantly improve the learning and memory ability of the early model mouse with the Alzheimer's disease. The chitosan nanogel loaded with the oxytocin is simple in preparation process, mild in reaction condition and easy in surface modification.

Description

technical field [0001] The invention belongs to the technical field of biomedicine, and relates to a new application of chitosan nanogel loaded with oxytocin, in particular to a kind of chitosan nanogel loaded with oxytocin to treat Alzheimer's disease through anti-inflammatory effect. Application of early intervention. Background technique [0002] Alzheimer's disease (AD) is a common neurodegenerative disease characterized by progressive memory loss and cognitive decline. β-Amyloid (Aβ) deposition and hyperphosphorylation of tau protein are considered to be the main pathological features of AD, but early clinical interventions for these two basically ended in failure. Microglia-mediated chronic inflammation occurs early in AD and precedes the accumulation of the above two pathogenic proteins. As the innate immune response is continuously triggered to restore tissue homeostasis, microglia are hyperactivated and continuously release a large number of inflammatory mediators...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K38/095A61K9/06A61K47/36A61K47/62A61P25/28A61P25/00
CPCA61K38/095A61K9/06A61K47/36A61K47/62A61P25/28A61P25/00
Inventor 窦妍叶才华汪俊萍于春水
Owner TIANJIN MEDICAL UNIV
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