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Application of dna tetrahedron in preparation of medicine for treating Alzheimer's disease

A technology for Alzheimer's disease and drug preparation, which is applied in the field of nerve repair and can solve the problem of less research on the effects of cell physiological activities.

Active Publication Date: 2021-04-23
CHENGDU TENGDASHU NANO BIOTECHNOLOGY CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Although there are many studies on TDNs, there are very few studies on its influence on various physiological activities of cells, especially the research on its treatment of Alzheimer's disease.

Method used

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  • Application of dna tetrahedron in preparation of medicine for treating Alzheimer's disease
  • Application of dna tetrahedron in preparation of medicine for treating Alzheimer's disease
  • Application of dna tetrahedron in preparation of medicine for treating Alzheimer's disease

Examples

Experimental program
Comparison scheme
Effect test

Embodiment

[0054] Synthesis of Example TDNs

[0055] 1. Synthesis

[0056] Four kinds of DNA single strands (S1, S2, S3, S4) are dissolved in the TM buffer solution with the same final concentration, wherein, the solute of the TMbuffer solution is Tris-HCl and MgCl , and their concentrations are respectively 10mM and 50mM, and adjusted The pH of the solution was 8.0. Then the mixture was vortexed, mixed, and centrifuged, then placed in a PCR instrument, and the temperature was rapidly raised to 95°C and stabilized for 10-15 minutes, then cooled to 4°C and stabilized for 20 minutes. That is synthesized as figure 1 The tetrahedral structure shown.

[0057] The sequence of the DNA single strand is shown in Table 1.

[0058] Table 1 Sequence of TDNs single strand

[0059]

[0060] 2. Identification

[0061] 2.1 Gel electrophoresis

[0062] A polyacrylamide gel was prepared by using 4.2mL distilled water, 1.2mL 40% acrylamide, 0.6mL 10×TAE, 60μL 10% Aps and 6μL TEMED;

[0063] Then...

experiment example 1A

[0072] Experimental example 1 Aβ external treatment of PC12 cells can simulate Alzheimer's disease

[0073] 1. Method

[0074] (1) Inoculate PC12 cell suspension (100 μL / well) in a 96-well plate, and place the culture plate in an incubator for pre-culture for 24 hours (37°C, 5% CO 2 ), then reduce the serum concentration in the culture medium consisting of DMEM+10% serum+1% double antibody from 10% to 6%, and cultivate it in an incubator for 6 hours (37°C, 5% CO 2 ), then reduce the serum concentration in the culture medium from 6% to 0, and cultivate in the incubator for 1 hour (37°C, 5% CO 2 ).

[0075] (2) Divide the cultured cell suspension into a control group and an experimental group, and add different concentrations of Aβ to the experimental group 25-35 , the control group was added with the same amount of serum, and then cultured in the incubator for 24h (37 ° C, 5% CO 2 ), after the end of the culture, add 10 μL of CCK-8 solution to each well, and avoid the produ...

experiment example 2

[0079] Experimental example 2 TDNs reduce the active damage of Aβ to PC12 cells

[0080] 1. Method

[0081] (1) Inoculate PC12 cell suspension (100 μL / well) in a 96-well plate, and place the culture plate in an incubator for pre-culture for 24 hours (37°C, 5% CO 2 ), then reduce the serum concentration in the culture medium consisting of DMEM+10% serum+1% double antibody from 10% to 6%, and cultivate it in an incubator for 6 hours (37°C, 5% CO 2 ), then reduce the serum concentration in the culture medium from 6% to 0, and cultivate in the incubator for 1 hour (37°C, 5% CO 2 ).

[0082] (2) Divide the cultured cell suspension into blank control group, TDNs group, AD cell model group and AD cell model group pretreated by TDNs; wherein, the blank control group does not add Aβ 25-35 ; The AD cell model group pretreated with TDNs was first pretreated with 250nM TDNs for 6h, and then added Aβ at a concentration of 25μM 25-35 ; AD cell model group was pretreated with serum-free ...

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Abstract

The present invention provides the use of DNA tetrahedron in the preparation of medicines for promoting nerve repair. The DNA tetrahedron is assembled into a tetrahedral structure by four single-stranded DNAs through complementary base pairing; each edge of the tetrahedron is double Strand DNA structure. The DNA tetrahedron of the invention can protect nerve cells in the environment of beta amyloid protein and inhibit their apoptosis. The invention has good application prospect in the preparation of Alzheimer's disease medicine.

Description

technical field [0001] The invention relates to the field of nerve repair, in particular to the use of DNA tetrahedrons in the preparation of drugs for treating Alzheimer's disease. Background technique [0002] Alzheimer's disease (AD) is one of the common neurodegenerative diseases, which is age-related. dementia. Studies have pointed out that apoptotic neuron death may be an important part of AD, neuron apoptosis has been found in the early stage of AD, and autopsies of AD patients also showed a large number of apoptotic neurons in the cerebral cortex and hippocampus . [0003] The pathogenesis of Alzheimer's disease is still unclear, and the amyloid-beta (Aβ) cascade hypothesis is widely influential. The hypothesis holds that the deposition of Aβ in the brain is the central link in the pathological changes of AD, which can trigger a series of pathological processes, which further promote the deposition of Aβ, thereby forming a cascade amplification reaction, and final...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): A61K31/7088A61P25/28
CPCA61K31/711
Inventor 林云锋邵晓茹蔡潇潇马文娟谢雪萍
Owner CHENGDU TENGDASHU NANO BIOTECHNOLOGY CO LTD
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