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Use of Triperugium wilfordii monomeric compound for increasing gene expression efficiency mediated by adeno-associated virus vector and subsidiarily treating neurodegenerative disease

A viral vector, Tripterygium wilfordii technology, applied in the field of Tripterygium wilfordii monomer compounds in the field of adjuvant treatment of neurodegenerative diseases, can solve problems such as low transduction efficiency, and achieve the effects of saving expensive costs, improving safety, and reducing the risk of immune response

Active Publication Date: 2008-10-22
CAPITAL UNIVERSITY OF MEDICAL SCIENCES
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0011] The technical problem to be solved by the present invention is to provide a method that can effectively improve the transduction efficiency of AAV in nerve cells in view of the relatively low transduction efficiency of AAV as a gene therapy carrier for central nervous system degenerative diseases. drug

Method used

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  • Use of Triperugium wilfordii monomeric compound for increasing gene expression efficiency mediated by adeno-associated virus vector and subsidiarily treating neurodegenerative disease
  • Use of Triperugium wilfordii monomeric compound for increasing gene expression efficiency mediated by adeno-associated virus vector and subsidiarily treating neurodegenerative disease
  • Use of Triperugium wilfordii monomeric compound for increasing gene expression efficiency mediated by adeno-associated virus vector and subsidiarily treating neurodegenerative disease

Examples

Experimental program
Comparison scheme
Effect test

experiment example 1

[0057] Experimental Example 1 Different concentrations of T 10 Experiments on the effect of rAAV2 on transduction efficiency in SH-SY5Y cells

[0058] To explore T 10 The effect of rAAV2 transduction efficiency in neural cell lines, using different concentrations of T 10 Add the SH-SY5Y cells together with the virus solution for incubation, and detect the activity of the reporter gene-luciferase carried by the virus after 24 hours. Because the MOI is the same and the action time and conditions are the same, if the luciferase activity detected in the cell fluid is high, it proves that the concentration of T 10 It has an enhancing effect on the transduction efficiency of rAAV2.

[0059] 1. Experimental method

[0060] The dopaminergic cell line SH-SY5Y cells were selected as a representative, with 1×10 4 Density per well Cells were seeded in 96-well plates. with different T 10 The cells were divided into 5 groups according to the treatment concentration: 0nM, 12.5nM, 25nM...

experiment example 2

[0063] Experimental Example 2 T of different action time courses 10 Experiments on the effect of rAAV2 on transduction efficiency in SH-SY5Y cells

[0064] To optimize T 10 The time of action, the time points of pretreatment, whole process treatment and short-term action after infection were respectively added in T 10 effect, the detected luciferase activity was compared with that without T 10 Comparison of treatment groups, and comparison between different treatment time groups, to see which group has a better effect on enhancing luciferase activity. Experimental results confirm that: adding T 10 It has the effect of enhancing the transduction efficiency, and the effects of different action time courses are also different, and the 24-hour full-course action group has the best effect.

[0065] 1. Experimental method

[0066] The dopaminergic cell line SH-SY5Y cells were selected as a representative, with 1×10 4 Density per well Cells were seeded in 96-well plates. at di...

experiment example 3

[0069] Experimental Example 3 Different concentrations of T 10 Experiments on the effect of rAAV2 on transduction efficiency in MN9D cells

[0070] To confirm T 10 Whether the increase of rAAV2 transduction efficiency is universal, this experiment took the MN9D cell line as the target cell (dopaminergic cell) to conduct a verification experiment. The experimental results confirmed that T 10 All have enhanced effects on the luciferase activity of neuronal cell lines, and their effective concentration ranges are slightly different. T 10 Generalized enhancement of rAAV2 transduction in neuronal cells.

[0071] 1. Experimental method

[0072] The dopaminergic cell line MN9D cells were selected, and the MN9D cells were divided into 1×10 4 Cells were seeded in 96-well plates at a density per well. with different T 10 The cells were divided into 7 groups according to the treatment concentration: 0nM, 3.1nM, 6.2nM, 12.5nM, 25nM, 50nM and 100nM groups, and each group contained ...

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Abstract

The invention discloses the application of thunder god vine monomeric compound in improving the efficiency of the adeno-associated virus vector mediated gene expression, and also discloses the application of the thunder god vine monomeric compound in the adjuvant therapy for neurodegenerative diseases. The thunder god vine monomeric compound, in particular to Triptolide, can remarkably improve the transduction efficiency of 2-typed recombinant adeno-associated virus on a plurality of nerve cell systems, primary nerve cells and midbrain nervous tissues of rats. In addition, the thunder god vine monomeric compound can inhibit the activation of microglia, reduce the production of proinflammatory cytokine, increase the generation of neurotrophic factors and have protection effect on dopaminergic neuron and can be used for adjuvant therapy for neurodegenerative diseases. The invention effectively overcomes the bottleneck problem in the gene treatment of AAV-mediated neuropathy, fully combines the protection effect of the thunder god vine monomeric compound on nerve cells with the assistant synergy effect on virus vector and reduces the potential immune side effect and application cost.

Description

technical field [0001] The present invention relates to a new medical application of a monomeric compound of traditional Chinese medicine, in particular to the use of a monomeric compound of Tripterygium wilfordii Hook.f. in improving the gene expression efficiency mediated by an adeno-associated virus vector. The present invention also relates to The use of tripterygium wilfordii monomer compound in the auxiliary treatment of neurodegenerative diseases belongs to the field of biomedicine. Background technique [0002] Parkinson's disease (PD) is a degenerative disease of the central nervous system that commonly occurs in middle-aged and elderly people. In my country, its incidence rate is increasing year by year. The main pathological feature of PD is the progressive degeneration of dopaminergic neurons in the substantia nigra compacta, resulting in decreased dopamine content in the striatum. Its typical clinical manifestations are tremor, muscle stiffness and bradykinesi...

Claims

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Application Information

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IPC IPC(8): A61K31/585A61K48/00A61P25/28
Inventor 王晓民胡静丁卫张婷
Owner CAPITAL UNIVERSITY OF MEDICAL SCIENCES
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