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A kind of pharmaceutical composition and its application

A composition and drug technology, applied in the direction of drug combination, drug delivery, pharmaceutical formulation, etc., can solve the problems of lack of space and tissue specificity, neuron degeneration, death, etc., and achieve the effect of long duration of drug action and low toxicity

Active Publication Date: 2020-02-07
SHENZHEN INST OF ADVANCED TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

This type of method lacks spatial and tissue specificity. 6-hydroxydopamine is a selective TH neuron chemical damage agent. When injected into animals, the membrane transporters of neuron terminals or cell bodies are actively taken up into the cells and oxidized to generate neurotoxicants. Such as hydroxyl free radicals and quinones, which can damage the antioxidant system of TH neurons, damage mitochondrial function, membrane stability and DNA integrity, thereby causing neuron degeneration and death, and causing irreversible damage to the peripheral sympathetic nervous system. damage

Method used

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  • A kind of pharmaceutical composition and its application
  • A kind of pharmaceutical composition and its application
  • A kind of pharmaceutical composition and its application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0060] (1) constructing an adeno-associated virus carrying a chemical genetic gene;

[0061] (1') Plasmid construction: construct the chemical genetic gene, red fluorescent gene and nerve cell-specific promoter on the pAAV plasmid to obtain pAAV-syn DIO-hM4Di-mCherry, the schematic diagram is as follows figure 1 shown;

[0062] (2') Transfection of cells: the plasmid obtained in step (1') and the viral packaging plasmid pHelper and pAAV-RC9 of the adeno-associated virus were co-transfected into 293FT cells, and the concentration of the transfection reagent was 5-8 mg / L per 200,000 cells of liposomes;

[0063] (3') Harvesting virus: change the medium 24 hours after transfection, the culture medium after the medium change is DMEM containing 5mM sodium pyruvate, perform ultracentrifugation to rupture the membrane 16 hours after the fluid change, the centrifugal speed is 5000g, and suspend the membrane after the membrane rupture The liquid was passed through a 20% sucrose filter...

Embodiment 2

[0068] (1) constructing an adeno-associated virus carrying a chemical genetic gene;

[0069] (1') Plasmid construction: the chemical genetic gene, red fluorescent gene and nerve cell-specific promoter were constructed on the pAAV plasmid to obtain pAAV-syn DIO-hM4Di-mCherry;

[0070] (2') Transfection of cells: the plasmid obtained in step (1') and the viral packaging plasmid pHelper and pAAV-RC9 of the adeno-associated virus were co-transfected into 293FT cells, and the concentration of the transfection reagent was 5 mg / L of lipid per 200,000 cells. plastid;

[0071] (3') Harvesting virus: change the medium 20 hours after transfection, the culture medium after the change is DMEM containing 4mM sodium pyruvate, and perform ultracentrifugation to rupture the membrane 12 hours after the fluid change, the centrifugal speed is 4000g, and suspend the membrane after the membrane is ruptured. The liquid was passed through a 20% sucrose filter column, and the supernatant was collecte...

Embodiment 3

[0076] (1) constructing an adeno-associated virus carrying a chemical genetic gene;

[0077] (1') Plasmid construction: the chemical genetic gene, red fluorescent gene and nerve cell-specific promoter were constructed on the pAAV plasmid to obtain pAAV-syn DIO-hM4Di-mCherry;

[0078](2') Transfection of cells: the plasmid obtained in step (1') and the viral packaging plasmid pHelper and pAAV-RC9 of the adeno-associated virus were co-transfected into 293FT cells, and the concentration of the transfection reagent was 8 mg / L of lipid per 200,000 cells. plastid;

[0079] (3') Harvesting virus: change the medium 30 hours after transfection, the culture medium after the medium change is DMEM containing 8mM sodium pyruvate, perform ultracentrifugation to rupture the membrane 20 hours after the fluid change, the centrifugal speed is 6000g, and suspend the membrane after the membrane rupture The solution was passed through a 20% sucrose filter column, and the supernatant was collected...

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Abstract

The invention discloses a pharmaceutical composition and an application thereof. The pharmaceutical composition comprises adeno-associated viruses carrying chemical genetic genes and / or medicines foractivating the chemical genetic genes. According to the pharmaceutical composition provided by the invention, an adeno-associated virus carrying chemical genetic genes is constructed to infect mice and target TH positive neuron cells in bone marrow cavities, noradrenaline secretion of the neuron cells can be specifically inhibited by drug activation, and the purposes of inhibiting sympathetic nerves and increasing bone mass safely and effectively for a long time are achieved. The pharmaceutical composition provided by the invention have the advantages of safety, stability, high efficiency andconvenience in application, has no toxicity to nerve cells, and has a remarkable action effect and a great application prospect and market value.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to a pharmaceutical composition and its application. Background technique [0002] The sympathetic nerve is an important type of nerve in the peripheral nervous system. The function of the sympathetic nerve plays an important role in the normal metabolism of the body and the control of glandular hormone secretion. Nerve fibers from the spinal cord go to the sympathetic ganglion, from which fibers are distributed to the viscera, cardiovascular system, and glands. Under normal circumstances, the sympathetic and parasympathetic nerves with opposite functions are in mutual balance and restriction. When the body is in a state of intense activity, sympathetic nerve activity plays a major role. The primary preganglionic neurons of the sympathetic nerve are located in the thorako-lumbales system of the spinal cord. These ganglia are interconnected into trunks called "sympathetic trunks." Po...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): A61K48/00A61K31/551A61K47/46A61P19/08A61P25/00
CPCA61K9/0019A61K31/551A61K47/46A61K48/0033A61K48/005
Inventor 高大双杨帆刘运辉
Owner SHENZHEN INST OF ADVANCED TECH