Application of diacylglycerol kinase alpha gene-chitosan nanoparticle in preparing medicine for treating allergic asthma

A chitosan nanoparticle, allergic asthma technology, applied in gene therapy, allergic diseases, drug combination, etc., can solve the problem of weakening T cell activation signal CD69 expression, T cell anergy, weakening Ras activation signal and inhibiting ERK phosphorylation issues such as

Inactive Publication Date: 2013-05-22
THE SECOND AFFILIATED HOSPITAL ARMY MEDICAL UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

Sanjuan et al [Sanjuan MA, Jones DR, Izquierdo M, et al.Role of diacylglycerol kinase alpha in the attenuation of receptor signaling.J Cell Biol.2001:153,207-220.Sanjuan MA, Pradet-Balade B, Jones DR, et al. T cell activation in vivo targets diacylglycerol kinase to the membrane: a novel mechanism for Ras attenuation. J Immunol, 2003, 170: 2877-2883.] The study found that T cells transfected with DGKα gene overexpressed DGKα and inhibited diacylglycerol Ester-dependent T cell receptor (TCR) signal, thereby weakening Ras activation signal and inhibiting ERK phosphorylation, inhibiting the activity of IL-2 promoter, weakening the expression of T cell activation signal CD69, resulting in T cell anergy
At present, there are no related reports on the preparation of DGKα gene-chitosan nanoparticles, and reports on the application of chitosan-wrapped DGKα gene in the treatment of asthma

Method used

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  • Application of diacylglycerol kinase alpha gene-chitosan nanoparticle in preparing medicine for treating allergic asthma
  • Application of diacylglycerol kinase alpha gene-chitosan nanoparticle in preparing medicine for treating allergic asthma
  • Application of diacylglycerol kinase alpha gene-chitosan nanoparticle in preparing medicine for treating allergic asthma

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Embodiment

[0041] 1. Preparation of DGKα (Diglyceride Kinase α) Gene-Chitosan Nanoparticles:

[0042] Step 1. Fully synthesize the fusion gene of mouse IL-2 signal peptide sequence and mouse DGKα gene and clone into pEGFP-N3 vector to obtain DGKα plasmid. Sequencing confirmed that the mouse IL-2 signal peptide and the mouse DGKα gene sequence are completely consistent with the IL-2 (NM_008366.3) and DGKα gene (NM_016811.2) of GenBank;

[0043] Step 2. Dissolve chitosan in a solution with an acetic acid concentration of 1% to prepare a chitosan acetic acid solution with a concentration of 1.5 mg / ml, adjust its pH to 5.5 with sodium hydroxide, and then use a pore size of 0.22 μm sterile membrane filtration sterilization, in which the molecular weight of chitosan is 100,000 Daltons, and the degree of deacetylation is 95%;

[0044] Step 3. Dissolve the DGKα plasmid in 10mM 0.22μm sterile membrane-filtered anhydrous sodium sulfate solution to prepare a DGKα solution with a concentration of 360μg / ml...

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Abstract

The invention discloses an application of a DGK alpha (diacylglycerol kinase alpha) gene-chitosan nanoparticle as a unique active component in preparing a medicine for treating allergic asthma as well as a preparation method of the DGK alpha gene-chitosan nanoparticle. The invention, aiming at shortcomings that naked DGK alpha plasmid is easy to be damaged by nuclease, acid, alkali and the like in the body, prepares the DGK alpha gene-chitosan nanoparticle through chitosan, so as to effectively protect the DGK alpha plasmid, enhance the activity of the DGK alpha plasmid in the body and promote the DGK alpha gene to develop better function in the body. The DGK alpha, based on a function of induction of T cell anergy, can induce immune tolerance. The DGK alpha gene-chitosan nanoparticle prepared by the invention can inhibit egg albumin-induced airway inflammation of asthmatic mice, reduce serum egg albumin specific IgE level, and is expected to be used for treating asthma and other allergic diseases.

Description

Technical field [0001] The invention belongs to the field of biomedical preparations, and particularly relates to the application of diglyceride kinase alpha (DGKα) gene-chitosan nanoparticles in the preparation of medicines for treating allergic asthma. Background technique [0002] Asthma is a chronic airway allergic inflammatory disease, and the excessive activation of CD4+T cells plays an important role in the pathogenesis of asthma. After activation of CD4+ T cells, the differentiation into Th2 cells increased, and the overactivation of Th2 cells resulted in Th2 response-based immune response, and increased secretion of Th2 cytokines IL-4, IL-5 and IL-13. , Eventually leading to the accumulation of inflammatory cells such as eosinophils in the airway, causing airway allergic inflammation. Studies have shown that after normal people are repeatedly exposed to allergens, the stimulation of the allergens will not cause excessive activation of antigen-specific T cells to initiat...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K48/00A61K47/36A61P11/06A61P37/08
Inventor 王彦王长征林科雄程晓明
Owner THE SECOND AFFILIATED HOSPITAL ARMY MEDICAL UNIV
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