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Preparation method of diacylglycerol kinase alpha gene-chitosan nanoparticles

A technology of chitosan nanoparticles and ester kinase, applied in gene therapy, pharmaceutical formulations, respiratory diseases, etc., can solve the problem of weakening T cell activation signal CD69 expression, T cell anergy, weakening Ras activation signal and inhibiting ERK phosphorylation And other issues

Inactive Publication Date: 2014-08-27
THE SECOND AFFILIATED HOSPITAL ARMY MEDICAL UNIV
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  • Abstract
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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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  • Preparation method of diacylglycerol kinase alpha gene-chitosan nanoparticles
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  • Preparation method of diacylglycerol kinase alpha gene-chitosan nanoparticles

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Embodiment

[0042] 1. Preparation of DGKα (diglyceride kinase α) gene-chitosan nanoparticles:

[0043] Step 1. The fusion gene of the mouse IL-2 signal peptide sequence and the mouse DGKα gene was synthesized completely, and cloned into the pEGFP-N3 vector to obtain the DGKα plasmid. Sequencing confirmed that the mouse IL-2 signal peptide and mouse DGKα gene sequences were completely consistent with GenBank's IL-2 (NM_008366.3) and DGKα gene (NM_016811.2);

[0044] Step 2, dissolving chitosan in a solution of 1% acetic acid concentration to prepare a chitosan acetic acid solution with a concentration of 1.5 mg / ml, adjust its pH value to 5.5 with sodium hydroxide, and then use a pore size of 0.22 μm sterile membrane filtration sterilization, in which chitosan has a molecular weight of 100,000 Daltons and a deacetylation degree of 95%;

[0045] Step 3, dissolving the DGKα plasmid in the anhydrous sodium sulfate solution filtered by a sterile membrane of 0.22 μm of 10 mM to prepare a DGKα s...

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Abstract

The invention discloses a preparation method of diacylglycerol kinase alpha gene-chitosan nanoparticles. The preparation method is characterized in that specific to the disadvantages that naked DGK alpha plasmids are easy to destroy by nucleases, acids, bases and the like in bodies, and the DGK alpha gene-chitosan nanoparticles are prepared by using chitosan and are capable of effectively protecting the DGK alpha plasmids, enhancing the activity of the DGK alpha plasmids in the bodies and promoting a DGK alpha gene to work in the bodies relatively well. The DGK alpha has the effect of inducing T-cell anergy and can induce immune tolerance. The DGK alpha gene-chitosan nanoparticles prepared by the method can inhibit airway inflammation of asthma rats induced by egg albumin and reduce the specific IgE (immunoglobulin E) level of serum egg albumin and is expected to be applied to treatment of allergic diseases such as asthma.

Description

[0001] The present invention is a divisional application for the invention of "A Diacylglycerol Kinase α Gene-Chitosan Nanoparticles Used in the Preparation of Drugs for Treating Allergic Asthma". (the application date of the original application document: February 19, 2013, the application number of the original application document: 201310053559.6, the name of the original application document: a kind of diglyceride kinase α gene-chitosan nanoparticle in the preparation of treatment of allergic asthma drug application) technical field [0002] The invention belongs to the field of biomedical preparations, in particular to a method for preparing diglyceride kinase alpha gene-chitosan nanoparticles. Background technique [0003] Asthma is a chronic airway allergic inflammatory disease, CD4 + The overactivation of T cells plays an important role in the pathogenesis of asthma. CD4 + After T cell activation, the differentiation into Th2 cells increases, and the excessive act...

Claims

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Application Information

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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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