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Application of IL-1ra-Fcepsilon fusion gene and coded protein thereof to treatment on allergic rhinitis

A technology of 1.il-1ra-fc and 2.il-1ra-fc is applied in the application field of treating allergic rhinitis, and can solve the problems of adverse effects, short half-life and the like

Inactive Publication Date: 2012-10-10
INST OF BASIC MEDICAL SCI ACAD OF MILITARY MEDICAL SCI OF PLA
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, due to the extremely short half-life of IL-1ra in the body, and a large amount of IL-1ra is needed to eliminate the effect of IL-1, long-term high-dose application is necessary to achieve the therapeutic effect. Proven in arthritis, long-term high-dose systemic administration will have strong adverse effects

Method used

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  • Application of IL-1ra-Fcepsilon fusion gene and coded protein thereof to treatment on allergic rhinitis
  • Application of IL-1ra-Fcepsilon fusion gene and coded protein thereof to treatment on allergic rhinitis

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0021] Example 1 Preparation of IL-1ra-Fcε Fusion Gene Therapy Vector

[0022] The pIRES2-EGFP-IL-1ra-Fcε fusion gene expression vector [1] Transformed into Escherichia coli DH5α, cultivated in LB medium at 37°C vials, centrifuged at 9000rpm to collect bacteria, extracted a large amount of plasmids with the plasmid extraction kit from OMEGA company, concentrated the plasmids to more than 1mg / ml by ethanol precipitation, and stored at -20°C stand-by.

Embodiment 2

[0023] Example 2 Expression and protein purification of IL-1ra-Fcε fusion gene

[0024] The prokaryotic expression vector pBV220-IL-1ra-Fcε was transformed into Escherichia coli BL21, and the prokaryotic fermentation process was used to cultivate and induce Escherichia coli BL21 to express the recombinant protein. SPFF) to separate and purify the recombinant protein, and finally refold the protein by step-by-step dialysis [2] .

Embodiment 3

[0025] Embodiment 3 establishment of animal model [3，4]

[0026] Select healthy female Wistar rats weighing 135-165 g. On the first day, 1ml of normal saline, 1.5mg of egg protein, 3mg of aluminum hydroxide sol and 1×10 10 Inactivated pertussis bacilli were mixed and injected subcutaneously into the four paws of rats, and 0.2ml and 0.3ml were injected subcutaneously into each of the front and rear paws respectively. On the 5th day, 1ml of normal saline, 1.0mg of ovalbumin and 2mg of aluminum hydroxide sol were mixed and injected into the subcutaneous area of ​​the back, divided into 5 points, and each point was injected with 0.2ml for a boost. The mice were challenged from the 14th day: 1% ovalbumin physiological saline solution was instilled daily, 30 μl in each nostril, once a day, 10 times in total.

[0027] Modeling was successful if the behavior (sneezes and nose scratches) and symptoms (nasal secretions, etc.) within 30 minutes after nasal drip challenge were greater ...

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Abstract

The invention discloses application of IL-1ra-Fcepsilon fusion gene and coded protein thereof to treatment on allergic rhinitis, belonging to the field of medicines. In the invention, the constructed IL-1ra-Fcepsilon fusion gene expression vector is prepared into suspension according to a certain concentration and the suspension is directly dropped into a nasal cavity to treat the allergic rhinitis; or the purified IL-1ra-Fcepsilon fusion gene is diluted to a certain concentration with a PBS (Phosphate Buffered Saline) buffer solution and then the diluted solution is dropped into the nasal cavity to treat the allergic rhinitis. In the invention, the protein coded by the IL-1ra-Fcepsilon fusion gene can antagonize the inflammatory effect of IL-1 and inhibit mastocyte and basophilic granulocyte degranulation, thereby achieving the aim of treating the allergic rhinitis.

Description

technical field [0001] The invention belongs to the field of biomedicine and relates to the application of IL-1ra-Fcε fusion gene and its coded protein in treating allergic rhinitis. Background technique [0002] Nasal inflammatory cell infiltration and elevated serum IgE concentration are the main indications of allergic rhinitis. Inhibiting the increase of IgE level and anti-inflammatory treatment are research hotspots in the treatment of allergic rhinitis. Studies have confirmed that IL-1ra, as an effective anti-inflammatory factor, has the effect of treating and preventing allergic rhinitis, but the half-life of IL-1ra molecule is short, which limits its wide application to a certain extent. During the pathogenesis of rhinitis, the binding of IgE to its high-affinity receptor triggers the degranulation of mast cells and basophils, which is the key factor leading to the pathogenesis. Blocking the binding of IgE to its high-affinity receptor is one of the effective targe...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): A61K48/00A61K39/395A61P11/02A61P37/08
Inventor 徐东刚付文亮王园园邹民吉刘中成
Owner INST OF BASIC MEDICAL SCI ACAD OF MILITARY MEDICAL SCI OF PLA
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