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Application of anti-EGFR scFv::FTH1/FTH1 protein nanoparticles in preparation of drugs

A nanoparticle and protein technology, applied in the field of nanobiomedicine, can solve the problems of unclear anti-EGFR antibody effect and unknown pathogenesis, and achieve the effects of reducing the number of goblet cells, expanding the cross-sectional area, and reducing airway resistance.

Inactive Publication Date: 2018-08-10
EAST CHINA UNIV OF SCI & TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Some studies in recent years have shown that epidermal growth factor receptor (EGFR) is involved in the proliferation of airway epidermal goblet cells and smooth muscle cells, which may be related to the occurrence and development of asthma. However, there are many factors involved in asthma, and its true The pathogenesis of asthma is unknown, so the role of anti-EGFR antibodies in the treatment of asthma has not been clarified

Method used

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  • Application of anti-EGFR scFv::FTH1/FTH1 protein nanoparticles in preparation of drugs
  • Application of anti-EGFR scFv::FTH1/FTH1 protein nanoparticles in preparation of drugs
  • Application of anti-EGFR scFv::FTH1/FTH1 protein nanoparticles in preparation of drugs

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0026] Example 1 Preparation and Characterization of Anti-EGFR scFv::FTH1 / FTH1 Protein Nanoparticles

[0027] Two plasmids, pET-28(+)-FTH1 and pET-28a(+)-anti-EGFR scFv-FTH1, exist in our laboratory (see ZL201010239499.3 for specific preparation methods). Wherein, the amino acid sequence of the anti-EGFR scFv::FTH1 protein is shown in SEQ ID NO.1, and the amino acid sequence of the FTH1 protein is shown in SEQ ID NO.2. Escherichia coli competent cells E.coli.BL21(DE3) and E.coli.DH5α were purchased from Beijing Tiangen Biochemical Technology Company. The above two plasmids were transformed into E.coli.DH5α competent cells, and positive clones were screened by colony PCR and enzyme digestion identification. Afterwards, the expression vector with the correct sequence was transformed into Ecoil.BL21 (DE3) competent cells by sequencing to obtain the engineered expression strain cells containing the FTH1 protein gene target engineering expression strain cells and the anti-EGFR scF...

Embodiment 2

[0033] Example 2 Effects of anti-EGFR scFv::FTH1 / FTH1 protein nanoparticles on the number of goblet cells in the lungs of mice and the amount of bronchial mucus secretion

[0034] 1. Experimental materials

[0035] 1.1 Experimental objects

[0036] SPF grade BALB / c mice, weighing 18-20 grams, aged 6-8 weeks, were randomly divided into 2 groups.

[0037] 1.2 Experimental reagents

[0038] 1) Intraperitoneal injection of sensitization solution containing ovalbumin (OVA) and Al(OH) 3 The PBS solution of OVA concentration is 10mg / L, and every mouse is intraperitoneally injected 0.2ml.

[0039] 2) The excitation solution is a PBS solution with an OVA concentration of 20mg / L, the excitation method is atomization excitation, the average median diameter of the atomized particles is 2.9 μm, and the atomization time is 30 minutes.

[0040] 3) Other reagents are obtained by preparing analytically pure drugs with sterile PBS.

[0041] 4) Anti-EGFR scFv::FTH1 / FTH1 protein nanoparticle...

Embodiment 3

[0052] Example 3 Effect of Anti-EGFR scFv::FTH1 / FTH1 Protein Nanoparticles on Collagen Fiber Deposition in Mouse Bronchial Epithelium

[0053] 1. Experimental method

[0054] 1) Asthma model construction was the same as in Example 2, that is, after the first, third, and seventh nebulizations, PBS and anti-EGFR scFv::FTH1 / FTH1 protein nanoparticles were injected intraperitoneally into the mice respectively. The mice were dissected 24 hours after the last injection, and the lung tissues of the mice were collected. Section staining was used to detect the deposition of collagen fibers under the bronchial epithelium, which is an important indicator of airway remodeling in bronchial asthma.

[0055] 2) The design of the experimental group, the technique of paraffin sectioning and the pre-treatment of section staining are the same as in Example 2.

[0056] 3) The treated tissue sections were stained with MASSON trichrome, the paraffin sections were dewaxed to water, and then washed...

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Abstract

The invention discloses application of anti-EGFR scFv::FTH1 / FTH1 protein nanoparticles in preparation of drugs, and in particular relates to application of the anti-EGFR scFv::FTH1 / FTH1 protein nanoparticles in preparation of drugs for preventing and treating asthma. The anti-EGFR scFv::FTH1 / FTH1 protein nanoparticles can significantly reduce the number of goblet cells in the lung of an asthmaticmodel mouse, reduce the amount of bronchial mucus secretion in the asthmatic model mouse and reduce the subcutaneous collagen fiber deposition in the asthmatic model mice, thus effectively expanding the cross-sectional area of the bronchus, reducing the airway resistance of the asthmatic mouse, and having preventing and treating effects on airway remodeling in the asthmatic mouse.

Description

technical field [0001] The invention belongs to the field of nano-biomedicine, and relates to the application of an anti-EGFR scFv::FTH1 / FTH1 protein nanoparticle in the preparation of medicines, in particular to an anti-EGFR scFv::FTH1 / FTH1 protein nanoparticle Application in the preparation of medicines for preventing and treating asthma. Background technique [0002] Asthma is a common disease. It is a chronic inflammatory disease of the airways involving multiple cells and cellular components, and this chronic inflammation is associated with airway hyperresponsiveness. The prominent pathological feature of asthma is the excessive secretion of airway mucus caused by airway goblet cell metaplasia or abnormal hyperplasia. At present, glucocorticoids are often used to control inflammation in the early stage of treatment, but recurrent asthma attacks usually cause airway remodeling, airway hyperplasia and narrowing, which makes the drug treatment effect worse in the later s...

Claims

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Application Information

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IPC IPC(8): A61K9/51A61K38/40A61K47/42A61P11/06
CPCA61K9/5169A61K38/40A61K47/42A61P11/06
Inventor 曹旭妮汪亮汪蓉蓉
Owner EAST CHINA UNIV OF SCI & TECH
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