Application of Anti-egfr scFv::FTH1/FTH1 Protein Nanoparticles in Drug Preparation

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 effect of reducing the number of goblet cells, expanding the cross-sectional area, and reducing airway resistance

Inactive Publication Date: 2020-09-08
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 Drug Preparation
  • Application of Anti-egfr scFv::FTH1/FTH1 Protein Nanoparticles in Drug Preparation
  • Application of Anti-egfr scFv::FTH1/FTH1 Protein Nanoparticles in Drug Preparation

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, 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 restriction enzyme identification. After that, the expression vector with the correct sequence was transformed into Ecoil.BL21(DE3) competent cells by sequencing to obtain the target engineering expression strain cells containing the FTH1 protein gene and the anti-EGFR scFv::FTH1 protein gene target engineering...

Embodiment 2

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

[0034] 1. Experimental materials

[0035] 1.1 Subject

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

[0037] 1.2 Experimental reagents

[0038] 1) Intraperitoneal injection of sensitizing solution, which contains ovalbumin (OVA) and Al(OH) 3 PBS solution, OVA concentration is 10mg / L, 0.2ml per mouse is injected intraperitoneally.

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

[0040] 3) All other reagents are prepared with sterile PBS for analytically pure drugs.

[0041] 4) Anti-EGFR scFv::FTH1 / FTH1 protein nanoparticles, prepared in Example 1.

[0042] 2. Experimental method

[0043] Ast...

Embodiment 3

[0052] Example 3 The effect of anti-EGFR scFv::FTH1 / FTH1 protein nanoparticles on the deposition of collagen fibers under the bronchial epithelium of mice

[0053] 1. Experimental method

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

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

[0056] 3) The processed tissue sections were stained with MASSON trichrome. After the paraffin sections were deparaffinized to water, they were washed with ...

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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 nanobiomedicine, and relates to the application of anti-EGFR scFv::FTH1 / FTH1 protein nanoparticles in the preparation of medicines, and specifically relates to an anti-EGFR scFv::FTH1 / FTH1 protein nanoparticles Application in preparing medicine for preventing and treating asthma. Background technique [0002] Asthma is a common disease. It is a chronic inflammatory disease of the airway involving a variety of cells and cell components. This chronic inflammation is related to airway hyperresponsiveness. The significant pathological manifestation of asthma is the excessive secretion of mucus in the airways caused by goblet cell metaplasia or abnormal proliferation of the airways. At present, glucocorticoids are often used to control inflammation in the early stage, but repeated attacks of asthma usually cause airway remodeling, airway hyperplasia and narrowing, which makes the drug treatment effect worse in the later st...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): A61K38/40A61K9/51A61K47/42A61P11/06
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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