CN108379240B

CN108379240B - Application of anti-EGFR scFv (monoclonal antibody against EGFR) FTH1/FTH1 protein nanoparticles in preparation of drugs

Info

Publication number: CN108379240B
Application number: CN201810196090.4A
Authority: CN
Inventors: 曹旭妮; 汪亮; 汪蓉蓉
Original assignee: East China University of Science and Technology
Current assignee: East China University of Science and Technology
Priority date: 2018-03-09
Filing date: 2018-03-09
Publication date: 2020-09-08
Anticipated expiration: 2038-03-09
Also published as: CN108379240A

Abstract

The invention discloses an application of anti (anti) EGFR scFv (human immunodeficiency Virus) FTH1/FTH1 protein nanoparticles in preparation of a medicament, in particular an application in preparation of a medicament for preventing and treating asthma. The anti-EGFRscFv comprises FTH1/FTH1 protein nanoparticles, wherein the FTH1/FTH1 protein nanoparticles can obviously reduce the number of lung goblet cells of an asthma model mouse, reduce the bronchial mucus secretion amount of the asthma model mouse and reduce the bronchial subcutaneous collagen fiber deposition of the asthma model mouse, so that the cross sectional area of a bronchus can be effectively enlarged, the airway resistance of the asthma model mouse is reduced, and the anti-EGFRscFv has a prevention and treatment effect on airway remodeling of the asthma model mouse.

Description

Application of anti-EGFR scFv (monoclonal antibody against EGFR) FTH1/FTH1 protein nanoparticles in preparation of drugs

Technical Field

The invention belongs to the field of nano biomedicine, and relates to an application of an anti (anti) EGFR scFv FTH1/FTH1 protein nano particle in preparation of a medicine, in particular to an application of an anti (anti) EGFR scFv FTH1/FTH1 protein nano particle in preparation of a medicine for preventing and treating asthma.

Background

Asthma is a common disease. It is a chronic inflammatory disease of the airways involving a variety of cells and cellular components, and this chronic inflammation is associated with airway hyperreactivity. A prominent pathological hallmark of asthma is excessive secretion of mucus at airway sites triggered by airway goblet cell metaplasia or abnormal hyperplasia. At present, glucocorticoid is often used for controlling inflammation in the early stage for treatment, but repeated asthma attacks usually cause airway remodeling and airway hyperplasia to cause stenosis, so that the treatment effect of the medicine is poor in the later stage. Among them, airway remodeling is one of the important indicators of asthma pathology, and its main pathological features are: goblet cell and mucus hyperplasia, bronchial subepithelial collagen fibrillation, airway smooth muscle hyperplasia hypertrophy, and epithelial alterations. The proliferation of goblet cells and the mucus secretion directly influence the cross-sectional area of an airway so as to reflect the size of airway resistance, and the deposition condition of collagen fibers under bronchial epithelium is an important index for reflecting airway remodeling of bronchial asthma, and the collagen fibers mainly consist of I type and III type. In recent years, studies 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 development of asthma, but many factors are involved in asthma, and the actual pathogenesis of the factors is unknown, so the role of anti-EGFR antibody in asthma treatment is not clear.

Single chain antibodies (scFv) are of particular interest as a genetically engineered antibody with a relatively small molecular weight. Because the molecular weight of the complete antibody is larger, the genetic engineering means is used for constructing the antibody fragment scFv with the antigen binding capacity. scFv contain only the portion of the antibody that binds to the antigen, and the overall size is only one-sixth of the full antibody, but still has some ability to bind to the target protein. The anti-epidermal growth factor receptor single-chain antibody (anti EGFR scFv) is a single-chain antibody capable of being specifically combined with EGFR, and the anti EGFR scFv can be used as a targeting molecule to mediate the specific combination of carrier protein and cells due to the expression of the EGFR on the surface of tissue cells. Based on the method, the anti-EGFR scFv is modified on the surface of the self-assembly protein of the ferritin heavy chain subunit (FTH1), so that the functionalized ferritin anti-EGFR scFv fusion protein with the target EGFR can be obtained, and the hybrid protein anti-EGFR scFv with the FTH1 protein can be further obtained, wherein the FTH1/FTH1 protein is obtained.

The protein FTH1/FTH1 belongs to nano particles, is a highly symmetrical biological macromolecule with a hollow cage-shaped structure and is self-assembled by 24 subunits, the protein nano particle is a protein constructed by the applicant at an early stage, and the specific sequence and the preparation method thereof are disclosed in a patent with the patent number ZL 201010239499.3. However, the patent does not mention the contents related to the treatment of asthma, so the invention further examines the effect of the protein on the treatment of asthma on the basis of the contents.

Disclosure of Invention

The invention aims to overcome the defect that the prior art lacks of medicaments for effectively controlling side effects such as airway remodeling, airway resistance enlargement and the like caused by asthma, and provides application of an anti-EGFR scFv (anti) FTH1/FTH1 protein nanoparticle in preparation of medicaments, in particular application in preparation of medicaments for preventing and treating asthma. The inventor finds that the anti-EGFR scFv protein nanoparticles can obviously reduce the number of lung goblet cells of an asthma model mouse, reduce the bronchial mucus secretion of the asthma model mouse and reduce the bronchial subcutaneous collagen fiber deposition of the asthma model mouse, thereby effectively enlarging the bronchial cross-sectional area of the asthma model mouse, reducing the airway resistance of the asthma model mouse and having prevention and treatment effects on airway remodeling of the asthma model mouse.

At present, anti-EGFR antibodies are mainly used for research related to tumor treatment, and because the functions in asthma prevention and treatment are uncertain as described above, and the mechanism of asthma is complex and unknown so far, the research related to asthma treatment is few. The applicant also carries out a plurality of researches on treating tumors on the basis of the fusion protein anti-EGFR scFv constructed in the early stage, namely FTH1 and the hybrid protein anti-EGFR scFv constructed in the early stage, namely FTH1/FTH1 protein nanoparticles; when the study on asthma is tried for the first time, the hybrid protein is surprisingly found to be capable of obviously reducing the number of lung goblet cells of an asthma model mouse, reducing the bronchial mucus secretion amount of the asthma model mouse and reducing the bronchial subcutaneous collagen fiber deposition of the asthma model mouse, so that the bronchial cross-sectional area of the asthma model mouse is effectively enlarged, the airway resistance of the asthma model mouse is reduced, and the hybrid protein has a prevention and treatment effect on airway remodeling of the asthma model mouse.

The invention solves the technical problem by applying the anti-EGFR scFv FTH1/FTH1 protein nanoparticles to the preparation of the medicine for preventing and treating asthma. The anti-EGFR scFv FTH1/FTH1 protein nanoparticle is a highly symmetrical biological macromolecule which is self-assembled into a hollow cage-shaped structure by 24 subunits, wherein the anti-EGFR scFv FTH1 protein subunit is formed by fusing the anti-EGFR scFv at the N end of FTH1 protein. The protein nanoparticle is a protein constructed in the early stage by the applicant, and the specific sequence and the preparation method thereof are disclosed in the patent with the patent number ZL 201010239499.3.

Preferably, the anti-EGFR scFv FTH1/FTH1 protein is formed by hybrid of anti-EGFR scFv FTH1 protein and FTH1 protein; wherein the amino acid sequence of the FTH1 protein is shown as SEQ ID NO.1, and/or the amino acid sequence of the FTH1 protein is shown as SEQ ID NO. 2; preferably, the anti-EGFR scFv is an FTH1 protein and/or an FTH1 protein which is a recombinant protein.

Preferably, the average particle size of the anti-EGFR scFv is 18nm +/-5 nm, wherein the FTH1/FTH1 protein nanoparticles have the average particle size.

Preferably, the molar ratio of the anti-EGFR scFv: FTH1/FTH1 protein nanoparticle to the anti-EGFR scFv: FTH1 protein to the FTH1 protein is less than or equal to 2, so that more target hybrid proteins which are folded correctly can be obtained; more preferably, the cost is balanced, and the molar ratio is 4: 6.

Preferably, the medicament for preventing and treating asthma is a medicament for reducing airway resistance and/or improving airway remodeling; more preferably, the medicament for preventing and treating asthma is a medicament for reducing the number of goblet cells, reducing the secretion amount of bronchial mucus and/or reducing the deposition of collagen fibers under bronchial epithelium. The goblet cell number and the bronchial mucus secretion directly influence the cross-sectional area of the airway so as to reflect the size of airway resistance, and the deposition condition of the collagenous fiber under bronchial epithelium is an important index for reflecting airway remodeling of bronchial asthma.

Preferably, the medicament comprises anti-EGFR scFv, FTH1/FTH1 protein nanoparticles and at least one medicinal carrier; more preferably, the anti-EGFR scFv comprises FTH1/FTH1 protein nanoparticles as the only active ingredient of the drug.

The drug comprises 0.1-99.9% of anti-EGFR scFv (Single chain variable fragment receptor) including FTH1/FTH1 protein nanoparticles and 0.1-99.9% of medicinal carriers, wherein the percentages are mass percentages.

The medicament of the invention is suitable for oral administration, intravenous, intramuscular, intradermal or subcutaneous administration and the like.

The medicine of the invention can be tablets, capsules, granules, injections or infusions and the like.

The medicament of the invention can be aqueous solution, non-aqueous solution or suspension, etc.

On the basis of the common knowledge in the field, the above preferred conditions can be combined randomly to obtain the preferred embodiments of the invention.

The reagents and starting materials used in the present invention are commercially available.

The positive progress effects of the invention are as follows: the invention applies the anti-EGFR scFv FTH1/FTH1 protein nanoparticles to the prevention and treatment of asthma, can obviously reduce the number of goblet cells, reduce the mucus secretion of bronchus and/or reduce the collagen fiber deposition under the epithelium of the bronchus, thereby effectively enlarging the cross-sectional area of the bronchus, reducing the airway resistance of an asthmatic mouse and having the prevention and treatment effect on the airway remodeling of the asthmatic mouse.

Drawings

FIG. 1 is NATIVE-PAGE schematic diagram of anti-EGFR scFv in which FTH1/FTH1 protein nanoparticles are renatured, wherein lane 1 is apoferritin, and lane 2 is anti-EGFR scFv in which FTH1/FTH1 protein nanoparticles are renatured.

FIG. 2 is a TEM schematic diagram of anti-EGFR scFv FTH1/FTH1 protein nanoparticles.

FIG. 3 is a fluorescent microscope representation of anti-EGFR scFv showing that FTH1/FTH1 protein nanoparticles bind to MDA-MB-468 cells, demonstrating that the nanoparticles can effectively bind to EGFR receptors.

FIG. 4 is a graph showing goblet cell proliferation and mucus secretion around bronchi of mice after PAS staining.

FIG. 5 is a schematic representation of collagen fiber deposition under mouse bronchial epithelium after MASSON trichrome staining.

Detailed Description

The invention is further illustrated by the following examples, which are not intended to limit the scope of the invention. The experimental methods without specifying specific conditions in the following examples were selected according to the conventional methods and conditions, or according to the commercial instructions.

Example 1 preparation and characterization of anti-EGFR scFv FTH1/FTH1 protein nanoparticles

Two plasmids, pET-28(+) -FTH1, pET-28a (+) -anti-EGFR scFv-FTH1, were present in the laboratory (see ZL201010239499.3 for details). The amino acid sequence of the FTH1 protein is shown as SEQ ID NO.1, and the amino acid sequence of the FTH1 protein is shown as SEQ ID NO. 2. Coli competent cells e.coli.bl21(DE3) and e.coli.dh5 α were purchased from beijing tiangen biochem technologies. The two plasmids are transformed into E.coli DH5 alpha competent cells, and positive clones are screened by colony PCR and enzyme digestion identification. Then, the expression vector with correct sequence is transformed into Ecoil. BL21(DE3) competent cells by sequencing to obtain target engineered expression strain cells containing FTH1 protein gene and anti-EGFR scFv:: FTH1 protein gene target engineered expression strain cells.

Next, the anti-EGFR scFv FTH1 protein gene target engineering expression strain cell expresses the anti-EGFR scFv FTH1 protein. The strain cells were cultured in LB medium containing kanamycin at 37 ℃. When the OD value of the culture solution was 0.4 to 0.6 (kanamycin concentration was 50. mu.g/ml), 1mM IPTG was added thereto to induce expression for 3 hours at 37 ℃. After induction, the cell pellet was centrifuged at 5000 rpm (20 minutes), washed once with Buffer A (50mM Tris-HCl, pH 7.9), and then disrupted by sonication. The cell lysate obtained by disruption was centrifuged at 5000 rpm (4 ℃ C., 15 minutes) to collect a precipitate, which was washed four times with Buffer B (50mM Tris-HCl, 50mM NaCl, 1mM EDTA, 1% Triton X-100, pH 7.9) to obtain inclusion bodies of anti-EGFR scFv:: FTH 1. FTH1 was then incubated in Buffer C containing 8M urea (50mM Tris-HCl, 8M urea, 1mM EDTA, 10mM DTT, pH 7.9) at 28 ℃ overnight to fully denature the protein.

FTH1 protein is expressed by FTH1 protein gene target engineering expression strain cells, and the expression conditions are the same as those for expressing anti-EGFR scFv: FTH1 protein. Except that FTH1 was obtained as a soluble protein. Here, purified FTH1 protein was obtained using Superose 6 size exclusion column AKTA (from GE). The FTH1 protein was then dissolved in Buffer C containing 8M urea and the protein was fully denatured overnight at 28 ℃. Then, the washed and denatured anti-EGFR scFv: FTH1 protein and FTH1 protein were mixed at a molar ratio of 4:6, the total mass of the proteins was 5mg, and the volume was 50 ml. Dialyzing sequentially at 4 ℃ against Buffer D containing 4, 3, 2, 1 and 0M urea, wherein Buffer D (50mM Tris-HCl, 0.5mM EDTA, 50mM NaCl, 10% glycerol, 0.1% polyethylene glycol (PEG), 0.5mM CuSO₄pH 7.9), then transferred into Buffer E (50mM Tris-HCl, 50mM NaCl, 10% glycerol, pH 7.9) for dialysis. Finally, the obtained protein is filtered and concentrated by a 50KD ultrafiltration device to obtain the high-concentration anti-EGFR scFv hybrid protein FTH1/FTH 1.

Non-denaturing polyacrylamide gel electrophoresis (as in FIG. 1) and Transmission Electron Microscopy (TEM) results (as in FIG. 2) show that the anti-EGFR scFv FTH1/FTH1 protein nanoparticles are correctly folded and assembled and the average diameter of the nanoparticles is 18nm +/-5 nm.

The fluorescent microscope detects the combination of the anti-EGFR scFv FTH1/FTH1 protein nanoparticles and MDA-MB-468 breast cancer cells (purchased from cell banks of Chinese academy of sciences) endogenously expressing EGFR on the cell surface, and as shown in FIG. 3, the anti-EGFR scFv FTH1/FTH1 protein has the activity of combining with the cell surface EGFR receptor.

The experimental result shows that the anti-EGFR scFv FTH1/FTH1 protein nanoparticle is successfully prepared and successfully characterized in a cell experiment, and the active anti-EGFR scFv FTH1/FTH1 protein nanoparticle is obtained.

Example 2 anti-EGFR scFv effects of FTH1/FTH1 protein nanoparticles on the number of goblet cells in the lungs and the amount of bronchial mucus secreted in the bronchi of mice

1. Experimental Material

1.1 test subjects

SPF-grade BALB/c mice, weighing 18-20 g, 6-8 weeks old, were randomly divided into 2 groups.

1.2 Experimental reagents

1) The sensitization liquid for intraperitoneal injection contains Ovalbumin (OVA) and Al (OH)₃The concentration of OVA in the PBS solution is 10mg/L, and 0.2ml is injected into the abdominal cavity of each mouse.

2) The excitation solution is PBS solution with OVA concentration of 20mg/L, the excitation mode is atomization excitation, the average median diameter is 2.9 μm, and the atomization time is 30 minutes.

3) All other reagents were prepared from analytically pure drug in sterile PBS.

4) anti-EGFR scFv FTH1/FTH1 protein nanoparticles prepared as described in example 1.

2. Experimental methods

Constructing an asthma model: injecting Ovalbumin (OVA) into abdominal cavity to sensitize BALB/c mouse, and performing OVA atomization excitation on BALB/c mouse 21 days later; the challenge was 1 time per day for 30 minutes for a total of 7 days. Wherein, PBS (control group) and anti-EGFR scFv nanoparticles are injected into the bodies of mice in the abdominal cavity respectively after the first, third and seventh atomization. Mice were dissected 24 hours after the end of the last injection and mouse lung tissue was collected. And (3) slicing and dyeing to detect goblet cell proliferation and mucus secretion around the bronchus of the lung, wherein the goblet cell proliferation and the mucus secretion directly influence the cross-sectional area of the airway and reflect the resistance of the airway.

The experimental groups were specifically divided into the following 2 groups:

A. PBS group: 3 weeks later, the OVA model mouse is injected with PBS (phosphate buffered saline) in an abdominal cavity with the volume of 0.2ml after the first, third and seventh atomization excitations.

B. FTH1/FTH1 proteome: the anti-EGFR scFv is intraperitoneally injected 3 weeks after the OVA model mouse, wherein the dose of the FTH1/FTH1 protein nano-particles is 0.75nM, the volume is 0.2ml, and the anti-EGFR scFv is injected after the first atomization excitation, the third atomization excitation and the seventh atomization excitation.

And after 24 hours of the last injection, the mice are killed by cervical dislocation, the mice are dissected, the whole lung is taken, 4% (w/v) paraformaldehyde is fixed for 24 hours, gradient alcohol dehydration is carried out after the fixation, the alcohol gradient is 50% (v/v), 70% (v/v), 85% (v/v), 95% (v/v) to 100% (v/v) pure alcohol in sequence, and each gradient dehydration lasts for 1 hour. After dehydration, the lung tissue was cleared, and then immersed in a mixture of pure alcohol and xylene at a volume ratio of 1:1 for 1.5 hours, and then immersed in pure xylene for 1 hour. And (3) after the tissue blocks are transparent, performing wax dipping and embedding, putting the tissue block in a mixed solution of molten paraffin and xylene in a volume ratio of 1:1 for 2 hours, sequentially transferring the tissue block into 2 molten paraffin solutions to dip for 3 hours respectively, performing wax dipping and embedding, and slicing the embedded tissue block after the embedded tissue block is immersed in a refrigerator at 4 ℃ overnight.

The treated tissue sections were stained with PAS, paraffin sections were dewaxed to water, and then placed in distilled water for 1 minute. 1 drop (about 100. mu.l) of periodic acid (reagent A) was added dropwise for 10 minutes, followed by rinsing with distilled water. 1 drop (about 100. mu.l) of Schiff's solution (reagent B) is added dropwise for dyeing for 10-20 minutes. Reagent B was decanted. 1 drop (100. mu.l) of sodium metabisulfite (reagent C) was added directly dropwise for 1 minute (2 times). Then rinsed under running water for 10 minutes. Gradient alcohol dehydration, xylene transparency, and neutral gum sealing. And (5) detecting the staining condition of the lung tissue cells of the mice by a microscope.

3. Results of the experiment

Periodic acid in PAS staining (periodic acid-Schiff staining) oxidizes intracellular polysaccharide glycol groups to dialdehydes, which combine with the colorless fuchsin of Schiff's solution, and the red color localizes to the cytoplasm. Since goblet cells and mucus secreted by goblet cells in lung tissue of the asthma model contain a large amount of glycogen, the goblet cells in lung tissue can be observed to be hyperplastic and hypertrophic and the secretion of mucus in bronchi through PAS staining. The proliferation of goblet cells and the amount of mucus secretion directly affect the cross-sectional area of the airway, and thus the effect on the magnitude of airway resistance is observed. As shown in FIG. 4, after PAS staining, mouse bronchial goblet cell proliferation was inhibited in the anti-EGFR scFv group compared with PBS group, and mucus secretion was significantly reduced in the FTH1/FTH1 protein nanoparticle group.

The experimental result shows that the anti-EGFR scFv protein nanoparticles FTH1/FTH1 have obvious inhibition on the goblet cell proliferation of an asthmatic mouse, and mucus secretion is obviously reduced, so that the bronchial cross-sectional area of the asthmatic mouse is effectively enlarged, and the airway resistance of the asthmatic mouse is effectively reduced.

Example 3 anti-EGFR scFv the Effect of FTH1/FTH1 protein nanoparticles on collagen fiber deposition under mouse bronchial epithelium

1. Experimental methods

1) The asthma model was constructed as in example 2, i.e., PBS and anti-EGFR scFv nanoparticles FTH1/FTH1 were injected intraperitoneally into mice after the first, third and seventh nebulizations, respectively. Mice were dissected 24 hours after the end of the last injection and mouse lung tissue was collected. The section staining detects the deposition of collagen fiber under bronchial epithelium, which is an important index for reflecting the airway remodeling of bronchial asthma.

2) Experimental group design, paraffin sectioning technique and pretreatment of section staining were the same as in example 2.

3) The treated tissue sections were subjected to MASSON trichrome staining, paraffin sections were dewaxed to water and then washed with tap water and distilled water in sequence for 5 minutes. Hematoxylin staining nuclei for 2 minutes, washing thoroughly, bluing with warm water, staining with MASSON trichrome ponceau acid re-reddish red for 10 minutes, differentiating with 1% (w/w) phosphomolybdic acid aqueous solution for 5 minutes, staining with 1% (w/v) aniline blue for 5 minutes, and finally differentiating with 1% (v/v) glacial acetic acid aqueous solution for 10 seconds. Gradient alcohol dehydration, xylene transparency, and neutral gum sealing. And (5) detecting the staining condition of the lung tissue cells of the mice by a microscope.

2. Analysis of Experimental results

MASSON trichrome staining is mainly used for staining and detecting collagen fibers. The lung tissue of the asthma model is subjected to airway remodeling due to a large amount of collagen deposition, and collagen fibers mainly consist of I type and III type and are bright red after MASSON trichrome staining. The experimental result is shown in figure 5, and the result shows that the deposition area of the anti-EGFR scFv mouse airway collagen fiber is obviously reduced compared with that of the PBS group by the FTH1/FTH1 protein nanoparticle group. The experimental result shows that the anti-EGFR scFv protein nanoparticles FTH1/FTH1 can obviously inhibit the deposition of airway collagen fibers of asthmatic mice, thereby having the prevention and treatment effect on airway remodeling of asthmatic mice.

While the invention has been described with respect to a preferred embodiment, it will be understood by those skilled in the art that the foregoing and other changes, omissions and deviations in the form and detail thereof may be made without departing from the scope of this invention. Those skilled in the art can make various changes, modifications and equivalent arrangements, which are equivalent to the embodiments of the present invention, without departing from the spirit and scope of the present invention, and which may be made by utilizing the techniques disclosed above; meanwhile, any changes, modifications and variations of the above-described embodiments, which are equivalent to those of the technical spirit of the present invention, are within the scope of the technical solution of the present invention.

SEQUENCE LISTING

<110> university of east China's college of science

Application of FTH1/FTH1 protein nanoparticles in preparation of drugs

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Claims

1. The application of the anti-EGFR scFv FTH1/FTH1 protein nanoparticles in preparing the medicine for preventing and treating asthma is characterized in that the anti-EGFR scFv FTH1/FTH1 protein is formed by heterozygosis of anti-EGFR scFv FTH1 protein and FTH1 protein; wherein the amino acid sequence of the FTH1 protein is shown as SEQ ID NO.1, and the amino acid sequence of the FTH1 protein is shown as SEQ ID NO. 2; the molar ratio of the anti-EGFR scFv in the anti-EGFR scFv-FTH 1/FTH1 protein nanoparticle to the FTH1 protein to the FTH1 protein is less than or equal to 2.

2. The application of the anti-EGFR scFv FTH1/FTH1 protein nanoparticle in preparing the medicine for preventing and treating asthma according to claim 1, wherein the anti-EGFR scFv FTH1 protein and/or FTH1 protein is recombinant protein.

3. The use of the anti-EGFR scFv FTH1/FTH1 protein nanoparticles in the preparation of a medicament for the prevention and treatment of asthma according to claim 1 or 2, wherein the average particle size of the anti-EGFR scFv FTH1/FTH1 protein nanoparticles is 18nm +/-5 nm.

4. The application of the anti-EGFR scFv: FTH1/FTH1 protein nanoparticle in the preparation of the drugs for preventing and treating asthma according to claim 1, wherein the molar ratio of the anti-EGFR scFv: FTH1/FTH1 protein nanoparticle to the anti-EGFR scFv: FTH1 protein to the FTH1 protein is 4: 6.

5. The anti-EGFR scFv according to claim 1, wherein the FTH1/FTH1 protein nanoparticle is used for preparing the medicine for preventing and treating asthma, and the medicine for preventing and treating asthma is used for reducing airway resistance and/or improving airway remodeling.

6. Use of anti-EGFR scFv nanoparticles FTH1/FTH1 protein for the manufacture of a medicament for the prevention or treatment of asthma according to claim 1, wherein the medicament for the prevention or treatment of asthma is a medicament for reducing the number of goblet cells, reducing the amount of bronchial mucus secretion and/or reducing the deposition of collagen fibers under the bronchial epithelium.

7. The use of the anti-EGFR scFv FTH1/FTH1 protein nanoparticles for the preparation of a medicament for the prevention and treatment of asthma as claimed in claim 1, wherein the medicament comprises the anti-EGFR scFv FTH1/FTH1 protein nanoparticles and at least one pharmaceutically acceptable carrier.

8. The application of the anti-EGFR scFv FTH1/FTH1 protein nanoparticles in preparing the medicine for preventing and treating asthma according to claim 7, wherein the anti-EGFR scFv FTH1/FTH1 protein nanoparticles are the only active ingredients of the medicine.