Peptide compositions and their use in the treatment or prevention of bronchial allergy-related diseases

Abstract

This invention discloses a peptide composition and its use in treating or preventing bronchial allergy-related diseases. The peptide composition disclosed herein comprises an effective amount of a polypeptide represented by any of the following amino acid sequences: SEQ ID No. 1 to SEQ ID No. 10. Since this polypeptide has the activity of reducing the imbalance of the ratio of interleukin-4, interleukin-13, and Th2, administering the peptide composition disclosed herein to an individual can effectively improve or prevent bronchial allergic reactions or related diseases.

Description

[Technical Field]

[0001] This invention relates to a functional peptide and its use, particularly to a peptide composition and its use in treating or preventing bronchial allergy-related diseases. [Previous Technology]

[0002] Note: Bronchial allergy, also known as tracheal sensitivity, refers to an allergic reaction caused by allergens irritating the trachea. This can lead to tracheal inflammation, swelling, phlegm buildup, or narrowing, and in severe cases, even difficulty breathing. Common allergens include dust mites, animal dander, airborne dust, pollen, etc. Asthma is a chronic bronchial inflammation causing respiratory allergies. Clinical symptoms include recurrent coughing, wheezing, shortness of breath, chest tightness, and difficulty breathing.

[0003] Statistics show that approximately 300 million people worldwide suffer from asthma, and about 180,000 people die from it each year. Asthma is a difficult disease to cure, and clinical treatment primarily involves medication. Asthma medications are mainly divided into symptom-controlling drugs and drugs used in the acute phase. Symptom-controlling drugs include inhaled and oral steroids, while drugs used in the acute phase include bronchodilators. Current research indicates that long-term use of oral steroids can cause side effects such as osteoporosis and moon face; while bronchodilators can cause side effects such as palpitations and tremors. Furthermore, using bronchodilators as a daily control medication not only fails to control asthma but also increases the risk of death for patients. [Summary of the Invention]

[0004] The main objective of this invention is to provide a peptide composition and its use in treating or preventing bronchial allergy-related diseases. It effectively improves bronchial allergy or the diseases it causes by administering a peptide that inhibits IL-4 and / or IL-13, thereby achieving the effects of daily health care and improving allergy symptoms.

[0005] Another object of the present invention is to provide a peptide composition and its use in treating or preventing bronchial allergy-related diseases. The peptide composition provided can effectively regulate an individual's immune response so as to serve as a means of daily health care, thereby reducing medication or lowering the frequency of drug use.

[0006] Therefore, in order to achieve the aforementioned objective, the present invention provides a peptide composition and its use in treating or preventing bronchial allergy-related diseases, wherein the peptide composition comprises at least one polypeptide, and the amino acid sequence of the polypeptide is any one of SEQ ID No. 1 to SEQ ID No. 10.

[0007] The peptide composition is a food, a nutritional supplement or a medicine.

[0008] Furthermore, the peptide composition disclosed in this invention can be used to inhibit the activity of IL-4 and / or IL-13 and reduce the concentration of IgE. Therefore, by administering an effective amount of the peptide composition disclosed in this invention to an individual, especially a person with bronchial allergy, it can effectively improve or alleviate bronchial allergy caused by allergens, such as asthma.

[0009] In one embodiment of the present invention, the peptide composition comprises at least the following four peptides: peptides shown in SEQ ID No. 4, SEQ ID No. 6, SEQ ID No. 8 and SEQ ID No. 9.

[0010] In another embodiment of the present invention, the peptide composition comprises the following peptides: SEQ ID No.1, SEQ ID No.2, SEQ ID No.3, SEQ ID No.4, SEQ ID No.5, SEQ ID No.6, SEQ ID No.7, SEQ ID No.8, SEQ ID No.9 and SEQ ID No.10.

[0011] In a further embodiment of the present invention, it is disclosed that any of the amino acid sequences SEQ ID No. 1 to SEQ ID No. 10 can be used to prepare a peptide composition that inhibits IL-4 and / or IL-13. Specifically, since the peptides SEQ ID No. 1 to SEQ ID No. 10 disclosed in the present invention have the activity of inhibiting IL-4 and / or IL-13, by administering an effective amount of any of the peptides SEQ ID No. 1 to SEQ ID No. 10 or a peptide composition containing such peptides to an individual, it is possible to effectively regulate the individual's immunity and reduce the chance of asthma occurrence.

Implementation Method

[0013] This invention discloses a peptide composition and its use in treating or preventing bronchial allergy-related diseases. Specifically, the peptide composition disclosed in this invention comprises an effective amount of a polypeptide represented by any of the following amino acid sequences: SEQ ID No. 1 to SEQ ID No. 10. Since this polypeptide has the activity of reducing the imbalance of the ratio of interleukin-4 (IL-4), interleukin-13 (IL-13), and Th2, administering the peptide composition disclosed in this invention to an individual can effectively improve or prevent an individual's allergic reactions or related diseases, especially bronchial allergy-related diseases such as asthma and allergic cough.

[0014] As explained herein, IL-4 and IL-13 are cytokines involved in the maintenance and exacerbation of chronic bronchial inflammation. Therefore, substances capable of inhibiting the activity of IL-4 and IL-13, such as the peptides disclosed in this invention or compositions containing such peptides, can effectively block downstream signaling pathways of IL-4 and IL-13, inhibiting inflammatory responses and airway hyperresponsiveness, thereby alleviating or improving the clinical symptoms of bronchial allergy. For example, dupilumab, a currently used asthma medication, is an IL-4Rα antagonist. This drug improves bronchial function and inflammatory markers in patients with moderate to severe asthma by inhibiting the IL-4 and IL-13 pathways.

[0015] Specifically, the polypeptides disclosed in SEQ ID No. 1 to SEQ ID No. 10 of the present invention are shown in Table 1 below. In Table 1, each capital letter in the amino acid sequence represents one amino acid. Unless otherwise stated, the amino acids represented by the capital letters are understood and interpreted based on common knowledge known to those skilled in the art.

[0016] Table 1: Peptide Sequences Serial Number amino acid sequence SEQ ID No.1 INFVTEEDKR SEQ ID No.2 LGVTNINISDP SEQ ID No. 3 RLTDYFTID SEQ ID No. 4 SMNLSLVLQVV SEQ ID No. 5 ALYPLLTTLTD SEQ ID No. 6 VRYLEWSNIESIIA SEQ ID No.7 LVGTDVATAQAP SEQ ID No. 8 GHCGCSSG SEQ ID No. 9 NFIVTGLQDIM SEQ ID No. 10 IGVDEHYTPI

[0017] In one embodiment of the present invention, the peptide composition comprises the peptide shown in SEQ ID No. 4, SEQ ID No. 6, SEQ ID No. 8 or SEQ ID No. 9.

[0018] In a further embodiment of the present invention, the peptide composition comprises the peptides shown in SEQ ID No. 4, SEQ ID No. 6, SEQ ID No. 8 and SEQ ID No. 9.

[0019] In another embodiment of the present invention, the peptide composition comprises the multiple peptides shown in SEQ ID No. 1 to SEQ ID No. 10.

[0020] In another embodiment of the present invention, the polypeptide is prepared by artificial synthesis or by a microbial production platform.

[0021] In another embodiment of the present invention, the polypeptide is obtained by separation, extraction, and / or purification from an animal milk, such as cow's milk, sheep's milk, or mare's milk. For example, the polypeptide is obtained by separation from sheep's milk through an extraction method, wherein the extraction method includes solvent extraction, supercritical fluid extraction, or other extraction methods known to those skilled in the art to which this invention pertains. Taking solvent extraction as an example, the extraction solvent used may be pure water, lower alcohol, acetone, ethyl acetate, or a combination of any two of the above solvents; and when the number of extraction solvents is large, extraction may be carried out sequentially with different extraction solvents according to the different solubilities of these extraction solvents.

[0022] The term "peptide" refers to a linear or cyclic molecule containing two or more amino acids, which can be isolated from substances of natural origin or prepared artificially, such as by chemical synthesis or recombinant microbial production platforms.

[0023] The term "composition" refers to a mixture containing two or more ingredients, including but not limited to active ingredients, excipients, solvents, carriers, buffers, and other pharmaceutically or food-acceptable ingredients. The composition may be in solid, liquid, gaseous, gel, emulsion, or any other dosage form and may be used for therapeutic, preventative, or daily health purposes. In the context of this invention, the composition may be a medicine, a food, or a nutritional supplement.

[0024] The term "effective dose" refers to a dose that produces the expected biological effect or therapeutic benefit, including but not limited to disease prevention, symptom relief, improvement of physiological parameters, or delay of disease progression. The effective dose may be adjusted according to factors such as the age, weight, condition, administration method and frequency of administration, and may be determined by clinical or pre-trial data.

[0025] The term "active ingredient" refers to a component in a composition that has the intended biological activity and can directly or indirectly induce the intended therapeutic, preventive, diagnostic, alleviating, improving, or controlling effects on a disease. Generally, the active ingredient accounts for 0.001-100% of the total weight of the composition. The active ingredient can be a naturally derived, semi-synthetic, or fully synthetic substance. In this invention, for example, the active ingredient is at least one peptide.

[0026] The term “introduction” means the introduction of an active ingredient or a composition containing the active ingredient into a living organism via any acceptable route, wherein the acceptable route includes, but is not limited to, oral, injection (e.g., intravenous, subcutaneous, intramuscular), inhalation, percutaneous, rectal or other medically acceptable routes, with the aim of producing a desired biological effect or therapeutic benefit in the living organism.

[0027] The term "bronchial hypersensitivity" refers to a physiological state in which the bronchi overreact to irritants such as cold air, allergens or chemicals, usually manifested as reversible airway constriction, airflow limitation or a tendency to have asthma attacks.

[0028] The term "asthma" refers to a chronic inflammatory respiratory disease characterized by at least one of the following symptoms: reversible airway obstruction, bronchial hypersensitivity, dyspnea, wheezing, or cough.

[0029] The term “one” or “at least one” means one or more quantities, and unless the context otherwise expressly limits, the term shall include the case of a single or multiple units.

[0030] The term "IL-4 (Interleukin-4)" is a key cytokine released by Th2 cells. It plays a central role in asthma and bronchial allergic inflammatory response and participates in the production of IgE immunoglobulin.

[0031] The term "IL-13 (Interleukin-13)" is a key cytokine released by Th2 cells. It plays a central role in asthma and bronchial allergic inflammatory response. It can promote airway epithelial remodeling, mucus secretion and bronchial smooth muscle hyperresponsiveness.

[0032] Hereinafter, in order to illustrate the technical features and effects of the present invention, several embodiments will be described in conjunction with the accompanying drawings.

[0033] In the following examples, the dosage of the disclosed composition in animal experiments may vary depending on factors such as the subject, dosage form, and form of the composition. The dosage variation is something that can be converted based on common knowledge in the technical field to which this invention pertains. For example, the dosage of the composition disclosed in this invention administered to mice is 45.05 mg / Kg, which is equivalent to a human dose of 3.65 mg / kg (approximately 219 mg / day) for a 60 kg adult.

[0034] Example 1: Preparation of multiple peptides

[0035] The multiple peptides shown in Table 1 above, SEQ ID No. 1-10, were prepared by artificial synthesis, and the amino acid sequences of each multiple peptide were confirmed to be correct.

[0036] Example 2: Cell Experiment

[0037] In this example, the cell line used was P815 mouse mastocytoma cell line, and the allergen was compound 48 / 80. The polypeptides disclosed in SEQ ID No. 1-10 of this invention were first co-cultured with P815 mouse mast cells for 20 minutes, and then compound 48 / 80 was added and co-cultured for 6 hours. After the culture was completed, the supernatant was collected, and the secretion levels of IL-4 and IL-13 were analyzed by ELISA. The results are shown in Figure 1 and Table 2.

[0038] As can be seen from the results in Figure 1 and Table 2, the polypeptide systems disclosed in SEQ ID No. 1-10 of the present invention have inhibitory activity against IL-4, and the polypeptide systems shown in SEQ ID No. 4, SEQ ID No. 5 and SEQ ID No. 9 have inhibitory activity against IL-13; and overall, the polypeptide systems shown in SEQ ID No. 4, SEQ ID No. 6, SEQ ID No. 8 and SEQ ID No. 9 have better activity in inhibiting the Th2 pathway.

[0039] It can be seen that the peptides disclosed in SEQ ID No. 1-10 of the present invention can respectively reduce the increase of IL-4 and / or IL-13 caused by allergic reactions and reduce the increase of allergic phenomena tending towards Th2 ratio, thereby improving the effects of inhaled allergic reactions, such as bronchial allergy, asthma, bronchitis and other diseases. In other words, by administering an effective amount of any of the peptides disclosed in SEQ ID No. 1-10 of the present invention to an individual suffering from bronchial allergy or related diseases, the symptoms of bronchial allergy can be effectively improved or alleviated. Among them, the improvement effect is better when administered to the peptides disclosed in SEQ ID No. 1, SEQ ID No. 6, SEQ ID No. 8 or SEQ ID No. 9.

[0040] Table 2: Inhibitory activity of various peptide sequences against IL-4 and IL-13 Serial Number IL-4 inhibitory activity (%) IL-13 inhibitory activity (%) SEQ ID No.1 -25.2 0.0 SEQ ID No.2 -16.0 0.0 SEQ ID No. 3 -8.9 0.0 SEQ ID No. 4 -32.9 -21.4 SEQ ID No. 5 -13.1 -71.4 SEQ ID No. 6 -22.2 -60.7 SEQ ID No.7 -23.9 0.0 SEQ ID No. 8 -38.5 0.0 SEQ ID No. 9 -11.5 -57.1 SEQ ID No. 10 -7.2 0.0

[0041] Example 3: Animal Experimentation

[0042] Mice were randomly divided into three groups. The experiment lasted for 23 days, and the treatment conditions for each group of mice were as follows:

[0043] Group 1 mice (blank group): These were normal mice that were not sensitized and were not given the peptide composition disclosed in this invention.

[0044] Group 2 mice (asthma group): These were asthma model mice, challenged with the virus from day 16 to day 22 of the experiment.

[0045] Group 3 mice (drug administration group): These were asthma model mice that were challenged with the drug from day 16 to day 22 of the experiment and were given the peptide composition disclosed in this invention daily during the experiment (dosage: 45.05 mg / Kg BW, where BW is body weight).

[0046] Among them:

[0047] Establishment of asthma pattern: On days 1, 8 and 15 of the experiment, OVA was administered to each mouse to sensitize them and induce respiratory inflammation symptoms.

[0048] Challenge treatment: From day 16 to day 22 of the experiment, mice were given PM2.5 daily (15 μg / mouse), and the PM2.5 samples were standard suspended particulate matter collected from the urban environment.

[0049] The active ingredient of the peptide composition disclosed in this invention comprises the multiple peptides shown in SEQ ID No. 1-10, and each of the multiple peptides is combined in equal or nearly equal proportions.

[0050] After the mice experiments in each group were completed, the airway resistance (Penh) values ​​of each group of mice were measured using a whole-body plethysmograph, and the area under the time-concentration curve (AUC) was calculated. The results are shown in Figures 2 and 3. Spleen cells were collected from each group of mice, and the proliferative activity of spleen cells and the activity of natural killer cells (NK cells) in each group of mice were detected. The results are shown in Figures 4 and 5. Bronchoalveolar lavage fluid (BALF) was collected from each group of mice, and the concentrations of IL-13, IL-4, and INF-γ in the BALF of each group of mice were detected by ELISA. The results are shown in Figures 6 to 8. In addition, the serum IgE and IgG1 concentrations of each group of mice were evaluated. The results are shown in Figures 9 and 10.

[0051] As can be seen from the results in Figures 2 and 3, the airway resistance of the mice in the third group was significantly lower than that of the mice in the second group, with a decrease of more than 30%, indicating that administration of the peptide composition disclosed in this invention can effectively improve asthma symptoms, that is, can alleviate asthma symptoms.

[0052] As shown in Figures 4 and 5, the splenocyte proliferation and natural killer cell activity of mice in Group 3 were increased compared with those in Group 2, indicating that administration of the peptide composition disclosed in this invention can prevent excessive activation of splenocytes and balance Th1 / Th2. In other words, the results in Figures 4 and 5 represent that the peptide composition of this invention helps improve allergic asthma, especially the Th2 over-activated type.

[0053] As shown in Figures 6 to 8, the concentrations of IL-4 and IL-13 in the BALF of the third group of mice decreased by 37.6% and 15.8% respectively compared with the second group of mice, and the concentration of INF-γ in the BALF of the third group of mice increased by 54.5% compared with the second group of mice; and as shown in Figures 9 and 10, the concentrations of IgE and IgG1 in the serum of the third group of mice decreased by 79.2% and 34.1% respectively compared with the second group of mice. The results of Figures 6 to 10 further demonstrate that the peptide composition disclosed in this invention can indeed regulate the expression of bronchial inflammatory cytokines, decrease the concentrations of IL-4 and IL-13 and increase the concentration of INF-γ, and significantly decrease the concentration of IgE in the serum, showing that the peptide composition disclosed in this invention does indeed have the effect of preventing or reducing the occurrence of allergic asthma and avoiding immune imbalance.

[0054] As can be seen from the above description, the peptides disclosed in SEQ ID No. 1-10 of the present invention have the activity of inhibiting IL-4 and / or IL-13 respectively. Therefore, administering an effective amount of the peptides disclosed in the present invention or a composition containing the peptides to an individual with bronchial allergy or at high risk can effectively prevent or improve diseases induced by bronchial allergy, such as asthma. [Simplified Explanation of the Diagram]

[0012] Figure 1 shows the results of detecting the inhibitory effect of various peptides on IL-4. Figure 2 shows the results of detecting the airway resistance (Penh) values ​​of mice in each group. Figure 3 shows the results of calculating the area under the curve of each group of mice in Figure 2. Figure 4 shows the results of analyzing the proliferation activity of spleen cells in each group of mice. Figure 5 shows the results of analyzing the activity of natural killer cells in each group of mice. Figure 6 shows the results of analyzing the concentration of IL-13 in bronchoalveolar lavage fluid (BALF) of mice in each group. Figure 7 shows the results of analyzing the concentration of IL-4 in bronchoalveolar lavage fluid of mice in each group. Figure 8 shows the results of analyzing the concentration of INF-γ in bronchoalveolar lavage fluid of mice in each group. Figure 9 shows the results of analyzing the concentration of IgE in the serum of mice in each group. Figure 10 shows the results of analyzing the concentration of IgG1 in the serum of mice in each group.

Claims

1. A polypeptide having the amino acid sequence shown in SEQ ID No.

6.

2. A peptide composition for inhibiting IL-4 and / or IL-13, comprising the multipeptide as described in claim 1.

3. Use of the polypeptide of claim 1 in preparing a peptide composition for treating or preventing bronchial allergic diseases, wherein, The peptide composition further comprises the peptides shown in SEQ ID No. 1-5 and SEQ ID No. 7-10.

4. The use as described in claim 3, wherein, This peptide composition is used to inhibit the activity of IL-4 and / or IL-13.

5. The use as described in claim 3, wherein, The peptide composition is a food, a nutritional supplement, or a pharmaceutical product.

6. The use as described in claim 3, wherein, This peptide composition is used to reduce the concentration of IgE.

7. The use as described in claim 3, wherein, This bronchial allergy is asthma.

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