Peptide composition and use thereof in treatment or prevention of bronchial hypersensitivity-related disease

Abstract

To provide a peptide composition and its use in treatment or prevention of bronchial hypersensitivity-related disease.SOLUTION: A peptide composition according to the present invention contains an effective amount of a polypeptide having a specific amino acid sequence. Since the polypeptide has an activity of reducing imbalance in interleukin-4, interleukin-13, and Th2 ratio, administration of the peptide composition to an individual provides an effect of effectively improving or preventing a tracheal allergic response or a related disease in the individual.SELECTED DRAWING: Figure 1

Description

[Technical Field]

[0001] The present invention relates to functional peptides and uses thereof, in particular to peptide compositions and their use in the treatment or prevention of bronchial hyperresponsiveness-related diseases. [Background technology]

[0002] Bronchial hyperresponsiveness, also known as reactive airway disease, is a phenomenon in which allergens irritate the trachea, triggering an allergic reaction. Symptoms include tracheal inflammation, swelling, phlegm accumulation, and narrowing, and in severe cases, can lead to difficulty breathing. Common allergens include dust mites, animal hair, airborne dust, particles, and pollen. Asthma is a respiratory disease caused by chronic inflammation of the bronchi, resulting in respiratory hypersensitivity. Clinical symptoms include recurrent coughing, wheezing, shortness of breath, chest tightness, and difficulty breathing.

[0003] According to statistics, approximately 300 million people worldwide suffer from asthma, and approximately 180,000 people die from it each year. Asthma is a disease that is difficult to cure, and the primary clinical treatment for asthma patients is drug therapy. Medications used to treat asthma are mainly divided into long-term control medications and acute relief medications. Long-term control medications include inhaled steroids and oral steroids, while acute relief medications include bronchodilators. Current research has shown that long-term use of oral steroids can cause side effects such as bone loss and lunacy, while bronchodilators can cause side effects such as palpitations and hand tremors. Using bronchodilators as a long-term control medication not only results in inability to control asthma, but may also increase the patient's risk of death. Summary of the Invention [Problem to be solved by the invention]

[0004] The main object of the present invention is to provide a peptide composition and its use in the treatment or prevention of diseases associated with bronchial hyperresponsiveness, which effectively improves bronchial hyperresponsiveness or diseases caused by it by administering a polypeptide that suppresses IL-4 and / or IL-13, thereby maintaining daily health and improving allergic symptoms.

[0005] Another object of the present invention is to provide a peptide composition and its use in the treatment or prevention of bronchial hyperresponsiveness-related diseases, which can effectively regulate an individual's immune response and, when used as a means of routine health management, can reduce the use of therapeutic drugs and the frequency of medication. [Means for solving the problem]

[0006] Therefore, to achieve the above-mentioned object, the present invention provides a peptide composition and its use in the treatment or prevention of bronchial hyperresponsiveness-related diseases, wherein the peptide composition comprises at least one polypeptide, the amino acid sequence of which is set forth in any one of SEQ ID NOs: 1 to 10.

[0007] The peptide composition can be a food, a dietary supplement, or a pharmaceutical.

[0008] Furthermore, the peptide composition of the present invention can be used to suppress 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 of the present invention to an individual, particularly a patient with bronchial allergy symptoms, it is possible to effectively improve or alleviate diseases associated with bronchial hyperresponsiveness caused by allergens, such as asthma.

[0009] In one embodiment of the invention, the peptide composition comprises at least the polypeptides 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 invention, the peptide composition comprises the polypeptides set forth in 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 a polypeptide consisting of an amino acid sequence represented by any one of SEQ ID NOs: 1 to 10 can be used to prepare a peptide composition that suppresses IL-4 and / or IL13. Specifically, the polypeptides represented by SEQ ID NOs: 1 to 10 disclosed in the present invention each have the activity of suppressing IL-4 and / or IL13, and therefore, by administering to an individual an effective amount of a polypeptide represented by any one of SEQ ID NOs: 1 to 10 or a peptide composition containing said polypeptide, it is possible to effectively regulate the individual's immunity and reduce the risk of developing asthma. [Brief explanation of the drawings]

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

[0013] The present invention discloses a peptide composition and its use in the treatment or prevention of bronchial hyperresponsiveness-related diseases. Specifically, the peptide composition of the present invention comprises an effective amount of a polypeptide consisting of an amino acid sequence represented by any one of SEQ ID NOs: 1 to 10. The polypeptide has the activity of reducing the imbalance of interleukin-4 (IL-4), interleukin-13 (IL-13), and the Th2 ratio. Therefore, administration of the peptide composition of the present invention to an individual can effectively improve or prevent the individual's allergic reactions or related diseases, particularly bronchial hyperresponsiveness-related diseases such as asthma and allergic cough.

[0014] More specifically, because IL-4 and IL-13 are cytokines involved in the maintenance and aggravation of chronic bronchitis, substances capable of inhibiting the activity of IL-4 and IL-13, such as the polypeptide of the present invention or a composition containing the polypeptide, can effectively inhibit downstream signaling of IL-4 and IL-13, suppressing inflammatory responses and airway hyperresponsiveness, thereby alleviating or ameliorating the clinical symptoms of bronchial allergy. For example, dupilumab, an existing asthma treatment, is an IL-4Rα antagonist that inhibits the IL-4 and IL-13 pathways, thereby improving bronchial function and inflammatory indicators in patients with moderate to severe asthma.

[0015] Specifically, the polypeptides disclosed in the present invention as shown in SEQ ID NO: 1 to SEQ ID NO: 10 are shown in Table 1 below, and each capital letter in the amino acid sequences shown in Table 1 represents one amino acid, and unless otherwise specified, the amino acids represented by each capital letter are understood and interpreted based on the common technical knowledge of those skilled in the art.

[0016] [Table 1]

[0017] In one embodiment of the invention, the peptide composition comprises a peptide set forth in SEQ ID NO:4, SEQ ID NO:6, SEQ ID NO:8 or SEQ ID NO:9.

[0018] In a next embodiment of the 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 polypeptides shown in SEQ ID NO:1 to SEQ ID NO:10.

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

[0021] In another embodiment of the present invention, the polypeptide is isolated, extracted, and / or purified from animal milk, such as cow's milk, sheep's milk, or horse's milk. For example, the polypeptide is isolated from sheep's milk by extraction. The extraction method may include solvent extraction, supercritical fluid extraction, or other extraction methods commonly known to those skilled in the art. For example, in the case of solvent extraction, the extraction solvent used may be pure water, a lower alcohol, acetone, ethyl acetate, or a combination of any two of the above solvents. When multiple extraction solvents are used, sequential extraction with different extraction solvents may be performed depending on the differences in solubility of the extraction solvents.

[0022] The term "polypeptide" refers to a linear or cyclic molecule containing two or more amino acids, which can be isolated from naturally occurring materials or prepared by artificial methods, such as 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 the form of a solid, liquid, gas, gel, emulsion, or any other dosage form, and may be used for treatment, prevention, or daily health maintenance. In the present invention, the composition may be a medicine, food, or nutritional supplement.

[0024] The term "effective amount" refers to a dose that produces a desired biological or therapeutic effect, including, but not limited to, disease prevention, symptom relief, improvement of physiological parameters, delay of disease progression, etc. The effective amount is adjusted depending on factors such as the subject's age, weight, and condition, administration method and frequency, and can be determined based on data from clinical trials and preliminary studies.

[0025] The term "active ingredient" refers to a component in a composition that has a desired biological activity and can directly or indirectly induce the desired treatment, prevention, diagnosis, mitigation, improvement, or control of a disease. Generally speaking, the active ingredient comprises 0.001 to 100% of the total weight of the composition. The active ingredient may be naturally occurring, semi-synthetic, or fully synthetic. For example, in the present invention, the active ingredient is at least one polypeptide.

[0026] The term "administration" means introducing an active ingredient or a composition containing said active ingredient into an organism via any acceptable route, including but not limited to oral, injection (intravenous, subcutaneous, intramuscular, etc.), inhalation, transdermal, rectal, or other medically acceptable route, for the purpose of producing a desired biological or therapeutic effect in said organism.

[0027] The term "bronchial hyperresponsiveness" refers to a physiological condition in which the bronchi are overly reactive to irritants such as cold air, allergens, or chemicals, and is usually manifested as reversible airway narrowing, airflow limitation, or a tendency to asthma attacks.

[0028] The term "asthma" refers to a chronic inflammatory airway disease characterized by at least one symptom including reversible airway obstruction, bronchial hyperresponsiveness, dyspnea, wheezing, coughing, and the like.

[0029] The terms "one" or "at least one" mean one or more and include plural referents unless the context clearly dictates otherwise.

[0030] The term "IL-4 (Interleukin-4)," the Japanese name for interleukin-4, is an important cytokine released by Th2 cells, which plays a central role in asthma and bronchial allergic inflammatory responses and is involved in the production of IgE immunoglobulins.

[0031] The term "IL-13 (Interleukin-13)," the Japanese name for interleukin-13, is an important cytokine released by Th2 cells that plays a central role in asthma and bronchial allergic inflammatory responses and can promote remodeling of the airway epithelium, mucus secretion, and hyperresponsiveness of bronchial smooth muscle.

[0032] In order to explain the technical features and effects of the present invention, several examples will be given below and will be described in detail with reference to the drawings.

[0033] The dosage of the composition disclosed in the animal experiments in the following examples varies depending on factors such as the subject of administration, the dosage form of the composition, and the composition form, and the dosage change can be calculated by a person skilled in the art using common sense. For example, the dosage of the composition of the present invention when administered to mice is 45.05 mg / kg, which corresponds to a human dosage of 3.65 mg / kg (about 219 mg / day) for an adult weighing 60 kg. [Example]

[0034] Example 1: Preparation of Polypeptides The polypeptides of SEQ ID NOS: 1 to 10 shown in Table 1 above were prepared by artificial synthesis, and the amino acid sequence of each polypeptide was confirmed to be correct.

[0035] Example 2: Cellular testing In this example, the P815 rat mast cell line was used as the cell line, and Compound 48 / 80 was used as the allergen. First, each of the polypeptides shown in SEQ ID NOS: 1 to 10 of the present invention was co-cultured with P815 rat mast cells for 20 minutes, followed by the addition of Compound 48 / 80 and co-culture for 6 hours. After completion of the culture, 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.

[0036] 1 and Table 2, it was found that the polypeptides of the present invention shown in SEQ ID NOs: 1 to 10 each have IL-4 suppressive activity, and the polypeptides of SEQ ID NOs: 4, 5, and 9 each have IL-13 suppressive activity. Overall, it was found that the polypeptides of SEQ ID NOs: 4, 6, 8, and 9 have excellent Th2 pathway suppressive activity.

[0037] It was found that the polypeptides of the present invention shown in SEQ ID NOS: 1 to 10 each suppress the increase in IL-4 and / or IL-13 caused by allergic reactions, reduce the increase in allergic phenomena leaning toward the Th2 ratio, and are effective in improving inhalation allergic reactions, such as diseases including bronchial hypersensitivity, asthma, bronchitis, etc. In other words, administering an effective amount of any of the polypeptides of the present invention shown in SEQ ID NOS: 1 to 10 to an individual suffering from bronchial hypersensitivity or a related disease effectively improves or alleviates the symptoms of bronchial hypersensitivity, and the improving effect was superior when the polypeptide shown in SEQ ID NOS: 1, 6, 8, or 9 was administered.

[0038] [Table 2]

[0039] Example 3: Animal Experiments The mice were randomly divided into three groups, and the experimental period was 23 days in total. The treatment conditions for the mice in each group were as follows:

[0040] Group 1 mice (blank group): Normal mice that were not subjected to a sensitization treatment and were not administered with the peptide composition of the present invention.

[0041] The second group of mice (asthma group): in the asthma model, they were challenged with venom from the 16th to the 22nd day of the experiment.

[0042] Group 3 mice (treatment group): These were asthma model mice that were challenged with venom from the 16th to 22nd day of the experiment, and were administered the peptide composition of the present invention every day during the experiment (dosage: 45.05 mg / kg BW, BW is body weight).

[0043] Preparation of asthma model animals: On days 1, 8, and 15 of the experiment, OVA was administered to each mouse to sensitize it and induce airway inflammation in each mouse.

[0044] Toxic challenge treatment: PM2.5 was administered to mice (15 μg / mouse) daily from day 16 to day 22 of the experiment. The PM2.5 samples were standard suspended particulate matter collected from urban environments.

[0045] The active ingredients of the peptide composition of the present invention include the polypeptides shown in SEQ ID NOS: 1 to 10, and each polypeptide is blended in equal or approximately equal amounts.

[0046] After the experiment, the airway resistance (Penh) values ​​of the mice in each group 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 the mice in each group, and their proliferation activity and natural killer cell (NK cell) activity were measured. The results are shown in Figures 4 and 5. Bronchoalveolar lavage fluid (BALF) was collected from the mice in each group, and the concentrations of IL-13, IL-4, and INF-γ in the BALF were measured by ELISA. The results are shown in Figures 6 to 8. In addition, the IgE and IgG1 concentrations in the serum of the mice in each group were evaluated. The results are shown in Figures 9 and 10.

[0047] As can be seen from the results in Figures 2 and 3, the airway resistance of the mice in Group 3 was significantly lower than that of the mice in Group 2, with a reduction of more than 30%, indicating that administration of the peptide composition of the present invention can effectively improve asthma symptoms, i.e., alleviate asthma symptoms.

[0048] As can be seen from the results in Figures 4 and 5, the proliferation activity of spleen cells and the activity of natural killer cells in Group 3 mice were increased compared to Group 2 mice, indicating that administration of the peptide composition of the present invention can suppress excessive activation of spleen cells and restore the balance between Th1 and Th2. In other words, the results in Figures 4 and 5 indicate that the peptide composition of the present invention is useful for improving allergic asthma, particularly overactivated Th2-type asthma.

[0049] As can be seen from the results in Figures 6 to 8, the IL-4 and IL-13 concentrations in the BALF of Group 3 mice were 37.6% and 15.8% lower, respectively, than those of Group 2 mice, and the INF-γ concentration in the BALF of Group 3 mice was 54.5% higher than those of Group 2 mice. As can be seen from the results in Figures 9 and 10, the IgE and IgG1 concentrations in the serum of Group 3 mice were 79.2% and 34.1% lower than those of Group 2 mice. The results in Figures 6 to 10 again demonstrate that the peptide composition of the present invention can actually regulate the expression of bronchial inflammatory cytokines, reduce the concentrations of IL-4 and IL-13, increase the INF-γ concentration, and significantly reduce the IgE concentration in serum, demonstrating that the peptide composition of the present invention can actually prevent or reduce the occurrence of allergic asthma and certainly has the effect of preventing immune imbalance.

[0050] As can be seen from the above explanation, the polypeptides of the present invention shown in SEQ ID NOs: 1 to 10 each have IL-4 and / or IL13 suppressive activity, and therefore, by administering an effective amount of the polypeptide of the present invention or a composition containing the polypeptide to an individual with bronchial allergy symptoms or an individual at high risk, diseases induced by bronchial hypersensitivity, such as asthma, can be prevented or ameliorated.

Claims

1. A polypeptide having an amino acid sequence represented by any one of SEQ ID NOs: 1 to 10.

2. A peptide composition for suppressing IL-4 and / or IL13, comprising at least one polypeptide, the amino acid sequence of which is represented by any one of SEQ ID NOs: 1 to 10.

3. A peptide composition for treating or preventing a disease associated with bronchial hyperresponsiveness, comprising at least one polypeptide, the amino acid sequence of which is represented by any one of SEQ ID NOs: 1 to 10.

4. A peptide composition for treating or preventing bronchial hyperresponsiveness-related diseases according to claim 2, wherein the peptide composition comprises polypeptides shown in SEQ ID NO: 4, SEQ ID NO: 6, SEQ ID NO: 8 and SEQ ID NO:

9.

5. A peptide composition for treating or preventing bronchial hyperresponsiveness-related diseases as described in claim 3, wherein the peptide composition comprises polypeptides shown in 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.

6. The peptide composition for treating or preventing a disease associated with bronchial hyperresponsiveness according to claim 2, wherein the peptide composition is used to suppress the activity of IL-4 and / or IL-13.

7. The peptide composition for treating or preventing bronchial hyperresponsiveness-related diseases according to claim 2, wherein the peptide composition is a food, a nutritional supplement, or a pharmaceutical.

8. The peptide composition for treating or preventing bronchial hyperresponsiveness-related diseases according to claim 2, wherein the peptide composition is used to reduce the concentration of IgE.

9. The peptide composition for treating or preventing a disease associated with bronchial hyperresponsiveness according to claim 2, wherein the disease associated with bronchial hyperresponsiveness is asthma.

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