KR102508935B1 - Anti-allergy composition comprising velutin - Google Patents

Abstract

The present invention relates to a composition containing velutin for preventing, treating or improving allergic diseases. As such, the composition containing velutin of the present invention can be usefully used as an antihistamine composition with improved side effects in the cosmetic, food, or pharmaceutical fields where an anti-allergic composition is required.

Description

Anti-allergy composition comprising velutin {Anti-allergy composition comprising velutin}

The present invention relates to a composition for preventing, treating or improving allergic diseases, including Velutin.

Flavonoids are a type of phenolic compounds that exist in nature, and various physiological activities such as antioxidant, anti-inflammatory, antibacterial, and anticancer have been reported. Due to the various physiological activities of flavonoids, they are widely used as health functional foods, functional cosmetics, and medicinal materials. In particular, structural diversity is possible depending on the position of the hydroxyl group and the methoxy group of the flavonoid, and it exhibits various physiological activities.

Velutin is a flavonoid found in natural products such as Viscum album, Flammulina velutipes, Acai berry (Euterpe oleracea Mart), and Suaeda japonica. It has been reported to exhibit various physiological activities such as inhibition and strong bitter taste masking effect.

On the other hand, allergy is an overreaction of the immune system caused by a foreign antigen (allergen) in an innocuous environment. Symptoms such as itching, fever, and pain, which are the main symptoms of asthma, atopic dermatitis, and allergic rhinitis, appear through an allergic reaction, and these symptoms are caused by excessive activation of mast cells.

Mast cells play a critical role in the initiation of allergic reactions, and activation of mast cells is closely related to specific immunoglobulin E (IgE) antibodies as typical activators. IgE binds to Fcε receptor I (Fc epsilon receptor I, FcεRI) expressed on the surface of mast cells. When FcεRI cross-links with an antigen on mast cells, an intracellular signaling cascade is induced, and Lyn, Syk, protein kinase C (PKC), and mitogen activated protein kinase (mitogen activated protein kinase, Downstream mediators such as MAPK) are activated and intracellular Ca2+ (calcium ion) concentration increases. In addition, the aggregation of IgE-FcεRI induces exocytosis of preformed granules containing histamine, chemokines, cytokines, and serine proteases, and leukotrienes and prostaglandins stimulates the new synthesis of These inflammatory mediators cause a variety of pathophysiological symptoms in the allergic response, including contraction of airway smooth muscle, secretion of mucus, and recruitment of other inflammatory cells. Thus, regulation of inflammatory mediator production is important for controlling allergic responses.

Rat basophilic leukemia (RBL)-2H3 cells are rodent mucosal mast cells secreting inflammatory mediators. Although the RBL-2H3 cell line does not completely represent primary mast cells and basophils, the RBL-2H3 cell line shares the characteristics of the two cells and allows for simple subculture. A key feature of mast cells is the secretion of histamine. Histamine is involved in allergic and inflammatory responses and plays an important role in mediating interactions between different cells. Also, histamine is secreted from RBL-2H3 cells. Therefore, RBL-2H3 cells have generally been used for in vitro studies of allergic reactions. Phobol myristate acetate (PMA), a PKC activator, and A23187, a calcium ionophore, increase intracellular calcium ion concentration and induce activation of signaling pathways in RBL-2H3 cells do.

Under various reports of these allergy-related mechanisms, many studies are being conducted to develop more effective anti-allergic substances.

Korean Patent Publication No. 10-2004-0087223 Korean Patent Publication No. 10-2018-0052226

The inventors of the present invention recognized the side effects of conventional antihistamines, such as drowsiness, dizziness, vomiting, diarrhea, indigestion, excitement, etc. when taken, and while studying a substance for which these side effects were alleviated, velutene was found to be released from mast cells. It was confirmed that the degranulation was inhibited by inhibiting the release of histamine and β-hexosaminidase.

Accordingly, an object of the present invention is to provide a pharmaceutical composition for preventing or treating allergic diseases, including Velutin.

Another object of the present invention is to provide a food composition for preventing or improving allergic diseases, including velutin.

Another object of the present invention is to provide a cosmetic composition for preventing or improving allergic diseases, including velutin.

In order to achieve the object of the present invention, the present invention provides a pharmaceutical composition for preventing or treating allergic diseases, including velutin.

In addition, the present invention provides a food composition for preventing or improving allergic diseases, including velutin.

In addition, the present invention provides a cosmetic composition for preventing or improving allergic diseases, including veloutin.

In one embodiment of the present invention, the veloutine may have a therapeutic concentration of 1 to 200 μg/ml.

In one embodiment of the present invention, the allergic disease may be an allergic inflammatory disease.

In one embodiment of the present invention, the allergic disease may be one selected from the group consisting of asthma, hay fever (allergic rhinitis) and atopic eczema due to an immediate allergic reaction.

In one embodiment of the present invention, the composition can inhibit degranulation of mast cells.

In one embodiment of the present invention, the composition can inhibit β-hexosaminidase or histamine release.

According to the present invention, the composition containing velutin inhibits the release of β-hexosaminidase and histamine from mast cells and exhibits the effect of inhibiting degranulation, thereby reducing drowsiness, dizziness, vomiting, diarrhea, indigestion, excitement, etc. It has been found that it can mitigate the side effects of antihistamines. Therefore, the composition containing velutin of the present invention can be usefully used as an antihistamine composition with improved side effects in the field of cosmetics or pharmaceuticals requiring an anti-allergic composition.

1 shows the structures of velutein and similar compounds of the present invention, Genkwanin and luteolin.
Figure 2 shows the excellent β-hexosaminidase and histamine release inhibitory effect upon treatment with velutin of the present invention.
3 shows the ¹ H-NMR spectrum of the prepared velutin.
Figure 4 shows the ¹ H-NMR spectrum of the prepared genkwanin.

The present invention provides a pharmaceutical composition for preventing or treating allergic diseases, including Velutin.

In the present invention, the velutinin may have a therapeutic concentration of 1 to 200 μg/ml, preferably 50 to 150 μg/ml, and most preferably 100 μg/ml, but is not limited thereto.

In one embodiment of the present invention, the allergic disease may be an allergic inflammatory disease, preferably an immediate allergic reaction, more preferably an immediate allergic reaction such as asthma, hay fever (allergic rhinitis) ) and atopic eczema.

The term allergic disease used in the present invention means a disease caused by allergy. Allergy (allergy) refers to an antigen-antibody reaction that occurs when an antibody is made against a certain type of substance (antigen or allergen) when it enters the body, and then the same substance antigen enters the body again. The main diseases are bronchial asthma, allergic rhinitis, hives, drug rash, drug allergy, serum sickness, etc., and various types of diseases are represented depending on the type of allergen or the tissue causing the allergic reaction. In addition, in the present invention, an allergic inflammatory disease means an inflammatory disease induced by the allergic reaction.

The term immediate-type allergic reaction used in the present invention refers to type I among four types of allergic reactions: type I, type II, type III, and type IV. In the type I reaction, when IgE binds to the surface of mast cells or basophils and a specific antigen binds to the IgE antibody, granule escape (degranulation) occurs in mast cells or basophils, resulting in histamine and serotonin It refers to a phenomenon in which chemical substances such as bradykinin are released, resulting in enhancement of vascular permeability and promotion of mucus secretion. Symptoms included in this include anaphylaxis, bronchial asthma, allergic rhinitis, and urticaria.

The composition of the present invention can inhibit degranulation of mast cells and inhibit the release of β-hexosaminidase or histamine. Mast cells cause hypersensitivity reactions (allergies) when activated, and these hypersensitivity reactions cause diseases such as atopy and asthma. Mast cells have numerous intracellular granules, and the granules contain a number of inflammatory substances such as histamine and TNF-α. When mast cells are activated, these granules fuse with the cell membrane, and substances in the granules are secreted to the outside of the cell, triggering an overactive inflammatory response. This process is called degranulation. β-hexosaminidase is a substance secreted simultaneously with histamine when mast cell degranulation occurs, and is widely used as an indicator to determine the degree of mast cell degranulation.

As used herein, the term "prevention" refers to any action that suppresses or delays the onset of allergic diseases by administration of the pharmaceutical composition according to the present invention.

As used herein, the term "treatment" refers to all actions that improve or beneficially change symptoms caused by allergic diseases by administration of the pharmaceutical composition according to the present invention.

The pharmaceutical composition according to the present invention includes velutin as an active ingredient and may include a pharmaceutically acceptable carrier. The pharmaceutically acceptable carrier is one commonly used in formulation and includes, but is not limited to, saline solution, sterile water, Ringer's solution, buffered saline solution, cyclodextrin, dextrose solution, maltodextrin solution, glycerol, ethanol, liposome, etc. It is not, and if necessary, other conventional additives such as antioxidants and buffers may be further included. In addition, diluents, dispersants, surfactants, binders, lubricants, and the like may be additionally added to prepare formulations for injections such as aqueous solutions, suspensions, emulsions, etc., pills, capsules, granules, or tablets. Regarding a suitable pharmaceutically acceptable carrier and formulation, it can be preferably formulated according to each component using the method disclosed in Remington's literature. The pharmaceutical composition of the present invention is not particularly limited in dosage form, but may be formulated as an injection or oral intake.

The pharmaceutical composition of the present invention can be administered orally or parenterally (for example, intravenously or subcutaneously) depending on the desired method, and the dosage is the condition and weight of the patient, the severity of the disease, the form of the drug, Depending on the route and time of administration, it can be appropriately selected by those skilled in the art.

The composition according to the present invention is administered in a pharmaceutically effective amount. In the present invention, "pharmaceutically effective amount" means an amount sufficient to treat a disease at a reasonable benefit / risk ratio applicable to medical treatment, and the effective dose level is based on the type, severity, and activity of the drug in the patient. , sensitivity to the drug, time of administration, route of administration and excretion rate, duration of treatment, factors including drugs used concurrently, and other factors well known in the medical field. The composition according to the present invention may be administered as an individual therapeutic agent or in combination with other therapeutic agents, may be administered sequentially or simultaneously with conventional therapeutic agents, and may be administered single or multiple times. Considering all of the above factors, it is important to administer an amount that can obtain the maximum effect with the minimum amount without side effects, which can be easily determined by those skilled in the art.

Specifically, the effective amount of the composition according to the present invention may vary depending on the patient's age, sex, and weight, and may increase or decrease depending on the route of administration, severity of disease, gender, weight, age, and the like.

As used herein, the term "pharmaceutically acceptable salt thereof" can be prepared by a conventional method in the art, for example, hydrochloric acid, hydrogen bromide, sulfuric acid, sodium hydrogen sulfate, phosphoric acid, carbonic acid Salts with inorganic acids such as, or together with organic acids such as formic acid, acetic acid, oxalic acid, benzoic acid, citric acid, tartaric acid, gluconic acid, gestisic acid, fumaric acid, lactobionic acid, salicylic acid, or acetylsalicylic acid (aspirin) Forming chemically acceptable acid salts thereof, or reacting with alkali metal ions such as sodium and potassium to form metal salts thereof, or reacting with ammonium ions to obtain another form of pharmaceutically acceptable salt thereof. means to form

As another aspect, the present invention provides a food composition for preventing or improving allergic diseases, including veloutin.

The food composition is a concept including health functional food.

As used herein, the term "improvement" refers to all actions that at least reduce the parameters related to the condition to be treated, for example, the severity of symptoms.

In the food composition of the present invention, veloutine may be added to food as it is or used together with other foods or food ingredients, and may be appropriately used according to conventional methods. The mixing amount of velutin can be suitably determined according to its purpose of use (for prevention or improvement). In general, when preparing food or beverage, the composition of the present invention is added in an amount of 15% by weight or less, preferably 10% by weight or less, based on the raw material. However, in the case of long-term intake for the purpose of health and hygiene or health control, the amount may be less than the above range.

The health functional food composition of the present invention has no particular restrictions on other ingredients other than containing the above ingredients as essential ingredients in the indicated proportions, and may contain various flavoring agents or natural carbohydrates as additional ingredients like conventional beverages. . Examples of the aforementioned natural carbohydrates include monosaccharides such as glucose, fructose, and the like; disaccharides such as maltose, sucrose and the like; and polysaccharides such as conventional sugars such as dextrins, cyclodextrins, and the like, and sugar alcohols such as xylitol, sorbitol, and erythritol. As flavoring agents other than those mentioned above, natural flavoring agents (thaumatin, stevia extract (eg rebaudioside A, glycyrrhizin, etc.) and synthetic flavoring agents (saccharin, aspartame, etc.) can advantageously be used. The ratio of the natural carbohydrates can be appropriately determined by a person skilled in the art.

In addition to the above, the health functional food composition of the present invention includes various nutrients, vitamins, minerals (electrolytes), flavors such as synthetic flavors and natural flavors, colorants and enhancers (cheese, chocolate, etc.), pectic acid and its salts, Alginic acid and salts thereof, organic acids, protective colloidal thickeners, pH adjusting agents, stabilizers, preservatives, glycerin, alcohol, carbonating agents used in carbonated beverages, and the like may be contained. These components may be used independently or in combination. The ratio of these additives can also be appropriately selected by those skilled in the art.

As another aspect, the present invention provides a cosmetic composition for preventing or improving allergic diseases, including veloutine.

In the present invention, the cosmetic composition is a formulation selected from the group consisting of solutions, suspensions, emulsions, pastes, gels, creams, powders, soaps, surfactant-containing cleansing, oils, powder foundations, emulsion foundations, wax foundations and sprays. It may have, but is not limited thereto.

The cosmetic composition of the present invention may include components commonly used in cosmetic compositions, for example, antioxidants, stabilizers, solubilizers, vitamins, pigments, and conventional auxiliary agents such as fragrances, and carriers. .

In addition, the composition of the present invention may be used in combination with an organic sunscreen agent. The organic sunscreens include glyceryl parba, drometrizoletrisiloxane, drometrizole, digaloyltrioleate, disodium phenyldibenzimidazole tetrasulfonate, diethylhexylbutamidotriazone, diethylamino Hydroxybenzoylhexylbenzoate, DEA-methoxycinnamate, mixture of Lawson and dihydroxyacetone, methylenebis-benzotriazolyltetramethylbutylphenol, 4-methylbenzylidenecamphor, menthylantranilate, benzophenone -3 (oxybenzone), benzophenone-4, benzophenone-8 (dioxyphebenzone), butylmethoxydibenzoylmethane, bisethylhexyloxyphenol methoxyphenyltriazine, cinoxate, ethyldihydroxypropylparba, Octocrylene, Ethylhexyldimethylparba, Ethylhexylmethoxycinnamate, Ethylhexyl salicylate, Ethylhexyltriazone, Isoamyl-p-methoxycinnamate, Polysilicone-15 (dimethoxydiethylbenzalmalonate) , At least one selected from the group consisting of terephthalylidene dicamphor sulfonic acid and its salts, TEA-salicylate, and aminobenzoic acid (Pava) may be used.

Hereinafter, the present invention will be described in more detail through examples. However, the following examples are intended to illustrate the present invention, and the scope of the present invention is not limited thereto.

<Example 1: Synthesis of veloutine and genkwanin>

Among the three flavone derivatives, velutein and genkwanin were chemically synthesized, and luteolin was purchased from ChemOne Co., Ltd. (ChemOne, Korea).

Production of veloutine and genkwanin: It was prepared from 2',4',6'-trihydroxyacetophenone through 4 sequential methylation, aldol condensation, cyclization, and deprotection reactions (Fig. 3 and see 4).

Preparation of velutin: ¹ H NMR (DMSO-d ₆ , 400 MHz), δ : 12.97 (s, 1H), 10.00 (s, 1H), 7.61-7.59 (m, 2H, J = 4.62 Hz), 6.96-6.93 (m, 2H, J = 9.96 Hz), 6.80 (d, 1H, J = 2.24 Hz), 6.38-6.37 (d, 1H, J = 2.20 Hz), 3.93 (s, 3H), 3.91 (s, 3H) .

Preparation of genkwanin: ¹ H NMR (DMSO-d ₆ , 400 MHz), δ : 12.96 (s, 1H), 7.96 (d, 2H, J = 8.84 Hz), 6.93 (d, 2H, J = 8.84 Hz) , 6.84 (s, 1H), 6.77 (d, 1H, J = 2.25 Hz), 6.37 (d, 1H, J = 2.25 Hz), 3.87 (s, 1H)

<Example 2: Experiment preparation>

RBL-2H3 cells were grown in DMEM medium (Dulbecco's modified Eagle's medium, Hyclone Logan, UT, USA) supplemented with 10% heat-inactivated fetal bovine serum (FBS), 1% penicillin, and 1% streptomycin (Hyclone). cultured.

<Example 3: Checking the effect of velutin on reducing β-hexosaminidase release>

RBL-2H3 (mast cell) cells were seeded in a 24-well plate using DMEM media containing 10% FBS and cultured for 24 hours. Then, the culture medium was removed and velutin was added to siraganian buffer (NaCl 119mM, KCl 5mM, Glucose 5.6mM, PIPES 25mM, MgCl ₂ , 0.4mM, CaCl ₂ 1mM, BSA 0.1%, pH to a concentration of 2-100 μg/ml). 7.2) and treated the cells for 1 hour. Genkwanin and luteolin were also treated at 100 μg/ml concentration as a control to compare the effect of velutin. Then, the solution was removed from the cells, washed twice using PIPES, and an immune response was induced by treatment with 50 nM PMA (phorbol myristate acetate) and 1 μM A23187 (calcium ionophore) for 30 minutes. Then, 20 μL of the supernatant was transferred to a 96-well plate, and the substrate, 4-p-nitrophenyl-N-acetyl-β-D-glucosaminide, was diluted in 0.05 M sodium citrate buffer (pH 4.5) to a concentration of 2 mM. It was mixed with 80 μL and reacted at 37 degrees for 30 minutes. Thereafter, 200 μL of stop buffer (0.1 M·NaHCO ₃ , pH 10) was added to terminate the reaction, and absorbance was measured at 405 nm.

As a result, as shown in FIG. 2, β-hexosaminidase release was significantly increased when degranulation was activated compared to normal mast cells (-) that were not treated with anything, and β-hexosaminidase release decreased as the concentration of verrutin treatment increased. Confirmed. On the other hand, it was confirmed that genkwanin and luteolin showed lower effects than velutin even when treated at the same concentration of 100 μg/ml.

<Example 4: Checking the histamine release reducing effect of velutin>

RBL-2H3 (mast cell) cells were seeded in a 24-well plate using DMEM media containing 10% FBS and cultured for 24 hours. Then, the culture medium was removed and velutin was added to siraganian buffer (NaCl 119mM, KCl 5mM, Glucose 5.6mM, PIPES 25mM, MgCl ₂ , 0.4mM, CaCl ₂ 1mM, BSA 0.1%, pH to a concentration of 2-100 μg/ml). 7.2) and treated the cells for 1 hour. Genkwanin and luteolin were also treated at 100 μg/ml concentration as a control to compare the effect of velutin. Then, the solution was removed from the cells, washed twice with PIPES, and treated with 50 nM PMA (phorbol myristate acetate) and 1 μM A23187 (calcium ionophore) for 30 minutes to induce an immune response. Then, 1 mL of the supernatant was mixed with 2 mL of 1 N NaOH and 0.1 mL of 1% OPT (o-phtha-laldehyde) and reacted at room temperature for 5 minutes. Then, the reaction was terminated by the addition of 0.1 mL 3 N·HCl, and fluorescence intensity excitation at 360 nm and emission at 450 nm was measured.

As a result, as shown in FIG. 2, it was confirmed that histamine release was significantly increased when degranulation was activated compared to normal cells (-), which were not treated with anything, and histamine release decreased as the concentration of berutin increased. On the other hand, it was confirmed that genkwanin and luteolin showed lower effects than velutin even when treated at the same concentration of 100 μg/ml.

Claims (8)

An antihistamine pharmaceutical composition for preventing or treating allergic diseases, comprising velutin as an active ingredient, and suppressing histamine release from mast cells to inhibit degranulation of mast cells.
The antihistamine pharmaceutical composition according to claim 1, wherein the velutinin has a therapeutic concentration of 1 to 200 μg/ml.
delete delete delete delete An antihistamine food composition for preventing or improving allergic diseases, comprising velutin as an active ingredient, and suppressing histamine release from mast cells to inhibit degranulation of mast cells.
An antihistamine cosmetic composition for preventing or improving allergic diseases, comprising velutin as an active ingredient, and suppressing histamine release from mast cells to inhibit degranulation of mast cells.

Images