WO2020130166A1 - Composition comprising magnolia flos extract for preventing and treating allergic rhinitis - Google Patents

Abstract

The present invention provides an inhalation aerosol preparation comprising a Magnolia flos extract, which can be effectively used for preventing and treating allergic rhinitis. The present invention provides an aerosol preparation comprising a particular concentration of a Magnolia flos extract, which is parenterally administered by inhalation and as such can be advantageously utilized as a composition for prevention and treatment of allergic rhinitis disease.

Description

A composition for the prevention and treatment of allergic rhinitis comprising an extract of Shin

The present invention relates to an aerosol formulation for inhalation containing a kidney extract that can be effectively used for preventing and treating allergic rhinitis.

Allergic rhinitis is a chronic inflammatory disease characterized by a continuous sneezing seizure, a continuous runny nose, and nasal congestion, in which the nasal mucosa is hypersensitive to certain substances (allergens). When allergens such as pollen and house dust mites are exposed to the nasal mucosa through breathing, inflammatory cells recognize this and activate Th2 T cells, mast cells, and the like. Among these, Th2 T cells secrete cytokines such as IL-4 and IL-13, causing class switching, in which B cells secrete IgE, which binds to the Fc receptor of mast cells and mediates IgE. Initiate an allergic reaction. Mast cells with specific IgE binding to specific allergens are degranulated to secrete substances such as histamine that are activated and stored by allergens. The secretion of histamine is combined with histamine receptors expressed in nasal mucosal epithelial cells and blood vessels, causing hypersecretion of mucus, stuffy nose, and the like.

On the basis of this mechanism, the treatment of inflammatory diseases including allergic rhinitis is generally used as an immunosuppressive agent and an antihistamine agent for suppressing the activity of Th2 T cells and mast cells, which can primarily cause allergies.

The most potent immunosuppressant present in the Limousine family of drugs (tacrolimus, sirolimus, etc.) inhibits the activity of a transcription regulator called NFAT (Nuclear factor of activated T-Cells) in immune cells, resulting in inhibition of pro-inflammatory cytokine production. It is known that NFAT is activated by a protein phosphatase called calcineurin.

In the case of an antihistamine, it specifically binds to the histamine receptor expressed on mast cells, and suppresses degranulation so that mast cells cannot be activated and secrete histamine even when exposed to allergens. However, these effects not only require the use of two drugs, such as anti-inflammatory and anti-histamine, all at once, but also increase the risk of cancer (inflammation inhibitor) and sedation (antihistamine), drowsiness helplessness, and learning ability may decrease. It can cause mental and physical discomfort.

According to a recent study, it is known that intracellular calcium signal generation is important for activation of immune cells and mast cells, and it is known that these pathways that cause intracellular calcium increase are shared with each other.

In the case of stimulation through the T cell receptor or stimulation via the FC receptor, Gq protein-linked receptors are activated to activate PLC (phospholipase C).The activity of these PLCs is IP ₃ (inositol-3-phosphate) and DAG (diacylglycerol). Will be created.

They ultimately deplete the ER calcium reservoir, which activates the Store Operated Calcium Entry (SOCE), an extracellular calcium entry pathway common to these cells. In addition, according to other recent findings, the TRPV1 ion pathway, which was thought to be limited to neurons that cause pain (DRG neuron), is also present in Th2 immune cells and plays a pivotal role in activating immune cells along with SOCE. Turned out.

On the other hand, this increase in intracellular calcium concentration occurs in nasal epithelial cells, which are important for runny nose formation when allergen is exposed. When the intracellular calcium concentration increases, it activates a calcium-dependent chlorine ion pathway called ANO1 in response to the increased intracellular calcium ion. ANO1 activation accelerates the production of runny nose, causing the secretion of a clear runny nose that is a characteristic symptom of allergic rhinitis.

Therefore, the present inventor simultaneously inhibits SOCE, TRPV1 and ANO1, which is known to be useful for the prevention and treatment of allergic rhinitis, and inhales for effective allergic rhinitis treatment that can replace the administration of immunosuppressants and antihistamines without side effects. The present invention has been completed by developing an aerosol formulation for use.

One object of the present invention is to provide an aerosol formulation for inhalation containing a kidney extract that can be effectively used for preventing and treating allergic rhinitis.

One aspect of the present invention provides an aerosol formulation for inhalation for the prevention and treatment of allergic rhinitis containing 0.001% to 0.08% by weight of the Sini extract.

According to one embodiment of the present invention, the aerosol formulation for inhalation may be one containing 0.01% by weight to 0.08% by weight of the extract.

The present invention can be usefully used as a composition for the prevention and treatment of allergic rhinitis diseases, by providing an aerosol preparation for inhalation, which is a parenteral administration method containing a Shini extract in a specific concentration.

1 is a graph showing the inhibitory effect of calcium ion current (I _SOCE ) through ORAI1 of 30% ethanol extract of herbal medicine according to an embodiment of the present invention, ORAI1 ion measured in HEK293-T cells overexpressed with ORAI1 ion channel It shows the patch clamp chart records of 30% ethanol extract 1 mg/ml (red arrow) and ORAI1 inhibitor BTP2 (blue arrow) showing the current of the passage (black arrow) and its inhibitory effect.

FIG. 2 is a graph showing a current-voltage (I-V) relationship curve of the corresponding arrow shown in FIG. 1.

3 is a graph summarizing the degree of inhibition of the current measured at the 120 mV voltage of the recorded I _SOCE current.

FIG. 4 is a graph showing the inhibitory effect of TRPV1 current, which is a non-selective cation channel of 30% ethanol extract, and the current (black arrow) of TRPV1 ion channel measured in HEK293-T cells overexpressing TRPV1 ion channel and its inhibition Shows the patch clamp chart record of 30 μg/ml (red arrow), 30 μg/ml (blue arrow), 100 μg/ml (blue green arrow) and TRPV1 inhibitor BCTC (gray arrow) of 30% ethanol extract showing the effect.

5 is a graph showing a current-voltage (I-V) relationship curve of the corresponding arrow shown in FIG. 4.

FIG. 6 shows the suppression of T cell proliferation by separating immune cells T cells from PBMC collected from human blood and stimulating them with CD3 and CD28, 30% ethanol extract showing the inhibitory effect of ORAI1 ion current 0.1 It shows the percentage of T cell proliferation inhibition and cell proliferation ratio of mg/ml, 0.3 mg/ml, and 1 mg/ml. Negative refers to T cells that did not stimulate, and Positive refers to cells that stimulated T cells with CD3 and CD28 to induce proliferation.

FIG. 7 is a graph showing the inhibitory effect of ANO1 current, which is a calcium-dependent chloride ion channel of 30% ethanol extract, and the current of ANO1 ion channel and HE 30% ethanol extract 0.1 measured in HEK293-T cells overexpressing ANO1 ion channel. Patch clamp chart records of A0-1, mg/ml, 0.3 mg/ml, 1 mg/ml and ANO1 inhibitors are shown.

8 is a graph showing the current voltage (I-V) relationship curve of FIG. 7.

9 is a graph summarizing the degree of inhibition of the current measured at a voltage of +100 mV of the recorded ANO1 current.

10 is a graph showing the current inhibitory effect of the calcium-dependent chloride ion channel (I _CaCC ) measured in human airway epithelial cell line Calu-3, an allergen-induced cytokine secreted by airway epithelial cells Calu-3 and Th2 T cells. Compared by measuring the increased calcium-dependent chloride ion channel current in Calu-3 treated with IL-4 100 ng/ml for 24 hours and the current and ANO1 measured at +100 mV voltage of the recorded calcium-dependent chloride ion channel current It shows the current suppression of the ion channel inhibitor Ani9.

11 is a graph showing the current inhibitory effect of the calcium-dependent chloride ion channel (I _CaCC ) measured in the human airway epithelial cell line Calu-3, 30% ethanol extract from Shin in Calu-3 cells treated with cytokine IL-4 0.1 mg/ml, 0.3 mg/ml, 1 mg/ml of calcium-dependent chloride ion channel current suppression and ANO1 ion channel inhibitor Ani9 current suppression and recorded calcium-dependent chloride ion channel current measured at + 100 mV voltage It shows a summary of the degree of inhibition of the current.

FIG. 12 is a graph showing the therapeutic effect of 30% ethanol extract from Shin in an animal model of allergic rhinitis-induced disease in mice. It shows the average score of clinical symptoms, which is the sum of the sneeze and nose rubbing observed for a minute.

FIG. 13 is a graph showing the therapeutic effect of 30% ethanol extract from kidney in animal models of allergic rhinitis-induced disease in mice. Cyto secreted from spleen cells collected from animal models of kidney treated by concentration of 30% ethanol extract Cain IL-4.

14 is a graph showing the therapeutic effect of 30% ethanol extract from kidney in the animal model of allergic rhinitis-induced disease in mice, and the cytokine secreted from spleen cells collected from the animal model in which the kidney was treated by concentration of 30% ethanol extract Cain IL-13.

In order to achieve the above object, the present invention provides an aerosol formulation for inhalation for the prevention and treatment of allergic rhinitis, which contains 0.001 to 0.08% by weight of the Sini extract.

The present inventors studied the pharmacological mechanism of Shini extract, and then inhibited the action of calcium ion current (I _SOCE ) through ORAI1, inhibition of TRPV1 current, which is a non-selective cation pathway, and inhibition of ANO1 current, which is a calcium-dependent chloride ion channel. By this, it was confirmed that the extract has an effect on preventing and treating allergic rhinitis.

Preventing and treating allergic rhinitis by the inhibitory effect of the extract of Shini according to the present invention through the inhibition of calcium ion current (I _SOCE ) through ORAI1, inhibition of TRPV1 current, which is a non-selective cation pathway, and ANO1 current, which is a calcium-dependent chloride ion channel. It is shown in Figures 1 to 11, Examples 2 to 6 showing an effect on the.

The present invention is the most effective at concentrations containing 0.001% by weight (0.01 mg/ml) to 0.08% by weight (0.8 mg/ml) of Shini extract after studying several pharmacological mechanisms for preventing and treating allergic rhinitis of the Sini extract. Was confirmed.

Therefore, the aerosol formulation for inhalation used in the present invention contains 0.001 to 0.08% by weight of the extract of Shini, and preferably 0.01 to 0.08% by weight. If the extract of Shini is administered at a concentration of less than 0.001% by weight, it is difficult to expect a preventive and therapeutic effect against allergic rhinitis, and surprisingly, even if the Sini extract is administered at a concentration of 0.08% by weight (0.8 mg/ml) or more, It has no preventive and therapeutic effect.

Preventing and treating allergic rhinitis by the inhibitory effect of the extract of Shini according to the present invention through the inhibition of calcium ion current (I _SOCE ) through ORAI1, inhibition of TRPV1 current, which is a non-selective cation pathway, and ANO1 current, which is a calcium-dependent chloride ion channel. It is shown in Figures 1 to 11, Examples 2 to 6 showing an effect on the.

The present invention is the most effective at concentrations containing 0.0001% by weight (0.001 mg/ml) to 0.008% by weight (0.08mg/ml) of Shini extract after studying several pharmacological mechanisms for preventing and treating allergic rhinitis of the Sini extract Was confirmed.

Therefore, the aerosol formulation for inhalation used in the present invention contains 0.001 to 0.08% by weight of the extract of Shini, and preferably 0.01 to 0.08% by weight. If the extract of Shini is administered at a concentration of less than 0.001% by weight, it is difficult to expect a preventive and therapeutic effect against allergic rhinitis, and surprisingly, even if the Sini extract is administered at a concentration of 0.08% by weight (0.8 mg/ml) or more, It has no preventive and therapeutic effect.

The present invention can be administered by inhaling the Shini extract contained in the aerosol formulation for inhalation once to several times a day.

Shinyi extract used in the present invention can be prepared by the following method. That is, after removing the foreign matter by washing the water with water, it is dried and crushed in the shade. Shini can be used without restrictions such as those grown or commercially available. An appropriate amount of solvent is added to the powder of the crushed kidney so that it is completely immersed. Shini's mixed powder can be extracted using a conventional extraction solvent, preferably, (a) anhydrous or hydrous lower alcohols having 1 to 4 carbon atoms (eg methanol, ethanol, propanol, butanol, normal-propanol, iso -Propanol and normal-butanol, etc.), (b) a mixed solvent of the lower alcohol and water, (c) acetone, (d) ethyl acetate, (e) chloroform, (f) 1,3-butylene glycol, ( g) Hexane, (h) diethyl ether, (i) butyl acetate or (j) can be extracted with water, and the ethanol solution used as a solvent is extracted with ethanol 30: distilled water 70 in terms of extraction yield. desirable. When extracting the medicine with ethanol, it is preferable to extract by heating with a heavy bath for 3 hours, and ethanol may be added at 5 to 20 parts by weight per 1 part by weight. In addition, the extract may be prepared in powder form by additional processes such as distillation under reduced pressure and freeze drying or spray drying.

According to one embodiment of the present invention, the extract may be extracted with any one or more solvents selected from the group consisting of alcohols having 1 to 4 carbon atoms and mixed solvents thereof.

According to one embodiment of the present invention, the extract may be extracted by using a mixed solvent in which the weight ratio of ethanol and water is 3:10 to 6:10.

According to one embodiment of the present invention, the composition may include as much as 0.001% to 0.08% by weight of the Shini extract.

In addition, the present invention is to provide an aerosol formulation for inhalation containing the extract of the kidney.

The carrier contained in the aerosol formulation for inhalation of the present invention is commonly used in the formulation, lactose, dextrose, sucrose, sorbitol, mannitol, starch, acacia rubber, calcium phosphate, alginate, gelatin, calcium silicate, Microcrystalline cellulose, polyvinylpyrrolidone, cellulose, water, syrup, methyl cellulose, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate and mineral oil, but is not limited thereto. . The pharmaceutical composition of the present invention may further include a lubricant, a wetting agent, a sweetener, a flavoring agent, an emulsifying agent, a suspending agent, a preservative, etc. in addition to the above components. Suitable pharmaceutically acceptable carriers and formulations are described in detail in Remington's Pharmaceutical Sciences (22th ed., 2013).

In addition, the aerosol formulation for inhalation of the present invention may include various bases and/or additives necessary and appropriate for the formulation of the formulation, and non-ionic surfactants, silicone polymers, constitution pigments, and the like within the scope of not impairing the effect. Fragrances, preservatives, fungicides, oxidation stabilizers, organic solvents, ionic or nonionic thickeners, softening agents, antioxidants, free radical disruptors, opacifiers, stabilizers, emollients, silicones, α-hydroxy acids, Antifoaming agents, moisturizing agents, vitamins, insect repellents, flavoring agents, preservatives, surfactants, anti-inflammatory agents, substance P antagonists, fillers, polymers, propellants, basicizing or acidifying agents, or coloring agents may be further included.

Hereinafter, a preferred embodiment of the present invention will be described in detail. However, the present invention is not limited to the embodiments described herein and may be embodied in other forms. In the embodiments introduced herein, the contents to be introduced are thorough and complete, and are provided to sufficiently convey the spirit of the present invention to those skilled in the art.

Example 1. Preparation of Shini extract

The dried Shini was purchased and the powder was prepared using a grinder. Shini powder 200g in 2kg of 30% ethanol, and stirred for 3 hours by heating with a heavy bath, Whatman No. 1 The extraction solution was filtered using filter paper. Thereafter, the supernatant was taken using a centrifuge, and the remaining residue was extracted twice by the same method. The filtered extract solution was freeze-dried to recover the dried powder of ethanol extract from Shin.

Example 2. SOCE inhibition of Sini extract through whole cell patch clamp

In order to confirm whether or not to inhibit SOCE (Store Operated Calcium Entry) of the Shini extract prepared in Example 1, a whole cell patch clamp using a HEK293T cell line overexpressed with ORAI1 and STIM1 proteins that are components of SOCE Was performed.

The depletion of calcium storage inside the ER occurs by IP ₃ in the cell. At this time, a protein called STIM1, which is the ER calcium sensor, is activated, and when ER calcium secretion occurs, a complex of ORAI1 and STIM1 is formed and the Orai-1 channel As activity occurs, the influx of calcium into cells through SOCE occurs. When I _SOCE generated by this mechanism was maintained in an equilibrium state, it was confirmed that the current of I _SOCE was suppressed by treating the extract, and also treated with 10 uM of BTP2, an ORAI1 inhibitor, to compare the relative activity of I _SOCE by each herbal extract. The degree of inhibition was comparatively analyzed.

Specifically, HEK293T cells (catalog no. CRL-3216) were used for pre-sale in the American type culture collection (ATCC, Manassas, VA, USA). The current measured was amplified using an Axopatch 200B (Molecular Device) amplifier, digitized through a Digidata 1440A (Molecular Device), and recorded with pClamp 10.4 (Molecular Device). To measure the current through SOCE, 135 mM CsOH, 125 mM Glutamic acid, 20 mM HEPES, 20 mM BAPTA, 3 mM MgATP, 1 mM MgCl ₂ , 0.002 mM Na-pyruvate (pH7.2 CsOH) is used as a pipette solution. As an extracellular solution, a solution of 135 mM NaCl, 3.6 mM KCl, 10 mM HEPES, 10 mM CaCl ₂ , 1 mM MgCl ₂ and 5 mM glucose (pH 7.4 NaOH) was used. In addition, to record each current, the voltage was fixed at -100 mV, and the inclined voltage was changed from -130 mV to -70 mV at intervals of 20 seconds for 100 ms. Inositole was added to the intracellular solution to activate SOCE. -3-phosphate (IP ₃ ) was added.

As a result, as shown in Figs. 1 to 3, 1 mg/ml of _Shini extract has I _SOCE It was confirmed that the activity was effectively suppressed.

Example 3. Confirmation of inhibition of TRPV1 activity of Sini extract through whole cell patch clamp

In order to confirm whether the Shini extract prepared in Example 1 can inhibit TRPV1, which is one of the calcium channels, a whole cell patch clamp was performed using a HEK293T cell line overexpressed with human TRPV1 protein.

Specifically, after confirming that the ion current (I _TRPV1 ) through _TRPV1 is increased by treating 1 uM of capsaicin capable of activating TRPV1, 10, 30, and 100 ug/ml of Shini extract were sequentially processed.

As a result, as can be seen in Figures 4 and 5, it was confirmed that the Shinyi extract inhibits the activity of I _TRPV1 in a concentration-dependent manner. That is, through the above results, it was confirmed that the Shinyi extract can more sensitively suppress the inward current of I _TRPV1 related to the influx of calcium into the cells.

Example 4. Confirmation of human T cell proliferation inhibitory effect

As confirmed in Examples 2 and 3, it was confirmed whether the extract of Shin, which can inhibit the influx of calcium into cells, can substantially inhibit the proliferation of human T cells.

Specifically, the blood mixed with 1:1 blood and phosphate buffer (PBS) in a 50ml tube containing 15ml Ficoll (GE Healthcare) was slowly added so as not to mix with Ficoll, and centrifuged at 300xg for 20 minutes before PBMC ( Peripheral Blood Mononuclear cell) layer was carefully collected and then centrifuged once more for 200xg and 10 min for platelet removal. After removing the platelets of the supernatant and resuspending the collected cell mass in PBS, T cells were separated from the separated PBMC using a Pan T cell isolation Kit (Milteny biotech). The separation process was performed according to the method provided by the manufacturer.

In order to confirm the proliferation of the isolated T cells, 2 μM of a fluorescence-indicating substance, carboxy-fluorescein succinimidyl ester (CFSE), was added to T cells and incubated at room temperature for 10 minutes. The labeled T cells were dispensed into a 96 well plate to be 2×10 ⁵ cells/well, and 3 μm/ml Anti-Human CD3 and 2 μg/ml Anti-Human CD28 were added to RPMI 1640. Cultured daily. After 3 days, only CD4 ⁺ T cells were stained using Anti-Human CD4-APC (BD Pharmingen), and then cell proliferation was analyzed using FACS.

As a result of experiments with 0.1, 0.3, and 1 mg/ml of Sini extract (MD), as shown in FIG. 6, it was confirmed that Sini extract inhibits the proliferation of human T cells in a concentration-dependent manner.

Example 5. Inhibition of ANO1 activity of Sini extract through whole cell patch clamp

The ANO1 ion channel, which is a calcium-dependent chlorine ion channel, is a very important ion channel for the secretion of mucus and electrolytes from the nasal epithelial cells, that is, the formation of a runny nose, so that the human ANO1 protein is used to determine how the new extract affects the human ANO1 ion channel. Whole cell patch clamp was performed using the overexpressed HEK293T cell line.

As a result, as can be seen in FIGS. 7 to 9, it was confirmed that the ion current through ANO1 was suppressed in a concentration-dependent manner when Shin treated with 0.1, 0.3, and 1 mg/ml extracts. It was confirmed that the inhibition was at least 87%.

Example 6. Calcium-dependent chlorine ion channel activity inhibition of Sini extract in human airway epithelial cells

Human airway epithelial cells have various chlorine ion channels including ANO1 ion channels, so IL-4, a cytokine secreted by Th2 T cells using human airway epithelial cell line Calu-3 (ATCC), is involved in runny nose production. It was confirmed whether the expression of the chlorine ion channel is increased and whether the extract of Shin can effectively suppress this.

As a result, as can be seen in FIG. 10, when the magnitude of the chlorine ion current through the calcium-dependent chlorine ion channel was measured 24 hours after treatment with IL-4 on Calu-3, it was confirmed that the magnitude of the current increased more than twice. Did.

On the other hand, when ANI-9, a selective inhibitor of the ANO1 ion channel, was treated, the current was completely suppressed, and it was confirmed that IL-4 selectively increases ANO1 expression.

In addition, when Shin treated with 0.1, 0.3 and 1 mg/ml of extract, it was confirmed in FIG. 11 that the expression of IL-4 could suppress the ionic current (I _CaCC ) through the calcium-dependent chlorine ion channel with increased expression. As shown, it was confirmed that I _CaCC is effectively suppressed due to the extract treatment.

Example 7. Confirmation of allergic rhinitis inhibitory effect of Shini extract

<7-1> Animal model and reagent preparation

A female 7-week-old Balb/c mouse was purchased (Orient Bio, Gyeonggi-do, Korea), reared under free-feeding conditions, stabilized in a breeding room for one week, and used in the experiment. Ovalbumin (OVA, sigma A5503), Inject Alum (Alum, Thermo scientific Prod#77161), Shini extract and hydrocortisone (Hydrocortisone, sigma H0888) were used as target substances to confirm the inhibitory effect of allergic rhinitis. Six mice were used.

<7-2> Cause allergic rhinitis in animal models

On Day 0, Day 7, and Day 14, 0.2 ml of the antigen was injected intraperitoneally once a week for 3 weeks. The normal control (NL) was injected with 200 μl of DPBS containing 2 mg of Alum, and the positive control (PC) and experimental group were injected with 25 μg of OVA and 200 μl of DPBS containing Alum 2 mg.

From Day 21, nasal drops were applied once a day for 8 days. 30 μl of DPBS as a normal control group and 30 μl of DPBS including 100 μg of OVA were added to the nasal cavity in the positive control group and the experimental group.

<7-3> Administration of therapeutic substances in animal models

From Day 24, nasal drops were applied 6 hours before the OVA challenge. DPBS contains 30% ethanol extracts of 0.001 mg/ml, 0.01 mg/ml, 0.1 mg/ml, 0.3 mg/ml, 0.8 mg/ml, 1 mg/ml, 3 mg/ml and 10 mg/ml, respectively. 20 μl of the solution was added to the nasal cavity once, and Hydrocortisone was added to the nasal cavity with 1 mg/ml included in 20 μl of DPBS.

<7-4> Evaluation method for allergic rhinitis in animal models

On day 28 (8 days of OVA challenge), after intranasal OVAchallenge without 30% ethanol extract or hydrocortisone treatment, one was placed in a clear acrylic box (15 cm x 20 cm x 15 cm) and the number of sneezing and scratching the nose was observed for 10 minutes. Each number was summed to show the result.

<7-5> How to measure cytokines in animal models

After separating the spleen cells of the mouse into single cells, lysing red blood cells, OVA 1 mg/ml was added to 5 x 10 ⁶ cells/ml of spleen cells, and cultured at 37.5° C. for 72 hours, followed by supernatant of the culture. The concentration of IL-4 was measured using Magnetic Luminex Assay (R&D Systems ^® , Minneapolis, MN, USA) according to the manufacturer's protocol (Cytokine measurements).

<7-6> Confirmation of the inhibitory effect of Xinyi extract on allergic rhinitis in animal models

As a result of evaluating allergic rhinitis in the animal model by the method of Example <7-4>, as can be seen in FIG. 12, 30% ethanol extract from Shin is 0.01 mg/ml, 0.1 mg/ml, 0.3 mg/ml , It was confirmed that the number of sneezing and scratching the nose was reduced by about 50% in mice dripping the group containing 0.8 mg/ml.

On the other hand, as a result of measuring the cytokines IL-4 and IL-13 in the animal model by the method of Example <7-5>, as can be seen in Figures 13 and 14, Shinyi extract 0.01 mg / ml, It was confirmed that the concentration of IL-4 in the mice dropping the group containing 0.1 mg/ml, 0.3 mg/ml, and 0.8 mg/ml was concentration-dependent. In addition, in IL-13, it was confirmed that the extract of Shini decreased production from 001 mg/ml to 08 mg/ml to a significant level compared to the positive control.

So far, the present invention has been focused on the preferred embodiments. Those skilled in the art to which the present invention pertains will understand that the present invention may be implemented in a modified form without departing from the essential characteristics of the present invention. Therefore, the disclosed embodiments should be considered in terms of explanation, not limitation. The scope of the present invention is shown in the claims rather than the foregoing description, and all differences within the equivalent range should be construed as being included in the present invention.

Claims (2)

1. Inhaled aerosol formulation for the prevention and treatment of allergic rhinitis, containing 0.001% to 0.08% by weight of the extract of Sini.
2. The aerosol formulation for inhalation according to claim 1, wherein the aerosol formulation for inhalation contains 0.01% by weight to 0.08% by weight of the Sini extract.

Images