KR102218823B1 - Composition for anti-allergy comprising extract of lithospermum erythrorhizon - Google Patents

Abstract

Disclosed herein are a composition for anti-allergy comprising a chichi extract as an active ingredient and a method for preparing an anti-allergy composition with suppressed cytotoxicity. The above extract does not contain siconine, and one or more siconine derivatives contained in chichi have been removed, or extraction of one or more siconine derivatives contained in chichi is suppressed. There are also no side effects.

Description

Anti-allergic composition containing chichi extract {COMPOSITION FOR ANTI-ALLERGY COMPRISING EXTRACT OF LITHOSPERMUM ERYTHRORHIZON}

Disclosed herein are a composition for anti-allergy comprising a chichi extract as an active ingredient and a method for preparing an anti-allergy composition with suppressed cytotoxicity.

Allergic rhinitis causes symptoms such as sneezing, clear runny nose, and stuffy nose, and at the same time, the head is heavy and tears come out. It often appears during the change of seasons or cold seasons of the year, and is often confused with a cold, which is different from a cold, and is often accompanied by asthma or hives.

Allergic rhinitis is one of the autoimmune diseases that accompany an inflammatory response.In patients with allergic rhinitis, interleukin-4 (IL-4), interleukin-5 (IL-5), and interleukin-13 (IL- 13) is known to increase.

Currently, anti-allergic drugs, antihistamines, antibiotics and anti-inflammatory drugs are prescribed for allergic rhinitis. Histamine secreted by cellular immune reactions induces inflammation of blood vessels. Under normal circumstances, histamine secretion helps prevent infection, but if it reacts to antigens such as pollen, it causes allergies. At this time, antihistamines act in a way that suppresses histamine secreted from cells to prevent infection.In general, antihistamines do not have a high therapeutic effect on allergic rhinitis, and there is drowsiness, dizziness, dry mouth, excitement, loss of appetite. There may be side effects such as abdominal pain, constipation, etc. And it is known to have no effect on nasal congestion. In addition, antihistamines are not fundamental treatments, but because they temporarily control symptoms, the efficacy is 12-24 hours. Therefore, there is a problem that antihistamines should be taken continuously. When taking antihistamines, they are metabolized to the liver or kidneys and absorbed into the body, so it is not desirable to eat them every day.

In addition, as one of the prescriptions for allergic rhinitis, topical steroids sprayed on the nose are used to eliminate nasal congestion, which also eliminates symptoms at the moment of use, but if used for a long time, it can cause sinusitis. Don't do it. Antibiotics and anti-inflammatory drugs also only serve to relieve temporary symptoms. As described above, since a method for curing allergic rhinitis is not yet known, there is a high demand for a therapeutic agent for allergic rhinitis that has no side effects and has excellent therapeutic effect.

KR 10-2011-0077131 A

In one aspect, the present specification aims to provide an anti-allergic composition that is safe for the human body because it is not cytotoxic and has no side effects even when taken daily.

In another aspect, the present specification aims to provide a method of preparing a composition for anti-allergy without cytotoxicity.

In another aspect, an object of the present specification is to provide a method of removing siconine and one or more siconine derivatives contained in chichi.

In one aspect, the technology disclosed in the present specification provides a composition for anti-allergy comprising an extract of Lithospermum erythrorhizon as an active ingredient.

In an exemplary embodiment, the extract may be a reflux extract of Chichi.

In an exemplary embodiment, the extract may be a reflux extract at 60° C. or higher.

In an exemplary embodiment, the extract may be a reflux extract using a lower alcohol solution having 1 to 4 carbon atoms as a solvent.

In an exemplary embodiment, the extract may be extracted after washing the chichi with a lower alcohol solution having 1 to 4 carbon atoms.

In an exemplary embodiment, the lower alcohol concentration of 1 to 4 carbon atoms in the solution may be 70 to 95% (v/v).

In an exemplary embodiment, the extract may be an extract of chichi root.

In an exemplary embodiment, the extract may have cytotoxicity suppressed.

In an exemplary embodiment, the extract may not contain siconine.

In an exemplary embodiment, the extract may be one from which one or more siconine derivatives contained in chichi have been removed or one or more siconine derivatives contained in chichi may be suppressed.

In an exemplary embodiment, the sikonin derivative is acetyl sikonin, isobutyrylshikonin, β,β-dimethylacryloyl sikonin (β,β-dimethylacryloylshikonin), isovaleryl siconin It may be one or more selected from the group consisting of (Isovalerylshikonin) and (α-methylbutyryl)shikonin ((α-methylbutyryl)shikonin).

In an exemplary embodiment, the extract is rosmarinic acid, N,N'-dicoumaroylspermidine (N,N'-dicoumaroylspermidine), lithospermic acid, salvianolic acid A One or more selected from the group consisting of (salvianolic acid A) and anhydroalkannin may be extracted.

In one exemplary embodiment, the composition is allergic rhinitis, allergic asthma (bronchial asthma), hay fever, Quincke's edema, anaphylaxis (anaphylaxis), hives, allergic Conjunctivitis (allergic conjunctivitis), allergic keratitis (allergic keratitis) and atopic dermatitis (atopic dermatitis) may be one or more preventive or therapeutic pharmaceutical composition selected from the group consisting of.

In one exemplary embodiment, the composition is allergic rhinitis, allergic asthma (bronchial asthma), hay fever, Quincke's edema, anaphylaxis (anaphylaxis), hives, allergic Conjunctivitis (allergic conjunctivitis), allergic keratitis (allergic keratitis) and atopic dermatitis (atopic dermatitis) may be one or more plant composition for improvement selected from the group consisting of.

In another aspect, the techniques disclosed herein include cleaning abutments; And reflux extraction of the washed chichi to obtain an extract, wherein the extract does not contain siconine, and provides a method of preparing an anti-allergy composition with suppressed cytotoxicity.

In an exemplary embodiment, the weary may be a root of weir.

In an exemplary embodiment, the extract may be an extract of a pulverized product obtained by pulverizing after washing the chichi.

In an exemplary embodiment, the washing step may be washing the ground teeth with a lower alcohol solution having 1 to 4 carbon atoms.

In an exemplary embodiment, the extraction step may be reflux extraction at 60° C. or higher.

In an exemplary embodiment, the extraction step may be reflux extraction using a lower alcohol solution having 1 to 4 carbon atoms as a solvent.

In an exemplary embodiment, the lower alcohol concentration of 1 to 4 carbon atoms in the solution may be 70 to 95% (v/v).

In one aspect, the technology disclosed in the present specification has the effect of providing an anti-allergic composition that is safe for the human body because it is not cytotoxic and has no side effects even when taken daily.

In another aspect, the technology disclosed in the present specification has the effect of providing a method for preparing the anti-allergy composition without cytotoxicity.

In another aspect, the technology disclosed herein has the effect of providing a method of removing sikonin and one or more siconin derivatives contained in chichi.

1 shows the results of HPLC analysis of Chichi extract according to Example 1 of the present specification.
2 shows the results of HPLC analysis of Chichi extract according to Comparative Example 1 of the present specification.
3 shows the results of HPLC analysis of Chichi extract according to Comparative Example 2 of the present specification.
Figure 4 shows the calibration curve of the siconine compound.
Figure 5 is in one experimental example of the present specification (1) 50 uM sikonin, (2) the extract of Comparative Example 1 at a concentration of 50 ug / ml, (3) the extract of Comparative Example 2 at a concentration of 50 ug / ml, ( 4) It shows a photograph comparing the extract of Example 1 at a concentration of 50 ug/ml with the naked eye.
6 is an example of (1) a control group, (2) 50 uM siconin, (3) an extract of Comparative Example 1 at a concentration of 50 ug/ml, and (4) an Example at a concentration of 50 ug/ml It shows the results of comparing the effect of each substance on apoptosis by treating the extract of 1 on macrophages (***; P<0.001, **; P<0.01).
Figure 7 shows the results of confirming the effect of the extract on allergic rhinitis according to an experimental example of the present specification.
Figure 8 shows the results of confirming the effect of the extract on allergic rhinitis disease according to an experimental example of the present specification (***; P<0.001, **; P<0.01, *; P<0.05).
9 shows the results of confirming the inhibitory effect of inflammation of Chichi extract in macrophages according to an experimental example of the present specification (***; P<0.001, **; P<0.01).
10 shows the results of confirming the allergy inhibitory effect of Chichi extract in mast cells according to an experimental example of the present specification (**; P<0.01, *; P<0.05).

Hereinafter, the present invention will be described in detail.

In one aspect, the technology disclosed in the present specification provides a composition for anti-allergy comprising an extract of Lithospermum erythrorhizon as an active ingredient.

Chichi ( Lithospermum erythrorhizon ) is a perennial herb that grows in the grasslands of mountains and fields in Korea, and there is no restriction on how to obtain the chichi, and it can be cultivated or used commercially.

In the present specification, "active ingredient" refers to an ingredient that exhibits a desired activity alone or can exhibit activity with a carrier that is not active as such.

In the present specification, "allergy" refers to a phenomenon in which an organism reacts in vivo due to an antigen-antibody reaction in contact with a certain foreign substance (allergen). There is no limitation on the type of the foreign substance, and specifically, may include pollen, drugs, vegetable fibers, fine dust, bacteria, food, dyes, or chemicals, but is not limited thereto.

In the present specification, the term "for the prevention or treatment of allergies" includes allergies and prevention of allergy-related diseases or uses thereof. The method of prevention or treatment is not limited, and for example, the disease can be prevented by fundamentally inhibiting the generation of allergens, but is not limited thereto.

In the present specification, "for allergy improvement" includes all use of reducing or alleviating the degree of allergy and allergy-related diseases. Specifically, it may include lowering or alleviating the degree of a disease that has already occurred, and considering that the composition of the present specification fundamentally inhibits allergens, it will be possible to prevent allergy and allergy-related diseases, but is limited thereto. It is not.

In the present specification, "extract" refers to a processed product of the extract, for example, drying, concentration, fractionation, fermentation, etc., to include all forms by further processing.

In the present specification, "reflux extract" refers to a substance obtained by extracting components contained in the extraction raw material by adding an extraction solvent and energy to the extraction raw material, and the vapor generated by the energy applied to the reaction vessel is cooled to a liquid and falls back into the reaction vessel. It means that it was manipulated and extracted to be able to. For example, the reflux extract can be obtained using a reflux extractor designed so that the vapor generated in the reaction vessel is cooled to become a liquid and flows back to the reaction vessel. The reflux temperature means the temperature at which the extraction solvent used during reflux extraction is boiled.

In an exemplary embodiment, the extract may be a reflux extract at 60° C. or higher, and more specifically 60 to 80° C. In another exemplary embodiment, the extract may be refluxed at a temperature of 60°C or higher, 65°C or higher, 70°C or higher, or 75°C or higher and 80°C or lower, 75°C or lower, 70°C or lower, or 65°C or lower. Preferably, the extract is a reflux extract at 70 to 75 ℃ may be preferred in implementing the effect of the present specification.

In an exemplary embodiment, the extract may be a reflux extract using a lower alcohol solution having 1 to 4 carbon atoms as a solvent.

In an exemplary embodiment, the extract may be extracted after washing the chichi with a lower alcohol solution having 1 to 4 carbon atoms.

In an exemplary embodiment, the extract may be extracted after grinding chichi. The "pulverization" means that the extraction raw material is shredded, crushed, or pulverized to be finely chopped, crushed, crushed, ground, or pulverized to form a powder, flake, or fibrous form.

In an exemplary embodiment, the extract may be an extract of chichi root.

In an exemplary embodiment, the extract is an extract of pulverized chichi root after washing with a lower alcohol solution having 1 to 4 carbon atoms to remove siconin and one or more siconin derivatives contained in chichi, and allergic rhinitis effect It may be desirable in improving the.

In an exemplary embodiment, the lower alcohol concentration of 1 to 4 carbon atoms in the solution may be 70 to 95% (v/v). In another exemplary embodiment, the lower alcohol concentration of the solution having 1 to 4 carbon atoms is 70% or more, 75% or more, 80% or more, 85% or more, or 90% or more, and 95% or less, 90% or less, 85% It may be less than or equal to 80% or less than or equal to 75%. Preferably, the lower alcohol concentration of 1 to 4 carbon atoms in the solution may be 90 to 95% (v/v).

In an exemplary embodiment, the extract may have cytotoxicity suppressed.

In an exemplary embodiment, the extract does not contain siconine, and the extract may be one from which one or more siconine derivatives contained in chichi have been removed, or that the extraction of one or more siconine derivatives contained in chichi is suppressed. have.

Accordingly, the Chichi extract according to the present specification, unlike the conventional Chichi extract, has an effect of maximizing anti-allergic efficacy, especially allergic rhinitis, while suppressing cytotoxicity and without side effects even if taken for a long time.

In an exemplary embodiment, the sikonin derivative is acetyl sikonin, isobutyrylshikonin, β,β-dimethylacryloyl sikonin (β,β-dimethylacryloylshikonin), isovaleryl siconin It may be one or more selected from the group consisting of (Isovalerylshikonin) and (α-methylbutyryl)shikonin ((α-methylbutyryl)shikonin).

In an exemplary embodiment, the extract is acetylshikonin, isobutyrylshikonin, β,β-dimethylacryloylshikonin (β,β-dimethylacryloylshikonin), isovalerylshikonin (Isovalerylshikonin) ) And (α-methylbutyryl)shikonin ((α-methylbutyryl)shikonin) may not be included.

In an exemplary embodiment, the extract is rosmarinic acid, N,N'-dicoumaroylspermidine (N,N'-dicoumaroylspermidine), lithospermic acid, salvianolic acid A One or more selected from the group consisting of (salvianolic acid A) and anhydroalkannin may be extracted.

Accordingly, the Chichi extract according to the present specification, unlike the conventional Chichi extract, has an effect of maximizing anti-allergic efficacy, especially allergic rhinitis, while suppressing cytotoxicity and without side effects even if taken for a long time.

In one exemplary embodiment, the composition is allergic rhinitis, allergic asthma (bronchial asthma), hay fever, Quincke's edema, anaphylaxis (anaphylaxis), hives, allergic Conjunctivitis (allergic conjunctivitis), allergic keratitis (allergic keratitis) and atopic dermatitis (atopic dermatitis) may be one or more preventive or therapeutic pharmaceutical composition selected from the group consisting of.

In one exemplary embodiment, the composition is allergic rhinitis, allergic asthma (bronchial asthma), hay fever, Quincke's edema, anaphylaxis (anaphylaxis), hives, allergic Conjunctivitis (allergic conjunctivitis), allergic keratitis (allergic keratitis) and atopic dermatitis (atopic dermatitis) may be one or more plant composition for improvement selected from the group consisting of.

In one exemplary embodiment, the formulation of the pharmaceutical composition may be a solution, suspension, emulsion, gel, drop, cream, ointment, patch, pad, or spray, but is not limited thereto. The formulation can be easily prepared according to conventional methods in the art, and excipients, hydrating agents, emulsification accelerators, suspending agents, salts or buffers for controlling osmotic pressure, coloring agents, spices, stabilizers, preservatives, preservatives or other compatible Supplements can be used in moderation.

In addition, the pharmaceutical composition may be administered orally, parenteral, rectal, topical, transdermal, intravenous, intramuscular, intraperitoneal, subcutaneous, etc. according to a desired method, and the active ingredient of the pharmaceutical composition is the age of the subject to be administered. , Sex, weight, pathological condition and severity, route of administration, or the judgment of the prescriber. Determination of the application amount based on these factors is within the level of those skilled in the art, and its daily dosage is, for example, 0.001 mg/g/day to 100 mg/g/day, more specifically 0.5 mg/g/day to 50 mg/day. It may be g/day, but is not limited thereto.

The food composition herein may provide various types of food additives or functional foods. Specifically, it may be processed into leached tea, liquid tea, beverage, fermented milk, cheese, yogurt, juice, probiotics or health supplements, etc. including the composition, and may be used in the form of various other food additives.

In addition, the food composition may further include other ingredients that can give a synergistic effect to the main effect within a range in which the active ingredient does not impair the intended main effect. For example, to improve physical properties, additives such as fragrances, coloring agents, disinfectants, antioxidants, preservatives, moisturizers, thickeners, inorganic salts, emulsifiers, or synthetic polymer materials may be further included. In addition, auxiliary ingredients such as water-soluble vitamins, oil-soluble vitamins, polymer peptides, polymer polysaccharides or seaweed extract may be further included. The above ingredients may be appropriately selected and blended by a person skilled in the art according to the formulation or purpose of use, and the amount added may be selected within a range not impairing the purpose and effect of the present specification. For example, the amount of the components added may range from 0.001 to 5% by weight, or from 0.01 to 3% by weight, based on the total weight of the composition.

In an exemplary embodiment, the extract may be included in an amount of 0.001 to 80% by weight based on the total weight of the composition. More specifically, the extract is 0.001 wt% or more, 0.01 wt% or more, 0.1 wt% or more, 1 wt% or more, 2 wt% or more, 3 wt% or more, 4 wt% or more, 5 wt% based on the total weight of the composition % Or more, 6% or more, 7% or more, 8% or more, 9% or more, or 10% or more and 80% or less, 70% or less, 60% or less, 50% or less, 40% by weight % Or less, 30 wt% or less, 20 wt% or less, 10 wt% or less, or 5 wt% or less.

In another aspect, the techniques disclosed herein include cleaning abutments; And reflux extraction of the washed chichi to obtain an extract, wherein the extract does not contain siconine, and provides a method of preparing an anti-allergy composition with suppressed cytotoxicity.

In another aspect, the techniques disclosed herein include cleaning the abutment; And it provides a method for removing the siconine and one or more siconine derivatives contained in the chichi, comprising the step of obtaining an extract by reflux extraction of the washed chichi.

In an exemplary embodiment, the sikonin derivative is acetyl sikonin, isobutyrylshikonin, β,β-dimethylacryloyl sikonin (β,β-dimethylacryloylshikonin), isovaleryl siconin It may be one or more selected from the group consisting of (Isovalerylshikonin) and (α-methylbutyryl)shikonin ((α-methylbutyryl)shikonin).

In an exemplary embodiment, the weary may be a root of weir.

In an exemplary embodiment, the extract may be an extract of a pulverized product obtained by pulverizing after washing the chichi.

In an exemplary embodiment, the washing step may include washing the branch teeth with a lower alcohol solution having 1 to 4 carbon atoms. The thus prepared Chichi extract, unlike conventional Chichi extract, has an effect of maximizing anti-allergic efficacy, especially allergic rhinitis, while suppressing cytotoxicity and having no side effects even when taken for a long time.

In an exemplary embodiment, in the washing step, washing the branch by adding a lower alcohol solution having 1 to 4 carbon atoms corresponding to the weight of branch, specifically 8 to 12 times the dry weight of the branch root. It may be desirable in removing sikonin and one or more siconin derivatives and implementing the efficacy of improving allergic rhinitis.

In an exemplary embodiment, the washing step is performed at room temperature, such as, but not limited to, 15 to 25° C. in removing sikonin and one or more siconin derivatives contained in chichi and improving allergic rhinitis It may be desirable.

In an exemplary embodiment, in the washing step, washing the chichi for 60 seconds to 300 seconds may be preferable in removing siconine and one or more siconine derivatives contained in the chichi and implementing the efficacy of improving allergic rhinitis. In another exemplary embodiment, the washing time is 60 seconds or more, 70 seconds or more, 80 seconds or more, 90 seconds or more, 100 seconds or more, 110 seconds or more, 120 seconds or more, 130 seconds or more, 140 seconds or more, or 150 seconds. It may be more than 300 seconds, 290 seconds or less, 280 seconds or less, 270 seconds or less, 260 seconds or less, 250 seconds or less, 240 seconds or less, 230 seconds or less, 220 seconds or less, 210 seconds or less, or 200 seconds or less. . If the cleaning time is out of the above range, the specific component to be removed may not be removed or a specific component that is not intended may be removed, and thus, fatigue for 60 seconds to 300 seconds, more specifically 60 seconds to 240 seconds, or 60 seconds to 120 seconds It may be desirable to wash.

In an exemplary embodiment, the extraction step may be reflux extraction at 60° C., more specifically 60 to 80° C. In another exemplary embodiment, the extraction step may be reflux extraction at a temperature of 60°C or higher, 65°C or higher, 70°C or higher, or 75°C or higher and 80°C or lower, 75°C or lower, 70°C or lower, or 65°C or lower. have.

In an exemplary embodiment, the extraction step is performed for 200 minutes or more, preferably 240 minutes to 300 minutes, in removing sikonin and one or more siconin derivatives contained in chichi and implementing the efficacy of improving allergic rhinitis. It may be desirable.

In an exemplary embodiment, the extraction step may be reflux extraction using a lower alcohol solution having 1 to 4 carbon atoms as a solvent.

In an exemplary embodiment, the lower alcohol having 1 to 4 carbon atoms may be ethanol.

In an exemplary embodiment, the lower alcohol concentration of 1 to 4 carbon atoms in the solution may be 70 to 95% (v/v). In another exemplary embodiment, the lower alcohol concentration of the solution having 1 to 4 carbon atoms is 70% or more, 75% or more, 80% or more, 85% or more, or 90% or more, and 95% or less, 90% or less, 85% It may be less than or equal to 80% or less than or equal to 75%. Preferably, the lower alcohol concentration of 1 to 4 carbon atoms in the solution may be 90 to 95% (v/v).

Hereinafter, the present invention will be described in more detail through examples. These examples are for illustrative purposes only, and it will be apparent to those of ordinary skill in the art that the scope of the present invention is not construed as being limited by these examples.

Example 1. Preparation of reflux extract after washing of chichi root

Chichi extract was prepared as follows using chichi purchased in Jeongseon, Gangwon-do in August 2016. First, 20 kg of dried chichi roots were washed three times at room temperature for 2 minutes using 300 ml of a 95% (v/v) ethanol solution per 30 g. Thereafter, the chichi roots were pulverized finely, and a 95% (v/v) ethanol solution was added to the crushed chichi roots, followed by extraction three times by reflux extraction at 70° C. for 4 hours. The ethanol solution was all removed under vacuum from the Jichi extract obtained through a series of processes as described above to obtain 1393.5 g of Jichi extract.

Comparative example One. Exhaustion Root Cold Extract manufacturing

Chichi extract was prepared as follows using chichi purchased in Jeongseon, Gangwon-do in August 2016. 1 kg of dried chichi roots were pulverized finely, and 95% (v/v) ethanol solution was added to the crushed chichi roots, followed by stirring at room temperature for 24 hours to extract by cold extraction method (cold extraction method). The ethanol solution was all removed under vacuum from the Jichi extract obtained through a series of processes as described above to obtain 26 g of Jichi extract.

Comparative Example 2. Preparation of reflux extract of Chichi root

Chichi extract was prepared as follows using chichi purchased in Jeongseon, Gangwon-do in August 2016. 33 g of dried chichi roots were pulverized finely, and a 95% (v/v) ethanol solution was added to the crushed chichi roots, followed by extraction with reflux at 70° C. for 4 hours (reflux extraction method) three times. The ethanol solution was removed under vacuum from the Jichi extract obtained through a series of processes as described above to obtain 3 g of Jichi extract.

Experimental Example 1. Comparative Analysis of Components of Chichi Extract

Each component was compared and analyzed using HPLC, LC-MS, and NMR analysis for the extracts prepared in Example 1 and Comparative Examples 1 and 2. The extracts of Example 1, Comparative Example 1, and Comparative Example 2 were loaded at 10 mg/ml, 10 mg/ml, and 20 mg/ml, respectively, and HPLC analysis was performed. Table 1 below shows the HPLC analysis conditions, and Table 2 shows the compound names corresponding to each peak number in FIGS. 1 to 3.

Time Water
(0.1% Formic acid) Acetonitrile
(0.1% Formic acid) 0 95 5 2 95 5 40 50 50 60 50 50 61 30 70 71 30 70 72 20 80 85 20 80 90 5 95 95 95 5 Column: Hydrosphere C18 (Ø4.6x250 mm, 5 μm)
Flow: 0.7 ml/min
Injection volume: 10 μL
Detection: 280 nm
Running time: 95 min
Column temperature: 25 °C

No. Compound name One N,N'-dicoumaroylspermidine 2 Rosmarinic acid 3 Lithospermic acid 4 Salvianolic acid A 5 Shikonin 6 Shikonofuran D 7 Shikonofuran E 8 β-hydroxyisovalerylshikonin 9 Acetylshikonin 10 Deoxyshikonin 11 Anhydroalkannin 12 Isobutyrylshikonin 13 β,β-dimethylacryloylshikonin 14 Isovalerylshikonin 15 (α-methylbutyryl)shikonin

1 to 3, since the maximum units of the Y-axis are 60 mAU, 1400 mAU, and 120 mAU, respectively, the amount of the material corresponding to the peak can be relatively compared based on this. In the extract of Comparative Example 1, sikonin and a number of siconin derivative components were detected and the content thereof was also high, whereas in the extract of Example 1, sikonin components were not detected, and the content as well as the type of siconine derivative components were significantly reduced. Confirmed. In the extract of Example 1, other components were detected instead of the siconine derivative component. On the other hand, it was confirmed that the extract of Comparative Example 2 obtained by reflux extraction without washing the chichi roots with an ethanol solution contains a siconine component and a higher content of the siconine derivative component, unlike the extract of Example 1.

In addition, the content of sikonin contained in each extract was calculated using the sikonin standard curve of FIG. 4 and shown in Table 3 below. The extracts of Example 1, Comparative Example 1, and Comparative Example 2 contained 0 ug, 21.5 ug, and 1.9 ug of siconine, respectively, per 1 mg of extract, and the extract of Example 1 subjected to the washing process contained siconine. It was confirmed that there was no.

Sikonin content in extract (%) Extract of Example 1 0.00 Extract of Comparative Example 1 2.15 Extract of Comparative Example 2 0.19

In addition, when the extract was visually checked, it was found that the types and contents of the siconine and siconine derivative components contained in each extract were different, so that all of them had different colors. It was confirmed that the extract of Example 1 had a color close to transparency because the siconine component was removed (see FIG. 5).

Experimental Example 2. Analysis of apoptosis of Chichi extract

Sikonin at a concentration of 50 uM, the extract of Comparative Example 1 at a concentration of 50 ug/ml, and the extract of Example 1 at a concentration of 50 ug/ml were treated with equal amounts of THP1 cells (human monocytes, macrophages) and cultured for 24 hours. After that, the degree of cell death was confirmed through MTT assay.

As a result, only about 60% of the cells were alive in the group treated with the extract of Comparative Example 1, but more than 95% of the cells were alive in the group treated with the extract of Example 1, so the extract of Example 1 had a very high cytotoxic inhibitory effect. It was confirmed to be excellent. In addition, even in the group treated with only sikonin, it was confirmed that only about 50% of the cells were alive, so that siconin exhibited cytotoxicity (see FIG. 6).

Experimental example 3. Exhaustion Extract of Anti-allergy Efficacy analysis

(1) Confirmation of the effect on allergic rhinitis disease using a mouse model

6-week-old female Balb/c mice weighing 20 g were purchased from Orient Bio and used, and were acclimated in a laboratory environment (temperature; 25±1° C., humidity; 40±5%) for 7 days after purchase.

Normal group (Control, PBS), allergic rhinitis group (OVA), ciclesonide (Ciclesonide) treatment group (OVA + CIC), the chichi extract treatment group of Example 1 (OVA + EELE) mice into four groups Classified and tested. Cyclesonide (Omnaris, Nycomed Canada Inc., Oakville, ON, Canada) is a glucocorticoid used to treat asthma and allergic rhinitis, and was used as a positive control. Allergic rhinitis was induced by sensitizing with OVA and Al(OH) ₃ and then spraying OVA into the nasal cavity.

Four groups of mice were treated with 100 ug of ovalbumin (OVA; grade V; Sigma-Aldrich, St. Louis, MO, USA) and 2 mg of aluminum hydroxide (Sigma Aldrich) in the abdominal cavity on days 0, 7 and 14. Injected. To all sensitized mice, 500 ug of OVA was administered intranasally every day for 7 days from the 21st day, and PBS was administered intranasally instead of OVA in the case of the normal group. In addition, in the case of the ciclesonide treatment group, 15 ug of ciclesonide was administered intranasally every day along with OVA administration. In addition, in the case of the chichi extract-treated group of Example 1, chichi extract was dissolved in 0.5% (v/v) CMC (carboxymethylcellulose) solution with OVA administration and orally administered at 50 mg/kg for 7 days. Mice on day 28 were sacrificed.

The number of eosinophils was quantified through H&E staining, and the number of eosinophils in the lamina propria was counted. Eosinophils were morphologically defined by the presence of bilobular nuclei and eosinophilic granules in the cytoplasm. As shown in the H&E staining result of FIG. 7, unlike the normal group, in the group in which allergic rhinitis was induced by OVA treatment, the number of eosinophils in the nasal mucosa intrinsic layer infiltrated significantly increased. In the Chichi extract-treated group of Example 1, it was confirmed that the infiltration of eosinophils was significantly suppressed.

Nasal mucosal goblet cells responsible for mucus and secretion were observed using the PAS technique. As a result of observing the germ cells stained with PAS staining along the nasal septal mucosa with an optical microscope, it was confirmed that the number of germ cells was remarkably increased in the allergic rhinitis-induced group. However, the number of germ cells did not increase in the cyclsonide treatment group and the Chichi extract treatment group of Example 1 (see Figs. 7 and 8).

In addition, mast cells can be identified using toluidin blue staining, and the number of mast cells was counted in the nasal septal mucosa with an optical microscope. Infiltration of mast cells also significantly increased when allergic rhinitis was induced, but it was confirmed that the infiltration of mast cells was significantly suppressed in the cyclsonide treatment group and the Chichi extract treatment group of Example 1 (see FIGS. 7 and 8 ).

As a result of observing the concentration of IL-5 in the serum through the ELISA technique, it was confirmed that the concentration of IL-5 was significantly increased in the allergic rhinitis-inducing group. However, in the cyclsonide treatment group and the Chichi extract treatment group of Example 1, the concentration of IL-5 was significantly lower than in the allergic rhinitis induction group (see FIG. 8).

As described above, it was confirmed that the extract of Example 1 had a siconin component removed and a sikonin derivative component removed or reduced, but it was confirmed that the eosinophilic inflammatory response was reduced and the allergic rhinitis inhibitory effect was very excellent.

(2) Confirmation of the anti-inflammatory effect of Chichi extract in macrophages through in vitro experiments

Reporter cell line THP1-Xblue (Invivogen, THP1-XBlue™ Cells, thpx-sp), THP1-Blue-, a reporter cell line that can confirm the degree of activation of NFkB/AP1 and IRF, transcription factors related to inflammation in human monocyte cells (THP1). Using ISG (Invivogen, THP1-Blue™ ISG Cells, thp-isg), it was confirmed whether the Chichi extract of Example 1 had an effect on inhibiting inflammation. The THP1-Xblue cell line and the THP1-Blue-ISG cell line were treated with 1 ug/ml of inflammatory substances LPS (Invivogen, Lipopolysaccharide, LPS from E. coli K12) and 1 ug/ml of Pam3CSK4 (Invivogen), respectively, and a reporter after 24 hours. The activity was measured as SEAP activity.

As a result, as shown in Figure 9, in the case of the group treated with LPS and Pam3, respectively, it was confirmed that the SEAP response was increased in both the THP1-Xblue cell line and the THP1-Blue-ISG cell line. However, in addition to this, when the Chichi extract of Example 1 was treated at a concentration of 10 ug/ml, 50 ug/ml, and 100 ug/ml, it was confirmed that the SEAP activity significantly decreased as the concentration increased. It was confirmed that the Chichi extract of Example 1, which was not contained, was effective in inhibiting inflammation.

(3) Confirmation of allergy inhibitory effect of Chichi extract in mast cells through in vitro experiments

Using the mast cell line RBL-2H3 cell line, it was tested whether the Chichi extract of Example 1 could inhibit the expression of IL-3, IL-4, and IL-13 known as factors related to allergic reactions. RBL-2H3 cells were activated through PMA (50 μg/ml)/ionomycin (0.1 μM) stimulation, and 6 hours later, the expression levels of IL-3, IL-4, and IL-13 compared to Gapdh were measured through quantitative realtime RT-PCR. Confirmed.

As a result, as shown in FIG. 10, the expression of IL-3, IL-4, and IL-13 increased by PMA/ionomycin was 50 μg/ml and 100 μg/ml of Chichi extract of Example 1 was added. When treated with, it was confirmed that the expression was suppressed as the concentration increased. Through this, it was confirmed that the Chichi extract of Example 1, which does not contain a sikonin component, has an allergy inhibitory effect in mast cells.

Examples of the formulation of the composition according to an aspect of the present specification are described below, but can be applied to various other formulations, and this is not intended to limit the present invention, but is intended to be described in detail.

[Formulation Example 1] Soft Capsule

150 mg of chichi extract of Example 1, 2 mg of palm oil, 8 mg of hard palm oil, 4 mg of sulfur wax, and 6 mg of lecithin were mixed, and 400 mg per capsule was filled according to a conventional method to prepare a soft capsule.

[Formulation Example 2] Tablet

150 mg of chichi extract, 100 mg of glucose, 50 mg of red ginseng extract, 96 mg of starch, and 4 mg of magnesium stearate of Example 1 were mixed, and 40 mg of 30% ethanol was added to form granules, followed by drying at 60°C and a tablet press. It was compressed into tablets using.

[Formulation Example 3] Granules

150 mg of chichi extract of Example 1, 100 mg of glucose, 50 mg of red ginseng extract, and 600 mg of starch were mixed to form granules, and then 100 mg of 30% ethanol was added to form granules, followed by drying at 60°C to form granules. Filled. The final weight of the contents was 1 g.

[Formulation Example 4] Drinks

150 mg of chichi extract of Example 1, 10 g of glucose, 50 mg of red ginseng extract, 2 g of citric acid, and 187.8 g of purified water were mixed and filled into a bottle. The final volume of the contents was 200 ml.

[Formulation Example 5] Preparation of health food

Chichi extract of Example 1............1000 mg

Vitamin mixture

Vitamin A acetate.....................70 ㎍

Vitamin E ....................................... 1.0 mg

Vitamin B1............0.13 mg

Vitamin B2 ..................................... 0.15 mg

Vitamin B6............................. 0.5 mg

Vitamin B12...................................... 0.2 ㎍

Vitamin C......................................... 10 mg

Biotin........................................... 10 ㎍

Nicotinic acid amide..................................1.7 mg

Folic acid............................................. 50 ㎍

Calcium pantothenate ...................................0.5 mg

Mineral mixture

Ferrous sulfate....................................... 1.75 mg

Zinc oxide........................................0.82 mg

Magnesium carbonate.....................................25.3 mg

Potassium Phosphate.....................................15 mg

Dicalcium Phosphate.....................................55 ㎎

Potassium citrate......................................90 mg

Calcium carbonate........................................100 mg

Magnesium chloride....................................24.8 mg

The composition ratio of the vitamin and mineral mixture is relatively suitable for health food, but it may be arbitrarily modified, and the above ingredients are mixed according to a conventional health food manufacturing method. Granules can be prepared and used for preparing health food compositions according to a conventional method.

[ Formulation example 6] Manufacture of health drinks

Chichi extract of Example 1 ........................1000 mg

Citric acid..............................1000 mg

Oligosaccharide............100 g

Plum concentrate......................................2 g

Taurine............ 1 g

Total by adding purified water..........900 ml

After mixing the above ingredients according to the normal health drink manufacturing method, stirring and heating at 85°C for about 1 hour, the resulting solution is filtered and obtained in a sterilized 2 liter container, sealed and sterilized, and then stored in a refrigerator. It was used to prepare a health drink composition according to the method.

The composition ratio is composed of ingredients suitable for a relatively preferred beverage in a preferred embodiment, but the mixing ratio may be arbitrarily modified according to regional and ethnic preferences, such as the demand class, the country of demand, and the intended use. Those of ordinary skill in the art to which the present invention belongs will be able to perform various applications and modifications within the scope of the present invention based on the above contents.

As described above, specific parts of the present invention have been described in detail, and for those of ordinary skill in the art, these specific techniques are only preferred embodiments, and the scope of the present invention is not limited thereby It will be obvious. Therefore, it will be said that the practical scope of the present invention is defined by the appended claims and their equivalents.

Claims (21)

Chichi ( Lithospermum erythrorhizon ) containing extract as an active ingredient,
The extract is a reflux extract extracted using an ethanol solution as a solvent after washing the chichi with an ethanol solution,
The extract is characterized in that it does not contain sikonin and sikonin derivatives,
The sikonine derivatives include acetylshikonin, isobutyrylshikonin, β,β-dimethylacryloylsikonin, isovalerylshikonin, and isovalerylshikonin. Methylbutyryl) sikonin ((α-methylbutyryl)shikonin) is one or more selected from the group consisting of, anti-allergic composition with suppressed cytotoxicity of chichi.
The method of claim 1,
The sikonine derivatives include acetylshikonin, isobutyrylshikonin, β,β-dimethylacryloylsikonin, isovalerylshikonin, and isovalerylshikonin. Methylbutyryl)shikonin ((α-methylbutyryl)shikonin), anti-allergic composition with suppressed cytotoxicity of chichi.
The method of claim 1,
The extract is a reflux extract at 60 ℃ or more, anti-allergic composition suppressed the cytotoxicity of Chichi.
delete delete The method of claim 1,
The concentration of the ethanol solution is 70 to 95% (v/v), anti-allergic composition in which the cytotoxicity of chichi is suppressed.
The method of claim 1,
The extract is an extract of chichi root, anti-allergic composition with suppressed cytotoxicity of chichi.
delete delete delete delete The method of claim 1,
The extracts include rosmarinic acid, N,N'-dicumaroylspermidine (N,N'-dicoumaroylspermidine), lithospermic acid, salvianolic acid A, and Hydroalkanin (anhydroalkannin) containing, anti-allergic composition suppressed the cytotoxicity of chichi.
The method of claim 1,
The composition is a pharmaceutical composition for the prevention or treatment of allergic rhinitis, anti-allergic composition with suppressed cytotoxicity of chichi.
The method of claim 1,
The composition is a food composition for improving allergic rhinitis, anti-allergic composition with suppressed cytotoxicity of chichi.
A method for preparing an anti-allergy composition in which the cytotoxicity of chichi according to any one of claims 1 to 3, 6, 7, and 12 to 14 is suppressed,
Washing the ground with an ethanol solution; And
Comprising the step of obtaining an extract by reflux extraction of the washed chichi using an ethanol solution as a solvent,
The extract is characterized in that it does not contain sikonin and sikonin derivatives,
The sikonine derivatives include acetylshikonin, isobutyrylshikonin, β,β-dimethylacryloylsikonin, isovalerylshikonin, and isovalerylshikonin. Methylbutyryl) sikonin ((α-methylbutyryl)shikonin) is one or more selected from the group consisting of, a method for producing a composition for anti-allergy suppressed cytotoxicity of chichi.
The method of claim 15,
The washing step is to wash for 60 seconds to 300 seconds at room temperature, the method for producing an anti-allergy composition in which the cytotoxicity of chichi is suppressed.
The method of claim 15,
The extract is an extract of the pulverized pulverized after washing the chichi, the method for producing an anti-allergy composition in which the cytotoxicity of chichi is suppressed. delete delete delete delete

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