KR20190136773A - Composition for alleviating histamine-independent itching comprising myrrh extract as effective component - Google Patents

Abstract

The present invention relates to a histamine-independent itch relief composition containing a myrrh extract as an active ingredient. The myrrh extract of the present invention may be effectively used in the development of cosmetics or therapeutics for the inhibition or alleviation of itching.

Description

Composition for alleviating histamine-independent itching comprising myrrh extract as effective component

The present invention relates to a histamine-independent itching relief composition containing a myrrh extract as an active ingredient.

Itching is a common sensation in skin diseases, which can be considered an unpleasant sensation that causes the urge to scratch or rub the skin. Itching varies greatly from person to person, and even if the same stimulus is given to the same person, the degree of itching can sometimes be different. Skin diseases in which itching is a major problem include atopic dermatitis, contact dermatitis, molar eczema, neurodermatitis, urticaria, dry skin, seborrheic dermatitis, and psoriasis, and the degree varies depending on the affected area and the patient's sensitivity. Can be.

Therapeutic agents known to be effective for itching include corticosteroids, antihistamines, immunosuppressants, and capsaicin. Steroids have an excellent effect on anti-inflammatory and immune suppression, but when used for a long time, the elasticity of the skin disappears, the skin becomes thinner, and the blood vessels are exposed, and in severe cases, hormones may cause systemic symptom, adrenal function and immunity. Degradation appears, the disadvantage of stopping the drug has the disadvantage of explosive deterioration. Antihistamines suppress the release of histamine from mast cells, thereby reducing itching, but when used for a long time, side effects such as anxiety and anorexia appear. Cyclosporine, an immunosuppressive agent, can cause serious side effects such as hypertension, nephrotoxicity, and drug interactions during systemic administration. Capsaicin is widely known to be the fastest and most potent anti-itch, but has limited use due to strong skin irritation.

These existing itch treatments have different effects depending on the individual's constitution, and in particular, since artificial chemical compositions are added during the manufacturing process and the manufacturing process, it is difficult to trust the safety. There is a need for further treatment.

Myrrh is a natural rubber resin obtained by wounding bark such as Commiphora myrrha or C. abyssinica , a native plant of the Bursaceae family in Africa and Arabia. Also called myrrha. Myrrh is known to be effective in releasing clumps of oriental medicine to promote blood circulation, reduce swelling, relieve pain, and give birth to new flesh.

Meanwhile, Korean Patent No. 1782962 discloses a composition for treating leukemia and a method for producing myrrh extracts, and Korean Patent No. 1508295 discloses a composition for improving itching, which contains a golden and gold silver extract. However, there is no description of the histamine-independent itching relief composition containing the myrrh extract of the present invention as an active ingredient.

The present invention is derived from the above requirements, the inventors of the present invention, myrrh extract is not cytotoxic, and excellent in inhibiting the production of IL (interleukin) -31 and extracellular signal-regulated kinase (ERK) phosphorylation and histamine- The present invention was completed by confirming to alleviate histamine-independent itching.

In order to solve the above problems, the present invention provides a cosmetic composition for suppressing or alleviating itching containing myrrh extract as an active ingredient.

The present invention also provides a pharmaceutical composition for suppressing or alleviating itching, containing a myrrh extract as an active ingredient.

Myrrh extract of the present invention has an excellent effect of inhibiting the expression of itching-related cytokines and inflammation-related proteins without toxicity or side effects, it may be useful as an active ingredient of the composition for suppressing or alleviating itching.

1 is a result of confirming the cytotoxicity of myrrh ethanol extract (ME).
Figure 2 is the result of analyzing the histamine production amount according to the myrrh ethanol extract (ME) treatment. PMA + A23187: Source of Inflammatory Response.
3 is a result of analyzing the inhibitory effect of IL-31 production of myrrh ethanol extract (ME) by real time-PCR (A) and Western blot (B).
Figure 4 is the result of analyzing the inhibitory effect of ERK, JNK, p38 (A) and NF-kB (B) of myrrh ethanol extract (ME).
5 is a result of analyzing the itching inhibitory effect of myrrh extract in a mouse animal model. control: untreated, Compound 48/80: itch inducer, prednisolone: itch suppression standard.

In order to achieve the object of the present invention, the present invention provides a cosmetic composition for suppressing or alleviating itching containing myrrh (myrrh) extract as an active ingredient.

As used herein, the term 'myrrh' refers to a natural rubber resin obtained by wounding bark such as Commiphora myrrha or C. abyssinica , which is a plant of the Burseraceae. it means.

In the cosmetic composition for suppressing or alleviating the itch of the present invention, the myrrh extract may be an extract of myrrh using water, a lower alcohol having 1 to 4 carbon atoms or a mixed solvent thereof, preferably extracted using ethanol May be, but is not limited thereto.

In the cosmetic composition according to an embodiment of the present invention, the myrrh extract is mixed with 90 ~ 110 g of myrrh and 80 ~ 99% (v / v) ethanol 1.9 ~ 2.1 L and left at room temperature for 2 to 4 days 0.45 ㎛ The filter may be prepared by filtration, and more specifically, 100 g of myrrh and 2 L of 90% (v / v) ethanol may be mixed and left for 3 days at room temperature, followed by filtration with a 0.45 μm filter. It is not limited.

In the pharmaceutical composition for suppressing or alleviating the itch of the present invention, the itch may be histamine-independent itching. The histamine-independent itch refers to an itch that is not mediated by histamine.

Cosmetic compositions for suppressing or alleviating itch of the present invention are selected from solutions, suspensions, emulsions, pastes, gels, creams, lotions, powders, soaps, surfactant-containing cleansing, oils, powder foundations, foundations, wax foundations and sprays It is preferably one of the formulations, and more preferably, external skin ointments, creams, supple cosmetics, nourishing cosmetics, packs, essences, hair tonics, shampoos, rinses, hair conditioners, hair treatments, gels, skin lotions, skin softeners , Skin toner, astringent, lotion, milk lotion, moisture lotion, nutrition lotion, massage cream, nutrition cream, eye cream, moisture cream, hand cream, foundation, nutrition essence, sunscreen, soap, cleansing foam, cleansing lotion, cleansing cream It may have any one formulation selected from, body lotion and body cleanser, but is not limited thereto. The composition of each of these formulations may contain various bases and additives necessary for the formulation of the formulation and are suitable, and the types and amounts of these components can be easily selected by those skilled in the art.

When the formulation of the present invention is a paste, cream or gel, animal carriers, plant fibers, waxes, paraffins, starches, tracantes, cellulose derivatives, polyethylene glycols, silicones, bentonites, silicas, talc or zinc acid may be used as carrier components. Can be.

When the formulation of the present invention is a powder or a spray, lactose, talc, silica, aluminum hydroxide, calcium silicate or polyamide powder may be used, in particular in the case of a spray, additionally chlorofluorohydrocarbon, propane Propellant such as butane or dimethyl ether.

When the formulation of the present invention is a solution or emulsion, a solvent, solvating agent or emulsifying agent is used as the carrier component, such as water, ethanol, isopropanol, ethyl carbonate, ethyl acetate, benzyl alcohol, benzyl benzoate, propylene glycol, 1 Fatty acid esters of, 3-butylglycol oil, glycerol aliphatic ester, polyethylene glycol or sorbitan.

When the dosage form of the present invention is a suspension, liquid carrier diluents such as water, ethanol or propylene glycol, suspending agents such as ethoxylated isostearyl alcohol, polyoxyethylene sorbitol ester and polyoxyethylene sorbitan ester, microcrystalline Cellulose, aluminum metahydroxy, bentonite, agar or tracant and the like can be used.

When the formulation of the present invention is a surfactant-containing cleansing, the carrier component is aliphatic alcohol sulfate, aliphatic alcohol ether sulfate, sulfosuccinic acid monoester, isethionate, imidazolinium derivative, methyltaurate, sarcosinate, fatty acid amide. Ether sulfates, alkylamidobetaines, aliphatic alcohols, fatty acid glycerides, fatty acid diethanolamides, vegetable oils, linolin derivatives or ethoxylated glycerol fatty acid esters and the like can be used.

The present invention also provides a pharmaceutical composition for suppressing or alleviating itching, containing a myrrh extract as an active ingredient.

In the pharmaceutical composition for inhibiting or alleviating the itch of the present invention, the myrrh extract is not toxic to human cells, inhibits IL (interleukin) -31 production and inhibits extracellular signal-regulated kinase (ERK) and NF-kB phosphorylation. It is excellent in effect, and is characterized by an excellent inhibitory or alleviating effect on histamine-independent itch.

In the pharmaceutical composition for suppressing or alleviating the itch of the present invention, the itch may be histamine-independent itch. The histamine-independent itching means itching that is not mediated by histamine.

The pharmaceutical composition for suppressing or alleviating the itch of the present invention may further include a suitable carrier, excipient or diluent commonly used in the preparation of the pharmaceutical composition. Pharmaceutical dosage forms of the compositions according to the invention can be used alone or in combination with other pharmaceutically active compounds, as well as in any suitable collection.

The pharmaceutical compositions according to the invention can be used in the form of powders, granules, tablets, capsules, suspensions, emulsions, syrups, aerosols and the like, oral preparations, external preparations, suppositories, and injections, respectively, according to conventional methods. . Carriers, excipients and diluents that may be included in the pharmaceutical composition comprising the extract include lactose, dextrose, sucrose, sorbitol, mannitol, xylitol, erythritol, maltitol, starch, acacia rubber, alginate, gelatin, calcium phosphate, Various compounds or mixtures including calcium silicate, cellulose, methyl cellulose, microcrystalline cellulose, polyvinyl pyrrolidone, water, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate, mineral oil, and the like. . When formulated, diluents or excipients such as fillers, extenders, binders, wetting agents, disintegrating agents, and surfactants are usually used. Solid preparations for oral administration include tablets, pills, powders, granules, capsules, and the like, which may be added to the extract by mixing at least one excipient such as starch, calcium carbonate, sucrose or lactose, gelatin and the like. It is prepared. In addition to simple excipients, lubricants such as magnesium stearate and talc are also used. Oral liquids include suspensions, liquids, emulsions, and syrups. In addition to water, liquid paraffin, which are commonly used simple diluents, various excipients such as wetting agents, sweeteners, fragrances, and preservatives may be included. have. Formulations for parenteral administration include sterile aqueous solutions, non-aqueous solvents, suspensions, emulsions, lyophilized preparations, suppositories. As the non-aqueous solvent and suspending agent, propylene glycol, polyethylene glycol, vegetable oil such as olive oil, injectable ester such as ethyl oleate and the like can be used. As the base of the suppository, utopsol, macrogol, tween 61, cacao butter, laurin butter, glycerogelatin and the like can be used.

The preferred dosage of the pharmaceutical composition of the present invention depends on the condition and weight of the patient, the extent of the disease, the form of the drug, the route of administration and the duration, and may be appropriately selected by those skilled in the art. The pharmaceutical composition of the present invention can be administered to various mammals such as rats, mice, livestock, humans, and the like. All modes of administration can be expected, for example by oral, rectal or intravenous, intramuscular, subcutaneous, intrauterine dural or intracerebroventricular injection.

Hereinafter, the present invention will be described in detail by way of examples. However, the following Examples are not intended to illustrate the present invention, but the content of the present invention is not limited to the following Examples.

Preparation Example 1 Preparation of Myrrh Extract

100 g of Myrrh (Omni Herb Co., Ltd.) and 2 L of 90% (v / v) ethanol were mixed, left at room temperature for 3 days, filtered using a 0.45 µm filter, and lyophilized at -20 ° C for use in experiments. It was.

Example 1. Cytotoxicity Analysis of Myrrh Extracts

HMC-1 cells (human mast cells) were dispensed in a 96 well culture vessel to a final concentration of 2 × 10 ⁵ / ml and incubated in a 37 ° C., 5% CO ₂ condition for 24 hours. After treatment with myrrh extract (6.25, 12.5, 25, 50 and 100 ㎍ / ㎖) to the cultured cells and incubated for 24 hours again using EZ-cytox Cell Viability Assay Kit (Daeil Biotech, Korea) Cytotoxicity was measured.

As a result, the cytotoxicity was not confirmed in both the untreated control and myrrh extract treatment of the myrrh extract, it can be seen that the myrrh extract of the present invention has little cytotoxicity (Fig. 1).

Example 2 Analysis of Histamine Release Inhibitory Effect

HMC-1 cells were dispensed in a 6 well culture vessel to a final concentration of 5 × 10 ⁵ / ml and incubated in a 37 ° C., 5% CO ₂ condition for 24 hours. After 1 hour pretreatment of myrrh extracts (50 and 100 μg / ml) into cultured cells, 50 nM PMA (phorbl-12-myrsitate-13 acetate) and 1 μM A23187 (calcium ionophore), which induce inflammation (allergic) reactions Stimulated with and incubated for 6 hours, the supernatant was taken and the amount of histamine produced was measured using a Histamine ELISA kit (Enzo Life Sciences, USA).

As a result, it was confirmed that the amount of histamine production was increased in the PMA and A23187 treatment compared to the untreated control, and the high histamine production was confirmed in the pretreatment of myrrh extract as in the PMA and A23187 treatment (Fig. 2). Through this, myrrh extract was found to have no effect on the control of histamine release.

Example 3 Inhibitory Effect of Itching-Related Cytokine IL (interleukin) -31 Production

3-1. PCR (Polymerase chain reaction) analysis

HMC-1 cells were dispensed in a 6 well culture vessel to a final concentration of 5 × 10 ⁵ / ml and incubated in a 37 ° C., 5% CO ₂ condition for 24 hours. Pre-treated myrrh extracts (12.5, 25 and 50 μg / ml) on cultured cells for 1 hour, stimulated with 50 nM PMA and 1 μM A23187, incubated for 2 hours, centrifuged twice with PBS (phosphate-buffered saline) After washing, the RNA was extracted using Ribospin ^™ II (Jinol Biotechnology, Korea), and 5 μg of RNA was quantified by BioSpectrometer® kinetic (Hamgurg, Germany). RNA was then reverse transcribed into First-strand cDNA using PrimeScript ™ 1st strand cDNA Synthesis Kit (TAKARA bio Inc., Japan). Real-time PCR for IL-31 and GAPDH was performed using the Thrmal cycler dice real-time PCR system (TAKARA bio Inc.). The primers used in the experiments were as follows. When real-time PCR was performed, the annealing temperature was set to 60 ° C., and the amplification step was repeated 40 times: IL-31F (5′-TGTGCCAACAGACACCCATG-3 ′). SEQ ID NO: 1), IL-31R (3'-TGTTGGGCTCCAGAGGTCAA-5 '' SEQ ID NO: 2), GAPDH_F (5'-CACTCCTCCACCTTTGACGC-3 '; SEQ ID NO: 3), GAPDH_R (3'-TCCACCACCCTGTTGCTGTA-5'; 4).

As a result, IL-31 expression was increased in the PMA and A23187 treated inflammatory inducers compared to the untreated control, and IL-31 expression was decreased in the group treated with myrrh extract compared to PMA and A23187 treated. It was confirmed (FIG. 3A).

3-2. Western blot analysis

In the same manner as in Example 3-1, myrrh extracts (50 and 100 μg / ml) were pretreated in HMC-1 cells, stimulated with PMA and A23187, incubated for 6 hours, and washed by centrifugation. The washed cells were treated with RIPA buffer (Thermo scientific, USA) to separate the proteins of the cells, and Western blot was performed on the separated proteins to analyze the expression level of IL-31.

As a result, IL-31 expression was increased in the PMA and A23187 treated inflammatory inducing agents compared to the untreated control, and IL-31 expression was dependent on the concentration of the myrrh extract compared to the PMA and A23187 treated groups. It was confirmed that the expression decreases (Fig. 3B).

Example 4. Analysis of phosphorylation inhibitory effect of MAP Kinase (ERK, JNK, p38) and NF-kB

MAP (mitogen-activated protein) kinase is present in the cytoplasm, unphosphorylated, and then phosphorylated and translocated to the nucleus when inflammatory stimuli are applied. MAP kinase induces the activation of transcription factors such as nuclear factor kappa-light-chain-enhancer of activated B cells (NF-kB) and increases the secretion of inflammation-related mediators.

In the same manner as in the Western blot of Example 3-2, the myrrh extract was pretreated with HMC-1 cells, stimulated with PMA and A23187, incubated for 20 minutes, and washed by centrifugation. The washed cells were treated with RIPA buffer to separate the proteins of the cells. Western blots were performed using the isolated proteins to analyze the phosphorylation levels of MAP kinase (ERK, JNK and p38) and NF-kB.

As a result, the phosphorylation of ERK (p-ERK) was increased in the PMA and A23187 treated inflammatory response inducers compared to the untreated control, and in the group pretreated with myrrh extracts (6.25, 12.5, 25 and 50 µg / ml). Phosphorylation of was reduced concentration-dependently. However, the expression of p-JNK and p-p38 was found to be little difference between PMA and A23187 treatment and myrrh extract treatment (Fig. 4A). In addition, phosphorylation of NF-kB (p-NF-kB) was also increased in PMA and A23187 treated groups compared to untreated control, and the phosphorylation level was decreased in the group treated with myrrh extracts (50 and 100 μg / ml). (FIG. 4B).

Example 5 Analysis of Itching Inhibition Effect

Five ICR mice (4 weeks old, males) were placed in a transparent acrylic cage (20 × 26 × 13 cm), 5 mice per experimental group, and allowed to stand for 30 minutes in the same environment. As a control group, physiological saline was administered orally, and the experimental group was administered orally with 50 mg / kg of myrrh extract, 10 mg / kg of antihistamine (cetirizine hydrochloride) and 10 mg / kg of anti-itch standard (prednisolone, prednisolone), respectively. After 60 minutes, itching was induced by subcutaneous injection of compound 48/80 (compound 48/80), which is an itching inducer, at a concentration of 50 μg / site at a height of 0.1 ml between both shoulders of the mouse. Mice injected with the itch inducing agent immediately proceeded for 60 minutes using a micro-camera (ONCCTV, South Korea) according to the method of Mihara et al., 2004, Br J Dermatol, 151 (2), 335-345. Greening was performed by counting the number of scratches of the itch-injecting material in the hind paws by a double blind test.

As a result, the number of scratches was increased in the compound 48/80 treatment group, which was an itch inducing substance, compared to the untreated control group, but in the group pretreated with myrrh extract, antihistamine and prednisolone, the number of scratches was reduced compared to the compound 48/80 treatment group. In particular, the group pretreated with myrrh extract at a concentration of 50 mg / kg was confirmed that the number of scratches was significantly reduced compared to the group pretreated with antihistamine and prednisolone (FIG. 5). Through this, the myrrh extract of the present invention was found to have an excellent itching inhibitory effect compared to the existing products used for itching.

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Claims (6)

Cosmetic composition for inhibiting or alleviating itching, containing myrrh (myrrh) extract as an active ingredient. The cosmetic composition according to claim 1, wherein the extract is extracted using water, a lower alcohol having 1 to 4 carbon atoms, or a mixed solvent thereof. The cosmetic composition of claim 1, wherein the itch is histamine-independent itch. Pharmaceutical composition for suppressing or alleviating itching, containing myrrh (myrrh) extract as an active ingredient. [Claim 5] The pharmaceutical composition of claim 4, wherein the composition has an inhibitory effect on interleukin-31 production and an inhibitory effect of extracellular signal-regulated kinase (ERK) and NF-kB phosphorylation. The method of claim 5, wherein the composition is in the form of powder, granules, tablets, capsules, suspensions, emulsions, syrups, aerosols, external preparations, suppositories, and injections for the suppression or alleviation of itching, characterized in that Pharmaceutical composition.

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