KR102507513B1 - Composition for preventing or treating atopic dermatitis containing natural extract - Google Patents

Abstract

The present invention relates to a composition for preventing or treating atopic dermatitis and psoriasis dermatitis containing a natural extract, and a method for manufacturing the same, wherein the composition has an excellent effect of inhibiting the expression of IL-4, TNF-α, and beta-hexosaminidase, and thus can significantly alleviate itching caused by atopic dermatitis and psoriasis dermatitis, thereby being able to be usefully used for preventing, relieving, and treating atopic dermatitis and psoriasis dermatitis.

Description

Composition for preventing or treating atopic dermatitis or psoriasis dermatitis containing natural extract {Composition for preventing or treating atopic dermatitis containing natural extract}

The present invention relates to a composition for preventing or treating atopic dermatitis or psoriasis dermatitis containing a natural extract and a method for preparing the same.

Atopic dermatitis or psoriasis is caused by genetic, environmental, and immunological causes, and is an allergic disease that becomes more severe in dry climates due to abnormalities in the outermost layer of the skin, which serves as a protective barrier. . Atopic dermatitis and psoriatic dermatitis have a prevalence of about 10 to 20% worldwide, and occur mainly in infants between 1 month and 1 year of age, and are relieved to some extent by the age of 3, but may recur due to living environment, individual differences, etc.

Their main symptoms are severe itching, dry skin, rash, discharge, scab, and scale-like skin (scaly skin), which are usually accompanied by chronic skin inflammation. The patient's scratching behavior caused by itching causes the skin barrier to collapse and causes secondary infection, further exacerbating inflammation. These symptoms can be temporarily alleviated, but may recur and worsen due to stimulants such as the environment and food, and exacerbation and relief are repeated.

Prescriptions for atopic dermatitis or psoriatic dermatitis are mainly drug therapies such as steroids, antihistamines, and antibiotics. Steroids (cortical hormones) have anti-inflammatory and immunosuppressive effects and are highly effective, but long-term application may cause side effects such as skin weakness, systemic hormone symptoms, and addiction. Therefore, there is a need to develop a new composition for the treatment of atopic dermatitis or psoriatic dermatitis that is effective for atopic dermatitis or psoriatic dermatitis and has no side effects.

Republic of Korea Patent Registration No. 10-1828204

The inventors of the present invention completed the present invention by discovering that the therapeutic effect on these dermatitis significantly increases when natural extracts are combined and fermented in a specific ratio during research for the treatment of atopic dermatitis and psoriasis dermatitis.

Accordingly, an object of the present invention is to provide a pharmaceutical composition for preventing or treating atopic dermatitis and psoriasis dermatitis, comprising a fermented product of mixed extracts of cheonnyeoncho, sorrel, purslane, plantain and cockroach as an active ingredient.

Another object of the present invention is to provide a method for preparing the composition.

In order to achieve the object of the present invention, the present invention provides a pharmaceutical composition for preventing or treating atopic dermatitis and psoriasis dermatitis, comprising a fermented product of mixed extracts of cheonnyeoncho, sorrel, purslane, plantain and cockroach as an active ingredient.

In addition, the present invention comprises the steps of a) preparing a mixed extract of Cheonnyeoncho, sorrel, purslane, plantain and cockle;

b) fermenting at 40 to 60 ° C. for 12 hours by adding rice nuruk treated with Hwanggukgyun to the mixed extract of step a);

c) cooling the mixed extract treated in step b) to 25 to 35° C., and then fermenting the mixture at 40 to 60° C. for 15 hours; and

d) centrifuging the mixed extract treated in step c) at 3400 to 3600 rpm for 10 to 20 minutes and then obtaining a supernatant, providing a method for preparing a pharmaceutical composition for preventing or treating atopic dermatitis or psoriasis dermatitis.

In one embodiment of the present invention, the weight ratio of the extracts of cheonnyeoncho, sorrel, purslane, plantain, and cockroach may be 1: 3.5: 1: 1: 1.

As an embodiment of the present invention, the gossip may be a gossip stem.

In one embodiment of the present invention, the composition can inhibit beta-hexosaminidase (β-hexosaminidase), IL-4 and TNF-α expression.

In one embodiment of the present invention, the cheonnyeoncho is cooled to -20 ° C and pre-treated for drying for 36 hours, the plantain is pre-treated for drying for 20 hours, and the cockroach is heated at 100 ° C for 20 minutes and then fumigated The purslane may be pre-treated for drying for 2 hours, the purslane may be pre-treated for drying for 18 hours after fumigation, and the sorrel may be pre-treated for drying for 24 hours.

The composition of the present invention has an excellent effect of inhibiting the expression of IL-4, TNF-α and beta-hexosaminidase, and can significantly alleviate itching caused by atopy and psoriatic dermatitis. can be put to good use.

Figure 1 compares the amount of IL-4 secretion according to treatment with the composition of the present invention.
Figure 2 compares the amount of TNF-α secretion according to the treatment of the composition of the present invention.
Figure 3 is a comparison of beta-hexosaminidase secretion according to the composition of the present invention.
Figure 4 compares the number of scratches according to the treatment with the composition of the present invention.

The inventor of the present invention completed the present invention by discovering that the treatment effect of atopy and psoriasis dermatitis significantly increased when natural extracts were combined and fermented in a specific ratio.

Accordingly, the present invention provides a pharmaceutical composition for preventing or treating atopic dermatitis and psoriasis dermatitis, comprising a fermented product of mixed extracts of cheonnyeoncho, sorrel, purslane, plantain, and cockroach as an active ingredient.

In the present invention, the cheonnyeoncho ( Opuntia humifusa ) means that among cacti of the genus 'Opuntia' native to Korea, wintering is possible in the inland, and those native to Jeju Island are often called baeknyeoncho cacti.

In the present invention, the sorghum ( Rumex crispus L. ) is a perennial plant belonging to the Knobaceae family. It is native to Europe in the northern hemisphere, and spreads to the southern hemisphere and is distributed all over the world. The height of the stem is 30-80cm, it grows straight and is dark green, sometimes tinged with purple.

In the present invention, the purslane ( Portulaca oleracea L. ) is an annual plant belonging to the purslane family. It often grows in fields or roadsides. Its vitality is strong, so it can live for a long time even if it is uprooted.

In the present invention, plantain ( Plantago asiatica L. ) is a perennial plant of the plantain family Lamiaceae. In spring and summer, the young shoots are dug up and eaten as herbs, and in the fall, the seeds are dried in the sun and used as medicine.

In the present invention, the cockroach ( Xanthium strumarium var. japonicum HARA) is an annual plant of the Asteraceae family. Distributed in East Asia, Europe and North America. In the Korean Peninsula, it is a native plant as an alien species.

In the present invention, "active ingredient" means a component that exhibits the desired activity alone or can exhibit activity in combination with a carrier that is not active itself.

In the present invention, "prevention" refers to all activities in which the occurrence or onset of one or more symptoms associated with atopy and psoriatic dermatitis is suppressed or delayed by administration of the pharmaceutical composition of the present invention.

In the present invention, “treatment” refers to all activities that suppress, reduce, or improve symptoms of an individual who exhibits one or more symptoms associated with atopy or psoriatic dermatitis by administering the pharmaceutical composition of the present invention, or previously exhibited. it means. The subject may be a mammal including a human, and is preferably a human.

In the present invention, "administration" means introducing the pharmaceutical composition of the present invention into a subject by any suitable method. The route of administration may be oral or parenteral administration through any general route as long as it can reach the target tissue. Preferably parenteral administration, more preferably it can be applied in the form of topical application by injection.

The pharmaceutical composition of the present invention may further include a pharmaceutically acceptable carrier within a range that does not impair the effects of the present invention. The type of pharmaceutically acceptable carrier that can be used in the present invention is not particularly limited, and any carrier commonly used in the art may be used. Examples of the carrier include saline, sterile water, Ringer's solution, buffered saline, albumin injection solution, lactose, dextrose, sucrose, sorbitol, mannitol, xylitol, erythritol, maltitol, maltodextrin, glycerol, ethanol, and the like. , but is not limited thereto. These carriers may be used alone or in combination of two or more.

The pharmaceutical composition of the present invention may further include pharmaceutically acceptable additives within a range that does not impair the effects of the present invention. Types of pharmaceutically acceptable additives that can be used in the present invention are not particularly limited, and any additives commonly used in the art may be used. The additives may include, for example, fillers, binders, disintegrants, lubricants, suspending agents, solvents, antioxidants, pH adjusters, wetting agents, sweeteners and preservatives, but are not limited thereto. These additives may be used alone or in combination of two or more.

The pharmaceutical composition of the present invention may be in a variety of formulations suitable for parenteral administration. Formulations for parenteral administration include, for example, injections, suppositories, respiratory inhalants, aerosols, ointments, solutions, lotions, patches, powders for application, oils, creams, gels, and the like, preferably injections.

The pharmaceutical composition of the present invention is administered in a pharmaceutically effective amount. In the present invention, "pharmaceutically effective amount" means an amount sufficient to treat a disease with a reasonable benefit / risk ratio applicable to medical treatment, and the level of the effective amount is the patient's condition, weight, sex, age, health It may be determined according to the condition, severity of the disease, sensitivity to the drug, time of administration, route of administration, excretion rate, duration of treatment, factors including concomitantly used drugs and other factors well known in the medical arts. The dosage of the pharmaceutical composition of the present invention can be appropriately selected by a person skilled in the art.

Administration of the pharmaceutical composition of the present invention may be administered once a day, may be administered in several divided doses, or may be administered once in several days.

The pharmaceutical composition of the present invention may be administered alone or in combination with other preventive or therapeutic agents for the prevention and treatment of atopy or psoriatic dermatitis, and may be administered simultaneously or sequentially with other preventive or therapeutic agents. In addition, the pharmaceutical composition of the present invention can be used in combination with methods using surgery, hormone therapy, chemotherapy, and biological response modifiers.

In the present invention, the extract refers to a product such as a liquid component (extract solution) obtained by immersing an extraction target in a solvent and then extracting at room temperature or at elevated temperature for a certain period of time, and a solid component obtained by removing the solvent from the liquid component. can The solvent is one or more solvents selected from the group consisting of water, C ₁ to C ₄ alcohol, hexane, and mixtures thereof, and may preferably be an ethanol extract.

In the present invention, the weight ratio of the cheonnyeoncho, sorghum, purslane, plantain, and cockle extract may be 1: 3.5: 1: 1: 1. In the present invention, beta-hexosaminidase (β-hexosaminidase), IL-4 and TNF-α expression inhibitory ability and itching inhibitory effect was confirmed to be remarkably excellent at the above weight ratio (Experimental Examples 2 to 4).

In the present invention, the gossip may be a gossip stem. The sorrel stem extract of the present invention has excellent atopic and psoriatic dermatitis treatment effects compared to leaf, flower and fruit extracts.

In the present invention, the composition can inhibit beta-hexosaminidase (β-hexosaminidase), IL-4 and TNF-α expression. IL-4 and TNF-α are inflammatory cytokines, and beta-hexosaminidase is an indicator of mast cell degranulation, so it can be seen that the composition of the present invention is effective in treating atopy and psoriasis dermatitis through the effect of inhibiting the expression of these factors. there is.

In another aspect, the present invention provides a method comprising the steps of a) preparing a mixed extract of cheonnyeoncho, sorrel, purslane, plantain and cockle;

b) fermenting at 40 to 60 ° C. for 12 hours by adding rice nuruk treated with Hwanggukgyun to the mixed extract of step a);

c) cooling the mixed extract treated in step b) to 25 to 35° C., and then fermenting the mixture at 40 to 60° C. for 15 hours; and

d) centrifuging the mixed extract treated in step c) at 3400 to 3600 rpm for 10 to 20 minutes and then obtaining a supernatant, providing a method for preparing a pharmaceutical composition for preventing or treating atopic dermatitis or psoriasis dermatitis.

Step a) is a step of preparing a mixed extract, and ethanol may be used as an extraction solvent, preferably immersed in ethanol for 3 days and then extracted. The concentration of the extract is not limited, but may be preferably 200 μg/ml.

Steps b) and c) are fermentation steps of the mixed extract, and effective fermentation is possible when Hwanggukgyun is added in an amount of 0.5 wt% based on the weight of rice when preparing rice nuruk. In addition, the components of the mixed extract can be converted into physiologically active substances through primary and secondary fermentation, and the effect can be maximized within the above temperature and time ranges.

The step d) is a step of separating the active ingredient having an effect of treating atopy and psoriasis, and it is possible to effectively separate the active ingredient within the above rpm and time range.

In the present invention, the cheonnyeoncho is cooled to -20 ° C and then pre-dried for 36 hours, the plantain is pre-dried for 20 hours, and the cockroach is heated at 100 ° C for 20 minutes, then fumigated and pre-treated for 2 hours to dry The purslane may be pre-treated for drying for 18 hours after fumigation, and the sorrel may be pre-treated for drying for 24 hours. Through the pretreatment, it is possible to enhance the atopy and psoriasis treatment effects of the extract.

Hereinafter, a preferred embodiment is presented to aid understanding of the present invention. However, the following examples are provided to more easily understand the present invention, and the content of the present invention is not limited by the following examples.

<Production Example>

Pretreatment of herbal raw materials

Cheonnyeoncho was rapidly cooled at -20 ° C and then dried in a dryer for 36 hours. Plantain was naturally dried in the shade for 20 hours. The cockles were heated in an oven at 100 ° C. for 20 minutes, fumigated three times for 20 minutes each (with methyl bromide), and dried in the shade for 2 hours. Purslane was naturally dried for 18 hours in the shade after 15 minutes of fumigation (broken methyl). Sorikjaeng stems were naturally dried in the shade for 24 hours.

Manufacture of herbal extracts

After cutting the stems of the raspberry into an appropriate size, they were put into an extractor (ASE 300, DIONEX), immersed in 1.5 L of 95% ethanol for 3 days, and then extracted at 50 ° C. for 20 minutes to obtain an extract. Subsequently, the extract was filtered through a paper filter, Watman #2, and dried under reduced pressure. The filtrate obtained was freeze-dried in a freezing dryer (Biotrn) at a cold trap of 70° C. and a chamber temperature of 40° C. for 24 hours to obtain an extract. In addition, extracts of Cheonnyeoncho, purslane, plantain and cockroach were also prepared in the same manner as above.

Rice Nuruk Manufacturing

Non-glutinous rice was washed, soaked for 6 hours, dehydrated, wrapped in cotton cloth, steamed for 30 minutes with a steamer, and steamed for 30 minutes. The steamed rice was spread and cooled to 40 ° C., and dried Aspergillus oryzae was added in an amount of 0.5 wt% relative to the weight of rice and mixed evenly.

Preparation of composition for treatment of atopy and psoriasis

After mixing 1 mg of the Cheonnyeoncho extract, 2.5 mg of purslane extract, 1 mg of plantain extract, and 1 mg of cockle extract, water was added to adjust the concentration of the mixed extract to 200 μg/ml. Then, the rice malt was added and fermented at 50 ° C. for 12 hours. After cooling to 30 ℃ again, it was fermented at 50 ℃ for 15 hours. Then, the supernatant obtained by centrifuging the above fermented product at 3500 rpm for 15 minutes was used as the composition for treating atopy and psoriasis (Sample 1) of the present invention. In addition, samples were additionally prepared with 3 mg (sample 2), 3.5 mg (sample 3), 4 mg (sample 4), and 4.5 mg (sample 5) of only the sorrel stem extract prepared as described above.

<Experimental Example 1: Confirmation of cytotoxicity>

RBL-2H3 (Rat Basophil Leukemia) cells distributed from ATCC (American Type Culture Collection) were cultured in DMEM medium. In order to determine the concentration range to be used in the experiment, the composition of Preparation Example was treated by concentration, and MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltet razolium bromide was prepared using the method of Mosmann (1983). ) analysis was performed.

RBL-2H3 cells were cultured in a 96-well plate with 100 μl of DMEM medium for one day, and then treated with various concentrations (100, 200 and 400 μg/ml) of the composition and cultured for 24 hours. 10 μl of 5 mg/ml MTT solution was added to each well, followed by incubation for 3 hours to induce a reduction reaction, and 100 μl of DMSO (dimethyl sulfoxide) solution was added to completely dissolve purple formazan crystals. dissolved. The degree of color development was measured by absorbance at 570 nm using a spectrophotometer, and the relative cell viability of the sample-treated group was calculated for cytotoxicity based on 100% viability of the untreated group in which only cells were cultured.

Cell viability (% of control) 100 μg/mL 200 μg/mL 400 μg/mL control group 100 100 100 sample 1 98 108 111 sample 2 101 107 112 sample 3 103 109 114 sample 4 99 105 108 sample 5 104 110 114

As shown in Table 1, since no cell damage was observed at various concentrations of all the samples, it can be seen that the composition of the present invention has no cytotoxicity at the above concentrations.

<Experimental Example 2: Confirmation of Inhibition of Inflammatory Cytokine Expression>

In order to measure the concentrations of IL-4 and TNF-α secreted from mast cells, RBL-2H3 cells were seeded in a 24-well plate and cultured for 24 hours. Then, PA (PHORBOL 12-MYRISTATE 13-ACETATE (PMA) 100 pg/ml and Calcium Ionophore A23187 1 μM) was treated, and samples 1 to 5 of the above preparation were treated together (control group was not treated) and cultured for 1 hour while IL -4 and TNF-α were induced. To complete the reaction, it was left on ice for 10 minutes, and the cultured supernatant was centrifuged (1200 rpm, 5 minutes), and then the content was measured using an IL-4 and TNF-α kit (ELISA kit, R&D). .

IL-4 (pg/ml) control group 25.85 sample 1 21.19 sample 2 20.07 sample 3 15.34 sample 4 19.13 sample 5 22.01

TNF-α (pg/ml) control group 33.64 sample 1 28.98 sample 2 26.33 sample 3 13.17 sample 4 19.78 sample 5 24.31

As shown in Tables 2 and 3, it was confirmed that the secretion of IL-4 and TNF-α was significantly inhibited when the composition of the present invention was treated. It was confirmed that there is an ability to inhibit the secretion of -α.

<Experimental Example 3: Confirmation of beta-hexosaminidase expression inhibition ability>

After dispensing 3×10 ⁵ cells per well in a 24-well plate, the RBL-2H3 cells were cultured for 24 hours. Then, PA (PHORBOL 12-MYRISTATE 13-ACETATE (PMA) 100 pg/ml and Calcium Ionophore A23187 1 μM) was treated to induce beta-hexosaminidase production, and samples 1 to 5 of the above preparation were treated together ( The control group was reacted for 1 hour with sample untreated), stopped on ice for 10 minutes, and then stopped after reacting with 1 mM 4-Nitrophenyl N-acetyl-β-Dglucosaminide (0.05M citrate buffer, pH 4.5) substrate for 1 hour. A reagent (0.1M carbonate buffer) was added and the absorbance was measured at 405 nm, and then the secretion reduction rate (%) was calculated for each sample treatment based on the control group.

Beta-hexosaminidase secretion amount (%) control group 100 sample 1 69.11 sample 2 65.28 sample 3 50.52 sample 4 62.78 sample 5 63.81

As shown in Table 4, when the composition of the present invention was treated, the amount of beta-hexosaminidase secretion was significantly inhibited, and in particular, in the case of sample 3, it was confirmed that the secretion inhibition effect was remarkably excellent.

<Experimental Example 4: Confirmation of itching inhibitory effect>

4-week-old male ICR mice reared in a sterile environment were purchased from Central Experimental Animals Co., Ltd., and were acclimatized for one week while supplying enough food and water, and then used for the experiment. The breeding environment was a day and night cycle of 12 hours each, and the temperature (20-22 ℃) and humidity (50-60%) were kept constant.

In order to confirm the anti-pruritic effect of the composition of the present invention, 5 ICR mice per experimental group were placed in a transparent acrylic cage (20×26×13 cm), and left in the same environment for 30 minutes to stabilize. After that, physiological saline was orally administered as a control group, and samples 1 to 5 were orally administered as an experimental group. After 60 minutes, 0.10 mL of compound 48/80 at a concentration of 50 μg/site was subcutaneously injected at the height between both shoulders of the mouse. (s.c.injection) caused itching. The mice injected with the itch-inducing substance were immediately recorded for 60 minutes using a micro-camera according to the method of Mihara, and the number of times they scratched the site where the itch-inducing substance was injected was evaluated with their hind feet.

Number of scratches (number of times/60 minutes) control group 295.30 sample 1 201.09 sample 2 187.55 sample 3 108.61 sample 4 198.67 sample 5 199.33

As shown in Table 5, it was confirmed that the composition of the present invention can significantly suppress itching, and in particular, sample 3 can significantly suppress itching compared to other samples.

Claims (5)

Contains as an active ingredient a fermented product obtained by fermenting a mixed extract obtained by mixing extracts of cheonnyeoncho, sorghum, purslane, plantain, and cockatoo in a weight ratio of 1: 3.5: 1: 1: 1 with rice yeast treated with H.
A pharmaceutical composition for preventing or treating atopic dermatitis or psoriatic dermatitis, which inhibits the expression of beta-hexosaminidase, IL-4 and TNF-α.
delete The pharmaceutical composition according to claim 1, wherein the sobbing is a stem of the sobbing.
delete a) preparing a mixed extract by mixing extracts of cheonnyeoncho, sorrel, purslane, plantain, and cockle at a weight ratio of 1: 3.5: 1: 1: 1;
b) fermenting at 40 to 60 ° C. for 12 hours by adding rice nuruk treated with Hwanggukgyun to the mixed extract of step a);
c) cooling the mixed extract treated in step b) to 25 to 35° C., and then fermenting again at 40 to 60° C. for 15 hours; and
d) centrifuging the mixed extract treated in step c) at 3400 to 3600 rpm for 10 to 20 minutes and obtaining a supernatant, a method for producing a pharmaceutical composition for preventing or treating atopic dermatitis or psoriasis dermatitis.

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