KR20230010167A - Composion of fermented oriental herb, method of manufacturing the same and use of the same - Google Patents

Abstract

The present invention relates to a fermented herbal medicine composition obtained by fermenting a herbal medicine or an extract thereof with a complex strain composed of Bacillus subtilis and lactic acid bacteria, a method for preparing the same, and a composition for anti-cancer, anti-inflammatory or anti-oxidation containing the fermented herbal medicine composition as an active ingredient, the present invention The herbal medicine fermentation composition according to is fermented with a complex strain composed of Bacillus subtilis and lactic acid bacteria, so that the fermentation rate increases compared to the herbal medicine fermentation composition fermented with a single strain of Bacillus subtilis or lactic acid bacteria, thereby increasing the total number of bacteria compared to the same fermentation time, and thus lactic acid bacteria As the number increases, the pH is low, and the activity of proteolytic enzymes or α-amylase, which are products of Bacillus subtilis, can be improved.

Description

Composition of fermented oriental herb, method of manufacturing the same and use of the same}

The present invention relates to a fermented herbal medicine composition, a manufacturing method thereof, and a use thereof, and more particularly, to a fermented herbal medicine composition obtained by fermenting a herbal medicine or an extract thereof with a complex strain composed of Bacillus subtilis and lactic acid bacteria, a method for preparing the same, and a fermented herbal medicine composition as active ingredients It relates to a composition for anti-cancer, anti-inflammatory or antioxidant comprising a.

Fermentation is a process of decomposing organic matter using enzymes possessed by microorganisms to produce substances useful for human life. Representative fermentation microorganisms that produce substances useful for the human body include lactic acid bacteria, yeast, etc. A variety of fermentation products are produced by the fermentation process.

In addition to fermentation, in addition to producing useful substances, lactic acid bacteria or Bacillus subtilis involved in fermentation also play a role in immunity and detoxification in the intestine.

According to ancient medical literature such as Donguibogam and Herbal Medicine, various effects of traditional herbal medicines that have been passed down from ancient times have been reported. There is a growing number of cases where herbal medicine is used to treat incurable diseases that cannot be cured by western medicine.

On the other hand, in the prior art, in fermenting herbal medicines, it is known to prepare herbal medicine fermentation compositions using lactic acid bacteria or Bacillus subtilis alone strain (Patent Document 1, Patent Document 2).

However, as in the prior art, when herbal medicines are fermented using a single strain of lactic acid bacteria or Bacillus subtilis, compared to the case where herbal medicines are fermented using a complex strain, the fermentation rate is lowered, so the total number of bacteria is less compared to the same fermentation time, and accordingly The pH is higher due to the small number of lactic acid bacteria, and the activity of protease or α-amylase, which is a product of Bacillus subtilis, is low.

Republic of Korea Patent Publication No. 10-2012-0103918 Republic of Korea Patent Publication No. 10-2009-0034241

An object of the present invention is to ferment herbal medicines or extracts thereof with a complex strain, so that the fermentation rate increases and the total number of bacteria increases compared to the same fermentation time, thereby increasing the number of lactic acid bacteria and lowering the pH. It is to provide a herbal medicine fermented composition having improved activity such as α-amylase.

Another object of the present invention is to ferment herbal medicines or extracts thereof with complex strains, thereby increasing the fermentation rate and increasing the total number of bacteria compared to the same fermentation time, thereby increasing the number of lactic acid bacteria and lowering the pH, proteolytic enzymes that are products of Bacillus subtilis It is to provide a method for producing a herbal medicine fermented composition having improved activity such as α-amylase or the like.

Another object of the present invention is to provide a use of the herbal medicinal fermented composition for anti-cancer, anti-inflammatory or antioxidant purposes.

In order to achieve the above object, the present invention is astragalus, ginseng, baekchul, bokryeong, jeoryeong, dermis, sukjihwang, saengjihwang, angelica, cnidium, baekjayak, wikmundong, golden, health, contrast and herbal medicines including licorice or extracts of the herbal medicines Provides a herbal medicine fermentation composition fermented with a complex strain consisting of Bacillus subtilis and lactic acid bacteria.

In one embodiment of the present invention, the lactic acid bacteria are Lactobacillus bacteria, Bifidobacterium bacteria, Acetobacter aceti bacteria, Streptococcus thermophilus and Leuconostoc mesenteroides. can be selected from the group consisting of

In one embodiment of the present invention, the bacteria of the genus Lactobacillus are Lactobacillus subtilis, Lactobacillus plantarum, Lactobacillus casei, Lactobacillus acidophilus ) and at least one selected from the group consisting of Lactobacillus lactis.

In one embodiment of the present invention, the Bifidobacterium is 1 selected from the group consisting of Bifidobacterium longum, Bifidobacterium infantis and Bifidobacterium bifidum. There may be more than one species.

In one embodiment of the present invention, the herbal medicine may be included in 55 to 65% by weight based on the total weight of the composition.

In one embodiment of the present invention, the herbal medicine is based on 100 parts by weight of Astragalus, 90 to 110 parts by weight of ginseng, 90 to 110 parts by weight of Baekchul, 90 to 110 parts by weight of Bokryeong, 90 to 110 parts by weight of Jeoryeong, and 90 to 110 parts by weight of dermis. 90-110 parts by weight of Sukjihwang, 90-110 parts by weight of Raw Rehmannia Root, Angelica 90-110 parts by weight, Cnidium 90-110 parts by weight, 90-110 parts by weight of White Peony, 90-110 parts by weight of Maekmundong, 40-60 parts by weight of Golden , 40 to 60 parts by weight of health, 40 to 60 parts by weight of control, and 40 to 60 parts by weight of licorice.

In order to achieve the above object, the present invention is astragalus, ginseng, baekchul, bokryeong, jeoryeong, dermis, sukjihwang, saengjihwang, angelica, cnidium, baekjayak, wikmundong, golden, health, contrast and herbal medicines including licorice or extracts of the herbal medicines It provides a method for producing a herbal medicine fermentation composition comprising the step of fermenting with a complex strain consisting of Bacillus subtilis and lactic acid bacteria.

In one embodiment of the present invention, the lactic acid bacteria are Lactobacillus bacteria, Bifidobacterium bacteria, Acetobacter aceti bacteria, Streptococcus thermophilus and Leuconostoc mesenteroides. can be selected from the group consisting of

In one embodiment of the present invention, the bacteria of the genus Lactobacillus are Lactobacillus subtilis, Lactobacillus plantarum, Lactobacillus casei, Lactobacillus acidophilus ) and at least one selected from the group consisting of Lactobacillus lactis.

In one embodiment of the present invention, the Bifidobacterium is 1 selected from the group consisting of Bifidobacterium longum, Bifidobacterium infantis and Bifidobacterium bifidum. There may be more than one species.

In one embodiment of the present invention, the herbal medicine may be included in 55 to 65% by weight based on the total weight of the composition.

In one embodiment of the present invention, the herbal medicine is based on 100 parts by weight of Astragalus, 90 to 110 parts by weight of ginseng, 90 to 110 parts by weight of Baekchul, 90 to 110 parts by weight of Bokryeong, 90 to 110 parts by weight of Jeoryeong, and 90 to 110 parts by weight of dermis. 90-110 parts by weight of Sukjihwang, 90-110 parts by weight of Raw Rehmannia Root, Angelica 90-110 parts by weight, Cnidium 90-110 parts by weight, 90-110 parts by weight of White Peony, 90-110 parts by weight of Maekmundong, 40-60 parts by weight of Golden , 40 to 60 parts by weight of health, 40 to 60 parts by weight of control, and 40 to 60 parts by weight of licorice may be added.

In one embodiment of the present invention, the fermentation may be incubated for 36 to 96 hours at a temperature of 35 to 39 ° C.

In order to achieve the above object, the present invention provides a pharmaceutical composition for anticancer comprising the herbal composition fermented as an active ingredient.

In order to achieve the above object, the present invention provides a food composition for preventing or improving cancer comprising the herbal medicine fermented composition as an active ingredient.

In order to achieve the above object, the present invention provides an anti-inflammatory pharmaceutical composition comprising the herbal composition fermented as an active ingredient.

In order to achieve the above object, the present invention provides a food composition for preventing or improving inflammation comprising a herbal medicine fermented composition as an active ingredient.

In order to achieve the above object, the present invention provides a cosmetic composition for preventing or improving inflammation comprising the herbal medicine fermented composition as an active ingredient.

In order to achieve the above object, the present invention provides a food composition for antioxidant comprising a herbal medicine fermented composition as an active ingredient.

In order to achieve the above object, the present invention provides a cosmetic composition for antioxidant comprising a herbal medicine fermented composition as an active ingredient.

The fermented herbal medicine according to the present invention is fermented with a complex strain composed of Bacillus subtilis and lactic acid bacteria, so that the fermentation rate is increased compared to the fermented herbal medicine composition fermented with a single strain of Bacillus subtilis or lactic acid bacteria, so that the total number of bacteria compared to the same fermentation time It is possible to provide a fermented herbal medicine composition having a low pH due to an increase in the number of lactic acid bacteria and improved activity of proteolytic enzyme or α-amylase, which are products of Bacillus subtilis.

Figure 1 shows the inhibitory effect of fermented extracts on LPS-induced increase in inflammatory substance production in RAW264.7 cells, where A is NO production, B is iNOS protein expression, and C is COX-2 protein expression. The expression of iNOS and COX-2 was measured by Western blot method, and histogram calculated iNOS or COX-2 expression compared to actin expression (mean ± SD, n = 3 ) was done. *p<0.05, **p<0.01, ***p<0.001 vs. nor (a) or LPS (b)
Figure 2 shows the inhibitory effect on the phosphorylation expression increase of LPS-induced ERK, NF-kB of the fermented extract in RAW264.7 cells, A is ERK protein expression, B is NF-kB protein expression. The expression of ERK, p-ERK, NF-kB, and p-NF-kB was measured by Western blot method, and the histogram was calculated as ERK expression versus p-ERK expression and NF-kB expression versus p-NF-kB expression (mean ±SD, n=3). *p<0.05, **p<0.01, ***p<0.001 vs. nor (a) or LPS (b)
Figure 3 shows the apoptosis inducing effect of fermented extracts in MCF-7 cells, A is Bax, B is cytochrome c, C is cleaved caspase-3, D is PARP, E is Bcl-2, F is ERK protein expression is shown. Each protein expression was measured by the Western blot method, and the histogram was calculated (mean ± SD, n = 3) for the expression of p-ERK compared to actin expression (A to E) or ERK expression (F). *p<0.05, **p<0.01, ***p<0.001 vs. nor (a)
Figure 4 shows the apoptosis inducing effect of the fermented extract in A549 cells, A is Bax, B is cytochrome c, C is cleaved caspase-3, D is PARP, E is Bcl-2, F is ERK protein expression. will be. Expression of each protein was measured by Western blot method, and the histogram was calculated as the expression of p-ERK (F) versus actin expression (A-E) or ERK expression (mean ± SD, n = 3). *p<0.05, **p<0.01, ***p<0.001 vs. nor (a)
Figure 5 shows the cell viability increasing effect of the fermented extract on oxidative damage in HepG2 cells. Data=mean±SD (n=3). **p<0.01, ***p<0.001 vs. nor (a) or H ₂ O ₂ (b)
Figure 6 shows the cell viability increasing effect of the fermented extract on oxidative damage in HepG2 cells. Data=mean±SD (n=3). **p<0.01, ***p<0.001 vs. nor (a) or H ₂ O ₂ (b)

A first embodiment of the present invention is to prepare herbal medicines including Astragalus, ginseng, baekchul, bokryeong, jeoryeong, dermis, sukjihwang, saengjihwang, angelica, cnidium, white peony, wiltongdong, golden, health, contrast and licorice, or extracts of the herbal medicines from Bacillus subtilis And it relates to a herbal medicine fermentation composition fermented with a complex strain consisting of lactic acid bacteria.

Astragalus membranaceus, which is included in the herbal medicine, is a plant belonging to the legume family, and is one of the herbs widely used in oriental medicine. Traditionally, astragalus was mainly used for chronic fatigue, loss of appetite, anemia, wound recovery, fever, Allergy, uterine bleeding, uterine prolapse, etc.

Ginseng (panax ginseng) included in the herbal medicine is dried ginseng root, a plant of the genus Panax ginseng, and in oriental medicine has the effect of greatly reinforcing energy, invigorating the spleen and lungs, quenching thirst and calming the mind, , Modern pharmacologically, it has effects such as central nervous system regulation, immunity enhancement, cardiac heart, and blood sugar lowering.

Baekchul (Atractylodes macrocephala Koidzumi) included in the herbal medicine pharmacologically increases the weight gain and endurance of mice, increases phagocytic ability, promotes cellular immune function, and also regulates intestinal suppression and excitation action, anti-ulcer and It is known to have liver function protection action, immune function enhancement action, vasodilation action, diuretic action, blood sugar lowering action, and anticancer action.

Jeoryeong (Polyporus umbellatus FR.), which is included in the herbal medicine, is a kind of fungal plant generated from the roots of maple, oak, birch, etc., and is a medicinal material widely used in oriental medicine. When there is a mild diuretic effect, it removes edema and causes inflammation to disappear. In addition, it is often applied when there is pain in not being able to urinate due to cystitis or urethritis. It is used for diuretic purposes when edema is severe during pregnancy and when edema is accompanied by beriberi.

The dermis included in the herbal medicine is tangerine peel, which is rich in vitamins and is known as a good medicine for phlegm and cough in winter.

Saengjihwang (rehmannia glutinosa var. purpurea) included in the herbal medicine is a fresh root of Rehmannia glutinosa var. In oriental medicine, it has the effect of lowering heat, generating sound, and quenching thirst, and in modern pharmacology, it has functions such as soothing, anti-inflammatory, enhancing immunity, and hemostasis.

Angelica acutiloba KITAGAWA belonging to the Apiaceae (Umbelliferae) distributed throughout Korea and the roots of its genus plants, its ingredients include ligustilide, butylidene phthal Components such as lead (Butylidene phthalide), cnidilide, isocnilide, and sesquiterpene are known, and therapeutic effects on menstrual irregularities, abdominal pain, and the like are known.

Cnidium officinale included in the herbal medicine is effective in sedation, pain relief, and tonic, and is used to treat headaches, anemia, and gynecological diseases. Collect the rhizomes from September to November, remove the leaves and stems, dry them in the sun, decoct 3 to 6 g, or use them as pills or powders.

In pharmacological experiments, the count included in the herbal medicine shows that the peoniflorin component has sedative action, analgesic action, antipyretic action, anti-inflammatory action, anti-ulcer action, blood pressure lowering action, and coronary vasodilation action, and the phenol component has a sedative action and antipyretic action It is known to exhibit analgesic action, anti-inflammatory action, and hemostatic action.

Liriope platyphylla, which is included in the herbal medicine, is used as a medicinal material in oriental medicine, and is used as an anti-inflammatory, tonic, antitussive, expectorant, and cardiac agent.

Golden (Scutellaria baicalensis) included in the herbal medicine is used as an antipyretic, diuretic, antidiarrheal, choleretic and anti-inflammatory agent in oriental medicine.

The health contained in the herbal medicine promotes secretion of gastric juice, activates intestinal peristalsis, promotes digestion, relieves vomiting, excites the heart, increases blood pressure, and promotes blood circulation. It is known to have anti-inflammatory, analgesic and antibacterial properties.

Contrast (Zizyphi fructus) included in the herbal medicine is known to be effective for anorexia, insomnia, diarrhea, dysentery, gastritis, anemia, ulcer disease, and skin cancer.

The main components of licorice (Glycyrrhiza uralensis Fischer) included in the herbal medicine are sweet saponin, grichilitin, flavonoids, etc., and are effective for muscle pain, cough, expectoration, stomach cramps, raw materials of glycyltin.

In one embodiment of the present invention, the lactic acid bacteria are Lactobacillus bacteria, Bifidobacterium bacteria, Acetobacter aceti bacteria, Streptococcus thermophilus and Leuconostoc mesenteroides. It may be selected from the group consisting of, but is not limited thereto.

In one embodiment of the present invention, the bacteria of the genus Lactobacillus are Lactobacillus subtilis, Lactobacillus plantarum, Lactobacillus casei, Lactobacillus acidophilus ) and at least one selected from the group consisting of Lactobacillus lactis, but is not limited thereto.

In one embodiment of the present invention, the Bifidobacterium is 1 selected from the group consisting of Bifidobacterium longum, Bifidobacterium infantis and Bifidobacterium bifidum. It may be more than one species, but is not limited thereto.

In one embodiment of the present invention, the herbal medicine may be included in 55 to 65% by weight based on the total weight of the composition, but is not limited thereto.

In one embodiment of the present invention, the herbal medicine is based on 100 parts by weight of Astragalus, 90 to 110 parts by weight of ginseng, 90 to 110 parts by weight of Baekchul, 90 to 110 parts by weight of Bokryeong, 90 to 110 parts by weight of Jeoryeong, and 90 to 110 parts by weight of dermis. 90-110 parts by weight of Sukjihwang, 90-110 parts by weight of Raw Rehmannia Root, Angelica 90-110 parts by weight, Cnidium 90-110 parts by weight, 90-110 parts by weight of White Peony, 90-110 parts by weight of Maekmundong, 40-60 parts by weight of Golden , 40 to 60 parts by weight of health, 40 to 60 parts by weight of control, and 40 to 60 parts by weight of licorice, but are not limited thereto.

In one embodiment of the present invention, the composition may have a greater total number of bacteria compared to the fermented herbal medicine composition for the same fermentation time, but is not limited thereto.

In one embodiment of the present invention, the composition may have a lower pH than the herbal medicine fermented composition fermented with Bacillus subtilis or lactic acid bacteria, but is not limited thereto.

In one embodiment of the present invention, the composition may have higher activity of protease and α-amylase than the fermented herbal medicine composition fermented with Bacillus subtilis or lactic acid bacteria, but is not limited thereto.

A second embodiment of the present invention is astragalus, ginseng, baekchul, bokryeong, jeoryeong, dermis, sukjihwang, saengjihwang, angelica, cnidium, white peony, wiltongdong, golden, health, contrast, and herbal medicines including licorice or extracts of the herbal medicines Bacillus subtilis And it provides a method for producing a herbal medicine fermentation composition comprising the step of fermenting with a complex strain consisting of lactic acid bacteria.

In one embodiment of the present invention, the lactic acid bacteria are Lactobacillus bacteria, Bifidobacterium bacteria, Acetobacter aceti bacteria, Streptococcus thermophilus and Leuconostoc mesenteroides. It may be selected from the group consisting of, but is not limited thereto.

In one embodiment of the present invention, the bacteria of the genus Lactobacillus are Lactobacillus subtilis, Lactobacillus plantarum, Lactobacillus casei, Lactobacillus acidophilus ) and at least one selected from the group consisting of Lactobacillus lactis, but is not limited thereto.

In one embodiment of the present invention, the Bifidobacterium is 1 selected from the group consisting of Bifidobacterium longum, Bifidobacterium infantis and Bifidobacterium bifidum. It may be more than one species, but is not limited thereto.

In one embodiment of the present invention, the herbal medicine may be included in 55 to 65% by weight, preferably 58 to 60% by weight, based on the total weight of the composition, but is not limited thereto.

In one embodiment of the present invention, the herbal medicine is 90 to 110 parts by weight of ginseng, preferably 95 to 105 parts by weight, 90 to 110 parts by weight of Baekchul, preferably 95 to 105 parts by weight, Bokryeong, based on 100 parts by weight of Astragalus. 90 to 110 parts by weight, preferably 95 to 105 parts by weight, 90 to 110 parts by weight, preferably 95 to 105 parts by weight, dermis 90 to 110 parts by weight, preferably 95 to 105 parts by weight, 90 to 110 parts by weight 110 parts by weight, preferably 95 to 105 parts by weight, Saenghwang 90 to 110 parts by weight, preferably 95 to 105 parts by weight, Angelica 90 to 110 parts by weight, preferably 95 to 105 parts by weight, Cnidium 90 to 110 parts by weight part, preferably 95 to 105 parts by weight, 90 to 110 parts by weight of white peony, preferably 95 to 105 parts by weight, 90 to 110 parts by weight of Maekmundong, preferably 95 to 105 parts by weight, 40 to 60 parts by weight of gold, preferably 45 to 55 parts by weight, 40 to 60 parts by weight of health, preferably 45 to 55 parts by weight, 40 to 60 parts by weight of control, preferably 45 to 55 parts by weight and 40 to 60 parts by weight of licorice, preferably 45 to 55 parts by weight may be added, but is not limited thereto.

In one embodiment of the present invention, the fermentation may be incubated at a temperature of 35 to 39 ° C, preferably 36 to 38 ° C for 36 to 96 hours, preferably 48 to 72 hours, but is not limited thereto.

In one embodiment of the present invention, the method for producing the herbal medicinal fermented composition is (a) astragalus, ginseng, baekchul, bokryeong, jeoryeong, dermis, sukjihwang, saengjihwang, angelica, cnidium, baekjayak, pulse, gold, health, contrast and licorice Preparing a herbal medicine or an extract of the herbal medicine comprising; (b) fermenting the prepared herbal medicine into a complex strain consisting of Bacillus subtilis and lactic acid bacteria; and (c) freeze-drying the fermented product, but is not limited thereto.

In the first and second embodiments of the present invention, the "extract" refers to an extract obtained by extracting the herbal medicine, a diluted or concentrated solution of the extract, a dried product obtained by drying the extract, and a crude product of the extract. It includes extracts of all formulations that can be formed using extracts themselves and extracts, such as extracts, purified products, or mixtures thereof.

A third embodiment of the present invention relates to an anti-cancer pharmaceutical composition comprising the herbal medicine fermented composition as an active ingredient.

A fourth embodiment of the present invention relates to a food composition for preventing or improving cancer comprising the herbal medicine fermented composition as an active ingredient.

The fifth embodiment of the present invention relates to an anti-inflammatory pharmaceutical composition comprising the herbal medicine fermented composition as an active ingredient.

The sixth embodiment of the present invention relates to a food composition for preventing or improving inflammation comprising a fermented herbal medicine composition as an active ingredient.

The seventh embodiment of the present invention relates to a cosmetic composition for preventing or improving inflammation comprising the herbal medicine fermented composition as an active ingredient.

The eighth embodiment of the present invention relates to a food composition for antioxidant comprising a herbal medicine fermented composition as an active ingredient.

The ninth embodiment of the present invention relates to a cosmetic composition for antioxidation comprising a herbal medicine fermented composition as an active ingredient.

The pharmaceutical composition according to the present invention includes the herbal medicine fermented composition as an active ingredient, and may include a pharmaceutically acceptable carrier.

The pharmaceutically acceptable carrier is one commonly used in formulation and includes, but is not limited to, saline solution, sterile water, Ringer's solution, buffered saline solution, cyclodextrin, dextrose solution, maltodextrin solution, glycerol, ethanol, liposome, etc. It is not, and if necessary, other conventional additives such as antioxidants and buffers may be further included.

In addition, diluents, dispersants, surfactants, binders, lubricants, etc. may be additionally added to formulate formulations for injections such as aqueous solutions, suspensions, emulsions, etc., ophthalmic eye drops, pills, capsules, granules, or tablets. Regarding a suitable pharmaceutically acceptable carrier and formulation, it can be preferably formulated according to each component using the method disclosed in Remington's literature. The pharmaceutical composition of the present invention is not particularly limited in dosage form, but may be formulated as eye drops, injections or oral intake.

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

The pharmaceutical composition according to the present invention is administered in a pharmaceutically effective amount. In the present invention, "pharmaceutically effective amount" means an amount sufficient to treat a disease at a reasonable benefit / risk ratio applicable to medical treatment, and the effective dose level is based on the type, severity, and activity of the drug in the patient. , sensitivity to the drug, time of administration, route of administration and excretion rate, duration of treatment, factors including drugs used concurrently, and other factors well known in the medical field.

The pharmaceutical composition according to the present invention may be administered as an individual therapeutic agent or in combination with other therapeutic agents, may be administered sequentially or simultaneously with conventional therapeutic agents, and may be administered single or multiple times. Considering all of the above factors, it is important to administer an amount that can obtain the maximum effect with the minimum amount without side effects, which can be easily determined by those skilled in the art.

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

The description of the pharmaceutical composition may equally apply to the food composition unless conflicting.

The food composition is a concept including health functional food.

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

The health functional food composition of the present invention is not particularly limited in other ingredients other than containing the above ingredients as essential ingredients in the indicated proportions, and may contain various flavoring agents or natural carbohydrates as additional ingredients like conventional beverages.

Examples of the aforementioned natural carbohydrates include monosaccharides such as glucose, fructose, and the like; disaccharides such as maltose, sucrose and the like; and polysaccharides such as conventional sugars such as dextrins, cyclodextrins, and the like, and sugar alcohols such as xylitol, sorbitol, and erythritol. As flavoring agents other than those mentioned above, natural flavoring agents (thaumatin, stevia extract (eg rebaudioside A, glycyrrhizin, etc.) and synthetic flavoring agents (saccharin, aspartame, etc.) can advantageously be used. The ratio of the natural carbohydrates can be appropriately determined by a person skilled in the art.

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

Ingredients included in the cosmetic composition according to the present invention may include ingredients commonly used in cosmetic compositions in addition to the active ingredients, for example, conventional adjuvants such as stabilizers, solubilizers, vitamins, pigments and fragrances, and It may include a carrier and the like, but is not limited thereto.

The cosmetic composition according to the present invention may be formulated into any formulation known in the art such as softening lotion, nutrient lotion, nutrient cream, moisturizing cream, nutrient essence, gel, lotion, or ointment.

The cosmetic composition according to the present invention, for example, softening lotion, nutrient lotion, nutrient essence, nutrient oil, moisturizing oil, nutrient cream, moisturizing cream, powder, pack, foundation, makeup base, stick, cleansing, shampoo, gel, It can be prepared and used in formulations such as lotions and ointments, and can also be applied in various forms such as liquid, cream, paste, and solid, and can be formulated using conventional cosmetic preparation methods.

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

<Example> (Preparation of herbal medicine fermentation composition)

1. Composition of Oriental Herbal Medicine and Preparation of Extract

The recipe used in this study was prepared by the hot-water extraction method in which distilled water (2.36 L) 10 times the total dry weight was added after measuring the weight of the constituent medicinal materials and extracted with hot water for 180 minutes.

Information on each constituent herbal medicine is described in Table 1 below. The extract was filtered through a 3M paper filter paper, sterilized under pressure at 121° C. and 1.5 atmospheric pressure for 15 minutes, cooled to room temperature, and then used for fermentation.

2. Preparation of fermented extract

The strains used for the fermentation of the far-fetched extract were all obtained from the Biological Resources Center (KCTC: kribb.re.kr) of the Korea Research Institute of Bioscience and Biotechnology (KCTC 3108; Lactoplantibacillus plantanum subsp. plantanum ) and Bacillus subtilis (KCTC 2217: Bacillus subtilis ). used Lactic acid bacteria were subcultured in MRS medium (Difco, Detroit, MI, USA) and Bacillus subtilis were subcultured in NA (Nutrient agar) medium, respectively, and the initial number of bacteria was adjusted to 1×10 ⁶ CFU/ml.

After adjusting the pH of the faraway extract with 1M NaOH, fermentation was inoculated with lactic acid bacteria and Bacillus subtilis at a ratio of 1% (v / v) to the extract capacity, and cultured in an incubator at 37 ° C for 48 hours to prepare a fermented herbal medicine.

The yield of each extract is shown in Table 2 below.

<Experimental example>

1. Anti-inflammatory effect in macrophages

go. Experiment method

① Cell culture

Mouse macrophage RAW 264.7 cells were cultured in Dulbecco's modified eagle's medium (DMEM, Gibco, Carlsbad, CA, USA) medium was used as a culture medium and cultured in an incubator at 37°C and 5% CO ₂ conditions.

② Drug treatment and inflammatory stimulation

RAW 264.7 cells were dispensed in a 60 mm culture dish at 5×10 ⁵ cells/mL, cultured for 24 hours, and then stabilized, treated with each drug at a concentration of 1 mg/mL, and cultured for 1 hour. Thereafter, after treatment with LPS (1 μg/mL) to induce an inflammatory response, the cells were cultured in an incubator at 37° C. and 5% CO ₂ for a certain period of time according to experimental conditions.

③ Nitric oxide (NO) production measurement

After collecting the culture medium from each culture dish treated with drug and LPS, put 100 μL each into a 96-well plate, and add Griess reagent (1% sulfanilamide in 5% phosphoric acid + 1% α-naphthyl amide in H ₂ 0) to 100 Each μL was added and mixed.

After reacting at room temperature for 5 minutes, the degree of color development was measured by absorbance at 540 nm in a microplate reader. The concentration of NO in the culture medium was calculated using a standard quantitative curve of sodium nitrite (NaNO ₂ ).

④ Analysis of inflammatory protein expression changes (Western blot)

RAW 264.7 cells (5×10 ⁵ cells/mL) were dispensed into a 30 mm culture dish and stabilized for 24 hours, treated with each drug (1 mg/mL), and cultured for 1 hour. After treatment with LPS (1 μg/mL) to induce an inflammatory response, the cells were cultured for 15 minutes for ERK protein expression analysis, 1 hour for NF-kB protein expression analysis, and 24 hours for iNOS and COX-2 expression analysis. .

Then, after harvesting each cell, 100 μl of lysis buffer [50 mM HEPES (pH 7.4), 150 mM NaCl, 1% deoxycholate, 1 mM EDTA, 1 mM PMSF, 1 μg/mL aprotinin] was added and incubated at 4°C for 30 minutes. Cell membrane dissolution was induced by the reaction. The protein-containing supernatant was collected by centrifugation at 14,000 rpm for 20 minutes, and the amount of total protein was calculated using Bradford's Protein Assay solution.

For Western blot, 30 μg of the protein isolated from each cell was mixed well with 5× loading buffer, boiled at 95 ° C for 5 minutes to induce protein denaturation, and used for SDS-PAGE (polyacrylamide gel electrophoresis).

After protein electrophoresis, bands on the gel were transferred to a nitrocellulose membrane, stained with Ponceau S Solution, and the membrane was appropriately cut according to the size of each target protein compared to the size marker. After blocking each membrane in 5% skim milk solution for 1 hour to remove non-specific reactions, primary antibodies or control proteins for iNOS, COX-2, ERK, p-ERK, NF-kB, p-NF-kB, respectively It was reacted with the primary antibody of β-lactin at 4°C for 24 hours.

After washing with 1x TBS-T buffer three times for 15 minutes each, it was reacted with a mouse or rabbit secondary antibody conjugated with horseradish peroxidase for 2 hours at room temperature. After washing with 1x TBS-T buffer three times for 15 minutes, the color was developed using ECL solution (BioRad), and the expression level of the band was observed using an automated image analyzer (RAS image system).

The expression level of each target protein was calculated by comparing the image on the computer with the expression level of β-lactin, and presented as a histogram using the results of repeated experiments at least three times.

⑤ Statistical test

All experimental results were expressed as the mean and standard deviation (mean ± SD) for at least three repeated experiments, and statistical significance was determined by one-way ANOVA (Tukey's Test) was used to test, and a case with a p value of 0.05 or less was determined to be significant.

me. Experiment result

Inflammatory response is normally a mechanism by which immune cells recognize external physical or chemical stimuli or bacterial infection and synthesize and secrete various inflammatory mediators to recover or regenerate damaged tissues. , swelling, pain, and other symptoms.

However, excessive inflammation causes various diseases and contributes to changes to chronic diseases, so control (anti-inflammatory) at an appropriate level is required, and the global market size of drugs used to treat various inflammation-related symptoms is It is as big as the anti-cancer drug market.

Nitric oxide (NO) is an important molecule for cell signaling and plays an important role in performing various biological processes as a neurotransmitter in various tissues of the human body (eg, maintaining tension through vasodilation and relaxation).

Inflammatory reactions are induced by synthesizing NO by expressing iNOS (inducible nitric oxide synthase) in immune cells such as macrophages, but excessive production of inflammatory substances causes diseases by increasing abnormal inflammatory reactions. On the other hand, RAW264.7 cells are mouse macrophages that can easily induce an inflammatory response when treated with LPS (lipopolysaccharide), a component of bacterial coat protein, and are therefore used as an experimental model to measure universal anti-inflammatory effects.

(1) Inhibitory effect on NO production and increased expression of iNOS and COX-2

In this study, when RAW264.7 cells were treated with LPS (LPS), NO production increased significantly (p<0.01) compared to the normal group (Nor), which was shown to decrease by each extract treatment.

LPS in all extracts before fermentation (N1, p<0.01), Lactobacillus fermented extract (L1, p<0.01), Bacillus subtilis fermented extract (B1, p<0.01), and Lactobacillus+Bacillus subtilis mixed extract (L+B1, p<0.01) It showed a significant reduction in NO production compared to the group, and in particular, the lactic acid bacteria + Bacillus subtilis mixture extract was found to be the most effective (Fig. 1A).

In addition, the expression of COX-2 (cyclooxygenase-2), a protein that produces NO-producing protein, iNOS, and prostaglandin E2 (PGE2), an inflammatory substance, showed a significant increase in the LPS-treated group compared to the normal group (p<0.01). , p<0.001), which effectively reduced the expression of both iNOS (Figure 1B) and COX-2 (Figure 1C) by each extract treatment.

LPS-stimulated NO production or increased expression of iNOS and COX-2 were effectively reduced in fermented extracts rather than non-fermented extracts, and among fermented extracts, the Lactobacillus + Bacillus subtilis mixed extract was found to be the most effective.

(2) Inhibitory effect on the increase in phosphorylation expression of ERK and NF-kB

Nuclear factor-kappaB (NF-κB), a nuclear transcription factor, is a representative inflammatory regulatory transcription factor, and various pro-inflammatory cytokines (interleukin (IL)-6, IL-1beta, tumor necrosis factor (TNF)- α) production and transcription for the expression of iNOS and COX-2 are induced to increase the secretion of NO and PGE2, which are inflammatory substances.

ERK (extracellular signal-regulated kinase) is a MAPK (mitogen-activated protein kinase) family protein. When phosphorylated and activated by stimulation such as LPS in macrophages, it promotes the expression of transcription factors such as NF-kB, which is necessary for inducing an inflammatory response. It transmits important signals. Therefore, inhibiting the activation of NF-κB and ERK MAPK through phosphorylation is an important advantage in treating inflammatory responses.

In this study, when RAW264.7 cells were treated with LPS, ERK phosphorylation expression (p-ERK) and NF-kB phosphorylation expression (p-NF-kB) were significantly higher (p-NF-kB) compared to the normal group (Nor). <0.01, p<0.001), which decreased by each extract treatment.

First, the increase in phosphorylation expression of ERK by LPS stimulation was more effectively inhibited in the fermented extract than in the non-fermented extract. Bacillus subtilis mixture extracts (L+B1, p<0.001) showed a significant decrease compared to the control group. In particular, the mixed extract of lactic acid bacteria and Bacillus subtilis showed the most effective p-ERK and p-NF-kB inhibitory action (Fig. 2A).

In addition, the increase in phosphorylation expression of NF-kB by LPS stimulation was found in the pre-fermentation extract (N1, p<0.001), Lactobacillus fermentation extract (L1, p<0.01), Bacillus subtilis fermentation extract (B1, p<0.01), and lactic acid bacteria and Bacillus subtilis mixture. All of the extracts (L + B1, p <0.01) were significantly reduced compared to the control group, and, as in ERK, the Lactobacillus + Bacillus subtilis mixture extract was most effectively inhibited (Fig. 2B).

2. Anticancer effect through induction of apoptosis in cancer cells

go. Experiment method

① Cell culture

MCF-7 cells, a human breast cancer cell line, and A549 cells, a lung cancer cell line, were treated with 10% fetal bovine serum (FBS) and 1% penicillin streptomycin (P/S) in Dulbecco's modified Eagle's medium (DMEM, Gibco, Carlsbad, CA, USA) was used and cultured in a CO ₂ incubator at 37°C.

② Drug treatment

After each cancer cell was divided into 5 × 10 ⁵ cells / mL in a 60 mm dish and stabilized by culturing for 24 hours, each extract was treated with a concentration of 2 mg / mL, and 37 ℃, 5% CO ₂ ㅇEinba for 24 hours cultured in an incubator.

③ Analysis of expression changes of apoptotic proteins (Western blot)

After collecting the cancer cells treated with each extract, Lysis buffer was added and reacted at 4 ° C for 30 minutes to induce cell membrane dissolution. The protein-containing supernatant was collected by centrifugation at 14,000 rpm for 20 minutes, and the amount of total protein was measured using Bradford's Protein Assay solution.

After SDS-PAGE of each cell protein (30 μg), bands on the gel were transferred to a nitrocellulose membrane, and membranes for each target protein were prepared. After blocking each membrane with 5% skim milk solution for 1 hour at room temperature, primary antibodies against Bax, Bcl-2, cleaved caspase-3, cytochrome c, PARK, ERK, p-ERK and β-actin and reacted for 24 hours at 4°C.

After washing three times for 15 minutes each with 1x TBS-T buffer, it was reacted with a mouse or rabbit secondary antibody to which horseradish peroxidase was sensitized (conjugated) at room temperature for 2 hours. After washing with 1x TBS-T buffer three times for 15 minutes, the color was developed using an ECL solution, and the expression of the band was observed in a RAS image system.

The expression level of each target protein was calculated in comparison to the expression of β-lactin, and the results of repeated experiments at least three times were presented as a histogram.

④ Statistical test

All experimental results were expressed as mean and standard deviation (mean±SD) for at least three repeated experiments, and statistical significance was determined by one-way ANOVA (Tukey's Test) was used to test, and a case with a p value of 0.05 or less was determined to be significant.

me. Experiment result

Cancer causes various problems by abnormally proliferating cells due to gene mutation, cell growth regulation abnormality, cell death evasion, cell energy metabolism regulation change, and immune system abnormality.

Cancer treatment uses a method of targeting a mechanism that promotes cancer development (recovery and reengineering of a tumor suppressor gene) or inhibiting proliferation by inducing activation of apoptosis mechanism. Among them, the apoptosis induction method is the principle of anticancer drug that has been used for the longest time, and it mainly uses apoptosis-inducing proteins such as bax ((BCL2 Associated X, Apoptosis Regulator), PARP (Poly (ADP-ribose) polymerase), cytochrome c, and caspase-3. by increasing their expression.

Bax plays a variety of roles, including suppression of tumorigenesis, apoptosis of nerve cells during development, maintenance of homeostasis in the lymphatic system and reproductive organs, cell death due to DNA damage, and ischemia-reperfusion injury. can be suppressed. Generally, when apoptosis occurs, Bcl-2 decreases and Bax increases. In addition, when apoptosis occurs, caspases are cleaved, and in particular, caspase-3 is cleaved and converted to cleaved caspase-3, which becomes activated.

Cytochrome c resides in the inner membrane of mitochondria and is released into the cytosol during apoptosis to release calcium, and when caspase 3 is activated, it induces apoptosis by cleavage of PARP, which plays a role in DNA repair. Therefore, increased expression of apoptosis-inducing molecules such as Bax, cytochrome c, caspase-3, and PARP can act favorably on inducing apoptosis in cancer cells.

On the other hand, ERK is also known to be associated with cell growth inhibition and apoptosis induction, as increased expression is observed when cancer cell death is induced.

(1) Apoptosis induction effect in MCF-7 breast cancer cells

In this study, when MCF-7 breast cancer cells were treated with each extract, compared to normal cells (Nor), the pre-fermentation extract (N1), Lactobacillus fermentation extract (L1), Bacillus subtilis fermentation extract (B1), and Lactobacillus + Bacillus subtilis mixed extract ( L+B1, p<0.01) all increased the expression of Bax (Fig. 3A), cytochrome c (Fig. 3B), cleaved caspase-3 (Fig. 3C), and PARP (Fig. 3D) , reduced the expression of Bcl-2 (Fig. 3E). It also increased the phosphorylation expression of ERK (FIG. 3F).

The increase in the expression of these apoptotic proteins in the fermented extract was more effective than in the extract before fermentation, and in particular, the mixed extract of Lactobacillus + Bacillus subtilis was found to be most effective among various fermented extracts.

(2) Apoptosis inducing effect in A549 lung cancer cells

In A549 lung cancer cells, as in MCF-7 cells, compared to normal cells (Nor), all extracts before fermentation (N1), Lactobacillus fermented extract (L1), Bacillus subtilis fermented extract (B1), and Lactobacillus and Bacillus subtilis mixed extract (L+B1) The expression of Bax (FIG. 4A), cytochrome c (FIG. 4B), cleaved caspase-3 (FIG. 4C), and PARP (FIG. 4D) was increased, and Bcl-2 (FIG. 4E ) expression was reduced. It also increased the phosphorylation expression of ERK (FIG. 4F).

In particular, among the various extracts, the mixture extract of Lactobacillus + Bacillus subtilis showed the most excellent apoptosis inducing effect.

This extract is one of the supplements for reinforcing energy and blood in the human body, and is known to be effective in biological defense (immunity), such as increasing the activity of hematopoietic stem cells in the case of bone marrow function deterioration caused by radiation or anticancer drugs, for example. On the other hand, common anti-cancer drugs in both ways allow cells to die by inducing the expression of the above apoptotic proteins as a mechanism (apoptosis) that destroys cancer cells and normal cells.

From this point of view, since this extract has a rather large action to protect cells, it was expected that the action of inducing apoptosis in cancer cells would be small, but rather, it appeared to increase the induction of cell death, so it can exhibit anticancer activity more safely than the side effects of both anticancer drugs. was expected to be In addition, since these effects were significantly increased when fermented than before fermentation, it is considered that it is effective to apply fermentation techniques when applying to cancer patients.

3. Antioxidant effect in hepatocytes

go. Experiment method

① Cell culture

HepG2 cells (liver cells), a human liver cell line, were cultured in DMEM medium containing 10% FBS and 1% P/S as a culture medium at 37°C and 5% CO ₂ in an incubator.

② Drug treatment and stimulation of oxidative damage

HepG2 cells were divided into 5×10 ⁵ cells/mL in a 60 mm dish, cultured for 24 hours to stabilize, treated with each extract (1 mg/mL), and cultured in a CO ₂ incubator for 1 hour. In order to induce oxidative damage, the cells were treated with hydrogen peroxide (H ₂ O ₂ , 200 μM/mL) and cultured for 24 hours.

③ Cell viability measurement

In order to confirm the protective effect of each extract according to the increase in cytotoxicity by H ₂ O ₂ treatment, cell viability was evaluated. First, HepG2 cells (5×10 ⁵ cells/mL) were cultured by dispensing 100 μl per well in a 96-well plate, and then treated with each extract (1 mg/mL) and cultured for 1 hour.

After treatment with H ₂ O ₂ (200 μM/mL) and incubation for 24 hours, 10 μl of a cell viability measurement reagent (EZ-CYTOX, DoGenBio Co., Ltd., Seoul, Korea) was added to the incubator for 1 hour. After the reaction, absorbance (450 nm) was measured in a microplate reader. For quantification of cell viability (%), the viability of each extract treatment group was evaluated relative to the cell viability of normal cells as 100%.

④ Analysis of antioxidant enzyme protein expression (Western blot)

After collecting HepG2 cells treated with each extract, Lysis buffer was added and reacted at 4°C for 30 minutes to induce cell membrane lysis. The protein-containing supernatant was collected by centrifugation at 14,000 rpm for 20 minutes, and the amount of total protein was measured using Bradford's Protein Assay solution.

After SDS-PAGE of each cell protein (30 μg), bands on the gel were transferred to a nitrocellulose membrane, and membranes for each target protein were prepared. After blocking each membrane with 5% skim milk solution for 1 hour at room temperature, it was reacted with primary antibodies against SOD, catalase, HO-1, NRF-2 and β-actin at 4°C for 24 hours. After washing with 1x TBS-T buffer three times for 15 minutes each, it was reacted with a mouse or rabbit secondary antibody to which horseradish peroxidase was sensitized (conjugated) at room temperature for 2 hours. After washing with 1x TBS-T buffer three times for 15 minutes, the color was developed using an ECL solution, and the expression of the band was observed in a RAS image system.

The expression level of each target protein was calculated in comparison to the expression of β-lactin, and the results of repeated experiments at least three times were presented as a histogram.

⑤ Statistical test

All experimental results were expressed as mean and standard deviation (mean±SD) for at least three repeated experiments, and statistical significance was determined by one-way ANOVA (Tukey's Test) was used to test, and a case with a p value of 0.05 or less was determined to be significant.

me. Experiment result

Antioxidant enzymes protect our bodies from diseases by removing reactive oxygen species (ROS). Representative antioxidant enzymes include superoxide dismutase (SOD), catalase (CAT), and glutathione peroxidase (GSH). Among them, SOD is known as the most important enzyme that removes reactive oxygen species first. When SOD encounters active oxygen, it is converted into hydrogen peroxide (H ₂ O ₂ ), and hydrogen peroxide is converted to water through catalase and discharged.

HO-1 (heme oxygenase-1) is an enzyme that acts on the rate-determining step of heme catabolism. corresponds to The expression of HO-1 is regulated by the transcription factor NRF-2 (nuclear factor-2-related factor-2). NRF-2 is a representative factor influencing the induction and expression of antioxidant genes and is expressed in major tissues of the human body. , which is closely related to the induction of expression of HO-1.

(1) protective effect against cytotoxicity

Compared to the normal group (Nor) in the cytotoxicity evaluation, the cell viability was significantly (p<0.001) decreased in the control group treated with H ₂ O ₂ , which was increased by each extract treatment (FIG. 5).

In addition, compared to the control group, the extract before fermentation (N1, p<0.001), the fermented extract of lactic acid bacteria (L1, p<0.001), the fermented extract of Bacillus subtilis (B1, p<0.01), and the mixed extract of lactic acid bacteria and Bacillus subtilis (L+B1, p<0.001) ) All of the treatment groups showed a significant increase in cell viability compared to the control group.

An increase in cell viability was also observed in the extract before fermentation, and no significant difference was observed between each group, suggesting that all extracts have a cell protective effect against oxidative damage caused by H ₂ O ₂ .

(2) Antioxidant enzyme protein expression regulation effect

When each extract was treated with HepG2 hepatocytes in which oxidative damage was induced by H ₂ O ₂ stimulation, the expression of SOD and catalase increased compared to the normal group (Nor). L1), Bacillus subtilis fermented extract (B1) and Lactobacillus + Bacillus subtilis mixed extract treatment decreased (A and B in FIG. 6).

The expression of HO-1 was decreased by oxidative damage, which was significantly increased by the treatment of the lactic acid bacteria fermented extract (L1), the Bacillus subtilis fermented extract (B1), and the lactic acid bacteria + Bacillus subtilis mixed extract, resulting in a higher fermented expression than the non-fermented extract (N1). The extract was found to be more effective in reducing HO-1 expression.

In addition, as in the increased expression of HO-1 (FIG. 6C), a significant decrease was observed in the Bacillus subtilis extract and the Lactobacillus + Bacillus subtilis mixed fermented extract in the expression change of NRF-2 (FIG. 6D).

Therefore, it was confirmed that the expression of antioxidant enzymes was effectively reduced in the fermented extract rather than in the non-fermented extract, especially in the lactic acid bacteria + Bacillus subtilis mixed extract among the fermented extracts.

In many papers, the increase or decrease in the expression of antioxidant enzymes in HepG2 was observed differently depending on the H ₂ O ₂ treatment conditions. It seems that the defense mechanism against oxidative damage by H ₂ O ₂ treatment was activated. The decrease in the expression of these antioxidant enzymes after treatment with the extract is thought to be because the drug maintained a normal state by protecting hepatocytes through scavenging of oxides.

4. Conclusion

(1) In the LPS-induced inflammatory response of RAW264.7 cells, lactic acid bacteria, Bacillus subtilis, and mixed fermented extracts of lactic acid bacteria and Bacillus subtilis all suppressed the expression of iNOS, an inflammatory protein, thereby reducing NO production, inhibiting COX-2 expression and It showed anti-inflammatory effect through inhibition of phosphorylation expression of ERK and NF-kB. It was more effective in fermented extracts than in non-fermented extracts, and among the fermented extracts, it was found to be most effective in lactobacillus + Bacillus subtilis fermented mixed extract.

(2) In MCF-7 breast cancer cells and A549 lung cancer cells, each fermented extract effectively increases the expression of apoptosis-inducing proteins such as Bax, cytochrome c, cleaved caspase-3, and PARP compared to non-fermented extracts, thereby preventing cancer through apoptosis. It was found to be active, especially effective in lactic acid bacteria + Bacillus subtilis mixed fermented extract.

(3) In the H ₂ O ₂ -induced oxidative damage of HepG2 cells, both the non-fermented extract and the fermented extract protected hepatocytes by restoring the decrease in cell viability. It was found that the increase in the expression of antioxidant enzymes such as SOD, catalase, and HO-1 due to oxidative damage could be suppressed through the decrease in expression of . In particular, among the fermented extracts, the mixed fermented extract of Lactobacillus + Bacillus subtilis was found to be very effective in regulating the expression of antioxidant enzymes.

This herbal extract is a drug with anti-inflammatory, anti-cancer and antioxidant effects, and when this extract is fermented (especially Bacillus subtilis or Bacillus subtilis + lactobacillus), the anti-inflammatory, anti-cancer and antioxidant effects are increased, and in particular, a good effect on anti-cancer activity is expected.

<Preparation Example 1> Preparation of pharmaceutical composition

Preparation Example 1-1. manufacture of powders

Herbal medicine fermented composition prepared in Example 1 10 mg

Lactose monohydrate 100 mg

Talc 10 mg

A powder was prepared by mixing the above components and filling in airtight bags.

Preparation Example 1-2. manufacture of tablets

Herbal medicine fermented composition prepared in Example 1 10 mg

Corn Starch 100 mg

Lactose monohydrate 100 mg

Magnesium Stearate 2 mg

After mixing the above ingredients, tablets were prepared by tableting according to a conventional tablet manufacturing method.

Preparation Example 1-3. Manufacture of capsules

Herbal medicine fermented composition prepared in Example 1 10 mg

Microcrystalline Cellulose 3 mg

Lactose monohydrate 14.8 mg

Magnesium Stearate 0.2 mg

After mixing the above components, capsules were prepared by filling gelatin capsules according to a conventional capsule preparation method.

Preparation Example 1-4. Manufacture of injectables

Herbal medicine fermented composition prepared in Example 1 10 mg

Mannitol 180mg

Sterile Distilled Water for Injection 2974 mg

Phosphate Monohydrogen Phosphate 26 mg

After mixing the above components, it was prepared with the above component content per 1 ampoule (2mL) according to a conventional method for preparing injections.

Preparation Example 1-5. Manufacture of Liquids

Herbal medicine fermented composition prepared in Example 1 10 mg

Isomerized sugar 10 mg

Mannitol 5mg

Appropriate amount of purified water

Lemon flavor appropriate amount

The above components are added to and dissolved in purified water according to a conventional manufacturing method, lemon flavor is added in an appropriate amount, purified water is added to adjust the total volume to 100mL, and then sterilized and filled in a brown bottle to prepare a liquid formulation.

<Manufacture Example 2> Manufacture of health food

Preparation Example 2-1. Manufacture of health supplements

Herbal medicine fermented composition prepared in Example 1 10 mg

Appropriate amount of vitamin mixture

Vitamin A Acetate 70 μg

Vitamin E 1.0 mg

Vitamin B1 0.13 mg

Vitamin B2 0.15 mg

Vitamin B6 0.5 mg

Vitamin B12 0.2 μg

Vitamin C 10 mg

10 μg of biotin

Nicotinamide 1.7 mg

Folic acid 50 μg

Calcium Pantothenate 0.5 mg

Appropriate amount of mineral mixture

Ferrous sulfate 1.75 mg

Zinc Oxide 0.82 mg

Magnesium Carbonate 25.3 mg

Potassium Phosphate Monobasic 15 mg

Dibasic Calcium Phosphate 55 mg

Potassium Citrate 30 mg

Calcium Carbonate 100 mg

Magnesium Chloride 24.8 mg

Although the composition ratio of the above vitamin and mineral mixture was mixed with ingredients suitable for relatively healthy food in a preferred embodiment, the mixing ratio may be arbitrarily modified, and the above ingredients are mixed according to the usual health food manufacturing method, and then , Granules can be prepared and used in the preparation of health food compositions according to conventional methods.

Preparation Example 2-2. Manufacture of health drinks

Herbal medicine fermented composition prepared in Example 1 10 mg

15 g of vitamin C

100 g of vitamin E (powder)

19.75 g iron lactate

Zinc oxide 3.5 g

Nicotinamide 3.5 g

Vitamin A 0.2 g

Vitamin B1 0.25 g

Vitamin B2 0.3 g

Quantification of purified water

After mixing the above components according to the usual health drink manufacturing method, stirring and heating at 85 ° C. for about 1 hour, the resulting solution is filtered and obtained in a sterilized 2L container, sealed and sterilized, and then refrigerated according to the present invention. It is used in the manufacture of health beverage compositions.

Although the composition ratio is a mixture of ingredients suitable for a relatively favorite beverage in a preferred embodiment, the mixing ratio may be arbitrarily modified according to regional and ethnic preferences such as the class of demand, the country of demand, and the purpose of use.

As above, specific parts of the content of the present invention have been described in detail, and for those skilled in the art, these specific descriptions are only preferred embodiments, and the scope of the present invention is not limited thereby. It will be clear.

Accordingly, the substantial scope of the present invention will be defined by the appended claims and their equivalents. Simple modifications or changes of the present invention can be easily used by those skilled in the art, and all such modifications or changes can be considered to be included in the scope of the present invention.

Claims (20)

Astragalus, ginseng, baekchul, bokryeong, jeoryeong, dermis, sukjihwang, saengjihwang, angelica, cnidium, baekjayak, macmundong, golden, health, jojo and licorice, or herbal medicines extracts are fermented with complex strains composed of Bacillus subtilis and lactic acid bacteria. Fermented herbal medicine composition.
The method of claim 1, wherein the lactic acid bacteria consist of Lactobacillus, Bifidobacteria, Acetobacter aceti, Streptococcus thermophilus, and Leuconostoc mesenteroides. Herbal medicine fermentation composition, characterized in that selected from the group.
The method of claim 2, wherein the bacteria of the genus Lactobacillus are Lactobacillus subtilis, Lactobacillus plantarum, Lactobacillus casei, Lactobacillus acidophilus And Lactobacillus lactis (Lactobacillus lactis) Chinese herbal medicine fermentation composition, characterized in that at least one selected from the group consisting of.
The method of claim 2, wherein the Bifidobacterium is one selected from the group consisting of Bifidobacterium longum, Bifidobacterium infantis and Bifidobacterium bifidum. Herbal medicine fermentation composition, characterized in that the above.
According to claim 1, wherein the herbal medicine fermented herbal composition, characterized in that contained in 55 to 65% by weight relative to the total weight of the composition.
The method of claim 1, wherein the herbal medicine is based on 100 parts by weight of Astragalus root, 90 to 110 parts by weight of ginseng, 90 to 110 parts by weight of Baekchul, 90 to 110 parts by weight of Bokryeong, 90 to 110 parts by weight of Jeoryeong, and 90 to 110 parts by weight of dermis. , Sukjihwang 90-110 parts by weight, Raw Rehmannia Root 90-110 parts by weight, Angelica 90-110 parts by weight, Cnidium 90-110 parts by weight, White Peony 90-110 parts by weight, Maekmundong 90-110 parts by weight, Golden 40-60 parts by weight, Herbal medicine fermented composition comprising 40 to 60 parts by weight of health, 40 to 60 parts by weight of control and 40 to 60 parts by weight of licorice.
Astragalus, ginseng, baekchul, bokryeong, jeoryeong, dermis, sukjihwang, saengjihwang, angelica, cnidium, baekjayak, macmundong, golden, health, jojo and licorice, or herbal medicines extracts are fermented with complex strains composed of Bacillus subtilis and lactic acid bacteria. Method for producing a herbal medicinal fermented composition comprising the step of
The method of claim 7, wherein the lactic acid bacteria are composed of Lactobacillus, Bifidobacterium, Acetobacter aceti, Streptococcus thermophilus, and Leuconostoc mesenteroides. Method for producing a herbal medicine fermentation composition, characterized in that selected from the group.
The method of claim 8, wherein the bacteria of the genus Lactobacillus are Lactobacillus subtilis, Lactobacillus plantarum, Lactobacillus casei, Lactobacillus acidophilus and Lactobacillus lactis.
The method of claim 8, wherein the Bifidobacterium is one selected from the group consisting of Bifidobacterium longum, Bifidobacterium infantis and Bifidobacterium bifidum. Method for producing a herbal medicine fermented composition characterized in that the above.
The method of claim 7, wherein the herbal medicine is contained in 55 to 65% by weight based on the total weight of the composition.
The method of claim 7, wherein the herbal medicine is based on 100 parts by weight of Astragalus root, 90 to 110 parts by weight of ginseng, 90 to 110 parts by weight of baekchul, 90 to 110 parts by weight of bokryeong, 90 to 110 parts by weight of jeoryeong, and 90 to 110 parts by weight of dermis. , Sukjihwang 90-110 parts by weight, Raw Rehmannia Root 90-110 parts by weight, Angelica 90-110 parts by weight, Cnidium 90-110 parts by weight, White Peony 90-110 parts by weight, Maekmundong 90-110 parts by weight, Golden 40-60 parts by weight, 40 to 60 parts by weight of health, 40 to 60 parts by weight of control, and 40 to 60 parts by weight of licorice are added.
The method of claim 7, wherein the fermentation is performed at a temperature of 35 to 39 ° C. for 36 to 96 hours.
Claims 1 to 6 of any one of herbal medicinal composition according to any one of fermented anti-cancer pharmaceutical composition comprising an active ingredient.
A food composition for preventing or improving cancer, comprising the fermented herbal medicine composition according to any one of claims 1 to 6 as an active ingredient.
An anti-inflammatory pharmaceutical composition comprising the herbal medicine fermented composition according to any one of claims 1 to 6 as an active ingredient.
A food composition for preventing or improving inflammation comprising the fermented herbal medicine composition according to any one of claims 1 to 6 as an active ingredient.
A cosmetic composition for preventing or improving inflammation comprising the herbal medicine fermented composition according to any one of claims 1 to 6 as an active ingredient.
Claims 1 to 6 of any one of herbal medicine fermentation composition according to any one of anti-oxidation food composition containing as an active ingredient.
Claims 1 to 6 of any one of herbal medicinal composition according to any one of fermented antioxidant cosmetic composition containing as an active ingredient.

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