KR20240037489A - Compostions for Improving Intestinal Mircroorganism colonies and Treating Colon Cancer Using an Extract of Cassia Seeds - Google Patents

Abstract

The present invention inhibits cell transformation caused by TPA (12-O-tetradecanoylphorbol-13-acetate), which induces an inflammatory response in normal cells, without showing any particular cytotoxicity to mouse epithelial cells or human keratinocytes, and to mouse It inhibits colon carcinogenesis induced by AOM/DSS in this mouse model, increasing the beneficial microbiome Verrucomicrobia phyum and reducing the harmful microbiome Firmicutes phylum in this mouse model. Disclosed is a composition for improving the intestinal microflora using Cassia chinensis extract and an anti-cancer composition for colon cancer, which not only improves the useful microbiome Akkermansia Genus, but also increases the useful microbiome Akkermansia Genus.
JB6 cells, mouse epithelial cells
HaCaT cells

Description

Composition for improving intestinal microflora using Cassia Seeds extract and anticancer composition for colon cancer {Compostions for Improving Intestinal Microorganism colonies and Treating Colon Cancer Using an Extract of Cassia Seeds}

The present invention relates to a composition for improving the intestinal microbial community using extracts of Cassia chinensis and an anti-cancer composition for colon cancer.

Absence refers to the seeds, leaves, and stems of Cassia obtusifolia L. or Cassia tora L., which belong to the Leguminosae family. Gyeolmyeongja (決明子) is bitter and sweet in taste and cool in nature, and is called by various names such as Yangmyeong (羊明), Cheonri-gwang (千理光), and Chogyeolmyeong (草決明). Herbal medicine is known to have the effect of clearing the liver and clearing the liver and clearing the intestines. In normal cases, it is used when the defects are boiled, ironed, and then consumed as tea. When the head becomes dizzy or painful due to liver fever, the eyes become red, painful, and watery. It is also used to improve eyesight. 『Eastern Pharmacology. 』, it is written that it has the effect of brightening the eyes, making urine flow easily, clearing the liver, and making stool easier, and is said to treat night blindness, hepatitis, and habitual constipation. In the 『Hyangyakdaesan Dictionary』, it is known that among the herbal efficacy of Gyeongmyeongja, Pyeonggan Jamyang maintains the energy of the liver in harmony and has the effect of calming the abnormally proliferating yang (陽邪) (Research on quality standardization of herbal medicine) Project Group, Herbal Medicine Research Division, Ministry of Food and Drug Safety, Gyeol Myeong-ja)

It has been reported that the extract or components of the herbaceous plant have pharmacological antioxidant, anti-inflammatory, and anticancer effects (Pharmacol Rep. 2019 Dec;71(6):1151-1159; Nat Prod Res. 2018 Jun;32(12) ):1476-1480).

Colon cancer is one of the most common cancers and ranks third among all cancer incidence rates, so interest in its treatment is high (2020 release data: National Cancer Information Center, Cancer Information Sharing Center, 2018 National Cancer Registration Statistics). The pathogenesis of colon cancer is known to be intestinal bacteria, pathogens, and chronic enteritis (World J. Gastroenterol. 2008, 14, 3937-3947). The development of colon cancer is also closely related to chronic inflammation, and it has been reported that patients with colitis accompanied by inflammatory bowel disease such as irritable bowel syndrome (IBD) have an increased risk of developing colon cancer (Gastroenterology 2010, 138, 2101-2114 ).

It has been reported that at least 100 trillion microorganisms, 10 times the number of human cells, live in symbiosis in the human gastrointestinal tract (Nature 2010, 464, 59-65). Human intestinal microorganisms are involved in regulating the host's metabolism and immune system, affecting human life and health (S emin. Cancer Biol. 2017, 46, 191-204 ). The greatest diversity of microorganisms resides in the intestines, including Firmicutes , Bacteroidetes , Actinobacteria , Proteobacteria , and Verrucomicrobia at the phylum level. ), etc. are known to exist. Their distribution varies depending on disease status in addition to genetic factors and eating habits. It has been proven that colon cancer patients exhibit an imbalance in the intestinal microflora, with an increase in pathogenic bacteria and harmful metabolites and a decrease in beneficial microorganisms ( Metabolites 2020, 10 , 343 ). In particular, Akkermansia muciniphila , belonging to the Verrucomicrobia phylum, is one of the most abundant species in the human intestinal microflora (0.5-5% of total bacteria), and is one of the most abundant species in the human intestinal microflora (0.5-5% of total bacteria), according to Muriel 2004. Dr. Muriel Derrien isolated it and found that it has a slimming function. Additionally, Akkermansia muciniphila originated from the concept that the human body provides its own “prebiotic” or microbial substrate, i.e., mucinization (D errien et al., 2004; Collado et al., 2007 ), and was found to be specific in the colon. It has been reported that it is produced and decomposed naturally ( Ouwehand et al., 2005 ). It has been reported that lack or reduction of Akkermansia muciniphila is associated with several diseases such as obesity, diabetes, fatty liver, inflammation and cancer ( Nat Rev Gastroenterol Hepatol . 2022 May 31. doi: 10.1038/s41575-022 -00631-9 ).

Therefore Akkermansia Preventing or improving species decline is thought to be useful in the prevention and treatment of colon cancer, and there is a need to develop drugs for this.

Recently, much research is being conducted on anti-cancer disease treatments using herbal medicines that have relatively fewer side effects and are safer than chemical drugs.

In the present invention, the extract of Cassia chinensis is Akkermansia ( Akermansia ) It was completed by confirming that it improves the intestinal microflora by increasing species and has an anticancer effect against colon cancer.

The purpose of the present invention is to provide a composition for improving intestinal microflora using Cassia chinensis extract.

Another object of the present invention is to provide an anticancer composition for colon cancer.

Other objects or specific objects of the present invention will be presented below.

As confirmed in the examples and experimental examples below, the present inventors have found that TPA (12-O-tetradecanoylphorbolol) extract induces an inflammatory response in normal cells without showing any particular cytotoxicity to mouse epithelial cells or human keratinocytes. -13-acetate), inhibits colon cancer induced by AOM/DSS in a mouse model, and promotes the use of the useful microbiome Verrucomicrobia phyum in this mouse model. It was confirmed that it not only increases and reduces the harmful microbiome Firmicutes phylum, but also increases the useful microbiome Akkermansia Genus.

The present invention is provided based on the experimental results described above. In one aspect, the present invention can be viewed as a composition for improving intestinal microflora containing Cassia chinensis extract as an active ingredient, and in another aspect, the present invention can be viewed as a composition for improving intestinal microflora containing Cassia chinensis extract as an active ingredient. It can be identified as an anticancer composition for colon cancer that contains it as an active ingredient.

The composition for improving intestinal microflora of the present invention can be particularly useful for preventing or treating colon cancer, and the anti-cancer composition for colon cancer of the present invention can be usefully used for preventing colon cancer by improving the intestinal microflora.

In this specification, “Cassia extract” means extracting Cassia seeds, leaves, stems, or mixtures thereof with water, lower alcohols with 1 to 4 carbon atoms such as methanol, ethanol, butanol, methylene chloride, ethylene, acetone, hexane, and ether. , extract obtained by leaching using chloroform, ethyl acetate, butylacetate, N,N-dimethylformamide (DMF), dimethyl sulfoxide (DMSO), 1,3-butylene glycol, propylene glycol, or a mixed solvent thereof, It refers to an extract obtained using a supercritical extraction solvent such as carbon dioxide or pentane, or a fraction obtained by fractionating the extract. The extraction method is cold immersion, reflux, heating, ultrasonic radiation, etc., considering the polarity of the active substance, degree of extraction, and degree of preservation. Any method, such as supercritical extraction, may be applied. In the case of a fractionated extract, the extract is suspended in a specific solvent and then mixed with a solvent of different polarity to obtain a fraction obtained by adsorbing the extract on a column filled with silica gel, etc. and then mixed with a hydrophobic solvent, a hydrophilic solvent, or a mixed solvent thereof. It is meant to include fractions obtained using the mobile phase. In addition, the meaning of the extract includes concentrated liquid extract or solid extract from which the extraction solvent has been removed by methods such as freeze-drying, vacuum drying, hot air drying, and spray drying. Preferably, it refers to an extract obtained by using water, ethanol, or a mixed solvent thereof as an extraction solvent, and more preferably, an extract obtained by using a mixed solvent of water and ethanol as an extraction solvent.

Also, in this specification, “intestinal microbial community” refers to the entire intestinal microbiome, which includes Firmicutes , Bacteroidetes , Actinobacteria , and Prodeobacteria at the phylum level. It means including Proteobacteria , Verrucomicrobia , etc.

Also, in this specification, “improving the intestinal microbial community” means increasing useful microorganisms or reducing harmful microorganisms. Specifically, it means increasing microorganisms of the Verrucomicrobia phylum, which are useful microorganisms, especially microorganisms of the Akkermansia genus, or decreasing microorganisms of the Firmicutes phylum, which are harmful microorganisms. .

Also, in this specification, “colon cancer” includes colon cancer that occurs in the colon and rectal cancer that occurs in the rectum, in that the large intestine is divided into the colon and the rectum.

Also, in this specification, “anticancer” means preventing, treating, alleviating, or delaying the onset of cancer.

In addition, in this specification, “active ingredient” refers to an ingredient that exhibits the desired activity alone or can exhibit activity in combination with a carrier that is not active on its own.

In the composition for improving intestinal microflora of the present invention or the anti-cancer composition for colon cancer (hereinafter "composition of the present invention"), the active ingredient is as long as it can exhibit intestinal microflora improving activity and anti-cancer activity against colon cancer, It may be included in any amount (effective amount) depending on the use, formulation, purpose of mixing, etc., and a typical effective amount will be determined within the range of 0.001% by weight to 20.0% by weight based on the total weight of the composition. Here, “effective amount” refers to the intended medical and pharmacological effects, such as the effect of improving intestinal microflora, when the composition of the present invention is administered to mammals, preferably humans, to which it is applied during the administration period recommended by medical experts, etc. It refers to the amount of active ingredient contained in the composition of the present invention. Such effective amounts can be determined experimentally within the scope of the ordinary ability of those skilled in the art.

In addition to the active ingredients, the composition of the present invention further contains any compounds or natural extracts whose safety has already been proven in the art and known to have the relevant activity in order to increase and reinforce the activity of improving intestinal microflora. can do.

These compounds or extracts include compounds, extracts, and pharmaceuticals listed in compendial documents such as the Pharmacopoeia of each country (in Korea, the “Korean Pharmacopoeia”) and the Code of Health Functional Foods of each country (in Korea, it is the “Standards and Specifications for Health Functional Foods” notified by the Ministry of Food and Drug Safety). The laws of each country that govern the manufacture and sale of compounds or extracts and health functional foods that have received product approval in accordance with the laws of each country (in Korea, this is the Pharmaceutical Affairs Act) (in Korea, it is the “Health Functional Foods Act”) 」) includes compounds or extracts whose functionality has been individually recognized. For example, galacto-oligosaccharides, guar gum hydrolyzate, soy oligosaccharides, raffinose, and lactulose powder have been individually recognized as functional for ‘intestinal health (helping to proliferate beneficial bacteria and suppress harmful bacteria in the intestines)’ in accordance with Korea’s Health Functional Foods Act. , indigestible maltodextrin derived from wheat starch, isomaltooligosaccharide, xylo-oligosaccharide, coffee manno-oligosaccharide powder, fructooligosaccharide, etc. may be examples of these compounds or extracts.

In a specific aspect, the composition of the present invention can be viewed as a food composition.

The food composition of the present invention can be manufactured in any form, for example, beverages such as tea, juice, carbonated beverages, and electrolyte drinks, processed oils such as milk and yogurt, gums, rice cakes, Korean snacks, bread, snacks, noodles, etc. It can be manufactured into health functional food preparations such as foods, tablets, capsules, pills, granules, liquids, powders, flakes, pastes, syrups, gels, jellies, bars, etc.

In addition, the food composition of the present invention can have any product classification in terms of legal and functional classification as long as it complies with the enforcement laws and regulations at the time of manufacture and distribution. For example, it is a health functional food according to the Korean 「Act on Health Functional Foods」, or confectionery, beans, tea, and beverages according to each food type according to the food code of the Korean 「Food Sanitation Act」 (Ministry of Food and Drug Safety Notification 「Food Standards and Specifications」) , special purpose food, etc.

The food composition of the present invention may contain food additives in addition to the active ingredients. Food additives can generally be understood as substances that are added to, mixed with, or infiltrated into food when manufacturing, processing, or preserving food. Since they are consumed daily and for a long period of time with food, their safety must be guaranteed. In the Food Additives Code of Laws of each country that regulates the manufacturing and distribution of food (in Korea, it is the Food Sanitation Act), food additives with guaranteed safety are limited in terms of ingredients or functions. In the Korea Food Additive Code (Ministry of Food and Drug Safety Notification “Food Additive Standards and Specifications”), food additives are classified into chemical synthetics, natural additives, and mixed preparations in terms of composition. These food additives are classified into sweeteners and flavors in terms of function. It is classified into preservatives, emulsifiers, acidulants, thickeners, etc.

Sweeteners are used to impart an appropriate sweetness to foods, and either natural or synthetic ones can be used in the composition of the present invention. Preferably, a natural sweetener is used, and natural sweeteners include sugar sweeteners such as corn syrup solids, honey, sucrose, fructose, lactose, and maltose.

Flavoring agents can be used to improve taste or aroma, and both natural and synthetic ones can be used. Preferably, natural products are used. When using natural products, it can serve the purpose of enhancing nutrition in addition to flavor. Natural flavoring agents may be obtained from apples, lemons, tangerines, grapes, strawberries, peaches, etc., or may be obtained from green tea leaves, coriander leaves, bamboo leaves, cinnamon, chrysanthemum leaves, jasmine, etc. You can also use things obtained from ginseng (red ginseng), bamboo shoots, aloe vera, and ginkgo nuts. Natural flavors can be liquid concentrates or solid extracts. In some cases, synthetic flavoring agents may be used, and the synthetic flavoring agents may include esters, alcohols, aldehydes, terpenes, etc.

Preservatives include calcium sodium sorbate, sodium sorbate, potassium sorbate, calcium benzoate, sodium benzoate, potassium benzoate, EDTA (ethylenediaminetetraacetic acid), etc., and emulsifiers include acacia gum, carboxymethyl cellulose, xanthan gum, Examples include pectin, and acidulants include acidic acid, malic acid, fumaric acid, adipic acid, phosphoric acid, gluconic acid, tartaric acid, ascorbic acid, acetic acid, and phosphoric acid. In addition to improving taste, acidulants may be added to ensure that the food composition has an appropriate acidity for the purpose of suppressing the growth of microorganisms.

As a thickening agent, a suspending agent, settling agent, gel forming agent, bulking agent, etc. may be used.

In addition to the food additives described above, the food composition of the present invention may contain bioactive substances or minerals known in the art and whose safety is guaranteed as food additives for the purpose of supplementing and reinforcing functionality and nutrition.

Such physiologically active substances include catechins contained in green tea, vitamins such as vitamin B1, vitamin C, vitamin E, and vitamin B12, tocopherol, dibenzoylthiamine, etc., and minerals include calcium preparations such as calcium citrate and magnesium stearate. Magnesium preparations such as iron citrate, iron preparations such as chromium chloride, potassium iodine, selenium, germanium, vanadium, zinc, etc. are included.

The food composition of the present invention may contain the above-described food additives in an appropriate amount to achieve the purpose of addition depending on the product type.

Regarding other food additives that can be included in the food composition of the present invention, each country's food code or food additive code can be referred to.

The composition of the present invention may be considered a pharmaceutical composition in other specific embodiments.

The pharmaceutical composition of the present invention contains a pharmaceutically acceptable carrier in addition to the active ingredient and can be prepared into an oral formulation or a parenteral formulation depending on the route of administration by a conventional method known in the art. Here, the route of administration may be any suitable route, including topical route, oral route, intravenous route, intramuscular route, and direct absorption through mucosal tissue, and two or more routes may be used in combination. An example of a combination of two or more routes is a case where two or more dosage forms of drugs according to the administration route are combined, for example, when one drug is administered firstly through an intravenous route and the other drug is administered secondarily through a local route.

Pharmaceutically acceptable carriers are well known in the art depending on the route of administration or dosage form, and for specifics, reference can be made to the pharmacopoeia of each country, including the Korean Pharmacopoeia.

When the pharmaceutical composition of the present invention is prepared as an oral dosage form, it can be prepared as powder, granules, tablets, pills, dragees, capsules, solutions, gels, syrups, suspensions, and wafers using a suitable carrier according to methods known in the art. It can be manufactured in a dosage form such as: Examples of suitable carriers include sugars such as lactose, glucose, sucrose, dextrose, sorbitol, mannitol, and xylitol, starches such as corn starch, potato starch, and wheat starch, cellulose, methylcellulose, ethylcellulose, sodium carboxymethylcellulose, Cellulose such as hydroxypropylmethylcellulose, polyvinyl pyrrolidone, water, methylhydroxybenzoate, propylhydroxybenzoate, magnesium stearate, mineral oil, malt, gelatin, talc, polyol, vegetable oil, ethanol, glycerol. etc. can be mentioned. When formulating, appropriate binders, lubricants, disintegrants, colorants, diluents, etc. can be included as needed. Suitable binders include starch, magnesium aluminum silicate, starch paste, gelatin, methylcellulose, sodium carboxymethylcellulose, polyvinylpyrrolidone, glucose, corn sweetener, sodium alginate, polyethylene glycol, and wax. As a lubricant, sodium oleate is used. , sodium stearate, magnesium stearate, sodium benzoate, sodium acetate, sodium chloride, silica, talcum, stearic acid, its magnesium and calcium salts, polydethylene glycol, etc. Disintegrants include starch, methyl cellulose, Examples include agar, bentonite, xanthan gum, starch, alginic acid or its sodium salt. Additionally, diluents include lactose, dextrose, sucrose, mannitol, sorbitol, cellulose, glycine, etc.

When the pharmaceutical composition of the present invention is prepared as a parenteral formulation, it can be formulated in the form of injections, transdermal administration, nasal inhalation, and suppositories along with a suitable carrier according to methods known in the art. When formulated as an injection, an aqueous isotonic solution or suspension can be used as a suitable carrier. Specifically, an isotonic solution such as PBS (phosphate buffered saline) containing triethanolamine, sterile water for injection, or 5% dextrose can be used. . When formulated for transdermal administration, it can be formulated in the form of ointments, creams, lotions, gels, external solutions, paste preparations, linear preparations, and aerol preparations. In the case of nasal inhalation, it can be formulated in the form of an aerosol spray using suitable propellants such as dichlorofluoromethane, trichlorofluoromethane, dichlorotetrafluoroethane, and carbon dioxide. When formulated as a suppository, the carrier is Wethepsol ( witepsol), Tween 61, polyethylene glycols, cocoa fat, laurel paper, polyoxyethylene sorbitan fatty acid esters, polyoxyethylene stearates, sorbitan fatty acid esters, etc. can be used.

Regarding the specific formulation of pharmaceutical compositions, it is known in the art, and references can be made to, for example, Remington's Pharmaceutical Sciences (19th ed., 1995). The above documents are considered a part of this specification.

The preferred dosage of the pharmaceutical composition of the present invention is in the range of 0.001 mg/kg to 10 g/kg per day, preferably 0.001 mg/kg to 1 g, depending on the patient's condition, weight, gender, age, patient's severity, and administration route. It may be in the /kg range. Administration can be done once a day or divided into several times. These dosages should not be construed as limiting the scope of the invention in any respect.

As described above, according to the present invention, it is possible to provide a composition for improving intestinal microflora and an anti-cancer composition for colon cancer using extracts of Cassia chinensis. The composition of the present invention can be commercialized as a functional food or drug.

Figure 1 shows the results showing that the Cassia extract extract did not show cytotoxicity to HaCaT cells.
Figure 2 Results showing the inhibitory effect of TPA-induced JB6 cell transformation of Cassia chinensis extract.
Figure 3 shows the results showing that Cassia chinensis extract inhibits colon cancerization induced by AOM/DSS in a mouse model.
Figure 4 shows that in a mouse model of colon cancer induced by AOM/DSS, Cassia extract increases the useful microbiome Verrucomicrobia phyum and reduces the harmful microbiome Firmicutes phyum. This is a result that shows that.
Figure 5 is a result showing that Cassia extract extract increases the useful microbiome Akkermansia Genus in a mouse model of colon cancer induced by AOM/DSS.
Figure 6 is a result showing that Cassia extract extract increases the useful microbiome Akkermansia uncultured bacterium species in a mouse model of colon cancer induced by AOM/DSS.

The present invention will be described below with reference to examples, but the scope of the present invention is not limited to these examples.

<Example> Effect of improving intestinal microflora and anti-cancer effect on colorectal cancer of extract of Cassia chinensis

1. Materials and experimental methods

1.1 Sample

The extract of Cassia chinensis (seed) used in this experiment was purchased from the Herbal Medicine Resources Research Center of the Korea Institute of Oriental Medicine (resource number 3-20-0125).

The extract was prepared as follows: 10 to 15 times the weight of 70% ethanol was added to the dried powder of Cassia chinensis (extraction part: seed), and extraction was repeated twice for 1 hour at 35 ± 1°C. The yield of the extract was 2.75%. .

The extract sample obtained by freeze-drying was dissolved in dimethylsulfoxide (DMSO) at a concentration of 100 mg/mL.

1.2 Experimental materials

Reagents necessary for cell culture, such as MEM (Minimal Essential Medium), fetal bovine serum (FBS), penicillin/streptomycin (P/S), sodium pyruvate, NEAA (Non-Essential Amino Acid), and 0.25% trypsin, are supplied by GIBCO (Gaithersburg). , MD, USA) 3-(4,5-Dimethyl2-thiazolyl)-2,5-diphenyl-2H-tetrazolium bromide (MTT) was purchased from Sigma-Aldrich Chemical Company (St. Louis, MO, USA). Purchased.

1.3 Cell culture

JB6 cells, which are mouse epithelial cells sensitive to inflammatory factors, were cultured in a 37°C incubator maintained at 5% CO ₂ using MEM containing 5% FBS, 100 U/mL P/S, 1x sodium pyruvate, and 1x NEAA as a medium. did.

1.4 Cell viability assay

HaCaT cells were distributed at ⁷ After adding 10 μL/well of MTT (1 mg/mL) and culturing at 37°C for 1 hour, the medium was removed, and 100 μL of DMSO was dispensed into each well to dissolve formazan. Absorbance was measured at 570 nm using a spectrophotometer (Thermo Fisher Scientific, Vantaa, Finland).

1.5 Anchorage Independent Cell Growth Assay

Dispense 1 mL of 0.33% basal medium Eagle's agar containing 10% FBS into a ⁶ -well plate to make bottom agar, and mix JB6 (8 Then, 3 mL was dispensed onto the bottom agar to make top agar and then solidified at room temperature. After culturing in an incubator at 5% CO ₂ and 37°C for 2 weeks, cell colonies were automatically counted using a computerized microscope system.

1.6 Laboratory animals

As experimental animals, 5-week-old female ICR mice (20-25 g) were purchased from Samtaco Co., Ltd. (Osan, Korea), and the experiment was conducted in a breeding facility equipped with a constant temperature and humidity device, with water and food freely supplied. There was an adaptation period of 1 week before the experiment, normal group, AOM/DSS administration control group, AOM/DSS administration and 20 mg/kg extract administration group, AOM/DSS administration and 40 mg/kg extract administration group, 40 mg/kg extract administration group. The administration group was divided into 5 groups and reared for 12 weeks. Cassia extract was administered orally at 20 or 40 mg/kg/mouse 5 times a week, and after fasting for 24 hours from the end of the experiment, the cervical spine was fractured and laparotomy was performed, and the colon was separated and incised to measure the length and confirm tumor formation. Afterwards, it was placed in a 10% formalin solution and fixed.

This animal experiment was conducted with the approval of Dongshin University's Animal Experiment Ethics Review Committee (approval number: DSU2021-01-05), and the handling and management of experimental animals followed the Animal Experiment Handling Regulations of the Laboratory Animal Ethics Committee.

2. Results

2.1 Treatment of Cassia chinensis extract on normal cells does not show toxicity.

After treating normal cells with doses of 0, 5, 10, 20, and 40 μg/ml of Cassia chinensis extract, cytotoxicity was checked over time. As a result, it was found that even at a high dose of 40 μg/ml, the Cassia chinensis extract did not inhibit cell growth, indicating toxicity. It can be seen that it is not shown (Figure 1).

2.2 Cassia extract inhibits normal cell transformation induced by the inflammatory factor TPA.

As a result of treatment alone with 12-O-tetradecanoylphorbol-13-acetate (TPA), which causes inflammation in normal cells, or in combination with 5, 10, 20, and 40 μg/ml of Cassia chinensis extract, normal cells were transformed (identified as colonies). It was confirmed that this TPA was clearly observed compared to the case without treatment and that it was inhibited in a dose-dependent manner by the Cassia chinensis extract (Figure 2).

2.3 Cassia extract inhibits colon cancerization induced by AOM/DSS

Colon cancerization was confirmed by administering azoxymethane (AOM)/dextran sulfate sodium (DSS), which induce colon cancer, either alone or in combination with 20 or 40 mg/kg of Cassia extract. There was no effect on mouse weight (Figure 3A), and AOM/DSS was observed for disease activity index (DAI), which is an indicator of stool consistency, rectal bleeding, and weight loss. In the administered group, the DAI value was higher than the control group treated only with vehicle, and it was confirmed that the DAI value was decreased when Cassia chinensis extract was administered. That is, the frequency of diarrhea and rectal bleeding was high in the AOM/DSS administration group, but decreased in the Cassia chinensis extract administration group (Figure 3B). In addition, when AOM/DSS is administered, inflammation and polyp occur and the length of the large intestine becomes shorter. When mice were sacrificed after 12 weeks to check the length of the large intestine, the length of the large intestine in the AOM/DSS administered group was shorter than that in the control group. The length of the colon in the group administered Cassia chinensis extract was similar to that in the control group (Figure 3C). The number of polyp, which is an indicator of colorectal cancer, was also found to be higher in the AOM/DSS group than in the control group, and it was confirmed to be reduced in the group administered with Cassia extract extract compared to the AOM/DSS group (Figure 3D).

2.4 In a mouse model of colon cancer induced by AOM/DSS, Cassia extract was found to be useful in the microbiome Verucomicrobia ( Verrucomicrobia ) increases the phyum and harmful microbiome Firmicides ( Firmicutes ) reduce the door

As a result of collecting feces from mice in the control group, AOM/DSS administration group, and AOM/DSS and Cassiasis extract combination administration group and analyzing the microbiome, the beneficial microbiome Verrucomicrobia phylum ( Verrucomicrobia ) was found in the AOM/DSS administration group compared to the control group. phyum) decreased and the harmful microbiome Firmicutes phylum increased, and in the group administered with Cassia extract extract, the useful microbiome Verrucomicrobia phyum increased compared to the AOM/DSS administered group, and the harmful microbiome increased. It was confirmed that the Firmicutes phylum decreased (Figure 4).

2.5 In a mouse model of colon cancer induced by AOM/DSS, Cassia extract extract was used as a useful microbiome of Akkermansia ( Akkermansia ) Increases the genus

As a result of collecting feces from mice in the control group, AOM/DSS administration group, and AOM/DSS combined extract administration group and analyzing the microbiome, the useful microbiome in the AOM/DSS administration group compared to the control group was Akkermansia Genus. ) decreased, and it was confirmed that the beneficial microbiome Akkermansia genus increased in the group administered with Cassia extract extract compared to the group administered with AOM/DSS (FIG. 5).

2.6 In a mouse model of colon cancer induced by AOM/DSS, Cassia extract is useful microbiome Akkermansia-uncultured bacteria ( Akkermansia _Increases uncultured bacterium species

As a result of collecting feces from mice in the control group, AOM/DSS administration group, and AOM/DSS + Cassia extract administration group and analyzing the microbiome, the useful microbiome Akkermansia-uncultured bacteria ( Akkermansia _ uncultured bacterium) was found in the AOM/DSS administration group compared to the control group. ) Species decreased, and it was confirmed that the useful microbiome Akkermansia_uncultured bacterium species increased in the group administered with Cassia extract extract compared to the group administered with AOM/DSS (Figure 6).

Claims (11)

A composition for improving the intestinal microbial community containing Cassia chinensis extract as an active ingredient.
According to paragraph 1,
The composition is characterized in that the extract is a water, ethanol, or mixed solvent extract of Cassiasisiasis seeds.
According to paragraph 1,
A composition that improves the intestinal microbial community by increasing microorganisms of the Verrucomicrobia phylum, which are useful microorganisms, or reducing microorganisms of the Firmicutes phylum, which are harmful microorganisms.
According to paragraph 1,
A composition characterized by improving the intestinal microbiome by increasing microorganisms of the Akkermansia genus.
According to paragraph 1,
The composition is characterized in that it is used for preventing or treating colon cancer.
According to any one of claims 1 to 5,
The composition is characterized in that it is a food composition.
According to any one of claims 1 to 5,
The composition is characterized in that it is a pharmaceutical composition.
An anti-cancer composition for colon cancer containing Cassia chinensis extract as an active ingredient.
According to clause 8,
The composition is characterized in that the extract is a water, ethanol, or mixed solvent extract of Cassiasisiasis seeds.
According to clause 8,
The anti-cancer composition is characterized in that it prevents colon cancer by improving the intestinal microflora.
According to any one of claims 8 to 10,
The composition is characterized in that it is a pharmaceutical composition.