KR102211035B1 - Composition comprising insect extract for treating and improving cancer - Google Patents

Abstract

The present invention relates to a composition for treating and alleviating cancer containing an insect extract. More specifically, the present invention relates to a pharmaceutical composition for treating cancer and a food composition fore alleviating cancer, which contain at least one insect extract among Caelifera, Bombyx mori, Protaetia brevitarsis larvae, and Gryllus bimaculatus, wherein the extract is obtained at room temperature using an aqueous solution containing an antioxidant as an extraction solvent.

Description

Composition for cancer treatment and improvement comprising insect extract {COMPOSITION COMPRISING INSECT EXTRACT FOR TREATING AND IMPROVING CANCER}

The present invention relates to a composition for treating and improving cancer comprising an insect extract, and more particularly, to a composition comprising an extract of an insect obtained at room temperature using an aqueous solution containing an antioxidant as an extraction solvent. It relates to the technology used for the purpose.

In the rapidly changing modern society, many modern people are threatened with health due to various lifestyle-related diseases such as cancer, high blood pressure, angina pectoris, diabetes, obesity, and myocardial infarction due to wrong lifestyle habits. According to statistics from the World Health Organization, cancer is the world's number one mortality rate among lifestyle-related diseases, threatening the lives of modern people, and it is estimated that the mortality rate from cancer will continue to increase in the future.

In particular, the incidence of various lifestyle-related diseases is increasing due to excessive stress, lack of exercise, and changes in meat-oriented eating habits among modern people, and among them, the number of patients dying from colon cancer is increasing rapidly. Colorectal cancer is often unrecognized because the initial symptoms are plain, and many cases have already progressed when discovered.

Despite the development of modern medicine and the development of various medicines, anticancer drugs that have been used up to now have side effects and resistance, so their use has been limited. For example, chemotherapy is widely used in the treatment of colon cancer, but chemotherapy has a side effect of inhibiting immune function by inducing bone marrow damage in patients. As an alternative to the side effects of synthetic drugs, interest in developing anticancer drugs derived from natural products based on pharmacological effects of natural products is increasing. For example, since the intake of food that can increase the patient's immunity can reduce the side effects of chemotherapy and can increase the effectiveness of the recently applied immunotherapy regimen, increasing the patient's immunity is important. It is evaluated as an important factor in enhancing the therapeutic effect.

Meanwhile, as a solution to food shortages caused by population growth and depletion of environmental resources around the world, edible insects are attracting attention as a future food replacement resource. The amount of feed required for insect breeding is not only economical compared to the amount required for livestock breeding, but also the high content of protein in insects is valued nutritionally, so the market for edible insects is expected to grow rapidly. However, there is still insufficient research to use edible insects for medicinal purposes.

In relation to the extraction solvent for obtaining such a natural product extract, for example, in the case of extraction using hot water as disclosed in Korean Patent Laid-Open Patent No. 10-2015-0097891, modification of proteins among the active ingredients may occur. , In the case of using alcohol as an extraction solvent, components with relatively low polarity such as polyphenols are mainly extracted and are not suitable for extracting polar protein components, so extraction that can improve the effect of active ingredients in the extract while being more harmless to the human body If an anticancer ingredient derived from natural products obtained using a solvent can be provided, it is expected to be widely applied in related fields such as medicine and food.

Accordingly, one aspect of the present invention is to provide a pharmaceutical composition for cancer treatment comprising an insect extract obtained by using an aqueous solution containing an antioxidant as an extraction solvent.

Another aspect of the present invention is to provide a food composition for improving cancer comprising an insect extract obtained by using an aqueous solution containing an antioxidant as an extraction solvent.

According to one aspect of the present invention, grasshopper (Caelifera), Baekgangjam ( Bombysis Corpus ), white spotted flower radish ( Protaetia brevitarsis) seulensis ) larvae and twin crickets (Gryllus bimaculatus) containing at least one insect extract, the extract is obtained at room temperature using an aqueous solution containing an antioxidant as an extraction solvent, a pharmaceutical composition for cancer treatment is provided.

According to another aspect of the present invention, grasshopper (Caelifera), Baekgangjam ( Bombysis Corpus ), white spotted flower radish ( Protaetia) brevitarsis seulensis ) larvae and twins cricket (Gryllus bimaculatus) containing at least one insect extract, the extract is obtained at room temperature using an aqueous solution containing an antioxidant as an extraction solvent, cancer improvement food composition is provided.

The insect extract obtained by the present invention can effectively induce the activity of CD4 T cells, CD8 T cells, and dendritic cells in the spleen, and as a result, can induce a reduction in tumor size, so it is widely used as a natural product-based therapeutic agent for anticancer treatment. It is expected to be possible, and furthermore, it is expected to be used as a health functional food that can promote the health of the people.

FIG. 1 shows the results of evaluating the ability to induce surface expression factors on dendritic cells of each of the extracts of Baek Gangnam in Preparation Examples and Comparative Preparation Examples.
Figure 2 shows the results of evaluating the cytokine induction ability to dendritic cells of each of the extracts of the preparation examples and comparative preparation examples.
Figure 3 shows the results of relieving the tumor size of CT-26 colon cancer upon oral administration of the insect extract of the present invention, Figures 3(a) and (b) are comparing the tumor size with the PBS administration group, and Figure 3(c) The tumor volume graph is shown.
Figure 4 shows the immunological activity effect of the insect extract of the present invention in colon cancer transplanted mice, respectively, in the spleen T cell activity (a), CD4 T cell activity (b), and the spleen dendritic cell activity effect ((c) , (d)).

Hereinafter, preferred embodiments of the present invention will be described with reference to the accompanying drawings. However, embodiments of the present invention may be modified in various other forms, and the scope of the present invention is not limited to the embodiments described below.

According to the present invention, a pharmaceutical composition for treating cancer that is safer for the human body is provided as an extract of a natural insect.

In more detail, the composition for cancer treatment of the present invention is a grasshopper (Caelifera), a baekgangjam ( Bombysis Corpus ), a white spotted flower radish ( Protaetia) brevitarsis seulensis ) larvae and twins crickets (Gryllus bimaculatus), including at least one insect extract, the insect extract is obtained at room temperature using an aqueous solution containing an antioxidant as an extraction solvent.

In the present invention, the grasshopper is meant to belong to the locust suborder (Caelifera), for example, may be selected from the rice locust (Oxyachinensis sinuosa) belonging to the locust family. The Baekgang Sleep is a silkworm ( Bombyx mori L.) is a dead body infected with Batrytis bassiana BALS, which is easily crushed and powdered compared to silkworms, and the white spotted flower is an insect belonging to the Coleoptera family, the present invention is the white spotted flower It uses the larva of the plain (larva) as the object of extraction, and the larva of the white-spotted flower is also referred to as'flower'. In addition, the paired cricket refers to an insect of the cricket family of the order Locust Order.

The extract of each insect used in the present invention may be extracted from an aqueous solution at room temperature, at which time the room temperature is from 1° C. to less than 40° C., for example, 4° C. to 37° C., preferably 4° C. to 25° C. .

The water constituting the aqueous solution may be, for example, purified water, and preferably, each insect extract of the present invention is obtained by using an aqueous solution containing 0.01 to 5% by weight of an antioxidant as an extraction solvent, and preferably It was obtained by using an aqueous solution containing 0.1 to 3% by weight of an antioxidant as an extraction solvent.

At this time, the antioxidants that can be used in the present invention include ascorbic acid (vitamin C), disodium ethylenediaminetetracetic acid (EDTA), dicalcium EDTA, tocopherol, butylated hydroxytoluene (BHT), butylated hydroxyanisole (BHA), phosphate and citric acid. It may be one or more selected from the group consisting of, preferably ascorbic acid (ascorbic acid). In particular, ascorbic acid, which can also be referred to as vitamin C, is water-soluble, exists in a reduced form by glutathione, an antioxidant in cells, plays a role in removing active oxygen such as hydrogen peroxide, and can inhibit protein gelation. Therefore, by using ascorbic acid in addition to water as an extraction solvent, it is possible to prevent protein denaturation by inhibiting the oxidation of proteins and fatty acids induced through the formation of free radicals, and it is safer for the human body by removing various free radicals. It can be applied effectively. In addition, for example, a buffer to which phosphate salts are added as an extraction solvent may be used. On the other hand, when a relatively low polarity alcohol such as ethanol is used as the extraction solvent, it is not preferable to extract a polar protein.

When the aqueous solution, which is the extraction solvent of the present invention, contains less than 0.01% by weight of an antioxidant, there is a problem that the antioxidant for preventing denaturation does not function properly.

On the other hand, the insect extract may be in a dry form of the extract, for example, may be in the form of a dry powder. That is, the meaning of the extract of the present invention is to include a dry solid extract.

The process of preparing the insect extract of the present invention includes, for example, immersing the insect in an aqueous solution containing an antioxidant; And obtaining an extract from the aqueous solution in which the insect is immersed.

The water in the immersion step may be an aqueous solution containing 0.01 to 5% by weight of an antioxidant as described above, and the step of immersing the insect in the aqueous solution containing the antioxidant is 1 second to 6 hours, for example It may be 10 seconds to 2 hours, preferably 1 minute to 1 hour, and the immersion step may be performed at room temperature as described above.

Subsequently, a step of obtaining an insect extract from an aqueous solution immersed in insects is performed, and the step is a step of separating a solid and obtaining a liquid extract, for example, centrifuging the aqueous solution immersed in insects to obtain an insect extract. It can be obtained as a supernatant, and in this case, centrifugation can be performed at 3000 to 4000 rpm at room temperature.

Furthermore, it may further include a step of crushing the insect before or after the step of immersing the insect, and when the insect is applied in the form of a powder, extraction efficiency may be further improved.

Further, filtering the insect extract to obtain a filtrate; And it may further include a step of drying the filtrate, impurities may be removed by performing such a filtration step, and further, an extract in a dry form may be obtained by the step of drying, for example An extract in the form of a dry powder can be obtained. The method of performing the filtering step is not particularly limited, and may be performed using a filter, filter paper, or the like.

Meanwhile, the drying step is also not particularly limited, and may be performed by room temperature drying, freeze drying, vacuum drying, hot air drying, spray drying, or the like, preferably freeze drying.

In the pharmaceutical composition for treating cancer and the food composition for improving cancer of the present invention, the insect extract is included as an active ingredient, and in the present invention, "administration" means providing a predetermined substance to the patient by any suitable method, and , The administration route of the composition of the present invention may be administered orally or parenterally through all general routes as long as it can reach the target tissue, and is preferably administered orally.

In the present invention, "patient" refers to a human and all other animals having a disease whose symptoms can be improved by administering the composition of the present invention. The composition according to the invention can be applied not only to humans (treatment, inhibition or prophylaxis), but also to other commercially available animals.

Cancer to which the present invention can be applied may also be referred to as a tumor, meaning a malignant neoplasm, and they are used interchangeably. The type of cancer is not particularly limited, and melanoma, non-small cell lung cancer, small cell lung cancer, lung cancer, liver cancer, retinoblastoma, astrocytoma, glioblastoma, gingival cancer, tongue cancer, leukemia, neuroblastoma, head cancer, cervical cancer , Breast cancer, pancreatic cancer, prostate cancer, kidney cancer, bone cancer, testicular cancer, ovarian cancer, mesothelioma, cervical cancer, gastrointestinal cancer, lymphoma, brain cancer, colon cancer, sarcoma, colon cancer, brain tumor, gastric cancer, esophageal cancer, lymphoma, fibrosarcoma, mast cell tumor And/or bladder cancer.

Further, the cancer includes, for example, mammary gliomas, complex mammary cancer, mammary malignant mixed tumors, intraductal papillary adenocarcinoma, lung adenocarcinoma, squamous cell carcinoma, small cell carcinoma, large cell carcinoma, neuroepithelial tissue tumors such as glioma, ventricular tumor, nerve Cellular tumor, fetal type of extraneurodermal tumor, schwannoma, neurofibroma, meningioma, chronic lymphocytic leukemia, lymphoma, gastrointestinal lymphoma, gastrointestinal lymphoma, small to medium cell type lymphoma, cecal cancer, ascending colon cancer, descending Colon cancer, transverse colon cancer, sigmoid colon cancer, rectal cancer, ovarian epithelial cancer, germ cell tumor, stromal cell tumor, pancreatic duct cancer, invasive pancreatic duct cancer, adenocarcinoma of pancreatic cancer, acinar cell carcinoma, acinar squamous cell carcinoma, giant cell tumor, papillary mucinous tumor in the pancreatic duct , Myxoid cystic adenocarcinoma, pancreatoblastoma, serous cystic adenocarcinoma, solid papillary carcinoma, gastrinoma, glucaconoma, insulinoma, multiple endocrine adenoma 1 (Wermer syndrome), nonfunctional islet cell tumor, somatostatinoma, VIP-producing tumors. However, it is not limited to these.

In particular, cancer to which the present invention can be applied may be classified as a solid cancer or may include cancer that causes metastasis of cancer cells, and preferably, melanoma, brain cancer, lung cancer, gastric cancer, liver cancer, head cancer, cervical cancer, prostate cancer , Pancreatic cancer, colon cancer, and lymphoma. More preferably, it is at least one cancer selected from colon cancer, rectal cancer and colon cancer.

The pharmaceutical composition for cancer treatment of the present invention may contain an insect extract as its active ingredient in any effective amount as long as it can exhibit the improvement activity of cancer intended for treatment depending on the use, formulation, and combination purpose. Herein, "effective amount" refers to the intended medical treatment, such as the treatment or improvement effect of cancer or tumor-related diseases, when the composition of the present invention is administered to a mammal, preferably a human, to which the composition of the present invention is administered during the administration period according to the advice of a medical professional, etc. It refers to the amount of the active ingredient contained in the composition of the present invention, which can exhibit a pharmacological effect.

For example, the pharmaceutical composition of the present invention may contain an insect extract in an amount of 0.01 to 100% by weight based on the total weight of the total pharmaceutical composition, and preferably in an amount of 5 to 70% by weight. will be.

The preferred dosage of the pharmaceutical composition of the present invention is in the range of 0.001 mg/kg to 1 g/kg per day, preferably 1 mg of the insect extract, which is an active ingredient, depending on the patient's condition, weight, sex, age, patient severity, and route of administration. /kg to 500mg/kg, for example, 10 mg/kg to 200mg/kg may be administered. In this case, administration may be performed once a day or divided into several times. However, the dosage may be increased or decreased depending on the route of administration, the severity of the disease, sex, weight, age, and the like.

In addition to the active ingredient, the pharmaceutical composition for treating cancer of the present invention may further contain any compound or natural extract, which has already been verified for safety and is known to have anticancer activity.

In more detail, the pharmaceutical composition of the present invention may be prepared in an oral or parenteral formulation according to an administration route by a conventional method known in the art, including a pharmaceutically acceptable carrier in addition to the active ingredient.

Here, the route of administration may be any suitable route including a local route, an oral route, an intravenous route, an 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 in which two or more formulations of drugs are combined according to the route of administration, for example, when one drug is firstly administered by an intravenous route and secondly, the other drug is administered by a local route.

Pharmaceutically acceptable carriers are well known in the art depending on the route of administration or formulation, and specifically, reference may be made to the pharmacopeias of each country, including the "Korean Pharmacopoeia."

When the pharmaceutical composition of the present invention is prepared in an oral dosage form, powders, granules, tablets, pills, dragees, capsules, solutions, gels, syrups, suspensions, wafers, etc. according to a method known in the art together with a suitable carrier. It can be prepared in the formulation of. Examples of suitable carriers at this time include sugars such as lactose, glucose, sucrose, dextrose, sorbitol, mannitol, xylitol, starches such as corn starch, potato starch, wheat starch, cellulose, methylcellulose, ethylcellulose, sodium carboxymethylcellulose, Celluloses such as hydroxypropylmethylcellulose, polyvinyl pyrrolidone, water, methylhydroxybenzoate, propylhydroxybenzoate, magnesium stearate, mineral oil, malt, gelatin, talc, polyol, vegetable oil, ethanol, grease Serol, etc. are mentioned. In the case of formulation, appropriate binders, lubricants, disintegrants, colorants, and diluents may be included as needed. Suitable binders include starch, magnesium aluminum silicate, starch ferryst, gelatin, methylcellulose, sodium carboxymethylcellulose, polyvinylpyrrolidone, glucose, corn sweetener, sodium alginate, polyethylene glycol, wax, etc., and oleic acid as a lubricant Sodium, sodium stearate, magnesium stearate, sodium benzoate, sodium acetate, sodium chloride, silica, talcum, stearic acid, magnesium salts and calcium salts thereof, polydecylene glycol, etc., and as disintegrating agents starch, methyl cellulose , Agar, bentonite, xanthan gum, starch, alginic acid, or a sodium salt thereof. Moreover, lactose, dextrose, sucrose, mannitol, sorbitol, cellulose, glycine, etc. are mentioned as a diluent.

When the pharmaceutical composition of the present invention is prepared in a parenteral formulation, it may be formulated in the form of an injection, a transdermal administration, a nasal inhalation and a suppository according to a method known in the art together with a suitable carrier. When formulated as an injection, an aqueous isotonic solution or suspension may be used as a suitable carrier, and specifically, triethanol amine-containing PBS (phosphate buffered saline), sterile water for injection, or an isotonic solution such as 5% dextrose may be used. . When formulated into a transdermal dosage form, it can be formulated in the form of an ointment, cream, lotion, gel, external solution, pasta, liniment, air roll, and the like. Nasal inhalants can be formulated in the form of an aerosol spray using a suitable propellant such as dichlorofluoromethane, trichlorofluoromethane, dichlorotetrafluoroethane, and carbon dioxide, and when formulated as a suppository, the carrier is Withepsol ( witepsol), tween 61, polyethylene glycols, cacao butter, laurin paper, polyoxyethylene sorbitan fatty acid esters, polyoxyethylene stearates, sorbitan fatty acid esters, and the like.

The specific formulation of pharmaceutical compositions is known in the art.

On the other hand, according to the present invention, a food composition for improving cancer is provided, and more specifically, one or more of grasshoppers (Caelifera), Baekgangjam ( Bombysis Corpus ), white spotted flower radish ( Protaetia brevitarsis seulensis ) larvae and twin star crickets (Gryllus bimaculatus) It includes an insect extract, wherein the extract is obtained at room temperature using an aqueous solution containing an antioxidant as an extraction solvent, there is provided a food composition for improving cancer.

For example, the food composition of the present invention may contain an insect extract in an amount of 0.01 to 100% by weight, preferably in an amount of 0.5 to 70%, based on the total weight of the food composition. .

With regard to the insect extract, which is an active ingredient in the food composition for improving cancer, the same applies to the contents described in relation to the pharmaceutical composition for treating cancer.

The food composition of the present invention may be prepared in any form, such as beverages such as tea, juice, carbonated beverages, ion beverages, processed oils such as milk and yogurt, gums, rice cakes, Korean sweets, bread, snacks, and noodles. Foods, tablets, capsules, pills, granules, liquids, powders, flakes, pastes, syrup, gels, jelly, can be prepared in health functional food preparations such as bars.

In addition, the food composition of the present invention can be classified as a product as long as it conforms to the enforcement regulations at the time of manufacture and distribution in terms of legal and functional classification. For example, it is a health functional food according to the Korean 「Health Functional Food Act」, or confectionery, bean, tea, and beverages according to each food type according to the food code of the Korean 「Food Sanitation Act」 , Special use 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 food and mixed or infiltrated in manufacturing, processing, or preserving food. Since they are consumed daily and for a long time with food, their safety must be ensured. In the Food Additive Code (“Food Sanitation Act” in Korea) governing the manufacture and distribution of food, food additives with guaranteed safety are limited in terms of ingredients or functions. In the Korean Food Additives Code (“Food Additive Standards and Standards” notified by the Ministry of Food and Drug Safety), food additives are classified into chemical synthetic products, natural additives and mixed preparations in terms of ingredients.These food additives are sweeteners and flavors in terms of function. It is divided into agent, preservative, emulsifier, acidulant, thickener, etc.

Sweeteners are used to impart a suitable sweetness to food, and both natural and synthetic can be used in the composition of the present invention. Preferably, a natural sweetener is used, and examples of the natural sweetener include sugar sweeteners such as corn syrup solids, honey, sucrose, fructose, lactose, and maltose.

Flavoring agents can be used to improve taste or flavor, and both natural and synthetic can be used. Preferably, it is the case of using a natural one. In the case of using natural ingredients, the purpose of nutrient enhancement can be combined in addition to flavor. As a natural flavoring agent, it may be obtained from apples, lemons, tangerines, grapes, strawberries, peaches, etc., or may be obtained from green tea leaves, roundtails, bamboo leaves, cinnamon, chrysanthemum leaves, jasmine, and the like. In addition, you can use those obtained from ginseng (red ginseng), bamboo shoots, aloe vera, and ginkgo. The natural flavoring agent may be a liquid concentrate or a solid extract. In some cases, synthetic flavoring agents may be used, and esters, alcohols, aldehydes, terpenes, and the like may be used as synthetic flavoring agents.

As a preservative, sodium calcium sorbate, sodium sorbate, potassium sorbate, calcium benzoate, sodium benzoate, potassium benzoate, EDTA (ethylenediaminetetraacetic acid), etc. can be used, and as an emulsifier, acacia gum, carboxymethylcellulose, xanthan gum, Pectin, etc. may be mentioned, and as the acidulant, arithmetic, malic acid, fumaric acid, adipic acid, phosphoric acid, gluconic acid, tartaric acid, ascorbic acid, acetic acid, phosphoric acid, and the like can be used. The acidulant may be added so that the food composition has an appropriate acidity for the purpose of suppressing the growth of microorganisms in addition to the purpose of enhancing taste.

As the thickening agent, a suspending agent, a settling agent, a gel-forming agent, a swelling agent, and the like may be used.

In addition to the food additives described above, the food composition of the present invention may include a physiologically active substance or minerals known in the art for the purpose of supplementing and reinforcing functionality and nutritional properties and ensuring stability as a food additive.

In the food composition of the present invention, the food additive as described above may be included in an appropriate amount to achieve the purpose of addition according to the product type. With respect to other food additives that may be included in the food composition of the present invention, reference may be made to the food code of each country or the food additive code.

Hereinafter, the present invention will be described in more detail through specific examples. The following examples are only examples to aid understanding of the present invention, and the scope of the present invention is not limited thereto.

Example

1. Preparation of insect extract

Example : Extraction using aqueous solution containing antioxidant

Edible rice locusts and Gryllus bimaculatus were obtained from S-Worm (Cheonan, Korea), pulverized Baekgangjam powder was obtained from Dainsoap (Cheongju, Korea), and Protaetia brevitarsis seulensis ) larvae (hereinafter referred to as'flowers ') were obtained from E-Double Bugs (Seoul, Korea), respectively.

The flower buds, grasshoppers and twin crickets were each dried and pulverized to prepare a powder.

100 g of each insect powder and 600 mL of 0.16% ascorbic acid in water were mixed, and the mixture was stored at 4° C. for 1 hour. Thereafter, the mixture was centrifuged at 4° C. and 3,000 rpm for 5 minutes to separate and obtain a supernatant. The separated supernatant was filtered under reduced pressure with filter paper (No. 4), and the filtrate was freeze-dried to obtain a dry powder of each insect extract. Each of the dried powders of the thus-obtained Baekanthus extract, grasshopper extract, flower bud extract, and twin cricket extract are referred to as Examples 1, 2, 3, and 4.

Comparative example One: Hydrothermal (Heat) extraction

In the same manner as in Example 1, after preparing the white gill powder, 400 mL of distilled water was added to 20 g, followed by hot water extraction at 90°C for 3 hours. Thereafter, the supernatant was separated by centrifugation at 3500 rpm for 15 minutes, filtered under reduced pressure with filter paper (filter paper, No. 4), and the filtrate was freeze-dried to obtain a dry powder.

Comparative example 2: Hexane (Hexane) extract

In the same manner as in Example 1, after preparing white japonica powder, 400 mL of 99.5% n-hexane was added to 20 g and stirred for 24 hours to extract the fat component. Thereafter, it was filtered under reduced pressure using filter paper (No. 1), and then the n-hexane solvent was removed using a rotary evaporator, and then freeze-dried to obtain a dry powder.

2. Baekgangjam Differentiation of dendritic cells by composition containing extract

Samples were prepared by dissolving 100 mg each of the freeze-dried powders of the white chinensis extract of the present invention obtained in Example 1 and the white chinensis extract obtained by another extraction process obtained in Comparative Example 1 in 30 mL of PBS, and comparison A sample was prepared by dissolving 20 mg of the freeze-dried powder of the extract of Baek chinensis obtained in Example 2 in 10 ml of DMSO. These are referred to as Production Example 1, Comparative Production Example 1 and Comparative Production Example 2, respectively.

Differentiation of dendritic cells using mouse bone marrow-derived cells was performed through GM-CSF/IL-4 (dendritic cell growth factor). In more detail, femoral bone marrow was collected from C57BL/6 mice using bone marrow collection injection, and after washing the collected bone marrow, red blood cells were removed using ammonium chloride. The isolated cells were added to RPMI 1640 (10% FBS, 100 U/ml penicillin/streptomycin, 20 ng/ml GM-CSF, 0.5 ng/ml IL-4) in a 6-well plate for 8 days. Cultured.

3. Evaluation of immune activity of differentiation-induced dendritic cells

(1) Confirmation of surface expression factor

In this experiment, the effects of each component of Preparation Example 1, Comparative Preparation Example 1, and Comparative Preparation Example 2 on surface expression factors, which are active factors of dendritic cells, were investigated. Surface expression factors CD80, CD86, MHC-I and MHC-II are known to be important factors for T cell activity and differentiation, and high expression of these factors can promote T cell activity and differentiation.

Based on these contents, the samples of Preparation Example 1, Comparative Preparation 1, and Comparative Preparation 2 were treated with the dendritic cells cultured for 8 days at a concentration of 100 μg/mL, respectively, for 24 hours.

After 24 hours of treatment as described above, the dendritic cells treated with each sample were anti-CD11c-V450, anti-CD80-FITC, anti-CD86-PE, anti-MHC-I-APC, anti-MHC-II-PE-Cy7 After staining using antibodies, it was analyzed by flow cytometry FACSverse.

As a result, as can be seen in Figure 1, the components extracted by the different extraction methods of Comparative Preparation Example 1 and Comparative Preparation Example 2 from the group treated with the sample of Preparation Example 1 containing the extract of Example 1 of the present invention Compared to that, it was confirmed that the expression of CD80, CD86, MHC-I and MHC-II in dendritic cells was strongly induced.

(2) Cytokine measurement

TNF-alpha, an inflammatory cytokine induced during dendritic cell activation, and IL-12p70, which are important for Th1 polarization of T cells, are known as important indicators for differentiation of Th1-type cells. Therefore, the analysis of the pattern of increase of these indicators can be an important data to predict whether it can control various diseases such as cancer, tuberculosis, and non-tuberculosis mycobacteria.

Based on these contents, the samples of Preparation Example 1, Comparative Preparation 1, and Comparative Preparation 2 were treated with the dendritic cells cultured for 8 days at a concentration of 100 μg/mL, respectively, for 24 hours. Meanwhile, 100 ng/ml of LPS was used as a positive control for the maturation of dendritic cells.

After 24 hours of treatment, TNF-alpha and IL-12p70 in the culture medium were analyzed through ELISA, and as a result, as can be seen in FIG. 2, the sample of Preparation Example 1 containing the extract of Example 1 of the present invention was treated. In the group, it was confirmed that TNF-alpha and IL-12p70 were induced higher than those extracted by the other extraction methods of Comparative Preparation Example 1 and Comparative Preparation Example 2, and Comparative Preparation Example 1 and/or Comparative Preparation Example 2 If the results are not shown, it means that the efficacy was not expressed.

4. Colon cancer tumor size reduction effect Confirm

(1) Experimental animals

Female 7-week-old BALB/c mice were purchased from Orient Bio, allowed to acclimatize for 1 week, and then used for experiments. The temperature of 23±2℃, humidity of 55±5%, and light and dark circulation were acclimated for 1 week in the laboratory animal feedlot maintained every 12 hours, and solid feed and water were supplied freely. Animal experiments for this study were conducted under the approval of the Animal Experiment Ethics Committee of the Advanced Radiation Research Institute of the Korea Atomic Energy Research Institute.

(2) extract administration

In the same manner as in Preparation Example 1, samples of Preparation Examples 2 to 4 were prepared by dissolving the freeze-dried powder of the extract of Baek chinensis of the present invention obtained in Examples 2 to 4 in 30 mL of PBS at 100 mg each. I did.

100 mg/ of each of the thus-obtained Baekanthera extract (BBPE, Preparation Example 1), locust extract (CAPE, Preparation Example 2), Kaleidopsis extract (PBSPE, Preparation Example 3) and twin cricket extract (GBPE, Preparation Example 4) As a result of administering three times a week for 25 days at a dose of kg, as can be seen in FIG. 3, there was no change in tumor volume in the untreated group, but in the GBPE, BBPE, and PBSPE extract-administered groups, tumor administration 14 days, 16 days, It was confirmed that the tumor volume decreased significantly on the 21st, 23rd, and 25th days, and the CAPE extract-administered group showed a significant decrease in the tumor volume on the 21st day.

5. CD4 in the spleen ^- , CD8 T cell And check the effect of dendritic cell activity

In order to analyze whether the reduction in colon cancer tumor size due to oral administration of the insect extract in the above 4 is due to the immune activity effect of the insect extract, after 26 days of colon cancer cell transplantation, the spleen of the mouse was isolated, CD4 T cells, CD8 The activity of T cells and dendritic cells was measured.

As a result of the analysis, as can be seen in FIG. 4, the activity of cytotoxic T cells (CD8 T cells) playing a decisive role in anticancer immunity (CD8 ⁺ CD44 ⁺ ) and the activity of CD4 T cells (CD4 ⁺ CD44 ⁺ ) were normal. Compared to the tumor transplant group, it was confirmed that the activities of cytotoxic T cells and CD4 T cells were significantly increased in the CFPE, GBPE, BBPE, and PBSPE administration groups (Figs. 4(a), (b)).

In addition, as a result of evaluating the activity of dendritic cells, which are innate immune cells that induce T cell activity, it was confirmed that the activity of dendritic cells was effectively induced in the group administered with CFPE, GBPE, BBPE, and PBSPE (Fig. 4(c)). ), (d)).

Although the embodiments of the present invention have been described in detail above, the scope of the present invention is not limited thereto, and various modifications and variations are possible without departing from the technical spirit of the present invention described in the claims. It will be obvious to those of ordinary skill in the field.

Claims (8)

Grasshopper (Caelifera), Baekgangjam ( Bombysis Corpus ), white-spotted flower radish ( Protaetia brevitarsis seulensis ) larvae and twin- star cricket (Gryllus bimaculatus) contains an insect extract of at least one,
The insect extract is obtained at room temperature using an aqueous solution containing ascorbic acid as an extraction solvent, a pharmaceutical composition for cancer treatment.
According to claim 1, The ascorbic acid (ascorbic acid) is contained in an amount of 0.01 to 5% by weight based on the total weight of the aqueous solution, a pharmaceutical composition for cancer treatment.
delete The pharmaceutical composition for cancer treatment according to claim 1, wherein the cancer is at least one cancer selected from colon cancer, rectal cancer and colon cancer.
The pharmaceutical composition for cancer treatment according to claim 1, wherein the pharmaceutical composition is an oral dosage form.
The pharmaceutical composition for cancer treatment according to claim 1, wherein the pharmaceutical composition comprises an insect extract in an amount of 0.01 to 100% by weight based on the total weight of the total pharmaceutical composition.
The pharmaceutical composition for cancer treatment according to claim 1, wherein the pharmaceutical composition has an oral dosage in the range of 0.001mg/kg to 1g/kg per day.
Grasshopper (Caelifera), Baekgangjam ( Bombysis Corpus ), white-spotted flower radish ( Protaetia brevitarsis seulensis ) larvae and twin- star cricket (Gryllus bimaculatus) contains an insect extract of at least one,
The insect extract is obtained at room temperature using an aqueous solution containing ascorbic acid as an extraction solvent, a food composition for improving cancer.

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