KR20190110510A - A composition for anti-cancer comprising the extract of herbal mixture, and uses thereof - Google Patents

Abstract

An embodiment of the present invention provides an anticancer composition containing a Curcuma longa extract, a Rhois Vernicifluae extract, and a Salvia miltiorrhiza extract as active components. According to the present invention, the anticancer composition not only effectively inhibits the growth of cancer, but also originates from natural herbal medicines, thereby not causing side effects on the human body.

Description

Anticancer composition comprising mixed herbal extracts and use thereof {A COMPOSITION FOR ANTI-CANCER COMPRISING THE EXTRACT OF HERBAL MIXTURE, AND USES THEREOF}

The present invention relates to an anticancer composition comprising a mixed herbal extract as an active ingredient.

Cancer is one of the incurable diseases that humanity has to solve, and huge capital is invested in the development to cure it all over the world.In Korea, it is the number one disease death cause and more than 100,000 people a year It is diagnosed, and about 60,000 or more die.

Carcinogens that cause cancer include smoking, ultraviolet rays, chemicals, foods, and other environmental factors. However, the causes of various cancers are difficult to develop, as well as the effects of treatments vary depending on the site of occurrence.

Despite the recent rapid development of medicine and various forms of cancer treatment through surgery, radiation therapy, and chemotherapy, many cancer patients still suffer from complete removal of cancer cells and prevention of metastasis.

The substances currently used as therapeutic agents are highly toxic and do not selectively remove only cancer cells. Therefore, there is an urgent need for development of low-toxic and effective anti-cancer drugs to prevent cancer after treatment. There is a situation.

As a result of continuing efforts to develop novel anticancer agents, the present inventors have found that mixed herbal extracts including turmeric, chilpi, and Salviae Radix can effectively inhibit the proliferation of cancer cells without affecting the growth of normal cells. By confirming that the present invention was completed.

 Anticancer natural products and their analogues, The Korean Society of Pharmacognosy, 17 (4), 286 291 (October 26, 1986)

An object of the present invention is to provide an anti-cancer composition excellent in biosafety and cancer inhibitory activity, including herbal extracts.

According to one aspect of the invention, there is provided an anticancer composition comprising turmeric extract, chilpi extract, and Salvia extract as an active ingredient.

In one embodiment, the composition may include 300 to 700 parts by weight of the turmeric extract, 30 to 70 parts by weight of the chilpi extract, and 5 to 15 parts by weight of the salvia extract.

In one embodiment, the turmeric extract and dansam extract may be an ethanol extract.

In one embodiment, the chilpi extract may be a hydrothermal extract.

In one embodiment, the cancer may be at least one selected from the group consisting of liver cancer, stomach cancer, colon cancer, lung cancer, breast cancer, rectal cancer and pancreatic cancer.

According to another aspect of the present invention, there is provided a pharmaceutical composition for the prevention or treatment of cancer comprising the composition.

According to another aspect of the present invention, a health functional food for preventing or improving cancer comprising the composition is provided.

According to another aspect of the present invention, there is provided an anticancer adjuvant composition comprising the composition.

According to the present invention, the anticancer composition not only effectively inhibits the growth of cancer, but also originates from natural herbal medicines and does not cause side effects on the human body.

It is to be understood that the effects of the present invention are not limited to the above effects, and include all effects deduced from the configuration of the invention described in the detailed description or claims of the present invention.

Figure 1 shows the injection of Sarcoma-180 cancer cells to induce celiac cancer and the survival date until 4 weeks (* p <0.05, ** p <0.01 versus the control group).
Figure 2 is a sarcoma-180 cancer cells were injected to induce solid cancer and tumor weight was measured after 4 weeks (* p <0.05, ** p <0.01 versus the control group).
Figure 3 is a LLC cancer cells were injected to induce solid cancer and measured the size of solid cancer for 3 weeks (** p <0.01, *** p <0.001 versus the control group).
4 shows tumor weight measured 3 weeks after induction of solid cancer by injection of LLC cancer cells (* p <0.05, *** p <0.001 versus the control group).
Figure 5 is a SNU-601 cell injected to induce solid cancer and measured the size of solid cancer for 6 weeks (* p <0.05, ** p <0.01, *** p <0.001 versus the control group).
Figure 6 is a SNU-601 cells injected to induce solid cancer and the tumor weight was measured after 6 weeks (* p <0.05, ** p <0.01, *** p <0.001 versus the control group).
7 is a visual observation of the tumor count of the lungs extracted 18 days after the injection of B16-F10 cells.
FIG. 8 shows inflammatory cytokines measured in the experimental group induced by celiac cancer by injecting Sarcoma-180 cancer cells (#p <0.05 versus normal group; * p <0.05 versus the control group).

The terminology used herein is to select general terms that are currently widely used as possible in consideration of the functions in the present invention, but may vary according to the intention or precedent of the person skilled in the art, the emergence of new technologies and the like. In addition, in certain cases, there is also a term arbitrarily selected by the applicant, in which case the meaning will be described in detail in the description of the invention. Therefore, the terms used in the present invention should be defined based on the meanings of the terms and the contents throughout the present invention, rather than the names of the simple terms.

Unless defined otherwise, all terms used herein, including technical or scientific terms, have the same meaning as commonly understood by one of ordinary skill in the art. Terms such as those defined in the commonly used dictionaries should be construed as having meanings consistent with the meanings in the context of the related art, and are not construed in ideal or excessively formal meanings unless expressly defined in this application. Do not.

The numerical range includes the numerical values defined in the range. All maximum numerical limits given throughout this specification include all lower numerical limits as if the lower numerical limits were clearly written. All minimum numerical limits given throughout this specification include all higher numerical limitations as if the higher numerical limit were clearly written. All numerical limitations given throughout this specification will include all better numerical ranges within the broader numerical range, as the narrower numerical limitations are clearly written.

When a part is said to "include" a certain component, this means that it may further include other components, without excluding other components, unless specifically stated otherwise.

Hereinafter, with reference to the accompanying drawings will be described the present invention. As those skilled in the art would realize, the described embodiments may be modified in various different ways, all without departing from the spirit or scope of the present invention.

According to one aspect of the invention, there is provided an anticancer composition comprising turmeric extract, chilpi extract, and Salvia extract as an active ingredient.

Said “cancer”, “tumor” or “malignant” generally refers to or describes the physiological state of a mammal that is characterized by unregulated cell growth. Examples of cancer include, but are not limited to, carcinoma, lymphoma, leukemia, blastoma, and sarcoma, and may be selected, for example, from the group consisting of liver cancer, gastric cancer, colon cancer, lung cancer, breast cancer, rectal cancer, and pancreatic cancer.

The "comprising as an active ingredient" may mean that it contains an effective amount that can exhibit an anticancer effect, for example, induces the death of cancer cells or inhibits the proliferation of cancer cells.

The "extract" is to isolate the active ingredient contained in the extract raw material by contacting the solvent and the extract raw material under specific conditions, the extract may include a variety of active ingredients contained in the herbal medicine.

The “Ulgeum (鬱金, Curcuma aromatica Salisb. )” Is a perennial ripening herbaceous plant, which is a turmeric root of turmeric, turmeric, and azu.

The turmeric affects lipid metabolism (animal experiment) according to the antibiotic (medicinal) metabolism, can reduce the formation of endometrial masses of the main and coronary arteries, deposition of lipids, and contained essential oils secrete and excrete bile It is known to be used for cholelithiasis because it has an effect of promoting gall bladder contraction and has an biliary function and dissolves filamentous stones. In addition, according to the Chinese herbal medicine dictionary, the taste is spicy and bitter, and the nature is limited and nontoxic, and it promotes the circulation of the qi, loosening the loose, bleeding blood and removing the blood. The turmeric is known to treat chest pain, ecchymosis, fever newlyweds, hemorrhage, nasal bleeding, hematuria, hemolimb, menstruation of men and jaundice.

The bark of the lacquer belongs to the " Rhus verniciflua Stokes " family Anacardiaceae . The lacquer tree is a deciduous broad-leaved arboreous tree that grows a lot in Northeast Asia such as China and Japan. From Chilpi, its bark, phenolic compounds such as chalcone and flavonoids have been isolated and reported to date. Recently, anti-cancer, anti-inflammatory, antioxidant, hepatocellular protective activity, anti-apoptosis, and immunomodulatory activity of chilpi extract or phenolic compounds isolated therefrom have been reported.

The “danshen ( Salvia miltiorrhiza Bunge )” is a perennial plant belonging to the Lamiaceae ( Labiatae ), and has the effect of activating live blood, and hematopoietic swelling in the Chinese medicine. It is used as a medicine for menstrual pain, postpartum abdominal pain and hemolytic heart pain, bruises, insomnia and skin rash.

The main chemical components of the Salvia Militiorrhiza are diterpene compounds including tanshinone I, IIA, IIB, salvianic acid A, protocatechuic aldehyde, and salvia. Phenolic compounds, including salvianolicacid B, etc., in addition to ical compounds such as baicaline, β-sitosterol, ursolic acid, vitamin E and tannin ( tannin) and the like, and the physiological activity of the salviae, such as antibacterial, antioxidant, anticancer, antimutagenic activity has been known.

For example, the herbal extracts are washed with water, dried, and finely pulverized with each herbal medicine, and a reflux circulation extraction, pressure extraction, ultrasonic extraction, etc., is performed for about 2 to 5 hours with a solvent having a volume of 5 to 20 times the weight of each raw material. It can be prepared by extraction and filtration by conventional methods. In addition, the extract may be obtained in a powder state by an additional process such as distillation under reduced pressure or freeze drying.

On the other hand, the extract may include an extract that has undergone a conventional purification process. For example, the extract may be subjected to various purification methods additionally performed, such as separation using an ultrafiltration membrane having a constant molecular weight cut-off value, separation by various chromatography (manufactured for separation according to size, charge, hydrophobicity, or affinity). It may comprise a fraction obtained through.

The kind of solvent is not specifically limited in the said extraction process, Various solvent can be used as needed. The extraction may be used by the conventional methods in the art, such as chilling, warming, heating using the solvent, preferably the turmeric extract and dansam extract may be an ethanol extract, the chilpi extract may be a hydrothermal extract have.

Since the active ingredient included in the raw material may be different in the extraction ratio according to the polarity of the solvent, the characteristics of the extraction solvent may be varied in order to maximize the content of the active ingredient exhibiting anticancer activity contained in the raw material.

Based on thousands of clinical treatments, the inventors have applied optimized solvents to achieve excellent anticancer effects. In particular, water and ethanol are excellent in selectivity in the extraction of the physiologically active substance contained in the herbal medicine, so the optimum anticancer effect can be realized by the solvent.

Since water and ethanol are different in polarity, the active ingredient extracted according to each polarity may be different, so that the concentration of the ethanol can be appropriately controlled so that an optimal anticancer effect can be realized.

However, when the concentration of ethanol is more than 90% (v / v) may not be achieved a proper yield, if the concentration is less than 60% (v / v) may not be extracted enough active ingredients exhibiting an anticancer effect. have.

The composition may include 300 to 700 parts by weight of the turmeric extract, 30 to 70 parts by weight of the chilpi extract, and 5 to 15 parts by weight of the salvia extract.

When the mixed extract has a large variation in the mixing ratio of the raw materials, the anticancer effect derived from the active ingredient of each extract may not be properly implemented, and the mixing ratio may be properly controlled in consideration of the working environment and the quality of the final product. .

The inventors have derived the optimum mixing ratio of the extract based on the clinical results, the anticancer activity of the composition can be maximized within the range.

In addition, the turmeric extract, chilpi extract, and Dansam extract may be contained 1.0 to 10.0% by weight based on the total weight of the composition.

The extract may be difficult to obtain the expected effect if the content is less than 0.1% by weight, the excess of 10.0% by weight may be excessively economical or the original quality of the applied product may not be implemented.

According to another aspect of the present invention, there is provided a pharmaceutical composition for the prevention or treatment of cancer comprising the composition.

The term "prevention" means any action that reduces the frequency or extent of pathological phenomena. Prevention can be complete or partial. In this case, compared to the case where the composition is not used, it may mean that the growth of cancer cells in an individual is suppressed or the cancer cells are killed.

The term "treatment" refers to all actions that are clinically involved in altering the natural process of the subject or cell to be treated, and may be performed during or to prevent a clinical pathology. The desired therapeutic effect may prevent the occurrence or recurrence of the disease, alleviate the symptoms, reduce all direct or indirect pathological consequences of the disease, prevent metastasis, slow the progression of the disease, or Mitigating or temporarily alleviating the condition or improving the prognosis.

The composition may be administered in the form of oral delivery, parenteral delivery. The composition can be administered systemically or topically, and the administration can include oral and parenteral administration. The composition may be formulated with a suitable amount of pharmaceutically acceptable vehicle or carrier to provide a suitable dosage form.

The composition may further comprise a carrier, excipient and diluent used in the manufacture of the pharmaceutical composition. The carrier, excipient and diluent may include lactose, dextrose, sucrose, sorbitol, mannitol, xylitol, erythritol, maltitol, starch, acacia rubber, alginate, gelatin, calcium phosphate, calcium silicate, cellulose, methyl cellulose, microcrystalline Cellulose, polyvinyl pyrrolidone, water, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate or mineral oil, but are not limited thereto.

In addition, the composition may be formulated in the form of powders, granules, tablets, capsules, suspensions, emulsions, syrups, aerosols and the like, oral formulations, suppositories, and sterile injectable solutions.

Solid preparations for oral administration may be used in tablets, pills, powders, granules, capsules, and the like, wherein the solid preparations include at least one excipient such as starch, calcium carbonate, sucrose, lactose in the compound and fractions thereof. Or gelatin can be mixed and prepared. In addition to the above excipients, lubricants such as magnesium styrate and talc may be used.

As a liquid preparation for oral administration, suspending agents, solvents, emulsions, syrups, and the like may be used, and various excipients such as wetting agents, sweeteners, fragrances, preservatives, and the like may be used in addition to water and liquid paraffin, which are simple diluents.

For parenteral administration, sterile aqueous solutions, non-aqueous solvents, suspensions, emulsions, lyophilized formulations, suppositories can be used. The non-aqueous solvent and suspension may be propylene glycol, polyethylene glycol, vegetable oils such as olive oil, injectable esters such as ethyl oleate. As the base of the suppository, witepsol, macrogol, tween 61, cacao butter, laurin butter, and glycerogelatin may be used.

The composition may be administered in a pharmaceutically effective amount sufficient to treat the disease at a reasonable benefit / risk ratio applicable to the medical treatment, and the effective dose level may be of the type and severity of the subject, age, sex, type of disease, drug Activity, sensitivity to drug, time of administration, route of administration and rate of release, duration of treatment, factors including concurrently used drugs, and other factors well known in the medical arts.

The composition may be administered as a separate therapeutic agent or in combination with other therapeutic agents such as anticancer agents, may be administered sequentially or simultaneously with conventional therapeutic agents, and may be single or multiple doses.

The anticancer agent is cisplatin, oxaliplatin, carboplatin, carcarplatin, procarbazine, mechlorethamine, cyclophosphamide, ifosfamide, Melphalan, chlorambucil, bisulfan, nitrosourea, diactinomycin, daunorubicin, doxorubicin, bleomycin , Plicomycin, mitomycin, etoposide, tamoxifen, taxol, transplatinum, 5-fluorouracil , Vincristin, vinblastin, methotrexate, but are not limited thereto.

Taking all of the above factors into consideration, it is important to administer an amount that can obtain the maximum effect in a minimum amount without side effects, and can be easily determined by those skilled in the art.

The preferred dosage of the composition may vary depending on the condition and weight of the patient, the severity of the disease, the form of the drug, the route of administration, and the duration of time, and a suitable total daily dose may be determined by the practitioner within the correct medical judgment. . Generally, an amount of 0.001 to 1000 mg / kg, preferably 0.05 to 200 mg / kg, more preferably 0.1 to 100 mg / kg, may be administered once to several times daily.

The composition is not particularly limited as long as it is an individual for the purpose of preventing or treating cancer, and any individual may be applied. For example, it can be applied to any individual such as non-human animals such as monkeys, dogs, cats, rabbits, marmots, rats, mice, cows, sheep, pigs, goats, and humans, and the like, and the mode of administration is limited if it is a conventional method in the art. Includes without.

According to another aspect of the present invention, a health functional food for preventing or improving cancer comprising the composition is provided.

The health functional food means a food manufactured and processed using raw materials or ingredients having functional properties useful for the human body according to the law on health functional foods, and the functional properties control nutrients or physiologically on the structure and function of the human body. It may mean ingestion for the purpose of obtaining a useful effect for health use such as action.

The health functional food may include a conventional food additive, and unless otherwise specified, the food additive may comply with the standards and standards for the item in accordance with the General Regulations and General Test Act of the Food Additives Act approved by the Ministry of Food and Drug Safety. Compliance can be determined by

The items described in the food additive orbital are, for example, chemical compounds such as ketones, glycine, potassium citrate, nicotinic acid, and cinnamon acid, natural additives such as color pigments, licorice extract, crystalline cellulose, high pigment, guar gum, sodium L-glutamate preparations, and noodles. Mixed preparations, such as an added alkali, a preservative, and a tar pigment, are mentioned.

The health functional food may be used in a variety of foods and drinks for the prevention or improvement of alopecia, for example, it can be used in various foods, beverages, gum, tea, vitamin complexes, health functional supplements, food additives and the like.

The health functional food may be prepared and processed in any one formulation selected from the group consisting of tablets, granules, powders, capsules, liquid solutions and pills for the purpose of preventing or improving cancer.

Specifically, the health functional food in the form of tablets is a mixture of the compound, excipients, binders, disintegrants and other additives in a conventional manner, and then compression-molded with a lubricant, or directly compression-molded the mixture Can be prepared. In addition, the health functional food in the form of tablets may contain a mating agent and the like, if necessary, may be coated with a suitable coating agent.

The hard capsules of the health functional food in the form of capsules may be prepared by filling a conventional hard capsule with a mixture of additives such as the compound and excipients or granules or peeled granules thereof, and the soft capsules may contain the compounds and excipients. The mixture with additives, such as these, can be prepared by filling in capsule bases, such as gelatin. The soft capsule agent may contain a plasticizer such as glycerin or sorbitol, a colorant, a preservative, and the like, as necessary.

The dietary supplement in the form of a cyclic form may be prepared by molding a mixture of the compound, excipient, binder, disintegrating agent, etc. in a suitable manner, and may be coated with sucrose or other suitable coating agent, or starch, talc, or a suitable substance. You can also give a favor.

The granular health functional food may be prepared by granulating a mixture of the compound, excipient, binder, disintegrating agent and the like in an appropriate manner, and may contain a flavoring agent, a copper, and the like as necessary.

In addition, the definitions of the excipients, binders, disintegrants, glidants, copulation agents, flavoring agents and the like are described in documents known in the art and may include those having the same or similar functions.

Those skilled in the art to which the present invention pertains will be able to apply by changing the type, introduction ratio, etc. of each configuration based on the description of the present invention, if the equivalent technical effect is implemented in spite of the above modification, the present invention It will be included in the technical idea of the.

According to another aspect of the present invention, there is provided an anticancer adjuvant composition comprising the composition.

The "anticancer adjuvant" means an agent that can improve, enhance or enhance the anticancer effect of the anticancer agent. For example, when the anticancer adjuvant is used in combination with an anticancer agent, the anticancer effect of the anticancer agent may be improved, improved or increased.

As another example, a concentration-dependent anticancer agent may be used as an anticancer adjuvant that can improve, enhance or enhance the anticancer effect of the anticancer agent when used in combination with an anticancer agent at a level that does not exhibit anticancer activity by itself. . The anticancer adjuvant may be used as an anticancer agent or an anticancer adjuvant according to the treatment concentration, and may be used as an anticancer adjuvant in the treatment concentration range which does not exhibit anticancer activity by itself.

Through the following examples, the present invention will be described in more detail, but the present invention is not limited by the following examples.

Example 1

500 g turmeric was refluxed with 80% (v / v) ethanol for 2 hours and concentrated to give 46.0 g as a lyophilized product (CLR, yield 9.20%).

700 g of chile was digitized at 180 ° C. for 1 hour, and then extracted and concentrated for 2 hours with distilled water (DW) to give 24.0 g of lyophilized product (RRVS, yield 3.41%).

200 g of Salvia ginseng was refluxed with 80% (v / v) ethanol for 2 hours and concentrated to give 45.0 g as a lyophilized product (SMB, yield 22.5%).

Cultivated red ginseng extract mixed with CLR, RRVS and SMB in a weight ratio of 50: 5: 1 was named CHD.

Example 2

Prepared in the same manner as in Example 1, each herbal medicine was extracted with distilled water (DW) hot water.

Example 3

Prepared in the same manner as in Example 1, each herbal extract was refluxed with 80% (v / v) ethanol.

Comparative example

The following herbal extracts were set as comparative examples. The preparation method of the extract was the same as in Examples 1-3.

[Contents (weight ratio)] division Comparative example One 2 3 4 5 6 7 Turmeric ethanol extract 150 - - 75 - 75 50 Chilpi hot water extract - 150 - 75 75 - 50 Sweet Ginseng Ethanol Extract - - 150 - 75 75 50

Experiment method

Cancer cell line culture

Colon cancer cells LoVo, breast cancer cells MCF-7, liver cancer cells HepG2, gastric cancer cells AGS, non-small cell lung cancer cells A549, small cell lung cancer cells NCI-H69, leukemia cells HL-60, celiac cancer cells Sarcoma 180 was sold by the Korean Cell Line Bank.

HepG2 in MEM medium (gibco, USA) with 10% FBS (Gibco, USA) and 1% penicillin-streptomycin (Gibco, USA), other cells were RPMI with 10% FBS and 1% penicillin-streptomycin Cultured in a medium (Gibco, USA) incubator at 37 ℃, 5% CO ₂ concentration.

Lung cancer cells, LL / 2, were purchased from the American Type Culture Collection (ATCC, USA) and incubator at 37 ° C and 5% CO ₂ concentration in DMEM medium (ATCC, USA) with 10% FBS and 1% penicillin-streptomycin. Incubated in the.

Toxicity measurement on cancer cell lines

Cytotoxicity experiments were measured using the MTT assay method to measure the degree of reduction of 3- (4,5-dimethylthiazol-2yl) -2,5-diphenyl tetrazolium bromide (MTT).

Each cell line was seeded at 2 x 10 ⁴ cells / well in a 96 well plate and incubated for 24 hours, the sample was treated with a concentration of 0.1 to 1 mg / mL.

After 48 hours of incubation, 25 μL / mL of 1 mg / mL MTT solution (Sigma, USA) was added to each well and incubated for 4 hours in an incubator at 37 ° C., 5% CO ₂ .

After removing the medium, 50 μL / mL of DMSO (dimethyl sulfoxide) was added to dissolve the insoluble formazan crystals, and the absorbance was measured at 540 nm with an ELISA reader.

Suspension cells NCI-H69 and HL-60 were treated with the same method as above, and the absorbance was measured at 540 nm using the EZ-cytox kit (Daeillab Service Co., LTD, Korea).

Laboratory animals

The experimental animals were 6-week-old male C57BL / 6, Balb / c mouse, Balb / c nude mouse (Central laboratory animals, Seoul) or ICR male mouse, SD rat (Young Bio, Seongnam), and the temperature was 22 ± 2 ℃. The diet and drinking water were allowed to be consumed freely in a controlled breeding room with a humidity of 60 ± 10% and a 12 hour light-dark cycle.

Management and maintenance of laboratory animals and all experiments were conducted with the approval of Kyung Hee Medical Center's Laboratory Animal Ethics Committee.

Celiac Cancer Model Induced with Sarcoma-180

To induce celiac cancer, 0.1 mL (1.0 × 10 ⁶ cells / mouse) of sarcoma-180 cell suspension was injected into the abdominal cavity of ICR mice.

After 24 hours after the intraperitoneal cancer cell transplantation, physiological saline or experimental drug was administered orally for 4 consecutive weeks.

As a positive control, cyclophosphamide was intraperitoneally administered at a concentration of 10 mg / kg. Survival was observed for 4 weeks after transplantation of cancer cells in the abdominal cavity, and the life extension percentage was calculated according to the following formula.

Life extension percentage (%) = average lifespan (days) of experimental group-average lifespan (days) of control group / average lifespan (days) of control group x 100

Solid Cancer Model Induced by Sarcoma-180

To induce solid cancer, 0.3 mL (2.0 × 10 ⁷ cells / mouse) of sarcoma-180 cell suspension was injected into the groin of ICR mice.

After 24 hours after transplanting the cancer cells, physiological saline or experimental drug was administered orally for 4 consecutive weeks.

As a positive control, cyclophosphamide was intraperitoneally administered at a concentration of 20 mg / kg. Four weeks after the implantation of cancer cells in the inguinal part, the solid tumors produced were isolated and weighed. Tumor growth inhibition was calculated as follows.

Tumor growth retardation (%) = solid cancer weight of control group (g)-solid cancer weight of experimental group (g) / solid cancer weight of control group (g) × 100

LLC-induced solid cancer model

In order to induce solid cancer using LLC (Lewis lung cancer cell), 0.1 mL (1.5 × 10 ⁶ cells / mouse) of LLC was injected subcutaneously into the back of C57BL / 6 mice.

Twenty-four hours after cancer cell transplantation, saline or experimental drug was orally administered for 3 consecutive weeks. Cyclophosphamide was intraperitoneally administered at a concentration of 20 mg / kg as a positive control.

Tumor size was measured twice a week for 3 weeks, and solid tumors were removed and weighed at the end of the experiment. Tumor growth hypothesis was calculated as follows.

Tumor growth retardation (%) = solid cancer weight of control group (g)-solid cancer weight of experimental group (g) / solid cancer weight of control group (g) × 100

Gastric cancer model induced by SNU-601

In order to induce solid cancer using SNU-601 cells, 0.1 mL (1.0 × 10 ⁷ cells / mouse) of SNU-601 cells were subcutaneously injected into the back of Balb / c nude mice.

Twenty four hours after cancer cell transplantation, saline or experimental drug was administered orally continuously for 6 weeks. The positive control group was intraperitoneally administered with cyclophosphamide at a concentration of 20 mg / kg. Tumor size was measured twice a week for 6 weeks, and solid tumors were extracted and weighed at the end of the experiment. Tumor growth hypothesis was calculated as follows.

Tumor growth retardation (%) = solid cancer weight of control group (g)-solid cancer weight of experimental group (g) / solid cancer weight of control group (g) × 100

Lung cancer metastasis model induced by melanoma

To induce metastatic cancer using Melanoma cells, 0.1 ml (6.0 × 10 ⁵ cells / mouse) of B16-F10 cells were administered to the tail vein of C57BL / 6 mice.

Cancer cells were administered and physiological saline or experimental drug was orally administered for 18 consecutive days after 24 hours had elapsed. As a positive control, cyclophosphamide was intraperitoneally administered at a concentration of 20 mg / kg. After 18 days, the lungs were removed by autopsy, and the number of tumors was visually observed and compared.

Azoxymethane-induced Colorectal Cancer Model

To induce colorectal cancer in balb / c mice, azoxymethane was intraperitoneally injected at a dose of 25 mg / kg.

Two weeks after the administration of Azoxymethane, 2% dextran sulfate sodium (DSS) was negatively administered for one week, followed by a two-week rest period, followed by a DSS-negative administration for one week.

The test drug was orally administered two weeks after the azoxymethane administration, and the animals were necropsied after a total of eight weeks.

The large intestine of the mouse was colonized from the anus to the ilocecal junction, and the length from the cecum to the anus was measured by weight and weighed. Tumor counts were compared by visual observation.

Diethylnitrosamine-induced Liver Cancer Model

To induce liver cancer in S.D rats, 50 mg / kg of diethylnitrosamine (MO, U.S.A.) was dissolved in 2 ml of 0.9% saline and injected 12 times per week. The test drug was administered after diethylnitrosamine intraperitoneal injection for a total of 8 weeks.

After the end of the experiment, livers were collected and visually counted and compared with the number of tumors. Blood was collected to measure factors such as GOT, GPT, and ALP.

Measurement of Immune Factors in Sarcoma-180-Induced Solid Cancer Models

To induce solid cancer, 0.3 mL (2.0 × 10 ⁷ cells / mouse) of sarcoma-180 cell suspension was injected into the groin of ICR mice.

After cancer cell transplantation, physiological saline or experimental drug was orally administered for 4 consecutive weeks after 24 hours.

Four weeks after transplanting the cancer cells in the inguinal part, blood was collected from the heart, centrifuged at 3000 rpm for 10 minutes, and serum was used for this experiment. Serum assays were determined using IFN-γ, IL-2 and IL-4 ELISA kits (Thermo Fisher Scientific, U.S.A).

Statistical verification

The results obtained in this experiment were expressed as mean ± standard error (S.E.M), and One-way ANOVA test was conducted to test the significance.

For post hoc test, Dunnett's test was used for comparison. Statistical significance level was p <0.05.

Experiment result

Effect on Cancer Cell Toxicity

To examine the cancer cell proliferation inhibitory effect of Examples and Comparative Examples, colon cancer cells LoVo, breast cancer cells MCF-7, liver cancer cells HepG2, gastric cancer cells AGS, non-small cell lung cancer cells A549, small cell lung cancer cells NCI- MTT assay was performed on H69, HL-60, a leukemia cell, Sarcoma 180, a celiac cancer cell, and LL / 2, a lung cancer cell.

(IC50, mg / mL) division Example Comparative example One 2 3 One 2 3 4 5 6 7 LoVo <0.10 <0.10 <0.10 0.11 0.16 0.14 0.16 0.14 0.21 <0.10 MCF-7 0.25 0.29 0.30 0.34 0.37 0.35 0.41 0.36 0.33 0.30 HepG2 0.50 0.43 0.55 0.55 0.57 0.61 0.59 0.57 0.63 0.59 AGS 0.16 0.19 0.26 0.27 0.27 0.27 0.27 0.27 0.27 0.28 A549 0.32 0.45 0.39 0.44 0.48 0.51 0.54 0.49 0.41 0.49 NCI-H69 <0.10 <0.10 <0.10 <0.10 0.13 0.14 0.13 0.13 0.11 <0.10 HL-60 <0.10 <0.10 <0.10 <0.10 0.11 0.13 <0.10 <0.10 0.12 <0.10 Sarcoma 180 0.18 0.27 0.34 0.41 0.41 0.41 0.41 0.41 0.41 0.26 LL / 2 <0.10 <0.10 <0.10 <0.10 0.11 0.13 0.14 0.11 0.12 <0.10

Comparative Example 1 (CLR) at a concentration of 1 mg / mL or less had a cancer cell proliferation inhibitory effect of more than 50% for all the cancer cell lines above, Example 1 (CHD) is especially NCI-H69, HL-60 and LL / For 2 cells, the IC50 value was 0.10 mg / mL or less, showing a strong inhibitory effect.

Example 1 (CHD) had a cancer cell proliferation inhibitory effect of at least 50% for all cancer cell lines above at a concentration of 1 mg / mL, in particular IC50 values for LoVo, NCI-H69, HL-60 and LL / 2 cells Strong inhibitory activity was shown at 0.10 mg / mL or less.

Inhibitory Effect on Sarcoma-180 Induced Celiac Cancer

The anticancer effect on Sarcoma-180 cancer cells of Examples and Comparative Examples was measured. Sarcoma-180 cancer cells were injected into the abdominal cavity of experimental animals to induce celiac cancer and survival was measured until 4 weeks later (FIG. 1).

The control group administered with saline died on an average of 17.6 days, but the control group of Comparative Example 1 (CLR) died on an average of 19.3 days, and the control group of Example 1 (CHD) died on an average of 20.2 days. It was increased to 10.1 and 15.1%, respectively.

Sarcoma-180 Induced Solid Cancer Growth Inhibition

In order to measure the anticancer effect on Sarcoma-180 cancer cells, Sarcoma-180 cancer cells were injected into the mouse groin to induce solid cancer and tumor weight was measured 4 weeks later (FIG. 2).

In the solid cancer model induced by Sarcoma-180, the solid cancer weight of the control group averaged 8.0 g, but the solid cancer weight of the Comparative Example 1 (CLR) group was 5.5 g on average and the solid cancer weight of the Example 1 (CHD) group was 4.5 g on the average. Compared with that, the weight of solid cancer was significantly reduced and tumor growth inhibition was 31.6% and 44.5%, respectively.

Cyclophosphamide, a positive control group, averaged 2.9 g, which showed about 63.8% of tumor growth inhibition effect.

LLC-induced solid cancer growth inhibition effect

In order to measure the effect of inhibiting the growth of solid cancers of Examples and Comparative Examples, LLC cancer cells were injected into the embryos of experimental animals to induce solid cancers, and the size of solid cancers was measured for three weeks (FIG. 3). 4).

The solid cancer size of the control group administered with saline was 6.9 cm ³ on average, but the solid cancer size of the administration group of 1 g / kg of Comparative Example 1 (CLR) was 5.6 cm ³ , indicating that the size of solid cancer decreased compared to the control group.

Example 1 (CHD) solid cancer size of 1g / kg administration group was confirmed to be significantly reduced to 4.1cm ³ and showed a tumor growth inhibitory effect of about 41.3%.

The average size of solid cancer in the positive control cyclophosphamide group was 2.7 cm ^{3, which} was 61.5% lower than the control group.

The average weight of solid cancer in the control group administered with saline was 5.1g, but the weight of solid cancer in the 1g / kg administration group of Comparative Example 1 (CLR) averaged 4.2g.

Example 1 (CHD) 1g / kg administration of the average solid cancer weight of 3.5g compared to the control group significantly reduced the weight of the solid cancer showed about 30.5% tumor growth inhibition effect. The mean weight of solid cancer in the group treated with cyclophosphamide, a positive control group, was 2.1 g, which was about 58.5% less than that of the control group.

SNU-601 Induced Gastric Cancer Growth Inhibitory Effect

In order to measure the anticancer effect on SNU-601 cancer cells of the Examples and Comparative Examples, SNU-601 cells were injected into the embryos of experimental animals to induce solid cancer and measure the size of solid cancer for 6 weeks (FIG. 5). Was measured (FIG. 6).

Solid tumor size in the control group administered with physiological saline average yieoteuna 2.6cm ^3, Comparative Example 1 (CLR) solid tumor size of 1g / kg treated group on average 1.8 cm ^3, Example 1 (CHD) solid tumor size of 1g / kg treated group was 1.3 the size of the solid cancer was reduced by 28.9 each, 48.6% compared to the control group ³ cm.

In the positive control group, cyclophosphamide, the average of 0.2cm ³ showed tumor suppression effect of 90.7%.

The average weight of solid cancer in the control group administered with saline was 2.2 g, but the average weight of solid cancer in the 1 g / kg administration group of Comparative Example 1 (CLR) was 1.1 g, and the average weight of the solid cancer in the 1 g / kg administration group of Example 1 (CHD) was 0.7 g. The weight of solid cancers was reduced by 52.4 and 70.7%, respectively. Cyclophosphamide, a positive control group, averaged 0.1 g and tumor growth inhibition effect was about 95.4%.

Inhibitory effect of melanoma-induced lung cancer metastasis

B16-F10 cells were injected into the tail vein of the experimental animals in order to measure the cancer metastasis inhibitory effect of the Examples and Comparative Examples, 18 days later, the lungs were removed and the number of tumors was visually observed (FIG. 7).

The number of tumors in the control group administered with saline was 92.2, but the number of tumors in the Comparative Example 1 (CLR) group was 74.2 and the number of tumors in the Example 1 (CHD) group was 64.2, compared to the control group. , 30.4%. In the control group treated with cyclophosphamide, 49.2 patients had an average of 46.7% of antitumor effect.

Azoxymethane-induced Colon Cancer Growth Inhibition

In order to measure the anticancer effect on azoxymethane-induced colorectal cancer of Examples and Comparative Examples, azoxymethane was injected into the abdominal cavity of experimental animals to induce colorectal cancer, and the length, weight and number of tumors of the colon were visually measured after 8 weeks. Table 3).

In comparison with the number of tumors in the colon, the average number of tumors was 7.1 in the control group, and a significant decrease was found in the comparative group 1 (CLR) administration group at an average of 6.0 and the average group in the administration of Example 1 (CHD) group at 5.4.

An average of 4.9 were observed in the positive control fluorouracil group. When the weight and length of the colon were measured, there was no difference in weight, but the length of the colon of the control group was shortened and significantly improved by administration of Comparative Example 1 (CLR) and Example 1 (CHD).

division Weight (g) Colon weight (g) Colon length (cm) Tumor count Normal 25.0 ± 0.7 0.43 ± 0.006 8.7 ± 0.16 - Control 24.2 ± 0.6 0.44 ± 0.017 7.1 ± 0.30 ^## 7.4 ± 0.6 Example 1 (CHD) 24.9 ± 0.4 0.42 ± 0.016 8.2 ± 0.31 ^* 5.4 ± 0.7 ^* Example 2 24.9 ± 0.4 0.41 ± 0.016 8.1 ± 0.31 ^* 5.6 ± 0.7 ^* Example 3 24.9 ± 0.4 0.42 ± 0.016 8.2 ± 0.31 ^* 5.5 ± 0.7 ^* Comparative Example 1 (CLR) 24.5 ± 0.4 0.41 ± 0.018 8.0 ± 0.24 ^* 6.0 ± 0.4 ^* Comparative Example 2 24.4 ± 0.4 0.41 ± 0.018 7.9 ± 0.24 ^* 6.1 ± 0.4 ^* Comparative Example 3 24.3 ± 0.4 0.41 ± 0.018 7.8 ± 0.24 ^* 6.1 ± 0.4 ^* Comparative Example 4 24.4 ± 0.4 0.40 ± 0.018 7.9 ± 0.24 ^* 6.2 ± 0.4 ^* Comparative Example 5 24.5 ± 0.4 0.39 ± 0.018 7.7 ± 0.24 ^* 6.1 ± 0.4 ^* Comparative Example 6 24.6 ± 0.4 0.40 ± 0.017 7.9 ± 0.24 ^* 6.1 ± 0.4 ^* Comparative Example 7 24.6 ± 0.4 0.41 ± 0.016 8.1 ± 0.31 ^* 5.8 ± 0.4 ^* Fluorouracil 24.3 ± 0.3 0.43 ± 0.022 8.3 ± 0.20 ^* 4.9 ± 0.7 ^*

(## p <0.01 versus normal group; * p <0.05 versus control group)

Inhibitory Effect of Diethylnitrosamine-induced Growth on Liver Cancer

The anticancer effect of diethylnitrosamine-induced liver cancer of Examples and Comparative Examples was measured (Table 4). The average number of tumors in the control group was 10.3, and the number of tumors in the Comparative Example 1 (CLR) -administered group tended to decrease to 7.2, and the number of tumors in the Example 1 (CHD) -administered group was 4.8.

GOT, GPT, and ALP all significantly increased in the control group, and administration of Comparative Example 1 (CLR) and Example 1 (CHD) significantly decreased it.

division GOT (U / L) GPT (U / L) ALP (U / L) Tumor count Normal 140 ± 34.5 61 ± 3.1 490 ± 21.8 - Control 315 ± 20.8 ^# 107 ± 14.3 ^# 799 ± 21.8 ^# 10.3 ± 2.2 ^# Example 1 (CHD) 207 ± 35.1 ^* 67 ± 4.0 ^* 499 ± 23.4 ^* 4.8 ± 0.8 ^* Example 2 208 ± 35.1 ^* 69 ± 4.0 ^* 507 ± 23.4 ^* 5.2 ± 0.8 ^* Example 3 209 ± 35.1 ^* 70 ± 4.0 ^* 502 ± 23.4 ^* 5.3 ± 0.8 ^* Comparative Example 1 (CLR) 221 ± 19.6 ^* 82 ± 3.8 ^* 542 ± 36.9 ^* 7.2 ± 1.1 Comparative Example 2 223 ± 19.6 ^* 81 ± 3.8 ^* 545 ± 36.9 ^* 7.4 ± 1.1 Comparative Example 3 221 ± 19.6 ^* 83 ± 3.8 ^* 544 ± 36.9 ^* 7.3 ± 1.1 Comparative Example 4 222 ± 19.6 ^* 79 ± 3.8 ^* 541 ± 36.9 ^* 7.5 ± 1.1 Comparative Example 5 221 ± 19.6 ^* 84 ± 3.8 ^* 543 ± 36.9 ^* 7.4 ± 1.1 Comparative Example 6 218 ± 19.6 ^* 85 ± 3.8 ^* 546 ± 36.9 ^* 7.1 ± 1.1 Comparative Example 7 216 ± 19.6 ^* 72 ± 3.8 ^* 510 ± 36.9 ^* 6.3 ± 1.0 Cyclophosphamide 196 ± 19.3 ^* 61 ± 6.3 ^* 457 ± 25.8 ^* 2.8 ± 1.5 ^*

(#p <0.05 versus normal group; * p <0.05 versus control group)

Measure the effect on immunity

Representative cytokines, IFN-γ, IL-2 and IL-4, were measured to evaluate the effect of the samples of Examples and Comparative Examples on immunity (FIG. 8).

Sarcoma-180-induced solid cancer significantly reduced IFN-γ levels in the control group and this phenomenon was ameliorated by the administration of Comparative Example 1 (CLR).

In the Example 1 (CHD) administration group, the IFN-γ level was significantly increased compared to the control group, and the improvement effect was 20.2%.

As a result of IL-2 measurement, IL-2 levels were significantly decreased in the control group, but IL-2 levels were significantly increased in the Example 1 (CHD) -administered group, and the improvement effect was about 34.2%.

In addition, IL-4 levels were also significantly reduced in the control group and significantly improved by administration of Example 1 (CHD) (about 43.0%).

The foregoing description of the present invention is intended for illustration, and it will be understood by those skilled in the art that the present invention may be easily modified in other specific forms without changing the technical spirit or essential features of the present invention. will be. Therefore, it should be understood that the embodiments described above are exemplary in all respects and not restrictive. For example, each component described as a single type may be implemented in a distributed manner, and similarly, components described as distributed may be implemented in a combined form.

The scope of the invention is indicated by the following claims, and it should be construed that all changes or modifications derived from the meaning and scope of the claims and their equivalents are included in the scope of the invention.

Claims (7)

Which contains turmeric extract, chilpi extract, and salvia extract,
Anticancer composition of at least one cancer selected from the group consisting of liver cancer, stomach cancer, colon cancer, lung cancer, breast cancer, rectal cancer, pancreatic cancer and celiac cancer caused by sarcoma. The method of claim 1,
300 to 700 parts by weight of the turmeric extract, 30 to 70 parts by weight of the chilpi extract, and 5 to 15 parts by weight of the Salvia extract, anticancer composition. The method of claim 1,
The turmeric extract and dansam extract is an ethanol extract, the chilpi extract is a hot water extract, anticancer composition. The method of claim 1,
The anticancer composition is to inhibit the metastasis of one or more cancers selected from the group consisting of liver cancer, stomach cancer, colon cancer, lung cancer, breast cancer, rectal cancer, pancreatic cancer and celiac cancer caused by sarcoma. A pharmaceutical composition for preventing or treating cancer, comprising the composition of any one of claims 1 to 4. Claims 1 to 4, comprising the composition of any one of the health functional food for the prevention or improvement of cancer. Anticancer adjuvant composition containing the composition of any one of Claims 1-4.

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