KR100749115B1 - Anticancer agent containing vitamin k3 and conjugated linoleic acid - Google Patents

Abstract

An anticancer composition containing vitamin K3 and conjugated linoleic acid is provided to improve side effect caused by vitamin K3 such as inhibition of intercellular signal transfer through gap junction channel, and increase MMP(matrix metalloproteinase)-inhibiting effects compared to use of single vitamin K3 by using vitamin K3 and conjugated linoleic acid. The anticancer composition contains 0.1-50 wt.% of effective ingredients including vitamin K3 and conjugated linoleic acid in a weight ratio of 10:1 to 1:10, and pharmaceutically acceptable carriers, diluents or excipients, wherein the cancer is large intestine cancer, breast cancer or liver cancer; the daily dosage of effective ingredients is 0.1-100 mg/kg; and the composition is formulated as plaster, granule, lotion, liniment, lemonade, aromatic water, powder, syrup, ophthalmic ointment, liquid and solution, aerosol, extract, elixir, ointment, fluid extract, emulsion, suspension, decoction, infusion, ophthalmic solution, tablet, suppository, injection, spirit, cataplasma, capsule, cream, troche, tincture, paste, pill or soft or hard gelatin capsule.

Description

ANTICANCER AGENT CONTAINING VITAMIN K3 AND CONJUGATED LINOLEIC ACID, comprising vitamin X3 and conjugated linoleic acid as active ingredients

1A to 1D are photographs confirming whether intracellular signal transduction is inhibited through a gap binding channel, a is a positive control group, b is a negative control group, and c is a vitamin K ₃ 50 μM + conjugated linoleic acid 5 μg / ml Is a group treated with vitamin K ₃ 50 μM + conjugated linoleic acid 20 μg / ml.

FIG. 2 is a graph quantifying the results of FIG. 1.

Figure 3 confirms the effect of inhibiting MMP activity when treated with a mixture of vitamin K ₃ and conjugated linoleic acid in cancer cell line HT-29, respectively.

The present invention relates to an anticancer composition comprising vitamin K ₃ and conjugated linoleic acid (CLA) as an active ingredient, and more particularly, inhibition of intercellular signal transduction as a side effect of vitamin K ₃ . The present invention relates to an anticancer composition which is prepared by mixing conjugated linoleic acid with vitamin K _{3 in} order to recover and to enhance the inhibitory effect of matrix metalloproteinase.

Drug therapy, surgical therapy, and radiation therapy are mainly performed as a method of treating cancer, and various methods have been attempted. However, drug therapy has inevitably caused side effects due to high doses of drugs for proper integration of anticancer drugs into the lesions. For example, the use of Holmium (HolmiuM), which has been shown to be effective in cancer, to form a complex with chitosan (Milican, Donghwa Pharm. Co., Ltd.) has been shown to significantly increase the anticancer effect, while causing serious side effects. (Watterson. JD et al., J. Urol . 168: 442-445, 2002; Peh, OH et al., Ann. Acad. Med. Singapo re 30: 563-567, 2001).

Matrix metalloproteinase (hereinafter referred to as "MMP") is an enzyme that is important for cancer migration and metastasis, and is a representative enzyme that dissolves extracellular matrix. To date, it has been confirmed that MMP expression is increased in various types of carcinoma. Of these, metalloproteinase 7 (hereinafter “MMP-7”) is known to be most directly associated with cancer metastasis, and has been reported to play an important role in the metastasis of colorectal cancer (Brabletz T et al., AMERICAN JOURNAL OF PATHOLOGY 155 (4): 1033-1038 , 1999; Wilson CL et al., INTERNATIONAL JOURNAL OF BIOCHEMISTRY & CELL BIOLOGY 28 (2): 123-136, 1996; Imai K et al., JOURNAL OF BIOLOGICAL CHEMISTRY 270 (12): 6691-6697, 1995; Imai K et al., BIOCHEMICAL JOURNAL 322: 809-814, 1997; Giambernardi TA et al., MATRIX BIOLOGY 16 (8): 483-496, 1998; Mori M et al. , CANCER 75 (6): 1516-1519 Suppl, 1995; Adachi Y et al., GUT 45 (2): 252-258, 1999; Lynch CC et al., DIFFERENTIATION 70 (9-10): 561-573, 2002; Hasegawa S et al., INTERNATIONAL JOURNAL OF CANCER 76 (6): 812-816, 1998; Chakraborti S et al., MOLECULAR AND CELLULAR BIOCHEMISTRY 253 (1-2): 269-285 2003). Accordingly, a substance capable of inhibiting the activity of MMP may be proposed as an anticancer agent.

On the other hand, while inhibition of intercellular signal transduction through gap junction channels is recognized as an important biochemical indicator of cancer development, agents that induce this process are well known to induce the promotion and progression of cancer ( Eghbali B. et al., Proc. Natl. Acad. Sci. 87: 1328-1331, 1990; Ara C et al., Cell Mol Life Sci. 59 (10): 1758-65, 2002; Evert M et al. , Carcinogenesis 23 (5): 697-703, 2002; Muramatsu A et al., Carcinogenesis 23 (2): 351-8, 2002). Accordingly, development of a substance capable of restoring intercellular signal transduction through a gap binding channel as an anticancer agent may be proposed.

Quinone compounds that are used to treat solid and many cancers (Nutter LM et al., Biochem Pharmacol. , 41 (9): 1283-92, 1991), doxorubicin, has been reported to have side effects that inhibit intercellular signaling through gap junction channels (Kotb A et al., European Journal of Biochemistry, ps01- 0688, 2003).

In addition, vitamin K _{3, which} is known to have a very high anticancer effect, also has a side effect of inhibiting intracellular signal transduction through a gap binding channel.

Thus, vitamin K₃The development of a material that can compensate for the side effect of inhibiting signal transduction between cells through the gap binding channel of the cell is required.

The present invention has been made to overcome the above problems, the side effect of inhibiting the intercellular signal transmission through the gap binding channel by vitamin K ₃ is improved, compared to the use of vitamin K ₃ alone MMP inhibitory effect is enhanced It is an object to provide an anticancer composition.

In order to achieve the above object, the present invention provides an anticancer composition comprising vitamin K ₃ and conjugated linoleic acid as an active ingredient.

Hereinafter, the configuration of the present invention will be described in more detail.

While the present inventors are studying anti-cancer drugs that have secured human safety, conjugated linoleic acid prevents the side effects of inhibiting intracellular signal transduction through the interstitial binding channel of vitamin K ₃ , and at the same time inhibits the progression of cancer. It was confirmed that it can enhance the MMP inhibitory effect of ₃ , to complete the present invention based on this.

The present invention relates to an anticancer composition containing vitamin K ₃ and conjugated linoleic acid as an active ingredient, wherein the conjugated linoleic acid can be purchased as a commercially available compound.

The anticancer composition of the present invention may further include a pharmaceutically acceptable carrier, diluent or excipient in addition to the active ingredient, wherein the content of the active ingredient vitamin K ₃ and conjugated linoleic acid is preferably 0.1 to the total composition weight. 50 wt% is preferred.

Carriers, excipients or diluents which may be used include lactose, dextrose, sucrose, sorbitol, mannitol, xyitol, erythritol, maltitol, starch, acacia rubber, alginate, gelatin, calcium phosphate, calcium silicate, cellulose, methyl cellulose, Microcrystalline cellulose, polyvinylpyrrolidone, water, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate and mineral oil, which may be used one or more. In addition, when the anticancer composition is a drug, it may further include a filler, an anticoagulant, a lubricant, a humectant, a perfume, an emulsifier or a preservative.

In addition, the anticancer composition comprising the vitamin K ₃ and conjugated linoleic acid of the present invention as an active ingredient, when used as a medicament may be an anticancer agent, when used as food, food additives or beverages, various foods, meat , Beverages, chocolate, snacks, confectionery, pizza, ramen, other noodles, gum, ice cream, alcoholic beverages, vitamin complexes, alcohol and other health supplements, but is not necessarily limited thereto.

On the other hand, the ratio of vitamin K ₃ and conjugated linoleic acid contained in the anticancer composition of the present invention may be variously controlled according to the method of using the anticancer composition, the condition of the user, the type of disease, and the severity of the disease, but is preferred. Preferably, the mixing ratio of vitamin K ₃ and conjugated linoleic acid is 10: 1 to 1:10 by weight. However, this may be adjusted according to a therapeutic purpose and a preventive purpose, but is not necessarily limited to the above range. However, if the ratio is out of the range, the anticancer effect may be somewhat insignificant.

On the other hand, the dosage form of the anticancer composition may be in various forms depending on the method of use, and in particular, it is formulated by adopting methods known in the art to provide rapid, sustained or delayed release of the active ingredient after administration to a mammal. It is good.

Examples of specific formulations include PLASTERS, GRANULES lotions, LPTIONS, LINIMENTS, LIMONADES, AROMATIC WATERS, POWDERS, Syrups. ), OPHTALMIC OINTMENTS, LIQUIDS AND SOLUTIONS, AEROSOLS, EXTRACTS, ELIXIRS, OINTMENTS, FLUIDEXTRACTS, Emulsions , SUPESIONS, DECOCTIONS, INFUSIONS, OPHTHALMIC SOLUTIONS, TABLETS, Suppositories (SUPPOSITIORIES), INJECTIONS, SPIRITS, CATAPLSMA , Capsules (CAPSULES), creams (CREAMS), troches (TROCHES), tinks (TINCTURES), pasta (PASTES), pills (PILLS), soft or hard gelatin capsules.

The dosage of the anticancer composition according to the present invention may be determined in consideration of the method of administration, the age, sex and weight of the recipient, and the severity of the disease. As an example, the anticancer composition of the present invention may be administered at least once at 0.1 to 100 mg / kg (body weight) per day based on the active ingredient. However, the above dosage is only one example to illustrate and is not necessarily limited to the above range.

Anti-cancer composition containing the vitamin K _3, and conjugated linoleic acid in accordance with the present invention as an active ingredient sikimyeo significantly improve cell signaling inhibitory side effect through the gap junction of vitamin K _3, the MMP inhibitory effect as compared to vitamin K ₃ alone great. Therefore, the anticancer composition of the present invention can be used for the purpose of preventing and treating cancer progression and metastasis in the early stage of cancer development, and also can be applied to various cancers, such as colon cancer, breast cancer and liver cancer.

Hereinafter, the configuration of the present invention will be described in more detail with reference to the following examples, but the scope of the present invention is not limited to the following examples.

Meanwhile, in the following examples solids and solid mixtures, liquids and liquids, and percentages for liquids and solids are based on weight / weight, volume / volume and weight / volume, respectively, and all reactions were performed at room temperature unless otherwise noted. .

Example 1: Verification of the effect of restoring the intercellular signal transduction mechanism through the gap binding channel by conjugated linoleic acid

To determine whether conjugated linoleic acid has the effect of restoring intercellular signal transduction through interstitial binding channels inhibited by vitamin K ₃ , the SL / DT (Scrape Loading / Dye Transfer assay; Upham BL et al., Carcinogenesis, 18: 37-42, 1997).

WB-F344 cells (University of Michigan, USA) were prepared using DMEM medium supplemented with 10% FBS, 100 IU / mL of penicillin and 100 μg / mL of streptomycin, and a 5% CO ₂ , 37 ° C. incubator (Forma Scientific Co. , Marjetta, OH, USA). The medium compositions used for cell culture were all used by GIBCO BRL (Grand Island, NY, USA).

The cultured cells were dispensed at 1 x 10 ⁴ per ml in a 2 ml plastic culture plate and incubated for about 48 hours. After cultivation, the cells were cultured in each culture plate with vitamin K ₃ 50 μM + conjugated linol. 5 μg / ml of lane acid, 50 μM of vitamin K ₃ + 20 μg / ml of conjugated linoleic acid were treated. Positive control group was treated with culture solution, negative control group was treated with vitamin K ₃ 50 μM. After 1 hour staining with lucifer yellow, the degree of intercellular signaling through the gap binding channel was observed by confocal microscopy (BioRad, Hercules, CA, USA).

1A to 1D are photographs confirming whether the intercellular signal transduction is inhibited through the gap binding channel, a is a positive control group, b is a negative control group, and c is a vitamin K ₃ 50 μM + 5 μg / ml conjugated linoleic acid. And d is a group treated with vitamin K ₃ 50 μM + conjugated linoleic acid 20 μg / ml.

As shown in FIG. 1, when the cells are treated with vitamin K ₃ alone, intercellular signal transduction is rapidly inhibited through the gap binding channel, but when the conjugated linoleic acid is treated together, signal transduction inhibition is inhibited. It works.

In addition, in order to more clearly confirm the recovery degree of intercellular signal transduction inhibition through interstitial binding channels induced by vitamin K ₃ , the number of cells in which intercellular signal transduction was measured was shown in FIG. 2. In FIG. 2, it was confirmed that the intercellular signal transduction, which was inhibited during vitamin K ₃ treatment, was restored to about 75% due to the conjugated linoleic acid treatment.

Therefore, the addition of conjugated linoleic acid may have an effect of preventing intercellular signal transduction through a gap binding channel which is a side effect of conjugated linoleic acid.

Example 2: Vitamin K ₃ MMP Inhibitory Effect and MMP Inhibitory Effect of Conjugated Linoleic Acid Additives

HT-29 cells, which are human colon cancer cells, were cultured for 24 hours, washed with PBS, incubated for 2 hours with serum-free media, and treated with a concentration of vitamin K ₃ and conjugated linoleic acid. Further incubation for 48 hours. Culture supernatants were collected and centrifuged (vision, DN-5500) and protein content was determined using a DC Assay Kit [Bio-Rad Corp., Richmond, CA, USA]. About 15 μg of protein was extracted from each protein extract by electrophoresis in 12.5% SDS-PAGE. MMP-7 was reacted with a monoclonal antibody [Chemicon, Temecula, CA, USA] and then detected using an ECL kit [Amersham, Life Science, Denver, USA].

Figure 3 shows the MMP inhibitory effect of the mixture of vitamin K ₃ and conjugated linoleic acid, lane 1 is the untreated control, lane 2 is vitamin K ₃ 50 μM treated group, lane 3 is vitamin K ₃ 50 μM + conjugated Linoleic acid 20 μg / ml treatment group.

As shown in FIG. 3, the effect of inhibiting the activity of MMP-7 by conjugated linoleic acid is further enhanced by addition of conjugated linoleic acid.

Example 3: Anticancer Agent Preparation

3-1. Powder

1 g of lactose was mixed with 2 g of a mixture of vitamin K ₃ / conjugated linoleic acid (10: 1 weight ratio) and filled into an airtight cloth to prepare a powder.

3-2. refine

Tablets were prepared by mixing 100 mg of vitamin K ₃ / conjugated linoleic acid (10: 1 weight ratio), 100 mg of corn starch, 100 mg of lactose, and 2 mg of magnesium stearate, followed by compression according to a conventional tablet preparation method.

3-3. Capsule

A capsule was prepared by mixing 100 mg of vitamin K ₃ / conjugated linoleic acid (10: 1 weight ratio), 100 mg of corn starch, 100 mg of lactose, and 2 mg of magnesium stearate, and then filling the gelatine capsule.

3-4. Injection

Injectable solution was prepared by adding a suitable amount of distilled water for injection to 100 mg of a mixture of vitamin K ₃ / conjugated linoleic acid (10: 1 weight ratio), adjusting the pH to about 7.5, and then filling and sterilizing a 2 ml ampoule. It was.

Example 4: Functional Food Preparation

4-1. Wire

Brown rice, barley, glutinous rice, and yulmu were alphatized by a known method, and then dried and roasted to prepare a powder having a particle size of 60 mesh using a grinder. Black beans, black sesame seeds and perilla were each steamed and dried in a known manner, then roasted and ground to prepare a powder having a particle size of 60 mesh.

Then, 30% by weight brown rice, 15% by weight barley, 20% by weight barley, 9% by weight glutinous rice, 7% by weight perilla, 8% by weight black soybean, 7% by weight black sesame, vitamin K ₃ / conjugated linoleic acid (10: 1 Weight ratio) 3% by weight, ganoderma lucidum 0.5% by weight and sulfur by weight 0.5% by weight was prepared to prepare a wire.

4-2. Chewing gum

Chewing gum was prepared in a conventional manner by combining 20% by weight of gum base, 76.9% by weight of sugar, 1% by weight of perfume, 2% by weight of water, and 0.1% by weight of vitamin K ₃ / conjugated linoleic acid (10: 1 weight ratio). .

4-3. candy

Candy was prepared in a conventional manner by combining 0.1% by weight of vitamin K ₃ / conjugated linoleic acid (10: 1 weight ratio).

4-4. Biscuits

Force 1st class 25.59 wt%, 1st class gravity 22.22 wt%, white sugar 4.80 wt%, salt 0.73 wt%, glucose 0.78 wt%, palm shortening 11.78 wt%, ammonium 1.54 wt%, sodium bicarbonate 0.17 wt%, sodium bisulfite 0.16 wt %, Rice flour 1.45 wt%, Vitamin B₁0.0001 wt%, Vitamin B20.0001 wt%, Milk flavor 0.04 wt%, Water 20.6998 wt%, Whole milk powder 1.16 wt%, Substitute milk powder 0.29 wt%, Calcium phosphate 0.03 wt% , Biscuits were prepared in a conventional manner by combining 0.29 wt% of spraying salt, 7.27 wt% of spray oil, and 1 wt% of Vitamin K ₃ / conjugated linoleic acid (10: 1 weight ratio).

4-5. Health drink

0.26 wt% honey, 0.0002 wt% thioctoamide, 0.0004 wt% nicotinamide, 0.0001 wt% sodium riboflavinate, 0.0001 wt% pyridoxine hydrochloride, 0.001 wt% inositol, 0.002 wt% ortic acid, 98.7362 wt% water and vitamin K _A health beverage was prepared in a conventional manner by combining 3% by weight of conjugated linoleic acid (10: 1 weight ratio).

4-6. sausage

Pork 65.18%, broiler 25%, starch 3.5%, soy protein 1.7%, salt 1.62%, glucose 0.5% and glycerin 1.5% and vitamin K ₃ / conjugated linoleic acid (10: 1 weight ratio) ) 1% by weight of sausage was prepared by the conventional method.

4-7. Health Supplement

55% by weight of spirulina, 10% by weight of guar gum enzyme digestion, 0.01% by weight of vitamin B₁ hydrochloride, 0.01% by weight of vitamin B6 hydrochloride, 0.23% by weight of DL-methionine, 0.7% by weight of magnesium stearate, 22.2% by weight of lactose and 1.85% by weight of corn starch And 10% by weight of vitamin K ₃ / conjugated linoleic acid (10: 1 weight ratio) to prepare a tablet-type health supplement in a conventional manner.

4-8. mainstream

0.5% vitamin K ₃ / conjugated linoleic acid (10: 1 weight ratio) is mixed with soju, beer, liquor or fruit liquor to make an emulsion, and then centrifuged at 7,000 rpm for 15 minutes in a vacuum or 9,000 with a high speed mixer. Mixing at rpm produced a liquor containing vitamin K ₃ / conjugated linoleic acid.

As it is seen from the foregoing, thereby enhancing the conjugated linoleic acid in combination with vitamin K ₃ processing, prevent signal between cells via gap junction channels in the side effects of vitamin K ₃ passes effectively suppressed, and the MMP inhibitory effect of the present invention. Therefore, the anticancer composition having conjugated linoleic acid added to vitamin K ₃ may be an ideal anticancer agent having a good anti-cancer effect and minimized side effects.

Claims (5)

An anticancer composition comprising vitamin K ₃ and conjugated linoleic acid as an active ingredient. According to claim 1, The content ratio of the total composition of vitamin K ₃ and conjugated linoleic acid contained as an active ingredient, Anticancer composition, characterized in that 0.1 to 50% by weight relative to the total composition weight. The mixing ratio of vitamin K ₃ and conjugated linoleic acid according to claim 2, Anticancer composition, characterized in that from 10: 1 to 1:10 weight ratio. delete The method of claim 1, wherein the cancer, Anticancer composition, characterized in that any one selected from colon cancer, breast cancer, liver cancer.

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