KR20140141505A - Pharmaceutical compositon for inhibiting fibrosarcoma metastases containing ketotifen - Google Patents

Abstract

The present invention relates to a pharmaceutical composition containing ketotifen as an active ingredient for inhibiting fibro sarcoma metastasis. Ketotifen of the present invention has an outstanding effect of preventing metastasis by inhibiting migration of cancer cells in fibrosarcoma, so ketotifen can be used as a pharmaceutical preparation for inhibiting fibrosarcoma metastasis or for anticancer aids.

Description

[0001] The present invention relates to a pharmaceutical composition for inhibiting fibrosarcoma metastasis, which contains ketotifen as an active ingredient.

The present invention relates to a pharmaceutical composition for inhibiting fibrous sarcoma cancer metastasis containing kettifen as an active ingredient.

Cancer cells mean that cancer cells move from one organ or native site to another organ or to other parts. It is known that cancer metastasis is mainly caused by malignant cancer cells. Most of the metastatic cancer cells come out of the primary cancer and penetrate into the lymphoid or vascular system, and then circulate through the tissue or blood vessels and then penetrate and grow in other tissues. Cancer is a typical feature of malignant tumors, and mortality from malignant cancer is known to account for 90% of all cancer deaths (Weigelt et al., 2005).

An important role involved in metastasis is angiogenesis, cell migration and epithelial-mesenchymal transition (EMT). To meet the metabolic needs that grow as cancer cells grow, they migrate from the epithelial state to the mesenchymal state for movement, which is called angiogenesis. Through this process, newly formed blood vessels are known to act as a pathway for cancer cells to leave the cancer tissue and enter the circulation system of the human body (intravasation). In addition, cancer cells that have secured mobility may indirectly enter the vascular circulation system using the lymphatic system.

Cancer cells must survive in the circulatory system until growth is arrested in new organs. At restrained, cancer cells are ejected from the circulatory system into surrounding tissues, and at new sites, the cells continue to grow and are known to form pre-angiogenic micrometastases. Thus, the growth of cancer cells is maintained by angiogenesis, resulting in the formation of an observable cancer. Therefore, in order for cancer cells to transfer, cancer cells must necessarily move.

The treatment of cancer is mainly surgical operation, chemotherapy, radiotherapy etc, but most of them have a significant effect. However, some surviving cancer cells migrate in the patient's body and are metastasized, so that the cancer is often recurred, so that the suffering of the patient is not reduced. Therefore, there is a need to develop a new therapeutic agent which can alleviate the suffering of cancer patients and alleviate the anxiety of cancer metastasis. Accordingly, anticancer agents have been studied focusing on the treatment of advanced cancer and the remarkable decrease in cancer recurrence rate.

In the case of drugs that inhibit cancer metastasis, it is important to identify appropriate markers for metastatic cancer. Studies on biomarkers have been actively conducted to date, There are still many difficulties in doing so. Particularly, there is a demand for the development of a useful anti-cancer agent that can suppress cancer metastasis after surgery.

On the other hand, Ketotifen (4- (methyl- 4-piperidylidene) -4Hbenzo [4,5] cyclohepta [1,2-b] thiophen-10 It has been commonly used as an antianaphylactic agent. Ketotifen has been used primarily as an oral preparation to prevent allergic diseases by directly inhibiting the release of allergen-inducing mediators from mast cells. Ketotifen is also formulated into syrups or capsules and is used to treat asthma, allergic bronchitis, allergic conjunctivitis, allergic dermatitis, atopic dermatitis and the like, and is also used as an eye drop for ophthalmic diseases and as a topical application for external application .

As described above, various pharmacological effects of ketotifen are known, but the effect of ketotifen on the inhibition of metastasis of fibre sarcoma has not been reported, and there have been no studies on this.

The inventors of the present invention investigated drugs that have various compounds and mechanisms that are unclear in order to find a new substance that inhibits fibrosarcoma metastasis. Ketotifen inhibits the migration of cancer cells in fibrosarcoma, And thus the present invention has been completed.

It is an object of the present invention to provide a pharmaceutical composition for inhibiting fibrosarcoma cancer metastasis containing kettifen as an active ingredient.

Another object of the present invention is to provide a pharmaceutical composition for adjuvant therapy comprising ketotifen as an active ingredient.

It is still another object of the present invention to provide an anticancer-aid food composition containing ketotifen as an active ingredient.

The present invention provides a pharmaceutical composition for inhibiting fibrous sarcoma cancer metastasis containing kettifen as an active ingredient.

The present invention also provides a pharmaceutical composition for adjuvant therapy comprising ketotifen as an active ingredient.

The present invention also provides an anticancer-aid food composition containing kettifen as an active ingredient.

Ketotifene of the present invention is useful as a pharmaceutical preparation for inhibiting metastasis of fibrosarcoma or for anticancer assistant because it inhibits the movement of cancer cells in fibrosarcoma and is excellent in suppressing cancer metastasis.

Brief Description of the Drawings Fig. 1 is a graph showing the effect of inhibiting cell migration in keratothelin fibro sarcoma according to the present invention. Fig.
FIG. 2 is a graph showing the cell infiltration inhibitory effect of kestotin in fibro sarcoma according to the present invention.
FIG. 3 is a graph showing the effect of ketotifen on the expression of cell migration-related proteins according to the present invention.
FIG. 4 is a graph showing the effect of kettotin according to the present invention on the expression and activity of MMP-9. FIG.

The present invention provides a pharmaceutical composition for inhibiting fibrous sarcoma cancer metastasis containing kettifen as an active ingredient.

The present invention also provides a pharmaceutical composition for adjuvant therapy comprising ketotifen as an active ingredient.

The present invention also provides an anticancer-aid food composition containing kettifen as an active ingredient.

Hereinafter, the present invention will be described in detail.

Ketotifene of the present invention is useful as a pharmaceutical preparation for inhibiting metastasis of fibrosarcoma or for anticancer assistant because it inhibits the movement of cancer cells in fibrosarcoma and is excellent in suppressing cancer metastasis.

It is also preferable that ketotifen of the present invention is used in combination with an anticancer agent.

The anticancer agent may be selected from the group consisting of Busulfan, Chlorambucil, Cyclophosphamide, Melphalan, Nitrogen Mustard, 1,3-bis (2-chloroethyl) ) -1 nitrosourea BCNU, l- (2-Chloroethyl) -3-Cyclohexyl-1-Nitrosourea (CCNU), 1- (2-chloroethyl) -3- (4-methyl- cyclohexyl) 1-nitrosourea (MeCCNU), cisplatin, ifosfamide, cytarabine, fluouracil (5-FU), mercaptopurine (6-MP), methotrexate Methotrexate (MTX), Actinomycin-D, Bleomycin, Daunorubicin, Adriamycin, Mitomycin-C, Vinblastine ), Vincristine (Oncovin), Vindesine, Etoposide (VP-16), L-asparaginase, Dacarbazine (DTIC) and hydroxyurea (Hyd roxyurea), but the present invention is not limited thereto.

The composition of the present invention may contain one or more known active ingredients having an effect of preventing or treating fibrosarcoma cancer metastasis together with ketotifen.

The compositions of the present invention may further comprise suitable carriers, excipients and diluents conventionally used in the manufacture of pharmaceutical compositions. In addition, it can be formulated in the form of powders, granules, tablets, capsules, suspensions, emulsions, oral formulations such as syrups and aerosols, external preparations, suppositories and sterilized injection solutions according to a conventional method. Suitable formulations known in the art are preferably those as disclosed in Remington ' s Pharmaceutical Science, recently, Mack Publishing Company, Easton PA. Examples of carriers, excipients and diluents which may be included include lactose, dextrose, sucrose, sorbitol, mannitol, xylitol, erythritol, maltitol, starch, acacia rubber, alginate, gelatin, calcium phosphate, calcium silicate, cellulose, methylcellulose, Cellulose, polyvinylpyrrolidone, water, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate, and mineral oil. When the composition is formulated, it is prepared using a diluent such as a filler, an extender, a binder, a wetting agent, a disintegrant, a surfactant, or an excipient usually used. Solid formulations for oral administration include tablets, pills, powders, granules, capsules and the like, which may contain at least one excipient such as starch, calcium carbonate, sucrose, lactose, Gelatin and the like. In addition to simple excipients, lubricants such as magnesium stearate and talc may also be used. Examples of the liquid preparation for oral administration include suspensions, solutions, emulsions, and syrups. In addition to water and liquid paraffin, simple diluents commonly used, various excipients such as wetting agents, sweeteners, fragrances, preservatives and the like may be included . Formulations for parenteral administration include sterilized aqueous solutions, non-aqueous solutions, suspensions, emulsions, freeze-dried preparations, and suppositories. Examples of the suspending agent include propylene glycol, polyethylene glycol, vegetable oil such as olive oil, injectable ester such as ethyl oleate, and the like. Examples of the suppository base include witepsol, macrogol, tween 61, cacao butter, laurin, glycerogelatin and the like.

The term "administering" as used herein is meant to provide any desired composition of the invention to a subject in any suitable manner.

The preferred dosage of the pharmaceutical composition of the present invention varies depending on the condition and the weight of the individual, the degree of disease, the type of drug, the route of administration and the period of time, but can be appropriately selected by those skilled in the art. For the desired effect, the ketotifen of the present invention may be administered in an amount of 1 mg / kg to 10000 mg / kg per day, and may be administered once a day or divided into several doses.

The pharmaceutical composition of the present invention may be administered to a subject in various routes. All modes of administration may be expected, for example, by oral, rectal or intravenous, intramuscular, subcutaneous, intra-uterine dural or intracerebral injection.

The composition of the present invention can be used alone or in combination with methods using surgery, radiation therapy, hormone therapy, chemotherapy and biological response modifiers for the prevention and treatment of metastasis of fibrosarcoma cancer.

The composition of the present invention may be added to a health functional food for the purpose of inhibiting fibrosarcoma cancer metastasis. When ketotiphen of the present invention is used as a food additive, the ketotifen may be added as it is, or may be used together with other food or food ingredients, and suitably used according to a conventional method. The amount of the active ingredient to be mixed can be suitably determined according to the intended use (prevention, health or therapeutic treatment). Generally, in the production of foods or beverages, ketotifen of the present invention is added in an amount of not more than 15% by weight, preferably not more than 10% by weight based on the raw material. However, in the case of long-term intake for the purpose of health and hygiene or for the purpose of controlling health, the amount may be less than the above range, and since there is no problem in terms of safety, the active ingredient may be used in an amount exceeding the above range.

There is no particular limitation on the kind of the food. Examples of foods to which the above substances can be added include dairy products including meat, sausage, bread, chocolate, candy, snacks, confectionery, pizza, ramen and other noodles, gums, ice cream, various soups, drinks, tea, Alcoholic beverages, and vitamin complexes, all of which include health foods in a conventional sense.

The health beverage composition of the present invention may contain various flavors or natural carbohydrates as an additional ingredient such as ordinary beverages. The natural carbohydrates may be monosaccharides such as glucose and fructose, disaccharides such as maltose and sucrose, natural sweeteners such as dextrin and cyclodextrin, synthetic sweeteners such as saccharine and aspartame, and the like. The ratio of the natural carbohydrate is generally about 0.01 to 10 g, preferably about 0.01 to 0.1 g per 100 ml of the composition of the present invention.

In addition to the above, the composition of the present invention may further contain various nutrients, vitamins, electrolytes, flavors, colorants, pectic acid and salts thereof, alginic acid and salts thereof, organic acids, protective colloid thickeners, pH adjusters, stabilizers, preservatives, glycerin, A carbonating agent used in a carbonated beverage, and the like. In addition, the composition of the present invention may comprise flesh for the production of natural fruit juices, fruit juice drinks and vegetable drinks. These components may be used independently or in combination. Although the ratio of such additives is not critical, it is generally selected in the range of 0.01 to 0.1 parts by weight per 100 parts by weight of the composition of the present invention.

Hereinafter, preferred embodiments and examples of the present invention will be described in order to facilitate understanding of the present invention. However, the following examples and preparative examples are provided only for the purpose of easier understanding of the present invention, and the present invention is not limited by the examples and preparation examples.

Example 1. Measurement of inhibitory activity of ketotifene against fibrosarcoma cell migration

In order to examine the inhibitory activity of ketotifen inhibition of fibrosarcoma metastasis of the present invention, the inhibition activity of ketotifen against fibrosarcoma cancer cells was measured.

1-1. Preparation and culture of HT-1080 cells

The fibrosarcoma cancer cell line HT-1080 cells were cultured in RPMI 1640 (Weljin) medium containing 10% fetal bovine serum (FBS) and L-glutamine. 1% antibiotic (penicillin-G 100 units / ml / streptomycin 100 ug / ml) was added to the medium. Cell cultures were performed in an incubator (Forma 3111, formma Scientific.Lnc. USA) containing carbon dioxide at a temperature of 37 ° C and a concentration of 5%.

1-2. Measurement of cell migration inhibitory activity of ketotifen

The cell migration inhibitory activity of HT-1080 cells prepared in Example 1-1 was measured.

Specifically, for the migration experiment, a multi-well chamber of Neuro Probe coated with 10 μg / ml of fibronectin as a chemoattractant was used (Park et al., 2011) . Ketotifene of the present invention was dissolved in DMSO and pretreated for each concentration beforehand. HT-1080 cells were suspended in DMEM medium at a density of 1.1 × 10 ⁷ cells / ml, and 25 μl of them were taken in an upper chamber of a multichamber. Thereafter, an 8 μm polyhydrocarbon filter was placed in the middle well, DMEM medium containing 3% serum was placed in the lower well, and then cultured at 37 ° C. for 4 hours. After completion of the incubation, the non-migrating cells on the upper surface of the filter were wiped off thoroughly, and the cells migrating on the lower surface were stained using a Diff-quick dye. Five randomly selected cells were selected from the stained cells, and each cell in the selected region was identified by a microscope with a magnification of 400X and counted. The results are shown in Fig.

As shown in FIG. 1, the kettotin of the present invention significantly inhibited the migration of HT-1080 cells, a fibrosarcoma cancer cell line, at a concentration of 25 μM.

Therefore, it was confirmed that the ketotifen of the present invention significantly inhibited the migration of HT-1080 cells, which is a cell sarcoma cancer cell line, to thereby exert an excellent effect of inhibiting metastasis of fibrosarcoma cancer.

Example 2. Measurement of inhibitory activity of ketotifene against HT-1080 cells

Matrigel (TM) invasion assay was performed on the HT-1080 cells prepared in Example 1-1 to confirm the effect of ketotifen on the infiltration of HT-1080 cells, a fibrosarcoma cancer cell line. The inside of the transwell insert was coated with 500 μg / ml of Matrigel to induce invasion. Twenty nM of TPA (12-O-tetradecanoylphorbol-13 acetate) inducing invasion and various concentrations of ketotifen were treated with the HT-1080 cell line and cultured for 1 hour. 1 x 10 ⁴ cells per well were placed in a container, cultured in an incubator for 24 hours, and the number of cells moved downward was counted under a 200-fold microscope. The results are shown in Fig.

As shown in Fig. 2, it was confirmed that ketotifene inhibits the infiltration of HT-1080 cells in a concentration-dependent manner.

Example 3. Measurement of effect of ketotifen on the expression of cell migration-related proteins

Western blotting was performed to determine the effect of HT-1080 on cell migration-related proteins in ketotifen treatment. TPA, and ketotifene, respectively, on the expression of cdc42 and Rho, the cell migration-related proteins of HT-1080, and the results are shown in Fig.

As shown in FIG. 3, the expression of cdc42 and Rho was increased by TPA treatment, but the expression of these proteins was decreased when keto-typhine was treated.

Example 4. Measurement of the effect of kettifen on the expression and activity of cell infiltrating MMP-9 (Matrix metallopeptidase 9)

MMP-9 (Matrix metallopeptidase 9) is a gene involved in invasion and metastasis of cancer, and the effect of ketotifen on the expression and activity of MMP-9 was measured.

The HT-1080 cell line was treated with TPA and ketotifene, respectively, and the proteins extracted from the cells were electrophoresed to confirm the MMP-9 band. The results are shown in FIG.

As shown in FIG. 4A, an increase in MMP-9 was observed upon TPA treatment, and it was confirmed that treatment with ketotifen inhibited the expression of MMP-9 in a concentration-dependent manner.

Also, as shown in FIG. 4B, it was confirmed that MMP-9 increased in secretion by TPA was reduced by treatment with ketotifen. This indicates that ketotifene inhibits the activity of MMP-9 through inhibition of MMP-9 expression and has an activity of inhibiting invasion and metastasis of cancer cells.

Hereinafter, the pharmaceutical composition of the present invention and the preparation example of the food composition will be described, but the present invention is not intended to be limited but is specifically described.

Formulation Example 1. Preparation of pharmaceutical preparations

1. Manufacturing of powder

Ketotypen 20 mg Lactose 100 mg Talc 10 mg

The above components are mixed and filled in airtight bags to prepare powders.

2. Preparation of tablets

Ketotypen 10 mg Corn starch 100 mg Lactose 100 mg Magnesium stearate 2 mg

After mixing the above components, tablets are prepared by tableting according to the usual preparation method of tablets.

3. Preparation of capsules

Ketotypen 10 mg Crystalline cellulose 3 mg Lactose 14.8 mg Magnesium stearate 0.2 mg

The above components are mixed according to a conventional capsule preparation method and filled in gelatin capsules to prepare capsules.

4. Preparation of injections

Ketotypen 10 mg Mannitol 180 mg Sterile sterilized water for injection 2974 mg Na ₂ HPO ₄ 2H ₂ O 26 mg

(2 ml) per 1 ampoule in accordance with the usual injection preparation method.

5. Manufacture of liquids

Ketotypen 20 mg Isomer 10 g Mannitol 5 g Purified water Suitable amount

Each component was added and dissolved in purified water according to the usual liquid preparation method, and the lemon flavor was added in an appropriate amount. Then, the above components were mixed, and purified water was added thereto. The whole was added with purified water to adjust the total volume to 100 ml, And sterilized to prepare a liquid preparation.

Formulation Example 2. Food preparation

2-1. Manufacture of health food

Ketotypen 100 mg Vitamin mixture Suitable amount Vitamin A acetate 70 μg Vitamin E 1.0 mg Vitamin B1 0.13 mg Vitamin B2 0.15 mg Vitamin B6 0.5 mg Vitamin B12 0.2 μg Vitamin C 10 mg Biotin 10 μg Nicotinic acid amide 1.7 mg Folic acid 50 μg Calcium pantothenate 0.5 mg Mineral mixture Suitable amount Ferrous sulfate 1.75 mg Zinc oxide 0.82 mg Magnesium carbonate 25.3 mg Potassium monophosphate 15 mg Dicalcium phosphate 55 mg Potassium citrate 90 mg Calcium carbonate 100 mg Magnesium chloride 24.8 mg

Although the composition ratio of the above-mentioned vitamin and mineral mixture is comparatively mixed with a composition suitable for health food as a preferred embodiment, the compounding ratio may be arbitrarily modified, and the above ingredients are mixed according to a conventional method for producing healthy foods , Granules can be prepared and used in the manufacture of health food compositions according to conventional methods.

2-2. Manufacture of health drinks

Ketotypen 100 mg Vitamin C 15 g Vitamin E (powder) 100 g Iron lactate 19.75 g Zinc oxide 3.5 g Nicotinic acid amide 3.5 g Vitamin A 0.2 g Vitamin B1 0.25 g Vitamin B2 0.3 g water Suitable amount

The above components were mixed according to a conventional health drink manufacturing method, and the mixture was stirred and heated at 85 DEG C for about 1 hour. The solution thus prepared was filtered and sterilized in a sterilized 2 liter container, It is used in the production of the health beverage composition of the invention.

Although the compositional ratio is relatively mixed with a component suitable for a favorite drink, it is also possible to arbitrarily modify the compounding ratio according to the regional or national preference such as the demand class, the demanding country, and the use purpose.

Claims (5)

A pharmaceutical composition for inhibiting fibrous sarcoma cancer metastasis, comprising ketotifen as an active ingredient. A pharmaceutical composition for assisting in anti-cancer, comprising kettotin as an active ingredient. The pharmaceutical composition according to claim 2, wherein the ketotifen is used in combination with an anticancer agent. The pharmaceutical composition according to claim 3, wherein the anticancer agent is selected from the group consisting of Busulfan, Chlorambucil, Cyclophosphamide, Melphalan, Nitrogen Mustard, Carmustine (1,3 -bis (2-chloroethyl) -1 nitrosourea, BCNU, l- (2-Chloroethyl) -3-Cyclohexyl-1-nitrosourea (CCNU) 4-methyl-cyclohexyl-1-nitrosourea, MeCCNU, Cisplatin, Ifosfamide, Cytarabine, Fluouracil 5-FU, Mercaptopurine 6 Methotrexate (MTX), Actinomycin-D, Bleomycin, Daunorubicin, Adriamycin, Mitomycin-C, Vincristine; Oncovin; Vindesine; Etoposide (VP-16); L-asparaginase; Dacarbazine; DTIC) and Ha DE hydroxy urea (Hydroxyurea), characterized in that the at least one anticancer agent selected from the group consisting of anti-cancer pharmaceutical composition for the secondary. A composition for supporting anticancer cancer, comprising ketotifen as an active ingredient.

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