JP6742123B2 - Cancer cell migration inhibitor - Google Patents

Description

The present invention relates to a cancer cell migration inhibitor and a method for suppressing cancer cell migration or cancer metastasis using the cancer cell migration inhibitor.

Currently, cancer is the leading cause of death, and the number of patients is increasing year by year. Among them, an extract derived from a natural product having a cancer cell growth inhibitory effect has been found, and since it is highly safe as compared with conventional anticancer agents, a cell growth inhibitor with few side effects and an anti-cancer agent containing these components. It is expected to provide a tumor drug (also called an anti-cancer drug or an anti-cancer drug) and a health food for preventing cancer.

In addition, pets such as dogs tend to be considered to be a member of the family in recent years, and a 2001 survey found that 64% of all people thought that "pets are also members of the family" It is close to humans. On the other hand, neoplastic diseases are increasing as the average life span of pets is extended. In particular, dogs and cats have a high mortality rate due to cancer in old age, and depending on the breed of dog or cat, the incidence of cancer may be higher than that of humans. Similar to humans, cancer treatment methods include surgery, chemotherapy, drug therapy, radiation therapy, and immunotherapy. However, like humans, pets rarely show symptoms in the early stages even if they have cancer, and in many cases, pets often show symptoms such as anorexia and swelling of lymphoma at last. Further, even if the cancer is removed by surgery or the like, the possibility of cancer metastasis cannot be ruled out, so it is necessary to administer an anticancer drug or perform regular examinations. However, since it is not possible to recognize the importance like human beings, the administration of anti-cancer agents and the periodic inspection itself impose a considerable burden on pets, which also causes stress to the owner.

Therefore, there is a need for a drug that can suppress metastasis after cancer treatment. For example, Patent Document 1 reports a cancer cell migration inhibitor containing, as an active ingredient, a short double-stranded oligonucleotide having an RNA interference action on MAST205 (serine threonine kinase).

JP, 2008-308407, A

The cancer cell migration inhibitor described in Patent Document 1 is a nucleic acid molecule. However, in recent years, nucleic acid molecules have not been generally used due to concerns about their safety, and rather, “non-recombinant” is displayed in foods, and their use tends to be refrained.

The present invention has been made in view of the above circumstances, and a highly safe and highly safe cancer cell migration inhibitor that can substitute for a conventionally known cancer cell migration inhibitor and cancer cell migration using the cancer cell migration inhibitor or It is an object to provide a method for suppressing cancer metastasis.

In order to solve the above-mentioned problems, the inventors of the present invention have conducted intensive studies on the cancer cell migration inhibitory effect using extracts of various plants which have been conventionally recognized as safe. As a result, they have found that the combined use of a rosemary extract and a green tea extract at a specific mixing ratio can significantly improve the cancer cell migration inhibitory effect, and completed the present invention.

That is, the above-mentioned object can be achieved by a cancer cell migration inhibitor containing a green tea extract and a rosemary extract in a weight ratio of 10:1 to 1:10 (mixing weight ratio of green tea extract:rosemary extract). ..

Further, the above-mentioned object comprises administering the green tea extract and the rosemary extract in a weight ratio of 10:1 to 1:10 (mixing weight ratio of the green tea extract:rosemary extract), which is used for the migration of cancer cells. It can also be achieved by a suppression method.

According to the present invention, there is provided a highly safe cancer cell migration inhibitor (composition for suppressing cancer cell migration or cancer metastasis), which can substitute for a conventionally known cancer cell migration inhibitor and is highly safe.

It is a photograph which shows the colon cancer cell migration suppression effect by the combined use of a green tea extract and a rosemary extract. It is a graph which shows the colon cancer cell migration suppression effect by the combined use of a green tea extract and a rosemary extract.

Embodiments of the present invention will be described below. The present invention is not limited to the following embodiments. In the present specification, “X to Y” indicating a range includes X and Y, and means “X or more and Y or less”. Unless otherwise specified, operations and measurements of physical properties are performed at room temperature (20 to 25° C.)/relative humidity of 40 to 50% RH.

(Cancer cell migration inhibitor and method for suppressing cancer cell migration)
The cancer cell migration inhibitor of the present invention contains a green tea extract and a rosemary extract in a weight ratio of 10:1 to 1:10 (mixed green tea extract:rosemary extract weight ratio).

In addition, the method for suppressing migration of cancer cells of the present invention uses a green tea extract and a rosemary extract as cancer cells at a weight ratio of 10:1 to 1:10 (green tea extract:rosemary extract mixed weight ratio). Administering to a patient in need of inhibiting migration.

In the previous application (PCT/JP2015/085680), the inventors of the present invention have shown that a composition containing a rosemary extract and a green tea extract in a specific mixing ratio exhibits a remarkable inhibitory effect on cancer cell growth. I found it. As a result of further intensive studies on this composition, the present inventors have found that the composition can synergistically suppress or inhibit the migration of cancer cells. In addition, it is based on the finding in the previous application (PCT/JP2015/085680) that the composition of the above-mentioned specific composition exhibits a significant effect in suppressing the growth of cancer cells. On the other hand, the present invention is based on the finding that it suppresses or inhibits the migration of cancer cells (hereinafter, also simply referred to as “suppression/inhibition”). Here, “cell migration” or “cell migration” means that cells move from their original position to another position due to their motility. By contrast, "proliferation of cells" or "cell proliferation" means that the cells increase their number in situ. Therefore, “cell migration” or “cell migration” and “cell growth” or “cell growth” have completely different meanings, and their development mechanisms are significantly different. Therefore, the technical idea according to the present invention that the composition of the present invention having a specific composition can suppress the migration of cancer cells is the technology according to the previous application that the composition having the same composition as the above can suppress the growth of cancer cells. It is different from the idea, and in the previous application, even if the mixture of the rosemary extract and the green tea extract had the inhibitory effect on the cancer cell growth in the same mixing ratio as the present invention, It does not always have an inhibitory effect on cell migration.

As described above, the cancer cell migration inhibitor of the present invention can suppress or inhibit the migration of cancer cells. Here, the migration of cancer cells is a phenomenon in which cancer cells actively move and also occurs in cancer metastasis, which is a major obstacle to cancer treatment. Therefore, the cancer cell migration inhibitor of the present invention also suppresses/inhibits the phenomenon (metastasis) of migrating (migrating) from a place where a cancer cell has occurred (primary focus) and forming a cancer or tumor again at a remote site. It is thought to be possible. Therefore, the cancer cell migration inhibitor of the present invention is also expected to be effective in suppressing or preventing cancer metastasis after cancer treatment (cancer resection surgery). Therefore, the present invention provides an agent for suppressing or preventing cancer metastasis, which comprises a green tea extract and a rosemary extract in a weight ratio of 10:1 to 1:10 (mixing weight ratio of green tea extract: rosemary extract) ( Hereinafter, the term "cancer metastasis inhibitor/preventive agent" is also included. In addition, the present invention requires the use of a green tea extract and a rosemary extract at a weight ratio of 10:1 to 1:10 (mixing weight ratio of green tea extract:rosemary extract) to suppress or prevent cancer metastasis. It also includes a method for suppressing or preventing cancer metastasis, which comprises administering to a patient.

Further, both the green tea extract and the rosemary extract used in the cancer cell migration inhibitor of the present invention are used in foods as antioxidants, spices and extracts having functionality, and are taken daily. It is present and safety is recognized. Therefore, the cancer cell migration inhibitor of the present invention is used as a health food or the like for the purpose of suppressing cancer cell migration, or for suppressing or preventing cancer metastasis after cancer treatment (cancer resection surgery). Or, it can be mixed with pet food and taken daily.

That is, according to the present invention, there is provided a safe cancer cell migration inhibitor (suppressant/prevention agent for cancer metastasis) that can substitute for the conventionally known suppression of cancer cell migration.

In the present specification, “cancer cell migration” or “cancer cell migration” means that a cancer cell moves from a place where the cancer cell has occurred (primary focus) to another position due to its motility. Types of cell migration include chemotaxis (Chemotaxis), in which cells migrate according to the concentration gradient of chemokine and other chemotactic factors, and the concentration of chemotactic factors in which cells bind to cell adhesion sites or extracellular matrix. Haptotaxis, which migrates according to the gradient, Wound Healing, in which cells migrate between wounds, and cells pass through the ECM through extracellular matrix (ECM) degradation and proteolysis. Cell invasion (Invasion) that migrates to adjacent tissues. Therefore, the cancer cell migration inhibitory effect can be evaluated using a known method, for example, inhibition of formation of a raffle membrane, a wound healing assay, or a commercially available cell migration measurement system. Here, the cancer cell migration inhibitory effect is evaluated using a wound healing assay.

The cancer cell migration inhibitor of the present invention contains a green tea extract and a rosemary extract in a weight ratio of 10:1 to 1:10 (mixed green tea extract:rosemary extract weight ratio). When the mixing ratio of the green tea extract and the rosemary extract is out of the above range, no significant difference in the cancer cell migration inhibitory effect is observed as compared with the green tea extract alone or the rosemary extract alone. Here, from the viewpoint of achieving a more remarkable synergistic effect of the cancer cell migration inhibitory effect, the cancer cell migration inhibitor is a green tea extract and a rosemary extract, preferably 5:1 to 1:5, more preferably 4:1 to 1:4 weight ratio, even more preferably 3:1 to 1:3 weight ratio, even more preferably 2:1 to 1:2 weight ratio, particularly preferably about 1:1 (substantially The same amount) (mixed weight ratio of green tea extract: rosemary extract). With the above mixing weight ratio, the effect of suppressing the migration of cancer cells by the combined use of the green tea extract and the rosemary extract can be particularly synergistically achieved.

Here, the target to be treated with the cancer cell migration inhibitor (patient in need of suppressing cancer cell migration or cancer metastasis) is not particularly limited, but is a mammal or a bird, preferably a mammal suffering from cancer or a bird. .. Here, mammals include primates such as humans, monkeys, gorillas, chimpanzees, orangutans, and non-human animals such as mice, rats, hamsters, guinea pigs, rabbits, dogs, cats, pigs, cows, horses, sheep, camels, and goats. Includes both human mammals. Examples of birds include chickens, quails, pigeons, and the like. Of these, humans, hamsters, guinea pigs, rabbits, dogs, cats, and pigs are preferable, and humans and pet animals such as dogs, cats, and rabbits are more preferable. That is, according to a preferred embodiment of the present invention, the cancer cell migration inhibitor of the present invention is orally used for humans or at least one pet animal selected from the group consisting of dogs, cats, and rabbits. According to a more preferred form of the present invention, the cancer cell migration inhibitor of the present invention is orally used in humans, dogs or cats. According to a particularly preferred embodiment of the present invention, the cancer cell migration inhibitor of the present invention is orally used in dogs or cats.

Since the cancer cell migration inhibitor of the present invention suppresses/inhibits cancer cell migration, it is expected to have an inhibitory/inhibitory effect also on cancer metastasis, which is a major obstacle to cancer treatment. That is, it is considered that the cancer cell migration inhibitor of the present invention is also effective in inhibiting and/or preventing cancer metastasis (herein, also simply referred to as “inhibition/prevention of cancer metastasis”). Therefore, the cancer cell migration inhibitor of the present invention is also useful as an agent for inhibiting/preventing cancer metastasis. As used herein, the term “metastasis of cancer” means that cancer is separated from the primary focus, migrates to another tissue or organ, and grows there. In addition, “suppression of cancer metastasis” or “suppression of cancer metastasis” refers to suppression of a phenomenon (metastasis) in which a cancer cell migrates (migrates) from the place where it originates (primary nest) and forms a tumor again at a remote site. Means that. Further, “prevention of cancer metastasis” or “prevention of cancer metastasis” means preventing or delaying the metastasis of cancer or tumor, or reducing the risk of metastasis of cancer or tumor. Therefore, in the present specification, the “cancer metastasis suppressive/preventive agent” means a drug exhibiting the suppressive/preventive effect of the above-mentioned cancer metastasis after cancer treatment (for example, surgery for removing cancer).

As described above, both the green tea extract and the rosemary extract used in the cancer cell migration inhibitor are used in foods as antioxidants, spices, and functional extracts, and can be taken daily. It is present and safety is recognized. Therefore, the cancer cell migration inhibitor of the present invention is ingested on a daily basis for the purpose of suppressing cancer cell migration, or for the purpose of preventing or suppressing cancer metastasis, as a health food, or mixed with pet food. It is also possible. That is, according to a preferred embodiment of the present invention, the cancer cell migration inhibitor of the present invention is a human health food or pet food composition.

Here, the type of cancer targeted by the cancer cell migration inhibitor (for suppressing/preventing cancer metastasis) is not particularly limited. For example, cancer of nervous system (eg, brain tumor, cervical cancer); cancer of digestive system (eg, oral cancer, pharyngeal cancer, esophageal cancer, gastric cancer, liver cancer, gallbladder cancer, biliary tract cancer, spleen cancer, colon cancer, small intestine cancer) , Duodenal cancer, colon cancer, colon adenocarcinoma, rectal cancer, pancreatic cancer, liver cancer); musculoskeletal cancer (eg, sarcoma, osteosarcoma, myeloma); urinary system cancer (eg, bladder cancer, renal cancer) ); genital cancer (eg, breast cancer, uterine cancer, ovarian cancer, testicular cancer, prostate cancer); respiratory cancer (eg lung cancer); hematopoietic cancer (eg acute or chronic myelogenous leukemia, Acute promyelocytic leukemia, leukemias such as acute or chronic lymphocytic leukemia, malignant lymphoma (lymphosarcoma), angiosarcoma, multiple myeloma, myelodysplastic syndrome, primary myelofibrosis, adventitial cell tumor); thyroid Cancer, parathyroid cancer, tongue cancer, malignant melanoma, mastocytoma, cutaneous histiocytoma, lipoma, hair follicle tumor, cutaneous papilloma, sebaceous adenoma, basal cell carcinoma and the like can be mentioned. Among these, the cancer cell migration inhibitor of the present invention is higher for hematopoietic cancer, digestive cancer, reproductive cancer, malignant melanoma (melanoma), mastocytoma, basal cell cancer Have an effect. That is, according to a preferred embodiment of the present invention, the cancer cell migration inhibitor of the present invention comprises hematopoietic cancer, digestive cancer, reproductive cancer, malignant melanoma (melanoma), mastocytoma and basal cell. It is used for suppressing and/or preventing metastasis of at least one disease selected from the group consisting of cancer. According to a more preferred embodiment of the present invention, the cancer cell migration inhibitor of the present invention is used for inhibiting and/or preventing metastasis of hematopoietic cancer or digestive cancer. According to a particularly preferred embodiment of the present invention, the cancer cell migration inhibitor of the present invention is used for suppressing and/or preventing metastasis of cancer of the digestive system.

(Green tea extract)
The green tea extract, which is an active ingredient of the cancer cell migration inhibitor of the present invention, can be obtained by extracting Camellia sinensis (all or part) using a suitable solvent. Here, the part of tea tree to be extracted is not particularly limited, and the whole plant body, leaves, stems, buds, flowers (including gaku, petals, pistil, stamen, etc.), wood part, bark part (bark), Fruits (including husks (flesh), ovary, pericarp (inner skin, mesocarp, outer skin) etc.), above-ground parts such as seeds; parts of plant bodies such as roots, rhizomes and underground parts such as tubers Include. The plant part may be subjected to extraction alone or in the form of a mixture of two or more kinds. Among these, considering the effect of suppressing/preventing migration of cancer cells, safety and the like, it is preferable to use tea leaves as a raw material. Alternatively, tea leaves (tea leaves) that have been heat-treated to prevent fermentation may be used for extraction. Specifically, the part of the tea tree that is the target of extraction is steamed and mashed raw leaves (tea leaves) of tea tree, and dried while adjusting the shape of the tea leaves (steaming ⇒ cooling ⇒ leafing ⇒ rough massage ⇒ massage ⇒ middle massage) ⇒Refining ⇒drying ⇒sieving and cutting ⇒separation of tree stems). In addition, in the present specification, a portion of the tea tree that is the target of extraction is also simply referred to as “green tea raw material”.

The green tea raw material may be subjected to extraction as it is, but may be dried and/or pulverized in advance before being subjected to extraction. Thereby, the desired active ingredient can be more efficiently extracted from the green tea raw material.

Here, the drying conditions for previously drying the green tea raw material are not particularly limited. Considering the crushability and the like, the drying temperature is preferably 20 to 90°C, more preferably 25 to 80°C. The drying time is preferably 24 to 120 hours, more preferably 48 to 96 hours. Alternatively, the green tea raw material may be dried by a freeze-drying powdering method, a high-pressure method, an ultra-high-pressure method, or the like. Among them, the high-pressure method is a method in which cells in the green tea raw material are pressure-crushed by performing high-pressure heat treatment at 100 to 150° C. for 2 to 10 hours (for example, 120° C. for 4 hours).

In addition, the crushing conditions for crushing the green tea raw material in advance are also not particularly limited. Considering extraction efficiency and the like, it is preferable that a predetermined plant part (green tea raw material) can be pulverized to a size of about 0.1 to 10 mm, more preferably about 0.5 to 5 mm. Examples of the crushing method include a method of shredding with a cutter, a slicer, a cutter, a peeler, a jaw crusher, a jet mill, a blender, and the like.

Next, if necessary, the previously dried and/or ground green tea raw material is extracted with a suitable solvent. The solvent that can be used here is not particularly limited as long as it can extract the active ingredient from the green tea raw material, and is appropriately selected depending on the plant or plant part to be used. Specifically, water (including tap water, industrial water, distilled water, reverse osmosis membrane water, filtered water, sterile water, purified water, etc.); methanol, ethanol, 1-propanol, 2-propanol, 1-butanol, Examples include alcohols such as 2-butanol, propylene glycol and 1,3-butylene glycol; esters such as methyl acetate and ethyl acetate; ketones such as acetone; ethyl ether, formaldehyde, dimethyl sulfoxide and the like. The above solvents may be used alone or in the form of a mixed solvent of two or more kinds. Of these, water, ethanol, or a mixed solution of water and ethanol is preferable as the extraction solvent in consideration of extraction efficiency and safety. That is, in a preferred embodiment of the present invention, the extract is an extract of the green tea raw material with water, ethanol or a mixed liquid of water and ethanol.

Moreover, the extraction may be performed only once, or may be performed twice or more. In the latter case, any solvent may be used in each extraction step, and the extraction conditions in each step may be the same or different. Extraction with water, hot water, ethanol or a mixed solution of water and ethanol is preferably performed at least twice, and more preferably extraction with water or a mixed solution of water and ethanol is performed after extraction with water or hot water. It is particularly preferable to carry out extraction with ethanol after extraction with hot water. By this method, the purity of the active ingredient (tea polyphenol which is the active ingredient) can be further improved.

If necessary, the green tea raw material may be extracted under acidic or alkaline conditions, that is, an acidic or alkaline solvent may be used. Here, the acid that can be used when preparing the acidic solvent is not particularly limited, but inorganic acids such as hydrochloric acid, sulfuric acid, nitric acid, phosphoric acid, acetic acid, citric acid, oxalic acid, succinic acid, formic acid, propionic acid, etc. Examples of organic acids include The alkali that can be used when preparing the alkaline solvent is not particularly limited, and examples thereof include potassium hydroxide, sodium hydroxide and sodium carbonate. The pH of the solvent used when extracting the green tea raw material under acidic or alkaline conditions is appropriately selected in consideration of the types of plants and plant parts to be used, extraction conditions, and the like. When an acidic solvent or an alkaline solvent is used, it is preferable to neutralize the extract after the extraction so that the extract becomes neutral (pH=about 7±1).

The amount of the solvent added is not particularly limited as long as the active ingredient can be extracted from the green tea raw material. Specifically, it is preferable to add the solvent in an amount of 10 to 1000 ml, more preferably 50 to 500 ml, based on 10 g of the green tea raw material.

The extraction conditions are also not particularly limited as long as the active ingredient can be extracted from the green tea raw material. Specifically, the extraction temperature is preferably 25 to 100°C, more preferably 40 to 100°C. The extraction time is preferably 30 minutes to 6 hours, more preferably 1 to 4 hours. Under such conditions, the active ingredient can be efficiently extracted from the green tea raw material.

After the extraction step, the solid matter (green tea raw material residue) is removed from the mixed solution of the green tea raw material and the solvent to separate the extract. Here, the separation method is not particularly limited, but examples thereof include filtration and centrifugation. Furthermore, this extract may be used as it is, but if necessary, it may be diluted with a diluent, extract by concentration, paste or solid form, frozen product form by freezing, dry powder product form by freeze-drying, etc. It may be converted into various forms (extracts). Here, the conversion method may be applied alone or in combination of two or more kinds. Preferably, the extract is concentrated to an appropriate concentration (for example, concentrated under reduced pressure), the obtained concentrate is frozen, and then freeze-dried, or the extract is made into a fine mist, which is then placed in hot air. A method of jetting and instantaneously obtaining a powdery dried product is preferably used.

The extract or extract may be further purified. Here, the purification method is not particularly limited, and a known purification method can be used. Specifically, a method in which a quaternary ammonium salt such as cetylpyridinium chloride is added to obtain a precipitate, and the precipitate is washed with a suitable solvent (eg, water or alcohol), anion exchange chromatography, cation Exchange chromatography, phosphocellulose chromatography, chromatography by hydrophobic reaction (hydrophobic interaction), affinity chromatography, chromatography using hydroxyapatite chromatography and lectin chromatography, recrystallization method, etc. To be

When caffeine is given to pet animals (eg, dogs), symptoms such as tachycardia, tremors/convulsions, arrhythmia, excessive excitement, and diarrhea may occur. Therefore, when the cancer cell migration inhibitor of the present invention is used in pet food (for example, for dogs), it is preferable that the cancer cell migration inhibitor does not contain caffeine as much as possible. Specifically, the content of caffeine in the green tea extract is preferably 5% by weight or less, more preferably 2.5% by weight or less, even more preferably 1% by weight or less, particularly preferably 0.5% by weight. % Or less (lower limit: 0% by weight). Therefore, caffeine may be reduced or removed before and after extraction from the green tea raw material as described above. Here, the method for reducing or removing caffeine is not particularly limited, and known methods can be used in the same manner or modified appropriately. Specifically, known methods such as JP-A-2014-140351, JP-A-2013-209428 and JP-A-2008-212024 can be mentioned.

Alternatively, the green tea extract may be a commercial product. Commercially available products include green tea extract powder (Matsuura Pharmaceutical Co., Ltd.), sun hood (registered trademark) powder, sun hood (registered trademark) 100, sun hood (registered trademark) CD, sun hood (registered trademark) aqueous solution, sun Sun Food (registered trademark) series such as Food (registered trademark) oil (all manufactured by Mitsubishi Kagaku Foods Co., Ltd.), green tea extract MF (manufactured by Maruzen Pharmaceutical Co., Ltd.), Sunphenon (registered trademark) 30S-OP, Sunphenon (registered trademark) Trademark) 30LB-OP, Sanphenon (registered trademark) EGCG-OP, Sanphenon (registered trademark) BG-3, Sanphenon (registered trademark) 90LB-OP, Sanphenon (registered trademark) 90S, Sunphenon (registered trademark) 90MB-OP, Sanphenon (Registered trademark) CF-T-OP, Sanphenon (registered trademark) NB-60, etc., Sanphenon (registered trademark) series (all manufactured by Taiyo Kagaku Co., Ltd.), polyphenon G, polyphenone (registered trademark) series such as 70A. , Sancatechin Aqueous F, Sancatechin Oil E, GTA-MNK-1 (all manufactured by Mitsui Norin Co., Ltd.), Tea Extract-40, Tea Extract-70, Tea Extract (decaffeinated) (all Tama Seikagaku Co., Ltd.) and the like.

The green tea extract may be used alone or in the form of a mixture of two or more kinds.

(Rosemary extract)
The rosemary extract, which is an active ingredient of the cancer cell migration inhibitor of the present invention, can be obtained by extracting rosemary (Rosmarinus officinalis L.) (all or part) with an appropriate solvent. Here, the portion of the rosemary to be extracted is not particularly limited, and the whole plant body, leaves, stems, buds, flowers (including rattle, petals, pistil, stamen, etc.), wood part, bark part (bark) , Fruits (including husks (flesh), ovary, pericarp (inner skin, mesocarp, outer pericarp), etc.), above-ground parts such as seeds; parts of plant bodies such as roots, rhizomes, underground parts such as tubers, etc. Includes. The plant part may be subjected to extraction alone or in the form of a mixture of two or more kinds. Of these, rosemary leaves are preferably used as a raw material in consideration of the effect of suppressing/preventing cancer cell migration, safety, and the like. In addition, in this specification, the part of the rosemary to be extracted is also simply referred to as “rosemary raw material”.

The rosemary raw material may be subjected to extraction as it is, but may be dried and/or pulverized in advance before being subjected to extraction. Thereby, the desired active ingredient can be more efficiently extracted from the rosemary raw material.

Here, the drying conditions for drying the rosemary raw material in advance are not particularly limited. Considering the crushability and the like, the drying temperature is preferably 20 to 90°C, more preferably 25 to 80°C. The drying time is preferably 24 to 120 hours, more preferably 48 to 96 hours. Alternatively, the rosemary raw material may be dried by a freeze-drying powdering method, a high-pressure method, an ultra-high-pressure method, or the like. Among these, the high-pressure method is a method in which cells in the rosemary raw material are pressure-crushed by high-pressure heat treatment at 100 to 150° C. for 2 to 10 hours (for example, 120° C. for 4 hours).

Further, the crushing conditions for crushing the rosemary raw material in advance are also not particularly limited. Considering extraction efficiency and the like, it is preferable that a predetermined plant part (rosemary raw material) can be pulverized to a size of about 0.1 to 10 mm, more preferably about 0.5 to 5 mm. Examples of the crushing method include a method of shredding with a cutter, a slicer, a cutter, a peeler, a jaw crusher, a jet mill, a blender, and the like.

Then, if necessary, the rosemary raw material which has been dried and/or ground in advance is extracted with a suitable solvent. The solvent that can be used here is not particularly limited as long as it can extract the active ingredient from the rosemary raw material, and is appropriately selected depending on the plant or plant part to be used. Specifically, water (including tap water, industrial water, distilled water, reverse osmosis membrane water, filtered water, sterile water, purified water, etc.); methanol, ethanol, 1-propanol, 2-propanol, 1-butanol, Examples include alcohols such as 2-butanol, propylene glycol and 1,3-butylene glycol; esters such as methyl acetate and ethyl acetate; ketones such as acetone; ethyl ether, formaldehyde, dimethyl sulfoxide and the like. The above solvents may be used alone or in the form of a mixed solvent of two or more kinds. Of these, water, ethanol, or a mixed solution of water and ethanol is preferable as the extraction solvent in consideration of extraction efficiency and safety. The lipophilic fraction contains a larger amount of the cancer cell migration inhibitory component. Therefore, from the viewpoint that the cancer cell migration inhibitory effect can be further improved, it is more preferable to extract from the rosemary raw material with ethanol or a mixed solution of water and ethanol. That is, in a preferred embodiment of the present invention, the extract is an extract of the rosemary raw material obtained by using ethanol or a mixed liquid of water and ethanol.

If necessary, the rosemary raw material may be extracted under acidic or alkaline conditions, that is, an acidic or alkaline solvent may be used. Here, the acid that can be used when preparing the acidic solvent is not particularly limited, but inorganic acids such as hydrochloric acid, sulfuric acid, nitric acid, phosphoric acid, acetic acid, citric acid, oxalic acid, succinic acid, formic acid, propionic acid, etc. Examples of organic acids include The alkali that can be used when preparing the alkaline solvent is not particularly limited, and examples thereof include potassium hydroxide, sodium hydroxide and sodium carbonate. The pH of the solvent when extracting the rosemary raw material under acidic or alkaline conditions is appropriately selected in consideration of the type of plant and plant part to be used, extraction conditions, and the like. When an acidic solvent or an alkaline solvent is used, it is preferable to neutralize the extract after the extraction so that the extract becomes neutral (pH=about 7±1).

The amount of the solvent added is not particularly limited as long as the active ingredient can be extracted from the rosemary raw material. Specifically, it is preferable to add the solvent in an amount of 10 to 1000 ml, more preferably 50 to 500 ml, based on 10 g of the rosemary raw material.

The extraction conditions are also not particularly limited as long as the active ingredient can be extracted from the rosemary raw material. Specifically, the extraction temperature is preferably 25 to 100°C, more preferably 40 to 100°C. The extraction time is preferably 30 minutes to 6 hours, more preferably 1 to 4 hours. Under such conditions, the active ingredient can be efficiently extracted from the rosemary raw material.

After the extraction step, the solid matter (residue of the rosemary raw material) is removed from the mixed solution of the rosemary raw material and the solvent, and the extract is separated. Here, the separation method is not particularly limited, but examples thereof include filtration and centrifugation. Furthermore, this extract may be used as it is, but if necessary, it may be diluted with a diluent, extract by concentration, paste or solid form, frozen product form by freezing, dry powder product form by freeze-drying, etc. It may be converted into various forms (extracts). Here, the conversion method may be applied alone or in combination of two or more kinds. Preferably, the extract is concentrated to an appropriate concentration (for example, concentrated under reduced pressure), the obtained concentrate is frozen, and then freeze-dried, or the extract is made into a fine mist, which is then placed in hot air. A method of jetting and instantaneously obtaining a powdery dried product is preferably used.

The extract or extract may be further purified. Here, the purification method is not particularly limited, and a known purification method can be used. Specifically, a method in which a quaternary ammonium salt such as cetylpyridinium chloride is added to obtain a precipitate, and the precipitate is washed with a suitable solvent (eg, water or alcohol), anion exchange chromatography, cation Exchange chromatography, phosphocellulose chromatography, chromatography by hydrophobic reaction (hydrophobic interaction), affinity chromatography, chromatography using hydroxyapatite chromatography and lectin chromatography, recrystallization method, etc. To be

Alternatively, the rosemary extract may be a commercial product. Commercially available RM keeper series such as rosemary extract (Matsuura Yakugyo Co., Ltd.), rosemary extract MF (Maruzen Pharmaceutical Co., Ltd.), RM keeper MP, RM keeper SF, RM keeper OS, RM keeper OSE (All manufactured by Mitsubishi Chemical Foods Co., Ltd.), RM-21 series such as RM-21A, RM-21S, RM-21A base, and RM-21B base (all manufactured by Mitsubishi Chemical Foods Co., Ltd.), Molka (rosemary extraction) Products, manufactured by Asama Kasei Co., Ltd.) and the like.

The rosemary extract may be used alone or in the form of a mixture of two or more kinds.

(Use of cancer cell migration inhibitor)
The cancer cell migration inhibitor of the present invention may be used for medical purposes (for the purpose of suppressing or preventing cancer metastasis) or may be used as a health food, except that it contains the cancer cell migration inhibitor of the present invention as an active ingredient. Can be used in a conventional dosage form. That is, the cancer cell migration inhibitor of the present invention is mixed with a pharmaceutically acceptable additive such as an excipient, and is suitable for parenteral administration, oral administration or external administration, pharmaceuticals, quasi drugs and food compositions. Can be used in the form of

Here, when the cancer cell migration inhibitor of the present invention is used in the form of a food composition, the cancer cell migration inhibitor is a fat or oil product, an emulsified product, a food such as a soft drink, and further a pet food or a health food. Can be added and used as. Here, the "food composition" means a composition intended to be ingested by mammals (including pets) such as humans. For example, a pet food composition is a food composition intended for ingestion by pets. Food compositions are widely known in the art. A pet food composition may or may not be nutritionally balanced and, in addition to supplements (eg, food), is a nutritionally balanced composition suitable for daily eating. May be. Here, "nutritionally balanced" means that the composition of the present invention has known amounts of nutrients necessary for sustaining life in an appropriate amount and ratio in the field of pet nutrition. To do.

In particular, when the cancer cell migration inhibitor of the present invention is added to and used in human health foods (human health foods) and pet foods, the addition amount (content) of the cancer cell migration inhibitor is not particularly limited. The amount is preferably 0.01 to 30% by weight, more preferably about 0.05 to 20% by weight, based on the health food and pet food (solid content conversion). Alternatively, the cancer cell migration inhibitor of the present invention is administered in an amount of preferably 0.1 to 1000 mg/kg body weight, more preferably 1 to 500 mg/kg body weight as a total weight per day. With such an amount, a sufficient cancer preventive effect can be achieved. In addition, if the amount is as described above, the pet takes the total amount of the given pet food without inhibiting the preference of the pet.

As described above, when caffeine is given to pet animals (for example, dogs), symptoms such as tachycardia, tremors/convulsions, arrhythmia, excessive agitation, and diarrhea may be caused. Therefore, when the cancer cell migration inhibitor of the present invention is used in pet food (for example, for dogs), it is preferable that the cancer cell migration inhibitor does not contain caffeine as much as possible. Specifically, the content of caffeine in the cancer cell migration inhibitor is preferably 10% by weight or less, more preferably 5% by weight or less, even more preferably 2% by weight or less, particularly preferably 1% by weight or less. (Lower limit: 0% by weight).

The pet food contains the same components as conventional except that it contains the cancer cell migration inhibitor of the present invention. For example, pet food, in addition to the cancer cell migration inhibitor of the present invention,
Monosaccharides, oligosaccharides, polysaccharides, dietary fibers, starches (for example, waxy corn starch, corn starch, wheat starch, rice starch, starch starch, potato starch, honey starch, tapioca starch, sago starch, chemically treated to these. Carbohydrate sources such as processed and chemically modified starch) and cereals (eg corn, barley, wheat, rye, sorghum, rice, loin, fluff, amarasanthus, quinoa); cattle, pigs, sheep, rabbits, kangaroos Meat and beef, etc., by-products and processed products thereof, chicken meat such as chicken, turkey, quail, etc., by-products and house products, fish meat such as fish and white fish, by-products and processed products, meat meal , Protein source such as soybean protein, wheat protein, wheat gluten, corn gluten, etc.; α-sitosterol; animal protein source such as lettering of the above raw materials such as meat bone, chicken meal, porridge meal, fish meal and the like; , Β-sitosterol, stigmasterol, campesterol, α-sitostanol, β-sitostanol, stigmasteranol, campestanol, cycloartenol, etc., free form thereof, fatty acid ester, ferulic acid ester, cinnamic acid ester, etc. Sterols such as ester form; rice bran, bran such as bran, meal such as soybean meal, vegetables such as vegetable extract, vitamins A, B1, B2, D, E, niacin, pantothenic acid, vitamins such as carotene And so on. In addition to the above, other additives generally used in pet foods such as gelling agents, shape-retaining agents, pH adjusters, seasonings, preservatives and nutritional supplements may be contained. In addition to or instead of the above-mentioned other additives, antioxidants such as butylhydroxyanisole (BHA), dibutylhydroxytoluene (BHT), ethoxyquin, tert-butylhydroquinone (TBHQ), propyl gallate, etc. It is also possible to use together. The addition amount (content) of each component described above is not particularly limited, and the same amount as the conventional amount can be applied.

The pet food is manufactured by a conventionally known method. For example, it can be produced by mixing the cancer cell migration inhibitor of the present invention and the above-mentioned necessary components to obtain a desired form. As an example, together with the cancer cell migration inhibitor of the present invention, grains, meat meal and mineral components such as iron and copper, if necessary, citric acid or citrate is mixed, and after sufficient mixing, water or steam. Extrude with an extruder while adding water. Then, it is preferably dried with hot air until the water content is 10% or less, to produce a pet food. When the pet food contains an oil or fat having a fatty acid having two or more double bonds, it is desirable that the pet food be dried with hot air and then coated.

The pet food may be in any form such as dry type, wet type, semi-moist type, jerky type, biscuit type, gum type, granular, powdery, soup type, etc., but the dry type is convenient for storage. It is preferable in terms of sex. Examples of dry pet foods include kibble shapes, flat plate shapes, and bone shapes. From the standpoint of obtaining a chewable and easy-to-handle shape for pets, the bulk density is 100 kg/m ³ or more, preferably 300 kg/m ³ or more, and 900 kg/m ³ or less, preferably 700 kg/m ³ or less. Yes, or 100 to 900 kg/m ³ , and particularly 300 to 700 kg/m ³ . Also, the pet food may be provided in bagged, boxed, packed, canned, retort pouched forms.

Further, the cancer cell migration inhibitor of the present invention can be used as a suppressant/preventive agent for cancer metastasis in medical applications. That is, the present invention provides a patient who needs to suppress cancer cell migration with a green tea extract and a rosemary extract at a weight ratio of 10:1 to 1:10 (green tea extract:rosemary extract mixed weight ratio). There is also provided a method of suppressing cancer cell migration, which comprises administering to the. In addition, the present invention suppresses cancer metastasis after excision of a green tea extract and a rosemary extract at a weight ratio of 10:1 to 1:10 (mixing weight ratio of green tea extract:rosemary extract) or Also provided is a method for suppressing or preventing cancer metastasis after cancer removal, which comprises administering to a patient in need of prevention. Here, the green tea extract and the rosemary extract may be administered in the form of a single agent containing both of these active ingredients, or may be administered simultaneously in the form of a dual agent containing these active ingredients separately. Good. Considering the synergistic effect of these active ingredients and the ease of administration, it is preferable to administer them in the form of a single agent containing both the green tea extract and the rosemary extract.

In this embodiment, the cancer cell migration inhibitor can be added to oral agents, external preparations, injections, inhalants, nasal drops/eye drops, etc., and tablets, solutions, injections, The desired dosage form can be an ointment, cream, lotion, aerosol, suppository, and the like. Further, if necessary, an excipient, a base, an emulsifier, a stabilizer, a solubilizing agent, a corrigent, a preservative, an aromatic, a coloring agent, a coating agent and the like can be appropriately mixed. As quasi-drugs/cosmetics, it can be added to lotions, emulsions, creams, etc., and if necessary, oils, moisturizers, UV absorbers, water-soluble polymers, antioxidants, surfactants, metals. Ion blocking agents, antibacterial preservatives, etc. can be added.

When the cancer cell migration inhibitor of the present invention is used as a drug, the dose is as follows: route of administration; patient's illness; patient size, weight, surface area, age and sex; other drugs administered; It depends on the judgment. A suitable dose (total amount of green tea extract and rosemary extract (dry weight conversion)) is 1 to 500 mg/kg body weight per day. Given the variety of available cancer cell migration inhibitors and the different efficacies of different routes of administration, it is expected that the required dosage can vary widely. Variations in these dosage levels can be adjusted using standard empirical procedures for optimality known in the art. Particularly for oral delivery, encapsulation of the cancer cell migration inhibitor in a suitable delivery vehicle (eg, polymer microparticles or implantable device) enhances delivery efficiency. In addition, the dose may be divided into one or more times a day. Alternatively, in some cases, it may be administered less frequently (eg, weekly or monthly). In addition, even in the same patient, the dose may vary depending on the patient's symptoms and severity.

When used as a drug (cancer cell migration inhibitor, cancer metastasis suppressor/preventive agent), a therapeutically effective amount of a cancer cell migration inhibitor is contained in one or more pharmaceutically acceptable carriers (additives) and And/or is formulated with a diluent. That is, the cancer cell migration inhibitor of the present invention may be provided in the form of a pharmaceutical composition further containing a pharmaceutically acceptable additive (eg, excipient, carrier, etc.). The cancer cell migration inhibitor of the present invention in this form can be specifically formulated for administration in solid or liquid form, as described in detail below. Examples of oral administration include drenches (aqueous or non-aqueous solutions or suspensions), tablets, boluses, powders, granules, and pastes for application to the tongue. For parenteral administration, for example, as a sterile solution or suspension, for example, for subcutaneous, intramuscular or intravenous injection, or for topical application, for example, as a cream, ointment or spray applied to the skin, or It can be formulated vaginally or rectally, for example as a vaginal suppository, cream or effervescent.

As used herein, "therapeutically effective amount", as used herein, is an effective benefit/risk ratio applicable to any medical treatment and is effective to produce any desired inhibitory effect. Amount of a different agent or cancer cell migration inhibitor. For example, although the dose of the cancer cell migration inhibitor of the present invention varies depending on the target disease, administration subject, administration route, etc., the cancer cell migration inhibitor of the present invention is orally administered for the purpose of suppressing/preventing cancer metastasis. In this case, the dose can be easily changed depending on the conditions such as the body weight of the target person, and thus can be appropriately selected by those skilled in the art. In the end, the attending physician will make an appropriate selection in consideration of the patient's symptoms and severity.

In the present specification, "pharmaceutically acceptable" means, within the scope of correct medical judgment, in proportion to a reasonable benefit/risk ratio, without problems such as excessive toxicity, irritation, allergic reaction or complication, Used to refer to a compound, material, composition, and/or dosage form suitable for use in contact with the tissue of the subject to be treated (human, mammal, etc.).

The pharmaceutically acceptable carrier means a liquid or solid filler or diluent that is involved in carrying or transporting the cancer cell migration inhibitor of the present invention from one organ or part of the body to another organ or part of the body. By pharmaceutically acceptable material, composition or excipient, such as an agent, excipient, solvent or encapsulating material. Each carrier must be “acceptable” in the sense of being compatible with the other ingredients of the dosage form and not injurious to the patient. Pharmaceutically acceptable carriers include, but are not limited to, sugars such as lactose, glucose and sucrose; starches such as corn starch and potato starch; celluloses such as sodium carboxymethyl cellulose, ethyl cellulose and cellulose acetate and its derivatives; Powdered tragacanth; malt; gelatin; talc; excipients such as cocoa butter and suppository waxes; oils such as peanut oil, cottonseed oil, safflower oil, sesame oil, olive oil, corn oil and soybean oil; glycols such as propylene glycol; Polyols such as glycerin, sorbitol, mannitol and polyethylene glycol; esters such as ethyl oleate and ethyl laurate; agar; buffers such as magnesium hydroxide and aluminum hydroxide; alginic acid; pyrogen-free water; isotonic saline. Ringer's solution; ethyl alcohol; phosphate buffered solutions; as well as other non-toxic compatible substances used in drug formulation. In some embodiments, the drug formulation is non-pyrogenic. That is, it is desirable that the body temperature of the patient is not increased. In addition, the wetting agents such as sodium lauryl sulfate and magnesium stearate, emulsifiers and lubricants, and coloring agents, releasing agents, coating agents, sweeteners, flavoring agents and flavoring agents, preservatives and antioxidants are cancer cells of the present invention. It may be included in a migration inhibitor (suppressor/preventer of cancer metastasis).

Pharmaceutically acceptable antioxidants include, but are not limited to, water-soluble antioxidants such as ascorbic acid, cysteine hydrochloride, sodium bisulfate, sodium disulfite, sodium sulfite, etc.; ascorbyl palmitate, butylhydroxyanisole ( BHA), butylhydroxytoluene (BHT), lecithin, propyl gallate, α-tocopherol, and other oil-soluble antioxidants; as well as citric acid, ethylenediaminetetraacetic acid (EDTA), sorbitol, tartaric acid, phosphoric acid, etc. A metal chelating agent is mentioned.

The cancer cell migration inhibitor of the present invention can be used in various dosage forms such as oral, nasal, topical (including buccal and sublingual), rectal, vaginal and/or parenteral administration. The dosage form may conveniently be presented in unit dosage form and may be prepared by any of the methods well known in the art of pharmacy. The amount of active ingredient that may be combined with the carrier materials to produce a single dosage form will vary depending upon the host treated, the particular mode of administration. The amount of active ingredient which can be combined with a carrier material to produce a single dosage form will generally be that amount of the compound which produces a therapeutic effect, but generally the amount of active ingredient (green tea extract and rosemary extract) will be The amount is about 0.1 to about 99% by weight, preferably about 1 to about 70% by weight, more preferably about 5 to about 50% by weight, based on the cancer cell migration inhibitor.

The methods of preparing these dosage forms or compositions include the step of bringing into association the cancer cell migration inhibitor of the invention with the carrier and, optionally, one or more accessory ingredients. In general, dosage forms are prepared by uniformly and intimately bringing into association the agent or agents of the invention with liquid carriers or finely divided solid carriers or both, and then, if necessary, shaping the product.

For example, dosage forms of the invention suitable for oral administration are in the form of capsules, sachets, pills, tablets, lozenges (using seasoned bases, usually sucrose and acacia or tragacanth), powders, granules. Or as a solution or suspension in an aqueous or non-aqueous liquid, or as an oil-in-water or water-in-oil liquid emulsion, or as an elixir or syrup, or pastilles (such as gelatin and glycerin, or sucrose and acacia). It may be used as an inert base) and/or a gargle and the like, each containing a predetermined amount of the compound of the present invention as an active ingredient. The agents of the present invention may be administered as a bolus, electuary or paste.

In the solid dosage forms of the invention for oral administration (capsules, tablets, pills, dragees, powders, granules, etc.), the active ingredients (green tea extract and rosemary extract) are sodium citrate or diphosphate diphosphate. Mixed with one or more pharmaceutically acceptable carriers such as calcium, and/or any of the following: fillers or extenders such as starch, lactose, sucrose, glucose, mannitol, and/or silicic acid. Binding agents such as carboxymethylcellulose, alginates, gelatin, polyvinylpyrrolidone, sucrose and/or gum arabic; moisturizers such as glycerol; agar, calcium carbonate, potato or tapioca starch, alginic acid, certain silicic acids. Salts and disintegrants such as sodium carbonate; dissolution retarders such as paraffin; absorption enhancers such as quaternary ammonium compounds; wetting agents such as cetyl alcohol and glycerol monostearate; such as kaolin and bentonite clay. Absorbents; lubricants such as talc, calcium stearate, magnesium stearate, solid polyethylene glycol, sodium lauryl sulphate, and mixtures thereof; and colorants. In the case of capsules, tablets and pills, the cancer cell migration inhibitor may comprise a buffer. Solid compositions of the same type can also be used as fillers in soft and hard filled gelatin capsules with excipients such as lactose or lactose and high molecular weight polyethylene glycols and the like.

Also, tablets may be made by compression or molding, optionally with one or more accessory ingredients. Compressed tablets include binders (eg gelatin or hydroxypropylmethylcellulose), lubricants, inert diluents, preservatives, disintegrants (eg sodium starch glycolate or crosslinked carboxymethylcellulose sodium), surface active agents. Alternatively, it may be prepared using a dispersant. Molded tablets may be made by molding in a suitable machine a mixture of the powdered compound moistened with an inert liquid diluent.

Solid dosage forms, such as tablets of the cancer cell migration inhibitor of the invention, such as sugar-coated tablets, capsules, pills and granules, are optionally scored or coated, such as enteric-coated coatings well known in the pharmaceutical dispensing art. And shells may be used. They have been used, for example, with hydroxypropylmethylcellulose, other polymeric matrices, liposomes and/or microspheres in various ratios to provide the desired release profile, to provide a sustained or controlled release of the active ingredient inside. It may be formulated to provide release. They may be sterilized, for example, by filtration through a bacteria-retaining filter, or by incorporating a sterilant in the form of a sterile solid composition which can be dissolved in sterile injectable medium such as sterile water immediately before use. .. These compositions may optionally contain an opacifying agent, only in certain parts of the gastrointestinal tract or preferentially there, optionally in a delayed manner, in a composition that releases one or more active ingredients. It may be. Examples of embedding compositions that can be used are polymeric substances and waxes. The active ingredient may also be in micro-encapsulated form, if appropriate, with one or more of the above-mentioned excipients.

Liquid dosage forms for oral administration of the cancer cell migration inhibitors of the present invention include pharmaceutically acceptable emulsions, microemulsions, solutions, suspensions, syrups and elixirs. Liquid dosage forms include, in addition to the active ingredient, inert diluents commonly used in the art, such as water and other solvents, ethyl alcohol, isopropyl alcohol, ethyl carbonate, ethyl acetate, benzyl alcohol, benzoic acid. Like benzyl, propylene glycol, 1,3-butadiene glycol, oils (especially cottonseed oil, peanut oil, corn oil, embryo oil, olive oil, castor oil and sesame oil), fatty acid esters of glycerol, tetrahydrofuryl alcohol, polyethylene glycol and sorbitan. Solubilizers and emulsifiers, and mixtures thereof. In addition to inert diluents, oral compositions may also include adjuvants such as wetting, emulsifying and suspending agents, sweetening agents, flavoring agents, coloring agents, flavoring agents and preservatives. The suspension may contain, in addition to the active compound, a suspension such as, for example, ethoxylated isostearyl alcohol, polyoxyethylene sorbitol and sorbitan esters, microcrystalline cellulose, aluminum metahydroxide, bentonite, agar and tragacanth, and mixtures thereof. A turbidity agent may be included.

The dosage form of the cancer cell migration inhibitor of the invention for rectal or vaginal administration may be presented as a suppository. This suppository is prepared by admixing one or more agents of the invention with one or more suitable non-irritating excipients or carriers including, for example, cocoa butter, polyethylene glycol, suppository waxes or salicylates. , Which is a solid at room temperature, but is a liquid at body temperature, will melt in the rectal or vaginal cavity and release the active compound. Dosage forms suitable for vaginal administration also include pessaries, tampons, creams, gels, pastes, effervescent or spray formulations containing such carriers as are known in the art to be appropriate.

Dosage forms for topical or transdermal administration of one or more cancer cell migration inhibitors of the present invention include powders, sprays, ointments, pastes, creams, lotions, gels, solutions, patches and inhalants. The active ingredients (green tea extract and rosemary extract) may be mixed under sterile conditions with a pharmaceutically acceptable base material and with any preservatives, buffers or propellants which may be required. Ointments, pastes, creams and gels, in addition to the active ingredients (green tea extract and rosemary extract), animal or vegetable fats, oils, waxes, paraffins, starches, tragacanth, cellulose derivatives, polyethylene glycols, silicones, bentonites. , Silicic acid, talc and zinc oxide, or mixtures thereof.

Powders and sprays contain, in addition to the active ingredients (green tea extract and rosemary extract), complementary forms such as lactose, talc, silicic acid, aluminum hydroxide, calcium silicates and polyamide powder, or mixtures of these substances. You may include a medicine. Sprays may additionally contain customary propellants such as chlorofluorohydrocarbons and volatile unsubstituted hydrocarbons such as butane and propane.

Transdermal patches have the added advantage of providing controlled delivery of a cancer cell migration inhibitor of the present invention to the body. Such dosage forms can be made by dissolving or dispersing the cancer cell migration inhibitor of the present invention in a suitable medium. Absorption enhancers can also be used to increase the flux of a substance that contains the cancer cell migration inhibitor of the present invention across the skin. The rate of such flux can be controlled by either providing a rate controlling membrane or by dispersing the compound in a polymer matrix or gel.

The cancer cell migration inhibitor of the present invention suitable for parenteral administration comprises one or more pharmaceutically acceptable sterile isotonic aqueous or non-aqueous solutions, dispersions, suspensions or emulsions, or immediately before use, together with the composition. Include sterile powders that can be reconstituted in sterile injectable solutions or dispersions, which include antioxidants, buffers, bacteriostats, solutes or suspensions that make the preparation isotonic with the blood of the intended recipient. Agents or concentrates may be included.

Examples of suitable aqueous and non-aqueous carriers that can be used in the cancer cell migration inhibitor of the present invention include water, ethanol, polyols (eg, glycerol, propylene glycol, polyethylene glycol, etc.), and suitable mixtures thereof, olive oil. There are vegetable oils such as, as well as injectable organic esters such as ethyl oleate. The inherent fluidity can be maintained, for example, by the use of a coating material such as lecithin, by the maintenance of the required particle size in the case of dispersion and by the use of surfactants.

The cancer cell migration inhibitor of the present invention may include auxiliary agents such as preservatives, wetting agents, emulsifying agents and dispersing agents. Prevention of microbial activity can be ensured by the inclusion of various antibacterial and antifungal agents such as paraben, chlorobutanol, phenol sorbate and the like. It may be desirable to include isotonic agents, such as sugars, sodium chloride, and the like into the compositions. In addition, sustained absorption of the injectable drug form may be brought about by the inclusion of agents which delay absorption such as aluminum monostearate and gelatin.

The cancer cell migration inhibitor of the present invention can also be used as a food (food composition). At this time, the cancer cell migration inhibitor of the present invention may be used as it is, liquid, gel-like or solid food, such as juice, soft drink, tea, soup, soy milk, salad oil, dressing, yogurt, jelly, pudding, Sprinkle, powdered baby milk, cake mix, powdered or liquid dairy products, bread, added to cookies, etc., if necessary, excipients and flavors such as dextrin, lactose, starch, and pellets, tablets, etc. It can be used as a health food, a dietary supplement, etc. by processing it into granules or the like, or by coating it with gelatin etc. to form a capsule. The blending amount of the cancer cell migration inhibitor of the present invention in these foods or edible compositions is difficult to uniformly define depending on the type and state of the food or composition, but is 0 relative to the total weight of the food. 0.01 to 90% by weight, more preferably 0.1 to 80% by weight. With such a blending amount, the effect of the cancer cell migration inhibitor can be sufficiently exerted without impairing the flavor. Also, the food can be easily prepared.

Furthermore, the cancer cell migration inhibitor of the present invention can be used as a cosmetic (cosmetic composition). At this time, examples of the form of the cosmetic (cosmetic composition) include lotions, emulsions, creams, and powders, but are not particularly limited thereto. Such a cosmetic product (cosmetic composition) containing the cancer cell migration inhibitor of the present invention can be produced using a method known to those skilled in the art. When the cancer cell migration inhibitor of the present invention is used as a cosmetic (cosmetic composition), the form includes, but is not particularly limited to, lotion, emulsion, cream, powder and the like. Such a cosmetic composition containing the cancer cell migration inhibitor of the present invention can be produced using a method known to those skilled in the art.

The effects of the present invention will be described using the following examples and comparative examples. However, the technical scope of the present invention is not limited to the following examples. In the following examples, the operations were performed at room temperature (25°C) unless otherwise specified. Further, unless otherwise specified, “%” and “part” mean “wt %” and “part by weight”, respectively.

Example 1 and Comparative Examples 1 to 3: Evaluation of Cell Migration Inhibition on Colorectal Cancer Cells The effect of the cancer cell migration inhibitor of the present invention on colon cancer cell migration inhibition was evaluated by the following wound healing assay.

(Sample preparation)
Rosemary extract (Mitsubishi Chemical Foods Co., Ltd., RM-21B base) and green tea extract (Mitsui Norin Co., Ltd., Polyphenon (registered trademark) 70A, caffeine content: 0.5 wt% or less), rose It was prepared with dimethyl sulfoxide (DMSO) so that the weight ratio of the mary extract and the green tea extract (the mixing weight ratio of the rosemary extract:green tea extract) was 1:1 (Example 1: Sample 1). Concentration (total concentration) of the mixture of rosemary extract and green tea extract: 40 mg/ml). In addition, as comparison targets, a rosemary extract (Comparative Example 1: Sample 2) and a green tea extract (Comparative Example 2: Sample 3) were each prepared in DMSO at 40 mg/ml.

(Wound healing assay)
Human colon adenocarcinoma-derived DLD-1 cells were cultured in RPMI1640 medium containing 10 (v/v)% FBS at 37° C. in a 5 vol% CO ₂ atmosphere. 2 ml of this cultured cell was seeded on a 6-well plate so that 5×10 ⁴ cells were formed per well. After culturing the DLD-1 cells at 37° C. for 72 hours in a 5 vol% CO ₂ atmosphere, the cells were scraped off at regular intervals (width: about 240 mm) using a micropipette tip (FIG. 1 below). See the photo on the left). The medium was replaced with a fresh RPMI1640 medium containing 10 (v/v)% FBS, and 2 μl of each of Samples 1 to 3 prepared above was added to the medium so that the final concentration was 40 μg/ml. As a control, only 2 μl of the solvent (DMSO) was added. Immediately after the addition of each sample or DMSO (after 0 hour) and after the above addition and after culturing at 37° C. for 10 hours in a 5% CO ₂ atmosphere, the appearance of DLD-1 cells is photographed, and the cell migration ability is shown. An evaluation was made. Four areas were randomly selected from each photograph, and the area where cells migrated was measured using image processing software GNU Image Manipulation Program, and the average value and standard deviation thereof were calculated. The results are shown in FIGS. 1 and 2. In FIG. 1, FIG. 1(a) shows a 1:1 mixture of rosemary extract and green tea extract (sample 1), FIG. 1(b) shows a rosemary extract (sample 2), and FIG. c) shows the green tea extract (Sample 3), and FIG. 1(d) shows the control (DMSO only). In addition, in FIG. 2, each value represents the migration area of DLD-1 cells to which each sample was added, where the migration area of the control was 100%, as an average value±standard deviation (n=4). .. In FIG. 2, “*” indicates that there is a particularly significant difference (p<0.01) when compared with the rosemary extract alone and the green tea extract alone.

As is clear from FIGS. 1 and 2, sample 1 (Example 1; 1:1 mixture of rosemary extract and green tea extract; FIG. 1(a), second from the left in FIG. 2) was sample 2 ( Comparative Example 1; rosemary extract alone; FIG. 1(b), FIG. 2 left third) and sample 3 (Comparative Example 2: green tea extract alone; FIG. 1(c), FIG. 2 left fourth), It can be seen that the colon cancer cell migration inhibitory effect is significantly higher than that of the control (no addition; FIG. 1(d), the first from the left in FIG. 2). From FIGS. 1 and 2, by using the cancer cell migration inhibitor of the present invention, the cancer cell migration inhibitory effect can be synergistically improved as compared with the rosemary extract alone and the green tea extract alone. Is considered.

From the above results, it is considered that the cancer cell migration inhibitor of the present invention can be used for inhibiting/preventing cancer metastasis of digestive system such as colon cancer. Further, both the green tea extract and the rosemary extract that constitute the cancer cell migration inhibitor of the present invention are used in foods as antioxidants, spices, and functional extracts, and are recognized as safe. There is. Therefore, the food or pet food containing the composition of the present invention is expected to be useful as a health food for suppressing/preventing cancer metastasis of digestive system such as colon cancer.

In this example, the cancer cell growth inhibitory effect on human cells was evaluated, but it is speculated that similar results can be obtained on pet animals such as dogs and cats.

Claims (5)

It contains only green tea extract and rosemary extract as an active ingredient, and the green tea extract and rosemary extract are in a weight ratio of 2:1 to 1:2 (mixing weight ratio of green tea extract: rosemary extract). A colon adenocarcinoma cell migration inhibitor. The colon adenocarcinoma cell migration inhibitor according to claim 1, comprising a green tea extract and a rosemary extract in a weight ratio of about 1:1 (mixed weight ratio of the green tea extract:rosemary extract). The colon adenocarcinoma cell migration inhibitor according to claim 1 or 2, which is used for suppressing and/or preventing metastasis of colon adenocarcinoma . Human or dog, is used at least one pet animals orally is selected from the group consisting of cats and rabbits, colon adenocarcinoma cell migration inhibitor according to any one of claims 1-3. Is a health food or pet food composition for human, colon adenocarcinoma cell migration inhibitor according to any one of claims 1-4.