WO2007072965A1

WO2007072965A1 - Angiogenesis inhibitor and utilization of the same

Info

Publication number: WO2007072965A1
Application number: PCT/JP2006/325687
Authority: WO
Inventors: Koji Suzuki; Joji Otani; Shigeo Tanaka
Original assignee: Konan Chemical Manufacturing Co., Ltd.; Mie University
Priority date: 2005-12-21
Filing date: 2006-12-19
Publication date: 2007-06-28

Abstract

It is intended to provide a novel angiogenesis inhibitor. For achieve thus purpose, an angiogenesis inhibitor which contains a sulfated polysaccharide having rhamnose as the main constituting monosaccharide or its salt is provided.

Description

TECHNICAL FIELD Field of the Invention

[00 0 1] The present invention relates to an angiogenesis inhibitor and use thereof, and more specifically, an angiogenesis inhibitor containing a sulfated polysaccharide containing rhamnose as a main component of a constituent monosaccharide and an angiogenesis comprising the inhibitor. The present invention relates to a pharmaceutical composition or the like for preventing or treating a disease requiring inhibition. '

Background art

[000 2] In vivo, angiogenesis occurs under various physiological conditions, but during cancer growth and metastasis, cancer cells induce vascular endothelial growth factor (VEGF) for their own nutrition. It is thought to cause angiogenesis. For this reason, attention has been focused on methods for inhibiting tumor cell angiogenesis to suppress the growth and metastasis of cancer cells, and agents that inhibit angiogenesis have also been disclosed (Japanese Patent Application Laid-Open No. 2000-3-9). 6 0 7 8).

[00 0 3] On the other hand, as an algae-derived acidic polysaccharide, fucodyne, rhamnan sulfate, galactan sulfate and the like are known. For example, fucodyne, which is a sulfated polysaccharide, has anticancer activity and anticoagulant activity. .Activity is known. Similarly, rhamnan sulfate, which is a sulfated polysaccharide, is known to have anticoagulant activity (Japanese Patent Application Laid-Open No. 6 3-2 3 5 3 0 1). Disclosure of the invention

[00 04] However, the effect of rhamnan sulfate on angiogenesis has not been reported. Accordingly, an object of the present invention is to provide a novel angiogenesis inhibitor capable of inhibiting angiogenesis, a pharmaceutical composition, a food composition, and the like using the inhibitor.

[000 5] The inventors of the present invention have found that the mosquitoes belonging to the green alga plant, Motonae (M onostromanitidum), ヒロノヽノヒトグサ (M onostroma L atissimum) was found to show an angiogenesis inhibitory action, and the present invention was completed. That is, according to the present invention, the following means are provided.

[0 00 6] According to one aspect of the present invention, there is provided an angiogenesis inhibitor comprising a sulfated polysaccharide containing rhamnose as a main component of a constituent monosaccharide or a salt thereof. In this form, the sulfated polysaccharide can have the following characteristics.

(a) In the total mass of the constituent polysaccharide, the rhamnose content is 5.0% by mass or more, the -dulcose content is 1% by mass to 30% by mass, and the xylose content is 1 mass. /. 10 mass% or less

(b) The sulfate group content is 1.0 mass% or more and 40 mass% or less

[0 00 7] Furthermore, the angiogenesis inhibitor of the present invention may have a pullulan-converted weight average molecular weight of 50,000 to 300,000 by gel filtration chromatography.

[0 0 0 8] In the present embodiment, the sulfated polysaccharide can be derived from algae belonging to the moss and moss algae class. The inhibitor may include the algae or the algae extract, and the extract may be a hot water extract of the algae. Furthermore, the algae can be selected from the power of algae belonging to the genus Mon o s tr o m a, and can use M o n o s t r. O ma La t i s s i m u m or M o n s t r o mani d i dum. .

[0 00 9] According to another embodiment of the present invention, there is provided an angiogenesis inhibitor containing an algae belonging to the genus Monostroma or an extract thereof. In this embodiment, the inhibitor preferably contains a sulfated polysaccharide having rhamnose as a main component of a constituent monosaccharide or a salt thereof.

[0 0 10] According to another embodiment of the present invention, it is also a preferred embodiment that the sulfated polysaccharide is a sodium salt.

[0 0 1 1]

According to another aspect of the present invention, there is provided a pharmaceutical composition for preventing or treating a disease requiring angiogenesis inhibition, comprising any one of the above angiogenesis inhibitors. Provided. In this form, the disease can be selected from the group consisting of solid tumors, vascular lesions, skin diseases, osteoarthritis, genital diseases, eye diseases and lung diseases. Further, the disease may be a solid tumor metastasis. In addition, the pharmaceutical composition can be used for topical administration or can be used for oral administration. '.:

[00 1 2] According to another embodiment of the present invention, there is provided a food composition for preventing or prognosing a disease requiring angiogenesis inhibition, comprising any one of the above-mentioned angiogenesis inhibitors. . The food composition may be a nutritional supplement composition. Furthermore, the food composition can be used for inhibiting metastasis of solid tumors. Brief description of the drawings ''

[00 1 3] Figure 1 shows the sample:! FIG. 5 is a graph showing the lumen formation rate (%) of ˜3.

[00 1 4] FIG. 2 is a graph showing the lumen formation rate (%) of Samples 4-8. '

[00 15] FIG. 3 is a diagram showing lung metastasis of B 16 mouse melanoma cells.

[00 16] FIG. 4 is a diagram showing the lung metastasis rate of B 16 mouse melanoma cells. BEST MODE FOR CARRYING OUT THE INVENTION

[0 0 17] The angiogenesis inhibitor of the present invention is characterized by containing a sulfated polysaccharide containing rhamnose as a main component of a constituent monosaccharide or a salt thereof. By administering the angiogenesis inhibitor of the present invention, it becomes possible to prevent or treat diseases that require angiogenesis inhibition. Furthermore, by taking this inhibitor, a patient with a disease requiring inhibition of angiogenesis or an individual who needs a risk for the disease, specifically biochemically, physically or genetically, has an indication of the disease. In an individual, an individual after treatment of the disease, or an individual who is predicted to develop such a disease as a derivative disease, the onset of the disease is prevented or delayed, the progression is suppressed, and the prognosis is reduced. Can improve and improve QOL.

[0 0 1 8] In this specification, “angiogenesis” means that a new blood vessel is formed from an existing blood tube. Therefore, angiogenesis, which means the first angiogenesis process observed in the early stage of embryogenesis, is not included in the term “angiogenesis” in the present invention. In the present specification, angiogenesis refers to angiogenesis of animals including mammals and non-mammals. Mammals include humans and non-human animals, and non-human animals include domestic animals such as sushi, pigs, hidges, goats and hooters, chickens and turkeys. Etc. Includes pets such as poultry, dogs and cats.

[0 0 19] Hereinafter, the angiogenesis inhibitor, the pharmaceutical composition, the food composition and the like of the present invention will be described in detail. · '■

(Angiogenesis inhibitor) The angiogenesis inhibitor of the present invention (hereinafter simply referred to as the present inhibitor) contains a sulfated polysaccharide containing rhamnose as a main component of a constituent monosaccharide or a salt thereof. Examples of the sulfated polysaccharide in the present invention include a sulfated polysaccharide in which the 3-position or 4-position of the constituent sugar is sulfated. The sulfated polysaccharide may be a free acid in its sulfate group or a salt such as a metal salt. Examples of such salts include alkali metal salts such as sodium and potassium, and alkaline earth metals such as calcium and magnesium. In the present description, a sodium salt is preferable.

[0 0 2 0] The monosaccharide composition of the sulfated polysaccharide in the present invention is mainly composed of rhamnose and 50 masses of the constituent monosaccharide. /. The above is rhamnose. The rhamnose molar ratio in the sulfated polysaccharide is preferably 60% by mass or more, more preferably 70% by mass or more. The relationship between the rhamnose ratio and the angiogenesis inhibitory activity is not necessarily clear and does not restrict the present invention.

[0 0 2 1] Regarding the composition of the constituent monosaccharides in the sulfated polysaccharide, for example, after the polysaccharide is decomposed to the constituent monosaccharide by the acid hydrolysis method, the free acid is separated and removed, and the monosaccharide components are separated by HPLC. It can be obtained by quantification. The acid hydrolysis method can be performed, for example, as follows. The saccharide-containing sample was dissolved in acid hydrolysis <polysaccharide sample (approximately 20 mg) in 2 N sulfuric acid (10 ml), After heating in a boiling water bath for 2 hours>, 10% aqueous solution of barium chloride is added, and free sulfuric acid is precipitated as barium sulfate (centrifugation), and the supernatant is collected in a high-pressure liquid chromatography. (Column: A sahipack NH 2 P—500 4 E, column temperature: 35 ° C, mobile phase: water acetonitrile = 25_75, flow rate: 1 ml / min, detector: RI) .

[0 0 2 2] The sulfated polysaccharide of the present invention may contain glucose and Z or xylose as a constituent monosaccharide other than rhamnose. Preferably, it contains glucose, and further contains xylose. If it contains glucos, its content is 1 mass. / _{0 or} more 30 It is preferable that it is 0 mass% or less. When xylose is contained, the content is preferably 1% by mass or more and 10% by mass or less. These constituent monosaccharides can also be calculated in the same manner as the ramnose content.

[0 0 2 3] The molecular weight of the sulfated polysaccharide of the present invention by gel chromatography (GPC) is not particularly limited as long as it has angiogenesis inhibitory activity. For example, a pullulan-converted weight average molecular weight of 50,000 to 300,000 can be used. Considering that this inhibitor or composition is administered orally, for example, those having a weight-average molecular weight in pullulan conversion of 250,000 or less can be preferably used. On the other hand, in the case of local administration by injection or the like, for example, those having a weight average molecular weight in terms of bullan of more than 300,000 or less and less than 300,000 can be preferably used.

[0 24] The sulfated polysaccharide of the present invention may be one obtained by appropriately hydrolyzing such a naturally-derived sulfated polysaccharide as necessary. In addition, as the sulfated polysaccharide of the present invention, various molecular weight fractions thus fractionated by GPC or the like can be used alone or in combination of two or more kinds. In addition, as a fractionation method based on molecular weight, for example, in addition to GPC, an ultrafiltration method can be used alone or in combination with other molecular weight fractionation methods. Fractions separated by these various fractionation methods may be combined.

[0 0 25] The sulfate group content in the sulfated polysaccharide of the present invention is not particularly limited as long as it has an angiogenesis inhibitory activity, but is generally 15 masses. / ₀ or more And are preferred. The upper limit is not particularly limited, but is preferably 40% by mass or less. More preferably, it is 35 mass% or less. The sulfuric acid and group content can be determined by the combustion flask method and the rosin acid method. Combustion Flasco methods, for example, Analytical Chemistry, 15 5, 6 8 9-6 9 1, (1 9 6 6), 1 7, 1 3 2 2-1 3 2 4, (1 9 6 8) can be used. By quantifying the sulfur element in the sample by the combustion flask method and multiplying the sulfur atom content (% by mass) in the obtained sample by 3, the sulfate group content can be calculated. In addition, the mouth-Dison method is the same as the method described in Example-Eiichi (Ma, A na 1 ytica 1 B idchem, 4 1 卷, 4 7:! ~ 4 7 6, (1 9 7 1) Can be used.

[0 0 26] Such sulfated polysaccharides include, in nature, acidic polysaccharides that are contained in algae belonging to the class of alga alga and extracted from the algae. Therefore, the inhibitor can contain an algae belonging to the class of alga algae containing such sulfated polysaccharide, an extract from the algae, or a part thereof. Although not restricting the present invention, the sulfated polysaccharide of the present invention is included in the inhibitor together with the extract of the alga that is the plant from which the sulfated polysaccharide is contained. It is considered preferable in view of the physiological activity of the sulfated polysaccharide. Therefore, as the present inhibitor, such algae or an extract thereof can be preferably used. More preferably, it contains the sulfated polysaccharide in the present invention, and more preferably, the extract fraction containing the sulfated polysaccharide of the present invention is used.

[0 0 2 7] In order to obtain a sulfated polysaccharide from algae, for example, the following method can be employed. The dried algae are washed and swollen with water, and then extracted with an aqueous medium such as water. As the aqueous medium, water is used as a main component, and preferably, only water or an acidic aqueous medium containing water and an organic acid and / or an inorganic acid is used. Examples of acids that can be used include succinic acid, succinic acid, malic acid, fumaric acid, oxalic acid, L-ascorbic acid, acetic acid, hydrochloric acid, sulfuric acid and the like alone or in combination. Preferably, the raw material is appropriately shredded during extraction. Extracting sulfated polysaccharides with aqueous media can be done by heating the aqueous media. It is preferably carried out with hot water (70 ° C or higher, preferably about 95 ° C or higher). The extraction time is approximately 1 to 8 hours. After extraction, the precipitate is removed by centrifugation, and then the supernatant is dialyzed as it is or an organic solvent such as ethanol is added to precipitate sulfated polysaccharide, and then the aqueous 5 solution of the precipitate is dialyzed. By removing low molecular components, a crude sulfated polysaccharide fraction can be obtained. This crude fraction can be used as the inhibitor of the present invention as it is, but this crude fraction can be further purified by applying an ionic strength gradient using a DEAE cellulose column or the like. -In addition, in such a sampling-collection operation, bioassay is performed in a timely manner to inhibit the formation of the lumen.

It is preferable to proceed with purification using 10 as an index.

[0 0 2 8] Also, algae-derived sulfated polysaccharides such as rhamnan sulfate are mainly in the form of Na, K, 'Ca, Mg. Salt, etc., but the sulfated polysaccharide of the present invention. Can take various forms without being limited to the natural salt form. To change the type of salt, the sulfate groups of sulfated polysaccharides can be exchanged with ions.

From No. 15, it should be free and then neutralized with the desired alkali. For example,

'The sodium salt of a sulfated polysaccharide in the present invention can be obtained as follows. That is, sulfate groups can be made free by using a cation exchange resin column or the like in which sulfated polysaccharides are equilibrated with hydrochloric acid or the like, and then sulfate sulfate-free polysaccharides can be neutralized with NaOH. .

20 [0 0 2 9] Examples of these algae include the powers of the eyes of Mongosto tromo, P ro te o omno s t ro ma, and the moss B 1 i n d i n g i a, En t e r o m o r p h a, U l v a, U 1 v a r i a. Specifically, Mon o s t r om a n i t i d um (Hytogusa), M o n o s t r o m a 1 a t i s s i m u m (Hirono ヽ no hi toku "sa), Mo n o

25 str om a S ravillei, Mo nostromaang ι cava 1 is (Buanonori), Enteromorphacom Examples include pressa, Enteromorpha 1 inza, U lvapertusa, U 1 variafusca, B 1 indingiaminima. Of these, hitohedasa, hirohanohi todasa and the like are preferably used. Hitoedasa and Hirohano Hitoedasa are similar, and it is not always easy to distinguish them. Therefore, unless the algae can be clearly classified as either of them, the algae can be treated as Hitoedasa or Hiroha no Toedasa. --, [0 0 3 0] Examples of sulfated polysaccharides include naturally occurring polysaccharides or semi-synthetic sulfated polysaccharides chemically modified from sulfated polysaccharides as long as they have angiogenesis inhibitory activity. . For example, sulfated polysaccharides or their degradation products collected from natural organisms such as the algae described above may be sulfated, or partially desulfated. . Furthermore, a naturally occurring polysaccharide that does not have a sulfate ester group or a degradation product thereof may be sulfated. Sulfation can be achieved by sulfonating the hydroxyl group). Sulfonation of the hydroxyl group can be achieved by, for example, chlorosulfonic acid monopyridine complex, DCC (dicyclohexyl carbodiimide) monosulfate, trioxide trimethylamine complex, N, N-dimethylformamide (DMF) trioxide It can be carried out by a method using a pyridine complex.

[0 0 3 1] Furthermore, as a sulfated polysaccharide, as long as it has angiogenesis inhibitory activity, a synthetic sulfated polysaccharide such as a sulfated synthetic polysaccharide or a sulfated monosaccharide or oligomerized polymer is used. Also mentioned. Furthermore, the sulfated polysaccharide of the present invention may be a sulfated polysaccharide that has been subjected to a general chemical modification applied to a sugar with respect to a constituent monosaccharide other than rhamnose as long as it has angiogenesis inhibitory activity. Good. Examples of the chemical modification include acylation of hydroxyl group and the like, acetylation, force alkoxylation, phosphorylation and the like. Those skilled in the art can carry out chemical modification methods for such sugars as needed.

[0 0 3 2] The present inhibitor is a combination of one or more of these sulfated polysaccharides or an algae containing such sulfated polysaccharides or an extract thereof. Can be used. The angiogenesis inhibitory activity of this inhibitor can be evaluated by, for example, a generally used method using lumen formation as an index. An example of the evaluation of angiogenesis inhibition using lumen formation as an index will be described below. First, cell suspensions of human umbilical cord-derived venous endothelial cells (HUVE C) with a predetermined cell concentration are prepared, sulfated polysaccharides of various concentrations are added to this suspension, and this mixture is added to Matrigel (trademark). Seed on a gel for cell culture such as culturing in a CO 2 incubator at 37 ° C for 6 hours, and then observing with a phase contrast microscope. The length of the lumen formed by the HUVEC is measured with an image analyzer after obtaining this image information with a camera. The length of the lumen is calculated, for example, by measuring the length of the lumen formed in at least four fields of view selected from images taken using a phase contrast microscope equipped with a CCD camera, and calculating the average value. Can do. The inhibition rate is the length of the lumen formed in the absence of the sulfated polysaccharide (mean value) The length of the lumen in the presence of monosulfated polysaccharide (mean value) The length of the lumen in the absence of sulfated polysaccharide X 1 It can be calculated as 00 (%). , '

[0 0 3 3] In addition, for this inhibitor, for example, angiogenesis' inhibitory activity ((1) and (2)) and cancer cell growth / metastasis-suppressing effect (3) can be confirmed by the following method.

[00 34] (1) Angiogenesis inhibitory activity—Matri.Gel (trademark) Implant Atsusei Sulfated polysaccharide-containing samples such as extract of edible fish at various concentrations in the presence or absence of VEGF and heparin It is mixed with Matrigel below, and this Matrigel is injected subcutaneously into severe combined immunodeficiency (SCID) mice. Three days later, the amount of blood vessels newly born toward Matrigel is visually observed under a microscope and photographed for evaluation. In addition, the matrigel containing new blood vessels is melted by collagenase treatment in a test tube, the amount of hemoglobin exuded in the solution is measured, and the amount of new blood vessels is measured based on this. In this way, the angiogenesis inhibitory action of the substance added to the Matrigel is evaluated by comparing it with the control (saline-administered group).

[00 3 5] (2) Angiogenesis inhibitory activity CAM (Chorioallantoismembrane) Atlas and rat corneas Atsushi Nintotori Embryo chorioallantoic membrane CAM (Chorioallantois membrane) One of the plates was set up with a disk substrate containing a sample in the rat cornea, and after a few days, The newly formed blood vessels are visually observed under a microscope, photographed, and compared with the control (saline group) for evaluation.

[0036] (3) Inhibition of cancer cell growth and metastasis In vino test using a tumor bearing animal Breast cancer cells or melanoma cells (500,000 200 1) were injected into the skin of SCID mice, and samples ( For example, 1 5.00 mg Z k-, g) is administered every day. After administration, the tumor volume in the mouse skin is observed over time and compared with the control (saline-administered group) mice. At the same time, new blood vessels in the tumor were measured by immunostaining using an antibody against CD31 (PECAM), a blood vessel 'endothelial cell-specific molecular marker, and compared with the control (saline-administered group) mice. I do. On the other hand, the effect of the sample on metastasis of cancer cells was that breast cancer cells or melanoma cells (eg, 500,000 cells / 200 1) were injected from the tail vein of SCID mice and the sample (1500 mg Zkg) was injected every day. 3 to 5 weeks later, the lungs are removed, the number of metastatic lesions found in the lungs is counted under a microscope, and compared with the control (saline-administered group) mice.

[00 37]

The angiogenesis inhibitory activity of this inhibitor may have been confirmed by in vitro and in vivo angiogenesis inhibitory activities other than tube formation tests.

[0038] The inhibitor can take various forms such as a solid such as a powder, a solution, and a suspension. Generally, it is distributed and provided in a solid state such as powder. In addition to the sulfated polysaccharide, the present composition may optionally contain additives in order to enhance the stability of the sulfated polysaccharide. In addition to sulfated polysaccharides, it may contain proteins derived from raw materials for extraction.

[0039] The present inhibitor can be used as a reagent for various researches \ A pharmaceutical composition used for the prevention and treatment of diseases requiring angiogenesis inhibition, It can be used as a food composition or a cosmetic composition for external use that is taken by individuals who have or have a disease or who are at risk.

[0 0 40] (Pharmaceutical composition) The pharmaceutical composition of the present invention contains the inhibitor, and is used for the prevention or treatment of a disease requiring angiogenesis inhibition (hereinafter referred to as the pharmaceutical composition). It can be used as a thing. Examples of diseases requiring angiogenesis inhibition in this pharmaceutical composition include gastric cancer, esophageal cancer, tongue cancer, pharyngeal cancer, colon cancer, lung cancer, liver cancer, kidney cancer, breast cancer, bile duct cancer, knee cancer, prostate Solid tumors such as cancer, endometrial cancer, cervical cancer, ovarian cancer, bladder cancer, skin cancer, hemangioma, multiple bone marrow-aneurysm. '' Also, hemangiofibroma, atherosclerosis, arteriovenous malformation, vascular lesions such as vascular adhesion, granulation, hemangioma, hypertrophic scar, premature aging, psoriasis, scleroderma, keloid, wart and other skin diseases, bleeding Osteoarthritis, non-combination fractures, rheumatoid arthritis, osteoarthritis and other osteoarthritis, ovarian hypertrophy syndrome, follicular cysts, multicystic ovarian genital diseases, age-related macular degeneration, diabetes Examples include pathological retinopathy, neovascular 'tube glaucoma, angiogenesis after cornea transplantation, ocular diseases such as trachoma, lung diseases such as exhaustion and chronic bronchitis. The diseases targeted by this pharmaceutical composition are not limited to these. In addition, the above-mentioned prevention or treatment of various solid tumors includes suppression of metastasis of these various solid tumors.

[0 0 4 1] The sulfated polysaccharide in the pharmaceutical composition may be a free acid in a sulfate ester group or a pharmaceutically acceptable salt. Examples of such salts include alkali metal salts such as sodium and potassium, and alkaline earth metals such as calcium and magnesium. The sulfated polysaccharide of the present invention is preferably a sodium salt.

[0 0 4 2] In addition, the present pharmaceutical composition may contain other drugs effective for the above-mentioned diseases other than the present composition. For example, anti-infective agents such as anti-cancer agents, anti-inflammatory agents, antibacterial agents or antiviral agents can be mentioned. The present pharmaceutical composition may also be provided as a pharmaceutical composition set that can be administered in combination with other drugs effective for the above diseases.

[0 0 4 3] The pharmaceutical composition contains at least the inhibitor as an active ingredient. In addition, various pharmaceutical forms can be prepared including known pharmaceutically acceptable drug carriers. Such a drug carrier is well known in the art, and a known drug carrier used for a pharmaceutical form that may be applied to the present drug composition may be appropriately selected and used. Examples of the pharmaceutical composition of this pharmaceutical composition are 5 powders, powders, granules, tablets, capsules, solids such as chewables, solutions, suspensions, liquids such as emulsions, injections, suppositories, Examples include films and gels. Examples of drug carriers that can be used in such pharmaceutical forms include

-Glucose, lactose, sucrose, starch, mannitol, dex small phosphorus,-glyceride of fatty acid, polyethylene dalcol, hydroxyethylene starch,

10 Ethylene glycol, polyoxyethylene sorbitan fatty acid ester, mino acid, gelatin, albumin, water, physiological saline and the like. Furthermore, additives such as stabilizers, wetting agents, emulsifiers, binders, tonicity agents and the like may be included as necessary. Such a pharmaceutical composition as described above can be prepared by publicly known conventional means corresponding to various preparation forms. ,

15 [0 0 4 4] This pharmaceutical composition contains a mammalian odor: C resistant polysaccharide.

Therefore, even if it is administered orally, it can cause an angiogenesis inhibitory action in the digestive tract of the oral cavity, esophagus, stomach, small intestine, large intestine and the like. In this case, it is provided as various solid or liquid oral preparations. In addition, the pharmaceutical composition can be used as a topical application agent for tumor sites and inflammatory sites. For example, removal of a tumor site

20 Before or after covering the region or inhibiting angiogenesis of a predetermined target site, the present pharmaceutical composition can be used. In addition, this drug composition can be applied to inflammatory sites of rheumatism and various diseases. For example, by injecting and applying this pharmaceutical composition to an inflamed site in the joint, skin, oral cavity, various intima, vascular endothelium, retina, etc., angiogenesis at the inflamed site is inhibited.

25 Can be harmful. The pharmaceutical composition includes, in particular, a liquid preparation, a solid preparation that dissolves at the time of use, a film preparation, a gel preparation, a suppository, an injection and the like as such a preparation form for topical application. In addition, the pharmaceutical composition can be delivered transluminally or transdermally using an endoscope, a catheter, a tube, or the like. Furthermore, this pharmaceutical composition is applied to various indwelling materials such as balloon stents. It may be applied in the form of a tool.

[004 5] The effective dosage of the pharmaceutical composition is determined by the dosage form, administration method, age, weight, symptoms, administration schedule, etc. of the subject, for example, for oral use, sulfated polysaccharide. The dosage is preferably 1 mg to 5000 mg or less per day for adults, more preferably 1 O Omg to 2 000 mg. These may be administered in several divided doses per day. '' In addition, the dose other than oral intake varies depending on the formulation form, administration method, purpose of use and the age, weight, and symptoms of the patient to whom the drug is administered. In the dose of the active ingredient, humans (for example, adults) 0.001 mg body weight to 1000 mg body weight, preferably 0.0 '1 mg Z body weight to 100 mg body weight, more preferably 0. O l mg Z body weight to 30 mg Z body weight. In the case of topical application, the content of sulfated polysaccharide in the pharmaceutical composition is, for example, 100% by weight of the pharmaceutical composition, usually 0.001% to: 100% by weight, preferably Is 0.1 weight. /. ~ 90 wt%, more preferably 1.0 wt% to 800 wt%.

[004 6] (Treatment method)

The therapeutic method of the present invention comprises the step of administering the pharmaceutical composition to an individual having a disease requiring angiogenesis inhibition or an individual having a risk against the disease. In addition, as a dosage form, it can be administered orally to these individuals, or it can be administered locally to such a disease, donor, site or a site at risk for the occurrence of such a disease.

[004 7] (Food composition)

The food composition of the present invention contains the composition and can be used as a food composition (hereinafter referred to as the present food composition) for the prevention, amelioration, or prognosis of diseases that require angiogenesis inhibition. . In the present food composition, the diseases requiring angiogenesis inhibition are the same as those in the present pharmaceutical composition. Further, it may be a free acid at a sulfate ester group in the sulfated polysaccharide contained in the food composition, for example, an alkali metal salt such as sodium or potassium, or an alkaline earth such as calcium. It can be a salt of metal or the like. [0 0 4 8] The food composition may be a general food such as a processed food, a confectionery, a seasoning, a palatability food or a beverage containing the sulfated polysaccharide of the present invention. . Specific forms are not particularly limited, but solid or semi-solid foods such as cookies, biscuits, candy, gum and jelly, fruit juices, teas, coffees, soft drinks and other beverages, breads and potatoes Main foods such as soup, curry, stew, various foods such as sauces, various flavors and seasonings. In addition, dietary supplements, functional foods, special health foods, tube feedings, etc. can be used. As a dietary supplement or the like, it is possible to adopt a preparation form similar to the oral administration form of the above-mentioned pharmaceutical composition.

[0 0 4 9] Further, since the food composition contains the angiogenesis inhibitor of the present invention, the solid tumor, blood vessel lesion, skin disease, bone based on the angiogenesis inhibitory action of the present invention and the action thereof · It may be configured to be used for prevention or treatment of various diseases including arthropathy, genital diseases, eye diseases and lung diseases, as well as metastasis of solid tumors. ,

[0 0 5 0] The effective intake of the food composition depends on the age, weight, and conditions of the subject, but the intake of sulfated polysaccharides is roughly The daily dose is preferably 1 mg to 500 mg or less, more preferably 100 mg to 200 mg. These may be taken in several divided doses per day. '

[0 0 5 1] The content of sulfated polysaccharide in the food composition is not particularly limited, but is preferably about 0.01 to 20% by mass in terms of dry weight, for example. Preferably, it is 0.1 mass% or more and 5 mass% or less.

[0 0 5 2] (Cosmetic composition)

The cosmetic composition of the present invention containing the present composition (hereinafter referred to as the present cosmetic composition) can be used as a cosmetic composition for external use on the skin. The cosmetic composition can be a cosmetic, a quasi-drug, or the like. Specifically, it can be a skin care product such as an emulsion, cream or lotion, or an ointment. [0 0 5 3] The content of the cosmetic composition for external use on the skin is appropriately selected according to the difference in symptoms, for example, 0.01 mass of the total weight. / ₀ or more 5 0 Mass. / About ₀ or less, preferably 0.0 1 mass. / ₀ or more 10 mass. / ₀ or less. This can be applied one to several times in several days. This can be done one to several times a day.

[0 0 5 4] (Pharmaceutical compositions for non-human animals, etc.)

Furthermore, the composition containing the present inhibitor can be used as a pharmaceutical composition or a feed composition for non-human animals. -Non-human animals include livestock, poultry and pets. Even in non-human animals, administration or ingestion of this inhibitor can prevent or treat diseases that require angiogenesis inhibition, and can maintain health. In addition, it is easy for those skilled in the art to produce various forms of pharmaceutical compositions and feed compositions for non-human animals containing this inhibitor.

[Example]

[0 0 5 5] Specific examples will be given below by way of examples, but the present invention is not limited to these examples. "

[0 0 5 6] (Example 1)

(Extraction of sulfated polysaccharides from Hydetasa)

(Sample 1) After swelling 70 5 g (water content of about 7%) of cultured cultivated hitodasa in Mie Prefecture for 30 minutes, the sample was drained for 10 minutes. To this alga body, water 1 8 1 was added to boil, 5.4 g of citrate was added, and hot water extraction was performed at 95 ° C. to 100 ° C. for 6 hours. Water was added to this hot water extract to make a total amount of 18.8 kg, 540 g of diatomaceous earth was added, mixed and centrifuged to obtain a supernatant (15.9 kg). The supernatant liquid 1.51 was further added with 5 g of diatomaceous earth and subjected to vacuum filtration. This operation was repeated, and the total amount of the supernatant was depressurized and filtered, and water was added to the total amount of the filtrate to make 16 kg.

[0 0 5 7] The filtrate was then filtered through an ultrafiltration membrane (molecular weight cut off 10,000). The ultrafiltration treatment is terminated when the membrane permeate is 50%. The concentrated solution was supplemented with a reduced amount (permeate amount) to a total volume of 16 kg. This place The process was repeated a total of 3 times. The finally obtained 16 kg of concentrated liquid was spray-dried to obtain a powder (sample 1: 2700 g).

[0 0 5 8] GPC was performed on Sample 1 under the following HP L C conditions. According to GPC, the weight average molecular weight in terms of pullulan of Sample 6 was 510,000. 5 HPLC conditions:.

Column: SHODE x OH Pak S B— 8 0.6M HQ 2 Eluent: 0.1 M N a C 1

-^ Power ram temperature: 4 0 ° C — — —

'Detector: R I'

10 Flow rate: 1 m 1 / min

[0 0 5 9] (Sample 2). After swelling 750 g (water content: approx. 7%) of Mie Prefecture-cultivated sardine in 18 1 water for 30 minutes, drain for 10 minutes. The This operation was performed a total of 3 times to wash the algal cells. Water was extracted from this algal body with hot water at 95 ° C to 100 ° C for 6 hours. Add water to this hydrothermal lysate to a total volume of 18.8 kg, add 10 8 pg of diatomaceous earth, mix and centrifuge to obtain a supernatant (13.6 kg). It was. The whole amount of the supernatant was spray-dried to obtain a powder (sample 2 ′: 140 g). When GPC was performed under the same conditions as in Sample 1, the pullulan equivalent weight average molecular weight of Sample 2 was 250,000. '

20 [0 0 60] (Samples 3 to 5) About 6 g of Sample 1 was dissolved in 3M 0. 05MKG 1 solution 30.000 ml (2% solution). This solution was injected into an ion-exchange column DE 52 packed column (manufactured by Nikkoman, diameter 50 mm, length 500 mm), and 7 M urea 0.05 MK as the mobile phase. 500 ml of C 1 solution, 0.1 M of 7 M urea 0.15 MK of C 1 solution, 300 ml of 7 M urea, 0.225 2 5 MKC 1 solution of 1 500 ml and 7 M urea 2 The MK C 1 solution was passed through 1500 ml, and the polysaccharide was fractionated by gradually increasing the KC 1 concentration. The elution fraction at the time of passing 7 M urea 0.25 MKC 1 solution was dialyzed for 4 to 5 days under running water using a cellophane membrane, and the concentrated solution was lyophilized to give Sample 3. In addition, dialyze the elution fraction at the time of 7 M urea 2 MK C 1 passage and freeze the concentrate. It was set to sample 4 after drying. Furthermore, 1 g of elution fraction at the time of 7 ^ urea 0.25 MKC 1 solution flow was dissolved in 50 m 1 of distilled water, and using a cation exchange resin SK 104 packed column (Mitsubishi Kasei Co., Ltd.). The weak thione solution was recovered (pH 1-9 to 2.4), neutralized with INNaOH, and freeze-dried to obtain a powder (sample 5). For Samples 3 and 4, GPC was performed under the same conditions as Sample 1. The pullulan equivalent weight average molecular weights of Samples 3, 4, and 5 were 11.9 million, 230,000, and 220,000, respectively.

[0 0 6 1] (Sample 6) Mie Prefecture-cultivated chickpeas 24 liters in 6 1 water, soak for 30 minutes, separate the wash water and add water again to make a total volume of 6. 24 kg It was. This was subjected to hot water extraction for 2 hours while maintaining the liquid temperature at 60 ° C to 65 ° C. Centrifugation gave a hot water extract of 3.79 9 g and solids (hereinafter referred to as “recovered water”) 1. 2 3 kg. To 300 g of this recovered hitodasa, water 31 and 100 mg of guenoic acid were added, and hot water extraction was performed while adding water at 95 ° C to 100 ° C for 6 hours in a timely manner. The hot water extract was filtered, and the entire amount of the filtrate was freeze-dried to obtain a powder (3 g). Dissolve the powder in 0. 0 ml of 0.1 mM Tris buffer (pH 8.7) and add about 0.14 g of proteolytic enzyme (actinase-1 from Kaken Pharmaceutical Co., Ltd.) Incubate for 15 hours at 50 ° C, inject into a DE 5 2 packed column (equilibrated with 71 ^ urea 0.1 5M KC 1), pass 7M urea 0.1 5MKC 1 3 1 or more, The KC 1 concentration was passed through the gradient from 0, 15 M to 2 M over about 11 hours, and each fraction eluted was collected with a fraction collector. GPC was performed on the various fractions under the same conditions as GPC performed on Sample 1, and the fraction with a retention time (RT) of about 14.5 minutes as the main peak was used as the recovered fraction. In summary, this whole amount was dialyzed for 4 to 5 days using a dialysis tube, and the concentrate was freeze-dried to obtain a powder (sample 6). For Sample 6 thus obtained, GPC was performed under the same conditions as Sample 1. Sample 6 had a pullulan equivalent weight average molecular weight of 1,890,000.

[0 0 6 2] (Sample 7) 3550 g of powder was obtained by freeze-drying 5500 g of the hot water extract of the hot water extract obtained at the time of producing Sample 6. Add 3 g of this powder to 0.1 mM Dissolve in squirrel buffer (pH 8.7) in 20 ml, add 0.14 g of proteolytic enzyme (Kachinase E, manufactured by Kaken Pharmaceutical Co., Ltd.) and incubate for 15 hours at 50 ° C. After that, inject into DE 5 2 filling force ram (equilibrated with 7 M urea 0.15 MKC 1), and after passing 7 M urea 0.1 5 MKC 1 3 1 or more, K C.1 concentration 0.1 Gradient was applied for about 11 hours from 5M to 2M, and each elution fraction was collected with a fraction collector. For the various fractions, call out the peak with a retention time (RT) of about 15.5 minutes under the GPC conditions for sample 1 and call the fraction with the main peak as the recovery flag. Then, dialysis was performed for 5 days using a dialysis tube, and the concentrate was freeze-dried to obtain a powder (sample 7). Sample 7 thus obtained was subjected to GPC under the same conditions as Sample 1. The weight average molecular weight in pullulan conversion of Sample 7 was 620,000.

[0 0 6 3] (Sample 8) Dissolve 3 g of Sample 1 in 200 ml of O. l mM Tris buffer (pH 8.7). Proteolytic enzyme (actinase E manufactured by Kaken Pharmaceutical Co., Ltd.) 0. 1.4 g added, 50 ° C; C 15 incubated for 5 hours, then injected into DE 5 2 filling force ram (equilibrated with 7 M urea 0.15 MKC 1), 7 M urea 0.1 5MKC 1 3 After passing 1 or more, 1 :. Gradient flow was carried out at a concentration of 0.1 to 5 M to 2 M over about 11 hours, and each fraction eluted was collected with a fraction collector. For the various fractions, the fraction with a retention time (RT) of approximately 17 minutes under the GPC conditions for sample 1 as the main peak is collected as the collected fraction, and this total amount is collected using a dialysis tube. Dialyzed for a day and freeze-dried the concentrate to obtain a powder (sample 8). For sample 8 thus obtained, GPC was performed under the same conditions as for sample 1. Sample 8 had a pullulan equivalent weight average molecular weight of 210,000.

[0 06 4] (Sample 9) 3 g of powder was obtained by freeze-drying 50 g of the hot water extract of hot water from the sample 6 produced at the time of sample 6 production. 3 g of this powder was used as sample 9. The weight average molecular weight in terms of pullulan according to GPC of sample 9 was 1 million.

[00 6 5] (Sample 1 0) 400 g (water content of about 7%) The sample was immersed in 10 1 water for 30 minutes to swell, and then drained for 10 minutes. To this algae, add water 1 0 1 and boil, add 28.8 g malic acid,

9 Hot water extraction was performed at 5 ° C to 100 ° C for 4 hours. Water was added to this hot water extract to adjust the total amount to 10 kg, and 2.5 kg was subjected to solid-liquid separation by diatomaceous earth filtration. This clear solution (30 O g) was desalted and purified with a dialysis tube, and then lyophilized. 3 g of lyophilized product is dissolved in 200 ml of 0.1 mM Tris buffer (pH 8.7), and 0.14 g of proteolytic enzyme (actinase E manufactured by Kaken Pharmaceutical Co., Ltd.) is added. After 1-hour incubation at 0 ° C, inject into DE 5 2 filling force ram, (equilibrated with 7M urea 0.1 5M KC 1) and 7M urea 0.1 5 MKC 1 3 1 After the above flow, the 1: 0.1 concentration was passed through the gradient from 0.1. 5 M to 2 M over about 11 hours, and each elution fraction was collected with a fraction collector. For the various fractions, the fraction that has a retention time (RT) of about 18 minutes under the GPC conditions for sample 1 as the main peak is collected as the recovered fraction, and this total amount is collected using a dialysis tube. The mixture was dialyzed for 5 days, and the concentrated solution was freeze-dried to obtain 2 g of powder (sample 10). When GPC was performed on Sample 10 under the same conditions as Sample 1, the weight average molecular weight in terms of pullulan was 50,000.

[00 6 6] (Example 2)

(Evaluation of angiogenesis inhibitory activity)

Samples 1 to 8 prepared in Example 1 were evaluated for angiogenesis inhibitory activity using human umbilical cord-derived vein endothelial cells (HUVEC) as an index of lumen formation. First, after rapidly adding 0.3 ml of 10 mg / m 1 concentration of gel rig (manufactured by BD 1) in ice-cooled 24well plates, 5% C 02 Gelation was performed by incubating in an incubator (37 ° C) for 2 hours.

[006 7] Next, HUVE C was seeded in a collagen-coated petri dish,

Using MCDB 1 3 1 culture solution (manufactured by Chlorella Kogyo Co., Ltd.) containing 10% tussive fetal serum (FBS), logarithmic growth is performed from middle to late in a 5% C2 incubator at 37 ° C. Until cultured. Then with phosphate buffered saline After washing twice, it was treated with 0.05% trypsin ^/ 0.02% EDTA for about 1 minute to separate and float HUVEC from the petri dish wall. An appropriate amount of 10% FBS-containing MCDB 13 1 culture solution was added and pipetted to prepare a HUVEC floating solution. After counting the number of HUVE C cells with a hemocytometer, it was diluted with 5% FBS-containing MC DB 1 3 1 culture solution to finally prepare a cell suspension of 2 × 10 4 cells Zm 1. Then the sample in the HU VE C suspension to a final concentration of 0, 0.0 1, 0.1 and 1 xg / ml :! -8 were added, and this -HU VEC suspension was inoculated onto a gelled Matrigel and cultured at 37 for 6 hours in a CO ₂ incubator. Then, observe with a phase-contrast microscope (Olympus, magnification: 1.000 times), take an image with a CCD power camera, and use image analysis equipment (N IH I.MA GE) for at least 4 fields of view. The length of the lumen contained in each field of view was measured, and the average value was taken as the length of each lumen. From the lumen length thus obtained and the lumen length similarly measured in the absence of sulfated polysaccharide, the lumen formation inhibition rate (%) ′ was calculated from the following equation. The results are shown in Figs.

[0 0 6 8] Tube formation inhibition rate (%) = (Lumen length in the absence of sulfated polysaccharide) Length of sample in one sample) (Lumen length in the absence of sulfated polysaccharide) X 1 00

[0 6 6] As shown in Fig. 1 and Fig. 2, all samples inhibited the formation of lumens, especially in Sample 2, Sample 5, and Sample 6, compared to the control. The tube formation was inhibited to less than%. From these facts, any of the sulfated polysaccharides in Samples 1 to 8 has angiogenesis inhibitory activity. Among them, the sulfated polysaccharide fractions in Sample 2, Sample 5 and Sample 6 have high angiogenesis inhibitory activity. It turns out that there is sex. In particular, sample 2, which is a crudely purified fraction and has a maximum weight average molecular weight (2500,000) in terms of pullulan in GPC, showed high angiogenesis inhibitory activity. In Samples 6, 7 and 8, it was found that the higher the average molecular weight in GPC, the higher the inhibitory activity. On the other hand, in the low molecular weight samples 4 and 5 having a weight average molecular weight of 220,000 to 230,000 in GPC, It was found that Sample 5, which was desalted by ion exchange and then adjusted to sodium single salt, showed higher angiogenesis inhibitory activity than Sample 4 before ion exchange treatment.

[0 0 7 0] (Example 3)

For samples 1, 2, 5-8 and 1: 0, the sulfuric acid content and the component ratio of the constituent monosaccharides were measured. In addition, each was measured by the following method. The results are shown in Table 1.

[0 0 7 -1] (1) Method for measuring sulfate group content.

, (Combustion flask method)

Elemental Analysis of Sulfur Atoms, Analytical Chemistry, Volume 1, 6 8 9-6 9 1 (1 9 6 6), Volume 7, 1 3.2: 1 3 24: (1 9 6 8 The sulfate group content was calculated by multiplying the sulfur atom content (weight) in the obtained sample by 3. '

[00 7 2] (2) Measuring method of constituent monosaccharides

Acid hydrolysis of the sample (dissolve the sample (approximately 20 mg) in 2.N sulfuric acid (1 O ml) and heat in a boiling water bath for 2 hours), and then perform high performance liquid chromatography (column: A sahipack NH ₂ P- Various monosaccharides are identified by 50 4 E, power ram temperature: 35 ° C, mobile phase: water Z acetonitrile, flow rate: lm 1 / min, detector: RI) And quantified.

[00 7 3]

[table 1]

ND: not detected

[0 0 74] As shown in Table 1, Samples 1, 2, 5-8 and 10 all have rhamnose as the main component of the constituent monosaccharide, and the mass ratio is 6 8 mass%. To 93% by mass, and 86% by mass with the mass ratio of glucose, the second main component. / ₀ to 9 7 mass. / Over to ₀ and further added xylene opening one scan of the mass ratio was 9: 2% to 1 0 0 mass%. In Sample 2, Sample 5 and Sample 6 having good angiogenesis inhibitory activity, the mass ratio of rhamnose was in the range of 75 mass% to 93 mass%. As for these 3 'species of the sample, the mass ratio of glucose, 1 1 quality from 4 wt% - a冓^_0/0, further, the mass ratio of xylose, a 7 wt% to 3 wt% It was. Incidentally,. The sum of these three samples, the mass% of the mass ^_0/0 and Dal course rhamnose, 8 6 wt. /. It was found that these two types of monosaccharides constitute the majority of monosaccharides.

[0 0 7 5] Also, as shown in Table 1, in Sample 2, RhamZGul ratio (mass of rhamnose ./. Roh mass ^_0/0 glucose) force 6. a 8, Sample 5, 2 3. 3, Sample 6 'is 18.2. The Rham / Xylo ratio (rhamnose mass% xylose mass ⁰ /.) Is 10.7 for sample 2, 3 1. では for sample 5, and 22.8 for sample 6. (Rham + Gul ) / Xylo ratio (mass percentage of total noxylose of rhamnose mass ⁰ /. And glucose mass ⁰ /.) Is 1 2.3 for sample 2 and 3 2, 3 for sample 5 6 was 24. .

[00 7 6] (Example 4)

In the present example, the effect of the extract component on the metastasis of cancer in vivo was evaluated by the following method.

[00 7 7] Measurement of cancer cell metastasis in vivo

Using 10 C57 / BL6 mice in each group, the effect of the hot water extract (sample 2 of Example 1) on cancer cell metastasis was measured. B16 mouse melanoma cells (500,000 / 200μ1) were injected through the tail vein of mice, and a hot water extract (sample 2 of Example 1) (250 mg / kg) was orally administered every day. After a week, the lungs were removed, and the number of cancer cell metastases found in the lungs was examined under a microscope. By counting and comparing it with that of a control (distilled water administration group) mouse, the effect of a hot water extract (sample 2 of Example 1) on metastasis of cancer cells was evaluated. The results are shown in FIGS. The transfer rate is the% of the number of metastases in the hot water extract from the number of metastases in the control.

[0 0 7 8] As shown in Figs. 3 and 4, B16 mouse melanoma cells injected from the tail vein of C57 / BL6 mice were observed to metastasize to the lung after 2 weeks. The number was significantly and significantly lower in the oral administration of the hot water extract of the hot water extract than in the group administered with distilled water.

[0 0 7 9] From the above, it was found that the hot water extract of Hitoedasa significantly suppressed lung metastasis of cancer cells in vivo.

Claims

The scope of the claims

1. An angiogenesis inhibitor comprising a sulfated polysaccharide or a salt thereof containing rhamnose as a main constituent monosaccharide.

2. The inhibitor according to claim 1, wherein the sulfated polysaccharide has the following characteristics.

-(a) In the total mass of the constituent polysaccharide, the rhamnose content is 50 mass% or more,-the glucose content is 1 mass% or more and 30 mass% or less, and the xylose content is 10 mass% or more and 10 mass% or more. /. Less than

(b) Sulfuric acid group content of 10% to 40% by mass,

3. The angiogenesis inhibitor according to claim 1 or 2, wherein the weight average molecular weight in pullulan conversion by gel filtration chromatography is from 50,000 to 300,000. '

15 4. The sulfated polysaccharide is derived from an algae belonging to the moss algae. The inhibitor according to any one of to 3.

5. The inhibitor according to any one of claims 1 to 4, wherein the inhibitor contains the algae or an extract thereof.

6. The inhibitor according to claim 5, wherein the extract is a hot water extract of the algae. ,

7. The inhibitor according to any one of claims 4 to 6, wherein the algae is selected from algae belonging to the genus Monostroma.

8. The inhibitor 25 according to the range 7 of the wording solicitation, which is the order ij diatom algae f, Mon os t ro m a La t i s s i m u m or ί o M o n s t r oman t i i t i d um.

9. The inhibitor according to any one of claims 1 to 8, wherein the sulfated polysaccharide is a sodium salt.

1 0. An angiogenesis inhibitor comprising an algae belonging to the genus Monostroma or an extract thereof.

1 1. The angiogenesis inhibitor according to claim 10, comprising a sulfated polysaccharide or a salt thereof containing rhamnose as a main component in charge.

1 2. A pharmaceutical composition for preventing or treating a disease requiring angiogenesis inhibition, comprising the angiogenesis inhibitor according to any one of claims 1 to 11. .

1 3. The pharmaceutical composition according to claim 12, wherein the disease is selected from the group consisting of solid tumors, vascular lesions, skin diseases, osteo / arthropathy, genital diseases, eye diseases and lung diseases.

1 4. The medicinal IJ composition according to claim 12, wherein the disease is solid tumor metastasis.

1 5. The pharmaceutical composition according to any one of claims 12 to 14, wherein the pharmaceutical composition is for topical administration. ·

1 6. The pharmaceutical composition according to any one of claims 12 to 14, which is for oral administration. .

1 7. A food composition for preventing or prognosing a disease requiring angiogenesis inhibition, comprising the angiogenesis inhibitor according to any one of claims 1 to 11.

1 8. The food composition according to claim 1'7, wherein the food composition is a nutritional supplement composition. '

1 9. The food composition according to claim 17 or 18, wherein the food composition is used for inhibiting metastasis of solid tumor.