CN101001617A - Antitumor agent - Google Patents
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- CN101001617A CN101001617A CN 200580025889 CN200580025889A CN101001617A CN 101001617 A CN101001617 A CN 101001617A CN 200580025889 CN200580025889 CN 200580025889 CN 200580025889 A CN200580025889 A CN 200580025889A CN 101001617 A CN101001617 A CN 101001617A
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Abstract
An antitumor agent, an apoptosis-inducer or an angiogenesis inhibitor which contains a compound contained in a less expensive edible basidiomycetes or a composition originating in the edible basidiomycetes and can exhibit its effects even by oral administration; and a food containing the above-described compound or composition which has an antitumor activity, an apoptosis-inducing activity or an angiogenesis inhibitory activity. These products are useful particularly in the fields of drugs and health foods.
Description
Technical field
The present invention relates to from the mushroom physiologically active ingredient as antitumor agent, programmed cell death inducing agent or the angiogenesis inhibitor of effective ingredient and the food that contains this composition.
Background technology
The edible basidiomycetes that makes preparations for sowing such as extensive known Lentinus Edodes (Lentinus edodes), Grifola frondosa (Grifola frondosa), JINZHENGU (Flammulina velutipes) has antitumor activity, and their sporophore or mycelium can prepare various antitumor activity compositionss up to now.Clearly edible basidiomycetes a kind of also---elm is living (lyophyllumulmarium) to have an antitumor activity from pleat umbrella (Block Na シ メ イ ヅ), the sporophore hot water extract and the purified polysaccharide body (for example patent documentation 1) thereof that for example clearly will have 3-20% protein content, 20-50% sugar content are suspended in water or the hydrophilic solvent, pass through heat treated then, obtaining molecular weight is 6,000-60,000 biological active substances EEM-S, it has anti-tumor activity (for example patent documentation 2).
Known malignant tumor forms and propagation can generate by induction of vascular.Angiogenesis is induced, and then can transport nutrition to tumor, makes tumor cell propagation and deterioration rapidly, and perhaps tumor cell is invaded in the blood vessel, moves in the various tissues, shifts easily.That is,, then can suppress propagation, deterioration or the transfer of tumor cell if suppress angiogenesis.Developed a lot of medicines based on above-mentioned conception, but the commercialization of still being unrealized at present.
On the other hand, report about the material of the programmed cell death of induced tumor cell is arranged a lot.But the material that brings out programmed cell death can be target with the endotheliocyte of the high new vessels of multiplication capacity also.Someone has reported the material (for example non-patent literature 2) based on the inhibition tumor of above-mentioned viewpoint exploitation.Above-mentioned substance causes a little less than the effect of programmed cell death for tumor cell itself, but can suppress the inductive angiogenesis of tumor cell, promptly has the effect of attacking its army provisions, so it is little to infer its side effect.
Have report to point out: the composition from mushroom has the effect that suppresses angiogenesis.For example from the ergosterol (non-patent literature 3) of Brazilian mushroom (Agaricus blazei Murill).
Terpene compound (terpenoid) is the organic compound that is present in a lot of plants, and carbon number is 5 multiple, the precursor substance that comes a free n isoprene or isopentane to constitute.N=2 is a terpenes, and n=3 is a sesquiterpene, and n=4 is two terpenes, and n=5 is a sesterterpene, and n=6 is a triterpene, and n=8 is a tetraterpene, and this is above is called polyterpene.Known terpene compound has the effect of bringing out programmed cell death, and a lot of reports of bringing out the programmed cell death effect about triterpene etc. are for example arranged.
Known living in the physiologically active ingredient of pleat umbrella from elm, the SBS material has effect and the anticancer promoter effect (patent documentation 3) that suppresses platelet aggregation.Also known elm is living to contain various polyterpenes (for example non-patent literature 1) as bitter principle from the pleat umbrella.
Patent documentation 1: the spy opens flat 5-306233 number
Patent documentation 2:WO 01/51070
Patent documentation 3: the spy opens flat 4-104795 number
Four-players such as non-patent literature 1:Sawabe A., Journal of Mass Spectrometry,, Vol.31,921-925 page or leaf in 1996
Two people such as non-patent literature 2:Schwarz R.E., J.Surg.Res.,, Vol.120, No.1,64-72 page or leaf in 2004
Three people such as non-patent literature 3:Takaku T., J.Nutr., calendar year 2001, Vol.131, No.5,1409-1413 page or leaf
Summary of the invention
The sporophore of clear and definite various edible basidiomycetes has antitumor activity, but in order to obtain effect, must daily edible certain amount.But edible basidiomycetes also has special character aspect taste, and is even handled in culinary art, daily edible a certain amount of also very difficult.
On the other hand, circulating extensively the health food that has cooperated the effective ingredient with antitumor activity, immunocompetence etc. that is obtained by certain basidiomycetes on the market, still as the basidiomycetes rareness of raw material, the health food of production is also just very expensive.
Even the object of the present invention is to provide cheap, as to be used to the oral also display effect made from the chemical compound of the tool anti-tumor activity of edible basidiomycetes medicine or food.
The inventor has carried out research in depth for solving above-mentioned problem, found that: the polyterpene from the living sporophore from the pleat umbrella of elm has anti-tumor activity, programmed cell death brings out activity and angiogenesis inhibiting activity, thereby has finished the present invention.
Promptly, put it briefly, first invention then of the present invention relates to antitumor agent, programmed cell death inducing agent or angiogenesis inhibitor, it is characterized in that: contain and be selected from least a as effective ingredient of chemical compound, its derivant and the acceptable salt of pharmacology shown in the following general formula (1).
(in the formula, m represents the integer of 1-9, and n represents the integer of 1-7, m+n=5～11, R represents hydrogen atom or hydroxyl).
Second invention of the present invention relates to antitumor agent, programmed cell death inducing agent or the angiogenesis inhibitor of first invention, and wherein, the chemical compound shown in the general formula (1) is at least a chemical compound shown in following formula (2)-(11).
The 3rd invention of the present invention relates to food, this food has anti-tumor activity, programmed cell death brings out activity or angiogenesis inhibiting activity, it is characterized in that: contain and be selected from the chemical compound shown in the general formula (1), its derivant and their pharmacology and go up at least a of acceptable salt.
The 4th invention of the present invention relates to the food of the 3rd invention, and wherein, the chemical compound shown in the general formula (1) is at least a chemical compound shown in formula (2)-(11).
The 5th invention of the present invention relates to antitumor agent, programmed cell death inducing agent or angiogenesis inhibitor, it is characterized in that: contain from the living compositions from the pleat umbrella of elm as effective ingredient, described compositions contains chemical compound, its derivant and their pharmacology shown in the formula of being selected from (2)-(11) and goes up at least a of acceptable salt.
The 6th invention of the present invention relates to antitumor agent, programmed cell death inducing agent or the angiogenesis inhibitor of the 5th invention, it is characterized in that: from the living compositions from the pleat umbrella of elm is that the living ethyl acetate extract from the pleat umbrella of elm is adsorbed in silicagel column, then the compositions that obtains of eluting.
The 7th invention of the present invention relates to food, this food has anti-tumor activity, programmed cell death brings out activity or angiogenesis inhibiting activity, it is characterized in that: contain from the living compositions from the pleat umbrella of elm, described compositions contains chemical compound, its derivant and their pharmacology shown in the formula of being selected from (2)-(11) and goes up at least a of acceptable salt.
The 8th invention of the present invention relates to the food of the 7th invention, and it is characterized in that: from the living compositions from the pleat umbrella of elm is that the living ethyl acetate extract from the pleat umbrella of elm is adsorbed in silicagel column, then the compositions that obtains of eluting.
The 9th invention of the present invention relates to from the living compositions from the pleat umbrella of elm, said composition contains chemical compound, its derivant and their pharmacology shown in the formula of being selected from (2)-(11) and goes up at least a of acceptable salt, be that the living ethyl acetate extract from the pleat umbrella of elm is adsorbed in silicagel column, eluting obtains then.
The tenth invention of the present invention relates to suffering from needs the treatment of diseases of antitumor action, programmed cell death fall out effect or angiogenesis suppression action method, and this method comprises: at least a body that tried of suffering from this disease of chemical compound, its derivant and their pharmacology shown in the above-mentioned general formula of being selected from of effective dose (1) being gone up acceptable salt.
The 11 invention of the present invention relates at least a application in antitumor agent, programmed cell death inducing agent or angiogenesis inhibitor preparation that is selected from the last acceptable salt of the chemical compound shown in the above-mentioned general formula (1), its derivant and their pharmacology.
The effect of invention
According to the present invention, even can obtain cheap, contain chemical compound contained in the edible basidiomycetes or from oral antitumor agent, programmed cell death inducing agent or the angiogenesis inhibitor that also has effect of compositions of edible basidiomycetes, and contain this chemical compound or compositions, have anti-tumor activity, food that programmed cell death brings out activity or angiogenesis inhibiting activity.
The accompanying drawing summary
Fig. 1 represents the HL-60 cell in not adding the IPMI-1640 culture medium of FCS, at 37 ℃ of cultivations 6 hours, the figure of the programmed cell death fall out effect of acetone elution fraction.Among the figure, carrier is represented 1% ethanol water.
The best mode that carries out an invention
Occurring in nature, elm is living to grow thickly on the withered tree various broad leaf trees in the fall the time or from life, compares with other mushroom from the pleat umbrella, has the good meat of shapeliness or mouthfeel, can be used as delicious mushroom and gathers edible.In recent years, set up and used the culture medium that in sawdust, cooperates Testa oryzae or other nutrient source,, stably gathered in the crops mushroom with can haveing nothing to do season the whole year with the bacterium bed artificial culture method that bottle or case are cultivated.That is, raw materials of effective components elm of the present invention is living can be obtained cheaply from the pleat umbrella, is suitable as medicine or health food raw material.
Living as the elm of raw material use can be that natural prodcuts also can be the artificial culture products from the pleat umbrella, preferred elm living from pleat umbrella (Lyphyllum ulmarium) M-8171 (FERM BP-1415, preservation day: on August 23rd, 1986), or elm living from pleat umbrella (Lyphyllum ulmarium) K-0259 (FERM P-12981, preservation day: on June 2nd, 1992) (these bacterial strains all be deposited in Independent Administrative Leged Industrial Technology Complex Inst specially permit biological sustenance center (
305-8566 builds the Hitachinaka County, Japan a kind of ground of 1 fourth order, east, ripple city l central authorities the 6th)).These bacterial strains are also sold on market with trade name " ゃ ま び こ ほ ら め じ (registered trade mark) " or " Super ゃ ま び こ ほ ら め じ (registered trade mark) ".Sporophore can be bright product, also can be by the dry thing of exsiccant sporophore such as heat drying, sun dehydration, lyophilization.Sporophore can be to use as raw material with whole strain or with the form of pulverizing.Raw material can also use elm living mycelium or its lyophilization thing from the pleat umbrella.
The inventor found that for studying from the living material with anti-tumor activity from the pleat umbrella of elm: general formula (1) even shown in polyterpene have oral intensive anti-tumor activity, the programmed cell death that also can take effect and bring out activity and angiogenesis inhibiting activity.
The example of chemical compound has chemical compound shown in formula (2)-(11) shown in the general formula that uses among the present invention (1), from the living content angles considerations many, preparation easily from the pleat destroying angel of elm, especially preferably uses chemical compound shown in the formula (3).
As the novel substance SBS of chemical compound shown in the formula (3) of an effective ingredient part of the present invention and Japanese kokai publication hei 4-104795 number is same compound (following chemical compound shown in the formula (3) is called SBS), put down in writing in this patent, SBS has the activatory effect of the Epstein-Barr virus of inhibition, so this chemical compound has anticancer promoter effect.But, of the present inventionly orally can not show that anti-tumor activity, programmed cell death bring out activity and angiogenesis inhibiting activity even show in this communique yet.Promptly, found preventative activity for Japanese kokai publication hei 4-104795 number for the generation of tumor, and the present invention is as described later shown in the embodiment 1, found SBS have treatment for cancer by oral administration or suppress cancer progression, transfer effect, be invention about the novel use of SBS.In addition, undertaken finding in the analytic process by aftermentioned embodiment the inventor, people's such as the chemical constitution of SBS and damp limit document (four-player such as sawabe A., Journal of MassSpectrometry, 1996, Vol.31,921-925 page or leaf) the hypsiziprenol A9 of record is a same substance.
Preparation method about chemical compound shown in the formula of using among the present invention (1) is not particularly limited, can prepare by chemosynthesis or by known method such as extractions in the natural product (for example above-mentioned elm is living from the pleat umbrella), when for example preparing SBS, can adopt Japanese kokai publication hei 4-104795 number method; During chemical compound shown in preparation formula (2)-(11), can be according to the method preparation of people's such as above-mentioned damp limit document.Can also be by the method for following preparation example 1, based on extract obtained, by chemical compound shown in various chromatograph productions (2)-(11).
For example hydrolysis easily in vivo such as esters of the derivant of chemical compound shown in the general formula that uses among the present invention (1), can bring into play the derivant (prodrug) of wishing required effect.Giving the derivant that mammal and metabolism obtain with chemical compound of the present invention is also contained in the derivant of the present invention.The preparation of described prodrug can be carried out according to known method.Described derivant can be their salt.
In addition, the salt of chemical compound or derivatives thereof shown in the general formula (1) can use the pharmacology to go up acceptable salt.Prodrug can be the derivant that can bring into play this chemical compound of function.In addition, various isomers such as the optical isomer of the chemical compound that the present invention uses, ketone-pure tautomer, geometric isomer, the separator of each isomer as long as have anti-tumor activity, programmed cell death brings out activity or angiogenesis inhibiting activity, all can use in the present invention.
The pharmacology of chemical compound shown in the general formula (1) goes up acceptable salt for example alkali metal salt, alkali salt, with the salt of organic base etc.The pharmacology who uses among the present invention goes up acceptable salt and is meant biology nontoxicly in fact, and has the salt that anti-tumor activity, programmed cell death bring out the chemical compound of activity or angiogenesis inhibiting activity.This salt for example has sodium, potassium, calcium, magnesium, ammonium or protonated benzyl star (N, N '-two-benzyl ethylenediamine), choline, ethanolamine, diethanolamine, ethylenediamine, meglumine (N-methylglucosamine), benethamine (N-benzyl-1-phenylethylamine), piperazine or trometamol salt such as (2-amino-2-hydroxymethyl-1, ammediols).The preparation of described salt also can be carried out according to known method.
In the 5-8 of the present invention invention, use contain be selected from chemical compound, its derivant and the acceptable salt of pharmacology shown in above-mentioned formula (changing 2)-(the changing 11) at least a, from the living compositions of elm from the pleat umbrella.Said composition is not particularly limited, and for example can be the living ethyl acetate extract from the pleat umbrella of elm is adsorbed on the silicagel column, then the compositions that obtains by suitable eluting solvent eluting.
For example use elm living when pleat umbrella fresh sporophore, the extracting operation that is used to obtain the living ethyl acetate extract from the pleat umbrella of above-mentioned elm is with respect to 1 weight portion fresh sporophore, preferred 1-10 weight portion, the more preferably ethyl acetate of 1-5 weight portion used, when using sporophore drying thing, with respect to the dry thing of 1 weight portion, preferably use 5-50 weight portion, the more preferably ethyl acetate of 10-25 weight portion.The ethyl acetate boiling point is low, and can be used as the extractant of food, in the secure context excellence.Extraction temperature is not particularly limited, for example be 5 ℃ near the temperature the ethyl acetate boiling point (77.1 ℃), preferred 10-70 ℃, further preferred 15-65 ℃.The extraction time is not particularly limited, and for example 30 minutes-24 hours, preferred 1-20 hour, further preferred 2-10 hour.Extraction is handled to leave standstill and is carried out, and also can under agitation carry out.
By removing insoluble matter in the above-mentioned gained acetic acid ethyl ester extract, this can carry out according to conventional method, for example can be by filtering or the centrifugal insoluble matter of removing.Here, insoluble matter is meant when filtering by the insoluble composition in the extract of commercially available filter paper (for example No.2 filter paper of Advantec manufacturing) filtering, if centrifugalize, be meant the insoluble composition in the extract that the centrifugal force precipitation by at least 5000 * g obtains.
Silicagel column is not particularly limited, and for example can use to be filled with the post of particle diameter as 0.063-0.200mm silica gel.To wherein supplying with above-mentioned acetic acid ethyl ester extract, as required with the ethyl acetate washing, use all kinds of solvents eluting then then.Eluting solvent is not particularly limited, and for example can use acetone.
The gained eluent can directly be used as compositions of the present invention, for example this eluent drying under reduced pressure can also be solidified, and dissolves with ethanol then again, solidifies by drying under reduced pressure and makes compositions of the present invention.The content of chemical compound is not particularly limited shown in said composition Chinese style (2)-(11), preferred 0.001-100 weight %, more preferably 0.01-10 weight %.Chemical compound shown in formula (2)-(11) is adjusted to desired concn, and its method is not particularly limited, and can add excipient such as dextrin.Wherein, the content of chemical compound is 0.0001-10 weight %, more preferably 0.001-1 weight % shown in the preferred said composition Chinese style (3).
With chemical compound shown in the general formula (1) that uses in antitumor agent of the present invention, programmed cell death inducing agent or the angiogenesis inhibitor, its derivant, the acceptable salt of pharmacology or above-mentionedly be called effective ingredient of the present invention from the living compositions from pleat umbrella compositions of elm, antitumor agent, programmed cell death inducing agent or the angiogenesis inhibitor that will contain effective ingredient of the present invention are called medicine of the present invention.
As described later, effective ingredient of the present invention is not seen toxicity.In addition, also need not to worry the generation of side effect.Therefore, can be safely and express anti-tumor activity aptly, programmed cell death brings out activity and angiogenesis inhibiting activity.
Among the present invention, as the object disease of antitumor agent so long as need the disease of antitumor action to get final product in the treatment, be not particularly limited, for example all malignant tumor, for example gastric cancer, colorectal cancer, esophageal carcinoma, skin carcinoma, uterus carcinoma, carcinoma of prostate, bladder cancer, pulmonary carcinoma etc. can be used as above-mentioned disease.And, also can be used as the object disease of antitumor agent of the present invention as hemopoietic organ's such as leukemia malignant tumor.Anti-tumor activity can followingly be measured: add medicine in cultivating cancerous cell, measure the survival rate of cancerous cell after the cultivation stipulated time, perhaps with cancer cell transplantation in laboratory animals such as mice, with medicine give certain during, the size of the entity tumor that produces in the determination experiment animal body or weight etc. then.
Among the present invention, as the object disease of programmed cell death inducing agent,, be not particularly limited so long as in treatment, need the disease of programmed cell death fall out effect to get final product, above-mentioned all malignant tumor are for example arranged, in addition also have autoimmune disease such as sjogren syndrome etc.And hemopoietic organ's malignant tumor also can be used as the object disease of programmed cell death inducing agent of the present invention as leukemia.Programmed cell death among the present invention brings out activity can't be confirmed when using the mice normal cell, shows thus: have described active programmed cell death inducing agent of the present invention and have high effectiveness for above-mentioned disease.In addition, programmed cell death brings out activity and can followingly measure: add medicine in cultured cell, cultivate after the stipulated time with propidium iodide etc. and carry out nuclear staining, examine under a microscope or use flow cytometer etc. to measure then.
Among the present invention, object disease as angiogenesis inhibitor, so long as need the disease of angiogenesis suppression action to get final product in the treatment, be not particularly limited, above-mentioned all malignant tumor are for example arranged, in addition also have rheumatoid arthritis, psoriasis, arthronosos deformans, senile degeneration of macula, hemangioma, hypertrophic cicatrix, periodontal disease, scleroderma etc.Angiogenesis inhibiting activity can be estimated by the following method: the fibroblast growth factor of angiogenesis etc. is brought out in interpolation in Matrigel (extracellular matrix), and it is transplanted to the subcutaneous of mice, the preparation model mice.Use extinction photometer to measure the erythrocytic hemoglobin content that soaks among the Matrigel.
Medicine of the present invention has the medicine that above-mentioned effective ingredient of the present invention and known medicine is merged preparationization with vehicle group.Can use the salt of the acceptable salt of pharmacology in the medicine of the present invention as effective ingredient.Medicine of the present invention can mix above-mentioned effective ingredient with for example known antitumor agent of other composition that has identical use with this effective ingredient, programmed cell death inducing agent or angiogenesis inhibitor, or is used in combination.
The preparation of medicine of the present invention can be undertaken by above-mentioned effective ingredient is mixed with pharmaceutically acceptable aqueous or solid carrier usually, add solvent, dispersant, emulsifying agent, buffer agent, stabilizing agent, excipient, binding agent, disintegrating agent, lubricant etc. as required, can make solid preparations such as tablet, granule, powder, powder agent, capsule, liquid preparations such as common solution, suspensoid, Emulsion.In addition, can also make and add appropriate carriers before use and make and to form aqueous goods and make dry product or other external agent.
Medicine with carrier can be according to the present invention the form of medication and the dosage form of medicine suitably select.When making the oral formulations that contains solid composite, can make tablet, pill, capsule, powder, microgranule, granule etc., for example starch, lactose, sucrose, mannose, carboxymethyl cellulose, corn starch, inorganic salt etc. can be used as carrier.During the preparation oral formulations, all right hybrid adhesive, disintegrating agent, surfactant, lubricant, fluidizer, correctives, coloring agent, essence etc.For example, when making tablet or pill, can use the thin film of sugar-coat such as sucrose, gelatin, hyprolose or gastric solubility or enteric solubility matters to coat as required.When making the oral formulations that contains fluid composition, can make the acceptable opacifiers of pharmacology, solution, suspensoid, syrup etc., for example pure water, ethanol etc. can be used as carrier.In addition, also can add auxiliary agents such as wetting agent, suspending agent, sweeting agent, flavoring agent, antiseptic etc. as required.Even effective ingredient orally give of the present invention also can be brought into play effect of sufficient, therefore consider that from administration simplicity angle making medicine for oral use is preferred form.
On the other hand, when making non-oral formulation, can be according to conventional method, with above-mentioned effective ingredient dissolving of the present invention or be suspended in distilled water for injection as diluent, normal saline, D/W, injection vegetable oil, Oleum sesami, Oleum Arachidis hypogaeae semen, soybean oil, Semen Maydis oil, propylene glycol, the Polyethylene Glycol etc., can add biocide, stabilizing agent, isotonic agent, analgesic etc. as required.Can also prepare solid composite, be dissolved in before use in sterilized water or the aseptic solvent for injection and use.
The external agent comprises solid, semi-solid or the aqueous preparation of percutaneous administration or through mucous membrane (in the oral cavity, nasal cavity) administrable.Also comprise suppository etc.For example, can make opacifiers such as Emulsion, lotion, external-use tincture, liquid preparations such as transmucosal administration liquor; Ointments such as oiliness ointment, hydrophilic ointment; Patch of percutaneous administrations such as liniment, patch, cataplasma or transmucosal administration etc.
Above-mentioned various preparation can utilize known medicinal carrying agent etc. respectively, by suitable conventional method preparation.Active constituent content in the described preparation can be considered its form of medication, medication etc., and is preferred so long as the amount that gives this effective ingredient in dosage scope described later gets final product, and is not particularly limited.Effective ingredient amount in the medicine of the present invention is about 0.1-100 weight %.
Antitumor agent of the present invention, programmed cell death inducing agent or angiogenesis inhibitor can be by the suitable route of administration administrations that adapts with dosage form.Medication is not particularly limited, can be take orally, external and injection.Injection for example can be administrations such as intravenous, intramuscular, subcutaneous, Intradermal, and the external agent for example can be with the medication administration of suppository by adapting with it.
The dosage of medicine of the present invention was suitably set according to age of the administration subject patient of its dosage form, medication, application target and this medicine, body weight symptom, cannot treat different things as the same.Usually, use when chemical compound is as effective ingredient shown in the general formula (1), the dosage of the above-mentioned effective ingredient that contains in the preparation for example per day for adults is 0.1 μ g-500mg/kg body weight, preferred 0.5 μ g-400mg/kg body weight, further preferred 1 μ g-300mg/kg body weight.Use above-mentionedly from the living compositions from the pleat umbrella of elm (drying under reduced pressure curing gained) during as effective ingredient, for example per day for adults is 0.1 μ g-1g/kg body weight, preferred 0.5 μ g-750mg/kg body weight, further preferred 1 μ g-500mg/kg body weight.Certainly, dosage changes according to various conditions, might be enough situations with the amount of lacking than above-mentioned dosage also therefore, perhaps must surpass the situation of above-mentioned scope.Administration can be in desirable dosage scope, carries out once a day or several times.Administration time is any.Medicine of the present invention can also add to arbitrarily and carry out daily picked-up in the beverage/food except that direct oral administration.
Food of the present invention is to contain chemical compound shown in the general formula (1), its derivant, the last acceptable salt of pharmacology and/or the above-mentioned food that anti-tumor activity, programmed cell death bring out activity or angiogenesis inhibiting activity that has from the living compositions from the pleat umbrella of elm, by picked-up, edible food of the present invention, particularly useful for the permanent attitude stability that keeps body.This food for example can make can indicate be used to show anti-tumor activity, programmed cell death brings out the health food (specific food for health care) of activity or the desirable effect that angiogenesis inhibiting activity brought.
The manufacture method of food of the present invention is not particularly limited.For example, mixing, modulation, processing etc. can be carried out according to general food, can make by these manufacture methods, contain the above-mentioned effective ingredient of the present invention in the gained food and get final product.
In the food of the present invention, " containing " is meant and contains, adds and/or dilute.Here, " contain " form that comprises the effective ingredient that uses among the present invention in the food that is meant, " interpolation " is meant that the effective ingredient that will use among the present invention adds the form in the raw material of food to, and " dilution " is meant the form of adding raw-food material in effective ingredient used in the present invention.
Food of the present invention is not particularly limited, and corn fabricated product (wheat flour processed goods is for example for example arranged, the starch based fabricated product, the premixing fabricated product, Noodles, the macaroni class, Bread and Pastries, bean filling class, the Semen Fagopyri Esculenti Noodles, Testa Tritici, rice noodle, vermicelli, packing rice cake etc.), oils and fats fabricated product (for example softening oils and fats, it bran sieve oil, salad oil, mayonnaise, condiment etc.), soybean processing goods (Tofu for example, bean sauce, natto etc.), meat packing goods (Petaso for example, bacon, the Petaso of compacting, sausage etc.), aquatic product (for example refrigerated flesh of fish that grinds, breaded fish stick, cylindric breaded fish stick, a flesh of fish Rhizoma Dioscoreae ball, fried Radix Dauci Sativae fish cake, fish meatball floats, muscles, fish ham, fish sausage, dry and soft fish, the fish roe processed goods, canned aquatic products, the sweet seafood etc. of cooking), milk product (raw milk for example, rare butter, clabber, butter, cheese, condensed milk, milk powder, ice cream etc.), garden stuff processing goods (cream beans class for example, jam, pickles, fruit beverage, vegetable beverage, bland etc.), snack categories (chocolate for example, Biscuits, dessert gem class, cake, rice cake dessert, rice cake class etc.), alcoholic beverage (Janpanese sake for example, Chinese wine, wine, whiskey, liquor, vodka, brandy, gin, rum, medicated beer, refrigerant alcoholic beverage, fruit wine, liqueur etc.), hobby beverage (green tea for example, black tea, oolong tea, coffee, refreshment drink, lactic acid beverage etc.), flavoring agent (soy sauce for example, sauce, vinegar, sweetener wine etc.), canned, bottled, packed food (Rice with Beef for example, the assorted set meal of pannikin, the Semen Ormosiae Hosiei glutinous rice, curried, other various deployed food), partial desiccation or reserve ration (liver pat for example, other beans, the juice of buckwheat noodles tangent plane, the condensed soup class), dried foods (instant noodles series for example, convenient curried class, instant coffee, juice powder, powder, make things convenient for bean sauce juice, deployed food, deployed beverage, deployed soup etc.), frozen food (Sukiyaki for example, Steamed Egg Custard, roasting Monopteri albi sheet, Hamburg steak, steamed dumpling with the dough gathered at the top, dumpling, various meat soups, fruit salad etc.), solid food, liquid food (for example soup etc.), fabricated product is produced in agricultural such as fragrant pungent flavoring agent class, the livestock products fabricated product, aquatic products processing is made etc.
In the food of the present invention, above-mentioned effective ingredient can contain separately or contain, add and/or dilute multiple, its content so long as be equivalent to is used to express the necessary amounts that anti-tumor activity, programmed cell death bring out activity or angiogenesis inhibiting activity and gets final product, its shape is not particularly limited, but comprises the shape thing of the oral uptake of shapes such as lamellar, graininess, capsule shape.Can add glycerol etc. in the above-mentioned effective ingredient, can make effective ingredient and obtain spissated health food.
In the food of the present invention, the amount of above-mentioned effective ingredient is not particularly limited, can suitably select according to its sense and activity expression, for example, when using chemical compound shown in the general formula (1), be preferably in the food 0.00001 weight % or more than, more preferably 0.0001-10 weight %, further preferred 0.0006-6 weight %.Use above-mentioned during from the living compositions of elm (drying under reduced pressure curing gained) from the pleat umbrella, preferably contain in the food 1 weight % or more than, more preferably 5-95 weight %, further preferred 10-90 weight %.In the food of the present invention, when wherein the mentioned component that is contained used chemical compound shown in the above-mentioned general formula (1), per day for adults can be absorbed 0.1 μ g-500mg/kg body weight, preferred 0.5 μ g-400mg/kg body weight, further preferred 1 μ g-300mg/kg body weight.Use above-mentionedly during from the living compositions from the pleat umbrella of elm (drying under reduced pressure curing gained), for example per day for adults is 0.1 μ g-1g/kg body weight, preferred 0.5 μ g-750mg/kg body weight, further preferred 1 μ g-500mg/kg body weight.
The 10th invention of the present invention also provides and needs the treatment of diseases of antitumor action, programmed cell death fall out effect or angiogenesis suppression action method, this method to comprise for what have an above-mentioned disease to be tried that chemical compound, its derivant and pharmacology go up at least a of acceptable salt shown in the above-mentioned general formula (1) that body gives effective dose.
In this description, being tried the preferred people of body, but be not limited to this, for example can be cultivated animals, pet animals etc.Cultivated animals has domestic animals such as horse, cattle, pig, sheep, goat, camel, alpaca, laboratory animals such as mice, rat, Cavia porcellus, rabbit, poultry such as chicken, duck, turkey, Ostriches.Pet animals has Canis familiaris L., cat etc.
In this description, it is above-mentioned when being tried body that effective dose is meant that at least a (effective ingredient) that chemical compound shown in the above-mentioned general formula (1), its derivant and pharmacology are gone up acceptable salt gives, with do not give this effective ingredient tried the bulk phase ratio, produce the amount of this composition of antitumor action, programmed cell death fall out effect or angiogenesis suppression action.Concrete effective dose is suitably set according to form of medication, medication, application target and age, body weight, symptom etc. of being tried body, cannot treat different things as the same, preferably identical with said medicine, above-mentioned effective ingredient amount is 0.01 μ g-500mg/kg body weight for everyone (for example adult) every day.
In the Therapeutic Method of the present invention, can be at least aly directly to give the above-mentioned body that tried, can also give with the form of said medicine or food with what chemical compound shown in the above-mentioned general formula (1) of effective dose, its derivant and pharmacology went up acceptable salt.It is unqualified to give method, for example can with said medicine similarly orally give or by the injection etc. give.
According to Therapeutic Method of the present invention, can treat the object disease of the invention described above medicine or food, the effect that for example can bring into play treatment for cancer or suppress progress, shift.
The SBS that uses as effective ingredient among the present invention does not see toxicity with 25mg/ mice/sky orally give the time.
Embodiment
Followingly describe the present invention in detail, but the present invention is not subjected to any qualification of these embodiment according to embodiment.Among the embodiment, term " tumors inhibition activity " can be used as the synonymous term use with anti-tumor activity.
Preparation example 1
To the living 5L ethyl acetate that in the sporophore dried powder of pleat umbrella M-8171, adds of 250g elm, at room temperature stirred 3 hours, filter (the No.2 filter paper that Advantec makes), remove residue, obtain 5L filtrate.With the ethyl acetate washing of 150g silica gel 60 (0.063-0.200mm) (Merck manufacturing) with about 800mL, be filled in the glass column of diameter 5cm, in silica obtained post, slowly undisturbedly inject above-mentioned filtrate, with the washing of 3L ethyl acetate, remove the not material of absorption then.Then, with 3L acetone eluting, obtain the 3L eluent, this eluent drying under reduced pressure is solidified, add 100mL 100% ethanol then, drying under reduced pressure solidifies once more, obtains the acetone elution fraction that 3.6g has removed acetone.
The result that NMR and LC-MS analyze shows: contain chemical compound shown in formula (2)-(11) in this component, its main composition composition is a chemical compound (SBS) shown in the formula (3).
Preparation example 2
Elm is living to be that elm is living from pleat umbrella K-0259 (FERM P-12981) from the pleat umbrella, in addition by the method same with preparation example 1, obtains 1.1g acetone elution fraction.NMR and LC-MS analysis result show that this component contains chemical compound shown in formula (2)-(11).
ICR mice (Japanese SCL corporate system) is a female mice of buying for 5 ages in week, uses when 6 ages in week.Sarcoma-180 (hereinafter referred to as S-180) tumor cell transplantation prepares ascites to the intraperitoneal of ICR mice, is transplanted to every 7 days again and carries out successive transfer culture in the other mice.The 7th day ascites is used the phosphate buffer centrifuge washing behind the collection successive transfer culture, is suspended in then in this buffer, carries out cell counting, is adjusted to 5 * 10
7Individual/mL.The right abdomen of this cell suspending liquid of 0.1mL being transplanted to the ICR mice is subcutaneous, measures the size of entity tumor after 7 days.With mice group, make the average of tumor size in each group, equate that every group is 10.From the living acetone elution fraction of elm from the pleat umbrella be with preparation example 1 in the components dissolved of preparation in 100% ethanol, mix with common powder feed CE-2 then, give mice.Administered dose is equal to that to be scaled elm living from pleat umbrella powder, with respect to powdered sample CE-2, with volume ratio 10% blended situation.Actual administered dose (acetone elution fraction) is about 250mg/kg/ days.Only give CE-2 in the matched group.The 4th week was measured the size of tumor after transplanting S-180.The size of tumor is to measure major diameter and minor axis, by following calculating formula volume calculated and compare.
Gross tumor volume (mm
3)=(major diameter) * (minor axis)
2/ 2
Tumor promotion calculates by following calculating formula.
Gross tumor volume * 100 of tumors inhibition activity (%)=(gross tumor volume that the gross tumor volume of matched group-the acetone elution fraction is organized)/matched group
Table 1
| Experiment numbers | Mice | Gross tumor volume (Mean +/-SE) | Tumors inhibition activity (%) |
| 1 | CE-2 (contrast) acetone elution fraction (about 250mG/kg) | 1862±542 1171±270 | 37.1 |
| 2 | CE-2 (contrast) acetone elution fraction (about 250mg/kg) | 1437±321 619±132 | 56.9 |
It is as shown in table 1 to carry out twice result of experiment.In each experiment, compare with matched group, given from elm livings in the group of pleat umbrella acetone elution fraction, the S-180 entity tumor is bred and is inhibited.
Embodiment 2
CDF
1Mice (Japanese SCL corporate system) is a female mice of buying for 6 ages in week, uses when 7 ages in week.IMC carcinoma (hereinafter referred to as IMC) tumor cell transplantation is to CDF
1The intraperitoneal of mice prepares ascites, is transplanted to every 7 days again and carries out successive transfer culture in the other mice.The 7th day ascites is used the phosphate buffer centrifuge washing behind the collection successive transfer culture, is suspended in then in this buffer, carries out cell counting, is adjusted to 5 * 10
7Individual/mL.This cell suspending liquid of 0.1mL is transplanted to CDF
1The right abdomen of mice is subcutaneous, measures the size of entity tumor after 7 days.With mice group, make the average of tumor size in each group, equate that every group is 10.From the living acetone elution fraction of elm from the pleat umbrella be with preparation example 1 in the components dissolved of preparation in 100% ethanol, mix with common powder feed CE-2 then, give mice.Administered dose is equal to that to be scaled elm living from pleat umbrella powder, with respect to powdered sample CE-2, with volume ratio 10% blended situation.Actual administered dose is about 250mg/kg/ days.Only give CE-2 in the matched group.The 4th week was measured the size of tumor after transplanting S-180.The size of tumor is to measure major diameter and minor axis, similarly to Example 1 volume calculated and tumors inhibition activity and compare.The result is as shown in table 2.
Table 2
| Mice | Gross tumor volume (Mean +/-SE) | Tumors inhibition activity (%) |
| CE-2 (contrast) acetone elution fraction (about 250mg/kg) | 3081±724 1505±246 | 51.2 |
Compare with matched group, given from elm livingly in the group of pleat umbrella acetone elution fraction, IMC entity tumor propagation is inhibited.IMC carcinoma is for CDF
1Mice is a homology, for above-mentioned homology tumor, also suppresses its propagation from elm is living from pleat umbrella acetone elution fraction.
Embodiment 3
The HL-60 cell is to use the IPM-1640 culture medium culturing that has added 10% hyclone and gentamycin with final concentration 50 μ g/mL, the cultured cell of twice gained of all subcultures.Cell with the IPMI-1640 culture medium washed twice of not adding serum, is suspended in this culture medium the counting cells number.With cell dilution is 2 * 10
6/ mL, each dispensing 1mL in 24 well culture plates.Add the acetone elution fraction of preparation in the preparation example 1, making concentration is 0.2 μ M, 2 μ M and 20 μ M, cultivates 6 hours in 37 ℃ of incubators.Carrier (1% ethanol water) is used in negative contrast, uses 1 μ g/mL actinomycin D over against shining.Cultivate after 6 hours centrifugal recovery cell.The following detection of carrying out the programmed cell death cell.Cell is at room temperature fixed 15 minutes with 4% paraformaldehyde (pH7.4), be suspended in then in 70% ethanol ,-20 ℃ of standing over night.It with phosphate buffer ((PBS) (-)) washing, is added ribonuclease A solution then, about 20 minutes of 37 ℃ of incubations.Centrifugal recovery cell adds propidium iodide solution then and dyes, and uses flow cytometer (Beckmann Coulter manufacturing) to measure the ratio of programmed cell death cell.Get a part on microscope slide, under fluorescence microscope, confirm to have taken place programmed cell death.The result as shown in Figure 1.
Embodiment 4
C57BL/6 mice (Japanese SCL corporate system) is a female mice of buying for 6 ages in week, uses when 7 ages in week.The angiogenesis evaluation of adopting Matrigel to carry out is carried out according to people's such as Passaniti (Lab.Invest.67:519-528,1992) method.Promptly, generate the factor (aFGF) to being added with 1 μ g/mL acidic fibroblast, add the acetone elution fraction of preparation in the preparation example 1 among the Matrigel of 64U/mL heparin (BectonDikinson preparation), its concentration is shown in the table 3, and this solution of 0.5mL is transplanted to the right side of mice subcutaneous abdomen.Matched group is to add the dissolving employed carrier of acetone elution fraction (1% ethanol water) to replace the acetone elution fraction.Extract Matrigel after 5 days.The Matrigel that extracts is carried out homogenate, by measuring the degree that hemoglobin content comes the comparison angiogenesis.The mensuration of hemoglobin content is used hemoglobin B-TestWako (with the pure medicine preparation of light).The result is as shown in table 3.
Table 3
| Sample | Mice (number of elements) | Hemoglobin concentration (g/mL) |
| Contrast acetone elution fraction 20 μ M | 5 4 | 1641 642 |
As mentioned above, by making the effect of acetone elution fraction, can suppress hemoglobin concentration.That is, owing to suppressed the generation of Matrigel medium vessels, so the blood flow volume that has suppressed in Matrigel, to invade.
Embodiment 5
C57BL/6 mice (Japanese SCL corporate system) is a female mice of buying for 6 ages in week, uses when 7 ages in week.The evaluation of the angiogenesis that employing Matrigel carries out is carried out similarly to Example 4.That is, it is subcutaneous that the Matrigel that has added aFGF and heparin is expelled to mice, extracts after 5 days, measures hemoglobin content.From the living acetone elution fraction of elm from the pleat umbrella be with preparation example 1 in the component lyophilization of preparation, mix with common powder feed CE-2 then, give mice.Administered dose is equal to that to be scaled elm living from pleat umbrella powder, uses the situation when mixing according to volume ratio 10% with respect to powdered sample CE-2.The about 220mg/kg/ of actual administered dose days.Matched group only gives CE-2.The result is as shown in table 4.
Table 4
| Sample | Mice (number of elements) | Hemoglobin concentration (g/mL) |
| Contrast acetone elution fraction mixes with feedstuff and gives | 5 3 | 1641 527 |
The acetone elution fraction is mixed with feedstuff when giving, still suppressed the angiogenesis among the Matrigel.Show that thus the acetone elution fraction has angiogenesis suppression action.In addition, the acetone elution fraction mixed with feedstuff visible tumor proliferation inhibitory action shows when giving, this effect is the effect that produces by the angiogenesis that suppresses tumor.
Embodiment 6
U937 cell (human lymphoma cell) is suspended in the RPMI-1640 culture medium that is added with 10% hyclone, and the counting cells number is adjusted to 5 * 10
4Individual/mL.With this suspension dispensing of each 2mL in the cell culture orifice plate.With the acetone elution fraction ethanol dilution of preparation in the preparation example 1 is 1 or 2mM, adds 20 μ L respectively, and making each final concentration is 10 or 20 μ M.Matched group only adds ethanol, at 37 ℃, 5%CO
2Cultivate under the condition, after 6 hours, after 4 hours, reclaim cell after 48 hours or after 72 hours,, count whole cell number and dead cell number with 0.3 weight % trypan blue staining.Result when only counting survivaling cell and counting is as shown in table 5.The survival results of the cell that calculates by following calculating formula is as shown in table 6.Experiment is to carry out with N=4, and the result represents meansigma methods.In addition, survival rate is calculated by following formula.
Survival rate (%)=(survivaling cell number/total cell number) * 100
Table 5 U937 survivaling cell number (is 10.0 * 10 during beginning
4Individual)
| Sample addition (μ M) | Survivaling cell number (* 10 4Individual) | |||
| Incubation time (hour) | ||||
| 6 | 24 | 48 | 72 | |
| |
12.2 9.6 10.2 | 18.3 6.2 0.5 | 37.5 4.6 0 | 86.3 3.8 0 |
The cell damage activity of table 6 couple U937 (survival rate during beginning is 100%)
| Sample addition (μ M) | Survival rate (%) | |||
| Incubation time (hour) | ||||
| 6 | 24 | 48 | 72 | |
| |
97.5 97.4 96.0 | 96.7 55.5 6.0 | 97.0 33.6 0 | 97.8 30.9 0 |
Show that more than the acetone elution fraction shows a kind of anti-tumor activity---the cell damage activity to the human lymphoma cell.
Embodiment 7
MKN45 cell (gastric carcinoma cells) is suspended in the RPMI-1640 culture medium of having added 10% hyclone, and the counting cells number is adjusted to 5 * 10
4Individual/mL.With each 2mL dispensing in the cell culture orifice plate, at 37 ℃, 5%CO
2Overnight incubation under the condition is removed culture medium, newly adds the 1mL culture medium, is 2 or 4mM with the acetone elution fraction ethanol dilution of preparation in the preparation example 1, adds 10 μ L respectively, and making each final concentration is 20 or 40 μ M.Matched group only adds ethanol.At 37 ℃, 5%CO
2Cultivate under the condition, handling with trypsin-EDTA after 6 hours, 24 hours, after 48 hours or after 72 hours, reclaim cell,, count total cell number and dead cell number, calculate survival rate similarly to Example 6 with 0.3 weight % trypan blue staining.The result who only counts survivaling cell is as shown in table 7.The result of survival rate is as shown in table 8.Experiment is carried out with N=4, and the result represents meansigma methods.
Table 7 MKN45 survivaling cell number (is 10.0 * 10 during beginning
4Individual)
| Sample addition (μ M) | Survivaling cell number (* 10 4Individual) | |||
| Incubation time (hour) | ||||
| 6 | 24 | 48 | 72 | |
| Contrast 20 40 | 11.8 7.5 6.2 | 20.7 8.2 1.8 | 43.6 6.3 0.2 | 71.5 4.6 0.1 |
The cell damage activity of table 8 couple MKN45 (survival rate during beginning is 100%)
| Sample addition (μ M) | Survival rate (%) | |||
| Incubation time (hour) | ||||
| 6 | 24 | 48 | 72 | |
| Contrast 20 40 | 96.6 95.4 82.2 | 98.0 90.0 35.4 | 97.6 81.6 2.3 | 97.4 63.5 1.0 |
As implied above, the acetone elution fraction shows a kind of anti-tumor activity-cell damage activity for gastric carcinoma cells.
Embodiment 8
S-180 cell (mice cancerous cell) or IMC cell (mice cancerous cell) use ICR mice or CDF respectively
1Mice carries out the ascites successive transfer culture, and cultured cells is transplanted subculture, gathers ascites at the 7th day then and uses., washing centrifugal with phosphate buffer are suspended among the RPMI-1640 that is added with 10% hyclone then, and the counting cells number is adjusted to 1 * 10
6Individual/mL.With its each 1mL dispensing in culture dish.With the acetone elution fraction ethanol dilution of preparation in the preparation example 1, making concentration is 1,2 or 4mM, and each adds 10 μ L, makes final concentration be respectively 10,20,40 μ M.Matched group only adds ethanol, at 37 ℃, 5%CO
2Cultivate under the condition,, use 0.2% trypan blue staining, count total cell number and dead cell number, calculate survival rate (table 9 and table 10) similarly to Example 6 after 6 hours, after 24 hours, reclaim cell after 48 hours or after 72 hours.
The cell damage activity of table 9 couple S-180 (survival rate during beginning is 98.5%)
| Sample addition (μ M) | Survival rate (%) | |||
| Incubation time (hour) | ||||
| 6 | 24 | 48 | 72 | |
| |
99.2 99.7 99.7 7.0 | 97.5 94.4 25.4 0.0 | 94.6 8.1 0.0 0.0 | 92.8 18.1 0.0 0.0 |
The cell damage activity of table 10 couple IMC (survival rate during beginning is 98.2%)
| Sample addition (μ M) | Survival rate (%) | |||
| Incubation time (hour) | ||||
| 6 | 24 | 48 | 72 | |
| |
99.0 100 92.0 44.6 | 98.7 96.7 18.3 2.3 | 95.3 71.7 2.1 1.1 | 78.5 9.3 0.9 1.8 |
Show that more than the acetone elution fraction shows a kind of anti-tumor activity for the mice cancerous cell---the cell damage activity.
Preparation example 3
Add ethyl acetate to the 300g elm is living in pleat umbrella (Lyophyllum ulmarium M-8171) sporophore powder, making total amount is 5L, stirring at room 3 hours, and extraction ethyl acetate soluble component.Filter insoluble matter by analysing, obtain the ethyl acetate soluble constituent.It is joined in silica gel 60 posts (300mL capacity), feed the 3L ethyl acetate, eluting is absorbed component not, feeds 3L acetone then, reclaims the acetone elution fraction.Add ethanol behind the concentrating under reduced pressure, drying under reduced pressure concentrates and solidifies gravimetry once more.This operation is carried out 10 times, finally by the living 35.2g acetone elution fraction that in pleat destroying angel powder, obtains of 3kg elm.This component of 500mg is crossed silica gel 60 posts, and eluting is an ethyl acetate: acetone (50: 50) carries out gradient elution to (0: 100), reclaims main component.Except that after desolvating, be dissolved in methanol: ethanol by concentrating under reduced pressure: in the mixed solvent of water (3: 4: 4), use ODS post (CAPCELLPAK C
18UG 120_ 5 μ m _ 20 * 250mm), by high performance liquid chromatography, flow velocity 10mL/ minute, detect under the condition of wavelength 210nm and prepare SBS.Eluting solvent uses the solvent of dissolving SBS.The SBS of preparation solidifies by concentrate drying, obtains 260mg purification SBS.
Embodiment 9
HL-60 cell (human leukemia cell) is suspended among the RPMI-1640 that has added 10% hyclone, and the counting cells number is adjusted to 5 * 10
5Individual/mL.With each 2mL dispensing in culture dish.With the SBS of preparation in the dimethyl sulfoxide dilution preparation example 3, making its concentration is 2mM, and each adds 20 μ L, and making final concentration is 20 μ M.Matched group only adds dimethyl sulfoxide.At 37 ℃, 5%CO
2Cultivated 6 hours, and reclaimed cell then,, washing centrifugal with phosphate buffer (PBS).In cell, add 4% paraformaldehyde, at room temperature leave standstill and fixed in 30 minutes.Remove supernatant after centrifugal, add PBS and carry out centrifugal, washing, with cell suspension in the PBS of 200 μ L.Add 500 μ L 0.05mg/mL propidium iodides, cultivated 10 minutes, measure total cell number and the cell number that programmed cell death has taken place then, calculate the ratio (table 11) of bringing out programmed cell death by following formula at 4 ℃.
Programmed cell death brings out rate (%)=(bringing out cell number/total cell number of programmed cell death) * 100
Table 11
| Sample | Programmed cell death brings out rate (%) |
| Contrast SBS (20 μ M) | 9.3 30.1 |
As mentioned above, the SBS of purification also brings out the programmed cell death in the cancerous cell.
CDF
1Mice (Japanese SCL corporate system) is a female mice of buying for 6 ages in week, uses when 7 ages in week.IMC carcinoma (hereinafter referred to as IMC) tumor cell transplantation is to CDF
1The intraperitoneal of mice prepares ascites, is transplanted to every 7 days again and carries out successive transfer culture in the other mice.Gather and transplant and the ascites of successive transfer culture after 7 days, centrifugal after suspending with phosphate buffer, precipitation is suspended in this buffer then, carry out cell counting, be adjusted to 5 * 10
7Individual/mL.This cell suspending liquid of 0.1mL is transplanted to CDF
1The right abdomen of mice is subcutaneous.Transplant the back the 7th day, and determined tumor survival and fixing, measure the size of entity tumor.With mice group, make the average of tumor size in each group, equate that every group is 12 or 11.SBS is that the SBS that will prepare in the preparation example 3 is dissolved in the ethanol, and making concentration of alcohol is 10%, with Oleum Arachidis hypogaeae semen dilution, orally give mice.Actual administered dose is about 15.6mg/kg/ days.Matched group contains 10% alcoholic acid Oleum Arachidis hypogaeae semen.In the 3rd week after the IMC tumour transplatation, the major diameter and the minor axis of mensuration tumor calculate gross tumor volume similarly to Example 1 and tumors inhibition activity compares.The result is as shown in table 12.
Table 12
| Give sample | Gross tumor volume (Mean +/-SE) | Tumors inhibition activity (%) |
| CE-2 (contrast) SBS 15.6 (mg/kg) | 992.9±179.6 534.5±92.3 | - 46.2 |
As mentioned above, in the animal groups of orally give SBS, compare with matched group, visible significant tumor proliferation suppresses.
Industrial applicability
By the present invention, can obtain cheap, contain the polyterpene that separated by edible basidiomycetes or from antitumor agent, apoptosis inhibiting agent or the AI of the living composition from the pleat umbrella of elm as active ingredient, and have the food that antitumor activity, programmed cell death bring out activity or angiogenesis inhibiting activity. Thus, the present invention is particularly particularly useful at medicine or healthy food field.
Claims (11)
1. antitumor agent, programmed cell death inducing agent or angiogenesis inhibitor is characterized in that: contain and be selected from the chemical compound shown in the following general formula (1), its derivant and their pharmacology and go up at least a as effective ingredient of acceptable salt:
[changing 1]
In the formula, m represents the integer of 1-9, and n represents the integer of 1-7, m+n=5～11, and R represents hydrogen atom or hydroxyl.
2. the antitumor agent of claim 1, programmed cell death inducing agent or angiogenesis inhibitor, wherein, the chemical compound shown in the general formula (1) is at least a chemical compound shown in following formula (2)-(11).
[changing 2]
[changing 3]
[changing 4]
[changing 5]
[changing 6]
[changing 7]
[changing 8]
[changing 9]
[changing 10]
[changing 11]
3. food, this food has anti-tumor activity, programmed cell death brings out activity or angiogenesis inhibiting activity, it is characterized in that: described food contains and is selected from the chemical compound shown in the general formula (1), its derivant and their pharmacology and goes up at least a of acceptable salt.
4. the food of claim 3, wherein, the chemical compound shown in the general formula (1) is at least a chemical compound shown in formula (2)-(11).
5. antitumor agent, programmed cell death inducing agent or angiogenesis inhibitor, it is characterized in that: it contains from the living compositions from pleat umbrella (Lyophyllum ulmarium) of elm as effective ingredient, and described compositions contains chemical compound, its derivant and their pharmacology shown in the formula of being selected from (2)-(11) and goes up at least a of acceptable salt.
6. the antitumor agent of claim 5, programmed cell death inducing agent or angiogenesis inhibitor, it is characterized in that: from the living compositions from the pleat umbrella of elm is that the living ethyl acetate extract from the pleat umbrella of elm is adsorbed in silicagel column, then the compositions that obtains of eluting.
7. food, this food has anti-tumor activity, programmed cell death brings out activity or angiogenesis inhibiting activity, it is characterized in that: described food contains from the living compositions from the pleat umbrella of elm, and described compositions contains chemical compound, its derivant and their pharmacology shown in the formula of being selected from (2)-(11) and goes up at least a of acceptable salt.
8. the food of claim 7, it is characterized in that: from the living compositions from the pleat umbrella of elm is that the living ethyl acetate extract from the pleat umbrella of elm is adsorbed in silicagel column, then the compositions that obtains of eluting.
9. from the living compositions of elm from the pleat umbrella, said composition contains chemical compound, its derivant and their pharmacology shown in the formula of being selected from (2)-(11) and goes up at least a of acceptable salt, and said composition is that the living ethyl acetate extract from the pleat umbrella of elm is adsorbed in silicagel column, and eluting obtains then.
10. treatment of diseases method, described disease must take antitumor action, programmed cell death fall out effect or angiogenesis suppression action to treat, and this method comprises: at least a body that tried of suffering from this disease of chemical compound, its derivant and their pharmacology shown in the above-mentioned general formula of being selected from of effective dose (1) being gone up acceptable salt.
11. be selected from least a application in preparation antitumor agent, programmed cell death inducing agent or angiogenesis inhibitor that the chemical compound shown in the above-mentioned general formula (1), its derivant and their pharmacology go up acceptable salt.
Applications Claiming Priority (4)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| JP162641/2004 | 2004-05-31 | ||
| JP2004162641 | 2004-05-31 | ||
| JP243836/2004 | 2004-08-24 | ||
| JP292536/2004 | 2004-10-05 |
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| Publication Number | Publication Date |
|---|---|
| CN101001617A true CN101001617A (en) | 2007-07-18 |
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ID=38693298
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|---|---|---|---|
| CN 200580025889 Pending CN101001617A (en) | 2004-05-31 | 2005-05-30 | Antitumor agent |
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| CN (1) | CN101001617A (en) |
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2005
- 2005-05-30 CN CN 200580025889 patent/CN101001617A/en active Pending
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