CN104447266A - Compound used as antitumor drug and food - Google Patents

Abstract

The intention provides a compound used as an antitumor drug or tumor prevention and heath care food. The structural formula is shown in the specification.

Description

The compound of a kind of antitumor drug and food

Technical field

The present invention relates to novel cpd, antineoplastic agent and there are the pharmaceuticals of antitumor action, food or makeup.

Background technology

Antineoplastic agent carcinostatic agent carcinostatic agent is select optionally to act on tumour cell and the preparation less to normal cytotoxicity substantially, but also have the problem on a lot of use to be resolved not yet, the appearance of the fatal side effects such as the leukopenia such as caused by bone marrow depression, thrombopenia, Neutrophilic granulocytopenia, the appearance of the side effect of the Digestive tract obstacles such as nausea and vomiting, and the appearance of the side effect such as alopecia, drug-fast appearance, and because the medicament being difficult to oral administration, route of administration is restricted.Therefore, expect to develop and optionally act on tumour cell and, route of administration few to normal cytotoxicity limits few antineoplastic agent carcinostatic agent carcinostatic agent.

As the antineoplastic agent obtained from natural goods, there will be a known a large amount of compounds, such as, have the report that ametycin, camptothecine, triterpenoid etc. are a large amount of.In addition, as the antineoplastic agent taking natural goods as lead compound, there will be a known D51-7059, Taxane derivative etc.As phone predicts, the content of the antineoplastic agent of naphthoquinones system that is relevant to this compound or that be correlated with the biosynthesizing system of this compound, decomposing solution, has the report about the kurarinone from kuh-seng.In addition, as the synthetic compound relevant to naphthoquinones system antineoplastic agent, there will be a known the antineoplastic agent relevant to furans naphthoquinone derivatives.

Summary of the invention

Novel cpd of the present invention, can be obtained kuh-seng by extraction purification operation as raw material, also can obtain in other plant materials.In addition, also can use by synthesizing the novel cpd obtained.But also kuh-seng can be used, from containing extracting the extract obtained, thick purified or the dry thing of plant materials, the slurry of plant materials other plant materials of novel cpd.

From the plant materialss such as kuh-seng during extraction purification novel cpd of the present invention, any extraction purification operation of usual industrial use can be used.After as the leaf, stem, root, flower etc. of raw material, the phase gathers in due course, directly or implement the drying process of usual air seasoning etc., extraction raw material is made.

That is: by after raw material pulverizing or chopping, extract with solvent, as Extraction solvent, can by water, the alcohols such as ethanol, methyl alcohol, Virahol, the ketone such as acetone, methyl ethyl ketone, the ester such as methyl acetate, ethyl acetate class, the oil loving solvent such as hexane, chloroform is used alone or is formed mixed solvent and uses.Extracting temperature is generally 0 ~ 100 DEG C, is preferably 5 ~ 50 DEG C.Extraction time is 1 hour ~ about 10 days, and quantity of solvent is that on average every part of dried feed is generally 1 ~ 30 times of weight, is preferably 5 ~ 10 times of weight.Extraction operation, by stirring, is also undertaken by dipping placement.Extraction operation repeatedly can carry out 2 ~ 3 times.By filtering or centrifugation and removing in the extracting solution after insoluble sludge or the method for each novel cpd of purifying from the squeezeding juice of plant from the crude extract obtained with aforesaid operations, as long as known crude drug separation purification method, any method can be adopted, but be preferably used alone or in combination two-phase solvent apportion design, counter-current distribution method, column chromatography, preparative high performance liquid chromatography etc.Such as two-phase solvent apportion design, can exemplify by distributing in normal hexane, chloroform, methyl ethyl ketone, ethyl acetate, methyl acetate equal solvent and water, thus target compound is recovered to the method etc. in solvent phase from said extracted liquid.As column chromatography, ion exchange column chromatography method, use positive or the reverse phase silica gel modification dextrane gel as the method for carrier, the Adsorption column chro-matography using DIAION HP-20 etc., use SephadexLH-20 etc. can be exemplified as the gel filtration method etc. of carrier, these methods can be used alone or in combination or use repeatedly.As preparative high performance liquid chromatography, the method for the reversed-phase column using octadecyl silicon etc. can be exemplified, use the method etc. of the normal phase column of silica gel etc.

As the route of administration of antineoplastic agent of the present invention, be not particularly limited, such as, can exemplify administration in the intestines such as oral administration drop rectum with drug, the mucosa delivery of nose administration etc., the drug administration by injection etc. of intravenous administration subcutaneous administration etc.As the formulation of antineoplastic agent of the present invention, form that is any and the matched preparation of medication can be taked, such as can exemplify tablet, powder, granula subtilis, granule, capsule, powder, pill, containing solid preparations such as tablets, the liquid preparations such as solution, suspension, emulsion, syrup, injection, gel preparation etc.Can by directly administrations such as the sterling of each novel cpd, purified, thick purifieds, also can with the excipient pharmacologically allowed together administration.As excipient, as long as monose, disaccharides, polyose, inorganic salts, grease, distilled water etc. are as the usual spendable excipient of preparation, any excipient can be used.When carrying out formulation, also can use the additives such as tackiness agent, lubricant, dispersion agent, suspension agent, emulsifying agent, thinner, buffer reagent, antioxidant, bacterial inhibitor.

Effective dosage of each novel cpd, different according to the difference at the symptom of route of administration, formulation, disease, the age of object etc., be generally and be 0.1 ~ 1000mg, be preferably 0.5 ~ 300mg, more preferably 1 ~ 100mg every day for each person.The content of each novel cpd in oral antineoplastic agent of the present invention, can set and the active constituent content in the optimal preparation of each administration form according to the data of the effective dosage of the form of preparation as the dosage of preparation.

As the form of food, form kuh-seng and the dry thing of plant materials containing each novel cpd being made tea can be exemplified, or be combined with the food etc. of the sterling of each novel cpd, the partial purification product of this novel cpd, the crude extract from containing this novel cpd extracted the plant of this novel cpd, the plant materials slurry containing this novel cpd, the dry thing of the plant materials containing this novel cpd.

As tea, can separately or used in combination with other tea raw materials.As other tea raw material, as long as green tea, oolong tea, Leaf of Assam Tea, black tea, simmer tea, brown rice tea, Eucommia Tea, persimmon-leaf tea, mulberry tea etc. usually used as the edible raw material of tea, any tea raw material can be used.

When obtaining plant milk extract, as long as by hot water extraction, by spendable method in ethanol, aqueous ethanol extraction etc. usually food extraction, any method can be used.From plant materials, crude extract, the partial purification product of each novel cpd are obtained by well-established law.

As the form with the food of antitumor action of the present invention, except tea, as long as nourishing drink, jelly, biscuit, tablet, pill, soft capsule, hard capsule, powder, granula subtilis, granule etc., usually used as the available form of food, can use any form.As auxiliary material, also the additives such as excipient, tackiness agent, lubricant, dispersion agent, suspension agent, emulsifying agent, thinner, buffer reagent, antioxidant, bacterial inhibitor can be used.

The present invention has effective intake of each novel cpd of the food of antitumor action, different according to the difference at picked-up form, the state of health of object, the age of object etc., but being generally every day is for each person 0.001 ~ 100mg, be preferably 0.01 ~ 10mg, more preferably 0.1 ~ 1mg.

The present invention has the content of each novel cpd in the food of antitumor action, different according to the difference of food form, but is generally 0.0001 ~ 1wt%, is preferably 0.001 ~ 0.5wt%, is more preferably 0.01 ~ 0.1wt%.

As the example of the form of external application pharmaceuticals of the present invention and makeup, be not particularly limited.

As the form of external application pharmaceuticals, such as, can exemplify ointment, creme, paste, adhesive tape agent, external preparation etc.Pharmaceuticals of the present invention, on the basis of each novel cpd, can contain other medical components as required, in addition, also can use the additives such as tackiness agent, dispersion agent, suspension agent, emulsifying agent, thinner, buffer reagent, antioxidant, bacterial inhibitor.

As the form of makeup, astringent, beauty liquid, emulsion, breast frost, gel, facial mask, beauty treatment muffin, cleansing milk, baths etc. can be used as the spendable any form of external preparation cosmetic formulations.In above-mentioned makeup, on the basis of the required compositions such as the partial purification product of the sterling of each novel cpd, this novel cpd, the crude extract of this novel cpd extracted from plant or the plant materials containing this novel cpd, also can as required containing the composition that can coordinate in cosmetic formulations.As gradation composition, such as, can exemplify solid oil, lard, liquid oils, low molecule wetting Agent for Printing Inks, polymer wetting Agent for Printing Inks, fat-soluble wetting Agent for Printing Inks, softener, tensio-active agent, sanitas, antioxidant, pH adjusting agent, ethanol, water etc.

Effective dosage of each novel cpd used time outside, according to the symptom of object, the difference at the age of object and different, but to be generally every day be for each person 0.001 ~ 100mg, is preferably 0.01 ~ 10mg, more preferably 0.1 ~ 1mg.

The content of each novel cpd in antineoplastic agent of the present invention, pharmaceuticals, makeup, separately or be generally 0.0001 ~ 1wt% as during mixture, is preferably 0.001 ~ 0.5wt%, is more preferably 0.01 ~ 0.1wt%.

Embodiment

Below by embodiment, the invention will be further described.The method described in the embodiment of the present invention that it should be understood that is only used for the present invention is described, instead of limitation of the present invention, under concept thereof of the present invention, all belong to the scope of protection of present invention to the simple modifications of preparation method of the present invention.The all raw materials used in embodiment and solvent are all purchased from Sigma Biochemical and Organic Compounds for Research and Diagnostic Clinical Reagents company.

Embodiment 1: the preparation of formula I

At room temperature, with 90% (v/v) ethanol of 25L, the extraction that each 24 hours amount to 3 times is carried out to kuh-seng leaf (5Kg), concentrate after these are combined, obtain dry substance (407g).Then, after making its dissolving that suspends in 90% (v/v) methyl alcohol of 7L, after carrying out 3 sub-distribution with the hexane of equivalent, take out 90% (v/v) methanol phase and carry out concentrating under reduced pressure.In this concentrating under reduced pressure thing, add pure water to 5L, move in separating funnel and carry out 3 two-phase solvents distribution with chloroform.Then the chloroform obtained by this operation is combined mutually, obtain dry substance 69.3g.By wherein 69.0g supply using the silica gel column chromatography (80mm φ × 150mm, Kanto Kagaku K. K. system) of hexane/ethyl acetate as eluting solvent.That is: after successively post being washed by the eluting solvent hexane/ethyl acetate (9: 1) of 3BV and the eluting solvent hexane/ethyl acetate (8: 2) of 2BV, use eluting solvent hexane/ethyl acetate (7: the 3) wash-out of 2BV again, obtain B component (dry substance 5.66g).Then, for B component, methyl alcohol is implemented Sephadex LH-20 column chromatography (20mm φ × 200mm, Pharmacia Inc.) as eluting solvent.That is:, after being washed by post with the methyl alcohol of 2.5BV, by the methanol-eluted fractions of 1BV, B component-2 (dry substance 480mg) is obtained.Then, by B component-2 (dry substance 480mg) supply using Flash ODS column chromatography (20mm φ × 150mm, the wild village chemical company system of water/methyl alcohol as eluting solvent, that is: after Flash ODS post being washed with 50% (v/v) methyl alcohol of 180ml, after using 60% (v/v) methyl alcohol, 70% (v/v) methyl alcohol, 80% (v/v) methanol wash step by step successively, use 90% (v/v) methanol-eluted fractions again, obtain the component X (dry substance 143mg) containing target novel cpd D, E.And then, with silica gel column chromatography (20mm φ × 150mm, Kanto Kagaku K. K.'s system), component X (dry substance 143mg) is distinguished.That is: after silicagel column being washed by the eluting solvent hexane/ethyl acetate (8: 2) of 2BV, with eluting solvent hexane/ethyl acetate (7: the 3) wash-out of 1BV, obtain B component-2-1 (dry substance 32.3mg) and B component-2-2 (dry substance 18.8mg).By preparative high performance liquid chromatography (ODS post, 20mm φ × 250mm, chemical company of wild village system, moving phase: 58% (v/v) acetonitrile, detect: 280nm UV detector) carry out purifying, from B component-2-1 (dry substance 32.3mg), obtain formula I (dry substance 5.0mg).

¹H NMR(CDCl ₃，500MHz)：δ0.98(H-12and H-13，6H，s)、1.98(H-8’，3H，s)、2.24(H-7’，3H，s)、2.65(H-9，2H，s)、3.90(H-11，2H，s)、6.21(H-5’，1H，d)、7.08(H-3’，1H，d)、7.25(H-4’，1H，dd)、7.60(H-7，1H，t)、7.67(H-6，1H，t)、7.99(H-8，1H，d)、8.03(H-5，1H，d).

Embodiment 2: anti-tumor activity is tested

Study on antitumor effect is undertaken by the proliferation inhibition activity measured for the melanoma cell strain HMV-II from human body.

Use the Ham F12 substratum containing 10% (v/v) foetal calf serum, with 1 × 10 on 96 hole culture plates (NUNC) ⁴the cell concn inoculation of cells/90 μ l from the melanoma cell strain HMV-II of human body, 37 DEG C, under 5% carbon dioxide exists, carry out cultivating for 24 hours.Cultivate after 24 hours, adding formula I of the present invention (is dissolved in DMSO, final DMSO concentration=0.1% (v/v) in substratum, in addition, by cultured cells strain on the substratum of DMSO only adding 1/1000 volume as a control group), so 37 DEG C, 5% carbon dioxide exist under cultivate 24 hours.The mensuration of relative growth rate uses tetrazolium bromide (MTT) [3-(4,5-dimethylthiazole-2-base)-2,5-phenylbenzene bromination tetrazole Nacalai Tesque] [(3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazol ium bromide)] method carry out [with reference to Co., Ltd. Tokyo chemistry with people's distribution, new biochemical experiment lecture 12 molecular immunology I immune cells factor 358-359 page].That is: add after above-mentioned each novel cpd cultivates 24 hours, at 96 hole culture plate (0.33cm ²/ well) each hole nutrient solution 90 μ l in add the MTT solution { 5mg/ml of 10 μ l; After dissolving in without calcium magnesium PBS [Du Shi phosphoric acid buffer (Dulbecco ' s Phosphate-Buffered Saline)] solution, solution after film filter (0.22 μm) filters }, carrying out vibration makes it even, 37 DEG C, 5% carbon dioxide cultivates 4 hours under existing.Cultivate after 4 hours in each hole, add 10% (w/v) SDS-50% (v/v) N, dinethylformamide-0.005N hydrochloric acid soln 100 μ l, 37 DEG C, 5% carbon dioxide leaves standstill after 18 hours under existing, use immune detector (immuno reader) (Dainippon Pharmaceutical Co., Ltd's system), with 750nm in contrast, measure the absorbancy at 590nm place, as the index of relative growth rate.Formula I of the present invention is under the concentration of 10 μ g/ml, and appreciation rate is 84%.The excellent proliferation inhibition activity of formula I to the melanoma cell strain (HMV-II) from human body can be confirmed.

Claims (3)

1. a compound, is characterized in that it is with shown in following structural formula:

I。

2. an antineoplastic composition, is characterized in that containing the compound shown in formula I and pharmaceutically acceptable auxiliary agent.

3. a protective foods, is characterized in that containing the compound shown in formula I.