New cyclohexenone compounds of antrodia camphorata and their application |
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IPC classes for russian patent New cyclohexenone compounds of antrodia camphorata and their application (RU 2422431):
    
![Pyrrolo[2,3-b]pyridine derivative as protein kinase inhibitors](http://img.russianpatents.com/img_data/1078/10788328-s.gif) Pyrrolo[2,3-b]pyridine derivative as protein kinase inhibitors / 2418800
Invention describes a compound of structural formula IIIm:
 Novel cyclohexane derivative, prodrug and salt thereof and diabetes therapeutic agent containing said compounds / 2394015
Present invention relates to novel cyclohexane derivatives of formula (I) or their pharmaceutically acceptable salts having inhibitory effect on Na+-glucose cotranspoter (SGLT2), as well as to pharmaceutical compositions based on the said compounds and their use in preventing or treating diabetes, diabetic complications caused by hyperglycaemia or obesity.
![Oligo(4[2-(trimethoxysilanyl)ethyl]-2-cyclohexyloxy)-hydroxymethoxyphenyl-1-pentafluorophenylpropenone as precursor for modifying silica particles in sol-gel process, method of producing said compound and method of modifying silica particles with precursor sol-gel process](http://img.russianpatents.com/img_data/1048/10484438-s.gif) Oligo(4[2-(trimethoxysilanyl)ethyl]-2-cyclohexyloxy)-hydroxymethoxyphenyl-1-pentafluorophenylpropenone as precursor for modifying silica particles in sol-gel process, method of producing said compound and method of modifying silica particles with precursor sol-gel process / 2383551
Proposed is a precursor in form of oligo(4-[2-(trimethoxysilanyl)ethyl]-2-cyclohexyloxy)-hydroxymethoxyphenyl-1-pentafluorophenylpropenone, method of producing said compound through cationic copolymerisation with opening of the epoxide ring of the organic compound having an epoxide group and a trialkoxysilyl group, and an epoxy-derivative of an organic functional molecule meant for modifying the surface or particles of silica and a method of modifying silica particles in a sol-gel process. Also proposed is a method of modifying silica particles in a sol-gel process using the described precursor. Silica particles modified by the said precursor can be used to indicate presence of amino-containing compounds, particularly phenylhydrazine.
 Naphthalene derivatives / 2354646
Invention refers to new compounds of formula (I) and to its pharmaceutically acceptable acid-additive salts. The compounds under the present invention are active to bind cannabinoid (CB) receptor. In general formula (I)
 Novel trifunctional photoinitiators / 2348652
Invention concerns new photoinitiators, method of their obtainment, compositions hardening with irradiation, and application of those compositions in coating preparation. Invention claims photoinitiators of formulae I
 Method for preparing4,4-dimethoxy-2,3,5-trichlorocyclopent-2-en-1-one / 2320637
Invention relates to a method for synthesis of 4,4-dimethoxy-2,3,5-trichlorocyclopent-2-en-1-one from hexachlorocyclopentadiene. Method involves carrying out two successive steps - interaction of hexachlorocyclopentadiene with potassium hydroxide in methanol medium in the ratio = 1:(8-10) at temperature 20-22°C for 24 h in a single reactor and acid hydrolysis of formed intermediate substance with 50% aqueous sulfuric acid without its isolation. The end substance is isolated by vacuum distillation with the yield 75-80% and purity 96-98%. Invention allows excluding the high cost and difficultly available reagents, simplifying the process and increasing yield of the end substance.
 Multi-purpose plant for rectification cleaning of solvents belonging to main classes of organic solvents and method of rectification cleaning of acetone, isopropyl alcohol, benzene, toluene, n-butanol, isobutanol, ethyl acetate, n-butyl acetate and cyclohexane / 2264840
Proposed multi-purpose plant includes still and batch-operated packed rectifying column mounted on it and brought in communication with it; top of said column on side of vapor outlet is communicated with dephlegmator which is brought in communication with top of rectifying column on side of condensate outlet and with rectification production collecting reservoir; proposed plant is provided with two measuring reservoirs connected at their lower parts to starting solvent reservoir located above them and to still; at their upper parts they are connected to vacuum pump and to nitrogen source; measuring reservoirs are located above still for filling it by gravity; volume of measuring reservoirs ranges from 60 to 75% of still volume; dephlegmator is made in form of cylindrical water-cooled heat exchanger mounted at angle of 10 to 20° relative to level; in its lower part, dephlegmator is provided with bypass unit with combined vapor inlet and condensate outlet; bypass unit is used for connecting the dephlegmator to top of rectifying column on side of solvent vapor outlet and to point of introducing the phlegm to rectifying column and to reservoirs collecting predistillate and rectification products; besides that, dephlegmator is connected with separator for separation of water from condensate (Florence flask) which may be disconnected when necessary; water receiver is located below it and is brought into communication with it; stillage residue receiver located above still is connected to its lower part; stillage residue receiver is connected by its lower part to stillage residue utilization reservoirs; at the top it is connected to vacuum pump and to nitrogen source; predistillate and reaction qualification products are accumulated in separate reservoirs connected to column condensate outlet; plant is provided with trap for receiving non-condensed vapor of rectification products which is connected with water receiver, dephlegmator and water cooler; trap is connected to predistillate collecting reservoir; rectifying column consists of four drums of similar height hermetically interconnected together; their diameter ranges from 0.07 to 0.08 of rectifying column height; height of rectifying column ranges from 3800 to 4200 mm; volume of predistillate and stillage residue collecting reservoir is 20-30% of volume of still; volume of starting solvent reservoir is 200-300% of volume of still; rectifying column, starting solvent reservoir and rectification product collecting reservoir are made from stainless steel.
 Method for preparing of 5- and/or 6-substituted 2-hydroxybenzoic acid esters / 2245325
Invention relates to the improved method for preparing compound of the formula (I):
 Methods of obtaining intermediates / 2224740
 The method of acylation of aromatic compounds / 2223938
The invention relates to a method for the acylation of aromatic compounds of the formula I
 < / BR>in which a represents the residue of the benzene cycle; radical or radicals R are identical or different and denote one of the following groups: alkyl, linear or branched, having 1-6 carbon atoms, preferably 1-4 carbon atoms, such as methyl, ethyl, propyl, isopropyl, butyl, isobutyl, sec.butyl, tert.butyl, alkoxy, linear or branched, having 1-6 carbon atoms, preferably 1-4 carbon atoms, such as methoxy, ethoxy, propoxy, isopropoxy, butoxy, a radical of the formula-R1-X, in which R1mean valence bond; X is a halogen atom, preferably a chlorine atom, bromine or fluorine, n is a number less than or equal to 4, preferably 0 or 1, by reaction of the compounds with allermuir agent of formula II  < / BR>in which X' represents a chlorine atom and R3means alkyl, linear or branched, having from 1 to 12 carbon atoms, preferably 1-4 carbon atoms, in the presence of a zeolite catalyst, characterized in that carry out the mixing in any way aromaterapiey layer of catalyst, and recycle the reaction mixture was released from the catalytic layer, through the catalytic layer as many times as necessary to obtain the desired degree of conversion of the aromatic compounds
Method of extracting vanilin, ethylvanilin, isovanilin and orto-vanilin from water solutions / 2352553
Method of extracting vanillin, ethylvanillin, iso-vanillin and ortho-vanillin from water solutions is characterised by the following: preliminarily to water solutions of vanillin, ethylvanillin, iso-vanillin and ortho-vanillin added are; ammonium sulphate, until its content in solution is 42-43% to solution weight, and preliminary prepared mixture of solvents, consisting of 22-23 wt % acetone and 78-77 wt % diacetone alcohol, then extraction is carried out with molar ratio of water and organic phases 10:1, and degree of vanillins extraction (R,%) is calculated by formula: R=D-100/(D+r), where D is coefficient of vanillins distribution between solvent mixture and water-salt solution, r is ratio of equiweight volumes of water and organic phases.
 Method of cleaning acrolein / 2315744
Proposed method consists in continuous cleaning of acrolein. Aqueous solution of acrolein free from difficult-to-condense gas is fed to distilling column equipped with at least one evaporator mounted in its base and at least one condenser mounted in its upper part. Mixture mainly containing water is discharged from distilling column base. Mixture mainly containing acrolein and water is discharged from upper part of distilling column. Mixture discharged from upper part of distilling column is cooled in condenser to temperature at which aqueous condensate may be obtained in addition to considerable amount of acrolein-rich gaseous mixture.
 The allocation method chlorella / 2160247
The invention relates to chemical technology and relates to a method of allocation Chlorella products from the chlorination of ethanol
Extraction of vanillin / 2065434
 Acetamide compounds as fungicides / 2396268
Invention describes compounds of formula (1)
 Method for synthesis of 3-(methylthio)propanal and 2-hydroxy-4(methylthio)butane nitrile / 2383531
Invention relates to a method for synthesis if 3-(methylthio)propanal and 2-hydroxy-4-(methylthio)butane nitrile through a catalytic reaction for respectively bonding methylmercaptan to acrolein and hydrocyanic acid to 3-(methylthio)propanal. In the said methods, the catalyst is heterogeneous, does not dissolve in the reaction medium and has formula (I)
Method of producing 3-methylthiopropanal / 2382767
Method involves a stage where acrolein and methylmercaptan are fed into a reaction system simultaneously or successively relative an acid compound or base compound for reaction of acrolein and methylmercaptan, where the base compound is used in amount of approximately 0.3 mol or less per mole of the acid compound, where the acid compound is a carboxylic acid and the base compound is a nitrogen-containing organic base.
 Method for preparation of 3-(alkylthio)propanal / 2367652
Invention refers to the method for preparation of 3-(alkylthio)propanal of general formula I
 Method of obtaining 3-methyltiopropanal / 2336266
Method of obtaining 3-methyltiopropanal includes interaction of methylmercaptane with acrolein in presence of catalyst, containing organic base and organic acid, organic acid representing N-alkylmorpholin.
 Compounds for controlled releasing active molecules / 2296118
Invention relates to an aromatizing composition containing at least compound of the formula (I):
Method for preparing γ-ketosulfide concentrates / 2278110
Invention relates to a method for preparing concentrates of γ-ketosulfides. Method for preparing concentrates of γ-ketosulfides is based on the alkylthiomethylation reaction of ketones with a mixture of formaldehyde and methane gas odorant containing mercaptanes and disulfides in an aqueous-alkaline medium at temperature 20 ± 5°C. Proposed method provides enhancing the yield of the end product. Concentrates of γ-ketosulfides can be used as foaming agents-collecting agents in flotation of copper-zinc and gold-containing ores or adsorbents of sulfur dioxide.
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FIELD: medicine, pharmaceutics. SUBSTANCE: present invention refers to new compounds of formula EFFECT: production of the new compounds inhibiting tumour cell growth. 31 cl, 7 tbl, 8 ex
The technical field to which the invention relates. The present invention relates to a new connection, in particular to extract, isolated and purified from Antrodia camphorata and its application in the suppression of tumor growth. The level of technology Antrodia camphorata (Niu Chang MS), also known as Chang-JI", "Niu-Chang-Ku", "Ed Chang, Ed Chang-Chi, Chung-Ku, camphor chamber mushroom and so on, is an endemic species in Taiwan, growing on the inside of the rotten core of the wood walls of the thicket cinnamon Cinnamomum kanehirae hay on the altitude from 450 m to 2000 m in the mountains of Taiwan. Fruiting body of Antrodia camphorata is growing inside the tree trunk. Cinnamomum kanehirae Hey distributed mainly in mountain areas, Tao-yuan, Nan-Tou and included in the list of rare and valuable species due to the small numbers, and excessive deforestation is illegal. Thus, Antrodia camphorata in nature is also rare. Since the growth rate of natural Antrodia camphorata is extremely low, and the growing season runs from June to October, the price of Antrodia camphorata is very high. Fruiting bodies of Antrodia camphorata are perennial, escheresque, cork or wood, with a variety of external shapes, such as plates, bells, hooves or tower. They are flat on the wood surface at the beginning of growth. Then the edges on the front line looking is are stated, wrapping in the form of plates or stalactites. The upper surface of Antrodia camphorate are shiny, brown or dark brown, with subtle folds flat and blunt. The lower sides are orange-red or partly yellow with pores on the entire surface. In addition, Antrodia camphorata emits a strong smell of sassafras (camphor aroma), becomes pale yellow-brown after drying in the sun, and has a strong bitter taste. In traditional Taiwanese medicine Antrodia camphorata is typically used to detoxify, protect the liver, as anti-cancer tool. Antrodia camphorata as usual edible and medical mushrooms, rich in numerous nutrients, including polysaccharides (such as p-Glucosan), triterpenoids, superoxide dismutase (SOD), adenosine, proteins (immunoglobulins), vitamins (such as vitamin b, nicotinic acid), minerals (such as calcium, phosphorus and germanium, etc.), nucleic acid, agglutinin, amino acids, steroids, lignins and stabilizers blood pressure (such as antrodia acid) and the like. I believe that these bioactive ingredients have a favorable effects, such as antitumor, strengthening immunity, protivoallergichesky, inhibition of agglutination of platelets, antiviral, protivoraketa the territorial, antihypertensive, lowering the level of glucose in the blood, lower cholesterol, protect the liver, and the like. Triterpenoids are the most studied component of among the numerous compositions of Antrodia camphorate. Triterpenoids are the common name for natural compounds containing 30 carbon atoms with or Pyh exactlyonce structures. The bitter taste of Antrodia camphorata is caused by the component of the triterpenoids. Three new triterpenoids ergotaminovogo type (antzen And antzen, antzen C) were selected Chemg et al. from the fruiting bodies of Antrodia camphorata (Chemg, I. H., and Chiang, H. C. 1995. Three new triterpenoids from Antrodia cinnamomea. J. Nat. Prod. 58:365-371). Three new compounds, named gancheva acid And gancheva acid and gancheva acid were extracted from the fruiting bodies of Antrodia camphorata ethanol Chen et al. (Chen, S., and Yang, S. W. 1995. New steroid acids from Antrodia cinnamomea, a fungus parasitic on Cinnamomum micranthum. J. Nat. Prod. 58:1655-1661). In addition, Chemg et al. also found in the fruiting bodies of Antrodia camphorata other three new triterpenoids, which of sesquiterpenes the lactone and the two compounds derived from biphenyl, 4,7-dimethoxy-5-methyl-1,3-benzodioxole and 2,2',5,5'-tetrametoksi-3,4,3',4'-bi-methylendioxy-6,6'-dimethylbiphenyl (Chiang, H. C., Wu, D. R., Chemg, I. W., and Ueng, S. 1995. And sesquiterpene lactone, phenyl and biphenyl compounds from Antrodia cinnamomea. Phytochemistry. 39:613-616). In 1996 four new triterpenoids ergotaminovogo type (attiny E and F and m is Teal, uncinate G and H) were selected Chemg et al. using the same analytical methods (Chemg, I. H., Wu, D. P., and Chiang, H. S. Triteroenoids from Antrodia cinnamomea. Phytochemistry. 41:263-267). Two related to ergostane steroid gancheva acids D and E along with three of lunesta-related triterpenes, 15 alpha-acetyl-dihydroalprenolol acid, dehydroabietic acid and dihydroalprenolol acid were isolated Yang et al. (Yang, S. W., Shen, Y. C., and Chen, S.N. 1996. Steroids and triterpenoids of Antrodia cinnamomea is a fungus parasitic on Cinnamomum micranthum. Phytochemistry. 41:1389-1392). The search for the exact active ingredients with anti-tumor effect are still at an experimental stage and need to be clarified, although it was reported antitumor effects of Antrodia camphorata (such as in the above links). If you find the exact antitumor composition, it will contribute to the beneficial effects of cancer treatment. Disclosure of inventions To identify anticancer compounds from extracts of Antrodia camphorata, the present invention was isolated and purified compound of formula (1),
in which X and Y can be oxygen or sulfur, R1, R2and R3each are a hydrogen atom, stands or (CH2)m-CH3and m=1-12; n=1-12. The preferred compound of General formula (1) is 4-hydroxy-2,3-dimethoxy-6-methyl-5(3,7,11-trimethyl-dodeca-2,6,10-trienyl)-cyclo is CEN-2-Aenon as shown in the formula (2), with the molecular formula C24H38O4in the form of a pale yellow powder, with a molecular weight of 390.
Compounds with structure of formula (1) and formula (2) is purified by aqueous extraction or extraction with an organic solvent Antrodia camphorata. Organic solvents include without limitation alcohols, such as methanol, ethanol or propanol, esters such as ethyl acetate, alkanes such as hexane, or alkyl halides, such as Harmattan, charatan. Among them, preferred is an alcohol, particularly preferred is ethanol. Compounds that can be used in accordance with the invention, can be suppressed the growth of tumor cells that can be further applied to a medical composition for the treatment of cancer and enhance therapeutic effects. Compounds of the invention can be applied for a number of cancer cells, including breast cancer, liver cancer and prostate cancer, which slows down the growth of cancer cells, with subsequent suppression of proliferation of cancer cells and reducing the risk of tumor development. Thus, they can be used in the treatment of cancer, such as breast cancer, liver cancer, prostate cancer, and the like. On the other hand, the compounds of formula (1) and/or formula (2) in the invention can be included in the medical the e compositions for the treatment of breast cancer, liver cancer and prostate cancer, to inhibit the growth of tumor cells. Medical compositions include not only the compounds of formula (1) and/or formula (2), but also pharmaceutically suitable carriers. Carriers include, without limitation fillers such as water, sealants, such as sucrose or starch, binding agents such as derivatives of cellulose, diluents, disintegrant, amplifiers absorption or sweeteners. The pharmaceutical composition of the present invention can be produced by mixing the compounds of formula (1) and/or (2) at least one of the media using conventional methods known in the pharmaceutical art and can be formulated without limitation in the form of powder, tablets, capsules, pills, granules or other liquid formula. In addition, since the compounds of the present invention at the same time have antioxidant activity, they can be perfect additions to a healthy diet, diets and drinks, health products and cosmetics, and are favorable for human health because of their talents to the prevention of cardiovascular disease or mutation of cells. The present invention is further explained in the following examples and illustrations of embodiments. These examples, however, should not be considered as limiting the volume of the m invention, and it is assumed that specialists in this field of technology can be easily made modifications within the invention and scope of the attached formula. The implementation of the invention Mycelium, fruiting bodies, or their mixture from Antrodia camphorata was first extracted with water or an organic solvent to obtain an aqueous extract or organic extract of Antrodia camphorata using methods known to experts in this field of technology. Organic solvents include without limitation alcohols, such as methanol, ethanol or propanol, esters such as ethyl acetate, alkanes such as hexane, or alkyl halides, such as Harmattan, charatan. Among them, preferred is an alcohol, particularly preferred is ethanol. Aqueous or organic extracts from Antrodia camphorata was subjected to high performance liquid chromatography (HPLC) for separation and purification. Each fraction was collected and analyzed for anti-cancer effect. The active fractions from the anti-cancer effects analyzed the composition and subsequently investigated the activity against various tumor cells. The above approach has led to the identification of new compounds of the formula (1) and formula (2), inhibiting the growth of some tumor cells, was not detected previously in Antrodia camphorata, and which are not reported is camping in any previous publications. The compound 4-hydroxy-2,3-dimethoxy-6-methyl-5(3,7,11-trimethyl-dodeca-2,6,10-trienyl)-cyclohex-2 northward formula (2) is explained below as an example of the present invention. Antitumor effects of 4-hydroxy-2,3-dimethoxy-6-methyl-5(3,7,11-trimethyl-dodeca-2,b,10-trienyl)-cyclohex-2-Aenon was assessed by analysis with 3-(4, 5-dimethylthiazol-2-yl)-2, S-diphenyl of tetrazole bromide (MTT) test in accordance with the model for screening antitumor drugs of the National cancer Institute (NCI) at the level of survival of cells using cell lines such as breast cancer cells, liver cancer, prostate cancer and so like that. The above tests have proved that 4-hydroxy-2,3-dimethoxy-6-methyl-5(3,7,11-trimethyl-dodeca-2,6,10-trienyl)-cyclohex-2 northward reduces the levels of the survival cell lines breast cancer (MCF-7 and MDA-MB-231)cell lines hepatocellular carcinoma (ner 3V and ner G2) and cell lines of prostate cancer (LNCaP and DU-145), at the same time showing relatively low concentrations of proinvestirovany (IC50). The growth of breast cancer cells, liver cancer and prostate cancer was reduced 4-hydroxy-2,3-dimethoxy-6-methyl-5(3,7,11-trimethyl-dodeca-2,6,10-trienyl)-cyclohex-2-Aenon, which thus can be used to treat cancer, such as breast cancer, liver cancer, prostate cancer, and the like. The details of the examples describe what s next. Example 1 Selection 4-hydroxy-2,3-dimethoxy-6-methyl-5(3,7,11-trimethyl-dodeca-2,6,10-trienyl)-cyclohex-2-Aenon 100 g of mycelium, fruiting bodies, or a mixture thereof from Antrodia camphorata were placed in a flask. To the flask was added a suitable amount of water and alcohol (70-100% alcohol solution) and stirred at 20-25°C for at least 1 hour. The solution was filtered through a filter of 0.45 μm membrane, and collecting the filtrate in the quality of the extract. The filtrate from Antrodia camphorata were subjected to analysis by high performance liquid chromatography (HPLC). Separation was carried out on RP18 column; mobile phase consisted of methanol (A) and 0.1 to 0.5% acetic acid (V), gradient 0-10 min in 95%~20% B, 10-20 min in 20%~10%, 20-35 min in 10%~10% B, 35-40 min in 10%~95%, at a flow rate of 1 ml/min Flow arising from the column was monitored by a detector in the UV/visible region. Fractions collected at 25-30 min, were collected and concentrated to obtain 4-hydroxy-2,3-dimethoxy-6-methyl-5(3,7,11-trimethyl-dodeca-2,6,10-trienyl)-cyclohex-2-Aenon product as a pale yellow powder. Analysis of 4-hydroxy-2,3-dimethoxy-6-methyl-5(3,7,11-trimethyl-dodeca-2,6,10-trienyl)-cyclohex-2-Aenon showed the molecular formula C24H38O4, the molecular weight of 390, melting point 48°~52°C. the Study of the NMR spectrum showed that1N-NMK(CDC13)δ(ppm)=1.51, 1.67, 1.71, 1.75, 1.94, 2.03, 2.07, 2.22, 2.25, 3.68, 4.05, 5.07, and .14; 13C-NMK(CDC13)δ(ppm)=12.31, 16.1, 16.12, 17.67, 25.67, 26.44, 26.74, 27.00, 39.71, 39.81, 4.027, 43.34, 59.22, 60.59, 120.97, 123.84, 124.30, 131.32, 135.35, 135.92, 138.05, 160.45, and 197.12. Chemical structure of 4-hydroxy-2,3-dimethoxy-6-methyl-5(3,7,11-trimethyl-dodeca-2,6,10-trienyl)-cyclohex-2-may compared with a database of chemical compounds and was found similar patterns. These data confirmed that 4-hydroxy-2,3-dimethoxy-6-methyl-5(3,7,11-trimethyl-dodeca-2,6,10-trienyl)-cyclohex-2-the exact location is a new connection, which has not been previously reported. Example 2 Survival analysis in vitro effects against breast cancer Model screening of anticancer drugs NCI chose to test the anticancer effect of the compound of example 1 of the invention. Compound 4-hydroxy-2,3-dimethoxy-6-methyl-5(3,7,11-trimethyl-dodeca-2,6,10-trienyl)-cyclohex-2 northward from example 1 was added to the medium for cultivation of breast cancer cells human MCF-7 or MDA-MB-231, to analyze the survival of tumor cells. This analysis was carried out using the test with 3-(4, 5-dimethylthiazol-2-yl)-2, S-diphenyl of tetrazole bromide (MTT), usually used to measure cell proliferation, the percentage of viable cells and cytotoxicity. MTT is a yellow dye that is absorbed by living cells and restored to crimson and blue crystals formazan reductase with ccinet-tetrazole in mitochondria. Education formazan, therefore, used to evaluate and determine the level of survival of cells. Cancer cells human breast MCF-7 and MDA-MB-231 were cultured individually in media containing fetal calf serum for 24 hours. Proliferiruyuchei cells were washed once FBI, and then were treated with 1 × trypsin-EDTA and centrifuged at 1200 rpm for 5 minutes. The supernatant was discarded and the sediment cells resuspendable in 10 ml of fresh medium for cultivation when lightly shaken. The cells were placed in 96-well plate. Ethanol extracts of Antrodia camphorata (control group, the total extracts of Antrodia camphorata without purification) was added to each of 96 wells with the following concentrations: 30, 10, 3, 1, 0.3, 0.1 and 0.03 µg/ml, respectively, while 4-hydroxy-2,3-dimethoxy-6-methyl-5(3,7,11-trimethyl-dodeca-2,6,10-trienyl)-cyclohex-2-northward (experimental group) was added to each of 96 wells with the following concentrations: 30, 10, 3, 1, 0.3, 0.1 and 0.03 µg/ml, respectively. Cells were incubated at 37°C in 5% CO2incubator for 48 hours. MTT was added with a concentration of 2.5 mg/ml in each well in the dark and incubated for 4 hours, followed by addition of 100 ál of lytic buffer to stop the reaction. The tablets were analyzed on ELISA reader at a wavelength of 570 nm to determine the level of survival. C is achene concentrations of semi-inhibition (IC 50) was calculated and indicated in table 1.
From the results of table 1 shows that 4-hydroxy-2,3-dimethoxy-6-methyl-5(3,7,11-trimethyl-dodeca-2,6,10-trienyl)-cyclohex-2-the exact location is a potent inhibitor of the growth of cell lines of breast cancer person. The IC50 values of 4-hydroxy-2,3-dimethoxy-6-methyl-5(3,7,11-trimethyl-dodeca-2,6,10-trienyl)-cyclohex-2-Aenon for MCF-7 and MDA-MB-231 are 0,852 µg/ml and 1,031 µg/ml, respectively, significantly lower than the total extracts of Antrodia camphorata. Thus, 4-hydroxy-2,3-dimethoxy-6-methyl-5(3,7,11-Tr is methyl dodeca-2,6,10-trienyl)-cyclohex-2 northward from Antrodia camphorata can be used to inhibit the growth of breast cancer cells. Example 3 Additional in vitro study on adjuvant therapy of breast cancer cells The experiment was also performed in accordance with the in vitro model for screening anti-cancer drugs NCI. Cancer cells human breast MCF-7 and MDA-MB-231 were cultured individually in media containing fetal calf serum for 24 hours. Proliferiruyuchei cells were washed once FBI, and then were treated with 1 × trypsin-EDTA and centrifuged at 1200 rpm for 5 minutes. The supernatant was discarded and the sediment cells resuspendable in 10 ml of fresh medium for cultivation when lightly shaken. The cells were placed in 96-well plate after adding 0,0017 mg/ml Taxol, and processed within 72 hours. 4-hydroxy-2,3-dimethoxy-6-methyl-5(3,7,11 - trimethyl-dodeca-2,6,10-trienyl)-cyclohex-2 northward obtained in example 1 was added to each of 96 wells in the following concentrations: 0 mg/ml (control group); 30, 10, 3, 1, 0.3, 0.1 and 0.03 µg/ml (experimental group), respectively. Cells were incubated at 37°C in 5% CO2incubator for 48 hours. MTT was added with a concentration of 2.5 mg/ml in each well in the dark and incubated for 4 hours, followed by addition of 100 ál of lytic buffer to stop the reaction. The tablets were analyzed on ELISA reader at a wavelength of 570 nm to determine ur is una survival. Concentrations of semi-inhibition (IC50) was calculated and indicated in table 2.
From the results of table 2 shows that the values of the IC504-hydroxy-2,3-dimethoxy-6-methyl-5(3,7,11-trimethyl-dodeca-2,6,10-trienyl)-cyclohex-2-Aenon for MCF-7 and MDA-MB-231 declined to 0,009 mg/ml and to 0.011 μg/ml, respectively, after addition of Taxol. Thus, these results confirm the inhibitory activity of 4-hydroxy-2,3-dimethoxy-6-ethyl-5(3,7,11-trimethyl-dodeca-2,6,10-trienyl)-cyclohex-2-northward from Antrodia camphorate, which can be used to inhibit the growth of breast cancer cells, and show the best synergistic antitumor activity in combination with Taxol. Example 4 Survival analysis in vitro effects against liver cancer Model screening of anticancer drugs NCI also used for analysis of anticancer effects of compounds isolated in example 1 of the present invention. Compound 4-hydroxy-2,3-dimethoxy-6-methyl-5(3,7,11-trimethyl-dodeca-2,6,10-trienyl)-cyclohex-2 northward from example 1 was added to the medium for culturing cancer cells human liver, ner 3V or ner G2, to analyze the survival of tumor cells. Cancer cells human liver ner 3V and ner G2 were cultured individually in media containing fetal calf serum for 24 hours. Proliferiruyuchei cells were washed once FBI, and then were treated with 1 × trypsin-EDTA and centrifuged at 1200 rpm for 5 minutes. The supernatant was discarded and the sediment cells resuspendable in 10 ml of fresh medium for cultivation when lightly shaken. The cells were placed in 96-well plate. Ethanol extracts of Antrodia camphorata (control group, the total extracts of Antrodia camphorata without purification) was added to each of 96 wells with the following concentrations: 30, 10, 3, 1, 0.3, 0.1 and 0.03 µg/ml, respectively, while-hydroxy-2,3-dimethoxy-6-methyl-5(3,7,11-trimethyl-dodeca-2,6,10-trienyl)-cyclohex-2-northward (experimental group) was added to each of 96 wells with the following concentrations: 30, 10, 3, 1, 0.3, 0.1 and 0.03 µg/ml, respectively. Cells were incubated at 37°C in 5% CO2incubator for 48 hours. MTT was added with a concentration of 2.5 mg/ml in each well in the dark and incubated for 4 hours, followed by addition of 100 ál of lytic buffer to stop the reaction. The tablets were analyzed on ELISA reader at a wavelength of 570 nm to determine the level of survival. Concentrations of semi-inhibition (IC50) was calculated and indicated in table 3.
From the results of table 3 shows that 4-hydroxy-2,3-dimethoxy-6-methyl-5(3,7,11-trimethyl-dodeca-2,6,10-trienyl)-cyclohex-2-the exact location is a potent inhibitor of growth of cancer cell lines human liver. The values of the IC504-hydroxy-2,3-dimethoxy-6-methyl-5(3,7,11-trimethyl-dodeca-2,6,10-trienyl)-cyclohex-2-Aenon for ner 3V and ner G2 are 0.005 μg/ml and 1,679 µg/ml, respectively, significantly lower than the total extracts of Antrodia camphorata. Thus, 4-hydroxy-2,3-dimethoxy-6-methyl-5(3,7,11-trimethyl-dodeca-2,6,10-trienyl)-cyclohex-2 northward from Antrodia camphorata can be used to inhibit the growth of liver cancer cells. Example 5 Additional in vitro study on adjuvant therapy of liver cancer cells The experiment was also performed in accordance with the in vitro model for screening anti-cancer drugs NCI. Cancer cells human liver ner 3 and ner G2 were cultured separately in environments containing fetal calf serum for 24 hours. Proliferiruyuchei cells were washed once FBI, and then were treated with 1 × trypsin-EDTA and centrifuged at 1200 rpm for 5 minutes. The supernatant was discarded and the sediment cells resuspendable in 10 ml of fresh medium for cultivation when lightly shaken. Cells ner 3V processed 0,0043 mg/ml of Lovastatin for 72 hours, and ner G2 processed 0,0017 mg/ml Taxol for 72 hours, before being placed in a 96-well plate. 4-hydroxy-2,3-dimethoxy-6-methyl-5(3,7,11-trimethyl-dodeca-2,6,10-trienyl)-cyclohex-2 northward obtained in example 1 was added to each of 96 wells in the following concentrations: 0 mg/ml (control group); 30, 10, 3, 1, 0.3, 0.1 and 0.03 µg/ml (experimental group), respectively. Cells were incubated at 37°C in 5% CO2incubator for 48 hours. MTT was added with a concentration of 2.5 mg/ml in each well in the dark and incubated for 4 hours, followed by addition of 100 ál of lytic buffer to stop the reaction. The tablets were analyzed on ELISA reader at a wavelength of 570 nm to determine the level of survival. Concentrations of semi-inhibition (IC50) was calculated and indicated in table 4.
From the results of table 4 it is seen that the values of the IC504-hydroxy-2,3-dimethoxy-6-methyl-5(3,7,11-trimethyl-dodeca-2,6,10-trienyl)-cyclohex-2-Aenon for ner 3V and ner G2 decreased to 0.002 μg/ml and 0.008 μg/ml, respectively, after addition of the synergistic activity of Lovastatin and Taxol. Thus, these results confirm the inhibitory activity of 4-hydroxy-2,3-dimethoxy-6-methyl-5(3,7,11-trimethyl-dodeca-2,6,10-trienyl)-cyclohex-2-northward from Antrodia camphorata, which can be used to inhibit the growth of liver cancer cells, and showed better antitumor sinergetichesky the activity in combination with Taxol. Example 6 Survival analysis in vitro effects against prostate cancer Model screening of anticancer drugs NCI also used for analysis of anticancer effects of compounds isolated in example 1 of the present invention. Compound 4-hydroxy-2,3-dimethoxy-6-methyl-5(3,7,11-trimethyl-dodeca-2,6,10-trienyl)-peaklogix-2 northward from example 1 was added in the medium for cell culture prostate cancer human LNCaP or DU-145, to analyze the survival of tumor cells. Cancer cells of human prostate LNCaP or DU-145 were cultured individually in media containing fetal calf serum for 24 hours. Proliferiruyuchei cells were washed once FBI, and then were treated with 1 × trypsin-EDTA and centrifuged at 1200 rpm for 5 minutes. The supernatant was discarded and the sediment cells resuspendable in 10 ml of fresh medium for cultivation when lightly shaken. The cells were placed in 96-well plate. Ethanol extracts of Antrodia camphorata (control group, the total extracts of Antrodia camphorata without treatment) or 4-hydroxy-2,3-dimethoxy-6-methyl-5(3,7,11-trimethyl-dodeca-2,b,10-trienyl)-cyclohex-2-northward (experimental group) was added to each of 96 wells with the following concentrations: 30, 10, 3, 1, 0.3, 0.1 and 0.03 µg/ml, respectively. Cells were incubated at 37°C in 5% CO2incubator for 48 hours. MTT EXT is ulali with a concentration of 2.5 mg/ml in each well in the dark, and incubated for 4 hours, followed by addition of 100 ál of lytic buffer to stop the reaction. The tablets were analyzed on ELISA reader at a wavelength of 570 nm to determine the level of survival. Concentrations of semi-inhibition (IC50) was calculated and indicated in table 5.
From the results of table 5 it is seen that 4-hydroxy-2,3-dimethoxy-6-methyl-5(3,7,11-trimethyl-dodeca-2,6,10-trienyl)-cyclohex-2-the exact location is a potent inhibitor of the growth of cell lines of prostate cancer person. The values of the IC504-hydroxy-2,3-dimmock and-6-methyl-5(3,7,11-trimethyl-dodeca-2,6,10-trienyl)-cyclohex-2-Aenon for LNCaP and DU-145 are 2,378 µg/ml and 1,812 µg/ml respectively, significantly lower than the total extracts of Antrodia camphorata. Thus, 4-hydroxy-2,3-dimethoxy-6-methyl-5(3,7,11-trimethyl-dodeca-2,6,10-trienyl)-cyclohex-2 northward from Antrodia camphorata can be used to inhibit the growth of prostate cancer cells. Example 7 Additional in vitro study on adjuvant therapy of prostate cancer cells The experiment was also performed in accordance with the in vitro model for screening anti-cancer drugs NCI. Cancer cells of human prostate LNCaP and DU-145 were cultured individually in media containing fetal calf serum for 24 hours. Proliferiruyuchei cells were washed once FBI, and then were treated with 1 × trypsin-EDTA and centrifuged at 1200 rpm for 5 minutes. The supernatant was discarded and the sediment cells resuspendable in 10 ml of fresh medium for cultivation when lightly shaken. The LNCaP cells were treated 0,0017 mg/ml of Taxol within 72 hours, and DU-145 were treated 0,0043 mg/ml Taxol for 72 hours, before being placed in a 96-well plate. 4-hydroxy-2,3-dimethoxy-6-methyl-5(3,7,11-trimethyl-dodeca-2,6,10-trienyl)-cyclohex-2 northward obtained in example 1 was added to each of 96 wells in the following concentrations: 0 mg/ml (control group); 30, 10, 3, 1, 0.3, 0.1 and 0.03 µg/ml (experimental group), respectively. Cells were incubated at 37°C in 5% CO2Incubus is the Torah within 48 hours. MTT was added with a concentration of 2.5 mg/ml in each well in the dark and incubated for 4 hours, followed by addition of 100 ál of lytic buffer to stop the reaction. The tablets were analyzed on ELISA reader at a wavelength of 570 nm to determine the level of survival. Concentrations of semi-inhibition (IC50) was calculated and indicated in table 6.
From the results of table 6 shows that the values of the IC504-hydroxy-2,3-di is ethoxy-6-methyl-5(3,7,11-trimethyl-dodeca-2,6,10-trienyl)-cyclohex-2-Aenon for cancer cells of human prostate LNCaP and DU-145 decreased to 0,961 µg/ml and 0,5158 µg/ml accordingly, after combination with Taxol. Thus, these results confirm the inhibitory activity of 4-hydroxy-2,3-dimethoxy-6-methyl-5(3,7,11-trimethyl-dodeca-2,6,10-trienyl)-cyclohex-2-northward from Antrodia camphorate, which can be used to inhibit the growth of prostate cancer cells, and show the best synergistic antitumor activity in combination with Taxol. Example 8 Study of antioxidant activity in vitro Human low-density lipoprotein (LDL), oxidized copper ion (Cu2+), are widely used for evaluation of antioxidant activity of samples. The antioxidant activity of the sample is determined by the content of dienes in LDL after oxidation, and expressed in equivalents of Trolox using the standard curve, calculated on a water-soluble standard vitamin E to Trolox (a value of 1 antioxidant activity corresponds to 2 μm of Trolox). First preparing the following solutions: bidistilled water (negative control group), 5 mm sodium phosphate buffer (the NWF), 1 μm and 2 μm solution of Trolox (positive control group), and 40 µg/ml of 4-hydroxy-2,3-dimethoxy-6-methyl-5(3,7,11-trimethyl-dodeca-2,6,10-trienyl)-cyclohex-2-Aenon Veselinovo in example 1. The concentration of LDL cholesterol (LDL-C) was determined using the method of the enzymatic reaction, ova diluted to 0.1-0.25 mg/ml with 5 mm the NWF. One hundredth μl of LDL was added into each well of 96-well quartz tablet, followed by the addition of the above-mentioned Trolox and connections allocated in example 1. To induce oxidation, was added standardized oxidizing agent CuSO4to a final concentration of 5 μm in each 250 ál of the hole. The tablet was analyzed on ELISA reader at a wavelength of 232 nm at 37°C for 12 hours. The sampling time was 15 minutes. The results are shown in table 7.
Note 1: the Time lag phase (Tlag, min) was determined as the intersection of the lag-phase with the phase distribution of the absorption at 234 nm. ΔTlag(min) was defined as the time difference between Tlag and Tlag0for each sample. Note 2: it is Considered that the connection has antioxidant ability, when the value of antioxidant activity greater than 0.5. From the results of table 7 shows that the value of antioxidant activity of 4-hydroxy-2,3-dimethoxy-6-methyl-5(3,7,11-trimethyl-dodeca-2,6,10-trienyl)-cyclohex-2-Aenon is 1.3, which is much larger than the standard value of 0.5. Thus, the compounds of the invention possess antioxidant activity that can be used for healthy food, diets and drinks, the effect is nski products and cosmetics, and to have significant beneficial effects on human health due to their ability to prevention of cardiovascular disease or mutation of cells. 1. The compound having formula (1): 2. The compound according to claim 1 in which the compound isolated from Antrodia camphorata. 3. The compound according to claim 2, in which the connection is isolated from the extracts of Antrodia camphorata, obtained using organic solvents. 4. The compound according to claim 3, in which organic solvents selected from the group consisting of alcohols, esters, alkanes and alkyl halides. 5. The compound according to claim 4, in which the alcohol is ethanol. 6. The compound according to claim 2, in which the compound isolated from aqueous extracts of Antrodia camphorata. 7. The compound according to claim 1, which represents a 4-hydroxy-2,3-dimethoxy-6-methyl-5(3,7,11-trimethyl-dodeca-2,6,10-trienyl)-cyclohex-2 northward. 8. The compound according to claim 1 or 7, exhibiting antioxidant activity. 9. The method of suppressing the growth of breast cancer cells, comprising applying a compound according to claim 1 or 7. 10. The method according to claim 9, in which the compound isolated from Antrodia camphorata. 11. The method according to claim 10, in which a compound selected from EXT the acts of Antrodia camphorata, obtained using organic solvents. 12. The method according to claim 11, in which the organic solvent is selected from the group consisting of alcohols, esters, alkanes and alkyl halides. 13. The method according to item 12, in which the organic solvent is ethanol. 14. The method according to claim 10, in which the compound isolated from aqueous extracts of Antrodia camphorata. 15. The method according to claim 9, wherein the breast cancer cells related to cell line MCF-7 or MDA-MB-231. 16. A method of inhibiting the growth of liver cancer cells, comprising applying a compound according to claim 1 or 7. 17. The method according to clause 16, in which the compound isolated from Antrodia camphorata. 18. The method according to 17, in which the compound selected from extracts of Antrodia camphorata, obtained using organic solvents. 19. The method according to p, in which organic solvents selected from the group consisting of alcohols, esters, alkanes and alkyl halides. 20. The method according to claim 19, in which the organic solvent is ethanol. 21. The method according to 17, in which the connection is isolated from the aqueous extracts of Antrodia camphorata. 22. The method according to clause 16, in which the cells of liver cancer related to cell lines ner 3 or ner G2. 23. A method of inhibiting the growth of prostate cancer cells, comprising applying a compound according to claim 1 or 7. 24. The method according to item 23, in which the compound isolated from Antrodia camphorata. 25. The method according to the .24, in which compound selected from extracts of Antrodia camphorata, obtained using organic solvents. 26. The method according A.25, in which organic solvents selected from the group consisting of alcohols, esters, alkanes and alkyl halides. 27. The method according to p, in which the organic solvent is ethanol. 28. The method according to paragraph 24, in which the connection is isolated from the aqueous extracts of Antrodia camphorata. 29. The method according to item 23, wherein the prostate cancer cells include cell lines LNCaP or DU145. 30. Pharmaceutical composition for inhibiting the growth of tumor cells comprising an active dose of a compound according to claim 1 and a pharmaceutically suitable carrier, in which tumor cells are selected from the group consisting of breast cancer, liver cancer and prostate cancer. 31. Pharmaceutical composition for inhibiting the growth of tumor cells comprising an active dose of a compound according to claim 7 and a pharmaceutically suitable carrier, in which tumor cells are selected from the group consisting of breast cancer, liver cancer and prostate cancer.
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