CN100410226C - Quinone compounds, and their preparing method and antitumour use - Google Patents
Quinone compounds, and their preparing method and antitumour use Download PDFInfo
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- CN100410226C CN100410226C CNB200610035637XA CN200610035637A CN100410226C CN 100410226 C CN100410226 C CN 100410226C CN B200610035637X A CNB200610035637X A CN B200610035637XA CN 200610035637 A CN200610035637 A CN 200610035637A CN 100410226 C CN100410226 C CN 100410226C
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- 150000004053 quinones Chemical class 0.000 title claims abstract description 26
- 230000000259 anti-tumor effect Effects 0.000 title abstract description 8
- 238000000034 method Methods 0.000 title description 3
- 238000002360 preparation method Methods 0.000 claims abstract description 13
- 241001249943 Halorosellinia sp. Species 0.000 claims abstract description 8
- 238000000855 fermentation Methods 0.000 claims abstract description 8
- 230000004151 fermentation Effects 0.000 claims abstract description 8
- 238000000605 extraction Methods 0.000 claims abstract description 6
- 239000002246 antineoplastic agent Substances 0.000 claims abstract description 3
- 229940041181 antineoplastic drug Drugs 0.000 claims abstract description 3
- XEKOWRVHYACXOJ-UHFFFAOYSA-N Ethyl acetate Chemical compound CCOC(C)=O XEKOWRVHYACXOJ-UHFFFAOYSA-N 0.000 claims description 30
- RTZKZFJDLAIYFH-UHFFFAOYSA-N Diethyl ether Chemical compound CCOCC RTZKZFJDLAIYFH-UHFFFAOYSA-N 0.000 claims description 20
- WFDIJRYMOXRFFG-UHFFFAOYSA-N Acetic anhydride Chemical compound CC(=O)OC(C)=O WFDIJRYMOXRFFG-UHFFFAOYSA-N 0.000 claims description 18
- 239000003208 petroleum Substances 0.000 claims description 16
- 235000015097 nutrients Nutrition 0.000 claims description 14
- JUJWROOIHBZHMG-UHFFFAOYSA-N Pyridine Chemical compound C1=CC=NC=C1 JUJWROOIHBZHMG-UHFFFAOYSA-N 0.000 claims description 12
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 9
- 238000001953 recrystallisation Methods 0.000 claims description 7
- HEDRZPFGACZZDS-UHFFFAOYSA-N Chloroform Chemical compound ClC(Cl)Cl HEDRZPFGACZZDS-UHFFFAOYSA-N 0.000 claims description 6
- WQZGKKKJIJFFOK-GASJEMHNSA-N Glucose Natural products OC[C@H]1OC(O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-GASJEMHNSA-N 0.000 claims description 6
- 239000001888 Peptone Substances 0.000 claims description 6
- 108010080698 Peptones Proteins 0.000 claims description 6
- DPDMMXDBJGCCQC-UHFFFAOYSA-N [Na].[Cl] Chemical compound [Na].[Cl] DPDMMXDBJGCCQC-UHFFFAOYSA-N 0.000 claims description 6
- 230000001580 bacterial effect Effects 0.000 claims description 6
- 229940041514 candida albicans extract Drugs 0.000 claims description 6
- 238000004440 column chromatography Methods 0.000 claims description 6
- 239000003480 eluent Substances 0.000 claims description 6
- 238000010828 elution Methods 0.000 claims description 6
- 239000008103 glucose Substances 0.000 claims description 6
- 235000019319 peptone Nutrition 0.000 claims description 6
- UMJSCPRVCHMLSP-UHFFFAOYSA-N pyridine Natural products COC1=CC=CN=C1 UMJSCPRVCHMLSP-UHFFFAOYSA-N 0.000 claims description 6
- 238000012360 testing method Methods 0.000 claims description 6
- 239000012138 yeast extract Substances 0.000 claims description 6
- 239000013078 crystal Substances 0.000 claims description 5
- 238000001914 filtration Methods 0.000 claims description 5
- FRXSZNDVFUDTIR-UHFFFAOYSA-N 6-methoxy-1,2,3,4-tetrahydroquinoline Chemical compound N1CCCC2=CC(OC)=CC=C21 FRXSZNDVFUDTIR-UHFFFAOYSA-N 0.000 claims description 3
- 229920001817 Agar Polymers 0.000 claims description 3
- 239000004952 Polyamide Substances 0.000 claims description 3
- VYPSYNLAJGMNEJ-UHFFFAOYSA-N Silicium dioxide Chemical compound O=[Si]=O VYPSYNLAJGMNEJ-UHFFFAOYSA-N 0.000 claims description 3
- UGAPHEBNTGUMBB-UHFFFAOYSA-N acetic acid;ethyl acetate Chemical compound CC(O)=O.CCOC(C)=O UGAPHEBNTGUMBB-UHFFFAOYSA-N 0.000 claims description 3
- 239000008272 agar Substances 0.000 claims description 3
- 238000013375 chromatographic separation Methods 0.000 claims description 3
- 239000012141 concentrate Substances 0.000 claims description 3
- 230000000694 effects Effects 0.000 claims description 3
- SRCZQMGIVIYBBJ-UHFFFAOYSA-N ethoxyethane;ethyl acetate Chemical compound CCOCC.CCOC(C)=O SRCZQMGIVIYBBJ-UHFFFAOYSA-N 0.000 claims description 3
- 239000002024 ethyl acetate extract Substances 0.000 claims description 3
- 238000003810 ethyl acetate extraction Methods 0.000 claims description 3
- 239000000284 extract Substances 0.000 claims description 3
- 239000008187 granular material Substances 0.000 claims description 3
- 229920002647 polyamide Polymers 0.000 claims description 3
- 238000011218 seed culture Methods 0.000 claims description 3
- 239000000741 silica gel Substances 0.000 claims description 3
- 229910002027 silica gel Inorganic materials 0.000 claims description 3
- 238000010898 silica gel chromatography Methods 0.000 claims description 3
- 229960001866 silicon dioxide Drugs 0.000 claims description 3
- 238000003756 stirring Methods 0.000 claims description 3
- 150000001875 compounds Chemical class 0.000 abstract description 7
- 238000000926 separation method Methods 0.000 abstract description 3
- 210000004881 tumor cell Anatomy 0.000 abstract description 3
- 230000004614 tumor growth Effects 0.000 abstract description 2
- 230000021736 acetylation Effects 0.000 abstract 1
- 238000006640 acetylation reaction Methods 0.000 abstract 1
- 210000004027 cell Anatomy 0.000 description 32
- 239000000243 solution Substances 0.000 description 11
- 241000233866 Fungi Species 0.000 description 10
- 239000003814 drug Substances 0.000 description 9
- 206010006187 Breast cancer Diseases 0.000 description 6
- 208000026310 Breast neoplasm Diseases 0.000 description 6
- 206010073071 hepatocellular carcinoma Diseases 0.000 description 6
- 240000002044 Rhizophora apiculata Species 0.000 description 5
- 244000005700 microbiome Species 0.000 description 5
- 241000196324 Embryophyta Species 0.000 description 4
- 229940079593 drug Drugs 0.000 description 4
- IAZDPXIOMUYVGZ-UHFFFAOYSA-N Dimethylsulphoxide Chemical compound CS(C)=O IAZDPXIOMUYVGZ-UHFFFAOYSA-N 0.000 description 3
- 206010058467 Lung neoplasm malignant Diseases 0.000 description 3
- 206010028980 Neoplasm Diseases 0.000 description 3
- 125000002777 acetyl group Chemical group [H]C([H])([H])C(*)=O 0.000 description 3
- 201000011510 cancer Diseases 0.000 description 3
- 230000002401 inhibitory effect Effects 0.000 description 3
- 201000005202 lung cancer Diseases 0.000 description 3
- 208000020816 lung neoplasm Diseases 0.000 description 3
- 239000002207 metabolite Substances 0.000 description 3
- 210000000214 mouth Anatomy 0.000 description 3
- 238000012549 training Methods 0.000 description 3
- 238000001644 13C nuclear magnetic resonance spectroscopy Methods 0.000 description 2
- 238000005160 1H NMR spectroscopy Methods 0.000 description 2
- VHYFNPMBLIVWCW-UHFFFAOYSA-N 4-Dimethylaminopyridine Chemical compound CN(C)C1=CC=NC=C1 VHYFNPMBLIVWCW-UHFFFAOYSA-N 0.000 description 2
- IJGRMHOSHXDMSA-UHFFFAOYSA-N Atomic nitrogen Chemical compound N#N IJGRMHOSHXDMSA-UHFFFAOYSA-N 0.000 description 2
- WORJEOGGNQDSOE-UHFFFAOYSA-N chloroform;methanol Chemical compound OC.ClC(Cl)Cl WORJEOGGNQDSOE-UHFFFAOYSA-N 0.000 description 2
- 239000000463 material Substances 0.000 description 2
- 229930014626 natural product Natural products 0.000 description 2
- 239000002953 phosphate buffered saline Substances 0.000 description 2
- 238000004321 preservation Methods 0.000 description 2
- 239000000047 product Substances 0.000 description 2
- 150000003839 salts Chemical class 0.000 description 2
- 230000001954 sterilising effect Effects 0.000 description 2
- 238000004659 sterilization and disinfection Methods 0.000 description 2
- 238000010998 test method Methods 0.000 description 2
- 239000003643 water by type Substances 0.000 description 2
- ZCWPHDXKEDBCER-UHFFFAOYSA-N 2,5-diphenyl-2h-tetrazol-2-ium;bromide Chemical compound [Br-].C1=CC=CC=C1C1=[NH+]N(C=2C=CC=CC=2)N=N1 ZCWPHDXKEDBCER-UHFFFAOYSA-N 0.000 description 1
- 229960000549 4-dimethylaminophenol Drugs 0.000 description 1
- 241000894006 Bacteria Species 0.000 description 1
- 208000024172 Cardiovascular disease Diseases 0.000 description 1
- 241000195493 Cryptophyta Species 0.000 description 1
- 108090000790 Enzymes Proteins 0.000 description 1
- 102000004190 Enzymes Human genes 0.000 description 1
- 240000003793 Rhizophora mangle Species 0.000 description 1
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 description 1
- -1 acetyl quinones Chemical class 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 238000009395 breeding Methods 0.000 description 1
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- 230000007812 deficiency Effects 0.000 description 1
- 238000001514 detection method Methods 0.000 description 1
- LOKCTEFSRHRXRJ-UHFFFAOYSA-I dipotassium trisodium dihydrogen phosphate hydrogen phosphate dichloride Chemical compound P(=O)(O)(O)[O-].[K+].P(=O)(O)([O-])[O-].[Na+].[Na+].[Cl-].[K+].[Cl-].[Na+] LOKCTEFSRHRXRJ-UHFFFAOYSA-I 0.000 description 1
- 238000002451 electron ionisation mass spectrometry Methods 0.000 description 1
- 238000004945 emulsification Methods 0.000 description 1
- 238000005516 engineering process Methods 0.000 description 1
- 238000004992 fast atom bombardment mass spectroscopy Methods 0.000 description 1
- 230000012010 growth Effects 0.000 description 1
- 238000010438 heat treatment Methods 0.000 description 1
- 230000001900 immune effect Effects 0.000 description 1
- 239000002955 immunomodulating agent Substances 0.000 description 1
- 230000002584 immunomodulator Effects 0.000 description 1
- 229940121354 immunomodulator Drugs 0.000 description 1
- 238000000338 in vitro Methods 0.000 description 1
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Landscapes
- Preparation Of Compounds By Using Micro-Organisms (AREA)
- Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
Abstract
The present invention relates to the field of compounds in quinone class, particularly to compounds in the quinone class having an anti-tumor activity and a preparation method and an application thereof. In the present invention, the red compound in the quinone class and an acetylation product thereof obtained by the extraction separation from a fermentation culture object of a marine fungus Halorosellinia sp. 1403 in South China Sea can effectively inhibit the growth of tumor cell strains. The present invention can be used for developing anti-tumor drugs and have a wide application prospect.
Description
Technical field
The present invention relates to the quinones field, be specifically related to quinones and preparation method thereof and application that a class has anti-tumor activity.
Background technology
Marine microorganism produces the active metabolite of novel structure with its unique pathways metabolism, and its meta-bolites becomes the abundant source of medicine, the meta-bolites of marine microorganism is pharmaceutical use widely also, as antitumor, treatment cardiovascular disorder, immunomodulator etc.Because the singularity of ocean environment, marine microorganism is of a great variety, wide material sources, screening pick-up rate height, according to the report of John professor, found that marine microorganism was one of main source in the marine natural product new compound in 2003 at 2005 " Natural Product Reports ".
Plant endogenesis epiphyte (Endophytic fungi) is lived in the tissue of higher plant, it is the natural moiety in the plant microecology system, set up harmonious symbiotic relationship with the host during evolution, endogenetic fungus is of a great variety at a conservative estimate, and nearly 1.5 * 10
6Kind because quantity is huge, and and other biological between ecological relationship closely, make this class fungi become potential and have the source that produces abundant secondary metabolite.
Mangrove forest is the plant that is grown in the torrid zone and tropical coastal tideland slob, and mangrove forest ecological system is one of the richest diversity, marine ecosystem that productivity is the highest in the world.Outer saline waters nutritive deficiency, but the mangrove swamp district is typical salty slightly waters band, compile the various inorganic salt and the organism that bring from upstream and ocean and add dropping of the dry branches and fallen leaves of mangrove plant body own, competent vegetable material is arranged, abundant planktonic algae and planktonic organism are arranged, and these conditions just are being well-suited for the breeding of microorganism (bacterium and fungi).In more than ten years in the past, the mangrove forest habitat is proved to be the abundant source of the new kind of fungi, and this has formed second largest ecological subclass of thalassiomycetes.
Summary of the invention
The object of the present invention is to provide a class to derive from the quinones with anti-tumor activity and the acetyl derivatives thereof of ocean mangrove fungi.
Another object of the present invention provides the preparation method of above-mentioned quinones.
Further purpose of the present invention provides the application of above-mentioned quinones in the preparation antitumor drug.
The present invention's extraction separation from the fermenting culture of South Sea thalassiomycetes Halorosellinia sp.1403 (hereinafter to be referred as fungi 1403) obtains a kind of quinones (1403C) of redness, and its structural formula is suc as formula (I): R wherein
1, R
2, R
3, R
4, R
5All be H.
Quinones 1403C under the effect of catalyzer, obtains a series of ethanoyl through aceticanhydride/pyridine acetylize and replaces the acetyl quinones that replaces to penta-acetyl, and its structural formula is suc as formula (I), wherein R
1, R
2, R
3, R
4, R
5Be H or CH
3CO, R
1, R
2, R
3, R
4, R
5Be not H simultaneously.
The used fungi 1403 of the present invention has been preserved in Chinese typical culture collection center (CCTCC, Chinese Wuhan University are in the school), and preserving number is CCTCC NO:M201018, and preservation day is April 23 calendar year 2001.
Quinones 1403C of the present invention can obtain by extraction separation from the fermentation culture of fungi 1403, and preparation method's concrete steps are as follows:
(1) seed culture of fungi Halorosellinia sp.1403CCTCC NO:M 201018: substratum is by weight: glucose 0.5-1.5, yeast extract 0.05-0.15, peptone 0.1-0.3, agar 1-1.5, sodium-chlor 3-5, water 100 is made the test tube slant, the picking bacterial strain inserts the inclined-plane, cultivates 5-7 days for 30-35 ℃;
(2) fermentation culture of fungi Halorosellinia sp.1403CCTCC NO:M 201018: fermention medium is by weight: glucose 5-15, yeast extract 1-4, peptone 0.5-4, sodium-chlor 3-5, water 100, cultured bacterial strain in the inclined-plane is chosen into fermention medium, left standstill 1-2 month in room temperature 25-35 ℃;
(3) above-mentioned cultured filtering fermentation liquor is removed thalline and obtain nutrient solution;
(4) heating of fungi 1403 nutrient solutions concentrates (temperature is no more than 50 ℃) 1/20-1/5 to the stoste volume, with ethyl acetate extraction repeatedly, concentrating acetic acid ethyl acetate extract, carry out chromatographic separation in silicagel column, is the eluent gradient elution with petroleum ether-ethyl acetate-methyl alcohol;
(5) nutrient solution medicinal extract is collected the ethyl acetate/petroleum ether elutriant of 50%-100% through behind the column chromatography, and 70% ethyl acetate/petroleum ether is an elutriant, separates with polyamide column chromatography again, and recrystallization purifying promptly obtains red granules shape crystal 1403C.
The acetylize quinones is to obtain acetylate by 1403C through aceticanhydride/pyridine acetylize, and preparation method's concrete steps are as follows:
Get dried 1403C, under the effect of catalyzer, add exsiccant pyridine, add acetic anhydride again, under 10~80 ℃, stir 30min~15h, add water, chloroform extraction; Through silica gel column chromatography, petroleum ether-ethyl acetate is the eluent gradient elution, collects 5%-80% ethyl acetate/petroleum ether elutriant, recrystallization purifying, and a series of ethanoyl that obtain 1403C replace the acetylize quinones that replaces to penta-acetyl.
20~50% ethyl acetate/petroleum ether elutriants, the chloroform-methanol recrystallization obtains the product 1403C-3Ac (R of orange-yellow crystal triacetylization
1, R
2, R
4CH
3CO, R
3, R
5Be H).
Compared with prior art, the present invention has following beneficial effect: the present invention is through evidence, and quinones 1403-C and acetylate thereof all can effectively suppress the growth of tumor cell line, can be used for developing anti-tumor medicaments, have a extensive future.
Embodiment
The preparation of embodiment 1 compound 1403-C and acetylate thereof
(1) seed culture of fungi Halorosellinia sp.1403:
Substratum is by weight: glucose 0.5-1.5, and yeast extract 0.05-0.15, peptone 0.1-0.3, agar 1-1.5, sodium-chlor 3-5, water 100 is made the test tube slant, and the picking bacterial strain inserts the inclined-plane, cultivates 5-7 days for 30-35 ℃;
(2) fermentation culture of fungi Halorosellinia sp.1403:
Fermention medium is by weight: glucose 5-15, and yeast extract 1-4, peptone 0.5-4, sodium-chlor 3-5, water 100 chooses cultured bacterial strain in the inclined-plane into fermention medium, leaves standstill 1-2 month in room temperature 25-35 ℃;
(3) above-mentioned cultured filtering fermentation liquor is removed thalline and obtain nutrient solution;
(4) be no more than under 70 ℃ of conditions in temperature, nutrient solution is heated the 1/20-1/5 that is concentrated into the stoste volume, with ethyl acetate extraction repeatedly, concentrate acetic acid ethyl acetate extract, carrying out chromatographic separation in silicagel column, is the eluent gradient elution with petroleum ether-ethyl acetate-methyl alcohol.
(5) nutrient solution medicinal extract is collected the ethyl acetate/petroleum ether elutriant of 50%-100% through behind the column chromatography, and 70% ethyl acetate/petroleum ether is an elutriant, separates with polyamide column chromatography again, and recrystallization purifying promptly obtains red granules shape crystal 1403C.
Acetylate 1403C-3Ac is by 1403C, and at catalyzer, through the triacetyl product that aceticanhydride/pyridine acetylize obtains, preparation method's concrete steps are as follows:
Get exsiccant 1403C, add exsiccant pyridine and acetic anhydride, a spot of DMAP under the room temperature, stirs 3-6h, adds water, chloroform extraction.Through silica gel column chromatography, petroleum ether-ethyl acetate is the eluent gradient elution, collects 20~50% ethyl acetate/petroleum ether elutriants, and the chloroform-methanol recrystallization obtains orange-yellow crystal 1403-C-Ac-3.
The testing data of 1403C:
FABMS:337(M+1),m/z:55.EA(w/%,):C 56.56,H 4.617。Calculate straight (C
16H
16O
8): C 57.14, and H 4.76,232 ℃ of .IR v/cm of mp
-1(KBr): 3514,3479,3374,3029,2987,2938,2896,1595,1475,1440,1398,1370,1300,1200,1208,1138,1082,997,948,864,821,632,554,491.
1HNMR (500Mz, CDCl
3, TMS): 13.35 (s, OH-9), 12.62 (s, OH-10), 6.46 (s, H-3), 4.77 (t, 4.5,4.5Hz, H-7, OH), 3.92 (s, CH
3-16), 3.54 (t, 4.5,4.5Hz, H-5), 2.74 (d, 18Hz, H-8a), 2.67 (d, 18Hz, H-8b), 1.24 (s, CH
3-15).
13CNMR (CDCl
3): 183.3 (C-4), 176.5 (C-1), 160.9 (C-10), 160.9 (C-2), 160.2 (C-9), 139.5 (C-11), 136.8 (C-12), 109.8 (C-14), 109.6 (C-3), 107.3 (C-13), 76.3 (C-7), 69.2 (C-6), 68.2 (C-5), 56.8 (C-16), 34.8 (C-8), 25.5 (C-15).
The testing data of 1403C-3Ac:
EI-MS:462,mp 259~260℃
1HNMR(500Mz,CDCl
3,TMS):6.026(H-3),5.287(dd,),3.98(d),3.07(dd,15,15HZ),2.9(dd,15,15HZ),1.361(H-15),2.12(CH3-22),2.37(CH
3-21),2.12(CH
3-20),12.65(OH-9),6.19(OH-7);
13CNMR(CDCl
3):184.8(C-4),182.7(C-1),170.5(C-18),170.4(C-19),169.9(C-17),158.9(C-9),158.9(C-2),132.4(C-11),132.1(C-12),113.0(C-13),112.0(C-14),111.6(C-3),73.6(C-6),70.5(C-7),69.4(C-8),35.2(C-5),29.7(C-15),21.1(C-22),20.8(C-21),20.8(C-20).
Embodiment 2 MTT reduction method detection compound 1403C and 1403C-3Ac anti-tumor activities
Test
1, material:
1.1 four Cuo salt (MTT): (3-(4 for phosphate buffered saline buffer (PBS) the dissolving MTT of usefulness 0.01mol/L, 5-dimethythiazol-z-yl) 2,5-diphenyltetrazolium bromide, SIGMA) final concentration 5mg/mL, filtration sterilization, 4 ℃ keep in Dark Place after the packing.
1.2 the preparation of target cell (is example with the HepG2 cell): with the recovery and the cultivation of HepG2 cell:
A. the frozen pipe of taking-up human hepatoma cell strain HepG2 cell from liquid nitrogen container is inserted rapidly in 37 ℃ of water-baths, does not stop to shake to make it to dissolve rapidly, and aseptic technique moves in the centrifuge tube;
B. add full nutrient solution to 10mL, the centrifugal 5s of 1000rmp abandons supernatant;
C. repeat above operation once;
D. move in the culturing bottle 5%CO after making the cell mixing with full nutrient solution piping and druming
2, 37 ℃ of cultivations;
E. the observation of cell growing state is in time changed nutrient solution, divides bottle.
1.3 cell counting:
A. choose the logarithmic phase cell, trysinization, the full cultivation stops, and moves in the centrifuge tube, adds full the training to 10mL;
B. get and one after another drop ofly go in the tally one side groove, the total cellular score of microscopically counting four big lattice, divided by 4 takes advantage of 10
4, be every milliliter of contained cell count of nutrient solution;
C. adjust cell count to 1 * 10
5/ mL;
1.4 the preparation of compd E and F: get compd E and F respectively and join in the full training, adjusting concentration is 500 μ g/mL, ultrasonic emulsification, filtration sterilization, 4 ℃ of preservations.
2. test method (is example with the HepG2 cell)
A.96 each hole of orifice plate adds HepG2 cell 100 μ L (1 * 10
5/ mL), 5%CO
2, cultivate 4hr for 37 ℃.
B. add different concns study subject 100 μ L, contrast adds full training 100 μ L, continues to cultivate 48hr.
C. add each 10 μ L of MTT (5mg/mL), continue to cultivate 4hr.
D. remove nutrient solution.Every hole adds DMSO 100 μ L, and the 5-10min that vibrates gently makes the particle dissolving.
E. enzyme linked immunological instrument 570nm measures every hole OD value down.
F. calculate inhibiting rate
The average OD value that the average OD value that tumor cell destruction %=[(control group is measured-dosing group is measured)/the average OD value of control group mensuration] * 100%
G. with the logarithm mapping of inhibiting rate, try to achieve IC to drug level
50Value
With lg c is X-coordinate, and inhibiting rate is an ordinate zou, tries to achieve IC
50Value
Lung cancer cell line A549, hepatoma cell strain Hep-2, cancer cell of oral cavity strain KB, breast cancer cell strain MCF-7, the test method of the anti-medicine breast cancer cell of multiple medicines strain MCF-7/Adr is the same.
3. test-results
Test-results shows that compound 1403C and 1403C-3Ac all can effectively suppress lung cancer cell line A549, hepatoma cell strain Hep-2 and hepatoma cell strain Hep G2, cancer cell of oral cavity strain KB, breast cancer cell strain MCF-7, the growth of the anti-medicine breast cancer cell of multiple medicines strain MCF-7/Adr.1403C and 1403C-3Ac are to the IC of these JEG-3
50Value (μ g/ml) sees Table 1.
The cell in vitro poison test-results (IC of table 1,1403-C and 1403-C-Ac-3
50μ g/mL)
Cell strain | 1403C | 1403C-3Ac |
Lung cancer cell line A549 | 2.44 | 0.69 |
Hepatoma cell strain Hep-2 | 3.15 | 1.00 |
Hepatoma cell strain Hep G2 | 4.41 | 0.85 |
Cancer cell of oral cavity strain KB | 3.15 | 0.92 |
Breast cancer cell strain MCF-7 | 4.76 | 3.02 |
The anti-medicine breast cancer cell of multiple medicines strain MCF-7/Adr | 5.45 | 2.97 |
Claims (3)
2. the preparation method of the described quinones of claim 1 is characterized in that may further comprise the steps:
(1) seed culture of fungi Halorosellinia sp.1403: substratum is by weight: glucose 0.5-1.5, yeast extract 0.05-0.15, peptone 0.1-0.3, agar 1-1.5, sodium-chlor 3-5, water 100; Make the test tube slant, the picking bacterial strain inserts the inclined-plane, cultivates 5-7 days for 30-35 ℃;
(2) fermentation culture of fungi Halorosellinia sp.1403: fermention medium is by weight: glucose 5-15, yeast extract 1-4, peptone 0.5-4, sodium-chlor 3-5, water 100; Cultured bacterial strain in the inclined-plane is chosen into fermention medium, left standstill 1-2 month in room temperature 25-35 ℃;
(3) above-mentioned cultured filtering fermentation liquor is removed thalline and obtain nutrient solution;
(4) be no more than under 50 ℃ of conditions in temperature, nutrient solution is heated the 1/20-1/5 that is concentrated into the stoste volume, with ethyl acetate extraction repeatedly, concentrate acetic acid ethyl acetate extract, carrying out chromatographic separation in silicagel column, is the eluent gradient elution with petroleum ether-ethyl acetate-methyl alcohol;
(5) nutrient solution medicinal extract is collected the ethyl acetate/petroleum ether elutriant of 50%-100% through behind the column chromatography, and 70% ethyl acetate/petroleum ether is an elutriant, separates with polyamide column chromatography again, and recrystallization purifying promptly obtains red granules shape crystal 1403C; 1403C is meant that structural formula is suc as formula (I), R
1, R
2, R
3, R
4, R
5It all is the quinones of H;
(6) get dried 1403C, under the effect of catalyzer, add exsiccant pyridine, add acetic anhydride again, under 10~80 ℃, stir 30min~12h, add water, chloroform extraction; Through silica gel column chromatography, petroleum ether-ethyl acetate is the eluent gradient elution, collects 5%-80% ethyl acetate/petroleum ether elutriant, and recrystallization purifying obtains the acetylate of 1403C.
3. the application of the described quinones of claim 1 in the preparation antitumor drug.
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