CN107298671A - Come from the secalonic acid H of penicillium oxalicum and prepare the application of anti-human colon cancer drug - Google Patents
Come from the secalonic acid H of penicillium oxalicum and prepare the application of anti-human colon cancer drug Download PDFInfo
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- CN107298671A CN107298671A CN201710460067.7A CN201710460067A CN107298671A CN 107298671 A CN107298671 A CN 107298671A CN 201710460067 A CN201710460067 A CN 201710460067A CN 107298671 A CN107298671 A CN 107298671A
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- human colon
- penicillium oxalicum
- colon cancer
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- 241000985513 Penicillium oxalicum Species 0.000 title claims abstract description 23
- 206010009944 Colon cancer Diseases 0.000 title claims abstract description 16
- 208000029742 colonic neoplasm Diseases 0.000 title claims abstract description 15
- 239000003560 cancer drug Substances 0.000 title claims abstract description 7
- ZPWLYPAANXFZTB-NRRMYFFQSA-N methyl (3S,4R,4aR)-7-[3-(3,6-dihydroxy-2-methoxycarbonyl-4-methylbenzoyl)-2,4-dihydroxyphenyl]-4,8,9-trihydroxy-3-methyl-1-oxo-3,4-dihydro-2H-xanthene-4a-carboxylate Chemical compound COC(=O)c1c(O)c(C)cc(O)c1C(=O)c1c(O)ccc(c1O)-c1ccc2O[C@]3([C@H](O)[C@@H](C)CC(=O)C3=C(O)c2c1O)C(=O)OC ZPWLYPAANXFZTB-NRRMYFFQSA-N 0.000 title abstract description 6
- 150000001875 compounds Chemical class 0.000 claims abstract description 28
- 239000003814 drug Substances 0.000 claims abstract description 9
- 230000035755 proliferation Effects 0.000 claims abstract description 8
- 210000001072 colon Anatomy 0.000 claims abstract description 7
- 238000002360 preparation method Methods 0.000 claims description 3
- 230000000694 effects Effects 0.000 abstract description 9
- 238000011160 research Methods 0.000 abstract description 5
- 230000000259 anti-tumor effect Effects 0.000 abstract description 3
- 230000000118 anti-neoplastic effect Effects 0.000 abstract description 2
- 238000002474 experimental method Methods 0.000 abstract description 2
- OKKJLVBELUTLKV-UHFFFAOYSA-N Methanol Chemical compound OC OKKJLVBELUTLKV-UHFFFAOYSA-N 0.000 description 27
- CSCPPACGZOOCGX-UHFFFAOYSA-N Acetone Chemical compound CC(C)=O CSCPPACGZOOCGX-UHFFFAOYSA-N 0.000 description 18
- YMWUJEATGCHHMB-UHFFFAOYSA-N Dichloromethane Chemical compound ClCCl YMWUJEATGCHHMB-UHFFFAOYSA-N 0.000 description 12
- XEKOWRVHYACXOJ-UHFFFAOYSA-N Ethyl acetate Chemical compound CCOC(C)=O XEKOWRVHYACXOJ-UHFFFAOYSA-N 0.000 description 12
- 210000004027 cell Anatomy 0.000 description 12
- 238000000926 separation method Methods 0.000 description 8
- 239000000284 extract Substances 0.000 description 7
- WEVYAHXRMPXWCK-UHFFFAOYSA-N Acetonitrile Chemical compound CC#N WEVYAHXRMPXWCK-UHFFFAOYSA-N 0.000 description 6
- DRYDKQOPVBDZMQ-UHFFFAOYSA-N Secalonic acid A Natural products COC(=O)C12Oc3ccc(c(O)c3C(=O)C1=C(O)CC(C)C2O)c4ccc5OC6(C(O)C(C)CC(=C6C(=O)c5c4O)O)C(=O)OC DRYDKQOPVBDZMQ-UHFFFAOYSA-N 0.000 description 6
- VYPSYNLAJGMNEJ-UHFFFAOYSA-N Silicium dioxide Chemical compound O=[Si]=O VYPSYNLAJGMNEJ-UHFFFAOYSA-N 0.000 description 6
- 238000004587 chromatography analysis Methods 0.000 description 6
- 230000002401 inhibitory effect Effects 0.000 description 6
- 239000000741 silica gel Substances 0.000 description 6
- 229910002027 silica gel Inorganic materials 0.000 description 6
- -1 Secalonic acid class compound Chemical class 0.000 description 5
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 description 5
- 239000000287 crude extract Substances 0.000 description 5
- 239000000523 sample Substances 0.000 description 5
- 239000000243 solution Substances 0.000 description 5
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 5
- HEDRZPFGACZZDS-UHFFFAOYSA-N Chloroform Chemical compound ClC(Cl)Cl HEDRZPFGACZZDS-UHFFFAOYSA-N 0.000 description 4
- RTZKZFJDLAIYFH-UHFFFAOYSA-N Diethyl ether Chemical compound CCOCC RTZKZFJDLAIYFH-UHFFFAOYSA-N 0.000 description 4
- 239000003795 chemical substances by application Substances 0.000 description 4
- 238000010828 elution Methods 0.000 description 4
- 235000015097 nutrients Nutrition 0.000 description 4
- 238000000746 purification Methods 0.000 description 4
- 210000004881 tumor cell Anatomy 0.000 description 4
- IAZDPXIOMUYVGZ-UHFFFAOYSA-N Dimethylsulphoxide Chemical compound CS(C)=O IAZDPXIOMUYVGZ-UHFFFAOYSA-N 0.000 description 3
- 229930183845 Secalonic acid Natural products 0.000 description 3
- 238000012360 testing method Methods 0.000 description 3
- OWEGMIWEEQEYGQ-UHFFFAOYSA-N 100676-05-9 Natural products OC1C(O)C(O)C(CO)OC1OCC1C(O)C(O)C(O)C(OC2C(OC(O)C(O)C2O)CO)O1 OWEGMIWEEQEYGQ-UHFFFAOYSA-N 0.000 description 2
- 238000001644 13C nuclear magnetic resonance spectroscopy Methods 0.000 description 2
- FBPFZTCFMRRESA-KVTDHHQDSA-N D-Mannitol Chemical compound OC[C@@H](O)[C@@H](O)[C@H](O)[C@H](O)CO FBPFZTCFMRRESA-KVTDHHQDSA-N 0.000 description 2
- 241000233866 Fungi Species 0.000 description 2
- 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 description 2
- 239000007836 KH2PO4 Substances 0.000 description 2
- GUBGYTABKSRVRQ-PICCSMPSSA-N Maltose Natural products O[C@@H]1[C@@H](O)[C@H](O)[C@@H](CO)O[C@@H]1O[C@@H]1[C@@H](CO)OC(O)[C@H](O)[C@H]1O GUBGYTABKSRVRQ-PICCSMPSSA-N 0.000 description 2
- 229930195725 Mannitol Natural products 0.000 description 2
- 206010028980 Neoplasm Diseases 0.000 description 2
- NFZJAYYORNVZNI-ZKHWAINJSA-N Secalonic acid A Chemical compound O[C@H]1[C@H](C)CC(=O)C2=C(O)C3=C(O)C(C4=CC=C5O[C@]6(C(=C(O)C5=C4O)C(=O)C[C@@H](C)[C@@H]6O)C(=O)OC)=CC=C3O[C@@]21C(=O)OC NFZJAYYORNVZNI-ZKHWAINJSA-N 0.000 description 2
- NFZJAYYORNVZNI-OCHURCMPSA-N Secalonic acid D Chemical compound O[C@@H]1[C@@H](C)CC(=O)C2=C(O)C3=C(O)C(C4=CC=C5O[C@@]6(C(=C(O)C5=C4O)C(=O)C[C@H](C)[C@H]6O)C(=O)OC)=CC=C3O[C@]21C(=O)OC NFZJAYYORNVZNI-OCHURCMPSA-N 0.000 description 2
- MZZSDCJQCLYLLL-UHFFFAOYSA-N Secalonsaeure A Natural products COC(=O)C12OC3C(CC1=C(O)CC(C)C2O)C(=CC=C3c4ccc(O)c5C(=O)C6=C(O)CC(C)C(O)C6(Oc45)C(=O)OC)O MZZSDCJQCLYLLL-UHFFFAOYSA-N 0.000 description 2
- MCWOXLPZYFOWRX-UHFFFAOYSA-N Stemphyperylenol Natural products OC1CC(=O)C2=C(O)C=CC3=C2C1C1=C2C3C(O)CC(=O)C2=C(O)C=C1 MCWOXLPZYFOWRX-UHFFFAOYSA-N 0.000 description 2
- 238000004458 analytical method Methods 0.000 description 2
- 201000011510 cancer Diseases 0.000 description 2
- 229940041514 candida albicans extract Drugs 0.000 description 2
- NEHMKBQYUWJMIP-UHFFFAOYSA-N chloromethane Chemical class ClC NEHMKBQYUWJMIP-UHFFFAOYSA-N 0.000 description 2
- 238000004440 column chromatography Methods 0.000 description 2
- 238000007796 conventional method Methods 0.000 description 2
- 238000005100 correlation spectroscopy Methods 0.000 description 2
- 239000003480 eluent Substances 0.000 description 2
- 229910052564 epsomite Inorganic materials 0.000 description 2
- 238000001914 filtration Methods 0.000 description 2
- 239000012530 fluid Substances 0.000 description 2
- 238000001641 gel filtration chromatography Methods 0.000 description 2
- 238000003919 heteronuclear multiple bond coherence Methods 0.000 description 2
- 230000005764 inhibitory process Effects 0.000 description 2
- 239000007788 liquid Substances 0.000 description 2
- 239000000594 mannitol Substances 0.000 description 2
- 235000010355 mannitol Nutrition 0.000 description 2
- XELZGAJCZANUQH-UHFFFAOYSA-N methyl 1-acetylthieno[3,2-c]pyrazole-5-carboxylate Chemical compound CC(=O)N1N=CC2=C1C=C(C(=O)OC)S2 XELZGAJCZANUQH-UHFFFAOYSA-N 0.000 description 2
- 244000005700 microbiome Species 0.000 description 2
- 229910000402 monopotassium phosphate Inorganic materials 0.000 description 2
- LPUQAYUQRXPFSQ-DFWYDOINSA-M monosodium L-glutamate Chemical compound [Na+].[O-]C(=O)[C@@H](N)CCC(O)=O LPUQAYUQRXPFSQ-DFWYDOINSA-M 0.000 description 2
- 235000013923 monosodium glutamate Nutrition 0.000 description 2
- 239000004223 monosodium glutamate Substances 0.000 description 2
- 239000003208 petroleum Substances 0.000 description 2
- 238000004262 preparative liquid chromatography Methods 0.000 description 2
- 239000011780 sodium chloride Substances 0.000 description 2
- 238000002604 ultrasonography Methods 0.000 description 2
- 239000012138 yeast extract Substances 0.000 description 2
- 238000005160 1H NMR spectroscopy Methods 0.000 description 1
- 241000894006 Bacteria Species 0.000 description 1
- 239000006144 Dulbecco’s modified Eagle's medium Substances 0.000 description 1
- 239000007864 aqueous solution Substances 0.000 description 1
- 238000013096 assay test Methods 0.000 description 1
- 230000008033 biological extinction Effects 0.000 description 1
- 201000001531 bladder carcinoma Diseases 0.000 description 1
- 239000012490 blank solution Substances 0.000 description 1
- 238000004364 calculation method Methods 0.000 description 1
- 230000004663 cell proliferation Effects 0.000 description 1
- 150000001793 charged compounds Chemical class 0.000 description 1
- 239000006071 cream Substances 0.000 description 1
- 238000002425 crystallisation Methods 0.000 description 1
- 230000008025 crystallization Effects 0.000 description 1
- 239000000824 cytostatic agent Substances 0.000 description 1
- 230000001085 cytostatic effect Effects 0.000 description 1
- 230000000994 depressogenic effect Effects 0.000 description 1
- 229940079593 drug Drugs 0.000 description 1
- 238000000119 electrospray ionisation mass spectrum Methods 0.000 description 1
- 238000005516 engineering process Methods 0.000 description 1
- 239000008103 glucose Substances 0.000 description 1
- 238000000338 in vitro Methods 0.000 description 1
- 239000006101 laboratory sample Substances 0.000 description 1
- 239000006193 liquid solution Substances 0.000 description 1
- 239000002609 medium Substances 0.000 description 1
- 238000000034 method Methods 0.000 description 1
- NFZJAYYORNVZNI-UHFFFAOYSA-N methyl 4,8,9-trihydroxy-3-methyl-1-oxo-7-(1,5,9-trihydroxy-10a-methoxycarbonyl-6-methyl-8-oxo-6,7-dihydro-5h-xanthen-2-yl)-3,4-dihydro-2h-xanthene-4a-carboxylate Chemical compound OC1C(C)CC(=O)C2=C(O)C3=C(O)C(C4=CC=C5OC6(C(=C(O)C5=C4O)C(=O)CC(C)C6O)C(=O)OC)=CC=C3OC21C(=O)OC NFZJAYYORNVZNI-UHFFFAOYSA-N 0.000 description 1
- 239000000203 mixture Substances 0.000 description 1
- 230000035479 physiological effects, processes and functions Effects 0.000 description 1
- 239000002504 physiological saline solution Substances 0.000 description 1
- 239000000049 pigment Substances 0.000 description 1
- GNSKLFRGEWLPPA-UHFFFAOYSA-M potassium dihydrogen phosphate Chemical compound [K+].OP(O)([O-])=O GNSKLFRGEWLPPA-UHFFFAOYSA-M 0.000 description 1
- 238000012545 processing Methods 0.000 description 1
- 239000012488 sample solution Substances 0.000 description 1
- 230000024053 secondary metabolic process Effects 0.000 description 1
- 239000007787 solid Substances 0.000 description 1
- 238000001228 spectrum Methods 0.000 description 1
- 230000003068 static effect Effects 0.000 description 1
- 239000000126 substance Substances 0.000 description 1
- 239000006228 supernatant Substances 0.000 description 1
- 238000010998 test method Methods 0.000 description 1
- 125000003831 tetrazolyl group Chemical group 0.000 description 1
- 210000003932 urinary bladder Anatomy 0.000 description 1
- 208000010570 urinary bladder carcinoma Diseases 0.000 description 1
- 150000007964 xanthones Chemical class 0.000 description 1
Classifications
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07D—HETEROCYCLIC COMPOUNDS
- C07D311/00—Heterocyclic compounds containing six-membered rings having one oxygen atom as the only hetero atom, condensed with other rings
- C07D311/02—Heterocyclic compounds containing six-membered rings having one oxygen atom as the only hetero atom, condensed with other rings ortho- or peri-condensed with carbocyclic rings or ring systems
- C07D311/78—Ring systems having three or more relevant rings
- C07D311/80—Dibenzopyrans; Hydrogenated dibenzopyrans
- C07D311/82—Xanthenes
- C07D311/84—Xanthenes with hetero atoms or with carbon atoms having three bonds to hetero atoms with at the most one bond to halogen, e.g. ester or nitrile radicals, directly attached in position 9
- C07D311/86—Oxygen atoms, e.g. xanthones
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12P—FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
- C12P17/00—Preparation of heterocyclic carbon compounds with only O, N, S, Se or Te as ring hetero atoms
- C12P17/02—Oxygen as only ring hetero atoms
- C12P17/06—Oxygen as only ring hetero atoms containing a six-membered hetero ring, e.g. fluorescein
Landscapes
- Organic Chemistry (AREA)
- Chemical & Material Sciences (AREA)
- Engineering & Computer Science (AREA)
- Zoology (AREA)
- Life Sciences & Earth Sciences (AREA)
- Wood Science & Technology (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Microbiology (AREA)
- General Chemical & Material Sciences (AREA)
- Biotechnology (AREA)
- Health & Medical Sciences (AREA)
- Biochemistry (AREA)
- Bioinformatics & Cheminformatics (AREA)
- General Engineering & Computer Science (AREA)
- General Health & Medical Sciences (AREA)
- Genetics & Genomics (AREA)
- Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
- Preparation Of Compounds By Using Micro-Organisms (AREA)
- Medicines Containing Plant Substances (AREA)
Abstract
The present invention relates to come from the secalonic acid H of penicillium oxalicum and prepare the application of anti-human colon cancer drug.The compound, which has, suppresses Proliferation of Human Colon effect.Its structural formula is：.By fermented and cultured penicillium oxalicum (Penicillium oxalicum) IBPT 6, fermentate is obtained, the compound is then isolated and purified out from fermentate.It is verified by experiments, the compound has preferable antitumor activity to human colon cancer cell HCT116.The research of medicine or antineoplastic for human colon carcinoma can be suppressed as Proliferation of Human Colon is prepared.
Description
Technical field
The present invention relates to coming from the secalonic acid H of penicillium oxalicum and preparing the application of anti-human colon cancer drug, belong to medicine
Field.
Background technology
Secalonic acid class compound（Secalonic acids）Belong to ergot pigment（Ergochrome）Secondary metabolism is produced
Thing, is xanthone dimer.Since Stoll etc. was in the isolated secalonic acid A from fungi in 1952
（Secalonic acid A）Afterwards, the serial compound secalonic acid（A, B, C, D, E, F, G）Just constantly by
It was found that and research.Secalonic acid class compound has a variety of physiologically actives, with secalonic acid D（Secalonic acid
D, SAD）Exemplified by, 5 mg/ml SAD is added in physiological saline, in the range of 5-20 mg, you can to treat early stage bladder
In the range of cancer, 50-100 mg, and be free from side effects generation effective in cure to more serious carcinoma of urinary bladder.It has been investigated that, some seas
Foreign fungi can produce the secalonic acid class compound that structure is novel, activity is good during cometabolism, with good medicine
With and industrialization prospect.
The present inventor studies and learnt, penicillium oxalicum (Penicillium oxalicum) IBPT-6, (in 2013
December 25 was deposited in China typical culture collection center, address:Wuhan Wuhan University, deposit number is：CCTCC NO：
M 2013714) the crude extract of tunning have good cell inhibitory effect activity, its active component is ground then
Study carefully.Research finds that shown secalonic acid class compound has anti-human colon cancer reactive, has not yet to see the compound to people's colon
The report of the proliferation inhibition activity of cancer cell, therefore in the market is also there is not yet medicine related to this.
The content of the invention
It is an object of the invention to provide the secalonic acid H for coming from penicillium oxalicum and prepare anti-human colon cancer drug should
With.The compound, which has, suppresses Colon Cancer Cells effect, with anti-human colon cancer reactive.Its structural formula is：
。
The preparation method of the compound, be by fermented and cultured penicillium oxalicum (Penicillium oxalicum)
IBPT-6, obtains fermentate, the compound is then isolated and purified out from fermentate.Comprise the following steps that：
1 fermenting and producing
Cultivate microorganism conventional method, take penicillium oxalicum (Penicillium oxalicum) IBPT-6 be inoculated into PDA consolidate
Cultivate 2 to 3 days, be then seeded into nutrient solution in 28 DEG C of incubators on body slant medium, 28 DEG C of static gas wave refrigerators are after 30 days,
Obtain mycelium and zymotic fluid；The nutrient solution composition：Every liter of water contains the g of mannitol 20.0, the g of yeast extract 3.0, maltose 20.0
G, the g of monosodium glutamate 10.0, glucose 10.0 g, KH2PO4 0.5 g、MgSO4·7H2O 0.3 g、NaCl 15.0 g；
The acquisition of 2 medicinal extract
With gauze by mycelium and separation of fermentative broth.Mycelium acetone soln（Containing 20% ~ 30% water）Continuous ultrasound broken wall 3 times,
Filtering removes residue, obtains the mycelial crude extract containing acetone and water.Be concentrated under reduced pressure removal acetone, obtains the water-soluble of runic thing
Liquid, then with volume ratio 1:2, which add ethyl acetate, extracts 3 times, obtains ethyl acetate crude extract, is concentrated under reduced pressure near dry that mycelium soaks
The g of cream 36.5.
The separation and purification of 3 compounds
Mycelium medicinal extract is by 100-200 mesh silica gel mixed samples, with petroleum ether：Dichloromethane：Methanol is gradient eluent, is subtracted
Press silica gel chromatograph column chromatography.By simple thin-layer chromatographic analysis, merge, be separated into component A-E.Component D (5.9 g) (two
Chloromethanes：Methanol v/v=100:1 eluate) with dichloromethane：Methanol is gradient elution agent, carries out pressured column silica gel
Analysis, merges after thin-layer chromatographic analysis and obtains five subfraction D1-D5.Component D2 (1.2 g) is with chloroform：Methanol=
1:2 be gradient elution agent, carries out gel filtration chromatography（Sephadex LH-20）, merge after thin-layer chromatographic analysis and obtain four
Subfraction D2-1 ~ D2-4.D2 subfraction D2-3 (212 mg) by semi-preparative liquid chromatography (1010 type ODS-A, 10 ×
250 mm, 5 μm)：Separation flow velocity is 5 mL/min, and mobile phase is that 55% acetonitrile contains 0.1% TFA, obtains shown compound (2.4
Mg, tR 8.1 min)。
The penicillium oxalicum (Penicillium oxalicum) IBPT-6, on December 25th, 2013 is deposited in
State's Type Tissue Collection, address:Wuhan Wuhan University, deposit number is：CCTCC NO：M 2013714.
The present invention also protects described compound preparing the application in suppressing Proliferation of Human Colon medicine, and should
Application of the compound in anti-human colon cancer drug is prepared.
The remarkable advantage of the present invention：The secalonic acid compound shown in research has no report and suppresses people's knot with significant
Colon-cancer cell proliferation activity, has not yet to see report of the compound to Proliferation of Human Colon inhibitory activity, therefore market
On also there is not yet medicine related to this.
Brief description of the drawings
COSY, HMBC and NOE signal main Fig. 1 Secalonic acid H.
Embodiment
The chemical constitution of signified compound in examples below：
The fermenting and producing and separation and purification of the compound of embodiment 1
1 fermenting and producing
Produce the fermented and cultured of bacterium：By culture microorganism conventional method, take penicillium oxalicum (Penicillium oxalicum)
IBPT-6 (is deposited in China typical culture collection center, address on December 25th, 2013:Wuhan Wuhan University, protects
Hiding numbering is：CCTCC NO：M 2013714) in right amount, it is inoculated into PDA solid slope culture mediums and is cultivated in 28 DEG C of incubators
3 days.
Take the inclined-plane culture penicillium oxalicum of 2 to 3 days (Penicillium oxalicum) appropriate IBPT-6, it is inoculated into dress
By 400mL nutrient solutions, [nutrient solution is constituted (g/l)：Mannitol 20.0, yeast extract 3.0, maltose 20.0, monosodium glutamate 10.0, Portugal
Grape sugar 10.0, KH2PO40.5, MgSO4·7H2O 0.3, NaCl 15.0, constant volume] 1000mL conical flasks in, 28 DEG C are quiet
After only cultivating 30 days, mycelium and zymotic fluid are obtained.
The acquisition of 2 medicinal extract
With gauze by mycelium and separation of fermentative broth.Mycelium acetone soln（Containing 30% water）Continuous ultrasound broken wall 3 times, filtering
Residue is removed, the mycelial crude extract containing acetone and water is obtained.Be concentrated under reduced pressure removal acetone, obtains the aqueous solution of runic thing,
Again with volume ratio 1:2, which add ethyl acetate, extracts 3 times, obtains ethyl acetate crude extract, is concentrated under reduced pressure into and closely does to obtain mycelium medicinal extract
36.5 g。
The separation and purification of 3 compounds
Mycelium medicinal extract is by 100-200 mesh silica gel mixed samples, with petroleum ether：Dichloromethane：Methanol is gradient eluent, is subtracted
Press silica gel chromatograph column chromatography.By simple thin-layer chromatographic analysis, merge, be separated into component A-E.Component D (5.9 g) (two
Chloromethanes：Methanol v/v=100:1 eluate) with dichloromethane：Methanol is gradient elution agent, carries out pressured column silica gel
Analysis, merges after thin-layer chromatographic analysis and obtains five subfraction D1-D5.Component D2 (1.2 g) is with chloroform：Methanol=
1:2 be gradient elution agent, carries out gel filtration chromatography（Sephadex LH-20）, merge after thin-layer chromatographic analysis and obtain four
Subfraction D2-1 ~ D2-4.D2 subfraction D2-3 (212 mg) by semi-preparative liquid chromatography (1010 type ODS-A, 10 ×
250 mm, 5 μm)：Separation flow velocity is 5 mL/min, and mobile phase is that 55% acetonitrile contains 0.1% TFA, obtains shown compound (2.4
Mg, tR 8.1 min)。
It is yellow oil under compound normal temperature, high-resolution electrospray ionization mass spectrum HRESI-MS existsm/z：Given at 659.1370
Go out molecular ion peak [M+Na]+（calcd for C32H28NaO14, 659.1377）；It is 636 to point out molecular weight, with reference to wave spectrum letter
Breath speculates that molecular formula is C32H28O14。1H and13C-NMR data are shown in Table 1, and main COSY, HMBC and NOE signal is shown in Fig. 1.
The compound of table 11H and13C-NMR data (500 MHz1H and 126 MHz 13C, in DMSO-d 6 )
The test of the anti tumor activity in vitro of embodiment 2
1 laboratory sample and experimental method
The preparation test sample of sample solution is the pure compounds of separation and purification in above-mentioned implementation 1.Precision is weighed in right amount
Sample, the solution of concentration needed for being configured to methanol, for surveying activity.
The squamous subculture of cell line and cell uses tumor cell line, and tumour cell uses the DMEM containing 10% FBS to cultivate
Base, at 37 DEG C in being passed through 5% CO2Incubator in squamous subculture.
Cell inhibitory effect activity test method
Tetrazolium（MTT）Method is taken the logarithm the tumour cell in growth period, and cell density is adjusted into every milliliter 1 × 105Individual cell,
It is inoculated in by every 200 microlitres of hole in 96 porocyte culture plates, 5% CO is passed through in 37 DEG C2Incubator in cultivate 4 hours.Per hole
The sample liquid or blank solution of 2 microlitres of addition, after cultivating 24 hours, MTT liquid is added per hole（MTT every milliliter of 5 milligrams of physiology
Saline solution）10 microlitres, continue to cultivate 4 hours, 37 DEG C, 2000 revs/min centrifuge 8 minutes, draw supernatant.DMSO is added per hole
Each 100 microlitres, vibrate 15 minutes, after being completely dissolved to crystallization, produced using MD companies on micro oscillator
SPECTRAMAX Plus types ELIASA is determined per extinction of the hole at 570 nm（OD）Value.The sample in the orifice plate of same 96
Each concentration is respectively provided with three holes, the another blank control for setting three holes and acellular withered hole（If medicine has color to do accordingly
It is withered that drug concentration is acellular）.Each hole OD values first do corresponding acellular withered, then take three hole mean OD values by IR (%)=
(ODBlank control-ODSample)/ODBlank control× 100% calculates the proliferation inhibition rate of cell under each concentration（IR%）.
2. experimental result
Cell inhibitory effect active testing result
In mtt assay test, according to the Cytostatic to tumor cell rate of the compound of various concentrations, using SPSS16.0 softwares
Carry out data processing and calculation of half inhibitory concentration IC50Value.It the results are shown in Table 2.
Inhibitory activity of the compound of table 2 to Proliferation of Human Colon
3. conclusion
The compound has preferable antitumor activity to human colon cancer cell.Can be as preparing Colon Cancer Cells depressant
Thing or antineoplastic are used for the research of colon cancer.
Claims (4)
1. compound。
2. the preparation method of compound as claimed in claim 1, it is characterised in that：Fermented and cultured penicillium oxalicum
(Penicillium oxalicum) IBPT-6, fermentate is obtained, the compound, institute are then isolated and purified out from fermentate
State penicillium oxalicum (Penicillium oxalicum) IBPT-6, it is deposited in Chinese Typical Representative culture on December 25th, 2013
Thing collection, address:Wuhan Wuhan University, deposit number is：CCTCC NO：M 2013714.
3. the compound described in claim 1 is preparing the application in suppressing Proliferation of Human Colon medicine.
4. application of the compound in anti-human colon cancer drug is prepared described in claim 1.
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| CN109106702A (en) * | 2017-12-19 | 2019-01-01 | 福州大学 | Derived from application of 4-4 ' the isomerization secalonic acid D in terms of colon cancer of penicillium oxalicum |
| CN109106706A (en) * | 2017-12-19 | 2019-01-01 | 福州大学 | Derived from application of 4-4 ' the isomerization secalonic acid D in terms of gastric cancer of penicillium oxalicum |
| CN109776478A (en) * | 2018-12-04 | 2019-05-21 | 福州大学 | Iso-Penicillixanthone A derived from penicillium oxalicum and the application in terms of cervical carcinoma |
| CN109776477A (en) * | 2018-12-04 | 2019-05-21 | 福建省肿瘤医院（福建省肿瘤研究所、福建省癌症防治中心） | Iso-Penicillixanthone A and anti-vincristine drug resistance application derived from penicillium oxalicum |
| CN110407792A (en) * | 2019-04-26 | 2019-11-05 | 福州大学 | Derived from the secalonic acid class compound Secalonic acid J and preparation method of penicillium oxalicum |
| CN110407794A (en) * | 2019-04-26 | 2019-11-05 | 福州大学 | Secalonic acid K derived from penicillium oxalicum and the application on inhibition cancer cell multiplication |
| CN110627763A (en) * | 2018-12-04 | 2019-12-31 | 福建省肿瘤医院（福建省肿瘤研究所、福建省癌症防治中心） | iso-Penicillium xanthone A from Penicillium oxalicum and application of anti-cisplatin drug resistance |
| CN110669800A (en) * | 2018-12-04 | 2020-01-10 | 福建省肿瘤医院（福建省肿瘤研究所、福建省癌症防治中心） | Application of iso-Penicillium xanthone A from Penicillium oxalicum and adriamycin resistance |
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| CN110917185A (en) * | 2018-12-04 | 2020-03-27 | 福州大学 | iso-Penicillium xanthone A from penicillium oxalicum and application in aspect of breast cancer |
| CN110922378A (en) * | 2018-12-04 | 2020-03-27 | 福州大学 | iso-Penicillium xanthone A from penicillium oxalicum and application in nasopharyngeal carcinoma |
| CN110922377A (en) * | 2018-12-04 | 2020-03-27 | 福州大学 | Application of iso-Penicillium xanthone A from Penicillium oxalicum in aspect of melanoma |
| CN110923279A (en) * | 2018-12-04 | 2020-03-27 | 福州大学 | iso-Penicillium xanthone A from penicillium oxalicum and application in colon cancer |
| CN110922383A (en) * | 2018-12-04 | 2020-03-27 | 福州大学 | iso-Penicillium xanthone A from Penicillium oxalicum and its application in gastric cancer |
| CN110922381A (en) * | 2018-12-04 | 2020-03-27 | 福州大学 | iso-Penicillium xanthone A from penicillium oxalicum and application in esophagus cancer |
| CN110922380A (en) * | 2018-12-04 | 2020-03-27 | 福州大学 | iso-Penicillium xanthone A from Penicillium oxalicum and its application in liver cancer |
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Citations (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JPS5529912A (en) * | 1978-08-21 | 1980-03-03 | Asahi Chem Ind Co Ltd | Production of secalonic acid by microorganism |
| JPS57171989A (en) * | 1981-04-17 | 1982-10-22 | Asahi Chem Ind Co Ltd | Aminometyl-secalonic acid and its preparation |
| CN102408997A (en) * | 2010-11-29 | 2012-04-11 | 国家海洋局第三海洋研究所 | Deep-sea-sourced penicillium F11 capable of producing compound secalonic acid F with cytotoxic activity |
| CN104611389A (en) * | 2014-12-10 | 2015-05-13 | 沈阳药科大学 | Fermentation optimizing technology for producing secalonic acid D (SAD) by using P.oxalicum |
| CN107298670A (en) * | 2017-06-17 | 2017-10-27 | 福州大学 | Come from penicillium oxalicum secalonic acid H and prepare anti-human oral cavity epidermoid carcinoma medicinal application |
| CN107485607A (en) * | 2017-06-17 | 2017-12-19 | 福州大学 | The secalonic acid H for coming from penicillium oxalicum is preparing the application of anti-human oesophagus cancer drug |
| CN109134416A (en) * | 2017-06-17 | 2019-01-04 | 福州大学 | Secalonic acid H derived from penicillium oxalicum is in the application for preparing human cervical cancer 1 cancer drug |
-
2017
- 2017-06-17 CN CN201710460067.7A patent/CN107298671B/en not_active Expired - Fee Related
Patent Citations (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JPS5529912A (en) * | 1978-08-21 | 1980-03-03 | Asahi Chem Ind Co Ltd | Production of secalonic acid by microorganism |
| JPS57171989A (en) * | 1981-04-17 | 1982-10-22 | Asahi Chem Ind Co Ltd | Aminometyl-secalonic acid and its preparation |
| CN102408997A (en) * | 2010-11-29 | 2012-04-11 | 国家海洋局第三海洋研究所 | Deep-sea-sourced penicillium F11 capable of producing compound secalonic acid F with cytotoxic activity |
| CN104611389A (en) * | 2014-12-10 | 2015-05-13 | 沈阳药科大学 | Fermentation optimizing technology for producing secalonic acid D (SAD) by using P.oxalicum |
| CN107298670A (en) * | 2017-06-17 | 2017-10-27 | 福州大学 | Come from penicillium oxalicum secalonic acid H and prepare anti-human oral cavity epidermoid carcinoma medicinal application |
| CN107485607A (en) * | 2017-06-17 | 2017-12-19 | 福州大学 | The secalonic acid H for coming from penicillium oxalicum is preparing the application of anti-human oesophagus cancer drug |
| CN109134416A (en) * | 2017-06-17 | 2019-01-04 | 福州大学 | Secalonic acid H derived from penicillium oxalicum is in the application for preparing human cervical cancer 1 cancer drug |
Non-Patent Citations (2)
| Title |
|---|
| GANG CHEN ET AL.: "Isolation, Structure Determination, In Vivo/Vitro Assay and Docking Study of a Xanthone with antitumor activity from Fungus Penicillium oxalicum", 《REC. NAT. PROD.》 * |
| 徐帮等: "中华剑角蝗共生真菌Penicillium oxalicum中苯并吡喃酮二聚体类化合物的研究", 《天然产物研究与开发》 * |
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| CN109106702A (en) * | 2017-12-19 | 2019-01-01 | 福州大学 | Derived from application of 4-4 ' the isomerization secalonic acid D in terms of colon cancer of penicillium oxalicum |
| CN109106706A (en) * | 2017-12-19 | 2019-01-01 | 福州大学 | Derived from application of 4-4 ' the isomerization secalonic acid D in terms of gastric cancer of penicillium oxalicum |
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| CN110922380A (en) * | 2018-12-04 | 2020-03-27 | 福州大学 | iso-Penicillium xanthone A from Penicillium oxalicum and its application in liver cancer |
| CN110923279A (en) * | 2018-12-04 | 2020-03-27 | 福州大学 | iso-Penicillium xanthone A from penicillium oxalicum and application in colon cancer |
| CN109776478B (en) * | 2018-12-04 | 2022-10-14 | 福州大学 | iso-Penicillium xanthone A from penicillium oxalicum and application in cervical cancer |
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| CN110917185A (en) * | 2018-12-04 | 2020-03-27 | 福州大学 | iso-Penicillium xanthone A from penicillium oxalicum and application in aspect of breast cancer |
| CN110922378A (en) * | 2018-12-04 | 2020-03-27 | 福州大学 | iso-Penicillium xanthone A from penicillium oxalicum and application in nasopharyngeal carcinoma |
| CN110922377A (en) * | 2018-12-04 | 2020-03-27 | 福州大学 | Application of iso-Penicillium xanthone A from Penicillium oxalicum in aspect of melanoma |
| CN109776478A (en) * | 2018-12-04 | 2019-05-21 | 福州大学 | Iso-Penicillixanthone A derived from penicillium oxalicum and the application in terms of cervical carcinoma |
| CN110922383A (en) * | 2018-12-04 | 2020-03-27 | 福州大学 | iso-Penicillium xanthone A from Penicillium oxalicum and its application in gastric cancer |
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| CN109776477A (en) * | 2018-12-04 | 2019-05-21 | 福建省肿瘤医院（福建省肿瘤研究所、福建省癌症防治中心） | Iso-Penicillixanthone A and anti-vincristine drug resistance application derived from penicillium oxalicum |
| CN110407792A (en) * | 2019-04-26 | 2019-11-05 | 福州大学 | Derived from the secalonic acid class compound Secalonic acid J and preparation method of penicillium oxalicum |
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| CN110407794B (en) * | 2019-04-26 | 2022-10-14 | 福州大学 | Selenolonic acid K derived from penicillium oxalicum and application thereof in inhibiting cancer cell proliferation |
| CN110407792B (en) * | 2019-04-26 | 2022-10-14 | 福州大学 | Secalonic acid J compound derived from penicillium oxalicum and preparation method thereof |
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