CN110407793A - 源于草酸青霉的黑麦酮酸j及在抑制癌细胞增殖上的应用 - Google Patents
源于草酸青霉的黑麦酮酸j及在抑制癌细胞增殖上的应用 Download PDFInfo
- Publication number
- CN110407793A CN110407793A CN201910344814.XA CN201910344814A CN110407793A CN 110407793 A CN110407793 A CN 110407793A CN 201910344814 A CN201910344814 A CN 201910344814A CN 110407793 A CN110407793 A CN 110407793A
- Authority
- CN
- China
- Prior art keywords
- cancer cell
- penicillium oxalicum
- compound
- application
- derived
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Granted
Links
- 206010028980 Neoplasm Diseases 0.000 title claims abstract description 13
- 201000011510 cancer Diseases 0.000 title claims abstract description 13
- 241000985513 Penicillium oxalicum Species 0.000 title abstract description 19
- WIZZCXFBFFCTSY-RYGOELPRSA-N C[C@H]1CC(=O)C2=C(C3=C(C=CC(=C3O)C4=C(C5=C(C=C4C)O[C@]6([C@@H](CCC(=O)C6=C5O)O)C(=O)OC)O)O[C@]2([C@@H]1O)C(=O)OC)O Chemical compound C[C@H]1CC(=O)C2=C(C3=C(C=CC(=C3O)C4=C(C5=C(C=C4C)O[C@]6([C@@H](CCC(=O)C6=C5O)O)C(=O)OC)O)O[C@]2([C@@H]1O)C(=O)OC)O WIZZCXFBFFCTSY-RYGOELPRSA-N 0.000 title abstract description 4
- 230000005764 inhibitory process Effects 0.000 title description 5
- 150000001875 compounds Chemical class 0.000 claims abstract description 27
- 239000003795 chemical substances by application Substances 0.000 claims description 6
- 238000002360 preparation method Methods 0.000 claims description 4
- 239000003560 cancer drug Substances 0.000 claims description 2
- 230000000694 effects Effects 0.000 abstract description 8
- 230000002401 inhibitory effect Effects 0.000 abstract description 8
- 239000003814 drug Substances 0.000 abstract description 6
- 229940079593 drug Drugs 0.000 abstract description 6
- 238000000855 fermentation Methods 0.000 abstract description 6
- 230000004151 fermentation Effects 0.000 abstract description 6
- 230000009702 cancer cell proliferation Effects 0.000 abstract description 4
- 239000000463 material Substances 0.000 abstract description 4
- 239000002246 antineoplastic agent Substances 0.000 abstract description 2
- 229940041181 antineoplastic drug Drugs 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
- 210000004027 cell Anatomy 0.000 description 15
- 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
- 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 8
- VYPSYNLAJGMNEJ-UHFFFAOYSA-N Silicium dioxide Chemical compound O=[Si]=O VYPSYNLAJGMNEJ-UHFFFAOYSA-N 0.000 description 8
- DTQVDTLACAAQTR-UHFFFAOYSA-N Trifluoroacetic acid Chemical compound OC(=O)C(F)(F)F DTQVDTLACAAQTR-UHFFFAOYSA-N 0.000 description 8
- 239000000741 silica gel Substances 0.000 description 8
- 229910002027 silica gel Inorganic materials 0.000 description 8
- 239000000287 crude extract Substances 0.000 description 7
- 239000000523 sample Substances 0.000 description 7
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 7
- WEVYAHXRMPXWCK-UHFFFAOYSA-N Acetonitrile Chemical compound CC#N WEVYAHXRMPXWCK-UHFFFAOYSA-N 0.000 description 6
- IAZDPXIOMUYVGZ-UHFFFAOYSA-N Dimethylsulphoxide Chemical compound CS(C)=O IAZDPXIOMUYVGZ-UHFFFAOYSA-N 0.000 description 6
- 238000000926 separation method Methods 0.000 description 6
- 229930183845 Secalonic acid Natural products 0.000 description 5
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 description 5
- 239000007788 liquid Substances 0.000 description 5
- 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 class 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 5
- 239000000243 solution Substances 0.000 description 5
- RTZKZFJDLAIYFH-UHFFFAOYSA-N Diethyl ether Chemical compound CCOCC RTZKZFJDLAIYFH-UHFFFAOYSA-N 0.000 description 4
- -1 Secalonic acid class compound Chemical class 0.000 description 4
- 238000004587 chromatography analysis Methods 0.000 description 4
- 238000010828 elution Methods 0.000 description 4
- 239000000284 extract Substances 0.000 description 4
- 238000000746 purification Methods 0.000 description 4
- 210000004881 tumor cell Anatomy 0.000 description 4
- 230000035755 proliferation Effects 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
- 206010005003 Bladder cancer Diseases 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
- 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
- 208000007097 Urinary Bladder Neoplasms Diseases 0.000 description 2
- 239000002253 acid Substances 0.000 description 2
- 238000004458 analytical method Methods 0.000 description 2
- 230000001093 anti-cancer Effects 0.000 description 2
- 230000000259 anti-tumor effect Effects 0.000 description 2
- 229940041514 candida albicans extract Drugs 0.000 description 2
- 150000001793 charged compounds Chemical class 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
- 239000006071 cream Substances 0.000 description 2
- 239000003480 eluent Substances 0.000 description 2
- 238000001914 filtration Methods 0.000 description 2
- 239000001963 growth medium Substances 0.000 description 2
- 238000003919 heteronuclear multiple bond coherence Methods 0.000 description 2
- 239000000594 mannitol Substances 0.000 description 2
- 235000010355 mannitol Nutrition 0.000 description 2
- 238000004519 manufacturing process Methods 0.000 description 2
- 238000000034 method Methods 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
- 239000007787 solid Substances 0.000 description 2
- 230000003068 static effect Effects 0.000 description 2
- 239000000126 substance Substances 0.000 description 2
- 238000002604 ultrasonography Methods 0.000 description 2
- 201000005112 urinary bladder cancer Diseases 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
- PEDCQBHIVMGVHV-UHFFFAOYSA-N Glycerine Chemical compound OCC(O)CO PEDCQBHIVMGVHV-UHFFFAOYSA-N 0.000 description 1
- 230000003698 anagen phase Effects 0.000 description 1
- 238000013096 assay test Methods 0.000 description 1
- WQZGKKKJIJFFOK-VFUOTHLCSA-N beta-D-glucose Chemical compound OC[C@H]1O[C@@H](O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-VFUOTHLCSA-N 0.000 description 1
- 230000008033 biological extinction Effects 0.000 description 1
- 239000012490 blank solution Substances 0.000 description 1
- 238000004364 calculation method Methods 0.000 description 1
- 239000000470 constituent 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
- 238000000119 electrospray ionisation mass spectrum Methods 0.000 description 1
- 229910052564 epsomite Inorganic materials 0.000 description 1
- 239000008103 glucose Substances 0.000 description 1
- 238000000338 in vitro Methods 0.000 description 1
- 150000002576 ketones Chemical class 0.000 description 1
- 239000006101 laboratory sample Substances 0.000 description 1
- 239000006193 liquid solution Substances 0.000 description 1
- 229910052943 magnesium sulfate Inorganic materials 0.000 description 1
- CSNNHWWHGAXBCP-UHFFFAOYSA-L magnesium sulphate Substances [Mg+2].[O-][S+2]([O-])([O-])[O-] CSNNHWWHGAXBCP-UHFFFAOYSA-L 0.000 description 1
- 230000004060 metabolic process Effects 0.000 description 1
- 239000000203 mixture Substances 0.000 description 1
- 238000012986 modification Methods 0.000 description 1
- 230000004048 modification Effects 0.000 description 1
- 230000001766 physiological effect Effects 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
- 239000006228 supernatant Substances 0.000 description 1
- 238000010998 test method Methods 0.000 description 1
- 125000003831 tetrazolyl group Chemical group 0.000 description 1
- JNELGWHKGNBSMD-UHFFFAOYSA-N xanthone Chemical group C1=CC=C2C(=O)C3=CC=CC=C3OC2=C1 JNELGWHKGNBSMD-UHFFFAOYSA-N 0.000 description 1
Classifications
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P35/00—Antineoplastic agents
-
- 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
- Chemical & Material Sciences (AREA)
- Organic Chemistry (AREA)
- Life Sciences & Earth Sciences (AREA)
- Health & Medical Sciences (AREA)
- General Health & Medical Sciences (AREA)
- Chemical Kinetics & Catalysis (AREA)
- General Chemical & Material Sciences (AREA)
- Zoology (AREA)
- Wood Science & Technology (AREA)
- Engineering & Computer Science (AREA)
- Veterinary Medicine (AREA)
- Medicinal Chemistry (AREA)
- Animal Behavior & Ethology (AREA)
- Pharmacology & Pharmacy (AREA)
- Biotechnology (AREA)
- Microbiology (AREA)
- Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
- Public Health (AREA)
- Biochemistry (AREA)
- Bioinformatics & Cheminformatics (AREA)
- General Engineering & Computer Science (AREA)
- Genetics & Genomics (AREA)
- Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
- Preparation Of Compounds By Using Micro-Organisms (AREA)
Abstract
本发明涉及一种源于草酸青霉的黑麦酮酸J及在抑制癌细胞增殖上的应用。该化合物具有抑制人癌细胞增殖作用。其结构式为：。通过发酵培养草酸青霉(Penicillium oxalicum)IBPT‑6，获取发酵物，然后从发酵物中分离纯化出该化合物。经实验证实，该化合物对人癌细胞具有较好的活性。可用于制备人癌细胞增殖抑制药物或抗肿瘤药物。
Description
技术领域
本发明涉及一种源于草酸青霉的黑麦酮酸类化合物Secalonic acid J及其抑制人癌细胞增殖方面的应用，属于医药生物领域。
背景技术
黑麦酮酸类化合物（Secalonic acids）属于麦角色素类（Ergochrome）次生代谢产物，为氧杂蒽酮二聚物。自从 Stoll 等在 1952 年从真菌中分离得到黑麦酮酸 A（Secalonic acid A）之后，该系列的化合物黑麦酮酸（A-I）就不断地被发现及研究。黑麦酮酸类化合物具有各种不同的生理活性，以黑麦酮酸D（Secalonic acid D, SAD）为例，5 mg/ml的 SAD加入到生理盐水中, 在5-20 mg范围内，即可以治疗早期膀胱癌，50-100 mg范围内，对更严重的膀胱癌有疗效并且没有副作用发生。经研究发现，一些海洋真菌能够在次级代谢过程中产生结构新颖、活性好的黑麦酮酸类化合物，具有很好的药用和产业化前景。
本发明人研究得知，草酸青霉 (Penicillium oxalicum) IBPT-6 (已于2013年12月25日保藏在中国典型培养物保藏中心，地址: 武汉 武汉大学，保藏编号是：CCTCC NO：M2013714) 发酵产物的粗提取物有很好的细胞增殖抑制活性，遂对其活性成分进行研究。研究发现所示黑麦酮酸类化合物具有抗癌活性，目前尚未见该化合物对癌细胞的增殖抑制活性的报道，因此市场上也尚未见有与此相关的药物。
发明内容
本发明的目的在于提供一种源于草酸青霉的黑麦酮酸类化合物Secalonic acidJ及其抑制人癌细胞增殖方面的应用。该化合物具有抑制癌细胞增殖作用，具有抗癌活性。其结构式为：
。
该化合物的制备方法，是通过发酵培养草酸青霉 (Penicillium oxalicum)IBPT-6，获取发酵物，然后从发酵物中分离纯化出该化合物。具体步骤如下：
1发酵生产
按培养微生物的常规方法，取草酸青霉 (Penicillium oxalicum) IBPT-6接种到PDA固体斜面培养基上在28℃培养箱中培养2至3天，然后接种到培养液中，28℃静止培养30天后，获得菌丝体和发酵液；所述培养液组成：每升水含甘露醇20.0 g、酵母膏3.0 g、麦芽糖20.0 g、味精10.0 g、葡萄糖10.0 g、KH2PO4 0.5 g、MgSO4·7H2O 0.3 g、NaCl 15.0 g。
2 浸膏的获得
用纱布将菌丝体和发酵液分离。菌丝体用丙酮溶液（含20%~30%水）连续超声破壁3次，过滤去除残渣，得到含丙酮和水的菌丝体粗提物。减压浓缩去除丙酮，得到粗提物的水溶液，再以体积比1:2加入乙酸乙酯萃取3次，得乙酸乙酯粗提液，减压浓缩至近干得菌丝体浸膏36.5 g。
3 化合物的分离精制
菌丝体浸膏通过100-200目硅胶拌样，以石油醚：二氯甲烷：甲醇为梯度洗脱液，进行减压硅胶色谱柱层析。经过简单的薄层色谱分析，合并，分离成组分A-E。组分D (5.9 g) (二氯甲烷：甲醇v/v=100:1的洗脱物)以二氯甲烷：甲醇为梯度洗脱剂，进行加压柱硅胶色谱层析，经过薄层色谱分析后合并得到五个亚组分D1-D5。组分D3（1.1 g）以甲醇：酸水（含0.1%三氟乙酸）为梯度洗脱剂进行反向硅胶色谱层析，分离得到四个亚组分D3-1~D3-4。D3的亚组分D3-2（168 mg）通过半制备液相色谱在流动相为50%乙腈（含0.1%三氟乙酸）条件下分离得到所述化合物（2.8 mg）。
所述草酸青霉 (Penicillium oxalicum) IBPT-6，已于2013年12月25日保藏在中国典型培养物保藏中心，地址: 武汉 武汉大学，保藏编号是：CCTCC NO：M 2013714。
本发明还保护了所述的化合物在制备抑制人癌细胞增殖药物中的应用，及该化合物在制备抗人癌药物中的应用。
本发明的显著优点：研究所示该黑麦酮酸化合物未见报道且具有显著的抑制癌细胞增殖活性，目前尚未见该化合物对癌细胞增殖抑制活性的报道，因此市场上也尚未见有与此相关的药物。
附图说明
图1 Secalonic acid J主要的COSY，HMBC和NOE信号。
具体实施方式
在如下的实施例中所指的化合物的化学结构：
实施例1该化合物的发酵生产及分离精制
1发酵生产
生产菌的发酵培养：按培养微生物的常规方法，取草酸青霉 (Penicillium oxalicum)IBPT-6 (已于2013年12月25日保藏在中国典型培养物保藏中心，地址: 武汉 武汉大学，保藏编号是：CCTCC NO：M 2013714) 适量，接种到PDA固体斜面培养基上在28℃培养箱中培养2至3天。
取斜面培养2至3天的草酸青霉 (Penicillium oxalicum) IBPT-6适量，接种到装有400mL培养液 [ 培养液组成(克/升)：甘露醇20.0，酵母膏3.0，麦芽糖20.0，味精10.0，葡萄糖10.0，KH2PO4 0.5，MgSO4 0.3，NaCl 15.0 定容 ] 的1000mL锥形瓶中，28℃静止培养30天后，获得菌丝体和发酵液。
2 浸膏的获得
用纱布将菌丝体和发酵液分离。菌丝体用丙酮溶液（含20%~30%水）连续超声破壁3次，过滤去除残渣，得到含丙酮和水的菌丝体粗提物。减压浓缩去除丙酮，得到粗提物的水溶液，再以体积比1:2加入乙酸乙酯萃取3次，得乙酸乙酯粗提液，减压浓缩至近干得菌丝体浸膏36.5 g。
3 化合物的分离精制
菌丝体浸膏通过100-200目硅胶拌样，以石油醚：二氯甲烷：甲醇为梯度洗脱液，进行减压硅胶色谱柱层析。经过简单的薄层色谱分析，合并，分离成组分A-E。组分D (5.9 g) (二氯甲烷：甲醇v/v=100:1的洗脱物)以二氯甲烷：甲醇为梯度洗脱剂，进行加压柱硅胶色谱层析，经过薄层色谱分析后合并得到五个亚组分D1-D5。组分D3（1.1 g）以甲醇：酸水（含0.1%三氟乙酸）为梯度洗脱剂进行反向硅胶色谱层析，分离得到四个亚组分D3-1~D3-4。D3的亚组分D3-2（168 mg）通过半制备液相色谱在流动相为50%乙腈（含0.1%三氟乙酸）条件下分离得到所述化合物（2.8 mg）。
化合物：常温下为黄色油状物，高分辨电喷雾质谱HRESI-MS在m/z：639.1712处给出分子离子峰[M+H]+（calcd for C32H31O14，639.1714），637.1567处给出分子离子峰[M–H]–（calcd for C32H29O14，637.1557），推断其分子式为C32H30O14。1H和13C-NMR数据见表1，主要的COSY、HMBC和NOE信号见图1。
表1 化合物的1H和13C-NMR数据（500MHz 1H and 125 MHz 13C，in DMSO d6）
a,b 信号可以互换
实施例2 体外抗肿瘤活性的测试
1 实验样品及实验方法
被测样品溶液的配制 测试样品为上述实施例1中分离精制的化合物纯品。精密称取适量样品，用甲醇配制成所需浓度的溶液，供测活性。
细胞系及细胞的继代培养采用肿瘤细胞系，肿瘤细胞用含10% FBS的DMEM 培养基，在37℃于通入5% CO2的培养箱中继代培养。
细胞增殖抑制活性测试方法
四氮唑盐（MTT）法：取对数生长期的肿瘤细胞，将细胞密度调至每毫升2×105个细胞，按每孔200微升接种于96孔细胞培养板中，于37℃通入5% CO2的培养箱中培养4小时。每孔加入2微升的样品液或空白溶液，培养24小时之后，每孔加入MTT液（MTT的每毫升5毫克生理盐水溶液）10微升，继续培养4小时，37℃、2000转/分钟离心8分钟，吸取上清。每孔加入DMSO各100微升，在微量振荡器上振荡15分钟，至结晶完全溶解之后，利用MD公司生产的SPECTRAMAX Plus型酶标仪测定每孔在570 nm处的吸光（OD）值。在同一块96孔板中样品的每个浓度均设置三孔，另设置三孔的空白对照和无细胞凋零孔（如果药物有颜色要做相应药物浓度无细胞凋零）。各孔OD值先做相应无细胞凋零，再取三孔平均OD值按IR (%) =(OD空白对照-OD样品)/OD空白对照×100% 计算每个浓度下细胞的增殖抑制率（IR%）。
2. 实验结果
细胞增殖抑制活性测试结果
在MTT法测试中，根据不同浓度的该化合物的肿瘤细胞增殖抑制率，应用SPSS16.0软件进行数据处理并计算半数抑制浓度IC50值。结果见表2。
表2 化合物对人癌细胞增殖的抑制活性(IC50，μM)
3. 结论
该化合物对多种人癌细胞具有较好的抗肿瘤活性。可作为制备癌细胞增殖抑制药物或抗肿瘤药物用于研究。
以上所述仅为本发明的较佳实施例，凡依本发明申请专利范围所做的均等变化与修饰，皆应属本发明的涵盖范围。
Claims (3)
1.化合物。
2.权利要求1所述的化合物在制备抑制人癌细胞增殖药物中的应用。
3.权利要求1所述的化合物在制备抗人癌药物中的应用。
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201910344814.XA CN110407793B (zh) | 2019-04-26 | 2019-04-26 | 源于草酸青霉的黑麦酮酸j及在抑制癌细胞增殖上的应用 |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201910344814.XA CN110407793B (zh) | 2019-04-26 | 2019-04-26 | 源于草酸青霉的黑麦酮酸j及在抑制癌细胞增殖上的应用 |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN110407793A true CN110407793A (zh) | 2019-11-05 |
| CN110407793B CN110407793B (zh) | 2022-08-12 |
Family
ID=68357696
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201910344814.XA Active CN110407793B (zh) | 2019-04-26 | 2019-04-26 | 源于草酸青霉的黑麦酮酸j及在抑制癌细胞增殖上的应用 |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN110407793B (zh) |
Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN110407792A (zh) * | 2019-04-26 | 2019-11-05 | 福州大学 | 源于草酸青霉的黑麦酮酸类化合物Secalonic acid J及制备方法 |
-
2019
- 2019-04-26 CN CN201910344814.XA patent/CN110407793B/zh active Active
Patent Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN110407792A (zh) * | 2019-04-26 | 2019-11-05 | 福州大学 | 源于草酸青霉的黑麦酮酸类化合物Secalonic acid J及制备方法 |
Also Published As
| Publication number | Publication date |
|---|---|
| CN110407793B (zh) | 2022-08-12 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN107298671B (zh) | 源于草酸青霉的黑麦酮酸h及制备抗人结肠癌药物的应用 | |
| CN107353274B (zh) | 源于草酸青霉的黑麦酮酸i及制备抗人食管癌药物的应用 | |
| CN107298672B (zh) | 源于草酸青霉的黑麦酮酸i在制备抗人结肠癌药物的应用 | |
| CN107485607B (zh) | 源于草酸青霉的黑麦酮酸h在制备抗人食管癌药物的应用 | |
| CN107298670B (zh) | 源于草酸青霉黑麦酮酸h制备抗人口腔表皮样癌药物应用 | |
| CN107298669B (zh) | 源于草酸青霉的黑麦酮酸i及抗人口腔表皮样癌药物应用 | |
| CN110407792A (zh) | 源于草酸青霉的黑麦酮酸类化合物Secalonic acid J及制备方法 | |
| CN109106702A (zh) | 源于草酸青霉的4-4’异构化黑麦酮酸d在结肠癌方面的应用 | |
| CN109134416B (zh) | 源于草酸青霉的黑麦酮酸h在制备人子宫颈癌药物的应用 | |
| CN109776477A (zh) | 源于草酸青霉的iso-Penicillixanthone A及抗长春新碱耐药应用 | |
| CN110407794A (zh) | 源于草酸青霉的黑麦酮酸k及在抑制癌细胞增殖上的应用 | |
| CN110407797A (zh) | 源于草酸青霉的黑麦酮酸类化合物Secalonic acid K及制备方法 | |
| CN109134417B (zh) | 源于草酸青霉的黑麦酮酸i及抗人子宫颈癌药物的应用 | |
| CN110407793A (zh) | 源于草酸青霉的黑麦酮酸j及在抑制癌细胞增殖上的应用 | |
| CN110407795A (zh) | 源于草酸青霉黑麦酮酸l及在抑制人癌细胞增殖上的应用 | |
| CN110403929A (zh) | 源于草酸青霉的黑麦酮酸m及在抑制人癌细胞增殖上的应用 | |
| CN110407798A (zh) | 源于草酸青霉的黑麦酮酸类化合物Secalonic acid M及制备方法 | |
| CN110407796A (zh) | 源于草酸青霉的黑麦酮酸类化合物Secalonic acid L及制备方法 | |
| CN106389418B (zh) | 源于桔绿木霉的青霉烯醇e1在肝癌方面的应用 | |
| CN106491596B (zh) | 源于桔绿木霉的青霉烯醇e1在肺癌方面的应用 | |
| CN105061443B (zh) | 源于桔青霉的penicitrinine A及其制备抗人肝癌药物的应用 | |
| CN105131006B (zh) | 源于桔青霉的penicitrinine A及在制备抗恶性黑色素瘤药物上的应用 | |
| CN109106703A (zh) | 源于草酸青霉的4-4’异构化黑麦酮酸d在鼻咽癌方面的应用 | |
| CN109106701A (zh) | 源于草酸青霉的4-4’异构化黑麦酮酸d在淋巴瘤方面的应用 | |
| CN106491597A (zh) | 源于桔绿木霉的青霉烯醇e1在食管癌方面的应用 |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| GR01 | Patent grant | ||
| GR01 | Patent grant | ||
| TR01 | Transfer of patent right | ||
| TR01 | Transfer of patent right |
Effective date of registration: 20240722 Address after: 350300 Nangang Avenue, Jiangyin Port City Economic Zone, Fuzhou City, Fujian Province, China Patentee after: Fujian Kangnong Pharmaceutical Technology Co.,Ltd. Country or region after: China Address before: No.2 Xueyuan Road, Fuzhou University Town, Shangjie Town, Minhou County, Fuzhou City, Fujian Province Patentee before: FUZHOU University Country or region before: China |