CN106432168A - 红树尖瓣海莲内生真菌来源的抗弧菌活性化合物及其制备方法 - Google Patents
红树尖瓣海莲内生真菌来源的抗弧菌活性化合物及其制备方法 Download PDFInfo
- Publication number
- CN106432168A CN106432168A CN201610829377.7A CN201610829377A CN106432168A CN 106432168 A CN106432168 A CN 106432168A CN 201610829377 A CN201610829377 A CN 201610829377A CN 106432168 A CN106432168 A CN 106432168A
- Authority
- CN
- China
- Prior art keywords
- compound
- vibrio
- pharmaceutically acceptable
- antibacterials
- preparation
- 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
- 238000002360 preparation method Methods 0.000 title claims abstract description 8
- 241000607598 Vibrio Species 0.000 title claims abstract description 7
- 150000001875 compounds Chemical class 0.000 title abstract description 20
- 241000233866 Fungi Species 0.000 title abstract description 4
- 241000039757 Bruguiera x rhynchopetala Species 0.000 title abstract 2
- 150000003839 salts Chemical class 0.000 claims abstract description 11
- 241000588724 Escherichia coli Species 0.000 claims abstract description 5
- 241000191963 Staphylococcus epidermidis Species 0.000 claims abstract description 5
- -1 isocoumarins compound Chemical class 0.000 claims abstract description 4
- OKKJLVBELUTLKV-UHFFFAOYSA-N Methanol Chemical compound OC OKKJLVBELUTLKV-UHFFFAOYSA-N 0.000 claims description 45
- 230000000844 anti-bacterial effect Effects 0.000 claims description 20
- 238000000855 fermentation Methods 0.000 claims description 18
- 230000004151 fermentation Effects 0.000 claims description 18
- 235000002639 sodium chloride Nutrition 0.000 claims description 18
- 239000013078 crystal Substances 0.000 claims description 16
- XEKOWRVHYACXOJ-UHFFFAOYSA-N Ethyl acetate Chemical compound CCOC(C)=O XEKOWRVHYACXOJ-UHFFFAOYSA-N 0.000 claims description 15
- 239000007788 liquid Substances 0.000 claims description 10
- 241000228153 Penicillium citrinum Species 0.000 claims description 9
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 claims description 9
- 229940088710 antibiotic agent Drugs 0.000 claims description 9
- 229940125904 compound 1 Drugs 0.000 claims description 9
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 claims description 8
- 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 8
- 239000001888 Peptone Substances 0.000 claims description 8
- 108010080698 Peptones Proteins 0.000 claims description 8
- 229940041514 candida albicans extract Drugs 0.000 claims description 8
- 239000008103 glucose Substances 0.000 claims description 8
- 239000001963 growth medium Substances 0.000 claims description 8
- 235000019319 peptone Nutrition 0.000 claims description 8
- 239000011780 sodium chloride Substances 0.000 claims description 8
- 239000012138 yeast extract Substances 0.000 claims description 8
- HAEQAUJYNHQVHV-UHFFFAOYSA-N 3-[4-(aminomethyl)-6-(trifluoromethyl)pyridin-2-yl]oxy-N-phenylbenzamide Chemical compound NCC1=CC(=NC(=C1)C(F)(F)F)OC=1C=C(C(=O)NC2=CC=CC=C2)C=CC=1 HAEQAUJYNHQVHV-UHFFFAOYSA-N 0.000 claims description 7
- 239000012071 phase Substances 0.000 claims description 7
- 241000894006 Bacteria Species 0.000 claims description 6
- HEDRZPFGACZZDS-UHFFFAOYSA-N Chloroform Chemical group ClC(Cl)Cl HEDRZPFGACZZDS-UHFFFAOYSA-N 0.000 claims description 6
- 238000010898 silica gel chromatography Methods 0.000 claims description 6
- SRCZQMGIVIYBBJ-UHFFFAOYSA-N ethoxyethane;ethyl acetate Chemical compound CCOCC.CCOC(C)=O SRCZQMGIVIYBBJ-UHFFFAOYSA-N 0.000 claims description 5
- 238000011534 incubation Methods 0.000 claims description 5
- 239000003208 petroleum Substances 0.000 claims description 5
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 5
- 229920001817 Agar Polymers 0.000 claims description 4
- 239000008272 agar Substances 0.000 claims description 4
- 238000000605 extraction Methods 0.000 claims description 4
- 238000004128 high performance liquid chromatography Methods 0.000 claims description 4
- 239000002609 medium Substances 0.000 claims description 4
- 238000000034 method Methods 0.000 claims description 4
- 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
- 241000193830 Bacillus <bacterium> Species 0.000 claims description 3
- 238000001641 gel filtration chromatography Methods 0.000 claims description 3
- 239000007791 liquid phase Substances 0.000 claims description 3
- 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 claims description 3
- 230000006837 decompression Effects 0.000 claims description 2
- 239000003937 drug carrier Substances 0.000 claims description 2
- 238000010828 elution Methods 0.000 claims description 2
- 238000010899 nucleation Methods 0.000 claims description 2
- 230000006911 nucleation Effects 0.000 claims description 2
- 230000002829 reductive effect Effects 0.000 claims description 2
- 230000002269 spontaneous effect Effects 0.000 claims description 2
- 239000000126 substance Substances 0.000 claims description 2
- AOSZTAHDEDLTLQ-AZKQZHLXSA-N (1S,2S,4R,8S,9S,11S,12R,13S,19S)-6-[(3-chlorophenyl)methyl]-12,19-difluoro-11-hydroxy-8-(2-hydroxyacetyl)-9,13-dimethyl-6-azapentacyclo[10.8.0.02,9.04,8.013,18]icosa-14,17-dien-16-one Chemical compound C([C@@H]1C[C@H]2[C@H]3[C@]([C@]4(C=CC(=O)C=C4[C@@H](F)C3)C)(F)[C@@H](O)C[C@@]2([C@@]1(C1)C(=O)CO)C)N1CC1=CC=CC(Cl)=C1 AOSZTAHDEDLTLQ-AZKQZHLXSA-N 0.000 claims 1
- UNILWMWFPHPYOR-KXEYIPSPSA-M 1-[6-[2-[3-[3-[3-[2-[2-[3-[[2-[2-[[(2r)-1-[[2-[[(2r)-1-[3-[2-[2-[3-[[2-(2-amino-2-oxoethoxy)acetyl]amino]propoxy]ethoxy]ethoxy]propylamino]-3-hydroxy-1-oxopropan-2-yl]amino]-2-oxoethyl]amino]-3-[(2r)-2,3-di(hexadecanoyloxy)propyl]sulfanyl-1-oxopropan-2-yl Chemical compound O=C1C(SCCC(=O)NCCCOCCOCCOCCCNC(=O)COCC(=O)N[C@@H](CSC[C@@H](COC(=O)CCCCCCCCCCCCCCC)OC(=O)CCCCCCCCCCCCCCC)C(=O)NCC(=O)N[C@H](CO)C(=O)NCCCOCCOCCOCCCNC(=O)COCC(N)=O)CC(=O)N1CCNC(=O)CCCCCN\1C2=CC=C(S([O-])(=O)=O)C=C2CC/1=C/C=C/C=C/C1=[N+](CC)C2=CC=C(S([O-])(=O)=O)C=C2C1 UNILWMWFPHPYOR-KXEYIPSPSA-M 0.000 claims 1
- FFRBMBIXVSCUFS-UHFFFAOYSA-N 2,4-dinitro-1-naphthol Chemical compound C1=CC=C2C(O)=C([N+]([O-])=O)C=C([N+]([O-])=O)C2=C1 FFRBMBIXVSCUFS-UHFFFAOYSA-N 0.000 claims 1
- 241001478240 Coccus Species 0.000 claims 1
- 229940126657 Compound 17 Drugs 0.000 claims 1
- 235000009754 Vitis X bourquina Nutrition 0.000 claims 1
- 235000012333 Vitis X labruscana Nutrition 0.000 claims 1
- 240000006365 Vitis vinifera Species 0.000 claims 1
- 235000014787 Vitis vinifera Nutrition 0.000 claims 1
- QTBSBXVTEAMEQO-UHFFFAOYSA-N acetic acid Substances CC(O)=O QTBSBXVTEAMEQO-UHFFFAOYSA-N 0.000 claims 1
- 150000002148 esters Chemical class 0.000 claims 1
- 239000004615 ingredient Substances 0.000 claims 1
- 241000191967 Staphylococcus aureus Species 0.000 abstract description 3
- 230000002401 inhibitory effect Effects 0.000 abstract description 2
- 241000193755 Bacillus cereus Species 0.000 abstract 1
- 229940124350 antibacterial drug Drugs 0.000 abstract 1
- IQZZFVDIZRWADY-UHFFFAOYSA-N isocumarine Natural products C1=CC=C2C(=O)OC=CC2=C1 IQZZFVDIZRWADY-UHFFFAOYSA-N 0.000 abstract 1
- 229930014626 natural product Natural products 0.000 abstract 1
- 240000002044 Rhizophora apiculata Species 0.000 description 8
- 241000409653 Bruguiera sexangula Species 0.000 description 7
- 229930000044 secondary metabolite Natural products 0.000 description 4
- 239000011734 sodium Substances 0.000 description 4
- 230000001580 bacterial effect Effects 0.000 description 3
- 238000002447 crystallographic data Methods 0.000 description 3
- 238000002114 high-resolution electrospray ionisation mass spectrometry Methods 0.000 description 3
- 239000000047 product Substances 0.000 description 3
- 206010059866 Drug resistance Diseases 0.000 description 2
- 230000005260 alpha ray Effects 0.000 description 2
- 229940125782 compound 2 Drugs 0.000 description 2
- 229940126214 compound 3 Drugs 0.000 description 2
- 239000003814 drug Substances 0.000 description 2
- 230000002538 fungal effect Effects 0.000 description 2
- 244000005700 microbiome Species 0.000 description 2
- 239000000203 mixture Substances 0.000 description 2
- 239000002798 polar solvent Substances 0.000 description 2
- 230000003068 static effect Effects 0.000 description 2
- 238000001644 13C nuclear magnetic resonance spectroscopy Methods 0.000 description 1
- 238000005160 1H NMR spectroscopy Methods 0.000 description 1
- 240000001817 Cereus hexagonus Species 0.000 description 1
- 241000196324 Embryophyta Species 0.000 description 1
- 241000192125 Firmicutes Species 0.000 description 1
- 238000005481 NMR spectroscopy Methods 0.000 description 1
- 241000607594 Vibrio alginolyticus Species 0.000 description 1
- 150000001447 alkali salts Chemical class 0.000 description 1
- 239000003242 anti bacterial agent Substances 0.000 description 1
- 244000052616 bacterial pathogen Species 0.000 description 1
- 239000002585 base Substances 0.000 description 1
- 230000003115 biocidal effect Effects 0.000 description 1
- 238000012822 chemical development Methods 0.000 description 1
- 229940125898 compound 5 Drugs 0.000 description 1
- 238000002425 crystallisation Methods 0.000 description 1
- 230000008025 crystallization Effects 0.000 description 1
- 238000011161 development Methods 0.000 description 1
- 229940079593 drug Drugs 0.000 description 1
- 238000012912 drug discovery process Methods 0.000 description 1
- 230000000694 effects Effects 0.000 description 1
- 238000005516 engineering process Methods 0.000 description 1
- 230000007613 environmental effect Effects 0.000 description 1
- 238000003810 ethyl acetate extraction Methods 0.000 description 1
- 239000000284 extract Substances 0.000 description 1
- 239000000706 filtrate Substances 0.000 description 1
- 238000001914 filtration Methods 0.000 description 1
- 125000002791 glucosyl group Chemical group C1([C@H](O)[C@@H](O)[C@H](O)[C@H](O1)CO)* 0.000 description 1
- 238000005286 illumination Methods 0.000 description 1
- 150000002611 lead compounds Chemical class 0.000 description 1
- 230000000670 limiting effect Effects 0.000 description 1
- 239000000463 material Substances 0.000 description 1
- 150000007522 mineralic acids Chemical class 0.000 description 1
- 150000007524 organic acids Chemical class 0.000 description 1
- AICOOMRHRUFYCM-ZRRPKQBOSA-N oxazine, 1 Chemical compound C([C@@H]1[C@H](C(C[C@]2(C)[C@@H]([C@H](C)N(C)C)[C@H](O)C[C@]21C)=O)CC1=CC2)C[C@H]1[C@@]1(C)[C@H]2N=C(C(C)C)OC1 AICOOMRHRUFYCM-ZRRPKQBOSA-N 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
- C07D311/76—Benzo[c]pyrans
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07D—HETEROCYCLIC COMPOUNDS
- C07D493/00—Heterocyclic compounds containing oxygen atoms as the only ring hetero atoms in the condensed system
- C07D493/12—Heterocyclic compounds containing oxygen atoms as the only ring hetero atoms in the condensed system in which the condensed system contains three hetero rings
- C07D493/20—Spiro-condensed systems
-
- 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
-
- 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
- C12P17/18—Preparation of heterocyclic carbon compounds with only O, N, S, Se or Te as ring hetero atoms containing at least two hetero rings condensed among themselves or condensed with a common carbocyclic ring system, e.g. rifamycin
- C12P17/181—Heterocyclic compounds containing oxygen atoms as the only ring heteroatoms in the condensed system, e.g. Salinomycin, Septamycin
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07B—GENERAL METHODS OF ORGANIC CHEMISTRY; APPARATUS THEREFOR
- C07B2200/00—Indexing scheme relating to specific properties of organic compounds
- C07B2200/13—Crystalline forms, e.g. polymorphs
-
- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
- Y02A—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE
- Y02A50/00—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE in human health protection, e.g. against extreme weather
- Y02A50/30—Against vector-borne diseases, e.g. mosquito-borne, fly-borne, tick-borne or waterborne diseases whose impact is exacerbated by climate change
Abstract
本发明属于海洋天然产物领域,具体涉及红树尖瓣海莲内生真菌来源的抗弧菌活性化合物及其制备方法。本发明提供一种异香豆素类化合物1‑3或其药学上可接受的盐,化合物1‑3具有如下结构:
Description
技术领域
本发明属于海洋天然产物领域,具体涉及一种由红树尖瓣海莲内生真菌产生的抗弧菌活性化合物及其制备方法。
背景技术
在过去的半个多世纪里,由于抗生素的广泛使用,特别是不合理的临床误用,造成耐药性致病菌种类的不断增加,对人类生命健康带来极大威胁。为了减少细菌耐药性的产生,急需开发新的高敏抗菌药物,以提高临床用药的有效性。
海洋高盐、高压、低光照等条件使海洋微生物能够产生大量结构新颖、活性独特的次级代谢产物,其中许多化合物具有抗菌活性,为寻找潜在的抗菌药物先导化合物提供了重要来源。
近年来,随着海洋天然药物化学的发展,越来越多的抗菌活性化合物从海洋微生物中不断分离获得,为寻找新型抗菌剂提供了物质基础。采用人工培养发酵的方法,从海洋微生物中获得有重要抗菌活性的次级代谢产物,具有环境友好、可持续发展等特点,能有效解决药物开发过程中的药源等关键性问题,因此具有独特优势。
发明内容
本发明提供一种真菌Penicillium citrinum HL-5126分离自红树尖瓣海莲,红树尖瓣海莲是第一发明人于2012年采自中国海南东寨港红树林自然保护区(NaturalProduct Research,2016,30(7),821-825)。
本发明提供一类由上述红树尖瓣海莲内生真菌Penicillium citrinum HL-5126产生的次级代谢产物或其药学上可接受的盐,其特征在于所述次级代谢产物具有如下结构:
本发明提供一种同时制备化合物1-7或其药学上可接受的盐的制备方法,其特征在于包括如下步骤:
(1)先在菌种培养基中对真菌Penicillium citrinum HL-5126进行菌种培养;
(2)再在发酵培养基中对步骤(1)中的真菌Penicillium citrinum HL-5126进行发酵培养;
(3)将步骤(2)得到的发酵物中的发酵液和菌体分离,发酵液用等体积的乙酸乙酯萃取3~5次,合并萃取液后减压浓缩得到发酵液浸膏;
(4)步骤(3)得到的发酵液浸膏经过减压硅胶柱层析,采用石油醚-乙酸乙酯按100:0至0:100梯度洗脱,将其按照极性大小分成5个组分Fr.1~Fr.5;将Fr.3组分以石油醚-乙酸乙酯10∶2为洗脱剂经正相硅胶柱层析后,再经Sephadex LH-20凝胶柱层析,洗脱剂为CHCl3∶MeOH=1∶1,最后经高效液相色谱HPLC制备,所用色谱柱为Agilent C18,9.4×250mm,7μm,流速为2mL/min,流动相为MeOH∶H2O=85∶15,最终得到化合物1-7。
其中所述洗脱剂或流动相的比例均为体积比;所述菌种培养基中含有葡萄糖0.1%–5.0%、酵母膏0.01%–2%、蛋白胨0.01%–2%、琼脂0.1%–3.0%、粗海盐0.05%–5%、适量的水,培养温度为20–25℃,培养时间为5-10天;所述发酵培养基中含有葡萄糖0.1%–5.0%、酵母膏0.01%–2%、蛋白胨0.01%–2%、粗海盐0.05%–5%,培养温度为20–25℃,培养时间为25–30天;上述百分比均为重量百分比。
本发明提供化合物1的晶体结构,其特征在于其X-射线单晶衍射数据为: 斜方晶系,空间群P212121,晶胞参数为 α=90°,β=90°,γ=90°,Z=4,Dx=1.318mg/mm3,F(000)=640,μ(Cu Kα)=0.848mm-1,2681个可观测点[I>2σ(I)],可观测点精修最终偏离因子R=0.083,wR=0.0707,Flack常数为0.0(3)。
本发明提供化合物1晶体的制备方法,其特征在于将化合物1溶于极性溶剂中,自然结晶得到,所述极性溶剂为甲醇、乙醇或乙酸乙酯。
本发明提供一种抗菌剂,其特征在于包含化合物1-7、其晶体或其药学上可接受的盐作为活性成分。
本发明提供的上述抗菌剂,还可以包括其他抗菌药物;也可以包括药学上可接受的载体。
本发明提供化合物1-7、其晶体或其药学上可接受的盐在制备抗菌药物方面的用途。所述抗菌药物用于抑制表皮葡萄球菌、金黄色葡萄球菌、大肠杆菌、蜡状芽孢杆菌或弧菌。
本发明所述晶体结构以双子超衍射仪(Xcalibur,Atlas,Gemini ultradiffractometer)采用Cu Kα射线在120.01(10)K下扫描收集衍射数据。
本发明中术语“药学上可接受的盐”是指非毒性的无机或有机酸和/或碱的加成盐,可参见“Salt selection for basic drugs”,Int.J.Pharm.(1986),33,201–217。
附图说明
说明书附图1为化合物1的XRD图。
具体实施方式
为了便于对本发明的进一步理解,下面提供的实施例对其做了更详细的说明。但是这些实施例仅供更好的理解发明而并非用来限定本发明的范围或实施原则,本发明的实施方式不限于以下内容。
实施例1
(1)红树尖瓣海莲内生真菌Penicillium citrinum HL-5126的菌种培养
菌种培养所用的培养基含有葡萄糖5.0%(重量百分比,下同)、酵母膏2%、蛋白胨2%、琼脂3%、粗海盐5.0%,其余为水;使用时制成试管斜面,真菌菌株在25℃下培养5天。
(2)红树尖瓣海莲内生真菌Penicillium citrinum HL-5126的发酵
发酵培养所用的培养基含有葡萄糖5.0%(重量百分比,下同)、酵母膏2%、蛋白胨2%、氯化钠5%,其余为水;真菌菌株于25℃培养30天。
(3)化合物1-7的提取分离
取20L步骤(2)得到的发酵液过滤,除去菌体,将滤液浓缩后,用等体积的乙酸乙酯萃取3次;萃取液浓缩后经过减压硅胶柱层析,采用石油醚-乙酸乙酯按100:0至0∶100梯度洗脱,将其按照极性大小分成5个组分Fr.1~Fr.5;将Fr.3组分以石油醚-乙酸乙酯10∶2为洗脱剂经正相硅胶柱层析后,再经Sephadex LH-20凝胶柱层析,洗脱剂为CHCl3∶MeOH=1∶1,最后经高效液相色谱HPLC制备,色谱柱为Agilent C18,9.4×250mm,7μm,流速为2mL/min,流动相为MeOH∶H2O=85∶15,最终得到化合物1(5.0mg)、化合物2(5.0mg)、化合物3(6.2mg)、化合物4(6.0mg)、化合物5(8.0mg)、化合物6(10.0mg)、化合物7(7.0mg)。
经结构确证,化合物1-3为新化合物,化合物4-7表征数据与已知报道的数据一致,在本发明中不在赘述。
化合物1-3的NMR数据(400MHz 1H NMR,100MHz 13C NMR)
化合物1:无色晶体,[α]24 D=–29.0(c=0.25,MeOH).mp 118.8–120.4℃;λmax(logε)220(3.02),246(2.50),312(1.62)nm;CD(c 2×10-4mol/L,MeOH)λmax(Δε),259(-23.0),280(-2.5),330(-13.6);IR(KBr)νmax 3518,1712,1602,1272,1250,1230,1112,1082,1064,798cm–1;HRESIMS m/z 281.1384[M+H]+(calcd for C15H21O5,281.1383).
化合物2:无色晶体,[α]24 D=–24.6(c=0.25,MeOH).mp 136.9–137.6℃.λmax(logε)220(3.02),246(2.50),312(1.62)nm;CD(0.52Mm,MeOH)λmax(Δε)239(+2.00),257(-8.60),290(-1.52),329(-3.10);IR(KBr)νmax 3518,1712,1602,1272,1250,1230,1112,1082,1064,798cm–1;HRESIMS m/z 579.2200[2M+Na]+(calcd for C30H36O10Na,579.2200).
化合物3:无色晶体,[α]24 D=–12.0(c=0.25,MeOH).λmax(logε)220(3.02),246(2.50),312(1.62)nm;CD(0.52Mm,MeOH)λmax(Δε)240(+9.60),260(-2.65),328(-0.16);IR(KBr)νmax 3518,1712,1602,1272,1250,1230,1112,1082,1064,798cm–1;HRESIMS m/z273.0733[M+Na]+(calcd for C30H36O10Na,273.0740).
实施例2
将实施例1中菌种培养基中成分调整为葡萄糖0.1%、酵母膏0.01%、蛋白胨0.01%、琼脂0.1%、粗海盐0.05%、适量的水,培养温度为20℃,培养时间为10天;所述发酵培养基中成分调整为含有葡萄糖0.1%、酵母膏0.01%、蛋白胨 0.01%、粗海盐0.05%,培养温度为25℃,培养时间为25天;上述百分比均为重量百分比。仍能以类似的收率得到化合物1-7。
实施例3
取2.0mg化合物1溶于2mL甲醇中,静止自然结晶,4天后得到无色晶体,其晶体结构以双子超衍射仪(Xcalibur,Atlas,Gemini ultra diffractometer)采用Cu Kα射线在120.01(10)K下扫描收集衍射数据。
化合物1晶体:斜方晶系,空间群P212121,晶胞参数为 α=90°,β=90°,γ=90°,Z=4,Dx=1.318mg/mm3,F(000)=640,μ(Cu Kα)=0.848mm-1,2681个可观测点[I>2σ(I)],可观测点精修最终偏离因子R=0.083,wR=0.0707,Flack常数为0.0(3)。
此外,将化合物1溶于乙醇或乙酸乙酯中,静止3-7天均能得到上述晶体。
实施例4本发明的化合物的抗菌活性的测定
本发明的化合物按照文献方法(Pierce C.G.;Uppuluri P.;Teistan A.R.;Wormley Jr.F.L.;Mowat E.;Ramage G.;Lopez-ribot J.L.Nat.Protoc.2008,3,1494-1500),测试对4株革兰氏阳性菌:蜡状芽孢杆菌(B.cereus)、金黄色葡萄球菌(S.aureus)、表皮葡萄球菌(S.epidermidis)、大肠杆菌(Escherichia coli)、弧菌(V.alginolyticus)的抗菌活性。如表1所示。
表1本发明的化合物对细菌的抑制活性
Claims (10)
1.一种异香豆素类化合物1-3或其药学上可接受的盐,其特征在于所述异香豆素类化合物1-3具有如下结构:
2.一种利用真菌Penicillium citrinum HL-5126同时制备化合物1-7的方法,其特征在于包括如下步骤:
(1)先在菌种培养基中对真菌Penicillium citrinum HL-5126进行菌种培养;
(2)再在发酵培养基中对步骤(1)中的真菌Penicillium citrinum HL-5126进行发酵培养;
(3)将步骤(2)得到的发酵物中的发酵液和菌体分离,发酵液用等体积的乙酸乙酯萃取3~5次,合并萃取液后减压浓缩得到发酵液浸膏;
(4)步骤(3)得到的发酵液浸膏经过减压硅胶柱层析,采用石油醚-乙酸乙酯按100:0至0∶100梯度洗脱,将其按照极性大小分成5个组分Fr.1~Fr.5;将Fr.3组分以石油醚-乙酸乙酯10∶2为洗脱剂经正相硅胶柱层析后,再经Sephadex LH-20凝胶柱层析,洗脱剂为CHCl3∶MeOH=1∶1,最后经高效液相色谱HPLC制备,得到化合物1-7;所述HPLC制备的色谱条件为:色谱柱为Agilent C18,9.4×250mm,7μm,流速为2mL/min,流动相为MeOH∶H2O=85∶15;化合物1-7具有如下结构:
3.权利要求2所述的制备方法,其特征在于所述菌种培养基中含有葡萄糖0.1%–5.0%、酵母膏0.01%–2%、蛋白胨0.01%–2%、琼脂0.1%–3.0%、粗海盐0.05%–5%、适量的水,培养温度为20–25℃,培养时间为5-10天;所述发酵培养基中含有葡萄糖0.1%–5.0%、酵母膏0.01%–2%、蛋白胨0.01%–2%、粗海盐0.05%–5%,培养温度为20–25℃,培养时间为25–30天;上述百分比均为重量百分比。
4.一种权利要求1所述的化合物1的晶体的制备方法,其特征在于包括如下步骤:
(1)按照权利要求2-3任一项所述的方法制备得到化合物1;
(2)将步骤(1)得到的化合物1溶于适量的甲醇、乙醇或乙酸乙酯中,自然结晶4-7天得到无色晶体;
所述无色晶体属于斜方晶系,空间群P212121,晶胞参数为 α=90°,β=90°,γ=90°,Z=4,Dx=1.318mg/mm3,F(000)=640,μ(Cu Kα)=0.848mm-1,2681个可观测点[I>2σ(I)],可观测点精修最终偏离因子R=0.083,wR=0.0707,Flack常数为0.0(3)。
5.一种抗菌剂,其特征在于以权利要求1所述的化合物1–3或其药学上可接受的盐作为有效成分。
6.权利要求5所述的抗菌剂,其特征在于还包含其他抗菌药物。
7.权利要求5-6任一项所述的抗菌剂,其特征在于还包括药学上可接受的载体。
8.权利要求2所述的化合物1–7或其药学上可接受的盐在制备抗菌药物中的应用。
9.权利要求8所述的应用,其特征在于所述抗菌药物用于抑制表皮葡萄球菌、金黄色葡萄球菌、大肠杆菌、蜡状芽孢杆菌或弧菌。
10.权利要求9所述的应用,其特征在于所述抗菌药物用于抑制弧菌。
Priority Applications (1)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
CN201610829377.7A CN106432168B (zh) | 2016-09-19 | 2016-09-19 | 红树尖瓣海莲内生真菌来源的抗弧菌活性化合物及其制备方法 |
Applications Claiming Priority (1)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
CN201610829377.7A CN106432168B (zh) | 2016-09-19 | 2016-09-19 | 红树尖瓣海莲内生真菌来源的抗弧菌活性化合物及其制备方法 |
Publications (2)
Publication Number | Publication Date |
---|---|
CN106432168A true CN106432168A (zh) | 2017-02-22 |
CN106432168B CN106432168B (zh) | 2019-01-11 |
Family
ID=58168558
Family Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
CN201610829377.7A Active CN106432168B (zh) | 2016-09-19 | 2016-09-19 | 红树尖瓣海莲内生真菌来源的抗弧菌活性化合物及其制备方法 |
Country Status (1)
Country | Link |
---|---|
CN (1) | CN106432168B (zh) |
Cited By (4)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN107827805A (zh) * | 2017-06-05 | 2018-03-23 | 海南师范大学 | 一种红树植物木果楝真菌来源的吲哚二萜类化合物及其制备方法与应用 |
CN108546651A (zh) * | 2018-04-24 | 2018-09-18 | 广东立威化工有限公司 | 红树植物秋茄内生真菌2cpe-1及其发酵液与应用 |
CN109553600A (zh) * | 2018-12-04 | 2019-04-02 | 海南师范大学 | 一种红树内生真菌中异香豆素类化合物及其制备方法与应用 |
CN115490661A (zh) * | 2022-08-09 | 2022-12-20 | 海南师范大学 | 红树来源真菌中抗氧化活性化合物及其制备方法 |
Citations (1)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN105481817A (zh) * | 2015-11-17 | 2016-04-13 | 云南民族大学 | 一种异香豆素类化合物及其制备方法和应用 |
-
2016
- 2016-09-19 CN CN201610829377.7A patent/CN106432168B/zh active Active
Patent Citations (1)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN105481817A (zh) * | 2015-11-17 | 2016-04-13 | 云南民族大学 | 一种异香豆素类化合物及其制备方法和应用 |
Non-Patent Citations (3)
Title |
---|
JHILLU SINGH YADAV ET AL.,: "First enantioselective total synthesis of penicimarin B,aspergillumarins A and B", 《TETRAHEDRON LETTERS》 * |
JIRAPORN ARUNPANICHLERT ET AL.,: "Meroterpenoid,isocoumarin,and phenol derivatives from the seagrass-derived fungus Pestalotiopsis sp.PSU-ES194", 《TETRAHEDRON》 * |
黄国雷 等,: "从尖瓣海莲内生真菌中分离得到的异香豆素类化合物", 《中国化学会第30届学术年会摘要集-第九分会:有机化学》 * |
Cited By (6)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN107827805A (zh) * | 2017-06-05 | 2018-03-23 | 海南师范大学 | 一种红树植物木果楝真菌来源的吲哚二萜类化合物及其制备方法与应用 |
CN108546651A (zh) * | 2018-04-24 | 2018-09-18 | 广东立威化工有限公司 | 红树植物秋茄内生真菌2cpe-1及其发酵液与应用 |
CN108546651B (zh) * | 2018-04-24 | 2020-11-10 | 广东立威化工有限公司 | 红树植物秋茄内生真菌2cpe-1及其发酵液与应用 |
CN109553600A (zh) * | 2018-12-04 | 2019-04-02 | 海南师范大学 | 一种红树内生真菌中异香豆素类化合物及其制备方法与应用 |
CN115490661A (zh) * | 2022-08-09 | 2022-12-20 | 海南师范大学 | 红树来源真菌中抗氧化活性化合物及其制备方法 |
CN115490661B (zh) * | 2022-08-09 | 2023-09-08 | 海南师范大学 | 红树来源真菌中抗氧化活性化合物及其制备方法 |
Also Published As
Publication number | Publication date |
---|---|
CN106432168B (zh) | 2019-01-11 |
Similar Documents
Publication | Publication Date | Title |
---|---|---|
CN106432168A (zh) | 红树尖瓣海莲内生真菌来源的抗弧菌活性化合物及其制备方法 | |
CN107827805B (zh) | 一种红树植物木果楝真菌来源的吲哚二萜类化合物及其制备方法与应用 | |
CN110527629B (zh) | 一种海洋真菌来源的布雷菲德菌素a、制备方法及其在抗农业致病菌方面的应用 | |
CN115490661B (zh) | 红树来源真菌中抗氧化活性化合物及其制备方法 | |
CN115595343A (zh) | 樱桃番茄根际土壤微生物中活性次级代谢产物及其制备与应用 | |
CN106432169B (zh) | 一种抗弧菌活性的异香豆素类化合物及其晶体 | |
CN107828663B (zh) | 一种吲哚二萜类化合物晶体及其作为抗肿瘤药物的应用 | |
CN110330544B (zh) | 一种4,4,1-双环甾类化合物及其制备方法和用途 | |
CN110357788B (zh) | 一种聚酮类化合物及其制备方法和用途 | |
CN102079692B (zh) | 一种三联苯化合物及其制备方法和作为α-葡萄糖苷酶抑制剂的应用 | |
CN110407792B (zh) | 源于草酸青霉的黑麦酮酸类化合物Secalonic acid J及制备方法 | |
CN115724816B (zh) | 红树来源真菌中色原酮晶型及其制备与应用 | |
CN103880826A (zh) | 一种异苯并呋喃酮化合物及其制备方法和应用 | |
CN103555806A (zh) | 一种高效利用亚麻刺盘孢霉合成7α-羟基-雄甾烯酮的方法 | |
CN108727169B (zh) | 一种海洋真菌来源的联苯醚化合物的制备方法与作为抗菌剂的应用 | |
JPS61216692A (ja) | Cl‐1577‐b↓4化合物およびその製法 | |
CN114621092A (zh) | 一种红树植物来源真菌中酚类化合物及其制备方法 | |
CN107973803B (zh) | 一种七元内酯并呋喃衍生物及其制备方法和应用 | |
CN107226800A (zh) | 一种xanthone类化合物及其单晶的制备方法与作为抗海分枝杆菌药物的应用 | |
CN107226801A (zh) | 一种xanthone类化合物及其单晶的制备方法与作为α-葡萄糖苷酶抑制剂的应用 | |
CN108794502B (zh) | 一种单端孢霉烯类化合物及其制备方法和用途 | |
CN103214449A (zh) | 一种氯代二苯甲酮化合物及其制备方法与应用 | |
CN110407797B (zh) | 源于草酸青霉的黑麦酮酸类化合物Secalonic acid K及制备方法 | |
CN104230874B (zh) | 一种异香豆素类化合物的制备方法与作为抗菌剂的应用 | |
CN108342325B (zh) | 一种蝉虫草来源的蒽醌类化合物及其制备方法和应用 |
Legal Events
Date | Code | Title | Description |
---|---|---|---|
C06 | Publication | ||
PB01 | Publication | ||
C10 | Entry into substantive examination | ||
SE01 | Entry into force of request for substantive examination | ||
GR01 | Patent grant | ||
GR01 | Patent grant |