CN113444644B - Application of naphtho-gamma-pyrone compounds in preparation of anti-helicobacter pylori medicines or prevention health-care products - Google Patents
Application of naphtho-gamma-pyrone compounds in preparation of anti-helicobacter pylori medicines or prevention health-care products Download PDFInfo
- Publication number
- CN113444644B CN113444644B CN202010223692.1A CN202010223692A CN113444644B CN 113444644 B CN113444644 B CN 113444644B CN 202010223692 A CN202010223692 A CN 202010223692A CN 113444644 B CN113444644 B CN 113444644B
- Authority
- CN
- China
- Prior art keywords
- naphtho
- gamma
- pyrone
- formula
- follows
- 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.)
- Active
Links
- 238000002360 preparation method Methods 0.000 title claims description 22
- 239000003814 drug Substances 0.000 title abstract description 30
- 229940037467 helicobacter pylori Drugs 0.000 title abstract description 28
- 229940079593 drug Drugs 0.000 title abstract description 25
- 230000002265 prevention Effects 0.000 title abstract description 8
- 238000000855 fermentation Methods 0.000 claims abstract description 32
- 230000004151 fermentation Effects 0.000 claims abstract description 32
- 239000000284 extract Substances 0.000 claims abstract description 22
- 241000233866 Fungi Species 0.000 claims abstract description 19
- 238000004321 preservation Methods 0.000 claims abstract description 11
- 241000228257 Aspergillus sp. Species 0.000 claims abstract description 10
- 238000000034 method Methods 0.000 claims abstract description 10
- CVQUWLDCFXOXEN-UHFFFAOYSA-N Pyran-4-one Chemical class O=C1C=COC=C1 CVQUWLDCFXOXEN-UHFFFAOYSA-N 0.000 claims abstract description 9
- OKKJLVBELUTLKV-UHFFFAOYSA-N Methanol Chemical compound OC OKKJLVBELUTLKV-UHFFFAOYSA-N 0.000 claims description 25
- XEKOWRVHYACXOJ-UHFFFAOYSA-N Ethyl acetate Chemical compound CCOC(C)=O XEKOWRVHYACXOJ-UHFFFAOYSA-N 0.000 claims description 23
- 240000008042 Zea mays Species 0.000 claims description 13
- 235000005824 Zea mays ssp. parviglumis Nutrition 0.000 claims description 13
- 235000002017 Zea mays subsp mays Nutrition 0.000 claims description 13
- 235000005822 corn Nutrition 0.000 claims description 13
- 241000228212 Aspergillus Species 0.000 claims description 12
- 229940041514 candida albicans extract Drugs 0.000 claims description 11
- 239000012138 yeast extract Substances 0.000 claims description 11
- CSNNHWWHGAXBCP-UHFFFAOYSA-L Magnesium sulfate Chemical compound [Mg+2].[O-][S+2]([O-])([O-])[O-] CSNNHWWHGAXBCP-UHFFFAOYSA-L 0.000 claims description 10
- -1 gamma-pyrone compound Chemical class 0.000 claims description 10
- CSCPPACGZOOCGX-UHFFFAOYSA-N Acetone Chemical compound CC(C)=O CSCPPACGZOOCGX-UHFFFAOYSA-N 0.000 claims description 9
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 claims description 8
- 235000002639 sodium chloride Nutrition 0.000 claims description 8
- 239000007836 KH2PO4 Substances 0.000 claims description 7
- 229910000402 monopotassium phosphate Inorganic materials 0.000 claims description 7
- 239000013535 sea water Substances 0.000 claims description 7
- 229910052943 magnesium sulfate Inorganic materials 0.000 claims description 6
- 239000000843 powder Substances 0.000 claims description 6
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 claims description 5
- 239000011780 sodium chloride Substances 0.000 claims description 5
- 230000003068 static effect Effects 0.000 claims description 5
- 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 4
- NGPGDYLVALNKEG-UHFFFAOYSA-N azanium;azane;2,3,4-trihydroxy-4-oxobutanoate Chemical compound [NH4+].[NH4+].[O-]C(=O)C(O)C(O)C([O-])=O NGPGDYLVALNKEG-UHFFFAOYSA-N 0.000 claims description 4
- 239000008103 glucose Substances 0.000 claims description 4
- 238000002386 leaching Methods 0.000 claims description 4
- 239000003960 organic solvent Substances 0.000 claims description 4
- 239000001888 Peptone Substances 0.000 claims description 3
- 108010080698 Peptones Proteins 0.000 claims description 3
- 235000013339 cereals Nutrition 0.000 claims description 3
- 235000019319 peptone Nutrition 0.000 claims description 3
- GNSKLFRGEWLPPA-UHFFFAOYSA-M potassium dihydrogen phosphate Chemical compound [K+].OP(O)([O-])=O GNSKLFRGEWLPPA-UHFFFAOYSA-M 0.000 claims description 3
- 244000061456 Solanum tuberosum Species 0.000 claims description 2
- 235000002595 Solanum tuberosum Nutrition 0.000 claims description 2
- 229920002472 Starch Polymers 0.000 claims description 2
- 229930006000 Sucrose Natural products 0.000 claims description 2
- CZMRCDWAGMRECN-UGDNZRGBSA-N Sucrose Chemical compound O[C@H]1[C@H](O)[C@@H](CO)O[C@@]1(CO)O[C@@H]1[C@H](O)[C@@H](O)[C@H](O)[C@@H](CO)O1 CZMRCDWAGMRECN-UGDNZRGBSA-N 0.000 claims description 2
- 230000003213 activating effect Effects 0.000 claims description 2
- 229910052564 epsomite Inorganic materials 0.000 claims description 2
- 235000013312 flour Nutrition 0.000 claims description 2
- 229910000359 iron(II) sulfate Inorganic materials 0.000 claims description 2
- 239000002245 particle Substances 0.000 claims description 2
- 235000012015 potatoes Nutrition 0.000 claims description 2
- VWDWKYIASSYTQR-UHFFFAOYSA-N sodium nitrate Inorganic materials [Na+].[O-][N+]([O-])=O VWDWKYIASSYTQR-UHFFFAOYSA-N 0.000 claims description 2
- 239000008107 starch Substances 0.000 claims description 2
- 235000019698 starch Nutrition 0.000 claims description 2
- 229960004793 sucrose Drugs 0.000 claims description 2
- 238000004519 manufacturing process Methods 0.000 claims 5
- 241000590002 Helicobacter pylori Species 0.000 abstract description 15
- 230000000844 anti-bacterial effect Effects 0.000 abstract description 9
- 229960002626 clarithromycin Drugs 0.000 abstract description 5
- AGOYDEPGAOXOCK-KCBOHYOISA-N clarithromycin Chemical compound O([C@@H]1[C@@H](C)C(=O)O[C@@H]([C@@]([C@H](O)[C@@H](C)C(=O)[C@H](C)C[C@](C)([C@H](O[C@H]2[C@@H]([C@H](C[C@@H](C)O2)N(C)C)O)[C@H]1C)OC)(C)O)CC)[C@H]1C[C@@](C)(OC)[C@@H](O)[C@H](C)O1 AGOYDEPGAOXOCK-KCBOHYOISA-N 0.000 abstract description 5
- 238000000605 extraction Methods 0.000 abstract description 5
- GSDSWSVVBLHKDQ-JTQLQIEISA-N Levofloxacin Chemical compound C([C@@H](N1C2=C(C(C(C(O)=O)=C1)=O)C=C1F)C)OC2=C1N1CCN(C)CC1 GSDSWSVVBLHKDQ-JTQLQIEISA-N 0.000 abstract description 4
- 238000011161 development Methods 0.000 abstract description 4
- 238000000338 in vitro Methods 0.000 abstract description 4
- 229960003376 levofloxacin Drugs 0.000 abstract description 4
- VAOCPAMSLUNLGC-UHFFFAOYSA-N metronidazole Chemical compound CC1=NC=C([N+]([O-])=O)N1CCO VAOCPAMSLUNLGC-UHFFFAOYSA-N 0.000 abstract description 4
- 229960000282 metronidazole Drugs 0.000 abstract description 4
- 238000002474 experimental method Methods 0.000 abstract description 3
- 229940124350 antibacterial drug Drugs 0.000 abstract description 2
- 235000013305 food Nutrition 0.000 abstract description 2
- 238000011031 large-scale manufacturing process Methods 0.000 abstract description 2
- 230000003449 preventive effect Effects 0.000 abstract description 2
- 239000001963 growth medium Substances 0.000 description 18
- 230000001580 bacterial effect Effects 0.000 description 15
- 150000001875 compounds Chemical class 0.000 description 15
- 108020004414 DNA Proteins 0.000 description 12
- 238000001514 detection method Methods 0.000 description 10
- 238000004128 high performance liquid chromatography Methods 0.000 description 9
- 239000007788 liquid Substances 0.000 description 9
- 241000228245 Aspergillus niger Species 0.000 description 8
- 230000000694 effects Effects 0.000 description 8
- 235000019439 ethyl acetate Nutrition 0.000 description 7
- 239000000243 solution Substances 0.000 description 7
- 241000894006 Bacteria Species 0.000 description 6
- 238000004440 column chromatography Methods 0.000 description 6
- 239000002609 medium Substances 0.000 description 6
- 239000007787 solid Substances 0.000 description 6
- 238000012258 culturing Methods 0.000 description 5
- 230000002401 inhibitory effect Effects 0.000 description 5
- GBMDVOWEEQVZKZ-UHFFFAOYSA-N methanol;hydrate Chemical compound O.OC GBMDVOWEEQVZKZ-UHFFFAOYSA-N 0.000 description 5
- 239000000203 mixture Substances 0.000 description 5
- 239000003208 petroleum Substances 0.000 description 5
- 239000006187 pill Substances 0.000 description 5
- 238000002953 preparative HPLC Methods 0.000 description 5
- IAZDPXIOMUYVGZ-WFGJKAKNSA-N Dimethyl sulfoxide Chemical compound [2H]C([2H])([2H])S(=O)C([2H])([2H])[2H] IAZDPXIOMUYVGZ-WFGJKAKNSA-N 0.000 description 4
- VYPSYNLAJGMNEJ-UHFFFAOYSA-N Silicium dioxide Chemical compound O=[Si]=O VYPSYNLAJGMNEJ-UHFFFAOYSA-N 0.000 description 4
- HEDRZPFGACZZDS-MICDWDOJSA-N Trichloro(2H)methane Chemical compound [2H]C(Cl)(Cl)Cl HEDRZPFGACZZDS-MICDWDOJSA-N 0.000 description 4
- 239000003242 anti bacterial agent Substances 0.000 description 4
- 238000002347 injection Methods 0.000 description 4
- 239000007924 injection Substances 0.000 description 4
- 239000000463 material Substances 0.000 description 4
- 108090000623 proteins and genes Proteins 0.000 description 4
- 238000010898 silica gel chromatography Methods 0.000 description 4
- 239000000126 substance Substances 0.000 description 4
- 238000012360 testing method Methods 0.000 description 4
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 4
- 241000196324 Embryophyta Species 0.000 description 3
- 230000003321 amplification Effects 0.000 description 3
- 244000052616 bacterial pathogen Species 0.000 description 3
- 230000003115 biocidal effect Effects 0.000 description 3
- 239000003153 chemical reaction reagent Substances 0.000 description 3
- 238000010828 elution Methods 0.000 description 3
- 239000008187 granular material Substances 0.000 description 3
- 230000036541 health Effects 0.000 description 3
- 150000002500 ions Chemical class 0.000 description 3
- 238000002955 isolation Methods 0.000 description 3
- 238000002156 mixing Methods 0.000 description 3
- 238000003199 nucleic acid amplification method Methods 0.000 description 3
- 238000000746 purification Methods 0.000 description 3
- 150000003839 salts Chemical class 0.000 description 3
- 238000000926 separation method Methods 0.000 description 3
- 239000000741 silica gel Substances 0.000 description 3
- 229910002027 silica gel Inorganic materials 0.000 description 3
- 239000002689 soil Substances 0.000 description 3
- 239000000725 suspension Substances 0.000 description 3
- HIXDQWDOVZUNNA-UHFFFAOYSA-N 2-(3,4-dimethoxyphenyl)-5-hydroxy-7-methoxychromen-4-one Chemical compound C=1C(OC)=CC(O)=C(C(C=2)=O)C=1OC=2C1=CC=C(OC)C(OC)=C1 HIXDQWDOVZUNNA-UHFFFAOYSA-N 0.000 description 2
- 229920001817 Agar Polymers 0.000 description 2
- 241000222122 Candida albicans Species 0.000 description 2
- HEDRZPFGACZZDS-UHFFFAOYSA-N Chloroform Chemical compound ClC(Cl)Cl HEDRZPFGACZZDS-UHFFFAOYSA-N 0.000 description 2
- 102000004190 Enzymes Human genes 0.000 description 2
- 108090000790 Enzymes Proteins 0.000 description 2
- 206010019375 Helicobacter infections Diseases 0.000 description 2
- 108091023242 Internal transcribed spacer Proteins 0.000 description 2
- ISWSIDIOOBJBQZ-UHFFFAOYSA-N Phenol Chemical compound OC1=CC=CC=C1 ISWSIDIOOBJBQZ-UHFFFAOYSA-N 0.000 description 2
- 241000191967 Staphylococcus aureus Species 0.000 description 2
- 208000005718 Stomach Neoplasms Diseases 0.000 description 2
- 238000010521 absorption reaction Methods 0.000 description 2
- PBCJIPOGFJYBJE-UHFFFAOYSA-N acetonitrile;hydrate Chemical compound O.CC#N PBCJIPOGFJYBJE-UHFFFAOYSA-N 0.000 description 2
- 230000004913 activation Effects 0.000 description 2
- 239000004480 active ingredient Substances 0.000 description 2
- 239000008272 agar Substances 0.000 description 2
- 238000004458 analytical method Methods 0.000 description 2
- 229940095731 candida albicans Drugs 0.000 description 2
- 239000002775 capsule Substances 0.000 description 2
- 238000007621 cluster analysis Methods 0.000 description 2
- 239000000287 crude extract Substances 0.000 description 2
- 238000010790 dilution Methods 0.000 description 2
- 239000012895 dilution Substances 0.000 description 2
- 239000012153 distilled water Substances 0.000 description 2
- 235000019441 ethanol Nutrition 0.000 description 2
- RTZKZFJDLAIYFH-UHFFFAOYSA-N ether Substances CCOCC RTZKZFJDLAIYFH-UHFFFAOYSA-N 0.000 description 2
- ARXPDHLVDOYIPX-UHFFFAOYSA-N flavasperone Chemical compound O1C(C)=CC(=O)C2=C1C1=C(OC)C=C(OC)C=C1C=C2O ARXPDHLVDOYIPX-UHFFFAOYSA-N 0.000 description 2
- 230000002538 fungal effect Effects 0.000 description 2
- 206010017758 gastric cancer Diseases 0.000 description 2
- 238000004895 liquid chromatography mass spectrometry Methods 0.000 description 2
- 238000004949 mass spectrometry Methods 0.000 description 2
- 238000001819 mass spectrum Methods 0.000 description 2
- 238000012986 modification Methods 0.000 description 2
- 230000004048 modification Effects 0.000 description 2
- VLKZOEOYAKHREP-UHFFFAOYSA-N n-Hexane Chemical compound CCCCCC VLKZOEOYAKHREP-UHFFFAOYSA-N 0.000 description 2
- 239000013642 negative control Substances 0.000 description 2
- 239000008188 pellet Substances 0.000 description 2
- 230000002829 reductive effect Effects 0.000 description 2
- 238000011160 research Methods 0.000 description 2
- 230000000717 retained effect Effects 0.000 description 2
- 230000002441 reversible effect Effects 0.000 description 2
- HFPQKJMLIONCGP-UHFFFAOYSA-N rubrofusarin B Chemical compound O1C(C)=CC(=O)C=2C1=CC1=CC(OC)=CC(OC)=C1C=2O HFPQKJMLIONCGP-UHFFFAOYSA-N 0.000 description 2
- 238000002791 soaking Methods 0.000 description 2
- 239000011734 sodium Substances 0.000 description 2
- 201000011549 stomach cancer Diseases 0.000 description 2
- 238000006467 substitution reaction Methods 0.000 description 2
- 238000005406 washing Methods 0.000 description 2
- XMAYWYJOQHXEEK-OZXSUGGESA-N (2R,4S)-ketoconazole Chemical compound C1CN(C(=O)C)CCN1C(C=C1)=CC=C1OC[C@@H]1O[C@@](CN2C=NC=C2)(C=2C(=CC(Cl)=CC=2)Cl)OC1 XMAYWYJOQHXEEK-OZXSUGGESA-N 0.000 description 1
- 238000001644 13C nuclear magnetic resonance spectroscopy Methods 0.000 description 1
- 108020004463 18S ribosomal RNA Proteins 0.000 description 1
- 238000005160 1H NMR spectroscopy Methods 0.000 description 1
- 238000005084 2D-nuclear magnetic resonance Methods 0.000 description 1
- KKYMYHPILGUDLX-UHFFFAOYSA-N 5H-pyrano[3,2-b]pyrrol-2-one Chemical class N=1C(C=C2C=1C=CCO2)=O KKYMYHPILGUDLX-UHFFFAOYSA-N 0.000 description 1
- TWCMVXMQHSVIOJ-UHFFFAOYSA-N Aglycone of yadanzioside D Natural products COC(=O)C12OCC34C(CC5C(=CC(O)C(O)C5(C)C3C(O)C1O)C)OC(=O)C(OC(=O)C)C24 TWCMVXMQHSVIOJ-UHFFFAOYSA-N 0.000 description 1
- 241000205585 Aquilegia canadensis Species 0.000 description 1
- XOJVHLIYNSOZOO-SWOBOCGESA-N Arctiin Chemical compound C1=C(OC)C(OC)=CC=C1C[C@@H]1[C@@H](CC=2C=C(OC)C(O[C@H]3[C@@H]([C@@H](O)[C@H](O)[C@@H](CO)O3)O)=CC=2)C(=O)OC1 XOJVHLIYNSOZOO-SWOBOCGESA-N 0.000 description 1
- 240000006439 Aspergillus oryzae Species 0.000 description 1
- 235000002247 Aspergillus oryzae Nutrition 0.000 description 1
- PLMKQQMDOMTZGG-UHFFFAOYSA-N Astrantiagenin E-methylester Natural products CC12CCC(O)C(C)(CO)C1CCC1(C)C2CC=C2C3CC(C)(C)CCC3(C(=O)OC)CCC21C PLMKQQMDOMTZGG-UHFFFAOYSA-N 0.000 description 1
- 244000277360 Bruguiera gymnorhiza Species 0.000 description 1
- 238000007400 DNA extraction Methods 0.000 description 1
- 241001360526 Escherichia coli ATCC 25922 Species 0.000 description 1
- 208000007882 Gastritis Diseases 0.000 description 1
- 241000853480 Helicobacter pylori G27 Species 0.000 description 1
- 238000005481 NMR spectroscopy Methods 0.000 description 1
- 241000729876 Niveus Species 0.000 description 1
- 208000008469 Peptic Ulcer Diseases 0.000 description 1
- 239000004698 Polyethylene Substances 0.000 description 1
- 229920002584 Polyethylene Glycol 6000 Polymers 0.000 description 1
- 108010093965 Polymyxin B Proteins 0.000 description 1
- 241000219000 Populus Species 0.000 description 1
- 239000006004 Quartz sand Substances 0.000 description 1
- 240000002044 Rhizophora apiculata Species 0.000 description 1
- 238000012300 Sequence Analysis Methods 0.000 description 1
- GAMYVSCDDLXAQW-AOIWZFSPSA-N Thermopsosid Natural products O(C)c1c(O)ccc(C=2Oc3c(c(O)cc(O[C@H]4[C@H](O)[C@@H](O)[C@H](O)[C@H](CO)O4)c3)C(=O)C=2)c1 GAMYVSCDDLXAQW-AOIWZFSPSA-N 0.000 description 1
- 241000223229 Trichophyton rubrum Species 0.000 description 1
- 108010059993 Vancomycin Proteins 0.000 description 1
- 108010093894 Xanthine oxidase Proteins 0.000 description 1
- 102100033220 Xanthine oxidase Human genes 0.000 description 1
- HCHKCACWOHOZIP-UHFFFAOYSA-N Zinc Chemical compound [Zn] HCHKCACWOHOZIP-UHFFFAOYSA-N 0.000 description 1
- 239000013543 active substance Substances 0.000 description 1
- 238000000246 agarose gel electrophoresis Methods 0.000 description 1
- 238000007605 air drying Methods 0.000 description 1
- 229960003022 amoxicillin Drugs 0.000 description 1
- LSQZJLSUYDQPKJ-NJBDSQKTSA-N amoxicillin Chemical compound C1([C@@H](N)C(=O)N[C@H]2[C@H]3SC([C@@H](N3C2=O)C(O)=O)(C)C)=CC=C(O)C=C1 LSQZJLSUYDQPKJ-NJBDSQKTSA-N 0.000 description 1
- 229940045436 amoxicillin and clarithromycin omeprazole Drugs 0.000 description 1
- 238000000137 annealing Methods 0.000 description 1
- 229940088710 antibiotic agent Drugs 0.000 description 1
- BPYGTFFYYOWDBC-LOVSFRALSA-N arctiin Natural products COc1ccc(C[C@H]2COC(=O)[C@@H]2Cc3ccc(O[C@@H]4O[C@H](C)[C@@H](O)[C@H](O)[C@H]4O)c(OC)c3)cc1OC BPYGTFFYYOWDBC-LOVSFRALSA-N 0.000 description 1
- 125000003118 aryl group Chemical group 0.000 description 1
- 210000004556 brain Anatomy 0.000 description 1
- 239000007853 buffer solution Substances 0.000 description 1
- 238000006555 catalytic reaction Methods 0.000 description 1
- 238000005119 centrifugation Methods 0.000 description 1
- 238000012512 characterization method Methods 0.000 description 1
- 239000003795 chemical substances by application Substances 0.000 description 1
- 208000023652 chronic gastritis Diseases 0.000 description 1
- 238000002983 circular dichroism Methods 0.000 description 1
- 238000001142 circular dichroism spectrum Methods 0.000 description 1
- 239000011248 coating agent Substances 0.000 description 1
- 238000000576 coating method Methods 0.000 description 1
- 229940125904 compound 1 Drugs 0.000 description 1
- 229940125782 compound 2 Drugs 0.000 description 1
- 229940126214 compound 3 Drugs 0.000 description 1
- 229940125898 compound 5 Drugs 0.000 description 1
- 238000005520 cutting process Methods 0.000 description 1
- 230000001472 cytotoxic effect Effects 0.000 description 1
- 238000004925 denaturation Methods 0.000 description 1
- 230000036425 denaturation Effects 0.000 description 1
- 230000001419 dependent effect Effects 0.000 description 1
- 238000010586 diagram Methods 0.000 description 1
- 208000010643 digestive system disease Diseases 0.000 description 1
- 238000007865 diluting Methods 0.000 description 1
- 238000003113 dilution method Methods 0.000 description 1
- 239000007884 disintegrant Substances 0.000 description 1
- 239000012154 double-distilled water Substances 0.000 description 1
- 239000008298 dragée Substances 0.000 description 1
- 239000003480 eluent Substances 0.000 description 1
- 239000012149 elution buffer Substances 0.000 description 1
- 239000002662 enteric coated tablet Substances 0.000 description 1
- 230000008029 eradication Effects 0.000 description 1
- SRCZQMGIVIYBBJ-UHFFFAOYSA-N ethoxyethane;ethyl acetate Chemical compound CCOCC.CCOC(C)=O SRCZQMGIVIYBBJ-UHFFFAOYSA-N 0.000 description 1
- 238000001704 evaporation Methods 0.000 description 1
- 210000000416 exudates and transudate Anatomy 0.000 description 1
- 239000007941 film coated tablet Substances 0.000 description 1
- 229930003944 flavone Natural products 0.000 description 1
- 150000002212 flavone derivatives Chemical class 0.000 description 1
- 235000011949 flavones Nutrition 0.000 description 1
- 230000007760 free radical scavenging Effects 0.000 description 1
- 238000004108 freeze drying Methods 0.000 description 1
- 239000000499 gel Substances 0.000 description 1
- 239000003292 glue Substances 0.000 description 1
- 235000013402 health food Nutrition 0.000 description 1
- 238000010438 heat treatment Methods 0.000 description 1
- PFOARMALXZGCHY-UHFFFAOYSA-N homoegonol Natural products C1=C(OC)C(OC)=CC=C1C1=CC2=CC(CCCO)=CC(OC)=C2O1 PFOARMALXZGCHY-UHFFFAOYSA-N 0.000 description 1
- 238000001802 infusion Methods 0.000 description 1
- 239000003978 infusion fluid Substances 0.000 description 1
- 230000005764 inhibitory process Effects 0.000 description 1
- 230000002262 irrigation Effects 0.000 description 1
- 238000003973 irrigation Methods 0.000 description 1
- MWDZOUNAPSSOEL-UHFFFAOYSA-N kaempferol Natural products OC1=C(C(=O)c2cc(O)cc(O)c2O1)c3ccc(O)cc3 MWDZOUNAPSSOEL-UHFFFAOYSA-N 0.000 description 1
- 229960004125 ketoconazole Drugs 0.000 description 1
- DTOSIQBPPRVQHS-PDBXOOCHSA-M linolenate Chemical compound CC\C=C/C\C=C/C\C=C/CCCCCCCC([O-])=O DTOSIQBPPRVQHS-PDBXOOCHSA-M 0.000 description 1
- 229940040452 linolenate Drugs 0.000 description 1
- 238000004811 liquid chromatography Methods 0.000 description 1
- 229940057995 liquid paraffin Drugs 0.000 description 1
- LRDGATPGVJTWLJ-UHFFFAOYSA-N luteolin Natural products OC1=CC(O)=CC(C=2OC3=CC(O)=CC(O)=C3C(=O)C=2)=C1 LRDGATPGVJTWLJ-UHFFFAOYSA-N 0.000 description 1
- 235000009498 luteolin Nutrition 0.000 description 1
- IQPNAANSBPBGFQ-UHFFFAOYSA-N luteolin Chemical compound C=1C(O)=CC(O)=C(C(C=2)=O)C=1OC=2C1=CC=C(O)C(O)=C1 IQPNAANSBPBGFQ-UHFFFAOYSA-N 0.000 description 1
- 239000008176 lyophilized powder Substances 0.000 description 1
- 230000004060 metabolic process Effects 0.000 description 1
- 239000002207 metabolite Substances 0.000 description 1
- YMWUJEATGCHHMB-UHFFFAOYSA-N methylene chloride Substances ClCCl YMWUJEATGCHHMB-UHFFFAOYSA-N 0.000 description 1
- 230000002906 microbiologic effect Effects 0.000 description 1
- 244000005700 microbiome Species 0.000 description 1
- 210000004400 mucous membrane Anatomy 0.000 description 1
- 229930014626 natural product Natural products 0.000 description 1
- 239000006199 nebulizer Substances 0.000 description 1
- 238000000655 nuclear magnetic resonance spectrum Methods 0.000 description 1
- 108020004707 nucleic acids Proteins 0.000 description 1
- 150000007523 nucleic acids Chemical class 0.000 description 1
- 102000039446 nucleic acids Human genes 0.000 description 1
- 235000015097 nutrients Nutrition 0.000 description 1
- 239000002674 ointment Substances 0.000 description 1
- SBQLYHNEIUGQKH-UHFFFAOYSA-N omeprazole Chemical compound N1=C2[CH]C(OC)=CC=C2N=C1S(=O)CC1=NC=C(C)C(OC)=C1C SBQLYHNEIUGQKH-UHFFFAOYSA-N 0.000 description 1
- 229960000381 omeprazole Drugs 0.000 description 1
- 239000006191 orally-disintegrating tablet Substances 0.000 description 1
- LSQZJLSUYDQPKJ-UHFFFAOYSA-N p-Hydroxyampicillin Natural products O=C1N2C(C(O)=O)C(C)(C)SC2C1NC(=O)C(N)C1=CC=C(O)C=C1 LSQZJLSUYDQPKJ-UHFFFAOYSA-N 0.000 description 1
- 150000007965 phenolic acids Chemical class 0.000 description 1
- 229920000573 polyethylene Polymers 0.000 description 1
- 229940093429 polyethylene glycol 6000 Drugs 0.000 description 1
- 229920001470 polyketone Polymers 0.000 description 1
- 229920000024 polymyxin B Polymers 0.000 description 1
- 229960005266 polymyxin b Drugs 0.000 description 1
- 238000012257 pre-denaturation Methods 0.000 description 1
- 229940126409 proton pump inhibitor Drugs 0.000 description 1
- 239000000612 proton pump inhibitor Substances 0.000 description 1
- 238000011084 recovery Methods 0.000 description 1
- KLKFLNXANXGSIT-UHFFFAOYSA-N rubrofusarin Natural products O1C(C)=CC(=O)C2=C1C=C1C=C(O)C=C(OC)C1=C2O KLKFLNXANXGSIT-UHFFFAOYSA-N 0.000 description 1
- 229930000044 secondary metabolite Natural products 0.000 description 1
- 238000012163 sequencing technique Methods 0.000 description 1
- 210000002966 serum Anatomy 0.000 description 1
- 238000012807 shake-flask culturing Methods 0.000 description 1
- AKHNMLFCWUSKQB-UHFFFAOYSA-L sodium thiosulfate Chemical compound [Na+].[Na+].[O-]S([O-])(=O)=S AKHNMLFCWUSKQB-UHFFFAOYSA-L 0.000 description 1
- 235000019345 sodium thiosulphate Nutrition 0.000 description 1
- 238000001179 sorption measurement Methods 0.000 description 1
- 238000001228 spectrum Methods 0.000 description 1
- 239000007921 spray Substances 0.000 description 1
- 210000002784 stomach Anatomy 0.000 description 1
- 239000007940 sugar coated tablet Substances 0.000 description 1
- 239000003826 tablet Substances 0.000 description 1
- 238000010998 test method Methods 0.000 description 1
- 238000002560 therapeutic procedure Methods 0.000 description 1
- 238000012546 transfer Methods 0.000 description 1
- 230000009466 transformation Effects 0.000 description 1
- 229960001082 trimethoprim Drugs 0.000 description 1
- IEDVJHCEMCRBQM-UHFFFAOYSA-N trimethoprim Chemical compound COC1=C(OC)C(OC)=CC(CC=2C(=NC(N)=NC=2)N)=C1 IEDVJHCEMCRBQM-UHFFFAOYSA-N 0.000 description 1
- 238000002137 ultrasound extraction Methods 0.000 description 1
- 229960003165 vancomycin Drugs 0.000 description 1
- MYPYJXKWCTUITO-UHFFFAOYSA-N vancomycin Natural products O1C(C(=C2)Cl)=CC=C2C(O)C(C(NC(C2=CC(O)=CC(O)=C2C=2C(O)=CC=C3C=2)C(O)=O)=O)NC(=O)C3NC(=O)C2NC(=O)C(CC(N)=O)NC(=O)C(NC(=O)C(CC(C)C)NC)C(O)C(C=C3Cl)=CC=C3OC3=CC2=CC1=C3OC1OC(CO)C(O)C(O)C1OC1CC(C)(N)C(O)C(C)O1 MYPYJXKWCTUITO-UHFFFAOYSA-N 0.000 description 1
- MYPYJXKWCTUITO-LYRMYLQWSA-O vancomycin(1+) Chemical compound O([C@@H]1[C@@H](O)[C@H](O)[C@@H](CO)O[C@H]1OC1=C2C=C3C=C1OC1=CC=C(C=C1Cl)[C@@H](O)[C@H](C(N[C@@H](CC(N)=O)C(=O)N[C@H]3C(=O)N[C@H]1C(=O)N[C@H](C(N[C@@H](C3=CC(O)=CC(O)=C3C=3C(O)=CC=C1C=3)C([O-])=O)=O)[C@H](O)C1=CC=C(C(=C1)Cl)O2)=O)NC(=O)[C@@H](CC(C)C)[NH2+]C)[C@H]1C[C@](C)([NH3+])[C@H](O)[C@H](C)O1 MYPYJXKWCTUITO-LYRMYLQWSA-O 0.000 description 1
- VHBFFQKBGNRLFZ-UHFFFAOYSA-N vitamin p Natural products O1C2=CC=CC=C2C(=O)C=C1C1=CC=CC=C1 VHBFFQKBGNRLFZ-UHFFFAOYSA-N 0.000 description 1
- 239000002699 waste material Substances 0.000 description 1
- 239000003643 water by type Substances 0.000 description 1
- 239000011701 zinc Substances 0.000 description 1
- 229910052725 zinc Inorganic materials 0.000 description 1
Images
Classifications
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K31/00—Medicinal preparations containing organic active ingredients
- A61K31/33—Heterocyclic compounds
- A61K31/335—Heterocyclic compounds having oxygen as the only ring hetero atom, e.g. fungichromin
- A61K31/35—Heterocyclic compounds having oxygen as the only ring hetero atom, e.g. fungichromin having six-membered rings with one oxygen as the only ring hetero atom
- A61K31/352—Heterocyclic compounds having oxygen as the only ring hetero atom, e.g. fungichromin having six-membered rings with one oxygen as the only ring hetero atom condensed with carbocyclic rings, e.g. methantheline
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
- A23L31/00—Edible extracts or preparations of fungi; Preparation or treatment thereof
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
- A23L33/00—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof
- A23L33/10—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof using additives
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P31/00—Antiinfectives, i.e. antibiotics, antiseptics, chemotherapeutics
- A61P31/04—Antibacterial agents
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
- C12N1/14—Fungi; Culture media therefor
-
- 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
- C12P17/16—Preparation of heterocyclic carbon compounds with only O, N, S, Se or Te as ring hetero atoms containing two or more hetero rings
- C12P17/162—Heterorings having oxygen atoms as the only ring heteroatoms, e.g. Lasalocid
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
- A23V2002/00—Food compositions, function of food ingredients or processes for food or foodstuffs
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12R—INDEXING SCHEME ASSOCIATED WITH SUBCLASSES C12C - C12Q, RELATING TO MICROORGANISMS
- C12R2001/00—Microorganisms ; Processes using microorganisms
- C12R2001/645—Fungi ; Processes using fungi
- C12R2001/66—Aspergillus
-
- 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
Landscapes
- Life Sciences & Earth Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Health & Medical Sciences (AREA)
- Engineering & Computer Science (AREA)
- Organic Chemistry (AREA)
- Zoology (AREA)
- Wood Science & Technology (AREA)
- General Health & Medical Sciences (AREA)
- Genetics & Genomics (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Microbiology (AREA)
- Biotechnology (AREA)
- General Engineering & Computer Science (AREA)
- Medicinal Chemistry (AREA)
- Chemical Kinetics & Catalysis (AREA)
- General Chemical & Material Sciences (AREA)
- Mycology (AREA)
- Biochemistry (AREA)
- Polymers & Plastics (AREA)
- Nutrition Science (AREA)
- Veterinary Medicine (AREA)
- Public Health (AREA)
- Animal Behavior & Ethology (AREA)
- Food Science & Technology (AREA)
- Pharmacology & Pharmacy (AREA)
- Epidemiology (AREA)
- Communicable Diseases (AREA)
- Oncology (AREA)
- Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
- Botany (AREA)
- Tropical Medicine & Parasitology (AREA)
- Virology (AREA)
- Biomedical Technology (AREA)
- Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
- Preparation Of Compounds By Using Micro-Organisms (AREA)
Abstract
The invention discloses application of naphtho gamma-pyrone compounds in preparing anti-helicobacter pylori medicaments or prevention health-care products. The invention provides a marine fungus, which is named as Aspergillus sp.DM94, and has a preservation number: CCTCC NO: m20191003. The method extracts and separates the naphtho-gamma-pyrone compounds from the fermentation culture of the marine fungi by utilizing the polarity difference of the naphtho-gamma-pyrone, has simple and convenient operation, high extraction yield and high product purity, and is suitable for large-scale production. In-vitro antibacterial experiments show that the naphtho-gamma-pyrone compound has better in-vitro antibacterial activity, and the MIC value of the naphtho-gamma-pyrone compound is 4-64 mu G/mL aiming at a helicobacter pylori standard strain G27 and a multi-drug-resistant strain HP159 of levofloxacin, clarithromycin and metronidazole, so that the naphtho-gamma-pyrone compound can be used for preparing antibacterial drugs and preventive health-care foods and has good development prospects.
Description
Technical Field
The invention relates to the field of marine fungus metabolites as new antibacterial agents, and in particular relates to application of naphtho gamma-pyrone compounds in preparation of anti-helicobacter pylori medicines or prevention health-care products.
Background
Helicobacter pylori (Helicobacter pylori) is a gram-negative bacterium that is microaerophilic, often located in the mucous membrane of the human stomach fundus, and infects about half of the population worldwide. Helicobacter pylori infection often causes digestive system diseases such as chronic gastritis, peptic ulcer, and the like. Research shows that helicobacter pylori infection is closely related to the occurrence and development of gastric cancer, and the eradication of helicobacter pylori can reduce the risk of gastric cancer. For the clinical treatment of helicobacter pylori, triple or quadruple therapy recommended by relevant guidelines, including omeprazole, a proton pump inhibitor, amoxicillin and clarithromycin, and the like, has good effects. However, the recent clinical emergence of multidrug resistant strains has greatly reduced antibiotic efficacy, and therefore the development of new anti-helicobacter pylori drugs, especially drugs effective against multidrug resistant strains therein, is of great significance.
Aspergillus niger is often capable of metabolizing structurally diverse secondary metabolites as well as abundant enzymes, and is therefore frequently used for bioconversion/catalysis studies of active natural products. Reports on the use of Aspergillus niger have mainly focused on the transformation studies of medicinal plant components using the enzymes produced by Aspergillus niger. For example, in Hwang et al, Aspergillus niger A-T1 is used to convert flavone in plants into phenolic acid components with antibacterial activity [ patent application, 2019, CN 110093279A 20190806], Zhang et al, Aspergillus niger fermentation is used to increase the yield of luteolin in honeysuckle flower [ patent application, 2017, CN 107022590A 20170808], Ou et al, Aspergillus niger ZJUT712 is used to convert arctiin into aglycone [ patent application, 2009, CN 101358173A 20090204 ].
Naphtho-gamma-pyrones are an important class of aromatic polyketones produced by aspergillus fungi, especially aspergillus niger, including naphtho-gamma-pyrones, bis-naphtho-gamma-pyrones, alpha-pyrones, pyrrolopyrones, and the like. The literature reports that the substances show good xanthine oxidase inhibitory activity, and part of compounds have good antibacterial activity on common pathogenic bacteria such as candida albicans, trichophyton rubrum and the like, and the activity of the compounds is related to ketoconazole. Meanwhile, the literature also reports that the substances have free radical scavenging activity, cytotoxic activity and the like. However, no report has been found so far which reports that naphtho- γ -pyrones derived from Aspergillus niger have anti-helicobacter pylori activity.
Disclosure of Invention
The invention aims to provide application of naphtho-gamma-pyrone compounds in preparing anti-helicobacter pylori medicaments or prevention health-care products.
The naphtho-gamma-pyrones compound is a natural active substance with medicinal value extracted from Aspergillus sp.DM94 (DM 94).
The purpose of the invention is realized by the following technical scheme:
the invention provides an application of a naphtho gamma-pyrone compound or pharmaceutically acceptable salt thereof in preparing anti-helicobacter pylori medicaments or prevention health care products, wherein the structural formula of the naphtho gamma-pyrone compound is shown as formulas (1) to (8):
the invention provides a marine fungus, named Aspergillus fungus DM94(Aspergillus sp.DM94), which is an epiphytic fungus of marine plants, and the preservation information is as follows: the preservation unit is as follows: china Center for Type Culture Collection (CCTCC) with a preservation time of 2019, 12 and 3 months and a preservation address: wuhan university, the preservation number: CCTCC NO: m20191003.
The strain of Aspergillus fungus DM94(Aspergillus sp.dm94) has the following microbiological characteristics:
culturing the bacterial colony in a PDA culture medium at 28 ℃ for 5 days, wherein the diameter of the bacterial colony is 35-40 mm, and culturing the bacterial colony for 10 days is 65-70 mm; the colony is initially white, hypha in the center of the colony is turned into black when the colony is cultured for two to three days, and the whole colony is turned into black when the colony is cultured for six days, has white edges and is cotton-shaped in texture and has no exudate; the reverse side of the colony is light yellow. Conidiophores are generated on aerial hyphae, and conidiophores are spherical. The diameter of the spore stems is 10-12.5 mu m, and the wall thickness is smooth; a top capsule is spherical, the diameter is 30-40 mu m, and the surface of the top capsule is fertile; the spore-forming structure has double layers, wherein the stem base is 10-12 multiplied by 2-3 mu m; 6-8 multiplied by 2-3 mu m of bottle stalk; conidiophore spherical, diameter of 3-4 μm, and rough wall.
The Aspergillus fungus DM94(Aspergillus sp.dm94) has the following molecular biological characteristics: ITS and 18S sequences are shown in SEQ ID NO.1 and 2.
The application of the marine fungus in preparing the helicobacter pylori resistant preparation.
The invention also provides a preparation method of the naphtho gamma-pyrone compound, which comprises the following steps:
(1) activating the Aspergillus fungus DM94(Aspergillus sp.DM94), inoculating into a fermentation culture medium, and performing fermentation culture;
(2) after fermentation culture is finished, adding an organic solvent for extraction, separating to obtain an extract, concentrating the extract to obtain an extract, and separating and purifying the extract to obtain the naphtho gamma-pyrone compounds;
in order to better simulate the marine environment and provide sufficient nutrients for the growth and metabolism of microorganisms, the formula of the fermentation medium in the step (1) is preferably as follows:
the formula is as follows based on 50g of corn kernels: 50g corn kernels, 0.86g yeast extract powder, 2.37g ammonium tartrate and MgSO40.17g，KH2PO40.25g, sea salt (2%) 0.4g, ddH2O20 mL, and natural PH; the corn grains need to be crushed into particles.
Or, the formula is as follows according to the volume of 1L: 1-5 g of starch, 10-20 g of bran, 3-15 g of yeast extract and KH2PO4 1～8g，MgSO4·7H20.1-0.8 g of O, and the balance being seawater;
or, based on 1L of the volume, the formula is as follows: 200-600 g of potatoes, 2-10 g of peptone, 1-5 g of yeast extract, 5-20 g of glucose and the balance of seawater; the initial pH value is 6.0-7.0;
or, the formula is as follows according to the volume of 1L: 10-40 g of cane sugar, 5-20 g of corn flour and NaNO31-4 g, 1-4 g yeast extract and KH2PO4 0.2～0.8g、MgSO4·7H2O 0.2～1g、KCl 0.2～1g、FeSO40.001-0.005 g, and the balance of artificial seawater.
Preferably, the fermentation culture condition in the step (1) is static culture at 20-30 ℃ for 10-40 days. The static culture mode is not to carry out shake flask culture.
Further, the condition of fermentation culture is static culture at 22-26 ℃ for 10-40 days. More preferably, the naphtho-gamma-pyrone compound is statically cultured at 25 ℃ for 20 days, under which the yield of the naphtho-gamma-pyrone compound is the highest.
Preferably, the organic solvent in step (2) is one or two of methanol, ethanol, ethyl acetate and acetone.
Preferably, in the step (2), the extract obtained by leaching the fermentation product is subjected to normal phase silica gel column chromatography and then to reverse phase silica gel column chromatography/high performance liquid chromatography. The method specifically adopts silica gel open column chromatography, ODS column chromatography, preparative HPLC and other methods for separation and purification.
When normal phase silica gel column chromatography is adopted for separation, the eluent can adopt one or more of petroleum ether, n-hexane, ethyl acetate, alcohol and water for gradient elution of the extract.
Preferably, the separation and purification comprises: separating the extract by normal phase silica gel column chromatography, gradient eluting with petroleum ether/ethyl acetate mixture and ethyl acetate at volume ratio of 100:0, 9:1, 8:2, 6:4, and 0:100, and collecting each eluted fraction.
The research of the invention shows that the naphtho-gamma-pyrone compound separated from the marine fungus fermentation culture by the method has better antibacterial activity, so the invention also provides the application of the naphtho-gamma-pyrone compound in preparing anti-helicobacter pylori medicines or prevention health products.
The drug is prepared by taking the naphtho-gamma-pyrone compound or the pharmaceutically acceptable salt thereof as a main active ingredient and adding pharmaceutically acceptable auxiliary materials, and can be prepared into a preparation according to a preparation method recorded in pharmaceutics. The preparation can be injection, infusion solution, powder for injection, granule, tablet, granule, powder, oral liquid, sugar-coated tablet, film-coated tablet, enteric-coated tablet, buccal agent, granule, pill, unguent, pellet, spray, dripping pill, disintegrant, orally disintegrating tablet, pellet, etc.
The health product is prepared by taking the naphtho-gamma-pyrone compound or the pharmaceutically acceptable salt thereof as a main active ingredient and adding acceptable health food auxiliary materials.
Compared with the prior art, the invention has the following advantages and effects:
(1) the invention extracts and separates the naphtho-gamma-pyrone compounds from the fermentation culture of marine fungi by utilizing the polarity difference of the naphtho-gamma-pyrone, and the method has simple and convenient operation, high extraction yield and high product purity and is suitable for large-scale production.
(2) In-vitro antibacterial experiments show that the naphtho gamma-pyrone compound provided by the invention has better in-vitro antibacterial activity, and the MIC value of the naphtho gamma-pyrone compound is 4-64 mu G/mL aiming at a helicobacter pylori standard strain (G27) and a multi-drug-resistant strain HP159 of levofloxacin, clarithromycin and metronidazole, so that the naphtho gamma-pyrone compound can be used for preparing antibacterial drugs and preventive health-care foods and has good development prospects.
Drawings
FIG. 1 is a structural formula of naphtho gamma-pyrones of the present invention.
FIG. 2 is HPLC detection spectrum of crude fermentation extract of strain DM 94.
FIG. 3 is a graph of ITS gene sequence cluster analysis of strain DM 94.
FIG. 4 is a diagram of 18S rRNA gene sequence cluster analysis of strain DM 94.
Detailed Description
The present invention will be described in further detail with reference to examples and drawings, but the present invention is not limited thereto.
The test methods in the following examples, in which specific experimental conditions are not specified, are generally performed according to conventional experimental conditions or according to the experimental conditions recommended by the manufacturer. The materials, reagents and the like used are, unless otherwise specified, reagents and materials obtained from commercial sources.
Example 1 isolation of fungus DM94
The strain DM94 is separated from root soil of Bruguiera gymnorrhiza of mangrove, the soil sample is put in a sterile culture dish, air-dried in natural environment, 1g of air-dried soil sample is weighed, 10mL of 50% aged seawater is used for dissolving to obtain 10-1Solution, further diluting to 10-2、10-3Obtaining three gradient sample liquids. Coating 100 μ L of the extract on GPY culture medium, Martin culture medium, CA culture medium, and PDA culture mediumAnd 2 isolation culture media with different dilution concentrations are used in parallel on the SDA culture medium and the fungus No. 2 culture medium, and are cultured for 1-8 weeks at 28 ℃. And observing and picking a single colony, and inoculating the single colony on a No. 2 fungus culture medium to obtain a pure culture strain.
Example 2 molecular characterization of fungal DM94
Extraction of genomic DNA: the DM94 genome DNA template is obtained by a nucleic acid rapid extractor, and the operation is as follows: taking a sterile DNA extraction tube filled with quartz sand, adding 500 μ L of TE, picking an appropriate amount of mycelia, placing into the tube, and extracting for 1min with high speed shaking. Then, 500. mu.L of phenol/chloroform solution was added thereto, and the mixture was centrifuged to obtain genomic DNA.
Amplification of the gene sequence: mu.L of each of 2-fold PCR mastermix 12.5. mu.L, primers ITS1(5'-TCC GTA GGT GAA CCT GCG G-3') and ITS4(5'-TCC TCC GCT TAT TGA TAT GC-3'), (5'-AAC CTG GTT GAT CCT GCC AGT-3' for 18SF and 5'-CGA CGG GCG GTG TGT AC-3' for 18 SR), 2. mu.L of template DNA, and 13.5. mu.L of double distilled water were added. The amplification condition is pre-denaturation at 95 ℃ for 3 min; denaturation at 95 deg.C for 30S, annealing at 59 deg.C for 30S, and extension at 72 deg.C for 30S for 32 cycles; after extension at 72 ℃ for 10 min. The PCR product was detected by 2% agarose gel electrophoresis.
Cutting a target band (about 600bp) for recovery and purification, adding a buffer solution to submerge agar, and carrying out water bath at 42 ℃ until the gel is melted; transferring the glue solution into a collecting tube, and centrifuging to remove waste liquid; washing twice with a washing solution; opening the cover and centrifuging for 1min to volatilize alcohol; the adsorption column is replaced into a new centrifuge tube, 40 mu L of elution buffer solution is added for centrifugation to obtain an amplification product DNA, and the DNA is sent to Huada gene sequencing.
And (3) submitting ITS sequences and 18S sequences to GeneBank for comparison, selecting sequences with higher similarity, carrying out sequence analysis by using Bioedit, and then constructing a phylogenetic tree by using MEGA 7.0. The phylogenetic tree was used to infer that the strain belongs to Aspergillus (FIGS. 3 and 4).
The fungus DM94, designated Aspergillus sp 94(Aspergillus sp.dm94), deposited information: the preservation unit: china Center for Type Culture Collection (CCTCC) with a preservation time of 2019, 12 and 3 months and a preservation address: wuhan university, the preservation number: CCTCC NO: m20191003.
EXAMPLE 3 Mini-preparation of crude fermentation extract of fungus DM94
The strain is cultured in corn solid fermentation medium (corn grain 50 g; ammonium tartrate 2.37 g; yeast extract powder 0.86 g; KH)2PO40.25g；MgSO40.17 g; sea salt 0.4 g; 20mL of distilled water, and natural pH. (Hongkudu, Populus niveus, a high-yielding pseudoshell compound strain Aspergillus oryzae TKYX429 and its application, ZL201310186483.4)) was cultured at 28 ℃ for 4 weeks. Soaking the fermentation product in 80% acetone for 12h, performing ultrasonic extraction for 20min with an ultrasonic extractor, extracting with ethyl acetate for 3-5 times, mixing the extractive solutions, evaporating to dryness, and dissolving with chromatographic methanol. The sample is used for antibacterial activity detection and HPLC detection.
EXAMPLE 4 preliminary examination of antibacterial Activity of fermentation crude extract
The 3 pathogenic bacteria Escherichia coli ATCC 25922, Staphylococcus aureus ATCC 5165 and Candida albicans ATCC 10231 were cultured overnight to prepare 1070.1mL of cfu/mL of bacterial suspension is uniformly coated on a culture medium, after air drying, 20 mu L of crude fermentation extract dissolved in methanol is added on a sterile filter paper sheet (diameter is 6mm), and the sterile filter paper sheet is attached to a flat plate, and pure methanol is used as a blank control. Culturing for 12-24 h at a certain temperature, and observing and measuring the inhibition zone. The detection result shows that the crude fermentation extract of the strain DM94 has the effect of inhibiting staphylococcus aureus and has no effect on other two pathogenic bacteria.
The minimum inhibitory concentration (MIC, 100 mu.L system) of the fungus DM94 fermented extract on helicobacter pylori is detected by a microdilution method. The H.pylori standard strain G27 and the levofloxacin and clarithromycin and metronidazole multidrug-resistant strain (HP159) were provided by the subject group of professor Bighouka of Nanjing medical university. Preparation of MIC plate: 173.6 mu L of culture medium is firstly added into the first hole, and then 6.4 mu L of antibacterial agent is added, and the mixture is diluted to the 7 th hole in a multiple ratio; no drug was added to well 8, and 90. mu.L of the medium was retained as a control with and without drug added. Preparing bacterial liquid: taking helicobacter pylori growing in logarithmic phase on a solid plate, preparing into bacterial suspension by using BHI culture medium, adjusting concentration OD600Is 0.3 (1X 10)8CFU/mL), 10-fold dilution at 1X 107CFU/mL, spare. Inoculating a bacterial liquid:adding 10 μ L into 1-8 th well (the concentration of bacteria liquid in each well is about 1.0 × 10)6CFU/mL), and culturing for 72h to judge the result. The drug concentrations in the 1 st to 7 th wells are 128, 64, 32, 16, 8, 4 and 2 mug/mL respectively. And (5) judging a result: the MIC was taken as the lowest drug concentration that completely inhibited bacterial growth in the wells. The test is only meaningful when the bacteria grow significantly in the 8 th well (i.e., no antibiotic) of the negative control well. When a single jump hole occurs in the microdilution method, the highest drug concentration that inhibits bacterial growth should be recorded. If multiple jump holes appear, the result should not be reported, and the test needs to be repeated. Each drug was tested in 3 replicates. The results show that the MIC value of the fungal DM94 fermented extract against the helicobacter pylori standard strain G27 is 128. mu.g/mL.
EXAMPLE 5 HPLC and LC-MS detection of crude fermentation extract
HPLC detection conditions: and (3) chromatographic column: sinochrom ODS-BP, 250X 4.6nm, 5 μm; the mobile phase is mixed with methanol: the water ratio is respectively 10: 90, respectively; 100: 0; 10: and eluting for 90 min to 15min, 15min to 20min and 20min to 25 min. The detection wavelength is 190-400 nm, and the specified wavelength is 254 nm; the amount of the sample was 20. mu.L.
The LC-MS detection conditions are as follows: ThermoFisher column, 150X 2.1mm, 3 μm; the detection wavelength is 254nm, and the sample injection amount is 10 mu L. Mass spectrometry conditions: positive ion mode: ion source ESI, heating temperature 400 ℃, sheath gas flow 30psi, auxiliary gas flow 5psi, nebulizer voltage 4KV, ion transfer tube temperature 325 ℃. Event one: full scanning, wherein the range is 200-2000 m/z; and event two: data Dependent Scan, activation type CID, minimum signal strength 50000, isolation width 2, collision energy 35, charge state 1, activation time 30 ms.
The detection result shows that the DM94 fermented crude extract has a series of absorption (figure 2) in liquid chromatography for 13-18 min, and the maximum absorption peaks are 307nm and 279 nm. Find [ M + H in Mass Spectrometry]+、[M+Na]+、[2M+H]+、[2M+Na]+As a compound, comparison of at least two of the data in the database of Antibase revealed that 28 compounds could be found in the crude fermentation extract, half of which were unknown, indicating new compounds in the product.
EXAMPLE 6 Mass fermentation culture of the fungus DM94
Placing fungus DM94 in GPY solid medium (glucose 20g, peptone 10g, yeast extract 10g, sea salt 15g, agar 20g, ddH)2O1L, pH7.5), in corn solid medium (50g corn kernel, yeast extract powder 0.86g, ammonium tartrate 2.37g, MgSO4 0.17g，KH2PO40.25g, sea salt (2%) 0.4g, ddH2O20 mL, natural pH) was used to ferment a large amount of 1.5 Kg. After 24 days of fermentation at room temperature, the mycelium was soaked overnight with EtOAc for extraction.
EXAMPLE 7 preparation of naphtho-gamma-pyrones
Soaking and extracting a fermentation product of marine fungus DM94 fermented by a corn solid culture medium for 3-5 times by using ethyl acetate (EtOAC). Mixing extractive solutions, and concentrating under reduced pressure to obtain 50g extract. The extract was subjected to silica gel open column chromatography (phi 50X 640mm, 200-300 mesh, 500-700 g), and gradient eluted with petroleum ether-ethyl acetate (100: 0, 98: 2, 95: 5, 9:1, 8:2, 6:4, 0:100) and EtOAC-MeOH (95: 5, 9: 1) to give 11 fractions (Fr.1-Fr.11). Wherein Fr.7[0.2634g, Petroleum/EtOAC (8:2)]By preparative HPLC (YMC-Pack ODS-A, 80% MeOH-H)2O, 3ml/min) to give compound 1(4.0mg) and compound 2(4.8 mg); fr.9[21.8949g, Petroleum/EtOAC (0:100)]Performing ODS open column chromatography (phi 45X 500mm), and gradient eluting with methanol-water (20% -100%) to obtain 21 sub-fractions (Fr.9-1-Fr.9-21). Subfraction Fr.9-11(2.34g, 70% MeOH-H)2O) is subjected to silica gel open column chromatography (phi 25 is multiplied by 130mm, 200 to 300 meshes, 14g) and CH2Cl2Gradient elution with MeOH (100: 0, 100: 1, 100: 2, 100: 4, 0:100) yielded 6 sub-fractions (Fr.9-11-1 to Fr.9-11-6). Fr.9-11-3[ (213.6mg, CH)2Cl2/MeOH(100：1)]By preparative HPLC (YMC-Pack ODS-A, 63% MeOH-H)2O, 3ml/min) to obtain 8 sub-fractions (Fr.9-11-3-1 to Fr.9-11-3-8). Wherein Fr.9-11-3-5(9.6mg, 63% MeOH-H)2O) preparation by two HPLC (YMC-Pack ODS-A, 63% MeOH-H2O, 3ml/min) to obtain compound 6(1.1mg) and Fr.9-11-3-5-1(3.7 mg); fr.9-11-3-5-1(3.7mg) was further prepared by HPLC (YMC-Pack ODS-A, 60% MeOH-H)2O, 3ml/min) to give compound 7(3.2 mg). Fr.9-11-3-7(47.1 mg)，63％MeOH-H2O) preparation by HPLC (YMC-Pack ODS-A, 63% MeOH-H2O, 3ml/min) gave compound 8(10.6 mg). Subfraction Fr.9-12/13(1.18g, 70% MeOH-H)2O) is subjected to ODS medium-low pressure column chromatography (phi 25 is multiplied by 90mm, 12g) and methanol-water (40-100%) gradient elution to obtain 10 sub-fractions (Fr.9-12/13-1-Fr.9-12/13-10), wherein Fr.9-12/13-5(68.2mg, 60% MeOH-H)2O) by preparative HPLC (YMC-Pack ODS-A, 45% ACN-H2O, 3ml/min) to give compound 5(1.5 mg). Sub-fraction Fr.9-14/15(2.51g, 70% MeOH-H)2O) by preparative HPLC (YMC-Pack ODS-A, 50% ACN-H2O, 3ml/min) to give compound 3(12.8mg) and compound 4(14 mg).
Example 8 structural identification of naphtho-gamma-pyrones
Identifying the purity of the prepared compound by TLC and HPLC, identifying the structure of a sample with the purity of more than 98% by mass spectrum, nuclear magnetic resonance and CD spectrum, measuring the nuclear magnetic resonance by a Bruker AV-600NMR spectrometer, and using TMS as an internal standard; the high-resolution mass spectrum is measured by a Waters Synapt G2 mass spectrometer; the CD spectrometer was a JASCO J-810 circular dichroism spectrometer.
From the results of one-dimensional and two-dimensional NMR analyses of the compounds (see tables 1 and 2), it can be seen that the structures of the compounds were determined as shown in fig. 1.
TABLE 1 preparation of naphtho gamma-pyrones 1 to 813C NMR data
Wherein the content of the first and second substances,ain DMSO-d6 bin CDCl3。
TABLE 2 preparation of naphtho-gamma-pyrones 1-81H NMR data
Wherein the content of the first and second substances,ain DMSO-d6 bin CDCl3。
example 9 analysis of anti-helicobacter pylori Activity of naphtho-gamma-pyrones
9.1 helicobacter pylori strain information: standard strain G27; levofloxacin and clarithromycin and metronidazole multiple drug resistant strain (HP159) (provided by professor topic group of chunkka university of south beijing medical science). The bacterial strains are also disclosed in the patent CN201810462768.9, the preparation of zinc linolenate and the application thereof in preparing the anti-helicobacter pylori medicaments.
9.2 culture medium and main reagents, consumables: columbia culture medium, brain heart infusion culture medium, selective antibiotics (vancomycin, polymyxin B and trimethoprim), serum, omeprazole, amoxicillin and clarithromycin; EP tubes, Tip heads, centrifuge tubes, polyethylene 96-well plates, and the like.
9.3 Experimental methods: the minim dilution method is used for detecting the minimum inhibitory concentration (MIC, 100 mu L system) of the drug to helicobacter pylori, and the MIC plate is prepared: 173.6 mu L of culture medium is firstly added into the first hole, and then 6.4 mu L of antibacterial agent is added, and the mixture is diluted to the 7 th hole by times; no drug was added to well 8, and 90. mu.L of the medium was retained as a control for adding bacteria and no drug.
Preparing bacterial liquid: taking helicobacter pylori growing in logarithmic phase on solid plate, preparing into bacterial suspension with BHI culture medium, adjusting concentration OD600 to 0.3(1 × 10)8CFU/mL), diluted 10-fold to 1X 107CFU/mL, spare.
Inoculating a bacterial liquid: adding 10 μ L into 1-8 th well (the concentration of bacteria liquid in each well is about 1.0 × 10)6CFU/mL). Culturing for 72h to judge the result. The drug concentrations in the 1 st to 7 th wells are 64, 32, 16, 8, 4, 2 and 1. mu.g/mL respectively.
9.4 judging the result: the lowest drug concentration that completely inhibited bacterial growth in the wells was taken as the MIC. The test is only meaningful when the bacteria grow significantly in the 8 th well (i.e., no antibiotic) of the negative control well. When a single jump hole occurs in the microdilution method, the highest concentration of drug that inhibits bacterial growth should be recorded. If a plurality of jump holes appear, the result should not be reported, and the test needs to be repeated. Each drug was tested in 3 replicates.
TABLE 3 MIC (μ g/mL) for naphtho γ -pyrone compounds for inhibiting helicobacter pylori
| Compounds | H.pylori G27 | H.pylori HP159 |
| Flavasperone(1) | ＞64 | ＞64 |
| Rubrofusarin B(2) | ＞64 | ＞64 |
| S-Asperpyrone C(3) | 64 | 64 |
| S-Aurasperone A(4) | 8 | 16 |
| S-Fonsecinone A(5) | ＞64 | ＞64 |
| S-Asperpyrone A(6) | 4 | 4 |
| S-Aurasperone B(7) | 8 | 16 |
| S-Aurasperone F(8) | 4 | 4 |
The results are shown in Table 3, which shows that part of naphtho gamma- pyrone compounds 3, 4 and 6-8 derived from Aspergillus fungus DM94 show good anti-helicobacter pylori activity, and MIC values of the naphtho gamma-pyrone compounds for helicobacter pylori common strain G27 and drug-resistant strain HP159 are both 4-64 mu G/mL.
EXAMPLE 10 preparation of dropping pill preparation containing naphtho-gamma-pyrone compounds
0.5g of naphtho gamma-pyrone compound and 10.5g of polyethylene glycol-6000 are uniformly mixed, heated and melted, the mixture is moved into dripping pills for drip irrigation, the medicine is dripped into liquid paraffin with the temperature of 6-8 ℃, oil is removed, and 300 dripping pills are prepared.
EXAMPLE 11 preparation of lyophilized powder for injection containing naphtho-gamma-pyrone compounds
Taking 0.5g of naphtho gamma-pyrone compounds, 4.5g of glucose, 0.9g of sodium thiosulfate and 1000mL of distilled water, uniformly mixing the components, freeze-drying, and subpackaging 400.
The above embodiments are preferred embodiments of the present invention, but the present invention is not limited to the above embodiments, and any other changes, modifications, substitutions, combinations, and simplifications which do not depart from the spirit and principle of the present invention should be construed as equivalents thereof, and all such changes, modifications, substitutions, combinations, and simplifications are intended to be included in the scope of the present invention.
Sequence listing
<110> river-south university
Application of <120> naphtho gamma-pyrone compounds in preparation of anti-helicobacter pylori medicines or prevention health-care products
<160> 6
<170> SIPOSequenceListing 1.0
<210> 1
<211> 618
<212> DNA
<213> Artificial Sequence (Artificial Sequence)
<220>
<223> ITS sequence of Aspergillus fungus DM94(Aspergillus sp. DM94)
<400> 1
caaaatttcc ttccgctttt tgatatgctt aagttcagcg ggtatcccta cctgatccga 60
ggtcaacctg gaaagaatgg ttggaaaacg tcggcaggcg ccggccaatc ctacagagca 120
tgtgacaaag ccccatacgc tcgaggatcg gacgcggtgc cgccgctgcc tttcgggccc 180
gtccccccgg agagggggac ggcgacccaa cacacaagcc gggcttgagg gcagcaatga 240
cgctcggaca ggcatgcccc ccggaatacc agggggcgca atgtgcgttc aaagactcga 300
tgattcactg aattctgcaa ttcacattag ttatcgcatt tcgctgcgtt cttcatcgat 360
gccggaacca agagatccat tgttgaaagt tttaactgat tgcattcaat caactcagac 420
tgcacgcttt cagacagtgt tcgtgttggg gtctccggcg ggcacgggcc cggggggcag 480
aggcgccccc ccggcggccg acaagcggcg ggcccgccga agcaacaggg tacaatagac 540
acggatggga ggttgggccc aaaggacccg cactcggtaa tgatccttcc gaggcccccc 600
cttacggaag cttacgga 618
<210> 2
<211> 1576
<212> DNA
<213> Artificial Sequence (Artificial Sequence)
<220>
<223> 18S sequence of Aspergillus fungus DM94(Aspergillus sp. DM94)
<400> 2
gactgtctag attagccatg catgtctaag tataagcact ttatactgtg aaactgcgaa 60
tggctcatta aatcagttat cgtttatttg atagtacctt actacatgga tacctgtggt 120
aattctagag ctaatacatg ctgaaaacct cgacttcgga aggggtgtat ttattagata 180
aaaaaccaat gcccttcggg gctccttggt gaatcataat aacttaacga atcgcatggc 240
cttgcgccgg cgatggttca ttcaaatttc tgccctatca actttcgatg gtaggatagt 300
ggcctaccat ggtggcaacg ggtaacgggg aattagggtt cgattccgga gagggagcct 360
gagaaacggc taccacatcc aaggaaggca gcaggcgcgc aaattaccca atcccgacac 420
ggggaggtag tgacaataaa tactgatacg gggctctttt gggtctcgta attggaatga 480
gtacaatcta aatcccttaa cgaggaacaa ttggagggca agtctggtgc cagcagccgc 540
ggtaattcca gctccaatag cgtatattaa agttgttgca gttaaaaagc tcgtagttga 600
accttgggtc tggctggccg gtccgcctca ccgcgagtac tggtccggct ggacctttcc 660
ttctggggaa tctcatggcc ttcactggct gtggggggaa ccaggacttt tactgtgaaa 720
aaattagagt gttcaaagca ggcctttgct cgaatacatt agcatggaat aatagaatag 780
gacgtgcggt tctattttgt tggtttctag gaccgccgta atgattaata gggatagtcg 840
ggggcgtcag tattcagctg tcagaggtga aattcttgga tttgctgaag actaactact 900
gcgaaagcat tcgccaagga tgttttcatt aatcagggaa cgaaagttag gggatcgaag 960
acgatcagat accgtcgtag tcttaaccat aaactatgcc gactagggat cggacggtgt 1020
ttctattatg acccgttcgg caccttacga gaaatcaaag tttttgggtt ctggggggag 1080
tatggtcgca aggctgaaac ttaaagaaat tgacggaagg gcaccaccag gcgtggagcc 1140
tgcggcttaa tttgactcaa cacggggaaa ctcaccaggt ccagacaaaa taaggattga 1200
cagattgaga gctctttctt gatcttttgg atggtggtgc atggccgttc ttagttggtg 1260
gagtgatttg tctgcttaat tgcgataacg aacgagacct cggcccttaa atagcccggt 1320
ccgcatttgc gggccgctgg cttcttaggg ggactatcgg ctcaagccga tggaagtgcg 1380
cggcaataac aggtctgtga tgcccttaga tgttctgggc cgcacgcgcg ctacactgac 1440
agggccagcg agtacatcac cttggccgag aggtctgggt aatcttgtta aaccctgtcg 1500
tgctggggat agagcattgc aattattgct cttcaacgag gaatgcctag taggcacgat 1560
catcagctct ccaatg 1576
<210> 3
<211> 19
<212> DNA
<213> Artificial Sequence (Artificial Sequence)
<220>
<223> ITS1
<400> 3
tccgtaggtg aacctgcgg 19
<210> 4
<211> 20
<212> DNA
<213> Artificial Sequence (Artificial Sequence)
<220>
<223> ITS4
<400> 4
tcctccgctt attgatatgc 20
<210> 5
<211> 21
<212> DNA
<213> Artificial Sequence (Artificial Sequence)
<220>
<223> 18SF
<400> 5
aacctggttg atcctgccag t 21
<210> 6
<211> 17
<212> DNA
<213> Artificial Sequence (Artificial Sequence)
<220>
<223> 18SR
<400> 6
cgacgggcgg tgtgtac 17
Claims (9)
1. A marine fungus characterized by: the Aspergillus sp 94(Aspergillus sp.dm94) is preserved in the China center for type culture collection of Wuhan university in 2019, 12 months and 3 days, with the preservation number: CCTCC NO: m20191003.
2. Use of the marine fungus of claim 1 for the preparation of naphtho γ -pyrones.
3. A preparation method of naphtho gamma-pyrone compounds is characterized by comprising the following steps: the method comprises the following steps:
(1) activating the Aspergillus DM94 of claim 1, inoculating into a fermentation medium, and performing fermentation culture;
(2) after the fermentation culture is finished, adding an organic solvent for leaching, separating to obtain a leaching solution, concentrating the leaching solution to obtain an extract, and separating and purifying the extract to obtain a naphtho gamma-pyrone compound;
the structural formula of the naphtho gamma-pyrone compound is shown in formulas (1) to (8):
4. the production method according to claim 3, characterized in that:
the formula of the fermentation medium in the step (1) is as follows:
the formula is as follows based on 50g of corn kernels: 50g corn kernels, 0.86g yeast extract powder, 2.37g ammonium tartrate and MgSO4 0.17g，KH2PO40.25g, 2% sea salt 0.4g, ddH2O20 mL, pH is natural; the corn grains need to be crushed into particles.
5. The production method according to claim 3, characterized in that:
the formula of the fermentation medium in the step (1) is as follows:
the formula is as follows based on the volume of 1L: 1-5 g of starch, 10-20 g of bran, 3-15 g of yeast extract and KH2PO4 1～8g，MgSO4·7H20.1-0.8 g of O, and the balance being seawater.
6. The production method according to claim 3, characterized in that:
the formula of the fermentation medium in the step (1) is as follows:
the formula is as follows by volume 1L: 200-600 g of potatoes, 2-10 g of peptone, 1-5 g of yeast extract, 5-20 g of glucose and the balance of seawater; the initial pH value is 6.0-7.0.
7. The production method according to claim 3, characterized in that:
the formula of the fermentation medium in the step (1) is as follows:
the formula is as follows by volume 1L: 10-40 g of cane sugar, 5-20 g of corn flour and NaNO31-4 g, 1-4 g yeast extract and KH2PO40.2～0.8g、MgSO4·7H2O 0.2～1g、KCl 0.2～1g、FeSO40.001-0.005 g, and the balance of artificial seawater.
8. The production method according to any one of claims 3 to 7, characterized in that:
the condition of fermentation culture in the step (1) is static culture at 20-30 ℃ for 10-40 days;
the organic solvent in the step (2) is one or two of methanol, ethanol, ethyl acetate and acetone.
9. The method of claim 8, wherein:
the condition of fermentation culture is static culture at 22-26 ℃ for 10-40 days.
Priority Applications (2)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202210605250.2A CN114848628B (en) | 2020-03-26 | 2020-03-26 | Application of naphtho gamma-pyrone compounds in preparation of anti-helicobacter pylori medicines or health care products |
| CN202010223692.1A CN113444644B (en) | 2020-03-26 | 2020-03-26 | Application of naphtho-gamma-pyrone compounds in preparation of anti-helicobacter pylori medicines or prevention health-care products |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202010223692.1A CN113444644B (en) | 2020-03-26 | 2020-03-26 | Application of naphtho-gamma-pyrone compounds in preparation of anti-helicobacter pylori medicines or prevention health-care products |
Related Child Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN202210605250.2A Division CN114848628B (en) | 2020-03-26 | 2020-03-26 | Application of naphtho gamma-pyrone compounds in preparation of anti-helicobacter pylori medicines or health care products |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN113444644A CN113444644A (en) | 2021-09-28 |
| CN113444644B true CN113444644B (en) | 2022-07-19 |
Family
ID=77807618
Family Applications (2)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN202010223692.1A Active CN113444644B (en) | 2020-03-26 | 2020-03-26 | Application of naphtho-gamma-pyrone compounds in preparation of anti-helicobacter pylori medicines or prevention health-care products |
| CN202210605250.2A Active CN114848628B (en) | 2020-03-26 | 2020-03-26 | Application of naphtho gamma-pyrone compounds in preparation of anti-helicobacter pylori medicines or health care products |
Family Applications After (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN202210605250.2A Active CN114848628B (en) | 2020-03-26 | 2020-03-26 | Application of naphtho gamma-pyrone compounds in preparation of anti-helicobacter pylori medicines or health care products |
Country Status (1)
| Country | Link |
|---|---|
| CN (2) | CN113444644B (en) |
Families Citing this family (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN114292254B (en) * | 2021-12-28 | 2023-10-20 | 浙江工业大学 | Tetrahydroxanthone dimer compound and preparation method and application thereof |
Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2009119667A1 (en) * | 2008-03-27 | 2009-10-01 | 国立大学法人広島大学 | Helicobacter pylori GROWTH INHIBITOR AND METHOD FOR PRODUCTION THEREOF |
| CN108383824A (en) * | 2018-04-09 | 2018-08-10 | 陕西科技大学 | A kind of benzopyrone dimer and its extracting method and application |
| WO2020017864A1 (en) * | 2018-07-19 | 2020-01-23 | 한국해양과학기술원 | Protein tyrosine phosphatase 1b inhibitor from antarctic marine-derived aspergillus sp. sf-5929 |
Family Cites Families (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN106434783B (en) * | 2016-04-05 | 2019-08-27 | 广东工业大学 | A kind of chromone compound and preparation method thereof and the application in preparation antibacterials |
| CN108148031B (en) * | 2018-02-06 | 2020-04-28 | 中国海洋大学 | Pyrone compound, preparation method and application |
| CN110172410A (en) * | 2019-05-21 | 2019-08-27 | 扬州大学 | A kind of aphthopyrans ketone compound in marine fungi source, preparation method and applications |
-
2020
- 2020-03-26 CN CN202010223692.1A patent/CN113444644B/en active Active
- 2020-03-26 CN CN202210605250.2A patent/CN114848628B/en active Active
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2009119667A1 (en) * | 2008-03-27 | 2009-10-01 | 国立大学法人広島大学 | Helicobacter pylori GROWTH INHIBITOR AND METHOD FOR PRODUCTION THEREOF |
| CN108383824A (en) * | 2018-04-09 | 2018-08-10 | 陕西科技大学 | A kind of benzopyrone dimer and its extracting method and application |
| WO2020017864A1 (en) * | 2018-07-19 | 2020-01-23 | 한국해양과학기술원 | Protein tyrosine phosphatase 1b inhibitor from antarctic marine-derived aspergillus sp. sf-5929 |
Non-Patent Citations (6)
| Title |
|---|
| 7-Pyrone Compoundswith Selective and Potent Anti-Helicobacter pylori Activity;Masatoshi Taniguchi等;<THE JOURNAL OF ANTIBIOTICS》;20000831;第53卷(第8期);第844-847页 * |
| Bis-naphtho-γ-pyrones from Fungi and Their Bioactivities;Shiqiong Lu 等;《Molecules》;20140530;第19卷;第7169-7188页 * |
| Naphtho-Gamma-Pyrones Produced by Aspergillus tubingensis G131: New Source of Natural Nontoxic Antioxidants;Quentin Carboué 等;《Biomolecules》;20191224;第10卷(第29期);第1-15页 * |
| 共生曲霉菌D来源的聚酮类化合物及其生物合成初探;华熠;《中国优秀硕士学位论文全文数据库 (医药卫生科技辑)》;20200315(第3期);摘要 * |
| 海洋真菌Aspergillus niger XJJ-3 中萘并吡喃酮类化合物结构及生物活性;姜薇 等;《微生物学通报》;20180920;第45卷(第9期);第1897-1903页 * |
| 海藻内生真菌Aspergillus niger EN-13中的萘并吡喃酮类次生代谢产物;张翼 等;《万方数据知识服务平台》;全国海洋生物技术与海洋药物学术会议论文集;20071105;摘要，图1 * |
Also Published As
| Publication number | Publication date |
|---|---|
| CN113444644A (en) | 2021-09-28 |
| CN114848628B (en) | 2023-07-25 |
| CN114848628A (en) | 2022-08-05 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN103555607B (en) | A kind of endophyte H6 strain separation methods in silk tree blade and its application | |
| CN113444644B (en) | Application of naphtho-gamma-pyrone compounds in preparation of anti-helicobacter pylori medicines or prevention health-care products | |
| CN101126102A (en) | Marine actinomycetes for generating antineoplastic compound Norharmane | |
| CN116376722A (en) | Usnea longissima endophytic fungus YAFEF048 strain and preparation method and application thereof | |
| CN103724290A (en) | Cyclopeptide compound clavatustide A as well as producing strain, preparation method and application thereof | |
| Hao et al. | Isolation and identification of swainsonine-producing fungi found in locoweeds and their rhizosphere soil | |
| CN113234050B (en) | Antibacterial compound, preparation method and application thereof | |
| CN115806913A (en) | Streptomyces nojiri (Streptomyces nojiriensis) strain 9-13 and application thereof | |
| CN115536645A (en) | Compound Phonolide B, preparation method thereof and application thereof in antibacterial drugs | |
| CN113502230B (en) | Hericium erinaceus strain and culture method thereof, hericium erinaceus-ginseng bidirectional solid fermentation method and method for efficiently converting rare ginsenoside | |
| Kaushik et al. | Identification, optimization of culture conditions, and bioactive potential of Chinese caterpillar mushroom Ophiocordyceps sinensis (Ascomycetes) mycelium isolated from fruiting body | |
| JPH10503368A (en) | Antigenic animal cyclic tetrapeptide | |
| CN103641791B (en) | Cyclopeptide compound clavatustide B, and preparation method and application thereof | |
| CN103805543B (en) | A kind of bacterial strain and application thereof producing herbimycin | |
| CN108486011B (en) | Terphenyl compound, preparation method and application thereof | |
| US20100152467A1 (en) | Method for producing cercosporamide | |
| CN104711198A (en) | Penicillium ochrochloron and its application in preparation of brefeldin A | |
| KR100487703B1 (en) | Physiologically active substances tkr1785's, process for the preparation thereof, and microbe | |
| CN114703064B (en) | Method for extracting ganoderma lucidum by breaking cell walls of ganoderma lucidum and breaking cell walls of ganoderma lucidum by using trichoderma harzianum | |
| CN114350529B (en) | Licorice endophytic fungus for producing chalcone components and application thereof | |
| CN113278545B (en) | Streptomyces mutant and application thereof | |
| CN115466241B (en) | Compound with helicobacter pylori resisting activity and application thereof | |
| CN111909856B (en) | Preparation method and antibacterial application of ring-opened aromatic butenolide | |
| CN115287236B (en) | Preparation method and application of alkaloid compounds derived from insect symbiotic actinomycetes | |
| CN116004392B (en) | Antrodia camphorata symbiotic fungus YAFEF009 and separation method thereof |
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 |