CN117126105A - 一种吡啶甲酸类生物碱及其制备方法与应用 - Google Patents
一种吡啶甲酸类生物碱及其制备方法与应用 Download PDFInfo
- Publication number
- CN117126105A CN117126105A CN202311077059.6A CN202311077059A CN117126105A CN 117126105 A CN117126105 A CN 117126105A CN 202311077059 A CN202311077059 A CN 202311077059A CN 117126105 A CN117126105 A CN 117126105A
- Authority
- CN
- China
- Prior art keywords
- gastric cancer
- picolinic acid
- ethyl acetate
- compound
- cancer cells
- 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.)
- Pending
Links
- SIOXPEMLGUPBBT-UHFFFAOYSA-N picolinic acid Chemical class OC(=O)C1=CC=CC=N1 SIOXPEMLGUPBBT-UHFFFAOYSA-N 0.000 title claims abstract description 22
- 238000002360 preparation method Methods 0.000 title claims abstract description 11
- 208000005718 Stomach Neoplasms Diseases 0.000 claims abstract description 47
- 206010017758 gastric cancer Diseases 0.000 claims abstract description 46
- 201000011549 stomach cancer Diseases 0.000 claims abstract description 46
- -1 picolinic acid alkaloid compound Chemical class 0.000 claims abstract description 36
- 238000000855 fermentation Methods 0.000 claims abstract description 15
- 230000004151 fermentation Effects 0.000 claims abstract description 15
- 241000233866 Fungi Species 0.000 claims abstract description 11
- 239000003814 drug Substances 0.000 claims abstract description 9
- OKKJLVBELUTLKV-UHFFFAOYSA-N Methanol Chemical compound OC OKKJLVBELUTLKV-UHFFFAOYSA-N 0.000 claims description 24
- XEKOWRVHYACXOJ-UHFFFAOYSA-N Ethyl acetate Chemical compound CCOC(C)=O XEKOWRVHYACXOJ-UHFFFAOYSA-N 0.000 claims description 18
- 239000003208 petroleum Substances 0.000 claims description 16
- SRCZQMGIVIYBBJ-UHFFFAOYSA-N ethoxyethane;ethyl acetate Chemical compound CCOCC.CCOC(C)=O SRCZQMGIVIYBBJ-UHFFFAOYSA-N 0.000 claims description 14
- 241000234435 Lilium Species 0.000 claims description 9
- 238000001514 detection method Methods 0.000 claims description 9
- 239000007791 liquid phase Substances 0.000 claims description 9
- 238000000034 method Methods 0.000 claims description 9
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 9
- 239000000287 crude extract Substances 0.000 claims description 8
- 238000010828 elution Methods 0.000 claims description 8
- 241001157003 Penicillium brocae Species 0.000 claims description 6
- 239000003480 eluent Substances 0.000 claims description 6
- 230000014759 maintenance of location Effects 0.000 claims description 6
- 239000012071 phase Substances 0.000 claims description 6
- 244000005700 microbiome Species 0.000 claims description 4
- 150000003839 salts Chemical class 0.000 claims description 4
- 238000002791 soaking Methods 0.000 claims description 4
- 239000008194 pharmaceutical composition Substances 0.000 claims description 3
- 230000003213 activating effect Effects 0.000 claims description 2
- 239000012837 first-line chemotherapeutic agent Substances 0.000 claims description 2
- 238000010898 silica gel chromatography Methods 0.000 claims description 2
- 230000002538 fungal effect Effects 0.000 claims 1
- 150000001875 compounds Chemical class 0.000 abstract description 27
- 230000000694 effects Effects 0.000 abstract description 15
- GHASVSINZRGABV-UHFFFAOYSA-N Fluorouracil Chemical compound FC1=CNC(=O)NC1=O GHASVSINZRGABV-UHFFFAOYSA-N 0.000 abstract description 11
- 230000005764 inhibitory process Effects 0.000 abstract description 10
- 230000015572 biosynthetic process Effects 0.000 abstract description 7
- 229940079593 drug Drugs 0.000 abstract description 7
- 229960002949 fluorouracil Drugs 0.000 abstract description 7
- 229940044683 chemotherapy drug Drugs 0.000 abstract description 4
- 239000002246 antineoplastic agent Substances 0.000 abstract description 3
- 230000006907 apoptotic process Effects 0.000 abstract description 3
- 230000009545 invasion Effects 0.000 abstract description 3
- 238000011160 research Methods 0.000 abstract description 3
- 230000002195 synergetic effect Effects 0.000 abstract description 3
- 241001634096 Lilium martagon Species 0.000 abstract description 2
- 238000005516 engineering process Methods 0.000 abstract description 2
- 238000004519 manufacturing process Methods 0.000 abstract description 2
- 230000000813 microbial effect Effects 0.000 abstract description 2
- 239000001963 growth medium Substances 0.000 description 20
- 239000000243 solution Substances 0.000 description 15
- 238000005481 NMR spectroscopy Methods 0.000 description 13
- 238000011218 seed culture Methods 0.000 description 11
- 229930013930 alkaloid Natural products 0.000 description 10
- 230000003287 optical effect Effects 0.000 description 10
- 238000012258 culturing Methods 0.000 description 9
- 241000209094 Oryza Species 0.000 description 8
- 235000007164 Oryza sativa Nutrition 0.000 description 8
- 239000000047 product Substances 0.000 description 8
- 235000009566 rice Nutrition 0.000 description 8
- 239000000843 powder Substances 0.000 description 7
- 235000002639 sodium chloride Nutrition 0.000 description 6
- 238000001819 mass spectrum Methods 0.000 description 5
- 239000012528 membrane Substances 0.000 description 5
- 102000004169 proteins and genes Human genes 0.000 description 5
- 108090000623 proteins and genes Proteins 0.000 description 5
- 238000011282 treatment Methods 0.000 description 5
- RTZKZFJDLAIYFH-UHFFFAOYSA-N Diethyl ether Chemical compound CCOCC RTZKZFJDLAIYFH-UHFFFAOYSA-N 0.000 description 4
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 description 4
- 238000012512 characterization method Methods 0.000 description 4
- 239000013078 crystal Substances 0.000 description 4
- LOKCTEFSRHRXRJ-UHFFFAOYSA-I dipotassium trisodium dihydrogen phosphate hydrogen phosphate dichloride Chemical compound P(=O)(O)(O)[O-].[K+].P(=O)(O)([O-])[O-].[Na+].[Na+].[Cl-].[K+].[Cl-].[Na+] LOKCTEFSRHRXRJ-UHFFFAOYSA-I 0.000 description 4
- 238000002330 electrospray ionisation mass spectrometry Methods 0.000 description 4
- 238000000605 extraction Methods 0.000 description 4
- 238000004128 high performance liquid chromatography Methods 0.000 description 4
- 229910052739 hydrogen Inorganic materials 0.000 description 4
- 238000011534 incubation Methods 0.000 description 4
- 239000002609 medium Substances 0.000 description 4
- 239000002953 phosphate buffered saline Substances 0.000 description 4
- 238000004321 preservation Methods 0.000 description 4
- 230000035755 proliferation Effects 0.000 description 4
- 239000011780 sodium chloride Substances 0.000 description 4
- 238000001228 spectrum Methods 0.000 description 4
- 239000000126 substance Substances 0.000 description 4
- 230000004083 survival effect Effects 0.000 description 4
- 108091003079 Bovine Serum Albumin Proteins 0.000 description 3
- 102400000888 Cholecystokinin-8 Human genes 0.000 description 3
- 101800005151 Cholecystokinin-8 Proteins 0.000 description 3
- 238000004458 analytical method Methods 0.000 description 3
- 238000004364 calculation method Methods 0.000 description 3
- 229910052799 carbon Inorganic materials 0.000 description 3
- 239000003153 chemical reaction reagent Substances 0.000 description 3
- 238000010367 cloning Methods 0.000 description 3
- 231100000673 dose–response relationship Toxicity 0.000 description 3
- 238000002474 experimental method Methods 0.000 description 3
- 239000012091 fetal bovine serum Substances 0.000 description 3
- 239000012679 serum free medium Substances 0.000 description 3
- IZTQOLKUZKXIRV-YRVFCXMDSA-N sincalide Chemical compound C([C@@H](C(=O)N[C@@H](CCSC)C(=O)NCC(=O)N[C@@H](CC=1C2=CC=CC=C2NC=1)C(=O)N[C@@H](CCSC)C(=O)N[C@@H](CC(O)=O)C(=O)N[C@@H](CC=1C=CC=CC=1)C(N)=O)NC(=O)[C@@H](N)CC(O)=O)C1=CC=C(OS(O)(=O)=O)C=C1 IZTQOLKUZKXIRV-YRVFCXMDSA-N 0.000 description 3
- 230000001954 sterilising effect Effects 0.000 description 3
- 239000006228 supernatant Substances 0.000 description 3
- LDVVTQMJQSCDMK-UHFFFAOYSA-N 1,3-dihydroxypropan-2-yl formate Chemical compound OCC(CO)OC=O LDVVTQMJQSCDMK-UHFFFAOYSA-N 0.000 description 2
- OKTJSMMVPCPJKN-UHFFFAOYSA-N Carbon Chemical compound [C] OKTJSMMVPCPJKN-UHFFFAOYSA-N 0.000 description 2
- 241000196324 Embryophyta Species 0.000 description 2
- UFHFLCQGNIYNRP-UHFFFAOYSA-N Hydrogen Chemical compound [H][H] UFHFLCQGNIYNRP-UHFFFAOYSA-N 0.000 description 2
- 206010027476 Metastases Diseases 0.000 description 2
- 101710135898 Myc proto-oncogene protein Proteins 0.000 description 2
- 102100038895 Myc proto-oncogene protein Human genes 0.000 description 2
- 229930040373 Paraformaldehyde Natural products 0.000 description 2
- 108010064218 Poly (ADP-Ribose) Polymerase-1 Proteins 0.000 description 2
- 102100023712 Poly [ADP-ribose] polymerase 1 Human genes 0.000 description 2
- 102100040678 Programmed cell death protein 1 Human genes 0.000 description 2
- 101710089372 Programmed cell death protein 1 Proteins 0.000 description 2
- VYPSYNLAJGMNEJ-UHFFFAOYSA-N Silicium dioxide Chemical compound O=[Si]=O VYPSYNLAJGMNEJ-UHFFFAOYSA-N 0.000 description 2
- 101710150448 Transcriptional regulator Myc Proteins 0.000 description 2
- 102000016549 Vascular Endothelial Growth Factor Receptor-2 Human genes 0.000 description 2
- 108010053099 Vascular Endothelial Growth Factor Receptor-2 Proteins 0.000 description 2
- 230000009471 action Effects 0.000 description 2
- 230000000259 anti-tumor effect Effects 0.000 description 2
- 230000003833 cell viability Effects 0.000 description 2
- 238000013375 chromatographic separation Methods 0.000 description 2
- 230000007547 defect Effects 0.000 description 2
- 230000002950 deficient Effects 0.000 description 2
- 238000007865 diluting Methods 0.000 description 2
- 238000002114 high-resolution electrospray ionisation mass spectrometry Methods 0.000 description 2
- 239000001257 hydrogen Substances 0.000 description 2
- 230000002401 inhibitory effect Effects 0.000 description 2
- 239000000463 material Substances 0.000 description 2
- 230000009401 metastasis Effects 0.000 description 2
- 239000000203 mixture Substances 0.000 description 2
- 230000005311 nuclear magnetism Effects 0.000 description 2
- 229920002866 paraformaldehyde Polymers 0.000 description 2
- 238000005191 phase separation Methods 0.000 description 2
- 239000011148 porous material Substances 0.000 description 2
- 238000000746 purification Methods 0.000 description 2
- 239000000741 silica gel Substances 0.000 description 2
- 229910002027 silica gel Inorganic materials 0.000 description 2
- 238000002415 sodium dodecyl sulfate polyacrylamide gel electrophoresis Methods 0.000 description 2
- UCSJYZPVAKXKNQ-HZYVHMACSA-N streptomycin Chemical compound CN[C@H]1[C@H](O)[C@@H](O)[C@H](CO)O[C@H]1O[C@@H]1[C@](C=O)(O)[C@H](C)O[C@H]1O[C@@H]1[C@@H](NC(N)=N)[C@H](O)[C@@H](NC(N)=N)[C@H](O)[C@H]1O UCSJYZPVAKXKNQ-HZYVHMACSA-N 0.000 description 2
- 230000008685 targeting Effects 0.000 description 2
- 229960000575 trastuzumab Drugs 0.000 description 2
- 238000005406 washing Methods 0.000 description 2
- 238000001262 western blot Methods 0.000 description 2
- QAPSNMNOIOSXSQ-YNEHKIRRSA-N 1-[(2r,4s,5r)-4-[tert-butyl(dimethyl)silyl]oxy-5-(hydroxymethyl)oxolan-2-yl]-5-methylpyrimidine-2,4-dione Chemical compound O=C1NC(=O)C(C)=CN1[C@@H]1O[C@H](CO)[C@@H](O[Si](C)(C)C(C)(C)C)C1 QAPSNMNOIOSXSQ-YNEHKIRRSA-N 0.000 description 1
- 238000000035 BCA protein assay Methods 0.000 description 1
- 238000007808 Cell invasion assay Methods 0.000 description 1
- 229920000742 Cotton Polymers 0.000 description 1
- 102000004190 Enzymes Human genes 0.000 description 1
- 108090000790 Enzymes Proteins 0.000 description 1
- 206010066896 HER-2 positive gastric cancer Diseases 0.000 description 1
- 101710088172 HTH-type transcriptional regulator RipA Proteins 0.000 description 1
- 101001012157 Homo sapiens Receptor tyrosine-protein kinase erbB-2 Proteins 0.000 description 1
- 101000911513 Homo sapiens Uncharacterized protein FAM215A Proteins 0.000 description 1
- 206010028980 Neoplasm Diseases 0.000 description 1
- 239000000020 Nitrocellulose Substances 0.000 description 1
- 241000283973 Oryctolagus cuniculus Species 0.000 description 1
- 239000002033 PVDF binder Substances 0.000 description 1
- 229930182555 Penicillin Natural products 0.000 description 1
- JGSARLDLIJGVTE-MBNYWOFBSA-N Penicillin G Chemical compound N([C@H]1[C@H]2SC([C@@H](N2C1=O)C(O)=O)(C)C)C(=O)CC1=CC=CC=C1 JGSARLDLIJGVTE-MBNYWOFBSA-N 0.000 description 1
- 102100030086 Receptor tyrosine-protein kinase erbB-2 Human genes 0.000 description 1
- 102100026728 Uncharacterized protein FAM215A Human genes 0.000 description 1
- 238000002835 absorbance Methods 0.000 description 1
- 230000004913 activation Effects 0.000 description 1
- 238000011226 adjuvant chemotherapy Methods 0.000 description 1
- 150000003797 alkaloid derivatives Chemical class 0.000 description 1
- 230000003698 anagen phase Effects 0.000 description 1
- 230000001093 anti-cancer Effects 0.000 description 1
- 238000013459 approach Methods 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 230000003115 biocidal effect Effects 0.000 description 1
- 230000004071 biological effect Effects 0.000 description 1
- 239000000872 buffer Substances 0.000 description 1
- 201000011510 cancer Diseases 0.000 description 1
- 239000003560 cancer drug Substances 0.000 description 1
- 238000004113 cell culture Methods 0.000 description 1
- 239000013592 cell lysate Substances 0.000 description 1
- 238000001816 cooling Methods 0.000 description 1
- 238000004925 denaturation Methods 0.000 description 1
- 230000036425 denaturation Effects 0.000 description 1
- 238000003745 diagnosis Methods 0.000 description 1
- 230000002900 effect on cell Effects 0.000 description 1
- 238000011354 first-line chemotherapy Methods 0.000 description 1
- 239000000499 gel Substances 0.000 description 1
- 238000009650 gentamicin protection assay Methods 0.000 description 1
- 230000007365 immunoregulation Effects 0.000 description 1
- 238000000338 in vitro Methods 0.000 description 1
- 239000003112 inhibitor Substances 0.000 description 1
- 239000006166 lysate Substances 0.000 description 1
- 238000004949 mass spectrometry Methods 0.000 description 1
- 230000002503 metabolic effect Effects 0.000 description 1
- 238000002156 mixing Methods 0.000 description 1
- 238000012986 modification Methods 0.000 description 1
- 230000004048 modification Effects 0.000 description 1
- 238000011228 multimodal treatment Methods 0.000 description 1
- 229920001220 nitrocellulos Polymers 0.000 description 1
- 229960003301 nivolumab Drugs 0.000 description 1
- 238000000655 nuclear magnetic resonance spectrum Methods 0.000 description 1
- 229940049954 penicillin Drugs 0.000 description 1
- 125000001997 phenyl group Chemical group [H]C1=C([H])C([H])=C(*)C([H])=C1[H] 0.000 description 1
- 230000001766 physiological effect Effects 0.000 description 1
- 229920002401 polyacrylamide Polymers 0.000 description 1
- 229920002981 polyvinylidene fluoride Polymers 0.000 description 1
- 238000010814 radioimmunoprecipitation assay Methods 0.000 description 1
- 229960002633 ramucirumab Drugs 0.000 description 1
- 238000002271 resection Methods 0.000 description 1
- 235000019991 rice wine Nutrition 0.000 description 1
- 238000007790 scraping Methods 0.000 description 1
- 238000012216 screening Methods 0.000 description 1
- 238000007789 sealing Methods 0.000 description 1
- 229930000044 secondary metabolite Natural products 0.000 description 1
- 238000012163 sequencing technique Methods 0.000 description 1
- 235000020183 skimmed milk Nutrition 0.000 description 1
- 150000003384 small molecules Chemical class 0.000 description 1
- 229960005322 streptomycin Drugs 0.000 description 1
- 238000006467 substitution reaction Methods 0.000 description 1
- 230000007761 synergistic anti-cancer Effects 0.000 description 1
- 238000002626 targeted therapy Methods 0.000 description 1
- 238000012360 testing method Methods 0.000 description 1
- 229940126585 therapeutic drug Drugs 0.000 description 1
- 238000011269 treatment regimen Methods 0.000 description 1
Classifications
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07D—HETEROCYCLIC COMPOUNDS
- C07D213/00—Heterocyclic compounds containing six-membered rings, not condensed with other rings, with one nitrogen atom as the only ring hetero atom and three or more double bonds between ring members or between ring members and non-ring members
- C07D213/02—Heterocyclic compounds containing six-membered rings, not condensed with other rings, with one nitrogen atom as the only ring hetero atom and three or more double bonds between ring members or between ring members and non-ring members having three double bonds between ring members or between ring members and non-ring members
- C07D213/04—Heterocyclic compounds containing six-membered rings, not condensed with other rings, with one nitrogen atom as the only ring hetero atom and three or more double bonds between ring members or between ring members and non-ring members having three double bonds between ring members or between ring members and non-ring members having no bond between the ring nitrogen atom and a non-ring member or having only hydrogen or carbon atoms directly attached to the ring nitrogen atom
- C07D213/60—Heterocyclic compounds containing six-membered rings, not condensed with other rings, with one nitrogen atom as the only ring hetero atom and three or more double bonds between ring members or between ring members and non-ring members having three double bonds between ring members or between ring members and non-ring members having no bond between the ring nitrogen atom and a non-ring member or having only hydrogen or carbon atoms directly attached to the ring nitrogen atom with hetero atoms or with carbon atoms having three bonds to hetero atoms with at the most one bond to halogen, e.g. ester or nitrile radicals, directly attached to ring carbon atoms
- C07D213/78—Carbon atoms having three bonds to hetero atoms, with at the most one bond to halogen, e.g. ester or nitrile radicals
- C07D213/79—Acids; Esters
-
- 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
- A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
- A61K31/435—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with one nitrogen as the only ring hetero atom
- A61K31/44—Non condensed pyridines; Hydrogenated derivatives thereof
- A61K31/4427—Non condensed pyridines; Hydrogenated derivatives thereof containing further heterocyclic ring systems
- A61K31/444—Non condensed pyridines; Hydrogenated derivatives thereof containing further heterocyclic ring systems containing a six-membered ring with nitrogen as a ring heteroatom, e.g. amrinone
-
- 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
- A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
- A61K31/495—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with two or more nitrogen atoms as the only ring heteroatoms, e.g. piperazine or tetrazines
- A61K31/505—Pyrimidines; Hydrogenated pyrimidines, e.g. trimethoprim
- A61K31/513—Pyrimidines; Hydrogenated pyrimidines, e.g. trimethoprim having oxo groups directly attached to the heterocyclic ring, e.g. cytosine
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P35/00—Antineoplastic agents
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07D—HETEROCYCLIC COMPOUNDS
- C07D213/00—Heterocyclic compounds containing six-membered rings, not condensed with other rings, with one nitrogen atom as the only ring hetero atom and three or more double bonds between ring members or between ring members and non-ring members
- C07D213/02—Heterocyclic compounds containing six-membered rings, not condensed with other rings, with one nitrogen atom as the only ring hetero atom and three or more double bonds between ring members or between ring members and non-ring members having three double bonds between ring members or between ring members and non-ring members
- C07D213/04—Heterocyclic compounds containing six-membered rings, not condensed with other rings, with one nitrogen atom as the only ring hetero atom and three or more double bonds between ring members or between ring members and non-ring members having three double bonds between ring members or between ring members and non-ring members having no bond between the ring nitrogen atom and a non-ring member or having only hydrogen or carbon atoms directly attached to the ring nitrogen atom
- C07D213/60—Heterocyclic compounds containing six-membered rings, not condensed with other rings, with one nitrogen atom as the only ring hetero atom and three or more double bonds between ring members or between ring members and non-ring members having three double bonds between ring members or between ring members and non-ring members having no bond between the ring nitrogen atom and a non-ring member or having only hydrogen or carbon atoms directly attached to the ring nitrogen atom with hetero atoms or with carbon atoms having three bonds to hetero atoms with at the most one bond to halogen, e.g. ester or nitrile radicals, directly attached to ring carbon atoms
- C07D213/78—Carbon atoms having three bonds to hetero atoms, with at the most one bond to halogen, e.g. ester or nitrile radicals
- C07D213/79—Acids; Esters
- C07D213/803—Processes of preparation
-
- 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
- C12P17/10—Nitrogen as only ring hetero atom
- C12P17/12—Nitrogen as only ring hetero atom containing a six-membered hetero ring
-
- 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
-
- 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
- Chemical & Material Sciences (AREA)
- Organic Chemistry (AREA)
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- General Health & Medical Sciences (AREA)
- Veterinary Medicine (AREA)
- Medicinal Chemistry (AREA)
- Public Health (AREA)
- Pharmacology & Pharmacy (AREA)
- Animal Behavior & Ethology (AREA)
- Zoology (AREA)
- Wood Science & Technology (AREA)
- General Chemical & Material Sciences (AREA)
- Epidemiology (AREA)
- Engineering & Computer Science (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Microbiology (AREA)
- Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
- Biotechnology (AREA)
- Biochemistry (AREA)
- Bioinformatics & Cheminformatics (AREA)
- General Engineering & Computer Science (AREA)
- Genetics & Genomics (AREA)
- Nitrogen Condensed Heterocyclic Rings (AREA)
Abstract
本发明属于生物医药技术领域,具体涉及一种吡啶甲酸类生物碱及其制备方法与应用。该化合物可以利用南海海百合来源真菌通过现代微生物发酵工程技术得到,易实现大规模产业化生产。并且研究发现,所述吡啶甲酸类生物碱化合物对胃癌细胞具有显著的抑制作用,可以显著抑制胃癌细胞的活性和克隆形成,促进胃癌细胞的凋亡,减少胃癌细胞的侵袭;当将吡啶甲酸类生物碱化合物与临床一线化疗药如5‑氟尿嘧啶联合作用于胃癌细胞时,细胞抑制效果还得到了显著的提升,具有协同增效的抗胃癌效果。
Description
技术领域
本发明属于生物医药技术领域。更具体地,涉及一种吡啶甲酸类生物碱及其制备方法与应用。
背景技术
胃癌是世界上最常见的恶性肿瘤之一,在所有肿瘤中的发病率排名第五位,死亡率排名第四位。并且,大约60%的胃癌患者在初诊时被诊断为伴有局部或远处转移,这些患者的5年总生存率分别仅为30%和5%左右。手术切除和围术期辅助化疗提高了局部进展期胃癌的生存率,但对于伴有局部或远处转移的患者治疗策略仍然有限。以小分子抑制剂和单克隆抗体为形式的靶向治疗已成为胃癌多模式治疗的重要方法,如曲妥珠单抗作为HER2单克隆抗体,已经被批准用于晚期HER2阳性胃癌的一线治疗(比如中国专利申请CN104045714A公开了曲妥珠单抗突变体IgG及其应用)。此外,雷莫芦单抗(一种靶向血管内皮生长因子受体2(VEGFR-2)的抗体)和纳武单抗或帕博利珠单抗(靶向程序性细胞死亡蛋白1(PD-1)的抗体)作为二线和三线治疗也显著提高了胃癌患者的无进展生存期和总生存率。但在临床治疗中,针对胃癌患者的治疗药物仍相对匮乏,因此,迫切需要探索更多新的有效的针对胃癌患者的药物,以提高临床治疗的有效性。
发明内容
本发明要解决的技术问题是克服现有胃癌患者治疗药物相对匮乏的缺陷和不足,提供一种对胃癌具有显著抑制作用的吡啶甲酸类生物碱。
本发明的目的是提供所述吡啶甲酸类生物碱的制备方法。
本发明另一目的是提供所述吡啶甲酸类生物碱在制备抗胃癌药物中的应用。
本发明还有一个目的是提供一种抗胃癌的药物组合物。
本发明上述目的通过以下技术方案实现:
一种吡啶甲酸类生物碱,具有式(I)、(II)结构:
生物碱是广泛分布于植物体内的一类大多呈碱性的含氮有机化合物,绝大多数分布在高等植物体内,并且具有复杂的结构和显著的生物活性,不少生物碱还被发现具有抗肿瘤活性。海洋生物在特殊环境之中,已发展出独特的代谢方式,很多文献已证明海洋生物来源的真菌能够产生结构新颖、生理活性显著的各类次级代谢产物,有抗菌、抗肿瘤、免疫调节、酶抑制等多种药用价值。本发明所提供的吡啶甲酸类生物碱就是从海洋微生物的发酵产物中提取、分离、纯化得到的。
因此,本发明还提供了所述吡啶甲酸类生物碱的制备方法,所述吡啶甲酸类生物碱由南海海百合来源真菌发酵产物提取、分离、纯化得到。
进一步地,所述南海海百合来源真菌命名为Penicillium brocae SYSU-CJ-17,保藏在广东省微生物菌种保藏中心,保藏日期是2023年05月26日,保藏号是GDMCC No:63444。
更进一步地,所述制备方法具体包括以下步骤:
S1、将南海海百合来源真菌活化、扩大培养,得到发酵物;
S2、将步骤S1所得发酵物用乙酸乙酯浸泡提取,提取液浓缩得到粗浸膏;
S3、将步骤S2所得粗浸膏进行硅胶柱层析,以石油醚-乙酸乙酯作为洗脱液进行梯度洗脱,收集石油醚-乙酸乙酯体积比为1:1的流份;
S4、将步骤S3所得石油醚-乙酸乙酯体积比为1:1的流份高效液相进行分离纯化,得到吡啶甲酸类生物碱。
进一步地,步骤S3中,所述梯度洗脱为以石油醚-乙酸乙酯作为洗脱液,按照石油醚-乙酸乙酯体积比5:1、4:1、3:1、2:1、1:1、2:3、1:4、0:1依次进行梯度洗脱。
更进一步地,制备吡啶甲酸类生物碱化合物(I)时,步骤S4中的高效液相条件为流动相为50-70%甲醇/水(具体为50-70%甲醇和30~50%的水,两者加起来为100%),流速为2-3mL/min,检测波长为230nm或275nm,保留时间为10-20min。
进一步地,制备吡啶甲酸类生物碱化合物(II)时,步骤S4中的高效液相条件为流动相为50-70%甲醇/水(具体为50-70%甲醇和30~50%的水,两者加起来为100%),流速为2-3mL/min,检测波长为230nm或275nm,保留时间为5-15min。
优选地,所述吡啶甲酸类生物碱化合物采用本发明方法可以通过出峰先后顺序来区分;其中,吡啶甲酸类生物碱化合物(II)先出峰。
优选地,高效液相所采用的色谱柱为半制备的苯基柱;具体可以为半制备柱Ultimate XB-Phenyl,10×250mm,5μm;仪器为Essentia LC-16。
更进一步地,步骤S1中,所述活化、扩大培养为先制备种子培养液,再进行发酵培养。
具体的,种子培养液的制备采用的培养基为含有25-35g/L海盐和20-30g/LPDB培养基粉末的种子培养基;培养条件为摇床转速100~150rpm,25-28℃恒温培养3~5天。
发酵培养采用的培养基为大米培养基,其中,所述大米培养基为40~50g大米,50mL 2~3%的盐水,置于培养瓶混匀后密封,经高温灭菌锅121℃(0.1MPa)高温灭菌25min冷却至室温,放置2天,即得;优选地,所述发酵培养的条件为25-28℃静置培养28~32天。
研究发现,本发明所提供的吡啶甲酸类生物碱或其药学上可接受的盐对胃癌具有显著的抑制作用,因此,本发明还要求保护所述吡啶甲酸类生物碱在制备抗胃癌药物中的应用。
进一步地,所述胃癌为MGC803、MKN45、HGC27、AGS、SNU1或KATO3胃癌细胞系造成的。
另外的,本发明还提供了一种抗胃癌的药物组合物,所述组合物包括所述吡啶甲酸类生物碱或其药学上可接受的盐和临床一线化疗药物;优选的,所述临床一线化疗药物可以为5-氟尿嘧啶。实验证明,所述吡啶甲酸类生物碱和5-氟尿嘧啶(5-Fu)联合应用作用于胃癌细胞时,能显著抑制胃癌细胞的细胞活力以及胃癌细胞的克隆形成,具有显著的协同抗癌作用。
本发明具有以下有益效果:
本发明提供了一种吡啶甲酸类生物碱化合物,该化合物可以利用南海海百合来源真菌通过现代微生物发酵工程技术得到,易实现大规模产业化生产。并且研究发现,所述吡啶甲酸类生物碱化合物对胃癌细胞具有显著的抑制作用,可以显著抑制胃癌细胞的活性和克隆形成,促进胃癌细胞的凋亡,减少胃癌细胞的侵袭;当将吡啶甲酸类生物碱化合物与5-氟尿嘧啶联合作用于胃癌细胞时,细胞抑制效果还得到了显著的提升,具有协同增效的抗胃癌效果。
附图说明
图1为本发明实施例1所得吡啶甲酸类生物碱化合物(I)的核磁共振氢谱图。
图2为本发明实施例1所得吡啶甲酸类生物碱化合物(I)的核磁共振碳谱图。
图3为本发明实施例1所得吡啶甲酸类生物碱化合物(I)的HRESIMS质谱图。
图4为本发明实施例2所得吡啶甲酸类生物碱化合物(II)的核磁共振氢谱图。
图5为本发明实施例2所得吡啶甲酸类生物碱化合物(II)的核磁共振碳谱图。
图6为本发明实施例2所得吡啶甲酸类生物碱化合物(II)的HRESIMS质谱图。
图7为本发明所得吡啶甲酸类生物碱化合物(I)或(II)对各种胃癌细胞系(MGC803、MKN45、HGC27、AGS、SNU1、KATO3)的抑制活性筛选数据统计图。
图8为不同浓度吡啶甲酸类生物碱化合物(II)对各种胃癌细胞系的抑制活性数据统计图。
图9为不同浓度吡啶甲酸类生物碱化合物(II)胃癌细胞的克隆形成显微镜图和数据统计图。
图10为不同浓度吡啶甲酸类生物碱化合物(II)对不同胃癌细胞系(MGC803、MKN45)凋亡相关蛋白(PARP-1、cPARP-1、c-Caspase7)和增殖相关蛋白(c-Myc、cyclinD)作用的WB图。
图11为不同浓度吡啶甲酸类生物碱化合物(II)胃癌细胞的侵袭显微镜图和数据统计图。
图12为吡啶甲酸类生物碱化合物(II)与5-Fu各自单独作用或联用对胃癌细胞的细胞活力抑制的数据统计图。
图13为吡啶甲酸类生物碱化合物(II)与5-Fu各自单独作用或联用对胃癌细胞的克隆形成的显微镜图。
具体实施方式
以下结合说明书附图和具体实施例来进一步说明本发明,但实施例并不对本发明做任何形式的限定。除非特别说明,本发明采用的试剂、方法和设备为本技术领域常规试剂、方法和设备。
本发明实施例中所采用的真菌为海洋真菌,该海洋真菌为一株来自中国南海海百合分离得到的真菌,经过分子生物学鉴定,其分类为Penicillium brocae,真菌命名为Penicillium brocae SYSU-CJ-17,保藏在广东省微生物菌种保藏中心,保藏日期是2023年05月26日,保藏号是GDMCC No:63444。
种子培养基:按照每升水中加入30g海盐和24g PDB培养基粉末进行配制,平均分配到4个1L锥形瓶内,经高温灭菌锅121℃(0.1MPa)条件灭菌25min,冷却至室温,静置24小时,即得。
大米培养基:50g大米,50mL3%的盐水,置于培养瓶混匀后密封,经高温灭菌锅121℃(0.1MPa)高温灭菌25min冷却至室温,放置2天,即得。
除非特别说明,以下实施例所用试剂和材料均为市购。
实施例1吡啶甲酸类生物碱化合物(I)的提取与表征
1、吡啶甲酸类生物碱化合物(I)的提取具体包括以下步骤:
S1、种子培养液的获得:将南海海百合来源真菌Penicillium brocae SYSU-CJ-17接入种子培养基中,将接种后的锥形瓶放置在摇床上,摇床转速100~150rpm,25℃恒温培养72小时,获得种子培养液。
S2、发酵培养:挑选瓶内培养基无染菌现象的进行接种,每瓶接种10mL菌种(步骤S1所得种子培养液),共接种200瓶,静置培养30天得发酵物。
S3、提取分离:发酵培养后,将S2所得发酵物用乙酸乙酯浸泡提取,减压浓缩提取液得粗浸膏;分别以不同体积比的石油醚-乙酸乙酯作为洗脱剂,依次用体积比为5:1、4:1、3:1、2:1、1:1、2:3、1:4、0:1的石油醚-乙酸乙酯进行梯度洗脱,对粗浸膏采用硅胶柱进行层析分离,分别得到不同极性组分。
S4、高效液相分离纯化:将步骤S3中所得石油醚-乙酸乙酯体积比为1:1的洗脱部分通过高效液相色谱分离纯化,所述高效液相色谱的条件为:流动相:60% MeOH-40%H2O;流速:3mL/min,色谱柱:半制备柱Ultimate XB-Phenyl,10×250mm,5μm;仪器Essentia LC-16,检测波长为230nm或275nm,在保留时间12min处得到白色粉末。
2、吡啶甲酸类生物碱化合物(I)的表征:
对所得白色粉末进行核磁共振和质谱检测,所得谱图如图1~3所示,经分析检测,该化合物结构的理化性质数据如下:
UV(MeOH)λmax(logε)229.6(4.08),269.4(3.78);
IR(neat)νmax 3367cm-1,2917cm-1,2844cm-1,1720cm-1,1574cm-1,1431cm-1,1389cm-1,1312cm-1,1254cm-1,1216cm-1,1154cm-1,1119cm-1,1077cm-1,1016cm-1,869,796cm-1,708cm-1,630cm-1;
HR-ESIMS m/s 515.2752[M+H]+(calcd for C28H39N2O7,515.2752),详细信息如表1所示。
表1吡啶甲酸类生物碱化合物(I)的HR-ESIMS信息
NMR核磁的数据:
13C NMR(150MHz,CDCl3)δC 171.11C,165.92C,165.06C,150.12CH,149.75CH,145.58C,144.93C,142.96C,142.35C,137.23CH,136.80CH,125.46CH,125.04CH,67.85CH,67.13CH2,65.08CH2,52.88CH3,33.15CH2,33.12CH2,30.92CH2,30.90CH2,29.52CH2,29.40CH2,29.17CH2,29.14CH2,20.93CH2.
1H NMR(600MHz,CDCl3)δH 8.54s,8.53s,8.06d,8.04d,7.65dd,7.62dd,4.47dd,4.41dd,4.29-4.25m,4.22d,3.98s,2.67td,2.07s,1.61q,1.33-1.20m。
从质谱和核磁共振的结果分析可确定化合物式(Ι)的分子式为C28H38N2O7。
从平面结构发现化合物(I)的一个手性中心C-19,由于该化合物具有一条柔性的长链,无法通过ECD计算得到准确的构型。但这种具有单甘油片段且只有一个手性中心的化合物手性可以通过旋光来判断,若旋光值若为负值,则该手性中心为S构型,若为正值则是R构型。
因此,测定白色粉末的旋光,得到该化合物的旋光数据为 可判断该化合物(I)的构型为19R。
综上所述,所得白色粉末为吡啶甲酸类生物碱化合物(I),具体结构式如下:
实施例2吡啶甲酸类生物碱化合物(II)的提取与表征
1、吡啶甲酸类生物碱化合物(II)的提取具体包括以下步骤:
S1、种子培养液的获得:将南海海百合来源真菌Penicillium brocae SYSU-CJ-17接入种子培养基中,将接种后的锥形瓶放置在摇床上,摇床转速100~150rpm,25℃恒温培养72小时,获得种子培养液。
S2、发酵培养:挑选瓶内培养基无染菌现象的进行接种,每瓶接种10mL菌种(步骤S1所得种子培养液),共接种200瓶,静置培养30天得发酵物。
S3、提取分离:发酵培养后,将S2所得发酵物用乙酸乙酯浸泡提取,减压浓缩提取液得粗浸膏;分别以不同体积比的石油醚-乙酸乙酯作为洗脱剂,依次用体积比为5:1、4:1、3:1、2:1、1:1、2:3、1:4、0:1的石油醚-乙酸乙酯进行梯度洗脱,对粗浸膏采用硅胶柱进行层析分离,分别得到不同极性组分。
S4、高效液相分离纯化:将步骤S3中所得石油醚-乙酸乙酯体积比为1:1的洗脱部分通过高效液相色谱分离纯化,所述高效液相色谱的条件为:流动相:60% MeOH-40%H2O;流速:3mL/min,色谱柱:半制备柱Ultimate XB-Phenyl,10×250mm,5μm;仪器Essentia LC-16,检测波长为230nm或275nm,在保留时间8.5min处得到淡黄色无定形物。
2、吡啶甲酸类生物碱化合物(II)的表征:
对所得淡黄色无定形物进行核磁共振和质谱检测,所得谱图如图4~6所示,经分析检测,该化合物结构的理化性质数据如下:
UV(MeOH)λmax(logε)229.8(4.07),268.7(3.78)nm;
IR(neat)νmax 3352cm-1,2913cm-1,2852cm-1,1704cm-1,1655cm-1,1570cm-1,1428cm-1,1389cm-1,1312cm-1,1246cm-1,1204cm-1,1116cm-1,1023cm-1,862cm-1,773cm-1,696cm-1,626cm-1;
HR-ESIMS m/s 473.2650[M+H]+(calcd for C26H37N2O6,473.2646),详细信息如表2所示。
表2吡啶甲酸类生物碱化合物(II)的HR-ESIMS信息
NMR核磁的数据:
13C NMR(150MHz,CD3OD)δC 166.57C,165.95C,150.71CH,150.67CH,146.27C,146.24C,144.54C,144.40C,138.85CH,138.75CH,126.42CH,126.18CH,71.13CH,68.15CH2,63.88CH2,53.10CH3,33.76CH2,31.97CH2,31.95CH2,30.52CH2,30.38CH2,30.17CH2.
1H NMR(600MHz,CD3OD)δH 8.51dd,8.12d,8.06d,7.83ddd,4.46dd 4.33dd,4.00dt,3.95s,3.65d,2.73q,1.68-1.61m,1.35-1.25m。
从质谱和核磁共振的结果分析可确定化合物式(ΙI)的分子式为C26H36N2O6。
从平面结构发现化合物(II)的一个手性中心C-19,由于该化合物具有一条柔性的长链,无法通过ECD计算得到准确的构型。但这种具有单甘油片段且只有一个手性中心的化合物手性可以通过旋光来判断,若旋光值若为负值,则该手性中心为S构型,若为正值则是R构型。
因此,测定所得淡黄色无定形物的旋光,得到该化合物的旋光数据为 可以判断该化合物的手性为19S。
综上所述,所得淡黄色无定形物为吡啶甲酸类生物碱化合物(II),具体结构式如下:
实施例3化合物的抗癌活性测试
1、细胞培养
本实施例中所用胃癌细胞系(MGC803、MKN45、HGC27、AGS、SNU1和KATO3)培养于含有10%胎牛血清(FBS)和1%青霉素/链霉素的RPIM-1640培养基(以下简称为培养基)中,细胞放置于含有5% CO2,温度为37℃的细胞培养箱中培养。
2、CCK8法测定细胞活力
将500个细胞均匀稀释于100μL培养基,接种于96孔板中,细胞培养24h后,将化合物(I)或化合物(II)稀释成不同浓度,并以50μL的培养基添加到每孔中;连续4天细胞孵育后加入CCK8试剂,96孔板在含5% CO2,37℃的培养箱中继续孵育1~2小时;根据CCK8试剂盒的说明在450nm处测量吸光度。最终结果显示为百分比,采用GraphPad Prism 7软件计算体外IC50值。其抑制率计算公式为[OD(加药组)-OD(对照组)]/[OD(对照组)-OD(空白组)]×100%。
结果参见图7-8,由图可见化合物(I)或(II)可以剂量依赖性的抑制胃癌细胞的增殖活性。
3、平板克隆实验-化合物(II)单独作用
将对数生长期的细胞以每孔500个均匀接种于六孔板中;培养细胞24h后加入不同浓度的药物,继续将六孔板置于37℃培养箱中孵育,每组设置3个复孔;每三天更换一次新鲜培养基,并再次加入不同浓度的化合物(II);培养10~12天后,用磷酸盐缓冲盐水(PBS)洗涤细胞,用4%多聚甲醛固定细胞15分钟。最后,用结晶紫溶液避光染色20分钟,用PBS洗涤结晶紫溶液后进行细胞集落计数,实验重复3次。
结果参见图9,由图可见化合物(II)可以通过抑制胃癌细胞克隆的形成抑制其增殖。
4、蛋白免疫印迹法(Western Blot)
用药物处理细胞48h后,收集细胞。收集的细胞用细胞裂解液RIPA处理约15分钟,后将裂解液在15000rpm,4℃条件下离心15min。收集并转移上清液,通过BCA蛋白测定法定量蛋白质浓度。随后加入5×loading buffer,并将蛋白质上清液煮沸变性。取适量蛋白质上清液加入6%~15%的聚丙烯酰胺凝胶中,经十二烷基硫酸钠-聚丙烯酰胺凝胶电泳(SDS-PAGE)分离样品约100min,后转移到硝酸纤维素薄膜(PVDF膜)上。用脱脂奶粉在室温下封闭膜1h,然后用TBS-T溶液洗3次,用相应的一抗在4℃下孵育过夜。第二天用兔二抗室温孵育1h。最后,利用显影仪对膜上的蛋白质条带进行可视化分析。
结果参见图10,由图可见化合物(II)可以剂量依赖性的上调凋亡蛋白PARP-1及c-caspase-7,并且抑制增殖蛋白c-myc及cyclinD的表达。
5、Transwell小室侵袭实验
细胞侵袭检测使用24孔板进行,加入无血清培养基,将小室的膜浸润在无血清培养基中,在37℃的培养箱中维持1h或过夜,以活化小室;将六孔板中不同浓度药物处理48h后的细胞进行消化和计数,将含有2×104个细胞的300μL无血清培养基均匀加到Transwell小室的上室中,下室加入500μL含有10%胎牛血清的培养基。将小室放置在含5%CO2,温度为37℃的培养箱中过夜培养;孵育24h后,用4%多聚甲醛固定细胞15min,结晶紫避光染色30min;用PBS轻轻冲洗掉结晶紫染液,用棉签轻轻刮掉小室上层的细胞;待小室干燥后在显微镜下拍照,进行计数统计。
结果参见图11,由图可见化合物(II)可以剂量依赖性的抑制胃癌细胞的侵袭能力。
6、平板克隆实验-化合物(II)与5-氟尿嘧啶(5-Fu)联合作用
参考第3点平板克隆实验测试化合物(II)与5-氟尿嘧啶(5-Fu)联合对细胞的影响。
结果参见图12-13,由图可见,化合物(II)(图中Pen-I)协同化疗药物可有效增强临床一线化疗药物5-氟尿嘧啶(5-Fu)对两种胃癌细胞系的细胞克隆形成的显著的抑制效果,两者之间具有较好的协同增效作用。
上述实施例为本发明较佳的实施方式,但本发明的实施方式并不受上述实施例的限制,其他的任何未背离本发明的精神实质与原理下所作的改变、修饰、替代、组合、简化,均应为等效的置换方式,都包含在本发明的保护范围之内。
Claims (10)
1.一种吡啶甲酸类生物碱,其特征在于,所述吡啶甲酸类生物碱具有式(I)、(II)结构:
2.权利要求1所述吡啶甲酸类生物碱的制备方法,其特征在于,所述吡啶甲酸类生物碱由南海海百合来源真菌发酵产物提取、分离、纯化得到。
3.根据权利要求2所述制备方法,其特征在于,所述南海海百合来源真菌命名为Penicillium brocae SYSU-CJ-17,保藏在广东省微生物菌种保藏中心,保藏日期是2023年05月26日,保藏号是GDMCC No:63444。
4.根据权利要求3所述制备方法,其特征在于,所述制备方法具体包括以下步骤:
S1、将南海海百合来源真菌活化、扩大培养,得到发酵物;
S2、将步骤S1所得发酵物用乙酸乙酯浸泡提取,提取液浓缩得到粗浸膏;
S3、将步骤S2所得粗浸膏进行硅胶柱层析,以石油醚-乙酸乙酯作为洗脱液进行梯度洗脱,收集石油醚-乙酸乙酯体积比为1:1的流份;
S4、将步骤S3所得石油醚-乙酸乙酯体积比为1:1的流份高效液相进行分离纯化,得到吡啶甲酸类生物碱。
5.根据权利要求4所述制备方法,其特征在于,步骤S3中,所述梯度洗脱为以石油醚-乙酸乙酯作为洗脱液,按照石油醚-乙酸乙酯体积比5:1、4:1、3:1、2:1、1:1、2:3、1:4、0:1依次进行梯度洗脱。
6.根据权利要求4所述制备方法,其特征在于,制备吡啶甲酸类生物碱化合物(I)时,步骤S4中的高效液相条件为流动相为50-70%甲醇/水,流速为2-3mL/min,检测波长为230nm或275nm,保留时间为10-20min。
7.根据权利要求4所述制备方法,其特征在于,制备吡啶甲酸类生物碱化合物(II)时,步骤S4中的高效液相条件为流动相为50-70%甲醇/水,流速为2-3mL/min,检测波长为230nm或275nm,保留时间为5-15min。
8.权利要求1所述吡啶甲酸类生物碱或其药学上可接受的盐在制备抗胃癌药物中的应用。
9.根据权利要求8所述应用,其特征在于,所述胃癌为MGC803、MKN45、HGC27、AGS、SNU1或KATO3胃癌细胞系造成的。
10.一种抗胃癌的药物组合物,其特征在于,所述组合物包括权利要求1所述吡啶甲酸类生物碱或其药学上可接受的盐和临床一线化疗药物。
Priority Applications (1)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
CN202311077059.6A CN117126105A (zh) | 2023-08-24 | 2023-08-24 | 一种吡啶甲酸类生物碱及其制备方法与应用 |
Applications Claiming Priority (1)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
CN202311077059.6A CN117126105A (zh) | 2023-08-24 | 2023-08-24 | 一种吡啶甲酸类生物碱及其制备方法与应用 |
Publications (1)
Publication Number | Publication Date |
---|---|
CN117126105A true CN117126105A (zh) | 2023-11-28 |
Family
ID=88859363
Family Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
CN202311077059.6A Pending CN117126105A (zh) | 2023-08-24 | 2023-08-24 | 一种吡啶甲酸类生物碱及其制备方法与应用 |
Country Status (1)
Country | Link |
---|---|
CN (1) | CN117126105A (zh) |
-
2023
- 2023-08-24 CN CN202311077059.6A patent/CN117126105A/zh active Pending
Similar Documents
Publication | Publication Date | Title |
---|---|---|
CN107353274B (zh) | 源于草酸青霉的黑麦酮酸i及制备抗人食管癌药物的应用 | |
CN107298672B (zh) | 源于草酸青霉的黑麦酮酸i在制备抗人结肠癌药物的应用 | |
CN107475146B (zh) | 一种链霉菌及其代谢产物粉蝶霉素类化合物在抗肾癌中的应用 | |
CN112592350A (zh) | 聚酮类化合物lithocarpin E-G及其制备方法和应用 | |
CN115806881A (zh) | 一种青霉属真菌及其在制备抗菌药物中的应用 | |
CN113801032B (zh) | 一种长链脂肪酸甘油醇类化合物Rubracin B、制备方法及其应用 | |
WO2013091361A1 (zh) | 环七肽及其在制备抗肿瘤药物中的应用 | |
CN105017203B (zh) | 一种来源于海洋真菌的Azaphilones类衍生化合物及其制备方法和应用 | |
CN112300243B (zh) | 一种环肽化合物及其制备方法和应用 | |
CN107298669B (zh) | 源于草酸青霉的黑麦酮酸i及抗人口腔表皮样癌药物应用 | |
CN117126105A (zh) | 一种吡啶甲酸类生物碱及其制备方法与应用 | |
CN113603594B (zh) | 倍半萜类化合物及其制备方法和在制备抗肿瘤药物中的应用 | |
CN111808112B (zh) | 一种Pratensilin D化合物及其制备和应用 | |
CN114716312A (zh) | 半日花烷型二萜化合物及其制备方法与应用 | |
CN109678917A (zh) | 艾美克勒霉素及其制备方法和应用 | |
CN114213428A (zh) | 一种吲哚生物碱化合物及其制备方法和应用 | |
CN109467579B (zh) | 一种具有免疫抑制活性的pks i型聚酮类化合物及其制备方法和应用 | |
CN109134417B (zh) | 源于草酸青霉的黑麦酮酸i及抗人子宫颈癌药物的应用 | |
CN116199695B (zh) | 一类桔霉素衍生物及其制备方法和应用 | |
CN108660169A (zh) | 一种发酵制备棘孢菌素类抗生素的方法 | |
CN111440200A (zh) | 一种混源萜生物碱及其抗寨卡病毒用途 | |
CN116041305B (zh) | 青霉菌(Penicillium mali)的发酵化合物及其制备方法和抗肿瘤应用 | |
CN110923278A (zh) | 源于草酸青霉的iso-Penicillixanthone A及在肺癌方面的应用 | |
CN109456196A (zh) | 一种海洋真菌来源的醌类化合物及其制备方法与应用 | |
CN115043790B (zh) | 一种对称杂环化合物、制备方法及其应用 |
Legal Events
Date | Code | Title | Description |
---|---|---|---|
PB01 | Publication | ||
PB01 | Publication | ||
SE01 | Entry into force of request for substantive examination | ||
SE01 | Entry into force of request for substantive examination |