CN111423448A - Stemona alkaloid skeleton-based small-molecule probe and preparation method and application thereof - Google Patents
Stemona alkaloid skeleton-based small-molecule probe and preparation method and application thereof Download PDFInfo
- Publication number
- CN111423448A CN111423448A CN202010264356.1A CN202010264356A CN111423448A CN 111423448 A CN111423448 A CN 111423448A CN 202010264356 A CN202010264356 A CN 202010264356A CN 111423448 A CN111423448 A CN 111423448A
- Authority
- CN
- China
- Prior art keywords
- furo
- azepine
- hexahydro
- methylene
- preparation
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Granted
Links
- 239000000523 sample Substances 0.000 title claims abstract description 26
- 241001671204 Stemona Species 0.000 title claims abstract description 22
- 150000003384 small molecules Chemical class 0.000 title claims abstract description 22
- 150000003797 alkaloid derivatives Chemical group 0.000 title claims abstract description 21
- 238000002360 preparation method Methods 0.000 title claims abstract description 20
- 238000006243 chemical reaction Methods 0.000 claims abstract description 20
- 150000001875 compounds Chemical class 0.000 claims abstract description 13
- 239000003068 molecular probe Substances 0.000 claims abstract description 13
- 206010009944 Colon cancer Diseases 0.000 claims abstract description 5
- 208000029742 colonic neoplasm Diseases 0.000 claims abstract description 5
- YMWUJEATGCHHMB-UHFFFAOYSA-N Dichloromethane Chemical compound ClCCl YMWUJEATGCHHMB-UHFFFAOYSA-N 0.000 claims description 72
- KAESVJOAVNADME-UHFFFAOYSA-N Pyrrole Chemical compound C=1C=CNC=1 KAESVJOAVNADME-UHFFFAOYSA-N 0.000 claims description 24
- ZMXDDKWLCZADIW-UHFFFAOYSA-N N,N-Dimethylformamide Chemical compound CN(C)C=O ZMXDDKWLCZADIW-UHFFFAOYSA-N 0.000 claims description 23
- 239000000243 solution Substances 0.000 claims description 19
- XKRFYHLGVUSROY-UHFFFAOYSA-N Argon Chemical compound [Ar] XKRFYHLGVUSROY-UHFFFAOYSA-N 0.000 claims description 18
- CTSLXHKWHWQRSH-UHFFFAOYSA-N oxalyl chloride Chemical compound ClC(=O)C(Cl)=O CTSLXHKWHWQRSH-UHFFFAOYSA-N 0.000 claims description 16
- QTBSBXVTEAMEQO-UHFFFAOYSA-N Acetic acid Chemical compound CC(O)=O QTBSBXVTEAMEQO-UHFFFAOYSA-N 0.000 claims description 12
- BKOOMYPCSUNDGP-UHFFFAOYSA-N 2-methylbut-2-ene Chemical compound CC=C(C)C BKOOMYPCSUNDGP-UHFFFAOYSA-N 0.000 claims description 10
- 238000004440 column chromatography Methods 0.000 claims description 10
- 238000001035 drying Methods 0.000 claims description 10
- OKKJLVBELUTLKV-UHFFFAOYSA-N Methanol Chemical compound OC OKKJLVBELUTLKV-UHFFFAOYSA-N 0.000 claims description 9
- 229910052786 argon Inorganic materials 0.000 claims description 9
- -1 prop-2-yne-1-amido Chemical group 0.000 claims description 9
- 238000003756 stirring Methods 0.000 claims description 8
- 239000007832 Na2SO4 Substances 0.000 claims description 7
- JKANAVGODYYCQF-UHFFFAOYSA-N prop-2-yn-1-amine Chemical compound NCC#C JKANAVGODYYCQF-UHFFFAOYSA-N 0.000 claims description 7
- 229910052938 sodium sulfate Inorganic materials 0.000 claims description 7
- VMHLLURERBWHNL-UHFFFAOYSA-M Sodium acetate Chemical compound [Na+].CC([O-])=O VMHLLURERBWHNL-UHFFFAOYSA-M 0.000 claims description 6
- GSEJCLTVZPLZKY-UHFFFAOYSA-N Triethanolamine Chemical compound OCCN(CCO)CCO GSEJCLTVZPLZKY-UHFFFAOYSA-N 0.000 claims description 6
- 229960000583 acetic acid Drugs 0.000 claims description 6
- 238000000605 extraction Methods 0.000 claims description 6
- 239000012362 glacial acetic acid Substances 0.000 claims description 6
- OOSZCNKVJAVHJI-UHFFFAOYSA-N 1-[(4-fluorophenyl)methyl]piperazine Chemical compound C1=CC(F)=CC=C1CN1CCNCC1 OOSZCNKVJAVHJI-UHFFFAOYSA-N 0.000 claims description 5
- 239000005708 Sodium hypochlorite Substances 0.000 claims description 5
- 239000013078 crystal Substances 0.000 claims description 5
- 229940074545 sodium dihydrogen phosphate dihydrate Drugs 0.000 claims description 5
- SUKJFIGYRHOWBL-UHFFFAOYSA-N sodium hypochlorite Chemical compound [Na+].Cl[O-] SUKJFIGYRHOWBL-UHFFFAOYSA-N 0.000 claims description 5
- RYHBNJHYFVUHQT-UHFFFAOYSA-N 1,4-Dioxane Chemical compound C1COCCO1 RYHBNJHYFVUHQT-UHFFFAOYSA-N 0.000 claims description 4
- ASOKPJOREAFHNY-UHFFFAOYSA-N 1-Hydroxybenzotriazole Chemical compound C1=CC=C2N(O)N=NC2=C1 ASOKPJOREAFHNY-UHFFFAOYSA-N 0.000 claims description 4
- FPQQSJJWHUJYPU-UHFFFAOYSA-N 3-(dimethylamino)propyliminomethylidene-ethylazanium;chloride Chemical compound Cl.CCN=C=NCCCN(C)C FPQQSJJWHUJYPU-UHFFFAOYSA-N 0.000 claims description 4
- 239000007864 aqueous solution Substances 0.000 claims description 4
- BDAGIHXWWSANSR-UHFFFAOYSA-N formic acid Substances OC=O BDAGIHXWWSANSR-UHFFFAOYSA-N 0.000 claims description 4
- 239000011541 reaction mixture Substances 0.000 claims description 4
- 239000001632 sodium acetate Substances 0.000 claims description 4
- 235000017281 sodium acetate Nutrition 0.000 claims description 4
- DXZDEAJXVCLRLE-UHFFFAOYSA-N azepin-2-one Chemical compound O=C1C=CC=CC=N1 DXZDEAJXVCLRLE-UHFFFAOYSA-N 0.000 claims description 3
- 239000012046 mixed solvent Substances 0.000 claims description 3
- 239000011734 sodium Substances 0.000 claims description 3
- IKAQCQLRDJKBSJ-GHMZBOCLSA-N (2R,6R)-3-methylidene-5-oxa-10-azatricyclo[8.3.0.02,6]trideca-1(13),11-dien-4-one Chemical compound C=C1C(O[C@H]2[C@H]1C=1N(CCC2)C=CC1)=O IKAQCQLRDJKBSJ-GHMZBOCLSA-N 0.000 claims description 2
- ZHNUHDYFZUAESO-UHFFFAOYSA-N formamide Substances NC=O ZHNUHDYFZUAESO-UHFFFAOYSA-N 0.000 claims description 2
- 230000035755 proliferation Effects 0.000 claims description 2
- 230000035484 reaction time Effects 0.000 claims 2
- 230000015572 biosynthetic process Effects 0.000 abstract description 5
- 238000003786 synthesis reaction Methods 0.000 abstract description 5
- 230000000694 effects Effects 0.000 abstract description 4
- 238000013461 design Methods 0.000 abstract description 3
- 150000002894 organic compounds Chemical class 0.000 abstract description 2
- 238000005457 optimization Methods 0.000 abstract 1
- XEKOWRVHYACXOJ-UHFFFAOYSA-N Ethyl acetate Chemical compound CCOC(C)=O XEKOWRVHYACXOJ-UHFFFAOYSA-N 0.000 description 12
- HEDRZPFGACZZDS-MICDWDOJSA-N Trichloro(2H)methane Chemical compound [2H]C(Cl)(Cl)Cl HEDRZPFGACZZDS-MICDWDOJSA-N 0.000 description 12
- 239000003814 drug Substances 0.000 description 10
- RTZKZFJDLAIYFH-UHFFFAOYSA-N Diethyl ether Chemical compound CCOCC RTZKZFJDLAIYFH-UHFFFAOYSA-N 0.000 description 8
- 102000004169 proteins and genes Human genes 0.000 description 8
- 108090000623 proteins and genes Proteins 0.000 description 8
- PEDCQBHIVMGVHV-UHFFFAOYSA-N Glycerine Chemical compound OCC(O)CO PEDCQBHIVMGVHV-UHFFFAOYSA-N 0.000 description 6
- 229940125782 compound 2 Drugs 0.000 description 6
- 229940079593 drug Drugs 0.000 description 6
- 229930014626 natural product Natural products 0.000 description 5
- 238000011160 research Methods 0.000 description 5
- 238000005160 1H NMR spectroscopy Methods 0.000 description 4
- 239000006166 lysate Substances 0.000 description 4
- 239000003208 petroleum Substances 0.000 description 4
- LNXBVCPZPORMEI-VXGBXAGGSA-N (2R,6R)-3-methylidene-4-oxo-5-oxa-10-azatricyclo[8.3.0.02,6]trideca-1(13),11-diene-11-carbaldehyde Chemical compound C=C1C(O[C@H]2[C@H]1C=1N(CCC2)C(=CC=1)C=O)=O LNXBVCPZPORMEI-VXGBXAGGSA-N 0.000 description 3
- IAZDPXIOMUYVGZ-UHFFFAOYSA-N Dimethylsulphoxide Chemical compound CS(C)=O IAZDPXIOMUYVGZ-UHFFFAOYSA-N 0.000 description 3
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 description 3
- 229940126214 compound 3 Drugs 0.000 description 3
- 229940125898 compound 5 Drugs 0.000 description 3
- 239000003085 diluting agent Substances 0.000 description 3
- 239000000203 mixture Substances 0.000 description 3
- 239000008188 pellet Substances 0.000 description 3
- 238000002264 polyacrylamide gel electrophoresis Methods 0.000 description 3
- YNJBWRMUSHSURL-UHFFFAOYSA-N trichloroacetic acid Chemical compound OC(=O)C(Cl)(Cl)Cl YNJBWRMUSHSURL-UHFFFAOYSA-N 0.000 description 3
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 3
- WVTKBKWTSCPRNU-KYJUHHDHSA-N (+)-Tetrandrine Chemical compound C([C@H]1C=2C=C(C(=CC=2CCN1C)OC)O1)C(C=C2)=CC=C2OC(=C2)C(OC)=CC=C2C[C@@H]2N(C)CCC3=CC(OC)=C(OC)C1=C23 WVTKBKWTSCPRNU-KYJUHHDHSA-N 0.000 description 2
- 238000001644 13C nuclear magnetic resonance spectroscopy Methods 0.000 description 2
- JKMHFZQWWAIEOD-UHFFFAOYSA-N 2-[4-(2-hydroxyethyl)piperazin-1-yl]ethanesulfonic acid Chemical compound OCC[NH+]1CCN(CCS([O-])(=O)=O)CC1 JKMHFZQWWAIEOD-UHFFFAOYSA-N 0.000 description 2
- YEDUAINPPJYDJZ-UHFFFAOYSA-N 2-hydroxybenzothiazole Chemical compound C1=CC=C2SC(O)=NC2=C1 YEDUAINPPJYDJZ-UHFFFAOYSA-N 0.000 description 2
- CURLTUGMZLYLDI-UHFFFAOYSA-N Carbon dioxide Chemical compound O=C=O CURLTUGMZLYLDI-UHFFFAOYSA-N 0.000 description 2
- KCXVZYZYPLLWCC-UHFFFAOYSA-N EDTA Chemical compound OC(=O)CN(CC(O)=O)CCN(CC(O)=O)CC(O)=O KCXVZYZYPLLWCC-UHFFFAOYSA-N 0.000 description 2
- 239000007995 HEPES buffer Substances 0.000 description 2
- UFHFLCQGNIYNRP-UHFFFAOYSA-N Hydrogen Chemical compound [H][H] UFHFLCQGNIYNRP-UHFFFAOYSA-N 0.000 description 2
- 238000005481 NMR spectroscopy Methods 0.000 description 2
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 description 2
- 239000012300 argon atmosphere Substances 0.000 description 2
- BLUAFEHZUWYNDE-NNWCWBAJSA-N artemisinin Chemical compound C([C@](OO1)(C)O2)C[C@H]3[C@H](C)CC[C@@H]4[C@@]31[C@@H]2OC(=O)[C@@H]4C BLUAFEHZUWYNDE-NNWCWBAJSA-N 0.000 description 2
- 229930101531 artemisinin Natural products 0.000 description 2
- 229960004191 artemisinin Drugs 0.000 description 2
- 239000000872 buffer Substances 0.000 description 2
- 229910002092 carbon dioxide Inorganic materials 0.000 description 2
- 238000005119 centrifugation Methods 0.000 description 2
- 229940125904 compound 1 Drugs 0.000 description 2
- 238000001514 detection method Methods 0.000 description 2
- 239000000975 dye Substances 0.000 description 2
- 239000007850 fluorescent dye Substances 0.000 description 2
- 239000001963 growth medium Substances 0.000 description 2
- 239000001257 hydrogen Substances 0.000 description 2
- 229910052739 hydrogen Inorganic materials 0.000 description 2
- NPZTUJOABDZTLV-UHFFFAOYSA-N hydroxybenzotriazole Substances O=C1C=CC=C2NNN=C12 NPZTUJOABDZTLV-UHFFFAOYSA-N 0.000 description 2
- 230000002401 inhibitory effect Effects 0.000 description 2
- 238000000034 method Methods 0.000 description 2
- 239000000137 peptide hydrolase inhibitor Substances 0.000 description 2
- 239000000256 polyoxyethylene sorbitan monolaurate Substances 0.000 description 2
- 238000001228 spectrum Methods 0.000 description 2
- 239000012192 staining solution Substances 0.000 description 2
- 239000000725 suspension Substances 0.000 description 2
- 238000001308 synthesis method Methods 0.000 description 2
- 229940126680 traditional chinese medicines Drugs 0.000 description 2
- 238000005406 washing Methods 0.000 description 2
- LBAGJGNPTDWYFK-UHFFFAOYSA-N 3h-pyrrolo[1,2-a]azepine-8-carboxamide Chemical compound NC(=O)C1=CC=CN2CC=CC2=C1 LBAGJGNPTDWYFK-UHFFFAOYSA-N 0.000 description 1
- 238000009010 Bradford assay Methods 0.000 description 1
- HEDRZPFGACZZDS-UHFFFAOYSA-N CHCl3 Substances ClC(Cl)Cl HEDRZPFGACZZDS-UHFFFAOYSA-N 0.000 description 1
- OKTJSMMVPCPJKN-UHFFFAOYSA-N Carbon Chemical compound [C] OKTJSMMVPCPJKN-UHFFFAOYSA-N 0.000 description 1
- 239000006144 Dulbecco’s modified Eagle's medium Substances 0.000 description 1
- 241001115402 Ebolavirus Species 0.000 description 1
- LSDPWZHWYPCBBB-UHFFFAOYSA-N Methanethiol Chemical compound SC LSDPWZHWYPCBBB-UHFFFAOYSA-N 0.000 description 1
- IPQKDIRUZHOIOM-UHFFFAOYSA-N Oroxin A Natural products OC1C(O)C(O)C(CO)OC1OC(C(=C1O)O)=CC2=C1C(=O)C=C(C=1C=CC=CC=1)O2 IPQKDIRUZHOIOM-UHFFFAOYSA-N 0.000 description 1
- KDLHZDBZIXYQEI-UHFFFAOYSA-N Palladium Chemical compound [Pd] KDLHZDBZIXYQEI-UHFFFAOYSA-N 0.000 description 1
- 108010019160 Pancreatin Proteins 0.000 description 1
- 229920001213 Polysorbate 20 Polymers 0.000 description 1
- 229940124158 Protease/peptidase inhibitor Drugs 0.000 description 1
- DFBIRQPKNDILPW-CIVMWXNOSA-N Triptolide Chemical compound O=C1OCC([C@@H]2C3)=C1CC[C@]2(C)[C@]12O[C@H]1[C@@H]1O[C@]1(C(C)C)[C@@H](O)[C@]21[C@H]3O1 DFBIRQPKNDILPW-CIVMWXNOSA-N 0.000 description 1
- 239000007983 Tris buffer Substances 0.000 description 1
- 102000004142 Trypsin Human genes 0.000 description 1
- 108090000631 Trypsin Proteins 0.000 description 1
- 239000002250 absorbent Substances 0.000 description 1
- 230000002745 absorbent Effects 0.000 description 1
- 238000009825 accumulation Methods 0.000 description 1
- 239000003242 anti bacterial agent Substances 0.000 description 1
- 230000000078 anti-malarial effect Effects 0.000 description 1
- 229940088710 antibiotic agent Drugs 0.000 description 1
- 239000003430 antimalarial agent Substances 0.000 description 1
- 239000012298 atmosphere Substances 0.000 description 1
- IKIIZLYTISPENI-ZFORQUDYSA-N baicalin Chemical compound O1[C@H](C(O)=O)[C@@H](O)[C@H](O)[C@@H](O)[C@@H]1OC(C(=C1O)O)=CC2=C1C(=O)C=C(C=1C=CC=CC=1)O2 IKIIZLYTISPENI-ZFORQUDYSA-N 0.000 description 1
- AQHDANHUMGXSJZ-UHFFFAOYSA-N baicalin Natural products OC1C(O)C(C(O)CO)OC1OC(C(=C1O)O)=CC2=C1C(=O)C=C(C=1C=CC=CC=1)O2 AQHDANHUMGXSJZ-UHFFFAOYSA-N 0.000 description 1
- 229960003321 baicalin Drugs 0.000 description 1
- 230000004071 biological effect Effects 0.000 description 1
- 229910052799 carbon Inorganic materials 0.000 description 1
- 239000001569 carbon dioxide Substances 0.000 description 1
- 239000006143 cell culture medium Substances 0.000 description 1
- 230000004663 cell proliferation Effects 0.000 description 1
- 238000012650 click reaction Methods 0.000 description 1
- 238000006482 condensation reaction Methods 0.000 description 1
- 238000012790 confirmation Methods 0.000 description 1
- 238000012258 culturing Methods 0.000 description 1
- 238000002349 difference gel electrophoresis Methods 0.000 description 1
- 238000007865 diluting Methods 0.000 description 1
- 229940042399 direct acting antivirals protease inhibitors Drugs 0.000 description 1
- 238000001962 electrophoresis Methods 0.000 description 1
- 238000002474 experimental method Methods 0.000 description 1
- 239000012145 high-salt buffer Substances 0.000 description 1
- 238000011534 incubation Methods 0.000 description 1
- 229940042040 innovative drug Drugs 0.000 description 1
- 150000002611 lead compounds Chemical class 0.000 description 1
- 230000003287 optical effect Effects 0.000 description 1
- 230000001590 oxidative effect Effects 0.000 description 1
- 229940055695 pancreatin Drugs 0.000 description 1
- 230000000144 pharmacologic effect Effects 0.000 description 1
- 235000010486 polyoxyethylene sorbitan monolaurate Nutrition 0.000 description 1
- 239000002244 precipitate Substances 0.000 description 1
- 238000002731 protein assay Methods 0.000 description 1
- 238000011002 quantification Methods 0.000 description 1
- WVTKBKWTSCPRNU-UHFFFAOYSA-N rac-Tetrandrin Natural products O1C(C(=CC=2CCN3C)OC)=CC=2C3CC(C=C2)=CC=C2OC(=C2)C(OC)=CC=C2CC2N(C)CCC3=CC(OC)=C(OC)C1=C23 WVTKBKWTSCPRNU-UHFFFAOYSA-N 0.000 description 1
- 239000002994 raw material Substances 0.000 description 1
- 239000011780 sodium chloride Substances 0.000 description 1
- 238000002415 sodium dodecyl sulfate polyacrylamide gel electrophoresis Methods 0.000 description 1
- 238000005556 structure-activity relationship Methods 0.000 description 1
- 239000006228 supernatant Substances 0.000 description 1
- YKUJZZHGTWVWHA-UHFFFAOYSA-N triptolide Natural products COC12CC3OC3(C(C)C)C(O)C14OC4CC5C6=C(CCC25C)C(=O)OC6 YKUJZZHGTWVWHA-UHFFFAOYSA-N 0.000 description 1
- LENZDBCJOHFCAS-UHFFFAOYSA-N tris Chemical compound OCC(N)(CO)CO LENZDBCJOHFCAS-UHFFFAOYSA-N 0.000 description 1
- 239000012588 trypsin Substances 0.000 description 1
- 238000012795 verification Methods 0.000 description 1
- 239000012130 whole-cell lysate Substances 0.000 description 1
Images
Classifications
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07D—HETEROCYCLIC COMPOUNDS
- C07D491/00—Heterocyclic compounds containing in the condensed ring system both one or more rings having oxygen atoms as the only ring hetero atoms and one or more rings having nitrogen atoms as the only ring hetero atoms, not provided for by groups C07D451/00 - C07D459/00, C07D463/00, C07D477/00 or C07D489/00
- C07D491/12—Heterocyclic compounds containing in the condensed ring system both one or more rings having oxygen atoms as the only ring hetero atoms and one or more rings having nitrogen atoms as the only ring hetero atoms, not provided for by groups C07D451/00 - C07D459/00, C07D463/00, C07D477/00 or C07D489/00 in which the condensed system contains three hetero rings
- C07D491/14—Ortho-condensed systems
- C07D491/147—Ortho-condensed systems the condensed system containing one ring with oxygen as ring hetero atom and two rings with nitrogen as ring hetero atom
-
- 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
- C09—DYES; PAINTS; POLISHES; NATURAL RESINS; ADHESIVES; COMPOSITIONS NOT OTHERWISE PROVIDED FOR; APPLICATIONS OF MATERIALS NOT OTHERWISE PROVIDED FOR
- C09K—MATERIALS FOR MISCELLANEOUS APPLICATIONS, NOT PROVIDED FOR ELSEWHERE
- C09K11/00—Luminescent, e.g. electroluminescent, chemiluminescent materials
- C09K11/06—Luminescent, e.g. electroluminescent, chemiluminescent materials containing organic luminescent materials
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N1/00—Sampling; Preparing specimens for investigation
- G01N1/28—Preparing specimens for investigation including physical details of (bio-)chemical methods covered elsewhere, e.g. G01N33/50, C12Q
- G01N1/30—Staining; Impregnating ; Fixation; Dehydration; Multistep processes for preparing samples of tissue, cell or nucleic acid material and the like for analysis
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N1/00—Sampling; Preparing specimens for investigation
- G01N1/28—Preparing specimens for investigation including physical details of (bio-)chemical methods covered elsewhere, e.g. G01N33/50, C12Q
- G01N1/40—Concentrating samples
- G01N1/4055—Concentrating samples by solubility techniques
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
- G01N21/62—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light
- G01N21/63—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light optically excited
- G01N21/64—Fluorescence; Phosphorescence
- G01N21/6486—Measuring fluorescence of biological material, e.g. DNA, RNA, cells
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N27/00—Investigating or analysing materials by the use of electric, electrochemical, or magnetic means
- G01N27/26—Investigating or analysing materials by the use of electric, electrochemical, or magnetic means by investigating electrochemical variables; by using electrolysis or electrophoresis
- G01N27/416—Systems
- G01N27/447—Systems using electrophoresis
- G01N27/44704—Details; Accessories
- G01N27/44747—Composition of gel or of carrier mixture
-
- C—CHEMISTRY; METALLURGY
- C09—DYES; PAINTS; POLISHES; NATURAL RESINS; ADHESIVES; COMPOSITIONS NOT OTHERWISE PROVIDED FOR; APPLICATIONS OF MATERIALS NOT OTHERWISE PROVIDED FOR
- C09K—MATERIALS FOR MISCELLANEOUS APPLICATIONS, NOT PROVIDED FOR ELSEWHERE
- C09K2211/00—Chemical nature of organic luminescent or tenebrescent compounds
- C09K2211/10—Non-macromolecular compounds
- C09K2211/1018—Heterocyclic compounds
- C09K2211/1025—Heterocyclic compounds characterised by ligands
- C09K2211/1029—Heterocyclic compounds characterised by ligands containing one nitrogen atom as the heteroatom
- C09K2211/1033—Heterocyclic compounds characterised by ligands containing one nitrogen atom as the heteroatom with oxygen
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N1/00—Sampling; Preparing specimens for investigation
- G01N1/28—Preparing specimens for investigation including physical details of (bio-)chemical methods covered elsewhere, e.g. G01N33/50, C12Q
- G01N1/40—Concentrating samples
- G01N1/4055—Concentrating samples by solubility techniques
- G01N2001/4061—Solvent extraction
Landscapes
- Chemical & Material Sciences (AREA)
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Organic Chemistry (AREA)
- General Health & Medical Sciences (AREA)
- Pathology (AREA)
- Immunology (AREA)
- Molecular Biology (AREA)
- Physics & Mathematics (AREA)
- Analytical Chemistry (AREA)
- Biochemistry (AREA)
- General Physics & Mathematics (AREA)
- Engineering & Computer Science (AREA)
- Biomedical Technology (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
- Materials Engineering (AREA)
- General Chemical & Material Sciences (AREA)
- Medicinal Chemistry (AREA)
- Pharmacology & Pharmacy (AREA)
- Animal Behavior & Ethology (AREA)
- Public Health (AREA)
- Veterinary Medicine (AREA)
- Dispersion Chemistry (AREA)
- Electrochemistry (AREA)
- Nitrogen Condensed Heterocyclic Rings (AREA)
Abstract
The invention particularly relates to a small molecular probe based on a stemona alkaloid skeleton, a preparation method and application thereof, belonging to the technical field of organic compounds and preparation thereof. The invention completes the design and synthesis of a small molecular probe (positive probe) based on the compound SA-11 structure, the small molecular probe can be used together with a bioorthogonal reaction, the effect target of an active small molecule based on the stemona alkaloid skeleton is found, the foundation is laid for the subsequent optimization of the small molecular structure, and the small molecular probe can also inhibit colon cancer cells HCT-116.
Description
Technical Field
The invention particularly relates to a small molecular probe based on a stemona alkaloid skeleton, a preparation method and application thereof, belonging to the technical field of organic compounds and preparation thereof.
Background
The traditional Chinese medicine and the natural medicine are treasures of medicine discovery, and the ever anti-malarial artemisinin and the newly discovered tetrandrine capable of resisting the Ebola virus are both derived from the traditional Chinese medicine. Under the background of high research cost, long time and increasing difficulty of innovative drugs, the search of lead compounds from traditional Chinese medicines and natural medicines is again a hot spot in recent years. Natural products which can be separated from traditional Chinese medicines and natural medicines have remarkable biological activity, but the action targets of the natural products are unknown, and the small yield of the natural products also limits further pharmacological mechanism research. The research on the effective new research method has important significance.
Currently, various strategies are developed and applied to target search of active natural products, wherein the strategy based on active-based Protein Profiling (ABPP) is widely applied to target search of active small molecules. The application of multiple natural product star molecules has completed the confirmation of target points, such as adenitin, triptolide, HEP14, artemisinin, baicalin, etc. Whereas the ABPP strategy first step is to construct probes based on active small molecule structures. In the previous research, we found that small molecules based on stemona alkaloid skeleton have significant activity of inhibiting colon cancer, but the action mechanism is not clear. Therefore, on the basis of accumulation of the former-stage synthesis method and structure-activity relationship, design and synthesis of a positive probe based on the structure of the compound SA-11 are completed, and in order to exclude non-specific protein binding in the target fishing process, design and synthesis of a negative probe corresponding to the positive probe are completed at the same time, and activity verification is completed. In the pull-down experiment, the target proteins were separated by SDS-PAGE electrophoresis, and several distinct protein bands were found compared to the negative probe.
The structural formula of the compound SA-11 is as follows:
disclosure of Invention
The invention aims to provide a small molecular probe based on a stemona alkaloid skeleton, a synthesis method and application thereof.
In order to achieve the purpose, the invention adopts the following technical scheme:
a small molecule probe (positive probe) based on stemona alkaloid skeleton is (3aR,10bR) -1-methylene-2-oxygen-N- (prop-2-alkyne-1-amido) -1,3a,4,5,6,10 b-hexahydro-2H-furo [3,2-c ] pyrrole [1,2-a ] azepine-8-formamide, and the structural formula is as follows:
the preparation method of the molecular probe comprises the following steps:
(3aR,10bR) -1-methylene-1, 3a,4,5,6,10 b-hexahydro-2H-furo [3,2-c ] pyrrolo [1,2-a ] azepine-2-one is reacted with oxalyl chloride and dimethylformamide to form (3aR,10bR) -1-methylene-2-oxo-1, 3a,4,5,6,10 b-hexahydro-2H-furo [3,2-c ] pyrrolo [1,2-a ] azepine-8-carbaldehyde, (3aR,10bR) -1-methylene-2-oxo-1, 3a,4,5,6,10 b-hexahydro-2H-furo [3,2-c ] pyrrolo [1,2-a ] azepine-8-carbaldehyde Oxidizing with the following to generate (3aR,10bR) -1-methylene-2-oxo-1, 3a,4,5,6,10 b-hexahydro-2H-furo [3,2-c ] pyrrole [1,2-a ] azepine-8-formic acid, and carrying out condensation reaction on the (3aR,10bR) -1-methylene-2-oxo-1, 3a,4,5,6,10 b-hexahydro-2H-furo [3,2-c ] pyrrole [1,2-a ] azepine-8-formic acid and propargylamine to generate (3aR,10bR) -1-methylene-2-oxo-N- (prop-2-yne-1-amino) -1,3a,4,5,6,10 b-hexahydro-2H-furo [3,2-c ] pyrrolo [1,2-a ] azepine-8-carboxamide.
A preparation method of a stemona alkaloid skeleton-based small molecule probe comprises the following steps:
step 1, oxalyl chloride is added dropwise to dry dimethylformamide under argon protection, the reaction is continued under these conditions after white crystals have formed, anTo this mixture was added dichloromethane followed by (3aR,10bR) -1-methylene-1, 3a,4,5,6,10 b-hexahydro-2H-furo [3,2-c ]]Pyrrole [1,2-a ]]Adding sodium acetate aqueous solution after continuing the reaction of the dichloromethane solution of the azepine-2-ketone (compound 1), stirring, stopping the reaction, extracting with dichloromethane, adding anhydrous Na2SO4Drying, and separating by column chromatography to obtain (3aR,10bR) -1-methylene-2-oxo-1, 3a,4,5,6,10 b-hexahydro-2H-furo [3, 2-c)]Pyrrole [1,2-a ]]Azepine-8-carbaldehyde (compound 3);
step 3, under the protection of argon, propargylamine, triethanolamine, 1-hydroxybenzotriazole and 1- (3-dimethylaminopropyl) -3-ethylcarbodiimide hydrochloride are sequentially added into (3aR,10bR) -1-methylene-2-oxo-1, 3a,4,5,6,10 b-hexahydro-2H-furo [3,2-c ] at room temperature]Pyrrole [1,2-a ]]The azepine-8-formic acid is put into dichloromethane solution and reacts at room temperature overnight, and then dichloromethane is used for extraction, anhydrous Na is used2SO4Drying and separating by column chromatography to obtain (3aR,10bR) -1-methylene-2-oxygen-N- (prop-2-alkyne-1-amido) -1,3a,4,5,6,10 b-hexahydro-2H-furo [3, 2-c)]Pyrrole [1,2-a ]]Azepine-8-carboxamide (Compound 2).
The route realizes the efficient synthesis of the stemona alkaloid skeleton-based active small molecular probe.
Further, in the step 1, oxalyl chloride is dropwise added into dry dimethylformamide at a reaction temperature of 0-10 ℃, the time for continuing the reaction after white crystals are generated is 15min, the time for continuing the reaction after (3aR,10bR) -1-methylene-1, 3a,4,5,6,10 b-hexahydro-2H-furo [3,2-c ] pyrrole [1,2-a ] azepine-2-one is added for 0.5H, the stirring time is 0.5H, and the extraction times are 3 times.
Further, in the step 1, the molar ratio of dimethylformamide to oxalyl chloride is 2-5: 2-5, the volume of dichloromethane is 8-16 m L, the concentration of a dichloromethane solution of (3aR,10bR) -1-methylene-1, 3a,4,5,6,10 b-hexahydro-2H-furo [3,2-c ] pyrrole [1,2-a ] azepine-2-one is 0.2-0.8 mol/L, the volume is 2-8 m L, the concentration of a sodium acetate aqueous solution is 1-3 mol/L, and the volume is 2-10 m L.
Further, in the step 2, the molar ratio of the sodium dihydrogen phosphate dihydrate to the sodium hypochlorite to the 2-methyl-2-butene is 4-10: 4-10, and the concentration of the 1, 4-dioxane solution of the (3aR,10bR) -1-methylene-2-oxo-1, 3a,4,5,6,10 b-hexahydro-2H-furo [3,2-c ] pyrrolo [1,2-a ] azepine-8-formaldehyde is 0.01-1 mol/L, and the volume is 1-5 m L.
Further, the stirring temperature in the step 2 is 28 ℃, the amount of glacial acetic acid is 2-5 m L, and the volume ratio of dichloromethane to methanol is 10: 1-20: 1.
In the step 3, the molar ratio of propargylamine, triethanolamine, 1-hydroxybenzotriazole and 1- (3-dimethylaminopropyl) -3-ethylcarbodiimide hydrochloride is 1.5-5: 10-15: 1.5-5, and the dichloromethane solution of (3aR,10bR) -1-methylene-2-oxo-1, 3a,4,5,6,10 b-hexahydro-2H-furo [3,2-c ] pyrrolo [1,2-a ] azepine-8-carboxylic acid has the concentration of 0.02-1 mol/L, the volume of 0.5-2 m L and the extraction times of 3 times.
An application of a small molecular probe based on radix Stemonae alkaloid skeleton can be used together with bioorthogonal reaction to perform effect target fishing of radix Stemonae alkaloid skeleton compound; can inhibit HCT-116 of colon cancer cells.
Compared with the existing fluorescent probe for detecting the micromolecule mercaptan, the fluorescent probe has the following advantages:
1. the invention provides a simple synthesis route of a small molecular probe based on a stemona alkaloid skeleton, and the probe is easy to obtain raw materials and low in cost;
2. the invention provides application of a small molecular probe, which can be used together with bioorthogonal reaction to perform action target fishing of stemona alkaloid skeleton compounds.
Drawings
FIG. 1 is a hydrogen spectrum of nuclear magnetic resonance of a small molecule probe;
FIG. 2 is a carbon spectrum of nuclear magnetic resonance of a small molecule probe;
FIG. 3 is data of inhibitory activity of small molecule probes on colon cancer cells HCT-116;
FIG. 4 is polyacrylamide gel electrophoresis of a target protein of a small molecule probe.
Detailed Description
Example 1
A preparation method of a stemona alkaloid skeleton-based small molecule probe comprises the following steps:
1) preparation of Compound 3
Oxalyl chloride (40 μ L) was added dropwise to dry DMF (72 μ L) under argon protection at 10 ℃ under conditions to give white crystals after which the reaction was continued for 15min, and 8m L DCM was added to the mixture, followed by 2m L solution of compound 1(95mg, 0.47mmo L) in DCM, and after 0.5h of further reaction an aqueous AcONa solution (225mg AcONa in 2.5m L water) was added, stirred for 0.5h, the reaction was stopped, extracted three times with DCM and anhydrous Na was added2SO4Compound 3(64.2mg) was isolated by column chromatography (petroleum ether: ethyl acetate ═ 2:1) with a yield of 59%.
As shown in FIG. 1:
1H NMR(600MHz,CDCl3)9.51(s,1H),6.90(d,J＝3.8Hz,1H),6.60(d,J＝3.3Hz,1H),6.25(d,J＝3.8Hz,1H),6.02(d,J＝2.4Hz,1H),5.84(dd,J＝14.8,5.2Hz,1H),4.17–4.10(m,1H),4.01–3.94(m,1H),3.81–3.73(m,1H),2.59(dd,J＝12.9,3.1Hz,1H),2.24–2.18(m,1H),1.92(qd,J＝13.0,3.3Hz,1H),1.68(d,J＝12.2Hz,1H).
as shown in fig. 2:
13C NMR(151MHz,CDCl3)179.89(s),169.12(s),138.74(s),134.35(s),132.24(s),124.85(s),123.96(s),107.01(s),80.64(s),45.07(s),44.78(s),34.26(s),25.72(s).
2) preparation of Compound 4
Sodium dihydrogen phosphate dihydrate (131mg, 0.84mmo L), sodium hypochlorite (76mg, 0.84mmo L) and 2-methyl-2-butene (90. mu.l, 0.84mmo L) were added to a solution of compound 3(32mg, 0.14mmo L) in 1, 4-dioxane (2m L) in this order at room temperature under argon atmosphere, the reaction was stirred overnight at 28 ℃, 2m L glacial acetic acid was added to the reaction mixture, the mixture was extracted with a mixed solvent of dichloromethane, methanol, 10:1 and anhydrous Na2SO4After drying, column chromatography (petroleum ether: ethyl acetate ═ 2:1) gave compound 4(20mg) in 58% yield.
1H NMR(600MHz,CDCl3)7.10(d,J＝3.9Hz,1H),6.60(d,J＝3.4Hz,1H),6.19(d,J＝3.5Hz,1H),6.04(d,J＝3.0Hz,1H),5.77(dd,J＝14.8,5.0Hz,1H),4.14(d,J＝7.2Hz,1H),4.02–3.95(m,1H),3.76(t,J＝13.1Hz,1H),2.61–2.56(m,1H),2.22(d,J＝11.8Hz,1H),1.92(d,J＝14.9Hz,1H),1.69(d,J＝13.6Hz,1H).
3) Preparation of Compound 2
Propargylamine (6 μ L, 0.09mmo L), TEA (43 μ L, 0.8mmo L), HOBT (12mg, 0.09mmo L) and EDC (17.2mg, 0.09mmo L) were added to Compound 4(15mg, 0.06mmo L) in this order under argon at room temperature) In dichloromethane (1m L), overnight at room temperature, extracted three times with dichloromethane and dried over Na2SO4Drying and column chromatography (petroleum ether: ethyl acetate ═ 2:1) gave compound 2(14.5mg) in 85% yield.
1H NMR(600MHz,CDCl3)6.96(d,J＝4.1Hz,1H),6.58(d,J＝3.6Hz,1H),6.14(d,J＝4.0Hz,1H),6.02(d,J＝3.2Hz,1H),5.77(dd,J＝14.9,5.7Hz,1H),5.44–5.34(m,2H),4.15–4.08(m,1H),4.00–3.93(m,1H),3.76(d,J＝7.1Hz,2H),2.60(s,3H),2.28–2.18(m,1H),1.91(qd,J＝13.3,4.2Hz,1H),1.66(dt,J＝23.8,11.1Hz,2H).
13C NMR(126MHz,CDCl3)178.9,171.6,133.4,128.7,113.1,106.9,103.0,47.3,45.2,36.1,35.36,34.7,19.9,15.0,13.1,10.6.
Example 2
1) Preparation of Compound 5
Palladium/carbon was added to a solution of compound 4(20mg, 0.08mmo L) in ethanol (2.5m L) under argon atmosphere, purged three times, hydrogen was introduced, the reaction was carried out at room temperature for 3 hours, filtered and concentrated to give compound 5(16mg) in 80% yield, which was used for the next reaction.
2) Preparation of Compound 6
Propargylamine (6 μ L, 0.09mmo L), TEA (43 μ L, 0.8mmo L), HOBT (12mg, 0.09mmo L) and EDC (17.2mg, 0.09mmo L) were added in succession to a solution of Compound 5(15mg, 0.06mmo L) in dichloromethane (1m L) under argon protection at room temperature, the reaction mixture was stirred overnight at room temperature, extracted three times with dichloromethane and dried over Na2SO4Drying and column chromatography (petroleum ether: ethyl acetate ═ 2:1) gave compound 6(13.7mg) in 80% yield.
1H NMR(600MHz,CDCl3)6.93(d,J＝4.0Hz,1H),5.99(d,J＝3.8Hz,1H),5.79(dd,J＝14.7,5.8Hz,1H),4.23(t,J＝6.6Hz,2H),4.14(d,J＝7.1Hz,1H),3.96–3.89(m,1H),3.75(d,J＝7.0Hz,1H),3.64(dd,J＝14.6,11.5Hz,1H),3.13–3.06(m,1H),3.05–2.98(m,1H),2.57(dd,J＝12.9,3.2Hz,1H),2.07(s,1H),1.97(s,1H),1.45(d,J＝6.8Hz,3H),1.30–1.26(m,1H).
Example 3
detecting cell proliferation by SRB detection method, determining cell density based on protein content, digesting with pancreatin, and adjusting cell density to 4-6 × 10 with fresh culture medium4Pieces/m L, seeded in 96-well plates at 100. mu. L per well, placed at 37.0 ℃ in 5.0% CO2Diluting the drug to 2 times detection concentration by using a culture medium to ensure that the DMSO content in the drug diluent is consistent, sucking 100 mu L drug diluent and adding the drug diluent into a 96-well plate, continuously culturing for 72h, discarding the cell culture medium, lightly adding 10% (Wt/Vol) trichloroacetic acid (TCA) precooled by 100 mu L, standing at 4 ℃ for at least more than 1h, removing the TCA fixing solution, washing for five times by using slow flowing water, removing residual water by using absorbent paper, lightly adding 100 mu L SRB staining solution, incubating for 25min at room temperature, removing the SRB staining solution, washing for five times by using 1% glacial acetic acid solution to remove unbound dye, drying at room temperature, adding 100 mu L mM Tris (pH10.0) solution, and detecting OD (optical density) by using a microplate reader after the dye is completely dissolved570nm。
Example 4
Polyacrylamide gel electrophoresis of the target protein (see fig. 4):
HCT-116 cells were grown in DMEM supplemented with 10% FBS and no antibiotics in a humidified environment with 10% carbon dioxide added to an atmosphere at 37 ℃. Cells were approximately 80-90% confluent and treated with 1X PBS and trypsin. Cell pellets were obtained by centrifugation at 1000rpm for 5 minutes at 4 ℃ and the pellets were washed 3 times with 1 XPBS. To prepare a whole cell lysate, cells are first suspended in hypotonic buffer (10mM HEPES, pH 7.5, 2mM MgCl)20.1 % Tween 20, 20% glycerol, Roche completely absentEDTA-containing protease inhibitors) were incubated at 4 ℃ for 10 minutes. The suspension was separated after centrifugation at 16000 rpm for 15 minutes at 4 ℃. The pellet was then resuspended in high salt buffer (50mM HEPES, pH 7.5, 420mM NaCl, 2mM MgCl)2Tween-20 0.1%, glycerol 20%, protease inhibitor complete without EDTA roche.) incubation at 4 ℃ for 30 min the suspension was centrifuged at 16000 rpm for 15min at 4 ℃ the supernatant was collected and then combined with the soluble fraction in hypotonic buffer to give a lysate of whole cells after quantification of the protein using the Bradford assay (Bio-Rad DCTM protein assay), the lysate was labeled with compound 2 and compound 6. briefly, 100 μ g lysate was at a concentration of 1 μ g/μ L in the presence or absence of excess, then CuAAC catalyzed lysate was added, compound 2 and compound 6 with Biotin-PEG4-N3Click reaction was performed for 1.5 hours, quenched and added 10mM EDTA, with CHCl3The precipitate was then solubilized to 10mg/m L in PBS containing 2% SDS and 10mM DTT and finally diluted to 1mg/m L1 XPBS the proteins were separated on a 12% SDS gel using PAGE and fluorescence was obtained by scanning the gel using an Ettan. TM. DIGE imager (GE Healthcare).
Claims (8)
1. The small molecule probe based on the stemona alkaloid skeleton is characterized in that the small molecule probe is (3aR,10bR) -1-methylene-2-oxy-N- (prop-2-yne-1-amido) -1,3a,4,5,6,10 b-hexahydro-2H-furo [3,2-c ] pyrrole [1,2-a ] azepine-8-formamide, and the structural formula of the small molecule probe is as follows:
2. a preparation method of a stemona alkaloid skeleton-based small molecule probe is characterized by comprising the following steps:
step 1, oxalyl chloride was added dropwise to dry dimethylformamide under argon protection, the reaction was continued under these conditions after white crystals were produced, and this was mixedDichloromethane is added to the compound, then (3aR,10bR) -1-methylene-1, 3a,4,5,6,10 b-hexahydro-2H-furo [3,2-c ] is added]Pyrrole [1,2-a ]]Continuing the reaction of the solution of azepine-2-ketone in dichloromethane, adding aqueous solution of sodium acetate, stirring, stopping the reaction, extracting with dichloromethane, and adding anhydrous Na2SO4Drying, and separating by column chromatography to obtain (3aR,10bR) -1-methylene-2-oxo-1, 3a,4,5,6,10 b-hexahydro-2H-furo [3, 2-c)]Pyrrole [1,2-a ]]Azepine-8-carbaldehyde;
step 2, adding sodium dihydrogen phosphate dihydrate, sodium hypochlorite and 2-methyl-2-butene into (3aR,10bR) -1-methylene-2-oxo-1, 3a,4,5,6,10 b-hexahydro-2H-furo [3,2-c ] in sequence at room temperature under the protection of argon]Pyrrole [1,2-a ]]Adding 1, 4-dioxane solution of azepine-8-formaldehyde, stirring overnight, adding glacial acetic acid into the reaction mixture, extracting with mixed solvent, and adding anhydrous Na2SO4Drying and separating by column chromatography to obtain (3aR,10bR) -1-methylene-2-oxygen-1, 3a,4,5,6,10 b-hexahydro-2H-furo [3, 2-c)]Pyrrole [1,2-a ]]Azepine-8-carboxylic acid;
step 3, under the protection of argon, propargylamine, triethanolamine, 1-hydroxybenzotriazole and 1- (3-dimethylaminopropyl) -3-ethylcarbodiimide hydrochloride are sequentially added into (3aR,10bR) -1-methylene-2-oxo-1, 3a,4,5,6,10 b-hexahydro-2H-furo [3,2-c ] at room temperature]Pyrrole [1,2-a ]]The azepine-8-formic acid is put into dichloromethane solution and reacts at room temperature overnight, and then dichloromethane is used for extraction, anhydrous Na is used2SO4Drying and separating by column chromatography to obtain (3aR,10bR) -1-methylene-2-oxygen-N- (prop-2-alkyne-1-amido) -1,3a,4,5,6,10 b-hexahydro-2H-furo [3, 2-c)]Pyrrole [1,2-a ]]Azepine-8-carboxamide.
3. The preparation method of the stemona alkaloid skeleton-based small molecule probe as claimed in claim 2, wherein the reaction temperature of dropwise adding oxalyl chloride into dry dimethylformamide in step 1 is 0-10 ℃, the reaction time after white crystals are generated is 15min, the reaction time after adding (3aR,10bR) -1-methylene-1, 3a,4,5,6,10 b-hexahydro-2H-furo [3,2-c ] pyrrole [1,2-a ] azepine-2-one is 0.5H, the stirring time is 0.5H, and the extraction times are 3 times.
4. The preparation method of the stemona alkaloid skeleton-based small molecule probe as claimed in claim 2, wherein the molar ratio of dimethylformamide to oxalyl chloride in step 1 is 2-5: 2-5, the volume of dichloromethane is 8-16 m L, the concentration of dichloromethane solution of (3aR,10bR) -1-methylene-1, 3a,4,5,6,10 b-hexahydro-2H-furo [3,2-c ] pyrrolo [1,2-a ] azepine-2-one is 0.2-0.8 mol/L, the volume is 2-8 m L, the concentration of sodium acetate aqueous solution is 1-3 mol/L, and the volume is 2-10 m L.
5. The preparation method of the stemona alkaloid skeleton-based small molecule probe as claimed in claim 2, wherein the molar ratio of sodium dihydrogen phosphate dihydrate, sodium hypochlorite and 2-methyl-2-butene in the step 2 is 4-10: 4-10, (3aR,10bR) -1-methylene-2-oxo-1, 3a,4,5,6,10 b-hexahydro-2H-furo [3,2-c ] pyrrolo [1,2-a ] azepine-8-carbaldehyde 1, 4-dioxane solution with the concentration of 0.01-1 mol/L and the volume of 1-5 m L.
6. The preparation method of the stemona alkaloid skeleton-based small molecule probe as claimed in claim 2, wherein the stirring temperature in step 2 is 28 ℃, the amount of glacial acetic acid is 2-5 m L, and the volume ratio of dichloromethane to methanol is 10: 1-20: 1.
7. The preparation method of the stemona alkaloid skeleton-based small molecule probe as claimed in claim 2, wherein the molar ratio of propargylamine, triethanolamine, 1-hydroxybenzotriazole and 1- (3-dimethylaminopropyl) -3-ethylcarbodiimide hydrochloride in the step 3 is 1.5-5: 10-15: 1.5-5: 5, (3aR,10bR) -1-methylene-2-oxo-1, 3a,4,5,6,10 b-hexahydro-2H-furo [3,2-c ] pyrrolo [1,2-a ] azepine-8-carboxylic acid in dichloromethane is 0.02-1 mol/L, the volume is 0.5-2 m L, and the number of extractions is 3 times.
8. The application of the small molecular probe based on the stemona alkaloid skeleton is characterized in that the small molecular probe can be used together with bioorthogonal reaction to perform action target fishing of a stemona alkaloid skeleton compound; can inhibit proliferation of colon cancer cell HCT-116.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202010264356.1A CN111423448B (en) | 2020-04-07 | 2020-04-07 | Stemona alkaloid skeleton-based small-molecule probe and preparation method and application thereof |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202010264356.1A CN111423448B (en) | 2020-04-07 | 2020-04-07 | Stemona alkaloid skeleton-based small-molecule probe and preparation method and application thereof |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN111423448A true CN111423448A (en) | 2020-07-17 |
| CN111423448B CN111423448B (en) | 2021-03-30 |
Family
ID=71557581
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN202010264356.1A Active CN111423448B (en) | 2020-04-07 | 2020-04-07 | Stemona alkaloid skeleton-based small-molecule probe and preparation method and application thereof |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN111423448B (en) |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN111297871A (en) * | 2020-04-07 | 2020-06-19 | 山西大学 | Application of radix stemonae alkaloid analogue |
Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN108864123A (en) * | 2018-07-11 | 2018-11-23 | 山西大学 | A kind of synthetic method of Stemona alkaloids |
| CN110016041A (en) * | 2019-05-13 | 2019-07-16 | 山西大学 | A kind of Stemona alkaloids analog and preparation method and purposes |
-
2020
- 2020-04-07 CN CN202010264356.1A patent/CN111423448B/en active Active
Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN108864123A (en) * | 2018-07-11 | 2018-11-23 | 山西大学 | A kind of synthetic method of Stemona alkaloids |
| CN110016041A (en) * | 2019-05-13 | 2019-07-16 | 山西大学 | A kind of Stemona alkaloids analog and preparation method and purposes |
Non-Patent Citations (2)
| Title |
|---|
| 姜登钊: "百部药材的生物碱类成分及生物活性研究进展", 《安徽农业科学》 * |
| 马爱军: "百部生物碱核心氮杂5,7并环骨架的合成", 《甘肃省化学会第二十八届年会暨第十届中学化学教学经验交流会论文集》 * |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN111297871A (en) * | 2020-04-07 | 2020-06-19 | 山西大学 | Application of radix stemonae alkaloid analogue |
Also Published As
| Publication number | Publication date |
|---|---|
| CN111423448B (en) | 2021-03-30 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| Yi et al. | Hydrogen bonding-mediated oligobenzamide foldamer receptors that efficiently bind a triol and saccharides in chloroform | |
| Lepage et al. | Synthesis of the first examples of iminosugar clusters based on cyclopeptoid cores | |
| CN104151301B (en) | A kind of fluorescent probe and its preparation method and application | |
| CN110684523A (en) | Near-infrared fluorescent molecular probe for detecting hydrogen sulfide and preparation method and application thereof | |
| CN109503553A (en) | A kind of light Affinity Probes molecule and preparation method thereof based on VEGFR-2 inhibitor B14 | |
| CN111423448A (en) | Stemona alkaloid skeleton-based small-molecule probe and preparation method and application thereof | |
| CN109456261A (en) | A kind of light Affinity Probes molecule and preparation method thereof based on VEGFR-2 inhibitor Sorafenib | |
| CN109369622A (en) | A kind of light Affinity Probes molecule and preparation method thereof based on VEGFR-2 inhibitor BD7 | |
| Tu et al. | A multi-component reaction for the synthesis of N-substituted furo [3, 4-b] quinoline derivatives under microwave irradiation | |
| CN107014993A (en) | The indirect competitive ELISA kit of cephalosporin analog antibiotic and its application in a kind of detection animal derived food | |
| Sallustrau et al. | Scalable and practical synthesis of clickable Cu-chelating azides | |
| Guo et al. | Spin‐labeled derivatives of cardiotonic steroids as tools for characterization of the extracellular entrance to the binding site on Na+, K+‐ATP ase | |
| Hirose et al. | Evaluation of the DNA Alkylation Properties of a Chlorambucil‐Conjugated Cyclic Pyrrole‐Imidazole Polyamide | |
| Cruces et al. | From phenylacetylphenylacetic acids to indoles: a simple new divergent synthesis of 6, 11-dihydro-5H-benzo [a] carbazol-5, 6-diones and 6, 11-dihydro-5H-benzo [b] carbazol-6, 11-diones | |
| CN112341467B (en) | Diketopiperazine alkaloid compound and preparation method and application thereof | |
| CN109879856B (en) | Multifunctional anti-Alzheimer disease benzimidazole derivative and preparation method and application thereof | |
| CN111808006B (en) | Nitrogen heterocyclic ring allyl compound and preparation method and application thereof | |
| CN113773283A (en) | Oxidobicycloheptene sulfonamide compound containing hydrophobic label and application thereof | |
| CN113912579A (en) | Rhodamine compound, preparation method thereof and application thereof in detection of microcystins | |
| CN111892588A (en) | Indole-benzothiazole derivative and preparation method and application thereof | |
| Yang et al. | Synthesis and Primary Biological Evaluation of Triazole‐Modified Picroside II Compounds | |
| CN114507201B (en) | Water wampee seed element derivative and preparation method and application thereof | |
| CN114409521B (en) | Compound extracted from symbiotic fungi Paraconiothyrium brasiliense and application thereof | |
| CN115160234B (en) | Aryl acetate-containing 1,4, 5-trisubstituted imidazole compound, preparation method and application | |
| CN115650981B (en) | Loganin-based monoterpene indole alkaloid, synthesis method and application thereof, and medicine for preventing and treating diabetes or diabetic nephropathy |
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 |