CN110437246A - The method that mesolow gradient silica gel dry column chromatography separates Neo-garcinolic acid, gambogicacid and the preparation of N- aryl gamboge amide - Google Patents
The method that mesolow gradient silica gel dry column chromatography separates Neo-garcinolic acid, gambogicacid and the preparation of N- aryl gamboge amide Download PDFInfo
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- CN110437246A CN110437246A CN201910763605.9A CN201910763605A CN110437246A CN 110437246 A CN110437246 A CN 110437246A CN 201910763605 A CN201910763605 A CN 201910763605A CN 110437246 A CN110437246 A CN 110437246A
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- silica gel
- gambogicacid
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- VYPSYNLAJGMNEJ-UHFFFAOYSA-N Silicium dioxide Chemical compound O=[Si]=O VYPSYNLAJGMNEJ-UHFFFAOYSA-N 0.000 title claims abstract description 125
- 239000000741 silica gel Substances 0.000 title claims abstract description 113
- 229910002027 silica gel Inorganic materials 0.000 title claims abstract description 113
- GEZHEQNLKAOMCA-RRZNCOCZSA-N (-)-gambogic acid Chemical compound C([C@@H]1[C@]2([C@@](C3=O)(C\C=C(\C)C(O)=O)OC1(C)C)O1)[C@H]3C=C2C(=O)C2=C1C(CC=C(C)C)=C1O[C@@](CCC=C(C)C)(C)C=CC1=C2O GEZHEQNLKAOMCA-RRZNCOCZSA-N 0.000 title claims abstract description 97
- GEZHEQNLKAOMCA-UHFFFAOYSA-N epiisogambogic acid Natural products O1C2(C(C3=O)(CC=C(C)C(O)=O)OC4(C)C)C4CC3C=C2C(=O)C2=C1C(CC=C(C)C)=C1OC(CCC=C(C)C)(C)C=CC1=C2O GEZHEQNLKAOMCA-UHFFFAOYSA-N 0.000 title claims abstract description 96
- GEZHEQNLKAOMCA-GXSDCXQCSA-N gambogic acid Natural products C([C@@H]1[C@]2([C@@](C3=O)(C\C=C(/C)C(O)=O)OC1(C)C)O1)[C@H]3C=C2C(=O)C2=C1C(CC=C(C)C)=C1O[C@@](CCC=C(C)C)(C)C=CC1=C2O GEZHEQNLKAOMCA-GXSDCXQCSA-N 0.000 title claims abstract description 96
- QALPNMQDVCOSMJ-UHFFFAOYSA-N isogambogic acid Natural products CC(=CCc1c2OC(C)(CC=C(C)C)C=Cc2c(O)c3C(=O)C4=CC5CC6C(C)(C)OC(CC=C(C)/C(=O)O)(C5=O)C46Oc13)C QALPNMQDVCOSMJ-UHFFFAOYSA-N 0.000 title claims abstract description 96
- 238000000034 method Methods 0.000 title claims abstract description 60
- 238000004440 column chromatography Methods 0.000 title claims abstract description 44
- 229940117709 gamboge Drugs 0.000 title claims abstract description 29
- 241000598860 Garcinia hanburyi Species 0.000 title claims abstract description 27
- 150000001408 amides Chemical class 0.000 title claims abstract description 22
- 238000002360 preparation method Methods 0.000 title abstract description 4
- 239000003480 eluent Substances 0.000 claims abstract description 58
- 238000000926 separation method Methods 0.000 claims abstract description 19
- 230000005526 G1 to G0 transition Effects 0.000 claims abstract description 12
- 150000004982 aromatic amines Chemical class 0.000 claims abstract description 10
- -1 1- ethyl Chemical group 0.000 claims abstract description 9
- 239000012043 crude product Substances 0.000 claims abstract description 7
- VHYFNPMBLIVWCW-UHFFFAOYSA-N 4-Dimethylaminopyridine Chemical compound CN(C)C1=CC=NC=C1 VHYFNPMBLIVWCW-UHFFFAOYSA-N 0.000 claims abstract description 4
- YMWUJEATGCHHMB-UHFFFAOYSA-N Dichloromethane Chemical compound ClCCl YMWUJEATGCHHMB-UHFFFAOYSA-N 0.000 claims description 46
- JUJWROOIHBZHMG-UHFFFAOYSA-N Pyridine Chemical compound C1=CC=NC=C1 JUJWROOIHBZHMG-UHFFFAOYSA-N 0.000 claims description 18
- CSCPPACGZOOCGX-UHFFFAOYSA-N Acetone Chemical compound CC(C)=O CSCPPACGZOOCGX-UHFFFAOYSA-N 0.000 claims description 17
- 238000006243 chemical reaction Methods 0.000 claims description 16
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 15
- 239000000284 extract Substances 0.000 claims description 14
- FPQQSJJWHUJYPU-UHFFFAOYSA-N 3-(dimethylamino)propyliminomethylidene-ethylazanium;chloride Chemical compound Cl.CCN=C=NCCCN(C)C FPQQSJJWHUJYPU-UHFFFAOYSA-N 0.000 claims description 13
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 claims description 10
- 238000001514 detection method Methods 0.000 claims description 9
- 239000012452 mother liquor Substances 0.000 claims description 9
- UMJSCPRVCHMLSP-UHFFFAOYSA-N pyridine Natural products COC1=CC=CN=C1 UMJSCPRVCHMLSP-UHFFFAOYSA-N 0.000 claims description 9
- 238000005406 washing Methods 0.000 claims description 9
- 230000006837 decompression Effects 0.000 claims description 8
- 239000000706 filtrate Substances 0.000 claims description 8
- NOKXCPFWTMTBTO-UHFFFAOYSA-N 3-(iminomethylideneamino)-n,n-dimethylpentan-1-amine;hydrochloride Chemical compound Cl.N=C=NC(CC)CCN(C)C NOKXCPFWTMTBTO-UHFFFAOYSA-N 0.000 claims description 6
- CSNNHWWHGAXBCP-UHFFFAOYSA-L Magnesium sulfate Chemical compound [Mg+2].[O-][S+2]([O-])([O-])[O-] CSNNHWWHGAXBCP-UHFFFAOYSA-L 0.000 claims description 6
- 239000004615 ingredient Substances 0.000 claims description 6
- 239000012074 organic phase Substances 0.000 claims description 6
- 239000002253 acid Substances 0.000 claims description 5
- 238000002156 mixing Methods 0.000 claims description 5
- 239000012044 organic layer Substances 0.000 claims description 5
- 238000010791 quenching Methods 0.000 claims description 5
- 230000000171 quenching effect Effects 0.000 claims description 5
- 238000001228 spectrum Methods 0.000 claims description 5
- 238000003756 stirring Methods 0.000 claims description 5
- AULWPXHFRBLPAE-UHFFFAOYSA-N 6-chloropyridine Chemical compound ClC1=C=CC=C[N]1 AULWPXHFRBLPAE-UHFFFAOYSA-N 0.000 claims description 4
- QGZKDVFQNNGYKY-UHFFFAOYSA-N Ammonia Chemical compound N QGZKDVFQNNGYKY-UHFFFAOYSA-N 0.000 claims description 4
- UGAPHEBNTGUMBB-UHFFFAOYSA-N acetic acid;ethyl acetate Chemical compound CC(O)=O.CCOC(C)=O UGAPHEBNTGUMBB-UHFFFAOYSA-N 0.000 claims description 4
- 238000004587 chromatography analysis Methods 0.000 claims description 4
- 238000010828 elution Methods 0.000 claims description 4
- 239000002024 ethyl acetate extract Substances 0.000 claims description 4
- 150000001412 amines Chemical class 0.000 claims description 3
- 238000001035 drying Methods 0.000 claims description 3
- 239000000843 powder Substances 0.000 claims description 3
- 229910021529 ammonia Inorganic materials 0.000 claims description 2
- 238000010438 heat treatment Methods 0.000 claims description 2
- 239000012535 impurity Substances 0.000 claims description 2
- 230000020477 pH reduction Effects 0.000 claims description 2
- 230000036961 partial effect Effects 0.000 claims description 2
- LMDZBCPBFSXMTL-UHFFFAOYSA-N 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide Chemical compound CCN=C=NCCCN(C)C LMDZBCPBFSXMTL-UHFFFAOYSA-N 0.000 claims 1
- QJITUZDKRABEGK-UHFFFAOYSA-N n'-propylmethanediimine;hydrochloride Chemical compound Cl.CCCN=C=N QJITUZDKRABEGK-UHFFFAOYSA-N 0.000 claims 1
- QOSSAOTZNIDXMA-UHFFFAOYSA-N Dicylcohexylcarbodiimide Chemical compound C1CCCCC1N=C=NC1CCCCC1 QOSSAOTZNIDXMA-UHFFFAOYSA-N 0.000 abstract description 13
- 238000006555 catalytic reaction Methods 0.000 abstract description 9
- 229910052739 hydrogen Inorganic materials 0.000 abstract description 3
- 238000011938 amidation process Methods 0.000 abstract 1
- DCPMPXBYPZGNDC-UHFFFAOYSA-N hydron;methanediimine;chloride Chemical compound Cl.N=C=N DCPMPXBYPZGNDC-UHFFFAOYSA-N 0.000 abstract 1
- 229960001866 silicon dioxide Drugs 0.000 description 89
- 239000000523 sample Substances 0.000 description 21
- 239000004576 sand Substances 0.000 description 7
- 239000002904 solvent Substances 0.000 description 7
- 241000251468 Actinopterygii Species 0.000 description 6
- 239000002245 particle Substances 0.000 description 6
- XEKOWRVHYACXOJ-UHFFFAOYSA-N Ethyl acetate Chemical compound CCOC(C)=O XEKOWRVHYACXOJ-UHFFFAOYSA-N 0.000 description 5
- VEXZGXHMUGYJMC-UHFFFAOYSA-N Hydrochloric acid Chemical compound Cl VEXZGXHMUGYJMC-UHFFFAOYSA-N 0.000 description 5
- ZMANZCXQSJIPKH-UHFFFAOYSA-N Triethylamine Chemical compound CCN(CC)CC ZMANZCXQSJIPKH-UHFFFAOYSA-N 0.000 description 5
- 238000010521 absorption reaction Methods 0.000 description 5
- QVGXLLKOCUKJST-UHFFFAOYSA-N atomic oxygen Chemical compound [O] QVGXLLKOCUKJST-UHFFFAOYSA-N 0.000 description 5
- 238000009395 breeding Methods 0.000 description 5
- 230000001488 breeding effect Effects 0.000 description 5
- 239000003814 drug Substances 0.000 description 5
- 239000001301 oxygen Substances 0.000 description 5
- 229910052760 oxygen Inorganic materials 0.000 description 5
- 239000012071 phase Substances 0.000 description 5
- 238000001179 sorption measurement Methods 0.000 description 5
- 206010028980 Neoplasm Diseases 0.000 description 4
- 235000019441 ethanol Nutrition 0.000 description 4
- 239000010410 layer Substances 0.000 description 4
- 239000007788 liquid Substances 0.000 description 4
- 239000000203 mixture Substances 0.000 description 4
- 238000011084 recovery Methods 0.000 description 4
- 238000009738 saturating Methods 0.000 description 4
- OKKJLVBELUTLKV-UHFFFAOYSA-N Methanol Chemical compound OC OKKJLVBELUTLKV-UHFFFAOYSA-N 0.000 description 3
- 230000002411 adverse Effects 0.000 description 3
- 230000015572 biosynthetic process Effects 0.000 description 3
- 230000000052 comparative effect Effects 0.000 description 3
- 239000002274 desiccant Substances 0.000 description 3
- XHIKCWBOQBWVPU-UHFFFAOYSA-N dichloromethane;n,n-diethylethanamine;ethanol Chemical compound CCO.ClCCl.CCN(CC)CC XHIKCWBOQBWVPU-UHFFFAOYSA-N 0.000 description 3
- 238000011068 loading method Methods 0.000 description 3
- 230000014759 maintenance of location Effects 0.000 description 3
- 230000008569 process Effects 0.000 description 3
- 230000002829 reductive effect Effects 0.000 description 3
- 239000000377 silicon dioxide Substances 0.000 description 3
- 241000345998 Calamus manan Species 0.000 description 2
- 241000598812 Garcinia tinctoria Species 0.000 description 2
- 206010058467 Lung neoplasm malignant Diseases 0.000 description 2
- KAESVJOAVNADME-UHFFFAOYSA-N Pyrrole Chemical compound C=1C=CNC=1 KAESVJOAVNADME-UHFFFAOYSA-N 0.000 description 2
- 208000005718 Stomach Neoplasms Diseases 0.000 description 2
- DTQVDTLACAAQTR-UHFFFAOYSA-N Trifluoroacetic acid Chemical compound OC(=O)C(F)(F)F DTQVDTLACAAQTR-UHFFFAOYSA-N 0.000 description 2
- 230000009435 amidation Effects 0.000 description 2
- 238000007112 amidation reaction Methods 0.000 description 2
- 230000000259 anti-tumor effect Effects 0.000 description 2
- 230000008901 benefit Effects 0.000 description 2
- 210000004204 blood vessel Anatomy 0.000 description 2
- 201000011510 cancer Diseases 0.000 description 2
- 239000003795 chemical substances by application Substances 0.000 description 2
- 238000003795 desorption Methods 0.000 description 2
- 238000004090 dissolution Methods 0.000 description 2
- 230000000694 effects Effects 0.000 description 2
- 238000005516 engineering process Methods 0.000 description 2
- 238000011049 filling Methods 0.000 description 2
- 206010017758 gastric cancer Diseases 0.000 description 2
- 239000011521 glass Substances 0.000 description 2
- 238000004128 high performance liquid chromatography Methods 0.000 description 2
- 230000007794 irritation Effects 0.000 description 2
- 201000005202 lung cancer Diseases 0.000 description 2
- 208000020816 lung neoplasm Diseases 0.000 description 2
- 238000004519 manufacturing process Methods 0.000 description 2
- 230000004048 modification Effects 0.000 description 2
- 238000012986 modification Methods 0.000 description 2
- VLKZOEOYAKHREP-UHFFFAOYSA-N n-Hexane Chemical compound CCCCCC VLKZOEOYAKHREP-UHFFFAOYSA-N 0.000 description 2
- 229930014626 natural product Natural products 0.000 description 2
- 238000006213 oxygenation reaction Methods 0.000 description 2
- 125000001997 phenyl group Chemical group [H]C1=C([H])C([H])=C(*)C([H])=C1[H] 0.000 description 2
- 235000012950 rattan cane Nutrition 0.000 description 2
- 230000036632 reaction speed Effects 0.000 description 2
- 238000012827 research and development Methods 0.000 description 2
- 238000002390 rotary evaporation Methods 0.000 description 2
- 125000005372 silanol group Chemical group 0.000 description 2
- 238000010898 silica gel chromatography Methods 0.000 description 2
- 201000011549 stomach cancer Diseases 0.000 description 2
- 238000003786 synthesis reaction Methods 0.000 description 2
- 238000012360 testing method Methods 0.000 description 2
- 238000005303 weighing Methods 0.000 description 2
- COVMVPHACFXMAX-OYNOKLRGSA-N (-)-morellic acid Chemical compound C1=CC(C)(C)OC2=C1C(O)=C1C(=O)C3=C[C@@H](C(=O)[C@]4(C\C=C(\C)C(O)=O)OC5(C)C)C[C@@H]5[C@]34OC1=C2CC=C(C)C COVMVPHACFXMAX-OYNOKLRGSA-N 0.000 description 1
- ADFXKUOMJKEIND-UHFFFAOYSA-N 1,3-dicyclohexylurea Chemical compound C1CCCCC1NC(=O)NC1CCCCC1 ADFXKUOMJKEIND-UHFFFAOYSA-N 0.000 description 1
- 206010006187 Breast cancer Diseases 0.000 description 1
- 208000026310 Breast neoplasm Diseases 0.000 description 1
- VDSCKSOYNLTQSY-UHFFFAOYSA-N Garcinolic acid Chemical class O1C2(C(OC3(C)C)(CC=C(C)C(O)=O)C(O)=O)C3CCC=C2C(=O)C2=C1C(CC=C(C)C)=C1OC(CCC=C(C)C)(C)C=CC1=C2O VDSCKSOYNLTQSY-UHFFFAOYSA-N 0.000 description 1
- 206010027476 Metastases Diseases 0.000 description 1
- XNIZIBPYBUCVEU-UHFFFAOYSA-N Morellic acid Natural products CC1Oc2c(CC=C(C)C)c3OC45C6CC(C=C4C(=O)c3c(O)c2C=C1)C(=O)C5(CC=C(C)/C(=O)O)OC6(C)C XNIZIBPYBUCVEU-UHFFFAOYSA-N 0.000 description 1
- XUIMIQQOPSSXEZ-UHFFFAOYSA-N Silicon Chemical group [Si] XUIMIQQOPSSXEZ-UHFFFAOYSA-N 0.000 description 1
- 238000005411 Van der Waals force Methods 0.000 description 1
- 238000004458 analytical method Methods 0.000 description 1
- 230000033115 angiogenesis Effects 0.000 description 1
- RJGDLRCDCYRQOQ-UHFFFAOYSA-N anthrone Chemical compound C1=CC=C2C(=O)C3=CC=CC=C3CC2=C1 RJGDLRCDCYRQOQ-UHFFFAOYSA-N 0.000 description 1
- 230000006907 apoptotic process Effects 0.000 description 1
- 125000003118 aryl group Chemical group 0.000 description 1
- 230000000975 bioactive effect Effects 0.000 description 1
- 239000006227 byproduct Substances 0.000 description 1
- 125000002915 carbonyl group Chemical group [*:2]C([*:1])=O 0.000 description 1
- 125000003178 carboxy group Chemical group [H]OC(*)=O 0.000 description 1
- 150000007942 carboxylates Chemical group 0.000 description 1
- 210000004027 cell Anatomy 0.000 description 1
- 230000022131 cell cycle Effects 0.000 description 1
- 230000004663 cell proliferation Effects 0.000 description 1
- 230000008859 change Effects 0.000 description 1
- 238000012512 characterization method Methods 0.000 description 1
- 239000007795 chemical reaction product Substances 0.000 description 1
- 239000003153 chemical reaction reagent Substances 0.000 description 1
- 239000000084 colloidal system Substances 0.000 description 1
- 150000001875 compounds Chemical class 0.000 description 1
- 230000007812 deficiency Effects 0.000 description 1
- 238000013461 design Methods 0.000 description 1
- 238000009792 diffusion process Methods 0.000 description 1
- 238000009509 drug development Methods 0.000 description 1
- 230000007613 environmental effect Effects 0.000 description 1
- QWKLHPQCHMOZAP-UHFFFAOYSA-N ethyl acetate;n-ethylethanamine;methanol Chemical compound OC.CCNCC.CCOC(C)=O QWKLHPQCHMOZAP-UHFFFAOYSA-N 0.000 description 1
- FPVGTPBMTFTMRT-NSKUCRDLSA-L fast yellow Chemical compound [Na+].[Na+].C1=C(S([O-])(=O)=O)C(N)=CC=C1\N=N\C1=CC=C(S([O-])(=O)=O)C=C1 FPVGTPBMTFTMRT-NSKUCRDLSA-L 0.000 description 1
- 238000001914 filtration Methods 0.000 description 1
- 239000008236 heating water Substances 0.000 description 1
- 210000003958 hematopoietic stem cell Anatomy 0.000 description 1
- 239000001257 hydrogen Substances 0.000 description 1
- 125000002887 hydroxy group Chemical group [H]O* 0.000 description 1
- 238000001727 in vivo Methods 0.000 description 1
- 230000002401 inhibitory effect Effects 0.000 description 1
- 230000005764 inhibitory process Effects 0.000 description 1
- 238000007689 inspection Methods 0.000 description 1
- 238000002955 isolation Methods 0.000 description 1
- COVMVPHACFXMAX-ZVKSWBPMSA-N isomorellic acid Natural products O=C(O)/C(=C\C[C@]12C(=O)[C@H]3C=C4C(=O)c5c(O)c6c(c(C/C=C(\C)/C)c5O[C@]14[C@@H](C(C)(C)O2)C3)OC(C)(C)C=C6)/C COVMVPHACFXMAX-ZVKSWBPMSA-N 0.000 description 1
- 238000011031 large-scale manufacturing process Methods 0.000 description 1
- 150000002611 lead compounds Chemical class 0.000 description 1
- 238000004811 liquid chromatography Methods 0.000 description 1
- 210000004185 liver Anatomy 0.000 description 1
- 201000007270 liver cancer Diseases 0.000 description 1
- 208000014018 liver neoplasm Diseases 0.000 description 1
- 230000007246 mechanism Effects 0.000 description 1
- 238000005272 metallurgy Methods 0.000 description 1
- 238000005065 mining Methods 0.000 description 1
- 238000012544 monitoring process Methods 0.000 description 1
- 239000002547 new drug Substances 0.000 description 1
- 239000003921 oil Substances 0.000 description 1
- 210000000496 pancreas Anatomy 0.000 description 1
- 238000005192 partition Methods 0.000 description 1
- 238000012805 post-processing Methods 0.000 description 1
- 125000002924 primary amino group Chemical group [H]N([H])* 0.000 description 1
- 239000000047 product Substances 0.000 description 1
- 230000035755 proliferation Effects 0.000 description 1
- 238000005086 pumping Methods 0.000 description 1
- 239000002994 raw material Substances 0.000 description 1
- 239000013558 reference substance Substances 0.000 description 1
- 239000011347 resin Substances 0.000 description 1
- 229920005989 resin Polymers 0.000 description 1
- 239000012488 sample solution Substances 0.000 description 1
- 238000005070 sampling Methods 0.000 description 1
- 238000007086 side reaction Methods 0.000 description 1
- 229910052710 silicon Inorganic materials 0.000 description 1
- 150000003384 small molecules Chemical class 0.000 description 1
- 230000003595 spectral effect Effects 0.000 description 1
- 230000001988 toxicity Effects 0.000 description 1
- 231100000419 toxicity Toxicity 0.000 description 1
- 238000012546 transfer Methods 0.000 description 1
- 210000004881 tumor cell Anatomy 0.000 description 1
- 238000002137 ultrasound extraction Methods 0.000 description 1
- 239000002699 waste material Substances 0.000 description 1
- 150000007964 xanthones Chemical class 0.000 description 1
Classifications
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07D—HETEROCYCLIC COMPOUNDS
- C07D493/00—Heterocyclic compounds containing oxygen atoms as the only ring hetero atoms in the condensed system
- C07D493/02—Heterocyclic compounds containing oxygen atoms as the only ring hetero atoms in the condensed system in which the condensed system contains two hetero rings
- C07D493/10—Spiro-condensed systems
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07D—HETEROCYCLIC COMPOUNDS
- C07D493/00—Heterocyclic compounds containing oxygen atoms as the only ring hetero atoms in the condensed system
- C07D493/12—Heterocyclic compounds containing oxygen atoms as the only ring hetero atoms in the condensed system in which the condensed system contains three hetero rings
- C07D493/20—Spiro-condensed systems
Landscapes
- Chemical & Material Sciences (AREA)
- Organic Chemistry (AREA)
- Organic Low-Molecular-Weight Compounds And Preparation Thereof (AREA)
- Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
Abstract
The present invention relates to mesolow gradient silica gel dry column chromatography separation Neo-garcinolic acid, gambogicacid and the methods of N- aryl gamboge amide preparation, make stationary phase using selected eluant, eluent with silica gel H, GA the and GNA sample that purity is respectively 90%, 93% can be isolated inexpensive, easyly, fairly large through mesolow gradient silica gel dry column chromatography；It is stirred at room temperature down, the amidation process of 1- ethyl -3- (3-N, N- dimethylamino-propyl) carbodiimide hydrochloride (or dicyclohexylcarbodiimide) and 4- (N, N- dimethylamino) pyridine catalysis gambogicacid and substituted aromatic amines.Crude product makees the silica gel medium pressure dry column chromatography of stationary phase through silica gel H, can prepare the N- aryl gamboge amide sample of high-purity inexpensive, easyly, fairly large.
Description
Technical field
Extracting and developing and characterization bioactive small molecule, are that lead compound is obtained in new drug development from natural products
One of main path.The present invention relates to mesolow gradient silica gel dry column chromatography separation Neo-garcinolic acid, gambogicacid and preparation N-
The new method of aryl gamboge amide, is belonging respectively to Separation of Natural Products and technical field of medicine synthesis.
Background technique
Effective component gambogicacid (GA) in Chinese medicine gamboge is to tumour cells such as liver cancer, lung cancer, gastric cancer, rhinocarcinoma and cancers of pancreas
Strain proliferation has apparent inhibiting effect.Under the premise of having little influence on normal hematopoietic cell and leucocyte, GA can lead to
Inhibition cell Proliferation is crossed, arresting cell cycle, induces cell apoptosis, inhibit the mechanism such as metastases and angiogenesis selectively
Kill cancer cell.Another effective component Neo-garcinolic acid (GNA) in gamboge, body is inside and outside also to inhibit to more efficient lung cancer, liver
The growth of the kinds of tumors such as cancer, gastric cancer, breast cancer.GA, GNA are expected to be developed into an antitumor kind new medicine.But in gamboge
GNA content is lower, and coexists to the ingredient of multiple similar structures.Existing GNA separation method, cannot be easy, inexpensive, efficient
The GNA of high-purity is isolated to rate from the Resina garciniae extract containing other ingredients such as GA.
In addition the disadvantages of strong blood vessel irritation, low aqueous solubility, the Half-life in vivo of GA, GNA be short, low activity, poor stability,
And limit the bottleneck of such medicament research and development.It is introduced in the molecule by the amide that the converting carboxylate groups in GA, GNA are replaced at N-
Hydrophilic radical and pharmacophore, it is expected to enhance the bioactivity of GA, GNA class entity, improve it is water-soluble, reduce blood vessel irritation,
Improve intracorporal pharmacokinetics process of rat etc..As can simply, efficiently isolating GA, GNA, then solving limitation should
The bottleneck problem of class medicament research and development.
The method that GNA separation at present is mostly separated using multiple silica gel column chromatography, macroporous resin column chromatography.But both sides
Method has that serious dead absorption, time-consuming, organic reagent is using more and complex steps and the problems such as increase separation costs.Recklessly
Clock etc. uses thick silica gel to be stationary phase, separate GNA by eluant, eluent column chromatography of methanol-ethyl acetate-diethylamine solvent combination,
Not only solvent toxicity is big and inferior separating effect (design of Hu Zhong Novel garcinolic acid derivative, synthesis and anti-tumor activity are tentatively ground
Study carefully Anhui University of Chinese Medecine M Sc thesis, 2013).Preparative high-performance liquid chromatographic method separate GNA when applied sample amount it is small,
It is at high cost, and limit the preparative-scale of GNA sample.Chinese patent literature CN105061455A discloses a kind of high-speed counter-current color
Spectral technology can simultaneously massively separating high purity gambogicacid and Neo-garcinolic acid method, which comprises use n-hexane, acetic acid
Ethyl ester, methanol, water is by volume (6.5-7.5): (2.5-3.5): (6.5-8.5): after (1.5-3.5) mixing, standing is divided into,
Lower two-phase；Using it is suitable it is described on mix solvent dissolution gamboge alcohol extracts, obtain sample solution.Respectively to contain
The upper phase of trifluoroacetic acid, the lower phase containing triethylamine are stationary phase and mobile phase, using high-speed counter-current chromatograph separation, ultraviolet inspection
It surveys device monitoring and collects concentration, drying after target fraction respectively.But this high speed adverse current chromatogram method is due to every chromatograph price
Ten thousand RMB of 50-60, equipment cost is high, limits the universal of the isolation technics.In addition there are no N- aryl gamboge amides so far
The document report of aspect is prepared, therefore there is an urgent need to find gamboge extracting solution to be isolated major part easyly, fairly large
Remnants GNA is separated and the amidated method of GA virtue with ingredients such as GA in the mother liquor of gambogicacid pyridiniujm.For this purpose, special propose this hair
It is bright.
Summary of the invention
In view of the deficiencies of the prior art, it is an object of the present invention to provide one kind filters out gambogicacid from Resina garciniae extract
The method of high-purity GA, GNA is isolated in the mother liquor of pyridiniujm.The present invention uses mesolow gradient silica gel dry column chromatography technology,
Sample to be separated is mounted in the upper end of chromatographic grade silica gel H, eluant, eluent introduces from the upper end of sample and presses flowing side from top to bottom
To gradually by being stringed out in silica gel by isolated compound by the sequence of polarity size in addition to GA.It is low in use of the present invention
Gradient silica gel dry column chromatography is pressed, simply, conveniently, is suitble to large-scale production.
Another object of the present invention is to provide using the GA isolated as primary raw material, the side of N- aryl gamboge amide is prepared
Method.
Term explanation:
Gambogicacid (GA): molecular formula C38H44O8, No. CAS: 2752-65-0, structural formula is as follows:
Neo-garcinolic acid (GNA): molecular formula: C38H46O9, No. CAS: 93772-31-7, structural formula is as follows:
Rf' value=GNA spot centers are to parallax range/GA spot centers to parallax range.
Acetic acid ethyl acetate extract after the mother liquor acidification of gambogicacid pyridiniujm of the present invention, obtains as follows:
The pyrrole that volume is 1.5 times of medicinal extract weight is added after the medicinal extract weighing that goa powder is concentrated to get with acetone ultrasonic extraction liquid
40 DEG C of warm water of 0.13 times of filtrate volume successively room after mixing evenly is largely added after dissolution for pyridine, 70 DEG C of heating water baths in filtrate
It is stood in temperature, refrigerator and gambogicacid pyridiniujm is precipitated.Be concentrated under reduced pressure in 70 DEG C of filtrate rotary evaporators it is close dry, take 1g residue with
After the mixing of 5mL water plus 1% dilute hydrochloric acid tune pH=4, ethyl acetate (4mL × 3) extract.Combined extract washes (2mL × 2) into
Property after, it is dry to separate organic phase anhydrous magnesium sulfate.
According to the above method in gamboge acid mother liquor obtained in addition to mainly containing gambogicacid, Neo-garcinolic acid, also containing another oxygen
Miscellaneous anthrone bridged ring (xanthone) class main component gambogicacid B (morellic acid).
Technical scheme is as follows:
The method that mesolow silica gel dry column chromatography separates Neo-garcinolic acid, gambogicacid, comprises the following steps that
The mother liquor acid of gambogicacid pyridiniujm to be precipitated after the medicinal extract of goa powder acetone extract successively pyridine, water temperature heat treatment
Acetic acid ethyl acetate extract after change is sample to be separated, makees stationary phase, with silica gel H respectively with dichloro-ethane-ethanol-triethylamine three
First system is eluant, eluent, is separated using the flow direction of mesolow gradient silica gel H dry column chromatography eluant, eluent from top to bottom, is made new
Gambogicacid is separated into different bands of a spectrum from ingredients such as gambogicacid B, impurity on silica gel H column, and gambogicacid is then by from silica gel H column
Gambogicacid eluent is all eluted and collects, ethanol washing silica gel H is simultaneously concentrated to get Neo-garcinolic acid；The concentration of gambogicacid eluent,
It is i.e. another to obtain gambogicacid.
, according to the invention it is preferred to, gambogicacid is all eluted from silica gel H column, collects gambogicacid using clean tube
Eluent.TLC method and GA reference substance can be used and be total to thin layer detection with plate.
Dry method loading is carried out when mesolow gradient silica gel dry column chromatography separates GA, GNA in the present invention: need to be to filtering out gamboge
Residue after the mother liquor of sour pyridiniujm is concentrated under reduced pressure successively uses the acetic acid ethyl acetate extracts such as 1% hydrochloric acid tune pH 3-4, GA and GNA
Concentration uses acetone solution as few as possible after closely doing, and thick silica gel as few as possible is added just to cover the acetone solns such as GA, GNA
Careful rotary evaporation removes acetone (general crude product, which accounts for, mixes the 50- of sample total weight to get high-content as far as possible again for upper surface
60%), gamboge acid crude-silica-gel mixture to be separated is used as column and chromatographs dry method loading.
, according to the invention it is preferred to, eluant, eluent is divided into eluant, eluent 1 and eluant, eluent 2, dichloroethanes-second in eluant, eluent 1
Alcohol-triethylamine volume ratio is 30:0.15:4, and dichloro-ethane-ethanol-triethylamine volume ratio is 20:0.6:3 in eluant, eluent 2.
First with eluant, eluent 1 elution until 1 forward position of eluant, eluent ran silica gel H bottom and receive may the eluant, eluent 1 containing GA, reuse
Eluant, eluent 2 is eluted and is received, until the detection of silica gel tlc method receives GA in fraction and is all eluted in gambogicacid eluent.
, according to the invention it is preferred to, silica gel H and example weight to be separated ratio are 60-90:1.
, according to the invention it is preferred to, silica gel H height and chromatography column internal diameter ratio are greater than 8:1.
, according to the invention it is preferred to, eluant, eluent fair current is carried out by 3-4cm height/min or 1 drop/s speed.
, according to the invention it is preferred to, partial size < 38 μm of silica gel H.
According to the present invention, the side of high-purity GNA, GA is isolated from the mother liquor that Resina garciniae extract filters out gambogicacid pyridiniujm
Method, a kind of preferred embodiment, comprising the following steps:
(1) by from top to bottom in upper end ground, the vertical placement glass column for having glass piston with saturating sand plate, lower end
Sequence, be sequentially filled silica gel H, 200-300 mesh mixes water pump after the dry silica gel of sample, fine sand thin layer and takes out real, turn off chromatographic column lower end
Piston and termination of pumping；
(2) dress eluant, eluent constant pressure funnel piston, bulbs,double or oxygen pump of breeding fish pressurization, chromatographic column lower end are successively opened
Piston makes eluant, eluent dry column chromatography until the bottom of silica gel H was run in enough 1 forward positions of eluant, eluent and clean tube reception is a certain amount of
Eluant, eluent, then use instead eluant, eluent 2 to silica gel tlc detect receive fraction in GA be all eluted in 3-5 clean tube, its
Its effective component separates several colour bands on silica gel H；
(3) bulbs,double is successively removed or oxygenation of breeding fish after the eluant, eluent of silica gel H upper end just submerges silica gel H upper surface
Pump, constant pressure funnel, blowout silica gel, will be in silica gel H and in TLC method into clean pallet after chromatographic column bottom end connects inflator
GNA Rf' the corresponding bands of a spectrum of value are uniformly divided into 4-6 sections, and every section takes silica gel H sample TLC method detection wherein GNA purity；
(4) merge above sterling containing GNA, the silica gel H compared with sterling respectively, 95% ethyl alcohol is cleaned GNA in each section of silica gel H, washed
Solution decompression rotary evaporation after washing is concentrated to get GNA；
(5) merge the reception fraction containing pure GA in step (2) test tube and depressurize rotary evaporated to dryness to get GA.
In the present invention mesolow gradient silica gel H dry column chromatography mainly by between silica gel particle hole generate capillarity,
Eluant, eluent liquid level and eluant, eluent liquid level difference, bulbs,double (or oxygen pump of breeding fish) in the dry column of silica gel H in constant pressure funnel
The pressure of generation, and each ingredient in sample to be tested is unfolded along the moving direction of solvent.Silica Surface removes and component to be separated
It is intermolecular there are outside Van der Waals force, the silanol group on its surface can also be with the poles such as carbonyl, carboxyl, hydroxyl in component molecular to be separated
There is hydrogen bond actions for property group.In column chromatography when the mixed component in Resina garciniae extract passes through silicagel column, the knot such as GA, GNA
Difference on structure but also they from silica gel H to be tightly combined degree different.In this way through Adsorption and desorption many times it is attached, inhale again
Attached, desorption again, so that the moving distance on silica gel H such as GA, GNA is different.With the common column color of granularity 200-300 purpose
Spectrum silica gel (45-75 μm of particle diameter) is compared, because of eluant, eluent and sample during silica gel H column chromatography exhibition layer of the diameter less than 38 μm
Molecule silica gel particle deep hole more difficult to get access, so that GA, GNA adsorption and de-adsorption process occur mainly on the outer surface of particle.
Thus overcome the molecules such as GA, GNA due to pass in and out silica gel particle deep hole, caused by diffusion and mass transfer it is slow.With silica gel H bottom end
The countercurrent column chromatography for adding sample to be separated, eluant, eluent to move from the bottom up is compared, this mesolow gradient silica gel H dry column chromatography pair
The separation of sample has the advantages that save silica gel, energy Gradient chromatography.
Another aspect of the present invention, provides the GA that the above method the is isolated method for preparing N- aryl gamboge amide, including with
Lower step:
(1) by GA, 1- ethyl -3- (N, N- dimethylamino) propyl carbodiimide hydrochloride (EDCI.HCl), 4- (N, N- bis-
Methylamino) pyridine (DMAP) and methylene chloride (DCM) mixing, the dichloromethane solution that arylamine is added after 15min, room is stirred at room temperature
Temperature continues stirring 6-12h GA spot into silica gel tlc method detection reaction solution and completely disappears；
(2) plus after water quenching reaction organic layer is separated, the drying of organic phase anhydrous magnesium sulfate, filtrate decompression warp is separated after washing
After rotary evaporator concentration, crude product makees the mesolow gradient dry column chromatography of stationary phase through silica gel H to get the N- aryl of high-purity
Gamboge amide sample.
, according to the invention it is preferred to, GA, 1- ethyl -3- (N, N- dimethylamino) propyl carbodiimide hydrochloride
(EDCI.HCl) molar ratio of (or dicyclohexylcarbodiimide, DCC) and 4- (N, N- dimethylamino) pyridine (DMAP) is 1:
(1.05-1.15): (0.2-0.4)；
Preferably, the molar ratio of GA and arylamine is 1:(1.1-1.2).
, according to the invention it is preferred to, the arylamine is the chloro- 3- 5-trifluoromethylaniline of 4- or 3- (6- chloropyridine base) amine.
It is as follows that the present invention prepares N- aryl gamboge amide reaction equation:
The invention has the benefit that
1, separation method of the present invention is using in dichloro-ethane-ethanol-triethylamine solvent group cooperation eluant, eluent of hypotoxicity
The separation of low pressure gradient dry column chromatography reduces the three wastes, is conducive to environmental protection；Applied sample amount is much larger than the limit for being typically prepared chromatography,
Strong support is provided for subsequent industrialized production；Loading is easy to implement the method, time saving, color needed for sample intercepts after separating in column
Section is eluted and is concentrated.This method ensure that the stability that chromatographic column efficiently separates, method is easy, efficiently.
2, with 1- ethyl -3- (N, N- dimethylamino) propyl carbodiimide hydrochloride (EDCI.HCl) and 4- (N, N- diformazan
Amino) pyridine (DMAP) joint catalysis the amidation of GA virtue compare, dicyclohexylcarbodiimide (DCC) and 4- (N, N- diformazan ammonia
Base) pyridine (DMAP) though joint catalysis it is at low cost but the easy moisture absorption, reaction speed are slow, and product is not readily separated.
Detailed description of the invention
Fig. 1 is the schematic device that GA, GNA are separated in the embodiment of the present invention 1,
Fig. 2 is the process flow chart for separating GA, GNA.
Specific embodiment
Below in conjunction with specific embodiment of the present invention, technical solution of the present invention is clearly and completely described.Obviously
Described embodiment is only a part of the embodiments of the present invention, instead of all the embodiments.Based on the reality in the present invention
Example is applied, every other embodiment obtained by those of ordinary skill in the art without making creative efforts all belongs to
In the scope of protection of the invention.
The schematic device of GA, GNA are separated used in embodiment, as shown in Figure 1.The device include bottom end band piston,
The constant pressure funnel with piston, and constant pressure funnel are installed in the chromatographic column with saturating sand plate of upper end ground, chromatographic column upper end
Upper end connects bulbs,double or oxygen increasing pump of breeding fish.Chromatographic column bottom end will connect water pump when filling column.
Embodiment 1
The present invention passes through separator shown in FIG. 1 (chromatographic column is equipped with silica gel H) selection dichloro-ethane-ethanol-triethylamine
System is the disposable mesolow gradient silica gel H dry column chromatography of eluant, eluent, and gambogicacid pyridiniujm can be filtered out from Resina garciniae extract
GA, GNA of high-purity are isolated in mother liquor.
The specific process steps are as follows:
(1) bottom end as shown in Figure 1 with piston, upper end ground the chromatographic column with saturating sand plate in be sequentially filled 110 DEG C of work
Changing the silica gel H of 2h, [silica gel height: internal diameter > 8:1 and example weight to be separated ratio (60-90): 1], to mix the thick silica gel of sample, 2-3mm thin
After sand bed, installs and taken out in fact with piston, the constant pressure funnel for filling eluant, eluent, water pump；
(2) bottom end piston, the air extractor of chromatographic column are successively closed, then successively opens dress eluant, eluent constant pressure funnel
Bottom piston, breed fish oxygen pump or the ball pressurization of two companies, the appropriate piston for opening chromatographic column bottom end, by 3-4cm silica gel length/min
Or the speed of 1-2d/s start in press dry column chromatography until 1 forward position of eluant, eluent ran the saturating sand plate of silica gel H the lowermost and receive one
Quantitative eluant, eluent, then use the GA into silica gel tlc method detection reception fraction of eluant, eluent 2 instead and be all eluted to 3-5 clean examination
Guan Zhong, and other samples several colour bands are separated on silica gel H after stop column chromatography；
(3) bulbs,double (or oxygenation of breeding fish successively is removed after the eluant, eluent 2 of silica gel upper end just submerges fine sand upper surface
Pump), constant pressure funnel, chromatographic column bottom end connect inflator after blowout silica gel into clean pallet, will be with rattan new in silica gel tlc
Yellow acid Rf' the corresponding colour band of value (GNA spot centers to parallax range/GA spot centers to parallax range) is uniformly divided into 4-6 sections,
And every section of sampling silica gel tlc method detects the purity of wherein GNA；
(4) merge above sterling containing GNA, the silica gel H compared with sterling respectively, 95% ethyl alcohol is cleaned molten after GNA in each section of silica gel
HPLC method detects purity again after the concentration of liquid decompression rotary evaporator, weighing, obtains GNA；
(5) merge the reception fraction containing pure GA in (2) test tube and depressurize rotary evaporated to dryness to get GA.
Discrete spectrum band is narrow and neat in the present invention, separating degree is big, can get compared with Gao Zhuxiao, with conventional wet silica gel column chromatography
Partition method, which is compared, that easy to operate, time-consuming is short, solvent-oil ratio is small, separative efficiency is high.
The spots such as GNA spot and GA R is found out with silica gel tlc method before mesolow gradient silica gel dry column chromatographyfDifference is larger, at
The good solvent of point property, and the eluant, eluent being directly used as in silica gel dry column chromatography.Silica gel H chromatographic column, mesolow are successively loaded again
The separation of gradient dry column chromatography, the elution silica gel H section of the sterling containing GNA, decompression concentrated solution again high performance liquid chromatography (with GNA
Standard control) it measures GNA purity and calculates the rate of recovery.
Through detecting, the present invention is using mesolow silica gel dry column chromatography device cheap in laboratory, successively with dichloroethanes-
Ethyl alcohol-triethylamine v/v 30:0.15:4 (eluant, eluent 1) and 20:0.6:3 (eluant, eluent 2) system are consolidated for eluant, eluent, with silica gel H
Determine phase, Jing Yici column chromatography can isolate the GNA sample of purity 93%, the rate of recovery inexpensive, easyly, fairly large
78%.
Embodiment 2
The method that EDCI.HCl, DMAP joint catalysis gambogicacid and arylamine reaction prepare N- aryl gamboge amide, including step
It is rapid as follows:
(1) GA (initial concentration of 4.9mmol/L), 1- ethyl -3- (N, N- dimethylamino) propyl carbodiimide hydrochloride
(initial concentration of EDCI.HCl, 1.1equiv., 5.5mmol/L), 4- (N, N- dimethyl) pyridine (DMAP, 0.2equiv.,
The initial concentration of 0.9mmol/L) and after 15min is stirred at room temperature in methylene chloride (DCM), the chloro- 3- 5-trifluoromethylaniline of 4- is added
The dichloromethane solution of (initial concentration of 1.1equiv., 4.8mmol/L).Room temperature continues to stir 11h, until silica gel tlc (expansion
Agent: dichloromethane-ethanol-triethylamine v/v 80:0.1:3) method detection reaction solution in gambogicacid spot completely disappear；
(2) plus after water quenching reaction organic layer is separated, it is dry that organic phase anhydrous magnesium sulfate is separated after washing (4mL × 2).Filter
Except desiccant filtrate decompression concentration after, crude product through silica gel H make stationary phase mesolow dry column chromatography can low cost, easily
Prepare N- (the chloro- 3- trifluoromethyl of 4-) phenyl gamboge amide sample of high-purity, yield 74~76%.
Embodiment 3
The method that EDCI.HCl, DMAP joint catalysis gambogicacid and arylamine reaction prepare N- aryl gamboge amide, including step
It is rapid as follows:
(1) GA (initial concentration of 4.9mmol/L), 1- ethyl -3- (N, N- dimethylamino) propyl carbodiimide hydrochloride
(initial concentration of EDCI.HCl, 1.1equiv., 5.5mmol/L), 4- (N, N- dimethyl) pyridine (DMAP, 0.2equiv.,
The initial concentration of 0.9mmol/L) and after 15min is stirred at room temperature in methylene chloride (DCM), 3- (6- chloropyridine base) amine is added
The dichloromethane solution of (initial concentration of 1.1equiv., 4.8mmol/L).Room temperature continues to stir 12h, until silica gel tlc (expansion
Agent: dichloromethane-ethanol-triethylamine v/v 80:0.1:3) method detection reaction solution in GA spot completely disappear；
(2) plus after water quenching reaction organic layer is separated, it is dry that organic phase anhydrous magnesium sulfate is separated after washing (4mL × 2).Filter
Except desiccant filtrate decompression concentration after, crude product through silica gel H make stationary phase mesolow dry column chromatography can low cost, easily
Prepare N- [3- (6- chloropyridine base)] gamboge amide sample of high-purity, yield 70~72%.
Comparative example 1
GA and GNA is separated using the common silica gel dry column chromatography of granularity 200-300 mesh (45-75 μm).
The absorption property of column chromatography silica gel depends on microcellular structure and bore area formed in its production process.Silica gel
The surface texture of inside particles hole is different from the skeletal internal structure of formation, constitution water contained by the silicon atom and colloid on surface
Form silanol group.The disequilibrium of this structure makes the surface of silica gel generate the free field of force, i.e., to hydrone or other polarity point
Son has adsorption capacity.Therefore silica gel has selectivity to the absorption of the mixture of different component.When the stronger component of molecular polarity
The adsorption capacity generated when passing through Silica Surface with silica gel is also relatively strong, and the component is longer in the retention time of Silica Surface；Opposite point
The weaker component of sub- polarity, retention time are shorter.Therefore the mixture of different component passes through the difference during silica gel because of retention time
Do not separated (Zhou Suhong, Zou Tao, old chromatographic silica gel absorption property of entangling inquire into the coloured mining and metallurgy of, 2006,22 (supplementary issues):
146-147)。
As the result is shown: 200-300 mesh silica gel, the more eluant, eluents of 2 times of weight, Cai Nengda are used under the conditions of same operation
Similar GNA separation purity (82%) when making stationary phase to silica gel H, and the rate of recovery is down to 56%.
Comparative example 2
The adverse current column chromatography method separation for adding sample to be separated, eluant, eluent to move from the bottom up using silica gel H bottom end.
As the result is shown: using the silica gel H of 1.5 times of weight in adverse current column chromatography, can be only achieved the dry column layer of mesolow gradient
Approximate GNA separation purity (87%) when analysis, and the rate of recovery is reduced to 64%.
Comparative example 3
The method that DCC, DMAP joint catalysis gambogicacid and arylamine reaction prepare N- aryl gamboge amide, including step is such as
Under:
(1) GA (initial concentration of 4.9mmol/L), N, N '-dicyclohexylcarbodiimide (DCC.HCl, 1.1equiv), 4-
(N, N- dimethyl) pyridine (initial concentration of DMAP, 0.2equiv., 0.9mmol/L), methylene chloride (DCM) are stirred at room temperature
After 18min, the dichloromethane solution of the chloro- 3- 5-trifluoromethylaniline of 4- (initial concentration of 1.1equiv., 4.8mmol/L) is added.
Room temperature continues to stir 28h, and silica gel tlc method (solvent: dichloromethane-ethanol-triethylamine v/v 80:0.1:3) detects reaction solution
Middle gambogicacid spot can not completely disappear.
(2) plus after water quenching reaction organic layer is separated, it is dry that organic phase anhydrous magnesium sulfate is separated after washing (4mL × 2).Filter
After the concentration of desiccant filtrate decompression, the column chromatography that crude product makees stationary phase through silica gel H obtains N- (the chloro- 3- trifluoromethyl of 4-) phenyl
Gamboge amide sample, yield 50~52%.
Due to easily deliquescing when DCC is weighed, not as good as EDCI.HCl, solubility is big in methylene chloride, DCC, DMAP joint catalysis
The amidated rate of GA virtue is catalyzed the rate of the reaction significantly lower than EDCI.HCl, DMAP method joint in embodiment 2.Additionally, due to
Solubility of by-product N, N '-dicyclohexylurea (DCU) of DCC catalysis reaction in methylene chloride, water, diluted acid is low, it is necessary to pass through column
Chromatographic runs could remove, and excessive EDCI.HCl and its side reaction product N- ethyl-N '-dimethylamino-propyl urea hydrochloride
Then easy-to-use dilute hydrochloric acid washing method removes.
In a word the catalysis GA virtue amidation of DCC, DMAP method joint not as good as EDCI.HCl, DMAP method reaction speed fast, high income,
And post-processing is also more cumbersome.
It although an embodiment of the present invention has been shown and described, for the ordinary skill in the art, can be with
Understand without departing from the principles and spirit of the present invention can to these examples carry out it is a variety of variation, modification, replacement and
Modification, the scope of the present invention is defined by the appended.
Claims (10)
1. the method that mesolow gradient silica gel dry column chromatography separates Neo-garcinolic acid, gambogicacid, comprises the following steps that
After the mother liquor acidification of gambogicacid pyridiniujm is precipitated after the medicinal extract of goa powder acetone extract successively pyridine, water temperature heat treatment
Acetic acid ethyl acetate extract be sample to be separated, stationary phase is made, with dichloro-ethane-ethanol-triethylamine triplet with silica gel H respectively
System is eluant, eluent, is separated using the flow direction of mesolow gradient silica gel H dry column chromatography eluant, eluent from top to bottom, makes new gamboge
Acid is separated into different bands of a spectrum on silica gel H column from ingredients such as gambogicacid B, impurity, and gambogicacid is then by whole from silica gel H column
Gambogicacid eluent is eluted and collects, ethanol washing silica gel H is simultaneously concentrated to get Neo-garcinolic acid；The concentration of gambogicacid eluent, i.e., separately
Obtain gambogicacid.
2. the method for mesolow gradient silica gel dry column chromatography separation Neo-garcinolic acid according to claim 1, gambogicacid, special
Sign is, eluant, eluent is divided into eluant, eluent 1 and eluant, eluent 2, dichloro-ethane-ethanol-triethylamine volume ratio is in eluant, eluent 1
30:0.15:4, dichloro-ethane-ethanol-triethylamine volume ratio is 20:0.6:3 in eluant, eluent 2；First with eluant, eluent 1 elution until
1 forward position of eluant, eluent ran silica gel H bottom and received the eluant, eluent 1 containing GA, the elution of eluant, eluent 2 was reused, until silica gel tlc
Method detection receives GA in fraction and is all eluted in gambogicacid eluent.
3. the method for mesolow silica gel dry column chromatography separation Neo-garcinolic acid according to claim 1, gambogicacid, feature exist
In silica gel H and example weight to be separated ratio are 60-90:1.
4. the method for mesolow silica gel dry column chromatography separation Neo-garcinolic acid according to claim 1, gambogicacid, feature exist
In silica gel H height and chromatography column internal diameter ratio are greater than 8:1.
5. the method for mesolow silica gel dry column chromatography separation Neo-garcinolic acid according to claim 1, gambogicacid, feature exist
In eluant, eluent is carried out by 3-4cm height/min or 1 drop/s speed.
6. the method for mesolow silica gel dry column chromatography separation Neo-garcinolic acid according to claim 1, gambogicacid, feature exist
In partial size < 38 μm of silica gel H.
7. the method for preparing N- aryl gamboge amide with the GA that method described in claim 1 is isolated, comprising the following steps:
(1) by GA, 1- ethyl -3- (N, N- dimethylamino) propyl carbodiimide hydrochloride (EDCI.HCl), 4- (N, N- diformazan ammonia
Base) pyridine (DMAP) and methylene chloride (DCM) mixing, be stirred at room temperature the dichloromethane solution that arylamine is added after 15min, room temperature after
Continuous stirring 6-12h GA spot into silica gel tlc method detection reaction solution completely disappears；
(2) plus after water quenching reaction organic layer is separated, the drying of organic phase anhydrous magnesium sulfate is separated after washing, filtrate decompression is through rotating
After evaporator concentration, crude product makees the mesolow gradient dry column chromatography of stationary phase through silica gel H to get the N- aryl gamboge of high-purity
Amide sample.
8. the method according to claim 7 for preparing N- aryl gamboge amide, which is characterized in that GA, 1- ethyl -3- (N,
N- dimethylamino) molar ratio of propyl carbodiimide hydrochloride (EDCI.HCl) and 4- (N, N- dimethylamino) pyridine (DMAP) is
1:(1.05-1.15): (0.2-0.4).
9. the method according to claim 7 for preparing N- aryl gamboge amide, which is characterized in that the molar ratio of GA and arylamine
For 1:(1.1-1.2).
10. the method according to claim 7 for preparing N- aryl gamboge amide, which is characterized in that the arylamine is 4-
Chloro- 3- 5-trifluoromethylaniline or 3- (6- chloropyridine base) amine.
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| TAO CHEN，等: "Synthesis and antiangiogenic activity of novel gambogic acid derivatives", 《MOLECULES》 * |
| 符雄: "有机天然产物的分离方法—快速柱层析", 《中山大学研究生学刊自然科学版》 * |
| 胡钟: "新藤黄酸衍生物的设计、合成及杭肿瘤活性初步研究", 《中国优秀硕士学位论文全文数据库 医药卫生科技辑》 * |
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