CN109485569A - A kind of acetoxyl group isopentene group substituted biphenyl class compound and its preparation method and application - Google Patents
A kind of acetoxyl group isopentene group substituted biphenyl class compound and its preparation method and application Download PDFInfo
- Publication number
- CN109485569A CN109485569A CN201810884454.8A CN201810884454A CN109485569A CN 109485569 A CN109485569 A CN 109485569A CN 201810884454 A CN201810884454 A CN 201810884454A CN 109485569 A CN109485569 A CN 109485569A
- Authority
- CN
- China
- Prior art keywords
- substituted biphenyl
- class compound
- group substituted
- medicinal extract
- acetoxyl group
- 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
- -1 acetoxyl group Chemical group 0.000 title claims abstract description 89
- YHQXBTXEYZIYOV-UHFFFAOYSA-N 3-methylbut-1-ene Chemical group CC(C)C=C YHQXBTXEYZIYOV-UHFFFAOYSA-N 0.000 title claims abstract description 40
- 238000002360 preparation method Methods 0.000 title claims abstract description 14
- 239000000284 extract Substances 0.000 claims abstract description 57
- 239000003960 organic solvent Substances 0.000 claims abstract description 46
- 241000546193 Clusiaceae Species 0.000 claims abstract description 16
- 235000013399 edible fruits Nutrition 0.000 claims abstract description 15
- 230000002953 anti-rotaviral effect Effects 0.000 claims abstract description 14
- 238000000638 solvent extraction Methods 0.000 claims abstract description 12
- 238000004128 high performance liquid chromatography Methods 0.000 claims abstract description 11
- 238000000605 extraction Methods 0.000 claims abstract description 10
- 238000010898 silica gel chromatography Methods 0.000 claims abstract description 10
- 239000003814 drug Substances 0.000 claims abstract description 9
- 239000002994 raw material Substances 0.000 claims abstract description 7
- OKKJLVBELUTLKV-UHFFFAOYSA-N Methanol Chemical compound OC OKKJLVBELUTLKV-UHFFFAOYSA-N 0.000 claims description 96
- 239000012071 phase Substances 0.000 claims description 38
- 239000003480 eluent Substances 0.000 claims description 35
- 239000000243 solution Substances 0.000 claims description 22
- VYPSYNLAJGMNEJ-UHFFFAOYSA-N Silicium dioxide Chemical compound O=[Si]=O VYPSYNLAJGMNEJ-UHFFFAOYSA-N 0.000 claims description 19
- 239000000741 silica gel Substances 0.000 claims description 19
- 229910002027 silica gel Inorganic materials 0.000 claims description 19
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 16
- XEKOWRVHYACXOJ-UHFFFAOYSA-N Ethyl acetate Chemical group CCOC(C)=O XEKOWRVHYACXOJ-UHFFFAOYSA-N 0.000 claims description 15
- 239000007864 aqueous solution Substances 0.000 claims description 15
- 238000010828 elution Methods 0.000 claims description 15
- CSCPPACGZOOCGX-UHFFFAOYSA-N Acetone Chemical compound CC(C)=O CSCPPACGZOOCGX-UHFFFAOYSA-N 0.000 claims description 12
- HEDRZPFGACZZDS-UHFFFAOYSA-N Chloroform Chemical group ClC(Cl)Cl HEDRZPFGACZZDS-UHFFFAOYSA-N 0.000 claims description 10
- 239000003208 petroleum Chemical group 0.000 claims description 10
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 claims description 9
- 238000001514 detection method Methods 0.000 claims description 9
- 238000001914 filtration Methods 0.000 claims description 8
- 239000000463 material Substances 0.000 claims description 8
- 239000013049 sediment Substances 0.000 claims description 8
- 238000000926 separation method Methods 0.000 claims description 8
- RTZKZFJDLAIYFH-UHFFFAOYSA-N Diethyl ether Chemical group CCOCC RTZKZFJDLAIYFH-UHFFFAOYSA-N 0.000 claims description 7
- 230000005526 G1 to G0 transition Effects 0.000 claims description 7
- UHOVQNZJYSORNB-UHFFFAOYSA-N Benzene Chemical compound C1=CC=CC=C1 UHOVQNZJYSORNB-UHFFFAOYSA-N 0.000 claims description 6
- 238000004440 column chromatography Methods 0.000 claims description 6
- 238000002137 ultrasound extraction Methods 0.000 claims description 5
- WORJEOGGNQDSOE-UHFFFAOYSA-N chloroform;methanol Chemical compound OC.ClC(Cl)Cl WORJEOGGNQDSOE-UHFFFAOYSA-N 0.000 claims description 4
- 235000019441 ethanol Nutrition 0.000 claims description 4
- SRCZQMGIVIYBBJ-UHFFFAOYSA-N ethoxyethane;ethyl acetate Chemical compound CCOCC.CCOC(C)=O SRCZQMGIVIYBBJ-UHFFFAOYSA-N 0.000 claims description 4
- HWJHWSBFPPPIPD-UHFFFAOYSA-N ethoxyethane;propan-2-one Chemical compound CC(C)=O.CCOCC HWJHWSBFPPPIPD-UHFFFAOYSA-N 0.000 claims description 4
- 239000007791 liquid phase Substances 0.000 claims description 4
- 238000004458 analytical method Methods 0.000 claims description 3
- 238000013375 chromatographic separation Methods 0.000 claims description 3
- 238000011097 chromatography purification Methods 0.000 claims description 3
- 235000010290 biphenyl Nutrition 0.000 claims description 2
- OAIVIYSBZFEOIU-UHFFFAOYSA-N chloroform;propan-2-one Chemical compound CC(C)=O.ClC(Cl)Cl OAIVIYSBZFEOIU-UHFFFAOYSA-N 0.000 claims description 2
- ZUOUZKKEUPVFJK-UHFFFAOYSA-N phenylbenzene Natural products C1=CC=CC=C1C1=CC=CC=C1 ZUOUZKKEUPVFJK-UHFFFAOYSA-N 0.000 claims description 2
- 239000004305 biphenyl Substances 0.000 claims 1
- 150000001875 compounds Chemical class 0.000 abstract description 43
- 230000000694 effects Effects 0.000 abstract description 10
- 238000002474 experimental method Methods 0.000 abstract description 6
- IWUCXVSUMQZMFG-AFCXAGJDSA-N Ribavirin Chemical compound N1=C(C(=O)N)N=CN1[C@H]1[C@H](O)[C@H](O)[C@@H](CO)O1 IWUCXVSUMQZMFG-AFCXAGJDSA-N 0.000 abstract description 4
- HZCAHMRRMINHDJ-DBRKOABJSA-N ribavirin Natural products O[C@@H]1[C@H](O)[C@@H](CO)O[C@H]1N1N=CN=C1 HZCAHMRRMINHDJ-DBRKOABJSA-N 0.000 abstract description 4
- 229960000329 ribavirin Drugs 0.000 abstract description 4
- 238000000034 method Methods 0.000 description 16
- 238000005481 NMR spectroscopy Methods 0.000 description 11
- OKKJLVBELUTLKV-MZCSYVLQSA-N Deuterated methanol Chemical compound [2H]OC([2H])([2H])[2H] OKKJLVBELUTLKV-MZCSYVLQSA-N 0.000 description 10
- IAZDPXIOMUYVGZ-UHFFFAOYSA-N Dimethylsulphoxide Chemical compound CS(C)=O IAZDPXIOMUYVGZ-UHFFFAOYSA-N 0.000 description 10
- 238000001644 13C nuclear magnetic resonance spectroscopy Methods 0.000 description 6
- OKTJSMMVPCPJKN-UHFFFAOYSA-N Carbon Chemical compound [C] OKTJSMMVPCPJKN-UHFFFAOYSA-N 0.000 description 6
- 229910052799 carbon Inorganic materials 0.000 description 6
- 235000015110 jellies Nutrition 0.000 description 6
- 239000008274 jelly Substances 0.000 description 6
- 125000000956 methoxy group Chemical group [H]C([H])([H])O* 0.000 description 6
- 241000593508 Garcinia Species 0.000 description 5
- 235000000885 Garcinia xanthochymus Nutrition 0.000 description 5
- 238000003919 heteronuclear multiple bond coherence Methods 0.000 description 5
- 239000000843 powder Substances 0.000 description 5
- 231100001274 therapeutic index Toxicity 0.000 description 5
- 241000196324 Embryophyta Species 0.000 description 4
- 238000004090 dissolution Methods 0.000 description 4
- 238000002114 high-resolution electrospray ionisation mass spectrometry Methods 0.000 description 4
- 229910052739 hydrogen Inorganic materials 0.000 description 4
- 125000002255 pentenyl group Chemical group C(=CCCC)* 0.000 description 4
- IOLCXVTUBQKXJR-UHFFFAOYSA-M potassium bromide Chemical compound [K+].[Br-] IOLCXVTUBQKXJR-UHFFFAOYSA-M 0.000 description 4
- 238000005160 1H NMR spectroscopy Methods 0.000 description 3
- HIXDQWDOVZUNNA-UHFFFAOYSA-N 2-(3,4-dimethoxyphenyl)-5-hydroxy-7-methoxychromen-4-one Chemical group C=1C(OC)=CC(O)=C(C(C=2)=O)C=1OC=2C1=CC=C(OC)C(OC)=C1 HIXDQWDOVZUNNA-UHFFFAOYSA-N 0.000 description 3
- 241000700605 Viruses Species 0.000 description 3
- 230000000259 anti-tumor effect Effects 0.000 description 3
- 239000004615 ingredient Substances 0.000 description 3
- 125000002496 methyl group Chemical group [H]C([H])([H])* 0.000 description 3
- 238000001228 spectrum Methods 0.000 description 3
- 230000004083 survival effect Effects 0.000 description 3
- 230000003612 virological effect Effects 0.000 description 3
- 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 description 2
- UFHFLCQGNIYNRP-UHFFFAOYSA-N Hydrogen Chemical compound [H][H] UFHFLCQGNIYNRP-UHFFFAOYSA-N 0.000 description 2
- 238000004566 IR spectroscopy Methods 0.000 description 2
- 241000702670 Rotavirus Species 0.000 description 2
- 238000010521 absorption reaction Methods 0.000 description 2
- 150000001555 benzenes Chemical group 0.000 description 2
- 150000008366 benzophenones Chemical class 0.000 description 2
- 238000006243 chemical reaction Methods 0.000 description 2
- 239000000470 constituent Substances 0.000 description 2
- WBKFWQBXFREOFH-UHFFFAOYSA-N dichloromethane;ethyl acetate Chemical compound ClCCl.CCOC(C)=O WBKFWQBXFREOFH-UHFFFAOYSA-N 0.000 description 2
- 238000001035 drying Methods 0.000 description 2
- 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 description 2
- 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 description 2
- 239000001257 hydrogen Substances 0.000 description 2
- 125000002887 hydroxy group Chemical group [H]O* 0.000 description 2
- 230000002401 inhibitory effect Effects 0.000 description 2
- 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 description 2
- 239000010410 layer Substances 0.000 description 2
- 239000007788 liquid Substances 0.000 description 2
- 125000001434 methanylylidene group Chemical group [H]C#[*] 0.000 description 2
- 125000000325 methylidene group Chemical group [H]C([H])=* 0.000 description 2
- 239000002356 single layer Substances 0.000 description 2
- 239000002904 solvent Substances 0.000 description 2
- 239000000126 substance Substances 0.000 description 2
- 230000001629 suppression Effects 0.000 description 2
- 238000012360 testing method Methods 0.000 description 2
- 238000002211 ultraviolet spectrum Methods 0.000 description 2
- MDBKCNBPGJPWMA-UHFFFAOYSA-N xanthen-9-one Chemical class C1=CC=C2C(=O)C3=CC=CC=C3OC2=C1.C1=CC=C2C(=O)C3=CC=CC=C3OC2=C1 MDBKCNBPGJPWMA-UHFFFAOYSA-N 0.000 description 2
- JNELGWHKGNBSMD-UHFFFAOYSA-N xanthone Chemical compound C1=CC=C2C(=O)C3=CC=CC=C3OC2=C1 JNELGWHKGNBSMD-UHFFFAOYSA-N 0.000 description 2
- OAVCWZUKQIEFGG-UHFFFAOYSA-O 2-(5-methyl-2H-tetrazol-1-ium-1-yl)-1,3-thiazole Chemical compound CC1=NN=N[NH+]1C1=NC=CS1 OAVCWZUKQIEFGG-UHFFFAOYSA-O 0.000 description 1
- 238000007445 Chromatographic isolation Methods 0.000 description 1
- 208000036142 Viral infection Diseases 0.000 description 1
- 238000002835 absorbance Methods 0.000 description 1
- 125000000218 acetic acid group Chemical group C(C)(=O)* 0.000 description 1
- 238000010171 animal model Methods 0.000 description 1
- 230000000840 anti-viral effect Effects 0.000 description 1
- 239000012965 benzophenone Substances 0.000 description 1
- 229940093797 bioflavonoids Drugs 0.000 description 1
- 230000004071 biological effect Effects 0.000 description 1
- 150000004074 biphenyls Chemical class 0.000 description 1
- 238000001460 carbon-13 nuclear magnetic resonance spectrum Methods 0.000 description 1
- 230000030833 cell death Effects 0.000 description 1
- 239000013553 cell monolayer Substances 0.000 description 1
- 239000006285 cell suspension Substances 0.000 description 1
- 238000004587 chromatography analysis Methods 0.000 description 1
- 230000001186 cumulative effect Effects 0.000 description 1
- 231100000433 cytotoxic Toxicity 0.000 description 1
- 230000001472 cytotoxic effect Effects 0.000 description 1
- 231100000135 cytotoxicity Toxicity 0.000 description 1
- 230000003013 cytotoxicity Effects 0.000 description 1
- YCJBWNIROIXYPD-UHFFFAOYSA-N depsidone Chemical compound O=C1OC2=CC=CC=C2OC2=CC=CC=C12 YCJBWNIROIXYPD-UHFFFAOYSA-N 0.000 description 1
- 201000010099 disease Diseases 0.000 description 1
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 1
- 229940079593 drug Drugs 0.000 description 1
- 238000011156 evaluation Methods 0.000 description 1
- 238000013401 experimental design Methods 0.000 description 1
- 239000001963 growth medium Substances 0.000 description 1
- 238000000338 in vitro Methods 0.000 description 1
- 238000011534 incubation Methods 0.000 description 1
- 208000015181 infectious disease Diseases 0.000 description 1
- 230000005764 inhibitory process Effects 0.000 description 1
- 238000002347 injection Methods 0.000 description 1
- 239000007924 injection Substances 0.000 description 1
- 230000003834 intracellular effect Effects 0.000 description 1
- 150000002576 ketones Chemical class 0.000 description 1
- 239000002609 medium Substances 0.000 description 1
- 230000000116 mitigating effect Effects 0.000 description 1
- 239000000203 mixture Substances 0.000 description 1
- 238000012544 monitoring process Methods 0.000 description 1
- 239000012452 mother liquor Substances 0.000 description 1
- 229930014626 natural product Natural products 0.000 description 1
- 230000010355 oscillation Effects 0.000 description 1
- 125000001820 oxy group Chemical group [*:1]O[*:2] 0.000 description 1
- 230000000144 pharmacologic effect Effects 0.000 description 1
- 239000002574 poison Substances 0.000 description 1
- 238000012545 processing Methods 0.000 description 1
- 230000035755 proliferation Effects 0.000 description 1
- 238000011160 research Methods 0.000 description 1
- 230000003595 spectral effect Effects 0.000 description 1
- 238000012306 spectroscopic technique Methods 0.000 description 1
- 230000001954 sterilising effect Effects 0.000 description 1
- 238000004659 sterilization and disinfection Methods 0.000 description 1
- 239000004575 stone Substances 0.000 description 1
- 238000006467 substitution reaction Methods 0.000 description 1
- 239000006228 supernatant Substances 0.000 description 1
- 231100000331 toxic Toxicity 0.000 description 1
- 231100000816 toxic dose Toxicity 0.000 description 1
- 230000002588 toxic effect Effects 0.000 description 1
- 230000009466 transformation Effects 0.000 description 1
- 238000002604 ultrasonography Methods 0.000 description 1
- 238000000825 ultraviolet detection Methods 0.000 description 1
- 230000009385 viral infection Effects 0.000 description 1
- 238000005406 washing Methods 0.000 description 1
- 150000007964 xanthones Chemical class 0.000 description 1
Classifications
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07C—ACYCLIC OR CARBOCYCLIC COMPOUNDS
- C07C69/00—Esters of carboxylic acids; Esters of carbonic or haloformic acids
- C07C69/02—Esters of acyclic saturated monocarboxylic acids having the carboxyl group bound to an acyclic carbon atom or to hydrogen
- C07C69/12—Acetic acid esters
- C07C69/21—Acetic acid esters of hydroxy compounds with more than three hydroxy groups
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P31/00—Antiinfectives, i.e. antibiotics, antiseptics, chemotherapeutics
- A61P31/12—Antivirals
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07C—ACYCLIC OR CARBOCYCLIC COMPOUNDS
- C07C67/00—Preparation of carboxylic acid esters
- C07C67/48—Separation; Purification; Stabilisation; Use of additives
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07C—ACYCLIC OR CARBOCYCLIC COMPOUNDS
- C07C67/00—Preparation of carboxylic acid esters
- C07C67/48—Separation; Purification; Stabilisation; Use of additives
- C07C67/56—Separation; Purification; Stabilisation; Use of additives by solid-liquid treatment; by chemisorption
Landscapes
- Organic Chemistry (AREA)
- Chemical & Material Sciences (AREA)
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- General Chemical & Material Sciences (AREA)
- Pharmacology & Pharmacy (AREA)
- Oncology (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Virology (AREA)
- Medicinal Chemistry (AREA)
- Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
- Communicable Diseases (AREA)
- Animal Behavior & Ethology (AREA)
- General Health & Medical Sciences (AREA)
- Public Health (AREA)
- Veterinary Medicine (AREA)
- Acyclic And Carbocyclic Compounds In Medicinal Compositions (AREA)
- Medicines Containing Plant Substances (AREA)
Abstract
The invention discloses a kind of acetoxyl group isopentene group substituted biphenyl class compound and its preparation method and application, the acetoxyl group isopentene group substituted biphenyl class compound molecule formula is C20H22O6, there is following structural formula:The preparation method is to dry Guttiferae arbor branch, leaf or fruit as raw material, through medicinal extract extraction, organic solvent extraction, silica gel column chromatography, high pressure liquid chromatography separating step.The application is that acetoxyl group isopentene group substituted biphenyl class compound is preparing the application in anti-rotavirus medicaments.Through anti-rotavirus activity experiment, select Ribavirin as control, CC of the compound to rotavirus50And EC50Value is respectively 175.5 and 32μMol/L, with preferable anti-rotavirus activity.The simple activity of the compounds of this invention structure is good, can be used as the guiding compound of anti-rotavirus medicaments, there is good application prospect.
Description
Technical field
The invention belongs to effective ingredients in plant extractive technique fields, and in particular to a kind of acetoxyl group isopentene group substitution connection
Benzene-like compounds and its preparation method and application.
Background technique
Guttiferae (Garcinia L.) about 450 kinds of the plant whole world, Asia, Africa south and Polynesia western part are produced, I
State-owned 21 kinds, it is distributed in the southern provinces and regions such as Guangdong, Guangxi, Yunnan.Garcinia maingayii or natural xanthone (xanthone) class
One of main resource of ingredient, rich in the xanthone (xanthones) that isopentene group replaces, this kind of constituent structure is novel and diversified,
And there is extensive pharmacological activity, and it is especially most representative with gambogicacid (gambogic acid), it is antitumor with broad spectrum high-effect
Activity is one of the research hotspot of Antitumor Natural Products in recent years, and Chinese scholar is developing its injection, is antitumor one
Kind new medicine.In addition to xanthone (xanthone) class, biphenyl class (biphenyls), Benzophenone class (benzophenones), double Huangs
The compounds such as ketone (bioflavonoids) and depsidone compound (depsidone) are also the characteristic of graminaceous plant
Ingredient, it may have multiple biological activities.In order to more effectively utilize China's Garcinia maingayii resource, therefrom find before there is exploitation
The active constituent of scape, we select to carry out Garcinia maingayii the study of active components work compared with system.
Summary of the invention
The first object of the present invention is to provide a kind of acetoxyl group isopentene group substituted biphenyl class compound；Second purpose exists
In the preparation method of the offer acetoxyl group isopentene group substituted biphenyl class compound；Third is designed to provide the acetyl
The application of oxygroup isopentene group substituted biphenyl class compound.
The first object of the present invention is achieved in that the acetoxyl group isopentene group substituted biphenyl class compound is
To dry Guttiferae arbor branch, leaf or fruit as raw material, through medicinal extract extraction, organic solvent extraction, silica gel column chromatography, efficient liquid
Phase chromatographic isolation obtains, which is C20H22O6, the compound is with following structural formula:
The second object of the present invention is achieved in that the preparation of the acetoxyl group isopentene group substituted biphenyl class compound
Method is extracted through medicinal extract, organic solvent extraction, silica gel column layer using drying Guttiferae arbor branch, leaf or fruit as raw material
Analysis, high pressure liquid chromatography separation obtain, specifically:
A, medicinal extract extracts: by Guttiferae arbor branch, leaf or fruit coarse crushing to 20 ~ 40 mesh, with organic solvent ultrasonic extraction 2 ~ 4
Secondary, 30 ~ 60min, extracting solution merge every time；Extracting solution filtering when extracting solution to 1/4 ~ 1/2 volume is concentrated under reduced pressure, is stood,
Sediment is filtered out, medicinal extract a is condensed into；
B, organic solvent extracts: the water of 1 ~ 2 times of weight ratio amount being added in medicinal extract a, with the organic solvent extraction 3 ~ 5 isometric with water
It is secondary, merge organic solvent extraction phase, is concentrated under reduced pressure into medicinal extract b；
C, silica gel column chromatography: the organic solvent that medicinal extract b is measured with 1.5 ~ 3 times of weight ratio dissolves, and then weighs 0.8 ~ 1.2 with medicinal extract
Times 200 ~ 300 mesh silica gel mixed samples, then upper silica gel column chromatography, dress column silica gel is 200 ~ 300 mesh, dosage for medicinal extract b weight 6 ~
8 times of amounts；The mixed organic solvents gradient elution for being 1:0 ~ 0:1 with volume ratio is collected gradient eluent, concentration, is monitored through TLC,
Merge identical part；
D, reversed phase column chromatography: reversed phase column chromatography on the eluent that the organic solvent matched with 4:1 is afforded, reverse phase
Column is with reversed material C-18, C-8 or ODS dress column；Gradient is carried out with the methanol aqueous solution that volume content is 70 ~ 100% to wash
It is de-, it collects each section eluent and is concentrated, monitored through TLC, merge identical part；
E, high performance liquid chromatography separation: will be with the eluent that 70 ~ 100% methanol aqueous solution of volume content affords through efficient liquid
Phase chromatographic separation and purification is to get the acetoxyl group isopentene group substituted biphenyl class compound.
Acetoxyl group isopentene group substituted biphenyl class compound of the present invention is separated for the first time, passes through nuclear magnetic resonance
It is determined as acetoxyl group isopentene group substituted biphenyl class compound with other spectroscopic technique measuring methods, and characterizes its specific structure
Are as follows:
The compound is light orange jelly；Ultraviolet spectra (solvent is methanol),λ max(log ε): 570 (1.84), 206
(3.95) nm；Infrared spectroscopy (pressing potassium bromide troche) νmax 3428, 2935, 1713, 1614, 1492, 1455, 1434,
1371, 1314, 1267, 1228, 1070, 1024, 948, 837, 585 cm-1；HRESIMS shows the compounds of this invention
Quasi-molecular ion peakm/z358.1430 [M]+(calculated value 358.1416), in conjunction with13C and1H H NMR spectroscopy (Fig. 1 and Fig. 2, carbon
It is C that spectrum hydrogen modal data ownership, which is shown in Table and 1) provides its molecular formula,20H22O6。1H NMR (CD3OD, 400 MHz) and13C NMR
(CD3OD, 100 MHz) data, it is shown in Table 1.
HRESIMS shows that its quasi-molecular ion peak is quasi-molecular ion peakm/z 358.1430 [M]+(calculated value is
358.1416), in conjunction with13C H NMR spectroscopy determines that molecular formula is C20H22O6, degree of unsaturation 10.The compound1H- and13C-NMR
Attribution data shows that it contains 20 carbon, including 1 methoxyl group, 2 methyl, 2 methylene, 6 methines, 9
Quaternary carbon.According to nuclear magnetic resonance data [δ H 6.53 (1H, s, H-6), 7.10 (1H, d,J=8.1 Hz, H-8), 6.86
(1H, d, J=8.1 Hz, H-9, H-11), 7.10 (1H, d,J=8.1Hz, H-12), 3.31 (2H, d, J=6.6
Hz, H-1 '), 5.44 (1H, t,J=6.5 Hz, H-2 '), 4.42 (2H, s, H-4 '), 1.53 (3H, s, H-
5 '), 3.88 (3H, s, 3-OMe), 2.08 (3H, s, 4 '-OAc);δ C 129.8 (C-1), 124.1 (C-2),
145.1 (C-3), 138.3(C-4), 147.8 (C-5), 113.8 (C-6), 134.8 (C-7), 131.5 (C-8),
115.7 (C-9), 156.9 (C-10), 115.3 (C-11), 131.2 (C-12), 26.8 (C-1 '), 130.8 (C-
2 '), 134.5 (C-3 '), 71.2 (C-4 '), 13.9 (C-5 '), 60.8 (4-OMe), 172.9,20.8 (4 '-
OAc)].According toδ H3.88 (OMe) withδ C The HMBC correlation of 138.3 (C-4), methoxyl group are located at C-4.H-1'(δ H
And C-2(3.31)δ C 124.1), C-1(δ C 129.8) and C-3(δ C 145.1) and H-2'(δ H And C-2(5.44)δ C
124.1) HMBC correlation shows there is acetoxyl group isopentene group at C-2.According to C-3(δ C 145.1), C-5(δ C 147.8),
C-10(δ C 156.9) signal and molecular formula, three hydroxyls are located at C-3, C-5, C -10.1H and13C H NMR spectroscopy discloses
Five substituted benzene ring of 1,2,3,4,5- [δ H6.53(1H, s, H-6) signal:δ C 129.8(C-1), 124.1(C-2), 145.1(C-
3), 138.3(C-4), 147.8(C-5), 113.6(C-6)], [δ H7.10(2H, d,J=8.1 Hz, H-8, H-12), 6.86
(2H, d,J=8.0Hz, H-9, H-11).δ C134.8(C-7), 131.5(C-8, C-12), 115.7(C-9, C-11), 156.9
(C-10)] substitute mode of the compound, is further demonstrated.So far, the structure of the compound is determined.
The third object of the present invention is achieved in that the acetoxyl group isopentene group substituted biphenyl class compound exists
Prepare the application in anti-rotavirus medicaments.Through anti-rotavirus activity experiment, select Ribavirin as control, the chemical combination
CC of the object to rotavirus50And EC50Value is respectively 175.5 and 32μMol/L, with preferable anti-rotavirus activity.
The simple activity of the compounds of this invention structure is good, can be used as the guiding compound of anti-rotavirus medicaments, before having good application
Scape.
Detailed description of the invention
Fig. 1 be compound carbon-13 nmr spectra (13C NMR)；
Fig. 2 be compound nuclear magnetic resonance spectroscopy (1H NMR)；
Fig. 3 is the main HMBC(→ of compound) it is related.
Specific embodiment
Below with reference to embodiment and attached drawing, the present invention is further illustrated, but is not subject in any way to the present invention
Limitation, based on present invention teach that made any transformation or improvement, each fall within protection scope of the present invention.
Acetoxyl group isopentene group substituted biphenyl class compound of the present invention is to dry Guttiferae arbor branch, leaf
Or fruit is raw material, the chemical combination isolated through medicinal extract extraction, organic solvent extraction, silica gel column chromatography, high pressure liquid chromatography
Object molecular formula is C20H22O6, there is following structural formula:
The second object of the present invention is achieved in that the preparation of the acetoxyl group isopentene group substituted biphenyl class compound
Method is extracted through medicinal extract, organic solvent extraction, silica gel column layer using drying Guttiferae arbor branch, leaf or fruit as raw material
Analysis, high pressure liquid chromatography separation obtain, specifically:
A, medicinal extract extracts: by Guttiferae arbor branch, leaf or fruit coarse crushing to 20 ~ 40 mesh, with organic solvent ultrasonic extraction 2 ~ 4
Secondary, 30 ~ 60min, extracting solution merge every time；Extracting solution filtering when extracting solution to 1/4 ~ 1/2 volume is concentrated under reduced pressure, is stood,
Sediment is filtered out, medicinal extract a is condensed into；
B, organic solvent extracts: the water of 1 ~ 2 times of weight ratio amount being added in medicinal extract a, with the organic solvent extraction 3 ~ 5 isometric with water
It is secondary, merge organic solvent extraction phase, is concentrated under reduced pressure into medicinal extract b；
C, silica gel column chromatography: the organic solvent that medicinal extract b is measured with 1.5 ~ 3 times of weight ratio dissolves, and then weighs 0.8 ~ 1.2 with medicinal extract
Times 200 ~ 300 mesh silica gel mixed samples, then upper silica gel column chromatography, dress column silica gel is 200 ~ 300 mesh, dosage for medicinal extract b weight 6 ~
8 times of amounts；The mixed organic solvents gradient elution for being 1:0 ~ 0:1 with volume ratio is collected gradient eluent, concentration, is monitored through TLC,
Merge identical part；
D, reversed phase column chromatography: reversed phase column chromatography on the eluent that the organic solvent matched with 4:1 is afforded, reverse phase
Column is with reversed material C-18, C-8 or ODS dress column；Gradient is carried out with the methanol aqueous solution that volume content is 70 ~ 100% to wash
It is de-, it collects each section eluent and is concentrated, monitored through TLC, merge identical part；
E, high performance liquid chromatography separation: will be with the eluent that 70 ~ 100% methanol aqueous solution of volume content affords through efficient liquid
Phase chromatographic separation and purification is to get the acetoxyl group isopentene group substituted biphenyl class compound.
Further, acetone, ethyl alcohol or methanol that organic solvent described in step A is 70 ~ 100%.
Further, organic solvent described in step B is ethyl acetate, chloroform, ether, petroleum ether or benzene.
Further, mixed organic solvents described in step C are chloroform-acetone, chloroform-methanol, petroleum ether-acetone or stone
Oily ether-ethyl acetate etc..
Further, the volume proportion of mixed organic solvents described in step C is 1:0,4:1,2:1,1:1,0:1.
Further, the purifying of high performance liquid chromatography separation described in E step is the stream using 80 ~ 90% methanol as mobile phase
Speed 2 ~ 5ml/min, 9.4 × 250 mm, 5 μm of reverse phase preparative column are stationary phase, and UV detector Detection wavelength is 254 nm, often
Secondary 45 ~ 60 μ L of sample introduction collects the chromatographic peak of 15 ~ 20min, is evaporated after repeatedly adding up to get the acetoxyl group isopentene group
Substituted biphenyl class compound.
Acetoxyl group isopentene group substituted biphenyl class compound of the present invention is in preparing anti-rotavirus medicaments
Using.
Garcinia maingayii of the present invention is not limited by area and kind, and the present invention may be implemented.
Embodiment 1
Dry Guttiferae arbor branch, leaf and/or fruit 5.5kg are taken, coarse powder is broken to 20 mesh, with 70% acetone ultrasonic extraction
4 times, each 30min, extracting solution merges；Extracting solution filtering, is concentrated under reduced pressure into the 1/4 of volume；It stands, filters out sediment, it is dense
Shorten 260g medicinal extract a into；260g water is added in medicinal extract a, is extracted 5 times with the isometric ethyl acetate of water, merges extraction phase, subtract
Pressure is condensed into 154g medicinal extract b；With 200-300 mesh silica gel 1500g fill column, in medicinal extract b be added 200g methanol dissolution, then plus
Enter 100-200 mesh silica gel 160g and mix sample, mixes upper prop after sample；It is respectively 1:0,20:1,9:1,8:2,7:3,3:2,1 with volume ratio:
1, the chloroform-methanol mixed organic solvents gradient elution of 1:2,0:1 is collected gradient eluent, concentration, is monitored through TLC, merges phase
Same part, obtains 9 parts, and the eluent c of the chloroform-methanol mixed organic solvents of volume ratio 7:3 is 13g；Use reverse phase
Material C -18 fills column, and reversed-phase column on eluent c carries out gradient elution with the methanol aqueous solution that volume content is 20 ~ 100%, collects
Each section eluent is simultaneously concentrated, and monitors through TLC, merges identical part；It takes with the elution of 70 ~ 100% methanol aqueous solution of volume content
Obtained eluent, then using 85% methanol as mobile phase, flow velocity 3ml/min, 9.4 × 250 mm, 5 μm of Alltima C18
Reverse phase preparative column is stationary phase, and UV detector Detection wavelength is 254 nm, and each 50 μ L of sample introduction collects the chromatography of 12min
Peak is evaporated after repeatedly adding up to get the acetoxyl group isopentene group substituted biphenyl class compound.
Embodiment 2
Dry Guttiferae arbor branch, leaf and/or fruit 3.5kg are taken, coarse powder is broken to 20 mesh, with aqueous 10% EtOH Sonicate
It extracts 3 times, each 20min, extracting solution merges；Extracting solution filtering, is concentrated under reduced pressure into the 1/3 of volume；It stands, filters out sediment,
It is condensed into 360g medicinal extract a；The water of 360g is added in medicinal extract a, is extracted 3 times with the isometric ethyl acetate of water, merges extraction
Phase is taken, is concentrated under reduced pressure into 120g medicinal extract b；Column is filled with 200-300 mesh silica gel 1200g, the methanol that 240g is added in medicinal extract b is molten
Then solution is added 100-200 mesh silica gel 120g and mixes sample, mixes upper prop after sample；It is respectively 1:0,20:1,9:1,8:2,3 with volume ratio:
2, the chloroform of 1:1,1:2,0:1-acetone mixed organic solvents gradient elution is collected gradient eluent, concentration, is monitored through TLC, closes
And identical part；The chloroform of volume ratio 9:1-acetone mixed organic solvents eluent c is 46g；It is filled with reversed material C-18
Column, reversed-phase column on eluent c carry out gradient elution with the methanol aqueous solution that volume content is 20 ~ 100%, collect each section elution
Liquid is simultaneously concentrated, and monitors through TLC, merges identical part；It takes and is washed with what 70 ~ 100% methanol aqueous solution of volume content afforded
De- liquid, then using 80% methanol as mobile phase, flow velocity 3ml/min, flow velocity 2 ~ 5ml/min, 9.4 × 250 mm, 5 μm of Alltima
C18 reverse phase preparative column is stationary phase, and UV detector Detection wavelength is 254 nm, and each 50 μ L of sample introduction collects the color of 20min
Spectral peak is evaporated after repeatedly adding up to get the acetoxyl group isopentene group substituted biphenyl class compound.
Embodiment 3
Dry Guttiferae arbor branch, leaf and/or fruit 6.5kg are taken, coarse powder is broken to 30 mesh, with aqueous 20% methanol ultrasound
It extracts 3 times, each 20min, extracting solution merges；Extracting solution filtering, is concentrated under reduced pressure into the 1/2 of volume；It stands, filters out sediment,
It is condensed into 675g medicinal extract a；The water of 700g is added in medicinal extract a, is extracted 4 times with the isometric chloroform of water, merges extraction phase,
342g medicinal extract b is concentrated under reduced pressure into；Column is filled with 200-300 mesh silica gel 3400g, the methanol dissolution of 900g is added in medicinal extract b, so
100-200 mesh silica gel 360g is added afterwards and mixes sample, mixes upper prop after sample；It is respectively 1:0,20:1,9:1,8:2,7:3,3 with volume ratio:
2, the dichloromethane-ethyl acetate mixed organic solvents gradient elution of 1:1,1:2,0:1 collects gradient eluent, concentration, warp
TLC monitoring, merges identical part；The eluent c of the dichloromethane-ethyl acetate mixed organic solvents of volume ratio 9:1 is
45g；Column is filled with reversed material ODS, reversed-phase column on eluent c carries out ladder with the methanol aqueous solution that volume content is 20 ~ 100%
Degree elution is collected each section eluent and is concentrated, monitors through TLC, merge identical part；It takes with 80 ~ 100% first of volume content
The eluent that alcohol solution affords, then using 88% methanol as mobile phase, flow velocity 3ml/min, 9.4 × 250 mm, 5 μm
Alltima C18 reverse phase preparative column is stationary phase, and UV detector Detection wavelength is 254nm, each 50 μ L of sample introduction, is collected
The chromatographic peak of 7min is evaporated after repeatedly adding up to get the acetoxyl group isopentene group substituted biphenyl class compound.
Embodiment 4
Dry Guttiferae arbor branch, leaf and/or fruit 5.9kg are taken, coarse powder is broken to 40 mesh, it is extracted 3 times with 90% ethyl alcohol,
Extracting solution merges；Extracting solution filtering, is concentrated under reduced pressure into the 1/4 of volume；It stands, filters out sediment, be condensed into 760g medicinal extract a；?
The water of 800g is added in medicinal extract a, with petroleum ether extraction 4 times isometric with water, merges extraction phase, is concentrated under reduced pressure into 305g medicinal extract
b；Column is filled with 200-300 mesh silica gel 3000g, the methanol dissolution of 300g is added in medicinal extract b, 100-200 mesh silica gel is then added
300g mixes sample, mixes upper prop after sample；It is respectively the petroleum of 1:0,20:1,9:1,8:2,7:3,3:2,1:1,1:2,0:1 with volume ratio
Ether-acetone mixed organic solvents gradient elution collects gradient eluent, concentration, monitors through TLC, merge identical part；Volume
Petroleum ether-acetone mixed organic solvents eluent c than 8:2 is 43g；Column, reverse phase on eluent c are filled with reversed material C-8
Column carries out gradient elution with the methanol aqueous solution that volume content is 80 ~ 100%, collects each section eluent and be concentrated, supervise through TLC
It surveys, merges identical part；The eluent afforded with 70 ~ 100% methanol aqueous solution of volume content is taken, then with 90% methanol
For mobile phase, flow velocity 2ml/min, 9.4 × 250 mm, 5 μm of Alltima C18 reverse phase preparative column is stationary phase, ultraviolet detection
Device Detection wavelength is 254nm, collects the chromatographic peak of 8min, is evaporated after repeatedly adding up to get the acetoxyl group isopentene group
Substituted biphenyl class compound.
Embodiment 5
Dry 5.6 kg of Guttiferae arbor branch, leaf and/or fruit is taken, coarse powder is broken to 20 mesh, with 80% methanol ultrasonic extraction
3 times, each 30min, extracting solution merges；Extracting solution filtering, is concentrated under reduced pressure into the 1/2 of volume；It stands, filters out sediment, be concentrated
At 430g medicinal extract a；The water of 800g is added in medicinal extract a, with acetone extract 4 times isometric with water, merges extraction phase, depressurizes dense
Shorten 260g medicinal extract b into；Column is filled with 200-300 mesh silica gel 2200g, the ethyl acetate dissolution of 420g is added in medicinal extract b, then
100-200 mesh silica gel 400g is added and mixes sample, mixes upper prop after sample；With volume ratio be respectively 1:0,20:1,9:1,8:2,3:2,1:1,
The petroleum ether-ethyl acetate mixed organic solvents gradient elution of 1:2,0:1 are collected gradient eluent, concentration, are monitored through TLC, closes
And identical part；The eluent c of the petroleum ether-ethyl acetate mixed organic solvents of volume ratio 9:1 is 26g；Use reversed material
ODS fills column, and reversed-phase column on eluent c carries out gradient elution with the methanol aqueous solution that volume content is 20 ~ 100%, collects each portion
Divide eluent and be concentrated, is monitored through TLC, merge identical part；It takes and is afforded with 70 ~ 100% methanol aqueous solution of volume content
Eluent, then using 83% methanol as mobile phase, flow velocity 3ml/min, 9.4 × 250 mm, 5 μm of Alltima C18 reverse phase system
Standby column is stationary phase, and UV detector Detection wavelength is 254nm, collects the chromatographic peak of 17min, be evaporated after repeatedly cumulative to get
The acetoxyl group isopentene group substituted biphenyl class compound.
Embodiment 6
Compound prepared by Example 1 is light orange jelly；Measuring method are as follows: nuclear magnetic resonance is used, in conjunction with other wave spectrum skills
Art identifies structure.
(1) ultraviolet spectra (solvent is methanol),λ max(log ε): 570 (1.84), 206 (3.95) nm；
(2) infrared spectroscopy (pressing potassium bromide troche) νmax 3428, 2935, 1713, 1614, 1492, 1455, 1434,
1371, 1314, 1267, 1228, 1070, 1024, 948, 837, 585 cm-1；
(3) HRESIMS shows the compounds of this invention quasi-molecular ion peakm/z358.1430 [M]+(calculated value 358.1416),
In conjunction with13C and1(Fig. 1 and Fig. 2, it is C that carbon spectrum hydrogen modal data ownership is shown in Table and 1) provides its molecular formula to H H NMR spectroscopy20H22O6。1H NMR
(CD3OD, 400 MHz) and13C NMR(CD3OD, 100 MHz) data, it is shown in Table 1.
1 compound of table1H and13C NMR data (400/100 MHz, CD3OD)
HRESIMS shows that its quasi-molecular ion peak is quasi-molecular ion peakm/z 358.1430 [M]+(calculated value is
358.1416), in conjunction with13C H NMR spectroscopy determines that molecular formula is C20H22O6, degree of unsaturation 10.The compound1H- and13C-NMR
Attribution data shows that it contains 20 carbon, including 1 methoxyl group, 2 methyl, 2 methylene, 6 methines, 9
Quaternary carbon.According to nuclear magnetic resonance data [δ H 6.53 (1H, s, H-6), 7.10 (1H, d,J=8.1 Hz, H-8), 6.86
(1H, d, J=8.1 Hz, H-9, H-11), 7.10 (1H, d,J=8.1Hz, H-12), 3.31 (2H, d, J=6.6
Hz, H-1'), 5.44(1H, t, J=6.5 Hz, H-2'), 4.42 (2H, s, H-4'), 1.53 (3H, s, H-
5'), 3.88 (3H,s, 3-OMe), 2.08 (3H, s, 4'-OAc); δ C 129.8 (C-1), 124.1 (C-2),
145.1 (C-3), 138.3(C-4), 147.8 (C-5), 113.8 (C-6), 134.8 (C-7), 131.5 (C-8),
115.7 (C-9), 156.9 (C-10), 115.3 (C-11), 131.2 (C-12), 26.8 (C-1'), 130.8 (C-
2'), 134.5 (C-3'), 71.2 (C-4'), 13.9 (C-5'), 60.8 (4-OMe), 172.9, 20.8 (4'-
OAc)] according toδ H3.88 (OMe) withδ C The HMBC correlation of 138.3 (C-4), methoxyl group are located at C-4.H-1'(δ H
And C-2(3.31)δ C 124.1), C-1(δ C 129.8) and C-3(δ C 145.1) and H-2'(δ H And C-2(5.44)δ C
124.1) HMBC correlation shows there is acetoxyl group isopentene group at C-2.According to C-3(δ C 145.1), C-5(δ C 147.8),
C-10(δ C 156.9) signal and molecular formula, three hydroxyls are located at C-3, C-5, C -10.1H and13C H NMR spectroscopy discloses
Five substituted benzene ring of 1,2,3,4,5- [δ H6.53(1H, s, H-6) signal:δ C 129.8(C-1), 124.1(C-2), 145.1(C-
3), 138.3(C-4), 147.8(C-5), 113.6(C-6)], [δ H7.10(2H, d,J=8.1 Hz, H-8, H-12), 6.86
(2H, d,J=8.0Hz, H-9, H-11).δ C134.8(C-7), 131.5(C-8, C-12), 115.7(C-9, C-11), 156.9
(C-10)] substitute mode of the compound, is further demonstrated.So far, the structure of the compound is determined.
Embodiment 7
Compound prepared by Example 2 is light orange jelly；Structure determination is carried out by the method in embodiment 6, as a result
Are as follows: its structure is the same as embodiment 6, molecular formula C20H22O6.Confirm that compound prepared by embodiment 2 is that the acetoxyl group is different
Pentenyl substituted biphenyl class compound.
Embodiment 8
Compound prepared by Example 3 is light orange jelly；Structure determination is carried out by the method in embodiment 6, as a result
Are as follows: its structure is the same as embodiment 6, molecular formula C20H22O6.Confirm that compound prepared by embodiment 3 is that the acetoxyl group is different
Pentenyl substituted biphenyl class compound.
Embodiment 9
Compound prepared by Example 4 is light orange jelly；Structure determination is carried out by the method in embodiment 6, as a result
Are as follows: its structure is the same as embodiment 6, molecular formula C20H22O6.Confirm that compound prepared by embodiment 4 is that the acetoxyl group is different
Pentenyl substituted biphenyl class compound.
Embodiment 10
Compound prepared by Example 5 is light orange jelly；Structure determination is carried out by the method in embodiment 6, as a result
Are as follows: its structure is the same as embodiment 6, molecular formula C20H22O6.Confirm that compound prepared by embodiment 5 is that the acetoxyl group is different
Pentenyl substituted biphenyl class compound.
Embodiment 11
It is living to carry out anti-rotavirus for any acetoxyl group isopentene group substituted biphenyl class compound prepared by Example 1 ~ 5
Property detection test, test situation is as follows:
Cell strain: RhMK system (MA-104).
Experimental design: MA-104 cell and various concentration compound incubation 72 hours, the experiment of every plant of cell repeat 2
Secondary, the result tested with 3 times carries out data processing, using the inhibition journey of improvement MTT method evaluation compound on intracellular proliferation
Degree calculates inhibiting rate, calculates IC using Logit method according to inhibiting rate50, compare the Anti-viral activity in vitro of compound.
EC50 Refer to medium effective concentration, refers to and 50% experimental animal is caused to generate a certain specific reaction or certain reaction
Index is suppressed concentration when half.
CC50Refer to half cytotoxic concentration, refers to concentration needed for generating toxic effect to half cell.In this experiment
In, refer to drug concentration needed for causing 50% cell death.
(a) Compound Cytotoxicity measures
Compound is dissolved with dimethyl sulfoxide (dimethylsulfoxide, DMSO), in microwave sterilization l0min, uses MEM
The mother liquor for being made into lmg/ml is spare, and MEM solution is diluted to required concentration.96 porocyte culture plates add l x l05 /ml
The Mal04 cell suspension of concentration, 100uThe hole l/, 37 DEG C, 5%CO2 Incubator culture for 24 hours, on well-grown cell monolayer
Being separately added into concentration is respectively l mg/ml, 0.2 mg/ml, 40ug/ml、8 ug/ml、1.25 uThe compound of g/ml;100uThe hole l/, each concentration sets 3 multiple holes, while setting normal cell controls.37 DEG C are placed in, 5%CO2 Incubator continues culture for 24 hours
Afterwards, MTT method detects cell survival rate.
(b) compounds on viral infection mitigation acts on
With cell concentration for 104/ ml, every hole l00uL inoculating cell is cultivated 24 hours in 96 orifice plates, sees that cell is long
At single layer, simultaneously upgrowth situation is good, is respectively 100 with concentrationug/ml、75 ug/ml、50 ug/ml、25 ug/ml、l ug/
The compound of ml is after 37 DEG C of preparatory function cells of incubator l.5 h, PBS washing with the rotavirus of 100TCID50/ml
Every hole 100uIt is discarded after l absorption lh, adds MEM culture medium 100uThe hole l/ maintains, and sets 37C, 5%CO2Incubator, it is daily to see
Examine cytopathy situation.Viral suppression is detected with MTT method after 48h.
(c) compounds on viral treatment of infection acts on
With cell concentration for 104/ ml, every hole l00uL inoculating cell is cultivated 24 hours in 96 orifice plates, sees that cell is long
At single layer, simultaneously upgrowth situation is good, first the every hole 100 of rotavirus to 100TCID50/mluIt is abandoned after l absorption lh
It goes, is then added the compound of above-mentioned various concentration, 100uThe hole l/, ibid method culture and detection.It is right that each group experiment is all provided with virus
According to group (C group) and normal cell controls group (N group).
(d) cell survival rate measures
Using MTT method, 5mg/ml methyl thiazolyl tetrazolium (methyl is added in the cell of culture 48h
Thiazolyl tetrazolium, MTT) 20uL continues to cultivate 3-4 h, abandons supernatant, the every hole 100 DMSO is addeduL,
Oscillation make in the hole in crystallize be completely dissolved after immediately in 490luAbsorbance A value is measured under n wavelength.
Cell survival rate=medicine group is averaged A value/cell controls group A value x100%
Viral suppression=[experimental group be averaged one virus control group average A-value of A value]/[cell controls group is averaged one virus of A value
Control group is averaged A value] x 100%
Therapeutic index (TI)=half toxic concentration (CC50)/half-inhibitory concentration (IC50)
(e) experimental result
The results showed that selecting Ribavirin as control, the compound is to colyliform disease through anti-rotavirus activity experiment
The CC of poison50And EC50Value is respectively 175.5 and 32μMol/L is shown in Table 2, with preferable anti-rotavirus activity.
The anti-rotavirus activity of 2 compound of table
| No. | CC50 (µM) | EC50 (µM) | TI |
| The compound | 175.5+1.7 | 32+1.6 | 5.48 |
| Ribavirin | 263.2+1.9 | 13.3+0.7 | 19.8 |
aAll data are represented as average value ± SD(standard deviation); n = 3
TI: therapeutic index, CC50/EC50。
Claims (8)
1. a kind of acetoxyl group isopentene group substituted biphenyl class compound, it is characterised in that the acetoxyl group isopentene group takes
It is to dry Guttiferae arbor branch, leaf or fruit as raw material for biphenyl compound, extracted through medicinal extract extraction, organic solvent,
Silica gel column chromatography, high pressure liquid chromatography are isolated, which is C20H22O6, there is following structural formula:
。
2. a kind of preparation method of acetoxyl group isopentene group substituted biphenyl class compound according to claim 1, special
Sign is that dry Guttiferae arbor branch, leaf or fruit as raw material, is extracted through medicinal extract, organic solvent extraction, silica gel column layer
Analysis, high pressure liquid chromatography separation obtain, specifically:
A, medicinal extract extracts: by Guttiferae arbor branch, leaf or fruit coarse crushing to 20 ~ 40 mesh, with organic solvent ultrasonic extraction 2 ~ 4
Secondary, 30 ~ 60min, extracting solution merge every time；Extracting solution filtering when extracting solution to 1/4 ~ 1/2 volume is concentrated under reduced pressure, is stood,
Sediment is filtered out, medicinal extract a is condensed into；
B, organic solvent extracts: the water of 1 ~ 2 times of weight ratio amount being added in medicinal extract a, with the organic solvent extraction 3 ~ 5 isometric with water
It is secondary, merge organic solvent extraction phase, is concentrated under reduced pressure into medicinal extract b；
C, silica gel column chromatography: the organic solvent that medicinal extract b is measured with 1.5 ~ 3 times of weight ratio dissolves, and then weighs 0.8 ~ 1.2 with medicinal extract
Times 200 ~ 300 mesh silica gel mixed samples, then upper silica gel column chromatography, dress column silica gel is 200 ~ 300 mesh, dosage for medicinal extract b weight 6 ~
8 times of amounts；The mixed organic solvents gradient elution for being 1:0 ~ 0:1 with volume ratio is collected gradient eluent, concentration, is monitored through TLC,
Merge identical part；
D, reversed phase column chromatography: reversed phase column chromatography on the eluent that the organic solvent matched with 4:1 is afforded, reverse phase
Column is with reversed material C-18, C-8 or ODS dress column；Gradient is carried out with the methanol aqueous solution that volume content is 70 ~ 100% to wash
It is de-, it collects each section eluent and is concentrated, monitored through TLC, merge identical part；
E, high performance liquid chromatography separation: will be with the eluent that 70 ~ 100% methanol aqueous solution of volume content affords through efficient liquid
Phase chromatographic separation and purification is to get the acetoxyl group isopentene group substituted biphenyl class compound.
3. the preparation method of acetoxyl group isopentene group substituted biphenyl class compound, feature according to claim 2
It is that organic solvent described in step A is 70 ~ 100% acetone, ethyl alcohol or methanol.
4. the preparation method of acetoxyl group isopentene group substituted biphenyl class compound, feature according to claim 2
It is that organic solvent described in step B is ethyl acetate, chloroform, ether, petroleum ether or benzene.
5. the preparation method of acetoxyl group isopentene group substituted biphenyl class compound, feature according to claim 2
It is that mixed organic solvents described in step C are chloroform-acetone, chloroform-methanol, petroleum ether-acetone or petroleum ether-ethyl acetate
Deng.
6. the preparation method of acetoxyl group isopentene group substituted biphenyl class compound, feature according to claim 2
The volume proportion for being mixed organic solvents described in step C is 1:0,4:1,2:1,1:1,0:1.
7. the preparation method of acetoxyl group isopentene group substituted biphenyl class compound, feature according to claim 2
It is that the purifying of high performance liquid chromatography separation described in E step is 2 ~ 5ml/min of flow velocity using 80 ~ 90% methanol as mobile phase,
9.4 × 250 mm, 5Reverse phase preparative column be stationary phase, UV detector Detection wavelength is 254 nm, each sample introduction 45 ~
60 μ L collect the chromatographic peak of 15 ~ 20min, are evaporated after repeatedly adding up to get the acetoxyl group isopentene group substituted biphenyl class
Compound.
8. a kind of acetoxyl group isopentene group substituted biphenyl class compound described in claim 1 is preparing anti-rotavirus medicaments
In application.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201810884454.8A CN109485569B (en) | 2018-08-06 | 2018-08-06 | Acetoxy isopentene group substituted biphenyl compound and preparation method and application thereof |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201810884454.8A CN109485569B (en) | 2018-08-06 | 2018-08-06 | Acetoxy isopentene group substituted biphenyl compound and preparation method and application thereof |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN109485569A true CN109485569A (en) | 2019-03-19 |
| CN109485569B CN109485569B (en) | 2021-05-18 |
Family
ID=65689155
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201810884454.8A Expired - Fee Related CN109485569B (en) | 2018-08-06 | 2018-08-06 | Acetoxy isopentene group substituted biphenyl compound and preparation method and application thereof |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN109485569B (en) |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN113880857A (en) * | 2021-11-11 | 2022-01-04 | 山东大学 | Polyisopentenyl substituted cage-shaped xanthone compound and preparation method and application thereof |
Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105175383A (en) * | 2015-08-12 | 2015-12-23 | 云南民族大学 | Biphenyl compound and preparation method and application thereof |
| CN106831365A (en) * | 2016-12-27 | 2017-06-13 | 云南民族大学 | A kind of hydroxymethoxy substituted biphenyl class compound and its preparation method and application |
| CN106928170A (en) * | 2016-12-27 | 2017-07-07 | 云南民族大学 | A kind of dihydrofuran biphenyl compound and its preparation method and application |
-
2018
- 2018-08-06 CN CN201810884454.8A patent/CN109485569B/en not_active Expired - Fee Related
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105175383A (en) * | 2015-08-12 | 2015-12-23 | 云南民族大学 | Biphenyl compound and preparation method and application thereof |
| CN106831365A (en) * | 2016-12-27 | 2017-06-13 | 云南民族大学 | A kind of hydroxymethoxy substituted biphenyl class compound and its preparation method and application |
| CN106928170A (en) * | 2016-12-27 | 2017-07-07 | 云南民族大学 | A kind of dihydrofuran biphenyl compound and its preparation method and application |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN113880857A (en) * | 2021-11-11 | 2022-01-04 | 山东大学 | Polyisopentenyl substituted cage-shaped xanthone compound and preparation method and application thereof |
Also Published As
| Publication number | Publication date |
|---|---|
| CN109485569B (en) | 2021-05-18 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN104860912B (en) | Dimer ketone compound and preparation method and application thereof | |
| Gadetskaya et al. | Sulfated phenolic compounds from Limonium caspium: Isolation, structural elucidation, and biological evaluation | |
| CN105884621B (en) | A kind of sesquiterpenoids and its preparation method and application | |
| CN105348192B (en) | Isoquinoline alkaloids bases compound of antiviral activity and preparation method thereof in a kind of wing pod Cassia tora | |
| CN105175383B (en) | A kind of biphenyl compound and its preparation method and application | |
| CN104761526B (en) | A kind of isoflavonoid with antiviral activity and its preparation method and application | |
| CN105440092B (en) | The fast preparation method of flavonoid glycoside in a kind of Extracted From Oil-tea-cake | |
| Xu et al. | Flavonoids from the capitula of Eriocaulon australe | |
| CN106831365A (en) | A kind of hydroxymethoxy substituted biphenyl class compound and its preparation method and application | |
| CN106588838A (en) | Hydroxydihydrobovolide in portulaca oleracea, and extraction and separation method of hydroxydihydrobovolide | |
| Moradi-Afrapoli et al. | Bisabololoxide derivatives from Artemisia persica, and determination of their absolute configurations by ECD | |
| CN106957322B (en) | A kind of isobenzofuran class compound and the preparation method and application thereof that can improve cigarette smoking throat comfort | |
| Yuan et al. | Alopecuroidines A− C, three matrine-derived alkaloids from the seeds of Sophora alopecuroides | |
| CN106928170A (en) | A kind of dihydrofuran biphenyl compound and its preparation method and application | |
| CN109485569A (en) | A kind of acetoxyl group isopentene group substituted biphenyl class compound and its preparation method and application | |
| CN106008422B (en) | A kind of benzo lactone compound, its preparation method and the application in anticancer drug is prepared | |
| CN107056743B (en) | A kind of depsidone compound and its preparation method and application | |
| CN106916160B (en) | A kind of isobenzofuran class compounds process for production thereof that can improve cigarette suction comfort in pueraria lobata | |
| CN103232427B (en) | Xanthone compound as well as preparation method and application thereof | |
| CN105884588B (en) | One kind drop sesquiterpenoids and preparation method and application | |
| CN109485570A (en) | A kind of isopentene group substituted biphenyl class compound and its preparation method and application | |
| Wang et al. | Sesquiterpene lactones from Artemisia verlotorum and their anti-inflammatory activities | |
| CN109232681A (en) | The separation of verbascoside and identification method in a kind of sweet osmanthus | |
| CN104829580A (en) | Isoflavone compound contained in tobacco and preparation method and application thereof | |
| CN106008219B (en) | A kind of sesquiterpenoids, its preparation method and the application in anti-rotavirus medicaments are prepared |
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 | ||
| CF01 | Termination of patent right due to non-payment of annual fee |
Granted publication date: 20210518 |