CN105968150A - Preparation method for 7-O-ethylmorroniside - Google Patents
Preparation method for 7-O-ethylmorroniside Download PDFInfo
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- CN105968150A CN105968150A CN201610309791.5A CN201610309791A CN105968150A CN 105968150 A CN105968150 A CN 105968150A CN 201610309791 A CN201610309791 A CN 201610309791A CN 105968150 A CN105968150 A CN 105968150A
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- morroniside
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- fructus corni
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- IRKFOLIBBQDADK-LEYKIWRXSA-N methyl (1S,3R,4aS,8S,8aS)-3-ethoxy-1-methyl-8-[(2S,3R,4S,5S,6R)-3,4,5-trihydroxy-6-(hydroxymethyl)oxan-2-yl]oxy-1,3,4,4a,8,8a-hexahydropyrano[3,4-c]pyran-5-carboxylate Chemical compound CCO[C@H]1C[C@H]2[C@@H]([C@H](C)O1)[C@H](O[C@@H]1O[C@H](CO)[C@@H](O)[C@H](O)[C@H]1O)OC=C2C(=O)OC IRKFOLIBBQDADK-LEYKIWRXSA-N 0.000 title claims abstract description 62
- 238000002360 preparation method Methods 0.000 title claims abstract description 27
- 238000010438 heat treatment Methods 0.000 claims abstract description 15
- 238000000746 purification Methods 0.000 claims abstract description 13
- YMWUJEATGCHHMB-UHFFFAOYSA-N Dichloromethane Chemical compound ClCCl YMWUJEATGCHHMB-UHFFFAOYSA-N 0.000 claims description 39
- YTZSBJLNMIQROD-SFBCHFHNSA-N Morroniside Chemical compound O([C@@H]1OC=C([C@H]2C[C@H](O)O[C@@H](C)[C@H]21)C(=O)OC)[C@@H]1O[C@H](CO)[C@@H](O)[C@H](O)[C@H]1O YTZSBJLNMIQROD-SFBCHFHNSA-N 0.000 claims description 29
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 claims description 20
- XEKOWRVHYACXOJ-UHFFFAOYSA-N Ethyl acetate Chemical compound CCOC(C)=O XEKOWRVHYACXOJ-UHFFFAOYSA-N 0.000 claims description 17
- 238000004090 dissolution Methods 0.000 claims description 17
- YTZSBJLNMIQROD-UHFFFAOYSA-N (4aS)-1c-beta-D-glucopyranosyloxy-6xi-hydroxy-8t-methyl-(4ar,8ac)-5,6,8,8a-tetrahydro-1H,4aH-pyrano[3,4-c]pyran-4-carboxylic acid methyl ester Natural products C12C(C)OC(O)CC2C(C(=O)OC)=COC1OC1OC(CO)C(O)C(O)C1O YTZSBJLNMIQROD-UHFFFAOYSA-N 0.000 claims description 15
- 238000002425 crystallisation Methods 0.000 claims description 14
- 230000008025 crystallization Effects 0.000 claims description 14
- ZUKLFFYDSALIQW-MSUKCBDUSA-N Iridoid glycoside Chemical compound [H][C@]12CC[C@H](C(O)=O)[C@@]1([H])[C@H](OC1O[C@H](CO)[C@@H](O)[C@H](O)[C@H]1O)OC=C2 ZUKLFFYDSALIQW-MSUKCBDUSA-N 0.000 claims description 13
- 229930182489 iridoid glycoside Natural products 0.000 claims description 13
- AMBQHHVBBHTQBF-UHFFFAOYSA-N Loganin Natural products C12C(C)C(O)CC2C(C(=O)OC)=COC1OC1OC(CO)C(O)C(O)C1O AMBQHHVBBHTQBF-UHFFFAOYSA-N 0.000 claims description 10
- AMBQHHVBBHTQBF-UOUCRYGSSA-N loganin Chemical compound O([C@@H]1OC=C([C@H]2C[C@H](O)[C@H](C)[C@H]21)C(=O)OC)[C@@H]1O[C@H](CO)[C@@H](O)[C@H](O)[C@H]1O AMBQHHVBBHTQBF-UOUCRYGSSA-N 0.000 claims description 10
- 238000001035 drying Methods 0.000 claims description 8
- 238000010992 reflux Methods 0.000 claims description 7
- VEXZGXHMUGYJMC-UHFFFAOYSA-N Hydrochloric acid Chemical compound Cl VEXZGXHMUGYJMC-UHFFFAOYSA-N 0.000 claims description 6
- NBIIXXVUZAFLBC-UHFFFAOYSA-N Phosphoric acid Chemical compound OP(O)(O)=O NBIIXXVUZAFLBC-UHFFFAOYSA-N 0.000 claims description 6
- 239000002994 raw material Substances 0.000 claims description 5
- QAOWNCQODCNURD-UHFFFAOYSA-N Sulfuric acid Chemical compound OS(O)(=O)=O QAOWNCQODCNURD-UHFFFAOYSA-N 0.000 claims description 4
- 238000000605 extraction Methods 0.000 claims description 4
- 229930182470 glycoside Natural products 0.000 claims description 4
- 150000002338 glycosides Chemical class 0.000 claims description 4
- 229910000147 aluminium phosphate Inorganic materials 0.000 claims description 3
- GRYLNZFGIOXLOG-UHFFFAOYSA-N Nitric acid Chemical compound O[N+]([O-])=O GRYLNZFGIOXLOG-UHFFFAOYSA-N 0.000 claims description 2
- 230000021736 acetylation Effects 0.000 claims description 2
- 238000006640 acetylation reaction Methods 0.000 claims description 2
- 239000002260 anti-inflammatory agent Substances 0.000 claims description 2
- 229940124599 anti-inflammatory drug Drugs 0.000 claims description 2
- 239000003153 chemical reaction reagent Substances 0.000 claims description 2
- 239000012141 concentrate Substances 0.000 claims description 2
- 229910017604 nitric acid Inorganic materials 0.000 claims description 2
- 239000001117 sulphuric acid Substances 0.000 claims description 2
- 235000011149 sulphuric acid Nutrition 0.000 claims description 2
- 230000003110 anti-inflammatory effect Effects 0.000 abstract description 15
- 238000002474 experimental method Methods 0.000 abstract description 12
- 238000006243 chemical reaction Methods 0.000 abstract description 11
- 238000000034 method Methods 0.000 abstract description 7
- 239000003814 drug Substances 0.000 abstract description 4
- 238000012216 screening Methods 0.000 abstract description 4
- 230000015572 biosynthetic process Effects 0.000 abstract description 3
- 238000013461 design Methods 0.000 abstract description 3
- 239000012535 impurity Substances 0.000 abstract description 3
- 238000003786 synthesis reaction Methods 0.000 abstract description 3
- 238000000338 in vitro Methods 0.000 abstract 2
- 238000001727 in vivo Methods 0.000 abstract 2
- 238000011978 dissolution method Methods 0.000 abstract 1
- 238000003756 stirring Methods 0.000 description 10
- 108090001005 Interleukin-6 Proteins 0.000 description 9
- MZOFCQQQCNRIBI-VMXHOPILSA-N (3s)-4-[[(2s)-1-[[(2s)-1-[[(1s)-1-carboxy-2-hydroxyethyl]amino]-4-methyl-1-oxopentan-2-yl]amino]-5-(diaminomethylideneamino)-1-oxopentan-2-yl]amino]-3-[[2-[[(2s)-2,6-diaminohexanoyl]amino]acetyl]amino]-4-oxobutanoic acid Chemical compound OC[C@@H](C(O)=O)NC(=O)[C@H](CC(C)C)NC(=O)[C@H](CCCN=C(N)N)NC(=O)[C@H](CC(O)=O)NC(=O)CNC(=O)[C@@H](N)CCCCN MZOFCQQQCNRIBI-VMXHOPILSA-N 0.000 description 8
- 108060008682 Tumor Necrosis Factor Proteins 0.000 description 8
- 102000000852 Tumor Necrosis Factor-alpha Human genes 0.000 description 8
- 210000002966 serum Anatomy 0.000 description 8
- 239000000243 solution Substances 0.000 description 8
- 241000699670 Mus sp. Species 0.000 description 7
- 206010061218 Inflammation Diseases 0.000 description 6
- 150000001875 compounds Chemical class 0.000 description 6
- 230000001939 inductive effect Effects 0.000 description 6
- 230000004054 inflammatory process Effects 0.000 description 6
- 238000001953 recrystallisation Methods 0.000 description 6
- 238000011160 research Methods 0.000 description 6
- 239000013078 crystal Substances 0.000 description 5
- 239000000706 filtrate Substances 0.000 description 5
- 239000007788 liquid Substances 0.000 description 5
- 238000001819 mass spectrum Methods 0.000 description 5
- 238000001556 precipitation Methods 0.000 description 5
- 239000000047 product Substances 0.000 description 5
- 150000001335 aliphatic alkanes Chemical class 0.000 description 4
- 229960000935 dehydrated alcohol Drugs 0.000 description 4
- UREBDLICKHMUKA-CXSFZGCWSA-N dexamethasone Chemical compound C1CC2=CC(=O)C=C[C@]2(C)[C@]2(F)[C@@H]1[C@@H]1C[C@@H](C)[C@@](C(=O)CO)(O)[C@@]1(C)C[C@@H]2O UREBDLICKHMUKA-CXSFZGCWSA-N 0.000 description 4
- 229960003957 dexamethasone Drugs 0.000 description 4
- 238000010586 diagram Methods 0.000 description 4
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 description 3
- 239000000470 constituent Substances 0.000 description 3
- 229940079593 drug Drugs 0.000 description 3
- 230000000694 effects Effects 0.000 description 3
- 239000007924 injection Substances 0.000 description 3
- 238000002347 injection Methods 0.000 description 3
- 239000002904 solvent Substances 0.000 description 3
- 210000003462 vein Anatomy 0.000 description 3
- 238000002965 ELISA Methods 0.000 description 2
- 241000699666 Mus <mouse, genus> Species 0.000 description 2
- 230000009286 beneficial effect Effects 0.000 description 2
- 210000004369 blood Anatomy 0.000 description 2
- 239000008280 blood Substances 0.000 description 2
- 229960004756 ethanol Drugs 0.000 description 2
- 239000002609 medium Substances 0.000 description 2
- 239000000376 reactant Substances 0.000 description 2
- 230000004044 response Effects 0.000 description 2
- 239000011780 sodium chloride Substances 0.000 description 2
- 230000009897 systematic effect Effects 0.000 description 2
- 238000012360 testing method Methods 0.000 description 2
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 2
- 238000005160 1H NMR spectroscopy Methods 0.000 description 1
- NHBKXEKEPDILRR-UHFFFAOYSA-N 2,3-bis(butanoylsulfanyl)propyl butanoate Chemical compound CCCC(=O)OCC(SC(=O)CCC)CSC(=O)CCC NHBKXEKEPDILRR-UHFFFAOYSA-N 0.000 description 1
- 201000006474 Brain Ischemia Diseases 0.000 description 1
- 206010008120 Cerebral ischaemia Diseases 0.000 description 1
- 241000142975 Cornaceae Species 0.000 description 1
- 241000581650 Ivesia Species 0.000 description 1
- 206010054949 Metaplasia Diseases 0.000 description 1
- 230000009471 action Effects 0.000 description 1
- 239000013543 active substance Substances 0.000 description 1
- 230000003064 anti-oxidating effect Effects 0.000 description 1
- 230000008901 benefit Effects 0.000 description 1
- 244000309464 bull Species 0.000 description 1
- 239000006285 cell suspension Substances 0.000 description 1
- 206010008118 cerebral infarction Diseases 0.000 description 1
- 230000008859 change Effects 0.000 description 1
- 229940126678 chinese medicines Drugs 0.000 description 1
- 238000004440 column chromatography Methods 0.000 description 1
- 208000012839 conversion disease Diseases 0.000 description 1
- 238000001514 detection method Methods 0.000 description 1
- 206010012601 diabetes mellitus Diseases 0.000 description 1
- 230000029087 digestion Effects 0.000 description 1
- 201000010099 disease Diseases 0.000 description 1
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 1
- 239000000428 dust Substances 0.000 description 1
- IDGUHHHQCWSQLU-UHFFFAOYSA-N ethanol;hydrate Chemical compound O.CCO IDGUHHHQCWSQLU-UHFFFAOYSA-N 0.000 description 1
- 238000006266 etherification reaction Methods 0.000 description 1
- -1 ethyl morroniside Chemical compound 0.000 description 1
- 238000009432 framing Methods 0.000 description 1
- 230000004927 fusion Effects 0.000 description 1
- 239000001963 growth medium Substances 0.000 description 1
- 229960003444 immunosuppressant agent Drugs 0.000 description 1
- 230000001861 immunosuppressant effect Effects 0.000 description 1
- 239000003018 immunosuppressive agent Substances 0.000 description 1
- 230000006872 improvement Effects 0.000 description 1
- 239000007928 intraperitoneal injection Substances 0.000 description 1
- 210000004185 liver Anatomy 0.000 description 1
- 238000004519 manufacturing process Methods 0.000 description 1
- 239000000463 material Substances 0.000 description 1
- 230000015689 metaplastic ossification Effects 0.000 description 1
- 239000000203 mixture Substances 0.000 description 1
- 230000004048 modification Effects 0.000 description 1
- 238000012986 modification Methods 0.000 description 1
- 230000000324 neuroprotective effect Effects 0.000 description 1
- 230000000144 pharmacologic effect Effects 0.000 description 1
- 239000013641 positive control Substances 0.000 description 1
- 230000008569 process Effects 0.000 description 1
- 238000012827 research and development Methods 0.000 description 1
- 230000005070 ripening Effects 0.000 description 1
- 238000000926 separation method Methods 0.000 description 1
- 239000012679 serum free medium Substances 0.000 description 1
- 239000006228 supernatant Substances 0.000 description 1
Classifications
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07H—SUGARS; DERIVATIVES THEREOF; NUCLEOSIDES; NUCLEOTIDES; NUCLEIC ACIDS
- C07H17/00—Compounds containing heterocyclic radicals directly attached to hetero atoms of saccharide radicals
- C07H17/04—Heterocyclic radicals containing only oxygen as ring hetero atoms
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07H—SUGARS; DERIVATIVES THEREOF; NUCLEOSIDES; NUCLEOTIDES; NUCLEIC ACIDS
- C07H1/00—Processes for the preparation of sugar derivatives
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- Chemical & Material Sciences (AREA)
- Organic Chemistry (AREA)
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Engineering & Computer Science (AREA)
- Biochemistry (AREA)
- Biotechnology (AREA)
- General Health & Medical Sciences (AREA)
- Genetics & Genomics (AREA)
- Molecular Biology (AREA)
- Medicines Containing Plant Substances (AREA)
- Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
Abstract
The invention discloses a preparation method for 7-O-ethylmorroniside. An optimal 7-O-ethylmorroniside preparation process is screened by a large number of experiments, and comprises parameters such as a dissolution method, a reaction pH value, heating temperature and heating time, an optimal purification method is selected, the whole reaction process is reasonable in design and high in operability, and the 7-O-ethylmorroniside is high in synthesis rate and purity and low in impurity content and cost, and can be industrially produced. A screening result obtained by in-vitro and in-vivo anti-inflammatory experiments further shows that the 7-O-ethylmorroniside has good in-vitro and in-vivo anti-inflammatory effects and has a potential use for preparing an anti-inflammatory medicament.
Description
Technical field
The present invention relates to the preparation method of a kind of compound, be specifically related to the preparation method of 7-O-ethyl morroniside, belong to medical science
Field.
Background technology
Fructus Corni is the drying and ripening sarcocarp of Cornaceae plant Fructus Corni, is one of 40 kinds of extremely conventional rare Chinese medicines.Existing
Generation research shows, iridoid glycoside constituents is the important active substances of Fructus Corni, have antiinflammatory, protect the liver, immunosuppressant etc. is lived
Property.Current pharmaceutical research the most tentatively discloses the activity of the wherein composition such as morroniside, loganin, the new glycosides of Fructus Corni, and
The pharmacological action of other iridoid glycoside constituents is still not clear at present.It is diabetes complicated that literature research shows that morroniside has improvement
Disease acts on alleviating cerebral ischemia etc., has antiinflammatory, antioxidation and neuroprotective isoreactivity.7-O-ethyl morroniside is mountain
A kind of iridoid glycoside constituents (content is about 0.05～0.06mg/g) in Fructus Corni, containing morroniside framing structure, but it is
The no pharmacologically active having morroniside similar yet there are no systematic research.Due to 7-O-ethyl morroniside content in Fructus Corni decoction pieces
Less, separation from decoction pieces, purification can not preferably meet the needs of inside and outside pharmacology activity research.Therefore, present invention research and development
The method of the Synthesis of 7-O-ethyl morroniside, is carried out the inside and outside antiinflammatory action of 7-O-ethyl morroniside on this basis
Systematic research.
Summary of the invention
Goal of the invention: it is an object of the invention to provide the new preparation process of a kind of 7-O-ethyl morroniside, and the present invention investigates 7-O-
The inside and outside antiinflammatory action of ethyl morroniside.
Technical scheme: in order to realize object above, the technical scheme that the present invention takes is:
The preparation method of 7-O-ethyl morroniside, comprises the following steps:
(1) dissolve: taking iridoid glycoside in Fructus Corni Officinalis is raw material, adds raw material weight 10～the ethanol of 20 times of volumes, ultrasonic dissolution；
(2) pH is adjusted: after ultrasonic dissolution, the pH value of regulation solution to 1～3 or addition acetylation reagent；
(3) heating: heating and refluxing extraction 10～20h, filters to obtain extracting solution, is concentrated to give extractum；
(4) purification, crystallization: extractum is dissolved in ethyl acetate, filters, and removes insoluble matter, concentrates, and concentrated solution adds dichloromethane,
Placing crystallization, repeat to crystallize 1～3 time, drying under reduced pressure i.e. obtains 7-O-ethyl morroniside.
Preferably, the preparation method of above-described 7-O-ethyl morroniside, iridoid glycoside in Fructus Corni Officinalis is with mountain Zhu
Cornel is raw material, uses water extracting method to prepare iridoid glycoside in Fructus Corni Officinalis.Preferably, Fructus Corni iridoid
Total glycosides contains percentage by weight 20～40% morroniside and percentage by weight 10～20% loganin.
As more preferably scheme, the preparation method of described 7-O-ethyl morroniside, containing weight in iridoid glycoside in Fructus Corni Officinalis
Amount percentage ratio 40% morroniside and percentage by weight 20% loganin.
Preferably, the preparation method of above-described 7-O-ethyl morroniside, in step (1), ethanol is absolute alcohol；
Supersonic frequency is 20～60kHz, and power is 30kW.The present invention obtains the supersonic frequency of optimal ultrasonic dissolution by great many of experiments screening
Rate and power, be greatly improved the dissolution rate of reactant, it is ensured that reaction conversion ratio.
Preferably, the preparation method of above-described 7-O-ethyl morroniside, the pH value regulator in step (2) is
One in phosphoric acid, hydrochloric acid, sulphuric acid or nitric acid.The present invention is screened by great many of experiments, and regulation reactant pH value is 1～3, can
Improve synthesis rate, improve reaction yield, can make the conversion ratio of 7-O-ethyl morroniside more than 93%.
Preferably, the preparation method of above-described 7-O-ethyl morroniside, 80～100 DEG C of heating in step (3)
Reflux, extract, 16h, the present invention screens different reaction temperatures and response time by great many of experiments, and contrast and experiment shows,
The synthetic ratio that can make 7-O-ethyl morroniside 80～100 DEG C of heating and refluxing extraction 16h is the highest, and impurity is few, and yield is high.
Preferably, the preparation method of above-described 7-O-ethyl morroniside, purification step, the present invention is by a large amount of real
Test the solvent that screening is different, test result indicate that employing acetic acid ethyl dissolution, can effectively remove insoluble matter, and use preferably
Dichloromethane solvent as recrystallization solvent, the 7-O-ethyl morroniside of purity more than 98% can be obtained.The present invention need not
Through the purification step that column chromatography is complicated, faster, work efficiency is higher for purification speed, and low cost is more beneficial for industrialized production.
The present invention is tested by inside and outside antiinflammatory, and result is respectively provided with good antiinflammatory effect with external in showing 7-O-ethyl morroniside body
Really.
Beneficial effect: compared to the prior art the preparation method of the 7-O-ethyl morroniside that the present invention provides has the advantage that
The present invention filters out optimal 7-O-ethyl morroniside preparation technology by great many of experiments, and including dissolving method, regulation is preferably
PH value, heating-up temperature, heat time heating time, response parameter, and preferably went out optimal purification process, and whole reaction process designs
Rationally, workable, 7-O-ethyl morroniside synthetic ratio is high, and impurity is few, and purity is high, low cost, can realize industry metaplasia
Produce.
The present invention, by inside and outside antiinflammatory experiment screening, well resists with external being respectively provided with in proving 7-O-ethyl morroniside body first
Scorching effect, may be used for being prepared as anti-inflammatory drug.
Accompanying drawing explanation
Fig. 1 is the structural representation of 7-O-ethyl morroniside provided by the present invention.
Fig. 2 is the block diagram of the Raw264.7 emiocytosis TNF-α impact that LPS is stimulated by 7-O-ethyl morroniside.
Fig. 3 is the block diagram of the Raw264.7 emiocytosis IL-6 impact that LPS is stimulated by 7-O-ethyl morroniside.
Fig. 4 is the block diagram that LPS inducible system inflammation mice serum IL-6 level is affected by 7-O-ethyl morroniside.
Fig. 5 is the block diagram that LPS inducible system inflammation mice serum TNF-α level is affected by 7-O-ethyl morroniside.
Detailed description of the invention
According to following embodiment, the present invention be may be better understood.But, as it will be easily appreciated by one skilled in the art that embodiment
Described concrete material proportion, process conditions and result thereof are merely to illustrate the present invention, and should be also without limitation on right
The present invention described in detail in claim.
Embodiment 1
The preparation method of 7-O-ethyl morroniside, comprises the following steps:
(1) dissolve: weigh iridoid glycoside in Fructus Corni Officinalis (containing morroniside 40%, loganin 20%) 1kg, add 10L
Dehydrated alcohol, under normal temperature condition, stirring is allowed to be uniformly dispersed；
(2) pH value is adjusted: add appropriate phosphoric acid,diluted regulation pH=3；
(3) reacting by heating: at 80 DEG C of 16h that reflux；Filter to obtain extracting solution, be concentrated to give extractum；
(4) purification, crystallization: extractum adds acetic acid ethyl dissolution, filters away insoluble matter, and filtrate is concentrated into small size, to dense
Adding dichloromethane in contracting liquid to stir evenly to there being Precipitation, place crystallization, crystal with acetic acid ethyl dissolution, adds dichloromethane again
Alkane recrystallization, filters drying under reduced pressure and obtains compound 7-O-ethyl morroniside product 365g, conversion ratio 93%.MS points out gained
Adduct molecule amount: 434g/mol, high resolution mass spectrum molecular formula: C19H30O11。(1H-NMR is consistent with document report, sees
Fortunatus SUNGHWA,H.S.,et al.Iodine-Catalyzed Etherification of Morroniside.Chem.Pharm.
Bull, 2009.57 (1): 112-115.), structural formula such as Fig. 1.
Embodiment 2
The preparation method of 7-O-ethyl morroniside, comprises the following steps:
(1) dissolve: weigh iridoid glycoside in Fructus Corni Officinalis (containing morroniside 40%, loganin 20%) 1kg, put into and extract
Tank, adds 15L dehydrated alcohol, and under normal temperature condition, stirring is allowed to be uniformly dispersed；
(2) pH value is adjusted: add appropriate dilute sulfuric acid regulation PH=2；
(3) reacting by heating: 90 DEG C of backflow 16h；Filter to obtain extracting solution, be concentrated to give extractum；
(4) purification, crystallization: extractum adds acetic acid ethyl dissolution, is filtered to remove insoluble matter, and filtrate is concentrated into small size, to dense
Adding dichloromethane in contracting liquid to stir evenly to there being Precipitation, place crystallization, crystal with acetic acid ethyl dissolution, adds dichloromethane again
Alkane recrystallization, filters drying under reduced pressure and obtains compound 7-O-ethyl morroniside product 390g, conversion ratio 98%.MS points out gained
Adduct molecule amount: 434, high resolution mass spectrum molecular formula: C19H30O11.Structural formula such as Fig. 1.
Embodiment 3
The preparation method of 7-O-ethyl morroniside, comprises the following steps:
(1) dissolve: weigh iridoid glycoside in Fructus Corni Officinalis (containing morroniside 40%, loganin 20%) 1kg, add 20L
Dehydrated alcohol, under normal temperature condition, stirring is allowed to be uniformly dispersed；
(2) pH value is adjusted: add appropriate dilute hydrochloric acid regulation pH=1；
(3) reacting by heating: 100 DEG C of reflux, extract, 16h；Filter to obtain extracting solution, be concentrated to give extractum；
(4) purification, crystallization: extractum adds acetic acid ethyl dissolution, filters away insoluble matter, and filtrate is concentrated into small size, to dense
Adding dichloromethane in contracting liquid to stir evenly to there being Precipitation, place crystallization, crystal with acetic acid ethyl dissolution, adds dichloromethane again
Alkane recrystallization, filters drying under reduced pressure and obtains compound 7-O-ethyl morroniside product 380g, and conversion ratio 95%, MS points out gained
Adduct molecule amount: 434g/mol, high resolution mass spectrum molecular formula: C19H30O11.Structural formula such as Fig. 1.
Embodiment 4
The preparation method of 7-O-ethyl morroniside, comprises the following steps:
(1) dissolve: weigh iridoid glycoside in Fructus Corni Officinalis (containing morroniside 30%, loganin 15%) 1kg, add 10L
Dehydrated alcohol, under normal temperature condition, stirring is allowed to be uniformly dispersed；
(2) pH value is adjusted: add appropriate dust technology regulation pH=1；
(3) reacting by heating: 100 DEG C of backflow 16h；Filter to obtain extracting solution, be concentrated to give extractum；
(4) purification, crystallization: extractum adds acetic acid ethyl dissolution, filters away insoluble matter, and filtrate is concentrated into small size, to dense
Adding dichloromethane in contracting liquid to stir evenly to there being Precipitation, place crystallization, crystal with acetic acid ethyl dissolution, adds dichloromethane again
Alkane recrystallization, filters drying under reduced pressure and obtains 7-O-ethyl morroniside product 375g, conversion ratio 93%.MS prompting gained compound divides
Son amount: 434g/mol, high resolution mass spectrum molecular formula: C19H30O11.Structural formula such as Fig. 1.
Embodiment 5
The preparation method of 7-O-ethyl morroniside, comprises the following steps:
(1) dissolve: weigh iridoid glycoside in Fructus Corni Officinalis (containing morroniside 30%, loganin 20%) 1kg, add 20L without
Water-ethanol, under normal temperature condition, stirring is allowed to be uniformly dispersed；
(2) pH value is adjusted: add appropriate dilute hydrochloric acid regulation pH=1.5；
(3) reacting by heating: 100 DEG C of backflow 16h；Filter to obtain extracting solution, be concentrated to give extractum；
(4) purification, crystallization: extractum adds acetic acid ethyl dissolution, filters away insoluble matter, and filtrate is concentrated into small size, to concentration
Adding dichloromethane in liquid to stir evenly to there being Precipitation, place crystallization, crystal with acetic acid ethyl dissolution, adds dichloromethane again
Recrystallization, filters drying under reduced pressure and obtains 7-O-ethyl morroniside product 392g, conversion ratio 98%.MS prompting gained compound divides
Son amount: 434g/mol, high resolution mass spectrum molecular formula: C19H30O11.Structural formula such as Fig. 1.
Embodiment 6 antiinflammatory is tested
1, extracorporeal anti-inflammatory experimental technique:
Experiment is divided into 5 groups, i.e. Normal group, dexamethasone positive drug group, LPS model group, low dose of 7-O-ethyl morroniside
Amount group (5 μMs), 7-O-ethyl morroniside high dose group (20 μMs).Take the logarithm the RAW264.7 cell of trophophase, digestion
Prepare cell suspension, cell density is adjusted to 5 × 104Individual/ml, takes 1mL and is inoculated in 24 well culture plates, and often group 4 is multiple
Hole, is placed in containing 5%CO2, 37 DEG C of incubators are cultivated, be about 80% to degrees of fusion.
Each group dose regimen is: (1) Normal group, adds entirely without blood serum medium；(2) dexamethasone positive drug group, adds
Enter containing dexamethasone (1 μ g/ml) culture medium；(3) LPS model group, adds entirely without blood serum medium；(4) 7-O-ethyl
Morroniside is low, high dose group: be separately added into the serum-free medium containing 5,20 μMs of 7-O-ethyl morronisides；Each experimental group cell
It is placed in containing 5%CO2, in 37 DEG C of incubators after 30min, in addition to normal group, often group adds LPS to final concentration of 1 μ g/ml.
Draw cell supernatant after continuing to cultivate 24h, use ELISA method detection TNF-α, IL-6.
2, internal antiinflammatory experimental technique:
Experiment packet: take BALB/C mice (male, 10-12 week old) 50, be randomly divided into 5 groups, respectively matched group,
LPS model group, dexamethasone positive controls (1.3mg/kg), 7-O-ethyl morroniside low dose group (10mg/kg), 7-O-
Ethyl morroniside high dose group (30mg/kg) (tail vein injection saline+intraperitoneal injection of saline group), often group 10.
Each group dose regimen is: Normal group and LPS model group mouse tail vein injection normal saline, remaining 3 groups of tail vein is corresponding
Medicine.After 1h, in addition to matched group, often group mouse peritoneal injection LPS (0.8mg/kg).Blood, ice bath 10 to 15 points is taken after 4h
Clock, 8000g is centrifuged 10 minutes and separates serum, uses ELISA method to measure IL-6 and TNF-α.
3, experimental result
3.1 extracorporeal anti-inflammatory experimental results:
As in figure 2 it is shown, the 7-O-ethyl morroniside Raw264.7 emiocytosis TNF-α that LPS is stimulated that provides of the present invention
Impact and as it is shown on figure 3, the present invention 7-O-ethyl morroniside Raw264.7 emiocytosis IL-6 that LPS is stimulated that provides
Impact, compares with normal group, and 7-O-ethyl morroniside can substantially suppress the Raw264.7 emiocytosis TNF-α stimulated due to LPS
And IL-6.
Antiinflammatory experimental result in 3.2 bodies:
As shown in Figure 4, the 7-O-ethyl morroniside that the present invention provides is to LPS inducible system inflammation mice serum IL-6 level
Affect and as it is shown in figure 5, the 7-O-ethyl morroniside of present invention offer is to LPS inducible system inflammation mice serum TNF-α water
Flat impact, during mice vivo medicine-feeding 30mg/kg, 7-O-ethyl morroniside can suppress LPS inducible system inflammation Mouse Blood
Clear IL-6, also can suppress the release of TNF-α, and can suppress LPS inducible system inflammation mice serum IL-6 during 10mg/kg
Release.
Embodiment of above only for technology design and the feature of the present invention are described, its object is to allow person skilled in the art understand
Present invention is also carried out, can not limit the scope of the invention with this, all is done according to spirit of the invention
Equivalence change or modification, all should contain within the scope of the present invention.
Claims (7)
- The preparation method of 1.7-O-ethyl morroniside, it is characterised in that comprise the following steps:(1) dissolve: taking iridoid glycoside in Fructus Corni Officinalis is raw material, adds raw material weight 10～the ethanol of 20 times of volumes, ultrasonic dissolution；(2) pH is adjusted: after ultrasonic dissolution, the pH value of regulation solution to 1～3 or addition acetylation reagent；(3) heating: heating and refluxing extraction 10～20h, filters to obtain extracting solution, is concentrated to give extractum；(4) purification, crystallization: extractum is dissolved in ethyl acetate, filters, and removes insoluble matter, concentrates, and concentrated solution adds dichloromethane, Placing crystallization, repeat to crystallize 1～3 time, drying under reduced pressure i.e. obtains 7-O-ethyl morroniside.
- The preparation method of 7-O-ethyl morroniside the most according to claim 1, it is characterised in that Fructus Corni iridoid Total glycosides contains percentage by weight 20～40% morroniside and percentage by weight 10～20% loganin.
- The preparation method of 7-O-ethyl morroniside the most according to claim 2, it is characterised in that Fructus Corni iridoid Containing percentage by weight 40% morroniside and percentage by weight 20% loganin in total glycosides.
- The preparation method of 7-O-ethyl morroniside the most according to claim 1, it is characterised in that ethanol in step (1) For absolute alcohol；Supersonic frequency is 20～60kHz, and power is 30kW.
- The preparation method of 7-O-ethyl morroniside the most according to claim 1, it is characterised in that the pH in step (2) Value regulator is the one in phosphoric acid, hydrochloric acid, sulphuric acid or nitric acid.
- The preparation method of 7-O-ethyl morroniside the most according to claim 1, it is characterised in that in step (3) 80～100 DEG C of heating and refluxing extraction 16h.
- 7. the 7-O-ethyl morroniside that claim 1 prepares application in preparing anti-inflammatory drug.
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| CN110025626A (en) * | 2019-05-10 | 2019-07-19 | 厦门大学 | Application of the morroniside in preparation treatment chordapsus drug |
| CN111690023A (en) * | 2019-03-13 | 2020-09-22 | 大理大学 | Loganin acetylated derivative iridoid compound and extraction method and application thereof |
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