CN107815473A - A kind of diphenyl ether compound and its preparation method and application - Google Patents
A kind of diphenyl ether compound and its preparation method and application Download PDFInfo
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Abstract
The invention discloses a kind of diphenyl ether compound and its preparation method and application.Described diphenyl ether compound is with Fushi's Phomopsis（Phomopsis fukushii）Bacterial strain is fermented, extracts, chromatographs, obtains after purification, and its molecular formula is C18H20O5, structural formula is：The Compound nomenclature is：diphentherB；Fushi's Phomopsis（Phomopsis fukushii）Bacterial strain, bacterial strain deposit number are CCTCC M 2017632, and preservation date is on October 23rd, 2017.Described preparation method is with Fushi's Phomopsis（Phomopsis fukushii）The solid fermentation thing of bacterial strain is raw material, is extracted through organic solvent, silica gel column chromatography, high pressure liquid chromatography separating step.Described application is application of the described diphenyl ether compound in methicillin-resistant staphylococcus aureus resistance medicine is prepared.The compounds of this invention structure is novel, and compound activity is good, there is preferable application prospect.
Description
Technical field
The invention belongs to biological technical field, and in particular to a kind of diphenyl ether compound and preparation method and application.
Background technology
1978, medical worker inspected 200 plants of staphylococcus aureuses by random samples in Shanghai, the methicillin resistance isolated
Staphylococcus aureus (MRSA) is less than 5%.Sample investigation to inpatient in 2009 shows the gold to methicillin resistance
Staphylococcus aureus（MRSA）More than 60%, 40% is increased within year-on-year 2000.The Zhu Baoli of microorganism institute of Chinese Medical Sciences University
And its once the DNA of China, Denmark, Spaniard's gut flora was sequenced by colleague, research display：Chinese's gut flora has
There are more drug resistant genes.Therefore research and development can effectively control the new strategy and novel drugs of drug-resistant bacteria infection, turn into antibiotic and grind
The focus studied carefully.
The content of the invention
The first object of the present invention is to provide a kind of diphenyl ether compound；Second purpose is to provide described hexichol
The preparation method of ether compound；3rd purpose is the application for providing described diphenyl ether compound.
The first object of the present invention is achieved in that described diphenyl ether compound is with Fushi's Phomopsis
（Phomopsis fukushii）Bacterial strain is fermented, extracts, chromatographs, obtains after purification, and its molecular formula is C18H20O5, structural formula
For：
The Compound nomenclature is：diphentherB；
Fushi's Phomopsis（Phomopsis fukushii）Bacterial strain, bacterial strain deposit number are CCTCC M 2017632, are protected
The Tibetan date is on October 23rd, 2017.
The second object of the present invention, which is achieved in that, to be comprised the following steps：
A, solid fermentation：By bacterial strain Fushi's Phomopsis（Phomopsis fukushii）In potato dextrose agar
In in 28 DEG C cultivate 7 days, be inoculated into the 50 ~ 500ml triangular flask of the liquid seed culture medium containing 10 ~ 100ml, at 28 DEG C
Concussion and cultivate obtains Nutrient medium containing bacterium in 5 ~ 10 days under conditions of rotating speed 180rpm rotation concussions, is inoculated into rice solid culture
Culture obtains tunning in base；
B, medicinal extract extracts：The organic solvent ultrasonic extraction 2 ~ 4 times of 1.5 ~ 3 times of solid-liquid volume ratio will be added in tunning, every time 4
~ 8h, merge extract solution, filtering, when being concentrated under reduced pressure into 1/4 ~ 1/2 volume, stand, filter out sediment, be concentrated to give medicinal extract；
C, chloroform-acetone silica gel column chromatography：Silica gel column chromatography on medicinal extract, dress post silica gel is 200 ~ 300 mesh, and dosage is medicinal extract weight
6 ~ 8 times of amounts；It is 1 with volume ratio:0~0:1 chloroform-acetone mixed solvent gradient elution, collect gradient eluent, concentration, warp
TLC is monitored, and merges identical part；
D, petroleum ether-ethyl acetate silica gel column chromatography：By in step C 8:Chloroform-acetone mixed solvent of 2 proportionings elute
To eluent on silica gel column chromatography, dress post silica gel be 200 ~ 300 mesh, dosage is 6 ~ 8 times of eluent weight, with volume ratio be 1:
0~0:1 petroleum ether-ethyl acetate mixed solvent gradient elution, gradient eluent, concentration are collected, is monitored through TLC, is merged identical
Part；
E, high pressure liquid chromatography isolates and purifies：By 7 in D steps:The petroleum ether-ethyl acetate mixed solvent of 3 proportionings is eluted
Obtained eluent is using 40 ~ 60% methanol as mobile phase, using specification as 20mm × 250mm, 5 μm of C18It is stationary phase to prepare post,
Flow velocity is 20ml/min, and UV-detector Detection wavelength is 284nm, each μ L of sample introduction 200, eluent is collected, after repeatedly adding up
It is evaporated, i.e. diphentherB.
The structure of the diphenyl ether compound of the present invention determines by the following method：
The compounds of this invention is light yellow oil；Ultraviolet spectra（Solvent is methanol）,λ max (log ε): 287 (3.60),
206 (4.63)；Infrared spectrum（Pressing potassium bromide troche）ν max: 3450, 2939, 2834, 1686, 1615, 1459, 1347,
1164, 1055, 872 cm-1；HRESIMS shows that the compounds of this invention adds the molecular ion peak m/z339.1216 [M+Na of sodium
]+（C18H20NaO5Calculated value is 339.1208）, with reference to1H and13C H NMR spectroscopies（Fig. 2 and Fig. 3, attribution data are shown in Table 1）Provide it
Molecular formula C18H20O5。
Compound1H H NMR spectroscopies, which are shown in compound, 19 hydrogen signals, including a methyl proton signal H3-7′
（d H 2.27 s）, a methylol proton signal H2-7 （d H4.64 s）, five aryl proton signal [H-2（d H 6.47 s）,
H-4（d H6.57 s）, H-6（d H6.50 s）, H-2'（d H 6. 37 d J=2.2）, H-6'（d H 6.31 d J=2.2], two
Individual methoxyl group signal [OMe-3（d H3.81 s), OMe-3 '（d H3.86 s)], an acetyl protons signal H3-9'（d H
2.58 s）.Compound13C H NMR spectroscopies, which are shown in compound, 18 carbon signals, including a methyl carbon signal C-7 ＇（dC
23.5 q）, a methylol carbon signal C-7（dC68.2 q）, two methoxyl group carbon signal [3-OMe（dC55.9 q）, 3 ＇-
OMe（dC56.2 q）], acetyl group carbon signal [C-8 ＇（dC198.7 s）, C-9 ＇（dC29.8 q）], five sp2Hydridization
Arylmethine carbon signal [C-2（dC104.4 d）, C-4（dC106.6 d）, C-6（dC108.6 d）, C-2 ＇（dC
100.5 d）, C-6 ＇（dC113.6 d）], seven sp2The aryl quaternary carbon signal [C-1 of hydridization（dC155.1 s）, C-3（dC
162.8 s）, C-5（dC143.2 s）, C-1 ＇（dC162.3 s）, C-3 ＇（dC164.1 s）, C-4 ＇（dC 110.4
s）, C-5 ＇（dC142.3s）], wherein four carbon connect oxygen.It is comprehensive1H H NMR spectroscopies and13C H NMR spectroscopy data, illustrate that the compound has
Two phenyl ring are present.There is the quaternary carbon of four company's oxygen in two phenyl ring, in addition to two methoxy substitutions, there should be an oxygen atom
Two phenyl ring of connection form ehter bond.The compound is the derivative of diphenyl ether, has a methyl, methylol, two on phenyl ring
Individual methoxyl group, an acetyl group substitution.Infrared spectrum shows hydroxyl（3450 cm-1）, carbonyl（1686 cm-1）And phenyl
（1615, 1459, 1347cm-1）Absworption peak.HMBC spectrum in H-9 ' (δ H2.58) and C-4 ' (δ C110.4) it is related, card
Real acetyl group is connected to C-4 ' positions, methoxyl group proton OMe (δ H3.81) and C-3 (δ C162.8), methoxyl group proton OMe (δ H
3.86) and C-3 ' (δ C164.1) it is related, it was demonstrated that two methoxyl groups are connected to C-3 and C-3 ' positions, methyl proton H3-7′
(δ H2.27) and C-4 ' (δ C 110.4)/C-5′ (δ C 142.3)/ C-6′ (δ C113.6) it is related, it was demonstrated that methyl is connected to
C-5 ' positions.Methylol proton signal H2-7 （d H4.64）With C-4 (δ C 106.6)/C-5 (δ C 143.2)/ C-6 (δ C
108.6) it is related, it was demonstrated that methylol is connected to C-5 positions.In addition, diagnostic protons signal H-2 (δH 6.47 s), H-4 (δH
6.57 s),H-6 (δH 6.50, s), H-2′ (δH6.37, d, J=2.2) and H-6 ' (δH6.31, d, J=2.2)
The position of above substituent is proved, therefore the structure of compound is confirmed.
The compound of table 11H and13C NMR datas（Solvent is C5D5N）
The third object of the present invention is achieved in that described diphenyl ether compound is preparing anti-methicillin-resistant staphylococcus
Application in staphylococcus medicine.
In recent years, the emphasis of research, which has turned back to find from microorganism, has the new skeleton of different Antibacterial Mechanisms secondary
Metabolite, and plant endogenesis epiphyte metabolite is the most noticeable focus in the field, wherein most often producing antibacterial activity
The monoid of compound is Phomopsis（Phomopsis）, Phoma（Phoma）And Fusarium（Fusarium）.Intend stem
The mould category of point is the one big category in Deuteromycotina, Coelomycetes fungi, and such bacterial strain can produce abundant secondary metabolite,
What Weber etc. isolated from Erythrina crista-galliPhomopsis Sp. one of middle acquisition new antibacterial activity lactone compound
Phomol, the compound carry out biological test to 24 kinds of bacteriums and fungi and all show good antibacterial activity.Horn etc.
Isolated from Salix gracilistylaPhomopsis sp.Bacterial strain, the cytochalasin Alkaloid phomopsichalasin being separated to
Staphylococcus aureus, pseudomonas aeruginosa, Bacillus subtillis, gamboge coccus, Escherichia coli can be suppressed.
Many natural diphenyl ether show good antibacterial activity.The present invention is from one plant of Fushi's Phomopsis
（Phomopsis fukushii）Isolated one new diphenyl ether compound diphenther B, the change in tunning
Compound has significant anti-MRSA activity.
The diphenther B of the present invention are to be separated first, by nuclear magnetic resonance, mass spectrum, infrared spectrum, ultraviolet
Spectroscopic data determines its molecular formula and structure.Its anti-Staphylococcus aureus reference culture is tested with micro broth dilution method
ATCC25923 and methicillin-resistant staphylococcus aureus reference culture ATCC43300 and activity, diphenther B are resistant to
ATCC25923 staphylococcus aureus reference cultures ATCC25923 MIC is 8 μ g/ml, anti-MRSA reference cultures
ATCC43300MIC is 8 μ g/ml, illustrates that the compound has preferably anti-MRSA activity, can be as anti-MRSA lead compound.
Brief description of the drawings
Fig. 1 is the proton nmr spectra of compound（1H NMR）；
Fig. 2 is the carbon-13 nmr spectra of compound（13C NMR）；
Fig. 3 is the HMBC correlation spectrograms of compound.
Embodiment
With reference to embodiment and accompanying drawing, the present invention is further illustrated, but the present invention is not subject in any way
Limitation, based on present invention teach that any conversion or replacement made, belong to protection scope of the present invention.
Diphenyl ether compound of the present invention is with Fushi's Phomopsis（Phomopsis fukushii）Bacterial strain passes through
Fermentation, extraction, chromatograph, obtain after purification, its molecular formula is C18H20O5, structural formula is：
The Compound nomenclature is：diphentherB；
Fushi's Phomopsis（Phomopsis fukushii）Bacterial strain, bacterial strain deposit number are CCTCC M 2017632, are protected
The Tibetan date is on October 23rd, 2017.
The preparation method of diphenyl ether compound of the present invention, comprises the following steps：
A, solid fermentation：By bacterial strain Fushi's Phomopsis（Phomopsis fukushii）In potato dextrose agar
In in 28 DEG C cultivate 7 days, be inoculated into the 50 ~ 500mL triangular flask of the liquid seed culture medium containing 10 ~ 100mL, at 28 DEG C
Concussion and cultivate obtains Nutrient medium containing bacterium in 5 ~ 10 days under conditions of rotating speed 180rpm rotation concussions, is inoculated into rice solid culture
Culture obtains tunning in base；
B, medicinal extract extracts：The organic solvent ultrasonic extraction 2 ~ 4 times of 1.5 ~ 3 times of solid-liquid volume ratio will be added in tunning, every time 4
~ 8h, merge extract solution, filtering, when being concentrated under reduced pressure into 1/4 ~ 1/2 volume, stand, filter out sediment, be concentrated to give medicinal extract；
C, chloroform-acetone silica gel column chromatography：Silica gel column chromatography on medicinal extract, dress post silica gel is 200 ~ 300 mesh, and dosage is medicinal extract weight
6 ~ 8 times of amounts；It is 1 with volume ratio:0~0:1 chloroform-acetone mixed solvent gradient elution, collect gradient eluent, concentration, warp
TLC is monitored, and merges identical part；
D, petroleum ether-ethyl acetate silica gel column chromatography：By in step C 8:Chloroform-acetone mixed solvent of 2 proportionings elute
To eluent on silica gel column chromatography, dress post silica gel be 200 ~ 300 mesh, dosage is 6 ~ 8 times of eluent weight, with volume ratio be 1:
0~0:1 petroleum ether-ethyl acetate mixed solvent gradient elution, gradient eluent, concentration are collected, is monitored through TLC, is merged identical
Part；
E, high pressure liquid chromatography isolates and purifies：By 7 in D steps:The petroleum ether-ethyl acetate mixed solvent of 3 proportionings is eluted
Obtained eluent is using 40 ~ 60% methanol as mobile phase, using specification as 20mm × 250mm, 5 μm of C18It is stationary phase to prepare post,
Flow velocity is 20ml/min, and UV-detector Detection wavelength is 284nm, each μ L of sample introduction 200, eluent is collected, after repeatedly adding up
It is evaporated, i.e. diphentherB.
Potato dextrose agar described in step A is potato 200g/L, glucose 20g/L, agar
20g/L, 6.5,121 DEG C of pH value, 25min sterilizings, it is cooled to 60 DEG C of contra bevels and is prepared.
Liquid seed culture medium described in step A is NaNO32g/L, K2HPO41g/L, MgSO4·7H2O 0.5g/L,
KCL 0.5g/L, FeSO4·7H2O 0.01g/L, glucose 20g/L, pH value 6.5, loading 250mL conical flasks, 100mL/ bottles,
121 DEG C, 25min sterilizes to obtain.
Rice solid medium described in step A is rice 100g, perlite 20g, distilled water 100ml, loads 600mL
Tissue culture bottle, 121 DEG C, 25min sterilizings are prepared.
Organic solvent described in step B is acetone, ethanol or the methanol of volumetric concentration 60 ~ 80%.
Medicinal extract described in step C is before through silica gel column chromatography, in addition to the acetone that medicinal extract is measured with weight than 1.5 ~ 3 times
Dissolving, 1 ~ 3 times of 200 ~ 300 mesh silica gel mixed samples are then weighed with medicinal extract.
Chloroform-acetone mixed solvent volume proportion described in step C is 1:0、 20:1、9:1、 8:2、7:3、3:2、1:
1、1:2、0:1。
Petroleum ether-ethyl acetate mixed solvent volume proportion described in D steps is 1:0、 20:1、9:1、 8:2、7:3、
3:2、1:1、1:2、0:1。
The preparation method of diphenyl ether compound of the present invention, concrete operations are as follows：
（1）Fushi's Phomopsis is placed in inclined-plane solid medium and saved backup, strain transfer will be saved backup to liquid strain
In sub- culture medium, 28 DEG C of cultures 24 h, rotating speed 180r/min, obtain Fushi's Phomopsis seed liquor on shaking table；Fushi is taken to intend stem
The mould seed liquor of point, is accessed in rice solid medium, 25 ~ 35 DEG C by culture medium mass ratio 2 ~ 5%, is fermented 30 ~ 60 days, is sent out
Ferment product.
（2）Ultrasonic extraction：Using 60 ~ 80% acetone as solvent, ultrasonic extraction 3 ~ 5 times, 4 ~ 8h every time, extract solution is merged,
Filtering, is concentrated under reduced pressure into medicinal extract；
（3）Chloroform-acetone silica gel column chromatography：Medicinal extract pure acetone of the weight than 1.5 ~ 3 times of amounts is dissolved, with medicinal extract weight 1 ~ 3
Upper silicagel column after 200-300 mesh silica gel mixed samples again, using volume proportion as 1:0、 20:1、9:1、 8:2、7:3、3:2、1:1、1:
2、0:1 chloroform-methanol gradient wash, eluent and the concentration of each several part are collected respectively, collects washing for each several part respectively
De- liquid simultaneously concentrates；
（4）Petroleum ether-ethyl acetate silica gel column chromatography:By step（2）8:The fraction of 2 parts, measured with weight than 1.5 ~ 3 times
Pure methanol dissolving, with upper silicagel column after the 200-300 mesh silica gel mixed samples of 1 ~ 3 times of medicinal extract weight, using volume proportion as 1:0、 20:
1、9:1、 8:2、7:3、3:2、1:1、1:2、0:1 petroleum ether-ethyl acetate solution gradient washing, collects each several part respectively
Eluent simultaneously concentrates；
（5）High pressure liquid chromatography separates：Take step（3）7:The methanol of 3 fraction 40 ~ 60% is mobile phase, using specification as 20mm ×
250mm, 5 μm of C18It is stationary phase to prepare post, flow velocity 20ml/min, and UV-detector Detection wavelength is 284nm, each sample introduction
200 μ L, eluent is collected, is evaporated after repeatedly adding up, it is described diphenther B to obtain the present invention.
The application of diphenyl ether compound of the present invention is that described diphenyl ether compound is preparing anti-resistance to methoxy
Application in the staphylococcus aureus medicine of XiLin.
With specific embodiment, the present invention will be further described below：
Embodiment 1
（1）Fushi's Phomopsis CCTCC M 2017632 in inclined-plane solid medium will be preserved in and be inoculated in inclined-plane solid culture
Activated in the flat board of base, test it is pure after be inoculated in inclined-plane solid medium and save backup.The inclined-plane solid medium is：Ma Ling
Potato 200g/L, glucose 20g/L, agar 20g/L, 6.5,121 DEG C of pH value, 25min sterilizings, it is cooled to 60 DEG C and is down flat plate and inclined-plane
It is standby；
（2）The Fushi Phomopsis CCTCC M 2017632 saved backup are transferred in liquid seed culture medium, 28 on shaking table
DEG C culture 24 h, rotating speed 180r/min, obtain Fushi's Phomopsis seed liquor.Described liquid seed culture medium is：NaNO3
2g/L, K2HPO41g/L, MgSO4·7H2O 0.5g/L, KCL 0.5g/L, FeSO4·7H2O 0.01g/L, glucose 20g/L,
PH value 6.5, load 250mL conical flasks, 100mL/ bottles, 121 DEG C, 25min sterilizings are standby；
（3）Take step（2）Gained Fushi's Phomopsis seed liquor, accessed by culture medium mass ratio 2% in rice solid medium
Solid fermentation is carried out, fermentation temperature is 25 DEG C, and fermentation in 45 days is completed.Described rice medium is：Rice 100g, perlite
20g, distilled water 100mL, load 600mL tissue culture bottles, 121 DEG C, 25min sterilizings are standby.
Embodiment 2
Embodiment 1 is repeated, there is following difference：
(3) fermentation temperature is 28 DEG C, and fermentation in 40 days is completed.
Embodiment 3
(3) fermentation temperature is 30 DEG C, and fermentation in 30 days is completed.
Embodiment 4
(3) Fushi's Phomopsis seed liquor is accessed in rice solid medium by culture medium mass ratio 5% and carries out solid fermentation,
Fermentation temperature is 28 DEG C, and fermentation in 35 days is completed.
Embodiment 5
（1）The tunning 10kg of Fushi Phomopsis CCTCC M 2017632 are taken, with 80% acetone ultrasonic extraction 4 times, every time
4h, extract solution merges, filtering, is concentrated under reduced pressure into medicinal extract 562g.
（2）Medicinal extract mixes sample with after 1000mL acetone solutions with the silica gel 200g of 200-300 mesh, then with 200-300 mesh silicon
Glue post 1200g dress posts carry out chromatography, use volume proportion as 20:1, 9:1, 8:2 chloroform-acetone solution gradient is washed
It is de-, eluent and the concentration of each several part are collected respectively.
（3）Take（2）Middle chloroform-methanol 8:2 elution concentrating part 56.3g, are dissolved with 120mL pure methanol, with medicinal extract weight
Upper silicagel column after the 200-300 mesh silica gel mixed samples of 1 ~ 3 times of amount, using volume proportion as 1:0、 20:1、9:1、 8:2、7:3、3:2、
1:1、1:2、0:1 petroleum ether-ethyl acetate solution gradient washing, eluent and the concentration of each several part are collected respectively；
（4）Take step（3）7:3 fraction 12.5g, using 56% methanol as mobile phase, using specification as 20mm × 250mm, 5 μm
C18It is stationary phase to prepare post, and flow velocity 20ml/min, UV-detector Detection wavelength is 287nm, each μ L of sample introduction 200, is collected
15.1min chromatographic peak, it is evaporated after repeatedly adding up, it is described diphenther B to obtain the present invention.
Embodiment 6
Embodiment 5 is repeated, there is following difference：
（1）With 70% acetone ultrasonic extraction 4 times, each 5h；The extractum A being concentrated under reduced pressure is 622g.
（4）Chloroform-acetone 7:3 elution fractions are 15.6g, and high performance liquid chromatography point is carried out by mobile phase of 50% methanol
From collecting 13.8min chromatographic peak.
Embodiment 7
Embodiment 5 is repeated, there is following difference：
（1）With 75% acetone ultrasonic extraction 3 times；The extractum A being concentrated under reduced pressure is 577g.
（4）Chloroform-acetone 7:3 elution fractions are 13.3g, using 60% methanol as mobile phase, collect 11.2min chromatogram
Peak.
Embodiment 8
Embodiment 5 is repeated, there is following difference：
（3）Chloroform-methanol 7:3 elution fractions are 11.6g, carry out high performance liquid chromatography separation by mobile phase of 50% methanol, receive
Collect 18.3min chromatographic peak.
Embodiment 9
Compound prepared by Example 5, is light yellow gum thing, assay method is：With nuclear magnetic resonance, with reference to other wave spectrum skills
Art identifies structure.
（1）Ultraviolet spectra（Solvent is methanol）,λ max (log ε): 287 (3.60), 206 (4.63)；
（2）Infrared spectrum（Pressing potassium bromide troche）ν max: 3450, 2939, 2834, 1686, 1615, 1459, 1347,
1164, 1055, 872 cm-1；HRESIMS shows that the compounds of this invention adds the molecular ion peak m/z339.1216 [M+Na of sodium
]+（C18H20NaO5Calculated value is 339.1208）, with reference to1H and13C H NMR spectroscopies（Fig. 2 and Fig. 3, attribution data are shown in Table 1）Provide it
Molecular formula C18H20O5。
Compound1H H NMR spectroscopies, which are shown in compound, 19 hydrogen signals, including a methyl proton signal H3-7′
（d H 2.27 s）, a methylol proton signal H2-7 （d H4.64 s）, five aryl proton signal [H-2（d H 6.47 s）,
H-4（d H6.57 s）, H-6（d H6.50 s）, H-2'（d H 6. 37 d J=2.2）, H-6'（d H 6.31 d J=2.2], two
Individual methoxyl group signal [OMe-3（d H3.81 s), OMe-3 '（d H3.86 s)], an acetyl protons signal H3-9'（d H
2.58 s）.Compound13C H NMR spectroscopies, which are shown in compound, 18 carbon signals, including a methyl carbon signal C-7 ＇（dC
23.5 q）, a methylol carbon signal C-7（dC68.2 q）, two methoxyl group carbon signal [3-OMe（dC55.9 q）, 3 ＇-
OMe（dC56.2 q）], acetyl group carbon signal [C-8 ＇（dC198.7 s）, C-9 ＇（dC29.8 q）], five sp2Hydridization
Arylmethine carbon signal [C-2（dC104.4 d）, C-4（dC106.6 d）, C-6（dC108.6 d）, C-2 ＇（dC
100.5 d）, C-6 ＇（dC113.6 d）], seven sp2The aryl quaternary carbon signal [C-1 of hydridization（dC155.1 s）, C-3（dC
162.8 s）, C-5（dC143.2 s）, C-1 ＇（dC162.3 s）, C-3 ＇（dC164.1 s）, C-4 ＇（dC 110.4
s）, C-5 ＇（dC142.3s）], wherein four carbon connect oxygen.It is comprehensive1H H NMR spectroscopies and13C H NMR spectroscopy data, illustrate that the compound has
Two phenyl ring are present.There is the quaternary carbon of four company's oxygen in two phenyl ring, in addition to two methoxy substitutions, there should be an oxygen atom
Two phenyl ring of connection form ehter bond.The compound is the derivative of diphenyl ether, has a methyl, methylol, two on phenyl ring
Individual methoxyl group, an acetyl group substitution.Infrared spectrum shows hydroxyl（3450 cm-1）, carbonyl（1686 cm-1）And phenyl
（1615, 1459, 1347cm-1）Absworption peak.HMBC spectrum in H-9 ' (δ H2.58) and C-4 ' (δ C110.4) it is related, card
Real acetyl group is connected to C-4 ' positions, methoxyl group proton OMe (δ H3.81) and C-3 (δ C162.8), methoxyl group proton OMe (δ H
3.86) and C-3 ' (δ C164.1) it is related, it was demonstrated that two methoxyl groups are connected to C-3 and C-3 ' positions, methyl proton H3-7′
(δ H2.27) and C-4 ' (δ C 110.4)/C-5′ (δ C 142.3)/ C-6′ (δ C113.6) it is related, it was demonstrated that methyl is connected to
C-5 ' positions.Methylol proton signal H2-7 （d H4.64）With C-4 (δ C 106.6)/C-5 (δ C 143.2)/ C-6 (δ C
108.6) it is related, it was demonstrated that methylol is connected to C-5 positions.In addition, diagnostic protons signal H-2 (δH 6.47 s), H-4 (δH
6.57 s),H-6 (δH 6.50, s), H-2′ (δH6.37, d, J=2.2) and H-6 ' (δH6.31, d, J=2.2)
The position of above substituent is proved, therefore the structure of compound is confirmed.
Embodiment 10
Compound prepared by Example 6,7,8, is yellow jelly, and assay method is same as Example 9, determines to implement
Compound prepared by example 6,7,8 is C18H20O5。
Embodiment 11
Any compound prepared by Example 5 ~ 8 carries out activity assays, and test situation is as follows：
Using micro-broth dilution method diphenther B MIC（ug/mL）Value.
（1）It is prepared by antibacterials and culture medium：Diphenyl ether compound to be measured is dissolved in DMSO, is configured to concentration (2560 μ
G/ml) mother liquor, filtration sterilization are standby.121 DEG C of sterilizing 30min of the MH broth bouillons prepared are standby.
（2）Culture 24h MRSA bacterium colonies are deployed into bacteria suspension of 0.5 Maxwell than turbid standard.Above-mentioned bacterium is hanged with MH meat soups
Liquid carries out 1: 100 dilution, obtains containing about bacterium 1 × 106CFU/mL bacterium solution is standby.
（3）The preparation and bacterium solution inoculation for diluting antibacterials take sterile test tube（13×100mm）13, except the 1st pipe adds
Outside 1.6mlMH meat soups, often pipe adds MH meat soup 1ml for remaining, and antibacterials stoste is added in the 1st pipe（Such as 1280 μ g/ml）
0.4ml is mixed, and is then drawn 1ml to the 2nd and is managed, and draws the pipes of 1ml to the 3rd after mixing again, so continuous doubling dilution to the 11st pipe,
And draw 1ml from the 11st pipe and discard, the 12nd pipe is the growth control of not drug containing.Now each pipe drug concentration be followed successively by 256,
128、64、32、16、8、4、2、1、0.5、0.25μg/ml.Then above-mentioned each 1ml of the inoculum prepared is added in every pipe, the
1 pipe to the 11st pipe drug concentration is respectively 128,64,32,16,8,4,2,1,05,0.25,0.125 μ g/ml, by dilution tube
Culture is transferred to 96 orifice plates, each sample do three it is parallel, DMSO is solvent control, and vancomycin is positive control.
（4）Incubation, which puts 96 orifice plates being inoculated with 37 DEG C of constant incubators, is incubated 24h.
（5）As a result judge：Using the lowest concentration of drug of complete inhibition bacterial growth in aperture as MIC, while solvent control
Interior bacterium suppresses without obvious.
The bacterial strain used in micro broth dilution method includes staphylococcus aureus reference culture ATCC25923, resistance to methoxy
XiLin staphylococcus aureus reference culture ATCC43300.As a result show diphenther B can suppress ATCC25923 and
ATCC43300, staphylococcus aureus reference culture ATCC25923 MIC are 8 μ g/ml, to methicillin-resistant staphylococcus grape
Coccus reference culture ATCC43300MIC is 8 μ g/ml, can be as the guide for developing methicillin-resistant staphylococcus aureus medicine
Compound.
The foregoing is merely illustrative of the preferred embodiments of the present invention, is not intended to limit the invention, all essences in the present invention
Any modification, equivalent substitution and change made within refreshing and principle etc., should be included in the scope of the protection.
Claims (10)
1. a kind of diphenyl ether compound, it is characterised in that described diphenyl ether compound is with Fushi's Phomopsis
（Phomopsis fukushii）Bacterial strain is fermented, extracts, chromatographs, obtains after purification, and its molecular formula is C18H20O5, structural formula
For：
The Compound nomenclature is：diphentherB；
Fushi's Phomopsis（Phomopsis fukushii）Bacterial strain, bacterial strain deposit number are CCTCC M 2017632, are protected
The Tibetan date is on October 23rd, 2017.
2. the preparation method of diphenyl ether compound described in a kind of claim 1, it is characterised in that comprise the following steps：
A, solid fermentation：By bacterial strain Fushi's Phomopsis（Phomopsis fukushii）In potato dextrose agar
In in 28 DEG C cultivate 7 days, be inoculated into the 50 ~ 500ml triangular flask of the liquid seed culture medium containing 10 ~ 100ml, at 28 DEG C
Concussion and cultivate obtains Nutrient medium containing bacterium in 5 ~ 10 days under conditions of rotating speed 180rpm rotation concussions, is inoculated into rice solid culture
Culture obtains tunning in base；
B, medicinal extract extracts：The organic solvent ultrasonic extraction 2 ~ 4 times of 1.5 ~ 3 times of solid-liquid volume ratio will be added in tunning, every time 4
~ 8h, merge extract solution, filtering, when being concentrated under reduced pressure into 1/4 ~ 1/2 volume, stand, filter out sediment, be concentrated to give medicinal extract；
C, chloroform-acetone silica gel column chromatography：Silica gel column chromatography on medicinal extract, dress post silica gel is 200 ~ 300 mesh, and dosage is medicinal extract weight
6 ~ 8 times of amounts；It is 1 with volume ratio:0~0:1 chloroform-acetone mixed solvent gradient elution, collect gradient eluent, concentration, warp
TLC is monitored, and merges identical part；
D, petroleum ether-ethyl acetate silica gel column chromatography：By in step C 8:Chloroform-acetone mixed solvent of 2 proportionings elute
To eluent on silica gel column chromatography, dress post silica gel be 200 ~ 300 mesh, dosage is 6 ~ 8 times of eluent weight, with volume ratio be 1:
0~0:1 petroleum ether-ethyl acetate mixed solvent gradient elution, gradient eluent, concentration are collected, is monitored through TLC, is merged identical
Part；
E, high pressure liquid chromatography isolates and purifies：By 7 in D steps:The petroleum ether-ethyl acetate mixed solvent of 3 proportionings is eluted
Obtained eluent is using 40 ~ 60% methanol as mobile phase, using specification as 20mm × 250mm, 5 μm of C18It is stationary phase to prepare post,
Flow velocity is 20ml/min, and UV-detector Detection wavelength is 284nm, each μ L of sample introduction 200, eluent is collected, after repeatedly adding up
It is evaporated, i.e. diphentherB.
3. preparation method according to claim 2, it is characterised in that the potato dextrose agar culture described in step A
Base is potato 200g/L, glucose 20g/L, agar 20g/L, 6.5,121 DEG C of pH value, 25min sterilizings, be cooled to 60 DEG C it is oblique
Face is prepared.
4. preparation method according to claim 2, it is characterised in that the liquid seed culture medium described in step A is NaNO3
2g/L, K2HPO41g/L, MgSO4·7H2O 0.5g/L, KCL 0.5g/L, FeSO4·7H2O 0.01g/L, glucose 20g/L,
PH value 6.5, load 250mL conical flasks, 100mL/ bottles, 121 DEG C, 25min sterilizes to obtain.
5. preparation method according to claim 2, it is characterised in that the rice solid medium described in step A is rice
100g, perlite 20g, distilled water 100ml, load 600mL tissue culture bottles, 121 DEG C, 25min sterilizings are prepared.
6. preparation method according to claim 2, it is characterised in that the organic solvent described in step B is volumetric concentration 60
~ 80% acetone, ethanol or methanol.
7. preparation method according to claim 2, it is characterised in that the medicinal extract described in step C is through silica gel column chromatography
Before, in addition to the acetone solution that medicinal extract is measured with weight than 1.5 ~ 3 times, 1 ~ 3 times of 200 ~ 300 mesh silica gel are then weighed with medicinal extract and are mixed
Sample.
8. preparation method according to claim 2, it is characterised in that chloroform-acetone mixed solvent body described in step C
Product proportioning is 1:0、 20:1、9:1、 8:2、7:3、3:2、1:1、1:2、0:1.
9. the preparation method according to claim 2 or 6, it is characterised in that the petroleum ether-ethyl acetate described in D steps mixes
Bonding solvent volume proportion is 1:0、 20:1、9:1、 8:2、7:3、3:2、1:1、1:2、0:1.
A kind of 10. application of the diphenyl ether compound described in claim 1, it is characterised in that described diphenyl ether compound
Application in methicillin-resistant staphylococcus aureus resistance medicine is prepared.
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Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2011103193A2 (en) * | 2010-02-16 | 2011-08-25 | Synthezyme Llc | COPOLYESTERS HAVING REPEAT UNITS DERIVED FROM ω-HYDROXY FATTY ACIDS |
| CN103589757A (en) * | 2013-02-04 | 2014-02-19 | 中国海洋大学 | Preparation method for bromo-diphenyl ether derivatives and applications as antibacterial agents |
| CN104045629A (en) * | 2014-06-25 | 2014-09-17 | 中国科学院化学研究所 | Pyridostatin compounds, and preparation method and application thereof |
-
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| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2011103193A2 (en) * | 2010-02-16 | 2011-08-25 | Synthezyme Llc | COPOLYESTERS HAVING REPEAT UNITS DERIVED FROM ω-HYDROXY FATTY ACIDS |
| CN103589757A (en) * | 2013-02-04 | 2014-02-19 | 中国海洋大学 | Preparation method for bromo-diphenyl ether derivatives and applications as antibacterial agents |
| CN104045629A (en) * | 2014-06-25 | 2014-09-17 | 中国科学院化学研究所 | Pyridostatin compounds, and preparation method and application thereof |
Non-Patent Citations (1)
| Title |
|---|
| CHANTRAPROMMA ET AL.: "2-(4-Acetyl-3,5-dihydroxy-2-methylphenoxy)-4,6-dimethoxy-3-methylbenzoic acid", 《ACTA CRYST.》 * |
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| CN112898161B (en) * | 2021-01-25 | 2022-03-25 | 华南农业大学 | 18-hydroxy octadecanoic carbonate containing diphenyl ether unit and preparation method and application thereof |
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