CN106279081B - Diterpene class compound and preparation method in euphorbia fischeriana - Google Patents
Diterpene class compound and preparation method in euphorbia fischeriana Download PDFInfo
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- CN106279081B CN106279081B CN201610705600.7A CN201610705600A CN106279081B CN 106279081 B CN106279081 B CN 106279081B CN 201610705600 A CN201610705600 A CN 201610705600A CN 106279081 B CN106279081 B CN 106279081B
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- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07D—HETEROCYCLIC COMPOUNDS
- C07D307/00—Heterocyclic compounds containing five-membered rings having one oxygen atom as the only ring hetero atom
- C07D307/77—Heterocyclic compounds containing five-membered rings having one oxygen atom as the only ring hetero atom ortho- or peri-condensed with carbocyclic rings or ring systems
Abstract
The present invention discloses Diterpene class compound and preparation method in a kind of euphorbia fischeriana for inhibiting tulase activity strong, and the concrete structure of compound is as follows：。
Description
Technical field
The present invention relates to from medicinal plant euphorbia fischeriana（Euphorbia fischerianaSteud.）Abietane
Type diterpene compound, it is particularly a kind of to inhibit Diterpene class compound and preparation in the active strong euphorbia fischeriana of tulase
Method.
Background technology
Euphorbia fischeriana is Euphorbiaceae (Euphorbiaceae) euphorbia euphorbia fischeriana（Euphorbia fischerianaSteud.）Root belongs to herbaceos perennial, is mainly distributed on the provinces and regions such as Northeast China, Hebei and Henan
Resourceful, medicine source is sufficient.Euphorbia fischeriana first recorded in《Sheng Nong's herbal classic》, a long time ago, euphorbia fischeriana is just used for civil
Medicine and disease preventing and treating are mainly used for anticancer and treatment tuberculosis.At present, it is reported about the related of compound in euphorbia fischeriana
Road is such as documented in euphorbia fischeriana rich in diterpene-kind compound, structure type mainly have abietane-type, isopimarane type and
Crotons alkane type etc..Listed compound shown below is the Diterpene class of separation and Extraction in the slave euphorbia fischeriana having disclosed
Close object：
。
How from euphorbia fischeriana separation and Extraction goes out the stronger compound of pharmacological activity is the technology solved to be needed to ask at present
Topic.
Invention content
The present invention is to solve the above-mentioned technical problem present in the prior art, and it is strong to provide a kind of inhibition tulase activity
Euphorbia fischeriana in Diterpene class compound and preparation method.
The present invention technical solution be：Diterpene class compound in a kind of euphorbia fischeriana, it is characterised in that
Structural formula is as follows：
The preparation method of Diterpene class compound in above-mentioned euphorbia fischeriana, it is characterised in that in accordance with the following steps into
Row：
A. euphorbia fischeriana dry rhizome is taken, is extracted 3 times with 95% alcohol reflux after crushing, 2 hours every time, merges extraction
Liquid is concentrated under reduced pressure into ethyl alcohol and steams completely, obtains aqueous extract；
B. it is diluted with water extracting solution, then with petroleum ether, ethyl acetate, extracting n-butyl alcohol, respectively obtains petroleum ether extraction
Object, acetic acid ethyl ester extract, n-butyl alcohol extract and water layer crude extract；
C. merge petroleum ether extract and acetic acid ethyl ester extract, pass sequentially through macroreticular resin, silica gel column chromatography, middle pressure
Liquid phantom preparing chromatogram, preparative high performance liquid chromatography are isolated：
The separation condition of medium pressure liquid phantom preparing chromatogram is as follows：
Falsh columns；
Flow velocity：20 ml/min；
Detection wavelength：210nm、280nm；
Column temperature：30℃；
Mobile phase（1）：0-30min methanol：Water=35：65；30-180min methanol：Water=35:65-80:20；Mobile phase
（2）：0-30min methanol：Water=45:55；30-180min methanol：Water=45:55-80:20；Mobile phase（3）：0-30min methanol：
Water=40:60；30-180min methanol：Water=40:60-60:40；
The separation condition of the high performance liquid chromatography is as follows：
Chromatographic column：YMC C18(2)；
Flow velocity：8 ml/min；
Detection wavelength：210nm、280nm；
Column temperature：30℃；
Mobile phase：0-30min methanol：Water=10:90-45:55；30-65min methanol：Water=45:55-90:10；65-
70min methanol：Water=90:10.
Present invention separation and Extraction from euphorbia fischeriana goes out a variety of Diterpene class compounds, is analyzed by spectral data
Identification, in compound there are four types of（A1~A4）For the noval chemical compound having not been reported, and in noval chemical compound there are two types of（A3, A4）Inhibit
The activity of tulase is apparently higher than the prior art, can be as the choice drug of antituberculosis.
Description of the drawings
Diterpenoids from bulbus high-efficient liquid phase chromatogram in Fig. 1 embodiment of the present invention.
Fig. 2 embodiment of the present invention products therefroms are with euphorbia fischeriana to tubercle bacillus inhibiting effect comparison diagram.
Specific embodiment
Embodiment 1：
It carries out in accordance with the following steps：
A. Euphorbiaceae (Euphorbiaceae) euphorbia euphorbia fischeriana is taken（Euphorbia fischerianaSteud.）25 Kg of dry rhizome is extracted 3 times with 95% alcohol reflux after crushing, 2 hours every time, merges extraction
Liquid is concentrated under reduced pressure into ethyl alcohol and steams completely, obtains aqueous extract（LDJ-6）；
B. it is diluted with water extracting solution, then with petroleum ether, ethyl acetate, extracting n-butyl alcohol, respectively obtains petroleum ether extraction
Object, acetic acid ethyl ester extract, n-butyl alcohol extract and water layer crude extract；
C. merge petroleum ether extract and acetic acid ethyl ester extract（Common 1959g）, pass sequentially through macroreticular resin, silicagel column
Chromatography, middle pressure liquid phantom preparing chromatogram（RpC-18）, preparative high performance liquid chromatography（3000 high performance liquid chromatography of Ultimate
Instrument, DAD detectors）Isolated 9 kinds of products（A1~A9）, high-efficient liquid phase chromatogram is as shown in Figure 1.
The separation condition of medium pressure liquid phantom preparing chromatogram is as follows：
Falsh columns；
Flow velocity：20 ml/min；
Detection wavelength：210nm、280nm；
Column temperature：30℃；
Mobile phase（1）：0-30min methanol：Water=35：65；30-180min methanol：Water=35:65-80:20；Mobile phase
（2）：0-30min methanol：Water=45:55；30-180min methanol：Water=45:55-80:20；Mobile phase（3）：0-30min methanol：
Water=40:60；30-180min methanol：Water=40:60-60:40；
The separation condition of the high performance liquid chromatography is as follows：
Chromatographic column：Diamonsil C18(2)（5μm, 250*4.6mm）；
Flow velocity：8 ml/min；
Detection wavelength：210nm、280nm；
Column temperature：30℃；
Mobile phase：0-30min methanol：Water=10:90-45:55；30-65min methanol：Water=45:55-90:10；65-
70min methanol：Water=90:10.
Further by MS, HRMS,1H NMR、13C NMR、HSQC、HMBC、1H-1The spectral techniques pair such as H COSY, NOESY
Separated obtained product carries out Structural Identification.
1. the spectral data of novel compound A1 ~ A4 is as follows：
Mass spectrometry parameters are shown in Table 1.
Table 1
Carbon-13 nmr spectra data（125 MHz）It is shown in Table 2.
Table 2
Hydrogen nuclear magnetic resonance modal data (500 MHz) is shown in Table 3.
Table 3
2. the carbon-13 nmr spectra data of known structure compound A-45 ~ A9 are shown in Table 4.
Finally, it is as follows to extract 9 kinds of isolated product structures by the present invention：
Activity rating is carried out to compound using tubercle bacillus as pattern bacterium.
The present invention is using tubercle bacillus as pattern bacterium, to 9 kinds of gained（A1~A9）Diterpene class compound and radix euphorbiae lantu are big
Halberd aqueous extract（LDJ-6）Tubercle bacillus resistant activity evaluation is carried out.For whole cell primary dcreening operation, 0.1 mg/ml end level items
There is the monomer of inhibition under part, inhibiting effect is shown in Fig. 2.
As can be seen from Figure 2 Diterpene class compound A-13, A4 are apparently higher than the inhibiting effect of tubercle bacillus
Other comparisons.Any dose used suitable for human or animal is made in Diterpene class compound A-13 or A4 that can be of the invention
Type, the content in its pharmaceutical composition are usually 0.1 ~ 95 mass percent.
Claims (2)
1. Diterpene class compound in a kind of euphorbia fischeriana, it is characterised in that structural formula is as follows：
。
2. a kind of preparation method of Diterpene class compound in euphorbia fischeriana as described in claim 1, it is characterised in that
It carries out in accordance with the following steps：
A. euphorbia fischeriana dry rhizome is taken, is extracted 3 times with 95% alcohol reflux after crushing, 2 hours every time, is merged extracting solution, subtract
Pressure is concentrated into ethyl alcohol and steams completely, obtains aqueous extract；
B. it is diluted with water extracting solution, then with petroleum ether, ethyl acetate, extracting n-butyl alcohol, respectively obtains petroleum ether extract, second
Acetoacetic ester extract, n-butyl alcohol extract and water layer crude extract；
C. merge petroleum ether extract and acetic acid ethyl ester extract, pass sequentially through macroreticular resin, silica gel column chromatography, middle hydraulic fluid phase
It is isolated to prepare chromatography, preparative high performance liquid chromatography：
The separation condition of medium pressure liquid phantom preparing chromatogram is as follows：
Flash columns；
Flow velocity：20 ml/min；
Detection wavelength：210 nm、280nm；
Column temperature：30℃；
Mobile phase（1）：0-30min methanol：Water=35：65；30-180min methanol：Water=35:65-80:20；Mobile phase（2）：0-
30min methanol：Water=45:55；30-180min methanol：Water=45:55-80:20；Mobile phase（3）：0-30min methanol：Water=40:
60；30-180min methanol：Water=40:60-60:40；
The separation condition of the high performance liquid chromatography is as follows：
Chromatographic column：Diamonsil C18(2), 5 μm, 250*4.6mm；
Flow velocity：8 ml/min；
Detection wavelength：210 nm、280nm；
Column temperature：30℃；
Mobile phase：0-30min methanol：Water=10:90-45:55；30-65min methanol：Water=45:55-90:10；65-70min first
Alcohol：Water=90:10.
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| Publication number | Priority date | Publication date | Assignee | Title |
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| CN106916162B (en) * | 2017-03-01 | 2018-10-23 | 中日友好医院 | A kind of preparation method of jolkinolide B bulk pharmaceutical chemicals |
| CN108409694B (en) * | 2018-03-13 | 2019-10-29 | 浙江玉安康瑞生物科技有限公司 | Euphorbia ebiacteolata Hayata D and E are used as the purposes of miR-29a inhibitor and apoptosis of mesenchymal stem cell inhibitor |
| CN108840792B (en) * | 2018-04-27 | 2022-07-08 | 兰州大学 | Enantiomeric isopimarane diterpene and preparation method and application thereof |
| CN109456149B (en) * | 2018-11-02 | 2021-06-04 | 大连医科大学 | Aromatic ring group-containing rose alkyl diterpene, preparation method and application in preparation of antituberculosis drugs |
| CN111135181B (en) * | 2019-12-06 | 2021-10-08 | 大连医科大学 | Application of jolkinolide B and analogue thereof in preparation of anti-tuberculosis drugs |
| CN115611720B (en) * | 2022-12-04 | 2023-03-31 | 中日友好医院(中日友好临床医学研究所) | Novel ent-strobane alkane diterpenoid compound, preparation method thereof, pharmaceutical composition and application |
| CN115626932B (en) * | 2022-12-08 | 2023-04-07 | 中日友好医院(中日友好临床医学研究所) | Novel diterpene compound, preparation method thereof, pharmaceutical composition and application thereof in antitumor drugs |
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| CN101829108A (en) * | 2009-03-10 | 2010-09-15 | 湘北威尔曼制药有限公司 | Application of diterpene ginkgolide |
| CN102603765A (en) * | 2012-02-02 | 2012-07-25 | 齐齐哈尔医学院 | Method for extracting active ingredients of Euphorbia fischeriana and diterpenoid medicament prepared by active ingredients |
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| JPH10279574A (en) * | 1997-02-07 | 1998-10-20 | Sagami Chem Res Center | Apoptosis inducer, antitumor agent and basement membrane infiltration inhibitor |
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|---|---|---|---|---|
| CN101829108A (en) * | 2009-03-10 | 2010-09-15 | 湘北威尔曼制药有限公司 | Application of diterpene ginkgolide |
| CN102603765A (en) * | 2012-02-02 | 2012-07-25 | 齐齐哈尔医学院 | Method for extracting active ingredients of Euphorbia fischeriana and diterpenoid medicament prepared by active ingredients |
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| 松香烷内酯型大戟二萜化合物的研究进展;张大永等;《有机化学》;20090415;第29卷(第2期);第188-196页 * |
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