CN101704826A - Chinese medicinal lilac daphne flower bud diterpene ortho-ester compounds - Google Patents
Chinese medicinal lilac daphne flower bud diterpene ortho-ester compounds Download PDFInfo
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Abstract
The invention relates to Chinese medicinal lilac daphne flower bud diterpene ortho-ester compounds. The compounds of the invention comprise compounds of the following similar structures, and have the following expressed structures.
Description
Technical field:
The present invention relates to separating effective ingredient from lilac daphne, particularly relate to the new diterpene ortho-ester compounds of separation from lilac daphne.
Background technology:
Lilac daphne (Daphne genkwa Sieb.et Zucc.) is the thymelaeceae daphne plant, is to be used as medicine with dry flower traditionally.Lilac daphne is distributed widely in some areas of the each province, China Yangtze valley and the Huanghe valley.This medicine is traditional drastically purgating water drug, and is poisonous, and bitter, suffering are warm in nature, return lung, spleen, kidney channel.Function removing fluid-retention by purgation, expelling phlegm for arresting cough, detoxifcation desinsection are used for that oedema turgor, chest abdomen ponding, phlegm and retained fluid gather, the circulation of vital energy in the wrong direction is breathed with cough, difficulty in urination and defecation, controls diseases such as the mange favus of the scalp, pernio outward.Because lilac daphne is a toxic herb, so clinical application generally needs to be used as medicine after concocting.Successive dynasties are many with method processes of preparing Chinese medicine such as stir-fry, wine stir-fry and vinegar systems, to cut down its toxicity, relax the symptom of discharge function and stomachache, but since the Song dynasty, in the concocted specification, vinegar system replaced other concocting methods fully throughout the country.
Chemical constitution study to lilac daphne has obtained the minority diterpene ortho-ester compounds many separation from give birth to lilac daphne, and finds that its toxicity is bigger.We are by carrying out the isolation technique of system, modern times, science to vinegar system lilac daphne, obtained diterpene ortho-ester compounds in 11, wherein 8 kinds is the new compound of not seeing bibliographical information, its toxicity is low, active strong, illustrate that it is the main effective constituent of vinegar system lilac daphne, be expected to be used for antitumor, improve the medicine that chest abdomen ponding, diuresis, treatment constipation, the mange favus of the scalp, pernio, the circulation of vital energy in the wrong direction are breathed with cough.Above composition utilizes the HPLC-MS analytical proof, and also containing of different content in giving birth to lilac daphne also is the effective constituent of giving birth to lilac daphne.
Summary of the invention:
Purpose of the present invention: the new chemical ingredients of open lilac daphne, new reactive monomer compound, new compound of the present invention comprises 8 kinds of compounds of following similar structures.
Lilac daphne ester A has structure shown in the structural formula 1.
Lilac daphne ester B has structure shown in the structural formula 2.
Lilac daphne ester C has structure shown in the structural formula 3.
Lilac daphne ester D has structure shown in the structural formula 4.
Lilac daphne ester E has structure shown in the structural formula 5.
Lilac daphne ester F has structure shown in the structural formula 6.
Lilac daphne ester G has structure shown in the structural formula 7.
Lilac daphne ester H has structure shown in the structural formula 8.
New compound preparation method process following steps of the present invention:
Lilac daphne water or organic solvent extraction, the extracting solution concentrating under reduced pressure, make aqueous solution suspension, extract with sherwood oil, chloroform, ethyl acetate respectively, chloroform extract wherein, be total diterpene ortho ester compounds crude extract, continuing can separate with means such as silica gel column chromatography, polymeric amide, Sephadex LH-20 or HPLC repeatedly obtains above 8 kinds of compounds
Wherein said organic solvent comprises ethanol, methyl alcohol, acetone,
When using silica gel column chromatography,, obtain total diterpene ortho ester compounds with high polar solvent wash-out again with low polar solvent elder generation wash-out decon.Wherein, low polar solvent can be chloroform, different ratios sherwood oil-acetone, different ratios petroleum ether-ethyl acetate, different ratios chloroform-methanol etc.; High polar solvent can be chloroform, different ratios sherwood oil-acetone, different ratios petroleum ether-ethyl acetate, different ratios chloroform-methanol.
The present invention uses our multiple separation means to comprise silica gel column chromatography, macroporous adsorbent resin, polymeric amide, sephadex lh-20 column chromatography and preparative high performance liquid chromatography etc., from the Chinese medicine lilac daphne, separate and obtain 37 compounds, 11 diterpene ortho-ester compounds, wherein 8 is new compound of the present invention.
New compound of the present invention, through preliminary bioactivity research, finding that diterpene ortho-ester compounds of the present invention has antitumor, effect such as improve that chest abdomen ponding, diuresis, treatment constipation, the mange favus of the scalp, pernio, the circulation of vital energy in the wrong direction are breathed with cough, is the basic substance of Chinese medicine lilac daphne performance pharmacological action.Utilize the HPLC-MS analytical proof, in giving birth to lilac daphne, also have the above composition of different content.
The present invention also comprises the analytical procedure of compound of the present invention: analyzing as HPLC, is detector with mass spectrum (MS) detection, ultraviolet (UV) detection, circular dichroism spectrum (CD) detection, light scattering detector etc.Detection method is as follows:
The configuration of reference substance solution: get diterpene ortho-ester compounds chemical reference substance precision and take by weighing in right amount, be mixed with an amount of reference substance solution with methyl alcohol.
The configuration of sample solution: precision takes by weighing the lilac daphne sample and (gives birth to lilac daphne, vinegar system lilac daphne) each is an amount of, with 100ml chloroform (or methyl alcohol, ethyl acetate, acetone, acetonitrile, sherwood oil, organic solvent and solution in different concentration thereof such as hexanaphthene) extract, reclaim extracting solution, methyl alcohol (or chloroform, ethyl acetate, acetone, acetonitrile, sherwood oil, organic solvent and solution in different concentration thereof such as hexanaphthene) dissolving, the little chromatographic column pre-treatment of SPE through filling ODS, 95% acetonitrile (or methyl alcohol, ethyl acetate, acetone, chloroform, sherwood oil, organic solvent and solution in different concentration thereof such as hexanaphthene) washing, merge elutriant, dissolve with methanol, constant volume, filter, promptly.
Testing method: carrying out HPLC and analyze, is detector with mass spectrum (MS) detection, ultraviolet (UV) detection, circular dichroism spectrum (CD) detection, light scattering detector etc.With the chromatographic peak area of each compound in the sample, corresponding chromatographic peak area with the standard control sample according to calibration curve method (or 1 method of external standard, 2 methods of external standard etc.), carries out quantitative analysis, calculates, promptly.
The present invention also comprises the pharmaceutical composition that contains diterpene ortho-ester compounds of the present invention.
The described composition pharmaceutical preparation that can be The compounds of this invention be mixed with the medicine acceptable carrier.
These preparations comprise that general pharmaceutical dosage form is as tablet, capsule, particle, oral liquid, injection etc.
Pharmaceutical composition of the present invention is determined usage and dosage according to patient's situation in use, but obeys every day three times, each 1-20 agent, as: 1-20 bag or grain or sheet, every dose of 1mg-1000mg.
Compound of the present invention find its have antitumor, suppress thromboxane A2/prostaglandin(PG) superoxide (TXA2/PGH2) acceptor, diuresis, promotion intestines peristalsis, increase enteron aisle moisture and quicken defecation, rush down inferior effect.Point out it to can be used for preparation antitumor and treatment medicine such as various ascites pleural fluid and food.
The data of its antitumor action such as following table illustrate that antitumor effective constituent is the ortho ester constituents of The compounds of this invention in the lilac daphne, and wherein The compounds of this invention is stronger than known compound Yuanhuadine activity:
| Compound | To Hela cell medium effective concentration (μ mol/L) |
| ??1 | ??10.2 |
| ??2 | ??11.4 |
| ??3 | ??54.6 |
| ??4 | ??23.8 |
| ??5 | ??28.9 |
| ??6 | ??43.7 |
| ??7 | ??12.6 |
| ??8 | ??6.5 |
| Genkwanin | ??＞1000 |
| Yuanhuadine | ??342.2 |
Data such as following table that it promotes the mouse intestinal wriggling illustrate that the effective constituent of performance discharge function in the lilac daphne is the ortho ester constituents of The compounds of this invention, and wherein The compounds of this invention is stronger than known compound Yuanhuadine activity:
| Compound | Effective dose (mmol/Kg body weight) |
| ??1 | ??5.6 |
| ??2 | ??23.0 |
| ??3 | ??10.4 |
| ??4 | ??7.1 |
| ??5 | ??3.9 |
| ??6 | ??8.5 |
| ??7 | ??12.9 |
| ??8 | ??9.5 |
| Genkwanin | ??＞1000 |
| Yuanhuadine | ??178.2 |
The data such as the following table of its diuresis (mouse) illustrate that the effective constituent of performance diuretic properties in the lilac daphne is the ortho ester constituents of The compounds of this invention, and wherein The compounds of this invention is stronger than known compound Yuanhuadine activity:
| Compound | Effective dose (mmol/Kg body weight) |
| ??1 | ??5.6 |
| ??2 | ??23.0 |
| ??3 | ??10.4 |
| ??4 | ??7.1 |
| ??5 | ??3.9 |
| ??6 | ??8.5 |
| ??7 | ??12.9 |
| ??8 | ??9.5 |
| Genkwanin | ??＞1000 |
| Yuanhuadine | ??178.2 |
Compound of the present invention is compared with the compound of known similar structures, side effect still less, activity is stronger, stability is higher.
Description of drawings:
Fig. 1: preparation flow 1 medicinal material extract is separated
Fig. 2: the separation of each component in preparation flow 2 acetic acid ethyl ester extracts
Fig. 3: the separation of each component in preparation flow 3 chloroform extracts
Fig. 4: the CD of compound 1 and ultraviolet curve
Fig. 5: α in the compound 1, the cotton effect of β-unsaturated cyclic ketones structure
Fig. 6: α in the compound 2, the cotton effect of β-unsaturated cyclic ketones structure
Fig. 7: the CD of compound 3 and ultraviolet curve
Fig. 8: α in the compound 3, the cotton effect of β-unsaturated cyclic ketones structure
Fig. 9: the CD of compound 4 and ultraviolet curve
Figure 10: α in the compound 4, the cotton effect of β-unsaturated cyclic ketones structure
Figure 11: α in the compound 5, the cotton effect of β-unsaturated cyclic ketones structure
Figure 12: α in the compound 6, the cotton effect of β-unsaturated cyclic ketones structure
Figure 13: the CD of compound 7 and ultraviolet curve
Figure 14: α in the compound 7, the cotton effect of β-unsaturated cyclic ketones structure
Figure 15: the CD of compound 8 and ultraviolet curve
Figure 16: α in the compound 8, the cotton effect of β-unsaturated cyclic ketones structure
Figure 17: the octant rule rule of unsaturated cyclic ketones structure in the compound 9
Figure 18: the octant rule rule of unsaturated cyclic ketones structure in the compound 9
Embodiment:
Further specify the present invention by the following examples, but not as limitation of the present invention.
Embodiment 1: the separation of compound
Self-control vinegar system lilac daphne 5kg measures 95% alcohol reflux three times, each 4h with ten times.The extracting solution concentrating under reduced pressure is removed the suspension of making water behind the alcohol, extracts with sherwood oil, chloroform, ethyl acetate respectively, and each extracts three times, reclaims solvent and obtains petroleum ether extract 104.2g, chloroform extract 139.5g, acetic acid ethyl ester extract 73.5g.
1. acetic acid ethyl ester extract
Acetic acid ethyl ester extract 60g utilizes silica gel column chromatography, with chloroform/methanol different ratios wash-out
(1) chloroform/methanol (100: 2) gets flow point 173-253, and is refining with silica gel column chromatography, sherwood oil: ethyl acetate (100: 50) wash-out gets compound 34 (13mg) behind the recrystallization.
(2) chloroform/methanol (100: 7) gets flow point 440-534, adopts open ODS column chromatography, with methanol different ratios wash-out (100: 1～1: 100)
1. 40% methanol-eluted fractions gets cut 26-33, through RPLC preparation (detect wavelength: 380nm), moving phase is 45% methyl alcohol, reclaims solvent after connecing the peak, behind the recrystallization compound 22 (11mg).
2. 50% methanol-eluted fractions gets flow point 50-72, through RPLC preparation (detection wavelength: 380nm), moving phase is 65% methyl alcohol, connects the peak in different retention time, gets compound 21 (800mg), compound 20 (8mg), compound 14 (7mg) after the recovery solvent recrystallization respectively.
2. chloroform layer extract
Chloroform extract 100g utilizes silica gel column chromatography repeatedly, with sherwood oil/acetone different ratios wash-out.
(1) sherwood oil/acetone (100: 1) gets flow point 53-64 and 65-85, flow point 53-64 is through silica gel column chromatography repeatedly, petrol ether/ethyl acetate (30: 1) wash-out, recrystallization gets compound 30 (20mg). and flow point 65-85 is through silica gel column chromatography repeatedly, petrol ether/ethyl acetate (100: 1) wash-out, recrystallization gets compound 32 (12mg).
(2) sherwood oil/acetone (100: 3) flow point 117-129, through silica gel column chromatography repeatedly, petrol ether/ethyl acetate wash-out (8: 2), recrystallization get compound 15 (8mg) successively.
(3) sherwood oil/acetone (100: 7) gets flow point 190-221 and 233, flow point 190-221 is successively through silica gel column chromatography repeatedly, chloroform/ethyl acetate (100: 1) wash-out, get time flow point 89-139, inferior flow point 89-139 is again through the sephadex lh-20 column chromatography, and chloroform/methanol wash-out (8: 2) gets Arius and divides 15-18, Arius divides 15-18 through preparation thin layer, chloroform/ethyl acetate (100: 1) wash-out, and recrystallization gets compound 31 (8mg).Flow point 233 recrystallizations get compound 12 (500mg).
(4) sherwood oil/acetone (100: 10) gets flow point 222-269 and 270-304.
Flow point 222-269 is through the polymeric amide chromatogram, chloroform/methanol wash-out (100: 10).
Get time flow point 4-8 and 9-14 during 1. pure chloroform wash-out, flow point 4-8 gets compound 19 (9mg) through preparation ripple layer chromatography, recrystallization.Flow point 9-14 is through the sephadex lh-20 column chromatography, chloroform/methanol (8: 2) wash-out gets Arius and divides 24-34 and 101-116, both are respectively through RPLC preparation (detection wavelength: 210nm), moving phase is respectively 60% and 65% methyl alcohol, reclaim solvent after connecing the peak, get compound 26 (11mg), compound 29 (13mg) behind the recrystallization.
2. chloroform: get time flow point 24-31 during methyl alcohol (100: 1) wash-out, through silica gel column chromatography repeatedly, chloroform/acetone (100: 1) wash-out, Arius divide 41-65, merge, recrystallization gets compound 24 (9mg).
Flow point 270-304 is through silica gel column chromatography repeatedly, sherwood oil/acetone wash-out.
1. sherwood oil/acetone (100: 5) gets time flow point 93-136, and merging, recrystallization get compound 18 (8mg).
2. sherwood oil/acetone (100: 3) time flow point 24-30, through the RPLC preparation (detect wavelength: 210nm), moving phase is respectively 60% methyl alcohol, connect reclaim behind the peak after the solvent recrystallization compound 26 (11mg).
3. sherwood oil/acetone (100: 10) gets time flow point 179-194, and through the sephadex lh-20 column chromatography, chloroform/methanol wash-out (8: 2), recrystallization get compound 28 (15mg).
(5) sherwood oil/acetone (100: 20) wash-out gets flow point 305-312,313-341.
Flow point 305-312 is through silica gel column chromatography repeatedly, and chloroform/ethyl acetate (100: 1) wash-out gets time flow point 35-49, merges, recrystallization gets compound 13 (400mg).
Flow point 313-341 is through silica gel column chromatography repeatedly, chloroform/eluent ethyl acetate
1. chloroform/ethyl acetate (100: 1) wash-out gets time flow point 75-85, through the RPLC preparation (detect wavelength: 210nm), moving phase is 65% methyl alcohol, connect reclaim behind the peak after the solvent recrystallization compound 16 (8mg) and compound 17 (9mg).
2. chloroform/ethyl acetate (100: 2) wash-out gets time flow point 93-123, through RPLC preparation (detection wavelength: 210nm), moving phase is 35% methyl alcohol, connect the peak in different retention time, get compound 35 (10mg), compound 23 (9mg) and compound 25 (12mg) after the recovery solvent recrystallization.
3. chloroform/ethyl acetate (100: 3) wash-out gets time flow point 150-165, through the sephadex lh-20 column chromatography, and chloroform/methanol (7: 3) wash-out, recrystallization gets compound 33 (14mg).
Flow point 342-379 is through silica gel column chromatography repeatedly, chloroform/ethyl acetate different ratios wash-out
1. chloroform/ethyl acetate (100: 2) wash-out gets time flow point 60-82, inferior flow point 60-82 is through the sephadex lh-20 column chromatography, chloroform/methanol (8: 2) wash-out gets Arius and divides 32-48, Arius divides 32-48 again through RPLC preparation (detection wavelength: 210nm), moving phase is 80% methyl alcohol, connect reclaim behind the peak after the solvent recrystallization compound 11 (9mg).
2. chloroform/ethyl acetate (100: 3) wash-out gets time flow point 83-98, inferior flow point 83-98 is through the sephadex lh-20 column chromatography, chloroform/methanol (8: 2) wash-out gets Arius and divides 12-21, Arius divides 12-21 again through RPLC preparation (detection wavelength: 210nm), moving phase is 80% methyl alcohol, connect the peak in different retention time, get compound 3 (11mg) and compound 5 (20mg) after the recovery solvent recrystallization.
3. chloroform/ethyl acetate (100: 5) wash-out gets time flow point 99-107,108-121,122-144.
Inferior flow point 99-107 is through the sephadex lh-20 column chromatography, wash-out must divide 34-40 and 10-12 by Arius respectively when chloroform/methanol (8: 2) and (6: 1), both are through RPLC preparation (detection wavelength: 210nm), moving phase is respectively 78% and 80% methyl alcohol, connect the peak in different retention time, get compound 9 (25mg), compound 1 (7mg), compound 7 (15mg), compound 4 (8mg) after the recovery solvent recrystallization.
Inferior flow point 108-121 is through the sephadex lh-20 column chromatography, chloroform/methanol (7: 3) wash-out gets Arius and divides 7-14, Arius divides 7-14 again through RPLC preparation (detection wavelength: 210nm), moving phase is 74% methyl alcohol, connect the peak in different retention time, get compound 2 (25mg), compound 6 (8mg), compound 8 (9mg) after the recovery solvent recrystallization.
Inferior flow point 122-144 is through the sephadex lh-20 column chromatography, the chloroform/methanol wash-out gets Arius and divides 22-32, Arius divide 22-32 again through RPLC preparation (detect wavelength: 210nm), moving phase is 75% methyl alcohol, connect reclaim behind the peak after the solvent recrystallization compound 10 (22mg).
Embodiment 2: chemical structure is identified
Utilize 1 dimension, 2 dimension nuclear magnetic resonance spectrums (1D, 2D-NMR), mass spectrum (MS), circular dichroism spectrum (CD) the spectrum means of etc.ing and other physico-chemical processes determined the chemical structure of 35 compounds that separation obtains.Wherein the chemical structure and the identification of means of diterpene ortho ester (comprising new compound) are as follows:
Embodiment 3:HPLC-TOF-MS analyzes
The configuration of reference substance solution: compound 1-11 self-control chemical reference substance precision takes by weighing in right amount, is mixed with the reference substance solution of 5 μ g/ml with methyl alcohol.
The configuration of sample solution: precision takes by weighing each 5g of lilac daphne sample (giving birth to lilac daphne, vinegar system lilac daphne), extracted 3 hours with 100ml chloroform Sha Shi extractor, reclaim chloroform extracted solution, the 10ml dissolve with methanol is through the little chromatographic column pre-treatment of SPE of filling ODS, 95% acetonitrile 20ml washing, merge methyl alcohol and 95% acetonitrile elutriant, dissolve with methanol is settled to the 5ml volumetric flask, 0.2 μ m membrane filtration, promptly.
Testing method: sample introduction 10 μ l, carry out HPLC-TOF-MS and analyze, serve as to select ion to draw chromatographic peak area with the M+H peak of each compound, corresponding chromatographic peak area with the standard control sample according to calibration curve method, carries out quantitative calculation and analysis.
The result shows: give birth in lilac daphne, the vinegar system lilac daphne, all contain the present invention and separate 11 diterpene ortho-ester compounds that obtain.
Embodiment 4: the chemical structure of compound 1 is identified
The white amorphous powder provides peak m/z 577.2050[M+Na among the HR-ESI-MS]
+(calculated value 577.2044) illustrates that 1 molecular weight is 554, infers that its molecular formula is C
30H
34O
10, and calculate 14 degrees of unsaturation are arranged in 1.In IR (KBr) spectrum at 3440cm
-1There is the absorption peak explanation in 1 structure, to have hydroxyl, and at 1707cm
-1Absorption peak the existence of carbonyl has been described.By right
13C-NMR,
1The analysis of H-NMR and hsqc spectrum proves that 1 contains 30 carbon, and 4 methyl, 3 methylene radical, 14 methynes and 9 quaternary carbons are wherein arranged, resolve simultaneously carbon spectrum have in 1 as can be known 1 methylol (δ 65.1, C-20), 4 even the methyne of oxygen (δ 72.0, C-5; δ 64.3, C-7; δ 78.1, C-12; δ 80.7, and C-14), (δ 72.2, C-4 for the quaternary carbon of 4 company's oxygen; δ 60.4, C-6; δ 78.5, C-9; δ 84.1, and C-13), (δ 209.4, C-3 for 2 carbonyl carbon; δ 173.1, C-1 "), and (δ 143.0, C-15 for 1 terminal double link; δ 113.5, C-16) and 1 mono-substituted phenyl. and chemical shift is a carbon on the ortho ester at the quaternary carbon of δ 117.9, and this is the structure fragment of feature the most in the diterpene ortho-ester compounds
[1]By to 1
1The analysis that chemical shift of H-NMR spectrum and proton peak are split the branch situation can further be released, and it is respectively δ 1.87 (3H, s, CH that 2 methyl that are connected on the quaternary carbon are arranged in 1
3-17) and δ 1.79 (3H, s, CH
3-19), 1 methyl δ 1.37 (3H, d, CH that is connected on the tertiary carbon
3-18) and 1 methyl δ 1.11 (3H, t, CH that is connected on the secondary carbon
3-3 "); 4 even the proton on the oxygen methyne be 4.27 respectively (1H, s, H-5), 3.61 (1H, s, H-7), 5.06 (1H, s, H-12) and 4.90 (1H, d, J=2.4Hz, H-14); 1 methylol δ 3.82 (1H, d, J=12.0Hz, H that is connected on the quaternary carbon
a-20), δ 3.96 (1H, d, J=12.0Hz, H
b-20); The proton signal that also has 1 terminal double link is respectively at δ 5.00 (1H, s, H
a-16) and δ 5.03 (1H, s, H
b-16).To compound 1
13C-NMR,
1H-NMR, HSQC and HMBC spectrum are carried out integration analysis, and carbon signal and proton signals that it is all are made ownership, infer that 1 should be the compound of winter daphne type diterpene skeleton, and and document
[2]The existence that has more further proved this skeleton of this compounds spectral data of report.
In the HMBC spectrum, quaternary carbon δ 117.9 (C-1 ') and proton δ 4.90 (H-14) and δ 7.71 (H-3 ', 7 ') there be long-range being correlated with, further confirmed the existence of ortho ester structure, and monosubstituted phenyl is connected on the carbon δ 117.9 (C-1 ') of ortho ester; Show and at δ 2.28 (H-2 "), δ 1.11 (CH at the carbonyl carbon signal of δ 173.1 (C-1 ")
3-3 ") and the proton signal of δ 5.06 (H-12) have long-range relevantly, the structure fragment existence of 1 propionyloxy be described, and this fragment is connected on the C-12 (δ 78.1); In the carbonyl carbon signal demonstration of δ 209.4 (C-3) with at δ 1.79 (CH
3-19) and the proton signal of δ 7.59 (H-1) have long-range relevant, δ 1.79 (CH simultaneously
3-19) also exist long-range relevantly with δ 160.4 (C-1) and δ 136.9 (C-2), release in 1 and have the alpha, beta-unsaturated ketone structure, and methyl is connected on the α position; The proton signal of this external δ 3.61 (H-7) and δ 35.5 (C-8), δ 80.7 (C-14), δ 78.5 (C-9), δ 60.4 (C-6) and δ 65.1 (C-20) have long-range relevant; At proton and the δ 113.5 (C-16) of δ 1.87 (H-17), δ 143.0 (C-15) and δ 84.1 (C-13) have relevant; At proton and the δ 78.1 (C-12) of δ 1.37 (H-18), δ 44.1 (C-11) and δ 78.5 (C-9) also have long-range relevant.In sum, the two dimensional structure of compound 1 is determined.
By to NOESY spectrum and
1The peak type of H-NMR spectrum and the analysis of coupling constant can be derived the relative steric configuration of compound 1.In the NOESY spectrum, proton signal δ 5.06 (H-12) and δ 1.37 (CH are arranged
3-18) reference point illustrates that H-11 and H-12 are transconfigurations, again because
1In the H-NMR spectrum proton δ 5.06 (H-12) be rendered as one unimodal, so the angle of H-11 and H-12 should be 90 °; It is relevant that δ 2.43 (H-11) and δ 3.59 (H-8) present, and releasing H-11, H-8 and H-14 is homonymy on six-ring, and 9,13,14-ortho ester structure is then at opposite side; δ 4.90 (H-14) and δ 3.61 (H-7), δ 5.00 (H-16) and δ 1.87 (CH
3-17) relevant, further confirmed above configuration, and terminal double link structure fragment and H-14, H-7 are at homonymy; And
1Proton δ 3.61 (H-7) presents with unimodal in the H-NMR spectrum, illustrates that the interfacial angle of H-7 and H-8 should be near 90 °.δ 3.92 (H-10) and δ 4.27 (H-5) have reference point, illustrate that H-10 and H-5 are homonymy on seven-membered ring.
In the CD spectrum, compound 1 presents negative cotton effect at the 249nm place, and according to α, the CD rule of β-unsaturated cyclic ketones can be determined 1 absolute configuration
[3]Through the system documentation retrieval, find the new compound of this compound for not appearing in the newspapers, and called after Yuanhuaoate A.
Embodiment 5: the chemical structure of compound 2 is identified
The white amorphous powder provides peak m/z 563.1894[M+Na among the HR-ESI-MS]
+(calculated value 563.1887) illustrates that 2 molecular weight is 540, infers that its molecular formula is C
29H
320
10, and calculate 14 degrees of unsaturation are arranged in 2.Comparative compound 1 and 2
1H-NMR,
13The C-NMR data find that the structure of these two compounds is very similar, and it is that acetoxyl group is connected on the C-12 that institute is not both in the compound 2, but not propionyloxy; In HMBC spectrum, δ 169.6 (C-1 ") and δ 2.01 (H-2 ") δ 5.04 (H-12) has the long-range relevant above deduction that confirmed.By analyze 2 NOESY and
1The H-NMR spectrum finds that its relative steric configuration is identical with 1.In the CD spectrum, compound 2 presents negative cotton effect at the 244nm place, and according to α, the CD rule of β-unsaturated cyclic ketones can be determined 2 absolute configuration
[3]Through the system documentation retrieval, find the new compound of this compound for not appearing in the newspapers, and called after Yuanhuaoate F.
Table 2-2 compound 1
1H-(600MHz in CDCl
3) and
13C-NMR (150MHz in CDCl
3) the spectrum data
Compound 2
1H-(300MHz in CDCl
3) and
13C-NMR (75MHz in CDCl
3) the spectrum data
Embodiment 6: the chemical structure of compound 3 is identified
The white amorphous powder provides peak m/z 625.2046[M+Na among the HR-ESI-MS]
+(calculated value 625.2044) illustrates that 3 molecular weight is 602, infers that its molecular formula is C
34H
34O
10, and calculate 18 degrees of unsaturation are arranged in 3.With compound 3 and compound 2
1H-NMR and
13C-NMR compares, and finds that 3 to 2 have lacked 1 methyl, many monosubstituted phenyls, and in the HMBC spectrum, exist δ 165.4 (C-1 ") and δ 7.90 (H-3 ", 7 "), δ's 5.29 (H-12) is long-range relevant, proves that this monosubstituted phenyl should be connected C-1 " on (δ 165.4); Proton signal H-11 in 3 (δ 2.46) and H-12 (δ 5.29) be all than 2 to low field displacement, and this is to be subjected to the deshield result of influence of phenyl ring, further confirms the existence of this structure fragment.Reach by NOESY 3
1The analysis of H-NMR spectrum finds that its relative steric configuration is identical with 2.But in the CD spectrum, compound 3 but presents positive cotton effect at the 240nm place, and according to α, the CD rule of β-unsaturated cyclic ketones can be determined 3 absolute configuration, can determine that this compound is genkwadaphnin (genkwadaphnin).
Embodiment 7: the chemical structure of compound 4 is identified
The white amorphous powder provides peak m/z 643.2151[M+Na among the HR-ESI-MS]
+(calculated value 643.2149) illustrates that 4 molecular weight is 620, infers that its molecular formula is C
34H
36O
11, and calculate 17 degrees of unsaturation are arranged in 4.To compound 4 and compound 3
1H-NMR and
13C-NMR spectrum data compare analysis, find that 4 also is diterpene ortho-ester compounds, and carbon number are identical with 3, and two monosubstituted phenyls are all arranged; Be H-7 (δ 4.56) proton signal and C-6 (δ 76.4), C-7 (δ 80.2) and C-20 carbon signals such as (δ 70.0) all to a low significantly displacement, illustrate 6, and the 7-epoxy construction has not existed, and the substitute is 6, the 7-dihydroxyl.
1δ 4.56 (H-7) presents unimodal in the H-NMR spectrum, the interfacial angle that H-7 and H-8 are described should be 90 °, and δ 3.73 (H-20) has reference point with δ 4.15 (H-5), δ 4.56 (H-7) in NOESY spectrum, satisfy simultaneously at these 2 and have only when H-5, methylol and 7-OH are homonymy on seven-membered ring and just can accomplish.By right
1H-NMR,
13C-NMR, HSQC, HMBC and the analysis-by-synthesis of NOESY spectrum can be determined 4 two dimensional structure and relative steric configuration.In the CD spectrum, compound 4 presents positive cotton effect at the 226nm place, and according to α, the CD rule of β-unsaturated cyclic ketones can be determined 4 absolute configuration
[3]Through the system documentation retrieval, find the new compound of this compound for not appearing in the newspapers, and called after Yuanhuaoate B.
Table 2-3 compound 3 and 4
1H-(600MHz in CDCl
3) and
13C-NMR (150MHz in CDCl
3) the spectrum data
3??????????????????????????????????????????????4
δ(C)??δ(H)(J?in?Hz)???????????????????????????δ(C)??δ(H)(J?in?Hz)
1??????160.2??7.58(1H，brs)?????????????????????????????159.6?7.65(1H，brs)
2??????136.9????????????????????????????????????????????137.2
3??????209.3????????????????????????????????????????????208.9
4??????72.2?????????????????????????????????????????????75.2
5??????71.6???4.21(1H，s)???????????????????????????????73.7??4.15(1H，s)
6??????60.7?????????????????????????????????????????????76.4
7??????64.0???3.68(1H，s)???????????????????????????????80.2??4.56(1H，s)
8??????35.8???3.71(1H，d，J＝2.5)???????????????????????35.9??3.52(1H，d，J＝2.4)
9??????78.6??????????????????????????????????????????????79.1
10???????47.4????3.95(1H，brs)???????????????50.5????4.00(1H，brs)
11???????44.1????2.64(1H，q，J＝7.3)?????????43.9????2.98(1H，q，J＝7.8)
12???????78.9????5.29(1H，s)?????????????????78.1????5.35(1H，s)
13???????84.3????????????????????????????????85.0
14???????80.7????5.04(1H，d，J＝2.5)?????????83.1????5.10(1H，d，J＝2.4)
15???????142.9???????????????????????????????142.5
16???????113.8???5.05(1H，s，H
a)?????????????114.0???5.01(1H，s，H
a)
5.07(1H，s，H
b)?????????????????????5.06(1H，s，H
b)
17???????18.8????1.91(3H，s)?????????????????18.8????1.90(3H，s)
18???????18.4????1.45(3H，d，J＝7.3)?????????18.0????1.44(3H，d，J＝7.2)
19???????9.8?????1.76(3H，brs)???????????????9.9?????1.80(3H，brs)
20???????64.8????3.84(1H，d，J＝12.6，H
a)????70.0????3.73(1H，d，J＝11.4，H
a)
3.94(1H，d，J＝12.6，H
b)????????????4.10(1H，d，J＝11.4，H
b)
1′??????117.9???????????????????????????????117.9
2′??????135.2???????????????????????????????134.8
3′，7′?126.0???7.75(1H，m)?????????????????125.8???7.68(2H，m)
4′，6′?128.6???7.40(1H，m)?????????????????128.5???7.44(2H，m)
5′??????129.7???7.42(1H，m)?????????????????130.0???7.40(1H，m)
1″??????165.4???????????????????????????????165.0
2″??????129.7???????????????????????????????128.5
3″，7″?129.4???7.90(1H，m)?????????????????129.6???7.95(1H，m)
4″，6″?128.1???7.45(1H，m)?????????????????128.2???7.42(1H，m)
5″??????133.3???7.55(1H，m)?????????????????133.3???7.55(1H，m)
Embodiment 8: the chemical structure of compound 5 is identified
The white amorphous powder provides peak m/z671.2833[M+Na among the HR-ESI-MS]
+(calculated value 671.2826) illustrates that 5 molecular weight is 648, infers that its molecular formula is C
37H
44O
10, and calculate 16 degrees of unsaturation are arranged in 5.Comparative compound 5 and 3
1H-NMR,
13After the C-NMR spectrum data, think that 5 also is diterpene ortho-ester compounds, and find that 5 lack a monosubstituted phenyl than 3; And 5 aliphatic carbon and 4 double key carbons have appearred, be respectively δ 122.2 (C-2 '), δ 135.0 (C-3 '), δ 128.5 (C-4 '), δ 139.3 (C-5 '), δ 32.6 (C-6 '), δ 28.6 (C-7 '), δ 31.2 (C-8 '), δ 22.4 (C-9 ') and δ 14.0 (C-10 '); In the HMBC spectrum, δ 1.29 (H-8 ') has long-range relevant with δ 28.6 (C-7 '), δ 22.4 (C-9 ') and δ 14.0 (C-10 '), δ 2.11 (H-6 ') has long-range relevant with δ 28.6 (C-7 '), δ 31.2 (C-8 '), δ 128.5 (C-4 ') and δ 139.3 (C-5 '), release 1 five carbocyclic aliphatic chain of existence in 5, and this aliphatic chain should be connected on two keys; δ 6.06 (H-4 ') and δ 32.6 (C-6 '), δ 122.2 (C-2 ') and δ 135.0 (C-3 ') also exist long-range relevant in the HMBC spectrum, have more proved conclusively the existence of above structure fragment, and illustrate that two pairs of keys should link together; Further analysis 5
1The H-NMR spectrum, 5.69 (H-2 ') are 15.6Hz with the coupling constant of δ 6.70 (H-3 '), δ 6.06 (H-4 ') is 15.0Hz with the coupling constant of δ 5.87 (H-5 '), illustrates that two two keys are trans double bond.Again because in HMBC spectrum, it is long-range relevant that δ 117.0 (C-1 ') and δ 5.69 (H-2 '), δ 6.70 (H-3 ') and δ 4.90 (H-14) have, and can release on the carbon (δ 117.0, C-1 ') that this long-chain should be connected ortho ester.With 5 and 3
1H-NMR and NOESY spectrum compare analysis, find that their diterpene ortho ester skeleton has identical relative steric configuration.But in the CD spectrum, what compound 5 presented at the 226nm place is negative cotton effect, and according to α, the CD rule of β-unsaturated cyclic ketones can be determined 5 absolute configuration
[3]Through the system documentation retrieval, find the new compound of this compound for not appearing in the newspapers, and called after YuanhuaoateD.
Embodiment 9: the chemical structure of compound 6 is identified
The white amorphous powder provides peak m/z689.2939[M+Na among the HR-ESI-MS]
+(calculated value 689.2932) illustrates that 6 molecular weight is 666, infers that its molecular formula is C
37H
46O
11, and calculate 15 degrees of unsaturation are arranged in 6.Comparative compound 6 and 5
13C-NMR composes data, find that the carbon geochemistry change in displacement on the six-ring is very big, at δ 75.4 (C-9), the carbon signal of δ 75.9 (C-13) and δ 73.0 (C-14) all than 5 to high field displacement, and an ester carbonyl group δ 166.9 (C-1 ') appears, ortho ester functional group open loop in 6 that this explanation exists in 5 has become two hydroxyls and a 2E, and 4E-decadienoyl can determine 6 two dimensional structure thus.And 6 relative steric configuration and 5 relatively also just six-ring change, 6
1Have δ 5.19 (H-12) to present doublet in the H-NMR spectrum, and the coupling constant of δ 5.19 (H-12) and δ 2.50 (H-11) is 3.6Hz, illustrates that the interfacial angle between H-11 and the H-12 no longer is 90 °; Again because of δ 5.19 (H-12) and δ 1.88 (CH in the NOESY spectrum
3-17) reference point is arranged, satisfy simultaneously at these 2 and have only the homonymy at six-ring as 9-OH, 13-OH, and H-12 and 2E, 4E-decadienoyl just can accomplish when opposite side.In the CD spectrum, compound 6 presents negative cotton effect at the 244nm place, and according to α, the CD rule of β-unsaturated cyclic ketones can be determined 6 absolute configuration
[3]Through the system documentation retrieval, find the new compound of this compound for not appearing in the newspapers, and called after Yuanhuaoate E.
Table 2-4 compound 5 and 6
1H-(600MHz in CDCl
3) and
13C-NMR (150MHz in CDCl
3) the spectrum data
Embodiment 10: the chemical structure of compound 7 is identified
The white amorphous powder provides peak m/z 609.2673[M+Na among the HR-ESI-MS]
+(calculated value 609.2670) illustrates that 7 molecular weight is 586, infers that its molecular formula is C
32H
42O
10, and calculate 12 degrees of unsaturation are arranged in 7.With compound 7 and 5
1H-NMR,
13C-NMR spectrum data compare analysis, find that 7 are lacked 1 monosubstituted phenyl than 5, and many 1 methyl.In the HMBC spectrum, δ 169.6 (C-1 ") and δ 1.99 (CH
3-2 ") have long-range relevantly, confirmed that further it is acetoxyl group that C-12 in 7 goes up what connect with δ 4.97 (H-12).By to 7
1The integration analysis of H-NMR, NOESY spectrum finds that 7 and 5 have identical relative steric configuration.But in the CD spectrum, compound 7 but presents positive cotton effect at the 230nm place, and according to α, the CD rule of β-unsaturated cyclic ketones can be determined 7 absolute configuration
[3]In sum, can determine that this compound is lilac daphne ester (Yuanhuadin).
Embodiment 11: the chemical structure of compound 8 is identified
The white amorphous powder provides peak m/z 627.2777[M+Na among the HR-ESI-MS]
+(calculated value 627.2775) illustrates that 8 molecular weight is 604, infers that its molecular formula is C
32H
44O
11, and calculate 11 degrees of unsaturation are arranged in 8.Compound 8
1H-NMR,
13C-NMR spectrum data are compared with 6, and many 1 methyl have lacked 1 monosubstituted phenyl; Equally, δ 170.1 (C-1 ") and δ 2.00 (CH in the HMBC spectrum
3-2 "), δ 4.88 (H-12) long-range relevant, also proved in 8 it is that 1 acetoxyl group is connected on the C-12.Analysis-by-synthesis
1H-NMR, NOESY spectrum finds that 8 and 6 have identical relative steric configuration. in the CD spectrum, 8 present at the 236nm place and to bear the cotton effect, and according to α, the CD rule of β-unsaturated cyclic ketones can be determined 8 absolute configuration
[3]Through the system documentation retrieval, find the new compound of this compound for not appearing in the newspapers, and called after YuanhuaoateC.
Table 2-5 compound 7 and 8
1H-(600MHz in CDCl
3) and
13C-NMR (150MHz in CDCl
3) the spectrum data
Embodiment 12: the chemical structure of compound 9 is identified
The white amorphous powder provides peak m/z 627.2207[M+Na among the HR-ESI-MS]
+(calculated value 627.2200) illustrates that 9 molecular weight is 604, infers that its molecular formula is C
34H
36O
10, and calculate 17 degrees of unsaturation are arranged in 9.Comparison 9 and 3
13C-NMR composes data, finds that 9 to 3 lack 2 sp
2The hydridization carbon atom be the substitute is 2 sp
3(δ 33.4, C-1 for the hydridization carbon atom; δ 42.8, C-2); And to low field displacement, illustrate that the structure fragment of alpha, beta-unsaturated ketone does not exist in 9 at the carbonyl carbon signal of δ 220.2 (C-3) the carbonyl carbon signal in than 3.
1In the H-NMR spectrum, δ 1.12 (CH
3-19) proton signal is than the CH in 3
3-19 signals are to high field displacement; δ 2.28 (H-2) and δ 33.4 (C-1), δ 12.3 (CH
3-19) in the HMBC spectrum, have long-range relevant with δ 220.2 (C-3), all further confirmed above deduction, therefore 9 two dimensional structure is determined. in the NOESY spectrum, δ 3.05 (H-10) and δ 2.28 (H-2) have reference point, H-10 and H-2 are described at pentacyclic homonymy, and CH
3-19 at opposite side; Conscientiously resolved 9 NOESY with
1The H-NMR spectrum, the relative steric configuration of 9 rest parts is identical with 3 as can be known.In CD spectrum, 9 present positive cotton effect at the 309nm place, can determine 9 absolute configuration according to " octant rule " rule of saturated cyclic ketones
[4]Through the system documentation retrieval, find the new compound of this compound for not appearing in the newspapers, and called after Yuanhuaoate G.
Embodiment 13: the chemical structure of compound 10 is identified
The white amorphous powder provides peak m/z 565.2052[M+Na among the HR-ESI-MS]
+(calculated value 565.2044) illustrates that 10 molecular weight is 542, infers that its molecular formula is C
29H
34O
10, and calculate 13 degrees of unsaturation are arranged in 10.With compound 10 and 9
13C-NMR,
1The H-NMR spectrum compares analysis, finds more than 10 to 91 methyl, and few 1 monosubstituted phenyl.In the HMBC spectrum, δ 169.7 (C-1 ") and δ 2.02 (CH
3-2 ") δ 5.15 (H-12) have long-range relevant, methyl substituted be described in 10 phenyl, so to be connected on the C-12 be 1 acetoxyl group.Analysis 10
1H-NMR and NOESY spectrum, 10 and 9 have identical relative steric configuration as can be known.In CD spectrum, 10 present positive cotton effect at the 299nm place, can determine 10 absolute configuration according to " octant rule " rule of saturated cyclic ketones
[4]Through the system documentation retrieval, find the new compound of this compound for not appearing in the newspapers, and called after Yuanhuaoate H.
Table 2-6 compound 9 and 10
1H-(600MHz in CDCl
3) and
13C-NMR (150MHz in CDCl
3) the spectrum data
Embodiment 14: the chemical structure of compound 11 is identified
The white amorphous powder provides peak m/z 631.2518[M+Na among the HR-ESI-MS]
+(calculated value 631.2513) illustrates that 11 molecular weight is 608, infers that its molecular formula is C
34H
40O
10, and calculate 15 degrees of unsaturation are arranged in 11.Comparative analysis compound 11 and 9
13C-NMR,
1H-NMR composes data, finds that 11 still is diterpene ortho-ester compounds, the quaternary carbon of δ 117.1 (C-1 ') and δ 4.58 (H-14) and δ 7.69 (H-3 ', 7 ') long-range being correlated with of demonstration further confirmed the existence of ortho ester group in the HMBC spectrum.Difference is, do not have ketone carbonyl signal in 11, the substitute is 1 sp
2Company's oxygen carbon of hydridization (δ 76.7, C-3), and in HMBC spectrum δ 76.7 (C-3) and δ 1.69 (H-1), δ 4.31 (H-3) and δ 13.1 (CH
3-19) long-range being correlated with all arranged, further confirmed such variation; All to low field displacement, illustrate 6 at the carbon signal of δ 77.5 (C-6) and δ 76.3 (C-7), 7-epoxy construction fragment open loop becomes 6,7-dihydroxyl structure fragment; In the HMBC spectrum; ester carbonyl group δ 167.1 (C-1 ") and δ 8.08 (H-3 "; 7 ") show long-range relevant; benzoyl group existence still be described, and δ 4.90 (H-20) (this benzoyl group of long-range relevant prompting of C-1 "), δ 76.3 (C-7) and δ 77.5 (C-6) is connected on the hydroxyl of C-20 with δ 167.1.In composing by analysis HMBC again other are correlated with and have finally been determined 11 two dimensional structure.In the NOESY spectrum, δ 1.74 (H-2) and δ 2.76 (H-10) and δ 4.31 (H-3) have reference point, H-2, H-3 and H-10 are described at pentacyclic homonymy, and CH
3-19 with OH-3 then at opposite side; Analysis-by-synthesis
1H-NMR and NOESY spectrum confirm that the relative steric configuration of rest part is identical with compound 4.According to single-substituted ring
1L
bThe CD rule of band
[5], promptly in the CD spectrum, present negative cotton effect, so C-1 ' should be the S configuration in the 260nm place because of 11.In sum, can determine that this compound is Genkwanine H.
Embodiment 15: preparation of drug combination
Any one of The compounds of this invention 1-8 and the mixing of medicine acceptable carrier are prepared into tablet with the technology of pharmaceutics routine techniques, capsule, particle, oral liquid, preparations such as injection.
Embodiment 16:
In the prior art analogue as: Yuanhuadine and genkwanin and The compounds of this invention carry out pharmacology and toxicity relatively.The compounds of this invention is better than the prior art compound as a result.
| Compound | To Hela cell medium effective concentration (μ mol/L) |
| ??1 | ??10.2 |
| ??2 | ??11.4 |
| ??3 | ??54.6 |
| ??4 | ??23.8 |
| ??5 | ??28.9 |
| ??6 | ??43.7 |
| ??7 | ??12.6 |
| ??8 | ??6.5 |
| Genkwanin | ??＞1000 |
| Yuanhuadine | ??342.2 |
Data such as following table that it promotes the mouse intestinal wriggling illustrate that the effective constituent of performance discharge function in the lilac daphne is the ortho ester constituents of The compounds of this invention, and wherein The compounds of this invention is stronger than known compound Yuanhuadine activity:
| Compound | Effective dose (mmol/Kg body weight) |
| ??1 | ??5.6 |
| ??2 | ??23.0 |
| ??3 | ??10.4 |
| ??4 | ??7.1 |
| ??5 | ??3.9 |
| ??6 | ??8.5 |
| ??7 | ??12.9 |
| ??8 | ??9.5 |
| Genkwanin | ??＞1000 |
| Compound | Effective dose (mmol/Kg body weight) |
| Yuanhuadine | ??178.2 |
The data such as the following table of its diuresis (mouse) illustrate that the effective constituent of performance diuretic properties in the lilac daphne is the ortho ester constituents of The compounds of this invention, and wherein The compounds of this invention is stronger than known compound Yuanhuadine activity:
| Compound | Effective dose (mmol/Kg body weight) |
| ??1 | ??5.6 |
| ??2 | ??23.0 |
| ??3 | ??10.4 |
| ??4 | ??7.1 |
| ??5 | ??3.9 |
| ??6 | ??8.5 |
| ??7 | ??12.9 |
| ??8 | ??9.5 |
| Genkwanin | ??＞1000 |
| Yuanhuadine | ??178.2 |
The mouse toxicity test-results illustrates that the lilac daphne The compounds of this invention is more weak than known compound Yuanhuadine toxicity; Though and genkwanin toxicity is little, it is non-active compound also, thereby the advantage of The compounds of this invention has been described:
| Compound | Medium lethal dose intravenous injection (mmol/Kg body weight) |
| ??1 | ??209.5 |
| ??2 | ??127.5 |
| ??3 | ??118.4 |
| Compound | Medium lethal dose intravenous injection (mmol/Kg body weight) |
| ??4 | ??122.0 |
| ??5 | ??167.8 |
| ??6 | ??98.7 |
| ??7 | ??121.4 |
| ??8 | ??67.2 |
| Genkwanin (flavonoid that from lilac daphne, extracts) | ??＞1000 |
| Yuanhuadine (similar compound belongs to diterpene ortho-ester) | ??356.1 |
Claims (11)
1. a class is separated the diterpene ortho-ester compounds that obtains from the Chinese medicine lilac daphne, comprises lilac daphne ester A-H, and wherein lilac daphne ester A has structure shown in the structural formula 1.
Structural formula 1
2. diterpene ortho-ester compounds: lilac daphne ester B has structure shown in the structural formula 2.
Structural formula 2
3. diterpene ortho-ester compounds: lilac daphne ester C has structure shown in the structural formula 3.
Structural formula 3
4. diterpene ortho-ester compounds: lilac daphne ester D has structure shown in the structural formula 4.
Structural formula 4
5. diterpene ortho-ester compounds: lilac daphne ester E has structure shown in the structural formula 5.
Structural formula 5
6. diterpene ortho-ester compounds: lilac daphne ester F has structure shown in the structural formula 6.
Structural formula 6
7. diterpene ortho-ester compounds: lilac daphne ester G has structure shown in the structural formula 7.
Structural formula 7
8. diterpene ortho-ester compounds: lilac daphne ester H has structure shown in the structural formula 8.
Structural formula 8
Among the claim 1-8 any one diterpene ortho-ester compounds antitumor in preparation, improve the application in the medicine that chest abdomen ponding, diuresis and constipation, the mange favus of the scalp, pernio, the circulation of vital energy in the wrong direction breath with cough.
10. the pharmaceutical composition that contains any one diterpene ortho-ester compounds among the claim 1-8.
11. the preparation method of any one diterpene ortho-ester compounds among the claim 1-8, it is characterized in that, process following steps: the organic solvent extraction of lilac daphne water or different concns, the extracting solution concentrating under reduced pressure, make aqueous solution suspension, use sherwood oil respectively, chloroform, ethyl acetate extracts, chloroform extract, be total diterpene ortho ester compounds crude extract, continue and use silica gel column chromatography repeatedly, polymeric amide, Sephadex LH-20, can separate with means such as HPLC and to obtain above compound, wherein organic solvent comprises ethanol, methyl alcohol, acetone, during silica gel column chromatography, with low polar solvent elder generation wash-out decon, promptly obtain total diterpene ortho ester compounds with high polar solvent wash-out, wherein, low polar solvent can be chloroform, different ratios sherwood oil-acetone, the different ratios petroleum ether-ethyl acetate, different ratios chloroform-methanol etc.; High polar solvent can be chloroform, different ratios sherwood oil-acetone, different ratios petroleum ether-ethyl acetate, different ratios chloroform-methanol.
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Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102344455A (en) * | 2011-08-13 | 2012-02-08 | 中国科学院昆明植物研究所 | Daphnane diterpene orthoester compound, medicinal composition thereof and preparation method and application thereof |
| CN109970698A (en) * | 2019-04-22 | 2019-07-05 | 广西中医药大学 | Compound and its preparation method and application with antioxidation |
| CN111410679A (en) * | 2020-03-19 | 2020-07-14 | 上海市第七人民医院 | Preparation and application of Daphnane type macrocyclic diterpenoid compound |
| CN113041230A (en) * | 2021-03-23 | 2021-06-29 | 浙江尖峰健康科技有限公司 | Capsule shell of soft capsule and soft capsule prepared from capsule shell |
| CN116120334A (en) * | 2022-11-10 | 2023-05-16 | 沈阳药科大学 | Diterpenoid compounds in lilac daphne flower buds, and preparation method and application thereof |
-
2009
- 2009-09-15 CN CN2009101764488A patent/CN101704826B/en not_active Expired - Fee Related
Cited By (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102344455A (en) * | 2011-08-13 | 2012-02-08 | 中国科学院昆明植物研究所 | Daphnane diterpene orthoester compound, medicinal composition thereof and preparation method and application thereof |
| CN109970698A (en) * | 2019-04-22 | 2019-07-05 | 广西中医药大学 | Compound and its preparation method and application with antioxidation |
| CN109970698B (en) * | 2019-04-22 | 2021-07-27 | 广西中医药大学 | Compound with antioxidant effect and preparation method and application thereof |
| CN111410679A (en) * | 2020-03-19 | 2020-07-14 | 上海市第七人民医院 | Preparation and application of Daphnane type macrocyclic diterpenoid compound |
| CN111410679B (en) * | 2020-03-19 | 2023-05-12 | 上海市第七人民医院(上海中医药大学附属第七人民医院) | Preparation and application of Daphnane type macrocyclic diterpenoid compounds |
| CN113041230A (en) * | 2021-03-23 | 2021-06-29 | 浙江尖峰健康科技有限公司 | Capsule shell of soft capsule and soft capsule prepared from capsule shell |
| CN116120334A (en) * | 2022-11-10 | 2023-05-16 | 沈阳药科大学 | Diterpenoid compounds in lilac daphne flower buds, and preparation method and application thereof |
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| Publication number | Publication date |
|---|---|
| CN101704826B (en) | 2012-01-25 |
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