CN101538258A - New composition of salvia miltiorrhiza and derivative, preparation method and medicine application thereof - Google Patents
New composition of salvia miltiorrhiza and derivative, preparation method and medicine application thereof Download PDFInfo
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Abstract
The invention relates to a new composition of salvia miltiorrhiza and a derivative, a preparation method and application thereof, belonging to the technical field of medicine, wherein the three compositions are salvia miltiorrhiza new lactone A, salvia miltiorrhiza new lactone B and salvia miltiorrhiza new lactone C and derivative thereof and effective parts, which have a (1) chemical structure general formula. The invention also relates to application of a new skeleton composition and a derivative and an effective part thereof on preparing medicine, in particular to the application of same on preparing spiritual class medicine and anticancer class medicine, wherein R1 is selected from hydrogen, alkyl, undersaturated alkyl, hydroxide radical, alkoxyl, carboxyl, esterifiable carboxyl, balmy substituent, halogen, and the like, R2, R3 and R4 are selected from hydrogen, alkyl, undersaturated alkyl, balmy substituent, low grade sugar indican, polysaccharide indican, metallic ions and organic acid ester group, and the R2, the R3 and the R4 can be same and also can be different.
Description
Technical field
The invention belongs to medical technical field, relate to compound new in the red sage root and derivative thereof, its preparation method and medicinal use.
Background technology
Red sage root Salvia miltiorrhiza Bge. is one of widely used medicine in China's traditional medicine.As the medicine of treatment cardiovascular disorder, the medication basis is wide, and evident in efficacy, and clinical application is with a long history.Modern medicine study and clinical practice show that the red sage root has coronary artery dilator, and coronary blood flow increasing, reducing heart rate improve the effect of myocardial anoxia, are usually used in treating coronary heart disease, stenocardia, uncomfortable in chest, cardio palmus.Myocardial infarction, myocarditis can be treated, acute symptom and electrocardiogram(ECG ischemic change can be improved.The red sage root can suppress platelet aggregation, suppresses hematoblastic release reaction, reduce blood viscosity, blood fat reducing, can treat blood high viscosity syndrome.The red sage root can resist erythrocyte aggregation, alleviate atherosclerosis, can suppress fibronectin is transformed into scleroproein, therefore, has the thrombosis of preventing and anticoagulation.Clinical stenocardia, myocardial infarction and the treatment of diseases such as fibrosis, renal failure of being mainly used in.
Along with fast development of social economy, social pressures increase gradually, and some spiritual class diseases such as dysthymia disorders, senile dementia, schizophrenia and obsession etc. have brought serious harm for personal health and family life.The many side effects of medicine that present commercially available some are treated spiritual class disease are bigger, as anxiety, headache, drowsiness, the numbness in hands and feet etc. of having a sleepless night.Yet there are no the red sage root or its composition is used for anticancer and the treatment psychosis.New framework compound of this class and the thymoleptic that the present invention relates to have the part analog structure, to developing novel spiritual class medicine very big inspiration are arranged.
Summary of the invention:
One of the object of the invention provides the new compound and the derivative thereof of a kind of general formula (I), two of purpose of the present invention provides extraction, the preparation method of such new compound and derivative and efficient part, three of purpose of the present invention also relates to the application aspect the preparation medicine of such new compound and derivative and efficient part thereof, particularly treats the application of medicine for senile dementia and anticancer class medicine aspect.
The said class new compound of the present invention (comprising the new lactone A of the red sage root, the new lactone B of the red sage root, the new lactone C of the red sage root) and its general structure of derivative thereof are as follows:
Wherein: R
1Be selected from carboxyl, aromatic substituent, halogen of hydrogen, alkyl, unsaturated alkyl, hydroxyl, alkoxyl group, carboxyl, esterification etc.; R
2, R
3, R
4Be selected from (R such as hydrogen, alkyl, unsaturated alkyl, aromatic substituent, low-grade sugar glucosides, saccharan glucosides, metal ion, organic acid ester group
2, R
3, R
4Can be identical also can be different).
The new lactone A of new compound 1 red sage root described in the present invention, its chemical structure is as follows:
The new lactone B of new compound 2 reds sage root, its chemical structure is as follows:
The new lactone C of new compound 3 reds sage root, its chemical structure is as follows:
The preparation method of its compound is as follows:
Select the commercially available salvia piece of making for the labiate red sage root (Salvia miltiorrhiza Bge.) root and rhizome for use, or the red sage root of fresh collection or the root and rhizome or other positions that belong to other plant together are raw material, water heating with 20 times of amounts decocts extraction 3 times, each two hours, united extraction liquid and concentrating under reduced pressure, X-5 type macroporous resin on the concentrated solution, the difference water, 30% ethanol water, 50% ethanol water, 95% ethanol water elution, concentrating under reduced pressure reclaims solvent, obtain washing part 400g, 30% ethanol washing part 100g, 50% ethanol washing part 50g, 95% ethanol water elution part 18g.
Getting 95% ethanol water elution part 18g is the eluent gradient wash-out through silica gel column chromatography with the chloroform-methanol solvent systems, obtain 18 cuts, wherein cut 12 (0.6g) is the eluent gradient wash-out through silica gel column chromatography with the chloroform-methanol solvent systems once more, obtaining 3 cuts, is that the moving phase separation obtains compound 1 (60mg) with cut 2 (0.2g) with methyl alcohol through Sephadex LH-20.
Getting 50% ethanol water elution part 50g is the eluent gradient wash-out through silica gel column chromatography with the chloroform-methanol solvent systems, obtains 25 cuts, is that moving phase is separated and obtained compound 3 (100mg) with cut 8 (0.7g) with methyl alcohol through Sephadex LH-20.Cut 18 (1.2g) being gone up once more through silica gel column chromatography chloroform-methanol solvent systems gradient elution, obtained 5 cuts, is that the moving phase separation obtains compound 2 (30mg) with cut 3 (0.1) with methyl alcohol through Sephadex LH-20.
The present invention can also be prepared into the derivative that contains the said structure general formula and the medicine of efficient part.
Wherein the derivative of this compounds can be but be not limited to following several:
A. derivative methylates: turn to example with the new lactone A of red sage root methyl, new lactone A is dissolved among the DMSO with the red sage root, adds suitable NaOH, reacts under 80～100 temperature, reaction is finished substantially when treating solution near neutrality, and recrystallization obtains the new lactone A of the red sage root of methyl derivatize.
Turn to example with the new lactone A of red sage root 14-OH methyl, generate the new lactone A of the red sage root derivative that methylates, the example structure formula is:
B. halogen substituted derivative: be substituted by example with the new lactone A of red sage root 14-OH halogen
A. chloro: under argon shield, the new lactone A of the red sage root is dissolved in the dry methylene chloride, add zinc chloride then, add thionyl chloride at last, stir under the certain temperature.With saturated sodium bicarbonate termination reaction, organic layer washing, drying, solvent evaporated.Column chromatography is separated, and obtains the new lactone A of the red sage root of chloro derivatize with sherwood oil-acetone gradient elution.
B. bromo: under argon shield, the new lactone A of the red sage root stirred and add tetrahydrofuran (THF) dissolved NBS, drip tetrahydrofuran (THF) dissolved triphenyl phosphorus afterwards, stirring reaction under the certain temperature, decompression steams solvent, and residue water and ether are washed, and steam solvent.Column chromatography is separated, and obtains the new lactone A of the red sage root of bromo derivatize with sherwood oil-acetone gradient elution.
C. iodo: in container, add the new lactone A of the red sage root, triphenyl phosphorus, imidazoles, iodine and toluene, argon shield is stirred down, refluxes under the certain temperature.After the reaction, remove precipitation, add Na
2S
2O
3The aqueous solution is removed water layer, keeps organic layer.The organic layer washing, anhydrous sodium sulphate dewaters, and decompression steams solvent.Column chromatography is separated, and obtains the new lactone A of the red sage root of iodo derivatize with sherwood oil-acetone gradient elution.
B. glycosides derivative: new lactone A is an example with the red sage root, with monose or oligosaccharides reflux under dry HCl condition such as the disaccharide that forms by above-mentioned monose polymerization, trisaccharide such as the new lactone A of the red sage root and glucose, rhamnosyl, wood sugar, seminoses, use ethyl acetate extraction after reaction is finished, recrystallization promptly gets the new lactone A. of the red sage root of glucosides class derivatize
Be combined into example with monose such as the red sage root new lactone A 14-OH and glucose, rhamnosyl, wood sugar, seminose or oligosaccharides such as the disaccharide that is formed by above-mentioned monose polymerization, trisaccharide, generate glycosides compound, the example structure formula is:
D. metal-salt derivative: the phenolic hydroxyl group among the new lactone A of the red sage root has acidity, can with Na
+, K
+, Mg
2+, Ca
2+, Fe
3+, Cu
2+, Zn
2+Deng alkalimetal ion in conjunction with generating the metal-salt derivative.With the red sage root new lactone A 14-OH and Na
+, K
+, Mg
2+, Ca
2+, Fe
3+, Cu
2+, Zn
2+Is example Deng alkalimetal ion in conjunction with generating the metal-salt derivative, and example derivant structure formula is:
E. lipid derivant: new lactone A is an example with the red sage root; under in argon shield, the new lactone A of the red sage root is dissolved in the dry methylene chloride; add R-C (O)-OH then; DMAP and DCC; stir under the certain temperature; after the reaction end, impurity such as DCU that generates after filtering away unreacted DCC and reacting and DMAP.Steam solvent,, stir, leave standstill and remove by filter impurity, steam solvent with the ether dissolving with the reactant porphyrize.Column chromatography is separated, and obtains the new lactone A of the esterification deutero-red sage root with sherwood oil-acetone gradient elution.
The red sage root new lactone A 14-OH and small molecular organic acids such as formic acid, acetate or diacetyl oxide, butyric acid or butyryl oxide, Succinic Acid or Succinic anhydried, phenylformic acid or benzoyl oxide, acid anhydrides or carboxylic acid halides generate organic acid acetic, and example derivant structure formula is
Macromolecule organic acids such as the red sage root new lactone A 14-OH and palmitinic acid, linolic acid, oleic acid, lauric acid generate organic acid acetic, and example derivant structure formula is:
F. the new lactone A of red sage root 7-OH, 8-OH; The new lactone B of red sage root 7-OH, 8-OH, 14-OH; The new lactone C of red sage root 7-OH, the described derivative reaction of above-mentioned A～E takes place in 8-OH, 14-OH, generates corresponding derivatize product.
G. semi-annular jade pendant base or nitro-derivative: new lactone A is an example with the red sage root, new lactone A of the red sage root and strong sulfuric acid response, (15～25 ℃) generate and face a new lactone A of the sulfonic group derivatize red sage root at a lower temperature, and (80～100 ℃) generate the new lactone A of the contraposition sulfonic group derivatize red sage root under comparatively high temps.The new lactone A of the red sage root and rare nitric acid can generate the new lactone A of the nitro derivatize red sage root at a lower temperature.
Generation is with the red sage root new lactone A 8-OH and sulfuric acid, nitric acid generate the semi-annular jade pendant base or nitro-derivative is an example, and example derivant structure formula is:
H. the new lactone A of red sage root 7-OH, 14-OH; The new lactone B of red sage root 7-OH, 8-OH, 14-OH; The new lactone C of red sage root 7-OH, the described derivative reaction of above-mentioned G takes place in 8-OH, 14-OH, generates corresponding derivatize product.
I. have acidity with the new lactone B of red sage root 17-COOH, can with Na
+, K
+, Mg
2+, Ca
2+, Fe
3+, Cu
2+, Zn
2+In conjunction with generating the metal-salt derivative, is example with the new lactone B17-COOH of the red sage root Deng alkalimetal ion, and example derivant structure formula is:
J. generating lipid derivant with new lactone B17-COOH of the red sage root and alcohols generation esterification is example, adopts the Vesley method, and new lactone B17-COOH of the red sage root and alcohols (R-OH) esterification take place generate the new lactone B of the lipid derivatize red sage root, and yield can reach more than 90%.
Example structure is as follows:
K. reacting with the new lactone B17-COOH of the red sage root and ammonia or amine, to generate amide derivatives be example, under argon shield the new lactone B of the red sage root dissolved in the dry methylene chloride, adds R-NH then
2, DMAP and DCC stir under the certain temperature, wait reaction to finish after, filters away impurity such as the DCU of unreacted DCC and the generation of reaction back and DMAP.Steam solvent,, stir, leave standstill and remove by filter impurity, steam solvent with the ether dissolving with the reactant porphyrize.Column chromatography is separated, and obtains the new lactone B of the amination deutero-red sage root with sherwood oil-acetone gradient elution.
Example structure is as follows, as
L. with the new lactone C of red sage root 17-COOCH
3It is example that ester hydrolysis reaction generation carboxylic acid takes place, and the example structure formula is as follows:
M. the new lactone C of red sage root 17-COOCH
3Under alkaline condition, take place after ester hydrolysis reaction generates carboxylic acid above-mentioned I～K reaction to take place, generate corresponding derivative.
The invention provides a class is new in the red sage root compound and derivative thereof, its preparation method is simple, and the compound of preparation has good anticancer and antidepressant effect.
Description of drawings:
Fig. 1 is the new lactone A's of the red sage root
1H-NMR (MeOH, 300Hz)
Fig. 2 is the new lactone A's of the red sage root
13C-NMR (MeOH, 75Hz)
Fig. 3 is the new lactone A's of the red sage root
13C-NMR (MeOH, 75Hz)
Fig. 4 be the new lactone A of the red sage root HSQC (MeOH, 600Hz)
Fig. 5 be the new lactone A of the red sage root HBCC (MeOH, 600Hz)
Fig. 6 is the new lactone B's of the red sage root
1H-NMR (MeOH, 600Hz)
Fig. 7 is the new lactone B's of the red sage root
13C-NMR (MeOH, 125Hz)
Fig. 8 be the new lactone B of the red sage root DEPT (MeOH, 125Hz)
Fig. 9 is the new lactone C's of the red sage root
1H-NMR (MeOH, 300Hz)
Figure 10 is the new lactone C's of the red sage root
13C-NMR (MeOH, 75Hz)
Figure 11 be the new lactone C of the red sage root HSQC (DMSO, 600Hz)
Figure 12 be the new lactone C of the red sage root HBCC (DMSO, 600Hz)
Figure 13 is the structural formula of compound 1 and derivative thereof
Embodiment:
Embodiment 1: pharmacodynamic experiment:
(1) anti-senile dementia activity research
Anti-microglia pharmacological activation screening.
The chronic inflammatory reaction of microglia activation mediation be in the generation, evolution of nerve degenerative diseases important step, the activation that suppresses microglia may become a new target spot of drug discovery.LPS activates microglia and discharges NO, pro-inflammatory cytokine and active oxygen etc.This experiment is by setting up the screening platform that external LPS activates N9 microglia abnormal activation, and to discharge NO be index to activate microglia, and screening has microglia and activates inhibiting compound in the compound of 3 kinds of extraction separation.
Experiment material:
(1) cell: N9 microglia
(2) reagent: foetal calf serum Fetal bovine serum (Gibco BRL, Grand Island, USA); The IMDM substratum (Gibco BRL, Grand Island, USA); LPS (E5:055) (Sigma, St.Louis, MO, USA); MTT (Sino-AmericanBiotechnology, Beijing, China); Miaow promise ring element (MINO).
(3) given the test agent: the new lactone A of the red sage root, the new lactone B of the red sage root, the new lactone C of the red sage root
Experimental technique:
(1) the mouse microglia is the cultivation of N9
All glasswares that use in cell cultures and the modelling and metallic weapon (culturing bottle, transfer pipet, solution bottle etc.) are all through 121 ℃ of autoclaving 30min, with the LPS of thorough removal pollution.Be mixed with the cell culture fluid that includes 5% calf serum, 100U/mL penicillin and 100U/mL Streptomycin sulphate and 50 μ M 2 mercapto ethanols as the basis with the IMDM substratum.Microglia is with about 4 * 10
5The concentration of cells/mL is at 5%CO
2, the cultivation of going down to posterity in 37 ℃ of culturing bottles accounts for culturing bottle floorage 50-60% to the 3rd day attached cell, with the trysinization attached cell, is passaged to another culturing bottle.As the first-generation, select 3-8 to experimentize with the N9 behind-70 ℃ of Ultralow Temperature Freezer cryopreservation resuscitations for the N9 cell.
(2) medicine collocation method
Compound is configured to the 100mM storing solution with DMSO, keeps in Dark Place in-20 ℃.Facing the time spent is diluted to respective concentration and experimentizes with containing 1% serum I MDM nutrient solution.When the sample of DMSO configuration experimentized, the final concentration of DMSO was less than 1 ‰.
(3) Griess method detection compound activates the restraining effect of microglia to LPS
The N9 microglia of taking the logarithm vegetative period transfers to 5 * 10 with the fresh IMDM substratum that contains 5% foetal calf serum with cell density
5Cells/mL is inoculated in 96 holes pull, 100 μ l/well, and in 37 ℃, 5%CO
2Incubator in cultivate.Change the fresh medium of serum-free behind the cell attachment cultivation 24h into, carry out dosing simultaneously and handle.Sample is established dosage 0.3,3,30 μ M and LPS acting in conjunction.Establish blank and positive control miaow promise ring element (MINO, 20 μ M) simultaneously.The LPS final concentration is 1 μ g/mL in each administration group and the positive controls.After continuing after the cell dosing to cultivate 48h, collect supernatant liquor, the Griess colorimetry detects NO in the supernatant liquor
2-Content.
(4) the mtt assay detection compound is to the influence of microglia cell survival rate
The N9 microglia of taking the logarithm and cultivating vegetative period transfers to 5 * 10 with the fresh IMDM substratum that contains 5% foetal calf serum with cell density
5Cells/mL is inoculated in 96 holes pull, 100 μ l/well, and in 37 ℃, 5%CO
2Incubator in cultivate.Cell attachment changes fresh medium into after cultivating 24h, carries out dosing simultaneously and handles.Sample is established dosage 0.3,3,30 μ M and LPS acting in conjunction.Establish blank and positive control simultaneously.The LPS final concentration is 1 μ g/mL in each administration group and the positive controls.Continue to cultivate 48h after the cell dosing, add MTT solution then in enchylema, 10 μ l/well are hatched 3h jointly with cell and 0.25mg/mLMTT under 37 ℃, absorb nutrient solution, add isopyknic DMSO solution then, measure its optical density(OD) OD value.Data processing utilizes the microplate reader corresponding software to carry out data processing, calculates the mean value of each three holes of sample OD value, utilize mean value be calculated as follows cell survival rate (cell viability, CV%).
Mean value * 100% of the mean value of cell survival rate %=sample sets OD value/blank group OD value
CV％＝ODsample/ODcontrol×100％
The statistical procedures method
All data adopt the analysis of testing of SPSS (11.5) statistical packages.The result represents that with mean value ± standard error mean relatively carries out the homoscedasticity analysis between group, and carries out Dunnett ' s test analytical procedure and organize a comparison and Student ' s test statistical procedures.
Experimental result:
Three kinds of samples of table 1 activate the influence (M+SE) that the N9 microglia discharges nitrogen protoxide (%) to LPS
###: compare P＜0.001 with the blank group;
*Compare P＜0.05 with model control group;
*: compare P＜0.01 with model control group;
* *, compare P＜0.001 with model control group.
Three kinds of compounds of table 2 are to N9 microglia survival rate (%) influence (M+SE)
Experiment conclusion:
(1) the new lactone A of the medicine red sage root, the new lactone B of the red sage root, the new lactone C of the red sage root can discharge NO by inhibition activated microglia in various degree, see Table 1.
(2) wherein the new lactone A of the red sage root, the new lactone C of red sage root specific activity positive drug are strong, and NO is discharged inhibiting IC
50＜20 μ M.
(2) Anticancer Activities
Suppress the screening of HL-60 cell strain proliferation activity
At ginseng new lactone A, the new lactone B of the red sage root, the new lactone C of red sage root human leukemia cell line HL-60, adopt classical mtt assay to carry out antitumor activity screening.
Experiment material
(1) is subjected to the reagent product: join new lactone A, the new lactone B of the red sage root, the new lactone C of the red sage root
(2) experimental cell strain and source: HL-60: people's acute myeloid leukaemia cell (available from ATCC)
(3) experiment reagent: RMPI RPMI-1640 (available from Gibco), foetal calf serum (TBD company), tetramethyl-azo azoles salt (MTT) (U.S. Sigma), dimethyl sulfoxide (DMSO) (DMSO), NaCl, KCl, KH
2PO3, Na
2HPO3, NaHCO3, microplate reader (Austrian TECAN), 96 porocyte culture plates (Costar company)
Experimental technique
1, drug treating
(1) dissolving of medicine
Compound is joined new lactone A, the new lactone B of the red sage root, the new lactone C of the red sage root and is Powdered, uses the DMSO dissolving.Be made into the mother liquor that concentration is 100mmol/L, be stored in-20 ℃.Facing the time spent is 100 μ mol/L with corresponding nutrient solution with its dilution, 10 μ mol/L, and 1 μ mol/L experimentizes.When the sample of DMSO configuration experimentized, the final concentration of DMSO was 1 ‰.Choose YH-21 (Yuanhuacine) as positive control drug, concentration is 10 μ mol/L.
(2) administration is handled
The cell of taking the logarithm vegetative period is adjusted suitable cell density, is inoculated in 96 orifice plates, and 100 μ L/well are incubated at 37 ℃, 5%CO
2Incubator in.After cultivating 24h, be 100 μ mol/L with drug dilution, 10 μ mol/L, three concentration of 1 μ mol/L, 10 μ L/well, effect 48h.Set up blank group, administration group separately, establish 5 multiple holes for every group.
2, MTT check
(1) ultimate principle of mtt assay
Cell survival rate is measured and is adopted the MTT analytical method, and ((4,5-dimethyl-2thiahiazoyl)-3,5-di-phenyl-tetrazoliumbromide MTT) is the basis to 3-with viable cell metabolism reduction tetramethyl-azo azoles salt.MTT is a yellow compound, it is the hydrionic dyestuff of a kind of acceptance, can act on the respiratory chain in the viable cell plastosome, tetrazole ring opening under the effect of succinodehydrogenase and cytochrome C, generate blue Formazan crystallization, Formazan crystalline growing amount only is directly proportional with the viable cell number.This enzyme disappears in the dead cell, MTT can not be reduced.The Formazan crystallization that is reduced into generation can contain 50%, N, dissolve in the MTT lysate of N-dimethylformamide and 20% sodium laurylsulfonate (PH4.7), utilize microplate reader to measure the optical density(OD) OD value that 490nm goes out, the size of OD value is directly proportional with the Formazan crystalline amount that is generated, thereby reflects the influence of medicine pair cell surviving rate.
(2) measuring method of mtt assay
Behind the drug effect 48h, cell and 0.25mg/ml MTT are hatched 4h jointly under 37 ℃, every hole adds 100 μ l dimethyl sulfoxide (DMSO) (DMSO) behind the absorption nutrient solution, and the dissolving back uses microplate reader to measure its optical density(OD) OD value in 490nm fully.Be 100% with blank group OD value at last, calculate and respectively organize cell inhibitory rate.
3, statistical method
All data adopt the analysis of testing of SPSS (13.0) statistical packages.Each organize data with the mean value standard error (Mean ± S.E.) expression adopts One-Way ANOVA to estimate globality difference, and carry out Dunnett or Dunnett ' s T3 check organize between relatively.
4, the method for calculation of IC50
Parameters such as each dosage and inhibiting rate are calculated IC50 with nonlinear regression and fitting.
Experimental result
The inhibiting rate that table 1 compound survives tumour cell HL-60 (%) (M ± SE)
***P＜0.001 Vs Control；
**P＜0.01 Vs Control；
*P＜0.05Vs Control
The inhibiting rate (%) that table 2 compound YH-21 survives tumour cell HL-60 (M ± SE)
| control | 6.25μmol/L | 12.5μmol/L | 25μmol/L | 50μmol/L | IC50μmol/L | |
| YH-21 | 0.00±0.18 | 24.041±0.24 *** | 41.20±0.08 *** | 82.96±0.07 *** | 86.653±0.15 *** | 12.90 |
Experimental result
(1) can find out that by above-mentioned experimental result after the drug effect 48h, the HL-60 cell strain all has certain lethal effect in three kinds of compounds.
(2) wherein the new lactone A of red sage root specific activity positive drug is strong.
Embodiment 2:
Structure elucidation:
Compound (1) yellow powder (methyl alcohol), [α]
D 2020.4 (c=0.260, MEOH), high resolution mass spectrum provides quasi-molecular ion peak m/z 319.0575[M+Na]
+(calculated value is 319.0577) determines that molecular formula is C
17H
12O
5IR (KBr) spectrum 3425cm
-1Signify hydroxy absorbs, 1746cm
-1Show that lactone absorbs, 1602,1571cm
-1Show that phenyl ring absorbs.(MeOH 300Hz) and in the HMQC spectrum provides a methyl hydrogen signal δ: 0.98 (3H, d, 7.2, δ according to 1H-NMR
C22.8); The methyne hydrogen signal δ of a SP3 hydridization: 4.96 (1H, q, 7.2, δ
c35.9); Two adjacent hydrogen signal δ on two groups of phenyl ring
H: 6.67 (d, 1H, J=8.4, δ
c113.6) and δ
H: 7.06 (d, 1H, J=8.4, δ
c129.4), δ
H6.79 (d, 1H, J=8.4, δ
c124.4) and δ
H6.75 (d, 1H, J=8.4, δ
c120.2); And the hydrogen signal δ on two key
H7.66 (s, 1H, δ
c141.4).
13(MeOH provides 17 carbon signals in 75Hz) to C-NMR, two saturated carbon signals wherein, the carbon signal of 14 two keys, a carbonyl carbon signal.Binding molecule formula this compound as can be known has 12 degrees of unsaturation, removes seven pairs of two keys and a carbonyl, and also surplus 4 degrees of unsaturation point out this compound to be made up of four rings.
H-10 and C-4 in the relevant spectrum of HMBC, C-8, C-9, C-11, C-16 has relevant with C-17; H-12 and C-10, C-11, C-14 has relevant with C-16; H-5/C-3 has relevant with C-9; H-3 and C-2, C-5, C-9 has relevant this compound of prompting that the triatomic ring structure of individual dibenzocycloheptane is arranged with C-16.H-3 and C-1 in the relevant spectrum of HMBC, C-2 has relevant with C-16; H-13 and C-11, C-14, there is relevant this compound that not only illustrates to have a lactone ring five membered with C-15, and be connected with a hydroxyl on the explanation C-14 position. only have relevant carbonyl as can be seen directly to link to each other with C-2 with H-3 and formed a lactonic ring with C-15 from the carbonyl carbon signal, this also is made up of four ring structures with the top compound of inferring and conforms to.According to
13H-10 and C-4 in the relevant spectrum of chemical displacement value in the C-NMR spectrum and HMBC, C-8 has relevant with C-9; H-6 and C-4, C-7 has relevantly with C-8, illustrates that C-7 and C-8 position are connected with two hydroxyls respectively.According to H
3-17 and C-9, C-10 has relevant methyl as can be seen to link to each other with C-10 with C-11.Like this compound once two dimensional structure be determined.The hydrocarbon ownership of compound one and HMBC are correlated with and see Table 1.
Compound (2) yellow powder (methyl alcohol), [α]
D 2015.0 (c=0.200, MEOH), high resolution mass spectrum provides quasi-molecular ion peak m/z 363.0476[M+Na]
+(calculated value is 363.0475) determines that molecular formula is C
18H
12O
7IR (KBr) spectrum 3437cm
-1Signify hydroxy absorbs, 1743cm
-1Show that lactone absorbs, 2500,1740cm
-1Show that carboxyl absorbs, 1600,1565cm
-1Show that phenyl ring absorbs.According to
1H-NMR (MeOH, 600Hz) and
13C-NMR (MeOH, 125Hz) this compound has identical constitutional features with compound 1 as can be seen, and just the methyl of high field region has become methylene radical in the 1H-NMR spectrum, and
13In the C-NMR spectrum in low place a carboxyl carbon signal has appearred.From on can release the methyl of compound 1 a hydrogen proton and replaced by carboxyl and just become compound 2, this is C with compound 2 molecular formula also
18H
12O
7Conform to.The hydrocarbon ownership of compound 2 is seen Table 1.
Compound (3) yellow powder (methyl alcohol), [α]
D 2016.3 (c=0.245, MEOH), high resolution mass spectrum provides quasi-molecular ion peak m/z 377.0638[M+Na]
+(calculated value is 377.0632) determines that molecular formula is C
19H
14O
7IR (KBr) spectrum 3409cm
-1Signify hydroxy absorbs, 1764cm
-1Show that lactone absorbs 1678cm
-1Show that ester carbonyl group absorbs, 1600,1572cm
-1Show that phenyl ring absorbs.According to 1H-NMR (MeOH, 300Hz) and
13C-NMR (MeOH, 75Hz) this compound has identical constitutional features with compound 2 as can be seen, and just a methoxyl group signal has appearred in high field region in the 1H-NMR spectrum, and
13The carbon signal that has occurred a methoxyl group in the C-NMR spectrum at high field region.Can see that according to the relevant spectrum of HMBC the hydrogen signal on the methoxyl group only has relevantly with the carbonyl carbon signal, illustrate that this methoxyl group becomes ester just to become compound 3 with carboxyl in the compound 2, this also with compound 3 molecular formula be C
19H
14O
7Conform to.The hydrocarbon ownership of compound 2 is seen Table 1.
Table 1.The
1H and
13C NMR Data for compound 1，2，and 3
Claims (6)
1. new compound and derivative thereof in the red sage root, it is characterized in that: described compound and derivative thereof have following general structure:
Wherein: R
1Be selected from carboxyl, aromatic substituent, halogen of hydrogen, alkyl, unsaturated alkyl, hydroxyl, alkoxyl group, carboxyl, esterification etc.; R
2, R
3, R
4Be selected from hydrogen, alkyl, unsaturated alkyl, aromatic substituent, low-grade sugar glucosides, saccharan glucosides, metal ion, organic acid ester group, R
2, R
3, R
4Can be identical also can be different.
2. new compound and derivative thereof in the red sage root according to claim 1, it is characterized in that: described compound and structural formula thereof are:
The new lactone C of the new lactone B of the new lactone A of the red sage root red sage root red sage root
3. new compound and derivative thereof can be used as the pharmaceutical composition that activeconstituents and one or more pharmaceutically acceptable carriers are formed in the claim 1 or the 2 described reds sage root.
4. the new compound and the preparation method of derivative thereof in the red sage root as claimed in claim 1 is characterized in that the preparation method of described this compound is as follows:
Select the commercially available salvia piece of making for the labiate red sage root (Salvia miltiorrhiza Bge.) root and rhizome for use, or the red sage root of fresh collection or the root and rhizome or other positions that belong to other plant together are raw material, water heating with 20 times of amounts decocts extraction 3 times, each two hours, united extraction liquid and concentrating under reduced pressure, X-5 type macroporous resin on the concentrated solution, the difference water, 30% ethanol water, 50% ethanol water, 95% ethanol water elution, concentrating under reduced pressure reclaims solvent, obtain washing part 400g, 30% ethanol washing part 100g, 50% ethanol washing part 50g, 95% ethanol water elution part 18g;
95% ethanol water elution part 18g is the eluent gradient wash-out through silica gel column chromatography with the chloroform-methanol solvent systems, obtain 18 cuts, wherein 0.6g cut 12 is eluent gradient (100: 0 to 0: 100) wash-out through silica gel column chromatography with the chloroform-methanol solvent systems once more, obtaining 3 cuts, is that the moving phase separation obtains 60mg compound 1 with 0.2g cut 2 with methyl alcohol through Sephadex LH-20;
Getting 50% ethanol water elution part 50g is eluent gradient (100: 0 to 0: 100) wash-out through silica gel column chromatography with the chloroform-methanol solvent systems, obtaining 25 cuts, is that the moving phase separation obtains 100mg compound 3 with 0.7g cut 8 with methyl alcohol through Sephadex LH-20.1.2g cut 18 being gone up once more through silica gel column chromatography chloroform-methanol solvent systems gradient elution, obtained 5 cuts, is that the moving phase separation obtains 30mg compound 2 with 0.1g cut 3 with methyl alcohol through Sephadex LH-20.
5. new compound and the application of derivative in the spiritual class disease medicament of preparation treatment thereof in the claim 1 or 2 the described red sage root.
6. new compound and the application of derivative in the anticancer class medicine of preparation thereof in the claim 1 or 2 the described red sage root.
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Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102670623A (en) * | 2012-06-12 | 2012-09-19 | 昆明理工大学 | Application of tanshinlactone in preparation of anti-tumor-angiogenesis medicament |
| CN114591277A (en) * | 2022-02-28 | 2022-06-07 | 南京林业大学 | 5/6/7/6 tetracyclic compound fused with benzene ring and synthesis method thereof |
| WO2023284262A1 (en) * | 2021-07-13 | 2023-01-19 | 青岛农业大学 | 3,4-fused seven-membered heterocyclic oxindole, synthesis method therefor and application thereof |
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| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1315495C (en) * | 2005-02-04 | 2007-05-16 | 邢为藩 | Danshen root extract, production, medicine preparation and inspection thereof |
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2009
- 2009-03-04 CN CN 200910010555 patent/CN101538258B/en not_active Expired - Fee Related
Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102670623A (en) * | 2012-06-12 | 2012-09-19 | 昆明理工大学 | Application of tanshinlactone in preparation of anti-tumor-angiogenesis medicament |
| WO2023284262A1 (en) * | 2021-07-13 | 2023-01-19 | 青岛农业大学 | 3,4-fused seven-membered heterocyclic oxindole, synthesis method therefor and application thereof |
| CN114591277A (en) * | 2022-02-28 | 2022-06-07 | 南京林业大学 | 5/6/7/6 tetracyclic compound fused with benzene ring and synthesis method thereof |
| CN114591277B (en) * | 2022-02-28 | 2022-11-25 | 南京林业大学 | 5/6/7/6 tetracyclic compound fused with benzene ring and synthetic method thereof |
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