CN111875513A - Resveratrol A ring N (CH)3)2Base derivatives, preparation method and application thereof - Google Patents
Resveratrol A ring N (CH)3)2Base derivatives, preparation method and application thereof Download PDFInfo
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Abstract
The invention provides resveratrol A-ring N (CH)3)2A derivative, a preparation method and application thereof, belonging to the technical field of medicine. The invention provides resveratrol A ring N (CH) with a structure shown in a formula I3)2The derivative, as a sodium hydrogen exchanger-1 (NHE1) receptor antagonist, acts on an NHE1 receptor in a targeting way, and simultaneously inhibits 2 signal conduction paths of a phosphatidylinositol 3-kinase/protein serine threonine kinase (PI3K/AKT) signal path and a Janus kinase/signal transduction and transcription activator (JAK/STAT), thereby playing a role in treating alcoholic fatty liver.
Description
Technical Field
The invention relates to the technical field of medicines, in particular to resveratrol A-ring N (CH)3)2A derivative, a preparation method and application thereof.
Background
Alcoholic fatty liver is a liver disease caused by long-term drinking, and is one of the types of alcoholic liver diseases. Alcoholic fatty liver is a common pathological change of the liver, not an independent disease. When liver cells are damaged, alanine Aminotransferase (ALT) and aspartate Aminotransferase (AST) overflow causes the ALT and AST contents in serum to be increased, so the degree of damage of the liver cells of the alcoholic fatty liver patients and the repair condition of the liver cells can be reflected by measuring the ALT and AST activities in the serum.
At present, the clinical treatment of alcoholic fatty liver does not have a unified standard, and commonly used auxiliary intervention means comprise insulin sensitizers, lipid regulating drugs, antioxidants and the like. Common drugs for treating alcoholic fatty liver include: western medicines such as choline methionine, lecithin, silymarin, inosine, coenzyme A, carnitine orotate and the like mainly have the functions of protecting liver cells and increasing fat transportation; ② antioxidants such as resveratrol, reduced glutathione, taurine, vitamin E, mainly for inhibiting cholesterol, triglyceride oxidation and lipid accumulation; ③ the Chinese herbs such as curcuma longa, prepared fleece-flower root and hawthorn fruit, etc., mainly have the functions of reducing blood fat and preventing cholesterol from depositing in the liver.
In the prior art, resveratrol has a good antioxidation effect. However, resveratrol is not ideal in its therapeutic effect on alcoholic fatty liver.
Disclosure of Invention
In view of the above, the present invention aims to provide a resveratrol A ring N (CH)3)2A derivative, a preparation method and application thereof. The invention providesResveratrol A Ring N (CH) supply3)2The derivative has therapeutic effect on alcoholic fatty liver.
In order to achieve the above object, the present invention provides the following technical solutions:
the invention provides resveratrol A-ring N (CH)3)2A group derivative having the structure shown in formula I:
the invention also provides resveratrol A ring N (CH) in the technical scheme3)2A process for the preparation of a derivative comprising the steps of:
mixing diethyl phosphite, an organic solvent, NaH and 4-methoxybenzyl bromide to carry out debromination reaction to obtain a compound with a structure shown in a formula 1;
mixing 4-dimethylamino salicylaldehyde, an organic solvent, chloromethyl methyl ether and diisopropylethylamine for a substitution reaction to obtain a compound with a structure shown in a formula 2;
mixing a compound with a structure shown in a formula 1, a compound with a structure shown in a formula 2, an organic solvent and NaH for a condensation reaction to obtain a compound with a structure shown in a formula 3;
mixing the compound with the structure shown in the formula 3, tert-butyl alcohol and pyridinium p-toluenesulfonate, and refluxing to obtain resveratrol A ring N (CH) with the structure shown in the formula I3)2A derivative of phenyl;
preferably, the time of the debromination reaction is 1-3 h.
Preferably, the time of the substitution reaction is 10-30 min.
Preferably, the condensation reaction is carried out at the temperature of 0-100 ℃ for 1-3 h.
Preferably, the reflux temperature is 90-100 ℃, and the reflux time is 1-3 h.
Preferably, the molar ratio of the compound with the structure shown in the formula 3 to the pyridinium p-toluenesulfonate is 1: 2-2.1.
The invention also provides resveratrol A ring N (CH) in the technical scheme3)2The resveratrol A ring N (CH) prepared by the preparation method of the base derivative or the technical scheme3)2The application of the derivative in preparing medicines for treating alcoholic fatty liver is provided.
Preferably, the drug for treating alcoholic fatty liver disease comprises an effective dose of resveratrol A ring N (CH) with a structure shown in formula I3)2The derivative, the stereoisomer and the pharmaceutically acceptable salt thereof and pharmaceutically acceptable carriers, auxiliary materials, excipients and diluents.
Preferably, the dosage form of the drug for treating alcoholic fatty liver disease comprises pharmaceutically acceptable dosage forms of tablets, injections, capsules, granules, pills, powders, oral liquids, sustained release preparations, controlled release preparations or nano preparations.
The invention provides resveratrol A ring N (CH) with a structure shown in a formula I3)2The derivative, as a sodium hydrogen exchanger-1 (NHE1) receptor antagonist, acts on an NHE1 receptor in a targeting way, and simultaneously inhibits 2 signal conduction paths of a phosphatidylinositol 3-kinase/protein serine threonine kinase (PI3K/AKT) signal path and a Janus kinase/signal transduction and transcription activator (JAK/STAT), thereby playing a role in treating alcoholic fatty liver.
The invention also provides resveratrol A ring N (CH) in the technical scheme3)2The preparation method of the derivative has the advantages of mild conditions, low raw material cost, easy operation and high yield.
Drawings
FIG. 1 shows resveratrol A ring N (CH) prepared by the present invention3)2The signal path diagram of the mechanism of treating alcoholic fatty liver by the derivative is shown;
FIG. 2 shows that resveratrol A ring N (CH) prepared by the present invention3)2Reaction scheme of the derivatives;
FIG. 3 shows the effect of different experimental groups on the initial body mass;
FIG. 4 is a graph of the effect of different experimental groups on end body mass;
FIG. 5 is the effect of different experimental groups on hepatic index;
FIG. 6 is a graph of the effect of different experimental groups on serum TG content;
FIG. 7 is a graph of the effect of different experimental groups on serum TC levels;
FIG. 8 is a graph of the effect of different experimental groups on serum HDL-C levels;
FIG. 9 is a graph of the effect of different experimental groups on serum ALT levels;
FIG. 10 is a graph of the effect of different experimental groups on serum AST levels;
FIG. 11 shows the effect of different experimental groups on serum SOD levels;
FIG. 12 is a graph of the effect of different experimental groups on serum MDA content;
FIG. 13 shows the effect of different experimental groups on the subregional structure of the liver (400X);
FIG. 14 is a graph of the effect of different experimental groups on immunohistochemistry for hepatic NHE-1 expression (400X);
FIG. 15 is a graph of the effect of different experimental groups on hepatic NHE-1 expression of WesternBlot (400X);
FIG. 16 is a statistical plot of the effect of different experimental groups on hepatic NHE-1 expression of WesternBlot;
FIG. 17 shows the effect of different experimental groups on hepatic PI3K and STAT immunofluorescence (400 ×).
Detailed Description
The invention provides resveratrol A-ring N (CH)3)2A group derivative having the structure shown in formula I:
in the invention, the resveratrol A ring N (CH) with the structure shown in formula I3)2Chemical name of the derivative is 2-dihydroxy-4' -methoxy-4-dimethylamino-1, 2-diphenylethylene ((E) -5- (dimethyllami)no) -2- (4-methoxystyryl) phenol) with molecular formula C17H19NO2The molecular weight is 269.
FIG. 1 shows resveratrol A ring N (CH)3)2Resveratrol A-Ring N (CH) as a signaling pathway map for the treatment of fatty liver3)2The derivative, as a sodium hydrogen exchanger-1 (NHE1) receptor antagonist, acts on an NHE1 receptor in a targeting way, and simultaneously inhibits 2 signal conduction paths of a phosphatidylinositol 3-kinase/protein serine threonine kinase (PI3K/AKT) signal path and a Janus kinase/signal transduction and transcription activator (JAK/STAT), thereby playing a role in treating alcoholic fatty liver.
The invention also provides resveratrol A ring N (CH) in the technical scheme3)2A process for the preparation of a derivative comprising the steps of:
mixing diethyl phosphite, an organic solvent, NaH and 4-methoxybenzyl bromide to carry out debromination reaction to obtain a compound with a structure shown in a formula 1;
mixing 4-dimethylamino salicylaldehyde, an organic solvent, chloromethyl methyl ether and diisopropylethylamine for a substitution reaction to obtain a compound with a structure shown in a formula 2;
mixing a compound with a structure shown in a formula 1, a compound with a structure shown in a formula 2, an organic solvent and NaH for a condensation reaction to obtain a compound with a structure shown in a formula 3;
mixing the compound with the structure shown in the formula 3, tert-butyl alcohol and p-toluenesulfonate, and refluxing to obtain resveratrol A ring N (CH) with the structure shown in the formula I3)2A derivative of phenyl;
in the present invention, the raw materials used in the present invention are all commercially available products unless otherwise specified.
In the present invention, the reaction principle of the preparation method is shown in fig. 2.
The invention mixes diethyl phosphite, organic solvent, NaH and 4-methoxybenzyl bromide to carry out debromination reaction, and obtains a compound with a structure shown in a formula 1. In the present invention, the organic solvent is preferably DMF.
In the present invention, the amount ratio of diethyl phosphite, organic solvent, NaH and 4-methoxybenzyl bromide is preferably 10 mmol: 8mL of: 15 mmol: 15 mmol.
In the invention, the time of the debromination reaction is preferably 1-3 h, and the temperature of the debromination reaction is preferably room temperature, so that additional heating or cooling is not required.
According to the invention, preferably, after diethyl phosphite is dissolved in DMF, NaH is added under the condition of ice-water bath, and 4-methoxybenzyl bromide is added after half an hour.
After the debromination reaction is finished, a saturated ammonium chloride solution is preferably added to quench the reaction, then dichloromethane is used for extraction, and column chromatography purification is carried out after reduced pressure distillation to obtain the compound with the structure shown in formula 1. The present invention is not particularly limited to the specific operations of the extraction and vacuum distillation of dichloromethane, and may be performed in a manner well known to those skilled in the art. In the invention, the eluent used for the column chromatography purification is preferably a petroleum ether-ethyl acetate mixed solution, and the volume ratio of petroleum ether to ethyl acetate in the mixed solution is preferably 1: 1.
The method comprises the step of mixing 4-dimethylamino salicylaldehyde, an organic solvent, chloromethyl methyl ether and diisopropylethylamine for substitution reaction to obtain a compound with a structure shown in a formula 2. In the present invention, the organic solvent is preferably dichloromethane.
In the invention, the dosage ratio of the 4-dimethylamino salicylaldehyde, the organic solvent, the chloromethyl methyl ether and the diisopropylethylamine is preferably 10 mmol: 40mL of: 15 mmol: 15 mmol.
In the present invention, the time of the substitution reaction is preferably 10 to 30 min.
In the invention, 4-dimethylamino salicylaldehyde is preferably dissolved in dichloromethane, and chloromethyl methyl ether and diisopropylethylamine are added under the condition of ice-water bath.
After the substitution reaction is finished, preferably, saturated ammonium chloride solution is added to quench the reaction, then dichloromethane is used for extraction, and column chromatography purification is carried out after reduced pressure distillation to obtain the compound with the structure shown in the formula 2. The present invention is not particularly limited to the specific operations of the extraction and vacuum distillation of dichloromethane, and may be performed in a manner well known to those skilled in the art. In the invention, the eluent used for the column chromatography purification is preferably a petroleum ether-ethyl acetate mixed solution, and the volume ratio of petroleum ether to ethyl acetate in the mixed solution is preferably 5: 1.
After obtaining the compounds with the structures shown in the formulas 1 and 2, the invention mixes the compound with the structure shown in the formula 1, the compound with the structure shown in the formula 2, an organic solvent and NaH for condensation reaction to obtain the compound with the structure shown in the formula 3. In the present invention, the organic solvent is preferably DMF.
In the present invention, the compound having the structure represented by formula 1, the compound having the structure represented by formula 2, the organic solvent and NaH are preferably used in a ratio of 3 mmol: 4.5 mmol: 4mL of: 4.5 mmol.
In the invention, the condensation reaction is preferably carried out at the temperature of 0-100 ℃ for 1-3 h. In the present invention, the condensation reaction is preferably carried out in an oil bath.
According to the invention, the compound with the structure shown in the formula 1 is preferably dissolved in DMF, NaH is added under the condition of ice-water bath, and then the compound with the structure shown in the formula 2 is added.
After the condensation reaction is finished, the condensation product is preferably naturally cooled to room temperature, then a saturated ammonium chloride solution is added for quenching reaction, then dichloromethane is used for extraction, and column chromatography purification is carried out after reduced pressure distillation, so as to obtain the compound with the structure shown in the formula 3. The present invention is not particularly limited to the specific operations of the extraction and vacuum distillation of dichloromethane, and may be performed in a manner well known to those skilled in the art. In the invention, the eluent used for the column chromatography purification is preferably a petroleum ether-ethyl acetate mixed solution, and the volume ratio of petroleum ether to ethyl acetate in the mixed solution is preferably 10: 1.
After obtaining the compound with the structure shown in the formula 3, the invention uses the compound with the structure shown in the formula 3, tertiary butanol and p-toluene sulfonic acidMixing with pyridinium sulfate, and refluxing to obtain resveratrol A ring N (CH) with structure shown in formula I3)2And (b) a derivative thereof.
In the invention, the reflux temperature is preferably 90-100 ℃, more preferably 95 ℃, and the time is preferably 1-3 h. In the present invention, the reflux is preferably carried out in an oil bath pan.
In the invention, the molar ratio of the compound having the structure shown in the formula 3 to the pyridinium p-toluenesulfonate is preferably 1:2 to 2.1.
In the present invention, the amount ratio of the compound having the structure represented by formula 3 to t-butanol is preferably 1 mmol: 4 mL.
According to the invention, the compound with the structure shown in the formula 3 is preferably dissolved in tert-butyl alcohol, and then p-toluenesulfonic acid pyridinium salt is added.
After the reflux is finished, the reflux product is preferably naturally cooled to room temperature, water is added for quenching reaction, then dichloromethane is used for extraction, and after reduced pressure distillation and column chromatography purification, the compound with the structure shown in the formula 3 is obtained. The present invention is not particularly limited to the specific operations of the extraction and vacuum distillation of dichloromethane, and may be performed in a manner well known to those skilled in the art. In the invention, the eluent used for the column chromatography purification is preferably a petroleum ether-ethyl acetate mixed solution, and the volume ratio of petroleum ether to ethyl acetate in the mixed solution is preferably 3: 1.
The invention also provides resveratrol A ring N (CH) in the technical scheme3)2The resveratrol A ring N (CH) prepared by the preparation method of the base derivative or the technical scheme3)2The application of the derivative in preparing medicines for treating alcoholic fatty liver is provided.
In the invention, the drug for treating alcoholic fatty liver disease preferably comprises resveratrol A ring N (CH) with the structure shown in formula I, wherein the effective dose is more than or equal to 10mg/kg3)2The derivative, the stereoisomer and the pharmaceutically acceptable salt thereof and pharmaceutically acceptable carriers, auxiliary materials, excipients and diluents. In the present invention, the effective dose is preferably not less than 10 mg/kg.
In the invention, the dosage form of the drug for treating alcoholic fatty liver preferably comprises a pharmaceutically acceptable dosage form of tablets, injections, capsules, granules, pills, powder, oral liquid, sustained release preparations, controlled release preparations or nano preparations.
To further illustrate the present invention, the following examples are given to provide resveratrol A ring N (CH)3)2The derivatives, the preparation and use thereof, and the microbial fuel cell on the sea bottom will be described in detail, but they should not be construed as limiting the scope of the present invention.
Example 1
1. Medicine preparation: resveratrol (Nanjing spring and autumn bioengineering, Inc.); p-methoxybenzyl bromide, 2-hydroxy-5-dimethylaminobenzaldehyde, chloromethyl methyl ether, tetrahydrofuran, sodium hydride, methylene chloride, diethyl phosphite, N-dimethylformamide, pyridinium p-toluenesulfonate, (ann naiji chemical limited); compound methionine choline tablet (Tonghua Dongbao pharmaceutical industry Co., Ltd., China); NHE1 primary antibody (BM5689), PI3K primary antibody (BM4344), STAT primary antibody (M00337) (Strobile bioengineering, GmbH, Wuhan).
Resveratrol A Ring N (CH)3)2The synthesis method of the derivative comprises the following steps:
(1) diethyl phosphite (10mmol, 1.38g) was dissolved in DMF (8mL), NaH (15mmol, 0.36g) was added under ice water bath conditions, and after half an hour 4-methoxybenzyl bromide (15mmol, 3.02g) was added and reacted for 1 hour. After the reaction is completed, adding a saturated ammonium chloride solution to quench the reaction, extracting the reaction by using dichloromethane, carrying out reduced pressure distillation and then carrying out column chromatography purification (petroleum ether: ethyl acetate: 1) to obtain a compound with a structure shown in a formula 1, wherein the yield is 90%;
(2) 4-Dimethylaminosalicylaldehyde (10mmol, 1.65g) was dissolved in dichloromethane (40mL), and chloromethyl methyl ether (15mmol, 1.21g) and diisopropylethylamine (15mmol, 1.94g) were added under ice-water bath conditions for 2 hours. Adding saturated ammonium chloride solution to quench reaction, extracting with dichloromethane, distilling under reduced pressure, and purifying by column chromatography (petroleum ether: ethyl acetate: 5:1) to obtain compound with structure shown in formula 2, wherein the conversion rate is 98%;
(3) the compound having the structure shown in formula 1 (3mmol, 0.774g) was dissolved in DMF (4mL), NaH (4.5mmol, 0.11g) was added under ice-water bath conditions, and after reaction for half an hour, the compound having the structure shown in formula 2 (4.5mmol, 0.94g) was added and placed in an oil bath at 100 ℃ for reaction for 1 hour. After the reaction is completed, cooling to room temperature, adding a saturated ammonium chloride solution to quench the reaction, extracting with dichloromethane, carrying out reduced pressure distillation, and then carrying out column chromatography purification (petroleum ether: ethyl acetate: 10:1) to obtain a compound with a structure shown in a formula 3, wherein the yield is 95%;
(4) a compound having a structure represented by formula 3 (1mmol, 0.313g) was dissolved in t-butanol (4mL), and pyridinium p-toluenesulfonate (2mmol, 0.502g) was added and placed in a 95 ℃ oil bath and heated under reflux for 1 hour. Cooling to room temperature after the reaction is completed, adding water to quench the reaction, extracting with dichloromethane, distilling under reduced pressure, and purifying by column chromatography (petroleum ether: ethyl acetate: 3:1) to obtain resveratrol A ring N (CH) with structure shown in formula I3)2Derivative, yield 84%;
for the prepared resveratrol A ring N (CH)3)2Carrying out structural characterization on the derivative, white solid;1H NMR(400MHz,DMSO-d6):2.88(s,6H),3.75(s,3H),6.23-6.28(m,2H), 6.88-6.92(m,3H),7.15(d,J＝16.4Hz,1H),7.36(d,J＝8.4Hz,1H),7.40(d,J＝8.4Hz,2H),9.41(brs,1H).13C NMR(150MHz,DMSO-d6):40.6,55.5,105.2, 114.6,122.3,123.4,127.2,127.4,127.9,130.6,131.7,151.0,156.1,158.5.
2. animals: Sprague-Dawley (SD) rats, 6 weeks old, male, 180-220 g, clean grade, provided by the university of Hunan Master.
3. Grouping experiments: the rats are adapted to be raised in a laboratory for 1 week, no abnormity exists in activity, food and excrement, and 56 rats are randomly divided into 6 groups according to a random number table: (1) normal control group: diet regular for 12 weeks; (2) alcoholic fatty liver disease model group: performing intragastric administration with 50% alcohol, 10ml/kg/d, 8 weeks; (3) compound methionine choline treatment group (10 mg/kg): after 8 weeks of high fat diet, changing into conventional diet for 4 weeks, and adding compound choline methionine (10mg/kg) into drinking water; (4) resveratrol treatment group (10 mg/kg): after 8 weeks of high fat diet, change to regular dietAdding resveratrol (10mg/kg) into drinking water for 4 weeks; (5) resveratrol A Ring N (CH)3)2Group derivative low dose treatment group (2 mg/kg): after 8 weeks of high fat diet, changing into regular diet for 4 weeks, and adding resveratrol A-ring N (CH) into drinking water3)2A base derivative (2 mg/kg); (6) resveratrol A Ring N (CH)3)2Group treated with medium dose of derivative (10 mg/kg): after 8 weeks of high fat diet, changing into regular diet for 4 weeks, and adding resveratrol A-ring N (CH) into drinking water3)2A base derivative (10 mg/kg); (7) resveratrol A Ring N (CH)3)2Group derivative high dose treatment group (50 mg/kg): after 8 weeks of high fat diet, changing into regular diet for 4 weeks, and adding resveratrol A-ring N (CH) into drinking water3)2The base derivative (50 mg/kg). Injecting the rats into the abdominal cavity with 2% sodium pentobarbital (45mg/kg) at the end of 12 weeks, taking blood from the inferior vena cava after anesthesia, centrifuging the blood specimen and detecting serological indexes; taking the left liver leaf, fixing with 10% neutral formalin solution, embedding in paraffin, and slicing for histopathological observation of liver; taking the right lobe of the liver, extracting protein and carrying out WesternBlot detection; taking liver right leaf, freezing, and performing immunofluorescence detection.
4. The experimental contents are as follows:
4.1 general conditions in rats: the activity, diet, fur gloss, and sleep status of the rats were observed.
4.2 body weight, liver index: weighing the weight by a balance, and weighing the liver after the dead anesthesia; liver index (%) ═ liver weight (g)/body weight (g) × 100%.
4.3 serum TG, TC, HDL-C content: collecting blood from rat femoral artery, collecting blood sample with heparin anticoagulation tube, centrifuging at 4 deg.C for 8 min at 1500 r/min within 30min, collecting supernatant, and freezing at-80 deg.C. Adding physiological saline into the reserved tissue according to the mass ratio of 1:9, homogenizing for 10 minutes by an electronic homogenizer, centrifuging for 8 minutes at the temperature of 4 ℃ at 1500 r/min, and taking the supernatant for freezing and storing at the temperature of-80 ℃ after centrifugation. The Elisa kit stored at 4 ℃ was allowed to equilibrate at room temperature for 30 minutes, the desired Elisa reaction plate was removed from the aluminum foil bag, and the remaining reaction plates were sealed with a self-sealing bag and stored at 4 ℃. And arranging blank holes, standard substance holes and sample holes by using a marking pen, wherein the blank holes are vacant, and 50 mu L of standard substances with different samples to be detected and different concentrations are added into the sample holes and the standard substance holes respectively. 100 mu L of detection antibody marked by horseradish peroxidase is respectively added into the sample hole and the standard product hole, and each reaction hole on the Elisa reaction plate is sealed by a sealing plate membrane and then put into a water bath kettle at 37 ℃ for incubation for 60 minutes. The liquid in the reaction well was discarded, and the residual water in the reaction well was patted dry on absorbent paper. Adding 350 mu L of washing solution prepared by distilled water and washing buffer solution according to the volume ratio of 1:20 into each reaction hole, standing for 1 minute at room temperature, throwing off the washing solution, patting dry the residual water in the reaction holes on absorbent paper, and repeating for 5 times. 50. mu.L of each substrate A, B was added to each reaction well and incubated in an incubator at 37 ℃ for 15 minutes in the absence of light. Adding 50 μ L of stop solution into each reaction well, and measuring OD value of each well at the set wavelength of 450nm in an enzyme-labeling instrument within 15 minutes. And (4) taking the concentration value of the measured standard product as a vertical coordinate and the OD value as a horizontal coordinate, drawing a standard curve by software and obtaining a linear regression equation. And substituting the OD value of the detection sample into the linear regression equation to calculate the concentration of the detection sample.
4.4 serum ALT, AST content: the method is the same as 4.3.
4.5 serum SOD, MDA content: the method is the same as 4.3.
4.6HE staining: placing the paraffin sections in an oven to bake for 1-2 hours at 60 ℃; slicing paraffin into normal xylene, and dewaxing the xylene with ethanol to water; hematoxylin staining for 10 minutes; flushing with running water to remove residual color; 0.7% ethanol hydrochloride was differentiated for several seconds; rinsing with running water, and turning the slices blue for about 15 minutes; 7.95% ethanol for 30 seconds; staining with alcoholic eosin for 30 seconds; i95% ethanol for 30 seconds; II 95% ethanol for 30 seconds; i100% ethanol for 30 seconds; II 100% ethanol for 30 seconds; xylene carbolate for 30 seconds; i xylene for 30 seconds; II xylene for 30 seconds; and (5) sealing the neutral gum.
4.7 immunohistochemistry: paraffin section is dewaxed to water; 3% H2O2Incubating for 5-10 minutes at room temperature to eliminate the activity of endogenous peroxidase; washing with distilled water, and soaking in PBS for 2 times, each for 5 min; sealing with 5-10% normal goat serum (PBS diluted), incubating for 10 minutes at room temperature, pouring off the serum, and not washing; adding primary antibody working solution dropwise, incubating at 37 deg.C for 1 hr or incubating at 4 deg.CAt night; PBS washing, 3 times, each time for 5 minutes; dripping a proper amount of biotin-labeled secondary antibody working solution, and incubating for 10 minutes at 37 ℃; PBS washing, 3 times, each time for 5 minutes; dropwise adding a proper amount of horseradish enzyme or alkaline phosphatase labeled streptavidin working solution, and incubating for 10 minutes at 37 ℃; PBS washing, 3 times, each time for 5 minutes; developing with developer for 3 min (DAB or NBT/BCIP), washing with tap water, re-dyeing, dewatering, transparentizing, and sealing.
4.8 WesternBlot: PMSF (1:100) and Cocktail (1:1000) were added to the tissue homogenate, the mixed tissue homogenate was added at a ratio of 15. mu.L/mg, and the electric homogenizer was adjusted to 30 times/mim, followed by 50 homogenations clockwise and 50 homogenations counterclockwise. The tissue suspension was pipetted into a 5mL centrifuge tube and 1/3 volumes of Buffer were added and mixed well with shaking. Then placing the mixture on a test tube rack, boiling the mixture in water bath for 10 minutes, then carrying out ultrasonic oscillation for 20 times by using 20 KHz, centrifuging the mixture, taking supernate after 4 ℃/12000g/5 minutes, and storing the supernate in a centrifuge tube at-80 ℃. The BSA gradient solution was prepared by first preparing 10. mu.g/. mu.L BSA buffer solution, and then diluting the buffer solution with double distilled water in a gradient manner to obtain a standard protein, wherein the dilution concentrations were 0. mu.g/. mu.L, 0.2. mu.g/. mu.L, 0.4. mu.g/. mu.L, 0.6. mu.g/. mu.L, 0.8. mu.g/. mu.L, and 1.0. mu.g/. mu.L, in this order. The protein supernatant centrifuge tube was taken out from a-80 ℃ freezer and placed on ice, after slow dissolution 2. mu.L of protein sample was aspirated and 18. mu.L of double distilled water was added, and mixed, shaken and mixed well. First, 3 parallel wells were set on a 96-well plate, and then the mixed protein sample and BSA gradient buffer solution were mixed and shaken as described above at a volume of 5. mu.L/well. Taking 50 parts of solution A and 1 part of solution B in the kit, mixing uniformly to prepare working solution, quickly adding 95 mu L of working solution into each hole of a 96-hole plate, putting the 96-hole plate into an incubator, and incubating for 30 minutes at 37 ℃. And setting the spectrophotometer at a 562nm light wave band wave, measuring the absorbance value, drawing a standard protein curve, and calculating to obtain the protein concentration value of the sample. Taking 1 part of bromophenol blue and 3 parts of beta-mercaptoethanol, mixing, shaking and uniformly mixing. Diluting the mixed solution by 10 times with double distilled water, boiling in a water bath for 10 minutes, and shaking and mixing uniformly. Adding 10% separation gel into the gel plate, standing at room temperature, pouring out the water layer after the gel is solidified, and sucking with filter paper. Then 5% concentrated glue is added for standing at room temperature, and the mixture is solidified for later use. Placing the rubber plate in an electricFixing in an electrophoresis tank, slowly adding an electrophoresis solution along the electrophoresis tank, fully and uniformly mixing the sample, adding a protein sample into a lane of the electrophoresis tank by using a microsyringe, carrying out electrophoresis for 30 minutes by using 80V constant voltage until the sample runs to separation gel, carrying out electrophoresis separation by using 120V constant voltage, and determining the electrophoresis time according to the molecular weight of the target protein. The marker pen marks the nitrocellulose membrane, then the nitrocellulose membrane is soaked in the pretreated membrane transferring working solution, the separation gel is peeled off, the gel is transversely cut according to the molecular weight range of the target protein, and the fiber sponge cushion → the 3-layer filter paper → the gel plate → the nitrocellulose membrane → the 3-layer filter paper → the fiber sponge cushion are sequentially placed from the negative electrode to the positive electrode. The materials are stacked into a trapezoid shape and then placed into a membrane transferring groove which is incubated at 0 ℃ in advance, membrane is transferred at a constant current of 252mA, and the membrane transferring time is determined according to the molecular weight of the target protein and is generally controlled to be 1 h. After the completion of the membrane transfer, the nitrocellulose membrane was taken out from the membrane transfer tank, and washed with PBS 2 times for 5 minutes each. Then, 100. mu.L of 10% goat serum or 2% bovine serum albumin was used for immersion at room temperature for 10 minutes, and the solution was washed 3 times with PBS for 5 minutes each. Diluting the concentrated solution at a proper ratio to 1cm240 μ L of primary antibody was added dropwise. Incubating at 37 ℃ for 1-2 h or standing overnight at 4 ℃, washing with a washing buffer solution for 3 times, washing for 10 minutes each time → taking out the nitrocellulose membrane on the next day and recovering the primary antibody, adding 0.02% Tween 20 into the TBS buffer solution, rinsing for 3 times, rinsing for 5 minutes each time, sealing with a freshness protection package, selecting a concentrated solution diluted in a proper proportion and then carrying out 1cm 240 μ L of biotinylated secondary antibody was added dropwise, incubated at 37 ℃ for 1h or at 4 ℃ overnight, washed with PBS for 5 min and repeated 3 times. The nitrocellulose membrane was removed and the secondary antibody recovered, and 0.02% tween 20 was added to the TBS buffer and rinsed 3 times for 5 minutes each. After rinsing, rinse 3 times 5 minutes each with TBS buffer. The nitrocellulose membranes were scanned at the 700nm light wave band, respectively.
4.9 immunofluorescence: incubating the frozen section with the antibody, and washing with PBS for 3 times, 5 minutes each time; diluting primary antibody according to corresponding proportion, dripping 50 μ L of primary antibody into each hole, incubating overnight at 4 ℃, recovering the primary antibody, washing with PBS, and multiplying by 3 times for 5 minutes; dripping fluorescent secondary antibody of corresponding species, incubating for 1 hour at room temperature in a dark place, removing the secondary antibody, and washing with PBS for 3 times, 5 minutes each time; adding the diluted second primary antibody dropwise, 50 mu L/hole, and incubating overnight at 4 ℃; primary antibody is recovered, PBS is washed for 3 times, and each time lasts for 5 minutes; and (3) dropwise adding fluorescent secondary antibody of the corresponding species, incubating for 1 hour at room temperature in a dark place, removing the secondary antibody, washing with PBS for 3 times, and dropwise adding a small amount of PBS every 5 minutes to cover the cells. 0.1% TritonX-100 was added to the PBS buffer, and the 96-well plate was washed 3 times for 5 minutes each, and PBS diluted at 1:500 times with Hoechst was added and left for 10 minutes in the dark. 0.1 percent TritonX-100 is added into PBS buffer solution, the 96-hole culture plate is washed for 3 times, each time is 3 minutes, 100 mu L of fluorescence-resistant quenching mounting solution is dripped on a glass slide and mounted by a cover slip, the glass slide is kept at the room temperature for 1 minute, and then the horizontal position is kept and placed in a refrigerator at 4 ℃ and is kept away from light. After 24 hours, the cell morphology and the staining condition can be observed under a fluorescence microscope or a laser confocal microscope.
5. The statistical method comprises the following steps: all data are expressed as means ± standard deviation (± s). Difference comparison between groups was performed by ANOVA and Newman-Student multiple comparisons; the analysis of t test is completed by SPSS 13.0 statistical software, and the difference is considered to be significant when P on two sides is less than 0.05.
6. Results
6.1 general case: (1) the rats in the normal control group are full of energy, flexible and well-moving, have normal diet, clean and tidy fur and slowly increase the weight; (2) the alcoholic fatty liver disease model group has large food consumption, sleepiness, yellowish hair, larger individuals, faster weight increase than the normal group, listlessness, reduced activity and messy fur; (3) the compound methionine choline treatment group and the resveratrol treatment group slightly improve or do not obviously improve the symptoms; (4) resveratrol A Ring N (CH)3)2Low dose treatment group of resveratrol A ring N (CH) based derivatives3)2Resveratrol A Ring N (CH) in Medium dose treatment group of the derivatives3)2The symptoms of the high-dose treatment group of the derivative are obviously improved, and the improvement degree is gradually enhanced along with the increase of the dose.
6.2 body weight, liver index: the results are shown in Table 1 and FIGS. 3-5, FIG. 3 is a graph of the effect of different experimental groups on the quality of the initial body, wherein a P is compared to the normal control group<0.01; b P comparison with alcoholic fatty liver disease model group<0.01; compared with the compound methionine choline treatment group,c P<0.01; d P comparison with resveratrol treatment group<0.01. FIG. 4 is a graph of the effect of different experimental groups on end body mass, wherein a P is compared to a normal control group<0.01; b P comparison with alcoholic fatty liver disease model group<0.01; c P in comparison with Compound methionine Choline treatment group<0.01; d P comparison with resveratrol treatment group<0.01. FIG. 5 is a graph of the effect of different experimental groups on liver index, wherein a P is compared to a normal control group<0.01; b P comparison with alcoholic fatty liver disease model group<0.01; c P in comparison with Compound methionine Choline treatment group<0.01; dP compared with resveratrol treatment group<0.01. As shown in Table 1 and FIGS. 3 to 5, (1) compared with the normal control group, the initial body mass of rats in other groups has no obvious difference (P is more than 0.01); (2) resveratrol A Ring N (CH) compared to alcoholic fatty liver disease model group3)2Resveratrol A Ring N (CH) in Medium dose treatment group of the derivatives3)2The terminal body mass and the liver index of rats in the high-dose treatment group of the derivative are obviously reduced (P)<0.01); (3) resveratrol A Ring N (CH) compared to resveratrol treatment group3)2Resveratrol A Ring N (CH) in Medium dose treatment group of the derivatives3)2The terminal body mass and the liver index of rats in the high-dose treatment group of the derivative are obviously reduced (P)<0.01)。
Table 1 rat body weight, liver index (n-8,
)
note: aP <0.01 compared to normal control group; bP <0.01 compared to alcoholic fatty liver disease model group; cP <0.01 compared to the compound methionine choline treatment group; dP <0.01 compared to resveratrol treatment group.
6.3 serum TG, TC, HDL-C content: the results are shown in Table 2 and FIGS. 6-8, FIG. 6 is the effect of different experimental groups on the content of TG in serum, compared with the normal control group, a P<0.01; compared with alcoholic fatty liver disease model groupRelatively, b P<0.01; c P in comparison with Compound methionine Choline treatment group<0.01; d P comparison with resveratrol treatment group<0.01. FIG. 7 is a graph of the effect of different experimental groups on serum TC levels, compared to a normal control group, a P<0.01; b P comparison with alcoholic fatty liver disease model group<0.01; c P in comparison with Compound methionine Choline treatment group<0.01; d P comparison with resveratrol treatment group<0.01. FIG. 8 is a graph of the effect of different experimental groups on serum HDL-C levels, compared to a normal control group, a P<0.01; b P comparison with alcoholic fatty liver disease model group<0.01; c P in comparison with Compound methionine Choline treatment group<0.01; d P comparison with resveratrol treatment group<0.01. As shown in Table 2 and FIGS. 6 to 8, (1) resveratrol A-ring N (CH) in comparison with the alcoholic fatty liver disease model group3)2Resveratrol A Ring N (CH) in Medium dose treatment group of the derivatives3)2The TG and TC of rats in the high-dose treatment group of the derivative are both obviously reduced, and HDL-C is obviously increased (P)<0.01); (2) resveratrol A Ring N (CH) compared to resveratrol treatment group3)2Resveratrol A Ring N (CH) in Medium dose treatment group of the derivatives3)2The TG and TC of rats in the high-dose treatment group of the derivative are both obviously reduced, and HDL-C is obviously increased (P)<0.01)。
Table 2 serum TG, TC, HDL-C content (n-8,
)
note: aP <0.01 compared to normal control group; bP <0.01 compared to alcoholic fatty liver disease model group; cP <0.01 compared to the compound methionine choline treatment group; dP <0.01 compared to resveratrol treatment group.
6.4 serum ALT, AST content: the results are shown in Table 3 and FIGS. 9-10, FIG. 9 is the effect of different experimental groups on serum ALT content, compared with normal control group, a P<0.01; b P comparison with alcoholic fatty liver disease model group<0.01; andcomparison of Compound methionine Choline treatment group, c P<0.01; d P comparison with resveratrol treatment group<0.01. FIG. 10 is a graph of the effect of different experimental groups on serum AST levels, compared to a normal control group, a P<0.01; bP compared with alcoholic fatty liver disease model group<0.01; c P in comparison with Compound methionine Choline treatment group<0.01; d P comparison with resveratrol treatment group<0.01. As can be seen from Table 3 and FIGS. 9 to 10: (1) resveratrol A Ring N (CH) compared to alcoholic fatty liver disease model group3)2Low dose treatment group of resveratrol A ring N (CH) based derivatives3)2Resveratrol A Ring N (CH) in Medium dose treatment group of the derivatives3)2The ALT and AST of the rats in the high-dose treatment group of the derivative are both obviously reduced (P)<0.01); (2) resveratrol A Ring N (CH) compared to resveratrol treatment group3)2Resveratrol A Ring N (CH) in Medium dose treatment group of the derivatives3)2The ALT and AST of the rats in the high-dose treatment group of the derivative are both obviously reduced (P)<0.01)。
Table 3 serum ALT, AST content (n-8,
)
note: a P < 0.01; b P <0.01 compared to the alcoholic fatty liver disease model group; c P <0.01 compared to the compound choline methionine treatment group; d P <0.01 compared to the resveratrol treatment group.
6.5 serum SOD, MDA content: the results are shown in Table 4 and FIGS. 11-12, FIG. 11 is the effect of different experimental groups on serum SOD content, compared with normal control group, a P<0.01; b P comparison with alcoholic fatty liver disease model group<0.01; c P in comparison with Compound methionine Choline treatment group<0.01; d P comparison with resveratrol treatment group<0.01. FIG. 12 is a graph showing the effect of different experimental groups on the MDA content in serum, compared to a normal control group, a P<0.01; comparison with alcoholic fatty liver disease model group，b P<0.01; c P in comparison with Compound methionine Choline treatment group<0.01; d P comparison with resveratrol treatment group<0.01. As can be seen from Table 4 and FIGS. 11 to 12: (1) compared with alcoholic fatty liver disease model group, resveratrol treatment group, resveratrol A ring N (CH)3)2Low dose treatment group of resveratrol A ring N (CH) based derivatives3)2Resveratrol A Ring N (CH) in Medium dose treatment group of the derivatives3)2SOD and MDA (P) of rats treated with high dose of the derivative are obviously increased and MDA is obviously reduced<0.01); (2) compared with resveratrol treatment group, SOD and MDA are in resveratrol A ring N (CH)3)2There was no difference between the dose groups of the base derivative (P > 0.01). The key points are as follows: resveratrol has good antioxidation, but has no good intervention effect on alcoholic fatty liver; resveratrol A Ring N (CH)3)2The derivative has good antioxidation, but has good intervention effect on alcoholic fatty liver. In particular, resveratrol A Ring N (CH)3)2The intervention of the derivatives on alcoholic fatty liver is not accomplished by antioxidant action.
Table 4 serum SOD, MDA content (n ═ 8,
)
note: a P < 0.01; b P <0.01 compared to the alcoholic fatty liver disease model group; c P <0.01 compared to the compound choline methionine treatment group; d P <0.01 compared to the resveratrol treatment group.
6.6HE staining: the results are shown in fig. 13, and fig. 13 shows the effect of different experimental groups on the structure under the liver microscope (400 ×), and it can be seen that (1) the hepatic cells of the rat in the alcoholic fatty liver disease model group have obvious balloon-like changes, and the fatty degeneration area is obviously higher than that of the normal control group; (2) the resveratrol treatment group has no obvious inhibition effect on ballooning of liver cells, and has fatThe variable area is not obviously reduced; (3) resveratrol A Ring N (CH)3)2Resveratrol A Ring N (CH) in Medium dose treatment group of the derivatives3)2The liver cells of rats treated by high dose of the derivative have obvious balloon-like changes, and the fatty degeneration area is obviously higher than that of rats of alcoholic fatty liver disease model groups.
6.7 immunohistochemistry: the results are shown in FIG. 14, and FIG. 14 shows the effect of different experimental groups on immunohistochemistry for expression of NHE-1 in liver (400X), which indicates that (1) the liver cells of rats in the alcoholic fatty liver disease model group have significantly increased yellowish-brown stained granules, and the expression of NHE1 is significantly higher than that of the normal control group; (2) the resveratrol treatment group does not inhibit the increase of yellow-brown staining particles of liver cells, and the expression of NHE1 is not obviously different from that of the alcoholic fatty liver disease model group; (3) resveratrol A Ring N (CH)3)2Resveratrol A Ring N (CH) in Medium dose treatment group of the derivatives3)2The high-dose treatment group of the derivative obviously inhibits the increase of yellow-brown stained granules of the liver cells, and the expression of NHE1 is obviously different from that of the alcoholic fatty liver disease model group.
6.8 WesternBlot: the results are shown in FIGS. 15-16, FIG. 15 is the effect of different experimental groups on hepatic NHE-1 expression of WesternBlot (400 ×), wherein (1) is a normal control group; (2) alcoholic fatty liver disease model group; (3) compound methionine choline treatment group; (4) a resveratrol treatment group; (5) resveratrol A Ring N (CH)3)2A group of low dose treatments with the derivatives; (6) resveratrol A Ring N (CH)3)2A medium dose treatment group of the derivative; (7) resveratrol A Ring N (CH)3)2Group treated with high dose of the base derivative. FIG. 16 is a statistical plot of the effect of different experimental groups on hepatic NHE-1 expression of WesternBlot, compared to a normal control group, a P<0.01; b P comparison with alcoholic fatty liver disease model group<0.01; c P in comparison with Compound methionine Choline treatment group<0.01; d P comparison with resveratrol treatment group<0.01. As can be seen from FIGS. 15 to 16: (1) the hepatic cell gray scale of the rat in the alcoholic fatty liver disease model group is obviously increased, and the expression of NHE1 is obviously higher than that of the normal control group; (2) the resveratrol treatment group does not inhibit the gray scale increase of liver cells, NHE1 expression and alcoholic fatty liver diseaseNo significant difference was observed in the model groups; (3) resveratrol A Ring N (CH)3)2Resveratrol A Ring N (CH) in Medium dose treatment group of the derivatives3)2The high-dose treatment group of the derivative obviously inhibits the gray scale increase of the liver cells, and the expression of NHE1 is obviously different from that of the alcoholic fatty liver disease model group.
6.9 immunofluorescence: the results are shown in fig. 17, fig. 17 is a graph of the effect of different experimental groupings on hepatic PI3K and STAT immunofluorescence (400 ×), wherein (1) normal control group; (2) alcoholic fatty liver disease model group; (3) compound methionine choline treatment group; (4) a resveratrol treatment group; (5) resveratrol A Ring N (CH)3)2A group of low dose treatments with the derivatives; (6) resveratrol A Ring N (CH)3)2A medium dose treatment group of the derivative; (7) resveratrol A Ring N (CH)3)2Group treated with high dose of the base derivative. As can be seen from fig. 17: (1) the rat liver cell PI3K and STAT fluorescence of the alcoholic fatty liver disease model group are obviously enhanced, and the expression is obviously higher than that of a normal control group; (2) the fluorescence enhancement of PI3K and STAT is not inhibited in the resveratrol treatment group, and the expression is not obviously different from that in the alcoholic fatty liver disease model group; (3) resveratrol A Ring N (CH)3)2Resveratrol A Ring N (CH) in Medium dose treatment group of the derivatives3)2The high-dose treatment group of the derivative obviously inhibits the fluorescence enhancement of the hepatic cell PI3K and STAT, and the expression of the derivative is obviously different from that of an alcoholic fatty liver disease model group.
The foregoing is merely a preferred embodiment of the invention and is not intended to limit the invention in any manner. It should be noted that, for those skilled in the art, without departing from the principle of the present invention, several improvements and modifications can be made, and these improvements and modifications should also be construed as the protection scope of the present invention.
Claims (10)
1. Resveratrol A ring N (CH)3)2A group derivative having the structure shown in formula I:
2. resveratrol A ring N (CH) as in claim 13)2The preparation method of the derivative is characterized by comprising the following steps:
mixing diethyl phosphite, an organic solvent, NaH and 4-methoxybenzyl bromide to carry out debromination reaction to obtain a compound with a structure shown in a formula 1;
mixing 4-dimethylamino salicylaldehyde, an organic solvent, chloromethyl methyl ether and diisopropylethylamine for a substitution reaction to obtain a compound with a structure shown in a formula 2;
mixing a compound with a structure shown in a formula 1, a compound with a structure shown in a formula 2, an organic solvent and NaH for a condensation reaction to obtain a compound with a structure shown in a formula 3;
mixing the compound with the structure shown in the formula 3, tert-butyl alcohol and pyridinium p-toluenesulfonate, and refluxing to obtain resveratrol A ring N (CH) with the structure shown in the formula I3)2A derivative of phenyl;
3. the preparation method according to claim 2, wherein the debromination reaction time is 1-3 hours.
4. The method according to claim 2, wherein the time for the substitution reaction is 10 to 30 min.
5. The method according to claim 2, wherein the condensation reaction is carried out at a temperature of 0 to 100 ℃ for 1 to 3 hours.
6. The preparation method according to claim 2, wherein the reflux temperature is 90-100 ℃ and the reflux time is 1-3 h.
7. The preparation method according to claim 2, wherein the molar ratio of the compound having the structure represented by formula 3 to the pyridinium p-toluenesulfonate is 1:2 to 2.1.
8. Resveratrol A ring N (CH) as in claim 13)2The derivative or resveratrol A ring N (CH) prepared by the preparation method of any one of claims 2-73)2The application of the derivative in preparing medicines for treating alcoholic fatty liver is provided.
9. The use of claim 8, wherein the medicament for treating alcoholic fatty liver disease comprises an effective amount of resveratrol A ring N (CH) having a structure shown in formula I3)2The derivative, the stereoisomer and the pharmaceutically acceptable salt thereof and pharmaceutically acceptable carriers, auxiliary materials, excipients and diluents.
10. The use of claim 8, wherein the dosage form of the drug for treating alcoholic fatty liver disease comprises a pharmaceutically acceptable dosage form of tablet, injection, capsule, granule, pill, powder, oral liquid, sustained release preparation, controlled release preparation or nano preparation.
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Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO1993023357A1 (en) * | 1992-05-21 | 1993-11-25 | Research Corporation Technologies, Inc. | Stilbene derivatives as anticancer agents |
| US20040152629A1 (en) * | 2000-12-21 | 2004-08-05 | Hadfield John Anthony | Substituted stilbenes and their reactions |
| US20070249647A1 (en) * | 2006-04-25 | 2007-10-25 | Vander Jagt David L | Substituted CIS- and trans-stilbenes as therapeutic agents |
| CN104147026A (en) * | 2014-08-20 | 2014-11-19 | 安树君 | New application of resveratrol derivative |
-
2020
- 2020-08-20 CN CN202010841450.9A patent/CN111875513B/en active Active
Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO1993023357A1 (en) * | 1992-05-21 | 1993-11-25 | Research Corporation Technologies, Inc. | Stilbene derivatives as anticancer agents |
| US20040152629A1 (en) * | 2000-12-21 | 2004-08-05 | Hadfield John Anthony | Substituted stilbenes and their reactions |
| US20070249647A1 (en) * | 2006-04-25 | 2007-10-25 | Vander Jagt David L | Substituted CIS- and trans-stilbenes as therapeutic agents |
| CN104147026A (en) * | 2014-08-20 | 2014-11-19 | 安树君 | New application of resveratrol derivative |
Non-Patent Citations (2)
| Title |
|---|
| MARK CUSHMAN等: "Synthesis and Evaluation of Stilbene and Dihydrostilbene Derivatives as Potential Anticancer Agents That Inhibit Tubulin Polymerization", 《J. MED. CHEM.》 * |
| SOO SUNG KANG等: "Synthesis and biological evaluation of a library of resveratrol analogues as inhibitors of COX-1, COX-2 and NF-κB", 《BIOORGANIC & MEDICINAL CHEMISTRY》 * |
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