CN101543489B - Compounds as Liver X Receptor Modulators - Google Patents
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Abstract
本发明公开了一类作为肝X受体调节剂的化合物，具体而言，涉及一类白藜芦醇四聚体化合物，这类化合物能用于制备治疗或预防与肝X受体(LXR)活性相关的疾病或病症的药物，包括预防和治疗神经退行性疾病、心血管疾病、糖尿病、高胆固醇血症或肥胖症。 The invention discloses a class of compounds as liver X receptor modulators, in particular, relates to a class of resveratrol tetramer compounds, which can be used for the preparation of treatments or prevention of liver X receptor (LXR) Drugs for activity-related diseases or conditions, including prevention and treatment of neurodegenerative diseases, cardiovascular diseases, diabetes, hypercholesterolemia or obesity. 
Description
Technical field
The invention provides the application of resveratrol tetramerization compound in the medicine for preparing treatment or the prevention disease relevant with liver X receptor (LXR) activity or disease, comprise prevention and treatment neurodegenerative diseases, cardiovascular disease, diabetes, hypercholesterolemia or obesity.
Background technology
Liver X receptor (Liver X receptor, LXR) be the transcription factor that belongs to nuclear receptor superfamily, lxr receptor is by being connected to the upper heterodimer that forms of dimension first X receptor (RXR), and this heterodimer is with ad hoc fashion and DNA response element (LXRE) combination (the Genes dev.1995 that causes the target gene transcription activating; 9:1033-45).Can utilize LXR and (or) the RXR part activates described LXR/RXR dimer.The simultaneously transcription activating of target gene requirement is additional, and (Nature 1996,383:728-31) such as the co-activations such as SRC-1.
This receptor relates to many metabolic pathways, particularly relates to the homoiostasis of cholesterol, bile acid, triglyceride and glucose.(World Chin J Digestol 2005，28：13(16))
Confirmed at present two types lxr receptor (being LXR α and LXR β), LXR β generally expresses in vivo, LXR α expresses in liver and (Gene 2000 with less degree expression at kidney, intestinal, fatty tissue and spleen, 243:93-103, N.Y.Acad.Sci.1995,761:38-49).
Although cholesterol does not directly activate LXR, its single oxygen derivant (oxidation sterin) is such as 22-(R)-hydroxyl-cholesterol, 24-(S)-hydroxy cholesterol and 24 (S), 25-epoxy cholesterol.These oxidation sterin are that LXR is in physiological ligand (Nature 1996,383:728-31, J.Biol.Chem.1997:272:3137-40)
In liver, the identification of LXR high expressed and its endogenic ligand has shown its requisite effect in cholesterol metabolism.Under physiological condition, the homoiostasis of cholesterol can be kept by the adjusting of de novo synthesis and catabolism passage.Via the feedback mechanism that comprises the transcription factor such as LXR regulation and control target gene SREBP-1 and SREBP-2, (Cell 1997,89:331-340) in the inhibition that steroid gathering in liver can cause Biosynthesis of cholesterol.Rate-limiting enzyme-cholesterol 7α-hydroxylase (CYP7A) promotes from cholesterol to 7 Alpha-hydroxies LXR in the bile acid biosynthesis approach by raising in liver-conversion (J.Biol.Chem.1997,272:3137-3140) of cholesterol.By using LXR α shortage property mice, proved that LXR relates to bile acid biosynthesis, and therefore related to the homeostatic adjusting of cholesterol, when described mice is fed rich fatty diet, gather a large amount of cholesteryl esters (Cell 1998,93:693-704) edema caused by disorder of the liver is flat
LXR can be incorporated into ATP in conjunction with box protein-1 (ABCA1) promoter and increase gene expression to produce ABCA1 albumen, ABCA1 is as a kind of film binding transport albumen, it participates in cholesterol regulating cell drain outside liver and relate to the control of the cholesterol antiport that extra cholesterol is exported from peripheral tissues to liver to newborn high density lipoprotein (HDL) granule.Cholesterol can be adsorbed through apoAI (apolipoprotein) and ABCA1 by pre-bHDL (high density lipoprotein precursor), thereby is transported to liver, and the cholesterol metabolism that is decomposed is drained behind the bile acid in liver.The ABCA1 gene mutation causes low-level HDL and follows cardiovascular disease such as the danger of atherosclerosis, myocardial infarction and ischemic stroke increase (Nat.Genet.1999; 22:336-345).The agonist of LXR has shown increases ABCA1 gene expression, increases thus the HDL cholesterol, thereby reduces simultaneously the clean absorption of cholesterol and the risk of reduction cardiovascular disease.People's macrophage is to the load of cholesterol and the activation of lxr receptor, the expression of ABCA1 and the outflow (Biochem.Biophys.Comm.1999 of cholesterol have been induced, 257:29-33), confirm also that subsequently LXR can also regulate the expression of abc transport body family member on the intestinal level such as ABCG1, ABCG5 and ABCG8 by forming the RXR/LXR heterodimer, promote the Cholesterol influx enteric cavity and suppress the absorption of cholesterol, thereby reduce the content of blood cholesterol.(J.Biol.Chem.2000，275：14700-14707，Proc.Natl.Acad.Sci.USA.2000；97：817-822，J.Biol.Chem.2002，277：18793-18800，Proc.Natl.Acad.Sci.USA2002，99：16237-16242，Molecular Endocrinology 17(6)：985-993)
Lxr receptor also relates to the adjusting that apo E (ApoE) is expressed.The hepatic clearance of this protein and lipoprotein has very strong correlation and promote that cholesterol flows out from the macrophage that is rich in lipid.Verified by being arranged in the LXR response element (LXRE) of ApoE promoter sequence, increase (the Proc.Natl.Acad.Sci that the activation of lxr receptor can cause ApoE to express, USA2001,98:507-512), also confirm the LXR part two kinds of different murine models (ApoE-/-mice and LDLR-/-mice) in can reduce atherosclerotic damage.(Proc.Natl.Acad.Sci.USA2002,99:7604-7609, FEBS Letters 2003,536:6-11), these results show that LXR part and agonist can consist of the therapeutic agent of atherosclerosis (AS).
The cholesterol homoiostasis plays a key effect in central nervous system is unified neural degeneration mechanism equally, thus in isolated primary neuron from the Mus embryo, star spongiocyte and microglia culture the expression to described ABCA1 be studied.And confirmed abundant expression the in the star spongiocyte of ApoE in brain, the micromolecule part of LXR can significantly increase the expression of ApoE in people's star spongiocyte system (CCF-STGG1), and lxr agonist can be induced expression and the secretion of ApoE equally on the star spongiocyte of former culture.But the expression of ApoE in hippocampus and the cerebral cortex in the micromolecule agonist T0901317 up regulation body of LXR.Because ApoE plays an important role in lipoprotein removing and cholesterol redistribution, and lxr agonist can be regulated the expression of ABCA1 simultaneously, can participate in effluxing of cholesterol in the brain, therefore can improve the central nervous system relevant with the cholesterol homoiostasis neurodegenerative disease of unifying, such as Alzheimer.Result of study confirms that the LXR activation can cause that amyloid beta produces and the minimizing of secretion, thereby can cause minimizing (Journal of Neurochemistry, 2004,88 (3): the 623-634 of amylaceous precipitation in the brain.J.Biol.Chem.2003，275(15)/；13244-13256 J.Biol.Chem.2003，278(30)：27688-27694，J.Biol.Chem.2005，280(6)：4079-4088)。
It also is known (Nature Medecine 2003,9:213-219, J CliInvest, 2006,116 (3): 607-614) that lxr receptor relates to inflammatory process.Macrophage particularly plays an important role in the atherosclerotic pathogenesis in inflammation.Verified, the activation of described LXR can suppress to relate to the expression of gene on the macrophage level of inflammation.External, suppressed the expression of media such as nitricoxide synthase, cyclo-oxygenase-2 (COX-2), interleukin-1 ' beta ' (IL-1 β), interleukin-6 (IL-6), in vivo, lxr agonist has also alleviated the inflammation in the Dermatite model and has suppressed to relate to the expression of atherosclerosis mouse aorta inflammation related gene.Verified, lxr agonist can also suppress the expression of inflammation related gene in the astrocyte and microglia in the Mus brain (Journal of Neuroimmunology 2007,183:50-59)
By the adjusting that CETP (cetp) is expressed, the activation of lxr receptor also promotes the counter transport of cholesterol, described CETP relates to the transfer (J.Clin.Invest.2000,513-520) of cholesterol from the HDL lipoprotein to the lipoprotein that is rich in triglyceride of draining by liver of esterification.
In addition, confirmed that also LXR has important function in the carbohydrate metabolism process, can suppress the expression of glyconeogenesis key enzyme, suppressed the glycogen heteroplasia, the output of restriction hepatic glycogen improves the glycogen utilization rate.Can promote simultaneously the genetic transcription of glucose transporter 4 (glut4), improve peripheral tissues to the picked-up of glucose.Cause the reduction of blood sugar level and Hepatic glucose production.(Diabetes 2004,53, suppl 1, S36-S42) in addition, the LXRs also aspect such as the expression of the synthetic and secretion by regulating insulin, inflammatory factor and glucocorticoid activity affects generation, development (the Intern J Endocrinol Metab of diabetes, 2006,26 (1): 43-53).As use lxr agonist can cause the blood glucose decrease to the treatment of diabetes rodent, particularly in insulin resistance Zuker. rat (fa/fa), the LXR activation has suppressed the gene relevant with gluconeogenesis.(J.Biol.Chem.2003，278(2)：1131-1136)。
Particularly based on Publication about Document, chemical compound of the present invention is that prior art is known,
1) formula (I) chemical compound: be resveratrol tetramer compound, its Hopeaphenol
2) formula (II-III) chemical compound is resveratrol tetramer compound, its 7 ' "-8 ' "-two dehydrogenation vitisin As (7-8 ' "-didehydro-vitisinA) and vitisin A (vitisinA)
Formula (I) formula (II) formula (III)
Described document has: WO 03/090869, WO 03/082192, WO 03/082802, WO 05/077122 or WO 05/121093.The described chemical compound antitumor action of formula (I) and formula (I-III) chemical compound antiinflammatory action are that prior art is known.(Phytochemistry 58 (2001) 357-362, Journal ofExperimental Therapeutics and Oncology 3:283-288,2003), but its LXR regulating action has no report.
Under this background, the lxr receptor active regulator chemical compound that searching can be used for the treatment of following particular pathologies has great importance.Be used for the treatment of LXR active relevant such as cardiovascular disease, hypercholesterolemia, dyslipidemia, myocardial infarction, atherosclerosis, diabetes, obesity and neurodegenerative diseases.
Summary of the invention
The object of the present invention is to provide the new chemical compound that can regulate the LXR activity, for resveratrol tetramer compound, its Hopeaphenol suc as formula shown in (I), resveratrol tetramer compound, its 7 ' "-8 ' "-two dehydrogenation vitisin As (7-8 ' "-didehydro-vitisinA) suc as formula shown in (II) and vitisin A (vitisinA) suc as formula shown in (III)
Formula (I) formula (II) formula (III)
The upper related activity agent of at least a and another kind for the treatment of in the chemical compound of formula (I-III) or its pharmaceutically acceptable salt is such as being combined in for the preparation of the purposes in the medicine in the treatment Animal diseases of anti-diabetic and anti-hypercholesterolemiccompounds activating agent.
Therefore, active substance of the present invention is for the preparation of the medical product take treatment as purpose, described therapeutic use is especially for the treatment of hypercholesterolemia, dyslipidemia, diabetes, obesity and cardiovascular disease, and these diseases are results of cholesterol metabolism and serum lipoprotein imbalance.The compounds of this invention also can be used as active substance take prevention or treat following disease and use in the medical product of purpose, described disease is atherosclerosis, myocardial infarction, such as the specific inflammation such as dermatitis with such as neurodegenerative diseases such as Alzheimer.
Particularly, chemical compound of the present invention is regulated the active of LXR and therefore be can be used for treating wherein LXR to pathology and/or the effective disease of symptomatology or the disease of described disease.The present invention further provides the compounds of this invention for the preparation for the treatment of LXR wherein to the purposes in the medicine of the pathology of described disease and/or the effective disease of symptomatology or disease.Disease or the sufferer of LXR mediation comprise: inflammation, cardiovascular disease, comprise atherosclerosis, hypercholesterolemia, hyperlipemia and glucose homeostasis obstacle, and comprise insulin resistant, type ii diabetes and obesity.
Lipoprotein metabolism is a kind of dynamic process composed as follows: from liver, produce be rich in triglyceride and cholesterol granule as very low density lipoprotein (VLDL) (VLDL), modify hdl particle in these blood plasma (VLDL to intermediate density (IDL) to low-density (LDL)) and from blood plasma, remove described granule by liver again.This process provides triglyceride and free cholesterol has been transported to somatic cell.The cholesterol antiport reaches the mechanism that aforesaid LXR works, and as raising the expression of ABCA1 and ApoE, in conjunction with apoAI, the cholesterol excessive by this mechanism turned back to the liver outside hepatic tissue.This process is undertaken by high density lipoprotein (HDL) cholesterol.Lipoprotein (VLDL, HDL) produces from liver, and the combination that the modification of blood plasma endoparticle (all) and subsequently removing turn back to liver is the reason that keeps cholesterol Css in the blood plasma.Chemical compound of the present invention increases the antiport of cholesterol from the outflow of peripheral cells by increasing cholesterol.Numerous results of study show that lxr agonist is by raising the expression of ABCA1 and ApoE, mechanism in conjunction with apoAI cholesterol antiport, the macrophage foam cell formation that inhibited oxidation low density lipoprotein, LDL (ox-LDL) is induced, the formation of the early stage focus of inhibition atherosclerosis, realization are disappeared and are reversed.(Prog.Biochem.Biophys.2003；30(6)：940-944)。The present invention includes the compounds of this invention for the preparation of the purposes that increases in the cholesterol antiport medicine.The invention provides in addition for the chemical compound that suppresses cholesterol absorption and the compounds of this invention and be used for suppressing the purposes of clean cholesterol absorption and the purposes of inhibition macrophage foam cell formation.
The compounds of this invention can also be used for prevention or treatment inflammation and neurodegenerative diseases or neurodegenerative disease.Therefore, the present invention also provides the method for prevention or treatment inflammation and the method for prevention or treatment neurodegenerative diseases or neurodegenerative disease, and particularly impaired the or inflammation of neuronal degeneration, neuronal damage or the plasticity in the CNS is neurodegenerative disease or the disease of feature.Therefore be characterised in that neuronal degeneration in the brain, inflammation, cholesterol and dyslipidemias and have benefited from the specified disease of neure growth and/or reparation or sufferer comprises that apoplexy, Alzheimer, volume temporo dementia (tauopathy), peripheral neurophaty, parkinson disease, the dementia with Lu Yiti (Lewybodies), amyotrophic lateral sclerosis and multiple sclerosis and Ni-Pi are sick.In addition, chemical compound of the present invention can be used for the treatment of or prevent various diseases with inflammatory component, such as rheumatoid arthritis, osteoarthritis, psoriasis, asthma etc.
Aforementioned lxr agonist improves glucose tolerance and strengthens glut4 expresses (US60/436,112; US10/745,334), and tunable is present in the collaborative adjusting of the related gene of glucose metabolism in liver and the fatty tissue.In liver, lxr agonist suppresses the expression of several genes that the hepatic glycogen heteroplasia is played an important role, peroxisome proliferator-activated receptor conactivator 1 α (PGC-1 α) for example, the expression of enolpyruvate phosphate carboxylase (PEPCK) and G-6-Pase, induce the expression of liver glucokinase, promote the utilization of hepatic glucose.Find that also lxr agonist up regulation glut4 mRNA level in the fatty tissue and the glucose uptake in external 3T3-L1 fat tissue cell are enhanced.
Based on above discovery, the invention provides the method that strengthens glut4 expression in experimenter's cell, can be undertaken by giving chemical compound of the present invention to described experimenter.The present invention also provides the method for the treatment of diabetes and associated conditions such as obesity or hyperglycemia, carries out with the symptom of improving described disease by the compounds of this invention that the experimenter is given effective dose.For example, type ii diabetes is fit to treat with method of the present invention.Because lxr agonist can strengthen cell to the sensitivity of insulin and glucose uptake, use chemical compound of the present invention and can also treat other and be characterised in that insulin malfunction (for example opposing, inactivation or shortage) and/or glucose change the disease of cell deficiency over to.
In pancreas, the LXR activation stimulates insulin secretion by the metabolism of glucose and lipid in the adjusting pancreatic beta cell, thereby has pointed out the another kind of mechanism of LXR antidiabetic effect.The LXR regulator can also be secreted from pancreas by the increase insulin thus and regulate glucose tolerance.
According to foregoing description, the present invention further provides the method for preventing or treating above-mentioned any disease or disease the experimenter of this class treatment of needs.The method comprises treats at least a chemical compound or its pharmaceutically acceptable salt or its pharmaceutical composition in (" administration and the pharmaceutical composition " part in vide infra) formula of the present invention (I-III) chemical compound of effective dose to described experimenter.For any such use, required dosage is different with administering mode, the specified disease for the treatment of and required effect.
Administration and pharmaceutical composition
Generally speaking, the compounds of this invention is to treat effective dose, to give separately or with one or more therapeutic combinations via arbitrarily commonly used and acceptable mode as known in the art.The treatment effective dose can be with the effect of the order of severity, subject age and relative health condition, the compound used therefor of disease and other factors and wide variations.Generally speaking, under every daily dose of about 0.03-2.5mg/kg body weight, obtain gratifying whole body effect.Than large mammals for example the applicable every daily dose among the people in about 0.5mg-100mg scope.
The approach that chemical compound of the present invention can pass through any conventional is as the pharmaceutical composition administration, and is particularly by gastrointestinal, for example oral, for example is tablet or capsule form; Or by parenteral, for example be Injectable solution or suspension form; The part for example is lotion, gel, ointment or cream or with nose with or suppository form.Comprise that the compounds of this invention of free form or pharmaceutically acceptable salt form and the pharmaceutical composition of at least a pharmaceutically acceptable carrier or diluent can by conventional methods, prepare through mixing, granulation or coating method.For example, Orally administered composition can be tablet or capsule, they comprise active component and: 1) diluent, for example lactose, glucose, sucrose, glycogen reveal alcohol, sorbitol, cellulose and/or glycine; 2) lubricant, for example silicon dioxide, Pulvis Talci, stearic acid, its magnesium or calcium salt and/or Polyethylene Glycol; With regard to tablet, also comprise 3) binding agent, for example aluminium-magnesium silicate, gelatinized corn starch, gelatin, methylcellulose, sodium carboxymethyl cellulose and/or polyvinylpyrrolidone; Such as needs, also have 4) disintegrating agent, for example starch, agar, alginic acid or its sodium salt or effervescent mixture; And/or 5) absorbent, coloring agent, correctives and sweetener, Injectable composition can be aqueous isotonic solutions or suspension, and suppository can be by lipomul or suspension preparation.These compositionss can be for aseptic and/or contain adjuvant, such as antiseptic, stabilizing agent, wetting agent or emulsifying agent, dissolution accelerator, be used for regulating salt and/or the buffer agent of osmotic pressure.In addition, they also contain the upper valuable material of other treatment.The suitable preparation that is used for the transdermal application comprises the compounds of this invention and the carrier of effective dose, carrier can comprise that absorbable pharmaceutically acceptable solvent is to help Host Skin, for example, transdermal device is form of bandage, comprise the parts of mounting (backing member), contain the reservoir of the chemical compound that optionally is mixed with carrier, optional rate controlled barrier with controlled and set rate chemical compound be delivered to Host Skin in time limit time expand again and make device the apparatus of fixing with skin.Can also use the substrate preparation capable of permeating skin.Be used for the part, the suitable preparation that for example is applied in skin and eye is preferably aqueous solvent well-known in the art, ointment, cream or gel.This class preparation can contain solubilizing agent, stabilizing agent, tension-elevating agent, buffer agent and antiseptic.
Chemical compound of the present invention can be co-administered (drug regimen) with treatment effective dose and one or more treatments.For example according to pathology to be treated is arranged, be combined with other material that is used for the treatment of cardiovascular, diabetes, inflammation and/or neurodegenerative disease synergism can occur.The example of this compounds comprises medicine such as metformin, sulfonylurea, acarbose, peroxisome proliferator receptor PPAR γ activator, insulin, the special class (fibrates) of shellfish, his class D (statins) or other active substance glucagon-like (GLP-1), dipeptidase IV (DPP-IV), PPAR α/γ, PPAR δ/γ, PPAR δ and pan PPAR inhibitor, the 11 β hydroxyl steroid class dehydrogenases (inhibitor of 11 β-HSD), PTP 1B (PTP-1B) inhibitor, cannabis receptor (CB1) antagonist, glucagon receptor antagonist, pyruvate dehydrogenase kinase (PDK) inhibitor, hepatic glucose kinase activation agent etc.If chemical compound of the present invention and other therapies are given jointly, the dosage of the chemical compound that so jointly gives is certainly with the type of the medicine of common use, used concrete medicine, the disease for the treatment of etc. and different.
The present invention also provides drug regimen, and for example medicine box comprises 1) be the first activating agent of the compounds of this invention as herein described, it is free form or pharmaceutically acceptable salt form; 2) activating agent of at least a common use (co-agent).This medicine box comprises its administration description.
Term used herein " co-administered " or " administering drug combinations " refer to and comprise and give selected therapeutic agent and in order to comprise that activating agent wherein is not necessarily by identical route of administration or the therapeutic scheme that gives simultaneously to single patient.
Term as herein described " drug regimen " refers to and mixes or merge more than one active components and produce and comprise the medical product fixing and combination of on-fixed active component.Term " fixed combination " refers to single entities or dosage and gives active component to the patient simultaneously, suc as formula the chemical compound among the I-III and the common activating agent that uses.When term " on-fixed combination " refers to active component, for example formula I-III chemical compound and the common activating agent that uses as corpus separatum, follow or do not have concrete time showing successively the patient to be given, wherein this class administration provides 2 kinds of chemical compounds of effect level in patient body, the latter also can be applicable to HAART, for example gives active component more than 3 kinds or 3 kinds.
The method for preparing the compounds of this invention
Prepare the method for formula I-III chemical compound of the present invention and physicochemical property thereof referring to described in the Publication about Document, its Chinese style I compound name is called Hopeaphenol; Formula II compound name is called two dehydrogenation vitisin As (7 '-8 '-didehydro-vitisinA); The formula III compound name be called vitisin A (vitisinA) (the Hopeaphenol document:, J.C.S., 1965,406; Tet.Lett., 1965,2713, Chem.Pharm.Bull., 1998,46,663-668. vitisin A document: Phytochemistry.1996,42 (4): 1163-1165; J Agric Food Chem.2006,13; 54 (25): 9559-64.; Phytochemistry.2000,53 (8): 1015-9; Acla PharmaeeutieaSiniea 2001,36 (12): 944-950.7 '-8 '-two dehydrogenations-vitisin A document: J.O.C1993,58:805; Tetrohedron 1998,54:6651-6660; Tetrohedron 1999,55:2529-2544).
Definition
Term with regard to lxr receptor " adjusting " refers to the biological activity (for example transcriptional regulatory of target gene) that activates lxr receptor and be correlated with in the LXR approach.The adjusting of lxr receptor can be up regulation (i.e. excitement, activation or stimulation) or down-regulation (being antagonism, inhibition or retardance).The model of action of LXR regulator can be for directly, for example as part by being combined with lxr receptor.Regulating also can be for indirectly, for example by in conjunction with and/or modify another kind of molecule, come combination and activation lxr receptor.
" treatment " refers to the method that alleviates or alleviate disease and/or its simultaneous phenomenon.
Description of drawings
The EC50 amount effect curve of positive control T0901317 under Fig. 1, the different incubation time
The EC50 amount effect curve of the T0901317 of the SRC-1 synthetic peptide system of Fig. 2, variable concentrations
The EC50 curve comparison diagram of Fig. 3, positive control medicine and compound H opeaphenol
Fig. 4, Macrophage derived foam cell oil red dyeing photo
200 * and 400 * be amplification
The impact that Fig. 5, Western blot (Western blot) test sample are expressed THP-1 Macrophage derived foamy cell ABCA1.Fig. 5-1,2 is respectively sample treatment 12h, 24h result.Control is blank (not dosing) group,
T10uM is 10uM positive control T0901317 processed group, T10uM+ATRA is 10uM positive control T0901317 and 10uM retinoic acid receptor X (RXR) agonist all-trans-retinoic acid (ATRA) co-treatment group, H10uM+ATRA is Hopeaphenol 10uM and ATRA 10uM co-treatment group, H30uM, H10uM, H3uM is respectively Hopeaphenol30uM, 10uM, 3uM processed group.
The specific embodiment
The description of preferred embodiment
Embodiment hereinafter and embodiment are described the activity of chemical compound in detail, further specify the present invention but do not play the restriction effect by embodiment hereinafter, these embodiment and embodiment only are used for task of explanation, this area staff can expect carrying out various modifications or change according to it, and these modifications or change are also included within the application's the essence and scope and claims scope.All open source literatures, patent and the patent application of this paper citation are incorporated herein by reference for all purposes.
Transcribe test and be used for estimating the ability that the compounds of this invention is regulated the LXR transcriptional activity.In brief, by transient transfection the expression vector of chimeric protein is imported mammalian cell with the lower Reporter gene vector cotransfection of adjusting Protein G AL4 binding site control that luciferase gene wherein is in yeast, described chimeric protein contains is combined territory (DBD) with the DNA of the Gal4 of ligand binding domain (LBD) fusion of people LXRa or LXR β.Under LXR regulator existence condition, the LXR transcriptional activity changes, and this can monitor by the change of luciferase level.If the cell of transfection is exposed to lxr agonist, the LXR-dependent transcription is active so increases and the luciferase level rising.
Front 48 hours of this experiment beginning is with HEK293 cell (1 * 10
7) be inoculated in 175cm
2In the culture bottle, culture medium is 10%FBS, the DMEM culture medium of 1% penicillin/streptomycin.By using the PBS washed cell, trypsin digestion and cell, the PBS cell counting also is adjusted to 100000 cells/ml culture with cell concentration.It is 1: 10 that every 500ul transfection mixture uses Gal4-LBD expression plasmid and Gal4 UAS-luciferase reporter gene plasmid ratio, the Fugene amount of reagent: the plasmid amount be 5: 2 (ul: ug) and the preparation of the culture medium of serum-free, and with this transfection mixture incubated at room 30 minutes.Cell is mixed (10mL cell culture/500ul transfection mixture) and incubation 30 minutes more at room temperature with transfection mixture, then changes cell 100ul/ hole over to 96 porocyte culture plates, with cell at 37 ℃ and 5.0%CO
2Lower continuation incubation 24 hours, to the 12-point doubling dilution liquid of every kind of test compounds (comprise formula I-III chemical compound and with reference to positive control chemical compound T0901317) preparation in DMSO, wherein the initial concentration of chemical compound is 5mM.Test compounds (1ul) is joined in each cell hole of Tissue Culture Plate, the blank hole adds with volume DMSO, with cell at 37 ℃ and 5.0%CO
2Lower again incubation 24 hours is with cell pyrolysis liquid/luciferase test buffer B right-Glo
TM(25ul, Promega) joins in each hole, and room temperature incubation 5 minutes changes the 96 holes solid base plate of white over to and measures uciferase activity, i.e. luminous number of photons.
By divided by the blank value that is present in the DMSO on each flat board, to original marking of luminous value, obtain inducing multiple value.Use Origin Software on Drawing S type single-point dosage effect equation, use Sigmoidalfit match dose effect curve, EC50 is defined as the concentration that chemical compound causes half response between maximum and the minima.Compare with the maximum that positive control lxr agonist T0901317 obtains by the response that chemical compound is caused, calculate relative potency (or percentages of efficacy)
With reference to positive control chemical compound T0901317 (known lxr agonist, CAS RN:293754-55-9.FEBS Letters.2003,536:6-11)
Show that according to this measurement result chemical compound of the present invention transcribes LXR and have activation, has up regulation LXR effect, Hopeaphenol, vitisin A, two dehydrogenations-vitisin A is respectively 3.03uM, 6.92uM and 10.23uM., the positive control chemical compound is 0.123uM, percentages of efficacy is respectively T0901317 peaked 9%, 7.2%, 8.61%, and (activity is the meansigma methods of active result more than 5 times.Show value is X ± S.D.)
By the compounds of this invention the mensuration of the biological nature of LXR has been proved them in potential importance and practicality that the disease that is caused by the lxr receptor dysfunction take treatment or prevention is used as active substance in the medical product of purpose, described disease is particularly hypercholesterolemia, dyslipidemia and obesity, diabetes, cardiovascular disease, specific neurodegenerative diseases and inflammation.
Embodiment 2, LXR-FP (fluorescence polarization)-activator is raised experiment altogether
The FP experiment is used for estimating the ability (for example receptor coactivator) that the compounds of this invention is directly raised in conjunction with the protein of LXR ligand binding domains (LBD) and promotion reinforcement LXR transcriptional activity.This not celliferous experiment in vitro uses and forms recombination fusion protein by LXR-LBD and purification labelling (such as His-tag), and the peptide of synthetic fluorescein isothiocyanate (FITC) labelling, this peptide derives from the nuclear receptor interaction of receptor coactivator albumen such as steroid receptor receptor coactivator 1 (SRC-1) in conjunction with the territory.In the presence of lxr agonist, fluorescent labeling receptor coactivator peptide is raised to LXR-LBD, thereby low fluorescence polarization value becomes high fluorescence polarization value, and the size variation by the polarized fluorescence value provides intensity that the receptor coactivator peptide raises and thus in conjunction with the indication of the intensity of LXR-LBD agonist.
Be aminoacid sequence 162-447 (GenBank accession NM_005693) for the aminoacid sequence 202-461 (GenBank accession NM_007121) of LXR α-LBD and LXR β clones in coli expression carrier PET32a (Merck) and expresses with fusion rotein, and with His-tag antibody affinity column purification.FITC fluorescent labeling peptide has the feature of LLXXLL, and aminoacid sequence is FITC-ILRKLLQE-OH.
FP method list of references (GENES ﹠amp; DEVELOPMENT2000,14:2831-2838) method, final FP reaction system is the 100ul system: 100nM FITC-labelling peptide, (initial concentration is 10mM for 400nm histag-LXR and mensuration chemical compound 1uL, to measure 12 gradients of chemical compound doubling dilution with DMSO), reaction buffer consists of 10mM Hepes, 150mM NaCl, 2mM MgCl
2, 5mM, DTT at pH 7.9 still adopts T0901317 as positive control, and the DMSO hole is blank.With the dull and stereotyped incubated at room in 100ul reactant mixture place, every hole 2 hours, read the FP value.
The EC50 amount effect curve of positive control T0901317 under the different incubation times; The EC50 amount effect curve of the T0901317 of the SRC-1 synthetic peptide system of variable concentrations; The EC50 amount effect curve comparison diagram of positive control medicine and compound H opeaphenol is seen accompanying drawing 1-3
Be worth as a setting with the DMSO value, use Origin Software on Drawing S type single-point dosage effect equation, use Sigmoidal fit match dosage response curve, EC50 is defined as the concentration that chemical compound causes half response between maximum and the minima.Compare with the maximum that positive control lxr agonist T0901317 obtains by the response that chemical compound is caused, calculate relative potency (or percentages of efficacy).
Chemical compound of the present invention shows valuable pharmacological characteristics, such as above-mentioned external LXR-FP (fluorescence polarization)-activator among the application raise altogether the experiment confirmation, it is that 0.289uM.Hopeaphenol is 4.379uM to transcribing the EC50T0901317 that raises with the direct combination of LXR α and promotion, vitisin A is 5.67uM, and two dehydrogenations-vitisin A is 7.65uM.
Embodiment 3, resveratrol oligomer suppress macrophage foam cell formation and turn into experiment
Chemical compound of the present invention has been carried out the foamed inhibitory action research of macrophage.
The macrophage of cultivating is carried out trypsinization, and counting should be about 3 * 10 for cell density
5Individual/ml, be added on 24 well culture plates by the amount of every hole 1ml.Cell is divided into blank group, foam cell group and application of sample group.The blank group only adds serum-free medium, it is 50 μ g/ml to every hole final concentration that the foam cell group adds ox-LDL in addition, and the application of sample group adds testing sample again on the basis of foam group, put into 37 ℃ behind the mixing and contain 5%CO, cultivate oil red 0 dyeing of carrying out to dye neutral lipid after 48 hours in the incubator.
Through hatching of the macrophage processed with OX-LDL in 48 hours, the result does not add the OX-LDL model group cell of medicine as shown in Figure 1, and cellular morphology is imperfect, is foamed, and the oil droplet of a lot of oil red dyeing is arranged, and is typical Macrophage derived foam cell.Cellular morphology through compound treatment is more complete, and the dyeing of relative comparison cell oil red obviously shoals.This explanation chemical compound has inhibitory action to the foamed of macrophage.Meet the lxr agonist mechanism of action.Experimental result is seen accompanying drawing 4.
The short macrophage cholesterol efflux experiment of embodiment 4 resveratrol oligomer
THP-1 macrophage 0.2mCiL
-1[
3H] cholesterol and 50mg/L ox-LDL and contain 10% calf serum RPMI1640 culture fluid and jointly hatch after the 48h, with PBS liquid washed cell, again with containing 2gL
-1Bovine serum albumin RPMI1640 training liquid cultured cell 24h, and in culture fluid, add various influence factors, matched group adopts the blank RPMI1640 of adding culture medium.Afterwards, use again PBS liquid washed cell, containing 10mgL
-1 Cultured cell 12h in the pure and fresh culture fluid of apoA-I depletion of blood, scintillation counting technique detect respectively in culture fluid and the cell [
3H] cholesterol, Cholesterol Efflux, multiply by 100% again and represents divided by total CPM with CPM in the culture fluid.By detecting the impact on the THP-1 Macrophage derived foamy cell, result such as table 2, the demonstration chemical compound shows by activation LXR the Cholesterol Efflux that is mediated by apoA-1 in the THP-1 Macrophage derived foamy cell is the concentration dependent increase, meets lxr agonist active function mechanism.The result is as shown in table 1.
Table 2:
| Cell is processed | Cholesterol efflux |
| Without ox-LDL | 8.993±2.460 |
| Without ApoAI | 9.824±0.350 |
| Blank+ox-LDL 50ug/ml+ApoAI | 10.021±0.764 |
| T10uM | 23.069±4.504 ** |
| Two dehydrogenations vitisin A-30uM+ox-LDL 50ug/ml | 15.944±2.049 ** |
| Two dehydrogenations vitisin A-10uM+ox-LDL 50ug/ml | 14.085±0.518 ** |
| Two dehydrogenations vitisin A-3uM+ox-LDL 50ug/ml | 13.882±3.314 |
| Vitisin A+ox-LDL 50ug/ml | 13.949±0.505 ** |
| Vitisin A+ox-LDL 50ug/ml | 13.686±2.101 * |
| Vitisin A+ox-LDL 50ug/ml | 13.109±2.102 |
| Hopeaphenol 30uM+ox-LDL 50ug/ml | 16.788±0.950 ** |
| Hopeaphenol 10uM+ox-LDL 50ug/ml | 15.551±1.070 ** |
| Hopeaphenol 1uM+ox-LDL 50ug/ml | 14.766±0.210 ** |
Each sample activity is the meansigma methods of active result more than 3 times.Show value is the positive contrast of X ± S.D.T T0901317, and oxLDL is OxLDL ELISA
Embodiment 5 resveratrol oligomer are to the gene expression RT-PCR experiment of LXRa regulation and control target gene ABCA1 and apolipoprotein E
THP-1 macrophage (1 * 10
7Individual cell/bottle) in containing (or not containing) 50mg/L ox-LDL culture fluid, cultivates 48h, then add medicine cultured cell 24h (or 12h) with serum-free medium, matched group adopts the blank RPMI1640 of adding culture medium, PBS liquid washed cell, continue cultured cell 12h with 10mg/L apoA-I, collect and respectively organize cell.Extract total RNA by TRIzol test kit description.Get and respectively organize the synthetic cDNA of cell total rna 2 μ g reverse transcriptions, get again the reverse transcription product and adopt TakaraSYBR Premix Ex Taq
TMThe real-time quantitative PCR test kit carries out real-time quantitative PCR, asks for the Ct value, the calculation expression relative variation. and the result shows that medicine all can increase the mrna expression of ABCA1, ApoE, and is obvious dose dependent, meets the lxr agonist mechanism of action.
The ABCA1 amplification primers adopts GenebankNM_005502, forward primer ATTCGCTCTGAGATGAGCACCA; Reverse primer TTTCAAGCGGGCATAGAACCA, amplified production length 114bp.
The ApoE amplification primers adopts GenebankNM_000041, forward primer GCTGCGTTGCTGGTCACAT; Reverse primer TGCACCCAGCGCAGGTAAT, amplified production length is 161bp
Experimental result sees Table 2.
Table 2:
| Detect gene (different pharmaceutical processed group) | Ct value (12h) | Ct value (24h) | Relative variation (12h) | Relative variation (24h) |
| GAPDH is blank | 15.58±0.04 | 15.20±0.25 | ||
| GAPDH T10uM | 15.40±0.09 | 16.06±0.29 | ||
| GAPDH T090131710uM+ATRA10uM | 15.13±0.23 | 16.16±1.07 | ||
| GAPDH Hopeaphenol 10uM+ATRA10uM | 15.92±0.35 | 15.33±0.23 | ||
| GAPDH Hopeaphenol 30uM | 15.68±0.44 | 15.37±0.03 | ||
| GAPDH Hopeaphenol 10uM | 16.20±0.01 | 15.48±0.17 | ||
| GAPDH Hopeaphenol 3uM | 15.39±0.30 | |||
| ABCA1 is blank | 22.05±0.28 | 22.76±0.11 | ||
| ABCA1 T090131710uM | 20.31±0.08 | 22.35±0.26 | 3.19 | 2.41 |
| ABCA1 T090131710uM+ATRA10uM | 19.49±0.41 | 21.94±0.08 | 4.64 | 3.43 |
| ABCA1 Hopeaphenol 10uM+ATRA10uM | 19.32±0.33 | 19.81±0.50 | 9.06 | 8.46 |
| ABCA1 Hopeaphenol 30uM | 19.29±0.43 | 21.05±0..33 | 7.83 | 3.69 |
| ABCA1 Hopeaphenol 10uM | 20.17±0.07 | 21.26±0.03 | 6.10 | 3.42 |
| ABCA1 Hopeaphenol 3uM | 20.40±0.15 | 2.95 | ||
| GAPDH is blank | 15.05±0.28 | |||
| GAPDH T10uM | 15.11±0.35 | |||
| GAPDH T10uM+ATRA10uM | 15.60±0.84 | |||
| GAPDH Hopeaphenol 10uM+ATRA10uM | 15.49±0.002 | |||
| GAPDH Hopeaphenol 30uM | 15.16±0.003 | |||
| GAPDH Hopeaphenol 10uM | 15.11±0.347 | |||
| ApoE is blank | 22.26±0.27 | |||
| ApoE T10uM | 20.61±0.50 | 5.389 | ||
| ApoE T10uM+ATRA10uM | 20.01±0.27 | 7.062 | ||
| ApoE Hopeaphenol 10uM+ATRA10uM | 20.13±0.34 | 5.12 | ||
| ApoE Hopeaphenol 30uM | 20.35±0.50 | 4.76 | ||
| ApoE Hopeaphenol 10uM | 20.36±0.47 | 3.91 |
Each sample activity is the meansigma methods of active result more than 4 times.Show value is the positive contrast of X ± S.D.T T0901317, and ATRA is the rxr agonist all-trans-retinoic acid, and GAPDH is the reference gene contrast
Embodiment 6 resveratrol oligomer are to THP-1 Macrophage derived foamy cell ABCA1 protein expression Western blot
Macrophage derived foamy cell extracts total cellular protein and detects the ABCA1 protein expression through Western blot (Western blot) after drug treating.
After in the good cell of results, adding without the digestion of enzymic digestion liquid, carry out lysis, in 4 ℃ of centrifugal 10min, the reject precipitation, the BCA method is carried out protein quantification, gets 50 μ g albumen and adds in 4 * sds gel sample loading buffer, heats 10min so that protein denaturation at 100 ℃.Carry out electrophoretic separation with the 7%SDS-polyacrylamide gel, turn the PDVF film, adopt and dye in advance protein standard to determine the albumen position.Confining liquid sealing 4h, added goat-anti people ABCA1 primary antibodie by 1: 200,4 ℃ of overnight incubation, TBST washes 5 times, adding the anti-sheep two of horseradish peroxidase-labeled rabbit at 1: 2000 resists, incubated at room 1h, TBST wash 5 times, detect with Western blot trace fluoroscopic examination ECL test kit, the results are shown in accompanying drawing 2, the ABCA1 protein expression is compared obvious increase with the blank group after drug treating, and 12h and 24h all are fine dose-dependence, meets the mechanism of action of lxr agonist.
Embodiment 7 resveratrol oligomer suppress macrophage tumor necrosis factor (TNF α) and interleukin-1 beta (IL-1 β) secretion that lipopolysaccharide (LPS) is induced
Embodiment uses TNF α and its antiinflammatory action of IL-1 β experimentation.This experiment utilizes macrophage can secrete a large amount of inflammatory cytokines after derivant stimulates, and observes sample to the impact of TNF α and IL-1 β secretion level.In 24 orifice plate reaction systems, contain and have an appointment 2.0 * 10
5/ ml person monocytic cell, stimulant is induced differentiation 48h, makes into macrophage, add positive control medicine T0901317 and medicine to be measured, establish simultaneously negative control hole, replace testing sample with the RPMI1640 culture fluid, preincubate 30min adds lipopolysaccharide LPS, 37 ℃, 5%CO
2Hatch 24h, get supernatant 100ul, adopt enzyme linked immunological (ELISA) method to detect.
ELISA method operation sequence:
From test kit, take out lath, add standard substance and the testing sample of 100 μ l variable concentrations, hatch 90min for 37 ℃.Wash plate 3 times, add the biotinylated antibody working solution, 60min is hatched for 37 ℃ in 100 μ l/ holes.Wash plate 3 times, add enzyme in conjunction with working solution, 60min is hatched for 37 ℃ in 100 μ l/ holes.Wash plate 3 times, add developer, 100 μ l/ holes, 37 ℃ of lucifuges are hatched 20min.Add stop buffer, the OD450 value is measured in 100 μ l/ holes in the 5min behind the mixing.Take standard substance concentration as abscissa, take the OD450 value as vertical coordinate, draw IL-1 β concentration-OD450 value standard curve, then find the concentration of testing sample from standard curve according to the OD450 value of measured sample, and calculate sample with respect to the ratio of contrast.TNF α, IL-1 β inhibitory action that medicine is secreted macrophage the results are shown in Table 3-5 and show that chemical compound composition of the present invention has antiinflammatory action, meet LXR regulatory mechanism effect characteristics.
Table 3 resveratrol oligomer suppresses the person monocytic cell source property macrophage THP-1 TNF α secretion experimental result that LPS stimulates 24h
| The cell different pharmaceutical is processed | TNF α secretes (with respect to blank) |
| |
1 |
| LPS 5ug/ml | 1.525±0.029 ** |
| T10uM+LPS 5ug/ml | 1.284±0.024## |
| Two dehydrogenations vitisin A-30uM+LPS 5ug/ml | 1.287±0.053## |
| Two dehydrogenations vitisin A-10uM+LPS 5ug/ml | 1.423±0.011## |
| Two dehydrogenations vitisin A-3uM+LPS 5ug/mL | 1.433±0.040# |
| Two dehydrogenations amurensisin-1uM+LPS 5ug/ml | 1.50±0.038 |
| Vitisin A-30uM+LPS 5ug/ml | 1.269±0.021## |
| Vitisin A-10uM+LPS 5ug/ml | 1.370±0.047## |
| Vitisin A-3uM+LPS 5ug/ml | 1.472±0.049 |
| Vitisin A-1uM+LPS 5ug/ml | 1.517±0.047 |
| Hopeaphenol 30uM+LPS 5ug/ml | 1.262±0.020## |
| Hopeaphenol 10uM+LPS 5ug/ml | 1.289±0.025## |
| Hopeaphenol 3uM+LPS 5ug/ml | 1.368±0.026## |
| Hopeaphenol 1uM+LPS 5ug/ml | 1.471±0.071 |
Data (mean ± S.D., n=3);
*P＜0.01vs.control, #P＜0.05, the ##P＜positive contrast of 0.01vs.LPS treatmentonly.Student-Newman-Keuls test.T T0901317
Table 4LPS stimulates mouse macrophage RAW264.7 24h experimental result
| The cell different pharmaceutical is processed | IL-1 β secretes (with respect to blank) |
| |
1 |
| LPS 5ug/ml | 1.924±0.108 ** |
| T10uM+LPS 5ug/ml | 1.361±0.169## |
| Two dehydrogenations vitisin A-30uM+LPS 5ug/ml | 1.013±0.237## |
| Two dehydrogenations vitisin A-10uM+LPS 5ug/ml | 1.370±0.160## |
| Two dehydrogenations vitisin A-3uM+LPS 5ug/mL | 1.887±0.127 |
| Vitisin A-30uM+LPS 5ug/ml | 1.183±0.136## |
| Vitisin A 10uM+LPS 5ug/ml | 1.462±0.098## |
| Vitisin A-3uM+LPS 5ug/ml | 1.687±0.069# |
| Hopeaphenol 30uM+LPS 5ug/ml | 1.035±0.296## |
| Hopeaphenol 10uM+LPS 5ug/ml | 1.457±0.136## |
| Hopeaphenol 3uM+LPS 5ug/ml | 1.768±0.061 |
Data (mean ± S.D., n=3);
*The blank group of P＜0.01vs., #P＜0.05, the ##P＜positive contrast of 0.01vs.LPS processed group only.Student-Newman-Keuls test.T T0901317
Table 5.LPS stimulates mouse macrophage RAW264.7 24h experimental result
| The cell different pharmaceutical is processed | TNF α secretes (with respect to blank) |
| |
1 |
| LPS 5ug/ml | 1.711±0.057 ** |
| T10uM+LPS 5ug/ml | 1.449±0.062## |
| Two dehydrogenations vitisin A-30uM+LPS 5ug/ml | 1.047±0.156## |
| Two dehydrogenations vitisin A-10uM+LPS 5ug/ml | 1.277±0.055## |
| Two dehydrogenations vitisin A-3uM+LPS 5ug/mL | 1.447±0.098# |
| Vitisin A-30uM+LPS 5ug/ml | 1.259±0.095## |
| Vitisin A-10uM+LPS 5ug/ml | 1.499±0.050## |
| Vitisin A-3uM+LPS 5ug/ml | 1.652±0.071 |
| Hopeaphenol 30uM+LPS 5ug/ml | 1.352±0.035## |
| Hopeaphenol 10uM+LPS 5ug/ml | 1.512±0.064# |
| Hopeaphenol 3uM+LPS 5ug/ml | 1.529±0.122 |
Data (mean ± S.D., n=3);
*The blank group of P＜0.01vs., #P＜0.05, the ##P＜positive contrast of 0.01vs.LPS processed group .Student-Newman-Keuls test.T T0901317
Embodiment 8 resveratrol oligomer produce the inhibitory action that nitric oxide produces to macrophage
Experimental procedure
The THP-1 cell is with containing 10% calf serum RPMI1640 culture fluid, leaves standstill cultivation in 37 ℃, 5%CO2 incubator.Hatch THP-1 mononuclear cell 48h with 160nM PMA before each experiment, it is induced be divided into macrophage.With cell according to 2 * 10
4Cell number add 96 orifice plates, treat that cell attachment is thick, macrophage is divided into LPS group (LPS final concentration 5 μ g/ml), the LPS+ sample sets, 24h is processed in each 4 multiple holes of each group, gets cells and supernatant, for detection of the concentration of NO.Get respectively and respectively organize cells and supernatant 100 μ l, add Griess reagent (1% p-anilinesulfonic acid., 0.1%N-naphthyl diethylamine, 2.5% phosphoric acid), build up chemical method NO testing cassete operating instruction according to Nanjing, in 550nm place colorimetric, measure its optical density with microplate reader.The 6 explanation the compounds of this invention that the results are shown in Table of this research have stronger inhibitory action to the generation of macrophage NO, and good dose-dependence is arranged, and meet the mechanism of action of lxr agonist.
Table 6 resveratrol oligomer suppresses LPS and induces THP-1 cell NO to discharge the result
| THP-1 cell different pharmaceutical is processed | Content of nitric oxide (uM) |
| Blank | 3.57±2.75 |
| LPS 5ug/ml | 45.72±5.39 ** |
| T10uM+LPS 5ug/ml | 12.53±5.46## |
| Two dehydrogenations vitisin A-30uM+LPS 5ug/ml | 4.28±2.58## |
| Two dehydrogenations vitisin A-10uM+LPS 5ug/ml | 7.63±4.64## |
| Two dehydrogenations vitisin A-3uM+LPS 5ug/ml | 29.81±5.96## |
| Vitisin A-30uM+LPS 5ug/ml | 4.81±3.19## |
| Vitisin A-10uM+LPS 5ug/ml | 22.03±5.21## |
| Vitisin A-3uM+LPS 5ug/ml | 34.42±6.93# |
| Hopeaphenol 30uM+LPS 5ug/ml | 12.01±4.97## |
| Hopeaphenol 10uM+LPS 5ug/ml | 23.23±6.47## |
| Hopeaphenol 1uM+LPS 5ug/ml | 35.40±5.80# |
Data (mean ± S.D., n=4);
*P＜0.01vs. blank, #P＜0.05, the ##P＜positive contrast of 0.01vs.LPS processed group .Student-Newman-Keuls test.T T0901317
Claims (2)
1. the resveratrol tetramer compound, its Hopeaphenol shown in the formula (I) uses in the medicine of preparation prevention and treatment hypercholesterolemia
2. suc as formula (II) resveratrol tetramer compound, its 7 " '-8 " '-two dehydrogenation vitisin A (7 " '-8 " '-didehydro-vitisinA) and suc as formula (III) vitisin A (vitisinA) in the medicine of preparation prevention and treatment hypercholesterolemia, use
Formula (II) formula (III).
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| CN105147660A (en) * | 2015-08-13 | 2015-12-16 | 中国科学院西北高原生物研究所 | Application of resveratrol oligomer in drug preparation |
| CN109662962B (en) * | 2018-10-26 | 2022-02-11 | 中国科学院西北高原生物研究所 | Use of oligomeric stilbenes |
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| CN110511200B (en) * | 2019-09-23 | 2023-05-16 | 中国科学院西北高原生物研究所 | Method for separating Hopephenol or/and Isohopyenol from Iris lactea seed |
| CN114869876B (en) * | 2022-06-10 | 2024-09-06 | 中国科学院西北高原生物研究所 | Application of oligomeric stilbene compounds in preparation of anti-inflammatory products |
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