CN105582005A - Novel medical application of isopentenyl isoflavone compounds in licorice roots - Google Patents

Novel medical application of isopentenyl isoflavone compounds in licorice roots Download PDF

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CN105582005A
CN105582005A CN201510918824.1A CN201510918824A CN105582005A CN 105582005 A CN105582005 A CN 105582005A CN 201510918824 A CN201510918824 A CN 201510918824A CN 105582005 A CN105582005 A CN 105582005A
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licoricidin
compounds
inhibition
ptp1b
activity
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CN105582005B (en
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叶敏
余四旺
季帅
李紫薇
王永瑞
唐叔南
乔雪
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Peking University
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    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/33—Heterocyclic compounds
    • A61K31/335—Heterocyclic compounds having oxygen as the only ring hetero atom, e.g. fungichromin
    • A61K31/35—Heterocyclic compounds having oxygen as the only ring hetero atom, e.g. fungichromin having six-membered rings with one oxygen as the only ring hetero atom
    • A61K31/352—Heterocyclic compounds having oxygen as the only ring hetero atom, e.g. fungichromin having six-membered rings with one oxygen as the only ring hetero atom condensed with carbocyclic rings, e.g. methantheline 
    • A61K31/353—3,4-Dihydrobenzopyrans, e.g. chroman, catechin
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/33—Heterocyclic compounds
    • A61K31/335—Heterocyclic compounds having oxygen as the only ring hetero atom, e.g. fungichromin
    • A61K31/35—Heterocyclic compounds having oxygen as the only ring hetero atom, e.g. fungichromin having six-membered rings with one oxygen as the only ring hetero atom
    • A61K31/352—Heterocyclic compounds having oxygen as the only ring hetero atom, e.g. fungichromin having six-membered rings with one oxygen as the only ring hetero atom condensed with carbocyclic rings, e.g. methantheline 
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K8/00—Cosmetics or similar toiletry preparations
    • A61K8/18—Cosmetics or similar toiletry preparations characterised by the composition
    • A61K8/30—Cosmetics or similar toiletry preparations characterised by the composition containing organic compounds
    • A61K8/49—Cosmetics or similar toiletry preparations characterised by the composition containing organic compounds containing heterocyclic compounds
    • A61K8/4973—Cosmetics or similar toiletry preparations characterised by the composition containing organic compounds containing heterocyclic compounds with oxygen as the only hetero atom
    • A61K8/498—Cosmetics or similar toiletry preparations characterised by the composition containing organic compounds containing heterocyclic compounds with oxygen as the only hetero atom having 6-membered rings or their condensed derivatives, e.g. coumarin
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61Q—SPECIFIC USE OF COSMETICS OR SIMILAR TOILETRY PREPARATIONS
    • A61Q19/00—Preparations for care of the skin
    • A61Q19/02—Preparations for care of the skin for chemically bleaching or whitening the skin

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Abstract

The invention discloses novel medical application of isopentenyl isoflavone compounds in licorice roots. Through systematic research on the chemical components and activity screening of the licorice roots, it is discovered that the isopentenyl isoflavone compounds including licoricidin, angustone A and glyasperin D of which the structures are similar have the significant activity on inhibition of cancer cell proliferation and inhibition of protein tyrosine phosphatase 1B (PTP1B), licoricidin and glyasperin D have the significant activity on inhibition of acetylcholine esterase (AChE), and glyasperin D also has the significant activity on inhibition of tyrosinase. On the basis of the discovery, the active compounds or salt, ester and solvent compounds, stereoisomers, tautomer and prodrugs which are acceptable in pharmacy of the active compounds and mixtures of the compounds can be used for preparing anti-cancer drugs, PTP1B inhibitors, antidiabetic drugs, tyrosinase inhibitors, freckle removing and whitening products, AChE inhibitors, anticholinesterase drugs and the like.

Description

The new medical use of an isoprenoid isoflavones compound in Radix Glycyrrhizae
Technical field
The present invention relates to the new medicine use of an isoprenoid isoflavones compound in Radix Glycyrrhizae, be specifically related to theyPrepare in cancer therapy drug, PTP1B inhibitor and antidiabetic medicine, tyrosinase inhibitor, acetylcholinesteraseinhibitors inhibitorsApplication.
Background technology
Radix Glycyrrhizae (GlycyrrhizauralensisFisch.) is one of the most frequently used Chinese medicine, has had 2000 in ChinaApplicating history for many years, except Chinese pharmacopoeia, it is also included by Japanese Pharmacopoeia, European Pharmacopoeia and American Pharmacopeia. Radix Glycyrrhizae toolHave clearing heat and detoxicating, expelling phlegm and arresting coughing, coordinating the drug actions of a prescription, the multiple efficacies such as invigorate the spleen and benefit qi, be mainly used in weakness of the spleen and the stomach, coughing with a lot of sputum, the heartThrob with fear breathe hard, the illness such as carbuncle sore tumefacting virus, in modern clinic application, be mainly used in treating respiratory tract infection, gastric ulcer, diabetes withAnd the disease such as virus hepatitis. In addition, Radix Glycyrrhizae is also widely used in cosmetic industry, has significant freckle removing and whitening equivalenceReally. The chemical composition of Radix Glycyrrhizae is numerous, from glycyrrhiza genus, has separated so far approximately 450 compounds of report, is mainly divided into threeTerpene saponins, flavonoid glycoside metaclass and flavone glycoside compounds. Wherein, the aglycon of triterpenoid saponin is mainly oleanane type threeTerpene, and flavone aglycone compounds kind is a lot, mainly comprise flavones, flavonols, flavanone, isoflavones, isoflavanone,Isoflavan, red sandalwood alkane, chalcone and cumarin etc. Mainly concentrate for the pharmacology activity research of chemical composition in Radix Glycyrrhizae at presentThe higher compound of some content in Radix Glycyrrhizae, such as glycyrrhizic acid, glycyrrhizin, isoliquiritigenin, liquiritin and isoliquiritinDeng, and seldom have report for the pharmacology activity research of other compositions.
Licoricidin (English name: licoricidin; No. CAS: 30508-27-1; Molecular formula: C26H32O5; Molecular weight:424), AngustoneA (No. CAS: 90686-13-8; Molecular formula: C25H26O6; 422) and Glycyrrhiza Aspera Root element D (English molecular weight:Name: glyasperinD; No. CAS: 142561-10-2; Molecular formula: C22H26O5; Molecular weight: 370) be the chemical composition of Radix Glycyrrhizae,Their chemical constitution is similar, all belongs to isoflavones or the isoflavan compounds of isopentene group, at its C-5, C-7, C-2'With on C-4', all have hydroxyl or methoxy substitution, on C-6, have isopentene group replace, in addition, at licoricidin and angustoneOn the C-3' of A, there is second isopentene group to replace.
Licoricidin be once in the news and there is the cAMP phosphodiesterase of inhibition (KusanoA etc.,Chem.Pharm.Bull.1991,39,930-933), suppress haemolysis platelet activating factor transacetylase (NagumoS etc.,Biol.Pharm.Bull.1999,22,1144-1146), antibacterial (HatanoT etc., Chem.Pharm.Bull.2000,48,1286-1292; TanakaY etc., J.Nutr.Sci.Vitaminol.2001,47,270-273; FukaiT etc., LifeSci.2002,71,1449-1463; TanabeS etc., JBreathRes.2012,6,016006; VillinskiJR etc., JNatProd.2014,77,521-526; Eerdunbayaer etc., Molecules2014,19,13027-13041;KirmizibekmezH etc., Fitoterapia2015,103,289-293), anti-ephritis and oxidation (FukaiT etc.,Fitoterapia2003,74,720-724), the mouth disease such as treatment periodontitis (LaVD etc., JPeriodontol.2011,82,122-128; MessierC etc., OralDis.2012,18,32-39; GafnerS etc., JNatProd.2011,74,2514-2519) isoreactivity, also once had and reported that the licorice that contains licoricidin can suppress prostate gland cancer cellThe transfer ability (ParkSY etc., BrJNutr.2010,104,1272-1282) of DU145, but have no straight about licoricidinConnect the report of inhibition tumor cell propagation and PTP1B activity.
There is not yet any report about the activity of AngustoneA, and hydroxyl on isopentene group and C-7 on its C-6Dehydrocyclization and the derivative AngustoneC that generates has antibacterial and pipe intestinal protection (QuesadaL etc., NatProdCommun.2012,7,1187-1188) effect.
Glycyrrhiza Aspera Root element D was once in the news and had weak anti-helicobactor pylori activity (FukaiT etc., LifeSci.2002,71,1449-1463), have no in addition other active report.
PTP 1B (PTP1B) is single-minded hydrolysis aromatic phosphoric acid in vivo, by insulin receptor orTyrosine residue dephosphorylation on its substrate, to insulin signaling, negative regulator is carried out in transduction, and PTP1B inhibitor can be used forTreatment type ii diabetes. Research in recent years shows that PTP1B has also participated in cancer and developed, and its inhibitor is at anticancer aspectThere is great potential (LessardL etc., BiochimBiophysActa.2010,1804,613-619). Acetylcholinesterase(AChE) be optionally hydrolyzed in vivo acetylcholine and generate choline and acetic acid, reduce the physiological effect of cholinergic nerve, AChEInhibitor can be used for treating nervous system degenerative disease as alzheimer disease etc. Tyrosinase is that in skin, melanin is syntheticKey enzyme, it is melanic synthetic in body that tyrosinase inhibitor can suppress, and has the effect of freckle removing and whitening. These target spots andThe pharmacological action of Radix Glycyrrhizae is closely related, also has and reports that some derive from Radix Glycyrrhizae and compound or extract and can suppress these targetsPoint, but the relevant activity of three kinds of compounds disclosed in this invention is not reported.
Summary of the invention
The object of this invention is to provide in Radix Glycyrrhizae three kinds of prenyl isoflavones compounds (be licoricidin,AngustoneA and Glycyrrhiza Aspera Root element D) new medicine use. The structural formula of these three kinds of compounds is as follows:
The new medicine use of three kinds of compounds provided by the present invention refers to these compounds itself or it is pharmaceutically acceptableSalt, ester, solvate, stereoisomer, dynamic isomer, prodrug at least one, and their mixture is in systemApplication in the medicine of at least one function in following (1)-(5) of getting everything ready: (1) cancer therapy drug; (2) antidiabetic medicine; (3)PTP1B inhibitor; (4) acetylcholinesteraseinhibitors inhibitors; Or (5) tyrosinase inhibitor.
The inventor, by system separation and purification and various active screening verification to Radix Glycyrrhizae chemical composition, has found someThe individual compound with remarkable activity. The wherein disclosed three kinds of compounds of the present invention, i.e. licoricidin, angustoneA and thickHair glycyrrhizin D, is the close prenyl isoflavones compounds of structure, has outstanding inhibition tumor cell survival and suppressesThe activity of PTP1B and the activity of acetylcholine esterase inhibition, Glycyrrhiza Aspera Root element D also has the remarkable work of restraint of tyrosinaseProperty. Further research is found, licoricidin is the propagation of inhibition tumor cell effectively, the apoptosis of induced tumor cell.AngustoneA can suppress kinds of tumor cells vigor by cell death inducing and autophagy approach, its active and its inhibitionTumour cell glycolysis ability is relevant.
According to the present invention, licoricidin, angustoneA and Glycyrrhiza Aspera Root element D can be used for preparing cancer therapy drug or PTP1BInhibitor and antidiabetic medicine, licoricidin and Glycyrrhiza Aspera Root element D can be used for preparing acetylcholinesteraseinhibitors inhibitors, and coarse wool is sweetThe plain D of grass also can be used for preparing tyrosinase inhibitor and freckle removing and whitening product.
With above-mentioned reactive compound or its pharmaceutically acceptable salt, ester, solvate, stereoisomer, tautomerismBody, prodrug and their mixture are that medicine prepared by active ingredient also belongs to protection scope of the present invention.
When needs, in said medicine, can also add one or more pharmaceutically acceptable carrier or auxiliary materials.Described carrier or auxiliary material comprise diluent, excipient, filler, adhesive, wetting agent, disintegrant, the suction of pharmaceutical field routineReceive promoter, surfactant, absorption carrier, lubricant etc.
Utilize above-mentioned reactive compound or its pharmaceutically acceptable salt, ester, solvate, stereoisomer, change differentStructure body and prodrug, as active component, are used alone or in combination, or prepare into various doses with other medicines, auxiliary material etc.Type, includes but not limited to the various ways such as tablet, powder, pill, injection, capsule, film, suppository, paste, electuary. OnThe medicine of stating various formulations all can be according to the conventional method preparation of pharmaceutical field.
Brief description of the drawings
Fig. 1 be licoricidin proton nmr spectra (1HNMR)。
Fig. 2 be licoricidin carbon-13 nmr spectra (13CNMR)。
Fig. 3 be angustoneA proton nmr spectra (1HNMR)。
Fig. 4 be angustoneA carbon-13 nmr spectra (13CNMR)。
Fig. 5 be Glycyrrhiza Aspera Root element D proton nmr spectra (1HNMR)。
Fig. 6 be Glycyrrhiza Aspera Root element D carbon-13 nmr spectra (13CNMR)。
Fig. 7 is the effect of licoricidin to the SW480 human colon cancer cell cycle.
Fig. 8 is the effect of licoricidin to SW480 apoptosis of human colon cancer cells.
Fig. 9 is the effect of angustoneA to SW480 apoptosis of human colon cancer cells.
Figure 10 is that angustoneA suppresses the effect that SW480 human colon cancer cell clone forms.
Detailed description of the invention
The experimental technique using in following embodiment if no special instructions, is conventional method.
Material, reagent etc. used in following embodiment, if no special instructions, all can obtain from commercial channels.
The separation and purification of embodiment 1, licoricidin, angustoneA and Glycyrrhiza Aspera Root element D.
One, experiment material and method.
All chemical reagent of mentioning in this method are all purchased from Beijing Chemical Plant.
Get 35kg licorice medicinal materials (Radix Glycyrrhizae branch company of science and technology Industry Co.,Ltd of Inner Mongol Erie), pulverize, with 10 timesMeasure the each extraction twice of 95% ethanol and 70% ethanol, each 2～3 hours, decompression recycling ethanol, the medicinal extract obtaining (10L) is mixedBe suspended from water, be extracted with ethyl acetate 5 times, obtain ethyl acetate extract 2500g. Get 1280g ethyl acetate extract, through silicagel columnChromatogram (200～300 orders, Qingdao Marine Chemical Co., Ltd.) separates, petroleum ether-ethyl acetate gradient elution (1:0～1:1),Obtain flow point A～J. Flow point E separates through MCI column chromatography (the biological Co., Ltd of Chengdu science popularization), alcohol-water gradient elution (0:1～1:0), obtain flow point EA～EG. Flow point EC separates dichloro through polyamide column chromatography (the biochemical plastic molding and processing plant of Taizhou of Zhejiang road and bridge tetramethyl)Methane-methyl alcohol gradient is de-(1:0～1:1) first, obtains flow point ECA～ECN. Flow point ECC, ECD and ECH are respectively through ODSC18Post lookSpectrum (DAISO company, Japan) separate, methanol-water gradient elution (0:1～1:0), then through half preparative HPLC (Agilent company,The U.S.) purifying, obtain angustoneA70mg, Glycyrrhiza Aspera Root element D14mg, licoricidin 900mg.
Two, experimental result.
By the contrast of the nmr spectrum of three compounds that obtain and document, be accredited as respectively licoricidin (Yang Li etc.,Research and development of natural products. 2009,448,438-440), angustoneA (LaneGA etc.,Phytochemistry.1987,26,295-300) and Glycyrrhiza Aspera Root element D (ZengL etc., Heterocycles.1992,34,375-387), their proton nmr spectra (1HNMR) and carbon spectrum (13CNMR) as shown in Fig. 1～6.
Embodiment 2, licoricidin, angustoneA and Glycyrrhiza Aspera Root element D suppress multiple cancer cell vigor.
One, experiment material and method.
HepG2 human liver cancer cell, SW480 human colon cancer cell, A549 human lung carcinoma cell, MCF7 human breast cancer cell are all purchasedPreserve center (ATCC) from U.S.'s bacterial classification, all experiments all adopt the cell in exponential phase.
(1) cell was inoculated in 96 orifice plates after 12 hours, and adding respectively appointment Radix Glycyrrhizae compound to final concentration is 10 μ M, continuesCultivate 24 hours, then add MTS solution to final concentration to every hole be 0.5mg/mL, continue to cultivate 2～4 hours, then use fromMoving ELIASA is measured each hole absorbance in 490nm, calculates its inhibiting rate with respect to control group.
(2) cell was inoculated in 96 orifice plates after 12 hours, added respectively and specified Radix Glycyrrhizae compound to prescribed concentration, continued to cultivate24 hours, then add MTS solution to final concentration to every hole be 0.5mg/mL, continues to cultivate 2～4 hours, then uses automatic enzymeMark instrument is measured each hole absorbance in 490nm, calculates it according to each compound inhibiting rate with respect to control group under prescribed concentrationHalf-inhibition concentration (IC50) value.
Two, experimental result.
As shown in table 1, three kinds of compounds all can significantly suppress HepG2 human liver cancer cell, SW480 people's knot under 10 μ M concentrationThe vigor of colon-cancer cell, A549 human lung carcinoma cell, MCF7 human breast cancer cell. Further quantitative dose-effect relationship result shows,Except the IC of Glycyrrhiza Aspera Root element D to A549 cell50Value is outside 12.9 μ M, the IC that all the other all tests obtain50Value is all lower than 10 μM, the wherein IC of licoricidin to HepG2 cell50Value, even lower than 1 μ M, has reached 300nM. These results show, Radix Glycyrrhizae westFixed, angustoneA and Glycyrrhiza Aspera Root element D have the activity of the multiple cancer cell vigor of remarkable inhibition, can be used for preparing anticarcinogenThing.
Table 1, licoricidin, angustoneA and Glycyrrhiza Aspera Root element D suppress HepG2 human liver cancer cell, SW480 people's colonCancer cell, A549 human lung carcinoma cell, MCF7 human breast cancer cell vigor.
Embodiment 3, licoricidin and angustoneA suppress SW480 cell proliferation, induction SW480 Apoptosis.
One, experiment material and method.
(1) SW480 cell was inoculated in 6 orifice plates after 12 hours, added respectively licoricidin or angustoneA dense to specifyingDegree, continues to cultivate 24 hours, collects all floating and adherent cells, adds the ethanol of 1mL70%, and-20 DEG C are fixedly spent the night.By centrifugal the cell 200g fixing 5 minutes, supernatant discarded, added the PBS (Zhong Kemai Science and Technology Ltd. in morning) of 1mL, piping and drumingEvenly, again centrifugal. Then add the propidium iodide (PI) that contains 5 μ g/mL and RNase (the learned innovation in Beijing of 100 μ g/mLDevelopment in science and technology Co., Ltd), hatch 30 minutes for 37 DEG C, adopt FASCAN flow cytometer (BeckonDickenson company, U.S.State) to measure, excitation wavelength is 488nm, emission wavelength is 630nm.
(2) SW480 cell was inoculated in 6 orifice plates after 12 hours, added respectively licoricidin or angustoneA dense to specifyingDegree, continues to cultivate 6 hours, collects all floating and adherent cells, adds the PBS (Zhong Kemai Science and Technology Ltd. in morning) of 1mL,Piping and druming evenly, again centrifugal. Then add 400 μ LAnnexinV (Beijing Xin Shengke Science and Technology Ltd.) in conjunction with liquid by cellSuspendible, then add 5 μ LAnnexinV-FITC (Beijing Xin Shengke Science and Technology Ltd.), room temperature lucifuge was hatched after 15 minutes, thenAdd 10 μ L propidium iodides (PI, Beijing Xin Shengke Science and Technology Ltd.), lucifuge is hatched 10 minutes on ice. Adopt FASCAN streamingCell instrument (BeckonDickenson company, the U.S.) is measured red and green fluorescence situation.
Two, experimental result.
(1) reason that MTS method detection cell viability declines is mainly divided into two classes: cell proliferation minimizing or cell death increaseAdd. First, we have measured the work of licoricidin to the SW480 cycle by the method for flow cytometry propidium iodide (PI) dyeingWith. As shown in Figure 7, licoricidin can significantly block the fissional G1/S phase of SW480, and is dose dependent. At 10 μWhen M, substantially there is no the cell in G2 division stage. These data show that licoricidin can inhibition tumor cellPropagation. AngustoneA is mainly apoptosis-induced and do not affect cell cycle distribution.
(2) then, we adopt that a kind of conventional can to follow the trail of apoptotic method be the two dyeing of AnnexinV/PIMethod evaluate licoricidin to the apoptotic impact of SW480. As shown in Figure 8, the SW480 cell quilt of control groupAnnexinV and PI are high, and the ratio of dying is very low, processes after 6 hours the ratio of high transfect cell when giving the licoricidin of concentration gradientThe rising of example concentration dependent. Wherein, the high part of dying of AnnexinV has been indicated phosphatidyl silk on apoptosis starting stage cell membraneTurning up of propylhomoserin, and for two high parts of dying, the variation of indicator cells permeability, is commonly considered as the mark of apoptosis in late periodWill. These data show the apoptosis that licoricidin and angustoneA all can induced tumor cells.
Embodiment 4, angustoneA suppress SW480 cells in vitro clone and form.
One, experiment material and method.
It is 20～40/ml that the SW480 of exponential phase is diluted to concentration, the every hole inoculation of six orifice plates 2.5ml. Treat that cell pastesAfter wall, use respectively 0,0.1,0.5,1,2.5,5 μ MangustoneA to process after 48h, change normal complete medium into and continueContinuous while cultivating about the 10 days clonal expansions to Control group to about 50 cells, fixing and with after violet staining to oftenClone's number in hole is counted. The clone's number obtaining goes out angustoneA through the Fitting Calculation and suppresses the IC that clone forms50Value.
Two, experimental result.
Colony formation can reflect cell colony dependence and two important characters of multiplication capacity, is therefore often used asFor evaluating in vitro the goldstandard of cytotoxic activity of antineoplastic. We also adopt colony formation evaluationAngustoneA suppresses the effect of tumor growth.
Experimental result as shown in figure 10. By counting, obtaining angustoneA concentration is 0,0.1,0.5,1,2.5,5 μ MTime, clone's number is 97,78,52,32,12,0. By the IC obtaining after matching50Be 0.50 ± 0.06 μ M. The above results is aobviousShow that angustoneA has good antitumor activity and patent medicine prospect.
Embodiment 5, licoricidin, angustoneA and Glycyrrhiza Aspera Root element D suppress PTP1B activity.
One, experiment material and method.
Mentioned enzyme or reagent in this method, except specified otherwise, all purchased from Sigma-Aldrich company (U.S.).
Our bibliography set up PTP1B activity detection method (ElcheblyM, Science.1999,283,1544-1548). PTP1B can produce p-nitrophenol by catalysis 4-NPP (pNPP) dephosphorylation, and this product exists405nm has light absorption, can be used for monitoring PT P1B enzymatic activity. Concrete reaction buffer consists of the HEPES (pH=of 50mM7.2), the EDTA of 1mM and the dithiothreitol (DTT) of 5mM (DTT). Reaction is carried out in 96 orifice plates, each reaction system cumulative volumeBe 200 μ L, the PTP1B that comprises 0.1 μ g, the Radix Glycyrrhizae compound of the pNPP of 2mM and prescribed concentration. Reaction system is incubated at 37 DEG CEducate 30 minutes, then add the NaOH solution cessation reaction of 10 μ L10M, measure each hole extinction in 405nm with automatic ELIASADegree, and calculate its inhibiting rate with respect to control group.
Two, experimental result.
As shown in table 2, above-mentioned three kinds of compounds all approach 100% to the inhibiting rate of PTP1B under 25 μ M concentration, whereinThe IC of angustoneA to PTP1B50Value is 0.4 μ M. The above results illustrates that these compounds have significant pressing down to PTP1B activityMake use, can be used for preparing PTP1B inhibitor. Because PTP1B can pass through the junket on insulin receptor or its substrate in vivoPropylhomoserin residue dephosphorylation is transduceed and is carried out negative regulator insulin signaling, is therefore the important target spot of antidiabetic medicine,Above-mentioned reactive compound can be used for preparing antidiabetic medicine. In addition, PTP1B has also participated in cancer to be developed, and it presses downPreparation has great potential at anticancer aspect, and therefore the activity of above-claimed cpd inhibition PTP1B has also further supported their to useIn the purposes of preparing cancer therapy drug.
Embodiment 6, Glycyrrhiza Aspera Root element D restraint of tyrosinase activity.
One, experiment material and method.
Mentioned enzyme or reagent in this method, except specified otherwise, all purchased from Sigma-Aldrich company (U.S.).
Our bibliography set up tyrosinase activity test method (NeryaO etc.,Phytochemistry.2004,65,1389-1395). Concrete reaction buffer is the sodium phosphate buffer (pH=of 45mM6.6), reaction is carried out in 96 orifice plates. The cumulative volume of each reaction system is 200 μ L, the tyrosinase that comprises 9.99U, 1mM'sThe Radix Glycyrrhizae compound of TYR and prescribed concentration. Reaction system is hatched 15 minutes at 37 DEG C, then uses automatic ELIASAMeasure each hole absorbance in 492nm, calculate its its inhibiting rate with respect to control group; Further, thick according to variable concentrationsThe dose-effect relationship of hair glycyrrhizin D restraint of tyrosinase activity is calculated its IC50Value.
Two, experimental result.
As shown in table 2, Glycyrrhiza Aspera Root element D is 100% to the inhibiting rate of tyrosinase under 20 μ M concentration, its IC50Value onlyBe 0.1 μ M, illustrate that Glycyrrhiza Aspera Root element D has very strong inhibitory action to tyrosinase, can be used for preparing the inhibition of tyrosinaseAgent. Tyrosinase is the synthetic key enzyme of melanin in skin, and it is melanic synthetic in body that tyrosinase inhibitor can suppress,There is the effect of freckle removing and whitening. Therefore, Glycyrrhiza Aspera Root element D also can be used for preparing medicine or the skin care item of freckle removing and whitening.
Embodiment 7, licoricidin, angustoneA and Glycyrrhiza Aspera Root element D suppress AChE activity.
One, experiment material and method.
Mentioned enzyme or reagent in this method, except specified otherwise, all purchased from Sigma-Aldrich company (U.S.).
Our bibliography set up AChE activity test method (RheeIK etc., Phytochem.Anal.2003,14,145-149). Concrete reaction buffer is the Tris-HCl buffer solution (pH=8.0) of 30mM, and reaction is entered in 96 orifice platesOK. The cumulative volume of each reaction system is 200 μ L, comprises 6.25 × 10-5The AChE of U, 5 of 0.1mM, 5'-bis-thiobis (2-nitreYl benzoic acid) (DTNB), the acetylthiocholine iodide of 0.02mM (ATCI) and certain density Radix Glycyrrhizae compound. Reaction bodySystem at room temperature hatches 10 minutes, then measures each hole absorbance with automatic ELIASA in 405nm, calculates it with respect to control groupInhibiting rate; Further, calculate its IC according to the dose-effect relationship of the appointed compound inhibition AChE activity of variable concentrations50Value.
Two, experimental result.
As shown in table 2, above-mentioned three kinds of compounds all can significantly suppress AChE activity under 25 μ M concentration, and inhibiting rate all exceedes50%; The wherein inhibiting rate of licoricidin the highest (82%), its IC50Value is 15.5 μ M. The above results illustrates this three kinds of compoundsCan be used for preparing AChE inhibitor. AChE is optionally hydrolyzed in vivo acetylcholine and generates choline and acetic acid, reduces cholinergicNeural physiological effect, AChE inhibitor can be used as anticholinesterase medicine and is used for the treatment of nervous system degenerative disease as A ErThe diseases such as Zi Haimo disease; Therefore, above-mentioned three kinds of compounds can be used for preparing anticholinesterase medicine and are used for the treatment of relevant disease.
Table 2, licoricidin, angustoneA and Glycyrrhiza Aspera Root element D suppress the activity of PTP1B, tyrosinase and AChE.

Claims (5)

1. the compound shown in formula 1 to 3 or its pharmaceutically acceptable salt, ester, solvate, stereoisomer, tautomerismAt least one in body, prodrug or mixture are in the application of preparing in cancer therapy drug or antidiabetic medicine:
(1) licoricidin (licoricidin) (2) Glycyrrhiza Aspera Root element D(glyasperinD) (3) angustoneA.
2. application described in claim 1, it is characterized in that described anticancer be anti-liver cancer, lung cancer, intestinal cancer and/or breast cancer.
3. the compound shown in formula 1 to 3 or its pharmaceutically acceptable salt, ester, solvate, stereoisomer, tautomerismThe application in preparation PTP1B inhibitor of at least one in body, prodrug or mixture.
Compound or its pharmaceutically acceptable salt shown in formula 1 to 3, ester, solvate, stereoisomer, dynamic isomer,At least one in prodrug or mixture are in the application of preparing in acetylcholinesteraseinhibitors inhibitors or anticholinesterase medicine.
5. compound shown in formula 2 or its pharmaceutically acceptable salt, ester, solvate, stereoisomer, dynamic isomer, frontAt least one in medicine or mixture are in the application of preparing in tyrosinase inhibitor and freckle removing and whitening product.
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Cited By (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN109464438A (en) * 2019-01-14 2019-03-15 毕节市第人民医院 Glycyrrhiza Aspera Root element A is for the purposes in anti-tumor drug
CN110872267A (en) * 2018-08-30 2020-03-10 复旦大学 Compound extracted from paper mulberry and application of compound in preparation of protein tyrosine phosphatase 1B inhibitor
CN111329774A (en) * 2020-03-30 2020-06-26 香港科技大学 Novel use of acetylcholinesterase activity inhibitor
CN114028384A (en) * 2021-02-24 2022-02-11 中国医学科学院皮肤病医院(中国医学科学院皮肤病研究所) Application of licorice isoflavane derivative in preparation of medicine for preventing, relieving or/and treating pruritus
CN114848629A (en) * 2022-06-24 2022-08-05 中南大学湘雅二医院 Application of Liquiritidine in preparation of medicine or food for treating obesity and complications thereof

Citations (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
JP2003252784A (en) * 2002-02-27 2003-09-10 Kanegafuchi Chem Ind Co Ltd alpha-GLUCOSIDASE INHIBITOR
CN1599602A (en) * 2001-10-11 2005-03-23 钟渊化学工业株式会社 Peroxisome proliferator activated receptor ligands and process for producing the same
CN1633302A (en) * 2002-02-20 2005-06-29 钟渊化学工业株式会社 Method for producing high-quality hydrophobic licorice extract
KR20110050343A (en) * 2009-11-06 2011-05-13 한림대학교 산학협력단 Composition for anti-cancer or inhibiting a cancer metastasis comprising an extract of licorice as an active ingredient
CN104523940A (en) * 2014-12-19 2015-04-22 陕西东科制药有限责任公司 Preparation method for Ganhaiweikang preparation

Patent Citations (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN1599602A (en) * 2001-10-11 2005-03-23 钟渊化学工业株式会社 Peroxisome proliferator activated receptor ligands and process for producing the same
CN1633302A (en) * 2002-02-20 2005-06-29 钟渊化学工业株式会社 Method for producing high-quality hydrophobic licorice extract
JP2003252784A (en) * 2002-02-27 2003-09-10 Kanegafuchi Chem Ind Co Ltd alpha-GLUCOSIDASE INHIBITOR
KR20110050343A (en) * 2009-11-06 2011-05-13 한림대학교 산학협력단 Composition for anti-cancer or inhibiting a cancer metastasis comprising an extract of licorice as an active ingredient
CN104523940A (en) * 2014-12-19 2015-04-22 陕西东科制药有限责任公司 Preparation method for Ganhaiweikang preparation

Non-Patent Citations (3)

* Cited by examiner, † Cited by third party
Title
NAN KYOUNG LEE等: "Prenylated Flavonoids as Tyrosinase Inhibitors", 《ARCH PHARM RES》 *
SO YOUNG PARK等: "Hexane-Ethanol extract of Glycyrrhiza uralensis containing licoricidin inhibits the metastatic capacity of DU145 human prostate cancer cells", 《BRITISH JOURNAL OF NUTRITION》 *
ZHENZUO JIANG等: "Dose-dependent targeted knockout methodology combined withdeep structure elucidation strategies for Chinese licorice chemicalprofiling", 《JOURNAL OF PHARMACEUTICAL AND BIOMEDICAL ANALYSIS》 *

Cited By (8)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN110872267A (en) * 2018-08-30 2020-03-10 复旦大学 Compound extracted from paper mulberry and application of compound in preparation of protein tyrosine phosphatase 1B inhibitor
CN109464438A (en) * 2019-01-14 2019-03-15 毕节市第人民医院 Glycyrrhiza Aspera Root element A is for the purposes in anti-tumor drug
CN109464438B (en) * 2019-01-14 2020-12-29 毕节市第一人民医院 Application of glabridin A in antitumor drugs
CN111329774A (en) * 2020-03-30 2020-06-26 香港科技大学 Novel use of acetylcholinesterase activity inhibitor
WO2021197290A1 (en) * 2020-03-30 2021-10-07 香港科技大学 Novel application of acetylcholinesterase activity inhibitor
CN114028384A (en) * 2021-02-24 2022-02-11 中国医学科学院皮肤病医院(中国医学科学院皮肤病研究所) Application of licorice isoflavane derivative in preparation of medicine for preventing, relieving or/and treating pruritus
CN114028384B (en) * 2021-02-24 2023-05-23 中国医学科学院皮肤病医院(中国医学科学院皮肤病研究所) Application of liquorice isoflavan derivative in preparation of drug for preventing, relieving or/and treating pruritus
CN114848629A (en) * 2022-06-24 2022-08-05 中南大学湘雅二医院 Application of Liquiritidine in preparation of medicine or food for treating obesity and complications thereof

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