CN113072526A

CN113072526A - Anthraquinone/coumarin dimer novel skeleton compound and preparation method and application thereof

Info

Publication number: CN113072526A
Application number: CN202110339099.8A
Authority: CN
Inventors: 赵峰; 马丽; 王春华; 侯桂革; 曲兆霞
Original assignee: YANTAI STOMATOLOGICAL HOSPITAL; Binzhou Medical College
Current assignee: YANTAI STOMATOLOGICAL HOSPITAL; Binzhou Medical College
Priority date: 2021-03-30
Filing date: 2021-03-30
Publication date: 2021-07-06
Anticipated expiration: 2041-03-30
Also published as: CN113072526B

Abstract

The invention relates to an anthraquinone/coumarin dimer novel skeleton compound and a preparation method and application thereof. The structural formula of the dimer compound is as follows:

Description

Anthraquinone/coumarin dimer novel skeleton compound and preparation method and application thereof

Technical Field

The invention belongs to the field of pharmaceutical compounds, and particularly relates to a novel skeleton compound formed by coupling anthraquinone and coumarin through carbon-carbon bonds from euphorbia kankinensis, a preparation method of the compound, and application of the compound in preparation of PTP1B enzyme inhibitors, type 2 diabetes treatment medicines and/or antitumor medicines.

Background

Protein kinases and phosphatases are responsible for post-translational phosphorylation and dephosphorylation modifications of proteins, respectively, coordinate to regulate cell signaling pathways, maintain normal life activities, and cause various diseases such as cancer, immunity and metabolic system diseases when the balance is broken, so that the protein kinases and the phosphatases are important targets for drug development. Protein tyrosine phosphatase 1B (PTP1B) is a subtype of PTP family distributed in cells, is separated from human placenta tissue at first, and is widely distributed in tissues such as liver, kidney and placenta. PTP1B is closely related to insulin resistance and obesity, plays a role in negative regulation in insulin and leptin signaling pathways, and reduces the glucose and lipid metabolism capability of the body by regulating the levels of insulin and leptin through dephosphorylation of insulinotropic receptors. By inhibiting the activity of PTP1B, the sensitivity of peripheral tissues to insulin can be improved, so that PTP1B is an important target for treating type 2 diabetes and obesity. In recent years, the PTP1B is highly expressed in various tumor cells, and the inhibitor designed aiming at the PTP1B can obviously inhibit the proliferation of cells such as breast cancer, lung cancer and the like, and is a new target of antitumor drugs.

However, two characteristics of PTP1B greatly limit the development of inhibitors, one is that the catalytic active sites of PTPs are highly conserved, so that the selectivity of the inhibitors for PTP1B and other PTPs is low. For example, T-cell PTP (TCPTP) has approximately 80% of amino acid sequences of catalytic regions which are consistent, and the TCPTP plays an important role in hematopoiesis and immune systems, and has obvious side effects caused by cross inhibition of the TCPTP. The other is that the phosphate group of the substrate is electronegative, the active site of PTP1B is electropositive, and the inhibitor structure designed aiming at the active site has groups capable of dissociating negative charges, such as carboxyl, sulfonic acid group and the like, and the inhibitor synthesized by the simulated substrate has strong polarity, is difficult to penetrate through cell membranes, has poor bioavailability and seriously influences the drug success rate. 2004 reported an allosteric site of PTP1B approximately distant from the active site

Is composed of alpha 3 and alpha 6 helices, has low conservation, is mainly composed of lipophilic amino acids, and is bound to allosteric pocketShowing better physicochemical properties and remarkable selectivity. However, very few reports of PTP1B allosteric inhibitors have resulted in an extreme lack of lead compounds acting at the allosteric site. The chemical components in the natural products have unique molecular frameworks and spatial configurations, show various biological activities, and are important sources of innovative drugs and lead compounds with novel structures.

Knoxia valerioides is a perennial herb of the genus Knoxia in the family Rubiaceae, is distributed in various provinces in south China, mainly in Guangxi, and is distributed in small quantities in other areas such as Guangdong, Yunnan, Fujian, Guizhou and Hainan provinces. The plant root tuber has effects of purging water, removing fluid retention, removing toxic substance, relieving swelling, and resolving hard mass; it is used for treating hydrothorax and abdominal dropsy, constipation, carbuncle, sore, scrofula, and subcutaneous nodule. The researches of the inventor on the euphorbia for years show that the plant is rich in anthraquinone and triterpenoid components and small amounts of lignans, coumarins and phenolic acids, wherein the anthraquinone components have antiviral, anti-inflammatory and liver cell protection activities, and the research result provides material basis and scientific basis for the application of the euphorbia.

In the continuing research on Knoxia, the inventors discovered a novel framework compound, Knoxiquinone A, formed by the coupling of anthraquinones and coumarins via methylene bridges. The natural anthraquinone and coumarin parent nucleus structures are benzocyclodienone and benzo alpha-pyrone respectively, and the derivatives of the natural anthraquinone and coumarin parent nucleus structures mostly contain simple substituents such as hydroxyl, methoxyl and isopentenyl or are combined with sugar to form glycoside. The document reports a plurality of anthraquinone dimers and coumarin dimers, mainly including anthraquinone-anthraquinone, anthraquinone-kokurane, coumarin-coumarin, coumarin-chalcone dimers and other types, wherein the parent nuclei are coupled through phenyl and are connected through oxygen atoms in a connection mode, and the structure, the preparation method and the application report of the compound formed by coupling the anthraquinone-coumarin through methylene are not found. The compound has high-efficiency and high-selectivity PTP1B inhibitory activity, is a PTP1B allosteric inhibitor with a novel structure, can inhibit the proliferation of various liver cancer cells, and can be used for researching and developing novel hypoglycemic drugs with high specificity and good bioavailability, type 2 diabetes treatment drugs and/or antitumor drugs with a new mechanism and a new target.

Disclosure of Invention

The invention separates a new skeleton compound of anthraquinone and coumarin coupled by carbon-carbon bonds from the Knoxia by carrying out systematic chemical composition and biological activity research on the Knoxia.

One of the objects of the present invention is to provide a dimeric compound of anthraquinone-coupled coumarin, named as ingenol a, whose structure is shown below:

the compound is structurally characterized by containing anthraquinone and coumarin structural fragments, wherein the anthraquinone is in a benzo ring dienone structure, the coumarin is in a benzo alpha-pyrone structure or a benzo hexa-lactone ring structure, and the 2-methyl of the anthraquinone and the C-3 of the coumarin are coupled through carbon-carbon bonds to form a novel dimer compound.

Another object of the present invention is to provide a process for the preparation of the compound, comprising the steps of:

1) pulverizing radix Knoxiae, extracting with ethanol, and evaporating the extractive solution under reduced pressure to obtain extract;

2) dissolving the extract with water, extracting the water solution with ethyl acetate, evaporating ethyl acetate under reduced pressure to obtain ethyl acetate extraction part, and sequentially separating the ethyl acetate extraction part with silica gel column chromatography, MCI column chromatography and gel column chromatography to obtain part containing euphorbia pekinensis quinone A;

3) recrystallizing the part containing jolkinoquinone A with chloroform-methanol mixed solution, or purifying with high performance liquid chromatography to obtain jolkinoquinone A.

Preferably, in the step 1), the ethanol concentration is 95%, and the extraction temperature is normal temperature.

Preferably, in the step 2), silica gel column chromatography is performed by using 100-200 mesh silica gel and petroleum ether/acetone as mobile phase separation, and then using 200-300 mesh silica gel and chloroform/methanol as mobile phase separation. In MCI column chromatography, the stationary phase is MCI gel CHP20p, and the mobile phase is methanol/water. In the gel column chromatography, the gel is Sephadex LH-20, and the mobile phase is chloroform/methanol.

Preferably, in step 3), the ratio of chloroform/methanol in the recrystallization process is 3: 1.

Preferably, in step 3), in the high performance liquid chromatography, the chromatographic column is ODS packing, and the mobile phase is 80% methanol/water.

More preferably, in the step 2), the petroleum ether/acetone volume ratio used for silica gel column chromatography is 10:0, 10:1, 10:2, 2:1, 1:1 and 0:1, and the petroleum ether/acetone 2:1 elution part is collected, and the chloroform/methanol volume ratio used for silica gel column chromatography is 100:1, 10:1, 2:1, 1:1 and 0: 1; collecting chloroform/methanol 1:1 elution part, and separating by MCI column chromatography with methanol concentration of 30%, 50%, 70%, 90%, 100%; collecting 70% methanol/water eluate, and separating by gel column chromatography with chloroform/methanol volume ratio of 1: 1.

The invention also aims to provide application of the compounds in preparation of PTP1B inhibitors, type 2 diabetes treatment medicines and/or antitumor medicines.

The compound is a PTP1B allosteric inhibitor with a novel structure, and has high-efficiency and high-selectivity PTP1B inhibitory activity. IC for inhibition of PTP1B and TCPTP₅₀The values were 2.42. mu.M and 73.33. mu.M, respectively, with a selectivity greater than 30-fold. The results of enzyme inhibition kinetics experiments, molecular docking and experiments on the influence of glucose uptake capacity of C2C12 cells of skeletal muscle resisting insulin show that the compound is a mixed inhibitor of PTP1B, can be combined with PTP1B allosteric sites to form a stable molecule-protein complex, and can improve the glucose consumption capacity of the skeletal muscle cells resisting insulin. Under the condition that few reports of PTP1B allosteric inhibitors are reported, the compound disclosed by the invention provides a novel structural template, provides reference for further synthesis and structural modification to develop a PTP1B inhibitor with high specificity and good bioavailability, and has a good application prospect in the aspects of developing novel hypoglycemic drugs targeting PTP1B, type 2 diabetes therapeutic drugs and/or antitumor drugs.

Drawings

FIG. 1: results of enzyme inhibition kinetics experiments

FIG. 2: molecular docking results

FIG. 3: changes in glucose uptake in insulin resistant skeletal muscle C2C12 cells

Detailed Description

The compounds, preparation steps and activity screening experimental procedures of the present invention are further illustrated by the following specific examples, which are used to illustrate the technical content of the present invention and are not intended to limit the content of the present invention, and all changes or equivalents based on the present invention shall fall within the protection scope of the present invention.

Example 1 preparation of Knoxiquinone A

20Kg of euphorbia pekinensis, crushing, performing ultrasonic extraction with 95% ethanol at normal temperature, combining extracting solutions, and evaporating the solvent under reduced pressure to obtain 3.9Kg of extract. Dissolving the extract in water, extracting with ethyl acetate for 3 times, and evaporating to remove solvent to obtain 400g of ethyl acetate extract. Separating ethyl acetate part with silica gel column chromatography, sequentially eluting with petroleum ether/acetone at volume ratio of 10:0, 10:1, 10:2, 2:1, 1:1, and 0: 1. The petroleum ether/acetone 2:1 eluate fraction (30g) was again subjected to silica gel column chromatography, and eluted sequentially with chloroform/methanol at volume ratios of 100:1, 10:1, 2:1, 1:1, and 0: 1. The chloroform/methanol 1:1 eluate (4.7g) was separated by MCI column chromatography, followed by elution with 30%, 50%, 70%, 90%, 100% methanol/water. Evaporating 70% methanol/water eluate under reduced pressure, dissolving with small amount of chloroform/methanol, purifying with Sephadex LH-20 gel column chromatography, eluting with chloroform/methanol (V/V)1:1 as eluting solvent, analyzing the eluate with thin layer chromatography, mixing, evaporating to obtain orange solid, and recrystallizing one part with appropriate amount of chloroform/methanol (V/V)1:1 to obtain Euphorbia Kishinouquinone A (3.1 mg). The remaining fractions were purified by HPLC at a flow rate of 2.5mL/min, detection wavelength of 280nm, mobile phase 80% methanol/water to give ingenol A (15.3 mg). The structure was identified by the following spectral information and physicochemical properties.

Euphorbia quinone a (valeraquinone a): an orange amorphous powder; UV (MeOH) max (log. epsilon.) 204(6.2),239(4.4),281(4.3),336(3.6),407(3.3) nm; IR max 3743(OH),3304(OH),3216(OH),2924,2854,1663(C ═ O),1590,1512,1462,1396,1365,1331,1301,1280,1163,1120,1092,1030,994,953,829,790,712cm^-1；¹H NMR(DMSO-d₆,500MHz)δ7.37(1H,s,H-4),8.19(1H,dd,J＝7.0,1.75,H-5),7.95(1H,dd,J＝7.0,7.0,1.95,H-6),7.93(1H,dd,J＝7.0,7.0,1.75,H-7),8.24(1H,dd,J＝7.0,1.95,H-8),3.77(2H,d,J＝3.77,H-11),7.18(2H,t,J＝3.77,H-4′),6.68(1H,s,H-5′),13.18(1H,s,1-OH),11.47(1H,s,3-OH),3.70(3H,s,6′-CH₃),9.36(1H,brs,7′-OH),9.41(1H,brs,8′-OH)；¹³C NMR(DMSO-d₆,125MHz)δ163.3(C-1),117.7(C-2),163.9(C-3),108.2(C-4),133.5(C-4a),127.3(C-5),135.2(C-6),135.1(C-7),126.9(C-8),133.3(C-8a),186.8(C-9),109.8(C-9a),182.4(C-10),133.5(C-10a),23.4(C-11),161.8(C-2′),121.9(C-3′),139.0(C-4′),100.3(C-5′),145.8(C-6′),138.7(C-7′),133.1(C-8′),138.4(C-9′),111.0(C-10′),156.3(4′-OCH3)；(-)-HRESIMS m/z 459.0710(calcd for C25H15O9,459.0722).

Example 2 assessment of PTP1B and TCPTP inhibitory Activity

Compounds were formulated in DMSO at gradient concentrations (

final concentrations

50, 25, 12.5, 6.2, 3.1, 1.6 μ M, respectively). A pre-prepared enzyme solution (buffer containing 25mM Hepes pH 7.5,150mM NaCl,5mM DTT,2mM EDTA, containing 0.1. mu.g PTP1B or 0.15. mu.g TCPTP) was added to a 96-well plate at 90. mu.L per well, 1. mu.L each of the compound solution was added, with DMSO as a blank, three parallel duplicate wells per concentration, and the 96-well plate was placed in a constant temperature shaker and shaken at 37 ℃ for 15 min. After 15min, 10. mu.L of a final 4mM p-NPP solution were added and shaken for 30min at 37 ℃ in a constant temperature shaker. Taking out after 30min, sequentially adding 20 μ L of 3M NaOH solution to terminate reaction, measuring OD value at 405nm of microplate reader, and calculating the inhibition rate of the compound on PTP1B and IC₅₀The value is obtained. Ursolic acid is used as positive control drug.

Calculating the formula: inhibition rate (OD)_{Blank space}-OD_{Sample (I)})/OD_{Blank space}×100％

Statistical analysis is carried out on the data by using GraphPad Prism 8.0 software, three independent experiments are repeated, and the data are represented by means of the average number +/-standard deviation.

TABLE 1 inhibitory Effect of Knoxiquinone A on PTP1B and TCPTP

The results in table 1 show that ingenol a has a significant inhibitory effect on PTP1B, is stronger than ursolic acid, which is a control drug, and has a very weak effect on TCPTP, showing selective inhibitory activity on PTP 1B.

EXAMPLE 3 kinetics of PTP1B enzyme inhibition experiment

Adding a prepared PTP1B solution into a 96-well plate, adding 90 mu L of each well, adding 1 mu L of a compound solution to be detected respectively, setting three multiple wells for each concentration, taking DMSO as a blank control, putting the 96-well plate into a constant temperature oscillator, and shaking for 15min at 37 ℃. Then, p-NPP solution (final concentration is 2, 4 and 8mM respectively) is added, the change of OD value within 15min at 405nm is immediately detected by a microplate reader, and the initial rate of enzymolysis is calculated. The inhibition constant Ki value was obtained by plotting the compound concentration and the initial rate of enzymatic hydrolysis by Dixon plot, the slope of the line at each concentration obtained by Lineweaver-Burk plot, and the compound concentration and slope. Compounds were analyzed for the type of inhibition of PTP 1B.

FIG. 1 shows the results of kinetic experiments, in which the sets of curves in FIG. 1(A) intersect in the xy-axis region, indicating that Knoxiquinone A is a PTP1B mixed inhibitor, capable of binding to sites other than the catalytically active region of PTP 1B.

Example 4 molecular docking

Molecular docking simulation of the active site and allosteric site of the crystal structure of ingequinone A and PTP1B was performed using the C-DOCKER module of the molecular docking software BIOVIA Discovery Studio 3.0. The crystal structure of PTP1B is from a protein crystal library, numbered 2HB1 (crystal formed by binding a substrate to the PTP1B active site), 1T49 (crystal formed by binding a compound to the PTP1B allosteric site). The 3D structure of the ingenol a was first calculated using Discovery Studio 3.0 software to obtain the optimal conformation. PTP1B protein structure was optimized with CHARM force field structure after dehydration and hydrogenation. Then, a binding site is defined based on a receptor site, after the receptor is removed, the optimal conformation of the compound is subjected to butt joint calculation with the crystal structure of PTP1B, scoring is carried out through software, and the action site and the binding mode of the highest-scoring conformation are analyzed.

FIG. 2 shows the results of molecular docking, whereby ingenol A successfully docks with the active site of 2HB1, the allosteric site of 1T 49. FIG. 2(a, b, c) is the overall molecule-protein binding map after docking with the active site (2a), the docking map with the active site (2b) and the two-dimensional map of the mode of action (2c), respectively. Knoxiquinone A only forms weak pi-alkyl interactions with Ala217 in the catalytically active site P-loop (residues 215-222), but has no effect with key amino acid residues Cys215 and Arg 221. Can act with other sites nearby the active site, including pi-pi accumulation and hydrogen bond with Tyr46 in pTyr-loop (residues 47-49) for recognition and combination of auxiliary substrates, hydrogen bond with Lys120 in Lys-loop (residues 119-121), and NH of Gln262 in Q-loop (residues 261-262) of the second combination site₂And (4) interaction. It has been reported that inhibitors that bind to pTyr-loop, Lys-loop and Q-loop show better selectivity for PTP 1B. FIGS. 2(d, e, f) are a graph of overall molecule-protein binding after docking with an allosteric site (2d), a graph of docking with an active site (2e), and a two-dimensional graph of the mode of action (2f), respectively, and show that the ingenol A binds well to an allosteric site, that the anthraquinone fragment is inserted into the hydrophobic pocket formed by Phe280 and Phe196 in the alpha 3 and alpha 6 helices, that three loops in the structure form strong pi-pi stacking interactions with both, and that the lactone loop of coumarin also forms pi-stacking interactions with the non-conserved, key amino acid residue Phe280 in the allosteric site (Cys 280 in TCPTP, which is the position of cysteine 276), and that overall a compound assumes a conformation surrounding Phe280, forming a stable molecule-protein complex and further stabilizing the complex in the inactive conformation through hydrogen bonding with 193 Asn, hydrogen bonding with Gln276, and hydrophobic interactions with Ala189, disabling PTP1B from undergoing a catalytically active conformational transition. The overall molecular docking result shows that the ingenol A has weak ability to compete with the substrate for the active site, and has better binding effect with the allosteric site, which is probably the main reason of the selective inhibition effect of the ingenol A, and the ingenol A is an allosteric inhibitor with novel structure and can be developed into a high-efficiency and high-selectivity PTP1B inhibitor.

Example 5 evaluation of tumor cytotoxic Activity

The cytotoxic activity of the ingenol A on three human liver cancer HepG2, SMMC-7721 and QGY-7703 cells was tested by MTT method. After the tumor cells are recovered, the cells are subcultured by a DMEM medium containing 10% fetal calf serum and 1% double antibody, and the cells in the logarithmic growth phase are taken for experiments. The final concentrations of the compounds were 50, 25, 12.5, 6.25, 3.13 μ M in order. Taking the well-grown cells, according to 2X 10⁴Per 100. mu.l of plate; after 12h incubation, the supernatant was discarded and each concentration of drug was added to the cells in a volume of 100. mu.l per well, with three replicate wells per concentration. Adding drugs for culturing for 24h, removing the supernatant, adding 100 mul of MTT solution prepared by serum-free culture medium into each hole, continuously culturing for 4h, removing the supernatant, adding 150 mul of DMSO into each hole, shaking on a microplate oscillator for 10min, measuring the OD value at 570nm by using an enzyme-labeling instrument, and calculating the cell survival rate. Cell viability was defined as OD value of experimental group/OD value of control group × 100%. The experiment was repeated three times.

TABLE 2 cytotoxic Activity of Knoxiquinone A

The test result shows that the euphorbia pekinensis quinone A has obvious anti-tumor activity, wherein the inhibition effect on HepG2 is higher than that of the positive control drug adriamycin.

EXAMPLE 6 Effect of Compounds on insulin resistant skeletal muscle C2C12 cell glucose consumption

Cell differentiation: C2C12 mouse myoblasts were passaged when cultured to 80% density. After culturing a sufficient number of cells, the cells were digested, centrifuged, resuspended at 2X 10⁴Plating each cell per 100 mul, culturing in a cell culture box for 12h, removing supernatant after the cells adhere to the wall, adding 100 mul PBS to each hole for rinsing three times, adding 200 mul fresh differentiation culture solution to each hole, and continuously culturing for 24 h. From then on, the fresh differentiation medium was replaced every 24h and co-differentiation culture was carried out for 96h until single spindle cells were differentiated into regularly arranged multinuclear fused myocytes.

Construction of muscle cell model for insulin resistance: modeling method for referenceThe method comprises removing differentiated muscle cell culture solution, adding 100 μ l PBS to each well, rinsing cells for 3 times, adding 200 μ l high-glucose DMEM culture medium to each well of normal group cells, adding 200 μ l prepared modeling insulin solution to each well of model group and drug-added group cells, placing cells at 37 deg.C and 5% CO₂The cell culture box is continuously cultured for 48 hours. After 48h, if the model group is compared with the blank group, the glucose uptake of the cells is obviously reduced, and the success of modeling is proved.

Administration: diluting the compound with high-glucose DMEM culture medium to final concentrations of 50 muM, 33.3 muM and 22.2 muM in sequence, sucking and discarding culture solution in each well, adding 200 muL of high-glucose DMEM culture medium into each well of cells of a normal group and a model group, adding 200 muL of drugs with various concentrations into each well of a dosing group, setting three multiple wells for each concentration, and repeating three independent experiments.

And (3) detecting the content of glucose: the cells were placed in an incubator for further culture. After 48h, preparing a glucose content detection solution according to the operation method of the instruction in the kit, adding 180 mul of the glucose content detection solution into each hole of a new 96-hole plate, respectively adding 20 mul of cell supernatant of each hole, and performing shake incubation for 15min in a constant-temperature shaker at 37 ℃. After 15min, the OD value at 505nm is measured on a microplate reader by using a 96-well plate, and the glucose content in each well is calculated according to a calculation formula of a kit specification. 20 mul of MTT solution prepared in advance is respectively added into the supernatant of a 96-well plate containing cells, and after the supernatant is put into a cell incubator to be cultured for 4 hours, the OD value at 490nm is measured by an enzyme-labeling instrument. To remove the influence of the cell number on the assay results, the glucose content/MTT value was used as the final glucose content.

FIG. 3 is a graph of the change in glucose consumption by insulin resistant skeletal muscle C2C12 cells after treatment with different concentrations of ingenol A. Compared with the normal group, the cells of the model group have obviously reduced glucose consumption, while the cells of the administration group have obviously increased glucose consumption, and the glucose consumption at medium and high concentrations is even close to that of the normal group. Therefore, the compound can remarkably reverse the glucose consumption capability of insulin-resistant cells, and is expected to be developed into a novel hypoglycemic medicament or a medicament for treating type 2 diabetes.

Claims

1. An anthraquinone/coumarin dimer novel framework compound has the following structure:

the molecular formula is C₂₅H₁₆O₉Named as: knoevenagene a, english name: valeriaquinone A.

2. The novel anthraquinone/coumarin dimer scaffold compound as claimed in claim 1, wherein the scaffold structure is formed by coupling anthraquinone and coumarin through carbon-carbon bonds, wherein anthraquinone is a benzo ring dienone structure, coumarin is a benzo α -pyrone structure or a benzo six-membered lactone ring structure, and the linking group between the two is methylene. The anthraquinone fragment is substituted by hydroxyl, and the coumarin fragment is substituted by hydroxyl and methoxyl.

3. The preparation method of the anthraquinone/coumarin dimer new skeleton compound as claimed in claim 1, which comprises the following steps:

(1) extraction: pulverizing Euphorbiae radix, extracting with ethanol, filtering to obtain extractive solution, and concentrating to obtain extract;

(2) separation: dissolving the extract with water, extracting the water solution with ethyl acetate, evaporating ethyl acetate under reduced pressure to obtain ethyl acetate extraction part, and sequentially separating the ethyl acetate extraction part with silica gel column chromatography, MCI column chromatography and gel column chromatography to obtain part containing euphorbia pekinensis quinone A;

(3) and (3) purification: recrystallizing the part containing jolkinoquinone A with chloroform-methanol mixed solution, or purifying with high performance liquid chromatography to obtain jolkinoquinone A.

4. The method for preparing a novel anthraquinone/coumarin dimer scaffold compound according to claim 3, wherein in the step (1), the concentration of ethanol is 95%, and the extraction temperature is normal temperature.

5. The method for preparing a novel anthraquinone/coumarin dimer skeleton compound as claimed in claim 3, wherein in the step (2), silica gel column chromatography is performed by using 100-200 mesh silica gel and petroleum ether/acetone as mobile phase separation, and then using 200-300 mesh silica gel and chloroform/methanol as mobile phase separation. In MCI column chromatography, the stationary phase is MCI gel CHP20p, and the mobile phase is methanol/water. In the gel column chromatography, the gel is Sephadex LH-20, and the mobile phase is chloroform/methanol.

6. The process for preparing a novel skeleton compound of anthraquinone/coumarin dimer according to claim 3, wherein in the step (3), the ratio of chloroform/methanol in the recrystallization method is 3: 1.

7. The process for preparing a novel skeleton compound of anthraquinone/coumarin dimer as claimed in claim 3, wherein in step (3), the high performance liquid chromatography column is ODS filler, and the mobile phase is 80% methanol/water.

8. Use of an anthraquinone/coumarin dimer compound according to claim 1 in the preparation of a PTP1B enzyme inhibitor.

9. The use of an anthraquinone/coumarin dimer compound according to claim 1 in the preparation of a medicament for the treatment of type 2 diabetes.

10. The use of an anthraquinone/coumarin dimer compound as claimed in claim 1 in the preparation of an anti-tumor medicament.