Chalcone derivative, preparation method thereof and application of chalcone derivative in preparation of anti-angiogenesis medicine

Abstract

The invention relates to a chalcone derivative, a preparation method thereof and application of the chalcone derivative in preparation of an anti-angiogenesis medicine. The molecular structure of the chalcone derivative is shown as a formula I. The chalcone derivative disclosed by the invention can obviously inhibit angiogenesis in a vascular endothelial cell HMEC-1 and a zebra fish body at a relatively low concentration, and can be used as a new chemical entity for preparing a novel anti-angiogenesis medicine.

Description

Technical Field

[0001] This invention belongs to the field of pharmaceutical technology, and relates to natural product drugs, specifically to a chalcone derivative, its preparation method, and its application in the preparation of anti-angiogenic drugs. Background Technology

[0002] Angiogenesis plays a crucial role in the development and progression of various diseases, such as malignant tumors, rheumatoid arthritis, diabetic retinopathy, psoriasis, and atherosclerotic diseases. Especially in oncology, abnormally activated angiogenesis not only provides tumors with essential oxygen and nutrients but also promotes their proliferation, invasion, and distant metastasis, becoming a key factor in the formation and maintenance of the tumor microenvironment. Since Judah Folkman proposed anti-angiogenesis as a tumor treatment strategy in the 1970s, this field has rapidly developed into one of the core directions of cancer treatment. Currently, widely used anti-angiogenic drugs in clinical practice mainly fall into two categories: one is monoclonal antibodies targeting vascular endothelial growth factor (VEGF) or its receptors, such as bevacizumab; the other is multi-target small molecule tyrosine kinase inhibitors, such as sunitinib. However, existing drugs still face many challenges, including primary or secondary drug resistance, significant toxic side effects such as hypertension and proteinuria, and problems such as pathway compensatory activation due to single-target targeting, which severely limit their long-term efficacy and clinical application. Therefore, the development of novel anti-angiogenic drugs has become an important research direction and an urgent clinical need. Summary of the Invention

[0003] Therefore, the purpose of this invention is to provide a novel anti-angiogenic drug.

[0004] The technical solutions for achieving the above objectives include the following.

[0005] In a first aspect, the present invention provides a chalcone derivative or a pharmaceutically acceptable salt thereof or a stereoisomer thereof, the molecular structure of which is shown in Formula I.

[0006] .

[0007] Secondly, the present invention provides the use of the chalcone derivatives, or pharmaceutically acceptable salts thereof, or stereoisomers thereof, in the preparation of medicaments for inhibiting angiogenesis.

[0008] Thirdly, the present invention provides a drug for inhibiting angiogenesis, which is prepared from an active ingredient and pharmaceutically acceptable excipients, wherein the active ingredient comprises the chalcone derivatives described in the present invention or pharmaceutically acceptable salts thereof or stereoisomers thereof.

[0009] Fourthly, the present invention provides a method for preparing the chalcone derivative, comprising the following steps:

[0010] (1) Crush the branches and leaves of the guarful tree, extract them by percolation with an ethanol aqueous solution, concentrate the resulting extract to obtain a crude extract;

[0011] (2) The crude extract is suspended in water and then extracted with petroleum ether. The resulting extract is concentrated to obtain the petroleum ether extract fraction.

[0012] (3) The petroleum ether extract is separated and purified to obtain the chalcone derivative.

[0013] The present invention has the following beneficial effects:

[0014] (1) The present invention extracts, purifies and separates a novel chalcone derivative from Guafu wood. The compound is an oligomer formed by the combination of 2 chalcone fragments and 2 phenylpropionyl fragments, and has a 6 / 5-5 / 6 ring system. It is a chemical entity with a novel skeleton structure.

[0015] (2) The chalcone derivative prepared in this invention can significantly inhibit the formation of new blood vessels in vascular endothelial cells HMEC-1 and zebrafish at low concentrations, and can be used as a new chemical entity to prepare novel anti-angiogenic drugs. Attached Figure Description

[0016] Figure 1 Chalcone derivatives 1 H NMR spectrum.

[0017] Figure 2 Chalcone derivatives 13 C NMR spectrum.

[0018] Figure 3 Chalcone derivatives 1 H- 1 H COSY diagram.

[0019] Figure 4 This is the HSQC diagram for chalcone derivatives.

[0020] Figure 5 The image shows the HMBC diagram of chalcone derivatives.

[0021] Figure 6 The NOESY diagram is for chalcone derivatives.

[0022] Figure 7 This is a single-crystal X-ray diffraction structure diagram of a chalcone derivative.

[0023] Figure 8The results show the anti-angiogenic activity of chalcone derivatives against HMEC-1 vascular endothelial cells.

[0024] Figure 9 The results show the anti-angiogenic activity of chalcone derivatives in zebrafish. Detailed Implementation

[0025] To facilitate understanding of the present invention, a more complete description will be provided below. The present invention can be implemented in many different forms and is not limited to the embodiments described herein. Rather, these embodiments are provided to provide a thorough and complete understanding of the disclosure of the present invention.

[0026] Unless otherwise specified, experimental methods in the following examples are generally performed under standard conditions or as recommended by the manufacturer. All commonly used chemical reagents used in the examples are commercially available products.

[0027] Unless otherwise defined, all technical and scientific terms used in this invention have the same meaning as commonly understood by one of ordinary skill in the art to which this invention pertains. The terminology used in this specification is for the purpose of describing particular embodiments only and is not intended to limit the invention. The term "and / or" as used in this invention includes any and all combinations of one or more of the associated listed items.

[0028] Furthermore, as used herein, the term "or" is an inclusive "or" sign and is equivalent to the term "and / or" unless the context clearly specifies otherwise. The term "based on" is not exclusive and allows for basing on other factors not described unless the context clearly specifies otherwise. Additionally, throughout the specification, the meanings of "an," "a," and "the" include plural indicators. The meaning of "in" includes both "in" and "on."

[0029] Some embodiments of the present invention relate to a chalcone derivative or a pharmaceutically acceptable salt thereof or a stereoisomer thereof, the molecular structure of which is shown in Formula I.

[0030] .

[0031] Some embodiments of the present invention relate to the use of the chalcone derivatives, or pharmaceutically acceptable salts thereof, or stereoisomers thereof, in the preparation of medicaments for inhibiting angiogenesis.

[0032] The angiogenesis inhibitors described in this invention can be drugs that inhibit tumor angiogenesis, drugs that inhibit angiogenesis in rheumatoid arthritis lesions, drugs that inhibit angiogenesis in diabetic retinopathy lesions, drugs that inhibit angiogenesis in psoriasis lesions, or drugs that inhibit angiogenesis in atherosclerotic lesions, etc.

[0033] Some embodiments of the present invention relate to a medicament for inhibiting angiogenesis, which is prepared from an active ingredient and pharmaceutically acceptable excipients, said active ingredient comprising the chalcone derivatives described in the present invention or pharmaceutically acceptable salts thereof or stereoisomers thereof.

[0034] The amount of active ingredients contained therein is within the safe and effective range. "Safe and effective range" means that the amount of active ingredients is sufficient to significantly improve the condition without causing serious side effects.

[0035] "Pharmaceutical acceptable excipients" refer to one or more compatible solid or liquid fillers or gelling substances that are suitable for human use and must have sufficient purity and sufficiently low toxicity.

[0036] "Compatibility" here refers to the ability of the components in the composition or formulation to interact with and incorporate with the active ingredient of the present invention without significantly reducing the efficacy of the active ingredient.

[0037] Pharmaceutically acceptable examples of excipients (or carriers) include cellulose and its derivatives (such as sodium carboxymethyl cellulose, sodium ethyl cellulose, cellulose acetate, etc.), gelatin, talc, solid lubricants (such as stearic acid, magnesium stearate), calcium sulfate, vegetable oils (such as soybean oil, sesame oil, peanut oil, olive oil, etc.), polyols (such as propylene glycol, glycerin, mannitol, sorbitol, etc.), emulsifiers (such as Tween®), wetting agents (such as sodium dodecyl sulfate), colorants, flavoring agents, stabilizers, antioxidants, preservatives, pyrogen-free water, etc.

[0038] The administration method of the active ingredient, pharmaceutical composition, or pharmaceutical preparation of the present invention is not particularly limited. Representative administration methods include, but are not limited to, oral, transdermal, rectal, and parenteral (intravenous, intramuscular, or subcutaneous) administration. That is, the dosage form of the pharmaceutical preparation includes, but is not limited to, capsules, granules, tablets, pills, powders, drops, ointments, patches, liniments, sprays, powders, suppositories, sustained-release preparations, and injections.

[0039] Solid dosage forms for oral administration include capsules, granules, tablets, pills, and powders. In these solid dosage forms, the active ingredient is mixed with at least one conventional inert excipient (or excipient or carrier), such as sodium citrate or dicalcium phosphate, or with the following components:

[0040] (a) Fillers or compatibilizers, such as starch, lactose, sucrose, glucose, mannitol and silica;

[0041] (b) Adhesives, such as hydroxymethylcellulose, alginate, gelatin, polyvinylpyrrolidone, sucrose and gum arabic;

[0042] (c) Moisturizers, such as glycerin;

[0043] (d) Disintegrants, such as agar, calcium carbonate, potato starch or tapioca starch, alginate, certain complex silicates, and sodium carbonate;

[0044] (e) Slow solvents, such as paraffin;

[0045] (f) Absorption accelerators, for example, quaternary ammonium compounds;

[0046] (g) Wetting agents, such as cetyl alcohol and glyceryl monostearate;

[0047] (h) Adsorbents, such as kaolin; and

[0048] (i) Lubricants, such as talc, calcium stearate, magnesium stearate, solid polyethylene glycol, sodium dodecyl sulfate, or mixtures thereof. In capsules, tablets, and pills, the dosage form may also contain a buffer.

[0049] The solid dosage form can also be prepared using coatings and shells, such as casings and other materials known in the art. They may contain opacifying agents, and the release of the active ingredient from this composition can be delayed in a portion of the digestive tract. Examples of suitable encapsulating components are polymers and waxes.

[0050] Liquid dosage forms for oral administration include pharmaceutically acceptable emulsions, solutions, suspensions, syrups, or tinctures. In addition to the active ingredient, liquid dosage forms may contain inert diluents conventionally used in the art, such as water or other solvents, solubilizers and emulsifiers, e.g., ethanol, isopropanol, ethyl carbonate, ethyl acetate, propylene glycol, 1,3-butanediol, dimethylformamide, and oils, particularly cottonseed oil, peanut oil, corn germ oil, olive oil, castor oil, and sesame oil, or mixtures thereof. Besides these inert diluents, the composition may also contain adjuvants such as wetting agents, emulsifiers and suspending agents, sweeteners, flavoring agents, and fragrances.

[0051] In addition to the active ingredient, the suspension may contain suspending agents, such as ethoxylated isooctadecyl alcohol, polyoxyethylene sorbitol and dehydrated sorbitol esters, microcrystalline cellulose, aluminum methoxide and agar, or mixtures of these substances.

[0052] Compositions for parenteral injection may comprise physiologically acceptable sterile aqueous or anhydrous solutions, dispersions, suspensions, or emulsions, and sterile powders for reconstitution into sterile injectable solutions or dispersions. Suitable aqueous and non-aqueous carriers, diluents, solvents, or excipients include water, ethanol, polyols, and suitable mixtures thereof.

[0053] The chalcone derivatives described in this invention can be extracted from the branches and leaves of the guarberry tree, preferably by the following preparation method.

[0054] Some embodiments of the present invention also relate to a method for preparing the chalcone derivatives described herein, comprising the following steps:

[0055] (1) Crush the branches and leaves of the guarful tree, extract them by percolation with an ethanol aqueous solution, concentrate the resulting extract to obtain a crude extract;

[0056] (2) The crude extract is suspended in water and then extracted with petroleum ether. The resulting extract is concentrated to obtain the petroleum ether extract fraction.

[0057] (3) The petroleum ether extract is separated and purified to obtain the chalcone derivative.

[0058] In some embodiments of the present invention, the volume concentration of the ethanol aqueous solution is 90%-95%.

[0059] In some embodiments of the present invention, the percolation extraction in step (1) is performed 3-5 times.

[0060] In some embodiments of the present invention, the time for each percolation extraction is 40-56 hours.

[0061] In some embodiments of the present invention, the ratio of ethanol aqueous solution to Guafu wood used in each percolation extraction is 1.6L-2.4L:1kg.

[0062] In some embodiments of the present invention, the ratio of crude extract to water in step (2) is 1 kg: 3.5 L - 4.5 L.

[0063] In some embodiments of the present invention, in step (2), the extraction is performed 4-6 times, and the volume ratio of petroleum ether to water used each time is 0.8-1.2:1.

[0064] In some embodiments of the present invention, the separation and purification in step (3) includes: sequentially separating and purifying the petroleum ether extract by silica gel column chromatography, reversed ODS column chromatography, Sephadex LH-20 column chromatography, and reversed preparative HPLC.

[0065] In some embodiments of the present invention, the eluent for the silica gel column chromatography is petroleum ether and ethyl acetate, preferably eluted according to an elution gradient of petroleum ether to ethyl acetate volume ratios of 100:0, 1000:1, 1000:3, 100:5, 100:1, 100:3, 100:5, 100:10, 100:20, 100:50, 100:100, and 0:100.

[0066] In some embodiments of the present invention, the eluent for the reversed ODS column chromatography is methanol and water, preferably eluted according to an elution gradient of methanol to water volume ratios of 40:60, 50:50, 60:40, 70:30, 75:25, 80:20, 85:15, 90:10, 95:5, and 100:0.

[0067] In some embodiments of the present invention, the eluent used for separation and purification by the Sephadex LH-20 column is a mixed solvent of dichloromethane and methanol in a volume ratio of 1:0.9-1.1, and the preferred flow rate is 0.8 mL / min-1.2 mL / min.

[0068] In some embodiments of the present invention, the conditions for the reverse preparative HPLC separation and purification include: a Cosmosil C18 column, an eluent of acetonitrile and water in a volume ratio of 85-90:15-10, and a flow rate of 2.8 mL / min-3.2 mL / min.

[0069] The present invention will be further described in detail below with reference to specific embodiments.

[0070] Example 1: Preparation of Chalcone Derivatives

[0071] S1. Crush 23 kg of Guafu wood branches and leaves, and extract them four times with 95% (v / v) ethanol aqueous solution. The ratio of ethanol aqueous solution to Guafu wood is 2.0 L: 1 kg each time, and each time for 48 hours. Combine the extracts and concentrate them by vacuum distillation until there is no alcohol taste to obtain 2.6 kg of crude extract.

[0072] S2. The crude extract obtained in step S1 was added to distilled water at a ratio of 4 L of water to 1 kg of crude extract to obtain an aqueous suspension. Then, petroleum ether was added at a volume ratio of 1:1 to water for extraction, and the extraction was performed 5 times. The extracts were combined and concentrated under reduced pressure to obtain 554 g of the petroleum ether extract fraction.

[0073] S3. The petroleum ether extract obtained in step S2 was subjected to silica gel column chromatography with petroleum ether-ethyl acetate as the eluent. Elution was carried out according to the volume ratio of petroleum ether to ethyl acetate of 100:0, 1000:1, 1000:3, 100:5, 100:1, 100:3, 100:5, 100:10, 100:20, 100:50, 100:100 and 0:100. After analysis by thin-layer chromatography (TLC), similar fractions were combined to obtain 14 main fractions Fr. 1 to Fr. 14.

[0074] S4. The fraction Fr. 11 (26.7 g) obtained in step S3 was concentrated under reduced pressure and then subjected to reverse ODS column chromatography. The methanol-water system was used as the eluent, and elution was carried out according to the elution gradient of methanol to water volume ratio of 40:60, 50:50, 60:40, 70:30, 75:25, 80:20, 85:15, 90:10, 95:5 and 100:0 to obtain 10 fractions Fr. 11A~Fr. 11J.

[0075] S5. Collect the eluent fraction Fr. 11J obtained in step S4 with a methanol:water volume ratio of 100:0, concentrate it under reduced pressure, and load it onto a Sephadex LH-20 column. Elute with a dichloromethane-methanol solution with a volume ratio of 1:1 at a flow rate of 1 mL / min. Collect the eluent and analyze it by TLC to obtain 7 fractions Fr. 11Ja~Fr. 11Jg.

[0076] S6. The fifth fraction Fr. 11Je obtained in step S5 was concentrated under reduced pressure, dissolved in methanol, and then purified by reverse preparative HPLC. The chromatographic column was a Cosmosil C18 (10 × 250 mm, 5 μm) column, with acetonitrile-water at a volume ratio of 85:15 as the eluent and a flow rate of 3 mL / min. The eluent with a retention time of 28.5 min was collected and concentrated to obtain the chalcone derivative (colorless flaky crystals, 9.80 mg).

[0077] Example 2: Structural Identification of Chalcone Derivatives

[0078] The chalcone derivatives extracted and separated in Example 1 were detected by ultraviolet, infrared, high-resolution mass spectrometry, nuclear magnetic resonance and single-crystal X-ray diffraction, and the following experimental data were obtained:

[0079] UV (CH3CN) λmax (log ε) 237 (3.33), 285 (3.28) nm;

[0080] IR (KBr) ν max2922, 2852, 1805, 1625, 1494, 1438, 1419, 1375, 1342,1288, 1230, 1203, 1157, 1114, 1066, 1001, 962, 885, 835, 788, 748, 700cm -1 ;

[0081] HR-ESI-MS m / z 863.3068 [M+H] + (Calculated value C) 52 H 47 O 12 : 863.3062).

[0082] 1H NMR spectrum 1 H NMR and carbon spectroscopy (H NMR) 13 The C NMR data are shown in Table 1.

[0083] 1 H NMR, 13 C NMR, 1 H- 1 The H COSY, HSQC, HMBC, and NOESY NMR spectra are shown in [reference needed]. Figures 1-6 .

[0084] Single crystals of chalcone derivatives were obtained from a mixed solution of ethyl acetate and acetonitrile using a solvent evaporation method. Crystals with good crystal faces and suitable sizes were selected for single-crystal X-ray diffraction analysis using a Rigaku single-crystal diffractometer with Cu Kα target radiation at λ = 1.54184 Å as the light source. The collected diffraction data were analyzed using SHELXTL 6.14 and Olex 21.3.0 analysis software. The single-crystal X-ray diffraction data are shown in Table 2.

[0085] Single-crystal X-ray diffraction structure is shown in Figure 7 .

[0086] Based on the above physicochemical data, NMR data, and single-crystal X-ray diffraction data, it can be seen that the structure of the chalcone derivative prepared in Example 1 is as shown in Formula I, and its configuration is 17R, 17'R and 17S, 17'S.

[0087]

[0088] Table 1. Chalcone derivatives 1 H (400 MHz) and 13 C (100 MHz) NMR data

[0089]

[0090] Table 2 Crystal data of chalcone derivatives

[0091]

[0092]

[0093]

[0094] Example 3: Effects of chalcone derivatives on angiogenesis in HMEC-1 vascular endothelial cells

[0095] 1. Cell culture

[0096] HMEC-1 cells were seeded at a density of 3000 cells per well in 96-well plates and cultured overnight in a humid environment of 37°C and 5% CO2.

[0097] 2. Anti-HMEC-1 cell lumen formation experiment

[0098] HMEC-1 cells suspended in 100 μL of DMEM medium containing different concentrations of the test compounds were seeded into 96-well Matrigel-coated plates. After 6 hours of incubation, tubular structures were observed and photographed using an EVOS XL Core microscope (Thermo Fisher Scientific). The number of branch points was counted using Image-Pro Plus 6.0 software (Media Cybernetics, Rockville, Maryland, USA) to quantitatively analyze lumen formation.

[0099] See results Figure 8 The chalcone derivative prepared in this invention can significantly inhibit angiogenesis in vascular endothelial cells HMEC-1 at low concentrations.

[0100] Example 4: Effects of chalcone derivatives on angiogenesis in zebrafish

[0101] 1. Breeding and rearing of zebrafish embryos

[0102] Male and female transgenic zebrafish strain Tg(fli-1a:EGFP)y1 (characterized by enhanced green fluorescent protein expression in vascular endothelial cells), obtained from the zebrafish screening platform of Guangzhou University of Chinese Medicine, were placed in a breeding box containing E3 embryo culture medium. A transparent plate separated the males and females. After the females laid eggs, the plate was removed, and healthy, normally developing fertilized embryos were collected. The embryos were then cultured in E3 embryo culture medium at 28.5℃. Six hours after fertilization, 0.003% PTU was added to the culture medium to inhibit melanin production, and the culture medium was replaced promptly. Unfertilized or abnormally developing dead embryos were removed.

[0103] 2. Compound intervention

[0104] (1) Solution preparation

[0105] A high-concentration stock solution was prepared by dissolving the chalcone derivative in DMSO. Before use, the stock solution was diluted with E3 embryo culture medium to prepare working solutions of 80 μM and 40 μM for the chalcone derivative.

[0106] (2) Experimental grouping

[0107] The experiment was divided into three groups: a blank control group (containing only E3 embryo culture medium), a compound treatment group (80 μM and 40 μM chalcone derivatives).

[0108] (3) Experimental results

[0109] Twenty-four hours after fertilization, healthy embryos developed to the appropriate stage were selected and randomly assigned to 12-well plates, with 30 embryos placed in each well. The original culture medium was removed, and the prepared chalcone derivative working solution was added to the plate. The culture plate was placed in a constant temperature incubator at 28.5℃ in the dark for 6 hours. After drug administration, the embryos were transferred to E3 culture medium containing 0.02% tricaine solution until the embryos were completely anesthetized. The anesthetized embryos were placed on their sides on a glass slide and fixed with a small amount of 3% methylcellulose.

[0110] Fluorescent images of zebrafish blood vessels were observed and captured using a Leica DMi8 manual fluorescence microscope (Leica Wetzlar GmbH, Germany). Quantitative analysis of intersegmental vessels in each zebrafish embryo was performed using ImageJ software.

[0111] See results Figure 9 The chalcone derivative prepared in this invention can significantly inhibit the formation of new blood vessels in zebrafish.

[0112] The embodiments described above are merely illustrative of several implementations of the present invention, and while the descriptions are relatively specific and detailed, they should not be construed as limiting the scope of the invention patent. It should be noted that those skilled in the art can make various modifications and improvements without departing from the concept of the present invention, and these all fall within the protection scope of the present invention. Therefore, the protection scope of this invention patent should be determined by the appended claims.

Claims

1. A chalcone derivative or a pharmaceutically acceptable salt thereof or a stereoisomer thereof, characterized in that, The molecular structure of the chalcone derivative is shown in Formula I. 。 2. The use of the chalcone derivative of claim 1, or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof, in the preparation of a medicament for inhibiting angiogenesis.

3. The application according to claim 2, characterized in that, The drugs that inhibit angiogenesis are drugs that inhibit tumor angiogenesis, drugs that inhibit angiogenesis in rheumatoid arthritis lesions, drugs that inhibit angiogenesis in diabetic retinopathy lesions, drugs that inhibit angiogenesis in psoriasis lesions, or drugs that inhibit angiogenesis in atherosclerotic lesions.

4. A drug for inhibiting angiogenesis, characterized in that, It is prepared from an active ingredient and pharmaceutically acceptable excipients, wherein the active ingredient comprises the chalcone derivative of claim 1 or a pharmaceutically acceptable salt thereof or a stereoisomer thereof.

5. A method for preparing the chalcone derivative according to claim 1, characterized in that, Includes the following steps: (1) Crush the branches and leaves of the guarful tree, extract them by percolation with an ethanol aqueous solution, concentrate the resulting extract to obtain a crude extract; (2) The crude extract is suspended in water and then extracted with petroleum ether. The resulting extract is concentrated to obtain the petroleum ether extract fraction. (3) The petroleum ether extract is separated and purified to obtain the chalcone derivative.

6. The method for preparing chalcone derivatives according to claim 5, characterized in that, The volume concentration of the ethanol aqueous solution is 90%-95%; And / or, the number of percolation extractions in step (1) is 3-5 times; And / or, the time for each percolation extraction is 40-56 hours; And / or, the ratio of ethanol aqueous solution to guafu wood used in each percolation extraction is 1.6L-2.4L:1kg.

7. The method for preparing chalcone derivatives according to claim 5, characterized in that, The ratio of crude extract to water in step (2) is 1 kg: 3.5 L - 4.5 L; And / or, in step (2), the extraction is performed 4-6 times, and the volume ratio of petroleum ether to water used each time is 0.8-1.2:

1.

8. The method for preparing the chalcone derivative according to any one of claims 5-7, characterized in that, The separation and purification in step (3) includes: sequentially separating and purifying the petroleum ether extract by silica gel column chromatography, reverse ODS column chromatography, Sephadex LH-20 column chromatography, and reverse preparative HPLC.

9. The method for preparing the chalcone derivative according to claim 8, characterized in that, The eluent for the silica gel column chromatography is petroleum ether and ethyl acetate, preferably eluted according to an elution gradient of petroleum ether to ethyl acetate volume ratios of 100:0, 1000:1, 1000:3, 100:5, 100:1, 100:3, 100:5, 100:10, 100:20, 100:50, 100:100, and 0:

100. And / or, the eluent for the reversed ODS column chromatography is methanol and water, preferably eluted according to an elution gradient of methanol to water volume ratio of 40:60, 50:50, 60:40, 70:30, 75:25, 80:20, 85:15, 90:10, 95:5 and 100:

0.

10. The method for preparing the chalcone derivative according to claim 8, characterized in that, The eluent used for separation and purification by the Sephadex LH-20 column is a mixed solvent of dichloromethane and methanol with a volume ratio of 1:0.9-1.1, and the preferred flow rate is 0.8 mL / min-1.2 mL / min. And / or, the conditions for the reverse preparative HPLC separation and purification include: a Cosmosil C18 column, an eluent of acetonitrile and water in a volume ratio of 85-90:15-10, and a flow rate of 2.8 mL / min-3.2 mL / min.

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