Flavone derivative as well as preparation method and application thereof

Abstract

The invention discloses a flavone derivative and a preparation method and application thereof, the structural formula of the flavone derivative is shown as a general formula I: # imgabs0, X is selected from CH and N, R1 is selected from H, # imgabs1, # imgabs2 and # imgabs3, and the compound can significantly inhibit activation of a cell signal transduction STAT3 signal under a low dosage. Experimental results show that the compound can significantly inhibit proliferation of cell strains of multiple tumors such as lymphoma, multiple myeloma, cervical cancer, breast cancer, gastric cancer, lung cancer and pancreatic cancer, and can specifically inhibit STAT3 signal activation and expression of a downstream target gene cyclinD1; therefore, the compound has a prospect of being developed into a targeted anti-tumor drug related to the STAT3 signal transduction pathway.

Description

Technical Field

[0001] The invention belongs to the field of tumor targeted therapy, and specifically relates to a flavonoid derivative and a preparation method and application thereof. Background Art

[0002] A large number of studies have shown that the overexpression and abnormal activation of signal transducer and activators of transcriptions 3 (STAT3) are closely related to the occurrence, development and poor prognosis of a variety of solid tumors and hematological tumors. Studies have found that persistent STAT3 activation is found in about 70% of human solid tumors and hematological tumors, including colorectal cancer, lung cancer, melanoma, breast cancer, prostate cancer, kidney cancer, ovarian cancer, liver cancer, pancreatic cancer, multiple myeloma and leukemia. Under normal circumstances, STAT3 exists in the cytoplasm as an inactive monomer and has a strict negative feedback regulation mechanism. When the negative feedback regulation mechanism of STAT3 is abnormal or the gene mutates, it can lead to a continuous increase in the phosphorylation level of STAT3 tyrosine 705 and serine 727 sites. Activated STAT3 monomers can form homodimers or heterodimers through the SH2 domain and enter the cell nucleus, bind to specific gene promoter sequences through the DNA binding domain, and initiate the transcription and protein expression of downstream genes, including BCL-2 and BCL-XL of the BCL-2 protein family related to mitochondrial apoptosis, and the expression of a series of anti-apoptotic factors such as cyclinD1 related to cell cycle regulation. In addition, abnormally activated STAT3 can also promote tumor invasion and metastasis by inducing the expression of matrix metalloproteinases (MMPs, especially MMP-1, MMP-2, MMP-9) and other STAT3 target genes. Constitutively activated STAT3 can also promote tumor immune escape by downregulating proinflammatory cytokines / chemokines (essential for anti-tumor immune response) and upregulating tumor / immunosuppressive factors (VEGF, IL-10, PD1 / PD-L1, COX2 can inhibit immune response). Therefore, targeting STAT3 protein is a potential strategy for tumor treatment.

[0003] Flavonoids are a class of natural products that are ubiquitous in nature. They have a wide range of physiological activities and low toxicity and side effects. In recent years, their anti-tumor effects have received extensive attention and research. Research results show that flavonoids have significant preventive and therapeutic effects on a variety of common cancers such as lung cancer, breast cancer, colon cancer, prostate cancer, liver cancer, leukemia, ovarian cancer, and gastric cancer. The anti-tumor mechanisms of flavonoids are mainly: anti-oxidation and anti-free radicals, inducing tumor cell apoptosis, affecting the cell cycle, regulating immunity, inhibiting tumor angiogenesis, inhibiting cyclooxygenase 2, and inhibiting telomerase activity. Although flavonoids have a wide range of anti-cancer effects, their pharmacological activity may be limited because these compounds are poorly soluble in water. Therefore, it is of great research significance to improve the pharmacological activity of flavonoid derivatives through chemical modification. Summary of the invention

[0004] The purpose of the present invention is to provide a flavonoid derivative and a preparation method and application thereof.

[0005] Based on the above purpose, the present invention adopts the following technical solutions:

[0006] A class of flavonoid derivatives, the structural formula of which is shown in general formula I:

[0007]

[0008] Wherein, X is selected from C, N, R1 is selected from H,

[0009] The above-mentioned flavonoid derivatives are specifically compounds with the following structures:

[0010]

[0011]

[0012] The above-mentioned flavonoid derivatives are biologically acceptable salts formed by at least one of acetic acid, dihydrofolic acid, benzoic acid, citric acid, sorbic acid, propionic acid, oxalic acid, fumaric acid, maleic acid, hydrochloric acid, malic acid, phosphoric acid, sulfurous acid, sulfuric acid, vanillic acid, tartaric acid, ascorbic acid, boric acid, lactic acid and ethylenediaminetetraacetic acid.

[0013] The preparation method of the above-mentioned flavonoid derivatives, the synthetic route is as follows:

[0014]

[0015] The specific preparation process is as follows:

[0016] (1) Compound 1 is dissolved in a mixed solution of methanol and saturated ammonium chloride aqueous solution, Fe powder is added at 45-55° C., and the reaction is stirred at this temperature until the reaction is complete; the reaction solution is filtered through diatomaceous earth, rinsed with ethyl acetate, the filtrate is extracted with water, the organic phase is collected, and the organic phase is spin-dried and column chromatography is performed to obtain compound 2;

[0017] (2) Dissolve triphosgene in dichloromethane, add compound 2 and DIEA at -5 to 5°C, react for 1 hour, add compound 3 and then react completely at room temperature; directly spin dry the reaction solution and perform column chromatography to obtain compound 4;

[0018] (3) Compound 5 and KOH were dissolved in tetrahydrofuran, stirred evenly, and then compound 4 was added, and the reaction was completed at room temperature under stirring; the reaction solution was filtered through diatomaceous earth, washed with ethyl acetate, and the organic phase was spin-dried and column chromatography was performed to obtain compound 6;

[0019] (4) Compound 6, Compound 7 and potassium carbonate were dissolved in DMF and stirred at room temperature until the reaction was complete; the reaction solution was diluted with ethyl acetate and washed with saturated brine, the organic phase was dried and column chromatography was performed to obtain a compound of formula I;

[0020] Preferably, in step (1), the molar ratio of compound 1 to Fe powder is 1:5, and the volume ratio of ethanol to saturated ammonium chloride aqueous solution is 2-3:1; in step (2), the molar ratio of triphosgene, compound 2, DIEA and compound 3 is 1:3:4:3; in step (3), the molar ratio of compound 4, compound 5 and KOH is 1:(1.1-1.5):2.0; in step (4), the molar ratio of compound 6, compound 7 and potassium carbonate is 1:(1.1-1.5):2.

[0021] The use of the above-mentioned flavonoid derivatives and biologically acceptable salts thereof in the preparation of medicines, wherein the medicines are medicines for treating diseases related to abnormal STAT3 cell signal transduction.

[0022] Furthermore, the drug for treating diseases related to abnormal STAT3 cell signaling is a drug for treating hematological tumors such as lymphoma, multiple myeloma, leukemia, etc.

[0023] Furthermore, the drug for treating diseases related to abnormal STAT3 cell signaling is a drug for treating solid tumors such as breast cancer, gastric cancer, colon cancer, esophageal cancer, lung cancer, liver cancer, pancreatic cancer, bladder cancer, cervical cancer, ovarian cancer and prostate cancer.

[0024] Furthermore, the drug for treating diseases related to abnormal STAT3 cell signaling is a drug for treating central nervous system tumors such as retinoblastoma and glioma.

[0025] Furthermore, the drug for treating diseases related to abnormal STAT3 cell signaling is used to treat autoimmune diseases such as psoriasis, rheumatoid arthritis and pulmonary fibrosis.

[0026] Another object of the present invention is to provide a class of small molecule compounds with targeted anti-tumor activity.

[0027] The tumor may specifically be a tumor with high expression or constitutive activation of STAT3, including but not limited to lymphoma, multiple myeloma, cervical cancer, breast cancer, gastric cancer, lung cancer and pancreatic cancer.

[0028] Specifically, the present invention synthesized a class of flavonoid derivatives RD001, RD002, RD003, RD004, RD005, RD006, RD007, RD008, RD009 and RD010 with a new structure. The proliferation inhibition effect of these compounds on tumor cells was detected by CCK method, and their inhibitory effect on STAT3 signal transduction was confirmed by immunoblotting method.

[0029] The results show that the compounds RD001, RD002, RD003, RD004, RD005, RD006, RD007, RD008, RD009 and RD010 of the present invention can effectively inhibit the proliferation of lymphoma, multiple myeloma, cervical cancer, breast cancer, gastric cancer, lung cancer and pancreatic cancer cells.

[0030] In conclusion, the present invention provides a new use of flavonoid derivatives in tumor treatment and potential molecular mechanism. BRIEF DESCRIPTION OF THE DRAWINGS

[0031] Figure 1 This is the Western blot result after MDA-MB-468 cells were treated with RD001 (0, 100, 300, 1000 and 3000 nM) for 24 h. DETAILED DESCRIPTION

[0032] In order to make the technical purpose, technical solution and beneficial effects of the present invention more clear, the technical solution of the present invention is further described below in conjunction with the accompanying drawings and specific embodiments.

[0033] In the method for synthesizing the compound of formula I of the present invention, various raw materials used in the reaction can be prepared by those skilled in the art according to prior knowledge, or can be prepared by methods known in the literature, or can be purchased commercially. The intermediates, raw materials, reagents, reaction conditions, etc. used in the above reaction schemes can be appropriately changed according to the prior knowledge of those skilled in the art.

[0034] In the present invention, unless otherwise specified, (i) the temperature is expressed in degrees Celsius (°C), and the operation is carried out at room temperature; more specifically, the room temperature refers to 20-30°C; (ii) the organic solvent is dried by a common drying method, and the solvent is evaporated by a rotary evaporator under reduced pressure, and the bath temperature is not higher than 50°C; the developing agent and the eluent are both in volume ratio; (iii) the reaction process is tracked by thin layer chromatography (TLC); (iv) the final product has a satisfactory proton nuclear magnetic resonance ( 1 H-NMR).

[0035] Example 1: Synthesis of RD001 compound

[0036]

[0037] The name of compound RD001 is 1-((4-oxo-4H-chromen-7-yl)methyl)-3-(4-(4-(3-(piperidin-1-yl)propoxy)phenoxy)naphthalen-1-yl)urea

[0038] Its synthetic route is as follows:

[0039]

[0040] Step 1. 4-fluoronaphthalen-1-amine (Intermediate 2)

[0041] Compound 1 (5 g, 26.16 mmol, 1.0 eq) was dissolved in a mixed solution of 50 mL of methanol and 20 mL of saturated ammonium chloride aqueous solution, and Fe powder (7.3 g, 130.78 mmol, 5.0 eq) was added at 50° C., and the reaction was stirred at this temperature until complete; the reaction solution was filtered through diatomaceous earth, rinsed with ethyl acetate, and the filtrate was extracted with water, and the organic phase was collected and dried by column chromatography (the gradient eluent was a mixed solvent of petroleum ether and ethyl acetate in a volume ratio of 10:1 to 3:1), and the collected eluent containing the sample was dried by spin drying to obtain compound 2;

[0042] Step 2. 1-(4-fluoronaphthalen-1-yl)-3-((4-oxo-4H-chromen-7-yl)methyl)urea (Intermediate 4)

[0043] Triphosgene (2 g, 6.74 mmol, 1.0 eq) was dissolved in 30 mL of dichloromethane, and compound 2 (3.26 g, 20.22 mmol, 3.0 eq) and DIEA (3.48 g, 26.96 mmol, 4.0 eq) were added at -5 to 5°C. After reacting for 1 hour, compound 3 (3.54 g, 20.22 mmol, 3.0 eq) was added and the reaction was completed at room temperature; the reaction solution was directly spin-dried and mixed with a sample column chromatography (the eluent was a mixed solvent of dichloromethane and methanol in a volume ratio of 200:1 to 70:1), and the collected eluent containing the sample was spin-dried to obtain compound 4.

[0044] Step 3. 1-(4-(4-(2-bromoethoxy)phenoxy)naphthalen-1-yl)-3-((4-oxo-4H-chromen-7-yl)methyl)urea (Intermediate 6)

[0045] Compound 5 (1.32 g, 6.07 mmol, 1.1 eq) and KOH (0.62 g, 11.04 mmol, 2.0 eq) were dissolved in 30 mL of tetrahydrofuran, stirred evenly, and then compound 4 (2.00 g, 5.52 mmol, 1.0 eq) was added, and the reaction was completed at room temperature under stirring; the reaction solution was filtered through diatomaceous earth, washed with ethyl acetate, and the organic phase was spin-dried for column chromatography (the eluent was a mixed solvent of dichloromethane and methanol in a volume ratio of 200:1 to 50:1), and the collected eluent containing the sample was spin-dried to obtain compound 6;

[0046] Step 4.

[0047] Compound 6 (1.00 g, 1.79 mmol, 1.0 eq), compound 7a (0.17 g, 1.97 mmol, 1.1 eq) and potassium carbonate (0.49 g, 3.58 mmol, 2 eq) were dissolved in 30 mL of DMF and stirred at room temperature until the reaction was complete; the reaction solution was diluted with ethyl acetate and washed with saturated brine, the organic phase was dried and spin-dried for column chromatography (the eluent was a mixed solvent of dichloromethane and methanol in a volume ratio of 100:1 to 10:1), and the collected eluent containing the sample was spin-dried to obtain RD001.

[0048] RD001 NMR analysis, 1H NMR (CDCl3, 300MHz) δ: 8.90 (s, 1H), 8.3 (d, J = 7.5Hz, 1H), 8.06 (d, J = 7.4Hz, 1H), 7.71 (d, J = 7.4Hz, 1H), 7.63 (t, J = 7.4Hz, 1H) , 7.46 (t, J = 7.4Hz, 1H), 7.32 (t, J = 7.4Hz, 1H), 7.2 (d, J = 7.4Hz, 1H), 7.01 (s, 1H), 6.95 (t, J = 7.5Hz, 2H), 6.94 (d, J = 7.5Hz, 1H) , 6.92 (d, J = 7.5Hz, 1H), 6.91 (d, J = 7.5Hz, 1H), 6.35 (d, J = 7.5Hz, 1H), 6.29 (d, J = 7.5Hz, 1H), 4.25 (d, J = 7.5Hz, 1H), 4.05 (t, J = 7.5Hz, 2H), 2.48 (t, J=7.5Hz, 2H), 2.42 (m, J=7.5Hz, 4H), 1.83 (m, J=7.5Hz, 2H), 1.49 (m, J=7.5Hz, 4H), 1.37 (m, J=7.5Hz, 2H).

[0049] The synthesis methods of RD002, RD003, RD004, RD005, RD006, RD007, RD008, RD009 and RD010 are similar to those of Example 1, except that in step 4, the raw material 7a is replaced by piperazine or piperazine with corresponding substituents.

[0050] Example 2: Proliferation inhibition of RD001, RD002, RD003, RD004, RD005, RD006, RD007, RD008, RD009 and RD010 on breast cancer, liver cancer, pancreatic cancer, gastric cancer, lung cancer, esophageal cancer, cervical cancer, multiple myeloma, diffuse large B-cell lymphoma and other cells

[0051] MDA-MB-468, HepG2, BxPC-3, MGC803, H460, KYSE450, Hela, MM.1S, and OCI-LY3 cells in the logarithmic growth phase were collected and counted, and the cell suspension concentration was adjusted to 5×10 4 / mL, added to a 96-well cell culture plate, with a volume of 100μL per well. Taking DMSO as a solvent control, the compounds RD001, RD002, RD003, RD004, RD005, RD006, RD007, RD008, RD009 and RD010 described in the present invention were diluted with DMSO and added to the culture wells, so that the final concentrations of the compounds in the system were 0.001, 0.003, 0.01, 0.03, 0.1, 0.3, 1, 3 and 10 (μmol / L), respectively. After continuing to culture for 72h, 10 / 20μL of CCK-8 solvent was added to each well, incubated at 37°C for 3h, and the OD value at an absorption wavelength of 450nm was measured by an enzyme reader. The results were recorded, and the cell growth curve was drawn with the dosage of the compound as the horizontal axis and the absorbance value as the vertical axis. The statistical results of the half inhibition rate (IC50 value) of the compound on tumor cells are shown in Table 1 below:

[0052] Table 1. CCK-8 assay of the proliferation inhibitory effects of RD001, RD002, RD003, RD004, RD005, RD006, RD007, RD008, RD009 and RD010 on breast cancer, liver cancer, pancreatic cancer, gastric cancer, lung cancer, esophageal cancer, cervical cancer, multiple myeloma, diffuse large B-cell lymphoma and other cells.

[0053]

[0054]

[0055]

[0056] The table shows that RD001, RD002, RD003, RD004, RD005, RD006, RD007, RD008, RD009 and RD010 have good proliferation inhibitory effects on breast cancer, liver cancer, pancreatic cancer, gastric cancer, lung cancer, esophageal cancer, cervical cancer, multiple myeloma, diffuse large B-cell lymphoma and other cells, especially in breast cancer, multiple myeloma and diffuse large B-cell lymphoma cells. The tumor inhibitory activity is stronger. This application uses RD001 as an example to conduct a preliminary study on the anti-tumor mechanism of this type of compound.

[0057] Example 3: Effect of RD001 on the phosphorylation of STAT3 in MDA-MB-468 cells

[0058] 1. Cell culture and drug addition: a. Take MDA-MB-468 cells in the logarithmic growth phase and adjust the density to 2×10 5A single cell suspension of 100 cells / mL was inoculated into a 6-well plate with 2 mL of cell suspension per well. b. Incubate overnight in a 37°C, 5% CO2 incubator, add different concentrations of RD001 to a final concentration of 0, 100, 300, 1000, 3000 nM), and DMSO as a negative control. c. After 24 hours of continuous culture, cells were lysed with RIPA lysis buffer and proteins were collected.

[0059] 2. Cell collection and lysis: a. Discard the upper culture medium and wash the cells twice with pre-cooled PBS. Add 100 μL of pre-cooled RIPA cell lysis buffer (protease inhibitors and PMSF are added to the lysis buffer in a ratio of 1:100 and mixed well) to each well.

[0060] b. Lyse on ice for 3 minutes, scrape the cells with a cell scraper and collect them into a 1.5mL EP tube; place on ice for lysis

[0061] 30 min, vortexing every 6 min.

[0062] c. Centrifuge at 12000 g for 10 min at 4°C and transfer the cell supernatant to a new EP tube.

[0063] d. Divide the cell supernatant into two parts: take 5 μL and add it to a 1.5 mL EP tube for BCA protein content measurement, then add 45 μL of 1× PBS and mix well for later use; take 80 μL of the remaining cell supernatant, add 20 μL of 5× SDS Loading Buffer, mix well, boil in boiling water for 10 min, centrifuge and load on the sample or store in a -20°C refrigerator.

[0064] e. Protein concentration determination steps:

[0065] (1) Preparation of BCA working solution: Calculate the total amount of mixed working solution of A and B required based on the number of standards and samples to be tested. Prepare the working solution in a volume ratio of 50:1 for BCA solids A and B, vortex and mix well for later use.

[0066] (2) 1× PBS diluted protein standard:

[0067]

[0068] (3) Take 25 μL of each of the protein standard solution and the sample supernatant diluted with PBS (10-fold dilution) and add them to a new 96-well plate. Then add 200 μL of the pre-prepared BCA working solution and mix thoroughly. Be careful not to blow and create bubbles. Cover the 96-well plate tightly and react in a 37°C incubator for 30 minutes.

[0069] (4) Take out the 96-well plate and return it to room temperature for 3-5 minutes. Measure the absorbance value of A562 on an ELISA reader and copy the obtained value to an Excel spreadsheet. Make a standard curve and calculate the protein content of 1 μL of each sample for protein loading.

[0070] 3. SDS-PAGE: (1) Fix the gel plate and prepare 10% SDS-PAGE separation gel.

[0071] Prepare separation gel according to the following table: 10 mL

[0072] Deionized water 4.0mL 30%(m / v)Acrylamide 3.3mL 1.5M Tris-HCl (pH 8.8) buffer 2.5mL 10%(m / v)SDS 0.1mL 10%(m / v)APS 0.1mL TEMED 4μL Total 10mL

[0073] (2) Add the mixed separation gel to two gel plates, add to the position 1.0 cm from the top, fill the gel plates with anhydrous ethanol, and let it stand for 30 to 45 minutes. (3) After the separation gel is solidified, pour out the remaining anhydrous ethanol and absorb the remaining anhydrous ethanol with filter paper. (4) Prepare 5 mL of 5% concentrated gel according to the table below

[0074] Deionized water 2.77mL 30%(m / v)Acrylamide 830μL 0.5M Tris-HCl (pH 6.8) buffer 1.26mL 10%(m / v)SDS 50μL 10%(m / v)APS 50μL TEMED 5μL Total 5mL

[0075] (5) Slowly add the prepared concentrated glue to the glue plate to avoid generating bubbles, insert a comb, and let it stand for 30 to 45 minutes.

[0076] (6) Take out the protein sample, heat it in a water bath at 100°C for 5 minutes, and centrifuge it at 10,000 rpm for 5 minutes. (7) Fix the gel plate to the electrophoresis tank, add SDS-PAGE electrophoresis buffer, pull out the comb, and add the processed protein samples to the sample tank in order, 50 μg of protein per well. (8) Perform electrophoresis at 80V for 40 minutes. (9) Change the voltage to 120V for about 1.5 hours until bromophenol blue runs out of the colloid;

[0077] IV. Western-blot: (1) Rinse the SDS-PAGE gel after electrophoresis in TBST buffer once, and soak the protein gel in transfer buffer. (2) Soak a layer of cotton pad in membrane transfer buffer, clamp it to the transfer instrument with tweezers, and place it in the order of blackboard, cotton pad, filter paper, protein gel, PVDF membrane, filter paper, cotton pad and white board, clamp it, and put it into the transfer instrument. If there are bubbles between each layer, use a glass tube to gently roll them out. (3) Turn on the transfer instrument and transfer at 300mA constant current for 80min. (4) Put the membrane in TBST buffer and rinse 3 times, 8min each time. (5) Block with 20mL of 5% BSA-TBST blocking solution at room temperature for 2h. (6) Add primary antibody and incubate overnight at 4℃60rpm. (7) Wash the membrane three times with TBST at room temperature, shaking at 60rpm, 10min each time. (8) Add secondary antibody and incubate at room temperature for 1h. (9) Wash the membrane three times with TBST at room temperature, shaking at 60 rpm, for 10 min each time. (10) Take 1 mL each of chemiluminescent substrate solution A and solution B, and color develop at room temperature for 2 min. (11) Use filter paper to absorb the liquid on the membrane and expose it to light.

[0078] 5. Reagent preparation:

[0079] (1) 10% SDS: Weigh 1 g of high-purity (electrophoresis grade) SDS into a 10 mL centrifuge tube, add about 8 mL of deionized water, heat to dissolve, dilute to 10 mL, and store at room temperature.

[0080] (2) 10% ammonium persulfate (AP): weigh 1 g of ammonium persulfate, add about 10 mL of deionized water, stir to dissolve, and store at 4°C.

[0081] (3) 5× electrophoresis buffer: weigh 15.1 g of Tris, 94 g of Glycine, and 5.0 g of SDS into a beaker, add 1 L of double distilled water to dissolve, store at room temperature, and dilute 5 times before use.

[0082] (4) Transfer buffer: Weigh 5.8 g of Tris, 11.6 g of glycine, and 0.75 g of SDS into a beaker, add 700 mL of double distilled water, dissolve and adjust the volume to 800 mL, and finally add 200 mL of methanol.

[0083] (5) 1.5 mol / L Tris-HCl, 100 mL: Dissolve 18.15 g Tris in 80 mL water and adjust the pH to 8.8 with 4 N HCl. Make up to 100 mL.

[0084] (6) 0.5 mol / L Tris-HCl, 1000 mL: Weigh 60.5 g of Tris base, add water to 850 mL, add concentrated hydrochloric acid and stir until completely dissolved, then adjust the pH to 6.8 and add water to 1 L.

[0085] (7) TBS buffer: Weigh 8.8 g of NaCl and dissolve it in 800 mL of distilled water. Add 10 mL of 1 mol / L TrisHCl (pH 7.5) to make up to 1 L and store at room temperature.

[0086] (8) TBST buffer: Add 500 μL of 20% Tween 20 to 1 L of TBS buffer to make the final concentration of Tween 20 0.1%. Prepare it before use.

[0087] (9) Blocking solution, antibody diluent: Add 5% skim milk powder or BSA to TBST buffer and prepare it before use.

[0088] like Figure 1 The results showed that treatment with 1000nM and 3000nM RD001 could effectively downregulate the expression levels of p-STAT3(Y705), p-STAT3(S727) and STAT3's downstream target protein CyclinD1.

[0089] The above results show that RD001, RD002, RD003, RD004, RD005, RD006, RD007, RD008, RD009 and RD010 can significantly inhibit the proliferation of breast cancer, liver cancer, pancreatic cancer, gastric cancer, lung cancer, esophageal cancer, cervical cancer, multiple myeloma, diffuse large B-cell lymphoma and other cells, and can effectively downregulate the expression levels of p-STAT3 (Y705), p-STAT3 (S727) and STAT3's downstream target protein CyclinD1. Therefore, this type of drug has good anti-cancer effect and development potential.

[0090] According to the general approach of drug development (conventional anti-tumor in vitro screening first, then targeted research), the compounds of the present invention can be applied to cancer treatment drugs related to abnormal cell proliferation, and can be prepared as anti-tumor drugs and autoimmune disease drugs by mixing with human-acceptable salts or with pharmaceutical carriers.

[0091] Finally, it should be noted that the above embodiments are only used to illustrate but not limit the technical solutions of the present invention, and any equivalent substitutions of the present invention and any modifications or partial substitutions that do not depart from the spirit and scope of the present invention should be included in the scope of protection of the claims of the present invention.

Claims

1. A flavonoid derivative, characterized in that: The structural formula is shown in general formula I: Wherein, X is selected from CH, N, R1 is selected from H, 2. The flavonoid derivative according to claim 1, characterized in that Specifically, the compound has the following structure:

3. A biologically acceptable salt of the flavonoid derivative according to claim 1 or 2 and at least one of acetic acid, dihydrofolic acid, benzoic acid, citric acid, sorbic acid, propionic acid, oxalic acid, fumaric acid, maleic acid, hydrochloric acid, malic acid, phosphoric acid, sulfurous acid, sulfuric acid, vanillic acid, tartaric acid, ascorbic acid, boric acid, lactic acid and ethylenediaminetetraacetic acid.

4. The method for preparing the flavonoid derivative according to claim 1 or 2, characterized in that: The synthetic route is as follows: The specific preparation process is as follows: (1) Compound 1 is dissolved in a mixed solution of methanol and saturated ammonium chloride aqueous solution, Fe powder is added at 45-55° C., and the reaction is stirred at this temperature until the reaction is complete; the reaction solution is filtered through diatomaceous earth, rinsed with ethyl acetate, the filtrate is extracted with water, the organic phase is collected, and the organic phase is spin-dried and column chromatography is performed to obtain compound 2; (2) Dissolve triphosgene in dichloromethane, add compound 2 and DIEA at -5 to 5°C, react for 0.5 to 1.5 hours, then add compound 3 and react completely at room temperature; directly spin dry the reaction solution and perform column chromatography to obtain compound 4; (3) Compound 5 and KOH were dissolved in tetrahydrofuran, stirred evenly, and then compound 4 was added, and the reaction was completed at room temperature under stirring; the reaction solution was filtered through diatomaceous earth, washed with ethyl acetate, and the organic phase was spin-dried and column chromatography was performed to obtain compound 6; (4) Compound 6, Compound 7 and potassium carbonate were dissolved in DMF and stirred at room temperature until the reaction was complete; the reaction solution was diluted with ethyl acetate and washed with saturated brine, the organic phase was dried and column chromatography was performed to obtain a compound of formula I.

5. The method for preparing the flavonoid derivative according to claim 4, characterized in that: In step (1), the molar ratio of compound 1 to Fe powder is 1:5, and the volume ratio of ethanol to saturated ammonium chloride aqueous solution is 2-3:1; in step (2), the molar ratio of triphosgene, compound 2, DIEA and compound 3 is 1:3:4:3; in step (3), the molar ratio of compound 4, compound 5 and KOH is 1:(1.1-1.5):2.0; in step (4), the molar ratio of compound 6, compound 7 and potassium carbonate is 1:(1.1-1.5):

2.

6. Use of the flavonoid derivative and its biologically acceptable salt according to any one of claims 1 to 3 in the preparation of a medicament, characterized in that: The medicine is a medicine for treating diseases related to abnormal STAT3 cell signal transduction.

7. The use according to claim 6, characterized in that The drug for treating diseases related to abnormal STAT3 cell signaling is a drug for treating blood system tumors, and the blood system tumors refer to lymphoma, multiple myeloma or leukemia.

8. The use according to claim 6, characterized in that The drug for treating diseases related to abnormal STAT3 cell signaling is a drug for treating solid tumors, and the solid tumors refer to breast cancer, gastric cancer, colon cancer, esophageal cancer, lung cancer, liver cancer, pancreatic cancer, bladder cancer, cervical cancer, ovarian cancer and prostate cancer.

9. The use according to claim 6, characterized in that: The drug for treating diseases related to abnormal STAT3 cell signaling is a drug for treating central nervous system tumors, and the central nervous system tumors refer to retinoblastoma and glioma.

10. The use according to claim 6, characterized in that The drug for treating diseases related to abnormal STAT3 cell signaling is a drug for treating autoimmune diseases, and the autoimmune diseases refer to psoriasis, rheumatoid arthritis and pulmonary fibrosis.

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