Flavonoid derivatives and their preparation method and application

Abstract

The present invention discloses a class of flavonoid derivatives, their preparation methods, and applications. The structural formula is shown in Formula I: wherein X is selected from CH and N, and R1 is selected from H. These compounds can significantly inhibit the activation of STAT3, a cell signaling pathway, at low doses. Experimental results show that these compounds can significantly inhibit the proliferation of various tumor cell lines, including lymphoma, multiple myeloma, cervical cancer, breast cancer, gastric cancer, lung cancer, and pancreatic cancer. They can specifically inhibit STAT3 signaling activation and the expression of the downstream target gene cyclinD1, indicating that these compounds have the potential to be developed as targeted anti-tumor drugs related to the STAT3 signaling pathway.

Description

Technical Field

[0001] The present 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] Numerous studies have demonstrated that overexpression and aberrant activation of signal transducer and activator of transcription 3 (STAT3) are closely associated with the development and poor prognosis of various solid tumors and hematologic malignancies. Studies have found that persistent STAT3 activation is found in approximately 70% of human solid and hematologic 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 as an inactive monomer in the cytoplasm and is subject to a strict negative feedback regulation mechanism. Abnormalities in this negative feedback regulation mechanism or gene mutations can lead to persistently elevated STAT3 phosphorylation levels at tyrosine 705 and serine 727. Activated STAT3 monomers can form homodimers or heterodimers through their SH2 domains and enter the cell nucleus. They bind to specific gene promoter sequences through their DNA binding domains, initiating the transcription and protein expression of downstream genes. These include BCL-2 and BCL-XL, members of the BCL-2 family of proteins associated with mitochondrial apoptosis, and a series of anti-apoptotic factors, such as cyclin D1, which is involved in cell cycle regulation. Furthermore, aberrantly activated STAT3 can promote tumor invasion and metastasis by inducing the expression of matrix metalloproteinases (MMPs, particularly MMP-1, MMP-2, and MMP-9) and other STAT3 target genes. Constitutively activated STAT3 can also promote tumor immune escape by downregulating proinflammatory cytokines / chemokines (essential for antitumor immune responses) and upregulating tumor / immunosuppressive factors (VEGF, IL-10, PD-1 / PD-L1, and COX2, which can inhibit immune responses). Therefore, targeting STAT3 protein is a promising therapeutic strategy for tumor treatment.

[0003] Flavonoids are a class of natural products, ubiquitous in nature, characterized by a wide range of physiological activities and low toxic side effects. In recent years, their anti-tumor effects have garnered extensive attention and research. Research results have shown that flavonoids have significant preventive and therapeutic effects on a variety of common cancers, including lung cancer, breast cancer, colon cancer, prostate cancer, liver cancer, leukemia, ovarian cancer, and gastric cancer. The anti-tumor mechanisms of flavonoids include antioxidant and free radical protection, induction of tumor cell apoptosis, cell cycle effects, immune regulation, inhibition of tumor angiogenesis, inhibition of cyclooxygenase-2, and inhibition of telomerase activity. Although flavonoids have a wide range of anti-cancer effects, their pharmacological activity may be limited due to their poor water solubility. Therefore, improving the pharmacological activity of flavonoid derivatives through chemical modification is of great research significance. 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 objectives, 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, and R1 is selected from H,

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

[0010]

[0011]

[0012] A biologically acceptable salt formed by the above-mentioned flavonoid derivative 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.

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

[0014]

[0015] The specific preparation process is as follows:

[0016] (1) Compound 1 was dissolved in a mixed solution of methanol and saturated ammonium chloride aqueous solution, Fe powder was added at 45-55°C, and the reaction was completed by stirring at this temperature; the reaction solution was filtered through diatomaceous earth, rinsed with ethyl acetate, and the filtrate was extracted with water, the organic phase was collected, and column chromatography was 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; the reaction solution is directly spin-dried and sampled for column chromatography to obtain compound 4;

[0018] (3) Compound 5 and KOH were dissolved in tetrahydrofuran, stirred evenly, and then compound 4 was added. The reaction was completed at room temperature under stirring. The reaction solution was filtered through celite, 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, and the organic phase was dried and subjected to column chromatography to obtain the compound of formula I;

[0020] Preferably, in step (1), the molar ratio of compound 1 and Fe powder is 1:5, and the volume ratio of ethanol and 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 signaling conduction.

[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, and leukemia.

[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 novel flavonoid derivatives, RD001, RD002, RD003, RD004, RD005, RD006, RD007, RD008, RD009, and RD010. These compounds were tested for their anti-tumor cell proliferation effects using CCK assays, and their inhibitory effects on STAT3 signal transduction were confirmed by immunoblotting.

[0029] The results showed 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 summary, the present invention provides a new use of flavonoid derivatives in tumor treatment and its potential molecular mechanism. BRIEF DESCRIPTION OF THE DRAWINGS

[0031] Figure 1 Western blot results of MDA-MB-468 cells 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, the various raw materials used in the reaction can be prepared by those skilled in the art based on 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 modified based on 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 conventional drying method, and the solvent is evaporated by a rotary evaporator under reduced pressure, with the bath temperature not higher than 50°C; the developing solvent and the eluent are both in a volume ratio; (iii) the reaction process is monitored 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 completion. The reaction solution was filtered through diatomaceous earth and washed with ethyl acetate. The filtrate was extracted with water and the organic phase was collected and dried by column chromatography (gradient eluent was a mixed solvent of petroleum ether and ethyl acetate with a volume ratio of 10:1 to 3:1). The collected eluent containing the sample was dried 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 with 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 and stirred evenly. Compound 4 (2.00 g, 5.52 mmol, 1.0 eq) was then added and the mixture was stirred at room temperature until complete reaction. The reaction solution was filtered through diatomaceous earth, washed with ethyl acetate, and the organic phase was subjected to spin-drying column chromatography (eluent: a mixed solvent of dichloromethane and methanol in a volume ratio of 200:1 to 50:1). 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 column chromatography was performed (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 were similar to those of Example 1, except that in step 4, the raw material 7a was replaced by piperazine or piperazine with corresponding substituents.

[0050] Example 2: Inhibitory effects of RD001, RD002, RD003, RD004, RD005, RD006, RD007, RD008, RD009 and RD010 on cell proliferation in breast cancer, liver cancer, pancreatic cancer, gastric cancer, lung cancer, esophageal cancer, cervical cancer, multiple myeloma, diffuse large B-cell lymphoma, etc.

[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. Using 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 microplate reader was read to measure the OD value at an absorption wavelength of 450nm. The results were recorded, and the cell growth curve was drawn with the dose of the compound as the horizontal axis and the absorbance value as the vertical axis. The statistical results of the half-maximal inhibition rate (IC50 value) of the compound on tumor cells are shown in Table 1 below:

[0052] Table 1. CCK-8 assay of the 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, and diffuse large B-cell lymphoma 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 anti-tumor 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 STAT3 phosphorylation 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 seeded into 6-well plates at 2 mL per well. b. Incubate overnight at 37°C in a 5% CO2 incubator. Add various concentrations of RD001 to achieve final concentrations of 0, 100, 300, 1000, and 3000 nM. DMSO was used as a negative control. c. After a further 24 hours of culture, cells were lysed with RIPA buffer and protein was collected.

[0059] II. Cell Harvest and Lysis: a. Discard the supernatant medium and wash the cells twice with pre-chilled PBS. Add 100 μL of pre-chilled RIPA cell lysis buffer (protease inhibitors and PMSF are pre-mixed at a 1:100 ratio) to each well.

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

[0061] for 30 min, vortexing every 6 min.

[0062] c. Centrifuge at 12,000 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 thoroughly for later use; take 80 μL of the remaining cell supernatant, add 20 μL of 5× SDS Loading Buffer, mix thoroughly, and boil in boiling water for 10 minutes. After centrifugation, load 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 A and B required based on the number of standards and samples to be tested. Prepare the working solution at a volume ratio of 50:1 for BCA reagent A to B, vortex and mix thoroughly.

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

[0067]

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

[0069] (4) Remove the 96-well plate and return it to room temperature for 3–5 min. Measure the absorbance of A562 on a microplate reader and save the values ​​in an Excel spreadsheet. Create 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 it to 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 50 μL TEMED 5μL Total 5mL

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

[0076] (6) Remove the protein sample, heat in a 100°C water bath for 5 minutes, and centrifuge at 10,000 rpm for 5 minutes. (7) Fix the gel plate to the electrophoresis tank, add SDS-PAGE electrophoresis buffer, remove the comb, and add the processed protein sample to the sample tank in order, 50 μg of protein per well. (8) Electrophoresis at 80V for 40 minutes. (9) Change the voltage to 120V and electrophoresis for about 1.5 hours until the 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 cotton pad in membrane transfer buffer, clamp it onto the transfer apparatus with tweezers, and place it in the order of blackboard, cotton pad, filter paper, protein gel, PVDF membrane, filter paper, cotton pad and whiteboard, clamp it, and place it on the transfer apparatus. If there are bubbles between each layer, use a glass tube to gently roll them out. (3) Turn on the transfer apparatus and transfer at a constant current of 300mA for 80 minutes. (4) Place the membrane in TBST buffer and rinse 3 times, 8 minutes each time. (5) Block with 20mL of 5% BSA-TBST blocking solution at room temperature for 2 hours. (6) Add primary antibody and incubate at 4℃ 60rpm overnight. (7) Wash the membrane three times with TBST at room temperature on a shaker at 60rpm, 10 minutes each time. (8) Add secondary antibody and incubate at room temperature for 1 hour. (9) Wash the membrane three times with TBST at room temperature, shaking at 60 rpm, for 10 minutes each time. (10) Take 1 mL each of chemiluminescent substrate solution A and solution B and develop the color at room temperature for 2 minutes. (11) Blot the liquid on the membrane with filter paper 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 approximately 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-fold 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 of Tris in 80 mL of water and adjust the pH to 8.8 with 4 N HCl. The volume is then adjusted 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 in 800 mL of distilled water, dissolve it, add 10 mL of 1 mol / L TrisHCl (pH 7.5), adjust the volume 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 of 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 effects and development potential.

[0090] According to the general approach of drug development (conventional anti-tumor in vitro screening first, followed by 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 and not to limit the technical solutions of the present invention. 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 was dissolved in a mixed solution of methanol and saturated ammonium chloride aqueous solution, Fe powder was added at 45-55°C, and the reaction was completed by stirring at this temperature; the reaction solution was filtered through diatomaceous earth, rinsed with ethyl acetate, and the filtrate was extracted with water, the organic phase was collected, and column chromatography was 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, add compound 3, and then react completely at room temperature; the reaction solution is directly spin-dried and sampled for column chromatography to obtain compound 4; (3) Compound 5 and KOH were dissolved in tetrahydrofuran, stirred evenly, and then compound 4 was added. The reaction was completed at room temperature under stirring. The reaction solution was filtered through celite, 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, and the organic phase was dried and subjected to column chromatography to obtain the 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 and Fe powder is 1:5, and the volume ratio of ethanol and 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 biologically acceptable salt thereof according to any one of claims 1 to 3 in the preparation of a medicament, characterized in that: The drug is a drug for treating diseases related to abnormal STAT3 cell signaling conduction.

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 hematological tumors, and the hematological 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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