A class of piperazineamide derivatives and their preparation method and application

Abstract

The present invention discloses a class of piperazinamide derivatives, their preparation methods, and applications, belonging to the field of targeted tumor therapeutics. The structural formula is shown in General Formula I: #imgabs0#, wherein n=1, 2, 3, 4, 5, ..., 10, and R1 is selected from #imgabs1##imgabs2#. These compounds can significantly inhibit the activation of the STAT3 signaling pathway in cell signaling at low doses. Experimental results show that these compounds can significantly inhibit the proliferation of various tumor cell lines, including lymphoma, multiple myeloma, leukemia, breast cancer, gastric cancer, lung cancer, and pancreatic cancer. They can specifically inhibit STAT3 signaling activation and the expression of the downstream target gene C-myc, 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 therapeutic drugs, and specifically relates to a class of piperazine amide derivatives 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 and hematologic tumors. Under normal circumstances, STAT3 exists as an inactive monomer in the cytoplasm and is subject to a strict negative feedback regulatory mechanism. Abnormalities in this negative feedback regulatory mechanism or gene mutations can lead to persistently elevated phosphorylation levels at STAT3 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 transcription and protein expression of downstream genes, including BCL-2 and BCL-XL, members of the BCL-2 family of proteins involved in mitochondrial apoptosis, and a series of anti-apoptotic factors, such as C-myc, involved in cell cycle regulation. In animal xenograft models or in vitro cultured tumor cell lines with persistent STAT3 activation, knocking down STAT3 protein or inhibiting STAT3 activation with small molecule compounds can effectively inhibit tumor cell growth, induce apoptosis, and reduce tumor metastasis. STAT3 has become a popular target for cancer treatment. Although several STAT3 inhibitors, such as BBI608, TTI101, and WP1066, have entered clinical trials in cancer patients both domestically and internationally, no inhibitors directly targeting STAT3 have successfully reached market.

[0003] To develop STAT3-targeted anti-tumor drugs, we recently synthesized a novel class of naphthylurea-piperazine compounds. Through biological analysis, we found that these compounds significantly inhibited STAT3 signaling and proliferation in various tumor cells, demonstrating promising targeted anti-tumor activity.

[0004] The present invention aims to reveal the anti-tumor effects and potential pharmacological mechanisms of a new class of piperazinamide derivatives, as well as the potential applications of such compounds in the clinical treatment of lymphoma, multiple myeloma, leukemia, breast cancer, gastric cancer, lung cancer and pancreatic cancer. Summary of the Invention

[0005] The purpose of the present invention is to provide a class of piperazine amide derivatives and their preparation method and application.

[0006] Based on the above objectives, the present invention adopts the following technical solutions:

[0007] A class of piperazine amide derivatives, the structural formula of which is shown in general formula I:

[0008]

[0009] Formula I, wherein n=1, 2, 3, 4, 5...10, R1 is selected from

[0010] The above-mentioned piperazine amide derivative is specifically a compound with the following structure:

[0011]

[0012] A biologically acceptable salt formed by the above-mentioned piperazinamide 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 piperazine amide derivatives and the synthetic route are as follows:

[0014]

[0015] The specific preparation process is as follows:

[0016] (1) Compound 1 and t-BuOK were dissolved in tetrahydrofuran, stirred evenly, and then compound 2 was added. The mixture was stirred and reacted at room temperature until complete. The reaction solution was filtered through diatomaceous earth, washed with ethyl acetate, and the organic phase was subjected to spin column chromatography to obtain compound 3.

[0017] (2) Compound 3, compound 4 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 compound 5;

[0018] (3) Compound 5 was dissolved in a mixed solution of ethanol 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 dried by column chromatography to obtain compound 6;

[0019] (4) Dissolve triphosgene in dichloromethane, add compound 6 and DIEA at -5 to 5°C, react for 0.5 to 1 hour, add compound 7, and stir at room temperature until the reaction is complete; the reaction solution is directly spin-dried and subjected to column chromatography to obtain the compound of formula I.

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

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

[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, 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, leukemia, breast cancer, gastric cancer, lung cancer and pancreatic cancer.

[0028] Specifically, the present invention synthesized a novel class of piperazinamide compounds, RD240401, RD240402, RD240403, RD240404, RD240405, RD240406, RD240407, RD240408, RD240409, RD240410, and RD240411. These compounds were tested for their inhibitory effects on tumor cell proliferation using the CCK assay, and their inhibitory effects on STAT3 signaling were confirmed by immunoblotting.

[0029] The results showed that the compounds RD240401, RD240402, RD240403, RD240404, RD240405, RD240406, RD240407, RD240408, RD240409, RD240410 and RD240411 of the present invention can effectively inhibit the proliferation of lymphoma, multiple myeloma, leukemia, breast cancer, gastric cancer, lung cancer and pancreatic cancer cells.

[0030] In summary, the present invention provides a new piperazinamide compound and its derivatives for use in tumor treatment and potential molecular mechanisms. BRIEF DESCRIPTION OF THE DRAWINGS

[0031] Figure 1 Western blot results of lymphoma MDA-MB-468 cells treated with RD240404 (0, 10, 30, 100 and 300 nM) for 24 hours. 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 RD240401 compound

[0036]

[0037] The name of compound RD240401 is 1-(4-((4-(2-(4-(2-(1H-indol-1-yl)acetyl)piperazin-1-yl)ethoxy)benzyl)oxy)naphthalen-1-yl)-3-(pyridin-4-ylmethyl)urea,

[0038] Its synthetic route is as follows:

[0039]

[0040] Step 1. 1-((4-(2-bromoethoxy)benzyl)oxy)-4-nitronaphthalene (Intermediate 3)

[0041] Raw material 1 (2 g, 8.65 mmol, 1.0 eq) and t-BuOK (0.97 g, 8.65 mmol, 1.0 eq) were dissolved in 20 ml of tetrahydrofuran and stirred for 10 minutes. Raw material 2 (1.65 g, 8.65 mmol, 1.0 eq) was then added and stirred at room temperature for 2 hours. TLC (PE / EA = 4 / 1, Rf / product = 0.25) showed that the raw material reaction was complete and new spots were generated. The reaction solution was filtered through celite and washed with ethyl acetate. The organic phase was dried and passed through a column and eluted with (PE / EA = 10 / 1 to 1 / 1) to obtain intermediate 3 (2.20 g, 63.2%) as a yellow solid.

[0042] Step 2. 2-(1H-indol-1-yl)-1-(4-(2-(4-(((4-nitronaphthalen-1-yl)oxy)methyl)phenoxy)ethyl)piperazin-1-yl)ethan-1-one (Intermediate 5)

[0043] Intermediate 3 (1 g, 2.49 mmol, 1.0 eq), starting material 4 (0.5 g, 2.49 mmol, 1.0 eq) and potassium carbonate (0.34 g, 2.49 mmol, 1.0 eq) were dissolved in 20 mL of DMF and stirred at room temperature for 12 hours. TLC (DCM / MeOH = 20 / 1, Rf / product = 0.35) showed that the starting material reaction was complete and new spots were generated. The reaction solution was diluted with 100 mL of ethyl acetate and washed three times with saturated brine (100 mL * 3). The organic phase was dried, spin-dried, mixed, and passed through a column, which was eluted with (DCM / MeOH = 100 / 1 to 20 / 1) to obtain intermediate 5 (750 mg, 57.2%) as a yellow solid.

[0044] Step 3. 1-(4-(2-(4-(((4-aminonaphthalen-1-yl)oxy)methyl)phenoxy)ethyl)piperazin-1-yl)-2-(1H-indol-1-yl)ethan-1-one (Intermediate 6)

[0045] Intermediate 5 (630 mg, 1.14 mmol, 1.0 eq) was dissolved in 20 ml of ethanol and 10 ml of saturated aqueous ammonium chloride solution. Fe powder (0.32 g, 5.71 mmol, 5.0 eq) was added at 50°C, and the reaction was stirred at 50°C for 1 hour. TLC (DCM / MeOH = 20 / 1, Rf / product = 0.25) showed that the reaction of the raw materials was complete and new spots were generated. The reaction solution was filtered through celite and washed with ethyl acetate. The filtrate was extracted with water and the organic phase was collected, dried, and passed through a column and washed with (DCM / MeOH = 100 / 1 to 20 / 1) to obtain intermediate 6 (400 mg, 70.7%) as a yellow solid.

[0046] Step 4.1-(4-((4-(2-(4-(2-(1H-indol-1-yl)acetyl)piperazin-1-yl)

[0047] ethoxy)benzyl)oxy)naphthalene-1-yl)-3-(pyridin-4-ylmethyl)urea(RD240401)

[0048] Triphosgene (100 mg, 0.37 mmol, 1.0 eq) was dissolved in 20 ml of dichloromethane. Intermediate 6 (500 mg, 1.01 mmol, 3.0 eq) and DIEA (130 mg, 1.01 mmol, 3.0 eq) were added at 0°C. After stirring for 1 hour, starting material 7 (109 mg, 1.01 mmol, 3.0 eq) was added and stirred at room temperature overnight. TLC (DCM / MeOH = 10 / 1, Rf / product = 0.2) showed that the starting material had reacted completely and a new spot was generated. The reaction solution was directly spin-dried and mixed, then passed through a column and eluted with (DCM / MeOH = 50 / 1 to 10 / 1) to obtain RD240401 (215 mg, 33.8%) as a brown solid.

[0049] RD240401 NMR analysis, 1H NMR(DMSO-d6,300MHz)δ:8.32(s,1H),8.19(d,J=8.0Hz,2H),8.01(d,J=8.0Hz,2H),7.68(d,J=8.0Hz,2H),7.58-7.26(m,8H),7.11-7.08( m,5H),7.05-6.98(m,3H),6.82(m,1H),5.20(s,2H),4.34(d,J=4.0Hz,2H),4.11(d,J=4.0Hz,4H),2.52(m,2H),1.53(m,4H),1.40(m,2H).

[0050] The synthesis methods of RD240402, RD240403, RD240404, RD240405, RD240406, RD240407, RD240408, RD240409, RD240410 and RD240411 were similar to those of Example 1, except that in step 2, the raw material 4a was replaced with piperazine having the corresponding substituent.

[0051] Example 2: Inhibitory Effects of RD240401, RD240402, RD240403, RD240404, RD240405, RD240406, RD240407, RD240408, RD240409, RD240410, and RD240411 on Proliferation of Lymphoma, Multiple Myeloma, Leukemia, Breast Cancer, Gastric Cancer, Lung Cancer, and Pancreatic Cancer Cells

[0052] Tumor cells in the logarithmic growth phase were collected and the cell suspension concentration was adjusted to 5×10 4 / mL, added to a 96-well cell culture plate with a volume of 100 μL per well. Using DMSO as a solvent control, the novel naphthyl urea-piperidine compounds RD240401, RD240402, RD240403, RD240404, RD240405, RD240406, RD240407, RD240408, RD240409, RD240410, and RD240411 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 48 hours of culture, 10 μL of CCK-8 solution was added to each well and incubated at 37°C for 3 hours. The OD value at an absorption wavelength of 450 nm was measured using a microplate reader and the results were recorded. A cell growth curve was plotted with the compound dose as the abscissa and the absorbance as the ordinate. The statistical results of the half-maximal inhibition rate (IC50 value) of RD240401, RD240402, RD240403, RD240404, RD240405, RD240406, RD240407, RD240408, RD240409, RD240410, and RD240411 on tumor cells are shown in Table 1.

[0053] Table 1:

[0054]

[0055]

[0056]

[0057]

[0058] The results in Table 1 show that RD240401, RD240402, RD240403, RD240404, RD240405, RD240406, RD240407, RD240408, RD240409, RD240410 and RD240411 have good proliferation inhibitory effects on tumor cells such as lymphoma, multiple myeloma, leukemia, gastric cancer, lung cancer and pancreatic cancer. In particular, RD240404 has the strongest tumor inhibitory activity on lymphoma MDA-MB-468. This application focuses on further research on the anti-tumor effect of this compound.

[0059] Example 3: Inhibitory effect of RD240404 on STAT3 phosphorylation and C-myc and CyclinD1 expression in MDA-MB-468 cells

[0060] 1. Cell culture and drug addition: a. Take MDA-MB-468 cells in the logarithmic growth phase and adjust the density to 2×10 5 A 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 humidified incubator. Add RD240404 at various concentrations (final concentrations: 0, 10, 30, 100, and 300 nM) and 300 nM RDf001 as controls. c. After a further 24 hours of incubation, cells were lysed with RIPA buffer and protein was collected.

[0061] 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.

[0062] b. Lyse on ice for 3 minutes. Scrape the cells with a cell scraper and collect them into a 1.5mL Eppendorf tube. Lyse on ice for 30 minutes, vortexing every 6 minutes. c. Centrifuge at 12,000g for 10 minutes at 4°C. d. Transfer the cell supernatant to a new Eppendorf tube. d. Divide the cell supernatant into two parts: 1. Transfer 5μL of the cell supernatant to a 1.5mL Eppendorf tube for BCA protein analysis, then add 45μL of 1× PBS and mix thoroughly for later use. 2. For the remaining cell supernatant, take 80μL of each aliquot, add 20μL of 5× SDS Loading Buffer, mix thoroughly, and boil in boiling water for 10 minutes. Centrifuge and load onto the sample or store in a -20°C refrigerator.

[0063] e. Protein concentration determination steps: (1) Preparation of BCA working solution: Based on the number of standards and samples to be tested, calculate the total amount of BCA working solution A and B required. Prepare the working solution at a volume ratio of 50:1 between BCA reagents A and B, vortex and mix thoroughly, and set aside.

[0064] (2) 1× PBS diluted protein standard:

[0065]

[0066] (3) Take 25 μL 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 to create bubbles. Cover the 96-well plate tightly and react in a 37°C incubator for 30 minutes. (4) Take out the 96-well plate and return it to room temperature for 3-5 minutes. Measure the absorbance value of A562 on a microplate 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 in preparation for protein loading.

[0067] 3. SDS-PAGE: (1) Fix the gel plate and prepare 10% SDS-PAGE separation gel.

[0068] Prepare separation gel according to the following table: 10 mL

[0069] 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 4 μL Total 10mL

[0070] (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

[0071] Deionized water 2.77mL 30% (m / v) Acrylamide 830μL 0.5M Tris-HCl (pH 6.8) buffer 1.26mL 10% (m / v) SDS 50 μL 10% (m / v) APS 50 μL TEMED 5μL Total 5mL

[0072] (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.

[0073] (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;

[0074] 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.

[0075] 5. Reagent preparation:

[0076] (1) 10% SDS: Weigh 1 g of high-purity (electrophoresis grade) SDS into a 10 mL centrifuge tube, add approximately 8 mL of deionized water, heat to dissolve, and dilute to 10 mL. Store at room temperature.

[0077] (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.

[0078] (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.

[0079] (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.

[0080] (5) 1.5 mol / L Tris-HCl, 100 mL: Dissolve 18.15 g of Tris in 80 mL of water and adjust the dilution to 8.8 with 4 N HCl. Dose to 100 mL.

[0081] (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.

[0082] (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.

[0083] (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.

[0084] (9) Blocking solution, antibody diluent: Add 5% skim milk powder or BSA to TBST buffer and prepare it before use.

[0085] like Figure 1 The results showed that treatment with 30nM, 100nM and 300nM of RD240404 could effectively downregulate the expression levels of p-STAT3(Y705), p-STAT3(S727) and STAT3's downstream target proteins C-myc and CyclinD1 in MDA-B-468 cells.

[0086] The above results demonstrate that piperazinamide compounds, represented by RD240404, can significantly inhibit the growth of lymphoma, multiple myeloma, leukemia, breast cancer, gastric cancer, lung cancer, and pancreatic cancer cells, and exhibit significant inhibitory effects on STAT3 and its related proteins, demonstrating promising anticancer activity. Following the typical drug development approach (conventional in vitro anti-tumor screening followed by targeted research), the compounds of the present invention can be applied to the development of therapeutics for cancers associated with abnormal cell proliferation. Anti-tumor drugs can be prepared by mixing with human-acceptable salts or pharmaceutical carriers.

[0087] 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 replacements of the present invention and any modifications or partial replacements 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 piperazineamide derivative, characterized in that: The structural formula is shown in general formula I: Wherein, n=1, 2, 3, 4, 5, R1 is selected from 2. The piperazinamide derivative according to claim 1, characterized in that Specifically, the compound has the following structure:

3. A biologically acceptable salt of the piperazinamide 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 piperazinamide 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 and t-BuOK were dissolved in tetrahydrofuran, stirred evenly, and then compound 2 was added. The mixture was stirred and reacted at room temperature until complete. The reaction solution was filtered through diatomaceous earth, washed with ethyl acetate, and the organic phase was subjected to spin column chromatography to obtain compound 3. (2) Compound 3, compound 4 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 compound 5; (3) Compound 5 was dissolved in a mixed solution of ethanol 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 dried by column chromatography to obtain compound 6; (4) Dissolve triphosgene in dichloromethane, add compound 6 and DIEA at -5 to 5°C, react for 0.5 to 1 hour, add compound 7, and stir at room temperature until the reaction is complete; the reaction solution is directly spin-dried and subjected to column chromatography to obtain the compound of formula I.

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

3.

6. Use of the piperazinamide 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, 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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