Carborane-containing isoflavone derivative as well as preparation method and application thereof

Abstract

The invention relates to the technical field of medicinal chemistry and pharmacotherapeutics, and mainly relates to a carborane-containing isoflavone derivative as well as a preparation method and application thereof. The carborane-containing isoflavone derivative provided by the invention is a compound as shown in a formula (I) or pharmaceutically acceptable salt thereof, and also provides a preparation method thereof, the compound shows antitumor activity in related cancer cell lines, overcomes the defect that the active action site and mechanism of the isoflavone compound are not clear, and has a good application prospect. The BNCT nano-material has good targeting property, can realize double functions of BNCT precise positioning treatment and tumor growth and migration inhibition, and has very high clinical application value.

Description

TECHNICAL FIELD

[0001] The present application relates to the technical field of pharmaceutical chemistry and pharmacotherapy, in particular to a carborane-containing isoflavone derivative and a preparation method and application thereof. BACKGROUND

[0002] Flavonoids are an important class of natural products, widely exist in plants such as celery, parsley, red pepper, chamomile, mint and ginkgo leaves, and exist in the form of glycosides and free bodies. Flavonoids, in structure, refer to a series of compounds in which two benzene rings with phenolic hydroxyl groups are connected to each other through a central three-carbon atom. According to the oxidation degree of the central three-carbon chain, the connection position of the B ring, and whether the three-carbon chain forms a ring, the main natural flavonoids can be divided into flavones, isoflavones, flavonols, dihydroflavones, chalcones, aurones, etc.

[0003] At present, many flavonoid drugs are being applied in clinical treatment. For example, Shuxuening tablets, which contain flavones and biflavones, are used for the treatment of coronary heart disease and angina pectoris; Licoxin, a fully synthetic ethoxy flavone, is used in clinic to dilate coronary vessels and increase coronary flow. In addition, some of them have also entered the clinical trial stage, such as quercetin for the treatment of Alzheimer's disease (NCT 04685590); Lariciresin for the treatment of secondary lymphedema of lower extremities (NCT 04360889); Fustin for the treatment of carpal tunnel syndrome (NCT 05416515), etc.

[0004] Isoflavones are an important class of flavonoid natural products. Due to their structural similarity to diethylstilbestrol, they can bind to estrogen receptors and have estrogen-like effects, such as genistein, biochanin A, and daidzein (Whitten PL et al., Environmental Health Perspectives, 2001, 109, 5-20.). In recent years, the activity of isoflavones in anti-tumor research has also attracted much attention. Due to the multi-target characteristics of natural products, they have unique therapeutic potential for tumor, a multi-factor induced systemic disease. According to related pharmacodynamic and clinical studies, many flavonoids are effective for breast cancer, prostate cancer, non-small cell lung cancer, etc., and also show significant synergistic effect in combination with marketed anticancer drugs, enhancing the efficacy.

[0005] Formononetin, a potent fibroblast growth factor 2 (FGFR2) inhibitor, has an IC 50about 4.31 μM, effectively inhibits cell proliferation and tumor growth by blocking the Akt signaling pathway in breast cancer cells; at the same time, it inhibits tumor angiogenesis, and can overcome the drug resistance of traditional vascular endothelial growth factor (VEGF) -targeted angiogenesis inhibitors (Xiaoyu Wu et al., Oncotarget, 2015, 6, 42.). In addition, studies have shown that formononetin can regulate multiple signaling pathways, such as the Janus kinase / signal transducer and activator of transcription (JAK / STAT) signaling pathway, the phosphatidylinositol 3-kinase / protein kinase B (PI3K / AKT) signaling pathway, and the extracellular signal-regulated kinase 1 / 2 (ERK1 / 2) signaling pathway in the mitogen-activated protein kinase (MAPK), inhibiting cell proliferation, invasion, and migration. When combined with other chemotherapeutic drugs such as bortezomib, LY2940002, U0126, sunitinib, epirubicin, doxorubicin, temozolomide, metformin, etc., it can produce a synergistic effect to enhance the anti-cancer effect of formononetin and related drugs (Kai-Ching Tay et al., Frontiers in Pharmacology, 2019, 10, 820.). Psoralen isoflavone is also an important bioactive compound that can inhibit the expression of downstream genes of the transcription factor protein family nuclear factor kappa B (NF-κB) and inhibit the proliferation and migration of non-small cell lung cancer (NSCLC) cells. At the same time, it can significantly reverse the proliferation of NSCLC cell lines induced by p65 overexpression (Zihan Lin et al., Phytomedicine, 2023, 110, 154627.).

[0006] However, based on the characteristics of multi-targets of natural products, there are also some drawbacks, such as the site and mechanism of active action are not clear enough, which limits the further development of natural product drugs. Therefore, it is of great significance to reasonably modify the structure of natural products, improve their targeting, and clarify their distribution in the body. SUMMARY

[0007] In view of the defects of isoflavone compounds in the prior art, the purpose of the present application is to provide a carbon-borane-containing isoflavone derivative and a preparation method and use thereof, which shows anti-tumor activity in related cancer cell lines.

[0008] To achieve this purpose, the technical scheme adopted by the present application is as follows:

[0009] The first aspect of the present application provides a carbon-borane-containing isoflavone derivative, which is a compound represented by formula (I):

[0010]

[0011] wherein R 1 and R2 each independently selected from a hydrogen atom, a hydroxyl group, a benzyloxy group, a methoxy group, a sulfonylfluorooxy group; X is selected from a carbonyl group, a methylene group, CH-OH;

[0012] X-Y is selected from a carbon-carbon single bond or a carbon-carbon double bond; Y-Z is selected from a carbon-carbon single bond or a carbon-carbon double bond.

[0013] The "carborane" in the present application is a polyhedral structure composed of CH and BH vertices, preferably a icosahedral closed-loop dodecaborane structure (C2B 10 H 12 Due to the low polarity of B-H bond, carborane has a high hydrophobic property, which is beneficial to enhance the transport of small molecules across the cell membrane and the blood-brain barrier; at the same time, as an inorganic structure, carborane can avoid being degraded by enzymes in the body, so it has high stability under biological conditions. In addition, the detection background of boron element in the body is extremely low, which provides favorable conditions for analyzing the distribution and pharmacokinetic properties of carborane-modified compounds in the body. The present application connects carborane with natural isoflavone compounds with anti-tumor activity, which can overcome the drawbacks of unclear active site and mechanism of isoflavone compounds, improve the targeting property, and can be used in combination with boron neutron capture therapy (BNCT) to achieve precise positioning treatment and inhibit tumor growth and migration.

[0014] In some embodiments of the present application, the compound represented by formula (I) is selected from the following group:

[0015] The first group: the compound represented by formula D

[0016]

[0017] wherein, R 3 and R 4 each independently selected from a hydrogen atom, a methoxy group, a benzyloxy group;

[0018] The second group: the compound represented by formula E

[0019]

[0020] wherein, R 8 is selected from a hydroxyl group or a methoxy group, and R 9 is selected from a hydrogen atom or a hydroxyl group;

[0021] The third group: the compound represented by formula F

[0022]

[0023] wherein, R 10 is selected from a methoxy group or a hydroxyl group;

[0024] The fourth group: the compound represented by formula G

[0025]

[0026] wherein, R 7 selected from methoxy or sulfonylfluorideoxy; Q is selected from carbonyl, methylene; Q-T is selected from carbon-carbon single bond or carbon-carbon double bond; T-L is selected from carbon-carbon single bond or carbon-carbon double bond.

[0027] In some embodiments of the present application, the compound of formula D is selected from

[0028]

[0029] In some embodiments of the present application, the compound of formula E is selected from

[0030]

[0031] In some embodiments of the present application, the compound of formula F is selected from

[0032]

[0033] In some embodiments of the present application, the compound of formula G is selected from

[0034]

[0035] The second aspect of the present application provides a method for preparing the above-mentioned carbon-borane-containing isoflavone derivatives.

[0036] The method for preparing the above-mentioned first group of carbon-borane-containing isoflavone derivatives comprises the following steps:

[0037] S1, subjecting a compound of formula A to a Sonogashira coupling reaction with trimethylsilyl acetylene in the presence of a palladium catalyst and a cuprous halide to provide a compound of formula B;

[0038] S2, subjecting the compound of formula B to a reaction under acidic conditions by dropwise adding tetrabutylammonium fluoride to remove the TMS protecting group to provide a compound of formula C;

[0039] S3, subjecting the compound C to a reaction with decaboron dodecahydro dicyano complex in the presence of ionic solution (Bmim)Cl to obtain a compound of formula D;

[0040] The reaction scheme is as follows:

[0041]

[0042] wherein, R 3 and R 4 each independently selected from a hydrogen atom, methoxy, benzyloxy, R 5 selected from Br or I.

[0043] In some embodiments of the present application, the palladium catalyst in step S1 is selected from dichlorobis(triphenylphosphine)palladium, the cuprous halide is selected from cuprous iodide; and / or, the compound of formula A is dissolved in an organic solvent before the reaction; and / or, triethylamine is contained in the reaction system; and / or, the molar ratio of the compound of formula A, trimethylsilyl acetylene, palladium catalyst and cuprous halide is 1:2-3:0.04-0.06:0.008-0.012; and / or, the reaction temperature is room temperature or zero degree; and / or, after the reaction is completed, the reaction product is separated and purified, and the method of separation and purification is preferably column chromatography.

[0044] In some embodiments of the present application, the compound of formula B in step S2 is dissolved in an organic solvent before the reaction, and the organic solvent is preferably tetrahydrofuran; and / or, the acidic condition is specifically the addition of d-camphorsulfonic acid; and / or, the molar amount of tetrabutylammonium fluoride added dropwise is twice that of the compound of formula B; and / or, the reaction temperature is room temperature; and / or, after the reaction is completed, water is added to quench, and the reaction product is separated and purified, and the separation and purification includes extraction, washing, drying, concentration, column chromatography.

[0045] In some embodiments of the present application, the compound of formula C in step S3 is dissolved in an organic solvent before the reaction, and the organic solvent is preferably toluene; and / or, the molar ratio of the compound of formula C, ten-boron dodeca-hydrodiacetonitrile complex is 3:3-6; and / or, the reaction temperature is 105-115°C, and the reaction time is 2-6h; and / or, after the reaction is completed, the reaction product is separated and purified, and the method of separation and purification is preferably column chromatography.

[0046] The preparation method of the second group of carbon-borane-containing isoflavone derivatives described above comprises the following steps: R 3 is a methoxy group, R 4 The compound of formula D selected from a hydrogen atom or a methoxy group is added dropwise with a solution of boron tribromide under ice bath conditions, and after the reaction is completed, saturated aqueous ammonium chloride solution is added to quench, to provide a compound of formula E;

[0047] The reaction formula is as follows:

[0048]

[0049] wherein, R 3 is a methoxy group, R 4 is selected from a hydrogen atom or a methoxy group, R 8 is selected from a hydroxyl group or a methoxy group, R 9 is selected from a hydrogen atom or a hydroxyl group.

[0050] In some embodiments of the present application, R 3 is a methoxy group, R 4The compound of formula D selected from a hydrogen atom or methoxy is dissolved in an organic solvent, preferably 1,2-dichloroethane, and then participates in the reaction; and / or, the molar ratio of the compound of formula D selected from a hydrogen atom or methoxy to boron tribromide is 1:1-4; and / or, the reaction temperature is 65-75°C, and the reaction time is 10-14h; and / or, the reaction product is separated and purified, which includes extraction, washing, drying, concentration, and column chromatography. 3 R is a methoxy group, R 4 The molar ratio of the compound of formula D selected from a hydrogen atom or methoxy to boron tribromide is 1:1-4; and / or, the reaction temperature is 65-75°C, and the reaction time is 10-14h; and / or, the reaction product is separated and purified, which includes extraction, washing, drying, concentration, and column chromatography.

[0051] The preparation method of the third group of carbon-borane-containing isoflavone derivatives described above comprises the following steps: R 8 R is selected from a methoxy group or a hydroxyl group, R 9 The compound of formula E in which R

[0052] The reaction formula is as follows:

[0053]

[0054] R is selected from a methoxy group or a hydroxyl group, R 8 R is selected from a methoxy group or a hydroxyl group, R 9 R is a hydrogen atom, R 10 R is selected from a methoxy group or a hydroxyl group.

[0055] In some embodiments of the present application, R 8 R is selected from a methoxy group or a hydroxyl group, R 9 The compound of formula E in which R is a hydrogen atom is dissolved in an organic solvent, preferably tetrahydrofuran, and then participates in the reaction; and / or, the borohydride reagent is sodium borohydride, and the sodium borohydride is added under ice bath conditions, R 8 R is selected from a methoxy group or a hydroxyl group, R 9 The molar ratio of the compound of formula E in which R is a hydrogen atom to sodium borohydride is 1:1-4; and / or, the reaction temperature is room temperature, and the reaction time is 6-12h; and / or, after the reaction is completed, water is added for quenching; and / or, the reaction product is separated and purified, which includes extraction, washing, drying, concentration, and column chromatography.

[0056] The preparation method of the fourth group of carbon-borane-containing isoflavone derivatives described above comprises any one of the following steps:

[0057] (a) The compound of formula F in which R 10 The compound of formula F in which R is a methoxy group is subjected to an oxidation reaction in the presence of manganese dioxide to provide R 7 The compound of formula G in which R is a methoxy group, Q is a carbonyl group, Q-T is a carbon-carbon single bond, and T-L is a carbon-carbon single bond;

[0058] The reaction formula is as follows:

[0059]

[0060] In some embodiments of the present invention, the R mentioned in step (a) above 10 The methoxylated compound of formula F participates in the reaction after being dissolved in an organic solvent, preferably dichloromethane; and / or, the R... 10 The molar ratio of the methoxy compound of formula F to manganese dioxide is 1:1 to 4; and / or the reaction temperature is room temperature and the reaction time is 6 to 12 h; and / or, after the reaction is completed, the reaction product is separated and purified, the separation and purification including filtration, washing, concentration and column chromatography.

[0061] (b) R 10 The methoxylated compound of formula F reacts with polyphosphoric acid, and after the reaction is complete, it is quenched with water and purified to provide R. 7 Compound G has the following structure: Q is methoxy, Q is methylene, QT is carbon-carbon double bond, and TL is carbon-carbon single bond.

[0062] The reaction formula is as follows:

[0063]

[0064] In some embodiments of the present invention, R in step (b) above 10 The methoxylated compound of formula F participates in the reaction after being dissolved in an organic solvent, preferably dichloromethane; and / or, the R... 10 The ratio of the methoxylated compound of formula F to polyphosphoric acid is 1 mmol: 300–800 mg; and / or, the reaction temperature is 75–85 °C and the reaction time is 6–12 h; and / or, the reaction product is separated and purified, the separation and purification including extraction, washing, drying, concentration, and column chromatography.

[0065] (c) R 10 The methoxylated compound of formula F reacts with polyphosphoric acid, and after the reaction is complete, it is quenched with water and purified to provide R. 7 Compound G is a compound with a methoxy group, Q is a methylene group, QT is a carbon-carbon double bond, and TL is a carbon-carbon single bond; the resulting R... 7 Compound G, consisting of a methoxy group, Q being a methylene group, QT being a carbon-carbon double bond, and TL being a carbon-carbon single bond, undergoes an addition reaction with hydrogen in the presence of a palladium-on-carbon catalyst to provide R. 7 The compound of formula G has methoxy group, Q is methylene group, QT is carbon-carbon single bond, and TL is carbon-carbon single bond.

[0066] The reaction formula is as follows:

[0067]

[0068] In some embodiments of the present invention, the R mentioned in step (c) above 10 The methoxylated compound of formula F is dissolved in an organic solvent, preferably dichloromethane, to participate in the first step reaction; and / or, the R... 10 The ratio of methoxylated compound F to polyphosphoric acid is 1 mmol: 300–800 mg; and / or, the temperature of the first step reaction is 75–85 °C, and the reaction time is 6–12 h; and / or, the product of the first step reaction is separated and purified, the separation and purification including extraction, washing, drying, concentration, and column chromatography; and / or, the R in step (c) above... 7 The compound of formula G, where Q is methoxy, Q is methylene, QT is a carbon-carbon double bond, and TL is a carbon-carbon single bond, is dissolved in an organic solvent and then participates in the second step reaction. The organic solvent is preferably tetrahydrofuran. And / or, the palladium on carbon is 5% palladium on carbon (55% water), and the ratio of the compound of formula L to palladium on carbon is 1 mmol: 50–200 mg. And / or, the temperature of the second step reaction is room temperature, and the reaction time is 5–7 h. And / or, the product of the second step reaction is separated and purified, preferably by column chromatography.

[0069] (d) The above R will be adopted 8 For hydroxyl group, R 9 Compound E, with hydrogen atoms, reacts with 1-(fluorosulfonyl)-2,3-dimethyl-1H-imidazolium-3-onium trifluoromethanesulfonate in the presence of a base to provide R. 7 It is a compound of formula G, where Q is a sulfonyl fluoride group, Q is a carbonyl group, QT is a carbon-carbon single bond, and TL is a carbon-carbon double bond;

[0070] The reaction formula is as follows:

[0071]

[0072] In some embodiments of the present invention, the R mentioned in step (d) above... 8 For hydroxyl group, R 9 Compound E, which contains hydrogen atoms, participates in the reaction after being dissolved in an organic solvent, preferably acetonitrile; and / or, R... 8 For hydroxyl group, R 9 The molar ratio of the compound of formula E (containing hydrogen atoms) and 1-(fluorosulfonyl)-2,3-dimethyl-1H-imidazolium-3-onium trifluoromethanesulfonate is 1:1 to 5; and / or, the base is selected from triethylamine; and / or, the reaction temperature is room temperature and the reaction time is 0.8 to 1.2 h; and / or, the reaction product is separated and purified, preferably by column chromatography.

[0073] A third aspect of this invention provides the use of the above-mentioned carborane-containing isoflavone derivatives or pharmaceutically acceptable salts thereof in the preparation of antitumor drugs. The tumors include, but are not limited to, breast cancer, prostate cancer, lung cancer, squamous cell carcinoma, pancreatic cancer, gastric cancer, colon cancer, diffuse large B-cell lymphoma, chronic myeloid leukemia, and acute myeloid leukemia. In particular, the antitumor drug is an anti-multidrug-resistant tumor drug. The multidrug-resistant tumors include, but are not limited to, multidrug-resistant chronic leukemia.

[0074] As can be seen from the cell experiment results of the embodiments of the present invention, the carborane-containing isoflavone derivatives provided by the present invention have proliferative inhibitory activity against a variety of tumor cells and multidrug-resistant tumor cells.

[0075] Compared with the prior art, the present invention has the following advantages:

[0076] 1. The carborane-containing isoflavone derivatives provided by this invention link isoflavone natural products with antitumor activity with carborane, a novel boron carrier, overcoming the drawbacks of unclear active sites and mechanisms of isoflavone compounds. It has good targeting and can achieve the dual effects of precise BNCT treatment and inhibition of tumor growth and migration, and has high clinical application value.

[0077] 2. The method for preparing carborane isoflavone derivatives provided by the present invention has the advantages of readily available raw materials, simple operation, low cost, and environmental friendliness. Each raw material can be obtained cheaply through commercial channels or synthesized through simple steps, and has good prospects for industrialization.

[0078] 3. The carborane-containing isoflavone derivatives provided by this invention can inhibit the proliferation of tumor cells and have excellent anti-tumor and anti-multidrug resistance effects. Attached Figure Description

[0079] Figure 1 The effect of the compounds in this embodiment on the expression of MDR1 in multidrug-resistant K562 / ADR cells is shown in the figure, where A is ZYR-FA-4 and B is ZYR-FA-13. Detailed Implementation

[0080] The following details the carborane-containing isoflavone derivatives of the present invention, their preparation methods, and their uses.

[0081] This invention relates to the preparation of carborane-containing isoflavone derivatives by linking a novel boron-supported carborane with isoflavone natural products possessing antitumor activity. The antitumor activity of these derivatives was studied, and the results showed that the carborane-containing isoflavone derivatives exhibited proliferative inhibitory activity against various tumor cell lines and also showed significant inhibitory effects against multidrug-resistant tumor cells. Based on this, the invention was completed.

[0082] The following detailed description of specific embodiments of the present invention, in conjunction with preferred embodiments, further illustrates the relevant details. When numerical ranges are given in the embodiments, it should be understood that, unless otherwise specified in the present invention, both endpoints of each numerical range, as well as any value between the two endpoints, may be selected. Unless otherwise defined, all technical and scientific terms used in this invention have the same meaning as commonly understood by those skilled in the art. In addition to the specific methods, devices, and materials used in the embodiments, the present invention can be implemented using any prior art methods, devices, and materials similar to or equivalent to those described in the embodiments of the present invention, provided that those skilled in the art possess the prior art and the description of the present invention.

[0083] Unless otherwise stated, the experimental methods, detection methods, and preparation methods disclosed in this invention all employ conventional techniques in the fields of molecular biology, biochemistry, chromatin structure and analysis, analytical chemistry, cell culture, and related areas. Unless otherwise stated, all materials and equipment used in this invention are commercially available.

[0084] Example 1: Preparation of compound 1-4 (Z-FA-1)

[0085]

[0086] Step 1: Dissolve 3-bromotryptone 1-1 (562.6 mg, 2.5 mmol) in 8 mL of triethylamine, add trimethylsilylacetylene (745 μL, 5 mmol), bis(triphenylphosphine)palladium dichloride (87.7 mg, 0.125 mmol), and cuprous iodide (4.76 mg, 0.025 mmol), and stir overnight at room temperature. After the reaction is complete as monitored by TLC, obtain a brown solid 1-2 (272.9 mg, 45% yield) by silica gel column chromatography (petroleum ether: ethyl acetate = 20:1). 1 HNMR (500MHz, CDCl3) δ8.25 (dd, J=8.0, 1.0Hz, 1H), 8.12 (s, 1H), 7.70-7.66 (m, 1H), 7.47-7.42 (m, 2H), 0.27 (s, 9H).

[0087] Step 2: Dissolve 1-2 (272.9 mg, 1.1 mmol) in 3 mL of tetrahydrofuran, add dextrorotatory camphor sulfonic acid (293.2 mg, 1.3 mmol), and dropwise add tetrabutylammonium fluoride (1.0 M in THF, 2.2 mL, 2.2 mmol). Stir the reaction at room temperature for 30 minutes. After the reaction is complete as monitored by TLC, quench with water, extract three times with ethyl acetate (3 × 20 mL), wash with saturated sodium chloride solution, dry to anhydrous sodium sulfate, and concentrate. Analyze by silica gel column chromatography (petroleum ether:ethyl acetate = 20:1) to give brown solid 1-3 (151.2 mg, yield 65%). 1 H NMR (500MHz, CDCl3) δ8.27 (dd, J=8.0, 1.0Hz, 1H), 8.20 (s, 1H), 7.72-7.68 (m, 1H), 7.50-7.41 (m, 2H), 3.28 (s, 1H).

[0088] Step 3: Dissolve 1-3 (151.2 mg, 0.9 mmol) in 3 mL of toluene, add decaboron-dodecyl diacetonitrile complex (244.8 mg, 1.2 mmol) and ionic solution (Bmim) Cl (34 mg, 0.2 mmol). Stir at 110 °C for 3 hours. After the reaction is complete as monitored by TLC, the solution is purified by silica gel column chromatography (petroleum ether: ethyl acetate = 30:1) to give an off-white solid 1-4 (72 mg, 30% yield). 1 H NMR (500MHz, CDCl3) δ8.28 (s, 1H), 8.18 (d, J = 8.0Hz, 1H), 7.75-7.72 (m, 1H), 7.50-7.46 (m, 2H), 6.18 (s, 1H), 3.02-1.71 (br, 10H).

[0089] Example 2: Preparation of compounds 2-6 (Z-FA-2) and 2-7 (Z-FA-4)

[0090]

[0091] Step 1: Dissolve 2-1 (830.8 mg, 5.0 mmol) in 15 mL of N,N-dimethylformamide, add N,N-dimethylformamide dimethyl acetal (1.3 mL, 10.0 mmol), and react at 70 °C for 4 hours. After the reaction is completed by TLC monitoring, quench with water, extract with ethyl acetate (3 × 100 mL), wash with saturated sodium chloride solution, dry to anhydrous sodium sulfate, and concentrate. Analyze by silica gel column chromatography (petroleum ether:ethyl acetate = 8:1) to give a yellow solid 2-2 (1.1 g, 99% yield). 1H NMR(500MHz, CDCl3) δ7.84(d,J=11.5Hz,1H),7.60(d,J=9.0Hz,1H),6.42(d,J=2.5Hz,1H), 6.38(dd,J=9.0,2.5Hz,1H),5.68(d,J=11.5Hz,1H),3.81(s,3H),3.17(s,3H),2.96(s,3H).

[0092] Step 2: Dissolve 2-2 (1.1 g, 5.0 mmol) in 20 mL of methanol, add elemental iodine (2.5 g, 10.0 mmol), and heat to 40 °C for 2 hours. After the reaction is complete as monitored by TLC, quench with saturated sodium thiosulfate solution, extract with ethyl acetate (3 × 100 mL), wash with saturated sodium chloride solution, dry to anhydrous sodium sulfate, and concentrate. Analyze by silica gel column chromatography (petroleum ether:ethyl acetate = 5:1) to give a pale yellow solid 2-3 (1.3 g, yield 83%). 1 H NMR (500MHz, CDCl3) δ8.21(s,1H),8.15(d,J=9.0Hz,1H),7.01(d,J=9.0,2.5Hz,1H),6.84(d,J=2.5Hz,1H),3.91(s,3H).

[0093] Step 3: Dissolve 2-3 (1.3 g, 4.2 mmol) in 10 mL of tetrahydrofuran solution. Under ice bath conditions, add bis(triphenylphosphine)palladium dichloride (147.4 mg, 0.21 mmol), cuprous iodide (80 mg, 0.42 mmol), trimethylsilylacetylene (760 μL, 5.1 mmol), and triethylamine (896 μL, 6.72 mmol) sequentially. Transfer the reaction mixture to room temperature and stir for 3 hours. After the reaction is complete as monitored by TLC, obtain a brown solid 2-4 (924 mg, 83% yield) by silica gel column chromatography (petroleum ether:ethyl acetate = 5:1). 1 HNMR (500MHz, CDCl3) δ8.14(d,J=9.0Hz,1H),8.10(s,1H),6.98(dd,J=9.0,2.5Hz,1H),6.83(d,J=2.5Hz,1H),3.90(s,3H),0.26(s,9H).

[0094] Step 4: Dissolve 2-4 (924 mg, 3.4 mmol) in 10 mL of tetrahydrofuran, add dextrorotatory camphor sulfonic acid (946 mg, 4.1 mmol), and add tetrabutylammonium fluoride (1.0 M in THF, 6.8 mL, 6.8 mmol). Stir the reaction at room temperature for 30 minutes. After the reaction is complete as monitored by TLC, quench with water, extract three times with ethyl acetate (3 × 20 mL), wash with saturated sodium chloride solution, dry to anhydrous sodium sulfate, and concentrate. Analyze by silica gel column chromatography (petroleum ether:ethyl acetate = 5:1) to give a brown solid 2-5 (646 mg, 95% yield). 1 H NMR (500MHz, CDCl3) δ8.16(d,J=9.0Hz,1H),8.12(s,1H),6.99(dd,J=9.0,2.5Hz,1H),6.84(d,J=2.5Hz,1H),3.91(s,3H),3.26(s,1H).

[0095] Step 5: Dissolve 2-5 (646 mg, 3.23 mmol) in 10 mL of toluene, add decaboron-dodecyl diacetonitrile complex (857 mg, 4.2 mmol) and ionic solution (Bmim) Cl (110 mg, 0.65 mmol), and heat to 110 °C under reflux for 3 hours. After the reaction is completed by TLC monitoring, the white solid 2-6 (406 mg, yield 40%) is obtained by silica gel column chromatography (petroleum ether: ethyl acetate = 10:1). 1 H NMR (500MHz, CDCl3) δ8.18(s,1H),8.06(d,J=9.0Hz,1H),7.02(dd,J=9.0,2.5 Hz,1H),6.84(d,J=2.5Hz,1H),6.24(s,1H),3.92(s,3H),3.10-1.82(br,10H).

[0096] Step 6: Dissolve 2-6 (406 mg, 1.3 mmol) in 5 mL of 1,2-dichloroethane, add boron tribromide solution (2.0 M in DCM, 1.3 mL, 2.6 mmol) dropwise under ice bath conditions, and then heat to 70 °C and reflux overnight. After the reaction is completed by TLC monitoring, quench with saturated ammonium chloride aqueous solution, extract three times with ethyl acetate (3 × 10 mL), wash with saturated sodium chloride solution, dry to anhydrous sodium sulfate, concentrate, and obtain a white solid 2-7 (350 mg, 89% yield) by silica gel column chromatography (petroleum ether:ethyl acetate = 5:1). 1HNMR(500MHz, CDCl3)δ8.20(s,1H),8.05(d,J=8.5Hz,1H),6.95(d,J=8.5Hz,1 H),6.86(s,1H),6.29(s,1H),6.18(s,1H),3.85(s,3H),3.06-1.61(br,10H).

[0097] Example 3: Preparation of compounds 2-8 (Z-FA-8), 2-9 (Z-FA-9), 2-10 (Z-FA-10) and 2-11 (Z-FA-11)

[0098]

[0099] Step 1: Dissolve 2-6 (96 mg, 0.3 mmol) in 1 mL of tetrahydrofuran, add sodium borohydride (23 mg, 0.6 mmol) under ice bath conditions, and react at room temperature for 8 hours. After the reaction is completed by TLC monitoring, quench with water, extract three times with ethyl acetate (3 × 5 mL), wash with saturated sodium chloride solution, dry to anhydrous sodium sulfate, concentrate, and obtain a white solid 2-8 (65 mg, yield 67%) by silica gel column chromatography (petroleum ether: ethyl acetate = 5:1). 1 HNMR (500MHz, CDCl3) δ7.08(d,J=8.5Hz,1H),6.52(dd,J=2.5,8.5Hz,1H),6.38(d,J=2.5Hz,1H),4.87(s,1H),4.59(s ,1H),4.37(dd,J=2.0,11.0Hz,1H),4.19(t,J=2.0Hz,1H),3.77(s,3H),2.83(d,J=11.0Hz,1H),2.80-1.76(br,10H).

[0100] Step 2: Dissolve 2-8 (65 mg, 0.2 mmol) in 1 mL of dichloromethane, add manganese dioxide (44 mg, 0.5 mmol), and react at room temperature for 8 hours. After the reaction is complete as monitored by TLC, filter to remove the solid, wash with dichloromethane, and concentrate. Analyze by silica gel column chromatography (petroleum ether: ethyl acetate = 5:1) to give a white solid 2-9 (60 mg, 98% yield). 1H NMR (500MHz, CDCl3) δ7.85(d,J=8.5Hz,1H),6.64(dd,J=2.0,8.5Hz,1H),6.42(d,J=2Hz,1H),4.65(dd,J =4.5,12.0Hz,1H),4.56(s,J=4.5,12.0Hz,1H),4.46(s,1H),3.29(t,J=4.5Hz,1H),2.97-1.66(br,10H).

[0101] Step 3: Dissolve 2-8 (65 mg, 0.2 mmol) in 1 mL of dichloroethane, add polyphosphoric acid (100 mg), and reflux at 80 °C for 8 hours. After the reaction is completed by TLC monitoring, quench with water, extract three times with dichloromethane (3 × 5 mL), wash with saturated sodium chloride solution, dry to anhydrous sodium sulfate, concentrate, and obtain a white solid 2-10 (37 mg, yield 56%) by silica gel column chromatography (petroleum ether: ethyl acetate = 8:1). 1 HNMR(500MHz, CDCl3)δ6.94(d,J=8.5Hz,1H),6.63(s,1H),6.47(dd,J=2.5,8.5Hz,1H ),6.38(d,J=2.5Hz,1H),4.73(s,2H),3.81(s,1H),3.78(s,3H),2.84-1.76(br,10H).

[0102] Step 4: Dissolve 2-10 (37 mg, 0.1 mmol) in 1 mL of tetrahydrofuran, add 5% palladium on carbon (55% water, 10 mg), then evacuate the reaction system, purge with hydrogen, and react at room temperature for 6 hours. After the reaction is complete as monitored by TLC, obtain a white solid 2-11 (35 mg, 98% yield) by silica gel column chromatography (petroleum ether: ethyl acetate = 8:1). 1 HNMR (500MHz, CDCl3) δ6.93 (d, J=8.5Hz, 1H), 6.49 (dd, J=2.5, 8.5Hz, 1H), 6.36 (d, J=2.5Hz, 1H),4.35(m,1H),3.81(m,1H),3.75(m,4H),2.90(m,1H),2.77(m,2H),2.75-1.72(br,10H).

[0103] Example 4: Preparation of compounds 2-12 (Z-FA-12) and 2-13 (Z-FA-13)

[0104]

[0105] Step 1: Dissolve 2-7 (350 mg, 1.2 mmol) in 5 mL of tetrahydrofuran, add sodium borohydride (92 mg, 2.4 mmol) under ice bath conditions, and react at room temperature for 8 hours. After the reaction is completed by TLC monitoring, the white solid 2-12 (189 mg, yield 45%) is obtained by silica gel column chromatography (petroleum ether: ethyl acetate = 5:1). 1 H NMR (500MHz, CDCl3) δ7.04(d,J=8.5Hz,1H),6.44(dd,J=2.5,8.5Hz,1H),6.32(d,J=2.5Hz,1H),5.33(s,1H ),4.85(s,1H),4.58(s,1H),4.35-4.35(m,1H),4.20-4.16(m,1H),2.83-2.80(m,1H),2.80-1.79(br,10H).

[0106] Step 2: Dissolve 2-7 (350 mg, 1.2 mmol) in 5 mL of acetonitrile, add 1-(fluorosulfonyl)-2,3-dimethyl-1H-imidazolium-3-onium trifluoromethanesulfonate (945 mg, 3.0 mmol) and triethylamine (240 μL, 1.8 mmol), and react at room temperature for 1 hour. After the reaction is completed by TLC monitoring, the white solid 2-13 (362 mg, 78% yield) is obtained by silica gel column chromatography (petroleum ether: ethyl acetate = 15:1). 1 HNMR (500MHz, CDCl3) δ8.33(d,J=9.0Hz,1H),8.31(s,1H),6.54(d,J=2.0Hz,1H),7.46(dd,J=2.0,9.0Hz,1H),6.06(s,1H),3.02-1.71(br,10H).

[0107] Example 5: Preparation of compounds 3-6 (Z-FA-5), 3-7 (Z-FA-6), and 3-8 (Z-FA-7)

[0108]

[0109] Step 1: Dissolve 3-1 (980 mg, 5.0 mmol) in 15 mL of N,N-dimethylformamide, then add N,N-dimethylformamide dimethyl acetal (1.3 mL, 10.0 mmol), and react at 70 °C for 4 hours. After the reaction is completed by TLC monitoring, quench with water, extract with ethyl acetate (3 × 100 mL), wash with saturated sodium chloride solution, dry to anhydrous sodium sulfate, and concentrate. Analyze by silica gel column chromatography (petroleum ether:ethyl acetate = 2:1) to give a yellow solid 3-2 (1.2 g, 99% yield). 1HNMR (500MHz, CDCl3) δ15.62(s,1H),7.90(d,J=12.0Hz,1H),6.24(d,J=12.0Hz,1H),6.06( d,J=2.0Hz,1H),5.90(d,J=2.0Hz,1H),3.83(s,3H),3.78(s,3H),3.13(s,3H),2.92(s,3H).

[0110] Step 2: Dissolve 3-2 (1.2 g, 5.0 mmol) in 20 mL of methanol, add elemental iodine (2.5 g, 10.0 mmol), and react at room temperature for 24 hours. After the reaction is completed by TLC monitoring, quench with water, extract with ethyl acetate (3 × 100 mL), wash with saturated sodium chloride solution, dry to anhydrous sodium sulfate, and concentrate. Analyze by silica gel column chromatography (petroleum ether:ethyl acetate = 2:1) to give a pale yellow solid 3-3 (697 mg, yield 42%). 1 HNMR (500MHz, CDCl3) δ8.08 (s, 1H), 6.43 (d, J = 2.0Hz, 1H), 6.39 (d, J = 2.0Hz, 1H), 3.93 (s, 3H), 3.88 (s, 3H).

[0111] Step 3: Dissolve 3-3 (697 mg, 2.1 mmol) in 10 mL of tetrahydrofuran solution. Under ice bath conditions, add bis(triphenylphosphine)palladium dichloride (148 mg, 0.2 mmol), cuprous iodide (80 mg, 0.4 mmol), trimethylsilylacetylene (376 μL, 2.6 mmol), and triethylamine (448 μL, 3.4 mmol) sequentially. Transfer the reaction mixture to room temperature and stir for 3 hours. After the reaction is complete as monitored by TLC, obtain a brown solid 3-4 (272 mg, 45% yield) by silica gel column chromatography (petroleum ether:ethyl acetate = 2:1). 1 H NMR (500MHz, CDCl3) δ7.94(s,1H),6.40(s,1H),6.35(s,1H),3.91(s,3H),3.87(s,3H),0.24(s,9H).

[0112] Step 4: Dissolve 3-4 (286 mg, 0.9 mmol) in 3 mL of tetrahydrofuran, add dextrorotatory camphor sulfonic acid (209 mg, 1.1 mmol), and dropwise add tetrabutylammonium fluoride (1.0 M in THF, 1.8 mL, 1.8 mmol). Stir the reaction at room temperature for 30 minutes. After the reaction is complete as monitored by TLC, quench with water, extract with ethyl acetate (3 × 10 mL), wash with saturated sodium chloride solution, dry to anhydrous sodium sulfate, and concentrate. Analyze by silica gel column chromatography (petroleum ether:ethyl acetate = 5:1) to give a brown solid 3-5 (196 mg, 95% yield).1 HNMR (500MHz, CDCl3) δ7.97(s,1H),6.42(d,J=2.0Hz,1H),6.37(d,J=2.0Hz,1H),3.93(s,3H),3.88(s,3H),3.21(s,1H).

[0113] Step 5: Dissolve 3-5 (196 mg, 0.9 mmol) in 5 mL of toluene, add decaboron-dodecyl diacetonitrile complex (247 mg, 1.2 mmol) and ionic solution (Bmim) Cl (32 mg, 0.19 mmol), and heat to 110 °C under reflux for 3 hours. After the reaction is completed by TLC monitoring, the white solid 3-6 (152 mg, yield 48%) is obtained by silica gel column chromatography (petroleum ether: ethyl acetate = 5:1). 1 H NMR (500MHz, CDCl3) δ8.06(s,1H),6.44(s,1H),6.40(s,1H),6.30(s,1H),3.94(s,3H),3.89(s,3H),3.01-1.72(br,10H).

[0114] Step 6: Dissolve 3-6 (152 mg, 0.4 mmol) in 2 mL of dichloromethane, add boron tribromide solution (2.0 M in DCM, 0.4 mL, 0.8 mmol) dropwise under ice bath conditions, then heat to 40 °C and reflux overnight. After the reaction is completed by TLC monitoring, quench with saturated ammonium chloride aqueous solution, extract three times with ethyl acetate (3 × 10 mL), wash with saturated sodium chloride solution, dry to anhydrous sodium sulfate, concentrate, and obtain a white solid 3-7 (119 mg, 89% yield) by silica gel column chromatography (petroleum ether: ethyl acetate = 8:1). 1 H NMR (500MHz, CDCl3) δ12.10(s,1H),8.12(s,1H),6.40(m,2H),6.02(s,1H),3.87(s,1H),3.15-1.76(br,10H).

[0115] Step 7: Dissolve 3-6 (152 mg, 0.4 mmol) in 2 mL of 1,2-dichloroethane, add boron tribromide solution (2.0 M in DCM, 0.8 mL, 1.6 mmol) dropwise under ice bath conditions, and then heat to 70 °C and reflux overnight. After the reaction is completed by TLC monitoring, quench with saturated ammonium chloride aqueous solution, extract three times with ethyl acetate (3 × 10 mL), wash with saturated sodium chloride solution, dry to anhydrous sodium sulfate, concentrate, and obtain a white solid 3-8 (119 mg, 75% yield) by silica gel column chromatography (petroleum ether: ethyl acetate = 5:1). 1HNMR (500MHz, CDCl3) δ12.16(s,1H),8.11(s,1H),6.40(d,J=2.0Hz,1H),6.32(d,J=2.0Hz,1H),6.27(s,1H),6.01(s,1H),3.08-1.62(br,10H).

[0116] Example 6: Preparation of compound 4-6 (Z-FA-3)

[0117]

[0118] Step 1: Dissolve 4-1 (1.2 g, 5.0 mmol) in 15 mL of N,N-dimethylformamide, then add N,N-dimethylformamide dimethyl acetal (1.3 mL, 10.0 mmol), and react at 70 °C for 4 hours. After the reaction is completed by TLC monitoring, quench with water, extract with ethyl acetate (3 × 100 mL), wash with saturated sodium chloride solution, dry to anhydrous sodium sulfate, and concentrate. Analyze by silica gel column chromatography (petroleum ether:ethyl acetate = 5:1) to give a yellow solid 4-2 (1.4 g, 99% yield). 1 H NMR (500MHz, CDCl3) δ7.84(d,J=10.0Hz,1H),7.61(d,J=8.5Hz,1H),7.43-7.31(m,5H),6.50(d,J=2 .0Hz,1H),6.45(dd,J=2.0,8.5Hz,1H),5.68(J=10.0Hz,1H),5.07(s,2H),3.16(s,3H),2.96(s,3H).

[0119] Step 2: Dissolve 3-2 (1.4 g, 5.0 mmol) in 20 mL of methanol, add elemental iodine (2.5 g, 10.0 mmol), and react at room temperature for 24 hours. After the reaction is completed by TLC monitoring, quench with water, extract with ethyl acetate (3 × 100 mL), wash with saturated sodium chloride solution, dry to anhydrous sodium sulfate, and concentrate. Analyze by silica gel column chromatography (petroleum ether:ethyl acetate = 4:1) to give a pale yellow solid 4-3 (1.6 g, yield 89%). 1 H NMR (500MHz, CDCl3) δ8.20 (s, 1H), 8.16 (d, J = 9.0Hz, 1H), 7.45-7.35 (m, 5H), 7.08 (dd, J = 2.0, 9.0Hz, 1H), 6.91 (d, J = 2.0Hz, 1H), 5.16 (s, 2H).

[0120] Step 3: Dissolve 4-3 (1.6 g, 4.4 mmol) in 10 mL of tetrahydrofuran solution. Under ice bath conditions, add bis(triphenylphosphine)palladium dichloride (351 mg, 0.5 mmol), cuprous iodide (95 mg, 0.5 mmol), trimethylsilylacetylene (787 μL, 5.3 mmol), and triethylamine (947 μL, 7.1 mmol) sequentially. Transfer the reaction mixture to room temperature and stir for 3 hours. After the reaction is complete as monitored by TLC, obtain a brown solid 4-4 (1.1 g, 73% yield) by silica gel column chromatography (petroleum ether:ethyl acetate = 4:1). 1 H NMR (500MHz, CDCl3) δ8.15(d,J=9.0Hz,1H),8.09(s,1H),7.42-7.37(m,5H),7.06(d,J=9.0Hz,1H),6.90(s,1H),5.15(s,2H),0.26(s,9H).

[0121] Step 4: Dissolve 4-4 (1.1 g, 3.2 mmol) in 10 mL of tetrahydrofuran, add dextrorotatory camphor sulfonic acid (890 mg, 3.8 mmol), and dropwise add tetrabutylammonium fluoride (1.0 M in THF, 6.5 mL, 6.5 mmol). Stir the reaction at room temperature for 30 minutes. After the reaction is complete as monitored by TLC, quench with water, extract with ethyl acetate (3 × 20 mL), wash with saturated sodium chloride solution, dry to anhydrous sodium sulfate, and concentrate. Analyze by silica gel column chromatography (petroleum ether:ethyl acetate = 3:1) to give a brown solid 4-5 (798 mg, 93% yield). 1 HNMR(500MHz, CDCl3)δ8.17(d,J=9.0Hz,1H),8.11(s,1H),7.45-7.37(m,5H) ,7.07(d,J=2.0,9.0Hz,1H),6.91(d,J=2.0Hz,1H),5.16(s,2H),3.26(s,1H).

[0122] Step 5: Dissolve 4-5 (798 mg, 2.9 mmol) in 10 mL of toluene, add the decaboron-dodecyl diacetonitrile complex (710 mg, 3.5 mmol) and ionic solution (Bmim) Cl (50 mg, 0.29 mmol), and heat to 110 °C under reflux for 3 hours. After the reaction is completed by TLC monitoring, the white solid 4-6 (515 mg, yield 45%) is obtained by silica gel column chromatography (petroleum ether: ethyl acetate = 5:1). 1H NMR(500MHz, CDCl3)δ8.17(d,J=9.0Hz,1H),8.07(s,1H),7.43-7.42(m,5H),7.09(d, J=9.0Hz,1H),6.91(d,J=2.5Hz,1H),6.24(s,1H),5.17(s,2H),2.92-1.69(br,10H).

[0123] Example 7: Inhibitory activity of the compounds of the present invention against tumor cell proliferation

[0124] The inhibitory activity of the compounds on tumor cell proliferation was determined using the human breast cancer cell line MDA-MB-231 and the human prostate cancer cell line PC-3. Cells were grown in DMEM medium containing 10% fetal bovine serum and 1% penicillin-dextrose antibody at 37°C and 5% CO2.

[0125] Experimental methods

[0126] Take cells in good logarithmic growth phase and use 8 × 10⁸ cells. 3 Cells were seeded at a density of [number] cells / well in 96-well plates and cultured for 16 hours using standard methods. Then, different concentrations of the compound of this invention were added sequentially: 100 μM, 30 μM, 10 μM, 3 μM, 1 μM, 300 nM, 100 nM, 30 nM, and 10 nM. An equal volume of DMSO was added to the control group. After culturing for another 120 hours, 100 μL of pre-cooled trichloroacetic acid (50%, w / v) was added to each well, and the cells were incubated at 4°C for 1 hour to fix them. After fixation, the cells were washed five times with distilled water and air-dried. 100 μL of SRB staining solution (4%, w / v) was added to each well, and the cells were incubated at room temperature for 30 minutes to stain. The staining solution was removed, and the cells were washed five times with 1% acetic acid to remove unbound dye. After air-drying, 150 μL of 10 mM Tris solution was added to each well, and the bound SRB dye was dissolved by shaking. Place the 96-well plate in a microplate reader and measure the OD value at a wavelength of 530 nm. Calculate the cell viability (%) according to the formula below.

[0127]

[0128] Statistical analysis was performed on the effects of different concentrations of the compounds of this invention on the proliferation of breast cancer and prostate cancer cells. GraphPadPrism software was used for IC50 analysis of different compounds. 50 The calculations and test results are shown in Table 1.

[0129] Experimental results

[0130] The results, as shown in Table 1, indicate that the compounds of this invention exhibit significant inhibitory activity against the proliferation of human breast cancer cell line MDA-MB-231 and human prostate cancer cell line PC-3. Among them, compounds ZYR-FA-4, ZYR-FA-7, and ZYR-FA-13 showed significant anti-proliferative activity, inhibiting the proliferation of both tumor cell lines by IC50. 50 Approximately 5 μM.

[0131] Table 1. Effects of the compounds in the examples on the proliferation of MDA-MB-231 and PC-3 cells.

[0132]

[0133]

[0134]

[0135] Because compounds ZYR-FA-4, ZYR-FA-7, and ZYR-FA-13 exhibit significant anti-proliferative activity, the proliferation-inhibiting activities of these three representative compounds in various tumor cell lines were investigated in the embodiments of this invention, including lung cancer cell lines H1975, NCI-H460, squamous cell carcinoma cell line A431, pancreatic cancer cell line HPAF-II, gastric cancer cell lines NUGC-3 and KATOIII, colon cancer cell line HCT116, diffuse large B-cell lymphoma cell lines DOHH2 and SUDHL-4, chronic myeloid leukemia cell line K562, and acute myeloid leukemia cell line MV-4-11. Cells were grown in DMEM or RPMI 1640 medium containing 10% fetal bovine serum and 1% penicillin-dextrose antibodies at 37°C and 5% CO2.

[0136] Experimental methods

[0137] The proliferation inhibition assay for adherent cells such as H1975, NCI-H460, A431, HPAF-II, NUGC-3, KATO III and HCT116 cells was performed using the SRB method (as above).

[0138] The proliferation inhibition assay for suspension cells such as DOHH2, SUDHL-4, K562, and MV-4-11 was performed using the MTT assay. Details are as follows:

[0139] Take cells in good logarithmic growth phase and use 1×10 4Cells were seeded at a density of [number] cells / well in 96-well plates and incubated at 37°C in a 5% CO2 incubator for 24 hours. Different concentrations of the compound of this invention were added: 100 μM, 30 μM, 10 μM, 3 μM, 1 μM, 300 nM, 100 nM, 30 nM, and 10 nM. An equal volume of DMSO was added to the control group. The plates were incubated for 5 days. MTT reagent was added to each well at 10 μL / well. After reacting in an incubator for 4 hours, 100 μL of triplet solution (4%, w / v) was added to each well. After the crystals dissolved, the absorbance of each well was measured at 570 nm using a microplate reader, and the cell proliferation inhibition rate (%) was calculated according to the following formula.

[0140]

[0141] Statistical analysis was performed on the effects of different concentrations of the compounds of the present invention on the proliferation of the aforementioned tumor cells. GraphPadPrism software was used for IC50 analysis of different compounds. 50 The calculations and test results are shown in Table 2.

[0142] Experimental results

[0143] The results, as shown in Table 2, indicate that the representative compounds in the embodiments of this invention exhibit significant inhibitory activity against various tumor cell lines (lung cancer cell lines H1975, NCI-H460, squamous cell carcinoma cell line A431, pancreatic cancer cell line HPAF-II, gastric cancer cell lines NUGC-3, KATOIII, colon cancer cell line HCT116, diffuse large B-cell lymphoma cell lines DOHH2, SUDHL-4, chronic myeloid leukemia cell line K562, and acute myeloid leukemia cell line MV-4-11), demonstrating broad-spectrum anti-tumor cell proliferation activity.

[0144] Table 2. Effects of the compounds in the examples on the proliferation of various tumor cells.

[0145]

[0146]

[0147] Example 8: Anti-multidrug resistance effect of the compound in the example

[0148] The compound was used to assess the multidrug resistance of tumor cells using K562 chronic leukemia cells and K562 / ADR multidrug-resistant cells. Cells were grown in RPMI 1640 medium containing 10% fetal bovine serum and 1% penicillin-drug antibodies at 37°C and 5% CO2. 8.1 Effect of the compound on the proliferation of K562 / ADR multidrug-resistant cells.

[0149] Experimental methods

[0150] Same as the MTT determination method in Example 7.

[0151] Experimental results

[0152] K562 / ADR cells were constructed by gradually increasing the concentration of doxorubicin (ADR) in parental K562 cells. The IC50 values ​​of cytotoxic drugs and representative compounds ZYR-FA-4 and ZYR-FA-13 in the multidrug-resistant cell line K562 / ADR and the parental K562 cell line were determined using the MTT assay. 50 The test results are detailed in Table 3. The results show that the resistance index (RI) of K562 / ADR cells to multiple cytotoxic drugs, paclitaxel, vinorelbine, and doxorubicin, were >6000, 230.8, and 106.4 times, respectively. In the embodiments of this invention, the representative compounds ZYR-FA-4 and ZYR-FA-13 maintained IC50 values ​​close to those of the parental cells in the K562 / ADR multidrug-resistant cell line. 50 Values ​​are shown in Table 3.

[0153] Table 3. Effects of the compounds in the examples on the proliferation of K562 / ADR multidrug-resistant cells.

[0154]

[0155] 8.2 Effects of the compound on MDR1 expression in multidrug-resistant K562 / ADR cells

[0156] Experimental methods

[0157] K562 / ADR cells, a type of chronic leukemia cell line in good logarithmic growth phase, were seeded in six-well plates at a density of 2 × 10⁶ cells per well. 5The cells were incubated at 37°C in a 5% CO2 incubator for 24 hours. After incubation for 24 hours with different concentrations (3 and 10 μM) of compounds ZYR-FA-4 and ZYR-FA-13, the cells were lysed with 1×SDS gel loading buffer (50 mM Tris-HCl (pH 6.8), 100 mM DTT, 2% SDS, 10% glycerol, and 0.1% bromophenol blue). Cell lysates were denatured by heating in a boiling water bath and subjected to SDS-PAGE electrophoresis. After electrophoresis, proteins were transferred to a PVDF membrane using a wet transfer system. The PVDF membrane was then blocked at room temperature in blocking buffer (5% skim milk powder diluted in TBST). After blocking, the membrane was incubated overnight at 4°C with the corresponding anti-MDR1 primary antibody (1:1000, #13342, CST) and anti-GAPDH primary antibody (1:2000, #5174, CST). After washing, the bands were immersed in secondary antibody buffer according to the primary antibody species (mouse or rabbit), and incubated on a shaker at room temperature for 1 hour. Finally, the membrane was developed using Immobilon Western HRP Substrate Luminal Reagent and photographed using a Western Blot imaging system.

[0158] Experimental results

[0159] like Figure 1 As shown, compared with K562 cells, K562 / ADR cells exhibited high expression of MDR1, as demonstrated by compound ZYR-FA-4 in the example. Figure 1 A) and ZYR-FA-13 ( Figure 1 B) Significantly inhibited the multidrug resistance protein MDR1 in K562 / ADR cells, with the inhibitory effect being concentration-dependent.

[0160] The above embodiments are merely illustrative of the principles and effects of the present invention and are not intended to limit the invention. Any person skilled in the art can modify or alter the above embodiments without departing from the spirit and scope of the present invention. Therefore, all equivalent modifications or alterations made by those skilled in the art without departing from the spirit and technical concept disclosed in the present invention should still be covered by the claims of the present invention.

Claims

1. A carborane-containing isoflavone derivative, characterized in that, The compound represented by formula (Ⅰ): Among them, R 1 and R 2 Each is independently selected from hydrogen atom, hydroxyl group, benzyloxy group, methoxy group, sulfonyl fluoroxy group; X is selected from carbonyl group, methylene group, CH-OH group; XY is selected from carbon-carbon single bonds or carbon-carbon double bonds; YZ is selected from carbon-carbon single bonds or carbon-carbon double bonds.

2. The carborane-containing isoflavone derivative as described in claim 1, characterized in that, The compound represented by formula (Ⅰ) is selected from the following group: Group 1: Compounds shown in Formula D Among them, R 3 and R 4 Each is independently selected from hydrogen atoms, methoxy groups, and benzyloxy groups; Group 2: Compounds shown in Formula E Among them, R 8 Selected from hydroxyl or methoxy, R 9 Selected from hydrogen atoms or hydroxyl groups; Group 3: Compounds shown in Formula F Among them, R 10 Selected from methoxy or hydroxyl groups; Group 4: Compounds shown in Formula G Among them, R 7 Selected from methoxy or sulfonyl fluoroxy; Q is selected from carbonyl or methylene; QT is selected from carbon-carbon single bond or carbon-carbon double bond; TL is selected from carbon-carbon single bond or carbon-carbon double bond.

3. The carborane-containing isoflavone derivative as described in claim 2, characterized in that, The compound shown in formula D is selected from 4. The carborane-containing isoflavone derivative as described in claim 2, characterized in that, The compound shown in Formula E is selected from 5. The carborane-containing isoflavone derivative as described in claim 2, characterized in that, The compound shown in formula F is selected from 6. The carborane-containing isoflavone derivative as described in claim 2, characterized in that, The compound shown in formula G is selected from 7. A method for preparing the carborane-containing isoflavone derivative as described in claim 3, characterized in that, Includes the following steps: S1. Compound A is coupled with trimethylsilylacetylene in the presence of palladium catalyst and cuprous halide via a Sonogashira coupling reaction to provide compound B. S2. Compound B is subjected to acidic conditions and the action of tetrabutylammonium fluoride to remove the TMS protecting group, in order to provide compound C. S3, compound C, and decaboron-dodecyl diacetonitrile complex were reacted in the presence of ionic solution (Bmim)Cl to give compound D; The reaction formula is as follows: Among them, R 3 and R 4 Each is independently selected from hydrogen atom, methoxy group, benzyloxy group, R 5 Selected from Br or I.

8. The method for preparing carborane-containing isoflavone derivatives as described in claim 4, characterized in that, Includes the following steps: Compound D of claim 2 is added dropwise with boron tribromide solution under ice bath conditions. After the reaction is completed, saturated ammonium chloride aqueous solution is added to quench the reaction to provide compound E. The reaction formula is as follows: Among them, R 3 It is a methoxy group, R 4 Selected from hydrogen atoms or methoxy groups, R 8 Selected from hydroxyl or methoxy, R 9 Selected from hydrogen atoms or hydroxyl groups.

9. The method for preparing carborane-containing isoflavone derivatives as described in claim 5, characterized in that, Includes the following steps: The compound of formula E in claim 2 is reacted with a borohydride to provide compound of formula F; The reaction formula is as follows: Among them, R 8 Selected from methoxy or hydroxyl, R 9 For hydrogen atoms, R 10 Selected from methoxy or hydroxyl groups.

10. The method for preparing carborane-containing isoflavone derivatives as described in claim 6, characterized in that, Includes any of the following steps: (a) The R in claim 2 10 Compounds of formula F with a methoxy group undergo oxidation in the presence of manganese dioxide to provide R. 7 The compound of formula G has a methoxy group, a carbonyl group, a carbon-carbon single bond, and a carbon-carbon single bond. (b) The R in claim 2 10 The methoxylated compound of formula F reacts with polyphosphoric acid, and after the reaction is complete, it is quenched with water and purified to provide R. 7 Compound G has the following structure: Q is methoxy, Q is methylene, QT is carbon-carbon double bond, and TL is carbon-carbon single bond. (c) The R in claim 2 10 The methoxylated compound of formula F reacts with polyphosphoric acid, and after the reaction is complete, it is quenched with water and purified to provide R. 7 Compound G is a compound with a methoxy group, Q is a methylene group, QT is a carbon-carbon double bond, and TL is a carbon-carbon single bond; the resulting R... 7 Compound G, consisting of a methoxy group, Q being a methylene group, QT being a carbon-carbon double bond, and TL being a carbon-carbon single bond, undergoes an addition reaction with hydrogen in the presence of a palladium-on-carbon catalyst to provide R. 7 The compound of formula G has methoxy group, Q is methylene group, QT is carbon-carbon single bond, and TL is carbon-carbon single bond. (d) R in claim 2 8 For hydroxyl group, R 9 Compound E, with hydrogen atoms, reacts with 1-(fluorosulfonyl)-2,3-dimethyl-1H-imidazolium-3-onium trifluoromethanesulfonate in the presence of a base to provide R. 7 It is a compound of formula G, where Q is a sulfonyl fluoride group, Q is a carbonyl group, QT is a carbon-carbon single bond, and TL is a carbon-carbon double bond.

11. Use of the carborane isoflavone derivatives or pharmaceutically acceptable salts thereof as described in any one of claims 1 to 6 in the preparation of antitumor drugs.

12. Use of the carborane isoflavone derivatives or pharmaceutically acceptable salts thereof as described in any one of claims 1 to 6 in the preparation of anti-multidrug resistant tumor drugs.

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