Chalcone and flavanone sulfamide derivatives, synthesis method and application thereof

Abstract

This invention discloses 2'-hydroxy-4,4',6'-trimethoxychalcone and 5,7,4'-trimethoxyflavanone sulfonamide derivatives, their synthesis methods and applications, belonging to the field of pharmaceutical technology, and involving two series of compounds: 3-sulfonamido-2'-hydroxy-4,4',6'-trimethoxychalcone and 8,3'-disulfonamido-5,7,4'-trimethoxyflavanone, and their preparation methods. Starting with 2-hydroxy-4,6-dimethoxyacetophenone and p-anisaldehyde or 4-methoxy-3-nitrobenzaldehyde, 2'-hydroxy-4,4',6'-trimethoxychalcone or 3-amino-2'-hydroxy-4,4',6'-trimethoxychalcone intermediates were synthesized via the Claisen-Schmidt reaction. Then, through sulfonation and intramolecular Michael addition reactions, 2'-hydroxy-4,4',6'-trimethoxychalcone and 5,7,4'-trimethoxyflavanone sulfonamide derivatives were synthesized. The preparation method is simple, mild, and yields high results. In vitro and in vivo activity tests show that the derivatives disclosed in this invention have significant in vitro inhibitory activity against gastric cancer, esophageal cancer, colorectal cancer, and lung cancer cell lines, acting as inhibitors of the protein YBX1 / RPN1. Clinically, they can be applied to targeted therapy for cancers such as gastric cancer, esophageal cancer, colorectal cancer, and lung cancer.

Description

Technical Field

[0001] This invention belongs to the field of pharmaceutical technology, specifically relating to 2'-hydroxy-4,4',6'-trimethoxychalcone and 5,7,4'-trimethoxyflavanone sulfonamide derivatives, their synthesis methods and applications. Background Technology

[0002] Drug resistance in cancer and the low specificity of anticancer drugs have always been major challenges in cancer control and treatment, urgently requiring the development of novel and highly effective anticancer drugs. Chalcones are widely distributed in fruits, vegetables, spices, tea, and soybeans. The α,β-unsaturated ketones with three carbon atoms are the active functional groups and are precursors to flavonoids and isoflavones (Rajendran et al. Pharmaceuticals 2022, 15:1250). Chalcones have potential activity against drug-sensitive and drug-resistant cancers, acting on various drug targets, and are valuable for developing novel anticancer drugs. Derivatives hybridized with other pharmacophores have shown potential to inhibit the growth of drug-resistant cancers and improve specificity; therefore, this is an effective strategy for developing novel anticancer drugs (Gao et al. Med Res Rev. 2020:1-36). Derivatives hybridized with the sulfonamide at the C-5' position of chalcone show GI activity against the K-562 leukemia cell line and the LOX-IMVI melanoma cell line. 50 The GI of glioma U251 cell lines was induced by introducing sulfonamide pyrimidinone hybridization derivatives at concentrations of 0.57 and 1.28 μM, respectively. 50 The concentration was 1.58 μM (Castaño et al. Eur JMed Chem. 2019, 176: 50-60). Recently, chalcone derivatives targeting mitochondria (CN 115594715 A) and chalcone derivatives for esophageal cancer (CN 115260038 A) have been granted patents and can be used in the development of anti-tumor drugs.

[0003]

[0004] 2'-Hydroxy-4,4',6'-Trimethoxychalcone is a chalcone compound distributed in *Goniothalamus thwaitesii*, *Goniothalamus thwaitesii*, *Kaempferia angustifolia*, and *Goniothalamus thwaitesii*, and is a precursor of 5,7,4'-trimethoxyflavanone. Recently, we discovered that 2'-hydroxy-4,4',6'-trimethoxychalcone exhibits in vitro inhibitory activity against the proliferation of esophageal cancer, colorectal cancer, gastric cancer, and lung cancer, with an IC50 value... 50With a molecular weight range of 15.39 ~ 28.62 μM, it is an effective inhibitor of YBX1 / RPN1 protein without significant toxicity. Sulfonamide structures possess diverse pharmacological properties and serve as important scaffolds for drug development. To enhance the anticancer activity of 2'-hydroxy-4,4',6'-trimethoxychalcone and overcome the aforementioned bottlenecks in cancer treatment, we consider introducing sulfonamide at the 3' position. Furthermore, we propose simultaneously modifying the structure by introducing sulfonamide at both the 8' and 3' positions to further improve its anticancer activity.

[0005] The 2'-hydroxy-4,4',6'-trimethoxychalcone and 5,7,4'-trimethoxyflavanone sulfonamide derivatives disclosed in this invention are inhibitors of the target protein YBX1 / RPN1. They have potential application value in developing into novel drugs for the treatment of esophageal cancer, gastric cancer, colorectal cancer, lung cancer and other cancers. They are of great significance for the development of drugs with independent intellectual property rights. No relevant literature reports have been found so far. Summary of the Invention

[0006] The purpose of this invention is to provide 2'-hydroxy-4,4',6'-trimethoxychalcone and 5,7,4'-trimethoxyflavanone sulfonamide derivatives, their synthesis methods and applications.

[0007] To achieve the objective of this invention, the technical solution is as follows:

[0008] 2'-hydroxy-4,4',6'-trimethoxychalcone and 5,7,4'-trimethoxyflavanone sulfonamide derivatives have the following general structural formulas:

[0009] R1, R2, and R3 are each independently selected from ethyl, cyclopropyl, 2,4-difluorophenyl, 4-fluorophenyl, 4-chlorophenyl, 2,3-dichlorophenyl, 3-bromophenyl, phenyl, 4-methoxyphenyl, 4-methylphenyl, 4-bromophenyl, 4-nitrophenyl, 4-cyanophenyl, 3-bromophenyl, 3-methoxyphenyl, homoserine lactone, 3-chlorophenyl, 2-fluorophenyl, 3,4,5-trimethoxyphenyl, 2-chlorophenyl, 2-methylphenyl, 3-methylphenyl, 3-chlorophenyl, 2-fluorophenyl, and 3-fluorophenyl.

[0010] Preferably, the compound has the following structure:

[0011]

[0012]

[0013] This invention provides a method for preparing 2'-hydroxy-4,4',6'-trimethoxychalcone and 5,7,4'-trimethoxyflavanone sulfonamide derivatives. The reaction route of general formula I is as follows:

[0014]

[0015] The synthesis process of general formula I is as follows:

[0016] (1) Dissolve 2-hydroxy-4,6-dimethoxyacetophenone (compound 1) and 4-methoxy-3-nitrobenzaldehyde (compound 2a) in the reaction solvent, add alkali, heat to reflux, and a Claisen-Schmidt reaction occurs. After the reaction is complete, cool to room temperature, pour the reaction solution into ice water, adjust the pH of the system to 5-6 with dilute hydrochloric acid, precipitate out, filter, wash, and obtain 3-nitro-2'-hydroxy-4,4',6'-trimethoxychalcone (compound 3a), which can be directly used in the next step of the reaction;

[0017] (2) Compound 3a was dissolved in the reaction solvent, and reducing metal powder was added. Acid was added and refluxed to reduce the nitro group to an amino group. The mixture was purified by silica gel column chromatography to obtain 3-amino-2'-hydroxy-4,4',6'-trimethoxychalcone (compound 4).

[0018] (3) Compound 4 was dissolved in the reaction solvent, and an acylation reaction catalyst and a base were added. The mixture was then reacted with commercially available sulfonyl chloride (compound 5) to synthesize a series of derivatives 6 of general formula I.

[0019] Preferably, in step (1) of synthesizing the derivative of general formula I, the molar ratio of Claisen-Schmidt reaction compound 1, compound 2a, and base is 1:(1.2~1.4):(2~5), and the solvent used is methanol, ethanol, tert-butanol, or acetonitrile; the base used is one or a mixture of two or more of potassium carbonate, sodium carbonate, sodium hydroxide, lithium hydroxide, and sodium hydride; and the concentration of dilute hydrochloric acid is 5~ 10 mol / L; the recrystallization solvent is methanol, ethanol or tert-butanol; in step (2), the molar ratio of compound 4a, reducing metal powder and acid is 1:(5~8):(5~10), the selected reaction solvent is methanol, ethanol, isopropanol or tert-butanol, the reducing metal powder is iron powder, magnesium powder or zinc powder, and the acid is concentrated hydrochloric acid, concentrated sulfuric acid or glacial acetic acid; in step (3), the molar ratio of compound 4, commercially available sulfonyl chloride (compound 5), catalyst and base is 1:(1.5~2):(1~2):(3~5), the selected reaction solvent is acetone, tetrahydrofuran or 1,4-dioxane, the catalyst is DMAP or DCC, and the base is triethylamine, pyridine or DBU.

[0020] The reaction route for formula II is as follows:

[0021]

[0022] The synthesis process of Formula II is as follows:

[0023] (1) Dissolve 2-hydroxy-4,6-dimethoxyacetophenone (compound 1) and p-anisaldehyde (compound 2b) in a reaction solvent, add alkali, heat to 110 ± 10℃ to carry out the Claisen-Schmidt reaction. After the reaction is complete, cool to room temperature, pour the reaction solution into ice water, adjust the pH of the system to 5 ~ 6 with dilute hydrochloric acid, precipitate, filter, wash, recrystallize to obtain 2'-hydroxy-4,4',6'-trimethoxychalcone (compound 3b);

[0024] (2) At 0 ± 5℃, compound 3b was added to the sulfonating agent. After the addition was complete, the mixture was moved to room temperature to sulfonate compound 3b. After the reaction was complete, the reaction was quenched by ice water, and a precipitate was formed. The precipitate was filtered, washed with pure water, and a solid was obtained. The prepared sulfonyl chloride (compound 7) can be used directly in the next step of the reaction.

[0025] (3) Dissolve compound 7 in the reaction solvent, add commercially available amine compound 8, stir at room temperature until the reaction is complete, evaporate to dryness, dissolve the residue in ethyl acetate, wash with hydrochloric acid solution until neutral, wash with saturated brine, dry with anhydrous sodium sulfate, filter, concentrate, and purify by silica gel column chromatography to obtain a series of derivatives 9 of general formula II.

[0026] Preferably, in step (1) of synthesizing general formula II derivatives, the molar ratio of Claisen-Schmidt reaction compound 1, compound 2a or compound 2b, and base is 1:(1.2~1.4):(2~5), and the solvent used is methanol, ethanol, tert-butanol, or acetonitrile; the base used is one or a mixture of two or more of potassium carbonate, sodium carbonate, sodium hydroxide, lithium hydroxide, and sodium hydride; the concentration of dilute hydrochloric acid is 5~10 mol / L;

[0027] In step (2), the molar ratio of compound 3b to chlorosulfonic acid is 1:(17~18).

[0028] In step (3), the molar ratio of compound 7 to commercially available amine (compound 8) is 1:(1.5~2), and the solvent used is acetone, tetrahydrofuran or 1,4-dioxane;

[0029] The preparation methods of the 2'-hydroxy-4,4',6'-trimethoxychalcone and 5,7,4'-trimethoxyflavanone sulfonamide derivatives provided by this invention are simple and have potential application value. They are YBX1 / RPN1 protein-targeting inhibitors. In particular, compounds 6i, 9e, 9i, and 9l have good inhibitory activity against gastric cancer, esophageal cancer, colorectal cancer, and lung cancer cells at concentrations of 1-8 μM. Clinically, they can be developed into drugs for targeted therapy of gastric cancer, esophageal cancer, colorectal cancer, lung cancer, and other cancers, which can improve patients' survival rate and quality of life. Attached Figure Description

[0030] Figure 1 NMR spectra of compound 3b and derivative 9a of this invention 1 H Comparison of NMR spectra;

[0031] Figure 2 The NMR of derivative 9a of this invention 1 H 1 HCOSY diagram;

[0032] Figure 3 The NMR HSQC spectrum of derivative 9a of this invention;

[0033] Figure 4 The NMR HMBC spectrum of derivative 9a of this invention;

[0034] Figure 5 The NMR spectrum of derivative 9j of this invention is DEPT135.

[0035] Figure 6 The derivative 9j of this invention is a nuclear magnetic resonance image. 1 H 1 HCOSY diagram;

[0036] Figure 7 The NMR HSQC spectrum of derivative 9j of this invention;

[0037] Figure 8 The NMR HMBC spectrum of derivative 9j of this invention;

[0038] Figure 9 The derivatives 9e, 9i, 9l, 6i and 3b of this invention can inhibit the proliferation of esophageal squamous cell carcinoma KYSE450, gastric cancer cells HGC27 and colorectal cancer cells HCT116 at concentrations ranging from 1 to 8 μM.

[0039] Figure 10 The derivatives 9e, 9i, 9l, 6i and 3b of this invention can inhibit the colony formation of esophageal squamous cell carcinoma KYSE450, gastric cancer cells HGC27 and colorectal cancer cells HCT116 at a concentration range of 1 ~ 8 μM.

[0040] Figure 11 The tumor therapeutic effects of derivatives 9e, 9i, 9l, 6i, and 3b of this invention on a mouse model of human gastric cancer HGC27 cell xenografts are shown in the following figures: A is a graph showing the trend of tumor volume changes during drug administration; B is the tumor weight measured after the mice were sacrificed at the end of the experiment; C is a tumor image of the control group and the drug administration group; D is a graph showing the trend of mouse body weight changes during drug administration; E, F, and G are the weights of the liver, spleen, and kidneys of the mice at the end of the experiment.

[0041] Figure 12The derivatives 9e, 9i, 9l, 6i, and 3b of this invention target YBX1 and RPN1: where A is the Venn analysis diagram of the mass spectrometry results; B is the Metascape protein interaction network diagram of the mass spectrometry results; C and D are the results of the hydrogen bromide-activated Sepharose 4B protein pull-down experiment in human gastric cancer HGC27 cells (target proteins YBX1 and RPN1).

[0042] Note: Figures 9 to 11 In the study, compared with the control group, * indicates P < 0.05; ** indicates P < 0.01; *** indicates P < 0.001.

[0043] Materials and Methods

[0044] 1. Materials

[0045] 1.1 Tumor cell lines

[0046] The esophageal cancer cell lines, gastric cancer cell lines, colorectal cancer cell lines, and lung cancer cell lines used in this invention are from the Sino-US (Henan) Hormel Cancer Institute.

[0047] 1.2 Reagents

[0048] Penicillin: North China Pharmaceutical Co., Ltd.;

[0049] Streptomycin: Shandong Lukang Pharmaceutical Co., Ltd.;

[0050] RPMI-1640 culture medium: Biological Industries, Israel;

[0051] 0.25% Pancreatic Enzyme: Shanghai Beyotime Biotechnology Co., Ltd.;

[0052] Serum-free cell cryopreservation solution: Suzhou Xinsaimei Biotechnology Co., Ltd.;

[0053] PBS powder: Beijing Solarbio Science & Technology Co., Ltd.

[0054] MTT powder: Beijing Solarbio Technology Co., Ltd.;

[0055] Crystal violet powder: Tianjin Kemeo Chemical Reagent Co., Ltd.;

[0056] DMSO: Tianjin Kemeo Chemical Reagent Co., Ltd.

[0057] Fetal bovine serum: BI, Inc., USA;

[0058] CNBr-activated Sepharose 4B beads: Cytiva Corporation, Japan;

[0059] RPN1 antibody: Abcam, UK;

[0060] YBX1 antibody: Proteintech, USA;

[0061] 0.4% Sodium Pentobarbital: Sinopharm Chemical Reagent Co., Ltd.;

[0062] 500ml bottle of physiological saline: Chenxin Pharmaceutical Co., Ltd.

[0063] 1.3 Instruments and Consumables:

[0064] 1.5 ml centrifuge tubes: Axygen Corporation, USA;

[0065] 15 ml centrifuge tubes: Corning Corporation, USA;

[0066] 96-well cell culture plate: Wuxi NiceBiotechnology Co., Ltd.;

[0067] 6-well cell culture plate: Wuxi NiceBiotechnology Co., Ltd.;

[0068] 10 cm cell culture dish: Wuxi NiceBiotechnology Co., Ltd.;

[0069] 15 cm cell culture dish: Thermo Fisher Scientific, USA;

[0070] Disposable pipettes: Guangzhou Jetech Biofiltration Co., Ltd.;

[0071] Ultra-sensitive multi-functional imager: Cytiva Corporation, Japan;

[0072] Pipettes: Eppendorf, Germany;

[0073] Dry CO2 incubator: Shanghai Yiheng Scientific Instruments Co., Ltd.;

[0074] High-speed low-temperature centrifuge: Eppendorf GmbH, Germany;

[0075] Vacuum suction pump: Haimen Qilin Bell Instrument Manufacturing Co., Ltd.;

[0076] Snowflake ice maker: SANYO Corporation, Japan;

[0077] Mili-Q water purifier: Millipore, USA;

[0078] Thermo clean bench; ophthalmic scissors, surgical forceps, scalpels, dissolving needles, syringes.

[0079] 1.4 Laboratory Animals

[0080] Five-week-old BALB / c-nu mice were purchased from Henan Xincheng Youkang Biotechnology Co., Ltd., and housed in a constant temperature and pressure environment with a 12-hour day-night cycle. Mice were only considered for experiments when their weight reached approximately 18–20 g. Mouse feed was purchased from Beijing Huafu Biotechnology Co., Ltd. The experimental animals were housed in the animal facilities of the School of Life Sciences, Zhengzhou University, Henan Province, under conditions of constant temperature (25–27°C), constant humidity (45%–50%), fresh air, and a dust-free, sterile, and pathogen-free (SPF-grade) environment. Animals were initially placed in organic plastic boxes (Suzhou Fengshi Experimental Animal Equipment Co., Ltd.) and then placed in an IVC system. Aseptically treated feed was provided for free access. High-temperature sterilized bedding was changed every three days, and cages and drinking water were ultraviolet-sterilized every three days. Sterile distilled water was provided as drinking water. Strict aseptic techniques were followed when changing feeding supplies. The experimental animals were housed under a 12 / 12-hour light / dark cycle, with free access to food and water, and the room temperature was controlled at 25°C. Detailed Implementation

[0081] The present invention will be further described below with reference to the embodiments, in which the solvent used in column chromatography is a volume ratio.

[0082] Example 1: Preparation of compounds 4, 6a-6m, and 9a-9t

[0083] (1) Preparation of compound 3a

[0084] Compound 1 (1.08 g, 5.25 mmol, 1.0 equiv) was dissolved in methanol (30 mL) and potassium hydroxide (703 mg, 10.5 mmol, 2.0 equiv) was added. Compound 2a (962 mg, 5.31 mmol, 1.01 equiv) was added to the reaction solution, and the mixture was refluxed for 4 hours until compound 1 was completely consumed. After cooling to room temperature, the reaction solution was poured into an ice-water mixture and the pH was adjusted to 5-6 with dilute hydrochloric acid, resulting in the formation of a large amount of yellow solid. The solid was filtered, washed, and dried to obtain 1.69 g of compound 3a, with a yield of 89.9%, which could be used directly in the next reaction.

[0085] (2) Preparation of compound 4

[0086] Compound 3a (552.4 mg, 1.54 mmol, 1.0 equiv) was mixed thoroughly with ethanol (10 mL), and concentrated hydrochloric acid (2 mL) and reduced iron powder (860.9 mg, 15.4 mmol, 10.0 equiv) were added. The reaction mixture was refluxed for 4 hours. After the reaction was complete, diatomaceous earth was filtered while hot and washed with ethyl acetate. The filtrate was adjusted to pH 10–11 with sodium hydroxide solution (2 mol / L), extracted with ethyl acetate, and the organic phase was separated, washed with saturated brine, and dried over anhydrous sodium sulfate. After filtration and rotary evaporation, the brown residue was purified by silica gel column chromatography (DCM / MeOH = 80:1). 358.9 mg of yellow solid was obtained, yield: 70.1%. mp: 119.4–123.5 °C. 1 H NMR (400 MHz, CDCl3) δ 14.42 (s, 1H), 7.73 (d, J = 1.2 Hz, 2H), 7.01 (dd, J = 8.2, 2.1 Hz, 1H), 6.99 (d, J = 2.0 Hz, 1H), 6.80 (d, J = 8.2 Hz,1H), 6.10 (d, J = 2.4 Hz, 1H), 5.96 (d, J = 2.4 Hz, 1H), 3.91 (s, 3H), 3.90(s, 3H), 3.88 (br s, 2H), 3.83 (s, 3H). 13 C NMR (101 MHz, CDCl3) δ 187.92,163.57, 161.19, 157.71, 144.67, 138.52, 131.69, 123.91, 120.17, 116.06,108.86, 105.50, 101.66, 89.05, 86.48, 51.13, 50.86, 50.82. HRMS (ESI) [M+H] + calcd for C 18 H 20 NO5: 330.1341, found:330.1343.

[0087] (4) Preparation of compounds (6a-6m) represented by general formula 6

[0088] Compound 4 (51.8 mg, 0.16 mmol, 1.0 equiv) was dissolved in tetrahydrofuran, and DMAP (39.1 mg, 0.32 mmol, 2 equiv) and pyridine (130 μL, 1.6 mmol, 10.0 equiv) were added. Commercially available sulfonyl chloride compound 5 (0.32 mmol, 2.0 equiv) was added to the reaction mixture, and the mixture was reacted overnight at room temperature. The reaction mixture was poured into dilute hydrochloric acid solution, extracted with ethyl acetate, and the organic phases were combined, dried over anhydrous sodium sulfate, and evaporated to dryness. The brown residue was purified by silica gel column chromatography (DCM / MeOH = 60:1) to give compounds 6a-6m.

[0089] Compound 6a: pale yellow solid, yield: 77.4%. 1 H NMR (400 MHz, CDCl3) δ 14.40 (br s,1H), 7.98 (d, J = 1.7 Hz, 1H), 7.92 (d, J = 15.5 Hz, 1H), 7.71 (d, J = 15.5Hz, 1H), 7.29 (dd, J = 8.5, 1.8 Hz, 1H), 6.92 (d, J = 8.4 Hz, 1H), 6.83 (s,1H), 6.10 (d, J = 2.2 Hz, 1H), 5.97 (d, J = 2.2 Hz, 1H), 3.95 (s, 3H), 3.94(s, 3H), 3.84 (s, 3H), 3.11 (q, J = 7.4 Hz, 2H), 1.35 (t, J = 7.4 Hz, 3H). 13 CNMR (101 MHz, CDCl3) δ 192.40, 168.54, 166.26, 162.64, 150.21, 141.27,129.29, 126.98, 126.95, 126.76, 117.60, 110.73, 106.25, 93.78, 91.24, 56.08,55.79, 55.60, 45.68, 8.19. HRMS (ESI) [M+H] + calcd for C 20 H 24 NO7S: 422.1273, found: 422.1277.

[0090] Compound 6b: pale yellow solid, yield: 76.7%. 1H NMR (400 MHz, DMSO-d6) δ 7.11 (d, J= 16.0 Hz, 1H), 6.92 (d, J = 1.3 Hz, 1H), 6.88 – 6.80 (m, 2H), 6.75 – 6.67(m, 2H), 6.63 (d, J = 2.0 Hz, 1H), 4.86 (br s, 2H), 3.86 (s, 3H), 3.81 (s,3H), 3.78 (s, 3H), 2.99 – 2.89 (m, 1H), 1.17 – 1.09 (m, 2H), 1.00 (dt, J =8.8, 4.5 Hz, 2H). 13 C NMR (101 MHz, DMSO-d6) δ 190.92, 161.68, 158.85, 149.58,147.50, 146.56, 138.54, 127.25, 125.58, 119.68, 116.76, 112.46, 110.92,100.23, 97.93, 56.78, 56.34, 55.92, 28.63, 6.66. HRMS (ESI) [M+H] + , calcd forC 21 H 24 NO7S: 434.1273, found: 434.1271.

[0091] Compound 6c: pale yellow solid, yield: 82.4%. 1 H NMR (400 MHz, CDCl3) δ 14.36 (s,1H), 7.91 – 7.79 (m, 3H), 7.65 (d, J = 15.6 Hz, 1H), 7.34 (s, 1H), 7.24 (dd,J = 8.5, 2.0 Hz, 1H), 6.90 (t, J = 8.3 Hz, 2H), 6.79 (d, J = 8.5 Hz, 1H), 6.11 (d, J = 2.3 Hz, 1H), 5.99 (d, J = 2.4 Hz, 1H), 3.98 (s, 3H), 3.85 (s,3H), 3.81 (s, 3H). 13 C NMR (101 MHz, CDCl3) δ 192.39, 168.51, 166.03 (dd, J C,F= 256.7, 11.6 Hz), 166.28, 162.60, 159.82 (dd, J C,F = 257.4, 12.9 Hz), 150.94,141.26, 132.62(d, J C,F = 10.5 Hz), 128.96, 127.27, 126.64, 125.69, 123.36 (dd,J C,F = 13.8, 3.9 Hz), 119.36, 111.77 (dd, J C,F = 21.9, 3.7 Hz), 110.69, 106.27,105.52 (t, J C,F = 25.0 Hz), 93.82, 91.31, 55.97, 55.85, 55.62. HRMS (ESI) [M+H] + calcd for C 24 H 22 F2NO7S: 506.1085, found:506.1077.

[0092] Compound 6d: pale yellow solid, yield: 84.6%. 1 H NMR (400 MHz, CDCl3) δ 14.38 (br s,1H), 7.96 (d, J = 1.6 Hz, 1H), 7.91 (d, J = 15.5 Hz, 1H), 7.77 (dd, J = 8.7,5.0 Hz, 2H), 7.69 (d, J = 15.6 Hz, 1H), 7.24 (d, J = 1.7 Hz, 1H), 7.08 (t, J= 8.5 Hz, 2H), 6.99 (s, 1H), 6.76 (d, J = 8.4 Hz, 1H), 6.12 (d, J = 2.1 Hz,1H), 5.99 (d, J = 2.1 Hz, 1H), 3.98 (s, 3H), 3.85 (s, 3H), 3.71 (s, 3H). 13 CNMR (101 MHz, CDCl3) δ 192.41, 168.53, 166.29, 165.26 (d, J C,F = 253.9 Hz),162.62, 151.01, 141.24, 135.08 (d, J C,F= 3.3 Hz), 129.88 (d, J C,F = 9.4 Hz),129.06, 127.49, 126.70, 126.15, 119.75, 116.14 (d, J C,F = 22.6 Hz), 110.69,106.28, 93.82, 91.31, 55.89, 55.87, 55.62. HRMS (ESI) [M+H] + , calcd forC 24 H 23 FNO7S: 488.1179, found: 448.1185.

[0093] Compound 6e: pale yellow solid, yield: 75.1%. 1 H NMR (400 MHz, CDCl3) δ 14.27 (br s,1H), 7.92 – 7.89 (m, 3H), 7.69 (d, J = 11.1 Hz, 1H), 7.67 – 7.63 (m, 1H),7.54 – 7.51 (m, 3H), 7.35 (d, J = 2.1 Hz, 1H), 6.90 (d, J = 8.7 Hz, 1H), 6.12(d, J = 2.4 Hz, 1H), 5.97 (d, J = 2.3 Hz, 1H), 3.86 (s, 3H), 3.85 (s, 3H), 3.50 (s, 3H). 13 C NMR (101 MHz, CDCl3) δ 192.19, 168.48, 166.35, 162.47,158.63, 140.70, 140.16, 138.15, 132.53, 130.33, 129.06, 127.08, 123.00,112.38, 106.26, 93.85, 91.37, 55.83, 55.66, 55.53. HRMS (ESI) [M+H] + , calcdfor C 24 H 23 ClNO7S: 504.0884, found:504.0892.

[0094] Compound 6f: pale yellow solid, yield: 67.3%. 1H NMR (400 MHz, CDCl3) δ 14.36 (br s,1H), 7.95 (dd, J = 7.9, 1.3 Hz, 1H), 7.86 (d, J = 15.2 Hz, 1H), 7.85 (d, J =1.9 Hz, 1H), 7.69 – 7.58 (m, 3H), 7.29 – 7.20 (m, 2H), 6.78 (d, J = 8.5 Hz,1H), 6.11 (d, J = 2.3 Hz, 1H), 5.99 (d, J = 2.3 Hz, 1H), 3.99 (s, 3H), 3.85(s, 3H), 3.80 (s, 3H). 13 C NMR (101 MHz, CDCl3) δ 192.38, 168.51, 166.27,162.58, 150.91, 141.33, 138.42, 135.54, 134.76, 130.36, 130.12, 128.88,127.18, 127.09, 126.55, 125.66, 119.20, 110.74, 106.26, 93.81, 91.32, 55.98,55.87, 55.62. HRMS (ESI) [M+H] + calcd for C 24 H 22 Cl2NO7S: 538.0494, found:538.0499.

[0095] Compound 6g: pale yellow solid, yield: 86.4%. 1 H NMR (400 MHz, CDCl3) δ 14.37 (s,1H), 7.92 (dd, J = 10.6, 8.7 Hz, 3H), 7.73 – 7.60 (m, 3H), 7.30 – 7.24 (m,2H), 7.00 (s, 1H), 6.77 (d, J = 8.5 Hz, 1H), 6.11 (d, J = 2.3 Hz, 1H), 5.99 (d, J = 2.3 Hz, 1H), 3.99 (s, 3H), 3.85 (s, 3H), 3.72 (s, 3H). 13C NMR (101MHz, CDCl3) δ 192.43, 168.52, 166.29, 162.63, 151.12, 141.18, 140.83, 136.04,130.32, 130.03, 129.09, 127.83, 126.76, 125.85, 125.75, 122.84, 119.93,110.74, 106.29, 93.80, 91.30, 55.94, 55.89, 55.63. HRMS (ESI) [M+H] + , calcdfor C 24 H 22 BrNO7S: 548.0379, found:548.0386.

[0096] Compound 6h: pale yellow solid, yield: 83.1%. 1 H NMR (400 MHz, DMSO-d6) δ 13.62 (s,1H), 9.71 (s, 1H), 7.74 (d, J = 7.4 Hz, 2H), 7.64 – 7.58 (m, 4H), 7.54 (t, J= 7.5 Hz, 2H), 7.49 (dd, J = 8.5, 1.5 Hz, 1H), 6.98 (d, J = 8.5 Hz, 1H), 6.15 (dd, J = 12.5, 2.0 Hz, 2H), 3.91 (s, 3H), 3.83 (s, 3H), 3.56 (s, 3H). 13 C NMR(101 MHz, DMSO-d6) δ 192.45, 166.25, 166.02, 162.36, 154.61, 142.45, 140.77,133.18, 129.38, 128.69, 127.72, 127.07, 126.45, 125.98, 124.27, 112.59,106.63, 94.42, 91.64, 56.62, 56.21, 56.18. HRMS (ESI) [M+H] + , calcd forC 24 H 24 NO7S: 470.1273, found: 470.1273.

[0097] Compound 6i: pale yellow solid, yield: 83.7%. 1H NMR (400 MHz, CDCl3) δ 14.33 (s,1H), 7.89 (d, J = 1.7 Hz, 1H), 7.85 (d, J = 15.5 Hz, 1H), 7.63 (dd, J = 12.0,9.7 Hz, 3H), 7.14 (dd, J = 8.4, 1.8 Hz, 1H), 6.94 (s, 1H), 6.79 (d, J = 8.9Hz, 2H), 6.68 (d, J = 8.4 Hz, 1H), 6.05 (d, J = 2.2 Hz, 1H), 5.92 (d, J = 2.2Hz, 1H), 3.92 (s, 3H), 3.78 (s, 3H), 3.73 (s, 3H), 3.66 (s, 3H). 13 C NMR (101MHz, CDCl3) δ 191.47, 167.49, 165.23, 162.12, 161.63, 149.74, 140.49, 129.62,128.30, 127.90, 125.97, 125.72, 125.48, 117.76, 113.03, 109.55, 105.28,92.78, 90.26, 54.89, 54.86, 54.58, 54.55. HRMS (ESI) [M+H] + , calcd forC 25 H 26 NO8S: 500.1379, found: 500.1378.

[0098] Compound 6j: pale yellow solid, yield: 67.9%. 1 H NMR (400 MHz, CDCl3) δ 14.40 (s,1H), 7.96 (d, J = 2.0 Hz, 1H), 7.91 (d, J = 15.5 Hz, 1H), 7.67 (dd, J = 11.9,7.9 Hz, 3H), 7.23 – 7.17 (m, 3H), 7.02 (s, 1H), 6.75 (d, J = 8.4 Hz, 1H), 6.12 (d, J = 2.4 Hz, 1H), 5.99 (d, J = 2.4 Hz, 1H), 3.99 (s, 3H), 3.85 (s,3H), 3.72 (s, 3H), 2.36 (s, 3H). 13C NMR (101 MHz, CDCl3) δ 192.50, 168.52,166.26, 162.66, 150.72, 143.93, 141.50, 136.13, 129.53, 128.95, 127.16,127.03, 126.67, 126.53, 118.72, 110.59, 106.31, 93.81, 91.29, 55.90, 55.88,55.61, 21.53. HRMS (ESI) [M+H] + calcd for C 25 H 26 NO7S: 484.1430, found: 484.1433.

[0099] Compound 6k: pale yellow solid, yield: 81.7%. 1 H NMR (400 MHz, CDCl3) δ 14.27 (s,1H), 7.85 – 7.80 (m, 4H), 7.70 – 7.68 (m, 4H), 7.35 (d, J = 2.1 Hz, 1H), 6.89 (d, J = 8.7 Hz, 1H), 6.12 (d, J = 2.3 Hz, 1H), 5.97 (d, J = 2.3 Hz, 1H), 3.86 (s, 3H), 3.85 (s, 3H), 3.49 (s, 3H). 13 C NMR (101 MHz, CDCl3) δ 192.19,168.48, 166.35, 162.48, 158.61, 140.12, 138.70, 132.54, 132.48, 132.06,130.36, 129.32, 129.08, 127.10, 122.97, 112.39, 106.27, 93.86, 91.38, 55.85,55.65, 55.53. HRMS (ESI) [M+H] + calcd for C 24 H 21 BrNO7S: 548.0379, 550.0358, found: 548.0383, 550.0359.

[0100] Compound 6l: pale yellow solid, yield: 68.9%. 1H NMR (400 MHz, DMSO-d6) δ 13.32 (s,1H), 8.51 (d, J = 8.9 Hz, 3H), 8.14 (d, J = 8.9 Hz, 3H), 7.97 (dd, J = 8.8,2.0 Hz, 1H), 7.65 (d, J = 3.6 Hz, 2H), 7.18 (d, J = 8.8 Hz, 1H), 6.17 (d, J =2.2 Hz, 1H), 6.14 (d, J = 2.2 Hz, 1H), 3.86 (s, 3H), 3.83 (s, 3H), 3.41 (s,3H). 13 C NMR (101 MHz, DMSO-d6) δ 192.64, 165.97, 165.70, 162.25, 158.83,151.28, 144.07, 141.02, 133.67, 133.18, 130.57, 128.84, 127.45, 125.13,121.89, 113.98, 106.92, 94.40, 91.60, 56.62, 56.22, 56.18. HRMS (ESI) [M+H] + calcd for C 24 H 23 N2O9S: 515.1124 found: 515.1120.

[0101] Compound 6m: pale yellow solid, yield: 78.5%. 1 H NMR (400 MHz, DMSO-d6) δ 13.62 (s,1H), 9.72 (s, 1H), 7.65 – 7.54 (m, 3H), 7.46 (dd, J = 17.7, 9.3 Hz, 2H), 7.31(d, J = 7.8 Hz, 1H), 7.27 (s, 1H), 7.18 (dd, J = 8.3, 2.3 Hz, 1H), 7.00 (d, J= 8.5 Hz, 1H), 6.17 (d, J = 1.6 Hz, 1H), 6.14 (d, J = 1.8 Hz, 1H), 3.91 (s,3H), 3.83 (s, 3H), 3.75 (s, 3H), 3.61 (s, 3H). 13C NMR (101 MHz, DMSO-d6) δ192.43, 166.26, 166.04, 162.36, 159.66, 154.46, 142.46, 141.98, 130.61,128.62, 127.73, 126.57, 125.97, 123.67, 119.17, 119.13, 112.61, 112.03,106.63, 94.44, 91.64, 56.60, 56.29, 56.17, 55.98. HRMS (ESI) [M+H] + , calcdfor C 25 H 26 NO8S: 500.1379, found: 500.1385.

[0102] (5) Preparation of compound 3b

[0103] In a thick-walled, pressure-resistant bottle, compounds 1 (9.0 g, 0.0459 mol) and 2b (6.9 g, 0.504 mol) were dissolved in 225 mL of methanol. Potassium hydroxide (6.4 g, 0.115 mol) was added, and the mixture was reacted at 110 °C for 18 hours. After the reaction was complete, the mixture was cooled to room temperature, and the reaction solution was poured into ice water. The pH of the system was adjusted to 5-6 with 10 mol / L dilute hydrochloric acid, resulting in the precipitation of a yellow precipitate. The precipitate was filtered, washed with pure water, and recrystallized from 100 mL of ethanol to give 12.2 g of a pale yellow solid, with a yield of 84.7%. mp 112-114 °C; 1 H NMR (600 MHz, CDCl3) δ14.40 (s, 1H), 7.79 (d, J = 2.2 Hz, 2H), 7.56 (d, J = 8.4 Hz, 2H), 6.93 (d, J= 8.4 Hz, 2H), 6.11 (s, 1H), 5.96 (d, J = 1.3 Hz, 1H), 3.92 (s, 3H), 3.85 (s, 3H), 3.83 (s, 3H). 13 C NMR (151 MHz, CDCl3) δ 192.61, 168.38, 166.03, 162.48,161.38, 142.46, 130.11, 128.36, 125.17, 114.38, 106.39, 93.85, 91.25, 55.84,55.57, 55.40. HRMS (ESI) [M+H] + calcd for C 18 H19 O5: 315.1238, found: 315.1232.

[0104] (6) Preparation of compounds (9a-9t) represented by general formula 9

[0105] At 0°C, compound 3b (130 mg, 0.41 mmol, 1.0 equiv) was slowly added to chlorosulfonic acid (1 mL), and the reaction was allowed to proceed to room temperature for 1 hour. The reaction solution was then poured into an ice-water mixture, and a large amount of yellow solid precipitated, which was sulfonyl chloride (compound 8). The solid was filtered, washed with pure water, and immediately proceeded to the next reaction. Compound 8 was dissolved in tetrahydrofuran, and pyridine (2.01 mmol, 5.0 equiv) and commercially available amine compound 9 (0.82 mmol, 2.0 equiv) were added. The mixture was reacted overnight at room temperature. After the reaction was complete, the reaction solution was poured into dilute hydrochloric acid, extracted with ethyl acetate, and the organic phases were combined, dried over anhydrous sodium sulfate, filtered, and the filtrate was evaporated to dryness. The brown residue was subjected to silica gel column chromatography (DCM / MeOH = 50: 1) to give compounds 9a-9t.

[0106] Compound 9a: pale yellow solid, yield: 60.5%. mp 85~89℃; 1 H NMR (400 MHz, CDCl3) δ11.01 (s, 1H), 8.10 (d, J = 2.1 Hz, 1H), 7.70 (dd, J = 8.6, 2.1 Hz, 1H), 7.37(d, J = 16.0 Hz, 1H), 7.01 (d, J = 3.5 Hz, 1H), 6.98 (d, J = 10.9 Hz, 1H),6.07 (s, 1H), 3.97 (d, J = 4.9 Hz, 6H), 3.86 (s, 3H), 3.41 – 3.28 (m, 8H),1.16 (t, J = 7.1 Hz, 6H), 1.11 (t, J = 7.1 Hz, 6H). 13 C NMR (101 MHz, CDCl3) δ192.80, 162.19, 160.93, 158.04, 157.38, 142.99, 133.82, 131.63, 129.74,127.79, 127.30, 112.50, 110.37, 106.32, 87.31, 56.34, 56.19, 56.03, 42.24,41.81, 14.28. HRMS (ESI) [M+H] + calcd for C26 H 37 N2O9S2: 585.1940, found:585.1942.

[0107] Compound 9b: pale yellow solid, yield: 58.9%. 1 H NMR (400 MHz, CDCl3) δ 7.99 (d, J =2.2 Hz, 1H), 7.94 (dd, J = 8.6, 2.2 Hz, 1H), 7.16 (d, J = 8.6 Hz, 1H), 6.21(s, 1H), 5.56 (dd, J = 12.7, 3.3 Hz, 1H), 5.32 (d, J = 2.6 Hz, 2H), 4.05 (d,J = 5.0 Hz, 3H), 4.03 (s, 3H), 4.01 (s, 3H), 3.07 (dd, J = 16.7, 12.7 Hz,1H), 2.96 (dd, J = 16.7, 3.4 Hz, 1H), 2.35 – 2.26 (m, 1H), 2.15 – 2.09 (m,1H), 0.74 – 0.67 (m, 4H), 0.62 – 0.61 (m, 2H), 0.60 – 0.54 (m, 2H). 13 C NMR(101 MHz, CDCl3) δ 187.71, 164.96, 163.99, 162.40, 156.49, 133.06, 130.40,128.16, 127.39, 112.99, 109.38, 106.12, 89.65, 79.10, 56.91, 56.73, 56.50,44.64, 24.83, 24.60, 6.17, 6.12, 6.10. HRMS (ESI) [M+H] + , calcd forC 24 H 29 N2O9S2: 575.1134, found: 575.1125.

[0108] Compound 9c: pale yellow solid, yield: 53.7%. 1H NMR (400 MHz, CDCl3) δ 8.19 (d, J =2.2 Hz, 1H), 8.07 (d, J = 2.2 Hz, 1H), 7.83 (dd, J = 8.6, 2.2 Hz, 1H), 7.72(dd, J = 8.6, 2.2 Hz, 1H), 7.13 (d, J = 8.7 Hz, 1H), 7.10 (d, J = 8.7 Hz,1H), 6.18 (s, 1H), 6.17 (s, 1H), 5.90 (d, J = 3.4 Hz, 1H), 5.86 (d, J = 3.7Hz, 1H), 5.76 (d, J = 3.1 Hz, 1H), 5.74 – 5.69(m, 2H), 5.59 (dd, J = 12.6,3.0 Hz, 1H), 4.42 (dt, J = 11.2, 5.6 Hz, 4H), 4.22 (m, 4H), 4.06 (s, 3H),4.05 (s, 3H), 4.05 (s, 6H), 4.01 (s, 3H), 3.99 (s, 3H), 3.86 – 3.81 (m, 1H),3.76 – 3.70 (m, 1H), 3.14 (dd, J = 16.5, 12.7 Hz, 1H), 3.06 (dd, J = 16.5,11.6 Hz, 1H), 3.00 – 2.93 (m, 2H), 2.90 – 2.71 (m, 4H), 2.37 – 2.30 (m, 4H). 13C NMR (101 MHz, CDCl3) δ 187.58, 187.40, 174.75, 174.51, 174.37, 174.28,165.31, 165.19, 163.59, 163.58, 162.67, 162.46, 156.90, 156.64, 133.05,132.83, 130.36, 129.99, 129.66, 128.87, 125.93, 112.82, 112.70, 108.37,107.95, 106.14, 105.98, 89.42, 89.28, 78.88, 78.69, 66.58, 66.43, 60.40,56.99, 56.94, 56.77, 56.75, 56.55, 56.54, 52.67, 52.61, 52.32, 52.30, 44.53,43.39, 31.86, 31.67, 31.64, 31.54. HRMS (ESI) [M+H] + calcd for C 26 H 36 N2O9S2Na:663.0931, found: 663.0923.

[0109] Compound 9d: pale yellow solid, yield: 61.3%. 1 H NMR (400 MHz, CDCl3) δ 7.86 (dd, J =8.6, 2.2 Hz, 1H), 7.80 (d, J = 2.1 Hz, 1H), 7.13 (d, J = 8.7 Hz, 1H), 7.00(s, 1H), 6.95 (d, J = 6.2 Hz, 6H), 6.81 (d, J = 8.3 Hz, 2H), 6.72 (s, 1H), 6.08 (s, 1H), 5.28 (dd, J = 13.0, 3.4 Hz, 1H), 4.09 (s, 3H), 4.01 (s, 3H), 3.93 (s, 3H), 2.87 (dd, J = 16.7, 12.9 Hz, 1H), 2.78 (dd, J = 16.7, 3.4 Hz, 1H), 2.23 (s, 3H), 2.15 (s, 3H). 13C NMR (101 MHz, CDCl3) δ 187.61, 165.11,164.22, 162.61, 156.40, 135.73, 134.59, 134.35, 133.61, 133.14, 130.23,129.93, 129.83, 128.29, 126.62, 122.16, 120.14, 113.02, 108.31, 105.91,89.48, 78.99, 56.86, 56.72, 56.41, 44.57, 20.77, 20.75. HRMS (ESI) [M+H] + calcd for C 32 H 32 N2O9N a S2: 675.1447, found: 675.1444.

[0110] Compound 9e: pale yellow solid, yield: 72.8%. 1 H NMR (400 MHz, DMSO-d6) δ 10.28 (s,1H), 10.10 (s, 1H), 8.08 (d, J = 1.9 Hz, 1H), 7.74 (dd, J = 8.7, 1.9 Hz, 1H),7.29 – 7.02 (m, 9H), 6.31 (s, 1H), 5.70 (dd, J = 10.9, 3.4 Hz, 1H), 3.95 (s,3H), 3.88 (s, 3H), 3.88 (s, 3H), 2.99 (dd, J = 16.5, 11.0 Hz, 1H), 2.85 (dd,J = 16.4, 3.5 Hz, 1H). 13 C NMR (101 MHz, DMSO-d6) δ 187.48, 165.01, 163.52,163.04, 156.63, 138.05, 137.39, 133.38, 130.72, 129.34, 129.30, HRMS (ESI) [M+H] + calcd for C 30 H 27Cl2N2O9S2: 693.0535, found: 693.0540.

[0111] Compound 9f: pale yellow solid, yield: 56.5%. 1 H NMR (400 MHz, DMSO-d6) δ 10.31 (s,1H), 10.12 (s, 1H), 8.08 (d, J = 2.1 Hz, 1H), 7.74 (dd, J = 8.7, 2.1 Hz, 1H), 7.24 (dd, J = 9.5, 2.7 Hz, 2H), 7.21 – 7.16 (m, 2H), 7.16 – 7.07 (m, 3H), 6.32 (s, 1H), 5.70 (d, J = 11.0, 3.5 Hz, 1H), 3.94 (s, 3H), 3.88 (s, 3H), 3.87 (s, 3H), 3.00 (dd, J = 16.5, 11.1 Hz, 1H), 2.85 (dd, J = 16.4, 3.6 Hz,1H). 13 C NMR (101 MHz, DMSO-d6) δ 187.54, 165.02, 163.52, 163.38, 163.05,156.62, 138.05, 137.39, 133.42, 130.72, 129.38, 129.34, HRMS (ESI) [M+H] + calcd for C 30 H 24 Cl4N2O9S2: 760.9756, found:760.9761.

[0112] Compound 9g: pale yellow solid, yield: 53.8%. 1H NMR (400 MHz, CDCl3) δ 7.95 (d, J =2.2 Hz, 1H), 7.77 (dd, J = 8.7, 2.2 Hz, 1H), 7.33 – 7.27 (m, 4H), 7.20 (s,1H), 7.08 (dd, J = 8.9, 4.5 Hz, 2H), 7.01 – 6.96 (m, 2H), 6.93 – 6.85 (m,2H), 6.09 (s, 1H), 5.41 (dd, J = 11.2, 4.4 Hz, 1H), 4.05 (s, 3H), 4.02 (s,3H), 3.95 (s, 3H), 2.98 – 2.82 (m, 2H). 13 C NMR (101 MHz, CDCl3) δ 187.31,165.31, 163.98, 162.60, 156.38, 136.24, 135.61, 133.20, 132.41, 132.37,130.26, 128.57, HRMS (ESI) [M+H] + calcd for C 30 H 27 Br2N2O9S2: 782.9504, 780.9525, found: 782.9500, 780.9521.

[0113] Compound 9h: pale yellow solid, yield: 54.9%. 1H NMR (400 MHz, CDCl3) δ 7.94 (d, J =2.1 Hz, 1H), 7.86 (dd, J = 8.7, 2.1 Hz, 1H), 7.32 (s, 1H), 7.24 (t, J = 1.8Hz, 1H), 7.23 (t, J = 1.9 Hz, 1H), 7.21 (s, 1H), 7.17 – 7.10 (m, 3H), 7.09 –7.00 (m, 3H), 6.89 (dd, J = 8.1, 1.2 Hz, 1H), 6.10 (s, 1H), 5.41 (dd, J =11.7, 4.3 Hz, 1H), 4.05 (s, 3H), 4.04 (s, 3H), 3.95 (s, 3H), 2.97 – 2.80 (m,2H). 13 C NMR (101 MHz, CDCl3) δ 187.46, 165.49, 164.13, 162.91, 156.44,138.71, 138.00, 133.25, 130.70, 130.68, 130.36, 128.36, 128.27, 127.38,126.30, 123.87, 122.82, 122.70, 121.88, 119.33, 117.75, 113.14, 107.74,105.99, 89.49, 78.91, 56.97, 56.71, 56.50, 44.64. HRMS (ESI) [M+H] + , calcdfor C 30 H 27 Br2N2O9S2: 782.9504, 780.9525, found: 782.9505, 780.9522.

[0114] Compound 9i: pale yellow solid, yield: 54.6%. 1H NMR (400 MHz, DMSO-d6) δ 10.06 (s,1H), 9.87 (s, 1H), 7.99 (d, J = 2.1 Hz, 1H), 7.74 (dd, J = 8.7, 2.1 Hz, 1H), 7.21 (d, J = 8.8 Hz, 1H), 7.15 – 7.09 (m, 4H), 7.04 – 6.97 (m, 4H), 6.31 (s,1H), 5.64 (dd, J = 11.0, 3.5 Hz, 1H), 3.95 (s, 3H), 3.89 (s, 3H), 3.87 (s,3H), 2.94 (dd, J = 16.5, 11.1 Hz, 1H), 2.81 (dd, J = 16.5, 3.7 Hz, 1H). 13 CNMR (101 MHz, DMSO-d6) δ 187.51, 164.94, 163.53, 162.98, 158.95 (d, J C,F =238.4 Hz), 158.82 (d, J C,F = 238.4 Hz), 156.58, 134.92 (d, J C,F = 64.9 Hz), 134.90 (d, J C,F = 65.0 Hz), 133.25, 130.73, 129.05, 126.58, 121.72 (d, J C,F =8.2 Hz), 121.05 (d, J C,F = 8.0 Hz), 116.18, 115.96, 113.37, 108.14, 105.73,90.51, 78.06, 57.44, 56.85, 56.79, 44.20. HRMS (ESI) [M+H] + , calcd forC 30 H 26 F2N2O9S2Na: 683.0945, found: 683.0947.

[0115] Compound 9j: pale yellow solid, yield: 55.8%. 1H NMR (400 MHz, DMSO-d6) δ 10.06 (s,1H), 9.85 (s, 1H), 8.05 (d, J = 2.2 Hz, 1H), 7.76 (dd, J = 8.7, 2.1 Hz, 1H), 7.20 (d, J = 8.8 Hz, 1H), 7.19 – 7.09 (m, 8H), 6.97 – 6.89 (m, 2H), 6.31 (s,1H), 5.63 (dd, J = 11.3, 3.5 Hz, 1H), 3.95 (s, 3H), 3.89 (s, 3H), 3.86 (s,3H), 2.92 (dd, J = 16.5, 11.3 Hz, 1H), 2.80 (dd, J = 16.5, 3.6 Hz, 1H). 13 CNMR (101 MHz, DMSO-d6) δ 187.52, 164.91, 163.59, 163.09, 156.62, 139.02,138.37, 133.14, 130.77, 129.40, 128.99, 126.88, 123.69, 123.50, 119.34,118.83, 113.43, 108.38, 105.74, 90.56, 78.15, 57.43, 56.83, 56.79, 44.41.HRMS (ESI) [M+H] + calcd for C 30 H 28 N2O9S2Na: 647.1134, found: 647.1129.

[0116] Compound 9k: pale yellow solid, yield: 50.6%. 1H NMR (400 MHz, CDCl3) δ 7.91 – 7.82(m, 2H), 7.11 (d, J = 8.8 Hz, 1H), 7.08 (d, J = 8.3 Hz, 2H), 7.04 (d, J = 7.9Hz, 1H), 6.89 (s, 1H), 6.69 (t, J = 2.1 Hz, 1H), 6.64 (dd, J = 8.0, 1.4 Hz,1H), 6.61 (t, J = 2.1 Hz, 1H), 6.60 – 6.50 (m, 2H), 6.45 (dd, J = 7.9, 1.5Hz, 1H), 6.09 (s, 1H), 5.34 (dd, J = 12.3, 3.8 Hz, 1H), 4.07 (s, 3H), 4.02(s, 3H), 3.94 (s, 3H), 3.69 (s, 3H), 3.64 (s, 3H), 2.88 (dd, J = 16.7, 12.4Hz, 1H), 2.81 (dd, J = 16.7, 3.9 Hz, 1H). 13 C NMR (101 MHz, CDCl3) δ 187.54,165.24, 164.19, 162.74, 160.28, 160.23, 156.40, 138.39, 137.62, 133.13,130.30, 130.07, 130.04, 128.23, 126.53, 113.38, 113.05, 111.65, 110.80,110.02, 108.21, 107.58, 105.97, 105.25, 89.48, 78.96, 56.87, 56.68, 56.43,56.43, 55.28, 55.20, 44.66. HRMS (ESI) [M+H] + calcd for C 32 H 32 N2O 11 S2Na:707.1345, found: 707.1345.

[0117] Compound 9l: pale yellow solid, yield: 51.3%. 1H NMR (400 MHz, DMSO-d6) δ 9.69 (s,1H), 9.47 (s, 1H), 7.90 (d, J = 2.1 Hz, 1H), 7.71 (dd, J = 8.7, 2.1 Hz, 1H), 7.20 (d, J = 8.8 Hz, 1H), 7.04 (d, J = 9.1 Hz, 2H), 7.01 (d, J = 9.1 Hz, 2H), 6.75 (d, J = 6.0 Hz, 2H), 6.73 (d, J = 6.0 Hz, 2H), 6.31 (s, 1H), 5.56 (dd, J= 10.8, 3.8 Hz, 1H), 3.97 (s, 3H), 3.92 (s, 3H), 3.86 (s, 3H), 3.64 (s, 3H), 3.61 (s, 3H), 2.87 (dd, J = 16.5, 10.9 Hz, 1H), 2.79 (dd, J = 16.5, 3.9 Hz, 1H). 13 C NMR (101 MHz, DMSO-d6) δ 187.54, 164.79, 163.56, 162.92, 156.51,156.39,156.20, 132.98, 131.53, 130.88, 130.77, 128.81, 126.91, 122.48,121.79, 114.62 114.58, 113.24, 108.36, 105.76, 90.46, 78.00, 57.39, 56.81,56.72, 55.53, 55.48, 44.23. HRMS (ESI) [M+H] + calcd for C 32 H 32 N2O 11 S2Na:707.1345, found: 707.1338.

[0118] Compound 9m: pale yellow solid, yield: 51.6%. 1H NMR (400 MHz, CDCl3) δ 7.94 (d, J =2.0 Hz, 1H), 7.85 (dd, J = 8.6, 2.0 Hz, 1H), 7.13 (d, J = 8.7 Hz, 1H), 6.87(s, 1H), 6.76 (s, 1H), 6.32 (s, 2H), 6.28 (s, 2H), 6.10 (s, 1H), 5.45 (t, J =7.9 Hz, 1H), 4.08 (s, 3H), 4.01 (s, 3H), 3.96 (s, 3H), 3.74 (s, 9H), 3.72 (s,3H), 3.68 (s, 6H), 2.90 (s, 1H), 2.88 (d, J = 1.6 Hz, 1H). 13 C NMR (101 MHz, CDCl3) δ 187.15, 165.28, 164.08, 162.64, 156.34, 153.55, 153.48, 136.13,135.24, 133.12, 133.02, 132.04, 130.52, 127.89, 126.96, 113.03, 108.31,106.14, 99.93, 97.62, 89.53, 78.69, 60.90, 60.83, 56.83, 56.77, 56.47, 56.19,56.09, 44.44. HRMS (ESI) [M+H] + calcd for C 36 H 40 N2O 15 S2Na: 827.1768, found:827.1758.

[0119] Compound 9n: pale yellow solid, yield: 55.1%. 1H NMR (400 MHz, CDCl3) δ 7.89 (d, J =2.2 Hz, 1H), 7.81 (dd, J = 8.6, 2.2 Hz, 1H), 7.66 – 7.55 (m, 2H), 7.17 (d, J= 2.6 Hz, 1H), 7.11 (d, J = 8.6 Hz, 2H), 6.84 – 6.66 (m, 4H), 6.09 (s, 1H),5.40 (dd, J = 12.0, 3.9 Hz, 1H), 4.04 (s, 3H), 4.00 (s, 3H), 3.96 (s, 3H),2.91 – 2.78 (m, 2H). 13 C NMR (101 MHz, CDCl3) δ 187.37, 165.35, 163.89,162.73, 159.71 (dd, J C,F = 242.2, 11.2 Hz), 159.66 (dd, J C,F = 246.6, 11.3 Hz),156.53, 153.95 (dd, J C,F = 245.4, 12.0 Hz), 153.70 (dd, J C,F = 244.5, 12.1 Hz),133.10, 132.79, 129.98, 128.67, 128.38, 126.31, 124.89 (dd, J C,F = 9.5, 1.5Hz), 124.78 (dd, J C,F = 9.4, 1.9 Hz), 121.19 (dd, J C,F = 12.3, 3.8 Hz), 120.78(dd, J C,F = 12.4, 3.8 Hz), 112.68, 111.94 (dd, J C,F = 22.4, 3.8 Hz), 108.04,105.82, 103.92 (t, J C,F = 23.6 Hz), 103.90 (t, J C,F = 23.7 Hz), 103.66, 89.16,78.66, 56.79, 56.51, 56.47, 44.35. HRMS (ESI) [M+H] + , calcd for C30 H 25 F4N2O9S2:697.0938, found: 697.0932.

[0120] Compound 9o: pale yellow solid, yield: 43.9%. 1 H NMR (400 MHz, CDCl3) δ 7.91 (dd, J =8.6, 2.0 Hz, 1H), 7.81 (d, J = 2.0 Hz, 1H), 7.23 (d, J = 7.7 Hz, 1H), 7.20 (d, J = 7.9 Hz, 1H), 7.12 (d, J = 8.7 Hz, 1H), 7.09 (d, J = 7.6 Hz, 1H), 7.05– 7.02 (m, 2H), 7.00 – 6.91 (m, 3H), 6.82 (s, 1H), 6.68 (s, 1H), 6.12 (s,1H), 5.32 (dd, J = 12.2, 3.8 Hz, 1H), 4.04 (s, 3H), 3.99 (s, 3H), 3.96 (s, 3H), 2.95 – 2.77 (m, 2H), 2.26 (s, 3H), 1.97 (s, 3H). 13 C NMR (101 MHz, CDCl3)δ 187.56, 165.18, 164.06, 162.67, 156.38, 135.42, 134.71, 133.21, 130.92,130.88, 130.58, 130.25, 128.88, 128.00, 127.53, 127.01, 126.90, 125.75,124.90, 122.33, 120.37, 113.03, 109.31, 106.05, 89.55, 78.80, 56.82, 56.58,56.46, 44.48, 17.60, 17.32. HRMS (ESI) [M+H] + calcd for C 32 H 32 N2O9N a S2:675.1447, found: 675.1451.

[0121] Compound 9p: pale yellow solid, yield: 43.3%. 1H NMR (400 MHz, CDCl3) δ 7.90 – 7.84(m, 2H), 7.12 (d, J = 8.1 Hz, 2H), 7.03 (m, 2H), 6.92 (s, 1H), 6.90 – 6.80(m, 4H), 6.78 (s, 1H), 6.74 (d, J = 8.1 Hz, 1H), 6.09 (s, 1H), 5.31 (dd, J =12.2, 3.8 Hz, 1H), 4.08 (s, 3H), 4.01 (s, 3H), 3.93 (s, 3H), 2.96 – 2.75 (m,2H), 2.21 (s, 3H), 2.18 (s, 3H). 13 C NMR (101 MHz, CDCl3) δ 187.60, 165.18,164.23, 162.71, 156.42, 139.34, 139.21, 137.07, 136.32, 133.06, 130.30,129.09, 128.13, 126.69, 126.46, 125.47, 122.33, 120.18, 118.37, 116.57,113.05, 108.31, 105.93, 89.48, 78.95, 56.86, 56.68, 56.43, 44.69, 21.38,21.34. HRMS (ESI) [M+H] + calcd for C 32 H 32 N2O9N a S2: 675.1447, found: 675.1450.

[0122] Compound 9q: pale yellow solid, yield: 42.8%. 1H NMR (400 MHz, CDCl3) δ 7.99 (d, J =2.2 Hz, 1H), 7.85 (dd, J = 8.7, 2.2 Hz, 1H), 7.71 – 7.67 (m, 2H), 7.58 (s,1H), 7.46 (s, 1H), 7.25 – 7.09 (m, 4H), 7.07 (d, J = 8.7 Hz, 1H), 6.94 (t, J= 7.7 Hz, 2H), 6.08 (s, 1H), 5.39 (dd, J = 11.9, 4.2 Hz, 1H), 4.01 (s, 6H), 3.95 (s, 3H), 2.93 – 2.78 (m, 2H). 13 C NMR (101 MHz, CDCl3) δ 187.48, 165.43,164.13, 163.10, 156.55, 134.27, 133.75, 133.11, 130.07, 129.44, 129.41,128.51, 128.20, 127.97, 126.37, 125.07, 124.73, 123.26, 122.65, 120.45,119.53, 112.68, 108.07, 105.87, 89.19, 78.75, 56.80, 56.46, 56.41, 44.71.HRMS (ESI) [M+H] + calcd for C 30 H 26 Cl2N2O9S2Na: 715.0354, found: 715.0352.

[0123] Compound 9r: pale yellow solid, yield: 40.9%. 1 H NMR (400 MHz, CDCl3) δ 7.94 (d, J =2.2 Hz, 1H), 7.85 (dd, J = 8.7, 2.2 Hz, 1H), 7.30 (s, 1H), 7.17 (s, 1H), 7.15– 6.97 (m, 8H), 6.84 (dd, J = 8.0, 1.5 Hz, 1H), 6.10 (s, 1H), 5.40 (dd, J =11.8, 4.2 Hz, 1H), 4.06 (s, 3H), 4.03 (s, 3H), 3.95 (s, 3H), 2.97 – 2.80 (m,2H).13 C NMR (101 MHz, CDCl3) δ 187.44, 165.45, 164.13, 162.85, 156.44,138.55, 137.84, 134.85, 134.80, 133.25, 130.42, 130.39, 130.35, 128.35,126.36, 125.40, 124.52, 121.07, 119.14, 118.90, 117.34, 113.14, 107.85,106.00, 89.50, 78.92, 56.95, 56.72, 56.49, 44.60. HRMS (ESI) [M+H] + , calcdfor C 30 H 26 Cl2N2O9S2Na: 715.0354, found: 715.0349.

[0124] Compound 9S: pale yellow solid, yield: 42.8%. 1 H NMR (400 MHz, CDCl3) δ 7.90 (d, J =2.2 Hz, 1H), 7.85 (dd, J = 8.6, 2.3 Hz, 1H), 7.67 – 7.55 (m, 2H), 7.30 (d, J= 3.2 Hz, 1H), 7.18 (d, J = 3.3 Hz, 1H), 7.09 (d, J = 8.7 Hz, 1H), 7.04 –6.92 (m, 6H), 6.07 (s, 1H), 5.35 (dd, J = 12.7, 3.5 Hz, 1H), 4.04 (s, 3H), 3.99 (s, 3H), 3.94 (s, 3H), 2.88 (dd, J = 16.6, 12.8 Hz, 1H), 2.80 (dd, J =16.6, 3.6 Hz, 1H). 13 C NMR (101 MHz, CDCl3) δ 187.53, 165.31, 163.98, 162.89,156.56, 153.55 (d, J C,F = 242.2 Hz), 152.24 (d, J C,F = 242.2 Hz), 133.10, 130.03,128.39, 126.35, 125.66 (d, J C,F= 19.7 Hz), 125.58 (d, J C,F = 19.7 Hz), 125.17(d, J C,F = 11.8 Hz), 124.91 (d, J C,F =3.5 Hz), 124.87 (d, J C,F =3.5 Hz), 124.70(d, J C,F = 12.0 Hz), 122.69, 122.38, 115.37 (dd, J C,F = 2.9 Hz), 115.18 (dd,J C,F = 2.8 Hz), 112.75, 108.15, 105.82, 89.21, 78.84, 56.78, 56.49, 56.45,44.64. HRMS (ESI) [M+H] + calcd for C 30 H 26 F2N2O9S2Na: 683.0945, found: 683.0945.

[0125] Compound 9t: pale yellow solid, yield: 46.8%. 1 H NMR (400 MHz, DMSO-d6) δ 10.38 (s,1H), 10.22 (s, 1H), 8.11 (d, J = 2.0 Hz, 1H), 7.79 (dd, J = 8.7, 2.0 Hz, 1H),7.25 – 7.13 (m, 3H), 6.96 – 6.88 (m, 4H), 6.79 – 6.73 (m, 2H), 6.33 (s, 1H), 5.68 (dd, J = 11.4, 3.4 Hz, 1H), 3.95 (s, 3H), 3.89 (s, 6H), 2.95 (dd, J =16.5, 11.5 Hz, 1H), 2.83 (dd, J = 16.5, 3.5 Hz, 1H). 13 C NMR (101 MHz, DMSO-d6) δ 187.48, 165.12, 163.59, 163.20, 162.73 (d, J C,F = 240.9 Hz), 162.65 (d,J C,F= 241.3 Hz), 156.65, 140.99 (d, J C,F = 10.7 Hz), 140.27 (d, J C,F = 10.6Hz), 133.34, 131.18, 131.08, 130.85, 128.96, 126.53, 114.71 (d, J C,F = 2.6Hz), 114.19 (d, J C,F = 2.1 Hz), 113.57, 110.17 (d, J C,F = 20.9 Hz), 109.79 (d,J C,F = 20.9 Hz), 108.00, 105.85 (d, J C,F = 23.9 Hz), 105.74, 105.08 (d, J C,F =25.8 Hz), 90.70, 78.27, 57.50, 56.89, 56.84, 44.51. HRMS (ESI) [M+H] + , calcdfor C 30 H 26 F2N2O9S2Na: 683.0945, found: 683.0939.

[0126] Example 2: Cell proliferation experiment

[0127] Gastric cancer cells HGC27, esophageal squamous cell carcinoma cells KYSE70 and KYSE450, colorectal cancer cells HCT116, and lung cancer cells A549 were seeded in 96-well plates, approximately 4000 cells per well, and incubated overnight at 37°C with 5% CO2. Then, different concentrations of 2'-hydroxy-4,4',6'-trimethoxychalcone and its derivatives (0.1–40 μM) were added for drug screening. The efficacy of 2'-hydroxy-4,4',6'-trimethoxychalcone and its derivatives (4b, 5, 7e, 7k, 11e, 11g) was compared by adding different concentrations of 2'-hydroxy-4,4',6'-trimethoxychalcone and its derivatives (4b, 5, 7e, 7k, 11e, 11g) at 1–8 μM. After drug treatment for 0, 24, 48, and 72 h, the cells were incubated with MTT for 2 h, the supernatant was discarded, 150 μL of DMSO was added, and the absorbance was measured at 490 nm. The IC50 of each derivative was calculated. 50 Values ​​and results are detailed in the attached document. Figure 9 See Table 1.

[0128] Table 1. Inhibition of esophageal cancer, colon cancer, and gastric cancer cell lines by the derivatives of this invention. 50 (μM) In vitro proliferation activity

[0129] Example 3: Plate Colony Formation Experiment

[0130] Gastric cancer cells HGC27, esophageal squamous cell carcinoma cells KYSE450, and colorectal cancer cells HCT116 (900 cells per well) were seeded in 6-well plates, and compound 3b and its derivatives 6i, 9e, 9i, and 9l were added at concentrations of 1, 2, 4, and 8 μM. The plates were then incubated at 37 ℃ and 5% CO2 for 10 days. The number of cell clones was photographed and counted, and the statistical results were analyzed.

[0131] Example 4: In vivo antitumor activity experiment in mice

[0132] HGC27 cells were digested and centrifuged, counted, and then centrifuged again. Based on the counting results, an appropriate amount of PBS was added to adjust the cell concentration to 5 × 10⁻⁶ cells / year. 7 / ml, place the cell suspension on ice, and after entering the animal room, inoculate each nude mouse with 100 μL of cell suspension in a clean bench, which is approximately 5 × 10⁶ cells per nude mouse. 6 A mouse model of human gastric cancer HGC27 cell xenograft tumor (CDX) was established using 10 cells.

[0133] One week after inoculation, nude mice were ear-tagged and weighed. Mice were then evenly distributed into groups of seven mice each, and the nine groups were administered physiological saline containing 10% DMSO, compound 6i, 9e, 9i, and 9l at doses of 2.5, 5 mg / kg / day, respectively, via gavage. Medication was administered daily, and tumor appearance was monitored. Once tumors became visible to the naked eye, tumor size was measured. Tumor volume and body weight were measured at fixed intervals. The control group's tumor volume was reduced to 1000 mm². 3 At that point, the experiment was terminated, the tumor tissue was removed, the tumor was weighed, and a photograph was taken.

[0134] Example 5: Hydrogen bromide-activated Sepharose 4B protein pull-down assay

[0135] A drug bead mixture was prepared by combining 2 mg of compounds 3b, 6i, 9e, 9i, 9l and 0.3 g of Sepharose 4B beads. The bead mixture was then mixed with HGC27 gastric cancer cell lysate and incubated overnight at 4°C. After washing, the mixture was incubated in a 95°C metal bath for 5 min. Simultaneously, a 10% input sample was prepared. The sample was then subjected to SDS-PAGE. After gel running, Coomassie Brilliant Blue staining was performed and the stain was destained. Differential bands were excised and sent to Jingjie Biotechnology Co., Ltd. for protein proteomic analysis. The obtained data were analyzed using Venn and Metascape to form a candidate protein interaction network. The sample prepared with HGC27 gastric cancer cell lysate and the bead mixture was then subjected to Western blotting detection and incubated with YBX1 and RPN1 antibodies.

[0136] Experimental results

[0137] Table 1 shows the growth inhibition of the derivatives of this invention on esophageal squamous cell carcinoma cells, gastric cancer cells, colorectal cancer cells, and lung cancer cells. The results indicate that most of the modified derivatives showed superior activity compared to 2'-hydroxy-4,4',6'-trimethoxychalcone, with the inhibitory activity exhibiting a clear time- and concentration-dependent effect. Compared to derivatives of general formula I (6a ~ 6m), most derivatives of general formula II (9a ~ 9t) were superior. Specifically, the IC50 of derivative 6i in general formula I against the above-mentioned cancer cells was [not specified in the original text]. 50 Within the range of 4.80–13.18 μM, the IC50 values ​​of derivatives 9d, 9e, 9f, 9g, 9i, 9j, 9l, 9o, 9p, and 9t of general formula II against the above-mentioned cancer cells were [not specified in the original text]. 50 Derivative 9j exhibited the best relative activity in the ranges of 2.08 ~ 3.28, 2.25 ~ 3.23, 1.92 ~ 4.25, 2.53 ~ 4.86, 1.49 ~ 4.49, 0.90 ~ 3.94, 2.25 ~ 6.60, 2.39 ~ 3.92, 2.39 ~ 4.79, and 2.64 ~ 5.22 μM, respectively.

[0138] Figure 9 The growth inhibition of esophageal squamous cell carcinoma cells, gastric cancer cells, and colorectal cancer cells by derivatives 6i, 9e, 9i, 9l, and 3b of the present invention is shown. The modified derivatives have better antitumor activity than 2'-hydroxy-4,4',6'-trimethoxychalcone. Figure 9The bar graphs show tumor cell proliferation at different time points with different drug concentrations. The results indicate that the derivatives 6i, 9e, 9i, and 9l of this invention, at concentrations ranging from 1 to 8 μM, can inhibit the proliferation of esophageal squamous cell carcinoma KYSE450, gastric cancer cells HGC27, and colorectal cancer cells HCT116, with the highest inhibition rate at a concentration of 8 μM. (Compared with the control group, * indicates P < 0.05, ** indicates P < 0.01, and *** indicates P < 0.001)

[0139] Figure 10 The invention describes the inhibitory effects of derivatives 6i, 9e, 9i, 9l, and 3b on the colony formation of esophageal squamous cell carcinoma cells, gastric cancer cells, and colorectal cancer cells. The modified derivatives showed better colony formation inhibition than 2'-hydroxy-4,4',6'-trimethoxychalcone. Figure 10 This document presents statistics on the number of clones and images showing the clone formation of esophageal squamous cell carcinoma KYSE450, gastric cancer cells HGC27, and colorectal cancer cells HCT116 inhibited by derivatives 6i, 9e, 9i, and 9l of this invention. The results indicate that each compound inhibited the clone formation of esophageal cancer cells KYSE450, gastric cancer cells HGC27, and colorectal cancer cells HCT116 to varying degrees, exhibiting a concentration-dependent effect. Derivative 9i showed the best activity. (Compared to the control group, * indicates P<0.05, ** indicates P<0.01, *** indicates P<0.001)

[0140] Figure 11 This paper presents the therapeutic effects of derivatives 6i, 9e, 9i, and 9l of the present invention on tumor growth in a mouse model of human gastric cancer HGC27 cell xenografts. A shows the trend of tumor volume changes during drug administration; B shows the tumor weight after mouse sacrifice at the end of the experiment; C shows tumor images of the control and drug-treated groups. The results indicate that, based on tumor volume, weight, and number analysis, compounds 6i, 9e, 9i, and 9l at 2.5 and 5 mg / kg / day significantly inhibited the growth of tumors in the small model of human gastric cancer HGC27 cell xenografts, demonstrating significant therapeutic effects. D shows the trend of mouse body weight changes during drug administration; E, F, and G show the weights of the liver, spleen, and kidneys of mice in each group after the experiment. The results indicate that 2.5 and 5 mg / kg / day of these compounds significantly inhibited tumor growth in the mouse model of human gastric cancer HGC27 cell xenografts. Compounds 6i, 9e, 9i, and 9l (mg / kg / day) had no effect on body weight, liver, spleen, or kidney in mice, indicating that derivatives 6i, 9e, 9i, and 9l had better therapeutic effects and no obvious toxic side effects compared with the control group (* indicates P<0.05, ** indicates P<0.01, and *** indicates P<0.001 compared with the control group).

[0141] Figure 12The derivatives 9e, 9i, 9l, 6i, and 3b of this invention can bind well to the target proteins YBX1 and RPN1. A is the Venn diagram of mass spectrometry results, and B is the Metascape protein interaction network diagram of mass spectrometry results, indicating that the derivatives 9e, 9i, 9l, 6i, and 3b of this invention may target YBX1 and RPN1. C and D are the hydrogen bromide-activated Sepharose 4B protein pull-down experiments in human gastric cancer HGC27 cells, indicating that compound 3b and derivatives 6i, 9e, 9i, and 9l can bind well to the target proteins YBX1 and RPN1.

[0142] In summary, the MTT assay was used to test cell proliferation at 0, 24, 48, and 72 h after treatment with the derivatives of formulas I and II. The results confirmed that the derivatives inhibited cell proliferation in esophageal cancer cell lines, gastric cancer cell lines, and colorectal cancer cell lines, with activity superior to 2'-hydroxy-4,4',6'-trimethoxychalcone. Plate colony formation assays showed that compounds 6i, 9e, 9i, and 9l inhibited the formation of esophageal KYSE450, gastric cancer HGC27, and colorectal cancer HCT116 cell colonies. Hydrogen bromide-activated Sepharose 4B protein pull-down assays and proteomic analysis showed that derivatives 6i, 9e, 9i, and 9l could bind to the target proteins YBX1 and RPN1. In the HGC27 cell CDX model of gastric cancer, derivatives 6i, 9e, 9i, and 9l showed significant tumor therapeutic effects on mice with gastric cancer cell xenografts, providing assistance for clinical research on drugs for the prevention and treatment of esophageal cancer, gastric cancer, colorectal cancer, and other tumors.

Claims

1. 2'-hydroxy-4,4',6'-trimethoxychalcone and 5,7,4'-trimethoxyflavanone sulfonamide derivatives, characterized in that, It has the structure shown in the following general formula: R1, R2, and R3 are each independently selected from ethyl, cyclopropyl, 2,4-difluorophenyl, 4-fluorophenyl, 4-chlorophenyl, 2,3-dichlorophenyl, 3-bromophenyl, phenyl, 4-methoxyphenyl, 4-methylphenyl, 4-bromophenyl, 4-nitrophenyl, 4-cyanophenyl, 3-bromophenyl, 3-methoxyphenyl, homoserine lactone, 3-chlorophenyl, 2-fluorophenyl, 3,4,5-trimethoxyphenyl, 2-chlorophenyl, 2-methylphenyl, 3-methylphenyl, 3-chlorophenyl, 2-fluorophenyl, and 3-fluorophenyl. 2,2'-hydroxy-4,4',6'-trimethoxychalcone and 5,7,4'-trimethoxyflavanone sulfonamide derivatives, characterized in that, Specifically, compounds with the following structures:

3. The method for preparing the 2'-hydroxy-4,4',6'-trimethoxychalcone and 5,7,4'-trimethoxyflavanone sulfonamide derivatives (wherein R1, R2, and R3 are defined as in claim 1) according to claim 1, characterized in that, The synthesis route is as follows: The reaction route for general formula I is as follows: The synthesis process of general formula I is as follows: (1) Dissolve 2-hydroxy-4,6-dimethoxyacetophenone (compound 1) and 4-methoxy-3-nitrobenzaldehyde (compound 2a) in the reaction solvent, add alkali, heat to reflux, and a Claisen-Schmidt reaction occurs. After the reaction is complete, cool to room temperature, pour the reaction solution into ice water, adjust the pH of the system to 5-6 with dilute hydrochloric acid, precipitate out, filter, wash, and obtain 3-nitro-2'-hydroxy-4,4',6'-trimethoxychalcone (compound 3a), which can be directly used in the next step of the reaction; (2) Compound 3a was dissolved in the reaction solvent, and reducing metal powder was added. Acid was added and refluxed to reduce the nitro group to an amino group. The mixture was purified by silica gel column chromatography to obtain 3-amino-2'-hydroxy-4,4',6'-trimethoxychalcone (compound 4). (3) Compound 4 was dissolved in the reaction solvent, and an acylation reaction catalyst and a base were added. The mixture was then reacted with commercially available sulfonyl chloride (compound 5) to synthesize a series of derivatives 6 of general formula I. The reaction route for formula II is as follows: The synthesis process of Formula II is as follows: (1) Dissolve 2-hydroxy-4,6-dimethoxyacetophenone (compound 1) and p-anisaldehyde (compound 2b) in a reaction solvent, add alkali, heat to 110 ± 10℃ to carry out the Claisen-Schmidt reaction. After the reaction is complete, cool to room temperature, pour the reaction solution into ice water, adjust the pH of the system to 5 ~ 6 with dilute hydrochloric acid, precipitate, filter, wash, recrystallize to obtain 2'-hydroxy-4,4',6'-trimethoxychalcone (compound 3b); (2) At 0 ± 5℃, compound 3b was added to the sulfonating agent. After the addition was complete, the mixture was moved to room temperature to sulfonate compound 3b. After the reaction was complete, the reaction was quenched by ice water, and a precipitate was formed. The precipitate was filtered, washed with pure water, and a solid was obtained. The prepared sulfonyl chloride (compound 7) can be used directly in the next step of the reaction. (3) Dissolve compound 7 in the reaction solvent, add commercially available amine compound 8, stir at room temperature until the reaction is complete, evaporate to dryness, dissolve the residue in ethyl acetate, wash with hydrochloric acid solution until neutral, wash with saturated brine, dry with anhydrous sodium sulfate, filter, concentrate, and purify by silica gel column chromatography to obtain a series of derivatives 9 of general formula II.

4. The method for preparing the derivative of general formula I according to claim 3, characterized in that, In step (1), the molar ratio of Claisen-Schmidt reaction compound 1, compound 2a, and base is 1:(1.2~1.4):(2~5), and the solvent used is methanol, ethanol, tert-butanol, or acetonitrile; the base used is one or a mixture of two or more of potassium carbonate, sodium carbonate, sodium hydroxide, lithium hydroxide, and sodium hydride; the concentration of dilute hydrochloric acid is 5~ 10 mol / L; the recrystallization solvent is methanol, ethanol or tert-butanol; in step (2), the molar ratio of compound 4a, reducing metal powder and acid is 1:(5~8):(5~10), the selected reaction solvent is methanol, ethanol, isopropanol or tert-butanol, the reducing metal powder is iron powder, magnesium powder or zinc powder, and the acid is concentrated hydrochloric acid, concentrated sulfuric acid or glacial acetic acid; in step (3), the molar ratio of compound 4, commercially available sulfonyl chloride (compound 5), catalyst and base is 1:(1.5~2):(1~2):(3~5), the selected reaction solvent is acetone, tetrahydrofuran or 1,4-dioxane, the catalyst is DMAP or DCC, and the base is triethylamine, pyridine or DBU.

5. The method for preparing the general formula II derivative according to claim 3, characterized in that, In step (1), the molar ratio of Claisen-Schmidt reaction compound 1, compound 2b, and base is 1:(1.2~1.4):(2~5), and the solvent used is methanol, ethanol, tert-butanol, or acetonitrile; the base used is one or a mixture of two or more of potassium carbonate, sodium carbonate, sodium hydroxide, lithium hydroxide, and sodium hydride; the concentration of dilute hydrochloric acid is 5~10 mol / L; the recrystallization solvent is methanol, ethanol, or tert-butanol; in step (2), the molar ratio of compound 1 and chlorosulfonic acid is 1:(17~18); in step (3), the molar ratio of compound 7 and commercially available amine (compound 8) is 1:(1.5~2), and the solvent used is acetone, tetrahydrofuran, or 1,4-dioxane.

6. The use of 2'-hydroxy-4,4',6'-trimethoxychalcone sulfonamide and 5,7,4'-trimethoxyflavanone derivatives and their salts as described in claim 1 or 2 in the preparation of medicaments for treating gastric cancer, esophageal cancer, colorectal cancer or lung cancer.

7. The application according to claim 6, characterized in that, The antitumor drugs mentioned are drugs for treating esophageal cancer, colorectal cancer, gastric cancer, and lung cancer.

8. The application according to claim 6, characterized in that, It can bind to the target protein YBX1 / RPN1.

9. The application according to claim 8, characterized in that, It can be used as an active ingredient or in combination with other drugs to prepare single or compound formulations targeting the YBX1 / RPN1 protein, following conventional pharmaceutical methods and processes.

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