Anti-tumor compound and derivative thereof as well as preparation method and application of anti-tumor compound and derivative thereof

Abstract

The invention relates to an anti-tumor compound and a derivative or pharmaceutically acceptable salt and stereoisomer thereof, and a pharmaceutical composition containing the anti-tumor compound and the derivative or pharmaceutically acceptable salt and stereoisomer thereof. According to the present invention, the parent nucleus of the compound is diphenylethyl 3, 3 '-(4, 5-dihydroxy-1, 2-phenylene) (2E, 2' E)-diacrylate by using the IUPAC standard naming method, and it has been proved that the compound and the derivative thereof have good anti-tumor effects, and can effectively inhibit the proliferation of a variety of tumor cells. The invention also relates to a preparation method of the compound and application of the compound in preparation of medicines for preventing and / or treating tumors.

Description

Technical Field

[0001] This invention belongs to the field of pharmaceutical technology, specifically relating to an antitumor compound and its derivatives, as well as their preparation methods and uses. Background Technology

[0002] A tumor is a new growth formed by the abnormal proliferation of local tissue cells under the influence of tumorigenic factors. Based on biological behavior, tumors can be divided into benign and malignant tumors. Benign tumors typically grow slowly, have well-defined borders, and do not metastasize (such as lipomas), while malignant tumors (i.e., cancer) are characterized by invasive growth, indistinct borders, and metastasis. They can originate from epithelial tissue (such as lung cancer) or mesenchymal tissue, posing a greater threat to the body and often accompanied by systemic symptoms such as weight loss and fatigue. As the main type of malignant tumor, cancer requires comprehensive treatment including surgery, radiotherapy, chemotherapy, targeted therapy, and immunotherapy. Its prognosis is worse than that of benign tumors, and it is prone to recurrence and metastasis.

[0003] Significant progress has been made in global anti-tumor drug development in recent years. In 2024, China approved 48 Class 1 innovative drugs, with anti-tumor drugs accounting for the largest share (42.54%), covering immunotherapy (such as PD-1 / PD-L1 inhibitors), antibody-drug conjugates (ADCs), bispecific antibodies, and "cancer-agnostic" therapies (such as targeted drugs against NTRK / RET gene fusions). The rise of precision medicine has driven cancer treatment to shift from organ-based approaches to molecular subtyping, such as personalized treatment plans based on tumor mutational burden (TMB) or specific gene mutations.

[0004] The development of novel anti-tumor drugs is crucial for improving patient survival. While survival rates have improved for some cancer types, most advanced cancers still lack effective treatments, and drug resistance and tumor heterogeneity pose significant challenges. The application of artificial intelligence and multi-omics technologies is accelerating drug development, while the expanding global demand for medical services (especially in emerging markets like China) further underscores the urgency of innovative therapies. Through policy support and technological breakthroughs, novel drugs are expected to provide more patients with accessible and effective treatment options, reshaping the landscape of cancer treatment. Hepatocellular carcinoma (HCC) is a malignant tumor originating from hepatocytes, and its pathological progression typically follows a three-step process: chronic hepatitis, cirrhosis, and cancer. Due to the insidious nature of early symptoms (such as abdominal distension and indigestion), approximately 70% of patients are diagnosed at an advanced stage, missing the opportunity for radical surgery, resulting in extremely poor overall prognosis; the 5-year survival rate for Chinese patients is only 12.1%. In 2022, there were 865,000 new cases of liver cancer globally, with 758,000 deaths, ranking third among malignant tumors in terms of mortality. In China, there were 368,000 new cases and 317,000 deaths, ranking second in mortality, with a particularly high incidence in the southeastern coastal areas. The metabolic characteristics of liver cancer are closely related to viral hepatitis (hepatitis B / C account for over 80% of cases in China), alcoholic liver disease, fatty liver, and aflatoxin exposure. These factors promote carcinogenesis by inducing chronic hepatocyte damage, DNA mutations, and metabolic reprogramming (such as enhanced glycolysis and abnormal lipid metabolism). Notably, liver cancer patients in China are more often associated with hepatitis virus infection, while those in Europe and the United States are more often associated with metabolic-related fatty liver disease. Current research progress in liver cancer drugs focuses on immunotherapy and targeted therapy. For example, atezolizumab combined with bevacizumab (the "T+A" regimen) as first-line treatment for advanced HCC can significantly improve the objective response rate (ORR) and prolong survival. In 2024, 42.54% of the innovative anti-tumor drugs approved in China were for liver cancer treatment, including PD-1 / PD-L1 inhibitors, antibody-drug conjugates (ADCs), and bispecific antibodies. However, liver cancer drug development still faces significant challenges. Most advanced-stage patients easily develop resistance to existing therapies, and tumor heterogeneity leads to low response rates to targeted therapy. Although immunotherapy has revolutionized liver cancer treatment, only a portion of patients benefit, and reliable biomarkers are lacking to guide medication. Therefore, developing highly effective and low-toxicity novel drugs (such as personalized therapies based on spatial immune characteristics) remains a top priority. Globally, the demand for liver cancer treatment continues to grow, necessitating breakthroughs in current bottlenecks through multidisciplinary collaboration and technological innovation.

[0005] Gastric cancer is a malignant tumor originating from the gastric mucosal epithelium. Its pathological progression typically involves precancerous lesions such as chronic inflammation, atrophic gastritis, intestinal metaplasia, and dysplasia, eventually developing into invasive carcinoma. Early-stage gastric cancer is confined to the mucosa or submucosa, with an annual survival rate exceeding 90%. However, the early diagnosis rate in China is less than 20%, and most patients are diagnosed at an advanced stage (invading the muscularis propria or deeper), resulting in an overall 5-year survival rate of less than 50%. In 2022, there were approximately 1.2 million new cases of gastric cancer globally, with China accounting for 40% (approximately 480,000 cases). China ranks fifth in incidence and third in mortality among malignant tumors globally, with particularly high incidence rates in Northwest and coastal areas of China. The metabolic characteristics of gastric cancer are closely related to Helicobacter pylori infection (accounting for over 80% of cases in China), high-salt diets, nitrite exposure, and metabolic reprogramming, manifesting as enhanced glycolysis, abnormal lipid metabolism, and immune microenvironment dysregulation, promoting tumor proliferation and metastasis. In recent years, significant progress has been made in gastric cancer drug research: In immunotherapy, the PD-L1 monoclonal antibody sugemalimab combined with chemotherapy as first-line treatment for advanced gastric cancer with PD-L1 CPS ≥ 5 has extended median survival to 15.6 months, becoming the first PD-L1 inhibitor approved for gastric cancer indication; in targeted therapy, antibody-drug conjugates (ADCs) and Claudin18.2-targeted therapies have shown potential, while neoadjuvant chemotherapy can shrink tumor volume and reduce the risk of postoperative metastasis. However, effective anti-gastric cancer drugs remain highly scarce, especially for advanced patients. Existing therapies are prone to failure due to tumor heterogeneity and drug resistance, and there is a lack of precise biomarkers to guide medication. Globally, although China is a leader in the field of immunotherapy combination therapy, breakthroughs are still needed in innovative therapies targeting specific molecular subtypes (such as HER2-negative or diffuse HER2), highlighting the urgent need for the development of new drugs.

[0006] Melanoma is a highly aggressive malignant tumor originating from melanocytes, commonly found in the skin, but also occurring in mucous membranes or the choroid of the eye. Its pathological progression exhibits significant heterogeneity, typically progressing from in situ lesions to aggressive metastases. Different subtypes (such as superficial spreading, nodular, and malignant lentigines) show significant differences in molecular characteristics and clinical prognosis. For example, nodular melanoma (NM) is the most aggressive, with a low 5-year survival rate, while superficial spreading melanoma (SSM) can achieve a 5-year survival rate of up to 95% when detected early. Metastatic melanoma has an extremely poor prognosis, with a 5-year mortality rate as high as 77% for patients with distant metastases. Globally, the incidence of melanoma continues to rise, with approximately 865,000 new cases and 758,000 deaths worldwide in 2022, making it the leading cause of death among skin cancers. Although the incidence rate in China is lower than in Europe and the United States, it is showing a high trend in the southeastern coastal areas due to factors such as ultraviolet radiation exposure. In terms of metabolic characteristics, melanoma often exhibits enhanced glycolysis, abnormal lipid metabolism, and immune microenvironment dysregulation. BRAF V600E mutations (approximately 50%) drive tumor progression by activating the RAS / RAF / MEK / ERK pathway. In recent years, targeted therapy and immunotherapy have significantly improved patient survival. BRAF inhibitors (such as dabrafenib) combined with MEK inhibitors (trametinib) can prolong the survival of patients with BRAF mutations, while PD-1 / CTLA-4 inhibitors (such as pembrolizumab) enhance anti-tumor effects by relieving immunosuppression. Furthermore, novel therapies such as antibody-drug conjugates (ADCs) and personalized tumor vaccines have shown potential in clinical trials. However, effective drugs remain highly scarce: only about 50% of patients carry BRAF mutations, and targeted therapy generally faces drug resistance issues; immunotherapy has an efficacy rate of only about 30%, and some subtypes (such as desmoplastic type) lack specific therapies. Drug development targeting molecular subtypes such as NRAS or NF1 mutations is still in its early stages, highlighting the urgent need for precision treatment strategies.

[0007] Therefore, developing novel broad-spectrum antitumor compounds is of great significance for researching new drugs or drug compositions for treating various cancers such as hepatocellular carcinoma. Summary of the Invention

[0008] Objective of the Invention: The objective of this invention is to provide a compound with the structure diphenylethyl 3,3'-(4,5-dihydroxy-1,2-phenylene)(2E,2'E)-diacrylate (hereinafter referred to as WX006), and to provide a class of compounds based on modified WX006, including the bulk of this compound and its pharmaceutically acceptable salts and stereoisomers.

[0009]

[0010] In some preferred embodiments, the pharmaceutically acceptable salt includes acid addition salts formed by the compound with the following acids: hydrochloric acid, hydrobromic acid, sulfuric acid, phosphoric acid, methanesulfonic acid, benzenesulfonic acid, p-toluenesulfonic acid, naphthalenesulfonic acid, citric acid, tartaric acid, lactic acid, pyruvic acid, acetic acid, maleic acid or succinic acid, fumaric acid, salicylic acid, phenylacetic acid, and mandelic acid; and also includes acid salts formed by the compound with inorganic bases.

[0011] In some preferred embodiments, the pharmaceutically acceptable salt includes basic metal cation salts, alkaline earth metal cation salts, and ammonium cation salts.

[0012] The compounds involved in this invention can also exist in the form of their salts, which are converted in vivo into the compounds mentioned in this patent. For example, within the scope of this invention, the compounds of this invention are converted into pharmaceutically acceptable salt forms according to processes known in the art, and used in salt form.

[0013] All tautomer forms of the compounds of this invention are included within the scope of this invention. The compounds of this invention may exist in specific geometric or stereoisomer forms. Additional asymmetric carbon atoms may be present in alkyl or other substituents; all such isomers and mixtures thereof are included within the scope of this invention.

[0014] WX006 and its derivatives of the present invention can be prepared by the methods described above or similar to those described above, with the appropriate starting materials selected according to the different substituents. Those skilled in the art should recognize that the above-described route helps in understanding the present invention, but does not limit the scope of the invention; unless otherwise specified, variables are defined as mentioned in general formula I.

[0015] Another object of the present invention is to provide a pharmaceutical composition comprising the compound of the present invention or a pharmaceutically acceptable salt, stereoisomer thereof, and a pharmaceutically acceptable carrier or excipient.

[0016] The pharmaceutical compositions of the present invention can be administered in various known ways, such as orally, parenterally, by inhalation spray, or via an implanted reservoir. The pharmaceutical compositions of the present invention can be administered alone or in combination with other antitumor drugs. Oral compositions can be any orally acceptable dosage form, including, but not limited to, tablets, capsules, emulsions, suspensions, dispersions, and solutions. Commonly used pharmaceutically acceptable carriers or excipients include stabilizers, diluents, surfactants, lubricants, antioxidants, binders, colorants, fillers, emulsifiers, etc.

[0017] Sterile injectable compositions may be formulated using suitable dispersants or wetting agents and suspending agents in accordance with techniques known in the art. Pharmaceutically acceptable carriers and solvents that may be used include water, mannitol, sodium chloride solution, etc.

[0018] The actual dosage level of the active ingredient in the pharmaceutical composition of the present invention can be varied to obtain an amount of active ingredient that is effective in achieving the desired therapeutic response for a particular patient, composition, and route of administration, and is non-toxic to the patient. The selected dosage level depends on a variety of factors, including the activity of the specific compound of the present invention or its salt used, the route of administration, the time of administration, the excretion rate of the specific composition used, the duration of treatment, other drugs, compounds, and / or materials used in combination with the specific composition used, the age, sex, weight, general health condition, and medical history of the patient being treated, and similar factors known in the medical field.

[0019] Another object of the present invention is to provide the use of the compounds of the present invention or pharmaceutically acceptable salts or stereoisomers thereof in the preparation of medicaments for the prevention and / or treatment of tumors.

[0020] The tumors include hepatocellular carcinoma, lung cancer, melanoma, colon cancer, skin cancer, stomach cancer, glioma, and diffuse large B-cell lymphoma.

[0021] Beneficial effects:

[0022] This invention synthesizes a class of compounds based on the modified WX006. Pharmacological experiments have demonstrated that WX006 and its derivatives possess good antitumor activity, effectively inhibiting the proliferation of various tumor cells, and show great promise in the development of antitumor drugs. Attached Figure Description

[0023] Figure 1 The hydrogen NMR spectrum of WX006;

[0024] Figure 2 The carbon NMR spectrum of WX006;

[0025] Figure 3 The IC50 value of WX006 in the experiment inhibiting Hep3B cells is... 50 ;

[0026] Figure 4 The IC50 of WX006 in the experiment inhibiting Huh7 cells is... 50 ;

[0027] Figure 5 This refers to the drug inhibition rate of WX006 on primary liver cells;

[0028] Figure 6 This is an experiment on the clonogenic effect of WX006 on Huh7 and Hep3B liver cancer cells;

[0029] Figure 7 This is a scratch assay of Hep3B and Huh7 cells treated with WX006;

[0030] Figure 8 This is the result of cell arrest caused by WX006 treatment of Hep3B and Huh7 cells;

[0031] Figure 9 It is the spleen index of the mice in Example 31;

[0032] Figure 10 It is the kidney index of the mice in Example 31;

[0033] Figure 11 It is the liver index of the mice in Example 31;

[0034] Figure 12 This refers to the tumor volume of the mouse in Example 31;

[0035] Figure 13 This refers to the body weight of the mice in Example 31;

[0036] Figure 14 These are stained sections of mouse organs from Example 31;

[0037] Figure 15 This is a quantitative fluorescence image of the tumor in mice after axillary tumor transplantation on day 0 in Example 31;

[0038] Figure 16 This refers to the body weight of the mice in Example 31 after undergoing axillary tumor implantation on day 0;

[0039] Figure 17 This refers to the quantification of tumor volume in mice after axillary tumor implantation on day 0 in Example 31;

[0040] Figure 18 This is a diagram of in situ tumor proliferation in mice from Example 31;

[0041] Figure 19 These are the tumor fluorescence quantitative images of the control group and WX006 in Example 31;

[0042] Figure 20 This is a comparison chart of the body weight of mice in the control group and WX006 in Example 31;

[0043] Figure 21 This is a comparison of tumor volume between the control group and WX006 mice in Example 31. Detailed Implementation

[0044] The preparation methods of WX006 and its derivatives of the present invention are described below with reference to specific embodiments, but these specific methods do not constitute any limitation on the present invention. The compounds of the present invention can also be conveniently prepared by optionally combining various synthetic methods described in this specification or known in the art, and such combinations can be easily performed by those skilled in the art.

[0045] The starting materials and reaction reagents used in the specific embodiments of this invention are all commercially available. This invention can be prepared into a salt form using methods commonly used in the art, such as: dissolving the compound in hydrochloric acid-ethanol at room temperature to generate hydrochloride; or adding benzenesulfonic acid to generate benzenesulfonate.

[0046] Example 1: Compound Synthesis Flow and Process

[0047] Synthesis of intermediate A:

[0048]

[0049] 4,5-Dibromo-1,2-dimethoxybenzene (30 g, 101 mmol) was dissolved in dichloromethane in a two-necked flask. Under argon protection and at -78°C, BBr3 (56 g, 222 mmol) was slowly added dropwise, and the mixture was stirred for 30 min before being brought to room temperature. After the reactants were substantially reacted as monitored by TLC, residual boron tribromide was added to quench the reaction. The mixture was extracted three times with dichloromethane, and the organic layers were washed twice with saturated sodium chloride solution. The combined organic layers were dried over anhydrous sodium sulfate and concentrated under reduced pressure to obtain a white solid, 4,5-dibromo-1,2-benzenediol, which could be directly used in the next step.

[0050] The reaction was synthesized according to the literature (HOFMANN J, FAYEZ S, SCHEINER M, et al. Sterubin: enantioresolution and configurational stability, enantiomeric purity in nature, and neuroprotective activity in vitro and in vivo[J]. Chemistry-a European Journal, 2020, 26(32): 7299-308). 4,5-Dibromo-1,2-diphenol (26.7 g, 100 mmol) was dissolved in 10 mL of acetone, and anhydrous potassium carbonate (138 g, 1 mol) was added in portions under ice bath conditions. After stirring for 1 h with a mechanical stirrer, 1-chloromethoxy-2-methoxyethane (MEMCl, 25.2 g, 202 mmol) was slowly added to the reaction system, and the reaction was carried out at room temperature for 1 h. After the reaction of the raw materials was completed by TLC monitoring, anhydrous potassium carbonate in the system was removed by vacuum filtration, acetone in the system was removed by vacuum concentration, and the product was allowed to stand for 1 hour. The product solidified from a reddish-brown oily liquid into slightly pinkish white crystals.

[0051] The above-mentioned crystals (20 g, 45 mmol), 1,3-bis(diphenylphosphine)propane (DPPP, 1.85 g, 4.5 mmol), and palladium acetate (Pd(OAc)2, 1 g, 4.5 mmol) were placed in a Schlenk tube and uniformly dispersed in 100 mL of 1,4-dioxane. Triethylamine (18 g, 180 mmol) and ethyl acrylate (45 g, 450 mmol) were added sequentially. Under argon protection, the reaction was heated at 120 °C for 48 h. After monitoring for complete reaction, the system was cooled to room temperature, and insoluble matter was removed by filtration. The filtrate was diluted with ethyl acetate. The organic phase was washed sequentially with saturated sodium chloride solution, saturated sodium bicarbonate solution, and saturated ammonium chloride solution. The resulting organic phase was dried over anhydrous sodium sulfate and concentrated under reduced pressure. The crude product was a reddish-brown oily liquid and could be directly added to the next reaction step.

[0052] The above oily liquid was dissolved in 20 ml of ethanol, and potassium hydroxide (7.7 g, 135 mmol) was added. The mixture was stirred thoroughly, and the reaction was monitored by TLC until it was complete. The reaction solution was diluted with water, and the pH was adjusted to 2 with 1 N HCl under ice bath. The mixture was extracted three times with ethyl acetate, and the organic phase was extracted four times with saturated sodium bicarbonate solution. The product was converted to carboxylate and back-extracted into the aqueous phase. The aqueous phases were combined, and the pH was adjusted to 2 under ice bath. The mixture was extracted three times with ethyl acetate again. The organic phases were combined, washed three times with saturated sodium chloride solution, dried with anhydrous sodium sulfate, and concentrated under reduced pressure to obtain a brownish-gray solid, which was compound A. The overall yield of the four-step reaction was 90%.

[0053] Intermediate (A) is a grayish-white powder (90%). 1 H NMR (300MHz, Methanol-d4) δ8.04(d,J=15.7Hz,1H),7.52(s,1H),6.39(d,J=15.7Hz,1H),5.38(s,2H),3.90(t,2H),3.62(t,2H),3.38(s,3H).

[0054] General method for synthesizing compounds (Examples 1-19):

[0055] Referring to the literature (WANG YH, LIQ-S, WANG PL, et al. Synthesis of Caffeic Acid Phenethyl Ester Analogues and Their Cytotoxicities Against Human Cancer Cells[J]. Asian Journal of Chemistry, 2014, 26(9): 2686-90), compound (A) (426 mg, 1 mmol) and substituted aromatic alcohol (2.2 mmol) were dissolved in 2 mL of dichloromethane (DCM), and condensing agent (1-ethyl-3-(3-dimethylaminopropylamine)carbodiimide) (EDCl, 1.53 g, 8 mmol) and 4-dimethylaminopyridine (DMAP, 268 mg, 2.2 mmol) were added. The mixture was stirred at room temperature for 1 h under nitrogen protection. After the reaction of the starting materials was completed as monitored by TLC, the dichloromethane was evaporated to dryness, diluted with water, and extracted three times with ethyl acetate. The organic phases were combined, washed three times with saturated sodium chloride solution, dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The crude product was subjected to silica gel column chromatography (petroleum ether: ethyl acetate = 10:1) to give a brown oily substance in yield of 61-70%.

[0056] Referring to the literature (OHYOSHI T, MITSUGI K, ICHIMRA F, et al. Total Synthesis and Structure-Activity Relationship Studies of Phelligridins C and D, and Phellifuropyranone A[J]. Bulletin of the Chemical Society of Japan, 2020, 93(12): 1540-51), the above compound (1 eq) was dissolved in 1 mL of methanol, and then 4N HCl (1 eq) was added, and the mixture was refluxed at 70 °C for 2 h. After the reaction was basically completed as monitored by TLC, the reaction solution was directly evaporated to dryness to obtain a light yellow solid. The solid was extracted with ethyl acetate, and the organic layer was washed twice with saturated sodium chloride solution. The organic layers were combined, dried with anhydrous sodium sulfate, and concentrated under reduced pressure. The crude product was subjected to silica gel column chromatography (petroleum ether: ethyl acetate = 4:1) to obtain a light yellow solid (Examples 1-19), with a yield of 62-68%.

[0057] The synthesis method of WX006:

[0058] Referring to the literature [WANG YH, LIQ-S, WANG PL, et al. Synthesis of Caffeic Acid Phenethyl Ester Analogues and Their Cytotoxicities Against Human Cancer Cells[J]. Asian Journal of Chemistry, 2014, 26(9): 2686-90.], compound (A) (426 mg, 1 mmol) and phenylethanol (2.2 mmol) were dissolved in 2 mL of dichloromethane (DCM), and condensing agents (1-ethyl-3-(3-dimethylaminopropylamine)carbodiimide) (EDCl, 1.53 g, 8 mmol) and 4-dimethylaminopyridine (DMAP, 268 mg, 2.2 mmol) were added. The mixture was stirred at room temperature for 0.5-5 h under nitrogen protection. After the reaction of the starting materials was completed as monitored by TLC, the dichloromethane was evaporated to dryness, diluted with water, and extracted three times with ethyl acetate. The organic phases were combined, washed three times with saturated sodium chloride solution, dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The crude product was subjected to silica gel column chromatography (petroleum ether: ethyl acetate = 10:1) to give a brown oily substance in 65% yield.

[0059] Referring to the literature [OHYOSHI T, MITSUGI K, ICHIMURA F, et al. Total Synthesis and Structure-Activity Relationship Studies of Phelligridins C and D, and Phellifuropyranone A[J]. Bulletin of the Chemical Society of Japan, 2020, 93(12): 1540-51.], the above crude product (1 eq) was dissolved in 1 mL of methanol, and then 4N HCl (1 eq) was added, and the mixture was refluxed at 70 °C for 2 h. After the reaction of the raw materials was basically completed as monitored by TLC, the reaction solution was directly evaporated to dryness to obtain a light yellow solid. The product was extracted with ethyl acetate, the organic layer was washed twice with saturated sodium chloride solution, the organic layers were combined and dried with anhydrous sodium sulfate, and concentrated under reduced pressure. The crude product was subjected to silica gel column chromatography (petroleum ether: ethyl acetate = 4:1) to obtain a light yellow solid (WX006), with a yield of 40%. Figure 1 , 2 These are the hydrogen and carbon NMR spectra of WX006.

[0060] (Diphenylethyl 3,3'-(4,5-dihydroxy-1,2-phenylene)(2E,2'E)-diacrylate)

[0061] Pale yellow solid (40%); melting point (mp), 139-144℃; 1 H NMR (300MHz, Methanol-d4) δ7.99(d,J=15.6Hz,1H),7.33-7.17(m,5H),7.10(s,1H),6.22(d,J=15.6Hz,1H),4.39(t,J=6.9Hz,2H),3.00(t,J=6.9Hz,2H); 13 C NMR(75MHz, Methanol-d4)δ167.11,148.07,140.69,137.98,128.65,128.17,126.56,126.19,117.44,112.88,65.03,34.79; MS(ESI)m / z,459.17[M+H] + .

[0062] Example 2: (bis(3-phenylpropyl)3,3'-(4,5-dihydroxy-1,2-phenylene)(2E,2'E)-diacrylate)

[0063] Pale yellow solid (36.5%); melting point (mp), 138-144℃;1 H NMR (300MHz, Acetone-d6) δ8.78 (s, 2H), 8.05 (d, J = 15.7Hz, 2H), 7.36-7.13 (m, 12H), 6.3 2(d,J=15.6Hz,2H),4.21(t,J=6.5Hz,4H),3.76(s,0H),2.77(t,4H),2.11-1.95(m,8H); 13 C NMR (75MHz, Acetone-d6) δ166.14,147.79,141.51,140.30,128.41,128.35,12 6.92,125.85,118.53,113.51,63.41,31.91,30.39; MS(ESI)m / z,487.21[M+H] + .

[0064] Example 3: (bis(4-phenylbutyl)3,3'-(4,5-dihydroxy-1,2-phenylene)(2E,2'E)-diacrylate)

[0065] Pale yellow solid (37.7%); melting point (mp), 140-146℃; 1 H NMR (300MHz, Acetone-d6) δ8.80 (s, 2H), 8.03 (d, J = 15.7Hz, 2H), 7.35-7.12 ( m,12H),6.30(d,J=15.7Hz,2H),4.23(t,4H),2.71(t,4H),1.89-1.55(m,8H); 13 C NMR (75MHz, Acetone-d6) δ166.18,147.79,142.24,140.24,128.36,128.25,126.8 8,125.68,118.54,113.47,63.90,35.16,28.26,27.78; MS(ESI)m / z,515.24[M+H] + .

[0066] Example 4: (bis(4-methylphenylethyl)3,3'-(4,5-dihydroxy-1,2-phenylene)(2E,2'E)-diacrylate)

[0067] Pale yellow solid (38.2%); melting point (mp), 180-183℃; 1HNMR(300MHz, Acetone-d6)δ8.85(s,1H),8.04(d,J=15.7Hz,2H),7.27(s,2H),7.26-7.12(m,8H) ,6.30(d,J=15.6Hz,2H),4.39(t,J=6.9Hz,4H),3.79(s,0H),2.99(t,J=6.9Hz,4H),2.30(s,6H). 13 C NMR (75MHz, Acetone-d6) δ166.12,147.76,140.28,135.73,135.22,129.10,12 8.92,126.90,118.44,113.41,65.01,34.55,20.23; MS(ESI)m / z,587.20[M+H] + .

[0068] Example 5: (bis(3-(p-tolyl)propyl)3,3'-(4,5-dihydroxy-1,2-phenylene)(2E,2'E)-diacrylate)

[0069] Pale yellow solid (37.6%); melting point (mp), 142-145℃; 1 H NMR (300MHz, Methanol-d4) δ8.01(d,J=15.6Hz,2H),7.12(d,J=8.6Hz,10H),6.27(d,J=15 .6Hz,2H),3.38-3.32(m,14H),2.72(t,J=7.6Hz,4H),2.29(s,7H),2.03(q,J=7.0Hz,5H); 13 C NMR (75MHz, Acetone-d6) δ166.18,147.75,140.28,138.41,129.01,128.35,1 26.93,118.54,113.47,63.47,31.51,30.49,20.18; MS(ESI)m / z,601.20[M+H] + .

[0070] Example 6: (bis(4-(p-tolyl)butyl)3,3'-(4,5-dihydroxy-1,2-phenylene)(2E,2'E)-diacrylate)

[0071] Pale yellow solid (36.8%); melting point (mp), 142-145℃; 1HNMR (300MHz, Acetone-d6) δ8.87(s,1H),8.04(d,J=15.7Hz,2H),7.27(s,2H),7.13(q,J=5.7Hz,8H),6.31( d,J=15.7Hz,2H),5.66(s,1H),4.23(t,J=5.8Hz,4H),2.67(t,J=6.4Hz,4H),2.29(s,6H),1.79-1.69(m,8H); 13 C NMR (75MHz, Acetone-d6) δ166.22,140.23,139.13,134.89,128.91,128.31,1 26.90,118.55,113.43,63.96,34.76,27.90,20.17; MS(ESI)m / z,615.20[M+H] + .

[0072] Example 7: (bis(2-chlorophenylethyl)3,3'-(4,5-dihydroxy-1,2-phenylene)(2E,2'E)-diacrylate)

[0073] Pale yellow solid (39.6%); melting point (mp), 152-160℃; 1 HNMR (300MHz, Acetone-d6) δ9.00 (s, 1H), 8.04 (d, J = 15.7Hz, 2H), 7.70-7.10 ( m,10H),6.29(d,J=15.6Hz,2H),4.46(t,J=6.9Hz,4H),3.22(d,J=7.1Hz,4H); 13 C NMR (75MHz, Acetone-d6) δ166.09,147.90,140.42,135.85,133.80,131.63,129.46 ,128.46,127.30,126.82,118.21,113.33,63.13,32.62; MS(ESI)m / z,527.10[M+H] + .

[0074] Example 8: (bis(3-chlorophenylethyl)3,3'-(4,5-dihydroxy-1,2-phenylene)(2E,2'E)-diacrylate)

[0075] Pale yellow solid (44.1%); melting point (mp), 142-146℃; 1HNMR (300MHz, Acetone-d6) δ8.98 (s, 1H), 8.03 (d, J = 15.7Hz, 2H), 7.44-7.23 ( m,10H),6.30(d,J=15.6Hz,2H),4.44(t,J=6.8Hz,4H),3.06(t,J=6.7Hz,4H); 13 C NMR (75MHz, Acetone-d6) δ166.12,147.90,140.99,140.45,133.70,130.13,128.98 ,127.72,126.80,126.55,118.17,113.35,64.45,34.51; MS(ESI)m / z,527.10[M+H] + .

[0076] Example 9: (bis(4-chlorophenylethyl)3,3'-(4,5-dihydroxy-1,2-phenylene)(2E,2'E)-diacrylate)

[0077] Pale yellow solid (41.5%); melting point (mp), 190-194℃; 1 H NMR (300MHz, Acetone-d6) δ8.91 (s, -1H), 8.02 (d, J = 15.5Hz, 2H), 7.41-7.33 (m, 8H) ,7.26(s,2H),6.29(d,J=15.6Hz,2H),4.42(t,J=6.6Hz,4H),3.05(t,J=6.5Hz,4H); 13 CNMR(75MHz,Acetone-d6)δ166.12,147.89,140.37,137.47,131.79,130.80,128.46,126.80,118.21,113.35,64.58,34.22; MS(ESI)m / z,527.10[M+H] +

[0078] Example 10: (bis(3-(4-chlorophenyl)propyl)3,3'-(4,5-dihydroxy-1,2-phenylene)(2E,2'E)-diacrylate)

[0079] Pale yellow solid (39.3%); melting point (mp), 150-153℃; 1H NMR (300MHz, Acetone-d6) δ8.06(d,J=15.8Hz,2H),6.33(d,J=15.9Hz,2H),4.22(t,4H),2.77(t,J=6.9Hz,4H),2.07-1.91(m,4H); 13 C NMR (75MHz, Acetone-d6) δ166.21,147.74,140.50,140.37,131.19,130.22,12 8.39,126.94,118.45,113.47,63.33,31.27,30.27; MS(ESI)m / z,555.15[M+H] + .

[0080] Example 11: (bis[3-(4-chlorophenyl)propyl]3,3'-(4,5-dihydroxy-1,2-phenylene)(2E,2'E)-diacrylate)

[0081] Pale yellow solid (38.2%); melting point (mp), 160-165℃; 1 H NMR (300MHz, Acetone-d6) δ8.04(d,J=15.5Hz,2H),7.31(q,J=8.7Hz,10H),6.32(d, J=15.6Hz,2H),4.24(d,J=5.7Hz,4H),2.71(t,J=6.9Hz,4H),1.76(p,J=3.3Hz,8H); 13 C NMR (75MHz, Acetone-d6) δ166.24,147.91,141.25,140.27,131.33,130.18,128.2 9,126.83,118.44,113.42,63.86,34.45,28.20,27.71; MS(ESI)m / z,583.14[M+H] + .

[0082] Example 12: (bis[4-(trifluoromethyl)phenethyl]3,3'-(4,5-dihydroxy-1,2-phenylene)(2E,2'E)-diacrylate)

[0083] Pale yellow solid (42.1%); melting point (mp), 179-183℃; 1H NMR(300MHz, Methanol-d4)δ8.02(d,J=15.7Hz,2H),7.79-7.34(m,8H),7.12(s,2H),6.27( d,J=15.6Hz,2H),4.22(t,J=6.4Hz,4H),2.84(t,J=6.4Hz,4H),2.22-1.90(q,J=6.4Hz,3H); 13 C NMR (75MHz, Acetone-d6) δ166.12,147.91,143.43,140.43,129.81,126.79,125. 31(q,J=5.6Hz),125.27,118.14,113.33,64.30,34.68; MS(ESI)m / z,594.15[M+H] +

[0084] Example 13: (bis[3-(4-trifluoromethylphenyl)propyl]3,3'-(4,5-dihydroxy-1,2-phenylene)(2E,2'E)-diacrylate)

[0085] Pale yellow solid (40.6%); melting point (mp), 136-144℃; 1 H NMR (300MHz, Acetone-d6) δ8.06(d,J=15.8Hz,2H),6.33(d,J=15.9Hz,2H),4.22(t,4H),2.77(t,J=6.9Hz,4H),2.07-1.91(m,4H); 13 C NMR (75MHz, Acetone-d6) δ166.21,147.74,140.50,140.37,131.19,130.22,128.39 ,126.94(q,J=5.6Hz),118.45,113.47,63.33,31.27,30.27; MS(ESI)m / z,608[M+H] + .

[0086] Example 14: (bis(2,3-dichlorophenylethyl)3,3'-(4,5-dihydroxy-1,2-phenylene)(2E,2'E)-diacrylate)

[0087] Pale yellow solid, 40.0%; melting point (mp), 192-198℃; 1H NMR (300MHz, Acetone-d6) δ8.01(d,J=15.7Hz,2H),7.56-7.30(m,6H),7.24(s,2H),6.28(d,J=15.7Hz,2H),4.49(t,J=6.6Hz,4H),3.27(t,J=6.6Hz,4H); 13 C NMR (75MHz, Acetone-d6) δ166.02,147.89,140.49,138.63,132.58,131.89,130.06 ,129.05,127.95,126.80,118.13,113.34,62.74,33.53; MS(ESI)m / z,345.09[M+H] + .

[0088] Example 15: (bis(2,4-dichlorophenylethyl)3,3'-(4,5-dihydroxy-1,2-phenylene)(2E,2'E)-diacrylate)

[0089] Pale yellow solid (41.7%); melting point (mp), 147-150℃; 1 H NMR (300MHz, Acetone-d6) δ8.94 (s, 1H), 8.02 (d, J = 15.6Hz, 2H), 7.45 (d, J = 34.3 Hz,5H),7.25(s,2H),6.27(d,J=15.2Hz,2H),4.46(s,4H),3.20(d,J=6.8Hz,3H); 13 C NMR (75MHz, Acetone-d6) δ166.06,147.91,140.48,135.06,134.69,132.72,12 8.96,127.43,126.82,118.12,113.35,62.90,32.06; MS(ESI)m / z,596.01[M+H] + .

[0090] Example 16: (bis(2,5-dichlorophenylethyl)3,3'-(4,5-dihydroxy-1,2-phenylene)(2E,2'E)-diacrylate)

[0091] Pale yellow solid (43.2%); melting point (mp), 164-170℃; 1H NMR(300MHz, Acetone-d6)δ7.97(d,J=15.7Hz,2H),7.51-7.38(m,4H),7.28(dd,J=8.6,2.5 Hz,2H),7.20(s,2H),6.23(d,J=15.7Hz,2H),4.43(t,J=6.6Hz,4H),3.16(t,J=6.6Hz,4H); 13 C NMR (75MHz, Acetone-d6) δ166.02,147.91,140.62,138.13,132.51,132.36,131.14 ,130.88,128.33,126.84,118.14,113.40,62.84,32.48; MS(ESI)m / z,596.01[M+H] + .

[0092] Example 17: (bis(2,6-dichlorophenylethyl)3,3'-(4,5-dihydroxy-1,2-phenylene)(2E,2'E)-diacrylate)

[0093] Pale yellow solid (39.6%); melting point (mp), 132-136℃; 1 H NMR (300MHz, Acetone-d6) δ8.96 (s, 1H), 8.04 (d, J = 15.7Hz, 2H), 7.46 (d, J = 8.1Hz, 4H), 7.38-7 .23(m,4H),6.28(d,J=15.7Hz,2H),5.65(s,1H),4.48(t,J=6.7Hz,4H),3.41(t,J=6.5Hz,5H); 13 C NMR(75MHz, Acetone-d6)δ166.02,147.89,140.54,135.69,133.95,129.14,128.58,126.92,118.27,113.38,61.80,30.47; MS(ESI)m / z,596.01[M+H] + .

[0094] Example 18: (bis(3,4-dichlorophenylethyl)3,3'-(4,5-dihydroxy-1,2-phenylene)(2E,2'E)-diacrylate)

[0095] Pale yellow solid (37.2%); melting point (mp), 192-198℃; 1H NMR(300MHz, Acetone-d6)δ8.90(s,2H),8.03(dd,J=15.7,2.7Hz,2H),7.62-7.51(m,4H),7.37(dd,J=8.3 ,2.1Hz,2H),7.26(d,J=2.6Hz,2H),6.30(d,J=15.6Hz,2H),4.45(t,J=6.6Hz,4H),3.08(t,J=6.5Hz,8H); 13 C NMR (75MHz, Acetone-d6) δ166.04,147.90,140.52,139.73,131.64,131.08,130.51 ,129.82,129.34,126.84,118.17,113.46,64.19,33.96; MS(ESI)m / z,596.01[M+H] +

[0096] Example 19: (bis(3,5-dichlorophenylethyl)3,3'-(4,5-dihydroxy-1,2-phenylene)(2E,2'E)-diacrylate)

[0097] Pale yellow solid (36.5%); melting point (mp), 202-206℃; 1 H NMR(300MHz, Acetone-d6)δ8.03(d,J=15.6Hz,2H),7.39(dd,J=8.2,2.0Hz,6H),7 .26(s,2H),6.30(d,J=15.7Hz,2H),4.46(t,J=6.6Hz,4H),3.09(t,J=6.6Hz,4H); 13 C NMR(75MHz, Acetone-d6)δ165.99,147.88,142.82,140.62,134.51,127.85,126.87,126.36,118.17,113.52,63.98,34.26; MS(ESI)m / z,596.01[M+H] + .

[0098] Example 20: General method for synthesizing compounds (Examples 20-28):

[0099] Substituted 3-phenyl-2-propanols (2.2 mmol) and triphenylphosphine (557 mg, 2.2 mmol) of different chiralities were dissolved in 6 mL of tetrahydrofuran (THF). Diisopropyl azodicarbonate (DIAD, 444 mg, 2.2 mmol) was added under ice bath conditions, and the mixture was stirred for 15 min. Then, intermediate A (426 mg, 1 mmol) was dissolved in 2 mL of tetrahydrofuran (THF), and the resulting solution was added dropwise to the aforementioned reaction solution. After the starting material was completely reacted as monitored by TLC, the reaction was quenched with water, and the mixture was extracted three times with ethyl acetate. The organic phases were combined, washed with saturated sodium chloride solution, dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The crude product was subjected to silica gel column chromatography (petroleum ether: ethyl acetate = 6:1) to give a white solid in yield of 72-78%.

[0100] Referring to the literature, the above compound (1 eq) was dissolved in 1 mL of methanol, and then 4N HCl (4 eq) was added. The mixture was refluxed at 70 °C for 2 h. After the reactants were basically reacted as monitored by TLC, the reaction solution was directly evaporated to dryness to obtain a pale yellow solid. The solid was extracted with ethyl acetate, and the organic layer was washed twice with saturated sodium chloride solution. The combined organic layers were dried over anhydrous sodium sulfate and concentrated under reduced pressure. The crude product was subjected to silica gel column chromatography (petroleum ether: ethyl acetate = 4:1) to obtain a pale yellow solid (Examples 20-28), with a yield of 61-68%.

[0101] (bis[(R)-1-phenylpropyl-2-yl]3,3'-(4,5-dihydroxy-1,2-phenylene)(2E,2'E)-diacrylate)

[0102] Pale yellow solid (45.2%); melting point (mp), 120-124℃; ¹H NMR (300MHz, Acetone-d6) δ 8.76 (s, 1H), 7.99 (d, J = 15.7Hz, 1H), 7.35-7.24 (m, 4H), 7.22 (s, 1H), 6.24 (d, J = 15.7Hz, 1H), 5.29-5.12 (m, 1H), 3.04-2.84 (m, 2H), 1.28 (d, J = 6.2Hz, 3H); ¹³C NMR (75MHz, Acetone-d6) δ166.50,148.66,140.91,138.90,130.34,129.14,127 .73,127.21,119.67,114.26,72.34,42.78,19.90; MS(ESI)m / z,487.20[M+H]+.

[0103] Example 21: (bis[(S)-1-phenylprop-2-yl]3,3'-(4,5-dihydroxy-1,2-phenylene)(2E,2'E)-diacrylate)

[0104] Pale yellow solid (42.8%); melting point (mp), 120-124℃; 1 HNMR (300MHz, Acetone-d6) δ8.77 (s, 1H), 8.01 (d, J = 15.7Hz, 2H), 7.37-7.26 (m, 10H), 7.24(s,2H),6.26(d,J=15.7Hz,2H),5.22(h,2H),2.95(h,4H),1.30(d,J=6.2Hz,6H); 13 C NMR (75MHz, Acetone-d6) δ165.63,147.80,140.04,138.04,129.47,128.28,12 6.87,126.35,118.81,113.40,71.47,41.92,19.04; MS(ESI)m / z,487.20[M+H] + .

[0105] Example 22: (bis(1-phenylprop-2-yl)3,3'-(4,5-dihydroxy-1,2-phenylene)(2E,2'E)-diacrylate)

[0106] Pale yellow, waxy substance (48.3%); 1 H NMR (300MHz, Acetone-d6) δ8.76 (s, 2H), 8.00 (d, J = 15.7Hz, 2H), 7.36-7.25 (m, 8H), 7.23 (s ,2H),6.25(d,J=15.7Hz,2H),5.29-5.13(m,2H),3.04-2.85(m,4H),1.29(d,J=6.2Hz,6H); 13 C NMR (75MHz, Acetone-d6) δ166.52,148.68,140.93,138.93,130.36,129.17,127 .75,127.23,119.70,114.29,72.36,42.81,19.93; MS(ESI)m / z,487.20[M+H]+.

[0107] Example 23: (bis[(R)-1-(p-tolyl)propyl-2-yl]3,3'-(4,5-dihydroxy-1,2-phenylene)(2E,2'E)-diacrylate)

[0108] Pale yellow solid (43.9%); melting point (mp), 143-148℃; 1HNMR(300MHz,Methanol-d4)δ7.97(d,J=15.7Hz,2H),7.49-6.91(m,10H),6.19(d,J=15.6 Hz,2H),5.17(h,J=5.7,5.1Hz,2H),2.79-2.99(m,4H),2.27(s,6H),1.27(d,J=5.7Hz,6H); 13 C NMR (75MHz, Methanol-d4) δ166.69,148.07,140.47,135.71,134.55,129.03,128. 63,126.54,117.92,112.82,72.00,41.41,19.71,18.49; MS(ESI)m / z,524.24[M+H] + .

[0109] Example 24: (bis[(S)-1-(p-Tolyl)propyl-2-yl]3,3'-(4,5-dihydroxy-1,2-phenylene)(2E,2'E)-diacrylate)

[0110] Pale yellow solid (41.1%); melting point (mp), 173-148℃; 1 HNMR(300MHz,Methanol-d4)δ7.98(d,J=15.7,2H),7.12(m,10H),6.20(d,J=15.6, 2H),5.18(h,J=5.7,5.1,2H),2.79-2.99(m,2H),2.29(s,6H)1.28(d,J=5.7Hz,6H); 13 C NMR (75MHz, Methanol-d4) δ166.69,148.06,140.47,135.70,134.54,129.03,128. 63,126.54,117.92,112.83,72.00,41.41,19.73,18.49; MS(ESI)m / z,524.24[M+H] + .

[0111] Example 25: (bis[1-(p-tolyl)propyl-2-yl]3,3'-(4,5-dihydroxy-1,2-phenylene)(2E,2'E)-diacrylate)

[0112] Pale yellow, waxy substance (48.6%); 1H NMR (300MHz, Acetone-d6) δ8.77 (s, 1H), 8.01 (d, J = 15.7Hz, 2H), 7.37-7.26 (m, 10H), 7.24(s,2H),6.26(d,J=15.7Hz,2H),5.22(h,2H),2.95(h,4H),1.30(d,J=6.2Hz,6H); 13 C NMR (75MHz, Acetone-d6) δ165.63,147.80,140.04,138.04,129.47,128.28,126.8 7,126.35,118.81,113.40,71.47,41.92,20.01,19.04; MS(ESI)m / z,524.24[M+H] + .

[0113] Example 26: (bis[(R)-1-(4-chlorophenyl)propyl-2-yl]3,3'-(4,5-dihydroxy-1,2-phenylene)(2E,2'E)-diacrylate)

[0114] Pale yellow solid (44.1%); melting point (mp), 173-176℃; 1 H NMR (300MHz, Methanol-d4) δ7.99 (d, J=15.6Hz, 2H), 7.32-7.24 (m, 8H), 7.11 (s ,2H),6.22(d,J=15.6Hz,2H),5.23(m,2H),2.94(m,4H),1.31(d,J=6.3Hz,6H); 13 C NMR (75MHz, Acetone-d6) δ165.65,147.81,140.15,137.01,131.74,131.25,12 8.29,126.76,118.59,113.30,71.19,41.07,19.02; MS(ESI)m / z,556.13[M+2H] + .

[0115] Example 27: (bis[(S)-1-(4-chlorophenyl)propyl-2-yl]3,3'-(4,5-dihydroxy-1,2-phenylene)(2E,2'E)-diacrylate)

[0116] Pale yellow solid (44.6%); melting point (mp), 173-176℃; 1H NMR (300MHz, Methanol-d4) δ7.99 (d, J=15.6Hz, 2H), 7.32-7.24 (m, 8H), 7.11 (s ,2H),6.22(d,J=15.6Hz,2H),5.23(m,2H),2.94(m,4H),1.31(d,J=6.3Hz,6H); 13 C NMR(75MHz,Chloroform-d)δ166.64,148.12,140.66,136.54,132.05,130.81,1 28.11,126.58,117.85,112.94,71.59,41.06,18.56; MS(ESI)m / z,556.13[M+2H] + .

[0117] Example 28: (bis[1-(4-chlorophenyl)prop-2-yl]3,3'-(4,5-dihydroxy-1,2-phenylene)(2E,2'E)-diacrylate)

[0118] Pale yellow, waxy substance (40.3%); 1H NMR (300MHz, Acetone-d6) δ 8.77 (s, 1H), 8.01 (d, J = 15.7Hz, 2H), 7.37–7.26 (m, 10H), 7.24 (s, 2H), 6.26 (d, J = 15.7Hz, 2H), 5.22 (h, 2H), 2.95 (h, 4H), 1.30 (d, J = 6.2Hz, 6H); 13C NMR (75MHz, Acetone-d6) δ165.63,147.80,140.04,138.04,129.47,128.28,126 .87,126.35,118.81,113.40,71.47,41.92,19.04; MS(ESI)m / z,556.13[M+2H]+.

[0119] Example 29: Assay for biological activity

[0120] Experimental method: 10 mg of the different compounds synthesized in the above examples were accurately weighed, dissolved in anhydrous DMSO, and prepared into 100 mM stock solutions.

[0121] For adherent cells Hep3B, Huh7, HepG2, B16-F10, H1299, H1975, HCT-116, U87, U251, SHSY5Y, AGS, MKN45, A431, SCC13, Farage, and MC116, cells were spotted into 96-well plates at a density of 3000 cells / well and 100 μL per well. The corresponding small molecule stock solutions were prepared as 200 μM serially diluted 3-fold gradient drug-containing media. After cell adhesion, the original medium was discarded. (For suspension cells Farage and MC116, a half-volume media replacement method was used, increasing the initial cell count to 6000 cells / well). 100 μL of the serially diluted working solution was added to each well of the culture plate containing the seeded cells. After 48 hours of culture, absorbance was measured using the CCK8 assay, and IC50 was calculated using Graphpad Prism. 50 value.

[0122] The experimental results are shown in Table 1.

[0123] Table 1. IC50 of the antiproliferative activity against 7 cancer cell lines in the examples. 50 Value (μM)

[0124]

[0125]

[0126]

[0127] Brief introduction of cell lines used to detect compound activity

[0128] Hep3B cells are a liver cancer cell line that is widely used in liver cancer research.

[0129] Huh7 originated from a male with well-differentiated hepatocellular carcinoma and is the only cell line capable of effectively replicating the hepatitis C virus.

[0130] HepG2 cells were derived from liver cancer tissue of a 15-year-old girl and are suitable for research on liver cell metabolism.

[0131] B16-F10 cells are derived from melanoma tissue of C57BL / 6J mouse skin and are a subline of the B16 tumor cell line. They are widely used to study the formation, invasion and metastasis of tumor cells.

[0132] H1299 cells were derived from lymph node metastases in a 43-year-old male patient and are widely used in lung cancer research; H1975 cells were isolated from non-small cell lung adenocarcinoma tissue in a non-smoking woman and are mainly used in lung cancer research and immuno-oncology research.

[0133] HCT-116 was isolated from the lymph nodes of a 51-year-old male patient with colon cancer and has been widely used in colorectal cancer and toxicology studies.

[0134] U87-MG is a widely used human glioblastoma (GBM) cell line, originally established in 1966 by Uppsala University in Sweden from glioblastoma tissue of a 50-year-old female patient. This cell line was later indexed by the American Type Culture Collection (ATCC) (HTB-14) and is primarily used to test the in vitro efficacy of chemotherapeutic drugs (such as temozolomide), targeted therapies (such as EGFR inhibitors), and immunotherapies (such as PD-1 antibodies).

[0135] U251 cells (also known as U-251MG) are a human glioblastoma (GBM) cell line, initially established in 1966 from glioblastoma tissue of a 75-year-old male patient. This cell line is widely used in neuro-oncology research.

[0136] SH-SY5Y is a human neuroblastoma cell line, established in 1970 from bone marrow metastases in a 4-year-old girl. This cell line is widely used in neurobiology, oncology, and research on neurodegenerative diseases.

[0137] AGS cells are a human gastric adenocarcinoma cell line, initially established in 1983 by Barranco et al. from untreated gastric adenocarcinoma resection tissue of a 54-year-old female patient. This cell line is indexed by the American Type Culture Collection (ATCC) (CRL-1739) and is widely used in research on the pathogenesis, drug screening, and molecular biology of gastric cancer.

[0138] MKN45 cells are a human gastric adenocarcinoma cell line, originally isolated and established by S. Akiyama or Hojo H (different sources in different literature) from a poorly differentiated gastric adenocarcinoma liver metastasis in a 62-year-old female patient. They are mainly used to explore the mechanisms of tumor proliferation, invasion, metastasis and drug resistance.

[0139] A431 cells are a human epidermal carcinoma cell line that was isolated and established in 1973 by Giard et al. from epidermal carcinoma tissue of an 85-year-old female patient. They are widely used in cancer research, drug development and signal transduction research.

[0140] SCC13 cells are a human cutaneous squamous cell carcinoma (cSCC) cell line, initially isolated from tumor tissue of a patient with cutaneous squamous cell carcinoma. This cell line is widely used in research on the pathogenesis of skin cancer, drug screening, and the tumor microenvironment.

[0141] SCC13 cells retain the typical characteristics of squamous cell carcinoma, including abnormal EGFR signaling pathway and invasive growth characteristics, making them an important in vitro model for studying skin squamous cell carcinoma.

[0142] Farage cells are a human diffuse large B-cell lymphoma (DLBCL) cell line established in 1990 from a lymph node biopsy of an adult female patient by the H. Ben-Bassat laboratory. This cell line is indexed by the American Type Culture Collection (ATCC) under accession number CRL-2630 and is primarily used to study the proliferation, drug resistance, and signaling pathway mechanisms of DLBCL.

[0143] MC116 cells are a human undifferentiated lymphoma cell line established by M. Romsdahl's laboratory and isolated from Dukes' C-type colorectal adenocarcinoma (grade IV) tissue from a male patient. This cell line is indexed by the American Type Culture Collection (ATCC) under the number CRL-1649 and is widely used in lymphoma biology, immunotherapy, and drug screening research.

[0144] Table 1 shows that WX006 and its derivatives can effectively inhibit the in vitro growth of hepatocellular carcinoma, lung cancer, colon cancer, melanoma, skin cancer, gastric cancer, glioma, and lymphoma cell lines.

[0145] The above experimental results show that the compound provided by the present invention can effectively inhibit the in vitro growth of hepatocellular carcinoma, lung cancer, colon cancer, melanoma, skin cancer, gastric cancer, glioma, and lymphoma cell lines.

[0146] The compounds of the present invention and their medically acceptable salts can effectively inhibit the in vitro growth of hepatocellular carcinoma, lung cancer, colon cancer, melanoma, skin cancer, gastric cancer, glioma, and lymphoma cell lines, and can be used as active ingredients in pharmaceuticals. Therefore, drugs containing the above compounds as active ingredients can be used to prepare drugs for the prevention and / or treatment of tumors.

[0147] Example 30

[0148] The tumor-suppressive effect of WX006 in vitro was detected using CCK8 assay, colony assay, scratch assay, and Western blotting.

[0149] The specific steps are as follows:

[0150] CCK8 assay for cell viability

[0151] The CCK-8 (Cell Counting Kit-8) reagent contains WST-8 (water-soluble tetrazolium salt). This reagent can be reduced by dehydrogenases in cells to highly water-soluble orange formazan under the action of electron carrier 1-Methoxy PMS. The amount of formazan generated is directly proportional to the number of live cells. Therefore, the absorbance value measured by an ELISA reader at a wavelength of 450 nm can indirectly reflect the number of live cells. This patent study uses this method to detect cell proliferation and cytotoxicity.

[0152] Stable tumor cells with a confluence of 70% were seeded at a concentration of 3000 cells / 100 μL into 96-well flat-bottom cell culture plates and incubated at 37°C in a 5% CO2 incubator for 24 h. After cell attachment, the original culture medium was discarded, and 100 μL of culture medium containing different concentrations of the compound from the examples, or the target compound composition, or a blank medium containing an equal amount of DMSO was added as soon as possible. After incubation with the drug or drug composition for 24 or 48 hours, the drug treatment working solution was discarded, and 100 μL of CCK8 working solution containing 10% CCK-8 stock solution was added to each well. The cells were incubated for 3.5 h. After the incubation period, the absorbance of each well at 450 nm was measured using a microplate reader, and cell viability was calculated according to the following formula:

[0153] Cell viability = (Average OD value of the drug or composition incubation group - Average OD value of the negative control group) / (Average OD value of the solvent control group - Average OD value of the negative control group) × 100%

[0154] like Figure 3 As shown, WX006 exhibits a good half-maximal inhibitory concentration (IC50) against the typical HCC cell line Hep3B. 50 The half-maximal inhibitory concentration (IC50) against Hep3B was 7.485 μM at 24 h and 3.535 μM at 48 h.

[0155] like Figure 4 As shown, WX006 exhibits a good half-maximal inhibitory concentration (IC50) against the typical HCC cell line Huh7. 50 The half-maximal inhibitory concentration (IC50) against Huh7 was 8.083 μM at 24 h and 3.751 μM at 48 h.

[0156] Drug safety testing

[0157] Primary liver cells from C57 mice were extracted using a collagenase perfusion digestion method, with 10 6Cells were seeded at 1 / mL and cultured for 24 hours. After adhesion, cells were treated with complete culture medium containing / without the target compound for 24 / 48 hours. Cell viability was assessed using the CCK8 assay, and the drug inhibition rate was calculated based on cell viability. Each dose point was repeated three times. IC50 values ​​were calculated using Graphpad Prism 8.0.2. Figure 5 As shown, no significant toxicity was observed after treating normal mouse hepatocytes with drug concentrations up to 10-30 times the IC50. This phenomenon indicates that the target compound has high tumor selectivity, or that primary hepatocytes can avoid the potential toxicity caused by high concentrations of the compound through some mechanism. This data suggests that the target compound has good efficacy as an anti-tumor drug and has a good therapeutic window.

[0158] Scratch test

[0159] Add 1 mL (2 × 10⁻⁶) to each well of a 12-well cell culture plate. 6 Cells were suspended in a cell / mL suspension and cultured until the cells reached over 90% confluence in the plate. Using a 200μL pipette tip perpendicular to the bottom of the plate, a straight line was drawn using a ruler and pipette tip. The original growth medium was then discarded, and the cells were washed three times with PBS to remove dead cells. 2.5mL of serum-free medium or serum-free medium containing WX006 was then added to each well for culture. Under this environment, cell proliferation was not observed, and cell migration ability was assessed. Each group was configured with three replicates. Three locations were selected at the marked site in each well, and images were taken at the same locations at 0h, 24h, and 48h after the mark. ImageJ was used for subsequent analysis of cell migration ability. Figure 6 As shown, in the colony formation experiment, the ability to inhibit proliferation significantly weakened, the number of colonies decreased, and this effect was concentration-dependent. These results indicate that the target compound has excellent anti-proliferative activity against Huh7 and Hep3B liver cancer cells. Figure 7 As shown, treatment of Hep3B and Huh7 cells with 10 μM target compound for 24 h and 48 h effectively inhibited cell scratch healing, demonstrating that the target compound can effectively inhibit the in vitro migration ability of liver cancer cells.

[0160] Western blotting (Western protein immunoblotting)

[0161] (1) Preparation of protein samples

[0162] Cells in good growth condition and in the logarithmic growth phase were subjected to a 3×10⁻⁶ thiocyanate infusion. 6Dot the desired number of cells per well into a 100mm culture dish. Once the cell confluence reaches approximately 75%, discard the original culture medium and add culture medium containing the drug stock solution or the corresponding volume of DMSO. Continue culturing until the drug treatment time is reached. Discard the drug-containing and blank culture media, wash twice with pre-cooled PBS, and repeat the process for each dish. 6 Add RIPA lysis buffer (pre-added with protease inhibitors and phosphatase inhibitors) at a ratio of cells / 100 μL. Incubate on ice for 5 min for lysis. Collect cells quickly with a pre-chilled cell scraper and transfer them to pre-chilled 1.5 mL centrifuge tubes. Centrifuge at 4 °C (12000 r / min, 15 min) and collect the supernatant as a backup sample for protein extraction. Store temporarily at -20 °C and immediately perform BCA quantification.

[0163] The protein concentration in each tube was determined according to the BCA kit instructions. After normalization, RIPA lysis buffer was added to the calibrated concentration to ensure that the concentrations in all sample tubes were consistent. Then, 5× Loading Buffer was added at a ratio of 4:1 and the mixture was quickly mixed using vortexing and pipette. The samples were then boiled in a 95°C metal bath for 5 minutes. After cooling and aliquoting, the aliquoted protein samples were stored at -80°C for later use.

[0164] (2) Electrophoresis

[0165] After installing the electrophoresis apparatus, add the sample and marker to the corresponding wells and record the data. Place the installed electrophoresis clamp into the electrophoresis tank and add 1× electrophoresis buffer to the 2-Gel mark. Select the constant voltage mode of 80V on the power output panel for electrophoresis. When the bromophenol blue front reaches the interface between the stacking gel and the separating gel, switch the electrophoresis voltage to a constant voltage of 120V. Stop electrophoresis when the bromophenol blue front is close to the bottom of the glass plate.

[0166] (3) Transfer membrane

[0167] Prepare filter paper, transfer clamps, and transfer sponges in advance and place them in pre-chilled 1× transfer buffer. Cut a PVDF membrane with an appropriate pore size of 0.22 μm or 0.45 μm according to the location of the protein to be transferred, and activate it in methanol for 5 min. After electrophoresis, cut the corresponding gel region according to the desired protein molecular weight and transfer the gel to filter paper on one side of the transfer clamp. Rinse the activated PVDF membrane repeatedly in transfer buffer until residual methanol is removed. Cover the gel surface with the equilibrated PVDF membrane, remove air bubbles by exposing it to light, and then stack the filter paper and sponge on the PVDF membrane. Clamp the transfer clamp and place the membrane in the transfer tank. Add pre-chilled 1× transfer buffer to the mark. Set the transfer conditions to a constant current of 320 mA and a time of 30 min for the target protein at its maximum molecular weight. Place the transfer system in an ice bath to ensure protein stability and transfer success rate.

[0168] (4) Closed

[0169] After the transfer was completed, the PVDF membrane was rinsed in 1×TBST for 5 min, the TBST was discarded, and blocking buffer (5% BSA for phosphorylated proteins and 5% skim milk powder for non-phosphorylated proteins) was added. The membrane was then placed on a shaker and incubated at 4°C for 16 hours to ensure complete blocking.

[0170] (5) Incubation of primary antibody

[0171] After the sealing process is completed, the strips are cut and placed on an acrylic plate. The prepared primary antibody working solution is added and the plate is incubated at 4°C for 16 hours in a humid environment.

[0172] (6) Washing the film

[0173] After incubation with the primary antibody, discard the working solution of the primary antibody, wash each band with 5 mL of TBST, and wash on a shaker at 100 r / min for 10 min, repeating 3 times.

[0174] (7) Incubation of secondary antibodies

[0175] Discard the TBST, place the corresponding band on an acrylic plate, add the prepared secondary antibody working solution, and incubate at room temperature in a humid environment for 2.5 hours.

[0176] (8) Washing the film

[0177] After the secondary antibody incubation is complete, discard the working solution of the secondary antibody, wash the band with 5 mL TBST, and wash on a shaker at 100 r / min for 10 min. Repeat 3 times.

[0178] (9) Development

[0179] The ECL luminescent solution was mixed evenly in a 1:1 ratio and dropped onto the PVDF film, then exposed using the ChemiDoc gel imaging system.

[0180] Experimental results are as follows Figure 8 As shown, the compound's parent nucleus caused downregulation of Cyclin D1, Cyclin E1, and Cyclin A2 in Hep3B cells, and Rb phosphorylation also decreased at 6h, 12h, and 24h, consistent with the cell cycle arrest results. This suggests that treatment of Hep3B cells with this compound can prevent cells from entering the S phase normally, causing G1 phase arrest by reducing cyclin expression. In Huh7 cells treated with this compound, Cyclin D1 and Cyclin E1 levels were significantly reduced at 6h, 12h, and 24h, while Cyclin A2 showed no significant change. However, Rb phosphorylation also decreased, indicating that this compound also caused normal cell cycle arrest in Huh7 cells.

[0181] Example 31

[0182] Two HCC tumor models were used to verify the tumor-suppressive effect and safety of WX006 in animals.

[0183] The specific steps are as follows:

[0184] Establishment of H22 hepatocellular carcinoma ascites tumor model in mice

[0185] H22 hepatocellular carcinoma ascites cells from the sixth generation and later were treated with physiological saline at a concentration of 1×10⁻⁶. 6 The cells were resuspended at a concentration of 1 cell / 50 μL. Using a sterile 1 mL syringe, the cell suspension was drawn up and injected subcutaneously into the right axilla of mice using the standard subcutaneous injection method. The injection volume was 50 μL. After needle removal, the mouse's tail was lifted, and a distinct raised area approximately 3 mm in diameter was observed in the right axilla, indicating successful model establishment. Throughout the implantation process, care was taken to avoid damaging the mouse's blood vessels and skin, ensuring no bleeding or interference with the mouse's physiological functions.

[0186] The longest length of the tumor is measured using calipers and recorded as L. Then, the width of the tumor, perpendicular to the length, is measured and recorded as W. The tumor volume (V) is calculated using the following formula:

[0187] Tumor volume (V) = L × W 2 ×0.5

[0188] One week after tumor implantation, the size of the axillary tumors in C57BL / 6 mice was assessed. After excluding outliers, mice were grouped according to tumor volume and body weight to ensure similar average tumor volume and body weight in each group, with 5 mice in each group. Tumor volume and body weight were measured daily for seven days. The experimental results are as follows: Figure 9 , Figure 10 As shown, mice underwent axillary tumor implantation on day 0, were grouped according to the initial tumor volume, and orally administered the target compound. On day 8, mice were sacrificed, and their internal organs and tumors were harvested for observation and analysis (n=5, mean±SEM). The experimental results indicate that the target compound effectively inhibited the subcutaneous growth of H22 hepatocellular carcinoma ascites tumors in C57 mice in a dose-dependent manner. Furthermore, the target compound did not significantly inhibit mouse body weight, demonstrating high safety.

[0189] During this period, mice were treated by gavage according to their group assignments, receiving 0.1 ml per 10 grams of body weight. At the end of the seven-day experimental period, all mice were euthanized, and tumors were removed for further analysis. The methods for observing and analyzing viscera and tumors were as follows: the visceral index (a) was calculated as visceral weight / mouse body weight (n=5, mean±SEM). Simultaneously, hematoxylin and eosin (H&E) staining was used to observe the pathological damage of the mouse viscera. Paraffin-embedded pathological sections of the left kidney, spleen, heart, and duodenum were prepared and stained with H&E. The microscopic pathological structures of different groups were observed and compared under a microscope. The results of the microscopic staining experiment are shown below. Figure 14 As shown, in kidney sections, the renal corpuscles in both the control and treatment groups remained morphologically intact, and the glomerular structures were clearly visible. In spleen sections, the red and white pulp structures in both the control and treatment groups were clearly visible. In heart sections, the myocardial fibers in both the control and treatment groups were densely arranged, with clear cross-sections; no intracardiac hemorrhage was observed. In small intestine sections, the small intestinal villi in both the control and treatment groups were clear, rounded, and tightly arranged. The interpretation of the microscopic pathological structure of mouse tissues showed that even at high doses, the target compound had almost no toxic effects on mouse organs, indicating high safety. The visceral index experiment results are as follows: Figure 11 , Figure 12 , Figure 13 As shown, the spleens of dissected mice were weighed and visceral indices were calculated. The results showed that the target compound had no significant effect on the weight of the spleen, liver, and kidneys of the mice, and no visceral toxicity was observed.

[0190] Establishment and in vivo fluorescence imaging of a mouse Hepa1-6-Luc orthotopic hepatocellular carcinoma model

[0191] Hepa1-6 cells were cultured normally in vitro and subjected to stress selection using 5 μg / ml puromycin. When the cells could grow normally to 80% confluence in 5 μg / ml puromycin, they could be used for in situ tumor implantation experiments.

[0192] Hepa1-6-luc cells adherent to the culture vessel were digested with 2.5% trypsin, resuspended in sterile saline, and the cell density was adjusted to 2 × 10⁻⁶ cells / year. 5 Cell / μL, keep on ice for later use.

[0193] Six- to seven-week-old male C57BL6 mice were selected and placed in a warm, clean environment. The oxygen flow rate was set to 5 L / min, and the induction anesthesia concentration was 2.5% (volume ratio). Mice were anesthetized approximately 15 seconds after being placed in the induction chamber. The exhaust gas flow rate was 35 L / min. After the righting reflex disappeared, a 3-4 mm incision was made 1 mm below the xiphoid process on the mouse's abdomen using sterile surgical instruments. The liver was exposed using the cotton swab compression method, and 2 × 10⁻⁶ mm x ... 6 One Hepa1-6-Luc cell was injected at a constant rate into the left lateral lobe of the mouse liver. Sterile gelatin sponge was used for hemostasis during the injection. Throughout the tumor implantation process, the mouse was anesthetized by inhaling 2% isoflurane via a face mask. After injection, the incision was quickly sutured with sterile needles and instruments. During the procedure, the mask anesthesia concentration was maintained at 2% (volume ratio), covering the entire process from implantation to wound closure.

[0194] Mice were placed on clean corncob bedding and provided with heating pads to maintain body temperature until they regained consciousness. The day of surgery was designated as day 0. On postoperative day 3, the tumors in the mice were quantitatively analyzed using the IVIS Spectrum (Perkin Elmer) in vivo imaging system, and the average values ​​were taken from each group. Drug administration was performed via gavage from day 4 to day 8 postoperatively. On postoperative day 9, a pre-prepared and sterile filtered 15 mg / mL fluorescein potassium solution was administered intraperitoneally at a dose of 100 μL per 10 g of mouse body weight. Ten minutes later, the tumors in the mice were again quantitatively assessed using the in vivo imaging system. The imaging system took images every 5, 10, 20, and 30 seconds. Images were considered stable when fluorescence intensity showed a linear relationship with imaging time. Quantitative analysis of fluorescent regions (ROIs) was performed using the accompanying Living Image 4.5 software.

[0195] Experimental results are as follows Figure 15 , Figure 16 , Figure 17 As shown, after axillary tumor implantation in mice on day 0, the mice were grouped according to the initial average tumor volume based on fluorescence quantitative quantification, and then administered the target compound orally (40 mg / kg). Quantification was performed again on day 6 after administration. After the oral administration cycle ended, the mice's body weight did not change significantly, and the tumor volume decreased significantly.

[0196] After the drug administration cycle, tumor-bearing mice were sacrificed, and their internal organs and tumors were harvested for observation and analysis. Ki67 immunohistochemical staining was used to observe the proliferation of in situ tumors in the mice (n = 5 mean ± SEM, *P < 0.05). Experimental results are as follows: Figure 18 As shown, oral administration of the target compound can reduce the expression of Ki67 in mouse orthotopic liver cancer.

[0197] Simultaneously, sorafenib, a widely used drug for treating liver cancer, was introduced for parallel comparison. Mice underwent axillary tumor implantation on day 0, and were grouped according to the initial average tumor volume based on quantitative PCR. Both mice were then orally administered the target compound and an equivalent dose of sorafenib (50 mg / kg) for 5 days. The experimental results are as follows: Figure 19 , Figure 20 , Figure 21 As shown, oral administration of the target compound reduced the size of in situ liver tumors in tumor-bearing mice, and was more effective than the same dose of sorafenib, without causing any weight loss in the mice.

[0198] As described above, although the invention has been shown and described with reference to specific preferred embodiments, it should not be construed as limiting the invention itself. Various changes in form and detail may be made without departing from the spirit and scope of the invention as defined in the appended claims.

Claims

1. An antitumor compound, characterized in that, The antitumor compound has the structure shown in general formula I: Among them, R1, R2, R3, and R4 are selected. The value of n is selected from one of the following: -H, -F, -Cl, -Br, -I, -CF3, -CCl3, -NO2, -COCH3, -OCOCH3, -CH3, -OCH3, -OC2H5, -NH2, -NHSO2 CH3, -SO2 NH2, NHCOCH3, where n is an integer selected from 1 to 5.

2. The compound according to claim 1, characterized in that: The antitumor compound is selected from the following compounds:

3. The compound according to claim 1 or 2, characterized in that: It also includes pharmaceutically acceptable salts, including acid addition salts formed by compounds of formula I with the following acids: hydrochloric acid, hydrobromic acid, sulfuric acid, phosphoric acid, methanesulfonic acid, benzenesulfonic acid, p-toluenesulfonic acid, naphthalenesulfonic acid, citric acid, tartaric acid, lactic acid, pyruvic acid, acetic acid, maleic acid or succinic acid, fumaric acid, salicylic acid, phenylacetic acid, and mandelic acid; and also includes acid salts formed by compounds of formula I with inorganic bases.

4. The compound according to claim 3, characterized in that: Pharmaceutically acceptable salts also include basic metal cation salts, alkaline earth metal cation salts, and ammonium cation salts.

5. A method for preparing the compound according to claim 1, characterized in that... Includes the following steps: (1) Dissolve 4,5-dibromo-1,2-dimethoxybenzene in dichloromethane, slowly add BBr3 dropwise under argon protection at -78°C, continue stirring and then move to room temperature; after the TLC monitoring shows that the raw material has basically reacted, add water to quench the residual BBr3, extract with dichloromethane 3 times, wash the organic layer 2 times with saturated sodium chloride solution, combine the organic layers, dry with anhydrous sodium sulfate, concentrate under reduced pressure to obtain white solid 4,5-dibromo-1,2-benzenediol; (2) Dissolve 4,5-dibromo-1,2-diphenol in acetone, add anhydrous potassium carbonate in batches under ice bath conditions, stir under mechanical stirrer, slowly add 1-chloromethoxy-2-methoxyethane to the reaction system, and react at room temperature for 1 h. After the reaction of the raw materials is completed by TLC monitoring, filter to remove anhydrous potassium carbonate from the system, concentrate under reduced pressure to remove acetone from the system, and after standing, the product solidifies from a reddish-brown oily liquid into pinkish white crystals. (3) Mix the white crystals obtained in step (2), 1,3-bis(diphenylphosphine)propane, and palladium acetate, and disperse them evenly in 1,4-dioxane. Add triethylamine and ethyl acrylate in sequence. Under argon protection, heat the reaction at 120°C and monitor the reaction until it is complete. Cool the system to room temperature, filter to remove insoluble matter, add ethyl acetate to dilute the filtrate, wash the organic phase with saturated sodium chloride solution, saturated sodium bicarbonate solution, and saturated ammonium chloride solution in sequence, dry the obtained organic phase with anhydrous sodium sulfate and concentrate under reduced pressure. The crude product is a reddish-brown oily liquid. (4) Dissolve the oily liquid obtained in step (3) in ethanol, add potassium hydroxide and stir thoroughly. After the reaction is complete by TLC monitoring, dilute the reaction solution with water, adjust the pH to 2 with hydrochloric acid under ice bath, extract with ethyl acetate three times, extract the organic phase with saturated sodium bicarbonate solution four times, convert the product into carboxylate and back-extract it into the aqueous phase, combine the aqueous phases, adjust the pH to 2 under ice bath, extract with ethyl acetate three times again, combine the organic phases, wash with saturated sodium chloride solution three times, dry the organic phase with anhydrous sodium sulfate, concentrate under reduced pressure, and obtain a brownish-gray solid as compound A. (5) Compound A and the substituted aromatic alcohol were dissolved in dichloromethane, and condensing agents 1-ethyl-3-(3-dimethylaminopropylamine)carbodiimide and 4-dimethylaminopyridine were added. A nitrogen protection device was added, and the mixture was stirred at room temperature. After the reaction of the raw materials was completed by TLC monitoring, the dichloromethane was evaporated to dryness, water was added for dilution, and the mixture was extracted three times with ethyl acetate. The organic phases were combined and washed three times with saturated sodium chloride solution. The organic phases were dried with anhydrous sodium sulfate and concentrated under reduced pressure. The crude product was purified by silica gel column chromatography to obtain a brown oily substance. (6) Dissolve the compound obtained in step (5) in methanol, then add 4N HCl, reflux at 70°C for 2 hours. After the reaction of the raw materials is completed by TLC monitoring, directly evaporate the reaction solution to obtain a light yellow solid. Extract with ethyl acetate, wash the organic layer twice with saturated sodium chloride solution, combine the organic layers and dry with anhydrous sodium sulfate, concentrate under reduced pressure, and precipitate the crude product by silica gel column chromatography to obtain a light yellow solid, which is the compound with the structure described in general formula I.

6. A method for preparing the compound according to claim 2, characterized in that... According to the preparation method of claim 5, the substituted aromatic alcohol in step (5) is phenylethanol.

7. Use of the compound according to any one of claims 1-4 in the preparation of a medicament for the prevention and / or treatment of tumors.

8. The use according to claim 7, characterized in that: The tumors include liver cancer, lung cancer, melanoma, colon cancer, diffuse large B-cell lymphoma, gastric cancer, glioma, and skin cancer.

9. A pharmaceutical composition, characterized in that, It comprises the compound as described in claim 1 and a pharmaceutically acceptable carrier or excipient.

10. The pharmaceutical composition according to claim 8, characterized in that, The pharmaceutical composition is a regular tablet or capsule, a sustained-release tablet or capsule, a controlled-release tablet or capsule, a granule, a powder, a syrup, an oral liquid, or an injection.

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