Napabucasin derivative with double-target antitumor activity as well as preparation method and application of Napabucasin derivative

Abstract

The invention relates to a Napabucasin derivative with double-target antitumor activity as well as a preparation method and application of the Napabucasin derivative. According to the invention, an STAT3 inhibitor Napabucasin is used as a parent nucleus, active structures such as zinc ion chelating group hydroxamic acid, o-phenylenediamine and alkane hydrazide of an HDAC inhibitor are introduced at the tail end of the STAT3 inhibitor Napabucasin through different Linkers, a series of Napabucasin derivative inhibitors with double-target antitumor activity are successfully obtained, the structural general formula of the Napabucasin derivative inhibitors is shown as (I), and pharmacological experiments show that the Napabucasin derivative inhibitors can be used for preparing antitumor drugs. The compound has high in-vitro anti-tumor activity, so that the compound can be used as an anti-tumor medicine. .

Description

Technical Field

[0001] This invention relates to Napabucasin derivatives with dual-target antitumor activity, their preparation methods, and applications, belonging to the field of pharmaceutical technology. Background Technology

[0002] Cancer has become one of the most serious threats to human health and life. Clinically, cancer is mainly treated using methods including single-target therapy and combination therapy with multiple drugs. However, these therapies have serious drawbacks such as significant side effects and poor patient compliance. Multi-target drugs, on the other hand, can act on multiple targets in the disease network simultaneously, producing a synergistic effect on each target, making the total effect greater than the sum of the individual effects, thus achieving optimal therapeutic efficacy. Furthermore, multi-target drugs have relatively simple pharmacokinetic properties and can overcome adverse reactions caused by drug-drug interactions. Therefore, multi-target drugs have become one of the important directions in the development of next-generation anti-tumor drugs.

[0003] Epigenetics, in contrast to genetics, primarily encompasses changes in gene expression levels not caused by alterations in gene sequence, such as chromatin conformational changes and DNA methylation. Cancer development is often closely related to epigenetic dysregulation. Studies have found that abnormal regulation of histone acetylation and deacetylation in the epigenetic system leads to changes in chromatin remodeling and spatial conformation, affecting the expression of normal genes and causing abnormalities in tumor suppressor genes or proteins related to cell cycle regulation, thereby promoting tumor development and metastasis. Therefore, the regulation of epigenetic targets has become a new direction in anti-tumor drug development, especially the study of epigenetic targets such as histone deacetylases (HDACs), which has become a current research hotspot.

[0004] Histone acetylation and histone deacetylation are two important regulatory mechanisms for gene expression and chromosome structure alterations. They play crucial roles in biological processes such as cellular transcription, translation, apoptosis, and energy metabolism. These two regulatory mechanisms are controlled by histone acetylases (HAT) and histone deacetylases (HDACs), respectively. HDACs hydrolyze and remove the N-terminal acetyl group from the lysine (Lys) side chain of histones, thereby making nucleosomes more compact and inhibiting gene transcription. They also regulate various cellular biological functions and processes, including gene expression, cell proliferation, differentiation, and apoptosis. Tumorigenesis and development are closely related to abnormal HDAC expression.

[0005] HDAC inhibitors (HDACi) have been shown to be effective in various biological processes, including inducing cell differentiation, inhibiting growth, promoting apoptosis, enhancing chemosensitivity, and inhibiting angiogenesis. Classic zinc-dependent HDAC inhibitors consist of three main parts: a hydrophobic group (Cap group) that recognizes the enzyme, a zinc-binding group (ZBG), and a linker connecting the two. The HDAC protein surface contains a hydrophobic region capable of binding to the Cap group. Introducing other antitumor active groups into the Cap terminus of HDAC inhibitors to design multi-target inhibitors helps enhance the binding interaction between the HDAC protein and the inhibitor, thereby improving the compound's antitumor activity. Several multi-target HDAC inhibitors are currently in clinical or preclinical research.

[0006] STAT3 (Statistical Inducer and Activator of Transcription 3) is a class of cellular transcription factors responsible for the transduction of extracellular cytokine and growth factor signals and the transcriptional activation of genes. The STAT3 signaling cascade is triggered by upstream kinase signals and undergoes phosphorylation, homodimerization, translocation into the nucleus, and binding to DNA, participating in a series of important physiological processes such as cell growth, proliferation, differentiation, and apoptosis. Under normal physiological conditions, STAT3 activation is rapid and transient. However, in various malignant tumors such as colorectal cancer, lung cancer, breast cancer, and kidney cancer, STAT3 is often persistently activated and expressed at high levels, playing a role in regulating cell proliferation, angiogenesis, metastasis, and anti-apoptosis, and is associated with tumor development and poor prognosis. Therefore, targeting STAT3 protein has great potential as a therapeutic strategy for tumors and related diseases.

[0007] Napabucasin (BBI-608) is a small molecule STAT3 inhibitor. Phase III clinical trials have shown its ability to inhibit STAT3-driven gene transcription and cancer stem cell characteristics, making it suitable for treating metastatic colorectal cancer, pancreatic cancer, and gastric cancer. In June and November 2016, Napabucasin was approved by the FDA as an orphan drug for the treatment of gastric or esophageal junction tumors and pancreatic cancer, respectively. Studies have shown that HDACi can inhibit the growth of hepatocellular carcinoma cells, malignant mast cells, and lung cancer cells by promoting STAT3 activation. Therefore, HDACi-based STAT3-positive cancer cell targeting may be an effective alternative anti-tumor therapy strategy. HDACi indirectly acts by enhancing BRD4 activation, thereby activating the leukemia inhibitory factor receptor (LIFR). Subsequently, LIFR can activate the downstream JAK1-STAT3 pathway. Ultimately, activated STAT3 can regulate target genes related to cell growth and metabolism, inhibit apoptosis, and accelerate cancer cell proliferation in response to hypoxia. Developed by Boston Biomedical (BBI), Napabucasin is one of the few STAT3 inhibitors to have entered Phase III clinical trials. Napabucasin has the potential to inhibit cancer cell metastasis and prevent cancer recurrence in various cancer types. Currently, the drug has not been approved for marketing in China and must be imported; furthermore, it has a single target. Summary of the Invention

[0008] In view of the shortcomings of the prior art, the first objective of this invention is to provide a class of Napabucasin derivatives with dual-target antitumor activity and pharmaceutically acceptable salts thereof.

[0009] A second objective of this invention is to provide a method for preparing the aforementioned Napabucasin derivative.

[0010] A third objective of this invention is to provide the application of the above-mentioned Napabucasin derivatives in the preparation of antitumor drugs.

[0011] This invention uses the STAT3 inhibitor Napabucasin as the core and introduces active structures such as isohydroxamic acid, o-phenylenediamine and alkane hydrazide, zinc ion chelating groups of HDAC inhibitors, through different linkers at its end, successfully obtaining a series of Napabucasin derivative inhibitors with dual-target anti-tumor activity.

[0012] To achieve the above objectives, the technical solution adopted by the present invention is as follows:

[0013] A class of Napabucasin derivatives or pharmaceutically acceptable salts thereof with dual-target antitumor activity, characterized in that the general structural formula of the Napabucasin derivative is as follows:

[0014]

[0015] in:

[0016] A is selected from hydroxyl group, ;

[0017] Where R1 is hydrogen, halogen, heterocyclic, R2 represents hydrogen or halogen.

[0018] The heterocycle is pyrrole, pyrazolyl, imidazolyl, furanyl, thiophene, oxazolyl, isoxazolyl, thiazolyl, isothiazolyl, pyridinyl, pyrimidinyl, pyrazinyl, or pyridazinyl.

[0019] R4 is selected from hydrogen, methyl, alkynyl, halogen, methoxy, trifluoromethyl, and cyano.

[0020] Where X is selected from n is an integer selected from 1 to 10.

[0021] R3 is selected from hydroxyl group, n is an integer selected from 1 to 6.

[0022] According to the present invention, the pharmaceutically acceptable salt is its organic acid salt or inorganic acid salt.

[0023] According to the present invention, the inorganic acid is hydrochloric acid, sulfuric acid, phosphoric acid, diphosphoric acid, hydrobromic acid or nitric acid; the organic acid is acetic acid, maleic acid, fumaric acid, tartaric acid, succinic acid, lactic acid, p-toluenesulfonic acid, salicylic acid, oxalic acid, tannic acid, citric acid, trifluoroacetic acid, malic acid or benzenesulfonate.

[0024] According to the present invention, more preferably, the Napabucasin derivative is any one of the following:

[0025] Compound XC-1: N-(2-aminophenyl)-4,9-dioxo-4,9-dihydronaphthoxy[2,3-b]furan-2-carboxamide,

[0026] Compound XC-2: N-(3-((2-aminophenyl)amino)-3-oxopropyl)-4,9-dioxo-4,9-dihydronaphtho[2,3-b]furan-2-carboxamide,

[0027] Compound XC-3: N-(2-amino-5-fluorophenyl)-4,9-dioxo-4,9-dihydronaphtho[2,3-b]furan-2-carboxamide,

[0028] Compound XC-4: N-(2-amino-4,5-difluorophenyl)-4,9-dioxo-4,9-dihydronaphtho[2,3-b]furan-2-carboxamide,

[0029] Compound XC-5: N-(4-amino-[1,1'-biphenyl]-3-yl)-4,9-dioxo-4,9-dihydronaphtho[2,3-b]furan-2-carboxamide,

[0030] Compound XC-6: N-(4-amino-4'-fluoro-[1,1'-biphenyl]-3-yl)-4,9-dioxo-4,9-dihydronaphtho[2,3-b]furan-2-carboxamide,

[0031] Compound XC-7: N-(4-amino-4'-methyl-[1,1'-biphenyl]-3-yl)-4,9-dioxo-4,9-dihydronaphtho[2,3-b]furan-2-carboxamide,

[0032] Compound XC-8: N-(4-amino-4'-chloro-biphenyl-3-yl)-4,9-dioxo-4,9-dihydronaphtho[2,3-b]furan-2-carboxamide,

[0033] Compound XC-9: N-(4-amino-4'-methoxy-biphenyl-3-yl)-4,9-dioxo-4,9-dihydronaphtho[2,3-b]furan-2-carboxamide,

[0034] Compound XC-10: N-(2-amino-5-(pyridin-4-yl)phenyl)-4,9-dioxo-4,9-dihydronaphtho[2,3-b]furan-2-carboxamide,

[0035] Compound XC-11: N-(2-amino-5-(pyridin-3-yl)phenyl)-4,9-dioxo-4,9-dihydronaphtho[2,3-b]furan-2-carboxamide,

[0036] Compound XC-12: N-(2-amino-5-(furan-2-yl)phenyl)-4,9-dioxo-4,9-dihydronaphtho[2,3-b]furan-2-carboxamide,

[0037] Compound XC-13: N-(2-amino-5-(thiophen-2-yl)phenyl)-4,9-dioxo-4,9-dihydronaphtho[2,3-b]furan-2-carboxamide,

[0038] Compound XC-14: N-(2-amino-5-(thiophen-3-yl)phenyl)-4,9-dioxo-4,9-dihydronaphtho[2,3-b]furan-2-carboxamide,

[0039] Compound XC-15: N-(4-amino-4'-(trifluoromethyl)-biphenyl-3-yl)-4,9-dioxo-4,9-dihydronaphtho[2,3-b]furan-2-carboxamide,

[0040] Compound XC-16: N-(4-amino-4'-cyano-biphenyl-3-yl)-4,9-dioxo-4,9-dihydronaphtho[2,3-b]furan-2-carboxamide,

[0041] Compound XC-17: N-hydroxy-4,9-dioxo-4,9-dihydronaphtho[2,3-b]furan-2-carboxamide,

[0042] Compound XC-18: N-(4-(hydroxycarbamoyl)phenyl)-4,9-dioxo-4,9-dihydronaphtho[2,3-b]furan-2-carboxamide,

[0043] Compound XC-19: N-(3-(hydroxyamino)-3-oxopropyl)-4,9-dioxo-4,9-dihydronaphtho[2,3-b]furan-2-carboxamide,

[0044] Compound XC-20: N-(5-(hydroxyamino)-5-oxopentyl)-4,9-dioxo-4,9-dihydronaphtho[2,3-b]furan-2-carboxamide,

[0045] Compound XC-21: N-(6-(hydroxyamino)-6-oxohexyl)-4,9-dioxo-4,9-dihydronaphtho[2,3-b]furan-2-carboxamide,

[0046] Compound XC-22: N-(7-(hydroxyamino)-7-oxohepyl)-4,9-dioxo-4,9-dihydronaphtho[2,3-b]furan-2-carboxamide,

[0047] Compound XC-23: N-(8-(hydroxyamino)-8-oxooctyl)-4,9-dioxo-4,9-dihydronaphtho[2,3-b]furan-2-carboxamide,

[0048] Compound XC-24: N-(9-(hydroxyamino)-9-oxonyl)-4,9-dioxo-4,9-dihydronaphtho[2,3-b]furan-2-carboxamide,

[0049] Compound XC-25: N'-ethyl-4,9-dioxo-4,9-dihydronaphtho[2,3-b]furan-2-carboxylhydrazine,

[0050] Compound XC-26: 4,9-dioxo-N'-propyl-4,9-dihydronaphtho[2,3-b]furan-2-formylhydrazine,

[0051] Compound XC-27: N'-butyl-4,9-dioxo-4,9-dihydronaphtho[2,3-b]furan-2-formylhydrazine,

[0052] Compound XC-28: 4,9-dioxo-N'-pentyl-4,9-dihydronaphtho[2,3-b]furan-2-formylhydrazine,

[0053] Compound XC-29: N'-isopentyl-4,9-dioxo-4,9-dihydronaphtho[2,3-b]furan-2-carboxylhydrazine.

[0054] The preparation method of the above-mentioned Napabucasin derivatives.

[0055] Specifically:

[0056] According to a preferred embodiment of the present invention, the method for synthesizing compounds XC-1 to XC-16 includes the following steps:

[0057] Compound 5 is condensed with compound 14 to give compound 15. Compound 15 is de-tert-butylated under acidic conditions to give compound 16. Compound 16 is condensed with o-phenylenediamine 17 protected by a Boc group to give compound 18. Compound 18 is de-Bocated under acidic conditions to give the target compound XC-2. Alternatively, compound 5 is condensed with o-phenylenediamine 19a-o with different substituents to give compound 20a-o. Compound 20a-o is de-Bocated under acidic conditions to give the target compound XC-1 or XC-3~16.

[0058] The reaction route is as follows:

[0059] Reagents and conditions: (a) HBTU, DIPEA, DMF, room temperature, 4 h, yield 64-74%; (b) TFA, room temperature, 1 h, yield 93-95%.

[0060] According to a preferred embodiment of the present invention, the method for synthesizing compounds XC-17 to XC-24 includes the following steps:

[0061] Compound 5 is directly condensed with o-triphenylhydroxylamine 24 to give compound 26. Compound 26 is reacted with TFA to remove the protecting group to give the target compound XC-17; or, compound 5 is condensed with compound 21 to give compound 22. Compound 22 is de-tert-butylated under acidic conditions to give compound 23. Compound 23 is then condensed with o-triphenylhydroxylamine 24 to give compound 25. Compound 25 is reacted with TFA to remove the protecting group to give the target compound XC-18; or, compound 5 is condensed with alkane chains of different lengths to give compounds 28a-f. Compound 28a-f is de-tert-butylated under acidic conditions to give compounds 29a-f. Compound 29a-f is then condensed with o-triphenylhydroxylamine 24 to give compounds 30a-f. Compound 25 is reacted with TFA to remove the protecting group to give the target compounds XC-19~24.

[0062] The reaction route is as follows:

[0063]

[0064] Reagents and conditions: (a) HBTU, DIPEA, DMF, room temperature, 4 h, yield 64-74%; (b) TFA, room temperature, 1 h, yield 93-95%.

[0065] According to a preferred embodiment of the present invention, the method for synthesizing compounds XC-25 to XC-29 includes the following steps:

[0066] Compound 5 is condensed with Boc-protected hydrazides of different chain lengths 31a-d to generate compounds 32a-d. Compound 32a-d is de-tert-butylated under acidic conditions to give the target compounds XC-25~28. Alternatively, compound 5 is condensed with compound 33 to generate compound 34. Compound 34 is de-tert-butylated under acidic conditions to give the target compound XC-29.

[0067] The reaction route is as follows:

[0068]

[0069] Reagents and conditions: (a) HBTU, DIPEA, DMF, room temperature, 4 h, yield 64-74%; (b) TFA, room temperature, 1 h, yield 93-95%.

[0070] A third objective of this invention is to provide the use of the above-mentioned Napabucasin derivatives or pharmaceutically acceptable salts thereof in the preparation of antitumor drugs or drugs for differentiation and proliferation-related diseases.

[0071] According to a preferred embodiment of the present invention, the tumor is selected from colorectal cancer, breast cancer, lung cancer, liver cancer, and prostate cancer.

[0072] According to a preferred embodiment of the present invention, the medicinal salt is free of water of crystallization, or contains one or more molecules of water of crystallization.

[0073] The use of the above-mentioned Napabucasin derivatives or their pharmaceutically acceptable salts in the preparation of HDAC inhibitors, STAT3 inhibitors or HDAC / STAT3 dual-target inhibitors.

[0074] The present invention has the following advantages and beneficial effects:

[0075] 1. This invention uses the STAT3 inhibitor Napabucasin as the core and introduces active structures such as isohydroxamic acid, o-phenylenediamine, and alkane hydrazide, which are zinc ion chelating groups of HDAC inhibitors, into its terminal through different linkers. A series of Napabucasin derivatives with dual-target antitumor activity have been successfully obtained. These are small molecule anticancer drugs based on the dual targets of STAT3 and HDAC. Pharmacological experiments show that the Napabucasin derivatives of this invention have strong in vitro antitumor activity and can therefore be used as antitumor drugs.

[0076] 2. The compound of this invention exhibits strong inhibitory activity against histone deacetylase 1 and can effectively inhibit STAT3 phosphorylation. It also possesses strong in vitro antitumor activity, targeting tumors including colorectal cancer, breast cancer, leukemia, lung cancer, liver cancer, and prostate cancer. Attached Figure Description

[0077] Figure 1 Phosphorylation results of STAT3 by compounds XC-10 and XC-22. Detailed Implementation

[0078] To more clearly illustrate the present invention, the following description, in conjunction with preferred embodiments, further clarifies the invention. Those skilled in the art should understand that the specific descriptions below are illustrative rather than restrictive, and should not be construed as limiting the scope of protection of the present invention.

[0079] Experimental methods in the following examples, unless otherwise specified, are generally performed under standard conditions or as recommended by the manufacturer. Unless otherwise stated, all percentages mentioned in the examples are by weight.

[0080] Example 1: Preparation of compound XC-1:

[0081]

[0082] The reaction route is as follows:

[0083]

[0084] The specific preparation method and steps are as follows:

[0085] (1) Preparation of compound 20a:

[0086]

[0087] Compound 5 (30.00 mg, 0.12 mmol) and compound 19a (31.22 mg, 0.15 mmol) were dissolved in 2 mL of DMF, and then HATU (72.25 mg, 0.19 mmol) and DIPEA (24.56 mg, 0.19 mmol) were added. The mixture was stirred at room temperature for 4 h. The reaction was monitored by TLC, with PE:EA = 1:1 as the developing solvent. After the reaction was complete, the mixture was extracted three times with water and EA. The upper organic phases were combined, concentrated, mixed with silica gel, and purified by column chromatography (PE:EA = 85:15) to obtain compound 20a.

[0088] 1 H NMR (400 MHz, DMSO-d6) δ 10.15 (s, 1H), 8.89 (s, 1H), 8.19 – 8.13(m, 2H), 7.95 – 7.91 (m, 2H), 7.82 (s, 1H), 7.61 (dd, J = 8.1, 1.5 Hz, 1H), 7.51 (dd, J = 8.0, 1.6 Hz, 1H), 7.25 (m, 1H), 7.16 (m, 1H), 1.47 (s, 9H).

[0089] (2) Preparation of compound XC-1:

[0090]

[0091] Compound 20a was dissolved in 6 mL of DCM, and then 2 mL of TFA was added. The mixture was stirred at room temperature for 1 h. After the reaction was complete, the solvent was evaporated, and the mixture was extracted with H2O / DCM. The lower organic phases were combined and concentrated to obtain compound XC-1.

[0092] 1H NMR (400 MHz, DMSO-d6) δ 10.06 (s, 1H), 8.16 (m, J = 9.8, 6.5, 3.4Hz, 2H), 7.97 – 7.90 (m, 2H), 7.88 (s, 1H), 7.17 – 7.13 (m, 1H), 7.04 – 6.98 (m, 1H), 6.78 (dd, J = 8.1, 1.4 Hz, 1H), 6.63 – 6.58 (m, 1H), 5.07 (s, 2H).

[0093] Example 2

[0094] Preparation of compound XC-2:

[0095]

[0096] The reaction route is as follows:

[0097]

[0098] The specific preparation method and steps are as follows:

[0099] (1) Preparation of compound 15

[0100]

[0101] Compound 14 (36.59 mg, 0.25 mmol) and compound 5 (50.00 mg, 0.21 mmol) were dissolved in 2 mL of DMF, and then HATU (117.87 mg, 0.31 mmol) and DIPEA (40.07 mg, 0.31 mmol) were added. The mixture was stirred at room temperature for 4 h. The reaction was monitored by TLC, with PE:EA = 2:1 as the developing solvent. After the reaction was complete, the mixture was extracted three times with water and EA. The upper organic phases were combined, concentrated, and then mixed with silica gel for purification by column chromatography (PE:EA = 80:20) to obtain compound 15 as a yellow solid.

[0102] 1 H NMR (400 MHz, DMSO-d6) δ 9.00 (t, J = 5.7 Hz, 1H), 8.17 – 8.09 (m,2H), 7.94 – 7.88 (m, 2H), 7.62 (s, 1H), 3.48 (q, J = 6.6 Hz, 2H), 3.16 (d, J= 5.3 Hz, 2H), 1.40 (s, 9H).

[0103] (2) Preparation of compound 18

[0104]

[0105] Compound 15 (60.00 mg) was dissolved in 6 mL of DCM, and then 2 mL of TFA was added. The mixture was stirred at room temperature for 1 h until the reaction was complete. After the reaction was complete, the solvent was evaporated, and the TFA was removed. A small amount of DIPEA was added to neutralize the residual TFA. Then, the mixture was dissolved in 2 mL of DMF, and HATU (91.25 mg, 0.24 mmol) and DIPEA (31.02 mg, 0.24 mmol) were added. Compound 5 (33.30 mg, 0.16 mmol) was added. The mixture was stirred at room temperature for 4 h. The reaction was monitored by TLC using a PE:EA ratio of 1:2 as the developing solvent. After the reaction was complete, the mixture was extracted three times with water and EA. The upper organic phases were combined, concentrated, mixed with silica gel, and purified by column chromatography (PE:EA = 50:50) to obtain compound 18.

[0106] 1 H NMR (400 MHz, DMSO-d6) δ 9.46 (s, 1H), 9.09 (t, J = 5.6 Hz, 1H), 8.43 (s, 1H), 8.17 – 8.09 (m, 2H), 7.93 – 7.88 (m, 2H), 7.65 (s, 1H), 7.56(d, J = 8.0 Hz, 1H), 7.47 (dd, J = 7.8, 1.7 Hz, 1H), 7.09 (m, 2H), 3.59 (q, J= 6.6 Hz, 2H), 2.68 (s, 2H), 1.43 (s, 9H).

[0107] (3) Preparation of compound XC-2

[0108]

[0109] Compound 18 was dissolved in 6 mL of DCM, and then 2 mL of TFA was added. The mixture was stirred at room temperature for 1 h. After the reaction was complete, the solvent was evaporated, and the mixture was extracted with H2O / DCM. The lower organic phases were combined and concentrated to give compound XC-2 in 68% yield.

[0110] 1H NMR (400 MHz, DMSO-d6) δ 9.22 (s, 1H), 9.08 (t, J = 5.7 Hz, 1H), 8.13 (m, 2H), 7.93 – 7.88 (m, 2H), 7.66 (s, 1H), 7.15 (dd, J = 7.8, 1.5 Hz,1H), 6.90 (m, 1H), 6.71 (dd, J = 7.9, 1.5 Hz, 1H), 6.54 (m, 1H), 3.59 (q, J =6.6 Hz, 2H), 2.65 (t, J = 6.9 Hz, 2H).

[0111] Example 3

[0112] Preparation of compound XC-3: In Example 1, compound 9a was replaced with compound 9b in the first step, and the rest was the same as in Example 1. Compound XC-3 was prepared with a yield of 72%.

[0113] Example 4

[0114] Preparation of compound XC-4: In Example 1, compound 9a was replaced with compound 9c in the first step, and the rest was the same as in Example 1. Compound XC-4 was prepared with a yield of 75%.

[0115] Example 5

[0116] Preparation of compound XC-5: In Example 1, compound 9a was replaced with compound 9d in the first step, and the rest was the same as in Example 1. Compound XC-5 was prepared with a yield of 70%.

[0117] Example 6

[0118] Preparation of compound XC-6: In Example 1, compound 9a was replaced with compound 9e in the first step, and the rest was the same as in Example 1. Compound XC-6 was prepared with a yield of 65%.

[0119] Example 7

[0120] Preparation of compound XC-7: In Example 1, compound 9a was replaced with compound 9f in the first step, and the rest was the same as in Example 1. Compound XC-7 was prepared with a yield of 66%.

[0121] Example 8

[0122] Preparation of compound XC-8: In Example 1, compound 9a was replaced with compound 9g in the first step, and the rest was the same as in Example 1. Compound XC-8 was prepared with a yield of 78%.

[0123] Example 9

[0124] Preparation of compound XC-9: In Example 1, compound 9a was replaced with compound 9h in the first step, and the rest was the same as in Example 1. Compound XC-9 was prepared with a yield of 56%.

[0125] Example 10

[0126] Preparation of compound XC-10: In Example 1, compound 9a was replaced with compound 9i in the first step, and the rest was the same as in Example 1. Compound XC-10 was prepared with a yield of 68%.

[0127] Example 11

[0128] Preparation of compound XC-11: In Example 1, compound 9a was replaced with compound 9j in the first step, and the rest was the same as in Example 1. Compound XC-11 was prepared with a yield of 63%.

[0129] Example 12

[0130] Preparation of compound XC-12: In Example 1, compound 9a was replaced with compound 9k in the first step, and the rest was the same as in Example 1. Compound XC-12 was prepared with a yield of 63%.

[0131] Example 13

[0132] Preparation of compound XC-13: In Example 1, compound 9a was replaced with compound 9l in the first step, and the rest was the same as in Example 1. Compound XC-13 was prepared with a yield of 64%.

[0133] Example 14

[0134] Preparation of compound XC-14: In Example 1, compound 9a was replaced with compound 9m in the first step, and the rest was the same as in Example 1. Compound XC-14 was prepared with a yield of 71%.

[0135] Example 15

[0136] Preparation of compound XC-15: In Example 1, compound 9a was replaced with compound 9n in the first step, and the rest was the same as in Example 1. Compound XC-15 was prepared with a yield of 65%.

[0137] Example 16

[0138] Preparation of compound XC-16: In Example 1, compound 9a was replaced with compound 9o in the first step, and the rest was the same as in Example 1. Compound XC-16 was prepared with a yield of 77%.

[0139] Example 17

[0140] Preparation of compound XC-17

[0141]

[0142] (1) Preparation of compound 26

[0143]

[0144] Compound 5 (30.00 mg, 0.12 mmol) and compound 24 (44.05 mg, 0.16 mmol) were dissolved in 2 mL of DMF, and then HATU (72.25 mg, 0.19 mmol) and DIPEA (24.56 mg, 0.19 mmol) were added. The mixture was stirred at room temperature for 4 h. The reaction was monitored by TLC with PE:EA = 2:1 as the developing solvent. After the reaction was complete, the mixture was extracted three times with water and EA. The upper organic phases were combined, concentrated, and then purified by column chromatography (PE:EA = 70:30) to obtain compound 26.

[0145] 1 H NMR (400 MHz, DMSO-d6) δ 8.12 – 8.06 (m, 2H), 7.91 – 7.86 (m, 2H), 7.41 (s, 1H), 7.40 – 7.29 (m, 15H).

[0146] (2) Preparation of compound XC-17

[0147]

[0148] Compound 26 was dissolved in 6 mL of DCM, and then 2 mL of TFA was added. The mixture was stirred at room temperature for 0.5 h. After the reaction was complete, the solvent was evaporated, the mixture was slurried with methanol, and filtered to obtain a pale yellow solid compound XC-17 with a yield of 55%.

[0149] 1 H NMR (400 MHz, DMSO-d6) δ 11.76 (s, 1H), 9.53 (s, 1H), 8.12 (m, 2H), 7.94 – 7.88 (m, 2H), 7.55 (s, 1H).

[0150] Example 18

[0151] Preparation of compound XC-18

[0152]

[0153] (1) Preparation of compound 22

[0154]

[0155] Compound 21 (39.89 mg, 0.21 mmol) and compound 5 (50.00 mg, 0.21 mmol) were dissolved in 2 mL of DMF, and then HATU (117.87 mg, 0.31 mmol) and DIPEA (40.07 mg, 0.31 mmol) were added. The mixture was stirred at room temperature for 4 h. The reaction was monitored by TLC, with PE:EA = 3:1 as the developing solvent. After the reaction was complete, the mixture was slurried with methanol and filtered to obtain a pale yellow solid, compound 22.

[0156] (2) Preparation of compound 25

[0157]

[0158] Compound 22 (54.00 mg, 0.13 mmol) was dissolved in 6 mL of DCM, followed by the addition of 2 mL of TFA. The mixture was stirred at room temperature for 0.5 h until the reaction was complete. The solvent was then evaporated, and the TFA was removed by evaporation. A small amount of DIPEA was added to neutralize the residual TFA. The mixture was then dissolved in 2 mL of DMF, followed by the addition of HATU (76.05 mg, 0.20 mmol) and DIPEA (25.85 mg, 0.20 mmol), and compound 24 (35.79 mg, 0.13 mmol). The reaction was stirred at room temperature for 4 h. The reaction was monitored by TLC using a PE:EA ratio of 2:1 as the developing solvent. After the reaction was complete, the mixture was extracted three times with water and EA. The upper organic phases were combined, concentrated, and then purified by column chromatography (PE:EA = 70:30) to obtain compound 25 as a yellow solid.

[0159] (3) Preparation of compound XC-18

[0160]

[0161] Compound 25 was dissolved in 6 mL of DCM, and then 2 mL of TFA was added. The mixture was stirred at room temperature for 0.5 h. After the reaction was complete, the solvent was evaporated, the mixture was slurried with methanol, and filtered to obtain a pale yellow solid compound XC-18 with a yield of 66%.

[0162] 1 H NMR (400 MHz, DMSO-d6) δ 11.19 (s, 1H), 10.85 (s, 1H), 8.16 (m,2H), 7.96 (s, 1H), 7.95 – 7.91 (m, 2H), 7.86 (d, J = 8.8 Hz, 2H), 7.79 (d, J= 8.7 Hz, 2H).

[0163] Example 19

[0164] Preparation of compound XC-19

[0165]

[0166] (1) Preparation of compound 28a

[0167]

[0168] Compound 27a (36.59 mg, 0.25 mmol) and compound 5 (50.00 mg, 0.21 mmol) were dissolved in 2 mL of DMF, and then HATU (117.87 mg, 0.31 mmol) and DIPEA (40.07 mg, 0.31 mmol) were added. The mixture was stirred at room temperature for 4 h. The reaction was monitored by TLC with PE:EA = 2:1 as the developing solvent. After the reaction was complete, the mixture was extracted three times with water and EA. The upper organic phases were combined, concentrated, and then mixed with silica gel for purification by column chromatography (PE:EA = 80:20) to obtain compound 28a as a yellow solid.

[0169] 1 H NMR (400 MHz, DMSO-d6) δ 9.00 (t, J = 5.7 Hz, 1H), 8.17 – 8.09 (m,2H), 7.94 – 7.88 (m, 2H), 7.62 (s, 1H), 3.48 (q, J = 6.6 Hz, 2H), 3.16 (d, J= 5.3 Hz, 2H), 1.40 (s, 9H).

[0170] (2) Preparation of compound 30a

[0171]

[0172] Compound 28a (60.00 mg) was dissolved in 6 mL of DCM, and then 2 mL of TFA was added. The mixture was stirred at room temperature for 0.5 h until the reaction was complete. After the reaction was complete, the solvent was evaporated, and the TFA was removed by evaporation. A small amount of DIPEA was added to neutralize the residual TFA. Then, the mixture was dissolved in 2 mL of DMF, and HATU (91.25 mg, 0.24 mmol) and DIPEA (31.02 mg, 0.24 mmol) were added. Compound 24 (44.05 mg, 0.16 mmol) was added. The reaction was stirred at room temperature for 4 h. The reaction was monitored by TLC using PE:EA = 1:1 as the developing solvent. After the reaction was complete, the mixture was extracted three times with water and EA. The upper organic phases were combined, concentrated, and then mixed with silica gel for purification by column chromatography (PE:EA = 60:40) to obtain compound 30a as a pale yellow solid.

[0173] 1 H NMR (400 MHz, DMSO-d6) δ 10.42 (s, 1H), 8.78 (d, J = 6.4 Hz, 1H), 8.17 – 8.09 (m, 2H), 7.93 – 7.89 (m, 2H), 7.58 (s, 1H), 7.36 – 7.26 (m, 15H),3.21 – 3.14 (m, 2H), 2.11 (t, J = 7.2 Hz, 2H).

[0174] (3) Preparation of compound XC-19

[0175]

[0176] Compound 30a was dissolved in 6 mL of DCM, and then 2 mL of TFA was added. The mixture was stirred at room temperature for 0.5 h. After the reaction was complete, the solvent was evaporated, the mixture was slurried with methanol, and filtered to obtain a pale yellow solid compound XC-19 with a yield of 57%.

[0177] 1 H NMR (400 MHz, DMSO-d6) δ 10.49 (d, J = 1.7 Hz, 1H), 9.03 (t, J =5.7 Hz, 1H), 8.79 (d, J = 1.7 Hz, 1H), 8.13 (m, 2H), 7.94 – 7.88 (m, 2H), 7.63 (s, 1H), 3.47 (q, J = 6.7 Hz, 2H), 2.28 (t, J = 7.1 Hz, 2H).

[0178] Example 20

[0179] Preparation of compound XC-20: In Example 19, compound 27a was replaced with compound 27b in the first step, and the rest was the same as in Example 19. Compound XC-20 was prepared with a yield of 63%.

[0180] Example 21

[0181] Preparation of compound XC-21: In Example 19, compound 27a was replaced with compound 27c in the first step, and the rest was the same as in Example 19. Compound XC-21 was prepared with a yield of 66%.

[0182] Example 22

[0183] Preparation of compound XC-22: In Example 19, compound 27a was replaced with compound 27d in the first step, and the rest was the same as in Example 19. Compound XC-22 was prepared with a yield of 59%.

[0184] Example 23

[0185] Preparation of compound XC-23: In Example 19, compound 27a was replaced with compound 27e in the first step, and the rest was the same as in Example 19. Compound XC-23 was prepared with a yield of 58%.

[0186] Example 24

[0187] Preparation of compound XC-24: In Example 19, compound 27a was replaced with compound 27f in the first step, and the rest was the same as in Example 19. Compound XC-24 was prepared with a yield of 60%.

[0188] Example 25

[0189] Preparation of compound XC-25

[0190]

[0191] (1) Preparation of compound 32a

[0192]

[0193] Compound 5 (50.00 mg, 0.21 mmol) and compound 31a (33.64 mg, 0.21 mmol) were dissolved in 2 mL of DMF, and then HATU (121.68 mg, 0.32 mmol) and DIPEA (40.71 mg, 0.32 mmol) were added. The mixture was stirred at room temperature for 4 h. The reaction was monitored by TLC with PE:EA = 1:1 as the developing solvent. After the reaction was complete, the mixture was extracted three times with water and EA. The upper organic phases were combined, concentrated, and then mixed with silica gel for purification by column chromatography (PE:EA = 85:15) to obtain compound 32a.

[0194] 1 H NMR (400 MHz, DMSO-d6) δ 11.05 (s, 1H), 8.14 (m, 2H), 7.95 – 7.89(m, 2H), 7.74 (s, 1H), 3.47 (q, J = 7.1 Hz, 2H), 1.48 – 1.32 (m, 9H), 1.10(s, 3H).

[0195] (2) Preparation of compound XC-25

[0196]

[0197] Compound 32a was dissolved in 6 mL of DCM, and then 2 mL of TFA was added. The mixture was stirred at room temperature for 1 h. The reaction was monitored by TLC, with PE:EA = 1:1 as the developing solvent. After the reaction was complete, the solvent was evaporated, and the mixture was extracted with H2O / DCM. The lower organic phases were combined and concentrated to give compound XC-25 in 56% yield.

[0198] 1 H NMR (400 MHz, DMSO-d6) δ 10.46 (s, 1H), 8.12 (m, 2H), 7.94 – 7.86(m, 2H), 7.63 (s, 1H), 5.27 (s, 1H), 2.84 (q, J = 7.2 Hz, 2H), 1.04 (t, J =7.1 Hz, 3H).

[0199] Example 26

[0200] Preparation of compound XC-26: In Example 25, compound 31a was replaced with compound 31b in the first step, and the rest was the same as in Example 25. Compound XC-26 was prepared with a yield of 70%.

[0201] Example 27

[0202] Preparation of compound XC-27: In Example 25, compound 31a was replaced with compound 31c in the first step, and the rest was the same as in Example 25. Compound XC-27 was prepared with a yield of 69%.

[0203] Example 28

[0204] Preparation of compound XC-28: In Example 25, compound 31a was replaced with compound 31d in the first step, and the rest was the same as in Example 25. Compound XC-28 was prepared with a yield of 67%.

[0205] Example 29

[0206] Preparation of compound XC-29: In Example 25, compound 31a in the first step was replaced with compound 33, and the rest was the same as in Example 25. Compound XC-29 was prepared with a yield of 65%.

[0207] The structural formulas and NMR data of compounds XC-1 to XC-29 are shown in Table 1 below:

[0208] Table 1. Structural formulas and NMR data of the compounds

[0209]

[0210]

[0211]

[0212]

[0213] Experimental examples of target compound enzyme inhibitory activity and in vitro antitumor activity

[0214] 1. Inhibition test of target compound on HDAC1 enzyme activity

[0215] 1.1 Experimental Materials:

[0216] HDAC1 enzyme, buffer (137mM sodium chloride, 2.7mM potassium chloride, 1mM magnesium chloride, 0.1 mg / mL BSA, 25mM Tris-HCl at pH=8), HDAC substrate 3, trypsin, 96-well black plate.

[0217] 1.2 Experimental Methods:

[0218] (1) Equilibrate the 96-well black plate to room temperature;

[0219] (2) Dilute the test compound with a buffer containing 10% DMSO. The concentrations of the compounds are 100 μM, 30 μM, 10 μM, 3 μM, 1 μM, 0.3 μM, 0.1 μM, 0.03 μM, 0.01 μM, and 0.003 μM, respectively.

[0220] (3) Add 11 μL of HDAC1 to 400 μL of buffer solution and shake well;

[0221] (4) Add 35 μL of the prepared HDAC1 enzyme buffer to wells 2-11 of the 96-well plate, and add 5 μL of the diluted compounds of different concentrations to the corresponding reaction wells in sequence. For the negative control (first well) and the blank control (eleventh well), add 40 μL and 5 μL of assay buffer, respectively.

[0222] (5) Add 5 μL of 100 μM HDAC substrate and 5 μL of 0.5 mg / mL trypsin to all reaction wells, incubate at 37°C for 30 minutes and then take the readings.

[0223] (6) Calculate the inhibition rate according to the formula: Inhibition rate = (100% active wells - sample wells) / 100% active wells * 100. Fit the enzyme activity versus compound concentration curve in GraphPad software to determine the IC50 of the compound. 50 value;

[0224] Experimental results showed that these compounds all exhibited good HDAC1 inhibitory activity, with some compounds showing superior activity compared to the positive control drug SAHA (vorinostat) (IC50). 50 HDAC1 inhibitory activity (0.041 μM) was measured.

[0225] Experimental results showed that these compounds all exhibited good HDAC1 inhibitory activity (Table 2), among which compound XC-6 (IC) showed the best performance. 50 = 0.0233μM), XC-10 (IC 50 = 0.0014μM), XC-11(IC 50 = 0.0507μM), XC-12 (IC 50 =0.0418μM), XC-14 (IC 50 = 0.0145μM), XC-21(IC 50 = 0.0078μM), XC-22(IC 50 = 0.0022μM), and XC-23 (IC 50 = 0.0443 μM) and other parameters showed superior performance compared to the positive control drug SAHA (vorinostat) (IC50).50 HDAC1 inhibitory activity (0.0723 μM) was measured.

[0226] Table 2. HDAC1 enzyme inhibitory activity of the target compound.

[0227]

[0228] 2. Western blotting assay to detect the effect of compounds on the STAT3 signaling pathway:

[0229] 2.1 Cell lines

[0230] MDA-MB-231 (human breast cancer cells).

[0231] 2.2 Experimental Procedure

[0232] First, wash and mix the cells with 1*PBS (Gibco, 10010023), aspirate, and add 80 μL of high-potency RIPA lysis buffer (Beyotime, P0013) to lyse the cells for 30 minutes. Centrifuge at 12000 rpm and 4°C for 15 minutes, collect 56 μL of supernatant, add 13 μL of loading buffer (Beyotime, P0015), heat at 95°C for 5 minutes, cool on ice, and load 15 μg onto a 10% SDS-PAGE gel. Run on a Bio-Rad protein electrophoresis apparatus at a constant voltage of 85V until the markers are separated, then switch to 120V and run to the bottom edge. After electrophoresis, remove the gel and cut a PVDF membrane to the appropriate size, in the following order from bottom to top: filter paper, PVDF membrane, gel, filter paper. Semi-dry transfer was performed at 12 V for 40 min to transfer the protein to a PVDF membrane (PerkinElmer, Northwalk, CT, USA). After transfer, the target band was cut and washed with 1*TBST (Beyotime, ST671) to remove Ponceau S. Blocking was performed at 37°C for 1 h with blocking buffer (1*TBST containing 5% w / v skim milk powder). After blocking, the membrane was washed three times with 15 ml of 1*TBST for 5 min each time. The primary antibody was diluted to the appropriate concentration recommended in the product instructions and placed in 10 ml of primary antibody dilution buffer (1*TBST containing 5% skim milk powder; to prepare 20 ml, add 1.0 g of skim milk powder to 20 ml of 1*TBST and mix thoroughly). The membrane was incubated overnight at 4°C with gentle agitation. After incubation, the membrane was washed three times with 10 ml of 1*TBST for 10 min each time. Then, the membrane was incubated with the developing solution (SignalFire™ ECLReagent #6883) at room temperature for 1 minute, the excess solution was discarded (the membrane was kept moist), and the membrane was exposed.

[0233] Test results are available Figure 1 , Figure 1The results showed that compounds XC-10 and XC-22 both had significant inhibitory effects on STAT3, inhibiting STAT3 phosphorylation in a concentration-dependent manner. Moreover, the inhibitory effect of compound XC-10 was comparable to that of the positive control drug Napabucasin.

[0234] 3. In vitro antitumor activity test of the target compound

[0235] 3.1 Sample preparation

[0236] After dissolving in DMSO (Merck), add PBS (-) to prepare a 1000 μM solution or a homogeneous suspension, and then dilute with PBS (-) containing DMSO.

[0237] 3.2 Cell lines

[0238] HCT116 (human colon cancer cells), MDA-MB-231 (human breast cancer cells), HepG-2 (human liver cancer cells), BxPC-3 (human pancreatic cancer cells), A549 (human lung cancer cells), and K562 (human chronic myeloid leukemia cells) were all cryopreserved and passaged in our laboratory.

[0239] 3.3 Culture medium

[0240] DMEM or PRMI1640 + 10% FBS + dual antibiotics

[0241] 3.4 Test Methods

[0242] MTT method: The concentration added per well of a 96-well plate is 3-6 × 10⁻⁶. 4 100 μL of cell suspension per cell / mL was incubated at 37 °C in a 5% CO2 incubator. After 24 hours, 100 µL of sample solution was added to each well, with triplet incubation, and the cells were incubated at 37 °C in a 5% CO2 incubator for 48 hours. 20 μL of 5 mg / mL MTT solution was added to each well, and the cells were incubated at 37 °C for another 2 hours. Then, 100 μL of DMSO was added to each well, and the cells were gently shaken for 10 min to mix thoroughly. The optical density (OD) of each well was measured at 570 nm using an ELISA reader. The experiment was repeated three times. The survival rate (%) was calculated using the following formula: Survival rate % = (OD value of experimental group / OD value of DMSO control group) × 100%.

[0243] Table 3. In vitro antiproliferative activity of the target compounds

[0244]

[0245] Table 4. In vitro antiproliferative activity of the target compounds

[0246]

[0247] As shown in Table 3, the Napabucasin derivatives involved in this invention exhibit in vitro inhibitory effects on both breast cancer cells and colorectal cancer cells. Compounds XC-10 and XC-22 showed significantly better inhibitory activity than the positive control drug Napabucasin. Furthermore, as shown in Table 4, compounds XC-10 and XC-22 also demonstrated good in vitro inhibitory effects on prostate cancer, leukemia, liver cancer, and lung cancer cells. The experimental results indicate that these dual-target compounds possess broad-spectrum antitumor activity.

[0248] The above description is merely a preferred embodiment of the present invention and is not intended to limit the present invention in any way. Although the present invention has been disclosed above with reference to preferred embodiments, it is not intended to limit the present invention. Any person skilled in the art can make some modifications or alterations to the above-described technical content to create equivalent embodiments without departing from the scope of the present invention. Any simple modifications, equivalent changes, and alterations made to the above embodiments based on the technical essence of the present invention without departing from the scope of the present invention shall still fall within the scope of the present invention.

Claims

1. A class of Napabucasin derivatives or pharmaceutically acceptable salts thereof possessing dual-target antitumor activity, characterized in that, The general structural formula of the Napabucasin derivative is shown below: in: A is selected from the following groups: hydroxyl, ... ; Where R1 is hydrogen, halogen, heterocyclic, R2 is hydrogen or halogen; The heterocycle is pyrroleyl, pyrazolyl, imidazolyl, furanyl, thiophenyl, oxazolyl, isoxazolyl, thiazolyl, isothiazolyl, pyridyl, pyrimidinyl, pyrazinyl, or pyridazinyl. R4 is selected from hydrogen, methyl, alkynyl, halogen, methoxy, trifluoromethyl, and cyano; Where X is selected from n is an integer selected from 1 to 10; R3 is selected from hydroxyl group, n is an integer selected from 1 to 6.

2. The Napabucasin derivative or its pharmaceutically acceptable salt with dual-target antitumor activity as described in claim 1, characterized in that, Pharmaceutically acceptable salts are their organic acid salts or inorganic acid salts.

3. The Napabucasin derivative or its pharmaceutically acceptable salt with dual-target antitumor activity according to claim 2, characterized in that, Inorganic acids include hydrochloric acid, sulfuric acid, phosphoric acid, diphosphoric acid, hydrobromic acid, or nitric acid; organic acids include acetic acid, maleic acid, fumaric acid, tartaric acid, succinic acid, lactic acid, p-toluenesulfonic acid, salicylic acid, oxalic acid, tannic acid, citric acid, trifluoroacetic acid, malic acid, or benzenesulfonate.

4. The Napabucasin derivative or its pharmaceutically acceptable salt with dual-target antitumor activity according to claim 1, characterized in that, The Napabucasin derivative is any one of the following: Compound XC-1: N-(2-aminophenyl)-4,9-dioxo-4,9-dihydronaphthoxy[2,3-b]furan-2-carboxamide, Compound XC-2: N-(3-((2-aminophenyl)amino)-3-oxopropyl)-4,9-dioxo-4,9-dihydronaphtho[2,3-b]furan-2-carboxamide, Compound XC-3: N-(2-amino-5-fluorophenyl)-4,9-dioxo-4,9-dihydronaphtho[2,3-b]furan-2-carboxamide, Compound XC-4: N-(2-amino-4,5-difluorophenyl)-4,9-dioxo-4,9-dihydronaphtho[2,3-b]furan-2-carboxamide, Compound XC-5: N-(4-amino-[1,1'-biphenyl]-3-yl)-4,9-dioxo-4,9-dihydronaphtho[2,3-b]furan-2-carboxamide, Compound XC-6: N-(4-amino-4'-fluoro-[1,1'-biphenyl]-3-yl)-4,9-dioxo-4,9-dihydronaphtho[2,3-b]furan-2-carboxamide, Compound XC-7: N-(4-amino-4'-methyl-[1,1'-biphenyl]-3-yl)-4,9-dioxo-4,9-dihydronaphtho[2,3-b]furan-2-carboxamide, Compound XC-8: N-(4-amino-4'-chloro-biphenyl-3-yl)-4,9-dioxo-4,9-dihydronaphtho[2,3-b]furan-2-carboxamide, Compound XC-9: N-(4-amino-4'-methoxy-biphenyl-3-yl)-4,9-dioxo-4,9-dihydronaphtho[2,3-b]furan-2-carboxamide, Compound XC-10: N-(2-amino-5-(pyridin-4-yl)phenyl)-4,9-dioxo-4,9-dihydronaphtho[2,3-b]furan-2-carboxamide, Compound XC-11: N-(2-amino-5-(pyridin-3-yl)phenyl)-4,9-dioxo-4,9-dihydronaphtho[2,3-b]furan-2-carboxamide, Compound XC-12: N-(2-amino-5-(furan-2-yl)phenyl)-4,9-dioxo-4,9-dihydronaphtho[2,3-b]furan-2-carboxamide, Compound XC-13: N-(2-amino-5-(thiophen-2-yl)phenyl)-4,9-dioxo-4,9-dihydronaphtho[2,3-b]furan-2-carboxamide, Compound XC-14: N-(2-amino-5-(thiophen-3-yl)phenyl)-4,9-dioxo-4,9-dihydronaphtho[2,3-b]furan-2-carboxamide, Compound XC-15: N-(4-amino-4'-(trifluoromethyl)-biphenyl-3-yl)-4,9-dioxo-4,9-dihydronaphtho[2,3-b]furan-2-carboxamide, Compound XC-16: N-(4-amino-4'-cyano-biphenyl-3-yl)-4,9-dioxo-4,9-dihydronaphtho[2,3-b]furan-2-carboxamide, Compound XC-17: N-hydroxy-4,9-dioxo-4,9-dihydronaphtho[2,3-b]furan-2-carboxamide, Compound XC-18: N-(4-(hydroxycarbamoyl)phenyl)-4,9-dioxo-4,9-dihydronaphtho[2,3-b]furan-2-carboxamide, Compound XC-19: N-(3-(hydroxyamino)-3-oxopropyl)-4,9-dioxo-4,9-dihydronaphtho[2,3-b]furan-2-carboxamide, Compound XC-20: N-(5-(hydroxyamino)-5-oxopentyl)-4,9-dioxo-4,9-dihydronaphtho[2,3-b]furan-2-carboxamide, Compound XC-21: N-(6-(hydroxyamino)-6-oxohexyl)-4,9-dioxo-4,9-dihydronaphtho[2,3-b]furan-2-carboxamide, Compound XC-22: N-(7-(hydroxyamino)-7-oxohepyl)-4,9-dioxo-4,9-dihydronaphtho[2,3-b]furan-2-carboxamide, Compound XC-23: N-(8-(hydroxyamino)-8-oxooctyl)-4,9-dioxo-4,9-dihydronaphtho[2,3-b]furan-2-carboxamide, Compound XC-24: N-(9-(hydroxyamino)-9-oxonyl)-4,9-dioxo-4,9-dihydronaphtho[2,3-b]furan-2-carboxamide, Compound XC-25: N'-ethyl-4,9-dioxo-4,9-dihydronaphtho[2,3-b]furan-2-carboxylhydrazine, Compound XC-26: 4,9-dioxo-N'-propyl-4,9-dihydronaphtho[2,3-b]furan-2-formylhydrazine, Compound XC-27: N'-butyl-4,9-dioxo-4,9-dihydronaphtho[2,3-b]furan-2-formylhydrazine, Compound XC-28: 4,9-dioxo-N'-pentyl-4,9-dihydronaphtho[2,3-b]furan-2-formylhydrazine, Compound XC-29: N'-isopentyl-4,9-dioxo-4,9-dihydronaphtho[2,3-b]furan-2-carboxylhydrazine.

5. A method for preparing a Napabucasin derivative or a pharmaceutically acceptable salt thereof with dual-target antitumor activity as described in claim 4, characterized in that, The synthetic methods for compounds XC-1 to XC-16 include the following steps: Compound 5 is condensed with compound 14 to give compound 15. Compound 15 is de-tert-butylated under acidic conditions to give compound 16. Compound 16 is condensed with o-phenylenediamine 17 protected by a Boc group to give compound 18. Compound 18 is de-Bocated under acidic conditions to give the target compound XC-2. Alternatively, compound 5 is condensed with o-phenylenediamine 19a-o with different substituents to give compound 20a-o. Compound 20a-o is de-Bocated under acidic conditions to give the target compound XC-1 or XC-3~16. The reaction route is as follows: Reagents and conditions: (a) HBTU, DIPEA, DMF, room temperature, 4 h, yield 64-74%; (b) TFA, room temperature, 1 h, yield 93-95%.

6. A method for preparing a Napabucasin derivative or a pharmaceutically acceptable salt thereof with dual-target antitumor activity as described in claim 4, characterized in that, The synthetic methods for compounds XC-17 to XC-24 include the following steps: Compound 5 is directly condensed with o-triphenylhydroxylamine 24 to give compound 26. Compound 26 is reacted with TFA to remove the protecting group to give the target compound XC-17; or, compound 5 is condensed with compound 21 to give compound 22. Compound 22 is de-tert-butylated under acidic conditions to give compound 23. Compound 23 is then condensed with o-triphenylhydroxylamine 24 to give compound 25. Compound 25 is reacted with TFA to remove the protecting group to give the target compound XC-18; or, compound 5 is condensed with alkane chains of different lengths to give compounds 28a-f. Compound 28a-f is de-tert-butylated under acidic conditions to give compounds 29a-f. Compound 29a-f is then condensed with o-triphenylhydroxylamine 24 to give compounds 30a-f. Compound 25 is reacted with TFA to remove the protecting group to give the target compounds XC-19~24. The reaction route is as follows: Reagents and conditions: (a) HBTU, DIPEA, DMF, room temperature, 4 h, yield 64-74%; (b) TFA, room temperature, 1 h, yield 93-95%.

7. A method for preparing a Napabucasin derivative or a pharmaceutically acceptable salt thereof with dual-target antitumor activity as described in claim 4, characterized in that, The synthetic methods for compounds XC-25 to XC-29 include the following steps: Compound 5 is condensed with Boc-protected hydrazides of different chain lengths 31a-d to generate compounds 32a-d. Compound 32a-d is de-tert-butylated under acidic conditions to give the target compounds XC-25~28. Alternatively, compound 5 is condensed with compound 33 to generate compound 34. Compound 34 is de-tert-butylated under acidic conditions to give the target compound XC-29. The reaction route is as follows: Reagents and conditions: (a) HBTU, DIPEA, DMF, room temperature, 4 h, yield 64-74%; (b) TFA, room temperature, 1 h, yield 93-95%.

8. The use of a Napabucasin derivative or a pharmaceutically acceptable salt thereof with dual-target antitumor activity as described in claim 1 in the preparation of antitumor drugs or drugs for differentiation and proliferation-related diseases.

9. The application according to claim 8, characterized in that, The tumor is selected from colorectal cancer, breast cancer, lung cancer, liver cancer, and prostate cancer. The medicinal salt is anhydrous or contains one or more molecules of water of crystallization.

10. The use of a Napabucasin derivative or a pharmaceutically acceptable salt thereof with dual-target antitumor activity as described in claim 1 in the preparation of HDAC inhibitors, STAT3 inhibitors or HDAC / STAT3 dual-target inhibitors.

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