Aryl ketone derivative and furazan spliced conjugate as well as preparation method and application thereof

Abstract

The invention relates to the field of chemical pharmacy, in particular to an aryl ketone derivative and furazan spliced conjugate as well as a preparation method and application thereof. The structural formula of the conjugate spliced by the aryl ketone derivative and the furazan is as shown in any one of formulas I-V: # imgabs0 # imgabs1 #. A coumarin skeleton is opened, and the conjugate spliced by the aryl ketone derivative and the furazan is designed and synthesized. The aryl ketone derivative and furazan combined conjugate disclosed by the invention is novel in structure, has the effect of highly selectively inhibiting the proliferation activity of P-gp overexpression drug-resistant tumor cells, and provides a new thought and direction for overcoming the multidrug resistance of the tumor cells caused by P-gp overexpression.

Description

Technical Field

[0001] The present invention relates to the field of chemical pharmaceuticals, and particularly to a conjugate formed by the combination of an aryl ketone derivative and furazan, and a preparation method and application thereof. Background Art

[0002] Nitric Oxide (NO) is an important signaling molecule that can participate in the regulation of various physiological functions and also affect the growth, proliferation, and metastasis of tumor cells. Research has shown that low concentrations of NO can promote tumor angiogenesis and thus promote tumor cell proliferation, while high concentrations of NO can inhibit tumor cell growth by causing DNA damage and inducing apoptosis. Therefore, the research on anti-tumor drugs based on NO donors has received extensive attention (Journal of Medicinal Chemistry, 2017, 60: 7617). Summary of the Invention

[0003] The object of the present invention is to provide a novel anti-tumor compound (a conjugate formed by the combination of an aryl ketone derivative and furazan) with clear activity, good water solubility, and high selective inhibitory effect on MDR tumor drug-resistant strains overexpressing P-gp.

[0004] To achieve the above object, the present invention provides the following solutions:

[0005] The present invention provides a conjugate formed by the combination of an aryl ketone derivative and furazan, and the structural formula is shown as any one of Formulas I to V:

[0006]

[0007] In Formulas I to V, R2, R3, R4, and R5 are independently one of H, F, OCH3, CN, NH2, CF3, and OH; n = 2 or 3.

[0008] In a preferred embodiment of the present invention, in Formula I, R2 = H, F, OCH3, or CN;

[0009] In Formula II, R3 = H, F, or OCH3;

[0010] In Formula IV, R4 = H, F, NH2, CF3, CN, or OH;

[0011] In Formula V, R5 = H, F, NH2, CF3, or CN.

[0012] The present invention also provides a preparation method of the above conjugate formed by the combination of an aryl ketone derivative and furazan. When the structural formula of the conjugate formed by the combination of an aryl ketone derivative and furazan is shown as Formula I, the preparation method of the conjugate formed by the combination of an aryl ketone derivative and furazan includes the following steps:

[0013] Using 2,4-dihydroxyacetophenone as the raw material, the 4-hydroxy group in the raw material was protected with dihydropyran under the catalysis of 4-toluenesulfonic acid pyridine salt to obtain compound 2; compound 2 was methylated with CH3I to generate compound 3, and then underwent aldol condensation reaction with benzaldehydes containing different substituents, and the pyran protecting group was removed under hydrochloric acid to synthesize intermediates 4a-d;

[0014] In the presence of K2CO3 and NaI, intermediates 4a-d were etherified with 2-bromoethanol or 3-bromo-1-propanol to obtain compounds 5a-f;

[0015] At room temperature, in dichloromethane solution and under the catalytic condition of 1,8-diazabicyclo[5.4.0]-7-undecene, compounds 5a-f were coupled with phenylsulfonyl furazan N-oxide to obtain the conjugate of the aryl ketone derivative shown in formula I and furazan;

[0016] When the conjugate of the aryl ketone derivative and furazan has the structural formula shown in formula II, the preparation method of the conjugate of the aryl ketone derivative and furazan includes the following steps:

[0017] The double bond in the α,β-unsaturated ketone side chain of intermediates 4a-d was locally reduced with palladium-carbon in phenyl sulfide / methanol to synthesize intermediates 7a-c, and then etherified with 2-bromoethanol or 3-bromo-1-propanol to obtain compounds 8a-f; at room temperature, under the catalysis of DBU / DCM, compounds 8a-f were coupled with phenylsulfonyl furazan N-oxide to obtain the conjugate of the aryl ketone derivative shown in formula II and furazan;

[0018] When the conjugate of the aryl ketone derivative and furazan has the structural formula shown in formula III, the preparation method of the conjugate of the aryl ketone derivative and furazan includes the following steps:

[0019] The pyran protecting group of compound 3 was removed to obtain 4-hydroxy-2-methoxyacetophenone; 4-hydroxy-2-methoxyacetophenone was refluxed with bromoethanol or bromopropanol, potassium carbonate and sodium iodide in DMF to obtain 4-hydroxyethoxy- or hydroxypropoxy-substituted 2-methoxyacetophenone, and finally reacted with phenylsulfonyl furazan N-oxide to obtain the conjugate of the aryl ketone derivative shown in formula III and furazan;

[0020] When the conjugate of the aryl ketone derivative and furazan has the structural formula shown in formula IV, the preparation method of the conjugate of the aryl ketone derivative and furazan includes the following steps:

[0021] In ethanol, compound 13 reacts with bromoethanol, sodium hydroxide and sodium iodide under stirring to obtain 4-hydroxyethoxysalicylic acid; 4-hydroxyethoxysalicylic acid condenses with substituted ampicillin to form amide side chain intermediates 15a-f, and then reacts with phenylsulfonylfurazan N-oxide in DCM containing DBU at room temperature to synthesize the conjugate of the aryl ketone derivative shown in formula IV and furazan;

[0022] The structural formula of the said compound 13 is

[0023] When the structural formula of the conjugate of the aryl ketone derivative and furazan is as shown in formula V, the preparation method of the conjugate of the aryl ketone derivative and furazan includes the following steps:

[0024] Intermediates 15a-b and 15d-e are first etherified with CH3I under the catalysis of potassium carbonate to form 2-methoxy derivatives, and then react with phenylsulfonylfurazan N-oxide in DCM containing DBU at room temperature to synthesize the conjugate of the aryl ketone derivative shown in formula V and furazan.

[0025] The present invention also provides an application of the conjugate of the above-mentioned aryl ketone derivative and furazan in the preparation of anti-tumor drugs.

[0026] In a preferred embodiment of the present invention, the tumor is caused by human triple-negative breast cancer cells MDA-MB-231 and MDA-MB-468, breast cancer cells MCF-7 or drug-resistant tumor cells MCF-7 / ADR.

[0027] The conjugate of the aryl ketone derivative and furazan of the present invention selectively inhibits the proliferation of MDR tumor cells with overexpressed P-gp through the released nitric oxide.

[0028] The present invention also provides an anti-tumor drug, and the active ingredient includes the conjugate of the above-mentioned aryl ketone derivative and furazan.

[0029] The present invention opens the coumarin skeleton and designs and synthesizes the conjugate of the aryl ketone derivative and furazan. Its water solubility is increased by 24-775 times compared with CY-16S-4A93. The evaluation of the proliferation inhibitory activity against four breast cancer cells MCF-7, MCF-7 / ADR, MDA-MB-231, and MDA-MB-468 shows that most compounds exhibit excellent anti-proliferation activity against the adriamycin-resistant breast cancer cell line MCF-7 / ADR, and the proliferation inhibitory activity against MCF-7 / ADR is 7-1716 times higher than that against the sensitive strain MCF-7. Most compounds also have good inhibitory activity against MDA-MB-468, IC 50It is between 80 - 780 nM. In addition, the active compounds show low toxicity to normal mammary epithelial cells MCF-10A and human umbilical vein endothelial cells HUVEC. Preliminary pharmacological mechanism studies have shown that such target compounds can enter the lysosomes of drug-resistant MCF-7 / ADR cells as potential P-glycoprotein (P-gp) substrates, release nitric oxide, generate reactive oxygen species, and promote tumor cell apoptosis. At the same time, they can also inhibit tumor cell proliferation through multiple pathways such as inducing autophagy and ferroptosis in MCF-7 / ADR cells. This lays a foundation for the subsequent research and development of anti-tumor candidate compounds with novel structures and special pharmacological action mechanisms to overcome multi-drug resistance in breast cancer.

[0030] The present invention discloses the following technical effects:

[0031] The conjugate of the aryl ketone derivative and furazan in the present invention has a novel structure and has the effect of highly selectively inhibiting the proliferation activity of P-gp overexpressing drug-resistant tumor cells, which will provide new ideas and directions for overcoming the multi-drug resistance of tumor cells caused by P-gp overexpression. Brief Description of the Drawings

[0032] In order to more clearly illustrate the technical solutions in the embodiments of the present invention or the prior art, the following will briefly introduce the drawings required for use in the embodiments. Obviously, the drawings in the following description are only some embodiments of the present invention. For those of ordinary skill in the art, without creative efforts, other drawings can also be obtained based on these drawings.

[0033] Figure 1 It is the design of the conjugate of the aryl ketone derivative and furazan in the present invention.

[0034] Figure 2 It is the inhibition of HUVEC cell proliferation by 6 preferred target compounds of the present invention at a concentration of 500 nM.

[0035] Figure 3 It is the release of NO by the active compound 9e of the present invention in the lysosomes of MCF-7 / ADR and MCF-7 cells.

[0036] Figure 4 It is the induction of autophagy in drug-resistant breast cancer MCF-7 / ADR cells by the active compound 9e of the present invention.

[0037] Figure 5 It is the promotion of ferroptosis in MCF-7 / ADR cells by the active compound 9e of the present invention. Detailed Embodiments

[0038] The various exemplary embodiments of the present invention will be described in detail below. This detailed description should not be considered as a limitation on the present invention, but rather as a more detailed description of certain aspects, features, and implementation schemes of the present invention.

[0039] It should be understood that the terms used in the present invention are only for describing specific embodiments and are not intended to limit the present invention. Additionally, for the numerical ranges in the present invention, it should be understood that each intermediate value between the upper and lower limits of the range is also specifically disclosed. Any intermediate value within any stated value or stated range, as well as each smaller range between any other stated value or intermediate value within the stated range, is also included in the present invention. The upper and lower limits of these smaller ranges may be independently included or excluded from the range.

[0040] Unless otherwise specified, all technical and scientific terms used herein have the same meaning as commonly understood by those of ordinary skill in the art to which the present invention pertains. Although the present invention only describes preferred methods and materials, any methods and materials similar or equivalent to those described herein may also be used in the implementation or testing of the present invention. All documents mentioned in this specification are incorporated by reference to disclose and describe the methods and / or materials related to the said documents. In case of conflict with any incorporated document, the content of this specification shall prevail.

[0041] Without departing from the scope or spirit of the present invention, various improvements and changes can be made to the specific embodiments of the present invention specification, which are obvious to those skilled in the art. Other embodiments obtained from the specification of the present invention are obvious to those skilled in the art. The specification and examples of the present invention are merely exemplary.

[0042] Regarding the use of "comprising", "including", "having", "containing", etc. in this article, they are all open-ended terms, meaning including but not limited to.

[0043] Based on the original active compound CY-16S-4A93, the present invention uses the skeleton cleavage strategy to open the coumarin lactone ring and replaces it with α,β-unsaturated ketone, α,β-saturated ketone, and amide side chains, and designs and synthesizes Figure 1 the conjugate of aryl ketone derivative and furazan as shown, which has significant inhibitory activity against the proliferation of tumor cells such as MCF-7 / ADR.

[0044] In the present invention, the preferred aryl ketone-furazan derivatives have the structures of Formulas I, II, III, IV, and V. The benzene ring at the 3-position of the coumarin lactone ring-opened structure is substituted with a methoxy group, and the 4-position is linked with a phenyl group containing a para-substituent through an α,β-unsaturated ketone side chain. The 1-position is combined with furazan N-oxide through a 2- or 3-carbon side chain to form the product in Type I; the benzene ring at the 3-position of the coumarin parent nucleus lactone ring-opened structure is substituted with a methoxy group, and the 4-position is linked with a phenyl group containing a para-substituent through an α,β-saturated ketone side chain. The 1-position is combined with furazan N-oxide through a 2- or 3-carbon side chain to form the product in Type II; after the coumarin ring is opened, the benzene ring at the 3-position is substituted with a methoxy group, and the 4-position is substituted with an acetyl group. The 1-position is combined with furazan N-oxide through a 2- or 3-carbon side chain to form the product in Type III; the benzene ring at the 3-position of the coumarin parent nucleus lactone ring-opened structure is substituted with a hydroxyl group, and the 4-position is linked with a phenyl group containing a para-substituent through an amide side chain. The 1-position is combined with furazan N-oxide through a 2-carbon side chain to form the product in Type IV; and the benzene ring at the 3-position is substituted with a methoxy group, and the 4-position is linked with a phenyl group containing a para-substituent through an amide side chain. The 1-position is combined with furazan N-oxide through a 2-carbon side chain to form the product in Type V. (As Figure 1 shown), where: R1 is furazan N-oxide; R is a hydrogen atom, a methoxy group, a trifluoromethoxy group, a fluorine atom, a cyano group, a hydroxyl group, an amino group, a tert-butoxycarbonylamino group; n is 2 or 3.

[0045] The present invention provides a method for preparing the conjugate of the aryl ketone derivative and furazan, which includes:

[0046] Using 2,4-dihydroxyacetophenone (1) as a raw material, the 4-position hydroxyl group in the raw material is protected with dihydropyran (DHP) under the catalysis of 4-toluenesulfonic acid pyridinium salt (PPTS) to obtain Compound 2. Then it is methylated with CH3I to generate 3, and then undergoes an aldol condensation reaction with several benzaldehydes containing different substituents. The pyran protecting group is removed in 4M hydrochloric acid to synthesize the phenyl ketone intermediate 4a-d with an α,β-unsaturated ketone side chain. In the presence of K2CO3 and NaI, the intermediate 4a-d is etherified with 2-bromoethanol or 3-bromo-1-propanol to obtain 5a-f. At room temperature, in dichloromethane (DCM) solution, under the catalysis of 1,8-diazabicyclo[5.4.0]-7-undecene (DBU), it is coupled with benzenesulfonyl furazan N-oxide to obtain the aryl ketone / furazan target compound 6a-f with an α,β-unsaturated ketone side chain. The chemical reaction is as shown in General Formula I:

[0047]

[0048] The double bond in the α,β-unsaturated ketone side chain of the above compounds 4a-c was locally reduced with palladium-carbon in phenyl sulfide / methanol to synthesize intermediates 7a-c, which were then etherified with 2-bromoethanol or 3-bromo-1-propanol to obtain 8a-f. At room temperature, they were coupled with phenylsulfonyl furazan N-oxide under the catalysis of DBU / DCM to obtain aryl ketone / furazan target compounds 9a-f containing α,β-saturated ketone side chains. The chemical reaction is shown in General Formula II:

[0049]

[0050] Compound 3 was deprotected from the pyran protecting group with 4 equivalents of hydrochloric acid in methanol solvent to obtain 4-hydroxy-2-methoxyacetophenone (10), which was then refluxed with bromoethanol or bromopropanol, potassium carbonate, and sodium iodide in DMF to obtain 4-hydroxyethoxy- or hydroxypropoxy-substituted 2-methoxyacetophenone (11a-b). Finally, it was reacted with phenylsulfonyl furazan N-oxide at room temperature in dichloromethane (DCM) containing DBU to synthesize acetophenone / furazan target derivatives 12a-b. The chemical reaction is shown in General Formula III:

[0051]

[0052] In ethanol, compound 13 was stirred and reacted with bromoethanol, sodium hydroxide, and sodium iodide at 60 °C to obtain 4-hydroxyethoxysalicylic acid 14, which was then condensed with substituted ampicillin under the catalysis of DCC to form amide side chain intermediates 15a-f. Then, it was reacted with phenylsulfonyl furazan N-oxide in DCM containing DBU at room temperature to synthesize aryl ketone / furazan target derivatives 16a-f containing 2-hydroxy-1-amide side chain substitution. The chemical reaction is shown in General Formula IV:

[0053]

[0054] The hydroxyl groups in the structures of intermediates 15a-b and 15d-e in General Formula IV were first etherified with CH3I under the catalysis of potassium carbonate to form 2-methoxy derivatives 17a-b and 17d-e, and then reacted with phenylsulfonyl furazan N-oxide in DCM containing DBU at room temperature to synthesize aryl ketone / furazan target derivatives 18a-b and 18d-e containing 2-methoxy-1-amide side chain substitution. The chemical reaction is shown in General Formula V:

[0055]

[0056] In the present invention, a series of conjugates (6a-f, 9a-f, 12a-b, 16a-f, 18a-b, 18d-e) are synthesized by opening the lactone ring of coumarin and replacing it with α,β-unsaturated and saturated ester side chains and amide side chains, and then coupling them with furazan N-oxide through a 2-3 carbon linker. Solubility determination, anti-tumor activity screening at the cellular level, and preliminary pharmacological mechanism studies are carried out on these conjugates.

[0057] In vitro pharmacological experiments were conducted with CY-16S-4A93 and doxorubicin as positive drug controls to test the proliferation inhibitory activities of the target compounds against breast cancer cell lines MCF-7, doxorubicin-resistant breast cancer cell line MCF-7 / ADR, and two triple-negative breast cancer cell lines MDA-MB-231 and MDA-MB-468. The results showed that most of the target compounds exhibited excellent anti-proliferative activity against MCF-7 / ADR. Among them, the open-ring target compounds with α,β-saturated side chains had the best selective inhibition. The proliferation inhibitory activities of compounds 9a and 9e against MCF-7 / ADR were 1716 and 1401 times higher than those against MCF-7, far higher than the 499-fold of the coumarin backbone control CY-16S-4A93. In addition, the target compounds 6c-d and 9e-f showed low cell proliferation inhibitory activities against normal breast epithelial cells MCF-10A and human umbilical vein endothelial cells HUVEC, indicating good safety.

[0058] Preliminary pharmacological mechanism studies showed that the target compound 9e is a potential P-gp substrate, which can release an effective concentration of nitric oxide in the lysosomes of MCF-7 / ADR, generate reactive oxygen species, and induce apoptosis of tumor cells. At the same time, laser confocal microscopy and Western Blot experiments showed that compound 9e could induce autophagy in MCF-7 / ADR cells. In addition, the ferroptosis inhibitor Fer-1 could completely abolish the anti-tumor activity of compound 9e, indicating that 9e may also inhibit tumor cell proliferation by promoting ferroptosis. This laid a foundation for the subsequent development of novel nitric oxide donor anti-tumor candidate compounds with special pharmacological mechanisms to overcome breast cancer multidrug resistance.

[0059] The method for studying the biological activities of the conjugates of aryl ketone derivatives and furazan in the present invention is as follows:

[0060] 1. Use the MTS method to determine the cytotoxicity of sensitive and drug-resistant solid tumor cells. Set up blank control groups, target compound groups (6a-f, 9a-f, 12a-b, 16a-f, 18a-b, 18d-e), compound CY-16S-4A93 group, and positive drug (doxorubicin) control groups. After adding the drugs, incubate at 37 °C for 48 hours, perform MTS detection, and calculate the inhibition rate and IC 50Values. Tested tumor cell lines: MCF-7 (human breast cancer cell line) and MCF-7 / ADR (adriamycin-resistant breast cancer cell line), MDA-MB-231 and MDA-MB-468 (two human triple-negative breast cancer cell lines).

[0061] 2. Use the MTS method to test the IC 50 values of the target compound against non-tumor cells of HUVEC (human umbilical vein endothelial cell line) and MCF-10A (human normal mammary epithelial cell line), verify whether the target compound is toxic to normal cells, and evaluate its safety.

[0062] 3. Use DAF-FM DA probe, Hoechst 33342 and Lyso-NO probe to observe the nitric oxide release of the target compound in MCF-7 / ADR tumor cells and lysosomal subcellular compartments.

[0063] 4. Simultaneously perform immunofluorescence staining of cells with LC3 and LAMP1 antibodies, and observe autophagy by laser confocal microscopy; and use the MTT method to observe the ferroptosis phenomenon induced by the target compound by co-incubating with the ferroptosis inhibitor Fer-1.

[0064] The technical solutions described in the present invention are all conventional solutions in the art if not otherwise specified. The reagents or raw materials used are all purchased from commercial channels or are publicly available if not otherwise specified.

[0065] The technical solutions provided by the present invention will be described in detail below in conjunction with the embodiments, but they should not be construed as limiting the protection scope of the present invention.

[0066] Example 1

[0067] Synthesis of compound 1-(2-hydroxy-4-((tetrahydro-2H-pyran-2-yl)oxy)phenyl)ethan-1-one (2) in general formula I

[0068] 2,4-Dihydroxyacetophenone (7.6 g, 50 mmol, 1 eq.) and pyridinium 4-methylbenzenesulfonate (PPTS, 0.5 g, 2 mmol, 0.04 eq.) were placed in a 250 mL flask, dissolved in dichloromethane (70 mL), and then 3,4-dihydro-2H-pyran (DHP, 11.6 mL, 2.5 eq.) was added. The reaction was stirred at room temperature for 12 h. After the reaction was monitored by TLC (PE:EA = 10:1) to completion, it was washed with saturated NaHCO3 solution. The organic phase was washed three times with saturated brine (50 mL), dried over anhydrous sodium sulfate, and concentrated under reduced pressure by rotary evaporation to a small amount of liquid. Petroleum ether (PE, 40 mL) was added to the concentrated solution, and after ultrasonic treatment, it was filtered. The solid surface was rinsed with a small amount of a mixed solution of ethyl acetate and dichloromethane, and dried in an infrared oven. Finally, white solid 2 (9.75 g) was obtained with a yield of 82.5%. 1H NMR (400 MHz, CDCl3) δ 12.61 (s, 1H, -ArOH), 7.63 (d, J = 8.7 Hz, 1H, 6-ArH), 6.65 - 6.50 (m, 2H, 3,5-ArH), 5.48 (t, J = 3.0 Hz, 1H, -OCH-O-), 3.88 - 3.56 (d, 2H, -CH2CH2O-), 2.56 (s, 3H, -COCH3), 2.07 - 1.47 (m, 6H, -CH2-). ESI-MS m / z 237.4 [M + H] + .。

[0069] Example 2

[0070] Synthesis of compound 1-(2-methoxy-4-((tetrahydro-2H-pyran-2-yl)oxy)phenyl)ethan-1-one (3) in general formula I

[0071] Compound 2 (2.37 g, 12 mmol, 1 eq.) was placed in a 50 mL flask, N,N-dimethylformamide (10 mL) and potassium carbonate (8.29 g, 60 mmol, 5 eq.) were added, and the temperature was raised to 80 °C with stirring; after 0.5 h, methyl iodide solution (1.5 mL, 2 eq.) was slowly added, and the reaction was stirred at 80 °C for 1.5 h. The heating was stopped and the reaction solution was cooled to room temperature. Ethyl acetate (EA, 20 mL) was added, and part of the K2CO3 was removed by filtration. Water (40 mL) was added to the filtrate, and it was extracted with EA. The organic phase was collected, washed three times with saturated brine (50 mL), dried over anhydrous sodium sulfate, and concentrated under reduced pressure by rotary evaporation to obtain crude product 3 (2.61 g), which was a yellow liquid and could be directly used for the next reaction without purification.

[0072] Example 3

[0073] Synthesis of compounds 4a-d in general formula I

[0074] The crude product 3 (5 mmol, 1 eq.) was added to methanol (10 mL), and benzaldehyde with the corresponding substituent (12 mmol, 2.4 eq.) and sodium hydroxide (480 mg, 12 mmol, 2.4 eq.) were added. The temperature was raised to 40 °C and stirred for 1.5 days. Subsequently, the pH was adjusted to below 3 with 4 M hydrochloric acid and stirred at room temperature for 12 h. The reaction solution was concentrated under reduced pressure, then extracted with water (20 mL) and ethyl acetate (20 mL × 3). The collected organic phase was washed three times with saturated brine, dried over anhydrous sodium sulfate, concentrated by distillation under reduced pressure to precipitate some solids, 25 mL of a mixed solution of petroleum ether and ethyl acetate (PE:EA = 4:1) was added, sonicated and then filtered by suction. The solid was rinsed with a small amount of the mixed solution of PE and EA (PE:EA = 4:1), placed in an infrared oven to dry, and intermediate 4a-d could be obtained. The mother liquor was purified by column chromatography (PE:EA = 3:1) to obtain the pure product for structure identification.

[0075] 4a ((E)-1-(4-Hydroxy-2-methoxyphenyl)-3-phenylprop-2-en-1-one): Yield 87.9%, light yellow solid; 1 H NMR (400 MHz, DMSO-d6) δ 13.51 (s, 1H, -ArOH), 7.81–7.66 (m, 3H), 7.60 (m, 2H), 7.54 (d, J = 15.6 Hz, 1H), 7.43-7.37 (m, 2H), 6.56-6.45 (m, 2H), 3.84 (s, 3H). ESI-MS m / z 255.1 [M+H] + .

[0076] 4b ((E)-3-(4-Fluorophenyl)-1-(4-hydroxy-2-methoxyphenyl)prop-2-en-1-one): Yield 78.8%, yellow solid; 1 H NMR (400 MHz, CDCl3) δ 7.70-7.62 (m, 2H), 7.59-7.55 (m, 2H), 7.43 (d, J = 15.8 Hz, 1H), 7.07 (t, J = 8.6 Hz, 2H), 6.53-6.44 (m, 2H), 3.84 (s, 3H). ESI-MS m / z 272.8 [M+H] + .

[0077] 4c ((E)-1-(4-Hydroxy-2-methoxyphenyl)-3-(4-methoxyphenyl)prop-2-en-1-one): Yield 92.5%, white solid; 11H NMR (400 MHz, CDCl3) δ 7.73 - 7.65 (m, 2H), 7.56 (d, J = 8.3 Hz, 2H), 7.39 (d, J = 15.6 Hz, 1H), 6.92 (d, J = 8.3 Hz, 2H), 6.54 - 6.47 (m, 2H), 3.88 (s, 3H), 3.85 (s, 3H). ESI-MS m / z 285.3 [M+H] + .

[0078] 4d ((E)-4-(3-(4-Hydroxy-2-methoxyphenyl)-3-oxopropyl-1-en-1-yl)benzonitrile): Yield 74.2%, pale yellow solid. 1 1H NMR (400 MHz, CDCl3) δ 8.06 (d, J = 8.3 Hz, 1H), 7.72 (d, J = 8.3 Hz, 1H), 7.69 - 7.63 (m, 5H), 6.55 - 6.48 (m, 2H), 3.89 (s, 3H). ESI-MS m / z 279.9 [M+H] + .

[0079] Example 4

[0080] Synthesis of Compounds 5a-f in General Formula I

[0081] Place compounds 4a-d (1.5 mmol, 1 eq.), potassium carbonate (622 mg, 4.5 mmol, 3 eq.) and sodium iodide (23 mg, 0.15 mmol, 0.1 eq.) in a 25 mL flask, add DMF (3 mL), heat to 90 °C, and inject bromoethanol (or bromopropanol) (1.95 mmol, 1.3 eq.) with a syringe. Keep the reaction at 90 °C for 18 h. Monitor the reaction by TLC until completion (PE:EA = 2:1). Add ethyl acetate (10 mL), stir for 15 min, add water (20 mL), and extract repeatedly with ethyl acetate. Wash the collected organic phase four times with saturated brine to remove DMF in the reaction system as much as possible. Dry over anhydrous sodium sulfate and concentrate under reduced pressure by rotary evaporation to obtain the crude product 5a-f. The product is directly used for the next reaction without purification.

[0082] Example 5

[0083] Synthesis of Target Compounds 6a-f in General Formula I

[0084] Compounds 5a-f (1.05 mmol, 1 eq.) and phenylsulfonyl furazan N-oxide (1.35 mmol, 1.3 eq.) were placed in a reaction flask, immersed in an ice-water bath, anhydrous dichloromethane (10 mL) was added, and after stirring for 10 min, 1,8-diazabicyclo[5.4.0]undec-7-ene (DBU, 2.1 mmol, 2 eq.) was slowly added. After stirring for another 15 min, the ice bath was removed and the reaction was carried out at room temperature for 16 - 20 h. The reaction was monitored by TLC (PE:EA = 2:1) and was basically complete. It was extracted three times with DCM (15 mL), the organic phase was washed twice with saturated brine, dried over anhydrous sodium sulfate, and concentrated by rotary evaporation under reduced pressure to obtain a crude solid. After purification by column chromatography (DCM:PE = 3:1), the target compounds 6a-f were obtained.

[0085] 6a (4-(3-(4-Cinnamoyl-3-methoxyphenoxy)propoxy)-3-(phenylsulfonyl)-1,2,5-oxadiazole 2-oxide): Yield 25%, white solid. 1 H NMR (400 MHz, CDCl3) δ8.02–7.95 (m, 2H, 2”,6”-ArH), 7.78 (d, J = 8.6 Hz, 1H, 6-ArH), 7.75–7.70 (m, 1H, 4”-ArH), 7.69 (d, J = 15.8 Hz, 1H, Ar CH =CH-), 7.62–7.58 (m, 2H, 3”,5”-ArH), 7.57–7.52 (m, 2H, 2’,6’-ArH), 7.53 (d, J = 16.0 Hz, 1H, ArCH= CH -), 7.45–7.36 (m, 3H, 3’,4’,5’-ArH), 6.60 (dd, J1 = 8.6 Hz, J2 = 2.2 Hz, 1H, 5-ArH), 6.55 (d, J = 2.2 Hz, 1H, 3-ArH), 4.66 (t, J = 6.0 Hz, 2H, -O CH2 CH2CH2O-), 4.27 (t, J = 5.8 Hz, 2H, -OCH2CH2 CH2 O-), 3.91 (s, 3H, -OCH3), 2.40 (p, J = 5.9 Hz, 2H, -OCH2 CH2 CH2O-). ESI-MS m / z 338.6 [M+H] + . ESI-MS m / z 537.1 [M+H] + .

[0086] 6b ((E)-4-(3-(3-(4-(4-Fluorophenyl)acryloyl)-3-methoxyphenoxy)propoxy)-3-(phenylsulfonyl)-1,2,5-oxadiazole 2-oxide): Yield 17%, white solid. 1 H NMR (400 MHz, CDCl3) δ 8.03–7.96 (m, 2H, 2”,6”-ArH), 7.79 (d, J = 8.5 Hz, 1H, 6-ArH), 7.73 (t, J = 7.0 Hz, 1H, 4”-ArH), 7.66 (d, J = 15.8 Hz, 1H, Ar CH =CH-), 7.62–7.51 (m, 4H, 2’,6’,3”,5”-ArH), 7.46 (d, J = 15.8 Hz, 1H, ArCH= CH -), 7.09 (t, J = 8.7 Hz, 2H, 3’,5’-ArH), 6.61 (dd, J1 = 8.6 Hz, J2 = 2.2 Hz, 1H, 5-ArH), 6.55 (d, J = 2.2 Hz, 1H, 3-ArH), 4.67 (t, J = 6.0 Hz, 2H, -O CH2 CH2CH2O-), 4.27 (t, J = 5.8 Hz, 2H, -OCH2CH2 CH2 O-), 3.92 (s, 3H, -OCH3), 2.41 (p, J = 5.9 Hz, 2H, -OCH2 CH2 CH2O-). ESI-MS m / z 554.9 [M+H] + .

[0087] 6c ((E)-4-(2-(3-Methoxy-4-(3-(4-methoxyphenyl)acryloyl)phenoxy)ethoxy)-3-(phenylsulfonyl)-1,2,5-oxadiazole 2-oxide): Yield 37%, white solid. 1 H NMR (400 MHz, CDCl3) δ 8.01 (d, J = 8.6 Hz, 2H, 2”,6”-ArH), 7.76 (d, J = 8.5 Hz, 1H, 6-ArH), 7.74–7.69 (m, 1H, 4”-ArH), 7.66 (d, J = 15.7 Hz, 1H, Ar CH =CH-), 7.59–7.50 (m, 4H, 2’,6’,3”,5”-ArH), 7.37 (d, J = 15.8 Hz, 1H, ArCH= CH-), 6.92 (d, J = 8.9 Hz, 2H, 3’,5’-ArH), 6.58 (dd, J1 = 8.5 Hz, J2 = 2.2 Hz, 1H, 5-ArH), 6.56 (d, J = 2.0 Hz, 1H, 3-ArH), 4.80 (t, J = 4.3 Hz, 2H, -O CH2 CH2O-), 4.46 (t, J = 4.5 Hz, 2H, -OCH2 CH2 O-), 3.92 (s, 3H, 2-OCH3), 3.85 (s, 3H, 4’-OCH3). ESI-MS m / z 553.2 [M+H] + .

[0088] 6d ((E)-4-(3-(3-Methoxy-4-(3-(4-methoxyphenyl)acryloyl)phenoxy)propoxy)-3-(phenylsulfonyl)-1,2,5-oxadiazole 2-oxide): Yield 33%, white solid. 1 H NMR (400 MHz, CDCl3) δ 7.98 (d, J = 8.6 Hz, 2H, 2”,6”-ArH), 7.75 (d, J = 8.6 Hz, 1H, 6-ArH), 7.74–7.69 (m, 1H, 4”-ArH), 7.66 (d, J = 15.7 Hz, 1H, Ar CH =CH-), 7.59–7.49 (m, 4H, 2’,6’,3”,5”-ArH), 7.39 (d, J = 15.8 Hz, 1H, ArCH= CH -), 6.92 (d, J = 8.6 Hz, 2H, 3’,5’-ArH), 6.59 (dd, J1 = 8.6 Hz, J2 = 2.2 Hz, 1H, 5-ArH), 6.54 (d, J = 2.1 Hz, 1H, 3-ArH), 4.66 (t, J = 6.0 Hz, 2H, -O CH2 CH2CH2O-), 4.26 (t, J = 5.8 Hz, 2H, -OCH2CH2 CH2 O-), 3.90 (s, 3H, 2-OCH3), 3.85 (s, 3H, 4’-OCH3), 2.40 (p, J = 5.9 Hz, 2H, -OCH2 CH2 CH2O-). ESI-MS m / z 567.4 [M+H] + .

[0089] 6e ((E)-4-(2-(4-(3-(4-Cyanophenyl)acryloyl)-3-methoxyphenoxy)ethoxy)-3-(phenylsulfonyl)-1,2,5-oxadiazole 2-oxide): Yield 19%, light yellow solid. 11H NMR (400 MHz, CDCl3) δ 7.98 (d, J = 8.6 Hz, 2H, 2”,6”-ArH), 7.75 (d, J = 8.6 Hz, 1H, 6-ArH), 7.74–7.69 (m, 1H, 4”-ArH), 7.66 (d, J = 15.7 Hz, 1H, Ar CH =CH-), 7.59–7.49 (m, 4H, 2’,6’,3”,5”-ArH), 7.39 (d, J = 15.8 Hz, 1H, ArCH= CH -), 6.92 (d, J = 8.6 Hz, 2H, 3’,5’-ArH), 6.59 (dd, J1 = 8.6 Hz, J2 = 2.2 Hz, 1H, 5-ArH), 6.54 (d, J = 2.1 Hz, 1H, 3-ArH), 4.66 (t, J = 6.0 Hz, 2H, -O CH 2CH2CH2O-), 4.26 (t, J = 5.8 Hz, 2H, -OCH2CH2 CH2 O-), 3.90 (s, 3H, 2-OCH3), 3.85 (s, 3H, 4’-OCH3), 2.40 (p, J = 5.9 Hz, 2H, -OCH2 CH2 CH2O-). ESI-MS m / z 548.1 [M+H] + .

[0090] 6f((E)-4-(3-(4-(4-Cyanophenyl)acryloyl)-3-methoxyphenoxy)propoxy)-3-(phenylsulfonyl)-1,2,5-oxadiazole 2-oxide): Yield 28%, pale yellow solid. 1 1H NMR (600 MHz, CDCl3) δ 8.02–7.97 (m, 2H, 2”,6”-ArH), 7.81 (t, J = 8.6 Hz, 1H, 6-ArH), 7.73 (t, J = 7.5 Hz, 1H, 4”-ArH), 7.67 (m, 4H, 2’,6’,3”,5”-ArH), 7.65 (d, J = 15.8 Hz, 1H, Ar CH =CH-), 7.61 (d, J = 15.8 Hz, 1H, ArCH= CH -), 7.55 (t, J = 8.1 Hz, 2H, 3’,5’-ArH), 6.61 (dd, J1 = 8.7 Hz, J2 = 2.2 Hz, 1H, 5-ArH), 6.55 (d, J = 2.2 Hz, 1H, 3-ArH), 4.66 (t, J = 6.0 Hz, 2H, -O CH2 CH2CH2O-), 4.28 (t, J = 5.8 Hz, 2H, -OCH2CH2 CH2O-), 3.93 (s, 3H, -OCH3), 2.40 (p, J=5.9Hz, 2H, -OCH2 CH2 CH2O-). ESI-MS m / z 561.9 [M+H] + .

[0091] Example 6

[0092] Synthesis of Compounds 7a-c in General Formula II

[0093] Place compounds 4a-c (4 mmol, 1 eq.), 5% palladium on carbon (20 wt%), and methanol (10 mL) in a 50 mL three-necked flask. Use a pipette to add diphenyl sulfide (7 μL, 0.01 eq.) to the flask. Replace the gas in the flask with H2 from a balloon and stir at room temperature for 1 day. After monitoring the reaction by TLC (PE:EA = 2:1) until completion, filter off the palladium on carbon. After rotary evaporation under reduced pressure, a solid crude product can be obtained, which is purified by column chromatography (PE:EA = 2:1) to give products 7a-c.

[0094] 7a (1-(4-Hydroxy-2-methoxyphenyl)-3-phenylpropan-1-one): Yield 63%, white solid. 1 1H NMR (400 MHz, CDCl3) δ 7.79–7.72 (d, J=9.0 Hz, 1H, ArH), 7.33–7.15 (m, 5H, ArH), 6.49–6.42 (m, 2H, ArH), 3.84 (s, 3H, -OCH3), 3.28 (t, J=7.7 Hz, 2H), 3.01 (t, J=7.7 Hz, 2H)

[0095] 7b (3-(4-Fluorophenyl)-1-(4-hydroxy-2-methoxyphenyl)propan-1-one): Yield 78%, white solid. 1 1H NMR (400 MHz, CDCl3) δ 7.73 (d, J=9.2 Hz, 1H), 7.17 (t, J=8.7 Hz, 2H), 6.96 (t, J=8.7 Hz, 2H), 6.49–6.42 (m, 2H), 3.84 (s, 3H), 3.25 (t, J=7.7 Hz, 2H), 2.98 (t, J=7.8 Hz, 2H)

[0096] 7c (1-(4-Hydroxy-2-methoxyphenyl)-3-(4-methoxyphenyl)propan-1-one): Yield 68%, white solid. 11H NMR (400 MHz, CDCl3) δ 7.73 (d, J = 9.2 Hz, 1H), 7.15 (d, J = 8.4 Hz, 2H), 6.83 (d, J = 8.6 Hz, 2H), 6.47–6.42 (m, 2H), 3.83 (s, 3H), 3.78 (s, 3H), 3.24 (t, J = 7.7 Hz, 2H), 2.94 (t, J = 7.8 Hz, 2H).

[0097] Example 7

[0098] Synthesis of Compounds 8a-f in General Formula II

[0099] The feeding ratio, reaction conditions, steps and operations of this example are the same as those for the synthesis of Compounds 5a-f in Example 4. The products 8a-f are directly subjected to the next reaction without purification.

[0100] Example 8

[0101] Synthesis of Compounds 9a-f in General Formula II

[0102] Place Compounds 8a-f (1 mmol, 1 eq.) and phenylsulfonylfurazan N-oxide (1.3 mmol, 1.3 eq.) in a reaction flask. Put the reaction flask in an ice-water bath, seal it with a rubber stopper, add anhydrous dichloromethane (10 mL). After 10 min, slowly add DBU (2 mmol, 2 eq.) and finish adding it in 30 min. Then stir for another 15 min and remove the ice bath. React at room temperature overnight. After 1 day, use LC-MS to detect that the reaction is basically complete. Extract three times with dichloromethane (15 mL), dry with anhydrous sodium sulfate, concentrate under reduced pressure by rotary evaporation to obtain a crude solid product. After purification by column chromatography (PE:DCM = 1:3), the target compounds 9a-f are obtained.

[0103] 9a (4-(2-(3-Methoxy-4-(3-phenylpropanoyl)phenoxy)ethoxy)-3-(phenylsulfonyl)-1,2,5-oxadiazole 2-oxide): Yield 26%, white solid. 1 1H NMR (400 MHz, CDCl3) δ 8.01 (d, J = 8.6 Hz, 2H, 2”,6”-ArH), 7.85 (d, J = 8.7 Hz, 1H, 6-ArH), 7.71 (t, J = 7.5 Hz, 1H, 4”-ArH), 7.53 (t, J = 7.6 Hz, 2H, 3”,5”-ArH), 7.34–7.16 (m, 5H, -ArH), 6.56 (dd, J1 = 8.7 Hz, J2 = 2.3 Hz, 1H, 5-ArH), 6.52 (d, J = 2.4 Hz, 1H, 3-ArH), 4.79 (t, J = 4.5 Hz, 2H, -O CH2CH2O-), 4.44 (t, J = 4.4 Hz, 2H, -OCH2 CH2 O-), 3.89 (s, 3H, -OCH3), 3.30 (t, J = 7.6 Hz, 2H, ArCH2 CH2 -), 3.02 (t, J = 8.0 Hz, 2H, Ar CH2 CH2-). HRMS(ESI) m / z: calcd for C 26 H 24 N2O8S + [M + H] + 525.1326, found 525.1327。

[0104] 9b (4-(3-(3-Methoxy-4-(3-phenylpropanoyl)phenoxy)propoxy)-3-(phenylsulfonyl)-1,2,5-oxadiazole 2-oxide): Yield 25%, white solid. 1 H NMR (400 MHz, CDCl3) δ 8.01 (d, J = 7.8 Hz, 2H), 7.83 (d, J = 8.7 Hz, 1H), 7.71 (t, J = 7.6 Hz, 1H), 7.54 (t, J = 7.9 Hz, 2H), 7.23–7.15 (m, 2H), 6.96 (t, J = 8.7 Hz, 2H), 6.56 (dd, J = 8.7, 2.3 Hz, 1H), 6.51 (d, J = 2.3 Hz, 1H), 4.79 (t, J = 4.3 Hz, 2H), 4.44 (t, J = 4.7 Hz, 2H), 3.90 (s, 3H), 3.26 (t, J = 7.6 Hz, 2H), 2.98 (t, J = 7.6 Hz, 2H). ESI-MS m / z 539.3 [M + H] + 。

[0105] 9c (4-(3-(3-(4-(4-Fluorophenyl)propanoyl)-3-methoxyphenoxy)propoxy)-3-(phenylsulfonyl)-1,2,5-oxadiazole 2-oxide): Yield 16%, white solid. 11H NMR (400 MHz, CDCl3) δ 8.00 (d, J = 7.8 Hz, 2H, 2”,6”-ArH), 7.83 (d, J = 8.7 Hz, 1H, 6-ArH), 7.71 (t, J = 7.6 Hz, 1H, 4”-ArH), 7.54 (t, J = 7.9 Hz, 2H, 3”,5”-ArH), 7.19 (dd, J1 = 8.5 Hz, J2 = 5.5 Hz, 2H, 2’,6’-ArH), 6.96 (t, J = 8.7 Hz, 2H, 3’,5’-ArH), 6.56 (dd, J1 = 8.7 Hz, J2 = 2.3 Hz, 1H, 5-ArH), 6.51 (d, J = 2.3 Hz, 1H, 3-ArH), 4.79 (t, J = 4.3 Hz, 2H, -O CH2 CH2O-), 4.44 (t, J = 4.7 Hz, 2H, -OCH2 CH2 O-), 3.90 (s, 3H, -OCH3), 3.26 (t, J = 7.6 Hz, 2H, ArCH2 CH2 -), 2.98 (t, J = 7.6 Hz, 2H, Ar CH2 CH2-). ESI-MS m / z 543.0 [M+H] + .

[0106] 9d (4-(3-(3-(4-(4-Fluorophenyl)propanoyl)-3-methoxyphenoxy)propoxy)-3-(phenylsulfonyl)-1,2,5-oxadiazole 2-oxide): Yield 23%, white solid. 1 1H NMR (400 MHz, CDCl3) δ 8.01 (d, J = 7.2 Hz, 2H, 2”,6”-ArH), 7.84 (d, J = 8.7 Hz, 1H, 6-ArH), 7.72 (t, J = 7.6 Hz, 1H, 4”-ArH), 7.54 (t, J = 7.9 Hz, 2H, 3”,5”-ArH), 7.19 (dd, J1 = 8.5 Hz, J2 = 5.5 Hz, 2H, 2’,6’-ArH), 6.97 (t, J = 8.7 Hz, 2H, 3’,5’-ArH), 6.57 (dd, J1 = 8.7 Hz, J2 = 2.3 Hz, 1H, 5-ArH), 6.52 (d, J = 2.3 Hz, 1H, 3-ArH), 4.80 (t, J = 4.4 Hz, 2H, -O CH2 CH2CH2O-), 4.45 (t, J = 4.9, 4.4 Hz, 2H), 3.90 (s, 3H, -OCH3), 3.27 (t, J = 7.6 Hz, 2H, ArCH2 CH2 -), 2.99 (t, J = 7.6 Hz, 2H, Ar CH2CH2-). ESI-MS m / z 557.2 [M+H] + .

[0107] 9e(4-(2-(3-methoxy-4-(3-(4-methoxyphenyl)propanoyl)phenoxy)ethoxy)-3-(phenylsulfonyl)-1,2,5-oxadiazole 2-oxide): Yield 32%, white solid. 1 H NMR (400 MHz, CDCl3) δ 8.01 (d, J = 7.6 Hz, 2H, 2”,6”-ArH), 7.83 (d, J = 8.6 Hz, 1H, 6-ArH), 7.71 (t, J = 7.5 Hz, 1H, 4”-ArH), 7.53 (t, J = 7.5 Hz, 2H, 3”,5”-ArH), 7.16 (d, J = 8.5 Hz, 2H, 2’,6’-ArH), 6.83 (d, J = 8.6 Hz, 2H, 3’,5’-ArH), 6.56 (dd, J1 = 8.7 Hz, J2 = 2.2 Hz, 1H, 5-ArH), 6.51 (d, J = 2.3 Hz, 1H, 3-ArH), 4.79 (t, J = 4.6 Hz, 2H, -O CH2 CH2O-), 4.44 (t, J = 4.5 Hz, 2H, -OCH2 CH2 O-), 3.89 (s, 3H, 2-OCH3), 3.79 (s, 3H, 4’-OCH3), 3.25 (t, J = 7.8 Hz, 2H, ArCH2 CH2 -), 2.95 (t, J = 7.8 Hz, 2H, Ar CH2 CH2-). ESI-MS m / z 555.3 [M+H] + .

[0108] 9f(4-(3-(3-methoxy-4-(3-(4-methoxyphenyl)propanoyl)phenoxy)propoxy)-3-(phenylsulfonyl)-1,2,5-oxadiazole 2-oxide): Yield 18%, white solid. 11H NMR (600 MHz, CDCl3) δ 7.98 (d, J = 8.4 Hz, 2H, 2”,6”-ArH), 7.82 (d, J = 8.7 Hz, 1H, 6-ArH), 7.71 (t, J = 7.5 Hz, 1H, 4”-ArH), 7.53 (t, J = 7.7 Hz, 2H, 3”,5”-ArH), 7.15 (d, J = 8.5 Hz, 2H, 2’,6’-ArH), 6.83 (d, J = 8.6 Hz, 2H, 3’,5’-ArH), 6.55 (dd, J1 = 8.7 Hz, J2 = 2.2 Hz, 1H, 5-ArH), 6.49 (d, J = 2.2 Hz, 1H, 3-ArH), 4.64 (t, J = 6.0 Hz, 2H, -O CH2 CH2CH2O-), 4.24 (t, J = 5.8 Hz, 2H, -OCH2CH2 CH2 O-), 3.87 (s, 3H, 2-OCH3), 3.78 (s, 3H, 4’-OCH3), 3.24 (t, J = 8.0 Hz, 2H, ArCH2 CH2 -), 2.95 (t, J = 7.8 Hz, 2H, Ar CH2 CH2-), 2.38 (p, J = 5.9 Hz, 2H, -OCH2 CH2 CH2O-). ESI-MS m / z 569.4 [M+H] + .

[0109] Example 9

[0110] Synthesis of compound 1-(4-hydroxy-2-methoxyphenyl)ethan-1-one (10) in general formula III

[0111] The crude product 3 (5 mmol) was diluted with 10 mL of methanol, 1 mL of 4 M dilute hydrochloric acid was added, and the mixture was stirred at room temperature for 12 h. TLC detected that the reaction was complete. It was extracted with water and ethyl acetate. The organic phase was dried over anhydrous sodium sulfate and concentrated by rotary evaporation under reduced pressure to obtain a crude solid. After recrystallization with ethanol, white solid 10 was obtained with a yield of 97.2%. The product did not need to be purified and was directly used as the raw material for the next reaction.

[0112] Example 10

[0113] Synthesis of compounds 11a-b in general formula III

[0114] The feeding ratio, reaction conditions, and operation steps of this reaction were the same as those for the synthesis of compounds 5a-f in Example 4. The products 11a-b were not purified and directly underwent the next reaction. Part of the products were purified by column chromatography (PE:EA = 2:1) to verify the structure, and the data are as follows.

[0115] 11a (1-(4-(2-hydroxyethoxy)-2-methoxyphenyl)ethan-1-one): 1 H NMR (400 MHz, CDCl3) δ 7.82 (d, J = 8.6 Hz, 1H, 6-ArH), 6.56–6.48 (m, 2H, 3,5-ArH), 4.14 (t, J = 4.5 Hz, 2H, -O CH2 CH2O-), 3.99 (t, J = 4.5 Hz, 2H, -OCH2 CH2 O-), 3.89 (s, 3H, -OCH3), 2.58 (s, 3H, -COCH3); ESI-MS m / z 210.9 [M+H] + 。

[0116] 11b (1-(4-(3-hydroxypropoxy)-2-methoxyphenyl)ethan-1-one): 1 H NMR (400 MHz, CDCl3) δ 7.82 (d, J = 8.8 Hz, 1H, 6-ArH), 6.53 (dd, J1 = 8.7 Hz, J2 = 2.3 Hz, 1H, 5-ArH), 6.46 (d, J = 2.3 Hz, 1H, 3-ArH), 4.18 (t, J = 6.1 Hz, 2H, -O CH2 CH2CH2O-), 3.89 (s, 3H, -OCH3), 3.87 (t, J = 5.8 Hz, 2H, -OCH2CH2 CH2 O-), 2.57 (s, 3H, -COCH3), 2.06 (m, 2H, -CH2-); ESI-MS m / z 225.0 [M+H] + 。

[0117] Example 11

[0118] Synthesis of Compounds 12a-b in General Formula III

[0119] The feeding ratio, reaction conditions, and operation steps were the same as those for the synthesis of Compounds 6a-f in Example 5. The product was purified by column chromatography (DCM:PE = 3:1) to obtain the target compounds 12a-b.

[0120] 12a (4-(2-(4-acetyl-3-methoxyphenoxy)ethoxy)-3-(phenylsulfonyl)-1,2,5-oxadiazole 2-oxide): Yield 37.9%, white solid. 11H NMR (400 MHz, CDCl3) δ 8.01 (d, J = 7.3 Hz, 2H, 2”,6”-ArH), 7.86 (d, J = 8.6 Hz, 1H, 6-ArH), 7.71 (t, J = 7.5 Hz, 1H, 4”-ArH), 7.54 (t, J = 8.1 Hz, 2H, 3”,5”-ArH), 6.55 (dd, J1 = 8.6 Hz, J2 = 2.3 Hz, 1H, 5-ArH), 6.52 (d, J = 2.2 Hz, 1H, 3-ArH), 4.79 (t, J = 4.5 Hz, 2H, -O CH2 CH2O-), 4.44 (t, J = 4.5 Hz, 2H, -OCH2 CH2 O-), 3.92 (s, 3H, -OCH3), 2.59 (s, 3H, -COCH3). ESI-MS m / z 435.2 [M+H] + .

[0121] 12b (4-(3-(4-acetyl-3-methoxyphenoxy)propoxy)-3-(phenylsulfonyl)-1,2,5-oxadiazole 2-oxide): Yield 44%, white solid. 1 1H NMR (400 MHz, CDCl3) δ 7.98 (d, J = 7.9 Hz, 2H, 2”,6”-ArH), 7.85 (d, J = 8.7 Hz, 1H, 6-ArH), 7.72 (t, J = 7.5 Hz, 1H, 4”-ArH), 7.53 (t, J = 8.0 Hz, 2H, 3”,5”-ArH), 6.55 (dd, J1 = 8.7 Hz, J2 = 2.3 Hz, 1H, 5-ArH), 6.50 (d, J = 2.2 Hz, 1H, 3-ArH), 4.64 (t, J = 6.0 Hz, 2H, -O CH2 CH2CH2O-), 4.24 (t, J = 5.8 Hz, 2H, -OCH2CH2 CH2 O-), 3.90 (s, 3H, -OCH3), 2.58 (s, 3H, -COCH3), 2.38 (p, J = 5.9 Hz, 2H, -OCH2 CH2 CH2O-). ESI-MS m / z 449.7 [M+H] + .

[0122] Example 12

[0123] Synthesis of compound 2-hydroxy-4-(2-hydroxyethoxy)benzoic acid (14) in general formula IV

[0124] Dissolve the raw material 2,4-dihydroxybenzoic acid (3.08 g, 20 mmol, 1 eq.) in ethanol (20 mL) and water (10 mL), place it in a constant temperature water bath at room temperature, add sodium hydroxide (2.0 g, 50 mmol, 2.5 eq.) and sodium iodide (300 mg, 2 mmol, 0.1 eq.), stir to dissolve, add bromoethanol (3 g, 24 mmol, 1.2 eq.), heat to 60 °C and react overnight. Then add dilute hydrochloric acid until the pH is less than 3 to terminate the reaction. Rotate and evaporate under reduced pressure to remove most of the ethanol, filter to separate the solid, extract the filtrate with water and ethyl acetate, collect the organic phase, wash it three times with saturated brine, dry it with anhydrous sodium sulfate, and obtain the crude product after rotary evaporation under reduced pressure. After column chromatography (PE:EA:AcOH = 2:1:0.01), it is combined with the previously filtered solid, and recrystallized with ethanol to obtain white solid 14 with a yield of 75.4%. 1 H NMR (400 MHz, DMSO-d6) δ 11.49 (bs, 1H), 7.69 (d, J = 8.8 Hz, 1H), 6.50 (dd, J = 8.7, 2.4 Hz, 1H), 6.47 (d, J = 2.4 Hz, 1H), 4.03 (t, J = 4.9 Hz, 2H), 3.70 (t, J = 4.9 Hz, 2H); ESI-MS m / z 199.4 [M+H] + .

[0125] Example 13

[0126] Synthesis of Compounds 15a-e in General Formula IV

[0127] Place 14 (800 mg, 4 mmol, 1 eq.), N-hydroxysuccinimide (NHS, 467 mg, 4 mmol, 1 eq.) and N,N'-dicyclohexylcarbodiimide (DCC, 825 mg, 4 mmol, 1 eq.) in a 50 mL three-necked flask, add 1,4-dioxane (10 mL), protect with N2, stir at room temperature for 12 h, filter out the solid, add the corresponding substituted benzylamine (4 mmol, 1 eq.), sodium bicarbonate (340 mg, 4 mmol, 1 eq.) and water (4 mL) to the filtrate, and heat to 60 °C and stir. After 5 h, monitor the reaction by TLC (PE:EA = 1:1), and when the reaction is basically complete, extract with water and ethyl acetate, wash three times with saturated brine, dry with anhydrous sodium sulfate, and obtain the white crude product after rotary evaporation under reduced pressure. After column chromatography (PE:EA = 2:1), products 15a-e are obtained.

[0128] 15a (N-benzyl-2-hydroxy-4-(2-hydroxyethoxy)benzamide): Yield 46.4%, white solid. 1HNMR(400MHz,CDCl3)δ12.66(s,1H),7.41–7.29(m,5H),7.26(d,J=8.6Hz,1H),6.48(d,J=2.5Hz,1H),6.41(dd,J1=8.8Hz,J2=2.6Hz,1H),6.39(br,1H),4.62(d,J=5.6Hz,2H),4.10(t,J=4.6Hz,2H),3.98(t,J=4.6Hz,2H).ESI-MSm / z 288.0[M+H] + .

[0129] 15b (N-(4-Fluorobenzyl)-2-hydroxy-4-(2-hydroxyethoxy)benzamide): Yield 45.1%, white solid; 1 H NMR(400MHz,CDCl3)δ7.83(d,J=8.6Hz,2H),7.59(d,J=8.6Hz,2H),7.09(d,J=8.6Hz,2H),6.52–6.41(m,2H),6.39(br,1H),4.60(d,J=5.5Hz,2H),4.56(t,J=4.0Hz,2H),4.18(t,J=4.0Hz,2H).ESI-MSm / z 306.1[M+H] + .

[0130] 15c ((4-((2-Hydroxy-4-(2-hydroxyethoxy)benzamido)methyl)phenyl)carbamic acid tert-butyl ester): Yield 48.5%, yellow solid; 1 H NMR(400MHz,CDCl3)δ7.34(d,J=8.1Hz,2H),7.23(d,J=8.3Hz,2H),6.54–6.34(m,4H),4.54(d,J=5.4Hz,2H),4.07(t,J=4.7Hz,2H),3.96(t,J=4.5Hz,2H),1.51(s,9H).ESI-MSm / z 403.1[M+H] + .

[0131] 15d (2-Hydroxy-4-(2-hydroxyethoxy)-N-(4-(trifluoromethyl)benzyl)benzamide): Yield 40.8%, white solid; 11H NMR (400 MHz, CDCl3) δ 8.21 (d, J = 8.8 Hz, 1H), 7.59 (d, J = 8.3 Hz, 2H), 7.46 (d, J = 8.2 Hz, 2H), 6.62 (dd, J1 = 8.8 Hz, J2 = 2.3 Hz, 1H), 6.54 (d, J = 2.3 Hz, 1H), 4.72 (d, J = 5.9 Hz, 2H), 4.14 (t, J = 4.3 Hz, 1H), 3.99 (t, J = 4.3 Hz, 1H), 3.92 (s, 3H). ESI-MS m / z 356.1 [M+H] + .

[0132] 15e (N-(4-Cyanobenzyl)-2-hydroxy-4-(2-hydroxyethoxy)benzamide): Yield 52.1%, white solid; 1 1H NMR (400 MHz, CDCl3) δ 8.26 (d, J = 8.5 Hz, 1H), 7.62 (d, J = 8.3 Hz, 2H), 7.48 (d, J = 8.2 Hz, 2H), 6.62 (dd, J1 = 8.7 Hz, J2 = 2.3 Hz, 1H), 6.54 (d, J = 2.3 Hz, 1H), 4.72 (d, J = 5.9 Hz, 2H), 4.14 (t, J = 4.1 Hz, 1H), 3.99 (t, J = 4.0 Hz, 1H), 3.92 (s, 3H). ESI-MS m / z 311.0 [M-H] - .

[0133] Example 14

[0134] Synthesis of Compounds 16a-f in General Formula IV

[0135] The feeding ratio, reaction conditions and operation steps are the same as those for the synthesis of Compounds 6a-f in Example 5. The products were purified by column chromatography (PE:EA = 2:1) to obtain the target compounds 16a-b, 16d-f. Among them, the intermediate 16c' was not purified. Trifluoroacetic acid (TFA, 5 mL) was added to the reaction solution and stirred for 30 min. After purification by column chromatography (PE:EA = 2:1), the target compound 16c was obtained.

[0136] 16a (4-(2-(4-(Benzylcarbamoyl)-3-hydroxyphenoxy)ethoxy)-3-(phenylsulfonyl)-1,2,5-oxadiazole 2-oxide): Yield 24.3%, white solid. 11H NMR (400 MHz, CDCl3) δ 8.01 (d, J = 7.7 Hz, 2H, 2”,6”-ArH), 7.71 (t, J = 7.4 Hz, 1H, 4”-ArH), 7.54 (t, J = 7.4 Hz, 2H, 3”,5”-ArH), 7.41–7.29 (m, 6H, ArH), 6.49 (d, J = 2.5 Hz, 1H, 3-ArH), 6.44–6.39 (m, 2H, 5-ArH, -CONH-), 4.76 (t, J = 4.4 Hz, 2H, -O CH2 CH2O-), 4.64 (d, J = 5.5 Hz, 2H, Ar CH2 NH-), 4.38 (t, J = 4.4 Hz, 2H, -OCH2 CH2 O-). ESI-MS m / z 512.4 [M+H] + .

[0137] 16b (4-(2-(4-((4-Fluorobenzyl)carbamoyl)-3-hydroxyphenoxy)ethoxy)-3-(phenylsulfonyl)-1,2,5-oxadiazole 2-oxide): Yield 22.2%, white solid. 1 1H NMR (400 MHz, CDCl3) δ 8.01 (d, J = 7.8 Hz, 2H, 2”,6”-ArH), 7.73 (t, J = 7.6 Hz, 1H, 4”-ArH), 7.53 (t, J = 7.4 Hz, 2H, 3”,5”-ArH), 7.37–7.29 (m, 3H, 6,3’,5’-ArH), 7.05 (t, J = 8.6 Hz, 2H, 2’,6’-ArH), 6.49 (d, J = 1.9 Hz, 1H, 3-ArH), 6.47–6.41 (m, 2H, 5-ArH, -CONH-), 4.76 (t, J = 3.6 Hz, 2H, -O CH2 CH2O-), 4.60 (d, J = 5.5 Hz, 2H, Ar CH2 NH-), 4.38 (d, J = 3.8 Hz, 2H, -OCH2 CH2 O-). ESI-MS m / z 530.1 [M+H] + .

[0138] 16c (4-(2-(4-((4-Aminobenzyl)carbamoyl)-3-hydroxyphenoxy)ethoxy)-3-(phenylsulfonyl)-1,2,5-oxadiazole 2-oxide): Yield 28.6%, white solid. 11H NMR (400 MHz, DMSO-d6) δ 13.18 (s, 1H, -ArOH), 9.06 (t, J = 5.8 Hz, 1H, -CONH-), 7.97 (d, J = 7.8 Hz, 2H, 2”,6”-ArH), 7.90–7.80 (m, 2H, 6,4”-ArH), 7.66 (t, J = 7.7 Hz, 2H, 3”,5”-ArH), 6.99 (d, J = 8.5 Hz, 2H, 2’,6’-ArH), 6.55–6.46 (m, 4H, 3,5,3’,5’-ArH), 4.99 (s, 2H, -NH2), 4.72 (t, J = 4.5 Hz, 2H, -O CH2 CH2O-), 4.40 (d, J = 4.2 Hz, 2H, -OCH2 CH2 O-), 4.30 (d, J = 5.6 Hz, 2H, Ar CH2 NH-). ESI-MS m / z 527.0 [M+H] + .

[0139] 16d (4-(2-(3-Hydroxy-4-((4-(trifluoromethyl)benzyl)carbamoyl)phenoxy)ethoxy)-3-(phenylsulfonyl)-1,2,5-oxadiazole 2-oxide): Yield 28.7%, white solid. 1 1H NMR (400 MHz, DMSO-d6) δ 12.87 (s, 1H, -ArOH), 9.32 (t, J = 6.0 Hz, 1H, -CONH-), 7.98 (d, J = 7.8 Hz, 2H, 2”,6”-ArH), 7.92–7.80 (m, 2H, 6,4”-ArH), 7.74–7.62 (m, 4H, 3’,5’,3”,5”-ArH), 7.54 (d, J = 8.0 Hz, 2H, 2’,6’-ArH), 6.58 (dd, J1 = 8.7 Hz, J2 = 2.2 Hz, 1H, 5-ArH), 6.52 (d, J = 2.4 Hz, 1H, 3-ArH), 4.73 (t, J = 3.4 Hz, 2H, -O CH2 CH2O-), 4.58 (d, J = 5.8 Hz, 2H, Ar CH2 NH-), 4.41 (t, J = 3.6 Hz, 2H, -OCH2 CH2 O-). ESI-MS m / z 580.2 [M+H] + .

[0140] 16e (4-(2-(4-((4-Cyanobenzyl)carbamoyl)-3-hydroxyphenoxy)ethoxy)-3-(phenylsulfonyl)-1,2,5-oxadiazole 2-oxide): Yield 18.7%, light yellow solid.1 1H NMR (600 MHz, DMSO-d6) δ 12.79 (s, 1H, -ArOH), 9.29 (t, J = 5.9 Hz, 1H, -CONH-), 7.97 (d, J = 8.0 Hz, 2H, 2”,6”-ArH), 7.87 (d, J = 8.9 Hz, 1H, 6-ArH), 7.84 (t, J = 7.4 Hz, 1H, 4”-ArH), 7.81 (d, J = 8.3 Hz, 2H, 3’,5’-ArH), 7.66 (t, J = 8.0 Hz, 2H, 3”,5”-ArH), 7.51 (d, J = 8.3 Hz, 2H, 2’,6’-ArH), 6.57 (dd, J1 = 8.9 Hz, J2 = 2.6 Hz, 1H, 5-ArH), 6.50 (d, J = 2.6 Hz, 1H, 3-ArH), 4.73 (t, J = 4.0 Hz, 2H, -O CH2 CH2O-), 4.57 (d, J = 5.8 Hz, 2H, Ar CH2 NH-), 4.40 (t, J = 4.0 Hz, 2H, -OCH2 CH2 O-). ESI-MS m / z 537.3 [M+H] + .

[0141] 16f (4-(2-(3-Hydroxy-4-((4-hydroxybenzyl)carbamoyl)phenoxy)ethoxy)-3-(phenylsulfonyl)-1,2,5-oxadiazole 2-oxide): Yield 16.5%, white solid; 1 1H NMR (400 MHz, DMSO-d6) δ 12.95 (s, 1H), 9.29 (s, 1H), 8.03 (d, J = 7.8 Hz, 2H), 7.91–7.80 (m, 3H), 7.65 (t, J = 8.0 Hz, 2H), 7.43 (d, J = 7.9 Hz, 2H), 7.39 (d, J = 8.1 Hz, 2H), 6.58 (dd, J = 8.8, 2.5 Hz, 1H), 6.51 (d, J = 2.5 Hz, 1H), 4.72 (t, J = 4.5 Hz, 2H), 4.52 (d, J = 5.1 Hz, 2H), 4.40 (t, J = 4.5 Hz, 2H). ESI-MS m / z 528.3 [M+H] + .

[0142] Example 15

[0143] Synthesis of Compounds 17a-b and 17d-e in General Formula V

[0144] The feeding ratio, reaction conditions, and operation steps are the same as those for the synthesis of Compound 3 in Example 2, with the reaction temperature adjusted to room temperature. Products 17a-b and 17d-e can be directly subjected to the next reaction without purification. Part of the crude product was taken out and purified by column chromatography (PE:EA = 2:1) to verify the structure.

[0145] 17a (N-Benzyl-4-(2-hydroxyethoxy)-2-methoxybenzamide): 1 H NMR (400 MHz, CDCl3) δ 8.23 (d, J = 8.9 Hz, 1H, 6-ArH), 8.12 (br, 1H, -CONH-), 7.39–7.28 (m, 5H, ArH), 6.62 (dd, J1 = 8.8 Hz, J2 = 2.5 Hz, 1H, 5-ArH), 6.54 (d, J = 2.2 Hz, 1H, 3-ArH), 4.69 (d, J = 5.1 Hz, 2H, Ar CH2 NH-), 4.15 (t, J = 4.2 Hz, 2H, -O CH2 CH2O-), 4.00 (t, J = 4.2 Hz, 2H, -OCH2 CH2 O-), 3.90 (s, 3H, -OCH3).

[0146] 17b (N-(4-Fluorobenzyl)-4-(2-hydroxyethoxy)-2-methoxybenzamide): 1 H NMR (400 MHz, CDCl3) δ 8.20 (d, J = 8.7 Hz, 1H, 6-ArH), 8.08 (br, 1H, -CONH-), 7.32 (t, J = 8.8 Hz, 2H, 2’,6’-ArH), 7.02 (t, J = 8.9 Hz, 2H, 3’,5’-ArH), 6.61 (dd, J1 = 8.8 Hz, J2 = 2.4 Hz, 1H, 5-ArH), 6.53 (d, J = 2.3 Hz, 1H, 3-ArH), 4.62 (d, J = 5.7 Hz, 2H, Ar CH2 NH-), 4.13 (t, J = 3.9 Hz, 2H, -O CH2 CH2O-), 3.98 (t, J = 3.9 Hz, 2H, -OCH2 CH2 O-), 3.89 (s, 3H, -OCH3).

[0147] 17d (4-(2-Hydroxyethoxy)-2-methoxy-N-(4-(trifluoromethyl)benzyl)benzamide): 11H NMR (400 MHz, CDCl3) δ 8.21 (d, J = 8.8 Hz, 1H, 6 - ArH), 7.59 (d, J = 8.3 Hz, 2H, 3’,5’ - ArH), 7.46 (d, J = 8.2 Hz, 2H, 2’,6’ - ArH), 6.62 (dd, J1 = 8.8 Hz, J2 = 2.3 Hz, 1H, 5 - ArH), 6.54 (d, J = 2.3 Hz, 1H, 3 - ArH), 4.72 (d, J = 5.9 Hz, 2H, Ar CH2 NH - ), 4.14 (t, J = 4.3 Hz, 2H, - O CH2 CH2O - ), 3.99 (t, J = 4.3 Hz, 2H, - OCH2 CH2 O - ), 3.92 (s, 3H, - OCH3).

[0148] 17e (N - (4 - cyanobenzyl) - 4 - (2 - hydroxyethoxy) - 2 - methoxybenzamide): 1 1H NMR (400 MHz, CDCl3) δ 8.23–8.16 (m, 2H), 7.62 (d, J = 7.2 Hz, 1H), 7.45 (d, J = 7.5 Hz, 1H), 6.62 (dd, J = 8.8, 2.1 Hz, 1H), 6.55 (d, J = 2.1 Hz, 1H), 4.71 (d, J = 5.9 Hz, 2H), 4.14 (t, J = 4.8 Hz, 2H), 3.99 (t, J = 4.8 Hz, 2H), 3.93 (s, 3H).

[0149] Example 16

[0150] Synthesis of Compounds 18a - b and 18d - e in General Formula V

[0151] The feeding ratio, reaction conditions, and operation steps are the same as those for the synthesis of Compounds 6a - f in Example 5. The target compounds 18a - b and 18d - e can be obtained by purification through column chromatography (PE:EA = 2:1).

[0152] 18a (4 - (2 - (4 - (benzylcarbamoyl) - 3 - methoxyphenoxy)ethoxy) - 3 - (phenylsulfonyl) - 1,2,5 - oxadiazole 2 - oxide): Yield 17%, white solid. 11H NMR (400 MHz, CDCl3) δ 8.27 (d, J = 8.6 Hz, 1H, 6-ArH), 8.11 (t, J = 5.0 Hz, 1H, -CONH-), 8.00 (d, J = 7.5 Hz, 2H, 2”,6”-ArH), 7.72 (t, J = 7.4 Hz, 1H, 4”-ArH), 7.52 (t, J = 7.6 Hz, 2H, 3”,5”-ArH), 7.38–7.27 (m, 5H, -ArH), 6.64 (dd, J1 = 8.8 Hz, J2 = 2.3 Hz, 1H, 5-ArH), 6.55 (d, J = 2.2 Hz, 1H, 3-ArH), 4.79 (d, J = 3.6 Hz, 2H, -O CH2 CH2O-), 4.69 (d, J = 5.3 Hz, 2H, Ar CH2 NH-), 4.44 (t, J = 3.4 Hz, 2H, -OCH2 CH2 O-), 3.92 (s, 3H, -OCH3); ESI-MS m / z 526.2 [M+H] + .

[0153] 18b (4-(2-(4-((4-Fluorobenzyl)carbamoyl)-3-methoxyphenoxy)ethoxy)-3-(phenylsulfonyl)-1,2,5-oxadiazole 2-oxide): Yield 23%, white solid. 1 1H NMR (400 MHz, CDCl3) δ 8.26 (d, J = 8.8 Hz, 1H, 6-ArH), 8.09 (t, J = 6.5 Hz, 1H, -CONH-), 8.00 (d, J = 7.8 Hz, 2H, 2”,6”-ArH), 7.70 (t, J = 7.5 Hz, 1H, 4”-ArH), 7.53 (t, J = 7.5 Hz, 2H, 3”,5”-ArH), 7.33 (t, J = 6.8 Hz, 2H, 3’,5’-ArH), 7.02 (t, J = 8.8 Hz, 2H, 2’,6’-ArH), 6.64 (dd, J1 = 8.9 Hz, J2 = 2.2 Hz, 1H, 5-ArH), 6.56 (d, J = 2.4 Hz, 1H, 3-ArH), 4.79 (t, J = 4.5 Hz, 2H, -O CH2 CH2O-), 4.64 (d, J = 5.7 Hz, 2H, Ar CH2 NH-), 4.43 (t, J = 4.5 Hz, 2H, -OCH2 CH2 O-), 3.93 (s, 3H, -OCH3). ESI-MS m / z 544.1 [M+H] + .

[0154] 18d (4-(2-(3-Methoxy-4-((4-(trifluoromethyl)benzyl)carbamoyl)phenoxy)ethoxy)-3-(phenylsulfonyl)-1,2,5-oxadiazole 2-oxide): Yield 29%, white solid. 1 H NMR (400 MHz, DMSO-d6) δ 8.72 (t, J = 6.1 Hz, 1H, -CONH-), 7.98 (d, J = 7.9 Hz, 2H, 2”,6”-ArH), 7.88–7.80 (m, 2H, 6,4”-ArH), 7.72–7.62 (m, 4H, 3’,5’,3”,5”-ArH), 7.53 (d, J = 7.9 Hz, 2H, 2’,6’-ArH), 6.72 (d, J = 2.3 Hz, 1H, 3-ArH), 6.70 (dd, J1 = 8.5 Hz, J2 = 2.3 Hz, 1H, 5-ArH), 4.76 (t, J = 4.3 Hz, 2H, -O CH2 CH2O-), 4.58 (d, J = 6.1 Hz, 2H, Ar CH2 NH-), 4.46 (t, J = 4.1 Hz, 1H, -OCH2 CH2 O-), 3.94 (s, 3H, -OCH3). ESI-MS m / z 594.0 [M+H] + .

[0155] 18e (4-(2-(4-((4-Cyanobenzyl)carbamoyl)-3-methoxyphenoxy)ethoxy)-3-(phenylsulfonyl)-1,2,5-oxadiazole 2-oxide): Yield 15%, light yellow solid. 1 H NMR (400 MHz, DMSO-d6) δ 8.72 (t, J = 6.1 Hz, 1H, -CONH-), 7.98 (d, J = 7.9 Hz, 2H, 2”,6”-ArH), 7.88–7.74 (m, 4H, 6,3’,5’,4”-ArH), 7.67 (t, J = 7.9 Hz, 2H, 3”,5”-ArH), 7.50 (d, J = 8.0 Hz, 2H, 2’,6’-ArH), 6.72 (d, J = 2.3 Hz, 1H, 3-ArH), 6.70 (dd, J1 = 8.6 Hz, J2 = 2.3 Hz, 1H, 5-ArH), 4.76 (t, J = 4.1 Hz, 2H, -O CH2 CH2O-), 4.57 (d, J = 6.1 Hz, 2H, Ar CH2 NH-), 4.46 (t, J = 4.4 Hz, 2H, -OCH2 CH2 O-), 3.94 (s, 3H, -OCH3). ESI-MS m / z551.2 [M+H] +.

[0156] Example 17 Antitumor Activity Test at the Cellular Level

[0157] The MTS method was used to detect the activity of the compounds of the present invention in inhibiting the proliferation of tumor cells. Cancer cells in the logarithmic growth phase were inoculated into 96-well plates and divided into a blank control group, a positive control group, and treatment groups with different concentrations of the compounds of the present invention or the positive drug. After culturing at 37 °C for 48 h, MTT detection was carried out, and the cell proliferation inhibition rate and IC 50 . Most of the target compounds have comparable or higher selective proliferation inhibition activity (i.e., accessory sensitivity) against drug-resistant breast cancer MCF-7 / ADR cells than CY-16S-4A93, and the IC 50 is between 1.09 - 91.1 nM. Among them, compounds with an open-chain side chain of α,β-unsaturated ketone (6a-f) and α,β-saturated ketone (9a-f) all have excellent proliferation inhibition activity. Except for the IC 50 of compound 6c being 12.42 nM, the IC 50 of the remaining compounds are all at the nanomolar level. The activity of compounds 12a-b with the benzene ring removed from the side chain in inhibiting the proliferation of MCF-7 / ADR cells is 25 - 50 times lower than that of the corresponding α,β-unsaturated ketone compound 6a or α,β-saturated ketone compound 9a-b with a benzene ring in the side chain. After replacing the coumarin lactone ring with an amide side chain, the target compounds 16a-f with the 2-hydroxy group of the parent benzene ring exposed, and the target compounds 18a-b and 18d-e modified with methoxy groups also have strong proliferation inhibition activity against MCF-7 / ADR cells, and the IC 50 is 5.36 - 91.1 nM. In addition, four target compounds 9a-b and 9d-e with four side chains of α,β-saturated ketone show significant accessory sensitivity, and the IC 50(MCF-7) / IC 50(MCF-7 / ADR) ratio exceeds 1000 times. Compound 9f also reaches a selective inhibition of the proliferation activity of drug-resistant breast cancer MCF-7 / ADR cells of more than 940 times (see Table 1).

[0158] The compounds of the present invention show only weak cell proliferation inhibition effects on the proliferation inhibition activities of two triple-negative breast cancer cells, MDA-MB-231 and MDA-MB-468. Among them, the IC 50 values of most target compounds for MDA-MB-231 cells are between 1 - 6 μM. The proliferation inhibition activity against MDA-MB-468 cells increases, and most of the IC 50 are between 80 - 780 nM, and the activities are all inferior to the inhibition effect on the drug-resistant strain MCF-7 / ADR (see Table 2).

[0159] Most of the target compounds of the present invention have relatively low toxicity to normal cells MCF-10A, and the IC 50 is between 1.8 - 13.4 μM (see Table 3). The IC 50 values of all compounds for HUVEC are between 1.9 - 17.0 μM. Based on the toxicity evaluation results for MCF-10A, the cell viability of HUVEC was detected for 6a, 6d, 6e, 9a, 9e, and compound 12a at a dosing concentration of 500 nM, and it also showed good safety (see Figure 2 ).

[0160] Table 1 shows the anti-proliferative activities of the target compounds against MCF-7 and drug-resistant MCF-7 / ADR breast cancer cell lines

[0161] Table 2 shows the anti-proliferative activities of the target compounds against MDA-MB-231 and MDA-MB-468 triple-negative breast cancer cell lines. Table 3 shows the anti-proliferative activities of the active target compounds against normal breast cells MCF-10A

[0162] Table 4 shows the solubility ratio of the target compounds to CY-16S-4A93

[0163] Table 1

[0164]

[0165]

[0166] Table 2

[0167]

[0168]

[0169] Table 3

[0170]

[0171] Example 18 Determination of intracellular NO release

[0172] Cells in the logarithmic growth phase were seeded in 6-well plates at a density of 2×10 5 cells per well. After overnight incubation, the specified concentration of 9e was added to the cells. After incubation in a 37°C incubator for 4 hours, the original culture medium was aspirated. For total intracellular NO, the DAF-FM DA probe was diluted to 5 μM using a medium without fetal bovine serum and incubated at 37°C for 15 minutes. After washing the cells 3 times with PBS, Hoechst 33342 was diluted at a volume ratio of 1:1000 using a medium without fetal bovine serum and incubated at 37°C for 10 minutes. After staining, the cells were washed 3 times with PBS, and fluorescence images were taken using an inverted fluorescence microscope with blue nuclei.

[0173] For lysosome-specific NO, incubate with 10 μM Lyso-NO probe at 37 °C for 15 minutes. After rinsing the cells 3 times with PBS, incubate with Hoechst 33342 at 37 °C for 10 minutes. After staining, rinse the cells 3 times with PBS, and take fluorescence images with an inverted fluorescence microscope. The nucleus is blue, and lysosome-specific NO appears green. After taking pictures, use Image J software to analyze the average fluorescence intensity of the green fluorescence.

[0174] The results showed that after treating MCF-7 / ADR with 10 nM concentration of 9e for 1 hour, a significant increase in the green fluorescence intensity of the lysosome-specific NO fluorescence probe (Lyso-NO) was found, showing a significant difference from the control group. On the other hand, treating the breast cancer sensitive strain MCF-7 with the same concentration of the target compound 9e for the same time showed no significant difference in the NO level in the lysosomes of MCF-7 cells before and after treatment. It is indicated that the highly selective inhibitory activity of compound 9e against drug-resistant breast cancer cells may be related to its ability to selectively enter the lysosomes of MCF-7 / ADR and release NO (see Figure 3 ).

[0175] Example 19 Autophagy induction in drug-resistant breast cancer MCF-7 / ADR cells by active compound 9e

[0176] After co-incubating the target compound 9e at a concentration of 10 nM with MCF-7 / ADR for 0, 6, 12, and 24 h, immunofluorescence staining of the cells was performed simultaneously with LC3 and LAMP1 antibodies. The experimental results showed that at 6 h, there was no significant change in the fluorescence intensity of LC3 and LAMP1, but autophagosomes gradually showed good co-localization with lysosomes (at the white arrow), indicating that autophagolysosomes were generated in MCF-7 / ADR under the induction of compound 9e. Autophagosomes accumulated at 12 - 24 h; meanwhile, the fluorescence of LAMP1 decreased, indicating a decrease in lysosomal activity. It is inferred that the target compound 9e can induce the generation of LC3 autophagosomes in MCF-7 / ADR, but due to the dysfunction of lysosomes caused by the accumulation of NO, the damaged lysosomes cannot degrade the LC3-II protein on the inner membrane of autophagosomes, resulting in the accumulation of autophagosomes and a decrease in lysosomal activity (see Figure 4 ).

[0177] Example 20 Promotion of ferroptosis in MCF-7 / ADR cells by active compound 9e

[0178] MCF-7 / ADR was co-incubated with different concentrations of the ferroptosis inhibitor Fer-1, and then the inhibitory effect of compound 9e on cell proliferation was detected by MTT colorimetric assay. As the concentration of the Fer-1 reagent increased, the inhibitory rate of compound 9e on the proliferation of MCF-7 / ADR cells gradually decreased, showing a concentration-dependent relationship. When the cells were treated with 20 nM concentration of 9e alone, the proliferation inhibitory rate was 89%; while after pretreatment of MCF-7 / ADR with 200 nM concentration of Fer-1, the proliferation inhibitory rate of 9e was only 2%, and the inhibitory activity was completely lost. This indicates that there is a close relationship between the proliferation inhibitory activity of the target compound 9e and the induction of ferroptosis in cells (see Figure 5 ).

[0179] Example 21 Solubility ratio of the target compound to CY-16S-4A93

[0180] Using CY-16S-4A93 as a control, the solubility of the above-mentioned target compounds in a mixture of water:acetonitrile at 2:1 was tested by HPLC. The results showed that the solubility of these newly synthesized target compounds was increased by 24 - 775 times compared with CY-16S-4A93. Compound 16c, which contains an amino group on the benzene ring of the amide side chain, had the best solubility, with a 409-fold increase, indicating that the introduction of hydrophilic groups is beneficial to improving solubility. The solubility of the most active compound 9e was relatively weaker than that of the compounds with amide chains, but still 71 times higher than that of the control CY-16S-4A93. Therefore, the flexible side-chain skeleton aromatic ketone / furazan conjugates have better solubility than the coumarin skeleton / furazan compounds (as shown in Table 4).

[0181] Table 4

[0182]

[0183] The above-described embodiments are only descriptions of the preferred embodiments of the present invention and do not limit the scope of the present invention. Without departing from the spirit of the present invention, various modifications and improvements made by those of ordinary skill in the art to the technical solutions of the present invention shall fall within the protection scope determined by the claims of the present invention.

Claims

1. An aryl ketone derivative-furazan fused conjugate, characterized in that, The structural formula is as shown in any one of Formulas I to V: In Formulas I to V, R2, R3, R4, and R5 are each independently one of H, F, OCH3, CN, NH2, CF3, and OH; n = 2 or 3.

2. The conjugate of the aryl ketone derivative and furazan according to claim 1, wherein, In Formula I, R2 = H, F, OCH3 or CN; In Formula II, R3 = H, F or OCH3; In Formula IV, R4 = H, F, NH2, CF3, CN or OH; In Formula V, R5 = H, F, NH2, CF3 or CN.

3. A method for preparing a conjugate of an aryl ketone derivative and furazan, as claimed in claim 1 or 2, characterized in that when the structural formula of the conjugate of the aryl ketone derivative and furazan is as shown in Formula I, the method for preparing the conjugate of the aryl ketone derivative and furazan includes the following steps: Using 2,4-dihydroxyacetophenone as a raw material, protecting the 4-position hydroxyl group in the raw material with dihydropyran under the catalysis of 4-toluenesulfonic acid pyridine salt to obtain Compound 2; methylating Compound 2 with CH3I to generate Compound 3, and then performing an aldol condensation reaction with benzaldehydes containing different substituents, and removing the pyran protecting group under hydrochloric acid to synthesize intermediates 4a-d; In the presence of K2CO3 and NaI, intermediates 4a-d are etherified with 2-bromoethanol or 3-bromo-1-propanol to obtain Compounds 5a-f; At room temperature, in dichloromethane solution and under the catalytic condition of 1,8-diazabicyclo[5.4.0]-7-undecene, Compounds 5a-f are coupled with phenylsulfonyl furazan N-oxide to obtain a conjugate of an aryl ketone derivative and furazan with the structure shown in Formula I; when the structural formula of the conjugate of the aryl ketone derivative and furazan is as shown in Formula II, the method for preparing the conjugate of the aryl ketone derivative and furazan includes the following steps: The double bond in the α,β-unsaturated ketone side chain of intermediates 4a-d is partially reduced with palladium-carbon in phenyl sulfide / methanol to synthesize intermediates 7a-c, and then etherified with 2-bromoethanol or 3-bromo-1-propanol to obtain Compounds 8a-f; at room temperature, under the catalysis of DBU / DCM, Compounds 8a-f are coupled with phenylsulfonyl furazan N-oxide to obtain a conjugate of an aryl ketone derivative and furazan with the structure shown in Formula II; when the structural formula of the conjugate of the aryl ketone derivative and furazan is as shown in Formula III, the method for preparing the conjugate of the aryl ketone derivative and furazan includes the following steps: Removing the pyran protecting group from Compound 3 to obtain 4-hydroxy-2-methoxyacetophenone; 4-hydroxy-2-methoxyacetophenone is refluxed with bromoethanol or bromopropanol, potassium carbonate and sodium iodide in DMF to obtain 4-hydroxyethoxy- or hydroxypropoxy-substituted 2-methoxyacetophenone, and finally reacting with phenylsulfonyl furazan N-oxide to obtain a conjugate of an aryl ketone derivative and furazan with the structure shown in Formula III; when the structural formula of the conjugate of the aryl ketone derivative and furazan is as shown in Formula IV, the method for preparing the conjugate of the aryl ketone derivative and furazan includes the following steps: In ethanol, compound 13 reacts with bromoethanol, sodium hydroxide and sodium iodide under stirring to obtain 4-hydroxyethoxysalicylic acid; 4-hydroxyethoxysalicylic acid condenses with substituted ampicillin to form amide side chain intermediates 15a-f, and then reacts with phenylsulfonylfurazan N-oxide in DCM containing DBU at room temperature to synthesize the conjugate of the aryl ketone derivative shown in formula IV and furazan. The structural formula of the said Compound 13 is When the conjugate of the aryl ketone derivative and furazan has the structural formula shown in formula V, the preparation method of the conjugate of the aryl ketone derivative and furazan includes the following steps: Intermediates 15a-b and 15d-e are first etherified with CH3I under the catalysis of potassium carbonate to form 2-methoxy derivatives, and then react with phenylsulfonylfurazan N-oxide in DCM containing DBU at room temperature to synthesize the conjugate of the aryl ketone derivative shown in formula V and furazan.

4. Use of the conjugate of the aryl ketone derivative and furazan according to claim 1 or 2 in the preparation of anti-tumor drugs.

5. The application according to claim 3, characterized in that The tumor is caused by human triple-negative breast cancer cells MDA-MB-231 and MDA-MB-468, breast cancer cells MCF-7 or drug-resistant tumor cells MCF-7 / ADR.

6. An antitumor drug, characterized in that, The active ingredient includes the conjugate of the aryl ketone derivative and furazan according to claim 1 or 2.

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