2-triazole ethane-1-ketone / alcohol class of hsp90 / hif-1 inhibitors and uses

Abstract

The application discloses a kind of 2-triazole ethane-1-ketone / alcohol HSP90 / HIF-1 inhibitor and application, new drug use technical field.This kind of inhibitor is transformed from 7-hydroxy new piece helicon A, can low toxicity efficiently inhibit the expression of HIF-1, inhibit tumor cell invasion, clonal proliferation and other malignant development process, in vivo anti-tumor activity experiment and cisplatin can effectively inhibit tumor.This kind of inhibitor can be applied to the preparation of inhibiting cancer development drug.

Description

TECHNICAL FIELD

[0001] The present application relates to the technical field of new use of drugs, in particular to a class of 2-triazole ethane-1-ketone / alcohol HSP90 / HIF-1 and application BACKGROUND

[0002] Hypoxia is a feature widely existing in solid tumors, and hypoxic conditions induce tumor cells to produce hypoxia-inducible factor-1 (HIF-1). HIF-1 is an essential tumor cell survival factor, which is composed of two subunits of HIF-1α and HIF-1β. The activity of HIF-1 is mainly determined by the stability of HIF-1α. HIF-1α is extremely unstable under normoxic conditions and is rapidly degraded under the mediation of the tumor suppressor VHL, but is stable under hypoxic conditions due to the inability of VHL to bind to HIF-1. It is then translocated into the nucleus, and with the assistance of coactivators, it heterodimerizes with HIF-1β, activates the transcription of a variety of genes related to tumor occurrence and development, helps tumor cells adapt to hypoxic environment, and inhibiting HIF-1 activity has become an effective and attractive strategy for treating various solid tumors.

[0003] Heat shock protein 90 (HSP90) is widely present in cells, but is highly expressed in tumor cells and exists in a complex complex form different from that in normal cells. HSP90 has numerous client proteins, of which more than 40 are currently known, including Akt, human epidermal growth factor (Her2 / ErbB2), Bcr-Abl, Raf-1, HIF-1α, Her2, etc. HSP90 is an important regulator of HIF-1α protein stability. Studies have shown that inhibitors of HSP90 can promote the degradation of HIF-1α in different ways, and the activity of HIF-1 is directly and closely related to the activity of HSP90.

[0004] HIF-1 and HSP90 are important for the development of cancer inhibitors of the HSP90 / HIF-1 pathway by inhibiting cancer metastasis and other malignant development processes, which can be used for the development of new cancer treatment methods. SUMMARY

[0005] The present application aims to modify the structure of 7-hydroxyneopeltaxonin A, overcome the problems of long synthesis route and low yield of 7-hydroxyneopeltaxonin A, design and synthesize a class of 2-triazole ethane-1-ketone / alcohol derivatives, which have low toxicity, can effectively inhibit tumor cell invasion, clonal proliferation and malignant development process, and develop anti-tumor development drugs. The drugs can be used together with cytotoxic drugs to reduce the dosage of cytotoxic drugs and achieve better inhibition effect on tumors.

[0006]

[0007] To achieve the object of the present application, the present application provides the following technical solutions.

[0008] The compound of formula I:

[0009]

[0010] wherein R is or

[0011] R1 is phenyl or phenyl containing at least one substituent, and n is an integer from 0 to 5.

[0012] R2 is phenyl or phenyl containing at least one substituent.

[0013] each of the substituents is independently selected from the group consisting of hydroxyl, carboxyl, amino, C1-C10 alkylamino, mercapto, C1-C10 thioether, C1-C10 alkoxy, -COOR0, -CONR0, halogen, nitro, cyano, aldehyde, C1-C10 alkyl, C1-C10 substituted alkyl, C1-C10 cycloalkyl, C1-C5 fluoroalkyl, C1-C10 alkenyl, adamantyl, m-carboranyl, o-carboranyl, C1-C13 heterocyclic group, C1-C13 substituted heterocyclic group;

[0014] the substituents of the substituted heterocyclic group are selected from at least one of the group consisting of hydroxyl, carboxyl, amino, C1-C10 alkylamino, mercapto, C1-C10 alkoxy, -COOR0, -CONR0, halogen, nitro, cyano, C1-C10 alkyl, C1-C10 cycloalkyl, C1-C5 fluoroalkyl.

[0015] the substituents of the substituted alkyl are selected from at least one of the group consisting of hydroxyl, carboxyl, amino, C1-C10 alkylamino, mercapto, C1-C10 alkoxy, -COOR0, -CONR0, halogen, nitro, cyano, C1-C10 cycloalkyl, C1-C5 alkyl, C1-C5 fluoroalkyl.

[0016] each of R0 is independently C1-C10 alkyl. Specifically, each of R0 can also be independently C1-C5 alkyl.

[0017] Specifically, the heterocyclic group is selected from the group consisting of furanyl, thienyl, pyrrolyl, thiazolyl, imidazolyl, pyridyl, pyrazinyl, pyrimidinyl, pyridazinyl, etc.

[0018] In some specific embodiments, the compound,

[0019] R is or

[0020] R1 and R2 are each independently a phenyl group or a phenyl group containing at least one substituent, and n is 0, 2, 3, 4 or 5.

[0021] Each of the substituents is independently selected from hydroxyl, carboxyl, amino, C1-C5 alkylamino, mercapto, C1-C5 alkoxy, halogen, nitro, cyano, aldehyde, C1-C5 alkyl, C1-C5 substituted alkyl, and adamantyl.

[0022] The substituent of the substituted alkyl group is selected from at least one of hydroxyl, carboxyl, amino, mercapto, C1-C5 alkoxy, C1-C5 alkyl, halogen, nitro, cyano, C1-C10 cycloalkyl, and C1-C5 fluoroalkyl.

[0023] In some specific embodiments, the compound,

[0024] R is or

[0025] R1 and R2 are each independently a phenyl group or a phenyl group containing at least one substituent, and n is 0, 2, 3, 4 or 5.

[0026] Each of the substituents is independently selected from hydroxyl, methoxy, trifluoromethyl, halogen, C1-C5 alkyl, amino, and adamantyl.

[0027] The present invention provides a pharmaceutical composition comprising the above-described compounds or pharmaceutically acceptable salts thereof.

[0028] The pharmaceutical composition further comprises a carrier and / or pharmaceutical excipients.

[0029] The carrier and pharmaceutical excipients mentioned are commonly used carriers and pharmaceutical excipients in drug preparation.

[0030] The compounds described can be prepared into drugs in the form of tablets, capsules, pills, injections, drops, granules, suppositories, aerosols, sprays, powder inhalers, syrups, tinctures, lotions, films, etc.

[0031] The application of the compound in the preparation of HSP90 inhibitors, HIF-1 inhibitors, drugs for inhibiting tumor development, and anti-angiogenic drugs.

[0032] The HSP90 inhibitor is either a selective HSP90 inhibitor or a non-selective HSP90 inhibitor.

[0033] C1-C10, C1-C5, and C1-C13 respectively represent carbon atoms numbered 1-10, 1-5, and 1-13.

[0034] The beneficial effects of this invention are as follows: This invention utilizes Discovery Studio to reverse-target 7-hydroxyspiroin A, which exhibits good HIF-1 inhibitory activity. HSP90, associated with the HIF-1 pathway, was found to be among the top-scoring targets. Molecular docking of 7-hydroxyspiroin A and HSP90 revealed that the 7-hydroxyl group acts as a hydrogen bond acceptor, playing a crucial hydrogen bond role with HSP90. Given the long and low-yield synthetic route of 7-hydroxyspiroin A, a series of 2-triazolylethane-1-one / alcohol derivatives were designed and synthesized. The cytotoxicity and HIF-1 inhibitory activity of the synthesized compounds were tested, revealing that these compounds retained the low toxicity of 7-hydroxyspiroin A while exhibiting good inhibitory activity against HIF-1. Subsequently, the anti-tumor malignant progression ability of the selected compounds was tested, showing that these compounds can effectively inhibit the invasion and clonal proliferation of HeLa cells. Furthermore, they can effectively inhibit the hypoxic transcriptional activity of VEGF and the invasion of HUVEC cells. In in vivo antitumor experiments, the hydrochloride form of compound 18 was found to effectively inhibit breast cancer in mice, and the inhibitory effect was enhanced when used in combination with cisplatin.

[0035] This type of derivative overcomes the problems of long synthetic routes and low yields of 7-hydroxyspiroin A, while exhibiting low toxicity, effective inhibition of HIF-1, and the ability to inhibit cancer cell development at lower concentrations, greatly improving drug-likeness and overcoming the problem of low drug-likeness of the natural product 7-hydroxyspiroin A. Attached Figure Description

[0036] Figure 1 This is a graph showing the inhibitory activity of compounds 4f and 10a (i.e., compound 18) against cervical cancer cell invasion.

[0037] Figure 2 This is a diagram showing the inhibitory activity of compounds 4f and 10a (i.e., compound 18) on the migration of human umbilical vein endothelial cells.

[0038] Figure 3 This is a diagram showing the inhibitory activity of compounds 4f and 10a (i.e., compound 18) on the clonal proliferation of cervical cancer cells.

[0039] Figure 4 This is a diagram showing the inhibitory activity of compounds 4f and 10a (i.e., compound 18) on VEGF hypoxic transcriptional activity.

[0040] Figure 5 This is a diagram showing the inhibitory activity of compounds 4f and 10a (i.e., compound 18) on VEGF hypoxic transcriptional activity.

[0041] Figure 6This is a schematic diagram of the docking of 10a and Hsp90 molecules.

[0042] Figure 7 This is a schematic diagram of the docking of compound 33 with Hsp90 molecules.

[0043] Figure 8 The tumor growth curves for each group are shown.

[0044] Figure 9 Images of the in situ tumors removed in each group.

[0045] Figure 10 The image shows the weight of the resected tumor in situ for each group.

[0046] Figure 11 This is a graph showing the weight of mice. Detailed Implementation

[0047] In some specific compounds of Formula I, R1 is phenyl, 4-methoxyphenyl, 4-hydroxyphenyl, 3-methoxyphenyl, 3-hydroxyphenyl, 2-methoxyphenyl, 2-hydroxyphenyl, 4-fluorophenyl, 4-methylphenyl, 4-ethylphenyl, 4-tert-butylphenyl, 4-trifluoromethylphenyl, 4-aminophenyl, 2,4-dimethoxyphenyl, 3,5-dimethoxyphenyl, 3,4,5-trimethoxyphenyl, or 4-adamantylphenyl. R2 is 4-hydroxyphenyl or 4-methoxyphenyl.

[0048] In some specific compounds of formula I, R1 is phenyl, 2-methoxyphenyl, 4-aminophenyl, 2,4-dimethoxyphenyl, 3,5-dimethoxyphenyl, or 4-adamantylphenyl. R2 is 4-hydroxyphenyl or 4-methoxyphenyl.

[0049] Some specific compounds of formula I have the following structures:

[0050]

[0051]

[0052] Example 1: Computer-Aided Drug Design

[0053] Using the reverse target search module in Discovery Studio, reverse target search was performed on 7-hydroxyspiroin A. HSP90, which is related to HIF-1, was found to be among the top-scoring targets. Molecular docking of 7-hydroxyspiroin A with HSP90 was performed to identify key interactions and a structural modification scheme for 7-hydroxyspiroin A was proposed.

[0054] Example 22 - Synthesis of triazolyl ethane-1-one / alcohol derivatives

[0055]

[0056] The synthesis of 2-triazolethane-1-one / alcohol derivatives is shown in the above formula.

[0057] 2-Bromo-1-(4-methoxyphenyl)ethyl ketone (200 mg, 0.88 mmol) and sodium azide (171 mg, 2.63 mmol) were added to a round-bottom flask, and Acetone:H₂O was added in a 2:1 ratio. The reaction was carried out at room temperature for 3 h. The mixture was extracted with ethyl acetate and water, and the resulting organic layer was concentrated under reduced pressure and subjected to column chromatography to give the product 2-azido-1-(4-methoxyphenyl)ethyl ketone.

[0058] The brominated compound and PdCl2(PPh3)2 were added to a mixed solvent of water and triethylamine. After thorough mixing, trimethylsilaneacetylene was added, and the mixture was heated to 60°C. After the reaction was complete, the mixture was extracted with ethyl acetate and water. The resulting organic layer was purified by vacuum distillation to remove the solvent, followed by column chromatography to obtain intermediate 1. Intermediate 1 was mixed with methanol, and then K2CO3 was added. The mixture was stirred at room temperature for 3 hours, and then extracted with ethyl acetate and water. The resulting organic layer was purified by vacuum distillation to remove the solvent, giving intermediate 2. Intermediate 2 was reacted with a compound containing an azide group to give the product 2-triazolethane-1-one. The carbonyl group was then reduced to give the product 2-triazolethane-1-ol.

[0059] Synthesis of compound 1 (1-(4-methoxyphenyl)-2-(4-(4-methoxyphenyl)-1H-1,2,3-triazol-1-yl)ethan-1-one)

[0060] 4-Ethynyl anisole (200 mg, 1.52 mmol), 2-azido-1-(4-methoxyphenyl)ethyl ketone (304 mg, 1.59 mmol), and benzoic acid (37 mg, 0.30 mmol) were added to a round-bottom flask, 10 mL of H₂O was added, and the mixture was stirred under N₂ protection. Cu₂O (22 mg, 0.15 mmol) was then added to initiate the reaction. After 2 h, the mixture was extracted with CH₂Cl₂ / H₂O. The resulting organic layer was concentrated under reduced pressure and separated by column chromatography to give 437 mg of a white solid, with a yield of 89%.

[0061] 1H NMR (600MHz, CDCl3) δ8.01 (d, J=8.9Hz, 2H), 7.86 (s, IH), 7.79 (d, J=8.8Hz, 2 H), 7.01 (d, J=8.9Hz, 2H), 6.97 (d, J=δ8.8Hz, 2H), 5.82 (s, 2H), 3.91 (s, 3H), 3.85 (s, 3H).13C-NMR (101MHz, CDCl3) δ188.72, 164.63, 159.62, 130.63, 127 .12, 126.97, 123.30, 114.39, 114.23, 55.65, 55.33, 55.21.HRMS(m / z)[M+H] + calcd forC 18 H 17 N3O3323.1270 found 323.1337.

[0062] Synthesis of compound 2 (1-(4-hydroxyphenyl)-2-(4-(4-hydroxyphenyl)-1H-1,2,3-triazol-1-yl)ethan-1-one)

[0063] Compound 1 (200 mg, 0.62 mmol) was added to a round-bottom flask, followed by 10 mL of CH2Cl2. The mixture was stirred in an ice bath under N2 protection, and boron tribromide (352 μl, 3.72 mmol) was slowly added dropwise. The reaction was monitored by thin-layer chromatography. After 6 h, the reaction was completed. The mixture was extracted with CH2Cl2 / H2O, and the resulting organic layer was concentrated under reduced pressure. The organic layer was then separated by column chromatography to obtain 40 mg of a white solid, with a yield of 22%.

[0064] 1 H-NMR (600MHz, DMSO-d6) δ10.61 (s, 1H), 9.58 (s, 1H), 8.31 (s, 1H), 7.97 (d, J=8.4Hz, 2H ), 7.66 (d, J=8.2Hz, 2H), 6.93 (d, J=8.4Hz, 2H), 6.84 (d, J=8.2Hz, 2H), 6.08 (s, 2H).13C NMR (151MHz, DMSO-d6) δ190.56, 163.43, 157.66, 146.97, 131.33, 126.99, 126.11, 122.29, 122.16, 116.11, 116.02, 60.22.HRMS (m / z) [M+H] + calcd forC 16 H 13 N3O3296.0957 found 296.1022.

[0065] Synthesis of compound 3 (1-(4-methoxyphenyl)-2-(4-(3-methoxyphenyl)-1H-1,2,3-triazo1-1-yl)ethan-1-one)

[0066] 3-Ethynyl anisole (200 mg, 0.52 mmol), 2-azido-1-(4-methoxyphenyl)ethyl ketone (304 mg, 1.59 mmol), and benzoic acid (37 mg, 0.30 mmol) were added to a round-bottom flask, 10 mL of H₂O was added, and the mixture was stirred under N₂ protection. Cu₂O (22 mg, 0.15 mmol) was then added to initiate the reaction. After 2 h, the mixture was extracted with CH₂Cl₂ / H₂O. The resulting organic layer was concentrated under reduced pressure and separated by column chromatography to give 442 mg of a white solid, with a yield of 90%.

[0067] 1H-NMR (600MHz, DMSO-d6) δ8.56 (s, 1H), 8.09 (d, J=8.4Hz, 2H), 7.46 (s, 1H), 7.45 (s, 1H), 7.38 (t, J=7.9Hz , 1H), 7.14 (d, J=8.4Hz, 2H), 6.92 (d, J=7.7Hz, 1H), 6.20 (s, 2H), 3.89 (s, 3H), 3.82 (s, 3H).HRMS (m / z) [M+H] + calcd for C 1s H 17 N3O3324.1270found324.1339.13C-NMR (600MHz, DMSO-d6) δ190.88, 164.41, 160.16, 146.61, 1 32.58, 131.12, 130.57, 127.40, 123.84, 117.93, 114.72, 114.08, 110.72, 55.18, 56.11, 55.58.

[0068] Synthesis of compound 4 (1-(4-hydroxyphenyl)-2-(4-(3-hydroxyphenyl)-1H-1,2,3-triazol-1-yl)ethan-1-one)

[0069] Compound 3 (200 mg, 0.62 mmol) was added to a round-bottom flask, followed by 10 mL of CH2Cl2. The mixture was stirred in an ice bath under N2 protection, and boron tribromide (352 μl, 3.72 mmol) was slowly added dropwise. The reaction was monitored by thin-layer chromatography. After 6 h, the reaction was completed. The mixture was extracted with CH2Cl2 / H2O, and the resulting organic layer was concentrated under reduced pressure. The organic layer was then separated by column chromatography to obtain 40 mg of a white solid, with a yield of 22%.

[0070] 1 H-NMR (600MHz, DMSO-d6) δ10.62 (s, 1H), 9.57 (s, 1H), 8.43 (s, 1H), 7.98 (d, J=8.6Hz, 2 H), 7.30 (s, 1H), 7.28-7.20 (m, 2H), 6.94 (d, J = 8.6Hz, 2H), 6.74 (dt, J = 7.1, 2.2Hz, 1H), 6.10 (s, 2H).13C-NMR (151MHz, DMSO-d6) δ190.52, 163.49, 158.26, 146.77, 132.48, 131 .37, 130.49, 126.08, 123.52, 116.49, 116.05, 115.34, 112.28, 55.91.HRMS(m / z)[M+H] + calcd for C 16 H 13 N3O3296.0957 found 296.1040.

[0071] Synthesis of compound 5 (1-(4-methoxyphenyl)-2-(4-(2-methoxyphenyl)-1H-1,2,3-triazol-1-yl)ethan-1-one)

[0072] 2-Ethynyl anisole (200 mg, 1.52 mmol), 2-azido-1-(4-methoxyphenyl)ethyl ketone (304 mg, 1.59 mmol), and benzoic acid (37 mg, 0.30 mmol) were added to a round-bottom flask, 10 mL of H₂O was added, and the mixture was stirred under N₂ protection. Cu₂O (22 mg, 0.15 mmol) was then added to initiate the reaction. After 2 h, the mixture was extracted with CH₂Cl₂ / H₂O. The resulting organic layer was concentrated under reduced pressure and separated by column chromatography to give 138 mg of a white solid, with a yield of 28%.

[0073] 1H-NMR (400MHz, DMSO-d6) δ8.41 (s, 1H), 8.20 (dd, J=7.4, 1.6Hz, 1H), 8.08 (d, J=8.9Hz, 2H), 7.427.2 7 (m, 1H), 7.15 (d, J=11.2Hz, 1H), 7.14 (d, J=8.9Hz, 2H), 7.08 (td, J=7.5, 1.1Hz, 1H), 6.17 (s, 2H), 3. 91 (s, 3H), 3.89 (s, 3H). 13C-NMR (151MHz, DMSO-d6) δ191.06, 164.36, 155.77, 142.10, 131.07, 129. 32, 127.51, 126.93, 126.21, 121.15, 119.60, 114.71, 112.02, 56.16, 55.92.HRMS(m / z)[M+H]+calcd for C18H17N3O3324.1270found324.1348.

[0074] Synthesis of compound 6 (2-(4-(4-fluorophenyl)-1H-1,2,3-triazol-1-yl)-1-(4-methoxyphenyl)ethan-1-one)

[0075] 4-Fluorophenylacetylene (200 mg, 1.66 mmol), 2-azido-1-(4-methoxyphenyl)ethyl ketone (333 mg, 1.74 mmol), and benzoic acid (40 mg, 0.33 mmol) were added to a round-bottom flask, 10 mL of H₂O was added, and the mixture was stirred under N₂ protection. Cu₂O (24 mg, 0.17 mmol) was then added to initiate the reaction. After 2 h, the mixture was extracted with CH₂Cl₂ / H₂O. The resulting organic layer was concentrated under reduced pressure and separated by column chromatography to give 330 mg of a white solid, with a yield of 64%.

[0076] 1H NMR (400MHz, Chloroform-d) δ8.00 (d, J=8.5Hz, 2H), 7.93 (s, 1H), 7.83 (dd, J=8.5, 5.3Hz, 2H), 7.12 (t, J=8.3Hz, 2H), 7.00 (d, J=8.4Hz, 2H), 5.84 (s, 2H), 3.90 (s, 3H). 13C-NMR (151MHz, Chloroform-d) δ188.65, 164.73, 163.53, 161.90, 130.66, 127 .59, 127.54, 126.90, 115.91, 115.76, 114.45, 55.68, 55.24.HRMS(m / z)[M+H] + calcd forC 17 H 14 FN3O2312.1070found 312.1136.

[0077] Synthesis of compound 7 (2-(4-(4-fluorophenyl)-1H-1,2,3-triazol-1-yl)-1-(4-hydroxyphenyl)ethan-1-one)

[0078] Compound 6 (200 mg, 0.64 mmol) was added to a round-bottom flask, 6 mL of HBr was added, N2 protection was applied, and the mixture was refluxed at 120 °C. Thin-layer chromatography was used for monitoring. After 6 h, the reaction was stopped, quenched with water, and extracted with CH2Cl2 / saturated NaHCO3. The resulting organic layer was concentrated under reduced pressure and separated by column chromatography to obtain 65 mg of white solid, with a yield of 36%.

[0079] 1H NMR (600MHz, DMSO-d6) δ10.45 (s, 1H), 8.50 (s, 1H), 7.98 (d, J=8.5Hz, 2H), 7.91 (dd , J=8.5, 5.5Hz, 2H), 7.30 (t, J=8.8Hz, 2H), 6.93 (d, J=8.4Hz, 2H), 6.12 (s, 2H).13C NMR (151MHz, DMSO-d6) δ190.49, 163.53, 145.85, 131.38, 127.66, 127.60, 126.06, 123.50, 116.43, 116.28, 116.06, 55.96.HRMS (m / z) [M+H] + calcd forC 16 H 12 FN3O2298.0914 found 298.0981.

[0080] Synthesis of compound 8 (1-(4-methoxyphenyl)-2-(4-phenyl-1H-1,2,3-triazol-1-yl)ethan-1-one)

[0081] Phenylacetylene (215 μl, 1.96 mmol), 2-azido-1-(4-methoxyphenyl)ethyl ketone (393 mg, 2.06 mmol), and benzoic acid (47 mg, 0.392 mmol) were added to a round-bottom flask. 10 mL of H₂O was added, and the mixture was stirred under N₂ protection. Cu₂O (27 mg, 0.20 mmol) was then added to initiate the reaction. After 2 h, the mixture was extracted with CH₂Cl₂ / H₂O. The resulting organic layer was concentrated under reduced pressure and separated by column chromatography to give 241 mg of a white solid, with a yield of 42%.

[0082] 1 H-NMR (400MHz, CDCl3) δ7.99 (d, J=8.9Hz, 2H), 7.95 (s, 1H), 7.86 (d, J=6.9Hz, 1H ), 7.43 (t, J=7.5Hz, 2H), 7.37-7.30 (m, 1H), 7.00 (d, J=8.9Hz, 2H), 5.83 (s, 2H), 3 .90 (s, 3H).13C-NMR (151MHz, CDCl3) δ188.67, 164.61, 148.10, 130.60, 130.54, 128.81, 128.14, 126.88, 125.79, 121.52, 114.37, 55.64, 55.16.HRMS(m / z)[M+H] + calcd forC 17 H 15 N3O2294.1164 found 294.1230.

[0083] Synthesis of compound 9 (1-(4-hydroxyphenyl)-2-(4-phenyl-1H-1,2,3-triazol-1-yl)ethan-1-one)

[0084] Compound 8 (200 mg, 0.68 mmol) was added to a round-bottom flask, 6 mL of HBr was added, N2 protection was applied, and the mixture was refluxed at 120 °C. Thin-layer chromatography was used for monitoring. After 6 h, the reaction was stopped, quenched with water, and extracted with CH2Cl2 / saturated NaHCO3. The resulting organic layer was concentrated under reduced pressure and separated by column chromatography to obtain 110 mg of white solid, yield 58%.

[0085] 1H-NMR (600MHz, DMSO-d6) δ10.58 (s, 2H), 8.50 (s, 1H), 7.98 (d, J=8.2Hz, 2H), 7. 87 (d, J=7.5Hz, 2H), 7.46 (t, J=7.6Hz, 2H), 7.35 (s, 1H), 6.94 (d, J=8.8Hz, 2H), 6.12 (s, 2H).13C-NMR (151MHz, DMSO-d6) δ189.88, 162.90, 146.12, 130.77, 130 .72, 128.82, 127.70, 125.54, 125.03, 122.97, 115.48, 55.34.HRMS(m / z)[M+H] + calcd forC 16 H 13 N3O2280.1008 found 280.1076.

[0086] Synthesis of compound 10 (1-(4-methoxyphenyl)-2-(4-(p-tolyl)-1H-1,2,3-triazol-1-yl)ethan-1-one)

[0087] 4-Tolylacetylene (200 mg, 1.72 mmol), 2-azido-1-(4-methoxyphenyl)ethyl ketone (304 mg, 1.81 mmol), and benzoic acid (42 mg, 0.34 mmol) were added to a round-bottom flask, 10 mL of H₂O was added, and the mixture was stirred under N₂ protection. Cu₂O (25 mg, 0.17 mmol) was then added to initiate the reaction. After 2 h, the mixture was extracted with CH₂Cl₂ / H₂O. The resulting organic layer was concentrated under reduced pressure and separated by column chromatography to give 125 mg of a white solid, with a yield of 24%.

[0088] 1 H NMR (600MHz, Acetone-d6) δ8.33 (s, 1H), 8.13 (d, J = 8.9Hz, 2H), 7.82 (d, J = 7.7Hz, 2 H), 7.26 (d, J=7.7Hz, 2H), 7.13 (d, J=8.9Hz, 2H), 6.12 (s, 2H), 3.94 (s, 3H), 2.36 (s, 3H).13C-NMR (151MHz, Acetone-d6) δ190.51, 165.31, 147.94, 138.18, 131.38, 130. 24, 129.61, 128.36, 126.18, 122.80, 115.02, 56.21, 56.10, 21.23.HRMS(m / z)[M+H] + calcd forC18 H 17 N3O2308.1321 found 308.1402.

[0089] Synthesis of Compound 11 (1-(4-hydroxyphenyl)-2-(4-(p-tolyl)-1H-1,2,3-triazol-1-yl)ethan-1-one): Compound 10 (200 mg, 0.65 mmol) was added to a round-bottom flask, 6 mL of HBr was added, N2 protection was applied, and the mixture was refluxed at 120 °C. Thin-layer chromatography was used for monitoring. After 6 h, the reaction was stopped, quenched with water, and extracted with CH2Cl2 / saturated NaHCO3. The resulting organic layer was concentrated under reduced pressure and separated by column chromatography to give 50 mg of white solid, yield 26%.

[0090] 1 H-NMR (600MHz, DMSO-d6) δ10.60 (s, 1H), 8.44 (s, 1H), 7.98 (d, J=8.4Hz, 2H), 7.76 ( d, J=7.7Hz, 2H), 7.27 (d, J=7.7Hz, 2H), 6.93 (d, J=8.4Hz, 2H), 6.10 (s, 2H), 2.34 (s , 3H).13C-NMR (151MHz, DMSO-d6) δ190.48, 163.53, 146.74, 137.56, 131.34, 129.9 5, 128.52, 126.06, 125.54, 123.13, 116.06, 55.88, 21.31.HRMS(m / z)[M+H]+calcd for C17H15N3O2294.1164found 294.1234.

[0091] Synthesis of compound 12 (2-(4-(4-ethylphenyl)-1H-1,2,3-triazol-1-yl)-1-(4-methoxyphenyl)ethan-1-one)

[0092] 4-Ethylphenylacetylene (200 mg, 1.72 mmol), 2-azido-1-(4-methoxyphenyl)ethyl ketone (304 mg, 1.81 mmol), and benzoic acid (42 mg, 0.34 mmol) were added to a round-bottom flask, 10 mL of H₂O was added, and the mixture was stirred under N₂ protection. Cu₂O (25 mg, 0.17 mmol) was then added to initiate the reaction. After 2 h, the mixture was extracted with CH₂Cl₂ / H₂O. The resulting organic layer was concentrated under reduced pressure and separated by column chromatography to give 125 mg of a white solid, with a yield of 24%.

[0093] 1H-NMR (600MHz, Acetone-d6) δ8.32 (s, 1H), 8.12 (d, J=8.9Hz, 2H), 7.84 (d, J=7.9Hz, 2H), 7.29 (d, J=7.9H z, 2H), 7.12 (d, J=8.9Hz, 2H), 6.12 (s, 2H), 3.93 (s, 3H), 2.67 (q, J=7.7Hz, 2H), 1.24 (t, J=7.7Hz, 2H).13C NMR (151MHz, Chloroform-d) δ188.64, 164.69, 147.97, 144.63, 130.68, 128.38, 127 .53, 126.92, 125.92, 121.42, 114.42, 55.67, 55.38, 28.70, 15.50.HRMS(m / z)[M+H] + calcd for C 19 H 19 N3O2 322.1477 found 322.1559.

[0094] Synthesis of compound 13 (2-(4-(4-ethylphenyl)-1H-1,2,3-triazol-1-y1)-1-(4-hydroxyphenyl)ethan-1-one)

[0095] Compound 12 (200 mg, 0.65 mmol) was added to a round-bottom flask, 6 mL of HBr was added, N2 protection was applied, and the mixture was refluxed at 120 °C. Thin-layer chromatography was used for monitoring. After 6 h, the reaction was stopped, quenched with water, and extracted with CH2Cl2 / saturated NaHCO3. The resulting organic layer was concentrated under reduced pressure and separated by column chromatography to obtain 42 mg of white solid, yield 21%.

[0096] 1 H-NMR (600MHz, DMSO-d6) δ10.58 (s, 1H), 8.44 (s, 1H), 7.98 (d, J=8.8Hz, 2H), 7.78 (d, J=7.8Hz, 2H), 7.86 (d, J=6. 9Hz, 1H), 7.30 (d, J=7.8Hz, 2H), 6.93 (d, J=8.8Hz, 2H), 6.10 (s, 2H), 2.64 (q, J=7.6Hz, 2H), 1.21 (t, J=7.6Hz, 3H). 13C NMR (151MHz, DMSO-d6) δ189.90, 162.89, 146.18, 143.32, 130.76, 128.21, 12 8.17, 125.55, 125.06, 122.57, 115.47, 55.30, 27.83, 15.37.HRMS(m / z)[M+H] + calcd for C 17 H 15 N3O2308.1321found308.1387.

[0097] Synthesis of compound 14 (2-(4-(4-(tert-butyl)phenyl)-1H-1,2,3-triazol-1-yl)-1-(4-methoxyphenyl)ethan-1-one)

[0098] 4-tert-butylphenylacetylene (228 μl, 1.26 mmol), 2-azido-1-(4-methoxyphenyl)ethyl ketone (252 mg, 1.32 mmol), and benzoic acid (31 mg, 0.25 mmol) were added to a round-bottom flask, 10 mL of H₂O was added, and the mixture was stirred under N₂ protection. Cu₂O (18 mg, 0.13 mmol) was then added to initiate the reaction. After 2 h, the mixture was extracted with CH₂Cl₂ / H₂O. The resulting organic layer was concentrated under reduced pressure and separated by column chromatography to give 374 mg of a white solid, with a yield of 85%.

[0099] 1 H-NMR (400MHz, CDCl3) δ8.01 (d, J=8.8Hz, 2H), 7.80 (d, J=8.0Hz, 2H), 7.46 (d, J=7.9Hz, 2H), 7.26 (s, 1H), 7.00 (d, J=8.8Hz, 2H), 5.83 (s, 2H), 3.91 (s, 3H), 1. 35(s, 9H).13C-NMR (151MHz, CDCl3) δ188.72, 164.66, 151.28, 130.65, 127.78, 127.05, 125.74, 125.58, 114.42, 55.65, 55.21, 34.69, 31.32.HRMS(m / z)[M+H] + calcd forC 21 H 23 N3O2 350.1790 found 350.1851.

[0100] Synthesis of compound 15 (2-(4-(4-(tert-butyl)phenyl)-1H-1,2,3-triazol-1-yl)-1-(4-methoxyphenyl)ethan-1-one)

[0101] Compound 14 (200 mg, 0.57 mmol) was added to a round-bottom flask, 6 mL of HBr was added, N2 protection was applied, and the mixture was refluxed at 120 °C. The reaction was monitored by thin-layer chromatography. After 6 h, the reaction was quenched with water and extracted with CH2Cl2 / saturated NaHCO3. The resulting organic layer was concentrated under reduced pressure and separated by column chromatography to obtain 171 mg of white solid, with a yield of 89%.

[0102] 1 H-NMR (600MHz, DMSO-d6) δ10.59 (s, 1H), 8.45 (s, 1H), 7.98 (d, J=8.7Hz, 2H), 7.78 (d, J= 8.4Hz, 2H), 7.48 (d, J=8.4Hz, 1H), 6.93 (d, J=8.7Hz, 2H), 6.10 (s, 2H), 1.31 (s, 9H).13C NMR (151MHz, DMSO-d6) δ190.53, 163.49, 150.79, 146.69, 131.36, 128.56, 12 6.14, 125.43, 123.24, 116.06, 55.88, 49.07, 34.84, 31.56.HRMS(m / z)[M+H] + calcd forC 20 H 21 N3O2336.1634 found 308.1387.

[0103] Synthesis of compound 16 (1-(4-methoxyphenyl)-2-(4-(4-(trifluoromethyl)phenyl)-1H-1,2,3-triazol-1-yl)ethan-1-one)

[0104] 4-Trifluoromethylphenylacetylene (292 μl, 1.18 mmol), 2-azido-1-(4-methoxyphenyl)ethyl ketone (237 mg, 1.24 mmol), and benzoic acid (29 mg, 0.24 mmol) were added to a round-bottom flask, 10 mL of H₂O was added, and the mixture was stirred under N₂ protection. Cu₂O (17 mg, 0.12 mmol) was then added to initiate the reaction. After 2 h, the mixture was extracted with CH₂Cl₂ / H₂O. The resulting organic layer was concentrated under reduced pressure and separated by column chromatography to give 345 mg of a white solid, with a yield of 81%.

[0105] 1 H-NMR (600MHz, DMSO-d6) δ8.70 (s, 1H), 8.11 (d, J=8.0Hz, 2H), 8.09 (d, J=8.9 Hz, 2H), 7.83 (d, J=8.0Hz, 1H), 7.15 (d, J=8.9Hz, 2H), 6.23 (s, 2H), 3.89 (s, 3H ).13C-NMR (151MHz, DMSO-d6) δ190.20, 163.96, 144.85, 134.70, 130.62, 126 .89, 125.90, 125.88, 125.64, 124.33, 114.23, 55.70, 55.68.HRMS(m / z)[M+H] + calcd forC 18 H 14 F3N3O2362.1038 found 362.1123.

[0106] Synthesis of compound 17 (1-(4-hydroxyphenyl)-2-(4-(4-(trifluoromethyl)phenyl)-1H-1,2,3-triazol-1-yl)ethan-1-one)

[0107] Compound 16 (200 mg, 0.55 mmol) was added to a round-bottom flask, 6 mL of HBr was added, N2 protection was applied, and the mixture was refluxed at 120 °C. Thin-layer chromatography was used for monitoring. After 6 h, the reaction was stopped, quenched with water, and extracted with CH2Cl2 / saturated NaHCO3. The resulting organic layer was concentrated under reduced pressure and separated by column chromatography to obtain 91 mg of white solid, with a yield of 48%.

[0108] 1 H-NMR (600MHz, DMSO-d6) δ10.60 (s, 1H), 8.68 (s, 1H), 8.10 (d, J=8.0Hz, 2H) , 7.98 (d, J = 8.4Hz, 2H), 7.83 (d, J = 8.0Hz, 1H), 6.94 (d, J = 8.4Hz, 2H), 6.16 ( s, 2H).13C-NMR (151MHz, DMSO-d6) δ190.37, 163.55, 145.35, 135.26, 131.3 9, 126.43, 126.41, 126.16, 126.06, 124.87, 116.08, 56.05.HRMS(m / z)[M+H] + calcd for C 17 H 12 F3N3O2348.0882found348.0952.

[0109] Synthesis of compound 18 (2-(4-(4-aminophenyl)-1H-1,2,3-triazol-1-yl)-1-(4-methoxyphenyl)ethan-1-one)

[0110] 4-Aminophenylacetylene (191 μl, 1.71 mmol), 2-azido-1-(4-methoxyphenyl)ethyl ketone (342 mg, 1.79 mmol), and benzoic acid (47 mg, 0.34 mmol) were added to a round-bottom flask, 10 mL of H₂O was added, and the mixture was stirred under N₂ protection. Cu₂O (24 mg, 0.17 mmol) was then added to initiate the reaction. After 2 h, the mixture was extracted with CH₂Cl₂ / H₂O. The resulting organic layer was concentrated under reduced pressure and separated by column chromatography to give 305 mg of a white solid, with a yield of 58%.

[0111] 1 H-NMR (600MHz, DMSO-d6) δ8.20 (s, 1H), 8.07 (d, J = 8.8Hz, 2H), 7.51 (d, J = 8.5Hz, 2H), 7.13 (d, J=8.8Hz, 1H), 6.62 (d, J=8.5Hz, 2H), 6.10 (s, 2H), 5.23 (s, 2H), 3.88 (s, 3H). 13 C-NMR (151MHz, DMSO-d6) δ191.03, 164.40, 149.07, 147.70, 131.09, 127.56, 126.64, 121.30, 118.95, 114.73, 114.47, 56.18, 55.92.HRMS (m / z) [M+H] + calcd forC 17 H 16 N4O2309.1273 found 309.1358.

[0112] Synthesis of compound 19 (2-(4-(4-aminophenyl)-1H-1,2,3-triazol-1-yl)-1-(4-hydroxyphenyl)ethan-1-one)

[0113] 1,4-Dimethylphenylacetylene (200 mg, 1.23 mmol), 2-azido-1-(4-methoxyphenyl)ethyl ketone (247 mg, 1.30 mmol), and benzoic acid (30 mg, 0.25 mmol) were added to a round-bottom flask, 10 mL of H₂O was added, and the mixture was stirred under N₂ protection. Cu₂O (18 mg, 0.13 mmol) was then added to initiate the reaction. After 2 h, the mixture was extracted with CH₂Cl₂ / H₂O. The resulting organic layer was concentrated under reduced pressure and separated by column chromatography to give 305 mg of a white solid, with a yield of 29%.

[0114] 1 H-NMR (600MHz, CDCl3) δ8.27 (d, J=8.6Hz, 1H), 8.10 (s, 1H), 8.01 (d, J=8.9Hz, 2H), 7.00 (d, J=8.9H z, 2H), 6.63 (dd, J=8.6, 2.4Hz, 2H), 6.54 (d, J=2.4Hz, 1H), 5.81 (s, 2H), 3.90 (s, 6H), 3.86 (s, 3H). 13 C-NMR (151MHz, Chloroform-d) δ189.05, 164.56, 160.66, 156.94, 143.71, 130.67, 128.48, 1 27.17, 123.64, 114.36, 112.65, 104.93, 98.59, 55.62, 55.45, 55.40, 55.15.HRMS(m / z)[M+H] + calcd for C 19 H 19 N3O4354.1376 found 354.1438.

[0115] Synthesis of compound 20 (2-(4-(4-aminophenyl)-1H-1,2,3-triazol-1-yl)-1-(4-hydroxyphenyl)ethan-1-one)

[0116] 3,5-Dimethoxyphenylacetylene (191 μl, 1.71 mmol), 2-azido-1-(4-methoxyphenyl)ethyl ketone (342 mg, 1.79 mmol), and benzoic acid (47 mg, 0.34 mmol) were added to a round-bottom flask, 10 mL of H₂O was added, and the mixture was stirred under N₂ protection. Cu₂O (24 mg, 0.17 mmol) was then added to initiate the reaction. After 2 h, the mixture was extracted with CH₂Cl₂ / H₂O. The resulting organic layer was concentrated under reduced pressure and separated by column chromatography to give 305 mg of a white solid, in 58% yield.

[0117] 1 H-NMR (600MHz, CDCl3) δ8.01 (d, J=8.7Hz, 2H), 7.93 (s, 1H), 7.04 (d, J=2.3Hz, 2H), 7 .01 (d, J=8.7Hz, 2H), 6.46 (t, J=2.4Hz, 1H), 5.83 (s, 2H), 3.91 (s, 3H), 3.85 (s, 6H). 13 C-NMR (151MHz, CDCl3) δ188.64, 164.72, 161.19, 148.09, 132.41, 130.66, 12 6.96, 121.76, 114.44, 103.77, 100.80, 55.65, 55.51, 55.16.HRMS(m / z)[M+H] + calcd forC 19 H 19 N3O4354.1376 found 354.1450.

[0118] Synthesis of compound 212-(4-(3,5-dihydroxyphenyl)-1H-1,2,3-triazol-1-yl)-1-(4-hydroxyphenyl)ethan-1-one)

[0119] Compound 20 (200 mg, 0.57 mmol) was added to a round-bottom flask, 6 mL of HBr was added, N2 protection was applied, and the mixture was refluxed at 120 °C. The reaction was monitored by thin-layer chromatography. After 6 h, the reaction was quenched with water and extracted with CH2Cl2 / saturated Na2HCO3. The resulting organic layer was concentrated under reduced pressure and separated by column chromatography to obtain 81 mg of white solid, with a yield of 46%.

[0120] 1 H NMR (600MHz, DMSO-d6) δ10.59 (s, 1H), 9.35 (s, 2H), 8.33 (s, 1H), 7.97 (d, J=8 .8Hz, 2H), 6.93 (d, J=8.8Hz, 2H), 6.71 (s, 2H), 6.19 (s, 1H), 6.07 (s, 2H).13C NMR (151MHz, DMSO-d6) δ190.51, 163.49, 159.31, 146.97, 132.82, 131.35, 126.07, 123.38, 116.06, 103.83, 102.61, 55.88.HRMS (m / z) [MH] - calcd for C 16 H 13N3O4310.0906found310.2100.

[0121] Synthesis of compound 22 (1-(4-methoxyphenyl)-2-(4-(3,4,5-trimethoxyphenyl)-1H-1,2,3-triazol-1-yl)ethan-1-one)

[0122] 3,4,5-Trimethoxyphenylacetylene (200 mg, 1.04 mmol), 2-azido-1-(4-methoxyphenyl)ethyl ketone (208 mg, 1.09 mmol), and benzoic acid (47 mg, 0.34 mmol) were added to a round-bottom flask, 10 mL of H₂O was added, and the mixture was stirred under N₂ protection. Cu₂O (24 mg, 0.17 mmol) was then added to initiate the reaction. After 2 h, the mixture was extracted with CH₂Cl₂ / H₂O. The resulting organic layer was concentrated under reduced pressure and separated by column chromatography to give 208 mg of a white solid, with a yield of 65%.

[0123] 1 H-NMR (600MHz, CDCl3) δ 8.02 (d, J=8.9Hz, 2H), 7.93 (s, 1H), 7.10 (s, 2H), 7.02 (d, J=8.9Hz, 2H), 5.84 (s, 2H), 3.94 (s, 6H), 3.92 (s, 3H), 3.89 (s, 3H). 13 C-NMR (151MHz, Chloroform-d) δ188.71, 164.77, 153.68, 148.14, 130.67, 126.9 4, 126.21, 121.26, 114.47, 103.09, 60.98, 56.28, 55.67, 55.16.HRMS(m / z)[M+H] + calcd for C 20 H 21 N3O5384.1481found384.1556.

[0124] Synthesis of compound 23 (4-(4-methoxyphenyl)-1-phenyl-1H-1,2,3-triazole)

[0125] Benzeneboronic acid (200 mg, 1.67 mmol), sodium azide, and anhydrous copper sulfate were added to a round-bottom flask under nitrogen protection to generate benzene azide, which reacted with 1-ethynyl-4-methoxybenzene by a click reaction. The mixture was extracted with CH2Cl2 / H2O, and the resulting organic layer was concentrated under reduced pressure and separated by column chromatography to give 360 ​​mg of white solid, with a yield of 86%.

[0126] 1 H NMR (600MHz, Chloroform-d) δ8.11 (s, 1H), 7.84 (d, J=8.7Hz, 2H), 7.81-7.74 (m, 2H ), 7.55 (t, J=7.8Hz, 2H), 7.45 (t, J=7.4Hz, 1H), 7.00 (d, J=8.8Hz, 2H), 3.86 (s, 3H). 13 C NMR (151MHz, DMSO-d6) δ159.77, 147.75, 137.17, 130.42, 129.12, 127.20, 123.25, 120.41, 119.09, 114.91, 55.68.[M+H] + calcd for C 15 H 13 N3O251.1059 found 251.1130.

[0127] Synthesis of compound 24 (4-(1-phenyl-1H-1,2,3-triazol-4-yl)phenol)

[0128] Compound 23 (200 mg, 0.80 mmol) was added to a round-bottom flask, 6 mL of HBr was added, N2 protection was applied, and the mixture was refluxed at 120 °C. Thin-layer chromatography was used for monitoring. After 6 h, the reaction was stopped, quenched with water, and extracted with CH2Cl2 / saturated NaHCO3. The resulting organic layer was concentrated under reduced pressure and separated by column chromatography to obtain 53 mg of white solid, with a yield of 46%.

[0129] 1 H NMR (600MHz, DMSO-d6) δ9.73 (s, 1H), 9.13 (s, 1H), 7.94 (d, J=7.9Hz, 2H), 7.76 (d, J=8.4Hz, 2H), 7.63 (t, J=8.0Hz, 2H), 7.51 (t, J=7.4Hz, 1H), 6.89 (d, J=8.2Hz, 2H). 13 C NMR (151MHz, DMSO-d6) δ158.10, 148.15, 137.23, 130.39, 129.02, 127.28, 121.68, 120.35, 118.61, 116.20.[M+H] + calcd for C 14 H 11 N3O2 37.0902 found 237.0970.

[0130] Synthesis of compound 25 (1-benzyl-4-(4-methoxyphenyl)-1H-1,2,3-triazole)

[0131] The compound bromotoluene (200 mg, 1.18 mmol) was reacted with sodium azide in a round-bottom flask to generate an azide. This azide was then reacted with 1-ethynyl-4-methoxybenzene, extracted with CH₂Cl₂ / saturated NaHCO₃, and the resulting organic layer was concentrated under reduced pressure and separated by column chromatography to give 231 mg of a white solid, with a yield of 74%.

[0132] 1 H NMR (600MHz, Chloroform-d) δ7.73 (d, J=8.2Hz, 2H), 7.59 (s, 1H), 7.41-7.34 ( m, 3H), 7.31 (d, J=6.8Hz, 2H), 6.93 (d, J=8.1Hz, 2H), 5.56 (s, 2H), 3.83 (s, 3H).

[0133] Synthesis of compound 26 (4-(1-benzyl-1H-1,2,3-triazol-4-yl)phenol)

[0134] Compound 25 (200 mg, 0.75 mmol) was added to a round-bottom flask, 6 mL of HBr was added, and the mixture was protected with N2. The mixture was refluxed at 120 °C and monitored by thin-layer chromatography. After 6 h, the reaction was quenched with water and extracted with CH2Cl2 / saturated NaHCo3. The resulting organic layer was concentrated under reduced pressure and separated by column chromatography to give 104 mg of white solid, yield 55%.

[0135] 1 H NMR (600MHz, DMSO-d6) δ9.59 (s, 1H), 8.45 (s, 1H), 7.65 (dd, J=8.7, 2.4Hz, 2H), 7.41 -7.37 (m, 2H), 7.34 (dt, J=9.4, 2.9Hz, 3H), 6.82 (dd, J=8.7, 2.7Hz, 2H), 5.61 (s, 2H). 13 C NMR (101MHz, DMSO-d6) δ157.75, 147.46, 136.63, 129.26, 128.59, 128.33, 127.06, 122.17, 120.63, 116.08, 53.40.[M+H] + calcd for C 15 H 13 N3O 251.1059 found 251.1127.

[0136] Synthesis of compound 27 (4-(4-methoxyphenyl)-1-phenethyl-1H-1,2,3-triazole)

[0137] The compound bromoethylbenzene (200 mg, 1.08 mmol) was reacted with sodium azide in a round-bottom flask to generate an azide. It was then reacted with 1-ethynyl-4-methoxybenzene, extracted with CH₂Cl₂ / saturated NaHCO₃, and the resulting organic layer was concentrated under reduced pressure and separated by column chromatography to give 286 mg of a white solid, with a yield of 74%.

[0138] 1 H NMR (600MHz, Chloroform-d) δ7.69 (d, J=8.7Hz, 2H), 7.39 (s, 1H), 7.30 (dd, J=8.1, 6.6Hz, 2H), 7.27-7.24 (m, 1H ), 7.13 (d, J=6.8Hz, 2H), 6.93 (d, J=8.7Hz, 2H), 4.61 (dd, J=8.1, 6.5Hz, 2H), 3.83 (s, 3H), 3.24 (t, J=7.3Hz, 2H). 13 C NMR (151MHz, DMSO-d6) δ159.43, 146.53, 138.15, 129.18, 128.92, 127.07, 126.89, 123.91, 120.85, 114.79, 55.62, 51.02, 36.04.[M+H] + calcd for C 17 H 17 N3O 279.1372 found 279.1443.

[0139] Synthesis of compound 28 (4-(1-phenethyl-1H-1,2,3-triazol-4-yl)phenol)

[0140] Compound 27 (200 mg, 0.71 mmol) was added to a round-bottom flask, 6 mL of HBr was added, N2 protection was applied, and the mixture was refluxed at 120 °C. Thin-layer chromatography was used for monitoring. After 6 h, the reaction was stopped, quenched with water, and extracted with CH2Cl2 / saturated NaHCO3. The resulting organic layer was concentrated under reduced pressure and separated by column chromatography to obtain 179 mg of white solid, with a yield of 95%.

[0141] 1H NMR (600MHz, DMSO-d6) δ9.58 (s, 1H), 8.33 (s, 1H), 7.60 (d, J = 7.4Hz, 2H), 7.28 (t, J = 7.5Hz, 2H ), 7.22 (d, J = 7.8Hz, 3H), 6.81 (d, J = 7.6Hz, 2H), 4.62 (t, J = 7.4Hz, 2H), 3.20 (t, J = 7.4Hz, 2H). 13 C NMR (151MHz, DMSO-d6) δ157.69, 146.92, 138.16, 129.18, 128.91, 127.06, 126.96, 122.33, 120.39, 116.09, 50.97, 36.06.[M+H] + calcd for C 16 H 15 N3O265.1225 found 265.1291.

[0142] Synthesis of compound 29 (2-(4-(4-methoxyphenyl)-1H-1,2,3-triazol-1-yl)-1-phenylethan-1-one)

[0143] The compound bromoacetophenone (200 mg, 1 mmol) reacted with sodium azide in a mixture of water and acetone to form an azide, which then underwent a click reaction with 1-ethynyl-4-methoxybenzene. The mixture was extracted with CH2Cl2 / saturated NaHCO3, and the resulting organic layer was concentrated under reduced pressure and separated by column chromatography to give 67 mg of white solid, with a yield of 23%.

[0144] 1 H NMR (600MHz, DMSO-d6) δ8.42 (s, 1H), 8.11 (d, J = 7.7Hz, 2H), 7.81 (d, J = 8.3Hz, 2H), 7.76 ( t, J=7.4Hz, 1H), 7.63 (t, J=7.6Hz, 2H), 7.04 (d, J=8.3Hz, 2H), 6.25 (s, 2H), 3.80 (s, 3H). 13 C NMR (151MHz, DMSO-d6) δ192.77, 159.49, 146.73, 134.77, 134.62, 129.50, 128.71, 126.99, 123.84, 122.58, 114.84, 56.42, 55.62.[M+H] + calcd forC 17 H 15 N3O2293.1164 found 293.1238.

[0145] Synthesis of compound 30 (2-(4-(4-hydroxyphenyl)-1H-1,2,3-triazol-1-yl)-1-phenylethan-1-one)

[0146] Compound 29 (200 mg, 0.68 mmol) was added to a round-bottom flask, 6 mL of HBr was added, and the mixture was protected with N2. The mixture was refluxed at 120 °C and monitored by thin-layer chromatography. After 6 h, the reaction was quenched with water and extracted with CH2Cl2 / saturated NaHCO3. The resulting organic layer was concentrated under reduced pressure and separated by column chromatography to obtain 176 mg of white solid, with a yield of 93%.

[0147] 1 H NMR (600MHz, DMSo-d6) δ9.61 (s, 1H), 8.33 (s, 1H), 8.10 (d, J = 7.7Hz, 2H), 7.75 (t, J = 7.5Hz, 1H) , 7.68 (dd, J=8.7, 2.7Hz, 2H), 7.62 (t, J=7.7Hz, 2H), 6.85 (dd, J=8.7, 2.8Hz, 2H), 6.22 (s, 2H). 13 C NMR (151MHz, DMSo-d6) δ192.79, 157.74, 147.09, 134.76, 134.62, 129.49, 128.70, 127.05, 122.25, 122.13, 116.15, 56.39.[M+H] + calcd forC 16 H 13 N3O2279.1008 found 279.1080.

[0148] Synthesis of compound 31 (4-(4-methoxyphenyl)-1-(3-phenylpropyl)-1H-1,2,3-triazole)

[0149] The compound bromopropylbenzene (200 mg, 1 mmol) was reacted with sodium azide in a round-bottom flask to generate an azide. It was then reacted with 1-ethynyl-4-methoxybenzene, extracted with CH2Cl2 / saturated NaHCO3, and the resulting organic layer was concentrated under reduced pressure and separated by column chromatography to give 77 mg of a white solid, with a yield of 26%.

[0150] 1H NMR (600MHz, Chloroform-d) δ7.76 (d, J=8.7Hz, 2H), 7.64 (s, 1H), 7.31 (t, J=7.6Hz, 2H), 7.23 (d, J=7.5Hz, 1H), 7.22- 7.18 (m, 2H), 6.97 (d, J=8.7Hz, 2H), 4.39 (t, J=7.1Hz, 2H), 3.85 (s, 3H), 2.70 (t, J=7.5Hz, 2H), 2.30 (p, J=7.3Hz, 2H). 13 C NMR (151MHz, Chloroform-d) δ159.60, 147.65, 140.19, 128.65, 128.49, 127.02, 126.38, 123.38, 118.75, 114.26, 55.33, 49.55, 32.53, 31.71.[M+H] + calcd for C 18 H 19 N3O 293.3700found293.1238.

[0151] Synthesis of compound 32 (4-(1-(3-phenylpropyl)-1H-1,2,3-triazol-4-yl)phenol)

[0152] Compound 31 (200 mg, 0.68 mmol) was added to a round-bottom flask, 6 mL of HBr was added, and the mixture was protected with N2. The mixture was refluxed at 120 °C and monitored by thin-layer chromatography. After 6 h, the reaction was stopped, quenched with water, and extracted with CH2Cl2 / saturated NaHCO3. The resulting organic layer was concentrated under reduced pressure and separated by column chromatography to obtain 171 mg of white solid, with a yield of 90%.

[0153] 1 H NMR (600MHz, Acetone-d6) δ8.46 (s, 1H), 8.21 (s, 1H), 7.73 (d, J=8.6Hz, 2H), 7.30 (t, J=7.5Hz, 2H), 7.28-7.24 (m, 2H), 7.22-7.18 (m, 1H), 6.90 (d, J=8.6Hz, 2H), 4.46 (t, J=7.1Hz, 2H), 2.73-2.67 (m, 2H), 2.32-2.24 (m, 2H). 13C NMR (151MHz, Acetone-d6) δ159.05, 149.03, 142.85, 130.24, 130.21, 128.55, 127.82, 124.93, 120.97, 117.32, 50.96, 34.10, 33.66.

[0154] Synthesis of compound 33 (1-(4-methoxyphenyl)-2-(4-(4-methoxyphenyl)-1H-1,2,3-triazol-1-yl)ethan-1-ol)

[0155] Compound 1 (200 mg, 0.62 mmol) and sodium borohydride and tetrahydrofuran were added to a flask and reacted at room temperature. After the reaction was completed, the mixture was quenched with water and extracted with CH2Cl2 / saturated NaHCO3. The resulting organic layer was concentrated under reduced pressure and separated by column chromatography to give 135 mg of white solid, with a yield of 67%.

[0156] 1 H NMR (600MHz, Acetone-d6) δ8.17 (s, 1H), 7.88-7.73 (m, 2H), 7.45-7.26 (m, 2H), 7.12-6.97 ( m, 2H), 6.97-6.86 (m, 2H), 5.20-5.02 (m, 1H), 4.70-4.40 (m, 2H), 3.83 (s, 3H), 3.79 (s, 3H). 13 C NMR (101MHz, Acetone-d6) δ161.30, 148.31, 135.69, 129.07, 128.39, 126.03, 122.29, 115.91, 115.49, 73.86, 58.98, 56.49, 56.43.[M+H] + calcd forC 18 H 19 N3O3325.1426 found 325.1495.

[0157] Synthesis of compound 34 (4-(1-hydroxy-2-(4-(4-hydroxyphenyl)-1H-1,2,3-triazol-1-yl)ethyl)phenol)

[0158] Compound 2 (200 mg, 0.68 mmol) and sodium borohydride and tetrahydrofuran were added to a flask and reacted at room temperature. After the reaction was completed, the mixture was quenched with water and extracted with CH2Cl2 / saturated NaHCO3. The resulting organic layer was concentrated under reduced pressure and separated by column chromatography to give 91 mg of white solid, with a yield of 45%.

[0159] 1 H NMR (400MHz, DMSO-d6) δ9.54 (s, 1H), 9.35 (s, 1H), 8.27 (s, 1H), 7.70-7.56 (m, 2H), 7.18 (d, J=8.4Hz, 2H) , 6.87-6.79 (m, 2H), 6.72 (d, J=8.5Hz, 2H), 5.65 (d, J=4.6Hz, 1H), 4.99-4.83 (m, 1H), 4.51-4.25 (m, 2H). 13 C NMR (151MHz, DMSO-d6) δ157.61, 157.29, 146.65, 132.85, 127.73, 126.95, 122.46, 121.13, 116.08, 115.42, 71.70, 57.28.[M+H] + calcd for C 16 H 15 N3O3297.1113 found 297.1184.

[0160] Synthesis of compound 35 (1-(4-methoxyphenyl)-2-(4-(3-methoxyphenyl)-1H-1,2,3-triazol-1-yl)ethan-1-ol)

[0161] Compound 3 (200 mg, 0.62 mmol) and sodium borohydride and tetrahydrofuran were added to a flask and reacted at room temperature. After the reaction was completed, the mixture was quenched with water and extracted with CH2Cl2 / saturated NaHCO3. The resulting organic layer was concentrated under reduced pressure and separated by column chromatography to give 78 mg of white solid, with a yield of 39%.

[0162] 1 H NMR (400MHz, DMSO-d6) ) δ8.52 (s, 1H), 7.48-7.25 (m, 5H), 7.06-6.75 (m, 3H), 5.75 (d, J=4.7Hz, 1H), 4.99 (dt, J=8.7, 4.5Hz, 1H), 4.49 (qd, J=13.7, 6.3Hz, 2H), 3.81 (s, 3H), 3.74 (s, 3H). 13C NMR (151MHz, DMSO-d6) δ160.15, 159.19, 146.27, 134.50, 132.74, 130.53, 127. 76, 122.84, 117.88, 114.10, 113.99, 110.65, 71.47, 57.32, 55.57, 55.52.[M+H] + calcd for C 18 H 19 N3O3325.1426 found 325.1502.

[0163] Synthesis of compound 36 (3-(1-(2-hydroxy-2-(4-hydroxyphenyl)ethyl)-1H-1,2,3-triazol-4-yl)phenol)

[0164] Compound 4 (200 mg, 0.68 mmol) and sodium borohydride and tetrahydrofuran were added to a flask and reacted at room temperature. After the reaction was completed, the mixture was quenched with water and extracted with CH2Cl2 / saturated NaHCO3. The resulting organic layer was concentrated under reduced pressure and separated by column chromatography to give 85 mg of white solid, with a yield of 42%.

[0165] 1 H NMR (400MHz, DMSO-d6) δ9.54 (s, 1H), 9.38 (s, 1H), 8.40 (s, 1H), 7.33-7.13 (m, 5H), 6.73 (dd , J=6.6, 4.6Hz, 3H), 5.68 (d, J=4.6Hz, 1H), 4.93 (dt, J=8.8, 4.6Hz, 1H), 4.54-4.36 (m, 2H). 13 C NMR (101MHz, DMSO-d6) δ158.23, 157.31, 146.44, 132.80, 132.62, 130.39, 127.73, 122.40, 116.47, 115.44, 115.24, 112.30, 71.66, 57.31.[M+H] + calcd forC 16 H 15 N3O3 297.3140 found 297.1184.

[0166] Synthesis of compound 37 (2-(4-(4-fluorophenyl)-1H-1,2,3-triazol-1-yl)-1-(4-methoxyphenyl)ethan-1-ol)

[0167] Compound 6 (200 mg, 0.64 mmol) and sodium borohydride and tetrahydrofuran were added to a flask and reacted at room temperature. After the reaction was completed, the mixture was quenched with water and extracted with CH2Cl2 / saturated NaHCO3. The resulting organic layer was concentrated under reduced pressure and separated by column chromatography to give 107 mg of white solid, yield 52%.

[0168] 1 H NMR (600MHz, Acetone-d6) δ8.30 (s, 1H), 7.98-7.88 (m, 2H), 7.45-7.35 (m, 2H), 7.27-7.1 4(m, 2H), 7.00-6.88(m, 2H), 5.15(dd, J=8.6, 4.0Hz, 1H), 4.70-4.45(m, 2H), 3.79(s, 3H). 13 C NMR (151MHz, Acetone-d6) δ164.92, 163.30, 161.29, 147.46, 135.59, 129.90, 129.88, 129.07, 123.15, 117.39, 117.24, 115.49, 56.42.[M+H] + calcd forC 16 H 16 FN3O2313.1227 found 313.1303.

[0169] Synthesis of compound 38 (1-(4-methoxyphenyl)-2-(4-phenyl-1H-1,2,3-triazol-1-y1)ethan-1-ol)

[0170] Compound 7 (200 mg, 0.67 mmol) and sodium borohydride and tetrahydrofuran were added to a flask and reacted at room temperature. After the reaction was completed, the mixture was quenched with water and extracted with CH2Cl2 / saturated NaHCO3. The resulting organic layer was concentrated under reduced pressure and separated by column chromatography to give 109 mg of white solid, with a yield of 55%.

[0171] 1 H NMR (600MHz, Acetone-d6) δ8.30 (s, 1H), 8.00-7.79 (m, 2H), 7.50-7.34 (m, 4H), 7.34-7.27 ( m, 1H), 7.07-6.83 (m, 2H), 5.15 (td, J=7.2, 6.1, 3.2Hz, 1H), 4.70-4.32 (m, 2H), 3.79 (s, 3H). 13C NMR (151MHz, Acetone-d6) δ161.28, 148.36, 135.63, 133.42, 130.53, 129.38, 129.07, 127.08, 123.23, 115.49, 73.82, 56.42.[M+H] + calcd forC 17 H 17 N3O2 295.1321 found 295.1397.

[0172] Synthesis of compound 39 (2-(4-(4-((3r,5r,7r)-adamantan-1-yl)phenyl)-1H-1,2,3-triazol-1-yl)-1-(4-methoxyphenyl)ethan-1-one)

[0173] 1-Hydroxyadamantane (200 mg, 1.32 mmol) was added to a round-bottom flask with bromobenzene, trifluoromethanesulfonic acid, and CH₂Cl₂. The reaction was carried out at 0 °C, quenched with water after completion, and extracted with saturated NaHCO₃ CH₂Cl₂. The resulting organic layer was concentrated under reduced pressure and separated by column chromatography to obtain a white solid. The obtained compound was reacted with trimethylsilaneacetylene in DMF and Et₃N in the presence of PdCl₂(PPh₃)₂ and CuI. Then, the trimethylsilyl group was removed, and the compound was reacted with an azide to give 62 mg of the product, with a yield of 11%.

[0174] 1 H NMR (600MHz, Chloroform-d) δ8.01 (d, J=8.9Hz, 2H), 7.91 (s, 1H), 7.81 (d, J=8.4Hz, 2H), 7.43 (d, J=8.4Hz, 2H), 7.01 (d , J=8.9Hz, 2H), 5.83 (s, 2H), 3.91 (s, 3H), 2.11 (s, 3H), 1.95 (d, J=2.9Hz, 6H), 1.79 (q, J=12.8Hz, 6H).HRMS (m / z) [M+H] + calcd forC 27 H 29 N3O2428.2260found 428.2336.

[0175] Synthesis of compound 40 (2-(4-(4-((3r,5r,7r)-adamantan-1-yl)phenyl)-1H-1,2,3-triazol-1-yl)-1-(4-hydroxyphenyl)ethan-1-one)

[0176] Compound 39 (200 mg, 0.47 mmol) was added to a round-bottom flask, 6 mL of HBr was added, N2 protection was applied, and the mixture was refluxed at 120 °C. Thin-layer chromatography was used for monitoring. After 6 h, the reaction was stopped, quenched with water, and extracted with CH2Cl2 / saturated NaHCO3. The resulting organic layer was concentrated under reduced pressure and separated by column chromatography to obtain 45 mg of white solid, with a yield of 23%.

[0177] 1 H NMR (600MHz, DMSO-d6) δ10.59 (s, 1H), 8.44 (s, 1H), 7.97 (d, J=8.7Hz, 2H), 7.79 (d, J=8.4Hz, 2H), 7.45 (d, J=8. 5Hz, 2H), 6.93 (d, J=8.7Hz, 2H), 6.10 (s, 2H), 2.08 (s, 3H), 1.90 (d, J=2.9Hz, 6H), 1.75 (s, 6H).HRMS (m / z) [MH] - calcd forC 26 H 27 N3O2412.2103 found 412.2035.

[0178] Example 3: HIF-1 Activity Inhibition Experiment

[0179] S1 Material Preparation

[0180] Cell culture in 96-well plates, high-glucose medium, fetal bovine serum, 10× phosphate buffer, 0.05% trypsin solution, cell lysis buffer, and dual-luciferase reporter gene assay kit.

[0181] S2 Preparation of Cell Suspension

[0182] Discard the culture medium from the cultured, stably transfected HeLa cells, wash 1-2 times with PBS, add trypsin, digest in an incubator for 60 seconds, discard the trypsin, add culture medium, mix the cell suspension thoroughly, count the cells, dilute the cell suspension to 200,000 / ml.

[0183] S3 seeded cells

[0184] 100 μL of cell suspension was seeded into 96-well white plates and placed in a normal oxygen incubator for 12 h.

[0185] S4 dosing

[0186] The compound was diluted with culture medium and added to the corresponding air. The control group was added with the corresponding volume of culture medium. The normoxic and hypoxic plates were placed in the corresponding incubators for 12 hours.

[0187] S5 Test

[0188] The prepared cell lysis buffer was added to the wells of the plate and the cells were lysed. The cells were then tested using a dual-luciferase reporter gene assay kit.

[0189] The experimental results are shown in Table 1.

[0190] Example 4 Cytotoxicity Test

[0191] S1 Material Preparation

[0192] Cell culture in 96-well plates, high-glucose medium, fetal bovine serum, 10× phosphate buffer, and 0.05% trypsin solution.

[0193] S2 Preparation of Cell Suspension

[0194] Discard the culture medium after culture, wash the cells 1-2 times with PBS, add trypsin, digest in an incubator for 60 seconds, discard the trypsin, add an appropriate amount of complete culture medium, mix the cell suspension thoroughly, count the cells, dilute the cell suspension to 20,000 cells / ml.

[0195] S3 seeded cells

[0196] Prepare a 96-well plate, add 100 μl of cell suspension to each well, and incubate in an incubator (37℃, 5% CO2) for 48 h.

[0197] S4 Testing Method

[0198] After 48 hours, the liquid in the wells was aspirated, and 200 μl of DMSO was added to each well. After shaking the plate for 10 minutes, the OD value of each well was measured at a wavelength of 570 nm.

[0199] The experimental results are shown in Table 1.

[0200] Example 5: Cancer Cell Invasion Inhibition Experiment

[0201] S1 Material Preparation

[0202] Transwell chamber, bright-field microscope with photography capability, 24-well cell culture plates, high-glucose medium, fetal bovine serum, 10× phosphate buffer, 0.05% trypsin solution, methanol, crystal violet staining solution, Matrigel.

[0203] S2 lays Matrigel

[0204] Thaw Matrigel from -20°C to 4°C 12 hours in advance. After 12 hours, dilute Matrigel with DMEM (Matrigel∶DMEM=1∶5) and mix well. Spread 50 μL of the diluted Matrigel evenly in a chamber and incubate at 37°C for 30 min to allow it to solidify.

[0205] S3 Preparation of Cell Suspension

[0206] Discard the culture medium after culture, wash the cells 1-2 times with PBS, add trypsin, digest in an incubator for 60 seconds, discard the trypsin, add an appropriate amount of low-serum culture medium (high-glucose medium containing 2% fetal bovine serum), centrifuge, discard the supernatant, add low-serum culture medium again, resuspend the cells evenly, count the cells, dilute the cells, and adjust the cell suspension to 400,000 / ml.

[0207] S4 seeded cells

[0208] Remove the chamber, add 650 μl of complete culture medium to the lower chamber, and add 100 μl of cell suspension to the upper chamber. Ensuring there are no air bubbles in either chamber, place the upper chamber back into the 24-well plate.

[0209] S5 dosing

[0210] The compound was diluted with low-serum culture medium, and 100 μl was added to the upper chamber. The cell culture plate was then returned to the incubator for 24 hours.

[0211] S6 Result Processing

[0212] Remove the chamber, discard the culture medium, wash three times with 1×PBS, fix with methanol for 20 min, then wash three more times with 1×PBS, and stain with crystal violet for 20 min. Gently rinse away any excess crystal violet with water, wipe away the upper layer of cells with a cotton swab, and photograph under a bright-field microscope.

[0213] Based on the experimental results ( Figure 1 Compound 10a (18) can inhibit the invasion of cancer cells at low micromolar concentrations.

[0214] Example 6: HUVEC Cell Migration Inhibition Experiment

[0215] S1 Material Preparation

[0216] Cell culture 6-well plates, photographic bright-field microscope, high-glucose culture medium, fetal bovine serum, 10× phosphate buffer solution, 0.05% trypsin solution

[0217] S2 Preparation of Cell Suspension

[0218] Discard the culture medium after culture, wash the cells 1-2 times with PBS, add trypsin, digest for 60 seconds in an incubator, discard the trypsin, add an appropriate amount of complete culture medium, mix the cell suspension thoroughly, count the cells, dilute the cell suspension to 500,000 / ml.

[0219] S3 seeded cells

[0220] Prepare a six-well plate, draw three parallel lines on the bottom of the plate with a marker, add 1 ml of cell suspension to each well, and incubate in an incubator.

[0221] S4 scratches

[0222] Once the cells have spread evenly, use a 200 μl pipette tip to draw lines vertically from the bottom. Wash 2-3 times with 1×PBS, then add 1 ml of low-serum culture medium (high-glucose medium containing 2% fetal bovine serum).

[0223] S5 Result Processing

[0224] At 0h, three random fields of view were photographed under a bright-field microscope, and photographs were taken again 24h later. The results were processed using ImageJ.

[0225] Based on the experimental results ( Figure 2 Compound 10a (18) can inhibit cancer cell migration in a concentration-dependent manner at low micromolar concentrations.

[0226] Example 7: Cancer Cell Clonal Proliferation Inhibition Experiment

[0227] S1 Material Preparation

[0228] Cell culture 6-well plates, photographic bright-field microscope, high-glucose culture medium, fetal bovine serum, 10× phosphate buffer solution, 0.05% trypsin solution, methanol, crystal violet staining solution.

[0229] S2 Preparation of Cell Suspension

[0230] Discard the culture medium after culture, wash the cells 1-2 times with PBS, add trypsin, digest in an incubator for 60 seconds, discard the trypsin, add an appropriate amount of complete culture medium, mix the cell suspension thoroughly, count the cells, dilute the cell suspension to 1000 cells / ml.

[0231] S3 seeded cells

[0232] Prepare a six-well plate, add 1 ml of cell suspension to each well, and incubate in a hypoxic incubator (1% O2).

[0233] S4 dosing

[0234] 24 hours after inoculation, administer the drug at the pre-designed concentration. Subsequently, on days 3, 6, and 9 after drug administration, discard the culture medium and administer the drug again.

[0235] S5 Result Processing

[0236] Remove the six-well plate, discard the culture medium, wash three times with 1×PBS, fix with methanol for 20 min, then wash three more times with 1×PBS, and stain with crystal violet for 20 min. Gently rinse off any excess crystal violet with water, allow to air dry, and then photograph.

[0237] Based on the experimental results ( Figure 3Compound 10a (18) can inhibit the clonal proliferation of cancer cells at low micromolar concentrations.

[0238] Example 8 Real-time quantitative PCR

[0239] S1 Material Preparation

[0240] Low-temperature high-speed centrifuge; reverse transcription apparatus; PCR amplification apparatus; centrifuge mixer; TRIzol reagent; chloroform; isopropanol; ethanol; DEPC water; ultrapure water; reverse transcription kit; PCR amplification kit

[0241] S2 type board

[0242] HeLa cells were seeded into six-well plates at 400,000 cells per well and cultured for 24 hours.

[0243] S3RNA quantification

[0244] Cells were lysed, separated into layers using chloroform, precipitated with isopropanol, washed with 75% ethanol, and the resulting RNA was quantified.

[0245] S4 reverse transcription

[0246] RNA was reverse transcribed using a reverse transcription kit following the prescribed steps.

[0247] S5 Real-time Quantitative PCR Amplification

[0248] Add primers according to the kit instructions for amplification.

[0249] S6 detection

[0250] Set up the program, perform the test, and analyze the results.

[0251] Based on the experimental results ( Figure 4 Compound 10a (18) can inhibit the expression of VEGF mRNA.

[0252] Example 9: Surface Plasma Resonance Experiment

[0253] S1 Material Preparation

[0254] CM5 chip, Hsp90 protein, sodium acetate, protein conjugation kit, NaOH solution, run buffer, Tween 20

[0255] S2 protein coupling

[0256] The protein was pre-enriched to determine the pH required for formal coupling, and then formal coupling was performed. After the target coupling amount was reached, the chip was sealed.

[0257] S3 Affinity Test

[0258] Prepare samples of different concentrations of small molecules for testing, set the corresponding program after loading them onto the instrument, and perform the tests.

[0259] S4 software simulation

[0260] The test results were simulated using software to obtain affinity data between small molecules and proteins.

[0261] Based on the experimental results ( Figure 5 The KD of compound 10a is 3.72 μM, thus Hsp90 is determined to be the target of compound 10a.

[0262] Example 10 Molecular docking

[0263] S1 small molecule preparation

[0264] In Discovery Studio, draw the 10a structure, add hydrogen and minimize the energy of the drawn structure, and select Prepare Ligands in Prepare or Filter Ligands under the Small Molecules module to prepare the small molecule for docking.

[0265] S2 protein molecule processing

[0266] Download the 3EKR protein structure from the PDB database using Discovery Studio software. First, delete the protein B chain, then delete the bound water molecules except for the N-terminal ATP pocket. Select PrepareProtein under the Macromolecules module to process the protein molecule and define the docking receptor and binding site.

[0267] S3 Cdocker Molecular Docking

[0268] Expand Receptor-Ligand Interactions, select Dock Ligands (CDOCKER) under Dock Ligands, set the corresponding receptor and the processed small molecule, select the coordinates and radius of the docking site from the drop-down list, click Run to perform docking, and view the analysis results after the process is complete.

[0269] Appendix Figure 6 This diagram illustrates the interaction between compound 10a and Hsp90. As shown, compound 10a forms a key hydrogen bond with asparagine at position 51. The triazole moiety forms hydrogen bonds with glycine at position 97 and threonine at position 184. The amino and triazole moieties of 10a can form hydrogen bonds with bound water at positions 902 and 903, and the carbonyl group of 10a acts as a hydrogen bond acceptor, forming a hydrogen bond with asparagine at position 106 of Hsp90.

[0270] AppendixFigure 7 This is a schematic diagram illustrating the interaction between compound 33 and Hsp90. The triazole ring and hydroxyl group simultaneously form key hydrogen bonds with Lys58. Meanwhile, the methoxy group on the benzene ring forms a hydrogen bond with a bound water molecule and Thr184, respectively, while the other side of the benzene ring exhibits hydrophobic interactions with Lys112.

[0271] Example 11: In vivo antitumor activity study

[0272] S1 Breast Cancer Mouse Model Construction

[0273] 4T1 cells were digested, resuspended in PBS, and the cell density was adjusted to 3 × 10⁻⁶ cells / mL. 5 The cells were seeded under the second pair of mammary pads in BALB / c (6-7 weeks) mice, and tumors formed after 7 days, indicating successful model construction.

[0274] S2 mice grouping and administration

[0275] Mice with tumors were divided into four groups: 1. Negative control group; 2. Cisplatin (DDP) group; 3. Single-drug group; 4. Combined-drug group. Cisplatin was administered every 5 days at a dose of 2 mg / kg via intraperitoneal injection, and for 10 years, it was administered daily at a dose of 30 mg / kg via tail vein injection. The weight and tumor size of the mice were measured and recorded daily.

[0276] S3 Tumor Suppression Effect Evaluation

[0277] Thirteen days after administration, the mice were sacrificed, and the tumors of each group of mice were removed, weighed, and photographed.

[0278] Based on the experimental results ( Figures 8-10 Compound 10a can effectively inhibit tumor growth. When used in combination with cisplatin, it can inhibit tumor growth even more effectively, and the body weight of mice did not change significantly.

[0279] Table 1. Data on the toxicity of compounds to cancer cells and their inhibitory activity against HIF-1.

[0280]

[0281]

[0282] The structures of compounds 10a and 4f in the above embodiments are as follows:

[0283]

[0284] The above are merely preferred embodiments of the present invention and are not intended to limit the present invention. Any modifications, equivalent substitutions, improvements, etc., made within the scope of the concept and principles of the present invention should be included within the protection scope of the present invention.

Claims

1. The use of a compound in the preparation of a drug for treating breast cancer, characterized in that, The structure of the compound: 。 2. The application according to claim 1, characterized in that, The drugs are tablets, capsules, pills, injections, drops, granules, suppositories, aerosols, sprays, powder inhalers, syrups, tinctures, lotions, and films.

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