Izoxazole-containing azole derivatives, processes for their preparation and use

Abstract

The application belongs to the technical field of medicine, and relates to an isoxazole-containing azole derivative and a preparation method and medical application thereof. The compound is shown as general formula (1). The compound realizes strong and broad-spectrum antifungal activity, and has potential application as an antifungal drug. The definition of R in the general formula (1) is shown in the specification.

Description

Technical Field

[0001] This invention belongs to the field of pharmaceutical technology and relates to azole derivatives containing isoxazole structures, their preparation methods, and their pharmaceutical uses. Background Technology

[0002] Invasive fungal infections (IFIs) are becoming a major infectious disease worldwide. Among existing antifungal drugs, CYP51 inhibitors are the most widely used in clinical practice due to their significant antifungal effects. Azole drugs are cytochrome P450-14α-demethylase (CYP51) inhibitors, which prevent the biosynthesis of ergosterol in fungi, leading to changes in cell membrane permeability and thus inhibiting fungal growth.

[0003] It has been reported that the CYP51 active site has three distinct parts: (a) an azole ring (coordinated with the heme iron of CYP51); (b) a hydrophobic channel I; and (c) another narrow hydrophobic channel II (substrate entry channel). Currently, clinically used azole antifungal drugs mainly include fluconazole, miconazole, voriconazole, and ivorconazole. The hepatotoxicity of azole drugs manifests as widespread drug-drug interactions, primarily due to their interaction with human CYP enzymes, limiting the clinical application of azole drugs. On the other hand, most azole drugs are excessively hydrophobic, resulting in poor adsorption-metabolism (ADME) properties. Optimizing the ADME properties of azole drugs is currently a direction for developing novel azole drugs. Summary of the Invention

[0004] The purpose of this invention is to design and synthesize azole derivatives containing isoxazole structures, their preparation methods, and their pharmaceutical applications.

[0005] To achieve the above objectives, the present invention adopts the following technical solution:

[0006] An azole derivative containing an isoxazole structure, the compound is shown in general formula (1).

[0007]

[0008] In the general formula:

[0009] X, Y, and Z can be the same or different and are selected from N or CH;

[0010] R 1 R 2 R 3 R 4The following groups may be selected from hydrogen, halogen, cyano, nitro, hydroxyl, and amino groups, either unsubstituted or substituted with at least one of the following groups: C1-6 alkyl, C3-8 cycloalkyl, C2-6 alkenyl, C2-6 alkoxy, C2-6 alkenyloxy, and C2-6 alkynoxy. These groups may be selected from halogen, hydroxyl, cyano, nitro, and amino groups, either identically or differently. The following amino groups may be unsubstituted or substituted with groups such as those selected from C1-6 alkyl, C1-6 aminoalkyl, C1-6 hydroxyalkyl, C1-6 alkoxyalkyl, and C1-6 cyanoalkyl. C2-6 alkenyl, C2-6 alkynyl, C1-6 alkylsulfonyl, C1-6 alkyl carbonyl, C2-6 alkenyl carbonyl or C2-6 alkynyl carbonyl; unsubstituted or substituted thioamide or amide group, wherein the substituted position of the thioamide or amide group is amino, and the substituted group is selected from C1-6 alkyl, C1-6 aminoalkyl, C1-6 hydroxyalkyl, C1-6 alkoxyalkyl, C1-6 cyanoalkyl, C2-6 alkenyl, C2-6 alkynyl, C1-6 alkylsulfonyl, C1-6 alkyl carbonyl, C2-6 alkenyl carbonyl or C2-6 alkynyl carbonyl;

[0011] Or, the optical isomers of the compound represented by general formula (1), or diastereomers.

[0012] Preferably, in the general formula:

[0013] X, Y, and Z can be the same or different and are selected from N or CH;

[0014] R 1 R 2 R 3 R 4 The same or different groups may be selected from hydrogen atoms, halogens, cyano, nitro, hydroxyl, amino, unsubstituted or substituted by at least one of the following groups: C1-6 alkyl, C3-8 cycloalkyl, C2-6 alkenyl, C2-6 alkoxy, C2-6 alkenyloxy, C2-6 alkoxy, C2-6 alkoxy, C2-6 alkoxy, and the following groups may be selected from halogens, hydroxyl, cyano, nitro, and amino;

[0015] Or, the optical isomers of the compound represented by general formula (1), or diastereomers.

[0016] Further preferred, in the general formula:

[0017] X, Y, and Z can be the same or different and are selected from N or CH;

[0018] R 1 R 2 R 3 R 4The same or different groups may be selected from hydrogen atoms, halogens, cyano, nitro, hydroxyl, amino, unsubstituted or substituted by at least one of the following groups, which may be selected from halogens, hydroxyl, cyano, nitro and amino;

[0019] Or, the optical isomers of the compound represented by general formula (1), or diastereomers.

[0020] Further preferred, in the general formula:

[0021] X, Y, and Z can be the same or different and are selected from N or CH;

[0022] R 1 R 2 R 3 R 4 The same or different groups may be selected from hydrogen atoms, halogens, cyano, nitro, hydroxyl, amino, unsubstituted or substituted by at least one of the following groups, C1-4 alkyl, C1-4 alkoxy, which may be the same or different groups selected from halogens or hydroxyl;

[0023] Or, the optical isomers of the compound represented by general formula (1), or diastereomers.

[0024] Furthermore, in the general formula:

[0025] X, Y, and Z can be the same or different and are selected from N or CH;

[0026] R 1 R 2 R 3 R 4 The same or different groups may be selected from hydrogen atoms, halogens, cyano, nitro, hydroxyl, amino, unsubstituted or substituted by at least one of the following groups, C1-4 alkyl, C1-4 alkoxy, which may be the same or different groups selected from halogens;

[0027] Or, the optical isomers of the compound represented by general formula (1), or diastereomers.

[0028] The most preferred compounds represented by general formula (1) are the following E01-E14 compounds and their optical isomers, diastereomers;

[0029]

[0030] A method for preparing the compound is described below: First, acetophenone containing different substituents on the benzene ring reacts with diethyl oxalate to generate intermediate Z-1a-f, then reacts with hydroxylamine hydrochloride to generate isoxazole ring intermediate Z-2a-f, which is then reduced and brominated to obtain intermediate Z-4a-f. Compounds a and b undergo substitution reactions with p-bromobenzyl bromide to generate intermediate Z-5a-b, which is then boronized and reacted with intermediate Z-4a-f via a Suzuki reaction to obtain the compound of general formula (1).

[0031]

[0032] A pharmaceutical composition comprising the said compound and its optically active form, diastereomer, and pharmaceutically acceptable carrier.

[0033] The use of a compound and composition, the use of said compound and its optically active form, diastereomer, or said composition in the preparation of antifungal drugs. More specifically, its use in the preparation of drugs that inhibit CYP51.

[0034] The use of the compound and its optically active form, diastereomer, or the composition of claim 7 in the preparation of antifungal drugs.

[0035] Advantages of this invention:

[0036] The compounds of the present invention are designed based on the active cavity of CYP51 to obtain the compounds described in general formula (1). These compounds have a azole functional group that coordinates with heme iron porphyrin, an aromatic ring region, and a long chain segment that can extend into the narrow protein cavity, thereby achieving the inhibitory activity of CYP51 and improving the antifungal activity. They have potential applications as antifungal drugs. Detailed Implementation

[0037] The following examples will provide a better understanding of the compounds of the present invention and their preparation. These examples are intended to illustrate, rather than limit, the scope of the invention.

[0038] Example 1: Ethyl 2,4-dioxo-4-phenylbutyrate (Z-1a)

[0039] In a 500 mL round-bottom flask, acetophenone (10 g, 83 mmol), diethyl oxalate (18.24 g, 120 mmol, 1.4 eq), sodium hydride (6.66 g, 208 mmol, 2.5 eq), and tetrahydrofuran (40 mL) were added. The mixture was heated at 0 °C for 6 h with stirring. 100 mL of water was added to the reaction mixture, and the mixture was extracted with ethyl acetate (3 × 100 mL). The combined organic phases were washed successively with water (3 × 50 mL) and saturated brine (50 mL). The mixture was dried over anhydrous sodium sulfate, filtered through the drying agent, and concentrated under reduced pressure to obtain the crude product Z-1a as a yellow solid, yielding 13.2 g. The crude product was not purified and proceeded directly to the next reaction. HRMS (ESI): Calcd.for C 12 H 12 O4[M+Na] + :243.0628,Found 243.0638[M+Na] + .

[0040] Ethyl 4-(4-fluorophenyl)-2,4-dioxobutyrate (Z-1b)

[0041] Using 4-fluoroacetophenone as a starting material, a white solid was obtained following the synthetic method of intermediate Z-1a. HRMS(ESI): Calcd.for C 12 H 11 FO4[M+Na] + :261.0534,Found 261.0540[M+Na] + .

[0042] Ethyl 4-(2,4-difluorophenyl)-2,4-dioxobutyrate (Z-1c)

[0043] Using 2,4-difluoroacetophenone as a starting material, and following the synthetic method for intermediate Z-1a, a white solid, ethyl 4-(4-trifluoromethylphenyl)-2,4-dioxobutyrate (Z-1d), was obtained.

[0044] Using 4-trifluoromethylacetophenone as a raw material, a white solid was obtained by referring to the synthesis method of intermediate Z-1a.

[0045] Ethyl 4-(4-methylphenyl)-2,4-dioxobutyrate (Z-1e)

[0046] Using 4-methylacetophenone as a starting material, and following the synthetic method for intermediate Z-1a, a white solid, ethyl 4-(4-methoxyphenyl)-2,4-dioxobutyrate (Z-1f), was obtained.

[0047] Using 4-methoxyacetophenone as a starting material, and following the synthetic method for intermediate Z-1a, a white solid was obtained. HRMS(ESI): Calcd.for C13 H 14 O5[M+Na] + :273.0733,Found 273.0739[M+Na] + .

[0048] Example 2: Ethyl 5-phenylisoxazole-3-carboxylate (Z-2a)

[0049] In a 250 mL round-bottom flask, intermediate Z-1a (13.2 g, 60 mmol), hydroxylamine hydrochloride (8.34 g, 120 mmol), and 60 mL of ethanol were added. The mixture was reacted at 85 °C for 4 h with stirring. Part of the solvent was removed by vacuum distillation, 40 mL of water was added, and the mixture was filtered to obtain a solid. The solid was dried and purified by column chromatography (petroleum ether: ethyl acetate = 10:1) to give Z-2a as a white solid, with a yield of 7.5 g (57.65%).

[0050] 5-(4-fluorophenyl)isoxazole-3-carboxylic acid ethyl ester (Z-2b)

[0051] Using intermediate Z-1b as a raw material and following the synthesis method of intermediate Z-2a, a white solid was obtained. HRMS(ESI): Calcd.for C 12 H 10 FNO3[M+Na] + :258.0537,Found 258.0537[M+Na] + .

[0052] 5-(2,4-Difluorophenyl)isoxazole-3-carboxylic acid ethyl ester (Z-2c)

[0053] Using intermediate Z-1c as raw material and referring to the synthesis method of intermediate Z-2a, a white solid was obtained.

[0054] 5-(4-Trifluoromethylphenyl)isoxazole-3-carboxylic acid ethyl ester (Z-2d)

[0055] Using intermediate Z-1c as a starting material and following the synthesis method of intermediate Z-2a, a white solid was obtained. HRMS(ESI): Calcd.for C 13 H 10 F3NO3[M+Na] + :308.0505,Found 308.0521[M+Na] + .

[0056] 5-(4-Methylphenyl)isoxazole-3-carboxylic acid ethyl ester (Z-2e)

[0057] Using intermediate Z-1e as a starting material and following the synthesis method of intermediate Z-2a, a white solid was obtained. HRMS(ESI): Calcd.for C 13 H 13 NO3[M+Na] + :254.0788,Found 254.0792[M+Na] + .

[0058] 5-(4-Methoxyphenyl)isoxazole-3-carboxylic acid ethyl ester (Z-2f)

[0059] Using intermediate Z-1f as a raw material and following the synthesis method of intermediate Z-2a, a white solid was obtained. HRMS(ESI): Calcd.for C 13 H 13 NO4[M+Na] + :270.0737,Found 270.0748[M+Na] + .

[0060] Example 3: (5-Phenylenoxazol-3-yl)methanol (Z-3a)

[0061] In a 250 mL round-bottom flask, intermediate Z-2a (7.5 g, 35 mmol) and 60 mL of methanol were added, followed by the addition of sodium borohydride (5.12 g, 138 mmol) in portions. The mixture was reacted at 25 °C for 2 h with stirring. Part of the solvent was removed by vacuum distillation, 40 mL of water was added, and the mixture was filtered to obtain a solid. After drying, Z-3a was given as a white solid, with a yield of 5.93 g (98.02%). HRMS (ESI): Calcd.for C 10 H9NO2[M+Na] + :198.0525,Found198.0528[M+Na] + .

[0062] (5-(4-fluorophenyl)isoxazol-3-yl)methanol(Z-3b)

[0063] Using intermediate Z-2b as a starting material and following the synthetic method for intermediate Z-3a, a white solid, (5-(2,4-difluorophenyl)isoxazol-3-yl)methanol (Z-3c), was obtained.

[0064] Using intermediate Z-2c as raw material and referring to the synthesis method of intermediate Z-3a, a white solid was obtained.

[0065] (5-(4-trifluoromethylphenyl)isoxazol-3-yl)methanol(Z-3d)

[0066] Using intermediate Z-2d as a starting material and following the synthetic method for intermediate Z-3a, a white solid, (5-(4-methylphenyl)isoxazol-3-yl)methanol (Z-3e), was obtained.

[0067] Using intermediate Z-2e as a starting material and following the synthesis method of intermediate Z-3a, a white solid was obtained. HRMS(ESI): Calcd.for C 11 H 11 NO2[M+Na] + :212.0682,Found 212.0697[M+Na] + .

[0068] (5-(4-methoxyphenyl)isoxazol-3-yl)methanol (Z-3f)

[0069] Using intermediate Z-2f as a starting material and following the synthesis method of intermediate Z-3a, a white solid was obtained. HRMS(ESI): Calcd.for C 11 H 11 NO3[M+Na] + :228.0631,Found 228.0635[M+Na] + .

[0070] Example 4: 3-(bromomethyl)-5-phenylisoxazole (Z-4a)

[0071] In a 100 mL round-bottom flask, intermediate Z-3a (1.58 g, 9 mmol), carbon tetrabromide (4.48 g, 13.5 mmol), and 15 mL of dichloromethane were added. The mixture was stirred and reacted at 25 °C for 6 h. Part of the solvent was removed by vacuum distillation, 40 mL of water was added, and the mixture was extracted with dichloromethane (3 × 30 mL). The organic phases were combined and washed successively with water (3 × 20 mL), saturated brine (20 mL), and dried over anhydrous sodium sulfate. After filtration through the drying agent, the mixture was concentrated under reduced pressure to obtain the crude product. The crude product was purified by column chromatography (petroleum ether: ethyl acetate 15:1) to give Z-4a as a white solid, with a yield of 1.45 g and a recovery rate of 67.75%.

[0072] 3-(bromomethyl)-5-(4-fluorophenyl)isoxazole (Z-4b)

[0073] Using intermediate Z-3b as raw material and referring to the synthesis method of intermediate Z-4a, a white solid was obtained.

[0074] 3-(bromomethyl)-5-(2,4-difluorophenyl)isoxazole (Z-4c)

[0075] Using intermediate Z-3c as raw material and referring to the synthesis method of intermediate Z-4a, a white solid was obtained.

[0076] 3-(bromomethyl)-5-(4-trifluoromethylphenyl)isoxazole (Z-4d)

[0077] Using intermediate Z-3d as raw material and referring to the synthesis method of intermediate Z-4a, a white solid was obtained.

[0078] 3-(bromomethyl)-5-(4-methylphenyl)isoxazole (Z-4e)

[0079] Using intermediate Z-3e as raw material and referring to the synthesis method of intermediate Z-4a, a white solid was obtained.

[0080] 3-(bromomethyl)-5-(4-methoxyphenyl)isoxazole (Z-4f)

[0081] Using intermediate Z-3f as a raw material and following the synthesis method of intermediate Z-4a, a white solid was obtained. HRMS(ESI): Calcd.for C 11 H 11 NO3[M+H] + :267.9968,Found 267.9974[M+H] + .

[0082] Example 5: 1-(2-((4-bromobenzyl)oxy)-2-(2,4-difluorophenyl)ethyl)-1H-1,2,4-triazole (Z-5a)

[0083] In a 50 mL round-bottom flask, add 1-(2,4-difluorophenyl)-2-(1H-1,2,4-triazole)ethanol (0.50 g, 2.22 mmol), p-benzyl bromide (1.11 g, 4.44 mmol, 2 eq), sodium hydride (0.12 g, 5 mmol, 2.5 eq), and 5 mL of tetrahydrofuran. Heat at 70 °C for 3 h with stirring. Add 10 mL of water to the reaction mixture, extract with ethyl acetate (3 × 10 mL), combine the organic phases, wash successively with water (3 × 10 mL), wash with saturated brine (10 mL), dry with anhydrous sodium sulfate, filter through the drying agent, concentrate under reduced pressure to obtain the crude product, and purify by column chromatography (dichloromethane:methanol = 20:1) to give Z-5 as a yellow solid, yield 0.76 g, 86.40%. HRMS(ESI):Calcd.for C 17 H 14 BrF₂N₃O[M+H] + :394.0361,Found 394.0374[M+H] + .

[0084] 1-(2-((4-bromobenzyl)oxy)-2-(2,4-dichlorophenyl)ethyl)-1H-imidazolium (Z-5b)

[0085] Starting with 1-(2,4-dichlorophenyl)-2-(1H-imidazolium)ethanol, a white solid was obtained following the synthetic method of intermediate Z-5a. HRMS(ESI): Calcd.for C 17 H 14 BrCl2N3O[M+H] + :425.9770,Found 425.9766[M+H] + .

[0086] Example 6: 1-(2-(2,4-difluorophenyl)-2-((4-(4,4,5,5-tetramethyl-1,3,2-dioxaborane-2-yl)benzyloxy)ethyl)-1H-1,2,4-triazole (Z-6a)

[0087] In a 50 mL three-necked flask, intermediate Z-5 (3.7 g, 9.39 mmol, 1 eq), pinacol diboronate (3.58 g, 14.08 mmol, 1.5 eq), potassium acetate (3.38 g, 28.17 mmol, 3 eq), 1,1'-bis(diphenylphosphine)ferrocene palladium chloride (0.2 g, 0.28 mmol, 0.03 eq), and 10 mL of dioxane were added. The reaction mixture was heated at 80 °C for 4 h under nitrogen protection. 10 mL of water was added to the reaction mixture, and the mixture was extracted with ethyl acetate (3 × 10 mL). The organic phases were combined and washed successively with water (3 × 10 mL) and saturated brine (10 mL). The mixture was dried over anhydrous sodium sulfate, filtered through the drying agent, and concentrated under reduced pressure to obtain the crude product. The crude product was purified by column chromatography (dichloromethane:methanol = 20:1) to give Z-6a as a yellow solid, with a yield of 1.75 g (42.27%). HRMS(ESI):Calcd.forC 23 H 26 BF2N3O3[M+Na] + :464.1927,Found 464.1948[M+Na] + .

[0088] 1-(2-(2,4-dichlorophenyl)-2-((4-(4,4,5,5-tetramethyl-1,3,2-dioxaborane-2-yl)benzyloxy)ethyl)-1H-imidazolium (Z-6b)

[0089] Using intermediate Z-5b as a raw material and following the synthesis method of intermediate Z-6a, a yellow solid was obtained. HRMS(ESI): Calcd.for C 23 H 26 BF2N3O3[M+Na] + :497.1782,Found 497.1763[M+Na] + .

[0090] Example 7: 3-(4-((1-(2,4-difluorophenyl)-2-(1H-1,2,4-triazol-1-yl)ethoxy)methyl)benzyl)-5-phenylisoxazole (E01)

[0091] In a 50 mL three-necked flask, intermediates Z-4a (0.5 g, 2.1 mmol, 1 eq), Z-6 (0.92 g, 2.1 mmol, 1 eq), cesium carbonate (2.05 g, 6.3 mmol, 3 eq), 1,1'-bis(diphenylphosphine)ferrocene palladium chloride (0.043 g, 0.06 mmol, 0.03 eq), and 10 mL of dioxane were added. The reaction mixture was heated at 110 °C for 4 h under nitrogen protection. 10 mL of water was added to the reaction mixture, and the mixture was extracted with ethyl acetate (3 × 10 mL). The organic phases were combined and washed successively with water (3 × 10 mL), then with saturated brine (10 mL). The mixture was dried over anhydrous sodium sulfate, filtered through the drying agent, and concentrated under reduced pressure to obtain the crude product. The crude product was purified by column chromatography (dichloromethane:methanol = 20:1) to give E01 as a yellow solid, with a yield of 0.3 g and a yield of 30.6%. HRMS(ESI):Calcd.forC 27 H 22 F2N4O2[M+Na] + :495.1603,Found 495.1603[M+Na] + . 1 H NMR(400MHz,DMSO-d6)δ8.45(s,1H),7.94(s,1H),7.83(dd,J=8.0,1.7Hz,2H ),7.52-7.47(m,4H),7.31-7.26(m,1H),7.23(d,J=8.2Hz,2H),7.16-7.13(m, 1H),7.05(d,J=8.1Hz,2H),6.85(s,1H),5.06(dd,J=8.4,4.2Hz,1H),4.58(dd ,J=14.1,8.3Hz,1H),4.44-4.35(m,2H),4.22(d,J=12.0Hz,1H),4.01(s,2H).

[0092] Example 8: 3-(4-((1-(2,4-difluorophenyl)-2-(1H-1,2,4-triazol-1-yl)ethoxy)methyl)benzyl)-5-(4-fluorophenyl)isoxazole (E02)

[0093] Using intermediates Z-4b and Z-6a as raw materials, and following the synthesis method of intermediate E01, a yellow solid E02 was obtained. HRMS(ESI):Calcd.for C 27 H21 F3N4O2[M+Na] + :513.1509,Found 513.1516[M+Na] + . 1 H NMR(400MHz,DMSO-d6)δ8.45(s,1H),8.00-7.86(m,3H),7.52-7.45(m,1H),7.36-7.03(m,8H),6.85(s,1H ),5.05(dd,J=8.4,4.2Hz,1H),4.61-4.54(m,1H),4.44-4.35(m,2H),4.22(d,J=12.0Hz,1H),4.00(s,2H).

[0094] Example 9: 3-(4-((1-(2,4-difluorophenyl)-2-(1H-1,2,4-triazol-1-yl)ethoxy)methyl)benzyl)-5-(2,4-difluorophenyl)isoxazole (E03)

[0095] Using intermediates Z-4c and Z-6a as raw materials, and following the synthesis method of intermediate E01, a yellow solid E03 was obtained. HRMS(ESI): Calcd.for C 27 H 20 F4N4O2[M+Na] + :531.1415,Found 531.1422[M+Na] + . 1 H NMR(400MHz,DMSO-d6)δ8.45(s,1H),7.98-7.92(m,2H),7.52-7.46(m,2H),7.29-7.22(m,4H),7.16-7.12(m,1H),7.06-7.02(m,2H ),6.70(d,J=3.4Hz,1H),5.06-5.03(m,1H),4.58(dd,J=14.0,8.3Hz,1H),4.43-4.34(m,2H),4.22(d,J=12.0Hz,1H),4.03(s,2H).

[0096] Example 10: 3-(4-((1-(2,4-difluorophenyl)-2-(1H-1,2,4-triazol-1-yl)ethoxy)methyl)benzyl)-5-(4-trifluoromethylphenyl)isoxazole (E04)

[0097] Using intermediates Z-4d and Z-6a as raw materials, and following the synthesis method of intermediate E01, a yellow solid E04 was obtained. HRMS(ESI): Calcd.for C 28 H21 F5N4O2[M+Na] + :563.1477,Found 563.1492[M+Na] + . 1 H NMR (400MHz, DMSO-d6) δ8.45(s,1H),8.05(d,J=8.1Hz,2H),7.94(s,1H),7.86(d,J=8.3Hz,2H),7.52-7.46(m,1H),7.29-7.22(m,3H),7.17-7.1 2(m,1H),7.08-7.04(m,3H),5.06(dd,J=8.3,4.1Hz,1H),4.58(dd,J=14.1,8.3Hz,1H),4.44-4.35(m,2H),4.22(d,J=12.0Hz,1H),4.04(s,2H).

[0098] Example 11: 3-(4-((1-(2,4-difluorophenyl)-2-(1H-1,2,4-triazol-1-yl)ethoxy)methyl)benzyl)-5-(4-methylphenyl)isoxazole (E05)

[0099] Using intermediates Z-4e and Z-6a as raw materials, and following the synthesis method of intermediate E01, a yellow solid E05 was obtained. HRMS(ESI):Calcd.for C 28 H 24 F2N4O2[M+Na] + :509.1760,Found 509.1759[M+Na] + . 1 H NMR (400MHz, DMSO-d6) δ8.45 (s, 1H), 7.94 (s, 1H), 7.71 (d, J = 8.2Hz, 2H), 7. 49(td,J=8.5,6.6Hz,1H),7.33-7.20(m,5H),7.18-7.12(m,1H),7.05(d,J=8 .0Hz,2H),6.77(s,1H),5.05(dd,J=8.3,4.1Hz,1H),4.58(dd,J=14.3,8.3H z,1H),4.45-4.35(m,2H),4.22(d,J=11.9Hz,1H),3.99(s,2H),2.34(s,3H).

[0100] Example 12: 3-(4-((1-(2,4-difluorophenyl)-2-(1H-1,2,4-triazol-1-yl)ethoxy)methyl)benzyl)-5-(4-methoxyphenyl)isoxazole (E06)

[0101] Using intermediates Z-4f and Z-6a as raw materials, and following the synthesis method of intermediate E01, a yellow solid E06 was obtained. HRMS(ESI):Calcd.for C 28 H 24 F2N4O3[M+Na] + :525.1709,Found 525.1715[M+Na] + . 1 H NMR(400MHz,DMSO-d6)δ8.45(s,1H),7.94(s,1H),7.79-7.73(m,2H),7.49 (td,J=8.5,6.6Hz,1H),7.30-7.21(m,3H),7.17-7.12(m,1H),7.05(d,J=8. 8Hz,4H),6.69(s,1H),5.05(dd,J=8.3,4.2Hz,1H),4.58(dd,J=14.1,8.3Hz ,1H),4.45-4.35(m,2H),4.22(d,J=11.9Hz,1H),3.98(s,2H),3.81(s,3H).

[0102] Example 13: 3-(3-(4-(1-(2,4-difluorophenyl)-2-(1H-1,2,4-triazol-1-yl)ethoxy)methyl)benzyl)isoxazol-5-yl)pyridine-2-amine (E07)

[0103] Using 3-(3-bromomethyl)isoxazol-5-yl)pyridin-2-amine and intermediate Z-6a as starting materials, and following the synthetic method for intermediate E01, a yellow solid E07 was obtained. HRMS(ESI): Calcd.for C 26 H 22 F2N6O2[M+Na] + :511.1665,Found 511.1647[M+Na] + . 1H NMR(500MHz,Chloroform-d)δ8.31(dd,J=4.2,2.2Hz,1H),8.25(d,J=1.6Hz,1H),8.00(dd,J=9.2,2.2Hz,1H), 7.75(d,J=1.6Hz,1H),7.50(dtd,J=7.9,5.0,1.0Hz,1H),7.28-7.20(m,4H),7.09(dd,J=9.1,4.1Hz,1H),6.98 (dtd,J=19.6,8.0,1.9Hz,2H),6.68(d,J=8.2Hz,1H),6.50(d,J=8.2Hz,1H),6.23(s,1H),5.19(td,J=4.4,0.9 Hz,1H),4.70(q,J=0.9Hz,2H),4.45(dd,J=14.5,4.4Hz,1H),4.39(dd,J=14.5,4.4Hz,1H),4.16-4.08(m,2H).

[0104] Example 14: 3-(4-((1-(2,4-dichlorophenyl)-2-(1H-imidazol-1-yl)ethoxy)methyl)benzyl)-5-phenylisoxazole (E08)

[0105] Using intermediates Z-4a and Z-7b as raw materials, and following the synthesis method of intermediate E01, a yellow solid E08 was obtained. HRMS(ESI): Calcd.for C 28 H 23 Cl2N3O2[M+Na] + :526.1060,Found 526.1044[M+Na] + . 1 H NMR(500MHz,Chloroform-d)δ7.83-7.75(m,2H),7.70(t,J=1.7Hz,1H),7.54-7.36(m,6H),7.27-7.22(m,2H),7.25-7.19(m,3H) ,7.07(dd,J=4.1,1.7Hz,1H),6.74(s,1H),5.13(td,J=5.0,0.9Hz,1H),4.74-4.64(m,2H),4.45-4.34(m,2H),4.16-4.08(m,2H).

[0106] Example 15: 3-(4-((1-(2,4-dichlorophenyl)-2-(1H-imidazol)ethoxy)methyl)benzyl)-5-(4-fluorophenyl)isoxazole (E09)

[0107] Using intermediates Z-4b and Z-7b as raw materials, and referring to the synthesis method of intermediate E01, yellow solid E09 was obtained.

[0108] HRMS(ESI):Calcd.for C 28 H 22 Cl2FN3O2[M+Na] + :544.0965,Found544.0945[M+Na] + . 1 H NMR(500MHz,Chloroform-d)δ7.87-7.80(m,2H),7.70(t,J=1.7Hz,1H),7.52(d,J=2.0Hz,1H),7.41(qd,J=8.6,1.4Hz,2H),7.24(t,J=0.9Hz,2H), 7.25-7.13(m,5H),7.07(dd,J=4.1,1.7Hz,1H),6.74(s,1H),5.13(td,J= 5.0,0.9Hz,1H),4.74-4.64(m,2H),4.45-4.34(m,2H),4.16-4.08(m,2H).

[0109] Example 16: 3-(4-((1-(2,4-dichlorophenyl)-2-(1H-imidazol)ethoxy)methyl)benzyl)-5-(2,4-difluorophenyl)isoxazole (E10)

[0110] Using intermediates Z-4c and Z-7b as raw materials, and following the synthesis method of intermediate E01, a yellow solid E10 was obtained. HRMS(ESI): Calcd.for C 28 H 21 Cl2F2N3O2[M+Na] + :562.0871,Found 562.0864[M+Na] + . 1 H NMR(500MHz,Chloroform-d)δ7.78(dt,J=7.8,4.9Hz,1H),7.70(t,J=1.7Hz,1H),7.52(d,J=2.1Hz,1H),7.41(qd,J=8.6,1.4Hz,2H),7.28-7.22(m,2H ),7.25-7.19(m,3H),7.12-7.02(m,3H),6.79(d,J=2.0Hz,1H),5.13(td,J =5.0,0.9Hz,1H),4.74-4.64(m,2H),4.45-4.34(m,2H),4.16-4.08(m,2H).

[0111] Example 17: 3-(4-((1-(2,4-dichlorophenyl)-2-(1H-imidazol)ethoxy)methyl)benzyl)-5-(4-trifluoromethylphenyl)isoxazole (E11)

[0112] Using intermediates Z-4d and Z-7b as raw materials, and following the synthesis method of intermediate E01, a yellow solid E11 was obtained. HRMS(ESI): Calcd.for C 29 H 22 Cl2F3N3O2[M+Na] + :594.0933,Found 594.0920[M+Na] + . 1 H NMR(500MHz,Chloroform-d)δ7.91-7.84(m,2H),7.72-7.63(m,3H),7.52(d,J=2.0Hz,1H),7.41(qd,J=8.6,1.4Hz,2H),7.28-7.19(m ,5H),7.07(dd,J=4.1,1.7Hz,1H),6.78(s,1H),5.13(td,J=5.0,0.9Hz,1H),4.74-4.64(m,2H),4.45-4.34(m,2H),4.16-4.08(m,2H).

[0113] Example 18: 3-(4-((1-(2,4-dichlorophenyl)-2-(1H-imidazol)ethoxy)methyl)benzyl)-5-(4-methylphenyl)isoxazole (E12)

[0114] Using intermediates Z-4e and Z-7b as raw materials, and following the synthesis method of intermediate E01, a yellow solid E12 was obtained. HRMS(ESI):Calcd.for C 29 H 25 Cl2N3O2[M+Na] + :540.1216,Found 540.1210[M+Na] + . 1H NMR(500MHz,Chloroform-d)δ7.73-7.67(m,3H),7.52(d,J=2.0Hz,1H),7.45-7.36(m,2H),7.28-7.19(m,7H),7.07(dd,J=4.1,1.7 Hz,1H),6.73(s,1H),5.13(td,J=5.0,0.9Hz,1H),4.74-4.64(m,2H),4.45-4.34(m,2H),4.16-4.08(m,2H),2.40(d,J=1.5Hz,1H).

[0115] Example 19: 3-(4-((1-(2,4-dichlorophenyl)-2-(1H-imidazol)ethoxy)methyl)benzyl)-5-(4-methoxyphenyl)isoxazole (E13)

[0116] Using intermediates Z-4f and Z-7b as raw materials, and following the synthesis method of intermediate E01, a yellow solid E13 was obtained. HRMS(ESI):Calcd.for C 29 H 25 Cl2N3O3[M+Na] + :556.1165,Found 556.1147[M+Na] + . 1 H NMR(500MHz,Chloroform-d)δ7.80-7.75(m,2H),7.70(t,J=1.7Hz,1H),7.52(d,J=2.0Hz,1H),7.41(qd,J=8.6,1.4Hz,2H),7.28-7.19(m,4H),7.07(dd ,J=4.1,1.7Hz,1H),7.05-7.00(m,2H),6.72(s,1H),5.13(td,J=5.0,0.9Hz ,1H),4.74-4.64(m,2H),4.45-4.34(m,2H),4.16-4.08(m,2H),3.83(s,2H).

[0117] Example 20: 3-(3-(4-(1-(2,4-dichlorophenyl)-2-(1H-imidazol-1-yl)ethoxy)methyl)benzyl)isoxazol-5-yl)pyridine-2-amine (E14)

[0118] Using 3-(3-bromomethyl)isoxazol-5-yl)pyridin-2-amine and intermediate Z-6b as starting materials, and following the synthetic method for intermediate E01, yellow solid E14 was obtained. HRMS(ESI):Calcd.for C 27 H 23Cl2N5O2[M+Na] + :542.1121,Found542.1105[M+Na] + . 1 H NMR(500MHz,Chloroform-d)δ8.31(dd,J=4.2,2.2Hz,1H),8.00(dd,J=9.2,2.2Hz,1H),7.7 0(t,J=1.7Hz,1H),7.52(d,J=2.0Hz,1H),7.41(qd,J=8.6,1.4Hz,2H),7.28-7.21(m,4H),7 .24-7.19(m,1H),7.12-7.05(m,2H),6.68(d,J=8.2Hz,1H),6.50(d,J=8.2Hz,1H),6.23(s, 1H),5.13(td,J=5.0,0.9Hz,1H),4.74-4.64(m,2H),4.45-4.34(m,2H),4.16-4.08(m,2H).

[0119] Pharmacological studies of the present invention

[0120] The in vitro antifungal activity of compounds E01-E18 was determined using the two-fold dilution method.

[0121] The experiment was conducted in accordance with the 2003 edition of the antifungal susceptibility testing protocol for sporogenic filamentous fungi published by the National Committee for Clinical Laboratory Standards (NCCLS). RPMI-1640 was used for both culture and dilution.

[0122] Positive control drug: Fluconazole.

[0123] Test strains: All seven test fungi were commercially available and preserved and provided by the Department of Microbiology, Shenyang Pharmaceutical University. They were Candida albicans (CPCC400616 and ATCCSC5314), Cryptococcus neoformans (CGMCC2.3161), Candida zeylanoides (CGMCC2.3739), Candida krusei (AS2.1045), Candida glabrata (ATCC 22019), and Candida glabrata.

[0124] Preparation of stock solutions for target compounds and control drugs: Weigh 4 mg of the target compound (E01-E18) and the positive control drug, respectively, dissolve them in 5 mL of DMSO, add 5 mL of Tween 80 and 4.0 mL of distilled water to prepare stock solutions, and store them at 4 °C for later use.

[0125] Determination of minimum inhibitory concentration (MIC) in vitro: RPM1640 medium was added to a 96-cell plate. 180 μL was added to well 1, 100 μL to the growth control and experimental wells, and 200 μL to the blank control wells. Then, 20 μL of the drug stock solution was added to well 1, mixed well, and 100 μL was transferred to well 2. Another 100 μL was added to well 2 and transferred to well 3, and so on, until all experimental wells were reached. No drug stock solution was added to the growth control and blank control wells. Finally, 100 μL of the prepared drug stock solution, diluted to 1×10⁻⁶, was added to each of the experimental and growth control wells. 3 ~10 3 A bacterial suspension of cuf / mL was prepared. 96 plates were incubated at 35°C. MIC values ​​for *Candida albicans*, *Candida tropicalis*, *Candida krusei*, *Candida glabrata*, and *Candida glabrata* were measured after 24 hours of incubation. MIC values ​​for *Cryptococcus neoformans* were measured after 72 hours of incubation. All experiments were performed at least three times independently, with an experimental error not exceeding 10%.

[0126]

[0127] Note: MIC: minimum inhibitory concentration, Fluconazole, C. alb: Candida albicans, C. alb(5314): Candida albicans SC5314, C. par: Candida glabrata, C. glabrata: Candida glabrata, C. kru: Candida krubrata, C. tro: Candida tropicalis, C. neo: Cryptococcus neoformans.

[0128] As can be seen from the above, the E01-E14 compounds of this invention exhibit superior in vitro antifungal activity, with some compounds showing significantly better antifungal activity than positive control drugs, and also possessing a broader antibacterial spectrum. The azole derivatives containing isoxazole structures designed and synthesized in this invention possess strong antifungal activity, significantly improving their antibacterial effect compared to positive control drugs.

Claims

1. An azole derivative containing an isoxazole structure, characterized in that: The compound is shown in general formula (1). In the general formula: X, Y, and Z can be the same or different and are selected from N or CH; R 1 R 2 R 3 R 4 The same or different groups may be selected from hydrogen, halogen, cyano, hydroxyl, amino, unsubstituted or substituted by at least one of the following groups: C1-6 alkyl or C1-6 alkoxy, which may be the same or different groups selected from halogen, hydroxyl, or cyano.

2. The compound according to claim 1, characterized in that: In the general formula: X, Y, and Z can be the same or different and are selected from N or CH; R 1 R 2 R 3 R 4 The same or different groups may be selected from hydrogen atoms, halogens, cyano groups, hydroxyl groups, amino groups, unsubstituted or substituted C1-4 alkyl groups, and C1-4 alkoxy groups, which may be the same or different groups selected from halogens, hydroxyl groups, and cyano groups.

3. The compound according to claim 2, characterized in that: In the general formula: X, Y, and Z can be the same or different and are selected from N or CH; R 1 R 2 R 3 R 4 The same or different groups may be selected from hydrogen atoms, halogens, cyano groups, hydroxyl groups, amino groups, unsubstituted or substituted C1-4 alkyl groups, or C1-4 alkoxy groups, which may be the same or different groups selected from halogens or hydroxyl groups.

4. The compound according to claim 3, characterized in that: In the general formula: X, Y, and Z can be the same or different and are selected from N or CH; R 1 R 2 R 3 R 4 The same or different groups may be selected from hydrogen atoms, halogens, cyano groups, hydroxyl groups, amino groups, unsubstituted or substituted C1-4 alkyl groups, C1-4 alkoxy groups, which may be the same or different groups selected from halogens.

5. A method for preparing the compound of claim 1, wherein the reaction formula is as follows: firstly, acetophenone containing different substituents on the benzene ring reacts with diethyl oxalate to generate intermediate Z-1a-f, then reacts with hydroxylamine hydrochloride to generate isoxazole ring intermediate Z-2a-f, and then undergoes reduction and bromination to obtain intermediate Z-4a-f; compounds a and b respectively undergo substitution reactions with p-bromobenzyl bromide to generate intermediate Z-5a-b, and then undergo boronization and Suzuki reaction with intermediate Z-4a-f to obtain the compound of general formula (1); 。 6. A pharmaceutical composition, characterized in that: The composition comprises the compound of any one of claims 1-4 and a pharmaceutically acceptable carrier.

7. The use of the compound according to any one of claims 1-4 or the composition according to claim 6 in the preparation of an antifungal drug.

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