Azole derivative containing double-bond structure and application thereof

Abstract

The invention belongs to the technical field of medicines, and relates to an azole derivative containing a double-bond structure and medical application thereof. The compound is shown as a general formula (1), achieves strong and broad-spectrum antifungal activity, and has potential application in the aspect of antifungal drugs. R in the general formula (1) is defined in the specification. # imgabs0 #

Description

Technical Field

[0001] The invention belongs to the technical field of medicines and relates to an azole derivative containing a double bond structure and its medical application. Background Art

[0002] The current treatment of fungal infections faces many challenges. The morbidity and mortality of invasive fungal infections (IFIs) are increasing year by year, the resistance of antifungal drugs is becoming more serious, and the long course of antifungal treatment, as well as the safety and drug interactions of antifungal drugs, urgently need new antifungal drugs. Azoles have been widely used in clinical practice and have important value in fighting deep fungal infections. Azoles are cytochrome P450-14α-demethylase (CYP51) inhibitors. Their mechanism of action is to inhibit the biosynthesis of ergosterol on the fungal cell membrane, thereby leading to fungal cell death.

[0003] It is reported that the active site of CYP51 has three different parts: (a) azole ring (coordinated with the heme iron of CYP51); (b) hydrophobic channel I; (c) another narrow hydrophobic channel II (substrate entry channel). In the present invention, a series of azole derivatives containing double bond structures are designed, and the side chains of the compounds can more fully occupy the hydrophobic fragment II, so as to obtain CYP51 inhibitors with anti-resistance activity, more stable pharmacokinetic properties and low toxicity, and treat invasive fungal infections. Summary of the invention

[0004] The purpose of the present invention is to design and synthesize azole derivatives containing double bond structures, and their preparation methods and medical applications.

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

[0006] An azole derivative containing a double bond structure, characterized in that: the compound is as shown in the general formula (1)

[0007]

[0008] In the general formula:

[0009] X and Y may be the same or different and may be selected from N or CH;

[0010] R 1 , R 2 may be the same or different and be selected from H, halogen, cyano, nitro, hydroxyl,

[0011] C1-C6 alkyl, C3-C8 cycloalkyl, C2-C6 alkenyl, C2-C6 alkynyl, C1-C6 alkoxy, C2-C6 alkenyloxy, C2-C6 alkynyloxy, unsubstituted or substituted by at least one of the following groups, which may be the same or different and are selected from halogen, hydroxy, cyano, nitro or amino;

[0012] unsubstituted or substituted amino, such as the following groups, such as the following groups are selected from C1-C6 alkyl, C1-C6 aminoalkyl, C1-C6 hydroxyalkyl, C1-C6 alkoxyalkyl, C1-C6 cyanoalkyl, C2-C6 alkenyl, C2-C6 alkynyl, C1-C6 alkylsulfonyl, C1-C6 alkylcarbonyl, C2-C6 alkenylcarbonyl or C2-C6 alkynylcarbonyl;

[0013] An unsubstituted or substituted sulfamide or amide group, wherein the group replacing the amino group in the sulfamide or amide group is selected from C1-C6 alkyl, C1-C6 aminoalkyl, C1-C6 hydroxyalkyl, C1-C6 alkoxyalkyl, C1-C6 cyanoalkyl, C2-C6 alkenyl, C2-C6 alkynyl, C1-C6 alkylsulfonyl, C1-C6 alkylcarbonyl, C2-C6 alkenylcarbonyl or C2-C6 alkynylcarbonyl;

[0014] R 3 is selected from H, C1-C6 alkyl, C3-C8 cycloalkyl, C2-C6 alkenyl, C2-C6 alkynyl, C1-C6 alkoxy, C2-C6 alkenyloxy, C2-C6 alkynyloxy, which are unsubstituted or substituted by at least one of the following groups, which may be the same or different and are selected from halogen, hydroxyl, cyano, nitro or amino;

[0015] Phenyl, biphenyl, naphthyl, benzyl, pyridine ring, pyrazole ring, pyrrole ring, pyrimidine ring, quinoline ring, imidazole ring, morpholine ring, piperazine ring, pyridazine ring, pyrazine ring, piperidine ring, thiophene ring, pyran ring, indole ring, furan ring, which are unsubstituted or substituted with at least one of the following groups, the following groups may be the same or different and are selected from hydroxyl, hydroxymethyl, thiol, amino, sulfamide, carboxyl, ester, cyano, nitro, halogen, C1-C6 alkyl, C1-C6 alkoxy, C1-C6 alkylamino, C3-C7 cycloalkyl, benzyloxycarbonyl, C1-C6 haloalkyl;

[0016] Or, optical isomers or diastereomers of the compound represented by general formula (1).

[0017] Preferably, the compound has the general formula:

[0018] X and Y may be the same or different and may be selected from N or CH;

[0019] R 1 , R 2may be the same or different and be selected from H, halogen, cyano, nitro, hydroxyl,

[0020] C1-C6 alkyl, C3-C8 cycloalkyl, C2-C6 alkenyl, C2-C6 alkynyl, C1-C6 alkoxy, C2-C6 alkenyloxy, C2-C6 alkynyloxy, unsubstituted or substituted by at least one of the following groups, which may be the same or different and are selected from halogen, hydroxy, cyano, nitro and amino;

[0021] R 3 is selected from H, C1-C6 alkyl, C3-C8 cycloalkyl, C2-C6 alkenyl, C2-C6 alkynyl, C1-C6 alkoxy, C2-C6 alkenyloxy, C2-C6 alkynyloxy, which are unsubstituted or substituted by at least one of the following groups, which may be the same or different and are selected from halogen, hydroxyl, cyano, nitro or amino;

[0022] Phenyl, pyridine ring, pyrazole ring, pyrrole ring, pyrimidine ring, imidazole ring, piperidine ring, thiophene ring, pyran ring, indole ring, furan ring which are unsubstituted or substituted by at least one of the following groups, the following groups may be the same or different and are selected from cyano, nitro, halogen, C1-C6 alkyl, C1-C6 alkoxy, C1-C6 alkylamino, C3-C7 cycloalkyl, benzyloxycarbonyl, C1-C6 haloalkyl;

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

[0024] Further preferably, the compound has the general formula:

[0025] X and Y may be the same or different and may be selected from N or CH;

[0026] R 1 , R 2 which may be the same or different and are selected from H, halogen, cyano, nitro, hydroxyl, C1-C4 alkyl, C1-C4 alkoxy which are unsubstituted or substituted by at least one of the following groups, which may be the same or different and are selected from halogen, hydroxyl, cyano, nitro and amino;

[0027] R 3 Selected from H, C1-C6 alkyl, C3-C8 cycloalkyl which are unsubstituted or substituted by at least one of the following groups, which may be the same or different and are selected from halogen, nitro and amino; phenyl, pyrrole ring, thiophene ring, pyran ring, furan ring which are unsubstituted or substituted by at least one of the following groups, which may be the same or different and are selected from cyano, nitro, halogen, C1-C4 alkyl, C1-C4 alkoxy, C3-C7 cycloalkyl, C1-C6 haloalkyl;

[0028] Or, optical isomers or diastereomers of the compound represented by general formula (1).

[0029] Still further preferably, the compound has the general formula:

[0030] X and Y may be the same or different and may be selected from N or CH;

[0031] R 1 , R 2 which may be the same or different and are selected from hydrogen, halogen, cyano, nitro, hydroxyl, C1-4 alkyl, C1-4 alkoxy;

[0032] R 3 Selected from hydrogen atom; C1-C6 alkyl, C3-C8 cycloalkyl; phenyl, pyrrole ring, thiophene ring, pyran ring, furan ring which are unsubstituted or substituted by at least one of the following groups, the following groups may be the same or different and selected from halogen, C1-C4 alkyl, C1-C4 alkoxy, C1-C6 haloalkyl;

[0033] Or, optical isomers or diastereomers of the compound represented by general formula (1).

[0034] In further preference, the compound has the general formula:

[0035] X and Y may be the same or different and may be selected from N or CH;

[0036] R 1 , R 2 which may be the same or different and are selected from H, halogen, C1-C4 alkyl, C1-C4 alkoxy;

[0037] R 3 Selected from H, C3-C5 cycloalkyl; phenyl, pyrrole ring, thiophene ring, pyran ring, furan ring which are unsubstituted or substituted by at least one of the following groups, the following groups may be the same or different and selected from halogen, C1-C4 alkyl, C1-C4 alkoxy, trifluoromethyl;

[0038] Or, optical isomers or diastereomers of the compound represented by general formula (1).

[0039] Most preferably, the compound is the following D01-D20 compound and its optical isomers and diastereomers;

[0040]

[0041] A pharmaceutical composition comprises the compound represented by the general formula (1) and its optically active form, diastereoisomer and a pharmaceutically acceptable carrier.

[0042] A use of a compound and a composition, wherein the compound represented by the general formula (1) and its optically active form, diastereoisomer or the composition are used in the preparation of a drug for inhibiting CYP51.

[0043] The use of the compound represented by the general formula (1) and its optically active form, diastereoisomer or the composition in the preparation of antifungal drugs.

[0044] The advantages of the present invention are:

[0045] The double-bond-containing azole derivatives designed by the present invention have strong anti-resistance activity against fluconazole-sensitive bacteria and fluconazole-resistant bacteria. At the same time, the compound has good liver microsome stability and has potential application as an antifungal drug. DETAILED DESCRIPTION

[0046] The following examples will provide a better understanding of the compounds of the present invention and their preparation. These examples are intended to illustrate but not to limit the scope of the present invention.

[0047] Example 1: (E)-2-(4-((1-(2,4-difluorophenyl)-2-(1-imidazol-1-yl)ethoxy)methyl)phenylvinyl)-5-phenyl-1,3,4-oxadiazole (D01)

[0048] In a 50 mL eggplant-shaped bottle, 1-(2,4-difluorophenyl)-2-(1H-imidazole)ethanol (1a, 0.50 g, 2 mmol, 1 eq), 4-bromomethylcinnamic acid methyl ester (2, 0.57 g, 2 mmol, 1 eq), sodium hydride (0.13 g, 5 mmol, 2.5 eq), and tetrahydrofuran 5 mL were added. The mixture was heated at 70 °C for 3 h under stirring. 10 mL of water was added to the reaction solution, and the mixture was extracted with ethyl acetate (3 × 10 mL). The organic phases were combined, washed with water (3 × 10 mL) and saturated brine (10 mL) in turn, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to obtain a crude product, which was purified by column chromatography (dichloromethane: methanol = 20: 1) to obtain 3a as a yellow solid, with a yield of 0.78 g and a yield of 87.40%.

[0049] In a 50 mL eggplant-shaped bottle, add intermediate 3a (0.5 g, 1 mmol), 1N potassium hydroxide solution, and 5 mL of methanol. Stir and react at room temperature for 1 h. Distill off part of the solvent under reduced pressure, add 4N dilute hydrochloric acid to adjust the pH to 6, filter to obtain a solid, and recrystallize from ethanol to obtain 4a, which is a white solid with an amount of 0.41 g and a yield of 85.43%.

[0050] In a 50mL eggplant-shaped bottle, add intermediate 4a (0.5g, 1mmol, 1eq), benzoyl hydrazide (0.18g, 1mmol, 1eq), and 10mL of POCl3. Stir and react at 110°C for 6h. After the reaction solution is cooled to room temperature, 50mL of ice water is added to the reaction solution, and 1N potassium hydroxide solution is added to adjust the pH to 7. Extract with dichloromethane (3×30mL), combine the organic phases, wash with water (3×20mL) and saturated brine (20mL) in turn, dry with anhydrous sodium sulfate, filter the desiccant, and concentrate under reduced pressure to obtain a crude product, which is purified by column chromatography (dichloromethane: methanol = 20:1) to obtain D01 as a yellow solid, with an amount of 0.21g and a yield of 33.40%. 1 H NMR(400MHz, DMSO-d6)δ8.10(dd,J=7.5,2.1Hz,2H),7.81-7.68(m,3H),7.63(dd,J=6.3,1.3Hz,3H),7.53(s,1H),7.45-7.34(m,2H),7.30-7.22 (m,3H),7.15-7.06(m,2H),6.88(d,J=1.1Hz,1H),4.96(dd,J=7.2,4.6Hz,1H),4.44(d,J=12.5Hz,1H),4.41-4.29(m,3H).HRMS(ESI):Calcd.for C 28 H 22 F2N4O2[M+Na] + :507.1603,Found 507.1571[M+Na] + .

[0051] Example 2: (E)-2-(4-((1-(2,4-difluorophenyl)-2-(1-imidazol-1-yl)ethoxy)methyl)phenylvinyl)-5-(4-methylphenyl)-1,3,4-oxadiazole (D02)

[0052] The preparation method of the compound of Example 2 is the same as that of Example 1, using intermediate 4a and p-methylbenzoyl hydrazide as raw materials, and referring to the synthesis method of D01, D02 is obtained as a yellow solid. 1H NMR (400MHz, DMSO-d6) δ7.99(d,J=8.3Hz,2H),7.77-7.69(m,3H),7.53(s,1H),7.46-7.23(m,7H),7.17-7.07(m,2H),6.86(d ,J=10.7Hz,1H),4.96(dd,J=7.1,4.6Hz,1H),4.45(d,J=12.5Hz,1H),4.42-4.28(m,3H),2.41(s,3H).HRMS(ESI):Calcd.for C 29 H 24 F2N4O2[M+Na] + :521.1760,Found

[0053] 521.1769[M+Na] + .

[0054] Example 3: (E)-2-(4-((1-(2,4-difluorophenyl)-2-(1-imidazol-1-yl)ethoxy)methyl)phenylvinyl)-5-(4-methoxyphenyl)-1,3,4-oxadiazole (D03)

[0055] The preparation method of the compound of Example 3 is the same as that of Example 1, using intermediate 4a and p-methoxybenzoic acid hydrazide as raw materials, and referring to the synthesis method of D01, D03 is obtained as a yellow solid. 1 H NMR(400MHz, DMSO-d6)δ8.03(d,J=8.8Hz,2H),7.73-7.66(m,3H),7.54(s,1H),7.45-7.39(m,1H),7.33-7.0 9(m,8H),6.88(s,1H),4.96(dd,J=7.1,4.5Hz,1H),4.46-4.30(m,4H),3.86(s,3H).HRMS(ESI):Calcd.forC 29 H 24 F2N4O3[M+Na] + :537.1709,Found 537.1713[M+Na] + .

[0056] Example 4: (E)-2-(4-((1-(2,4-difluorophenyl)-2-(1-imidazol-1-yl)ethoxy)methyl)phenylvinyl)-5-(4-fluorophenyl)-1,3,4-oxadiazole (D04)

[0057] The preparation method of the compound of Example 4 is the same as that of Example 1, using intermediate 4a and p-fluorobenzoyl hydrazide as raw materials, and referring to the synthesis method of D01, D04 is obtained as a yellow solid.1 H NMR (400MHz, DMSO-d6) δ8.15(dd,J=8.9,5.3Hz,2H),7.77-7.70(m,3H),7.54(s,1H),7.47(t,J=8.8Hz,2H),7.43-7.38(m,1H),7.36(d,J=16.5Hz,1H ),7.30-7.22(m,3H),7.15-7.08(m,2H),6.88(s,1H),4.96(dd,J=7.1,4.5Hz,1H),4.44(d,J=12.5Hz,1H),4.39-4.28(m,3H).HRMS(ESI):Calcd.for C 28 H 21 F3N4O2[M+Na] + :525.1509,Found 525.1499[M+Na] + .

[0058] Example 5: (E)-2-(4-((1-(2,4-difluorophenyl)-2-(1-imidazol-1-yl)ethoxy)methyl)phenylvinyl)-5-(4-chlorophenyl)-1,3,4-oxadiazole (D05)

[0059] The preparation method of the compound of Example 5 is the same as that of Example 1, using intermediate 4a and p-chlorobenzoyl hydrazide as raw materials, and referring to the synthesis method of D01, D05 is obtained as a yellow solid. 1 H NMR(400MHz,DMSO-d6)δ8.15-8.03(m,2H),7.77-7.67(m,5H),7.53(s,1H),7.44-7.33(m,2H),7.29-7.22(m,3H), 7.11(dt,J=13.7,1.1Hz,2H),6.87(s,1H),4.96(dd,J=7.2,4.6Hz,1H),4.46-4.30(m,4H).HRMS(ESI):Calcd.forC 28 H 21 ClF2N4O2[M+Na] + :541.1213,Found 541.1194[M+Na] + .

[0060] Example 6: (E)-2-(4-((1-(2,4-difluorophenyl)-2-(1-imidazol-1-yl)ethoxy)methyl)phenylvinyl)-5-(4-bromophenyl)-1,3,4-oxadiazole (D06)

[0061] The preparation method of the compound of Example 6 is the same as that of Example 1, using intermediate 4a and p-bromobenzoylhydrazide as raw materials, and referring to the synthesis method of D01, D06 is obtained as a yellow solid. 1 H NMR (400MHz, DMSO-d6) δ8.02(d,J=8.6Hz,2H),7.83(d,J=8.6Hz,2H),7.76-7.69(m,3H),7.53(s,1H),7.46-7.39(m,1H),7.39-7.33(m,1H),7. 30-7.22(m,3H),7.16-7.08(m,2H),6.87(s,1H),4.96(dd,J=7.1,4.6Hz,1H),4.44(d,J=12.5Hz,1H),4.39-4.28(m,3H).HRMS(ESI):Calcd.for C 28 H 21 BrF2N4O2[M+Na] + :585.0708,Found 585.0713[M+Na] + .

[0062] Example 7: (E)-2-(4-((1-(2,4-difluorophenyl)-2-(1-imidazol-1-yl)ethoxy)methyl)phenylvinyl)-5-(4-trifluoromethylphenyl)-1,3,4-oxadiazole (D07)

[0063] The preparation method of the compound of Example 7 is the same as that of Example 1, using intermediate 4a and p-trifluoromethylbenzoylhydrazide as raw materials, and referring to the synthesis method of D01, D07 is obtained as a yellow solid. 1 H NMR (400MHz, DMSO-d6) δ8.31(d,J=8.1Hz,2H),8.01(d,J=8.3Hz,2H),7.83-7.73(m,3H),7.55(s,1H),7.42(dd,J=15.8,8.3Hz,2H),7.31-7 .24(m,3H),7.17-7.10(m,2H),6.89(s,1H),4.98(dd,J=7.2,4.6Hz,1H),4.46(d,J=12.6Hz,1H),4.41-4.30(m,3H).HRMS(ESI):Calcd.for C 29 H 21 F5N4O2[M+Na] + :575.1477,Found 575.1476[M+Na] + .

[0064] Example 8: (E)-2-(4-((1-(2,4-difluorophenyl)-2-(1-imidazol-1-yl)ethoxy)methyl)phenylvinyl)-5-(furan-2-yl)-1,3,4-oxadiazole (D08)

[0065] The preparation method of the compound of Example 8 is the same as that of Example 1, using intermediate 4a and 2-furoylcarboxylic acid hydrazide as raw materials, and referring to the synthesis method of D01, D08 is obtained as a yellow solid. 1 H NMR (400MHz, DMSO-d6) δ8.08 (dd, J=1.8, 0.8Hz, 1H), 7.72 (d, J=8.2Hz, 2H), 7.6 4(d,J=16.4Hz,1H),7.53(d,J=1.1Hz,1H),7.45-7.33(m,3H),7.29-7.21(m,3H ),7.15-7.08(m,2H),6.87(s,1H),6.82(dd,J=3.6,1.8Hz,1H),4.96(dd,J=7.1 ,4.5Hz,1H),4.44(d,J=12.5Hz,1H),4.39-4.28(m,3H).HRMS(ESI):Calcd.forC 26 H 20 F2N4O3[M+Na] + :497.1396,Found 497.1393[M+Na] + .

[0066] Example 9: (E)-2-(4-((1-(2,4-difluorophenyl)-2-(1-imidazol-1-yl)ethoxy)methyl)phenylvinyl)-5-(thiophen-2-yl)-1,3,4-oxadiazole (D09)

[0067] The preparation method of the compound of Example 9 is the same as that of Example 1, using intermediate 4a and 2-thiophenecarboxylic acid hydrazide as raw materials, referring to the synthesis method of D01, to obtain D09 as a yellow solid. 1 H NMR (400MHz, DMSO-d6) δ7.96(dd,J=5.0,1.2Hz,1H),7.90(dd,J=3.7,1.2Hz,1H),7.72(d,J=8.0Hz,2H),7.67(d,J=16.5Hz,1H),7.53(s,1H),7.46-7 .22(m,6H),7.15-7.08(m,2H),6.88(d,J=1.2Hz,1H),4.96(dd,J=7.1,4.6Hz,1H),4.44(d,J=12.5Hz,1H),4.39-4.29(m,3H).HRMS(ESI):Calcd.for C26 H 20 F2N4O2S[M+Na] + :513.1167,Found 513.1157[M+Na] + .

[0068] Example 10: (E)-2-Cyclopropyl-5-(4-((1-(2,4-difluorophenyl)-2-(1H-imidazol-1-yl)ethoxy)methyl)phenylvinyl)-1,3,4-oxadiazole (D10)

[0069] The preparation method of the compound of Example 10 is the same as that of Example 1, using intermediate 4a and cyclopropylcarboxyhydrazide as raw materials, referring to the synthesis method of D01, to obtain D10 as a yellow solid.

[0070] Example 11: (E)-2-(4-((1-(2,4-dichlorophenyl)-2-(1-imidazol-1-yl)ethoxy)methyl)phenylvinyl)-5-phenyl-1,3,4-oxadiazole (D01)

[0071] The preparation method of the compound of Example 11 is the same as that of Example 1, using intermediate 4b and benzoyl hydrazide as raw materials, and referring to the synthesis method of D01, D11 is obtained as a yellow solid. 1 H NMR(400MHz, DMSO-d6)δ9.14(s,1H),8.09(d,J=5.9Hz,2H),7.74-7.25(m,14H),5.17(d,J=6.0Hz,1H),4.61-4.37(m,4H).HRMS(ESI):Calcd.forC 28 H 22 Cl2N4O2[M+H] + :517.1193,Found 517.1178[M+H] + .

[0072] Example 12: (E)-2-(4-((1-(2,4-dichlorophenyl)-2-(1-imidazol-1-yl)ethoxy)methyl)phenylvinyl)-5-(4-methylphenyl)-1,3,4-oxadiazole (D12)

[0073] The preparation method of the compound of Example 12 is the same as that of Example 1, using intermediate 4b and p-methylbenzoyl hydrazide as raw materials, and referring to the synthesis method of D01, D12 is obtained as a yellow solid. 1H NMR (400MHz, DMSO-d6) δ7.99 (d, J=8.2Hz, 2H), 7.75-7.71 (m, 3H), 7.67 (d, J= 2.1Hz,1H),7.54(s,1H),7.49(dd,J=8.4,2.1Hz,1H),7.45-7.34(m,4H),7.2 5(d,J=8.0Hz,2H),7.07(s,1H),6.89(s,1H),5.03(dd,J=6.9,4.0Hz,1H),4. 46(d,J=12.5Hz,1H),4.38-4.27(m,3H),2.42(s,3H).HRMS(ESI):Calcd.forC 29 H 24 Cl2N4O2[M+H] + :531.1349,Found 531.1285[M+H] + .

[0074] Example 13: (E)-2-(4-((1-(2,4-dichlorophenyl)-2-(1-imidazol-1-yl)ethoxy)methyl)phenylvinyl)-5-(4-methoxyphenyl)-1,3,4-oxadiazole (D13)

[0075] The preparation method of the compound of Example 13 is the same as that of Example 1, using intermediate 4b and p-methoxybenzoic acid hydrazide as raw materials, and referring to the synthesis method of D01, D13 is obtained as a yellow solid. 1 H NMR(400MHz,DMSO-d6)δ8.04(d,J=8.9Hz,2H),7.74-7.68(m,3H),7.67(d,J=2.1Hz,1H), 7.54(s,1H),7.49(dd,J=8.4,2.1Hz,1H),7.42(d,J=8.4Hz,1H),7.34(d,J=16.4Hz,1H),7 .25(d,J=8.0Hz,2H),7.17(d,J=8.9Hz,2H),7.07(s,1H),6.89(s,1H),5.03(dd,J=6.9,4 .0Hz,1H),4.46(d,J=12.5Hz,1H),4.38-4.27(m,3H),3.87(s,3H).HRMS(ESI):Calcd.for C 29 H 24 Cl2N4O3[M+

[0076] Na] + :569.1118,Found 569.1117[M+Na] + .

[0077] Example 14: (E)-2-(4-((1-(2,4-dichlorophenyl)-2-(1-imidazol-1-yl)ethoxy)methyl)phenylvinyl)-5-(4-fluorophenyl)-1,3,4-oxadiazole (D14)

[0078] The preparation method of the compound of Example 14 is the same as that of Example 1, using intermediate 4b and p-fluorobenzoyl hydrazide as raw materials, and referring to the synthesis method of D01, D14 is obtained as a yellow solid. 1 H NMR(400MHz, DMSO-d6)δ8.16(dd,J=8.9,5.3Hz,2H),7.77-7.72(m,3H),7.67(d,J=2.1Hz,1H),7.55(s,1H),7.51-7.46(m,3H),7.43-7.34(m,2H) ,7.26(d,J=8.0Hz,2H),7.07(s,1H),6.90(s,1H),5.04(dd,J=6.9,4.0Hz,1H),4.46(d,J=12.5Hz,1H),4.38-4.28(m,3H).HRMS(ESI):Calcd.for C 28 H 21 Cl2FN4O2[M+Na] + :557.0918,Found 557.0876[M+Na] + .

[0079] Example 15: (E)-2-(4-((1-(2,4-dichlorophenyl)-2-(1-imidazol-1-yl)ethoxy)methyl)phenylvinyl)-5-(4-chlorophenyl)-1,3,4-oxadiazole (D15)

[0080] The preparation method of the compound of Example 15 is the same as that of Example 1, using intermediate 4b and p-chlorobenzoyl hydrazide as raw materials, and referring to the synthesis method of D01, D15 is obtained as a yellow solid. 1 H NMR (400MHz, DMSO-d6) δ8.11(d,J=8.6Hz,2H),7.77-7.66(m,7H),7.54(s,1H),7.48(dd,J=8.4,2.1Hz,1H),7.43-7.35(m,2H),7.25(d,J=8.0Hz,2H ),7.07(s,1H),6.89(s,1H),5.03(dd,J=6.9,4.0Hz,1H),4.46(d,J=12.5Hz,1H),4.36(d,J=12.4Hz,1H),4.33-4.26(m,2H).HRMS(ESI):Calcd.for C 28 H21 Cl3N4O2[M+H] + :551.0803,Found 551.0746[M+H] + .

[0081] Example 16: (E)-2-(4-((1-(2,4-dichlorophenyl)-2-(1-imidazol-1-yl)ethoxy)methyl)phenylvinyl)-5-(4-bromophenyl)-1,3,4-oxadiazole (D16)

[0082] The preparation method of the compound of Example 16 is the same as that of Example 1, using intermediate 4b and p-bromobenzoylhydrazide as raw materials, and referring to the synthesis method of D01, D16 is obtained as a yellow solid. 1 H NMR (400MHz, DMSO-d6) δ8.11(d,J=8.6Hz,2H),7.74-7.66(m,6H),7.54(s,1H),7.48(dd,J=8.4,2.1Hz,1H),7.43-7.35(m,2H),7.25(d,J=8.0Hz,2H ),7.07(s,1H),6.89(s,1H),5.03(dd,J=6.9,4.0Hz,1H),4.46(d,J=12.5Hz,1H),4.36(d,J=12.4Hz,1H),4.33-4.26(m,2H).HRMS(ESI):Calcd.for C 28 H 21 BrCl2N4O2[M+H] + :595.0298,Found 595.0232[M+H] + .

[0083] Example 17: (E)-2-(4-((1-(2,4-dichlorophenyl)-2-(1-imidazol-1-yl)ethoxy)methyl)phenylvinyl)-5-(4-trifluoromethylphenyl)-1,3,4-oxadiazole (D17)

[0084] The preparation method of the compound of Example 17 is the same as that of Example 1, using intermediate 4b and trifluoromethylbenzoyl hydrazide as raw materials, and referring to the synthesis method of D01, D17 is obtained as a yellow solid. 1H NMR(400MHz,DMSO-d6)δ8.30(d,J=8.1Hz,2H),7.99(d,J=8.2Hz,2H),7.82-7.7 2(m,3H),7.66(d,J=2.1Hz,1H),7.57(s,1H),7.48(dd,J=8.4,2.1Hz,1H),7.43 -7.37(m,2H),7.26(d,J=8.0Hz,2H),7.08(s,1H),6.91(s,1H),5.04(dd,J=6.9 ,3.9Hz,1H),4.46(d,J=12.6Hz,1H),4.38-4.28(m,3H).HRMS(ESI):Calcd.for C 29 H 21 Cl2F3N4O2[M+Na] + :607.0886,Found 607.0837[M+Na] + .

[0085] Example 18: (E)-2-(4-((1-(2,4-dichlorophenyl)-2-(1-imidazol-1-yl)ethoxy)methyl)phenylvinyl)-5-(furan-2-yl)-1,3,4-oxadiazole (D18)

[0086] The preparation method of the compound of Example 18 is the same as that of Example 1, using intermediate 4b and 2-furoylhydrazide as raw materials, referring to the synthesis method of D01, to obtain D18 as a yellow solid. 1 H NMR(400MHz, DMSO-d6)δ8.08(d,J=1.7Hz,1H),7.72(d,J=8.0Hz,2H),7.67-7.62(m ,2H),7.53(d,J=1.1Hz,1H),7.47(dd,J=8.4,2.1Hz,1H),7.42-7.34(m,3H),7.24( d,J=8.0Hz,2H),7.07(s,1H),6.89(s,1H),6.82(dd,J=3.6,1.8Hz,1H),5.02(dd,J =6.9,3.9Hz,1H),4.45(d,J=12.5Hz,1H),4.36-4.23(m,3H).HRMS(ESI):Calcd.for C 26 H 20 Cl2N4O3[M+H] + :507.0985,Found 507.0938[M+H] + .

[0087] Example 19: (E)-2-(4-((1-(2,4-dichlorophenyl)-2-(1-imidazol-1-yl)ethoxy)methyl)phenylvinyl)-5-(thiophen-2-yl)-1,3,4-oxadiazole (D19)

[0088] The preparation method of the compound of Example 19 is the same as that of Example 1, using intermediate 4b and 2-thiophenecarboxylic acid hydrazide as raw materials, referring to the synthesis method of D01, to obtain D19 as a yellow solid. 1 H NMR (400MHz, DMSO-d6) δ7.96 (dd, J=5.0, 1.2Hz, 1H), 7.90 (dd, J=3.7, 1.2Hz, 1H ),7.74-7.64(m,5H),7.48(dd,J=8.4,2.1Hz,1H),7.41(d,J=8.4Hz,1H),7.37- 7.31(m,2H),7.24(d,J=8.0Hz,2H),7.11(s,1H),6.92(s,1H),5.03(dd,J=6.8, 4.1Hz,1H),4.46(d,J=12.5Hz,1H),4.36-4.27(m,3H).HRMS(ESI):Calcd.forC 26 H 20 Cl2N4O2S[M+H] + :523.0757,Found 523.0714[M+H] + .

[0089] Example 20: (E)-2-Cyclopropyl-5-(4-((1-(2,4-dichlorophenyl)-2-(1H-imidazol-1-yl)ethoxy)methyl)phenylvinyl)-1,3,4-oxadiazole (D20)

[0090] The preparation method of the compound of Example 20 is the same as that of Example 1, using intermediate 4b and cyclopropylcarboxylic acid hydrazide as raw materials, and referring to the synthesis method of D01, D20 is obtained as a yellow solid. 1H NMR (400MHz, DMSO-d6) δ7.73-7.64(m,3H),7.56-7.46(m,3H),7.40(d,J=8.4Hz ,1H),7.31-7.20(m,3H),7.06(s,1H),6.88(s,1H),5.02(ddd,J=5.5,4.0,1.4H z,1H),4.44(d,J=12.4Hz,1H),4.36-4.23(m,3H),3.78(t,J=6.4Hz,1H),3.04( t,J=7.4Hz,1H),2.24-2.19(m,1H),1.23-1.05(m,2H).HRMS(ESI):Calcd.forC 25 H 22 Cl2N4O2[M+H] + :481.1193,Found 481.1134[M+H] + .

[0091] Pharmacological studies of the present invention

[0092] The in vitro antifungal activities of compounds D01-D20 were determined by the two-fold dilution method.

[0093] The experiment followed the 2003 edition of the spore-forming filamentous fungi susceptibility test protocol published by the National Committee for Clinical Laboratory Standards (NCCLS) of the United States. SDA medium was used as the culture medium for culture and dilution instead of RPMI-1640.

[0094] Positive control drugs: Fluconazole and Miconazole.

[0095] Test strains: All 7 test fungi can be purchased through relevant channels and then preserved by the Microbiology Teaching and Research Department of Shenyang Pharmaceutical University, including Candida albicans (CPCC400616 and ATCCSC5314), Cryptococcus neoformans (CGMCC2.3161), Candida zeylanoides CGMCC2.3739, Candida krusei (AS2.1045), Candida glabrata (ATCC 22019) and Candida glabrata. 5 fluconazole-resistant strains, including fluconazole-resistant strain 17#, fluconazole-resistant strain CaR, fluconazole-resistant strain 901, fluconazole-resistant strain 904 and fluconazole-resistant strain 632.

[0096] Preparation of target compound and control drug stock solutions: Weigh 4 mg of target compound and positive control drug respectively, dissolve in 5 mL of DMSO, add 5 mL of Tween 80 and 4.0 mL of distilled water to prepare stock solutions, and store at 4°C for future use.

[0097] Determination of in vitro minimum inhibitory concentration: Add RPM1640 culture medium to the 96-well plate, add 180 μL to the first well, add 100 μL to the growth control well and the experimental well, and add 200 μL to the blank control well. Then add 20 μL of the drug sample stock solution to the first well, mix well, and then aspirate 100 μL, and transfer it to the second well. After mixing well in the second well, aspirate 100 μL again, transfer it to the third well, and so on to all the experimental wells. No drug sample stock solution is added to the growth control well and the blank control well. Then add 100 μL of the prepared and diluted to 1×10 3 ~5×10 3 cuf / mL bacterial suspension. The 96-plate was cultured in a 35°C incubator. The MIC values ​​of Candida albicans, Candida tropicalis, Candida krusei, Candida glabrata, Candida glabrata and fluconazole-resistant bacteria were read after 24 hours of culture, and the MIC values ​​of Cryptococcus neoformans were read after 72 hours of culture. All experiments were performed at least three times independently, and the experimental error did not exceed 10%.

[0098]

[0099]

[0100]

[0101] Note: MIC: minimum inhibitory concentration, Fluconazole: fluconazole, Miconazole: miconazole, C.alb: Candida albicans, C.alb(5314): Candida albicans SC5314, C.par: Candida glabrata, C.gla: Candida glabrata, C.kru: Candida krusei, C.tro: Candida tropicalis, C.neo: Cryptococcus neoformans.

[0102] It can be seen from the above that D01-D20 of the present invention has strong in vitro antifungal activity against fluconazole-sensitive bacteria and fluconazole-resistant bacteria, and the antifungal activity is better than that of positive drugs and shows a wider antibacterial spectrum. The azole derivatives containing double bond structures designed and synthesized by the present invention have strong antifungal activity and significantly improve their antibacterial effect compared with positive drugs.

[0103] Study on the stability of liver microsomes of the present invention

[0104] Take the liver microsomes out of the -80℃ freezer, preheat and incubate them on a 37℃ water bath thermostat for 3 minutes, and melt them for use. Weigh a certain amount of NADPH, add an appropriate amount of magnesium chloride solution, and dissolve it into a 2mM solution for use. Prepare the incubation system mixture (without β-NADPH) and distribute it at a volume of 40μL / tube. 0min sample: add 240μL internal standard working precipitate, then add 40μL NADPH solution (negative control adds 40μL MgCl2 solution). Other samples: add 40μL NADPH solution to start the reaction (negative control group adds 40μL MgCl2 solution), then add 240μL internal standard precipitate, and incubate in a 37℃ water bath for 5, 15, 30 and 60 minutes. For the positive control group, add 40μL NADPH solution to start the reaction, incubate in a 37℃ water bath for 5, 15 and 60 minutes, and then add 240μL internal standard precipitant. All samples are vortexed and centrifuged. 150 μL of supernatant was added to 150 μL of water, vortexed to mix, and then the sample was injected into LC-MS / MS for analysis.

[0105]

[0106] It can be seen from the above that the metabolic rate of the representative compound D17 of the present invention is slower than that of miconazole, and it has better in vitro metabolic stability than miconazole. The in vitro metabolic stability of the azole derivatives containing double bond structures designed and synthesized by the present invention is better than that of positive drugs.

Claims

1. An azole derivative containing a double bond structure, characterized in that: The compound is shown in the general formula (1): In the general formula: X and Y may be the same or different and may be selected from N or CH; R 1 , R 2 may be the same or different and be selected from H, halogen, cyano, nitro, hydroxyl, C1-C6 alkyl, C3-C8 cycloalkyl, C2-C6 alkenyl, C2-C6 alkynyl, C1-C6 alkoxy, C2-C6 alkenyloxy, C2-C6 alkynyloxy, unsubstituted or substituted by at least one of the following groups, which may be the same or different and are selected from halogen, hydroxy, cyano, nitro or amino; unsubstituted or substituted amino, such as the following groups, such as the following groups are selected from C1-C6 alkyl, C1-C6 aminoalkyl, C1-C6 hydroxyalkyl, C1-C6 alkoxyalkyl, C1-C6 cyanoalkyl, C2-C6 alkenyl, C2-C6 alkynyl, C1-C6 alkylsulfonyl, C1-C6 alkylcarbonyl, C2-C6 alkenylcarbonyl or C2-C6 alkynylcarbonyl; An unsubstituted or substituted sulfamide or amide group, wherein the group replacing the amino group in the sulfamide or amide group is selected from C1-C6 alkyl, C1-C6 aminoalkyl, C1-C6 hydroxyalkyl, C1-C6 alkoxyalkyl, C1-C6 cyanoalkyl, C2-C6 alkenyl, C2-C6 alkynyl, C1-C6 alkylsulfonyl, C1-C6 alkylcarbonyl, C2-C6 alkenylcarbonyl or C2-C6 alkynylcarbonyl; R 3 is selected from H, C1-C6 alkyl, C3-C8 cycloalkyl, C2-C6 alkenyl, C2-C6 alkynyl, C1-C6 alkoxy, C2-C6 alkenyloxy, C2-C6 alkynyloxy, which are unsubstituted or substituted by at least one of the following groups, which may be the same or different and are selected from halogen, hydroxyl, cyano, nitro or amino; Phenyl, biphenyl, naphthyl, benzyl, pyridine ring, pyrazole ring, pyrrole ring, pyrimidine ring, quinoline ring, imidazole ring, morpholine ring, piperazine ring, pyridazine ring, pyrazine ring, piperidine ring, thiophene ring, pyran ring, indole ring, furan ring, which are unsubstituted or substituted with at least one of the following groups, the following groups may be the same or different and are selected from hydroxyl, hydroxymethyl, thiol, amino, sulfamide, carboxyl, ester, cyano, nitro, halogen, C1-C6 alkyl, C1-C6 alkoxy, C1-C6 alkylamino, C3-C7 cycloalkyl, benzyloxycarbonyl, C1-C6 haloalkyl; Or, optical isomers or diastereomers of the compound represented by general formula (1).

2. The compound according to claim 1, characterized in that: In the general formula: X and Y may be the same or different and may be selected from N or CH; R 1 , R 2 may be the same or different and be selected from H, halogen, cyano, nitro, hydroxyl, C1-C6 alkyl, C3-C8 cycloalkyl, C2-C6 alkenyl, C2-C6 alkynyl, C1-C6 alkoxy, C2-C6 alkenyloxy, C2-C6 alkynyloxy, unsubstituted or substituted by at least one of the following groups, which may be the same or different and are selected from halogen, hydroxy, cyano, nitro and amino; R 3 is selected from H, C1-C6 alkyl, C3-C8 cycloalkyl, C2-C6 alkenyl, C2-C6 alkynyl, C1-C6 alkoxy, C2-C6 alkenyloxy, C2-C6 alkynyloxy, which are unsubstituted or substituted by at least one of the following groups, which may be the same or different and are selected from halogen, hydroxyl, cyano, nitro or amino; Phenyl, pyridine ring, pyrazole ring, pyrrole ring, pyrimidine ring, imidazole ring, piperidine ring, thiophene ring, pyran ring, indole ring, furan ring which are unsubstituted or substituted by at least one of the following groups, the following groups may be the same or different and are selected from cyano, nitro, halogen, C1-C6 alkyl, C1-C6 alkoxy, C1-C6 alkylamino, C3-C7 cycloalkyl, benzyloxycarbonyl, C1-C6 haloalkyl; Or, optical isomers or diastereomers of the compound represented by general formula (1).

3. The compound according to claim 2, characterized in that: In the general formula: X and Y may be the same or different and may be selected from N or CH; R 1 , R 2 which may be the same or different and are selected from H, halogen, cyano, nitro, hydroxyl, C1-C4 alkyl, C1-C4 alkoxy which are unsubstituted or substituted by at least one of the following groups, which may be the same or different and are selected from halogen, hydroxyl, cyano, nitro and amino; R 3 Selected from H, C1-C6 alkyl, C3-C8 cycloalkyl which are unsubstituted or substituted by at least one of the following groups, which may be the same or different and are selected from halogen, nitro and amino; phenyl, pyrrole ring, thiophene ring, pyran ring, furan ring which are unsubstituted or substituted by at least one of the following groups, which may be the same or different and are selected from cyano, nitro, halogen, C1-C4 alkyl, C1-C4 alkoxy, C3-C7 cycloalkyl, C1-C6 haloalkyl; Or, optical isomers or diastereomers of the compound represented by general formula (1).

4. The compound according to claim 3, characterized in that: In the general formula: X and Y may be the same or different and may be selected from N or CH; R 1 , R 2 which may be the same or different and are selected from hydrogen, halogen, cyano, nitro, hydroxyl, C1-4 alkyl, C1-4 alkoxy; R 3 Selected from hydrogen atom; C1-C6 alkyl, C3-C8 cycloalkyl; phenyl, pyrrole ring, thiophene ring, pyran ring, furan ring which are unsubstituted or substituted by at least one of the following groups, the following groups may be the same or different and selected from halogen, C1-C4 alkyl, C1-C4 alkoxy, C1-C6 haloalkyl; Or, optical isomers or diastereomers of the compound represented by general formula (1).

5. The compound according to claim 4, characterized in that: In the general formula: X and Y may be the same or different and may be selected from N or CH; R 1 , R 2 which may be the same or different and are selected from H, halogen, C1-C4 alkyl, C1-C4 alkoxy; R 3 Selected from H, C3-C5 cycloalkyl; phenyl, pyrrole ring, thiophene ring, pyran ring, furan ring which are unsubstituted or substituted by at least one of the following groups, the following groups may be the same or different and selected from halogen, C1-C4 alkyl, C1-C4 alkoxy, trifluoromethyl; Or, optical isomers or diastereomers of the compound represented by general formula (1).

6. The compound according to claim 5, characterized in that: The compound represented by the general formula (1) is the following D01-D20 compounds and their optical isomers and diastereomers; 7. A pharmaceutical composition, characterized in that: The composition comprises the compound of general formula (1) as claimed in any one of claims 1 to 6 and its optically active form, diastereomer and a pharmaceutically acceptable carrier.

8. Use of a compound and a composition, characterized in that: Use of the compound of the general formula (1) according to any one of claims 1 to 6 and its optically active form, diastereomer or the composition according to claim 7 in the preparation of a drug for inhibiting CYP51.

9. The use according to claim 8, characterized in that: Use of the compound of the general formula (1) according to any one of claims 1 to 6 and its optically active form, diastereomer or the composition according to claim 7 in the preparation of antifungal drugs.

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