Ketoconazole acyltriazole analogs, methods of making and using the same

Abstract

The application belongs to the technical field of pharmaceutical chemistry, and discloses a ketoconazole acyl triazole analogue as well as a preparation method and application thereof. The structure of the ketoconazole acyl triazole analogue is shown as formula (I), and the preparation method is as follows: open reaction of ketoconazole active ester and sodium azide in N, N dimethylformamide to obtain compound 2. Compound 2 is reacted with TEMPO and chalcone derivatives in water to obtain ketoconazole acyl triazole analogue 3. The preparation method has the advantages of easy raw material and simple preparation process, and the prepared ketoconazole acyl triazole analogue has excellent antifungal activity and can be applied to the preparation of antifungal drugs.

Description

Technical Field

[0001] This invention belongs to the field of pharmaceutical chemistry technology, specifically relating to a ketoconazole acyltriazole analogue, its preparation method, and its application. Background Technology

[0002] Ketoconazole capsules are synthetic imidazole dioxane derivatives that exert their antibacterial effects by inhibiting fungal ergosterol biosynthesis and altering the composition of other lipid compounds in the cell membrane. Ketoconazole has inhibitory effects on dermatophytes such as Trichophyton, Epidermophyton, and Microsporum, as well as yeasts such as Candida.

[0003] Summary of the Invention

[0004] In view of this, the purpose of the present invention is to provide a ketoconazole acyltriazole analog, its preparation method and application, wherein the ketoconazole acyltriazole analog has excellent antifungal activity.

[0005] This invention provides a ketoconazole acyltriazole analog having the structure shown in formula (I):

[0006]

[0007] Wherein, R1 represents one of phenyl, p-chlorophenyl, and p-hydroxyphenyl, and R2 represents one of phenyl and p-fluorophenyl;

[0008] Furthermore, the ketoconazole acyltriazole derivative has a structure as shown in any one of formulas 3a-3d:

[0009]

[0010] in,

[0011] When R1 is phenyl and R2 is phenyl, the ketoconazole acyltriazole analog is a compound with the structure shown in Formula 3a.

[0012] When R1 is p-fluorophenyl and R2 is phenyl, the ketoconazole acyltriazole analog is a compound with the structure shown in Formula 3b.

[0013] When R1 is phenyl and R2 is p-chlorophenyl, this ketoconazole acyltriazole analog has the structure shown in formula 3c.

[0014] When R1 is phenyl and R2 is p-hydroxyphenyl, this ketoconazole acyltriazole analog has the structure shown in 3d.

[0015] This invention also provides a method for preparing a ketoconazole acyltriazole analog, comprising the following steps:

[0016] Compound 2 was obtained by open-air reaction of ketoconazole active ester with sodium azide in N,N-dimethylformamide. Compound 2 was then reacted with TEMPO and chalcone derivatives in water to give ketoconazole acyltriazole analog 3.

[0017] The reaction formula is as follows:

[0018]

[0019] S1. Ketoconazole active ester was dissolved in N,N-dimethylformamide, sodium azide was added, and the reaction was carried out in an open container to obtain a first reaction solution. The first reaction solution was cooled, concentrated under reduced pressure, diluted with an organic solvent, washed successively with water, washed with saturated brine, dried with MgSO4, concentrated under reduced pressure, and then column chromatography was performed to obtain a white solid compound 2, wherein the molar ratio of ketoconazole active ester to sodium azide was 1:1.2.

[0020] S2. Compound 2 was dissolved in water, and 2,2,6,6-tetramethylpiperidine oxide (TEMPO) and chalcone derivative were added sequentially. The reaction was carried out under open conditions to obtain a second reaction solution. The second reaction solution was cooled, concentrated under reduced pressure, diluted with an organic solvent, washed successively with water and saturated brine, dried over MgSO4, concentrated under reduced pressure, and then subjected to column chromatography to obtain a white solid compound 3. The molar ratio of compound 2, TEMPO, and chalcone derivative was 1:1:1.2.

[0021] The structural formula of the chalcone derivative is shown below:

[0022]

[0023] In the above formula, R1 represents one of phenyl, p-chlorophenyl, and p-hydroxyphenyl; R2 represents phenyl or p-fluorophenyl.

[0024] Furthermore, the chalcone derivative is one of chalcone, 4'-fluorochalcone, 4-hydroxychalcone, and 4-chlorochalcone.

[0025] Preferably, in step S1 of the above preparation method, the reaction temperature is 80°C and the reaction time is 12h.

[0026] Preferably, in step S2 of the above preparation method, the reaction temperature is 90°C and the reaction time is 12h.

[0027] Preferably, the organic solvent used in the above preparation method is at least one of ethyl acetate, diethyl ether, and benzene.

[0028] This invention also provides the application of the above-mentioned ketoconazole acyltriazole derivative in the preparation of antifungal drugs.

[0029] Compared with the prior art, the present invention provides a ketoconazole acyltriazole derivative containing an acyltriazole structure, which has excellent antifungal activity and can be used in the preparation of antifungal drugs. Attached Figure Description

[0030] To more clearly illustrate the technical solutions in the embodiments of the present invention or the prior art, the accompanying drawings used in the description of the embodiments or the prior art will be briefly introduced below.

[0031] Figure 1 The 1H NMR spectrum of the ketoconazole acyltriazole analog (3a) provided in Example 2 of this invention;

[0032] Figure 2 The 13C NMR spectrum of the ketoconazole acyltriazole analog (3a) provided in Example 2 of this invention;

[0033] Figure 3 This is a high-resolution mass spectrum of the ketoconazole acyltriazole analog (3a) provided in Example 2 of the present invention;

[0034] Figure 4 The 1H NMR spectrum of the ketoconazole acyltriazole analog (3b) provided in Example 3 of this invention;

[0035] Figure 5 The 13C NMR spectrum of the ketoconazole acyltriazole analog (3b) provided in Example 3 of this invention;

[0036] Figure 6 This is a high-resolution mass spectrum of the ketoconazole acyltriazole analog (3b) provided in Example 3 of the present invention;

[0037] Figure 7 The 1H NMR spectrum of the ketoconazole acyltriazole analog (3c) provided in Example 4 of this invention;

[0038] Figure 8 The 13C NMR spectrum of the ketoconazole acyltriazole analog (3c) provided in Example 4 of this invention;

[0039] Figure 9 This is a high-resolution mass spectrum of the ketoconazole acyltriazole analog (3c) provided in Example 4 of the present invention;

[0040] Figure 10 The 1H NMR spectrum of the ketoconazole acyltriazole analog (3d) provided in Example 5 of this invention;

[0041] Figure 11The 13C NMR spectrum of the ketoconazole acyltriazole analog (3d) provided in Example 5 of this invention;

[0042] Figure 12 This is a high-resolution mass spectrum of the ketoconazole acyltriazole analog (3d) provided in Example 5 of the present invention. Detailed Implementation

[0043] The technical solutions in the embodiments of the present invention will be clearly and completely described below. Obviously, the described embodiments are only some embodiments of the present invention, and not all embodiments. Based on the embodiments of the present invention, all other embodiments obtained by those skilled in the art without creative effort are within the scope of protection of the present invention.

[0044] Example 1

[0045] 200 mg (0.4 mmol) of ketoconazole active ester was dissolved in N,N-dimethylformamide (4 ml), and then 33 mg (0.48 mmol) of sodium azide was added. The mixture was reacted at 80 °C in an open container for 12 hours. The reaction solution was concentrated under reduced pressure and subjected to column chromatography (petroleum ether: ethyl acetate = 1:1) to give 130 mg of white solid, namely compound 2 (yield = 89%).

[0046] 1 H NMR(400MHz,DMSO-d6)δ7.68(d,J=2.1Hz,1H,ArH),7.50(d,J=8.5Hz,1H,ArH),7.46(s,1H,ArH),7.43(dd,J=8.5,2.1Hz,1H,ArH), 7.01(s,1H,ArH),6.84(s,1H,AH),4.55(s,2H,NCH2C),4.20(tdd,J=7.0,5.5,3.6Hz,1H),3.81(dd,J=8.5,6.8Hz,1H),3.55(dd,J=r 8.5,5.5Hz,1H),3.22(dd,J=13.0,3.6Hz,1H),3.02(dd,J=13.0,7.3Hz,1H). 13 C NMR (100MHz, DMSO) δ138.8,135.3,135.0,132.8,131.1,130.5,128.2,127.7,121.4,108.4,75.8,67.3,52.3,51.1.

[0047] Example 2

[0048] 200 mg (0.57 mmol) of compound 2 was dissolved in water, and then 97 mg (0.57 mmol) of TEMPO and 141 mg (0.69 mmol) of chalcone were added. The mixture was reacted at 90 °C for 12 hours. The reaction solution was filtered, the filtrate was concentrated under reduced pressure, and column chromatography (dichloromethane:methanol = 100:1) was performed to give 153 mg of white solid, which was compound 3a (yield 48%).

[0049] 1 H NMR (400MHz, DMSO-d6) δ8.12–8.06(m,2H,ArH),7.66(d,J=7.8Hz,2H,ArH),7.56(d,J=11.3Hz,7H,ArH),,7.40(s,2H,ArH),7.37–7.15( m,1H,ArH),6.75(s,2H,ArH),4.45–4.30(m,4H),4.21(dd,J=14.2,6.8Hz,1H),3.89(dd,J=8.6,6.1Hz,1H),3.48(dd,J=8.6,6.0Hz,1H). 13 C NMR (100MHz, DMSO) δ186.5,143.2,142.2,137.4,135.1,135.0,133.6,132.6,131.1,130.6 ,130.4,130.4,129.2,128.8,127.7,126.3,108.3,74.8,67.9,51.1,49.6..HRMS(ESI):m / z calcd for C29H23Cl2N5O3:560.1256; found:560.1250[M+H] + .

[0050] Example 3

[0051] Compound 3b of Example 2 was prepared by replacing chalcone with 4'-fluorochalcone.

[0052] 3b: Yield was 51%. 1 H NMR (400MHz, DMSO-d6) δ8.21(dd,J=8.6,5.6Hz,2H,ArH),7.69–7.50(m,6H,ArH),7.48–7.25(m,5H,ArH),6.74(d,J=16. 0Hz,2H,ArH),4.45–4.28(m,4H),4.20(dd,J=14.2,6.8Hz,1H),3.89(dd,J=8.6,6.1Hz,1H),3.48(dd,J=8.7,5.9Hz,1H). 13C NMR (100MHz, DMSO) δ186.8,159.4,142.8,142.4,138.8,137.6,135.1,135.0,133.5,132.6,132.1,131.1,13 0.5,130.4,128.8,128.2,127.7,121.2,116.1,116.0,108.2,74.8,68.0,51.1,49.5..HRMS(ESI):m / zcalcd for C29H22Cl2FN5O3:578.1162; found:578.1164[M+H] + .

[0053] Example 4

[0054] Compound 3c of Example 2 was prepared by replacing chalcone with 4-chlorochalcone.

[0055] 3c: Yield is 56%. 1 H NMR(400MHz,)δ8.27–8.21(m,2H,ArH),7.58(dd,J=7.8,3.3Hz,3H,ArH),7.50–7.43(m,4H,ArH),7.38(t,J=7.2Hz,3H,ArH),7.24(dd,J=8.4, 2.2Hz,1H,ArH),6.88(d,J=34.7Hz,2H,ArH),4.62–4.20(m,4H),4.16–4.10(m,1H),3.92(dd,J=8.7,6.2Hz,1H),3.65(dd,J=8.7,5.2Hz,1H). 13 C NMR (100MHz, CDCl3) δ186.1,143.6,141.0,136.8,136.8,136.2,133.8,133.3,132.8,131.5,131.4,130. 6,129.5,129.4,129.3,128.3,127.4,124.1,108.4,74.7,68.2,51.3,51.2,49.0..HRMS(ESI):m / zcalcd for C29H22Cl3N5O3:594.0866; found:594.0870[M+H] + .

[0056] Example 5

[0057] Compound 3d of Example was prepared by replacing chalcone with 4-hydroxychalcone according to the method of Example 2 above.

[0058] 3d: Yield was 53%. 1 H NMR(400MHz,)δ10.03(s,1H,Ar-OH),8.08–8.02(m,2H,ArH),7.68–7.62(m,2H,ArH) ,7.53(t,J=7.6Hz,2H,ArH),7.42–7.33(m,5H,ArH),6.96–6.89(m,2H,ArH),6.76(d ,J=22.5Hz,2H,ArH),4.39(dd,J=14.9,6.6Hz,3H),4.30(dd,J=14.3,4.9Hz,1H),4. 22(dd,J=14.3,6.9Hz,1H), 3.89(dd,J=8.6,6.2Hz,1H), 3.50(dd,J=8.6,6.0Hz,1H). 13 C NMR (100MHz, DMSO) δ186.8,159.4,142.8,142.4,138.8,137.6,135.1,135.0,133.5,132.6,132.1,13 1.1,130.5,130.4,128.8,128.2,127.7,121.2,116.1,108.2,74.8,68.0,51.1,49.5..HRMS(ESI):m / z calcd for C29H23Cl2N5O4:576.1205; found:576.1203[M+H] + .

[0059] The 1H NMR spectrum, 13C NMR spectrum, and high-resolution mass spectrum of ketoconazole acyltriazole analogs 3a-3d provided in Examples 2-5 are as follows: Figure 1-12 As shown.

[0060] To better understand the essence of this invention, the following pharmacological experiments demonstrating the in vitro inhibitory effects of the ketoconazole acyltriazole analogs 3a-3d provided by this invention against *Candida albicans* SC5314, *Cryptococcus neoformans*, and *Trichophyton rubrum* illustrate their novel applications in the field of antifungal drug research. The pharmacological examples provide partial activity data for representative compounds. It must be noted that the pharmacological examples of this invention are illustrative and not intended to limit the invention. Simple modifications made to this invention based on its essence are within the scope of protection claimed by this invention.

[0061] Drug Example 1: Inhibitory activity of compounds 3a-3d and ketoconazole and itraconazole against Candida albicans, Cryptococcus neoformans, and Trichophyton rubrum.

[0062] Strains and culture media: Candida albicans, Cryptococcus neoformans, and Trichophyton rubrum. Strains were stored in YPD+glycerol medium at -80°C, passaged on YPD agar plates, and activated in YPD liquid medium.

[0063] Experimental methods: The minimum inhibitory concentration (MIC) of compounds 3a-3d and the positive control drugs ketoconazole and itraconazole against the above-mentioned fungi was determined by the micro-liquid dilution method. 50 The strain was cultured on YEPD agar medium at 35°C for 24 hours, activated twice, and then the activated strain was picked with an inoculation loop, diluted with RPMI 1640 medium, and counted using a cell counting chamber. The concentration of the bacterial suspension was adjusted to the required working concentration of (2.0-2.5) × 10⁻⁶. 3 CFU / mL, freshly prepared before use. Dilute the drug stock solution to 2-fold working concentration using RPMI 1640 liquid medium. Add 100 μL of the serially diluted drug solution to columns 2-11 of a 96-well plate; column 1 is the growth control well, and column 12 is the blank control well. Add bacterial suspension to columns 1-11 to the working concentration. After incubating the plates with bacterial suspension and drug at 35°C for 48 hours, measure the OD value at 490 nm using a microplate reader. The lowest drug concentration that inhibits 50% fungal growth is taken as the final MIC. 50 The value is repeated three times.

[0064] Experimental results: The antifungal activity of the compounds in this invention was determined by the above experimental methods. The results of the in vitro antifungal activity of the compounds are shown in Table 1. Compounds 3a-3d have broad-spectrum antifungal activity.

[0065] Table 1. Antifungal activity MICs of compounds 3a-3d, ketoconazole, and itraconazole 50 (μg / mL)

[0066]

Claims

1. A ketoconazole acyltriazole analog, characterized in that, The structure of the ketoconazole acyltriazole analog is shown in the following formula: Wherein, R1 represents one of phenyl, p-chlorophenyl and p-hydroxyphenyl, and R2 represents phenyl or p-fluorophenyl.

2. The ketoconazole acyltriazole analog according to claim 1, characterized in that, The ketoconazole acyltriazole analogue has a structure as shown in any one of formulas 3a-3d: 。 3. A method for preparing the ketoconazole acyltriazole analog according to claim 1, characterized in that, The reaction formula for the preparation method is shown below: The preparation method includes the following steps: S1. Ketoconazole active ester was dissolved in N,N-dimethylformamide, sodium azide was added, and the reaction was carried out in an open container to obtain the first reaction solution. The first reaction solution was cooled, concentrated under reduced pressure, diluted with an organic solvent, washed successively with water and saturated brine, dried over MgSO4, concentrated under reduced pressure, and then subjected to column chromatography to obtain compound 2, wherein the molar ratio of ketoconazole active ester to sodium azide was 1:1.

2. S2. Compound 2 was dissolved in water, and 2,2,6,6-tetramethylpiperidine oxide and chalcone derivative were added sequentially. The reaction was carried out under open conditions to obtain a second reaction solution. The second reaction solution was cooled, concentrated under reduced pressure, diluted with an organic solvent, washed sequentially with water and saturated brine, dried over MgSO4, concentrated under reduced pressure, and then subjected to column chromatography to obtain compound 3. The molar ratio of compound 2, 2,2,6,6-tetramethylpiperidine oxide, and chalcone derivative was 1:1:1.

2. The structural formula of the chalcone derivative is shown below: ， In the above formula, R1 represents one of phenyl, p-chlorophenyl, and p-hydroxyphenyl; R2 represents phenyl or p-fluorophenyl.

4. The preparation method according to claim 3, characterized in that, In step S1, the reaction temperature is 80℃ and the reaction time is 12h.

5. The preparation method according to claim 3, characterized in that, In step S2, the reaction temperature is 90℃ and the reaction time is 12h.

6. The preparation method according to claim 3, characterized in that, The organic solvent is at least one of ethyl acetate, diethyl ether, and benzene.

7. The use of the ketoconazole acyltriazole analogue according to claim 1 in the preparation of antifungal drugs, wherein the fungus is one of Candida albicans, Cryptococcus neoformans, and Trichophyton rubrum.

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