Synthesis, Fungicidal Activity and SAR of 2-Thiazolamide/Pyrazolamide-Cyclohexylsulfonamides against Botrytis cinerea.

In order to explore more efficient sulfonamides against Botrytis cinereal, 36 novel cyclohexylsulfonamides were synthesized by N-(3-dimethylaminopropyl)-N'-ethylcarbodiimide (EDCI) and 1-hydroxybenzotriazole (HOBt) condensation reaction using chesulfamide as a lead compound, introducing thiazole and pyrazole active groups. Their structures were characterized by 1H-NMR, 13C-NMR, mass spectrum (MS), and elemental analysis. Compound III -31 was further confirmed by X-ray single crystal diffraction. The in vitro and in vivo fungicidal activities against B. cinerea were evaluated by three bioassay methods. The results of mycelial growth demonstrated that median effective concentration (EC50) values of nine compounds were close to boscalid (EC50 = 1.72 µg/mL) and procymidone (EC50 = 1.79 µg/mL) against B. cinerea (KZ-9). In the spore germination experiment, it was found that compounds III-19 and III-31 inhibited germination 93.89 and 98.00%, respectively; at 10 µg/mL, they approached boscalid (95.97%). In the tomato pot experiment, the control effects of two compounds (III-21 and III-27) were 89.80 and 87.90%, respectively, at 200 µg/mL which were significantly higher than boscalid (81.99%). The structure-activity relationship (SAR) was also discussed, which provided a valuable idea for developing new fungicides.

1. Introduction

Botrytis cinerea is one of the top 10 fungal pathogens that causes grey mould or Botrytis blight. It mainly affects vegetables and fruits, as well as a large number of flowers, shrubs, weeds and trees worldwide [1,2,3,4,5]. Many fungicides have failed to control B. cinerea because of their high risk for fungicide resistance development [6]. Therefore, it is worth developing and synthesizing new safe products that can effectively control B. cinerea.

It is a common familiarity that sulfonamide compounds have excellent biological activities not only in the field of pharmaceuticals [7,8] but also as agrochemicals [9,10,11,12], such as herbicides and fungicides. A novel sulfonamide fungicide chesulfamide [13,14] is different from other commercial fungicides; it showed excellent fungicidal activity against B. cinerea due to its unique mechanism of action, including (1) acting on mycelium cell membrane, (2) disturbing DNA, and (3) inducing disease resistance of plants [13,15]. Therefore, chesulfamide was used as a lead compound.

In the current situation, thiazolamide and pyrazolamide fungicides have become the dominant fungicide varieties, such as Ethaboxam, Thfluzamide [16], Bixafen [17], Sedaxane [18], Fluxapyroxad [19], and Isopyrazam [20] (Figure 1). Among them, the alkyl-thiazole/pyrazole group was selected. In the previous study, our laboratory placed emphasis on the study of chesulfamide and synthesized a series of 2-pyridyl, 2-phenyl 2-isothiazolyl, 2-thiazolyl and 2-pyrazolyl cyclohexylsulfonamides against B. cinerea [21,22,23,24,25] (Scheme 1a). Phenylthiazole and phenylpyrazole active substructures were screened out. Hence, the fungicidal activities of two substituted thiazoles/pyrazoles compounds were contrasted.

Figure 1

Chemical structure containing thiazole and pyrazole fungicides.

Scheme 1

Design ideas for the target compounds.

In this study, we used chesulfamide as a lead compound, reducing the carbonyl group to an amino group [26,27], and the EDCI/HOBt condensation method [24,25], interfacing with the pyrazole carboxylic acid and thiazole carboxylic acid. In order to improve the fungicidal activities against B. cinerea, the benzene ring (linked to NH) was optimized, which was different from the structure of 2-trifluoromethyl-4-chlorobenzene before. Moreover, 18 novel 2-thiazolamide cyclohexylsulfonamides and 18 novel 2-pyrazolamide cyclohexylsulfonamides were designed and synthesized (Scheme 1b). The in vivo and in vitro fungicidal activities against B. cinerea were evaluated and the structure–activity relationship was analyzed. The synthetic route of the target compounds was shown in Scheme 2.

Scheme 2

Synthetic route of the target compounds III.

2. Results

2.1. Chemistry

The corresponding aminocyclohexylsulfonamide (I), HOBt and EDCI in the presence of triethylamine (Et3N) in dichloromethane (CH2Cl2) as solvent were allowed to react with different carboxylic acids (II) at 0 °C. Finally, 36 novel cyclohexylsulfonamides compounds III were synthesized.

The structure of compound III-31 was analyzed by X-ray single crystal diffraction. The structure (CCDC: 1917351) was shown in Figure 2a and available in Supplementary Data. X-ray diffraction provided a relative configuration of chiral centers, hence it proved the cis-configuration at the cyclohexane ring. Probably, the compound III-31 was racemates, which mixed both enantiomers (“C7”R, “C12”S and “C7”S, “C12”R). A compound reported (Figure 2b) [26] with a similar structure showed that there were two intramolecular hydrogen bonds in the structure, N1-H…O3 and N2-H…O2. On the contrary, in our study, compound III-31 had no intramolecular hydrogen bonds, but had intermolecular hydrogen bonds, with N1 in one molecule being linked to O3 in another molecule, eventually forming the chain structure (Figure 2c).

Figure 2

X-ray crystal structures.

Additionally, in this work, the chemical structures of all the synthesized compounds were characterized by nuclear magnetic resonance (NMR), mass spectrum (MS) and elemental analysis. The 1H NMR and 13C NMR spectra are available in Supplementary Data. In the 1H NMR spectra (600 MHz, DMSO-d6) of compound III-28 (Figure 3a), the protons on the benzene ring were revealed in a low field in the range of δH 7.4 to 7.6 ppm, while cyclohexyl and methyl group gave signals in the range of δH 1.0 to 5.0 ppm. However, pyrazole-H and SO2NH appeared at δH 8.36 ppm and 10.13 ppm, respectively. The carboxamide proton of NH–C=O showed doublet (d) at δH 7.67 ppm, J = 8.4 Hz. The signal of proton of CH-SO2 at δH 4.64 ppm revealed only coupling constants less than 9 Hz, indicating the equatorial position of this proton. Analogously, the axial orientation of the proton on CH-N was revealed by its splitting to doublet of triplets (dt) at δH 3.52 ppm with one diaxial coupling of J = 11.4 Hz and two axial equatorial couplings of J = 3.6 Hz. The splitting pattern of protons on CHF2 is very characteristic. Generally, the proton of CHF2 showed triplet (t) at δH 7.26ppm, J = 54 Hz.

Figure 3

Structure elucidation of compounds III.

In the 13C-NMR spectra (151 MHz, DMSO-d6) of compound III-28 (Figure 3b), the signals in the range of δC 0 to 70 ppm were assigned to methyl and cyclohexyl groups. Benzene ring carbons and most of the pyrazole carbons resonated in the range of δC 112 to 140 ppm and the signal at δC 161 ppm belonged to C=O. The difluoromethyl group caused the triplet at δC 110.16 ppm with 1JFC = 234.05 Hz, while the adjacent carbon signal at δC 144.78 ppm is split into a triplet due to 2JFC = 22.80 Hz.

In the 13C-NMR spectra (151 MHz, DMSO-d6) of compound III-25 (Figure 3c), there were very interesting phenomena due to seven fluorine atoms. Carbon signals of C-F in the F-Ph showed a doublet at δC 162.10 ppm, 1JFC = 246.43 Hz, while carbon signals of C-F in 2,4,5-trifluorobenzene, which appeared in the range of δC 145 to 151 ppm, showed 1JFC couplings of 245.38, 244.77, 239.03 Hz, respectively, and further long-range C-F coupling. The carbon signals of CF3 appeared as a quartet at δC 119.61 ppm with 1JFC = 270.74 Hz, while the quartet of the adjacent carbon at δC 129.96 ppm showed the 2JFC coupling of 39.26 Hz.

2.2. Biological Assay and Structure—Activity Relationship Study

In order to screen out active compounds quickly and correctly, the target compounds were tested by an in vitro mycelium growth inhibition assay and the spore germination method. Then, active compounds were tested in tomato pot experiments.

2.2.1. In Vitro Fungicidal Activity against B. cinerea

The germicidal virulence test of 36 target compounds was tested against the B. cinerea strain (KZ-9), with boscalid and procymidone as standard fungicides. The EC50 values of 17 compounds were lower than 10 μg/mL. Compounds III-19 and III-27 showed better in vitro fungicidal activity against KZ-9, with the EC50 values of 1.99 and 2.04 μg/mL (Table 1). Some compounds showed excellent fungicidal activity against KZ-9 and were rescreened out by another B. cinerea strain (CY-09). The EC50 value of compound III-21 against CY-09 was 2.33 μg/mL, which showed its excellent fungicidal activity (Table 2).

Table 1

The EC50 values of target compounds III against B. cinerea (KZ-9) in vitro.

Compound	R1	R2		EC50 (μg/mL)
III-1	2-F			4.96
III-2	3-F			2.32
III-3	2-Cl			2.99
III-4	3-Cl			2.33
III-5	4-Cl			21.12
III-6	2-Br			18.70
III-7	3-Br			89.45
III-8	4-Br			18.79
III-9	2,4,5-F			2.10
III-10	2-F			37.00
III-11	3-F			3.65
III-12	2-Cl			44.3
III-13	3-Cl			>100
III-14	4-Cl			17.97
III-15	2-Br			>100
III-16	3-Br			25.78
III-17	4-Br			10.01
III-18	3-CN			44.72
III-19	3-F			1.99
III-20	2-Cl			5.94
III-21	4-Cl			2.11
III-22	2-Br			7.01
III-23	3-Br			3.27
III-24	4-Br			2.56
III-25	2,4,5-F			5.76
III-26	3-Cl			14.75
III-27	2-F			2.04
III-28	3-CN			>100
III-29	2-F			31.47
III-30	2-Br			17.53
III-31	2-Cl			9.93
III-32	3-F			44.91
III-33	4-Br			21.16
III-34	3-Br			18.63
III-35	2,4,5-F			6.47
III-36	4-Cl			3.37
boscalid	/	/		1.72
procymidone	/	/		1.79

Table 2

The EC50 values of selected target compounds III against B. cinerea (CY-09) in vitro.

Compound	R1	R2	EC50 (μg/mL)
III-2	3-F		>100
III-3	2-Cl		>100
III-4	3-Cl		8.01
III-9	2,4,5-F		>100
III-19	3-F		12.64
III-21	4-Cl		2.33
III-23	3-Br		7.66
III-27	2-F		5.70
boscalid	/	/	2.25
procymidone	/	/	5.19

The results represented in Table 3 indicated that several compounds can inhibit the spore germination of B. cinerea at the concentration of 10 μg/mL. Among these, compounds III-19 and III-31 showed superb activity on spore germination, which had higher inhibition rates (93.89 and 98.0%) were comparable to boscalid (95.97%).

Table 3

Inhibition rate of target compounds III for spore germination at 10 μg/mL.

Compound	Inhibition Rate%	Compound	Inhibition Rate%
III-1	24.23	III-19	93.89
III-2	72.34	III-20	36.80
III-3	75.98	III-21	75.00
III-4	21.01	III-22	67.79
III-5	41.00	III-23	42.00
III-6	61.78	III-24	67.86
III-7	49.33	III-25	70.94
III-8	17.69	III-26	30.00
III-9	26.44	III-27	80.82
III-10	24.37	III-28	60.00
III-11	34.89	III-29	39.57
III-12	36.48	III-30	42.58
III-13	50.77	III-31	98.00
III-14	61.34	III-32	77.66
III-15	10.10	III-33	41.70
III-16	14.36	III-34	42.57
III-17	73.77	III-35	37.87
III-18	72.09	III-36	35.82
boscalid	95.97	/	/

2.2.2. In Vivo Fungicidal Activity against B. cinerea

After that, the in vivo control effect of active compounds against B. cinerea on tomato leaves were tested at 200 μg/mL. The results showed that the control effects of compounds III-19, III-21 and III-27 were 78.00, 89.80 and 87.90%, respectively, which were similar to boscalid (81.99%). The three compounds showed excellent activities in in vivo tests (Table 4).

Table 4

Control efficiency of specific target compounds against B. cinerea in tomato pot experiments.

Compd.	Control Effect%
III-2	14.15
III-3	52.33
III-19	78.00
III-21	89.80
III-27	87.90
boscalid	81.99

Accordingly, by analyzing the experimental results, the structure–activity relationship (SAR) was summarized as follows: (1) Compounds III-10–III-18 had the same alkyl-thiazole structure as that of Thifluzamide while compounds III-1–III-9 had phenylthiazole structure. It was noted that compounds III-1–III-9 showed significantly improved fungicidal activities. In addition, compounds containing pyrazoles revealed the same results; it showed that fungicidal activities of compounds III-19–III-27 were better than compounds III-28–III-36. In total, compounds with benzene ring-substituted thiazole or pyrazole active groups showed more highly fungicidal activities. (2) For 2-thiazolamide-cyclohexylsulfonamides (III-1–III-18), the activity was significantly increased when the benzene ring (linked to NH) contained fluorine-containing groups (2-F, 3-F and 2,4,5-F), such as compounds III-1, III-2, III-9 and III-11. When the benzene ring (linked to NH) contained chlorine, the phenylthiazole compounds III-3 and III-4 showed excellent fungicidal activity. However, fungicidal activity was mediocre on the benzene ring containing bromine. (3) For 2-pyrazolamide-cyclohexylsulfonamides (III-19–III-36), the activity was higher when the benzene ring (linked to NH) contained fluorine-containing groups (2-F, 3-F), such as phenylpyrazole compounds III-19 and III-27, but, if the substituent was a 2,4,5-fluorine, the fungicidal activity slightly decreased. Additionally, when the benzene ring (linked to NH) contained 4-chlorine, the compounds III-21 and III-36 showed excellent activity. When the benzene ring (linked to NH) contained bromine, the phenylpyrazole compounds III-23 and III-24 showed excellent activity. 4) It could be seen that the compounds III containing pyrazole substituent showed higher fungicidal activity than thiazole substituent.

3. Materials and Methods

3.1. Materials and Instrumentation

The reagents and solvents were commercially available for analytical reagent (AR) grade and were used as received or were dried prior to use, as needed. The melting point was measured by X-5 binocular microscope melting point apparatus (Beijing Tech Instruments Co. Ltd., Beijing, China). Using dimethyl sulfoxide (DMSO-d6) as solvent and tetramethylsilane (TMS) as the internal standard, 1H-NMR spectra were recorded on 300 MHz and 600 MHz spectrometers and 13C-NMR spectra were recorded on 75 MHz and 151 MHz spectrometers (Bruker, Karlsruhe, Germany). MS data were obtained on the 7000C Triple Quad GC/MS and 6460 Triple Quad LC/MS Mass Spectrometers (Agilent Technologies, Santa Clara, CA, USA). Elemental analyses were determined on a Vario EL III elemental analyser (Elementar Analysensysteme GmbH, Frankfurt, Germany).

3.2. Synthesis

3.2.1. Synthesis of N-Substituted Phenyl-2-Aminocyclohexylsulfonamides I

The synthesis of N-substituted phenyl-2-aminocyclohexylsulfonamide I was done according to the reported method [10,21,27]. In this paper, 2-trifluoromethyl-4-chloroaniline in the reference was replaced by different substituted aniline, but the experimental methods were consistent. The yield of the compounds I was 40–75%.

3.2.2. Synthesis of Target Compounds III

Under nitrogen atmosphere, carboxylic acid II (3mmol), EDCI (3.3 mmol), HOBT (3.3 mmol) and Et3N (1.8 mmol) were placed in a three-necked flask with 40 mL CH2Cl2, and stirred for 2 h at 0 °C; then, compound I (2.4 mmol) was added to the flask and allowed to react for 3 h at 0 °C. The reaction was monitored by thin-layer chromatography (TLC) (all reactions could be completed in 3 h) and, on completion of the reaction, the mixture was washed with saturated NaHCO3 solution and water, respectively. Then, it was dried over anhydrous Na2SO4, filtered and evaporated on rotavapor in vacuum. Subsequently, crude products III-1–III-18 were purified by silica gel column chromatography [V (CH2Cl2): V (EA) = 3:1] and crude products III-19–III-36 were purified by silica gel column chromatography [V (PE): V (EA) = 3:1]. Finally, products were recrystallized with the dichloromethane/petroleum ether to obtain pure target compounds.

N-(2-fluorophenyl)-2-[4-methyl-2-(4-trifluoromethylphenyl)-5-thiazolecarboxylamino]-cyclohexylsulfonamide III-1. White solid, yield: 85%. mp 156.8~157.7 °C. 1H-NMR (300 MHz, DMSO-d): δ (ppm) 9.63 (s, 1H, NH-SO2), 8.22–7.17 (m, 9H, NH–C=O + C6H4 + C6H4), 4.68 (dd, J = 7.5, 3.3 Hz, 1H, CH–SO2), 3.33–3.30 (m, 1H, CH–N) (overlap with water peak), 2.63 (s, 3H, CH3), 2.16–1.27 (m, 8H, 4CH2). 13C-NMR (75 MHz, DMSO-d6): δ (ppm) 162.86 (d, 1JFC = 237.0 Hz, F-Ph), 157.33, 154.92, 154.08, 141.65, 136.46, 130.83 (d, 2JFC = 32.25 Hz), 128.81, 127.43, 127.37, 126.86, 126.81, 126.77, 125.28, 125.17, 116.63, 116.37, 60.21, 45.96, 30.41, 24.17, 22.07, 19.89, 17.31. Electron impact mass spectrometry (EIMS), m/z 541.58 (M). Elemental analysis for C24H23F4N3O3S2: found C 53.23, H 4.28, N 7.76; calcd C 53.02, H 4.12, N 7.98.

N-(3-fluorophenyl)-2-[4-methyl-2-(4-trifluoromethylphenyl)-5-thiazolecarboxylamino]-cyclohexylsulfonamide III-2. White solid, yield: 82%. mp 160.6~161.7 °C. 1H-NMR (300 MHz, DMSO-d): δ (ppm) 10.06 (s, 1H, NH-SO2), 8.21-6.87 (m, 9H, NH–C=O + C6H4 + C6H4), 4.65 (dd, J = 6.9, 3.6 Hz, 1H, CH–SO2), 3.46 (dt, J = 11.1, 3.3 Hz, 1H, CH–N), 2.64 (s, 3H, CH3), 2.15–1.23 (m, 8H, 4CH2). 13C-NMR (151 MHz, DMSO-d6): δ (ppm) 164.41, 162.91 (d, 1JFC = 243.11 Hz, F-Ph), 161.11, 155.04, 140.56 (d, 3JFC = 10.57 Hz), 136.48, 131.51 (d, 3JFC = 10.57 Hz), 130.84 (q, 2JFC = 31.71 Hz), 128.71, 127.37, 126.76, 124.40 (q, 1JFC = 273.31 Hz, CF3), 115.19, 110.56, 110.42, 106.26, 106.10, 60.39, 46.14, 30.26, 23.86, 22.26, 20.04, 17.33. EIMS, m/z 541.58 (M). Elemental analysis for C24H23F4N3O3S2: found C 53.23, H 4.28, N 7.76; calcd C 53.51, H 4.07, N 7.88.

N-(2-chlorophenyl)-2-[4-methyl-2-(4-trifluoromethylphenyl)-5-thiazolecarboxylamino]-cyclohexylsulfonamide III-3. White solid, yield: 84%. mp 117.0~118.4 °C. 1H-NMR (300 MHz, DMSO-d): δ (ppm) 9.46 (s, 1H, NH-SO2), 8.19–7.21 (m, 9H, NH–C=O + C6H4 + C6H4), 4.71 (dd, J = 7.2, 3.0 Hz, 1H, CH–SO2), 3.41–3.37 (m, 1H, CH–N) (overlap with water peak), 2.63 (s, 3H, CH3), 2.09–1.23 (m, 8H, 4CH2). 13C-NMR (75 MHz, DMSO-d6): δ (ppm) 164.44, 161.36, 154.90, 136.46, 134.56, 131.03, 130.61, 130.32, 128.81, 128.70, 128.33, 128.24, 127.53, 127.38, 126.82, 126.76, 126.70, 62.37, 45.93, 30.52, 24.30, 22.16, 19.84, 17.31. EIMS, m/z 558.03 (M). Elemental analysis for C24H23ClF3N3O3S2: found C 51.66, H 4.15, N 7.53; calcd C 51.78, H 4.02, N 7.66.

N-(3-chlorophenyl)-2-[4-methyl-2-(4-trifluoromethylphenyl)-5-thiazolecarboxylamino]-cyclohexylsulfonamide III-4. White solid, yield: 85%. mp 116.5~117.8 °C. 1H-NMR (300 MHz, DMSO-d): δ (ppm) 10.12 (s, 1H, NH-SO2), 8.31–7.36 (m, 9H, NH–C=O + C6H4 + C6H4), 4.71 (dd, J = 7.2, 3.0 Hz, 1H, CH–SO2), 3.54–3.45 (m, 1H, CH–N) (overlap with water peak), 2.61 (d, J = 9.0 Hz, 3H, CH3), 2.13–1.31 (m, 8H, 4CH2). 13C-NMR (75 MHz, DMSO-d6): δ(ppm) 164.40, 161.15, 154.96, 136.45, 134.65, 134.30, 129.75, 128.72, 127.37, 127.12, 126.93, 126.76, 122.59, 117.29, 109.99, 109.34, 109.14, 62.21, 45.93, 30.50, 24.35, 22.18, 19.83, 17.31. EIMS, m/z 558.03 (M). Elemental analysis for C24H23ClF3N3O3S2: found C 51.66, H 4.15, N 7.53; calcd C 51.79, H 4.01, N 7.62.

N-(4-chlorophenyl)-2-[4-methyl-2-(4-trifluoromethylphenyl)-5-thiazolecarboxylamino]-cyclohexylsulfonamide III-5. White solid, yield: 86%. mp 198.0~198.9 °C. 1H-NMR (300 MHz, DMSO-d): δ (ppm) 9.93 (s, 1H, NH-SO2), 8.20–7.21 (m, 9H, NH–C=O + C6H4 + C6H4), 4.63 (dd, 1H, J = 5.4, 2.1 Hz, 1H, CH–SO2), 3.37 (dt, J = 11.4, 3.3 Hz, 1H, CH–N), 2.63 (s, 3H, CH3), 2.08–1.24 (m, 8H, 4CH2). 13C-NMR (151 MHz, DMSO-d6): δ (ppm) 164.52, 161.16, 155.05, 137.57, 136.43, 130.86 (q, 2JFC = 33.22 Hz), 129.70 (two carbon atoms), 128.67, 128.23, 127.68, 127.38, 126.79 (d, 3JFC = 4.53 Hz), 124.38 (q, 1JFC = 273.31 Hz, CF3) 121.48 (two carbon atoms), 120.79, 60.30, 46.10, 30.27, 23.88, 22.22, 19.97, 17.31. EIMS, m/z 558.03 (M). Elemental analysis for C24H23ClF3N3O3S2: found C 51.66, H 4.15, N 7.53; calcd C 51.52, H 3.99, N 7.81.

N-(2-bromophenyl)-2-[4-methyl-2-(4-trifluoromethylphenyl)-5-thiazolecarboxylamino]-cyclohexylsulfonamide III-6. White solid, yield: 83%. mp 112.3~113.3 °C. 1H-NMR (300 MHz, DMSO-d): δ (ppm) 9.35 (s, 1H, NH-SO2), 8.19–7.14 (m, 9H, NH–C=O + C6H4 + C6H4), 4.72 (dd, J = 7.2, 2.7 Hz, 1H, CH–SO2), 3.44–3.39 (m, 1H, CH–N) (overlap with water peak), 2.63 (s, 3H, CH3), 2.10–1.39 (m, 8H, 4CH2). 13C-NMR (151 MHz, DMSO-d6): δ (ppm) 164.44, 161.34, 154.91, 136.46, 135.94, 133.56, 130.84 (q, 2JFC = 31.71 Hz), 130.10, 128.90, 128.82, 128.02, 127.95, 127.38, 126.78, 124.40 (q, 1JFC = 273.31 Hz, CF3), 119.41, 99.99, 62.66, 45.95, 30.56, 24.32, 22.19, 19.82, 17.32. EIMS, m/z 602.49 (M). Elemental analysis for C24H23BrF3N3O3S2: found C 47.85, H 3.85, N 6.97; calcd C 47.62, H 4.01, N 7.11.

N-(3-bromophenyl)-2-[4-methyl-2-(4-trifluoromethylphenyl)-5-thiazolecarboxylamino]-cyclohexylsulfonamide III-7. White solid, yield: 84%. mp 187.2~188.3 °C. 1H-NMR (300 MHz, DMSO-d): δ (ppm) 10.02 (s, 1H, NH-SO2), 8.19–7.22 (m, 9H, NH–C=O + C6H4 + C6H4), 4.65 (dd, J = 5.7, 2.4 Hz, 1H, CH–SO2), 3.45 (dt, J = 11.1, 3.6 Hz, 1H, CH–N), 2.64 (s, 3H, CH3), 2.14–1.28 (m, 8H, 4CH2). 13C-NMR (151 MHz, DMSO-d): δ (ppm) 164.41, 161.10, 155.08, 140.40, 139.85, 136.49, 134.55, 131.75, 130.84 (q, 2JFC = 31.71 Hz), 128.67, 127.37, 126.81, 126.68, 124.40 (q, 1JFC = 271.8 Hz, CF3), 122.50, 121.77, 118.24, 60.59, 46.16, 30.25, 23.84, 22.30, 20.06, 17.36. EIMS, m/z 602.49 (M). Elemental analysis for C24H23BrF3N3O3S2: found C 47.85, H 3.85, N 6.97; calcd C 48.02, H 3.66, N 7.12.

N-(4-bromophenyl)-2-[4-methyl-2-(4-trifluoromethylphenyl)-5-thiazolecarboxylamino]-cyclohexylsulfonamide III-8. White solid, yield: 88%. mp 209.5~210.6 °C. 1H-NMR (300 MHz, DMSO-d): δ (ppm) 9.95 (s, 1H, NH-SO2), 8.19–7.16 (m, 9H, NH–C=O + C6H4 + C6H4), 4.64 (dd, J = 6.9, 3.0 Hz, 1H, CH–SO2), 3.41–3.39 (m, 1H, CH–N) (overlap with water peak), 2.64 (s, 3H, CH3), 2.09–1.28 (m, 8H, 4CH2). 13C-NMR (151 MHz, DMSO-d): δ (ppm) 164.43, 161.11, 155.01, 138.09, 136.48, 132.61 (two carbon atoms), 130.84 (q, 2JFC = 31.71 Hz), 128.75, 127.38, 126.81, 126.78, 124.40 (q, 1JFC = 271.8 Hz, CF3), 121.95, 121.69 (two carbon atoms), 116.16, 60.27, 46.08, 30.29, 23.92, 22.22, 19.98, 17.35. EIMS, m/z 602.49 (M). Elemental analysis for C24H23BrF3N3O3S2: found C 47.85, H 3.85, N 6.97; calcd C 47.69, H 4.00, N 7.12.

N-(2,4,5-trifluorophenyl)-2-[4-methyl-2-(4-trifluoromethylphenyl)-5-thiazolecarboxylamino]-cyclohexylsulfonamide III-9. White solid, yield: 86%. mp 167.0~168.0 °C. 1H-NMR (300 MHz, DMSO-d): δ (ppm) 9.86 (s, 1H, NH-SO2), 8.19–7.47 (m, 7H, NH–C=O + C6H2 + C6H4), 4.66 (dd, J = 6.9, 2.1 Hz, 1H, CH–SO2), 3.48-3.42 (m, 1H, CH–N) (overlap with water peak), 2.62(s, 3H, CH3), 2.12-1.44 (m, 8H, 4CH2). 13C-NMR (151 MHz, DMSO-d): δ (ppm) 170.78, 164.45, 161.23, 155.00, 151.23 (dd, 1JFC = 243.11, 9.82 Hz, F-Ph), 147.03 (dt, JFC = 248.24, 11.33 Hz, F-Ph), 146.07 (dd, 1JFC = 245.68, 13.44 Hz, F-Ph) 136.46, 130.85 (q, 2JFC = 30.2 Hz), 128.66, 127.37, 126.76, 124.39 (q, 1JFC = 271.8 Hz, CF3),122.10, 114.87,114.83, 106.77 (dd, 2JFC = 26.73, 22.20 Hz), 60.21, 45.85, 30.40, 24.15, 21.21, 17.28, 14.53. EIMS, m/z 577.56 (M). Elemental analysis for C24H21F6N3O3S2: found C 49.91, H 3.67, N 7.28; calcd C 50.11, H 3.82, N 7.10.

N-(2-fluorophenyl)-2-(2-methyl-4-trifluoromethyl-5-thiazolecarboxylamino]-cyclohexylsulfonamide III-10. White solid, yield: 88%. mp 175.6~176.8 °C. 1H-NMR (300 MHz, DMSO-d): 9.52 (s, 1H, NH–SO2), 8.91 (d, J = 8.7 Hz, 1H, NH–C=O), 7.46–7.18 (m, 4H, C6H4), 4.68 (dd, J = 8.4, 2.7 Hz, 1H, CH–SO2), 3.25 (dt, J = 12.3, 3.3 Hz, 1H, CH–N), 2.73 (s, 3H, CH3), 2.00–1.24 (m, 8H, 4CH2). 13C-NMR (75 MHz, DMSO-d): δ (ppm) 167.52, 158.95, 155.83 (d, 1JFC = 243.83 Hz, F-Ph), 138.64 (d, 2JFC = 35.78 Hz), 135.09, 127.43 (d, 3JFC = 7.43 Hz), 127.09, 125.21, 120.79 (q, 1JFC = 269.63 Hz, CF3), 116.62, 116.36, 61.37, 45.37, 30.61, 24.29, 21.59, 19.48, 18.97. EIMS, m/z 465.48 (M). Elemental analysis for C18H19F4N3O3S2: found C 46.45, H 4.11, N 9.03; calcd C 46.29, H 4.32, N 9.21.

N-(3-fluorophenyl)-2-(2-methyl-4-trifluoromethyl-5-thiazolecarboxylamino]-cyclohexylsulfonamide III-11. White solid, yield: 81%. mp 159.0~160.1 °C. 1H-NMR (300 MHz, DMSO-d): 9.99 (s, 1H, NH–SO2), 8.95 (d, J = 9.0 Hz, 1H, NH–C=O), 7.39–6.86 (m, 4H, C6H4), 4.69 (dd, J = 7.8, 2.4 Hz, 1H, CH–SO2), 3.38–3.35 (m, 1H, CH–N) (overlap with water peak), 2.74 (s, 3H, CH3), 2.05–1.20 (m, 8H, 4CH2). 13C-NMR (75 MHz, DMSO-d6): δ (ppm) 167.49, 162.89 (d, 1JFC = 241.20 Hz, F-Ph), 158.84, 140.64 (d, 3JFC = 10.65 Hz), 138.71 (d, 2JFC = 35.63 Hz), 135.11, 131.49 (d, 3JFC = 9.45 Hz), 115.17, 110.42 (d, 2JFC = 20.70 Hz), 106.30, 105.96, 60.45, 45.46, 30.49, 24.04, 21.75, 19.51, 18.97. EIMS, m/z 465.48 (M). Elemental analysis for C18H19F4N3O3S2: found C 46.45, H 4.11, N 9.03; calcd C 46.57, H 3.96, N 8.91.

N-(2-chlorophenyl)-2-(2-methyl-4-trifluoromethyl-5-thiazolecarboxylamino]-cyclohexylsulfonamide III-12. White solid, yield: 85%. mp 188.8~189.8 °C. 1H-NMR (300 MHz, DMSO-d6): δ (ppm) 9.34 (s, 1H, NH–SO2), 8.92 (d, J = 9.0 Hz, 1H, NH–C=O), 7.53–7.21 (m, 4H, C6H4), 4.70 (dd, J = 8.7, 2.1 Hz, 1H, CH–SO2), 3.29 (dt, J = 11.1, 3.9 Hz, 1H, CH–N), 2.73 (s, 3H, CH3), 2.04–1.24 (m, 8H, 4CH2). 13C-NMR (75 MHz, DMSO-d6): δ (ppm) 167.53, 159.02, 135.13, 134.57, 130.30, 128.50, 128.30, 127.75, 127.62, 122.58, 118.99, 62.38, 45.38, 30.71, 24.41, 21.66, 19.42, 18.97. EIMS, m/z 481.93 (M). Elemental analysis for C18H19ClF3N3O3S2: found C 44.86, H 3.97, N 8.72; calcd C 44.69, H 3.75, N 8.94.

N-(3-chlorophenyl)-2-(2-methyl-4-trifluoromethyl-5-thiazolecarboxylamino]-cyclohexylsulfonamide III-13. White solid, yield: 82%. mp 160.5~161.8 °C. 1H-NMR (300 MHz, DMSO-d6): δ (ppm) 10.10 (d, J = 31.8 Hz, 1H, NH–SO2), 8.89 (dd, J = 14.7, 8.4 Hz, 1H, NH–C=O), 7.86–7.37 (m, 4H, C6H4), 4.72-4.10 (m, 1H, CH–SO2), 3.36–3.28 (m, 1H, CH–N) (overlap with water peak), 2.71 (d, J = 10.2 Hz, 3H, CH3), 2.08-1.23 (m, 8H, 4CH2). 13C-NMR (75 MHz, DMSO-d6): δ (ppm) 167.82, 158.86, 157.93, 139.93, 134.58, 134.27, 127.23, 127.01, 117.47, 117.33, 109.52, 63.96, 48.44, 32.34, 26.84, 24.06, 21.70, 19.01. EIMS, m/z 481.93 (M). Elemental analysis for C18H19ClF3N3O3S2: found C 44.86, H 3.97, N 8.72; calcd C 45.01, H 4.20, N 8.59.

N-(4-chlorophenyl)-2-(2-methyl-4-trifluoromethyl-5-thiazolecarboxylamino]-cyclohexylsulfonamide III-14. White solid, yield: 84%. mp 184.0~185.1 °C. 1H-NMR (300 MHz, DMSO-d6): δ (ppm) 9.87 (s, 1H, NH–SO2), 8.93 (d, J = 9.0 Hz, 1H, NH–C=O), 7.41–7.21 (m, 4H, C6H4), 4.67 (dd, J = 8.7, 2.7 Hz, 1H, CH–SO2), 3.27 (dt, J = 12.0, 3.9 Hz, 1H, CH–N), 2.74 (s, 3H, CH3), 2.09-1.18 (m, 8H, 4CH2). 13C-NMR (151 MHz, DMSO-d6): δ (ppm) 167.50, 158.84, 138.71 (q, 2JFC = 35.79 Hz), 137.73, 135.10, 129.70 (two carbon atoms), 128.08, 121.37 (two carbon atoms), 120.80 (q, 1JFC = 271.50 Hz, CF3), 60.28, 45.45, 30.53, 24.08, 21.71, 19.48, 18.99. EIMS, m/z 481.93 (M). Elemental analysis for C18H19ClF3N3O3S2: found C 44.86, H 3.97, N 8.72; calcd C 44.98, H 3.77, N 8.50.

N-(2-bromophenyl)-2-(2-methyl-4-trifluoromethyl-5-thiazolecarboxylamino]-cyclohexylsulfonamideIII-15. White solid, yield: 85%. mp 173.9~174.0 °C. 1H-NMR (300 MHz, DMSO-d6): δ (ppm) 9.24 (s, 1H, NH–SO2), 8.92 (d, J = 9.3 Hz, 1H, NH–C=O), 7.69–7.15 (m, 4H, C6H4), 4.71 (dd, J = 8.7, 2.1 Hz, 1H, CH–SO2), 3.32–3.28 (m, 1H, CH–N) (overlap with water peak), 2.73 (s, 3H, CH3), 2.10–1.29 (m, 8H, 4CH2). 13C-NMR (75 MHz, DMSO-d6): δ (ppm) 167.51, 159.00, 138.59 (q, 2JFC = 35.78 Hz), 135.95, 135.12, 133.53, 128.86, 128.19 (d, 3JFC = 13.28 Hz), 120.79 (q, 1JFC = 269.70 Hz, CF3), 119.65, 109.99, 62.65, 45.39, 30.75, 24.45, 21.71, 19.40, 18.97. EIMS, m/z 526.39 (M). Elemental analysis for C18H19BrF3N3O3S2: found C 41.07, H 3.64, N 7.98; calcd C 40.95, H 3.51, N 8.14.

N-(3-bromophenyl)-2-(2-methyl-4-trifluoromethyl-5-thiazolecarboxylamino]-cyclohexylsulfonamideIII-16. White solid, yield: 86%. mp 196.0~196.9 °C. 1H-NMR (300 MHz, DMSO-d6): δ (ppm) 9.96 (s, 1H, NH–SO2), 8.94 (d, J = 9.0 Hz, 1H, NH–C=O), 7.40–7.21 (m, 4H, C6H4), 4.68 (dd, J = 8.1, 2.1 Hz, 1H, CH–SO2), 3.32–3.31 (m, 1H, CH–N) (overlap with water peak), 2.74 (s, 3H, CH3), 2.08–1.24 (m, 8H, 4CH2). 13C-NMR (151 MHz, DMSO-d6): δ (ppm) 167.49, 158.84, 140.50, 138.71 (q, 2JFC = 35.79 Hz), 135.10, 131.75, 126.62, 122.48, 121.76, 120.80 (q, 1JFC = 271.50 Hz, CF3), 118.20, 60.68, 45.48, 30.51, 24.03, 21.77, 19.50, 18.98. EIMS, m/z 526.39 (M). Elemental analysis for C18H19BrF3N3O3S2: found C 41.07, H 3.64, N 7.98; calcd C 41.26, H 3.81, N 8.06.

N-(4-bromophenyl)-2-(2-methyl-4-trifluoromethyl-5-thiazolecarboxylamino]-cyclohexylsulfonamide III-17. White solid, yield: 83%. mp 198.1~199.1 °C. 1H-NMR (300 MHz, DMSO-d6): δ (ppm) 9.88 (s, 1H, NH–SO2), 8.94 (d, J = 9.0 Hz, 1H, NH–C=O), 7.53–7.15 (m, 4H, C6H4), 4.67 (dd, J = 8.1, 2.4 Hz, 1H, CH–SO2), 3.28 (dt, J = 12.0, 3.6 Hz, 1H, CH–N), 2.74 (s, 3H, CH3), 2.07-1.15 (m, 8H, 4CH2). 13C-NMR (151 MHz, DMSO-d6): δ (ppm) 167.50, 158.85, 138.72 (q, 2JFC = 35.79 Hz), 138.19, 135.10, 132.59 (two carbon atoms), 121.66 (two carbon atoms), 120.80 (q, 1JFC = 271.20 Hz, CF3), 116.07, 60.29, 45.44, 30.53, 24.08, 21.71, 19.48, 18.98. EIMS, m/z 526.39 (M). Elemental analysis for C18H19BrF3N3O3S2: found C 41.07, H 3.64, N 7.98; calcd C 40.89, H 3.47, N 7.69.

N-(3-cyanophenyl)-2-(2-methyl-4-trifluoromethyl-5-thiazolecarboxylamino]-cyclohexylsulfonamide III-18. White solid, yield: 88%. mp 196.0~197.0 °C. 1H-NMR (300 MHz, DMSO-d6): δ (ppm) 10.17 (s, 1H, NH–SO2), 8.95 (d, J = 9.1 Hz, 1H, NH–C=O), 7.58–7.49 (m, 4H, C6H4), 4.71 (dd, J = 8.1, 2.4 Hz, 1H, CH–SO2), 3.43 (dt, J = 12.3, 3.3 Hz, 1H, CH–N), 2.74 (s, 3H, CH3), 2.08–1.23 (m, 8H, 4CH2). 13C-NMR (151 MHz, DMSO-d6): δ (ppm) 167.49, 158.84, 139.80, 138.74 (q, 2JFC = 35.79 Hz), 135.08, 131.20, 127.39, 124.01, 121.89, 120.80 (q, 1JFC = 271.35 Hz, CF3), 118.99, 112.60, 60.92, 45.42, 30.51, 24.06, 21.71, 19.44, 18.97. EIMS, m/z 472.5 (M). Elemental analysis for C19H19F3N4O3S2: found C 48.30, H 4.05, N 11.86; calcd C 48.18, H 3.89, N 11.67.

N-(3-fluorophenyl)-2-[1-(3-fluorophenyl)-5-trifluoromethyl-1H-pyrazole-4-carboxylamino]-cyclohexylsulfonamide. III-19. White solid, yield: 70%. mp 167.7~170.5 °C. 1H-NMR (600 MHz, DMSO-d6): δ (ppm) 10.00 (s, 1H, NH-SO2), 8.51 (d, J = 8.9 Hz, 1H, NH–C=O), 8.08 (s, 1H, pyrazole-H), 7.68–6.88 (m, 8H, C6H4 + C6H4), 4.70 (dd, J= 7.2, 3.0 Hz, 1H, CH–SO2), 3.40 (dt, J = 11.4, 3.6 Hz, 1H, CH–N), 2.16–1.24 (m, 8H, 4CH2). 13C-NMR (151 MHz, DMSO-d6): δ (ppm) 162.83 (d, 1JFC = 242.81 Hz, F-Ph), 162.10 (d, 1JFC = 246.73 Hz, F-Ph), 160.07, 140.71, 140.60 (d, 3JFC = 10.87 Hz), 140.28 (d, 3JFC = 10.27 Hz), 131.62 (d, 3JFC = 8.91 Hz), 131.41 (d, 3JFC = 9.66 Hz), 129.97 (d, 2JFC = 39.11 Hz), 122.76, 122.09, 119.64 (q, 1JFC = 270.44 Hz, CF3), 117.52 (d, 2JFC = 20.84 Hz), 115.01, 114.05 (d, 2JFC = 24.46 Hz), 110.30 (d, 2JFC = 20.84 Hz), 106.00 (d, 2JFC = 25.67 Hz), 60.41, 45.10, 30.52, 23.90, 21.92, 19.77. EIMS, m/z 527.10 (M-H+). Elemental analysis for C23H21F5N4O3S: found C 52.27, H 4.01, N 10.60; calcd C 52.03, H 4.21, N 10.43.

N-(2-chlorophenyl)-2-[1-(3-fluorophenyl)-5-trifluoromethyl-1H-pyrazole-4-carboxylamino]-cyclohexylsulfonamide. III-20. White solid, yield: 84%. mp 112.1~115.3 °C. 1H-NMR (600 MHz, DMSO-d6): δ (ppm) 9.38 (s, 1H, NH-SO2), 8.48 (d, J = 9.0 Hz, 1H, NH–C=O), 8.05(s, 1H, pyrazole-H), 7.68–7.23 (m, 8H, C6H4 + C6H4), 4.73 (dd, J= 7.8, 2.4 Hz, 1H, CH–SO2), 3.31(m, 1H, CH–N) (overlap with water peak), 2.15–1.31 (m, 8H, 4CH2). 13C-NMR (151 MHz, DMSO-d6): δ (ppm) 162.09 (d, 1JFC = 246.58 Hz, F-Ph), 160.27, 140.70, 140.25 (d, 3JFC = 10.42 Hz), 134.55, 131.62 (d, 3JFC = 9.21 Hz), 130.24, 129.91 (q, 2JFC = 38.96 Hz), 128.28 (d, 3JFC = 14.50 Hz), 127.51 (d, 3JFC = 8.61 Hz), 122.75, 122.10, 119.62 (q, 1JFC = 270.74 Hz, CF3), 117.59, 117.45, 114.14, 113.98, 62.37, 44.96, 30.77, 24.32, 21.81, 19.62. EIMS, m/z 543.00 (M-H+). Elemental analysis for C23H21ClF4N4O3S: found C 50.69, H 3.88, N 10.28; calcd C 50.56, H 4.03, N 10.42.

N-(4-chlorophenyl)-2-[1-(3-fluorophenyl)-5-trifluoromethyl-1H-pyrazole-4-carboxylamino]-cyclohexylsulfonamide. III-21. White solid, yield: 81%. mp 156.9~158.8 °C. 1H-NMR (600 MHz, DMSO-d6): δ (ppm) 9.88 (s, 1H, NH-SO2), 8.50 (d, J = 8.9 Hz, 1H, NH–C=O), 8.07 (s, 1H, pyrazole-H), 7.68–7.23 (m, 8H, C6H4 + C6H4), 4.69 (dd, J= 7.2, 3.0 Hz, 1H, CH–SO2), 3.32-3.31 (m, 1H, CH–N) (overlap with water peak), 2.16–1.24 (m, 8H, 4CH2). 13C-NMR (151 MHz, DMSO-d6): δ(ppm) 162.10(d, 1JFC = 246.58 Hz, F-Ph), 160.08, 140.74, 140.28 (d, 3JFC = 10.27 Hz), 137.68, 131.61 (d, 3JFC = 9.06 Hz), 129.97 (q, 2JFC = 39.26 Hz), 129.62, 127.95, 122.74, 122.10, 121.22, 119.63 (q, 1JFC = 270.74 Hz, CF3), 117.58, 117.44, 114.13, 113.96, 60.22, 45.02, 30.59, 23.97, 21.84, 19.70. EIMS, m/z 543.00 (M-H+). Elemental analysis for C23H21ClF4N4O3S: found C 50.69, H 3.88, N 10.28; calcd C 50.45, H 4.01, N 10.51.

N-(2-bromophenyl)-2-[1-(3-fluorophenyl)-5-trifluoromethyl-1H-pyrazole-4-carboxylamino]-cyclohexylsulfonamide. III-22. White solid, yield: 92%. mp 101.5~104.8 °C. 1H-NMR (600 MHz, DMSO-d6): δ (ppm) 9.28 (s, 1H, NH-SO2), 8.47 (d, J = 9.0 Hz, 1H, NH–C=O), 8.05 (s, 1H, pyrazole-H), 7.68–7.16 (m, 8H, C6H4 + C6H4), 4.73 (dd, J= 8.4, 3.0 Hz, 1H, CH–SO2), 3.31 (m, 1H, CH–N) (overlap with water peak), 2.11–1.32 (m, 8H, 4CH2). 13C-NMR (151 MHz, DMSO-d6): δ (ppm) 162.10 (d, 1JFC = 246.58 Hz, F-Ph), 160.25, 140.70, 140.26 (d, 3JFC = 10.27 Hz), 133.47, 131.62 (d, 3JFC = 9.06 Hz), 129.91 (q, 2JFC = 39.26 Hz) 128.79, 128.02 (d, 2JFC = 18.72 Hz), 122.75, 122.12, 119.63 (q, 1JFC = 270.74 Hz, CF3), 119.48, 117.58, 117.44, 114.14, 113.97, 62.65, 44.98, 30.81, 24.34, 21.88, 19.61. EIMS, m/z 589.00 (M+H+). Elemental analysis for C23H21BrF4N4O3S: found C 46.87, H 3.59, N 9.51; calcd C 46.98, H 3.33, N 9.72.

N-(3-bromophenyl)-2-[1-(3-fluorophenyl)-5-trifluoromethyl-1H-pyrazole-4-carboxylamino]-cyclohexylsulfonamide. III-23. White solid, yield: 85%. mp 97.7~100.2 °C. 1H-NMR (600 MHz, DMSO-d6): δ (ppm) 9.97 (s, 1H, NH-SO2), 8.51 (d, J = 9.0 Hz, 1H, NH–C=O), 8.07 (s, 1H, pyrazole-H), 7.68–7.23 (m, 8H, C6H4 + C6H4), 4.70 (dd, J= 7.8, 3.0 Hz, 1H, CH–SO2), 3.39 (dt, J = 12.0, 3.6 Hz, 1H, CH–N), 2.18–1.24 (m, 8H, 4CH2). 13C-NMR (151 MHz, DMSO-d6): δ (ppm) 162.10 (d, 1JFC = 246.58 Hz, F-Ph), 160.07, 140.71, 140.28 (d, 3JFC = 10.12 Hz), 131.63 (d, 3JFC = 11.63 Hz), 129.97 (q, 2JFC = 39.11 Hz), 126.50, 122.75, 122.40, 122.08, 121.60, 119.63 (q, 1JFC = 270.74 Hz, CF3), 118.07, 117.59, 117.45, 114.13, 113.97, 60.60, 45.06, 30.54, 23.91, 21.92, 19.73. EIMS, m/z 589.00 (M+H+). Elemental analysis for C23H21BrF4N4O3S: found C 46.87, H 3.59, N 9.51; calcd C 46.59, H 3.74, N 9.32.

N-(4-bromophenyl)-2-[1-(3-fluorophenyl)-5-trifluoromethyl-1H-pyrazole-4-carboxylamino]-cyclohexylsulfonamide. III-24. White solid, yield: 91%. mp 169.7~171.2 °C. 1H-NMR (600 MHz, DMSO-d6): δ (ppm) 10.00 (s, 1H, NH-SO2), 8.51 (d, J = 9.0 Hz, 1H, NH–C=O), 8.08(s, 1H, pyrazole-H), 7.68–6.88 (m, 8H, C6H4 + C6H4), 4.70 (dd, J= 7.2, 3.0 Hz, 1H, CH–SO2), 3.40 (dt, J = 11.4, 3.6 Hz, 1H, CH–N), 2.18-1.24 (m, 8H, 4CH2). 13C-NMR (151 MHz, DMSO-d6): δ (ppm) 162.10 (d, 1JFC = 246.58 Hz, F-Ph), 160.08, 140.74, 140.28 (d, 3JFC = 10.12 Hz), 138.14, 132.52, 131.61 (d, 3JFC = 9.06 Hz), 129.97 (q, 2JFC = 38.96 Hz), 122.74, 122.09, 121.51, 119.64 (q, 1JFC = 270.74 Hz, CF3), 117.58, 117.44, 115.94, 114.13, 113.97, 60.23, 45.01, 30.59, 23.97, 21.84, 19.69. EIMS, m/z 589.00 (M+H+). Elemental analysis for C23H21BrF4N4O3S: found C 46.87, H 3.59, N 9.51; calcd C 46.65, H 3.48, N 9.29.

N-(2,4,5-trifluorophenyl)-2-[1-(3-fluorophenyl)-5-trifluoromethyl-1H-pyrazole-4-carboxylamino]-cyclohexylsulfonamide. III-25. White solid, yield: 70%. mp 100.0~121.1 °C. 1H-NMR (600 MHz, DMSO-d6): δ (ppm) 9.81 (s, 1H, NH-SO2), 8.47 (s, 1H, NH–C=O), 8.06(s, 1H, pyrazole-H), 7.66–7.39 (m, 6H, C6H4 + C6H2), 4.69 (s, 1H, CH–SO2), 3.41 (s, 1H, CH–N), 2.14–1.32 (m, 8H, 4CH2). 13C-NMR (151 MHz, DMSO-d6): δ (ppm) 162.10 (d, 1JFC = 246.43 Hz, F-Ph), 160.14, 151.17 (dd, JFC = 245.38, 9.66 Hz, F-Ph), 146.92 (dt, JFC = 244.77, 11.17 Hz, F-Ph), 145.99 (dd, JFC = 239.03, 12.68 Hz, F-Ph), 140.68, 140.27 (d, 3JFC = 10.42 Hz), 131.61 (d, 3JFC = 9.06 Hz), 129.96 (q, 2JFC = 39.26 Hz), 122.73, 122.02, 119.61 (q, 1JFC = 270.74 Hz, CF3), 117.51 (d, 2JFC = 21.14 Hz), 114.75 (d, 2JFC = 21.14 Hz), 114.03 (d, 2JFC = 25.67 Hz), 106.76 (d, 2JFC = 22.20 Hz), 106.58(d, 2JFC = 22.05 Hz), 61.60, 44.91, 30.60, 24.13, 21.68, 19.63. EIMS, m/z 563.00 (M-H+). Elemental analysis for C23H21F7N4O3S: found C 48.94, H 3.39, N 9.93; calcd C 48.76, H 3.46, N 9.71.

N-(3-chlorophenyl)-2-[1-(3-fluorophenyl)-5-trifluoromethyl-1H-pyrazole-4-carboxylamino]-cyclohexylsulfonamide. III-26. White solid, yield: 83%. mp 137.2~140.8 °C. 1H-NMR (300 MHz, DMSO-d6): δ (ppm) 10.07 (s, 1H, NH-SO2), 8.50 (d, J = 9.0 Hz, 1H, NH–C=O), 8.05(s, 1H, pyrazole-H), 7.86–7.38 (m, 8H, C6H4 + C6H4), 4.73 (dd, J= 8.7, 3.0 Hz, 1H, CH–SO2), 3.45–3.38 (m, 1H, CH–N) (overlap with water peak), 2.16–1.36 (m, 8H, 4CH2). 13C-NMR (151 MHz, DMSO-d6): δ (ppm) 162.09 (d, 1JFC = 246.58 Hz, F-Ph), 160.09, 140.67, 140.27 (d, 3JFC = 10.12 Hz), 139.62, 134.56, 134.23, 131.61 (d, 3JFC = 9.21 Hz), 129.93 (q, 2JFC = 38.81 Hz), 127.21, 127.00, 122.75, 122.10, 119.62 (q, 1JFC = 270.59 Hz, CF3), 117.51 (d, 2JFC = 20.84 Hz), 114.06 (d, 2JFC = 24.61 Hz), 109.43, 62.19, 44.93, 30.74, 24.21, 21.87, 19.63. EIMS, m/z 544.10 (M). Elemental analysis for C23H21ClF4N4O3S: found C 50.69, H 3.88, N 10.28; calcd C 50.77, H 3.74, N 10.39.

N-(2-fluorophenyl)-2-[1-(3-fluorophenyl)-5-trifluoromethyl-1H-pyrazole-4-carboxylamino]-cyclohexylsulfonamide. III-27. White solid, yield: 77%. mp 128.3~131.5 °C. 1H-NMR (600 MHz, DMSO-d6): δ (ppm) 9.56 (s, 1H, NH-SO2), 8.47 (d, J = 8.4 Hz, 1H, NH–C=O), 8.05(s, 1H, pyrazole-H), 7.68–7.17 (m, 8H, C6H4 + C6H4), 4.70 (dd, J= 8.4, 3.6 Hz, 1H, CH–SO2), 3.29 (dt, J = 12.0, 3.6 Hz, 1H, CH–N), 2.16–1.27 (m, 8H, 4CH2). 13C-NMR (151 MHz, DMSO-d6): δ (ppm) 162.10 (d, 1JFC = 246.43 Hz, F-Ph), 160.23, 155.68 (d, 1JFC = 245.68 Hz, F-Ph), 140.72, 140.27 (d, 3JFC = 10.27 Hz), 131.61 (d, 3JFC = 9.06 Hz), 129.92 (d, 2JFC = 39.11 Hz), 127.26 (d, 3JFC = 7.40 Hz), 126.85, 125.29 (d, 3JFC = 12.68 Hz), 125.14, 122.74, 122.11, 119.63 (q, 1JFC = 270.74 Hz, CF3), 117.51 (d, 2JFC = 21.14 Hz), 116.42 (d, 2JFC = 19.93 Hz), 114.05 (d, 2JFC = 24.61 Hz), 61.36, 44.98, 30.67, 24.19, 21.75, 19.68. EIMS, m/z 527.10 (M-H+). Elemental analysis for C23H21F5N4O3S: found C 52.27, H 4.01, N 10.60; calcd C 52.45, H 3.88, N 10.81.

N-(3-cyanophenyl)-2-(1-methyl-3-difluoromethyl-1H-pyrazole-4-carboxylamino]-cyclohexylsulfonamide III-28. White solid, yield: 71%. mp 213.8~215.2 °C. 1H-NMR (600 MHz, DMSO-d6):δ (ppm) 10.13 (s, 1H, NH-SO2), 8.36 (s, 1H, pyrazole-H), 7.67(d, J = 8.4 Hz, 1H, NH–C=O), 7.52–7.44 (m, 4H, C6H4), 7.26 (t, J = 54 Hz, 1H, HCF2), 4.64 (dd, J = 7.2, 3.0 Hz, 1H, CH–SO2), 3.93 (s, 3H, CH3), 3.52 (dt, J = 11.4, 3.6 Hz, 1H, CH–N), 2.16–1.29 (m, 8H, 4CH2). 13C-NMR (151 MHz, DMSO-d6): δ (ppm) 161.06, 144.78 (t, 2JFC = 22.80 Hz), 139.71, 133.46, 130.96, 127.06, 123.73, 121.63, 118.87, 116.34, 112.38, 110.16 (t, 1JFC = 234.05 Hz, HCF2), 61.11, 44.92, 40.40, 30.38, 23.84, 22.06, 19.91. EIMS, m/z 436.10 (M-H+). Elemental analysis for C19H21F2N5O3S: found C 52.17, H 4.84, N 16.01; calcd C 52.37, H 4.69, N 15.83.

N-(2-fluorophenyl)-2-(1-methyl-3-difluoromethyl-1H-pyrazole-4-carboxylamino]-cyclohexylsulfonamide III-29. White solid, yield: 65%. mp 164.7~165.9 °C. 1H-NMR (600 MHz, DMSO-d6): δ (ppm) 9.55 (s, 1H, NH-SO2), 8.38 (s, 1H, pyrazole-H), 7.67 (d, J = 8.4 Hz, 1H, NH–C=O), 7.42–7.13 (m, 4H, C6H4), 7.33(t, J = 53.4 Hz, 1H, HCF2), 4.65 (dd, J = 7.8, 3.6 Hz, 1H, CH–SO2), 3.93 (s, 3H, CH3), 3.30 (dt, J = 12.0, 3.6 Hz, 1H, CH–N), 2.13–1.30 (m, 8H, 4CH2). 13C-NMR (151 MHz, DMSO-d6): δ (ppm) 161.24, 155.57 (d, 1JFC = 245.68 Hz, F-Ph), 144.67 (t, 2JFC = 23.41 Hz), 133.56, 127.17 (d, 3JFC = 7.55 Hz), 126.66, 125.27 (d, 3JFC = 12.68 Hz), 125.10 (d, 3JFC = 3.47 Hz), 116.43, 116.30, 110.17 (t, 1JFC = 234.20 Hz, HCF2), 61.32, 44.87, 40.41, 30.49, 24.05, 21.93, 19.89. EIMS, m/z 429.10 (M-H+). Elemental analysis for C18H21F3N4O3S: found C 50.23, H 4.92, N 13.02; calcd C 49.98, H 5.21, N 13.19.

N-(2-bromophenyl)-2-(1-methyl-3-difluoromethyl-1H-pyrazole-4-carboxylamino]-cyclohexylsulfonamide III-30. White solid, yield: 90%. mp 183.3~184.1°C. 1H-NMR (600 MHz, DMSO-d6): δ (ppm) 9.25 (s, 1H, NH-SO2), 8.39 (s, 1H, pyrazole-H), 7.69 (d, J = 8.4 Hz, 1H, NH–C=O), 7.66–7.13 (m, 4H, C6H4), 7.25(t, J = 54.6 Hz, 1H, HCF2), 4.69 (dd, J = 7.8, 3.6 Hz, 1H, CH–SO2), 3.93 (s, 3H, CH3), 3.37 (dt, J = 11.4, 3.6 Hz, 1H, CH–N), 2.09–1.36 (m, 8H, 4CH2). 13C-NMR (151 MHz, DMSO-d6): δ (ppm) 161.28, 144.69 (t, 2JFC = 23.25 Hz), 135.92, 133.56, 133.44, 128.75, 127.84, 127.77, 119.27, 116.41, 110.17 (t, 1JFC = 234.20 Hz, HCF2), 62.56, 44.86, 40.81, 30.62, 24.20, 22.04, 19.83. EIMS, m/z 491.00 (M+H+). Elemental analysis for C18H21BrF2N4O3S: found C 44.00, H 4.31, N 11.40; calcd C 44.21, H 4.19, N 11.56.

N-(2-chlorophenyl)-2-(1-methyl-3-difluoromethyl-1H-pyrazole-4-carboxylamino]-cyclohexylsulfonamide. III-31. White solid, yield: 67%. mp 190.8~192.3 °C. 1H-NMR (300 MHz, DMSO-d6): δ(ppm) 9.37 (s, 1H, NH-SO2), 8.39 (s, 1H, pyrazole-H), 7.70 (d, J = 8.4 Hz, 1H, NH–C=O), 7.51–7.19 (m, 4H, C6H4), 7.24(t, J = 54.3 Hz, 1H, HCF2), 4.68 (dd, J = 7.5, 3.3 Hz, 1H, CH–SO2), 3.94 (s, 3H, CH3), 3.33–3.31 (m, 1H, CH–N) (overlap with water peak), 2.16–1.34 (m, 8H, 4CH2). 13C-NMR (151 MHz, DMSO-d6): δ (ppm) 161.29, 144.68 (t, 2JFC = 23.56 Hz), 134.54, 133.57, 130.20, 128.17, 128.15, 127.35, 127.26, 116.41, 110.17 (t, 1JFC = 234.20 Hz, HCF2), 62.31, 44.86, 40.42, 30.58, 24.17, 21.99, 19.84. EIMS, m/z 445.10 (M-H+). Elemental analysis for C18H21ClF2N4O3S: found C 48.38, H 4.74, N 12.54; calcd C 48.21, H 4.92, N 12.34.

N-(3-fluorophenyl)-2-(1-methyl-3-difluoromethyl-1H-pyrazole-4-carboxylamino]-cyclohexylsulfonamide. III-32. White solid, yield: 84%. mp 185.7~187.1 °C. 1H-NMR (600 MHz, DMSO-d6): δ (ppm) 9.97 (s, 1H, NH-SO2), 8.38 (s, 1H, pyrazole-H), 7.70 (d, J = 8.4 Hz, 1H, NH–C=O), 7.33–6.84 (m, 4H, C6H4), 7.27(t, J = 54.6 Hz, 1H, HCF2), 4.62 (dd, J= 7.2, 3.6 Hz, 1H, CH–SO2), 3.94 (s, 3H, CH3), 3.43 (dt, J = 11.4, 3.6 Hz, 1H, CH–N), 2.13–1.26 (m, 8H, 4CH2). 13C-NMR (151 MHz, DMSO-d6): δ (ppm) 162.79 (d, 1JFC = 242.81 Hz, F-Ph), 161.11, 144.76 (t, 2JFC = 23.25 Hz), 140.57 (d, 3JFC = 10.72 Hz), 133.51, 131.29 (d, 3JFC = 9.66 Hz), 116.42, 114.95, 110.18 (t, 1JFC = 234.50 Hz, HCF2), 106.01, 105.84, 60.48, 45.02, 40.42, 30.36, 23.78, 22.11, 20.01. EIMS, m/z 429.10 (M-H+). Elemental analysis for C18H21F3N4O3S: found C 50.23, H 4.92, N 13.02; calcd C 50.44, H 5.16, N 12.89.

N-(4-bromophenyl)-2-(1-methyl-3-difluoromethyl-1H-pyrazole-4-carboxylamino]-cyclohexylsulfonamide III-33. White solid, yield: 84%. mp 229.9~231.1 °C. 1H-NMR (600 MHz, DMSO-d6): δ (ppm) 9.86 (s, 1H, NH-SO2), 8.37 (s, 1H, pyrazole-H), 7.69 (d, J = 8.4 Hz, 1H, NH–C=O), 7.47–7.14 (m, 4H, C6H4), 7.27 (t, J = 54.0 Hz, 1H, HCF2), 4.61 (dd, J= 7.8, 3.6 Hz, 1H, CH–SO2), 3.94 (s, 3H, CH3), 3.34 (m, 1H, CH–N) (overlap with water peak), 2.11–1.23 (m, 8H, 4CH2). 13C-NMR (151 MHz, DMSO-d6): δ (ppm) 161.12, 144.71 (t, 2JFC = 23.25 Hz), 138.11, 133.53, 132.42 (two carbon atoms), 121.41 (two carbon atoms), 116.42, 115.85, 110.20 (t, 1JFC = 234.35 Hz, HCF2), 60.30, 44.94, 40.40, 30.40, 23.83, 22.03, 19.93. EIMS, m/z 491.00 (M+H+). Elemental analysis for C18H21BrF2N4O3S: found C 44.00, H 4.31, N 11.40; calcd C 43.87, H 4.19, N 11.56.

N-(3-bromophenyl)-2-(1-methyl-3-difluoromethyl-1H-pyrazole-4-carboxylamino]-cyclohexylsulfonamide. III-34. White solid, yield: 79%. mp 221.1~223.8 °C. 1H-NMR (600 MHz, DMSO-d6): δ (ppm) 9.93 (s, 1H, NH-SO2), 8.37 (s, 1H, pyrazole-H), 7.69(d, J = 8.4 Hz, 1H, NH–C=O), 7.35–7.18 (m, 4H, C6H4), 7.27(t, J = 54.0 Hz, 1H, HCF2), 4.62 (dd, J= 7.2, 3.6 Hz, 1H, CH–SO2), 3.94 (s, 3H, CH3), 3.42 (dt, J = 11.4, 3.6 Hz, 1H, CH–N), 2.13–1.29 (m, 8H, 4CH2). 13C-NMR (151 MHz, DMSO-d6): δ (ppm) 161.10, 144.87 (t, 2JFC = 23.41 Hz), 140.43, 133.48, 131.53, 126.35, 122.33, 121.51, 117.97, 116.40, 110.18 (t, 1JFC = 234.05 Hz, HCF2), 60.74, 45.00, 40.43, 30.37, 23.78, 22.13, 19.99. EIMS, m/z 491.00 (M+H+). Elemental analysis for C18H21BrF2N4O3S: found C 44.00, H 4.31, N 11.40; calcd C 44.19, H 4.52, N 11.27.

N-(2,4,5-trifluorophenyl)-2-(1-methyl-3-difluoromethyl-1H-pyrazole-4-carboxylamino]-cyclohexylsulfonamide III-35. White solid, yield: 89%. mp 68.5~70.4 °C. 1H-NMR (600 MHz, DMSO-d6): δ (ppm) 9.78 (s, 1H, NH-SO2), 8.37 (s, 1H, pyrazole-H), 7.66 (d, J = 8.4 Hz, 1H, NH–C=O), 7.64–7.43 (m, 2H, C6H2), 7.23(t, J = 54.6 Hz, 1H, HCF2), 4.63 (dd, J= 7.2, 3.6 Hz, 1H, CH–SO2), 3.93 (s, 3H, CH3), 3.42 (dt, J = 11.4, 3.6 Hz, 1H, CH–N), 2.11–1.33 (m, 8H, 4CH2). 13C-NMR (151 MHz, DMSO-d6): δ (ppm) 161.17, 150.94 (dd, 1JFC = 245.38, 8.91 Hz, F-Ph), 146.78 (dt, JFC = 244.47, 14.19 Hz, F-Ph), 145.95 (dd, 1JFC = 242.36, 12.84 Hz, F-Ph) 144.70 (t, 2JFC = 23.25 Hz), 133.51, 122.14, 116.30, 114.40 (d, 2JFC = 22.20 Hz), 110.13 (t, 1JFC = 234.35 Hz, HCF2), 106.57 (dd, JFC = 26.58, 22.05 Hz), 61.73, 44.78, 40.41, 30.45, 24.02, 21.85, 19.80. EIMS, m/z 466.11 (M). Elemental analysis for C18H21F5N4O3S: found C 46.35, H 4.11, N 12.01; calcd C 46.52, H 3.98, N 12.24.

N-(4-chlorophenyl)-2-(1-methyl-3-difluoromethyl-1H-pyrazole-4-carboxylamino]-cyclohexylsulfonamide. III-36. White solid, yield: 55%. mp 255.7~257.0 °C. 1H-NMR (600 MHz, DMSO-d6): δ (ppm) 9.84 (s, 1H, NH-SO2), 8.37 (s, 1H, pyrazole-H), 7.68 (d, J = 8.4 Hz, 1H, NH–C=O), 7.34–7.19 (m, 4H, C6H4), 7.27(t, J = 54.0 Hz, 1H, HCF2), 4.61 (dd, J= 6.6, 3.0 Hz, 1H, CH–SO2), 3.94 (s, 3H, CH3), 3.35 (dt, J = 11.4, 3.0 Hz, 1H, CH–N), 2.14–1.25 (m, 8H, 4CH2). 13C-NMR (151 MHz, DMSO-d6): δ (ppm) 161.11, 144.72 (t, 2JFC = 23.57 Hz), 137.67, 133.52, 129.53 (two carbon atoms), 127.86, 121.09 (two carbon atoms), 116.43, 110.20 (t, 1JFC = 234.50 Hz, HCF2), 60.29, 44.94, 40.44, 30.41, 23.84, 22.04, 19.94. EIMS, m/z 447.10 (M+H+). Elemental analysis for C18H21ClF2N4O3S: found C 48.38, H 4.74, N 12.54; calcd C 48.54, H 4.51, N 12.70.

3.3. Fungicidal Activity Bioassays

The in vitro and in vivo fungicidal activities of all the target compounds against B. cinerea were tested by mycelium growth inhibition assay, spore germination experiment and tomato pot experiment, respectively. The B. cinerea strains (KZ-9, CY-09) were collected from different regions of Liaoning, China, and cultured on potato dextrose agar (PDA) for several generations. Boscalid and procymidone (with the purity more than 95%) are provided by the Shenyang Research Institute of the Chemical Industry.

3.3.1. Evaluation of the Fungicidal Activity on B. cinerea by Mycelium Growth Experiments

Using the mycelium growth rate method, the compounds were dissolved in acetone and mixed with sterile molten PDA to obtain four concentrations of 50, 12.50, 3.125 and 0.78 μg/mL, poured them into sterile 90 mm diameter Petri dishes under aseptic conditions, respectively. A 5 mm B. cinerea plug was inoculated in the center of 90 mm PDA petri dish. Boscalid and procymidone were used as positive control agents while acetone was used as blank control. The EC50 value was calculated according to mycelium growth. [25]

3.3.2. Evaluation of the Fungicidal Activity on B. cinerea by Spore Germination Experiments

The effect of target compounds III on spore germination against B. cinerea was determined by the concave slide method. The method was given in reference [25]. The commercial fungicide boscalid was used as the positive control, and the results were given in Table 3.

3.3.3. In Vivo Fungicidal Activity against B. cinerea by Tomato Pot Experiments

The compound was first prepared to 5% emulsifiable concentrate (EC) and diluted with water into an aqueous solution of 200 μg/mL. Tomato seedlings at the 6–8 leaves stage on the main stem were selected as objects. Spraying the solution evenly on the tomato plants and upon naturally drying, a spore suspension of B. cinerea with 5 × 105 spores per mL was evenly sprayed on the tomato seedlings. It was placed in a greenhouse at 20–25 °C, with humidity above 80%. Waiting for observation, using boscalid as control agents, and an emulsifiable concentrate without agent was the blank control and then disease incidence and index on tomato leaves were calculated. [25]

4. Conclusions

In conclusion, 36 novel sulfonamide compounds were designed and synthesized in this paper. The thiazole and pyrazole active groups were introduced by the method of active substructure splicing, and benzene ring (linked to NH) was optimized. In vitro and in vivo fungicidal activities against B.cinerea were evaluated, and some target compounds showed excellent activity. Moreover, the structure–activity relationship revealed that the compounds containing phenylpyrazole amide structure showed better activity effects, while fungicidal activity was highest at the meta-substituted fluorine atom on the benzene ring (linked to NH), such as compound III-19 showing notable in vitro fungicidal activity. Thus, the present results laid the foundation for further structural design and fungicide screening.