药学学报

Compd.	Yield/%	mp/℃		Compd.	Yield/%	mp/℃
2a	55	160.8-161.8		2m	62	148.1-149.1
2b	45	157.3-158.3	2n	58	111.7-112.7
2c	41	172.1-172.1	2o	55	106.1-107.1
2d	53	174.2-175.2	2p	44	138.9-139.9
2e	40	174.7-175.7	2q	39	141.3-142.3
2f	49	176.3-177.3	2r	43	124.8-125.8
2g	58	199.6-200.6	2s	56	141.6-142.6
2h	53	142.2-143.2	2t	62	115.0-116.0
2i	49	159.8-160.8	2u	54	117.7-118.7
2j	40	197.1-198.1	2v	64	136.2-137.2
2k	48	203.7-204.7	2w	61	139.3-149.3
2l	52	162.5-163.5	2x	61	131.5-132.5

Compd.	Yield/%	mp/℃		Compd.	Yield/%	mp/℃
2a	55	160.8-161.8		2m	62	148.1-149.1
2b	45	157.3-158.3	2n	58	111.7-112.7
2c	41	172.1-172.1	2o	55	106.1-107.1
2d	53	174.2-175.2	2p	44	138.9-139.9
2e	40	174.7-175.7	2q	39	141.3-142.3
2f	49	176.3-177.3	2r	43	124.8-125.8
2g	58	199.6-200.6	2s	56	141.6-142.6
2h	53	142.2-143.2	2t	62	115.0-116.0
2i	49	159.8-160.8	2u	54	117.7-118.7
2j	40	197.1-198.1	2v	64	136.2-137.2
2k	48	203.7-204.7	2w	61	139.3-149.3
2l	52	162.5-163.5	2x	61	131.5-132.5

Compd.	¹H NMR, ¹³C NMR, HRMS-ESI
2a	¹H NMR (400 MHz, DMSO-d₆) δ: 8.02 (d, J = 8.5 Hz, 2H, ArH), 7.86 (d, J = 9.2 Hz, 2H, ArH), 4.60 (s, 2H, CH₂-S), 3.61 (d, J = 2.9 Hz, 2H, NH₂CH₂), 2.09 (d, J = 6.7 Hz, 2H, CH₂-cyclohexyl), 1.79 (s, 3H, Me), 1.66 (m, 7H, Cyclohexyl), 1.54 (m, 4H, cyclohexyl); ¹³C NMR (100 MHz, DMSO-d₆) δ: 190.3, 174.3, 161.5 (2C), 155.9, 142.8, 131.1, 130.0 (2C), 118.7 (2C), 115.3, 43.9, 37.6 (2C), 36.8 (2C), 32.9, 32.6 (3C), 10.9; HRMS-ESI: m/z calcd. for C₂₂H₂₈N₄O₃S [M+H]⁺ 429.194 6, found 429.194 8.
2b	¹H NMR (400 MHz, DMSO-d₆) δ: 8.03 (d, J = 8.5 Hz, 2H, ArH), 7.83 (d, J = 9.9 Hz, 2H, ArH), 4.60 (s, 2H, CH₂-S), 3.60 (d, J = 2.9 Hz, 2H, NH₂CH₂), 2.29-2.26 (m, 2H, CH₂CH₃), 2.08 (d, J = 6.9 Hz, 2H, CH₂-cyclohexyl), 1.65 (m, 7H, cyclohexyl), 1.53 (dd, J = 6.1, 2.1 Hz, 4H, cyclohexyl), 0.91 (s, 3H, CH₂CH₃); ¹³C NMR (100 MHz, DMSO-d₆) δ: 192.3, 174.4, 161.7 (2C), 156.4, 143.2, 131.0, 130.0 (2C), 118.7 (2C), 117.2, 43.9, 37.6, 36.6 (2C), 32.9 (2C), 32.6 (3C), 22.5, 13.7; HRMS-ESI: m/z calcd. for C₂₃H₃₀N₄O₃S [M+H]⁺ 443.210 3, found 443.210 5.
2c	¹H NMR (400 MHz, DMSO-d₆) δ: 8.02 (s, 2H, ArH), 7.88-7.79 (m, 2H, ArH), 4.63-4.55 (m, 2H, CH₂-S), 3.62-3.55 (m, 1H, CH-NH₂), 2.15-2.04 (m, 2H, CH₂-cyclohexyl), 1.83-1.77 (m, 3H, Me), 1.68-1.60 (m, 1H, cyclohexyl), 1.54-1.48 (m, 3H, CHCH₃), 1.35-1.21 (m, 7H, cyclohexyl), 1.02-0.90 (m, 3H, cyclohexyl); ¹³C NMR (100 MHz, DMSO-d₆) δ: 191.9, 174.7, 164.1, 160.9, 156.4, 143.4, 131.0, 129.5 (2C), 118.3 (2C), 114.7, 50.87, 36.6, 36.3, 32.4 (2C), 25.8, 25.5 (3C), 20.7, 10.4; HRMS-ESI: m/z calcd. for C₂₃H₃₀N₄O₃S [M+H]⁺ 443.210 3, found 443.210 6.
2d	¹H NMR (400 MHz, DMSO-d₆) δ: 8.02 (d, J = 8.4 Hz, 2H, ArH), 7.82 (d, J = 8.5 Hz, 2H, ArH), 4.60 (s, 2H, CH₂-S), 3.54 (q, J = 7.1 Hz, 1H, CH-NH₂), 2.28 (q, J = 7.4 Hz, 2H, CH₂CH₃), 2.08 (d, J = 6.7 Hz, 2H, CH₂-cyclohexyl), 1.50-1.40 (m, 3H, CHCH₃), 1.36-1.28 (m, 2H, cyclohexyl), 1.25 (t, J = 6.1 Hz, 5H, cyclohexyl), 0.91 (t, J = 7.2 Hz, 7H, cyclohexyl and CH₂CH₃); ¹³C NMR (100 MHz, DMSO-d₆) δ: 191.9, 174.9, 163.2, 160.44, 156.32, 143.51, 130.96, 129.55 (2C), 118.33 (2C), 114.63, 50.95, 36.76, 36.14, 32.50 (2C), 25.83, 25.55 (3C), 20.87, 17.97, 13.27; HRMS-ESI: m/z calcd. for C₂₄H₃₂N₄O₃S [M+H]⁺ 457.225 9, found 457.226 1.
2e	¹H NMR (400 MHz, DMSO-d₆) δ: 8.07 (s, 2H, ArH), 7.86-7.72 (m, 2H, ArH), 4.60 (s, 2H, CH₂-S), 3.72 (m, 1H, CH-NH₂), 2.15-2.01 (m, 2H, CH₂-cyclohexyl), 1.80-1.74 (s, 3H, Me), 1.71-1.62 (m, 1H, cyclohexyl), 1.52-1.44 (m, 3H, CHCH₃), 1.36-1.09 (m, 7H, cyclohexyl), 1.03-0.90 (m, 3H, cyclohexyl); ¹³C NMR (100 MHz, DMSO-d₆) δ: 191.6, 174.7, 163.2, 160.5, 154.1, 144.0, 131.8, 129.0 (2C), 117.8 (2C), 114.0, 51.0, 36.8, 36.3, 32.8 (2C), 25.9, 25.3 (3C), 18.8, 10.3; HRMS-ESI: m/z calcd. for C₂₃H₃₀N₄O₃S [M+H]⁺ 443.210 3, found 443.210 7.
2f	¹H NMR (400 MHz, DMSO-d₆) δ: 8.04 (d, J = 8.4 Hz, 2H, ArH), 7.80 (d, J = 8.5 Hz, 2H, ArH), 4.61 (s, 2H, CH₂-S), 3.60 (q, J = 7.8 Hz, 1H, CH-NH₂), 2.18 (q, J = 7.2 Hz, 2H, CH₂CH₃), 2.08 (d, J = 7.0 Hz, 2H, CH₂-cyclohexyl), 1.56-1.40 (m, 3H, CHCH₃), 1.35-1.26 (m, 2H, cyclohexyl), 1.21 (t, J = 6.1 Hz, 5H, cyclohexyl), 0.91 (t, J = 7.2 Hz, 7H, cyclohexyl and CH₂CH₃); ¹³C NMR (100 MHz, DMSO-d₆) δ: 191.2, 174.6, 163.8, 161.1, 156.3, 144.7, 131.0, 129.5 (2C), 118.1 (2C), 114.3, 51.8, 36.9 (2C), 35.4 (2C), 31.6, 26.6 (2C), 25.1, 20.2, 18.3, 14.1; HRMS-ESI: m/z calcd. for C₂₄H₃₂N₄O₃S [M+H]⁺ 457.225 9, found 457.226 3.
2g	¹H NMR (400 MHz, DMSO-d₆) δ: 8.09 (d, J = 8.4 Hz, 2H, ArH), 7.82 (d, J = 8.5 Hz, 2H, ArH), 4.68 (s, 2H, CH₂-S), 3.11 (m, 2H, NH₂CH₂CH₂), 2.60 (m, 2H, NH₂CH₂CH₂), 2.28 (m, 2H, CH₂CH₃), 2.08 (d, J = 7.8 Hz, 2H, CH₂-cyclohexyl), 1.56-1.29 (m, 11H, cyclohexyl), 0.91 (t, J = 7.2 Hz, 3H, CH₂CH₃); ¹³C NMR (100 MHz, DMSO-d₆) δ: 191.6, 173.6, 162.1, 160.8, 157.9, 143.6, 131.0, 129.6 (2C), 119.2 (2C), 114.3, 39.8, 36.9, 36.0, 34.5, 33.8 (2C), 32.7, 26.4, 25.3 (2C), 20.2, 14.2; HRMS-ESI: m/z calcd. for C₂₄H₃₂N₄O₃S [M+H]⁺ 457.225 9, found 457.226 0.
2h	¹H NMR (400 MHz, DMSO-d₆) δ: 8.02 (d, J = 8.5 Hz, 2H, ArH), 7.82 (d, J = 8.8 Hz, 2H, ArH), 4.58 (s, 2H, CH₂-S), 3.40 (t, J = 6.6 Hz, 1H, CH-NH₂), 2.07 (s, 3H, Me), 1.79 (m, 4H, CH₂-cyclohexyl and CH₂CH₂CH₂CH₃), 1.66-0.91 (m, 15H, cyclohexyl and CH₂CH₂CH₂CH₃), 0.84 (t, J = 6.9 Hz, 3H, CH₂CH₂CH₂CH₃); ¹³C NMR (100 MHz, DMSO-d₆) δ: 192.0, 174.7, 163.7, 161.0, 156.2, 143.5, 131.0, 129.5 (2C), 118.3 (2C), 114.9, 55.4, 36.6, 36.2, 34.4, 32.4 (2C), 27.3, 25.8 (3C), 25.5, 22.1, 13.9, 10.4; HRMS-ESI: m/z calcd. for C₂₆H₃₆N₄O₃S [M+H]⁺ 485.257 2, found 485.256 8.
2i	¹H NMR (400 MHz, DMSO-d₆) δ: 8.06 (d, J = 8.4 Hz, 2H, ArH), 7.84 (d, J = 8.7 Hz, 2H, ArH), 4.59 (s, 2H, CH₂-S), 3.37 (t, J = 6.5 Hz, 1H, CH-NH₂), 2.28 (q, J = 7.3 Hz, 2H, CH₂CH₃), 2.06 (d, J = 6.7 Hz, 2H, CH₂-cyclohexyl), 1.69-1.17 (m, 13H, cyclohexyl and CH₂CH₂CH₂CH₃), 0.88 (dt, J = 25.0, 7.2 Hz, 10H, CH₂CH₃ and CH₂CH₂CH₂CH₃); ¹³C NMR (100 MHz, DMSO-d₆) δ: 191.9, 174.8, 163.1 (2C), 156.2, 143.5, 130.9, 129.5 (2C), 120.9 (2C), 118.3, 55.5, 36.7, 36.1, 34.5, 32.4 (2C), 27.4, 25.8, 25.5 (2C), 22.1, 17.9, 13.8, 13.2; HRMS-ESI: m/z calcd. for C₂₇H₃₈N₄O₃S [M+H]⁺ 499.272 9, found 499.272 6.
2j	¹H NMR (400 MHz, DMSO-d₆) δ: 8.02 (d, J = 8.4 Hz, 2H, ArH), 7.82 (d, J = 8.8 Hz, 2H, ArH), 4.63 (s, 1H, CH₂-S), 2.78 (t, J = 7.6 Hz, 2H, NH₂CH₂CH₂CH₂CH₂CH₂), 2.40 (t, J = 7.3 Hz, 2H, NH₂CH₂CH₂CH₂CH₂CH₂), 2.28 (d, J = 7.6 Hz, 2H, CH₂-cyclohexyl), 2.08 (s, 3H, Me), 1.66 (q, J = 5.8 Hz, 4H, NH₂CH₂CH₂CH₂CH₂CH₂), 1.61-1.57 (m, 4H, cyclohexyl and NH₂CH₂CH₂CH₂CH₂CH₂), 1.57-1.50 (m, 4H, cyclohexyl), 1.37-1.25 (m, 5H, cyclohexyl); ¹³C NMR (100 MHz, DMSO-d₆) δ: 192.1, 175.5, 164.1 (2C), 155.3, 144.2, 131.0, 129.8 (2C), 118.0 (2C), 117.0, 43, 0, 37.1, 32.9 (2C), 27.2, 26.4 (2C), 25.7 (3C), 24.6, 13.7; HRMS-ESI: m/z calcd. for C₂₆H₃₆N₄O₃S [M+H]⁺ 485.257 2, found 485.257 7.
2k	¹H NMR (400 MHz, DMSO-d₆) δ: 8.00 (d, J = 8.5 Hz, 2H, ArH), 7.82 (d, J = 8.6 Hz, 2H, ArH), 4.60 (s, 2H, CH₂-S), 2.76 (t, J = 7.6 Hz, 2H, NH₂CH₂CH₂CH₂CH₂CH₂), 2.40 (t, J = 7.3 Hz, 2H, NH₂CH₂CH₂CH₂CH₂CH₂), 2.30 (m, 2H, CH₂CH₃), 2.08 (d, J = 6.8 Hz, 2H, CH₂-cyclohexyl), 1.66 (q, J = 5.8 Hz, 4H, NH₂CH₂CH₂CH₂CH₂CH₂), 1.61-1.57 (m, 4H, cyclohexyl), 1.57-1.50 (m, 4H, cyclohexyl and NH₂CH₂CH₂CH₂CH₂CH₂), 1.37-1.25 (m, 5H, cyclohexyl), 0.92 (m, 3H, CH₂CH₃); ¹³C NMR (100 MHz, DMSO-d₆) δ: 192.3, 175.8, 164.3 (2C), 155.2, 144.5, 131.1, 129.9 (2C), 118.6 (2C), 117.2, 42.8, 37.2 (2C), 32.9 (2C), 27.2, 26.2 (2C), 25.9 (2C), 24.9 (2C), 18.4, 13.7; HRMS-ESI: m/z calcd. for C₂₇H₃₈N₄O₃S [M+H]⁺ 499.272 9, found 499.273 0.
2l	¹H NMR (400 MHz, DMSO-d₆) δ: 8.03 (d, J = 8.4 Hz, 2H, ArH), 7.84 (d, J = 8.6 Hz, 2H, ArH), 4.60 (s, 2H, CH₂-S), 3.21 (d, J = 5.5 Hz, 1H, CH-NH₂), 2.28 (q, J = 7.4 Hz, 2H, CH₂CH₃), 2.07 (d, J = 6.8 Hz, 2H, CH₂-cyclohexyl), 1.94 (dq, J = 13.4, 6.8 Hz, 1H, CH(CH₃)₂), 1.49-1.39 (m, 6H, CH(CH₃)₂), 1.30-1.20 (m, 3H, CH₂CH₃), 0.89 (m, 11H, cyclohexyl); ¹³C NMR (100 MHz, DMSO-d₆) δ: 191.8, 174.3, 163.0, 160.8, 156.1, 143.3, 130.8, 129.4 (2C), 120.8 (2C), 118.2, 60.6, 36.6, 35.9, 32.3 (2C), 31.7, 30.1, 25.7, 25.4 (2C), 19.3, 17.8, 17.2, 13.2; HRMS-ESI: m/z calcd. for C₂₆H₃₆N₄O₃S [M+H]⁺ 485.257 2, found 485.257 5.
2m	¹H NMR (400 MHz, DMSO-d₆) δ: 8.03 (d, J = 8.4 Hz, 2H, ArH), 7.83 (d, J = 8.5 Hz, 2H, ArH), 4.59 (s, 2H, CH₂-S), 3.25 (d, J = 5.6 Hz, 1H, CH-NH₂), 2.07 (d, J = 6.7 Hz, 2H, CH₂-cyclohexyl), 1.96 (m, 1H, CH(CH₃)₂), 1.80 (s, 3H, Me), 1.61 (d, J = 21.3 Hz, 1H, cyclohexyl), 1.45 (d, J = 10.2 Hz, 4H, cyclohexyl), 1.32-1.19 (m, 4H, cyclohexyl), 1.05 (t, J = 7.0 Hz, 2H, cyclohexyl), 0.93 (d, J = 6.7 Hz, 3H, CH(CH₃)₂), 0.86 (d, J = 6.7 Hz, 3H, CH(CH₃)₂); ¹³C NMR (100 MHz, DMSO-d₆) δ: 192.0, 174.0, 163.6, 161.0, 156.0, 143.4, 131.0, 129.5 (2C), 120.5 (2C), 118.3, 60.5, 36.6, 36.2 (2C), 32.4 (2C), 31.7, 25.8, 25.5 (2C), 18.5 (2C), 10.42; HRMS-ESI: m/z calcd. for C₂₅H₃₄N₄O₃S [M+H]⁺ 471.241 6, found 471.242 0.
2n	¹H NMR (400 MHz, DMSO-d₆) δ: 8.03 (d, J = 8.9 Hz, 2H, ArH), 7.89-7.79 (m, 2H, ArH), 4.58 (s, 2H, CH₂-S), 3.27-3.20 (m, 1H, CH-NH₂), 2.06 (q, J = 5.7 Hz, 2H, CH₂-cyclohexyl), 1.95 (m, 1H, CH(CH₃)₂), 1.83-1.76 (m, 3H, Me), 1.52-1.18 (m, 6H, CH(CH₃)₂), 1.04-0.79 (m, 11H, cyclohexyl); ¹³C NMR (100 MHz, DMSO-d₆) δ: 192.0, 174.1, 163.6 (2C), 156.1, 143.4, 130.9, 129.5 (2C), 118.3 (2C), 114.8, 60.6, 36.6, 36.2, 32.4 (2C), 31.7 (2C), 25.8, 25.5 (2C), 19.4 (2C), 10.4; HRMS-ESI: m/z calcd. for C₂₅H₃₄N₄O₃S [M+H]⁺ 471.241 6, found 471.241 8.
2o	¹H NMR (400 MHz, DMSO-d₆) δ: 8.02 (d, J = 8.4 Hz, 2H, ArH), 7.83 (d, J = 8.8 Hz, 2H, ArH), 4.58 (s, 2H, CH₂-S), 3.19 (d, J = 5.5 Hz, 1H, CH-NH₂), 2.28 (q, J = 7.3 Hz, 2H, CH₂CH₃), 2.07 (d, J = 6.8 Hz, 2H, CH₂-cyclohexyl), 1.98-1.88 (m, 1H, CH(CH₃)₂), 1.65-1.19 (m, 9H, CH(CH₃)₂ and CH₂CH₃), 0.88 (m, 11H, cyclohexyl); ¹³C NMR (100 MHz, DMSO-d₆) δ: 192.0, 174.5, 163.6 (2C), 156.1, 143.5, 130.9, 129.5 (2C), 118.3 (2C), 114.6, 60.8, 36.6, 36.1, 32.5 (2C), 31.8 (2C), 25.8, 25.5 (2C), 19.4, 17.9, 17.2, 13.3; HRMS-ESI: m/z calcd. for C₂₆H₃₆N₄O₃S [M+H]⁺ 485.257 2, found 485.257 0.
2p	¹H NMR (400 MHz, DMSO-d₆) δ: 8.01 (d, J = 8.7 Hz, 2H, ArH), 7.83 (d, J = 8.2 Hz, 2H, ArH), 4.57 (s, 2H, CH₂-S), 3.43-3.40 (m, 1H, CH-NH₂), 2.08 (d, J = 6.7 Hz, 2H, CH₂-cyclohexyl), 1.77-1.70 (m, 1H, CH₂CH(CH₃)₂), 1.61 (m, 8H, Me and cyclohexyl), 1.54-1.51 (m, 5H, cyclohexyl and CH₂CH(CH₃)₂), 1.17 (m, 3H, cyclohexyl), 0.91 (m, 6H, CH₂CH(CH₃)₂); ¹³C NMR (100 MHz, DMSO-d₆) δ: 192.2. 175.8. 160.9. 144.0, 131.4, 131.2, 129.9 (2C), 118.8 (2C), 112.9, 54.5, 44.4, 36.7 (2C), 33.1 (2C), 33.0, 26.3, 26.1 (2C), 23.2, 22.6 (2C), 13.8; HRMS-ESI: m/z calcd. for C₂₆H₃₆N₄O₃S [M+H]⁺ 485.257 2, found 485.257 5.
2q	¹H NMR (400 MHz, DMSO-d₆) δ: 8.06 (d, J = 8.7 Hz, 2H, ArH), 7.85 (t, J = 8.2 Hz, 2H, ArH), 4.59 (s, 2H, CH₂-S), 3.44-3.41 (m, 1H, CH-NH₂), 2.29 (m, 2H, CH₂CH₃), 2.08 (d, J = 6.7 Hz, 2H, CH₂-cyclohexyl), 1.77-1.70 (m, 1H, CH₂CH(CH₃)₂), 1.64-1.60 (m, 8H, cyclohexyl), 1.54-1.50 (m, 5H, cyclohexyl and CH₂CH(CH₃)₂), 1.17 (t, J = 7.1 Hz, 3H, CH₂CH₃), 0.92 (m, 6H, CH₂CH(CH₃)₂); ¹³C NMR (100 MHz, DMSO-d₆) δ: 192.2, 175.8, 160.9, 144.0, 131.4, 131.2 (2C), 129.9 (2C), 118.8, 112.9, 54.5, 44.4, 36.7 (2C), 33.1 (2C), 33.0, 26.3, 26.1 (2C), 23.2, 22.6 (2C), 18.5, 13.8; HRMS-ESI: m/z calc.d for C₂₇H₃₈N₄O₃S [M+H]⁺ 499.272 9, found 499.273 3.
2r	¹H NMR (400 MHz, DMSO-d₆) δ: 8.02 (d, J = 8.4 Hz, 2H, ArH), 7.83 (d, J = 8.7 Hz, 2H, ArH), 4.59 (s, 2H, CH₂-S), 3.44 (dd, J = 8.5, 5.8 Hz, 1H, CH-NH₂), 2.08 (d, J = 6.6 Hz, 2H, CH₂-cyclohexyl), 1.80 (s, 3H, Me), 1.77-1.69 (m, 1H, CH₂CH(CH₃)₂), 1.54-1.20 (m, 10H, cyclohexyl and CH₂CH(CH₃)₂), 0.89 (m, 9H, cyclohexyl and CH₂CH(CH₃)₂); ¹³C NMR (100 MHz, DMSO-d₆) δ: 192.5, 175.5, 163.3, 161.4, 156.1, 144.0, 130.9, 129.9 (2C), 120.5 (2C), 118.4, 54.2, 44.2, 37.1, 36.7, 32.8 (2C), 30.1, 25.9 (2C), 25.8, 23.5, 22.4 (2C), 13.8; HRMS-ESI: m/z calcd. for C₂₆H₃₆N₄O₃S [M+H]⁺ 485.257 2, found 485.257 6.
2s	¹H NMR (400 MHz, DMSO-d₆) δ: 8.06-7.99 (m, 2H, ArH), 7.82 (t, J = 6.2 Hz, 2H, ArH), 4.60 (d, J = 4.0 Hz, 2H, CH₂-S), 3.43-3.39 (m, 1H, CH-NH₂), 2.28 (m, 2H, CH₂CH₃), 2.08 (d, J = 5.5 Hz, 2H, CH₂-cyclohexyl), 1.73 (dp, J = 13.4, 6.6 Hz, 1H, CH₂CH(CH₃)₂), 1.51-1.23 (m, 9H, cyclohexyl and CH₂CH(CH₃)₂), 0.97-0.83 (m, 13H, cyclohexyl and CH₂CH(CH₃)₂) and CH₂CH₃); ¹³C NMR (100 MHz, DMSO-d₆) δ: 191.9, 175.3, 163.2, 160.5, 156.3, 143.6, 130.9, 129.6 (2C), 120.9 (2C), 118.4, 54.1, 43.9, 36.7, 36.1, 32.5 (2C), 25.8, 25.6 (2C), 24.2, 23.1 (2C), 21.9, 17.9, 13.3; HRMS-ESI: m/z calcd. for C₂₇H₃₈N₄O₃S [M+H]⁺ 499.272 9, found 499.272 7.
2t	¹H NMR (400 MHz, DMSO-d₆) δ: 8.02 (d, J = 8.4 Hz, 2H, ArH), 7.80 (d, J = 8.6 Hz, 2H, ArH), 7.27-7.16 (m, 5H, ArH), 4.61 (s, 2H, CH₂-S), 3.66 (m, 1H, CH-NH₂), 3.01 (dd, J = 13.4, 5.7 Hz, 1H, CH₂-Ph), 2.77 (dd, J = 13.4, 7.7 Hz, 1H, CH₂-Ph), 2.09 (d, J = 6.6 Hz, 2H, CH₂-cyclohexyl), 1.80 (s, 3H, Me), 1.26 (t, J = 14.8 Hz, 7H, cyclohexyl), 0.92 (h, J = 13.4, 12.3 Hz, 4H, cyclohexyl); ¹³C NMR (100 MHz, DMSO-d₆) δ: 192.3, 174.4, 163.9 (2C), 156.4, 143.9, 138.7, 131.4, 130.0, 129.7 (2C), 128.5 (2C), 126.6 (2C), 118.8 (2C), 115.4, 57.5, 37.1, 36.7 (2C), 32.9 (2C), 26.3, 26.0 (3C), 10.8; HRMS-ESI: m/z calcd. for C₂₉H₃₄N₄O₃S [M+H]⁺ 519.241 6, found 519.242 0.
2u	¹H NMR (400 MHz, DMSO-d₆) δ: 8.02 (d, J = 8.4 Hz, 2H, ArH), 7.81 (d, J = 8.4 Hz, 2H, ArH), 7.26-7.16 (m, 5H, ArH), 4.61 (s, 2H, CH₂-S), 3.69 (m, 1H, CH-NH₂), 3.02 (dd, J = 13.4, 5.7 Hz, 1H, CH₂-Ph), 2.77 (dd, J = 13.4, 7.7 Hz, 1H, CH₂-Ph), 2.29 (q, J = 7.3 Hz, 2H, CH₂CH₃), 2.08 (d, J = 6.8 Hz, 2H, CH₂-cyclohexyl), 1.52-1.18 (m, 7H, cyclohexyl), 0.91 (t, J = 7.6 Hz, 7H, cyclohexyl and CH₂CH₃); ¹³C NMR (100 MHz, DMSO-d₆) δ: 192.3, 174.4, 163.6, 160.9, 156.7, 143.9, 138.7, 131.4, 129.9 (2C), 129.8 (2C), 128.5 (2C), 126.6, 121.4 (2C), 118.9, 57.5, 37.2, 36.6 (2C), 32.9 (2C), 26.3, 26.0 (3C), 18.4, 13.7; HRMS-ESI: m/z calcd. for C₃₀H₃₆N₄O₃S [M+H]⁺ 533.257 2, found 533.257 0.
2v	¹H NMR (400 MHz, DMSO-d₆) δ: 10.44 (s, 1H, C=O-NH), 8.02 (d, J = 8.4 Hz, 2H, ArH), 7.85 (d, J = 8.5 Hz, 2H, ArH), 4.59 (s, 2H, CH₂-S), 3.80 (m, 1H, pyrrolidinyl), 2.92 (t, J = 6.6 Hz, 2H, pyrrolidinyl), 2.08 (d, J = 7.0 Hz, 2H, CH₂-cyclohexyl), 1.79 (s, 3H, Me), 1.73-1.18 (m, 11H, cyclohexyl), 0.91 (m, 4H, pyrrolidinyl); ¹³C NMR (100 MHz, DMSO-d₆) δ: 192.5, 174.0, 164.1, 161.4, 156.5, 143.5, 131.6, 130.0 (2C), 118.8 (2C), 115.4, 61.2, 47.1, 37.1, 36.7, 32.8 (2C), 30.9 (2C), 26.3, 26.1 (2C), 25.9, 10.9; HRMS-ESI: m/z calcd. for C₂₅H₃₂N₄O₃S [M+H]⁺ 469.225 9, found 469.226 3.
2w	¹H NMR (400 MHz, DMSO-d₆) δ: 10.69 (s, 1H, C=O-NH), 8.20-7.70 (m, 4H, ArH), 4.59 (s, 2H, CH₂-S), 3.90 (s, 1H, pyrrolidinyl), 2.95 (m, 2H, pyrrolidinyl), 2.16 (m, 4H, CH₂-cyclohexyl and CH₂CH₃), 1.72-1.10 (m, 12H, cyclohexyl and pyrrolidinyl), 1.01-0.75 (m, 6H, pyrrolidinyl and CH₂CH₃); ¹³C NMR (100 MHz, DMSO-d₆) δ: 192.5, 173.2, 163.8, 160.8, 156.8, 143.5, 131.6, 129.9 (2C), 121.3 (2C), 118.8, 61.0, 46.9, 43.9, 37.2, 36.5, 32.9, 30.8, 26.3, 25.9, 25.8, 22.5, 22.2, 18.4, 13.7; HRMS-ESI: m/z calcd. for C₂₆H₃₄N₄O₃S [M+H]⁺ 483.241 6, found 483.241 8.
2x	¹H NMR (400 MHz, DMSO-d₆) δ: 10.69 (s, 1H, C=O-NH), 8.20 (d, J = 8.4 Hz, 2H, ArH), 7.80 (d, J = 8.6 Hz, 2H, ArH), 4.60 (s, 2H, CH₂-S), 3.90 (t, 1H, pyrrolidinyl), 2.95 (m, 2H, pyrrolidinyl), 2.16 (q, J = 8.5 Hz, 2H, CH₂CH₃), 2.06(d, J = 7.2, 2H, CH₂-cyclohexyl), 1.72-1.10 (m, 12H, cyclohexyl and pyrrolidinyl), 1.01-0.75 (m, 6H, pyrrolidinyl and CH₂CH₃); ¹³C NMR (100 MHz, DMSO-d₆) δ: 191.4, 172.2, 163.8, 159.8, 156.8, 146.2, 131.8, 129.9 (2C), 121.3 (2C), 118.8, 62.4, 46.9, 43.9, 37.2, 36.5, 32.9, 30.8, 26.3, 26.0, 25.8, 22.6, 22.3, 18.4, 14.0; HRMS-ESI: m/z calcd. for C₂₆H₃₄N₄O₃S [M+H]⁺ 483.241 6, found 483.242 0.

Compd.	¹H NMR, ¹³C NMR, HRMS-ESI
2a	¹H NMR (400 MHz, DMSO-d₆) δ: 8.02 (d, J = 8.5 Hz, 2H, ArH), 7.86 (d, J = 9.2 Hz, 2H, ArH), 4.60 (s, 2H, CH₂-S), 3.61 (d, J = 2.9 Hz, 2H, NH₂CH₂), 2.09 (d, J = 6.7 Hz, 2H, CH₂-cyclohexyl), 1.79 (s, 3H, Me), 1.66 (m, 7H, Cyclohexyl), 1.54 (m, 4H, cyclohexyl); ¹³C NMR (100 MHz, DMSO-d₆) δ: 190.3, 174.3, 161.5 (2C), 155.9, 142.8, 131.1, 130.0 (2C), 118.7 (2C), 115.3, 43.9, 37.6 (2C), 36.8 (2C), 32.9, 32.6 (3C), 10.9; HRMS-ESI: m/z calcd. for C₂₂H₂₈N₄O₃S [M+H]⁺ 429.194 6, found 429.194 8.
2b	¹H NMR (400 MHz, DMSO-d₆) δ: 8.03 (d, J = 8.5 Hz, 2H, ArH), 7.83 (d, J = 9.9 Hz, 2H, ArH), 4.60 (s, 2H, CH₂-S), 3.60 (d, J = 2.9 Hz, 2H, NH₂CH₂), 2.29-2.26 (m, 2H, CH₂CH₃), 2.08 (d, J = 6.9 Hz, 2H, CH₂-cyclohexyl), 1.65 (m, 7H, cyclohexyl), 1.53 (dd, J = 6.1, 2.1 Hz, 4H, cyclohexyl), 0.91 (s, 3H, CH₂CH₃); ¹³C NMR (100 MHz, DMSO-d₆) δ: 192.3, 174.4, 161.7 (2C), 156.4, 143.2, 131.0, 130.0 (2C), 118.7 (2C), 117.2, 43.9, 37.6, 36.6 (2C), 32.9 (2C), 32.6 (3C), 22.5, 13.7; HRMS-ESI: m/z calcd. for C₂₃H₃₀N₄O₃S [M+H]⁺ 443.210 3, found 443.210 5.
2c	¹H NMR (400 MHz, DMSO-d₆) δ: 8.02 (s, 2H, ArH), 7.88-7.79 (m, 2H, ArH), 4.63-4.55 (m, 2H, CH₂-S), 3.62-3.55 (m, 1H, CH-NH₂), 2.15-2.04 (m, 2H, CH₂-cyclohexyl), 1.83-1.77 (m, 3H, Me), 1.68-1.60 (m, 1H, cyclohexyl), 1.54-1.48 (m, 3H, CHCH₃), 1.35-1.21 (m, 7H, cyclohexyl), 1.02-0.90 (m, 3H, cyclohexyl); ¹³C NMR (100 MHz, DMSO-d₆) δ: 191.9, 174.7, 164.1, 160.9, 156.4, 143.4, 131.0, 129.5 (2C), 118.3 (2C), 114.7, 50.87, 36.6, 36.3, 32.4 (2C), 25.8, 25.5 (3C), 20.7, 10.4; HRMS-ESI: m/z calcd. for C₂₃H₃₀N₄O₃S [M+H]⁺ 443.210 3, found 443.210 6.
2d	¹H NMR (400 MHz, DMSO-d₆) δ: 8.02 (d, J = 8.4 Hz, 2H, ArH), 7.82 (d, J = 8.5 Hz, 2H, ArH), 4.60 (s, 2H, CH₂-S), 3.54 (q, J = 7.1 Hz, 1H, CH-NH₂), 2.28 (q, J = 7.4 Hz, 2H, CH₂CH₃), 2.08 (d, J = 6.7 Hz, 2H, CH₂-cyclohexyl), 1.50-1.40 (m, 3H, CHCH₃), 1.36-1.28 (m, 2H, cyclohexyl), 1.25 (t, J = 6.1 Hz, 5H, cyclohexyl), 0.91 (t, J = 7.2 Hz, 7H, cyclohexyl and CH₂CH₃); ¹³C NMR (100 MHz, DMSO-d₆) δ: 191.9, 174.9, 163.2, 160.44, 156.32, 143.51, 130.96, 129.55 (2C), 118.33 (2C), 114.63, 50.95, 36.76, 36.14, 32.50 (2C), 25.83, 25.55 (3C), 20.87, 17.97, 13.27; HRMS-ESI: m/z calcd. for C₂₄H₃₂N₄O₃S [M+H]⁺ 457.225 9, found 457.226 1.
2e	¹H NMR (400 MHz, DMSO-d₆) δ: 8.07 (s, 2H, ArH), 7.86-7.72 (m, 2H, ArH), 4.60 (s, 2H, CH₂-S), 3.72 (m, 1H, CH-NH₂), 2.15-2.01 (m, 2H, CH₂-cyclohexyl), 1.80-1.74 (s, 3H, Me), 1.71-1.62 (m, 1H, cyclohexyl), 1.52-1.44 (m, 3H, CHCH₃), 1.36-1.09 (m, 7H, cyclohexyl), 1.03-0.90 (m, 3H, cyclohexyl); ¹³C NMR (100 MHz, DMSO-d₆) δ: 191.6, 174.7, 163.2, 160.5, 154.1, 144.0, 131.8, 129.0 (2C), 117.8 (2C), 114.0, 51.0, 36.8, 36.3, 32.8 (2C), 25.9, 25.3 (3C), 18.8, 10.3; HRMS-ESI: m/z calcd. for C₂₃H₃₀N₄O₃S [M+H]⁺ 443.210 3, found 443.210 7.
2f	¹H NMR (400 MHz, DMSO-d₆) δ: 8.04 (d, J = 8.4 Hz, 2H, ArH), 7.80 (d, J = 8.5 Hz, 2H, ArH), 4.61 (s, 2H, CH₂-S), 3.60 (q, J = 7.8 Hz, 1H, CH-NH₂), 2.18 (q, J = 7.2 Hz, 2H, CH₂CH₃), 2.08 (d, J = 7.0 Hz, 2H, CH₂-cyclohexyl), 1.56-1.40 (m, 3H, CHCH₃), 1.35-1.26 (m, 2H, cyclohexyl), 1.21 (t, J = 6.1 Hz, 5H, cyclohexyl), 0.91 (t, J = 7.2 Hz, 7H, cyclohexyl and CH₂CH₃); ¹³C NMR (100 MHz, DMSO-d₆) δ: 191.2, 174.6, 163.8, 161.1, 156.3, 144.7, 131.0, 129.5 (2C), 118.1 (2C), 114.3, 51.8, 36.9 (2C), 35.4 (2C), 31.6, 26.6 (2C), 25.1, 20.2, 18.3, 14.1; HRMS-ESI: m/z calcd. for C₂₄H₃₂N₄O₃S [M+H]⁺ 457.225 9, found 457.226 3.
2g	¹H NMR (400 MHz, DMSO-d₆) δ: 8.09 (d, J = 8.4 Hz, 2H, ArH), 7.82 (d, J = 8.5 Hz, 2H, ArH), 4.68 (s, 2H, CH₂-S), 3.11 (m, 2H, NH₂CH₂CH₂), 2.60 (m, 2H, NH₂CH₂CH₂), 2.28 (m, 2H, CH₂CH₃), 2.08 (d, J = 7.8 Hz, 2H, CH₂-cyclohexyl), 1.56-1.29 (m, 11H, cyclohexyl), 0.91 (t, J = 7.2 Hz, 3H, CH₂CH₃); ¹³C NMR (100 MHz, DMSO-d₆) δ: 191.6, 173.6, 162.1, 160.8, 157.9, 143.6, 131.0, 129.6 (2C), 119.2 (2C), 114.3, 39.8, 36.9, 36.0, 34.5, 33.8 (2C), 32.7, 26.4, 25.3 (2C), 20.2, 14.2; HRMS-ESI: m/z calcd. for C₂₄H₃₂N₄O₃S [M+H]⁺ 457.225 9, found 457.226 0.
2h	¹H NMR (400 MHz, DMSO-d₆) δ: 8.02 (d, J = 8.5 Hz, 2H, ArH), 7.82 (d, J = 8.8 Hz, 2H, ArH), 4.58 (s, 2H, CH₂-S), 3.40 (t, J = 6.6 Hz, 1H, CH-NH₂), 2.07 (s, 3H, Me), 1.79 (m, 4H, CH₂-cyclohexyl and CH₂CH₂CH₂CH₃), 1.66-0.91 (m, 15H, cyclohexyl and CH₂CH₂CH₂CH₃), 0.84 (t, J = 6.9 Hz, 3H, CH₂CH₂CH₂CH₃); ¹³C NMR (100 MHz, DMSO-d₆) δ: 192.0, 174.7, 163.7, 161.0, 156.2, 143.5, 131.0, 129.5 (2C), 118.3 (2C), 114.9, 55.4, 36.6, 36.2, 34.4, 32.4 (2C), 27.3, 25.8 (3C), 25.5, 22.1, 13.9, 10.4; HRMS-ESI: m/z calcd. for C₂₆H₃₆N₄O₃S [M+H]⁺ 485.257 2, found 485.256 8.
2i	¹H NMR (400 MHz, DMSO-d₆) δ: 8.06 (d, J = 8.4 Hz, 2H, ArH), 7.84 (d, J = 8.7 Hz, 2H, ArH), 4.59 (s, 2H, CH₂-S), 3.37 (t, J = 6.5 Hz, 1H, CH-NH₂), 2.28 (q, J = 7.3 Hz, 2H, CH₂CH₃), 2.06 (d, J = 6.7 Hz, 2H, CH₂-cyclohexyl), 1.69-1.17 (m, 13H, cyclohexyl and CH₂CH₂CH₂CH₃), 0.88 (dt, J = 25.0, 7.2 Hz, 10H, CH₂CH₃ and CH₂CH₂CH₂CH₃); ¹³C NMR (100 MHz, DMSO-d₆) δ: 191.9, 174.8, 163.1 (2C), 156.2, 143.5, 130.9, 129.5 (2C), 120.9 (2C), 118.3, 55.5, 36.7, 36.1, 34.5, 32.4 (2C), 27.4, 25.8, 25.5 (2C), 22.1, 17.9, 13.8, 13.2; HRMS-ESI: m/z calcd. for C₂₇H₃₈N₄O₃S [M+H]⁺ 499.272 9, found 499.272 6.
2j	¹H NMR (400 MHz, DMSO-d₆) δ: 8.02 (d, J = 8.4 Hz, 2H, ArH), 7.82 (d, J = 8.8 Hz, 2H, ArH), 4.63 (s, 1H, CH₂-S), 2.78 (t, J = 7.6 Hz, 2H, NH₂CH₂CH₂CH₂CH₂CH₂), 2.40 (t, J = 7.3 Hz, 2H, NH₂CH₂CH₂CH₂CH₂CH₂), 2.28 (d, J = 7.6 Hz, 2H, CH₂-cyclohexyl), 2.08 (s, 3H, Me), 1.66 (q, J = 5.8 Hz, 4H, NH₂CH₂CH₂CH₂CH₂CH₂), 1.61-1.57 (m, 4H, cyclohexyl and NH₂CH₂CH₂CH₂CH₂CH₂), 1.57-1.50 (m, 4H, cyclohexyl), 1.37-1.25 (m, 5H, cyclohexyl); ¹³C NMR (100 MHz, DMSO-d₆) δ: 192.1, 175.5, 164.1 (2C), 155.3, 144.2, 131.0, 129.8 (2C), 118.0 (2C), 117.0, 43, 0, 37.1, 32.9 (2C), 27.2, 26.4 (2C), 25.7 (3C), 24.6, 13.7; HRMS-ESI: m/z calcd. for C₂₆H₃₆N₄O₃S [M+H]⁺ 485.257 2, found 485.257 7.
2k	¹H NMR (400 MHz, DMSO-d₆) δ: 8.00 (d, J = 8.5 Hz, 2H, ArH), 7.82 (d, J = 8.6 Hz, 2H, ArH), 4.60 (s, 2H, CH₂-S), 2.76 (t, J = 7.6 Hz, 2H, NH₂CH₂CH₂CH₂CH₂CH₂), 2.40 (t, J = 7.3 Hz, 2H, NH₂CH₂CH₂CH₂CH₂CH₂), 2.30 (m, 2H, CH₂CH₃), 2.08 (d, J = 6.8 Hz, 2H, CH₂-cyclohexyl), 1.66 (q, J = 5.8 Hz, 4H, NH₂CH₂CH₂CH₂CH₂CH₂), 1.61-1.57 (m, 4H, cyclohexyl), 1.57-1.50 (m, 4H, cyclohexyl and NH₂CH₂CH₂CH₂CH₂CH₂), 1.37-1.25 (m, 5H, cyclohexyl), 0.92 (m, 3H, CH₂CH₃); ¹³C NMR (100 MHz, DMSO-d₆) δ: 192.3, 175.8, 164.3 (2C), 155.2, 144.5, 131.1, 129.9 (2C), 118.6 (2C), 117.2, 42.8, 37.2 (2C), 32.9 (2C), 27.2, 26.2 (2C), 25.9 (2C), 24.9 (2C), 18.4, 13.7; HRMS-ESI: m/z calcd. for C₂₇H₃₈N₄O₃S [M+H]⁺ 499.272 9, found 499.273 0.
2l	¹H NMR (400 MHz, DMSO-d₆) δ: 8.03 (d, J = 8.4 Hz, 2H, ArH), 7.84 (d, J = 8.6 Hz, 2H, ArH), 4.60 (s, 2H, CH₂-S), 3.21 (d, J = 5.5 Hz, 1H, CH-NH₂), 2.28 (q, J = 7.4 Hz, 2H, CH₂CH₃), 2.07 (d, J = 6.8 Hz, 2H, CH₂-cyclohexyl), 1.94 (dq, J = 13.4, 6.8 Hz, 1H, CH(CH₃)₂), 1.49-1.39 (m, 6H, CH(CH₃)₂), 1.30-1.20 (m, 3H, CH₂CH₃), 0.89 (m, 11H, cyclohexyl); ¹³C NMR (100 MHz, DMSO-d₆) δ: 191.8, 174.3, 163.0, 160.8, 156.1, 143.3, 130.8, 129.4 (2C), 120.8 (2C), 118.2, 60.6, 36.6, 35.9, 32.3 (2C), 31.7, 30.1, 25.7, 25.4 (2C), 19.3, 17.8, 17.2, 13.2; HRMS-ESI: m/z calcd. for C₂₆H₃₆N₄O₃S [M+H]⁺ 485.257 2, found 485.257 5.
2m	¹H NMR (400 MHz, DMSO-d₆) δ: 8.03 (d, J = 8.4 Hz, 2H, ArH), 7.83 (d, J = 8.5 Hz, 2H, ArH), 4.59 (s, 2H, CH₂-S), 3.25 (d, J = 5.6 Hz, 1H, CH-NH₂), 2.07 (d, J = 6.7 Hz, 2H, CH₂-cyclohexyl), 1.96 (m, 1H, CH(CH₃)₂), 1.80 (s, 3H, Me), 1.61 (d, J = 21.3 Hz, 1H, cyclohexyl), 1.45 (d, J = 10.2 Hz, 4H, cyclohexyl), 1.32-1.19 (m, 4H, cyclohexyl), 1.05 (t, J = 7.0 Hz, 2H, cyclohexyl), 0.93 (d, J = 6.7 Hz, 3H, CH(CH₃)₂), 0.86 (d, J = 6.7 Hz, 3H, CH(CH₃)₂); ¹³C NMR (100 MHz, DMSO-d₆) δ: 192.0, 174.0, 163.6, 161.0, 156.0, 143.4, 131.0, 129.5 (2C), 120.5 (2C), 118.3, 60.5, 36.6, 36.2 (2C), 32.4 (2C), 31.7, 25.8, 25.5 (2C), 18.5 (2C), 10.42; HRMS-ESI: m/z calcd. for C₂₅H₃₄N₄O₃S [M+H]⁺ 471.241 6, found 471.242 0.
2n	¹H NMR (400 MHz, DMSO-d₆) δ: 8.03 (d, J = 8.9 Hz, 2H, ArH), 7.89-7.79 (m, 2H, ArH), 4.58 (s, 2H, CH₂-S), 3.27-3.20 (m, 1H, CH-NH₂), 2.06 (q, J = 5.7 Hz, 2H, CH₂-cyclohexyl), 1.95 (m, 1H, CH(CH₃)₂), 1.83-1.76 (m, 3H, Me), 1.52-1.18 (m, 6H, CH(CH₃)₂), 1.04-0.79 (m, 11H, cyclohexyl); ¹³C NMR (100 MHz, DMSO-d₆) δ: 192.0, 174.1, 163.6 (2C), 156.1, 143.4, 130.9, 129.5 (2C), 118.3 (2C), 114.8, 60.6, 36.6, 36.2, 32.4 (2C), 31.7 (2C), 25.8, 25.5 (2C), 19.4 (2C), 10.4; HRMS-ESI: m/z calcd. for C₂₅H₃₄N₄O₃S [M+H]⁺ 471.241 6, found 471.241 8.
2o	¹H NMR (400 MHz, DMSO-d₆) δ: 8.02 (d, J = 8.4 Hz, 2H, ArH), 7.83 (d, J = 8.8 Hz, 2H, ArH), 4.58 (s, 2H, CH₂-S), 3.19 (d, J = 5.5 Hz, 1H, CH-NH₂), 2.28 (q, J = 7.3 Hz, 2H, CH₂CH₃), 2.07 (d, J = 6.8 Hz, 2H, CH₂-cyclohexyl), 1.98-1.88 (m, 1H, CH(CH₃)₂), 1.65-1.19 (m, 9H, CH(CH₃)₂ and CH₂CH₃), 0.88 (m, 11H, cyclohexyl); ¹³C NMR (100 MHz, DMSO-d₆) δ: 192.0, 174.5, 163.6 (2C), 156.1, 143.5, 130.9, 129.5 (2C), 118.3 (2C), 114.6, 60.8, 36.6, 36.1, 32.5 (2C), 31.8 (2C), 25.8, 25.5 (2C), 19.4, 17.9, 17.2, 13.3; HRMS-ESI: m/z calcd. for C₂₆H₃₆N₄O₃S [M+H]⁺ 485.257 2, found 485.257 0.
2p	¹H NMR (400 MHz, DMSO-d₆) δ: 8.01 (d, J = 8.7 Hz, 2H, ArH), 7.83 (d, J = 8.2 Hz, 2H, ArH), 4.57 (s, 2H, CH₂-S), 3.43-3.40 (m, 1H, CH-NH₂), 2.08 (d, J = 6.7 Hz, 2H, CH₂-cyclohexyl), 1.77-1.70 (m, 1H, CH₂CH(CH₃)₂), 1.61 (m, 8H, Me and cyclohexyl), 1.54-1.51 (m, 5H, cyclohexyl and CH₂CH(CH₃)₂), 1.17 (m, 3H, cyclohexyl), 0.91 (m, 6H, CH₂CH(CH₃)₂); ¹³C NMR (100 MHz, DMSO-d₆) δ: 192.2. 175.8. 160.9. 144.0, 131.4, 131.2, 129.9 (2C), 118.8 (2C), 112.9, 54.5, 44.4, 36.7 (2C), 33.1 (2C), 33.0, 26.3, 26.1 (2C), 23.2, 22.6 (2C), 13.8; HRMS-ESI: m/z calcd. for C₂₆H₃₆N₄O₃S [M+H]⁺ 485.257 2, found 485.257 5.
2q	¹H NMR (400 MHz, DMSO-d₆) δ: 8.06 (d, J = 8.7 Hz, 2H, ArH), 7.85 (t, J = 8.2 Hz, 2H, ArH), 4.59 (s, 2H, CH₂-S), 3.44-3.41 (m, 1H, CH-NH₂), 2.29 (m, 2H, CH₂CH₃), 2.08 (d, J = 6.7 Hz, 2H, CH₂-cyclohexyl), 1.77-1.70 (m, 1H, CH₂CH(CH₃)₂), 1.64-1.60 (m, 8H, cyclohexyl), 1.54-1.50 (m, 5H, cyclohexyl and CH₂CH(CH₃)₂), 1.17 (t, J = 7.1 Hz, 3H, CH₂CH₃), 0.92 (m, 6H, CH₂CH(CH₃)₂); ¹³C NMR (100 MHz, DMSO-d₆) δ: 192.2, 175.8, 160.9, 144.0, 131.4, 131.2 (2C), 129.9 (2C), 118.8, 112.9, 54.5, 44.4, 36.7 (2C), 33.1 (2C), 33.0, 26.3, 26.1 (2C), 23.2, 22.6 (2C), 18.5, 13.8; HRMS-ESI: m/z calc.d for C₂₇H₃₈N₄O₃S [M+H]⁺ 499.272 9, found 499.273 3.
2r	¹H NMR (400 MHz, DMSO-d₆) δ: 8.02 (d, J = 8.4 Hz, 2H, ArH), 7.83 (d, J = 8.7 Hz, 2H, ArH), 4.59 (s, 2H, CH₂-S), 3.44 (dd, J = 8.5, 5.8 Hz, 1H, CH-NH₂), 2.08 (d, J = 6.6 Hz, 2H, CH₂-cyclohexyl), 1.80 (s, 3H, Me), 1.77-1.69 (m, 1H, CH₂CH(CH₃)₂), 1.54-1.20 (m, 10H, cyclohexyl and CH₂CH(CH₃)₂), 0.89 (m, 9H, cyclohexyl and CH₂CH(CH₃)₂); ¹³C NMR (100 MHz, DMSO-d₆) δ: 192.5, 175.5, 163.3, 161.4, 156.1, 144.0, 130.9, 129.9 (2C), 120.5 (2C), 118.4, 54.2, 44.2, 37.1, 36.7, 32.8 (2C), 30.1, 25.9 (2C), 25.8, 23.5, 22.4 (2C), 13.8; HRMS-ESI: m/z calcd. for C₂₆H₃₆N₄O₃S [M+H]⁺ 485.257 2, found 485.257 6.
2s	¹H NMR (400 MHz, DMSO-d₆) δ: 8.06-7.99 (m, 2H, ArH), 7.82 (t, J = 6.2 Hz, 2H, ArH), 4.60 (d, J = 4.0 Hz, 2H, CH₂-S), 3.43-3.39 (m, 1H, CH-NH₂), 2.28 (m, 2H, CH₂CH₃), 2.08 (d, J = 5.5 Hz, 2H, CH₂-cyclohexyl), 1.73 (dp, J = 13.4, 6.6 Hz, 1H, CH₂CH(CH₃)₂), 1.51-1.23 (m, 9H, cyclohexyl and CH₂CH(CH₃)₂), 0.97-0.83 (m, 13H, cyclohexyl and CH₂CH(CH₃)₂) and CH₂CH₃); ¹³C NMR (100 MHz, DMSO-d₆) δ: 191.9, 175.3, 163.2, 160.5, 156.3, 143.6, 130.9, 129.6 (2C), 120.9 (2C), 118.4, 54.1, 43.9, 36.7, 36.1, 32.5 (2C), 25.8, 25.6 (2C), 24.2, 23.1 (2C), 21.9, 17.9, 13.3; HRMS-ESI: m/z calcd. for C₂₇H₃₈N₄O₃S [M+H]⁺ 499.272 9, found 499.272 7.
2t	¹H NMR (400 MHz, DMSO-d₆) δ: 8.02 (d, J = 8.4 Hz, 2H, ArH), 7.80 (d, J = 8.6 Hz, 2H, ArH), 7.27-7.16 (m, 5H, ArH), 4.61 (s, 2H, CH₂-S), 3.66 (m, 1H, CH-NH₂), 3.01 (dd, J = 13.4, 5.7 Hz, 1H, CH₂-Ph), 2.77 (dd, J = 13.4, 7.7 Hz, 1H, CH₂-Ph), 2.09 (d, J = 6.6 Hz, 2H, CH₂-cyclohexyl), 1.80 (s, 3H, Me), 1.26 (t, J = 14.8 Hz, 7H, cyclohexyl), 0.92 (h, J = 13.4, 12.3 Hz, 4H, cyclohexyl); ¹³C NMR (100 MHz, DMSO-d₆) δ: 192.3, 174.4, 163.9 (2C), 156.4, 143.9, 138.7, 131.4, 130.0, 129.7 (2C), 128.5 (2C), 126.6 (2C), 118.8 (2C), 115.4, 57.5, 37.1, 36.7 (2C), 32.9 (2C), 26.3, 26.0 (3C), 10.8; HRMS-ESI: m/z calcd. for C₂₉H₃₄N₄O₃S [M+H]⁺ 519.241 6, found 519.242 0.
2u	¹H NMR (400 MHz, DMSO-d₆) δ: 8.02 (d, J = 8.4 Hz, 2H, ArH), 7.81 (d, J = 8.4 Hz, 2H, ArH), 7.26-7.16 (m, 5H, ArH), 4.61 (s, 2H, CH₂-S), 3.69 (m, 1H, CH-NH₂), 3.02 (dd, J = 13.4, 5.7 Hz, 1H, CH₂-Ph), 2.77 (dd, J = 13.4, 7.7 Hz, 1H, CH₂-Ph), 2.29 (q, J = 7.3 Hz, 2H, CH₂CH₃), 2.08 (d, J = 6.8 Hz, 2H, CH₂-cyclohexyl), 1.52-1.18 (m, 7H, cyclohexyl), 0.91 (t, J = 7.6 Hz, 7H, cyclohexyl and CH₂CH₃); ¹³C NMR (100 MHz, DMSO-d₆) δ: 192.3, 174.4, 163.6, 160.9, 156.7, 143.9, 138.7, 131.4, 129.9 (2C), 129.8 (2C), 128.5 (2C), 126.6, 121.4 (2C), 118.9, 57.5, 37.2, 36.6 (2C), 32.9 (2C), 26.3, 26.0 (3C), 18.4, 13.7; HRMS-ESI: m/z calcd. for C₃₀H₃₆N₄O₃S [M+H]⁺ 533.257 2, found 533.257 0.
2v	¹H NMR (400 MHz, DMSO-d₆) δ: 10.44 (s, 1H, C=O-NH), 8.02 (d, J = 8.4 Hz, 2H, ArH), 7.85 (d, J = 8.5 Hz, 2H, ArH), 4.59 (s, 2H, CH₂-S), 3.80 (m, 1H, pyrrolidinyl), 2.92 (t, J = 6.6 Hz, 2H, pyrrolidinyl), 2.08 (d, J = 7.0 Hz, 2H, CH₂-cyclohexyl), 1.79 (s, 3H, Me), 1.73-1.18 (m, 11H, cyclohexyl), 0.91 (m, 4H, pyrrolidinyl); ¹³C NMR (100 MHz, DMSO-d₆) δ: 192.5, 174.0, 164.1, 161.4, 156.5, 143.5, 131.6, 130.0 (2C), 118.8 (2C), 115.4, 61.2, 47.1, 37.1, 36.7, 32.8 (2C), 30.9 (2C), 26.3, 26.1 (2C), 25.9, 10.9; HRMS-ESI: m/z calcd. for C₂₅H₃₂N₄O₃S [M+H]⁺ 469.225 9, found 469.226 3.
2w	¹H NMR (400 MHz, DMSO-d₆) δ: 10.69 (s, 1H, C=O-NH), 8.20-7.70 (m, 4H, ArH), 4.59 (s, 2H, CH₂-S), 3.90 (s, 1H, pyrrolidinyl), 2.95 (m, 2H, pyrrolidinyl), 2.16 (m, 4H, CH₂-cyclohexyl and CH₂CH₃), 1.72-1.10 (m, 12H, cyclohexyl and pyrrolidinyl), 1.01-0.75 (m, 6H, pyrrolidinyl and CH₂CH₃); ¹³C NMR (100 MHz, DMSO-d₆) δ: 192.5, 173.2, 163.8, 160.8, 156.8, 143.5, 131.6, 129.9 (2C), 121.3 (2C), 118.8, 61.0, 46.9, 43.9, 37.2, 36.5, 32.9, 30.8, 26.3, 25.9, 25.8, 22.5, 22.2, 18.4, 13.7; HRMS-ESI: m/z calcd. for C₂₆H₃₄N₄O₃S [M+H]⁺ 483.241 6, found 483.241 8.
2x	¹H NMR (400 MHz, DMSO-d₆) δ: 10.69 (s, 1H, C=O-NH), 8.20 (d, J = 8.4 Hz, 2H, ArH), 7.80 (d, J = 8.6 Hz, 2H, ArH), 4.60 (s, 2H, CH₂-S), 3.90 (t, 1H, pyrrolidinyl), 2.95 (m, 2H, pyrrolidinyl), 2.16 (q, J = 8.5 Hz, 2H, CH₂CH₃), 2.06(d, J = 7.2, 2H, CH₂-cyclohexyl), 1.72-1.10 (m, 12H, cyclohexyl and pyrrolidinyl), 1.01-0.75 (m, 6H, pyrrolidinyl and CH₂CH₃); ¹³C NMR (100 MHz, DMSO-d₆) δ: 191.4, 172.2, 163.8, 159.8, 156.8, 146.2, 131.8, 129.9 (2C), 121.3 (2C), 118.8, 62.4, 46.9, 43.9, 37.2, 36.5, 32.9, 30.8, 26.3, 26.0, 25.8, 22.6, 22.3, 18.4, 14.0; HRMS-ESI: m/z calcd. for C₂₆H₃₄N₄O₃S [M+H]⁺ 483.241 6, found 483.242 0.

Compd.	R¹	CC₅₀ /µmol·L^-1	EC₅₀ /µmol·L^-1	TI
2a	Me	> 200.00	2.59 ± 0.95	> 77.31
2b	Et	> 200.00	0.15 ± 0.02	> 1 375.09
2c	Me	17.32 ± 2.60	0.09 ± 0.01	197.78
2d	Et	47.52 ± 0.25	0.02 ± 0.01	2 159.67
2e	Me	150.60 ± 6.09	1.54 ± 0.05	97.76
2f	Et	> 200.00	0.47 ± 0.12	> 429.78
2g	Et	106.68 ± 17.93	0.87 ± 0.18	122.09
2h	Me	41.17 ± 2.03	0.13 ± 0.05	316.79
2i	Et	50.53 ± 8.03	0.02 ± 0.01	2 797.55
2j	Me	69.70 ± 5.40	4.04 ± 0.79	17.27
2k	Et	89.31 ± 4.39	0.13 ± 0.03	684.40
2l	Et	22.61 ± 3.11	0.03 ± 0.00	916.56
2m	Me	31.75 ± 0.46	0.55 ± 0.02	57.99
2n	Me	15.70 ± 1.41	0.08 ± 0.00	187.32
2o	Et	62.09 ± 10.20	0.06 ± 0.01	1 014.93
2p	Me	74.80 ± 9.31	0.66 ± 0.18	112.94
2q	Et	66.34 ± 4.12	0.12 ± 0.01	550.22
2r	Me	45.15 ± 4.28	0.11 ± 0.06	401.64
2s	Et	55.80 ± 1.46	0.02 ± 0.01	3 401.77
2t	Me	18.77 ± 0.42	0.23 ± 0.02	80.04
2u	Et	35.03 ± 10.07	0.05 ± 0.01	663.13
2v	Me	24.74 ± 1.18	0.27 ± 0.02	91.20
2w	Et	59.62 ± 0.79	0.01 ± 0.01	4 881.85
2x	Et	44.30 ± 8.79	0.13 ± 0.07	333.42
DB02		> 200.00	0.03 ± 0.01	> 7 750.81
3TC		> 200.00	0.61 ± 0.72	> 325.42

Compd.	R¹	CC₅₀ /µmol·L^-1	EC₅₀ /µmol·L^-1	TI
2a	Me	> 200.00	2.59 ± 0.95	> 77.31
2b	Et	> 200.00	0.15 ± 0.02	> 1 375.09
2c	Me	17.32 ± 2.60	0.09 ± 0.01	197.78
2d	Et	47.52 ± 0.25	0.02 ± 0.01	2 159.67
2e	Me	150.60 ± 6.09	1.54 ± 0.05	97.76
2f	Et	> 200.00	0.47 ± 0.12	> 429.78
2g	Et	106.68 ± 17.93	0.87 ± 0.18	122.09
2h	Me	41.17 ± 2.03	0.13 ± 0.05	316.79
2i	Et	50.53 ± 8.03	0.02 ± 0.01	2 797.55
2j	Me	69.70 ± 5.40	4.04 ± 0.79	17.27
2k	Et	89.31 ± 4.39	0.13 ± 0.03	684.40
2l	Et	22.61 ± 3.11	0.03 ± 0.00	916.56
2m	Me	31.75 ± 0.46	0.55 ± 0.02	57.99
2n	Me	15.70 ± 1.41	0.08 ± 0.00	187.32
2o	Et	62.09 ± 10.20	0.06 ± 0.01	1 014.93
2p	Me	74.80 ± 9.31	0.66 ± 0.18	112.94
2q	Et	66.34 ± 4.12	0.12 ± 0.01	550.22
2r	Me	45.15 ± 4.28	0.11 ± 0.06	401.64
2s	Et	55.80 ± 1.46	0.02 ± 0.01	3 401.77
2t	Me	18.77 ± 0.42	0.23 ± 0.02	80.04
2u	Et	35.03 ± 10.07	0.05 ± 0.01	663.13
2v	Me	24.74 ± 1.18	0.27 ± 0.02	91.20
2w	Et	59.62 ± 0.79	0.01 ± 0.01	4 881.85
2x	Et	44.30 ± 8.79	0.13 ± 0.07	333.42
DB02		> 200.00	0.03 ± 0.01	> 7 750.81
3TC		> 200.00	0.61 ± 0.72	> 325.42

非核苷类逆转录酶抑制剂DB02氨基酸衍生物的合成及抗HIV-1活性研究

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杨金轩 ¹^,³^,^# , 余乐 ²^,^# , 杨玉卓 ² , 罗荣华 ¹ , 何严萍 ²^,^* , 郑永唐 ¹^,^*

药学学报 | 研究论文 2023,58(2): 405-412

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药学学报 | 研究论文 2023, 58(2): 405-412

非核苷类逆转录酶抑制剂DB02氨基酸衍生物的合成及抗HIV-1活性研究

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杨金轩¹^,³^,^#, 余乐²^,^#, 杨玉卓², 罗荣华¹, 何严萍²^,^*, 郑永唐¹^,^*

作者信息

1.中国科学院昆明动物研究所, 中国科学院动物模型与人类疾病机理重点实验室, 云南昆明 650223

2.云南大学化学科学与工程学院, 自然资源药物化学教育部重点实验室, 云南昆明 650091

3.云南中医药大学中药学院, 云南昆明 650500

通讯作者:

*何严萍, Tel/Fax: 86-871-65031119, E-mail: yphe@ynu.edu.cn;

郑永唐, Tel/Fax: 86-871-65195684, E-mail: zhengyt@mail.kiz.ac.cn

Design, synthesis and biological activity of DB02 amino acid derivatives as HIV-1 non-nucleoside reverse transcriptase inhibitors

Jin-xuan YANG¹^,³, Le YU², Yu-zhuo YANG², Rong-hua LUO¹, Yan-ping HE²^,^*, Yong-tang ZHENG¹^,^*

Affiliations

1. Key Laboratory of Animal Models and Human Disease Mechanisms of the Chinese Academy of Sciences, Kunming Institute of Zoology, Chinese Academy of Sciences, Kunming 650223, China

2. Key Laboratory of Medicinal Chemistry for Natural Resource Ministry of Education, School of Chemical Science and Technology, Yunnan University, Kunming 650091, China

3. College of Traditional Chinese Medicine, Yunnan University of Chinese Medicine, Kunming 650500, China

出版时间: 2023-02-12 doi: 10.16438/j.0513-4870.2022-0739

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摘要

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为提高非核苷类HIV-1逆转录酶抑制剂DB02氨基酸酯衍生物的稳定性, 本文基于生物电子等排原理, 以具有更高化学稳定性的酰胺替代酯键, 设计合成了24个DB02氨基酸酰胺衍生物2a~2x。采用MTT法及合胞体计数评估了其体外抗HIV-1活性。研究发现大部分目标化合物具有良好的抗HIV-1活性, 其中活性最佳的5个化合物2d、2i、2l、2s、2w的抗病毒效果均优于先导化合物DB02, 且具有优良的治疗指数(TI > 1 000.00)。这类化合物的构效关系研究为DB02衍生物的进一步开发提供了新的思路。

关键词

DB02 / 非核苷类逆转录酶抑制剂 / 氨基酸酰胺衍生物 / 抗HIV-1活性 / 构效关系

Abstract

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To improve the stability of amino acid ester derivatives of DB02, a series of 24 amide derivatives of DB02 amino acids as non-nucleoside HIV-1 reverse transcriptase inhibitor were designed and synthesized based on bioisosterism by replacing amino acid ester scaffold with more stable amide bond. The anti-HIV-1 activity of these compounds was evaluated by MTT assay and counting the number of syncytia. Most of the target compounds showed a potential anti-HIV-1 activity, among which compounds 2d, 2i, 2l, 2s, and 2w had better antiviral effect than lead compound DB02, with a therapeutic index > 1 000.00. Finally, the structure-activity relationship of these compounds was discussed, which provided new ideas for the further development of DB02 derivatives.

Key words

DB02 / non-nucleoside reverse transcriptase inhibitor / amino acid amide derivative / anti-HIV-1 activity / structure-activity relationship

引用本文

杨金轩, 余乐, 杨玉卓, 罗荣华, 何严萍, 郑永唐. 非核苷类逆转录酶抑制剂DB02氨基酸衍生物的合成及抗HIV-1活性研究. 药学学报, 2023 , 58 (2) : 405 -412 . DOI: 10.16438/j.0513-4870.2022-0739

Jin-xuan YANG, Le YU, Yu-zhuo YANG, Rong-hua LUO, Yan-ping HE, Yong-tang ZHENG. Design, synthesis and biological activity of DB02 amino acid derivatives as HIV-1 non-nucleoside reverse transcriptase inhibitors[J]. Acta Pharmaceutica Sinica, 2023 , 58 (2) : 405 -412 . DOI: 10.16438/j.0513-4870.2022-0739

正文

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获得性免疫缺陷综合征(acquired immune deficiency syndrome, AIDS) 是由人类免疫缺陷病毒(human immunodeficiency virus, HIV) 引起的一种致命性传染病。截至2020年全球大约有3 770万感染人群, 新增150万感染人群, 68万人死于HIV相关的疾病^[1]。目前HIV/AIDS治疗最为有效的方法是联合抗逆转录病毒疗法(combined antiretroviral therapy, cART), 一般是2种核苷类逆转录酶抑制剂+1种非核苷类逆转录酶抑制剂(non-nucleoside reverse transcriptase inhibitors, NNRTIs) 或整合酶链转移抑制剂或蛋白酶抑制剂^[2]。NNRTIs因其强有力的抗病毒活性、高选择性、适度的毒性和良好的药代动力学特性, 在cART治疗中独具优势, 也是药物研发的热点^[3]。尽管当前已有奈韦拉平、地拉韦啶、依法韦仑、依曲韦林、利匹韦林、多拉韦林、依磺韦林和艾诺维林等8种NNRTIs获批应用于临床, 但由于临床治疗方案的选择、药物间相互作用、药代性质不理想、毒副作用及耐药株的产生等问题^[4-9], 迫切需要不断开发新的抗HIV药物及先导化合物。

本课题组前期报道了系列二氢芳基/烷基硫-环己甲基嘧啶酮类(dihydro-aryl/alkylsulfanyl-cyclohexylmethyl-oxopyrimidines, S-DACOs) NNRTIs^[10], 其中优选化合物DB02对不同亚型不同嗜性的HIV-1病毒株均有较好的抑制作用(EC₅₀为2.40~41.80 nmol·L^-1), 且细胞毒性低(CC₅₀ > 1.00 mmol·L^-1)^[11]。但由于分子结构的疏水性, DB02水溶性极差, 导致生物利用度较低^[12], 阻碍了其进一步开发。为提高该类化合物的生物利用度, 在DB02的C-2侧链末端苯环通过酯键引入各种氨基酸, 进一步合成高效低毒的DB02-氨基酸酯衍生物1, 并利用其末端氨基制备相应的盐酸盐, 从而显著提高了其水溶性^[13]。遗憾的是, 在进一步的研究中发现化合物1由于氨基的碱性, 容易导致酯键断裂、分解生成化合物1b (图 1), 影响了其化学稳定性。

为分析化合物1与HIV-1逆转录酶(reversetranscriptase, RT) 的作用机制及为进一步的结构修饰提供指导, 采用AutoDock 4.2程序进行分子模拟^{[14, 15]}建立了优选化合物1a与RT (PDB: 1RT) 复合物模型。如图 2A所示, 化合物1a结合位点与DB02相似^[11], 其嘧啶环的N₃-NH与Lys101通过氢键作用将抑制剂分子锚定在RT内, C-6位的环己基以椅式柔性构象嵌入Trp229、Tyr181、Tyr188所组成的疏水口袋; C-2位β-羰基氧与Tyr318通过形成氢键对C-2位侧链起定向作用, 使其末端朝向溶剂开口处。与DB02结合位点不同的是, 化合物1a的C-2侧链酯羰基及末端氨基与Val106形成了双重氢键, 从而加强了小分子抑制剂与RT的结合, 这是其活性较DB02增强的重要原因。基于该复合物模型, 本研究用化学稳定性更好的酰胺键替代酯键设计了目标分子2 (图 2B), 酰胺键作为酯键的生物电子等排体, 同样可与Val106形成双重氢键, 因而有利于活性。另外, 由于C-2侧链末端处于溶剂开口处, 周围有较大空间可容纳不同取代基, 因此, 在该位置引入不同体积和构型的取代基(R²) 以寻找更优活性基团。期望化合物2在增加分子稳定性的同时保持其良好的抗HIV活性。

结果与讨论

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1 化学实验

以9-芴基甲氧基羰基(Fmoc) 保护的氨基酸3为起始原料, 与二氯亚砜反应生成N-Fmoc-氨基酸酰氯4, 再与对氨基苯乙酮5反应生成中间体6, 然后经溴化铜溴代得到中间体7。中间体7与5-烷基-2-硫嘧啶酮8在K₂CO₃催化下经烷基化反应生成氨基酸酰胺衍生物9。最后, 9在哌啶和二氯甲烷作用下脱保护得到目标化合物2。化合物的合成见合成路线1。

通过上述方法合成了24个目标新化合物, 并采用¹H NMR、¹³C NMR和HRMS-ESI进行了结构表征, 其理化性质和谱学数据见表 1和表 2。

DB02-氨基酸酯衍生物1在冰箱内-2 ℃密封储存, TLC检测24 h后即有杂点产生, 表明化合物开始分解。本研究合成的目标化合物2同条件储存, 经TLC跟踪检测60天以上仍无杂点生成, 说明以酰胺键替代酯键后的化合物具有良好的稳定性。

2 抗HIV-1活性评价

以临床使用的NNRTI拉米夫定(lamivudine, 3TC) 为阳性对照, 采用MTT法及合胞体计数评估了目标化合物2a~2x的体外抗病毒活性(表 3)。由表 3可知, 目标化合物普遍具有良好的抗HIV-1活性, 其EC₅₀介于0.01~2.59 µmol·L^-1之间。除化合物2a、2e、2g、2j和2p外, 其余化合物的EC₅₀值均低于阳性对照药物3TC (EC₅₀ = 0.61 µmol·L^-1), 其中2d、2i、2l、2s、2w这5个化合物的EC₅₀值均优于DB02 (EC₅₀ = 0.03 µmol·L^-1)。此外, 大部分目标化合物的细胞毒性较DB02明显增加, 除化合物2a、2b和2f的细胞毒性较小(CC₅₀ > 200.00 µmol·L^-1), 其余化合物的CC₅₀均介于15.00~150.60 µmol·L^-1。体外抗HIV-1活性研究显示化合物2b、2d、2i、2o、2s和2w的抗病毒效果最好, TI值均大于1 000.00, 其中活性最好的化合物2w的TI值高达4 881.85, 较阳性对照药物3TC提高了15倍。

此外, 由表 3可见, 该类目标化合物中嘧啶环C-5位取代基对活性有着明显影响, 5-乙基衍生物活性显著优于5-甲基衍生物, 5-乙基衍生物2b、2d、2i、2k、2l、2s、2w较5-甲基衍生物2a、2c、2h、2j、2m、2r、2v的抗HIV-1活性提高了4~31倍。就引入氨基酸而言, 除甘氨酸(2a) 及氨基己酸(2j) 的引入使活性略低外, 其余氨基酸衍生物无论其侧链基团为直链、支链或环均具有较好的抗病毒活性。另外, 氨基酸的构型对活性也有影响, 引入L-脯氨酸、L-亮氨酸和L-缬氨酸的化合物2w、2s、2l其活性要优于相应的D-构型衍生物2x、2q、2o, 当引入丙氨酸时, 则L-构型衍生物(2e、2f) 较其D-构型衍生物(2c、2d) 的活性降低。结合上述分子模型分析, 该系列化合物C-2侧链末端已进入溶剂开口处, 周围空间较大, 该位置除酰胺键羰基及末端氨基与V106的双重氢键作用外, 其余取代基对小分子抑制剂与RT结合的贡献较小。因此, 尽管取代基R²结构各异, 但其对活性的影响没有表现出明显的规律。今后研究将尝试引入更多类型氨基酸, 对其构效关系进行深入探讨。

3 小结

为提高NNRTIs-DB02的成药性, 本文设计并合成了24个DB02氨基酸酰胺衍生物(2a~2x), 并采用¹H NMR和¹³C NMR进行了结构确证。体外细胞水平抗HIV-1活性研究表明, 目标化合物普遍具有良好的抗HIV-1活性(EC₅₀ = 0.01~2.59 µmol·L^-1), 大部分化合物优于阳性对照药物3TC, 其中化合物2d、2i、2l、2s、2w的EC₅₀值均优于先导化合物DB02且具有优良的治疗指数(TI > 1 000.00)。初步构效关系分析表明, 该类化合物嘧啶环C-5位取代基及C-2侧链末端苯环上引入氨基酸的种类和构型均对活性有明显影响。将进一步尝试引入更多类型的氨基酸并借助氨基酸成盐反应提高化合物水溶性, 开发具有良好药代动力性质的抗HIV候选药物。

实验部分

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市售分析纯(AR) 或试剂级(RG) 或是根据反应所需自行制备得到。常压柱色谱所用硅胶(100/200~300目) 购自青岛海洋化工。薄层色谱(TLC) 板购自乳山市太阳干燥剂有限公司。噻唑蓝(thiazolyl blue tetrazolium bromide, MTT)、青霉素(penicillin)、氯化钠、氯化钾、磷酸二氢钾、磷酸氢二钠、TWEEN 20为Sigma公司产品。十二烷基硫酸钠(sodium dodecyl sulfate, SDS) 为BioFR0XX公司产品。二甲基亚砜(dimethyl sulfoxide, DMSO) 为VWR Life Science AMRESCO公司产品。硫酸链霉素(streptomycin sulfate) 为Solarbio公司产品。N, N-二甲基甲酰胺(N, N-dimethylformamine, DMF) 为成都市科隆化学品有限公司产品。RPMI-1640、胎牛血清(fetal bovine serum, FBS) 为Gibco公司产品。拉米夫定(lamivudine, 3TC) 购自大连美仑生物技术有限公司。人T淋巴细胞C8166细胞系、实验株HIV-1_IIIB由NIH AIDS Reagent Progro惠赠。

Bruker DRX 400 (瑞士Bruker公司), 内标TMS, 溶剂DMSO-d₆; 数字熔点仪(WRS-1, 中国卓光科技优先公司); Agilent LC/MSD TOF质谱仪(美国Agilent公司); 离心机(Allegra X-30, 美国Beckman公司); 二级生物安全柜(1287)、CO₂细胞培养箱(3111)、-80 ℃冰箱(386L -86 ℃) (美国Thermo公司); 2~8 ℃冰箱(HYC-360)、-20 ℃冰箱(BCD-196E/D) (中国Haier公司); 酶标仪(800TS, 美国Bioteck公司); 倒置显微镜(TS-100F, 日本Nikon公司)。

1 化合物的合成

1.1 N-Fmoc-(2-(4-乙酰苯氨基)-2-氧代乙基)氨基甲酸酯的合成

取250 mL的干燥圆底烧瓶, 将N-Fmoc-氨基酸3 (10 mmol) 溶于150 mL二氯甲烷溶液中, 滴加0.75 mL二氯亚砜(10 mmol), 加热回流反应5 h后, 减压蒸馏除去溶剂, 真空干燥2 h, 得中间体N-Fmoc-氨基酸酰氯4, 用150 mL无水乙醚溶解中间体4, 快速加入氨基苯乙酮5 (10 mmol), 剧烈震荡5 min, 有固体生成, 抽滤, 滤饼用50 mL的水洗涤, 真空干燥48 h得化合物6, 收率为62%~89%。可不经纯化直接用于下一步反应。

1.2 (9H-芴-9-基)甲基-(((4-(2-溴乙酰基)苯基)氨基))氨基甲酸酯的合成

在三口烧瓶中加入上一步的固体产物6, 再加入4.467 g (约20 mmol) 溴化铜粉末、120 mL (乙酸乙酯-氯仿= 1∶1, V/V) 的混合溶剂, 加热回流20 min, 加入20 mL无水乙醇, 反应2 h后停止加热, 趁热抽滤, 滤饼用冷氯仿(30 mL) 洗涤。合并滤液, 用无水硫酸镁干燥, 减压蒸馏得淡黄色固体; 无水乙醇重结晶得化合物7, 该粗产物可直接投下一步。

1.3 目标化合物的合成

在50 mL干燥圆底烧瓶中加入2 mmol的5-烷基-2-硫嘧啶酮8和10 mL DMF, 待完全溶解后加入2 mmol无水碳酸钾, 常温搅拌0.5 h, 加入上一步所得中间体7 (2 mmol), TLC跟踪检测反应至原料点消失, 将反应液倒入冰水中, 剧烈搅拌, 产生黄色浑浊物, 用乙酸乙酯萃取3次, 合并有机层并用饱和食盐水洗涤3次, 无水硫酸钠干燥, 减压蒸馏除去溶剂, 得到黄色固体粗产品(9), 用6 mL二氯甲烷溶解(9), 再加入3 mL哌啶, 常温下反应30 min, 减压蒸馏除去溶剂。即获得油状物质, 用无水乙醚少量多次洗去哌啶, 干燥后经过重结晶纯化得到目标化合物(2)。收率: 30%~69%。

2 目标化合物的抗HIV-1活性测试

2.1 化合物对C8166细胞的毒性实验

96孔板内将药物进行5倍梯度稀释, 共设置6个药物浓度, 每个药物浓度设3个复孔(每孔100 μL), 同时设置无药物的阴性对照孔(NC) 和空白对照孔(BC)。除BC孔外, 每孔加入100 μL C8166细胞悬液(细胞密度为每毫升4×10⁵个)。在37 ℃、5% CO₂培养箱培养3天, 采用MTT法测定化合物对C8166细胞系的毒性。800TS酶标仪在570/630 nm波长下测定OD值, 计算半数细胞毒性浓度(the 50% cytotoxicity concentration, CC₅₀)^{[16, 17]}。

2.2 化合物对HIV-1_IIIB致C8166细胞病变的抑制作用

96孔板内将药物进行5倍梯度稀释, 共设置6个药物浓度, 每个药物浓度设3个复孔(每孔100 μL)。每孔加50 μL C8166细胞悬液(细胞密度为每毫升8×10⁵个) 与50 μL HIV-1_IIIB病毒稀释液(1300 TCID50), 同时设置含C8166细胞与病毒的阳性对照孔(PC), 置于37 ℃、5% CO₂培养箱内培养。第3天在倒置显微镜下(100×) 观察合胞体的形成并计数, 计算半数有效浓度(the 50% effective concentration, EC₅₀)^[18]。

作者贡献: 杨金轩负责化合物体外抗HIV-1活性测试及文章撰写; 余乐负责化合物的合成; 杨玉卓负责数据的整理及文章部分撰写; 罗荣华协助活性测试和文章的修改; 何严萍和郑永唐负责课题规划、实验指导及文章修改及校对。

利益冲突: 无利益冲突。

基金

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国家自然科学基金资助项目(82060670)
国家自然科学基金资助项目(U1702286)
国家自然科学基金资助项目(21967020)
云南省重大科技专项计划课题(202103AC100005)
中缅国际合作项目(2019YFE0109200)

参考文献

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2023年第58卷第2期

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文章信息

doi: 10.16438/j.0513-4870.2022-0739

接收时间：2022-06-16
首发时间：2025-11-21
出版时间：2023-02-12

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收稿日期：2022-06-16
修回日期：2022-07-18

基金

国家自然科学基金资助项目(82060670)

国家自然科学基金资助项目(U1702286)

国家自然科学基金资助项目(21967020)

云南省重大科技专项计划课题(202103AC100005)

中缅国际合作项目(2019YFE0109200)

作者信息

1.中国科学院昆明动物研究所, 中国科学院动物模型与人类疾病机理重点实验室, 云南昆明 650223

2.云南大学化学科学与工程学院, 自然资源药物化学教育部重点实验室, 云南昆明 650091

3.云南中医药大学中药学院, 云南昆明 650500

通讯作者:

*何严萍, Tel/Fax: 86-871-65031119, E-mail: yphe@ynu.edu.cn;

郑永唐, Tel/Fax: 86-871-65195684, E-mail: zhengyt@mail.kiz.ac.cn

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https://castjournals.cast.org.cn/joweb/yxxb/CN/10.16438/j.0513-4870.2022-0739

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2种不同金属材料的力学参数

科 Family	属数 Number of genus	种数 Number of species	占总种数比例 Percentage of total species (%)	属 Genus	种数 Number of species	占总种数比例 Percentage of total species (%)
鹅膏菌科Amanitaceae	2	11	5.26	鹅膏菌属 Amanita	10	4.78
小菇科 Mycenaceae	2	12	5.74	丝盖伞属 Inocybe	5	2.39
多孔菌科 Polyporaceae	8	14	6.70	蜡蘑属 Laccaria	5	2.39
红菇科 Russulaceae	3	23	11.00	小皮伞属 Marasmius	6	2.87
				小菇属 Mycena	11	5.26
				光柄菇属 Pluteus	5	2.39
				红菇属 Russula	17	8.13
				栓菌属 Trametes	5	2.39

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Compd.	Yield/%	mp/℃		Compd.	Yield/%	mp/℃
2a	55	160.8-161.8		2m	62	148.1-149.1
2b	45	157.3-158.3	2n	58	111.7-112.7
2c	41	172.1-172.1	2o	55	106.1-107.1
2d	53	174.2-175.2	2p	44	138.9-139.9
2e	40	174.7-175.7	2q	39	141.3-142.3
2f	49	176.3-177.3	2r	43	124.8-125.8
2g	58	199.6-200.6	2s	56	141.6-142.6
2h	53	142.2-143.2	2t	62	115.0-116.0
2i	49	159.8-160.8	2u	54	117.7-118.7
2j	40	197.1-198.1	2v	64	136.2-137.2
2k	48	203.7-204.7	2w	61	139.3-149.3
2l	52	162.5-163.5	2x	61	131.5-132.5

Compd.

Yield/%

mp/℃

Compd.

Yield/%

mp/℃

160.8-161.8

148.1-149.1

157.3-158.3

111.7-112.7

172.1-172.1

106.1-107.1

174.2-175.2

138.9-139.9

174.7-175.7

141.3-142.3

176.3-177.3

124.8-125.8

199.6-200.6

141.6-142.6

142.2-143.2

115.0-116.0

159.8-160.8

117.7-118.7

197.1-198.1

136.2-137.2

203.7-204.7

139.3-149.3

162.5-163.5

131.5-132.5

Compd.	¹H NMR, ¹³C NMR, HRMS-ESI
2a	¹H NMR (400 MHz, DMSO-d₆) δ: 8.02 (d, J = 8.5 Hz, 2H, ArH), 7.86 (d, J = 9.2 Hz, 2H, ArH), 4.60 (s, 2H, CH₂-S), 3.61 (d, J = 2.9 Hz, 2H, NH₂CH₂), 2.09 (d, J = 6.7 Hz, 2H, CH₂-cyclohexyl), 1.79 (s, 3H, Me), 1.66 (m, 7H, Cyclohexyl), 1.54 (m, 4H, cyclohexyl); ¹³C NMR (100 MHz, DMSO-d₆) δ: 190.3, 174.3, 161.5 (2C), 155.9, 142.8, 131.1, 130.0 (2C), 118.7 (2C), 115.3, 43.9, 37.6 (2C), 36.8 (2C), 32.9, 32.6 (3C), 10.9; HRMS-ESI: m/z calcd. for C₂₂H₂₈N₄O₃S [M+H]⁺ 429.194 6, found 429.194 8.
2b	¹H NMR (400 MHz, DMSO-d₆) δ: 8.03 (d, J = 8.5 Hz, 2H, ArH), 7.83 (d, J = 9.9 Hz, 2H, ArH), 4.60 (s, 2H, CH₂-S), 3.60 (d, J = 2.9 Hz, 2H, NH₂CH₂), 2.29-2.26 (m, 2H, CH₂CH₃), 2.08 (d, J = 6.9 Hz, 2H, CH₂-cyclohexyl), 1.65 (m, 7H, cyclohexyl), 1.53 (dd, J = 6.1, 2.1 Hz, 4H, cyclohexyl), 0.91 (s, 3H, CH₂CH₃); ¹³C NMR (100 MHz, DMSO-d₆) δ: 192.3, 174.4, 161.7 (2C), 156.4, 143.2, 131.0, 130.0 (2C), 118.7 (2C), 117.2, 43.9, 37.6, 36.6 (2C), 32.9 (2C), 32.6 (3C), 22.5, 13.7; HRMS-ESI: m/z calcd. for C₂₃H₃₀N₄O₃S [M+H]⁺ 443.210 3, found 443.210 5.
2c	¹H NMR (400 MHz, DMSO-d₆) δ: 8.02 (s, 2H, ArH), 7.88-7.79 (m, 2H, ArH), 4.63-4.55 (m, 2H, CH₂-S), 3.62-3.55 (m, 1H, CH-NH₂), 2.15-2.04 (m, 2H, CH₂-cyclohexyl), 1.83-1.77 (m, 3H, Me), 1.68-1.60 (m, 1H, cyclohexyl), 1.54-1.48 (m, 3H, CHCH₃), 1.35-1.21 (m, 7H, cyclohexyl), 1.02-0.90 (m, 3H, cyclohexyl); ¹³C NMR (100 MHz, DMSO-d₆) δ: 191.9, 174.7, 164.1, 160.9, 156.4, 143.4, 131.0, 129.5 (2C), 118.3 (2C), 114.7, 50.87, 36.6, 36.3, 32.4 (2C), 25.8, 25.5 (3C), 20.7, 10.4; HRMS-ESI: m/z calcd. for C₂₃H₃₀N₄O₃S [M+H]⁺ 443.210 3, found 443.210 6.
2d	¹H NMR (400 MHz, DMSO-d₆) δ: 8.02 (d, J = 8.4 Hz, 2H, ArH), 7.82 (d, J = 8.5 Hz, 2H, ArH), 4.60 (s, 2H, CH₂-S), 3.54 (q, J = 7.1 Hz, 1H, CH-NH₂), 2.28 (q, J = 7.4 Hz, 2H, CH₂CH₃), 2.08 (d, J = 6.7 Hz, 2H, CH₂-cyclohexyl), 1.50-1.40 (m, 3H, CHCH₃), 1.36-1.28 (m, 2H, cyclohexyl), 1.25 (t, J = 6.1 Hz, 5H, cyclohexyl), 0.91 (t, J = 7.2 Hz, 7H, cyclohexyl and CH₂CH₃); ¹³C NMR (100 MHz, DMSO-d₆) δ: 191.9, 174.9, 163.2, 160.44, 156.32, 143.51, 130.96, 129.55 (2C), 118.33 (2C), 114.63, 50.95, 36.76, 36.14, 32.50 (2C), 25.83, 25.55 (3C), 20.87, 17.97, 13.27; HRMS-ESI: m/z calcd. for C₂₄H₃₂N₄O₃S [M+H]⁺ 457.225 9, found 457.226 1.
2e	¹H NMR (400 MHz, DMSO-d₆) δ: 8.07 (s, 2H, ArH), 7.86-7.72 (m, 2H, ArH), 4.60 (s, 2H, CH₂-S), 3.72 (m, 1H, CH-NH₂), 2.15-2.01 (m, 2H, CH₂-cyclohexyl), 1.80-1.74 (s, 3H, Me), 1.71-1.62 (m, 1H, cyclohexyl), 1.52-1.44 (m, 3H, CHCH₃), 1.36-1.09 (m, 7H, cyclohexyl), 1.03-0.90 (m, 3H, cyclohexyl); ¹³C NMR (100 MHz, DMSO-d₆) δ: 191.6, 174.7, 163.2, 160.5, 154.1, 144.0, 131.8, 129.0 (2C), 117.8 (2C), 114.0, 51.0, 36.8, 36.3, 32.8 (2C), 25.9, 25.3 (3C), 18.8, 10.3; HRMS-ESI: m/z calcd. for C₂₃H₃₀N₄O₃S [M+H]⁺ 443.210 3, found 443.210 7.
2f	¹H NMR (400 MHz, DMSO-d₆) δ: 8.04 (d, J = 8.4 Hz, 2H, ArH), 7.80 (d, J = 8.5 Hz, 2H, ArH), 4.61 (s, 2H, CH₂-S), 3.60 (q, J = 7.8 Hz, 1H, CH-NH₂), 2.18 (q, J = 7.2 Hz, 2H, CH₂CH₃), 2.08 (d, J = 7.0 Hz, 2H, CH₂-cyclohexyl), 1.56-1.40 (m, 3H, CHCH₃), 1.35-1.26 (m, 2H, cyclohexyl), 1.21 (t, J = 6.1 Hz, 5H, cyclohexyl), 0.91 (t, J = 7.2 Hz, 7H, cyclohexyl and CH₂CH₃); ¹³C NMR (100 MHz, DMSO-d₆) δ: 191.2, 174.6, 163.8, 161.1, 156.3, 144.7, 131.0, 129.5 (2C), 118.1 (2C), 114.3, 51.8, 36.9 (2C), 35.4 (2C), 31.6, 26.6 (2C), 25.1, 20.2, 18.3, 14.1; HRMS-ESI: m/z calcd. for C₂₄H₃₂N₄O₃S [M+H]⁺ 457.225 9, found 457.226 3.
2g	¹H NMR (400 MHz, DMSO-d₆) δ: 8.09 (d, J = 8.4 Hz, 2H, ArH), 7.82 (d, J = 8.5 Hz, 2H, ArH), 4.68 (s, 2H, CH₂-S), 3.11 (m, 2H, NH₂CH₂CH₂), 2.60 (m, 2H, NH₂CH₂CH₂), 2.28 (m, 2H, CH₂CH₃), 2.08 (d, J = 7.8 Hz, 2H, CH₂-cyclohexyl), 1.56-1.29 (m, 11H, cyclohexyl), 0.91 (t, J = 7.2 Hz, 3H, CH₂CH₃); ¹³C NMR (100 MHz, DMSO-d₆) δ: 191.6, 173.6, 162.1, 160.8, 157.9, 143.6, 131.0, 129.6 (2C), 119.2 (2C), 114.3, 39.8, 36.9, 36.0, 34.5, 33.8 (2C), 32.7, 26.4, 25.3 (2C), 20.2, 14.2; HRMS-ESI: m/z calcd. for C₂₄H₃₂N₄O₃S [M+H]⁺ 457.225 9, found 457.226 0.
2h	¹H NMR (400 MHz, DMSO-d₆) δ: 8.02 (d, J = 8.5 Hz, 2H, ArH), 7.82 (d, J = 8.8 Hz, 2H, ArH), 4.58 (s, 2H, CH₂-S), 3.40 (t, J = 6.6 Hz, 1H, CH-NH₂), 2.07 (s, 3H, Me), 1.79 (m, 4H, CH₂-cyclohexyl and CH₂CH₂CH₂CH₃), 1.66-0.91 (m, 15H, cyclohexyl and CH₂CH₂CH₂CH₃), 0.84 (t, J = 6.9 Hz, 3H, CH₂CH₂CH₂CH₃); ¹³C NMR (100 MHz, DMSO-d₆) δ: 192.0, 174.7, 163.7, 161.0, 156.2, 143.5, 131.0, 129.5 (2C), 118.3 (2C), 114.9, 55.4, 36.6, 36.2, 34.4, 32.4 (2C), 27.3, 25.8 (3C), 25.5, 22.1, 13.9, 10.4; HRMS-ESI: m/z calcd. for C₂₆H₃₆N₄O₃S [M+H]⁺ 485.257 2, found 485.256 8.
2i	¹H NMR (400 MHz, DMSO-d₆) δ: 8.06 (d, J = 8.4 Hz, 2H, ArH), 7.84 (d, J = 8.7 Hz, 2H, ArH), 4.59 (s, 2H, CH₂-S), 3.37 (t, J = 6.5 Hz, 1H, CH-NH₂), 2.28 (q, J = 7.3 Hz, 2H, CH₂CH₃), 2.06 (d, J = 6.7 Hz, 2H, CH₂-cyclohexyl), 1.69-1.17 (m, 13H, cyclohexyl and CH₂CH₂CH₂CH₃), 0.88 (dt, J = 25.0, 7.2 Hz, 10H, CH₂CH₃ and CH₂CH₂CH₂CH₃); ¹³C NMR (100 MHz, DMSO-d₆) δ: 191.9, 174.8, 163.1 (2C), 156.2, 143.5, 130.9, 129.5 (2C), 120.9 (2C), 118.3, 55.5, 36.7, 36.1, 34.5, 32.4 (2C), 27.4, 25.8, 25.5 (2C), 22.1, 17.9, 13.8, 13.2; HRMS-ESI: m/z calcd. for C₂₇H₃₈N₄O₃S [M+H]⁺ 499.272 9, found 499.272 6.
2j	¹H NMR (400 MHz, DMSO-d₆) δ: 8.02 (d, J = 8.4 Hz, 2H, ArH), 7.82 (d, J = 8.8 Hz, 2H, ArH), 4.63 (s, 1H, CH₂-S), 2.78 (t, J = 7.6 Hz, 2H, NH₂CH₂CH₂CH₂CH₂CH₂), 2.40 (t, J = 7.3 Hz, 2H, NH₂CH₂CH₂CH₂CH₂CH₂), 2.28 (d, J = 7.6 Hz, 2H, CH₂-cyclohexyl), 2.08 (s, 3H, Me), 1.66 (q, J = 5.8 Hz, 4H, NH₂CH₂CH₂CH₂CH₂CH₂), 1.61-1.57 (m, 4H, cyclohexyl and NH₂CH₂CH₂CH₂CH₂CH₂), 1.57-1.50 (m, 4H, cyclohexyl), 1.37-1.25 (m, 5H, cyclohexyl); ¹³C NMR (100 MHz, DMSO-d₆) δ: 192.1, 175.5, 164.1 (2C), 155.3, 144.2, 131.0, 129.8 (2C), 118.0 (2C), 117.0, 43, 0, 37.1, 32.9 (2C), 27.2, 26.4 (2C), 25.7 (3C), 24.6, 13.7; HRMS-ESI: m/z calcd. for C₂₆H₃₆N₄O₃S [M+H]⁺ 485.257 2, found 485.257 7.
2k	¹H NMR (400 MHz, DMSO-d₆) δ: 8.00 (d, J = 8.5 Hz, 2H, ArH), 7.82 (d, J = 8.6 Hz, 2H, ArH), 4.60 (s, 2H, CH₂-S), 2.76 (t, J = 7.6 Hz, 2H, NH₂CH₂CH₂CH₂CH₂CH₂), 2.40 (t, J = 7.3 Hz, 2H, NH₂CH₂CH₂CH₂CH₂CH₂), 2.30 (m, 2H, CH₂CH₃), 2.08 (d, J = 6.8 Hz, 2H, CH₂-cyclohexyl), 1.66 (q, J = 5.8 Hz, 4H, NH₂CH₂CH₂CH₂CH₂CH₂), 1.61-1.57 (m, 4H, cyclohexyl), 1.57-1.50 (m, 4H, cyclohexyl and NH₂CH₂CH₂CH₂CH₂CH₂), 1.37-1.25 (m, 5H, cyclohexyl), 0.92 (m, 3H, CH₂CH₃); ¹³C NMR (100 MHz, DMSO-d₆) δ: 192.3, 175.8, 164.3 (2C), 155.2, 144.5, 131.1, 129.9 (2C), 118.6 (2C), 117.2, 42.8, 37.2 (2C), 32.9 (2C), 27.2, 26.2 (2C), 25.9 (2C), 24.9 (2C), 18.4, 13.7; HRMS-ESI: m/z calcd. for C₂₇H₃₈N₄O₃S [M+H]⁺ 499.272 9, found 499.273 0.
2l	¹H NMR (400 MHz, DMSO-d₆) δ: 8.03 (d, J = 8.4 Hz, 2H, ArH), 7.84 (d, J = 8.6 Hz, 2H, ArH), 4.60 (s, 2H, CH₂-S), 3.21 (d, J = 5.5 Hz, 1H, CH-NH₂), 2.28 (q, J = 7.4 Hz, 2H, CH₂CH₃), 2.07 (d, J = 6.8 Hz, 2H, CH₂-cyclohexyl), 1.94 (dq, J = 13.4, 6.8 Hz, 1H, CH(CH₃)₂), 1.49-1.39 (m, 6H, CH(CH₃)₂), 1.30-1.20 (m, 3H, CH₂CH₃), 0.89 (m, 11H, cyclohexyl); ¹³C NMR (100 MHz, DMSO-d₆) δ: 191.8, 174.3, 163.0, 160.8, 156.1, 143.3, 130.8, 129.4 (2C), 120.8 (2C), 118.2, 60.6, 36.6, 35.9, 32.3 (2C), 31.7, 30.1, 25.7, 25.4 (2C), 19.3, 17.8, 17.2, 13.2; HRMS-ESI: m/z calcd. for C₂₆H₃₆N₄O₃S [M+H]⁺ 485.257 2, found 485.257 5.
2m	¹H NMR (400 MHz, DMSO-d₆) δ: 8.03 (d, J = 8.4 Hz, 2H, ArH), 7.83 (d, J = 8.5 Hz, 2H, ArH), 4.59 (s, 2H, CH₂-S), 3.25 (d, J = 5.6 Hz, 1H, CH-NH₂), 2.07 (d, J = 6.7 Hz, 2H, CH₂-cyclohexyl), 1.96 (m, 1H, CH(CH₃)₂), 1.80 (s, 3H, Me), 1.61 (d, J = 21.3 Hz, 1H, cyclohexyl), 1.45 (d, J = 10.2 Hz, 4H, cyclohexyl), 1.32-1.19 (m, 4H, cyclohexyl), 1.05 (t, J = 7.0 Hz, 2H, cyclohexyl), 0.93 (d, J = 6.7 Hz, 3H, CH(CH₃)₂), 0.86 (d, J = 6.7 Hz, 3H, CH(CH₃)₂); ¹³C NMR (100 MHz, DMSO-d₆) δ: 192.0, 174.0, 163.6, 161.0, 156.0, 143.4, 131.0, 129.5 (2C), 120.5 (2C), 118.3, 60.5, 36.6, 36.2 (2C), 32.4 (2C), 31.7, 25.8, 25.5 (2C), 18.5 (2C), 10.42; HRMS-ESI: m/z calcd. for C₂₅H₃₄N₄O₃S [M+H]⁺ 471.241 6, found 471.242 0.
2n	¹H NMR (400 MHz, DMSO-d₆) δ: 8.03 (d, J = 8.9 Hz, 2H, ArH), 7.89-7.79 (m, 2H, ArH), 4.58 (s, 2H, CH₂-S), 3.27-3.20 (m, 1H, CH-NH₂), 2.06 (q, J = 5.7 Hz, 2H, CH₂-cyclohexyl), 1.95 (m, 1H, CH(CH₃)₂), 1.83-1.76 (m, 3H, Me), 1.52-1.18 (m, 6H, CH(CH₃)₂), 1.04-0.79 (m, 11H, cyclohexyl); ¹³C NMR (100 MHz, DMSO-d₆) δ: 192.0, 174.1, 163.6 (2C), 156.1, 143.4, 130.9, 129.5 (2C), 118.3 (2C), 114.8, 60.6, 36.6, 36.2, 32.4 (2C), 31.7 (2C), 25.8, 25.5 (2C), 19.4 (2C), 10.4; HRMS-ESI: m/z calcd. for C₂₅H₃₄N₄O₃S [M+H]⁺ 471.241 6, found 471.241 8.
2o	¹H NMR (400 MHz, DMSO-d₆) δ: 8.02 (d, J = 8.4 Hz, 2H, ArH), 7.83 (d, J = 8.8 Hz, 2H, ArH), 4.58 (s, 2H, CH₂-S), 3.19 (d, J = 5.5 Hz, 1H, CH-NH₂), 2.28 (q, J = 7.3 Hz, 2H, CH₂CH₃), 2.07 (d, J = 6.8 Hz, 2H, CH₂-cyclohexyl), 1.98-1.88 (m, 1H, CH(CH₃)₂), 1.65-1.19 (m, 9H, CH(CH₃)₂ and CH₂CH₃), 0.88 (m, 11H, cyclohexyl); ¹³C NMR (100 MHz, DMSO-d₆) δ: 192.0, 174.5, 163.6 (2C), 156.1, 143.5, 130.9, 129.5 (2C), 118.3 (2C), 114.6, 60.8, 36.6, 36.1, 32.5 (2C), 31.8 (2C), 25.8, 25.5 (2C), 19.4, 17.9, 17.2, 13.3; HRMS-ESI: m/z calcd. for C₂₆H₃₆N₄O₃S [M+H]⁺ 485.257 2, found 485.257 0.
2p	¹H NMR (400 MHz, DMSO-d₆) δ: 8.01 (d, J = 8.7 Hz, 2H, ArH), 7.83 (d, J = 8.2 Hz, 2H, ArH), 4.57 (s, 2H, CH₂-S), 3.43-3.40 (m, 1H, CH-NH₂), 2.08 (d, J = 6.7 Hz, 2H, CH₂-cyclohexyl), 1.77-1.70 (m, 1H, CH₂CH(CH₃)₂), 1.61 (m, 8H, Me and cyclohexyl), 1.54-1.51 (m, 5H, cyclohexyl and CH₂CH(CH₃)₂), 1.17 (m, 3H, cyclohexyl), 0.91 (m, 6H, CH₂CH(CH₃)₂); ¹³C NMR (100 MHz, DMSO-d₆) δ: 192.2. 175.8. 160.9. 144.0, 131.4, 131.2, 129.9 (2C), 118.8 (2C), 112.9, 54.5, 44.4, 36.7 (2C), 33.1 (2C), 33.0, 26.3, 26.1 (2C), 23.2, 22.6 (2C), 13.8; HRMS-ESI: m/z calcd. for C₂₆H₃₆N₄O₃S [M+H]⁺ 485.257 2, found 485.257 5.
2q	¹H NMR (400 MHz, DMSO-d₆) δ: 8.06 (d, J = 8.7 Hz, 2H, ArH), 7.85 (t, J = 8.2 Hz, 2H, ArH), 4.59 (s, 2H, CH₂-S), 3.44-3.41 (m, 1H, CH-NH₂), 2.29 (m, 2H, CH₂CH₃), 2.08 (d, J = 6.7 Hz, 2H, CH₂-cyclohexyl), 1.77-1.70 (m, 1H, CH₂CH(CH₃)₂), 1.64-1.60 (m, 8H, cyclohexyl), 1.54-1.50 (m, 5H, cyclohexyl and CH₂CH(CH₃)₂), 1.17 (t, J = 7.1 Hz, 3H, CH₂CH₃), 0.92 (m, 6H, CH₂CH(CH₃)₂); ¹³C NMR (100 MHz, DMSO-d₆) δ: 192.2, 175.8, 160.9, 144.0, 131.4, 131.2 (2C), 129.9 (2C), 118.8, 112.9, 54.5, 44.4, 36.7 (2C), 33.1 (2C), 33.0, 26.3, 26.1 (2C), 23.2, 22.6 (2C), 18.5, 13.8; HRMS-ESI: m/z calc.d for C₂₇H₃₈N₄O₃S [M+H]⁺ 499.272 9, found 499.273 3.
2r	¹H NMR (400 MHz, DMSO-d₆) δ: 8.02 (d, J = 8.4 Hz, 2H, ArH), 7.83 (d, J = 8.7 Hz, 2H, ArH), 4.59 (s, 2H, CH₂-S), 3.44 (dd, J = 8.5, 5.8 Hz, 1H, CH-NH₂), 2.08 (d, J = 6.6 Hz, 2H, CH₂-cyclohexyl), 1.80 (s, 3H, Me), 1.77-1.69 (m, 1H, CH₂CH(CH₃)₂), 1.54-1.20 (m, 10H, cyclohexyl and CH₂CH(CH₃)₂), 0.89 (m, 9H, cyclohexyl and CH₂CH(CH₃)₂); ¹³C NMR (100 MHz, DMSO-d₆) δ: 192.5, 175.5, 163.3, 161.4, 156.1, 144.0, 130.9, 129.9 (2C), 120.5 (2C), 118.4, 54.2, 44.2, 37.1, 36.7, 32.8 (2C), 30.1, 25.9 (2C), 25.8, 23.5, 22.4 (2C), 13.8; HRMS-ESI: m/z calcd. for C₂₆H₃₆N₄O₃S [M+H]⁺ 485.257 2, found 485.257 6.
2s	¹H NMR (400 MHz, DMSO-d₆) δ: 8.06-7.99 (m, 2H, ArH), 7.82 (t, J = 6.2 Hz, 2H, ArH), 4.60 (d, J = 4.0 Hz, 2H, CH₂-S), 3.43-3.39 (m, 1H, CH-NH₂), 2.28 (m, 2H, CH₂CH₃), 2.08 (d, J = 5.5 Hz, 2H, CH₂-cyclohexyl), 1.73 (dp, J = 13.4, 6.6 Hz, 1H, CH₂CH(CH₃)₂), 1.51-1.23 (m, 9H, cyclohexyl and CH₂CH(CH₃)₂), 0.97-0.83 (m, 13H, cyclohexyl and CH₂CH(CH₃)₂) and CH₂CH₃); ¹³C NMR (100 MHz, DMSO-d₆) δ: 191.9, 175.3, 163.2, 160.5, 156.3, 143.6, 130.9, 129.6 (2C), 120.9 (2C), 118.4, 54.1, 43.9, 36.7, 36.1, 32.5 (2C), 25.8, 25.6 (2C), 24.2, 23.1 (2C), 21.9, 17.9, 13.3; HRMS-ESI: m/z calcd. for C₂₇H₃₈N₄O₃S [M+H]⁺ 499.272 9, found 499.272 7.
2t	¹H NMR (400 MHz, DMSO-d₆) δ: 8.02 (d, J = 8.4 Hz, 2H, ArH), 7.80 (d, J = 8.6 Hz, 2H, ArH), 7.27-7.16 (m, 5H, ArH), 4.61 (s, 2H, CH₂-S), 3.66 (m, 1H, CH-NH₂), 3.01 (dd, J = 13.4, 5.7 Hz, 1H, CH₂-Ph), 2.77 (dd, J = 13.4, 7.7 Hz, 1H, CH₂-Ph), 2.09 (d, J = 6.6 Hz, 2H, CH₂-cyclohexyl), 1.80 (s, 3H, Me), 1.26 (t, J = 14.8 Hz, 7H, cyclohexyl), 0.92 (h, J = 13.4, 12.3 Hz, 4H, cyclohexyl); ¹³C NMR (100 MHz, DMSO-d₆) δ: 192.3, 174.4, 163.9 (2C), 156.4, 143.9, 138.7, 131.4, 130.0, 129.7 (2C), 128.5 (2C), 126.6 (2C), 118.8 (2C), 115.4, 57.5, 37.1, 36.7 (2C), 32.9 (2C), 26.3, 26.0 (3C), 10.8; HRMS-ESI: m/z calcd. for C₂₉H₃₄N₄O₃S [M+H]⁺ 519.241 6, found 519.242 0.
2u	¹H NMR (400 MHz, DMSO-d₆) δ: 8.02 (d, J = 8.4 Hz, 2H, ArH), 7.81 (d, J = 8.4 Hz, 2H, ArH), 7.26-7.16 (m, 5H, ArH), 4.61 (s, 2H, CH₂-S), 3.69 (m, 1H, CH-NH₂), 3.02 (dd, J = 13.4, 5.7 Hz, 1H, CH₂-Ph), 2.77 (dd, J = 13.4, 7.7 Hz, 1H, CH₂-Ph), 2.29 (q, J = 7.3 Hz, 2H, CH₂CH₃), 2.08 (d, J = 6.8 Hz, 2H, CH₂-cyclohexyl), 1.52-1.18 (m, 7H, cyclohexyl), 0.91 (t, J = 7.6 Hz, 7H, cyclohexyl and CH₂CH₃); ¹³C NMR (100 MHz, DMSO-d₆) δ: 192.3, 174.4, 163.6, 160.9, 156.7, 143.9, 138.7, 131.4, 129.9 (2C), 129.8 (2C), 128.5 (2C), 126.6, 121.4 (2C), 118.9, 57.5, 37.2, 36.6 (2C), 32.9 (2C), 26.3, 26.0 (3C), 18.4, 13.7; HRMS-ESI: m/z calcd. for C₃₀H₃₆N₄O₃S [M+H]⁺ 533.257 2, found 533.257 0.
2v	¹H NMR (400 MHz, DMSO-d₆) δ: 10.44 (s, 1H, C=O-NH), 8.02 (d, J = 8.4 Hz, 2H, ArH), 7.85 (d, J = 8.5 Hz, 2H, ArH), 4.59 (s, 2H, CH₂-S), 3.80 (m, 1H, pyrrolidinyl), 2.92 (t, J = 6.6 Hz, 2H, pyrrolidinyl), 2.08 (d, J = 7.0 Hz, 2H, CH₂-cyclohexyl), 1.79 (s, 3H, Me), 1.73-1.18 (m, 11H, cyclohexyl), 0.91 (m, 4H, pyrrolidinyl); ¹³C NMR (100 MHz, DMSO-d₆) δ: 192.5, 174.0, 164.1, 161.4, 156.5, 143.5, 131.6, 130.0 (2C), 118.8 (2C), 115.4, 61.2, 47.1, 37.1, 36.7, 32.8 (2C), 30.9 (2C), 26.3, 26.1 (2C), 25.9, 10.9; HRMS-ESI: m/z calcd. for C₂₅H₃₂N₄O₃S [M+H]⁺ 469.225 9, found 469.226 3.
2w	¹H NMR (400 MHz, DMSO-d₆) δ: 10.69 (s, 1H, C=O-NH), 8.20-7.70 (m, 4H, ArH), 4.59 (s, 2H, CH₂-S), 3.90 (s, 1H, pyrrolidinyl), 2.95 (m, 2H, pyrrolidinyl), 2.16 (m, 4H, CH₂-cyclohexyl and CH₂CH₃), 1.72-1.10 (m, 12H, cyclohexyl and pyrrolidinyl), 1.01-0.75 (m, 6H, pyrrolidinyl and CH₂CH₃); ¹³C NMR (100 MHz, DMSO-d₆) δ: 192.5, 173.2, 163.8, 160.8, 156.8, 143.5, 131.6, 129.9 (2C), 121.3 (2C), 118.8, 61.0, 46.9, 43.9, 37.2, 36.5, 32.9, 30.8, 26.3, 25.9, 25.8, 22.5, 22.2, 18.4, 13.7; HRMS-ESI: m/z calcd. for C₂₆H₃₄N₄O₃S [M+H]⁺ 483.241 6, found 483.241 8.
2x	¹H NMR (400 MHz, DMSO-d₆) δ: 10.69 (s, 1H, C=O-NH), 8.20 (d, J = 8.4 Hz, 2H, ArH), 7.80 (d, J = 8.6 Hz, 2H, ArH), 4.60 (s, 2H, CH₂-S), 3.90 (t, 1H, pyrrolidinyl), 2.95 (m, 2H, pyrrolidinyl), 2.16 (q, J = 8.5 Hz, 2H, CH₂CH₃), 2.06(d, J = 7.2, 2H, CH₂-cyclohexyl), 1.72-1.10 (m, 12H, cyclohexyl and pyrrolidinyl), 1.01-0.75 (m, 6H, pyrrolidinyl and CH₂CH₃); ¹³C NMR (100 MHz, DMSO-d₆) δ: 191.4, 172.2, 163.8, 159.8, 156.8, 146.2, 131.8, 129.9 (2C), 121.3 (2C), 118.8, 62.4, 46.9, 43.9, 37.2, 36.5, 32.9, 30.8, 26.3, 26.0, 25.8, 22.6, 22.3, 18.4, 14.0; HRMS-ESI: m/z calcd. for C₂₆H₃₄N₄O₃S [M+H]⁺ 483.241 6, found 483.242 0.

Compd.

¹H NMR, ¹³C NMR, HRMS-ESI

¹H NMR (400 MHz, DMSO-d₆) δ: 8.02 (d, J = 8.5 Hz, 2H, ArH), 7.86 (d, J = 9.2 Hz, 2H, ArH), 4.60 (s, 2H, CH₂-S), 3.61 (d, J = 2.9 Hz, 2H, NH₂CH₂), 2.09 (d, J = 6.7 Hz, 2H, CH₂-cyclohexyl), 1.79 (s, 3H, Me), 1.66 (m, 7H, Cyclohexyl), 1.54 (m, 4H, cyclohexyl); ¹³C NMR (100 MHz, DMSO-d₆) δ: 190.3, 174.3, 161.5 (2C), 155.9, 142.8, 131.1, 130.0 (2C), 118.7 (2C), 115.3, 43.9, 37.6 (2C), 36.8 (2C), 32.9, 32.6 (3C), 10.9; HRMS-ESI: m/z calcd. for C₂₂H₂₈N₄O₃S [M+H]⁺ 429.194 6, found 429.194 8.

¹H NMR (400 MHz, DMSO-d₆) δ: 8.03 (d, J = 8.5 Hz, 2H, ArH), 7.83 (d, J = 9.9 Hz, 2H, ArH), 4.60 (s, 2H, CH₂-S), 3.60 (d, J = 2.9 Hz, 2H, NH₂CH₂), 2.29-2.26 (m, 2H, CH₂CH₃), 2.08 (d, J = 6.9 Hz, 2H, CH₂-cyclohexyl), 1.65 (m, 7H, cyclohexyl), 1.53 (dd, J = 6.1, 2.1 Hz, 4H, cyclohexyl), 0.91 (s, 3H, CH₂CH₃); ¹³C NMR (100 MHz, DMSO-d₆) δ: 192.3, 174.4, 161.7 (2C), 156.4, 143.2, 131.0, 130.0 (2C), 118.7 (2C), 117.2, 43.9, 37.6, 36.6 (2C), 32.9 (2C), 32.6 (3C), 22.5, 13.7; HRMS-ESI: m/z calcd. for C₂₃H₃₀N₄O₃S [M+H]⁺ 443.210 3, found 443.210 5.

¹H NMR (400 MHz, DMSO-d₆) δ: 8.02 (s, 2H, ArH), 7.88-7.79 (m, 2H, ArH), 4.63-4.55 (m, 2H, CH₂-S), 3.62-3.55 (m, 1H, CH-NH₂), 2.15-2.04 (m, 2H, CH₂-cyclohexyl), 1.83-1.77 (m, 3H, Me), 1.68-1.60 (m, 1H, cyclohexyl), 1.54-1.48 (m, 3H, CHCH₃), 1.35-1.21 (m, 7H, cyclohexyl), 1.02-0.90 (m, 3H, cyclohexyl); ¹³C NMR (100 MHz, DMSO-d₆) δ: 191.9, 174.7, 164.1, 160.9, 156.4, 143.4, 131.0, 129.5 (2C), 118.3 (2C), 114.7, 50.87, 36.6, 36.3, 32.4 (2C), 25.8, 25.5 (3C), 20.7, 10.4; HRMS-ESI: m/z calcd. for C₂₃H₃₀N₄O₃S [M+H]⁺ 443.210 3, found 443.210 6.

¹H NMR (400 MHz, DMSO-d₆) δ: 8.02 (d, J = 8.4 Hz, 2H, ArH), 7.82 (d, J = 8.5 Hz, 2H, ArH), 4.60 (s, 2H, CH₂-S), 3.54 (q, J = 7.1 Hz, 1H, CH-NH₂), 2.28 (q, J = 7.4 Hz, 2H, CH₂CH₃), 2.08 (d, J = 6.7 Hz, 2H, CH₂-cyclohexyl), 1.50-1.40 (m, 3H, CHCH₃), 1.36-1.28 (m, 2H, cyclohexyl), 1.25 (t, J = 6.1 Hz, 5H, cyclohexyl), 0.91 (t, J = 7.2 Hz, 7H, cyclohexyl and CH₂CH₃); ¹³C NMR (100 MHz, DMSO-d₆) δ: 191.9, 174.9, 163.2, 160.44, 156.32, 143.51, 130.96, 129.55 (2C), 118.33 (2C), 114.63, 50.95, 36.76, 36.14, 32.50 (2C), 25.83, 25.55 (3C), 20.87, 17.97, 13.27; HRMS-ESI: m/z calcd. for C₂₄H₃₂N₄O₃S [M+H]⁺ 457.225 9, found 457.226 1.

¹H NMR (400 MHz, DMSO-d₆) δ: 8.07 (s, 2H, ArH), 7.86-7.72 (m, 2H, ArH), 4.60 (s, 2H, CH₂-S), 3.72 (m, 1H, CH-NH₂), 2.15-2.01 (m, 2H, CH₂-cyclohexyl), 1.80-1.74 (s, 3H, Me), 1.71-1.62 (m, 1H, cyclohexyl), 1.52-1.44 (m, 3H, CHCH₃), 1.36-1.09 (m, 7H, cyclohexyl), 1.03-0.90 (m, 3H, cyclohexyl); ¹³C NMR (100 MHz, DMSO-d₆) δ: 191.6, 174.7, 163.2, 160.5, 154.1, 144.0, 131.8, 129.0 (2C), 117.8 (2C), 114.0, 51.0, 36.8, 36.3, 32.8 (2C), 25.9, 25.3 (3C), 18.8, 10.3; HRMS-ESI: m/z calcd. for C₂₃H₃₀N₄O₃S [M+H]⁺ 443.210 3, found 443.210 7.

¹H NMR (400 MHz, DMSO-d₆) δ: 8.04 (d, J = 8.4 Hz, 2H, ArH), 7.80 (d, J = 8.5 Hz, 2H, ArH), 4.61 (s, 2H, CH₂-S), 3.60 (q, J = 7.8 Hz, 1H, CH-NH₂), 2.18 (q, J = 7.2 Hz, 2H, CH₂CH₃), 2.08 (d, J = 7.0 Hz, 2H, CH₂-cyclohexyl), 1.56-1.40 (m, 3H, CHCH₃), 1.35-1.26 (m, 2H, cyclohexyl), 1.21 (t, J = 6.1 Hz, 5H, cyclohexyl), 0.91 (t, J = 7.2 Hz, 7H, cyclohexyl and CH₂CH₃); ¹³C NMR (100 MHz, DMSO-d₆) δ: 191.2, 174.6, 163.8, 161.1, 156.3, 144.7, 131.0, 129.5 (2C), 118.1 (2C), 114.3, 51.8, 36.9 (2C), 35.4 (2C), 31.6, 26.6 (2C), 25.1, 20.2, 18.3, 14.1; HRMS-ESI: m/z calcd. for C₂₄H₃₂N₄O₃S [M+H]⁺ 457.225 9, found 457.226 3.

¹H NMR (400 MHz, DMSO-d₆) δ: 8.09 (d, J = 8.4 Hz, 2H, ArH), 7.82 (d, J = 8.5 Hz, 2H, ArH), 4.68 (s, 2H, CH₂-S), 3.11 (m, 2H, NH₂CH₂CH₂), 2.60 (m, 2H, NH₂CH₂CH₂), 2.28 (m, 2H, CH₂CH₃), 2.08 (d, J = 7.8 Hz, 2H, CH₂-cyclohexyl), 1.56-1.29 (m, 11H, cyclohexyl), 0.91 (t, J = 7.2 Hz, 3H, CH₂CH₃); ¹³C NMR (100 MHz, DMSO-d₆) δ: 191.6, 173.6, 162.1, 160.8, 157.9, 143.6, 131.0, 129.6 (2C), 119.2 (2C), 114.3, 39.8, 36.9, 36.0, 34.5, 33.8 (2C), 32.7, 26.4, 25.3 (2C), 20.2, 14.2; HRMS-ESI: m/z calcd. for C₂₄H₃₂N₄O₃S [M+H]⁺ 457.225 9, found 457.226 0.

¹H NMR (400 MHz, DMSO-d₆) δ: 8.02 (d, J = 8.5 Hz, 2H, ArH), 7.82 (d, J = 8.8 Hz, 2H, ArH), 4.58 (s, 2H, CH₂-S), 3.40 (t, J = 6.6 Hz, 1H, CH-NH₂), 2.07 (s, 3H, Me), 1.79 (m, 4H, CH₂-cyclohexyl and CH₂CH₂CH₂CH₃), 1.66-0.91 (m, 15H, cyclohexyl and CH₂CH₂CH₂CH₃), 0.84 (t, J = 6.9 Hz, 3H, CH₂CH₂CH₂CH₃); ¹³C NMR (100 MHz, DMSO-d₆) δ: 192.0, 174.7, 163.7, 161.0, 156.2, 143.5, 131.0, 129.5 (2C), 118.3 (2C), 114.9, 55.4, 36.6, 36.2, 34.4, 32.4 (2C), 27.3, 25.8 (3C), 25.5, 22.1, 13.9, 10.4; HRMS-ESI: m/z calcd. for C₂₆H₃₆N₄O₃S [M+H]⁺ 485.257 2, found 485.256 8.

¹H NMR (400 MHz, DMSO-d₆) δ: 8.06 (d, J = 8.4 Hz, 2H, ArH), 7.84 (d, J = 8.7 Hz, 2H, ArH), 4.59 (s, 2H, CH₂-S), 3.37 (t, J = 6.5 Hz, 1H, CH-NH₂), 2.28 (q, J = 7.3 Hz, 2H, CH₂CH₃), 2.06 (d, J = 6.7 Hz, 2H, CH₂-cyclohexyl), 1.69-1.17 (m, 13H, cyclohexyl and CH₂CH₂CH₂CH₃), 0.88 (dt, J = 25.0, 7.2 Hz, 10H, CH₂CH₃ and CH₂CH₂CH₂CH₃); ¹³C NMR (100 MHz, DMSO-d₆) δ: 191.9, 174.8, 163.1 (2C), 156.2, 143.5, 130.9, 129.5 (2C), 120.9 (2C), 118.3, 55.5, 36.7, 36.1, 34.5, 32.4 (2C), 27.4, 25.8, 25.5 (2C), 22.1, 17.9, 13.8, 13.2; HRMS-ESI: m/z calcd. for C₂₇H₃₈N₄O₃S [M+H]⁺ 499.272 9, found 499.272 6.

¹H NMR (400 MHz, DMSO-d₆) δ: 8.02 (d, J = 8.4 Hz, 2H, ArH), 7.82 (d, J = 8.8 Hz, 2H, ArH), 4.63 (s, 1H, CH₂-S), 2.78 (t, J = 7.6 Hz, 2H, NH₂CH₂CH₂CH₂CH₂CH₂), 2.40 (t, J = 7.3 Hz, 2H, NH₂CH₂CH₂CH₂CH₂CH₂), 2.28 (d, J = 7.6 Hz, 2H, CH₂-cyclohexyl), 2.08 (s, 3H, Me), 1.66 (q, J = 5.8 Hz, 4H, NH₂CH₂CH₂CH₂CH₂CH₂), 1.61-1.57 (m, 4H, cyclohexyl and NH₂CH₂CH₂CH₂CH₂CH₂), 1.57-1.50 (m, 4H, cyclohexyl), 1.37-1.25 (m, 5H, cyclohexyl); ¹³C NMR (100 MHz, DMSO-d₆) δ: 192.1, 175.5, 164.1 (2C), 155.3, 144.2, 131.0, 129.8 (2C), 118.0 (2C), 117.0, 43, 0, 37.1, 32.9 (2C), 27.2, 26.4 (2C), 25.7 (3C), 24.6, 13.7; HRMS-ESI: m/z calcd. for C₂₆H₃₆N₄O₃S [M+H]⁺ 485.257 2, found 485.257 7.

¹H NMR (400 MHz, DMSO-d₆) δ: 8.00 (d, J = 8.5 Hz, 2H, ArH), 7.82 (d, J = 8.6 Hz, 2H, ArH), 4.60 (s, 2H, CH₂-S), 2.76 (t, J = 7.6 Hz, 2H, NH₂CH₂CH₂CH₂CH₂CH₂), 2.40 (t, J = 7.3 Hz, 2H, NH₂CH₂CH₂CH₂CH₂CH₂), 2.30 (m, 2H, CH₂CH₃), 2.08 (d, J = 6.8 Hz, 2H, CH₂-cyclohexyl), 1.66 (q, J = 5.8 Hz, 4H, NH₂CH₂CH₂CH₂CH₂CH₂), 1.61-1.57 (m, 4H, cyclohexyl), 1.57-1.50 (m, 4H, cyclohexyl and NH₂CH₂CH₂CH₂CH₂CH₂), 1.37-1.25 (m, 5H, cyclohexyl), 0.92 (m, 3H, CH₂CH₃); ¹³C NMR (100 MHz, DMSO-d₆) δ: 192.3, 175.8, 164.3 (2C), 155.2, 144.5, 131.1, 129.9 (2C), 118.6 (2C), 117.2, 42.8, 37.2 (2C), 32.9 (2C), 27.2, 26.2 (2C), 25.9 (2C), 24.9 (2C), 18.4, 13.7; HRMS-ESI: m/z calcd. for C₂₇H₃₈N₄O₃S [M+H]⁺ 499.272 9, found 499.273 0.

¹H NMR (400 MHz, DMSO-d₆) δ: 8.03 (d, J = 8.4 Hz, 2H, ArH), 7.84 (d, J = 8.6 Hz, 2H, ArH), 4.60 (s, 2H, CH₂-S), 3.21 (d, J = 5.5 Hz, 1H, CH-NH₂), 2.28 (q, J = 7.4 Hz, 2H, CH₂CH₃), 2.07 (d, J = 6.8 Hz, 2H, CH₂-cyclohexyl), 1.94 (dq, J = 13.4, 6.8 Hz, 1H, CH(CH₃)₂), 1.49-1.39 (m, 6H, CH(CH₃)₂), 1.30-1.20 (m, 3H, CH₂CH₃), 0.89 (m, 11H, cyclohexyl); ¹³C NMR (100 MHz, DMSO-d₆) δ: 191.8, 174.3, 163.0, 160.8, 156.1, 143.3, 130.8, 129.4 (2C), 120.8 (2C), 118.2, 60.6, 36.6, 35.9, 32.3 (2C), 31.7, 30.1, 25.7, 25.4 (2C), 19.3, 17.8, 17.2, 13.2; HRMS-ESI: m/z calcd. for C₂₆H₃₆N₄O₃S [M+H]⁺ 485.257 2, found 485.257 5.

¹H NMR (400 MHz, DMSO-d₆) δ: 8.03 (d, J = 8.4 Hz, 2H, ArH), 7.83 (d, J = 8.5 Hz, 2H, ArH), 4.59 (s, 2H, CH₂-S), 3.25 (d, J = 5.6 Hz, 1H, CH-NH₂), 2.07 (d, J = 6.7 Hz, 2H, CH₂-cyclohexyl), 1.96 (m, 1H, CH(CH₃)₂), 1.80 (s, 3H, Me), 1.61 (d, J = 21.3 Hz, 1H, cyclohexyl), 1.45 (d, J = 10.2 Hz, 4H, cyclohexyl), 1.32-1.19 (m, 4H, cyclohexyl), 1.05 (t, J = 7.0 Hz, 2H, cyclohexyl), 0.93 (d, J = 6.7 Hz, 3H, CH(CH₃)₂), 0.86 (d, J = 6.7 Hz, 3H, CH(CH₃)₂); ¹³C NMR (100 MHz, DMSO-d₆) δ: 192.0, 174.0, 163.6, 161.0, 156.0, 143.4, 131.0, 129.5 (2C), 120.5 (2C), 118.3, 60.5, 36.6, 36.2 (2C), 32.4 (2C), 31.7, 25.8, 25.5 (2C), 18.5 (2C), 10.42; HRMS-ESI: m/z calcd. for C₂₅H₃₄N₄O₃S [M+H]⁺ 471.241 6, found 471.242 0.

¹H NMR (400 MHz, DMSO-d₆) δ: 8.03 (d, J = 8.9 Hz, 2H, ArH), 7.89-7.79 (m, 2H, ArH), 4.58 (s, 2H, CH₂-S), 3.27-3.20 (m, 1H, CH-NH₂), 2.06 (q, J = 5.7 Hz, 2H, CH₂-cyclohexyl), 1.95 (m, 1H, CH(CH₃)₂), 1.83-1.76 (m, 3H, Me), 1.52-1.18 (m, 6H, CH(CH₃)₂), 1.04-0.79 (m, 11H, cyclohexyl); ¹³C NMR (100 MHz, DMSO-d₆) δ: 192.0, 174.1, 163.6 (2C), 156.1, 143.4, 130.9, 129.5 (2C), 118.3 (2C), 114.8, 60.6, 36.6, 36.2, 32.4 (2C), 31.7 (2C), 25.8, 25.5 (2C), 19.4 (2C), 10.4; HRMS-ESI: m/z calcd. for C₂₅H₃₄N₄O₃S [M+H]⁺ 471.241 6, found 471.241 8.

¹H NMR (400 MHz, DMSO-d₆) δ: 8.02 (d, J = 8.4 Hz, 2H, ArH), 7.83 (d, J = 8.8 Hz, 2H, ArH), 4.58 (s, 2H, CH₂-S), 3.19 (d, J = 5.5 Hz, 1H, CH-NH₂), 2.28 (q, J = 7.3 Hz, 2H, CH₂CH₃), 2.07 (d, J = 6.8 Hz, 2H, CH₂-cyclohexyl), 1.98-1.88 (m, 1H, CH(CH₃)₂), 1.65-1.19 (m, 9H, CH(CH₃)₂ and CH₂CH₃), 0.88 (m, 11H, cyclohexyl); ¹³C NMR (100 MHz, DMSO-d₆) δ: 192.0, 174.5, 163.6 (2C), 156.1, 143.5, 130.9, 129.5 (2C), 118.3 (2C), 114.6, 60.8, 36.6, 36.1, 32.5 (2C), 31.8 (2C), 25.8, 25.5 (2C), 19.4, 17.9, 17.2, 13.3; HRMS-ESI: m/z calcd. for C₂₆H₃₆N₄O₃S [M+H]⁺ 485.257 2, found 485.257 0.

¹H NMR (400 MHz, DMSO-d₆) δ: 8.01 (d, J = 8.7 Hz, 2H, ArH), 7.83 (d, J = 8.2 Hz, 2H, ArH), 4.57 (s, 2H, CH₂-S), 3.43-3.40 (m, 1H, CH-NH₂), 2.08 (d, J = 6.7 Hz, 2H, CH₂-cyclohexyl), 1.77-1.70 (m, 1H, CH₂CH(CH₃)₂), 1.61 (m, 8H, Me and cyclohexyl), 1.54-1.51 (m, 5H, cyclohexyl and CH₂CH(CH₃)₂), 1.17 (m, 3H, cyclohexyl), 0.91 (m, 6H, CH₂CH(CH₃)₂); ¹³C NMR (100 MHz, DMSO-d₆) δ: 192.2. 175.8. 160.9. 144.0, 131.4, 131.2, 129.9 (2C), 118.8 (2C), 112.9, 54.5, 44.4, 36.7 (2C), 33.1 (2C), 33.0, 26.3, 26.1 (2C), 23.2, 22.6 (2C), 13.8; HRMS-ESI: m/z calcd. for C₂₆H₃₆N₄O₃S [M+H]⁺ 485.257 2, found 485.257 5.

¹H NMR (400 MHz, DMSO-d₆) δ: 8.06 (d, J = 8.7 Hz, 2H, ArH), 7.85 (t, J = 8.2 Hz, 2H, ArH), 4.59 (s, 2H, CH₂-S), 3.44-3.41 (m, 1H, CH-NH₂), 2.29 (m, 2H, CH₂CH₃), 2.08 (d, J = 6.7 Hz, 2H, CH₂-cyclohexyl), 1.77-1.70 (m, 1H, CH₂CH(CH₃)₂), 1.64-1.60 (m, 8H, cyclohexyl), 1.54-1.50 (m, 5H, cyclohexyl and CH₂CH(CH₃)₂), 1.17 (t, J = 7.1 Hz, 3H, CH₂CH₃), 0.92 (m, 6H, CH₂CH(CH₃)₂); ¹³C NMR (100 MHz, DMSO-d₆) δ: 192.2, 175.8, 160.9, 144.0, 131.4, 131.2 (2C), 129.9 (2C), 118.8, 112.9, 54.5, 44.4, 36.7 (2C), 33.1 (2C), 33.0, 26.3, 26.1 (2C), 23.2, 22.6 (2C), 18.5, 13.8; HRMS-ESI: m/z calc.d for C₂₇H₃₈N₄O₃S [M+H]⁺ 499.272 9, found 499.273 3.

¹H NMR (400 MHz, DMSO-d₆) δ: 8.02 (d, J = 8.4 Hz, 2H, ArH), 7.83 (d, J = 8.7 Hz, 2H, ArH), 4.59 (s, 2H, CH₂-S), 3.44 (dd, J = 8.5, 5.8 Hz, 1H, CH-NH₂), 2.08 (d, J = 6.6 Hz, 2H, CH₂-cyclohexyl), 1.80 (s, 3H, Me), 1.77-1.69 (m, 1H, CH₂CH(CH₃)₂), 1.54-1.20 (m, 10H, cyclohexyl and CH₂CH(CH₃)₂), 0.89 (m, 9H, cyclohexyl and CH₂CH(CH₃)₂); ¹³C NMR (100 MHz, DMSO-d₆) δ: 192.5, 175.5, 163.3, 161.4, 156.1, 144.0, 130.9, 129.9 (2C), 120.5 (2C), 118.4, 54.2, 44.2, 37.1, 36.7, 32.8 (2C), 30.1, 25.9 (2C), 25.8, 23.5, 22.4 (2C), 13.8; HRMS-ESI: m/z calcd. for C₂₆H₃₆N₄O₃S [M+H]⁺ 485.257 2, found 485.257 6.

¹H NMR (400 MHz, DMSO-d₆) δ: 8.06-7.99 (m, 2H, ArH), 7.82 (t, J = 6.2 Hz, 2H, ArH), 4.60 (d, J = 4.0 Hz, 2H, CH₂-S), 3.43-3.39 (m, 1H, CH-NH₂), 2.28 (m, 2H, CH₂CH₃), 2.08 (d, J = 5.5 Hz, 2H, CH₂-cyclohexyl), 1.73 (dp, J = 13.4, 6.6 Hz, 1H, CH₂CH(CH₃)₂), 1.51-1.23 (m, 9H, cyclohexyl and CH₂CH(CH₃)₂), 0.97-0.83 (m, 13H, cyclohexyl and CH₂CH(CH₃)₂) and CH₂CH₃); ¹³C NMR (100 MHz, DMSO-d₆) δ: 191.9, 175.3, 163.2, 160.5, 156.3, 143.6, 130.9, 129.6 (2C), 120.9 (2C), 118.4, 54.1, 43.9, 36.7, 36.1, 32.5 (2C), 25.8, 25.6 (2C), 24.2, 23.1 (2C), 21.9, 17.9, 13.3; HRMS-ESI: m/z calcd. for C₂₇H₃₈N₄O₃S [M+H]⁺ 499.272 9, found 499.272 7.

¹H NMR (400 MHz, DMSO-d₆) δ: 8.02 (d, J = 8.4 Hz, 2H, ArH), 7.80 (d, J = 8.6 Hz, 2H, ArH), 7.27-7.16 (m, 5H, ArH), 4.61 (s, 2H, CH₂-S), 3.66 (m, 1H, CH-NH₂), 3.01 (dd, J = 13.4, 5.7 Hz, 1H, CH₂-Ph), 2.77 (dd, J = 13.4, 7.7 Hz, 1H, CH₂-Ph), 2.09 (d, J = 6.6 Hz, 2H, CH₂-cyclohexyl), 1.80 (s, 3H, Me), 1.26 (t, J = 14.8 Hz, 7H, cyclohexyl), 0.92 (h, J = 13.4, 12.3 Hz, 4H, cyclohexyl); ¹³C NMR (100 MHz, DMSO-d₆) δ: 192.3, 174.4, 163.9 (2C), 156.4, 143.9, 138.7, 131.4, 130.0, 129.7 (2C), 128.5 (2C), 126.6 (2C), 118.8 (2C), 115.4, 57.5, 37.1, 36.7 (2C), 32.9 (2C), 26.3, 26.0 (3C), 10.8; HRMS-ESI: m/z calcd. for C₂₉H₃₄N₄O₃S [M+H]⁺ 519.241 6, found 519.242 0.

¹H NMR (400 MHz, DMSO-d₆) δ: 8.02 (d, J = 8.4 Hz, 2H, ArH), 7.81 (d, J = 8.4 Hz, 2H, ArH), 7.26-7.16 (m, 5H, ArH), 4.61 (s, 2H, CH₂-S), 3.69 (m, 1H, CH-NH₂), 3.02 (dd, J = 13.4, 5.7 Hz, 1H, CH₂-Ph), 2.77 (dd, J = 13.4, 7.7 Hz, 1H, CH₂-Ph), 2.29 (q, J = 7.3 Hz, 2H, CH₂CH₃), 2.08 (d, J = 6.8 Hz, 2H, CH₂-cyclohexyl), 1.52-1.18 (m, 7H, cyclohexyl), 0.91 (t, J = 7.6 Hz, 7H, cyclohexyl and CH₂CH₃); ¹³C NMR (100 MHz, DMSO-d₆) δ: 192.3, 174.4, 163.6, 160.9, 156.7, 143.9, 138.7, 131.4, 129.9 (2C), 129.8 (2C), 128.5 (2C), 126.6, 121.4 (2C), 118.9, 57.5, 37.2, 36.6 (2C), 32.9 (2C), 26.3, 26.0 (3C), 18.4, 13.7; HRMS-ESI: m/z calcd. for C₃₀H₃₆N₄O₃S [M+H]⁺ 533.257 2, found 533.257 0.

¹H NMR (400 MHz, DMSO-d₆) δ: 10.44 (s, 1H, C=O-NH), 8.02 (d, J = 8.4 Hz, 2H, ArH), 7.85 (d, J = 8.5 Hz, 2H, ArH), 4.59 (s, 2H, CH₂-S), 3.80 (m, 1H, pyrrolidinyl), 2.92 (t, J = 6.6 Hz, 2H, pyrrolidinyl), 2.08 (d, J = 7.0 Hz, 2H, CH₂-cyclohexyl), 1.79 (s, 3H, Me), 1.73-1.18 (m, 11H, cyclohexyl), 0.91 (m, 4H, pyrrolidinyl); ¹³C NMR (100 MHz, DMSO-d₆) δ: 192.5, 174.0, 164.1, 161.4, 156.5, 143.5, 131.6, 130.0 (2C), 118.8 (2C), 115.4, 61.2, 47.1, 37.1, 36.7, 32.8 (2C), 30.9 (2C), 26.3, 26.1 (2C), 25.9, 10.9; HRMS-ESI: m/z calcd. for C₂₅H₃₂N₄O₃S [M+H]⁺ 469.225 9, found 469.226 3.

¹H NMR (400 MHz, DMSO-d₆) δ: 10.69 (s, 1H, C=O-NH), 8.20-7.70 (m, 4H, ArH), 4.59 (s, 2H, CH₂-S), 3.90 (s, 1H, pyrrolidinyl), 2.95 (m, 2H, pyrrolidinyl), 2.16 (m, 4H, CH₂-cyclohexyl and CH₂CH₃), 1.72-1.10 (m, 12H, cyclohexyl and pyrrolidinyl), 1.01-0.75 (m, 6H, pyrrolidinyl and CH₂CH₃); ¹³C NMR (100 MHz, DMSO-d₆) δ: 192.5, 173.2, 163.8, 160.8, 156.8, 143.5, 131.6, 129.9 (2C), 121.3 (2C), 118.8, 61.0, 46.9, 43.9, 37.2, 36.5, 32.9, 30.8, 26.3, 25.9, 25.8, 22.5, 22.2, 18.4, 13.7; HRMS-ESI: m/z calcd. for C₂₆H₃₄N₄O₃S [M+H]⁺ 483.241 6, found 483.241 8.

¹H NMR (400 MHz, DMSO-d₆) δ: 10.69 (s, 1H, C=O-NH), 8.20 (d, J = 8.4 Hz, 2H, ArH), 7.80 (d, J = 8.6 Hz, 2H, ArH), 4.60 (s, 2H, CH₂-S), 3.90 (t, 1H, pyrrolidinyl), 2.95 (m, 2H, pyrrolidinyl), 2.16 (q, J = 8.5 Hz, 2H, CH₂CH₃), 2.06(d, J = 7.2, 2H, CH₂-cyclohexyl), 1.72-1.10 (m, 12H, cyclohexyl and pyrrolidinyl), 1.01-0.75 (m, 6H, pyrrolidinyl and CH₂CH₃); ¹³C NMR (100 MHz, DMSO-d₆) δ: 191.4, 172.2, 163.8, 159.8, 156.8, 146.2, 131.8, 129.9 (2C), 121.3 (2C), 118.8, 62.4, 46.9, 43.9, 37.2, 36.5, 32.9, 30.8, 26.3, 26.0, 25.8, 22.6, 22.3, 18.4, 14.0; HRMS-ESI: m/z calcd. for C₂₆H₃₄N₄O₃S [M+H]⁺ 483.241 6, found 483.242 0.

Compd.	R¹	CC₅₀ /µmol·L^-1	EC₅₀ /µmol·L^-1	TI
2a	Me	> 200.00	2.59 ± 0.95	> 77.31
2b	Et	> 200.00	0.15 ± 0.02	> 1 375.09
2c	Me	17.32 ± 2.60	0.09 ± 0.01	197.78
2d	Et	47.52 ± 0.25	0.02 ± 0.01	2 159.67
2e	Me	150.60 ± 6.09	1.54 ± 0.05	97.76
2f	Et	> 200.00	0.47 ± 0.12	> 429.78
2g	Et	106.68 ± 17.93	0.87 ± 0.18	122.09
2h	Me	41.17 ± 2.03	0.13 ± 0.05	316.79
2i	Et	50.53 ± 8.03	0.02 ± 0.01	2 797.55
2j	Me	69.70 ± 5.40	4.04 ± 0.79	17.27
2k	Et	89.31 ± 4.39	0.13 ± 0.03	684.40
2l	Et	22.61 ± 3.11	0.03 ± 0.00	916.56
2m	Me	31.75 ± 0.46	0.55 ± 0.02	57.99
2n	Me	15.70 ± 1.41	0.08 ± 0.00	187.32
2o	Et	62.09 ± 10.20	0.06 ± 0.01	1 014.93
2p	Me	74.80 ± 9.31	0.66 ± 0.18	112.94
2q	Et	66.34 ± 4.12	0.12 ± 0.01	550.22
2r	Me	45.15 ± 4.28	0.11 ± 0.06	401.64
2s	Et	55.80 ± 1.46	0.02 ± 0.01	3 401.77
2t	Me	18.77 ± 0.42	0.23 ± 0.02	80.04
2u	Et	35.03 ± 10.07	0.05 ± 0.01	663.13
2v	Me	24.74 ± 1.18	0.27 ± 0.02	91.20
2w	Et	59.62 ± 0.79	0.01 ± 0.01	4 881.85
2x	Et	44.30 ± 8.79	0.13 ± 0.07	333.42
DB02		> 200.00	0.03 ± 0.01	> 7 750.81
3TC		> 200.00	0.61 ± 0.72	> 325.42

Compd.

R¹

R²

CC₅₀ /µmol·L^-1

EC₅₀ /µmol·L^-1

> 200.00

2.59 ± 0.95

> 77.31

> 200.00

0.15 ± 0.02

> 1 375.09

17.32 ± 2.60

0.09 ± 0.01

197.78

47.52 ± 0.25

0.02 ± 0.01

2 159.67

150.60 ± 6.09

1.54 ± 0.05

97.76

> 200.00

0.47 ± 0.12

> 429.78

106.68 ± 17.93

0.87 ± 0.18

122.09

41.17 ± 2.03

0.13 ± 0.05

316.79

50.53 ± 8.03

0.02 ± 0.01

2 797.55

69.70 ± 5.40

4.04 ± 0.79

17.27

89.31 ± 4.39

0.13 ± 0.03

684.40

22.61 ± 3.11

0.03 ± 0.00

916.56

31.75 ± 0.46

0.55 ± 0.02

57.99

15.70 ± 1.41

0.08 ± 0.00

187.32

62.09 ± 10.20

0.06 ± 0.01

1 014.93

74.80 ± 9.31

0.66 ± 0.18

112.94

66.34 ± 4.12

0.12 ± 0.01

550.22

45.15 ± 4.28

0.11 ± 0.06

401.64

55.80 ± 1.46

0.02 ± 0.01

3 401.77

18.77 ± 0.42

0.23 ± 0.02

80.04

35.03 ± 10.07

0.05 ± 0.01

663.13

24.74 ± 1.18

0.27 ± 0.02

91.20

59.62 ± 0.79

0.01 ± 0.01

4 881.85

44.30 ± 8.79

0.13 ± 0.07

333.42

DB02

> 200.00

0.03 ± 0.01

> 7 750.81

3TC

> 200.00

0.61 ± 0.72

> 325.42