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L. Tomaškovic et al. / Bioorg. Med. Chem. 21 (2013) 321–332
328
0.59 mmol), amine 33 (230 mg; 0.29 mmol) and EDCꢁHCl
(235 mg; 1.23 mmol) were added. The reaction mixture was stirred
at room temperature overnight (18 h). The solvent was evaporated
under reduced pressure and the residue purified on a silica gel col-
umn (eluant CHCl3/MeOH/NH4OH, 6:1:0.1). 224 mg (67%) of com-
pound 44 were obtained. MS (ESI) 1152.49 (MH+), 577.07
[(MH2)++]. HRMS (ESI) calcd for C61H104FN3O16 [(MH2)++]:
576.8700; found 576.8698. IR (KBr) 3517 and 3433, 3600–3100
(O–H, N–H), 2975, 2943 and 2871 (C–H), 1722 (C@O lactone),
1665 (C@O ketone), 1647 (C@O amide), 1624 and 1606 (C@C),
1560, 1535, 1459, 1439, 1420, 1377, 1321, 1268, 1223, 1176,
1166, 1127, 1108, 1094, 1054, 1004, 956, 895, 796, 752, 701, 670
(cmꢀ1). 1H NMR (500 MHz, DMSO-d6) d 7.34 (NH), 7.31 (H-1S),
6.21 (H-2S), 5.99 (H-4S), 4.83 (H-13), 4.79 (H-100), 4.42 (H-10), 4.11
(H-11S), 4.07 (H-500), 4.01 (H-3), 3.66 (H-50), 3.51 (H-5, H-11),
3.22 (300ꢀOCH3), 3.07 (HaꢀCONHCH2CH2CH2), 3.04 (H-20, H-16S),
3.00 (Hb-CONHCH2CH2CH2), 2.91 (H-400), 2.82 (Ha–CON-
HCH2CH2CH2), 2.71 (H-2, H-10), 2.62 (Ha-6S), 2.55 (Ha-9), 2.47
(H-30, Hb–CONHCH2CH2CH2), 2.27 (Ha-200), 2.34 (H-8S), 2.31 (Hb-
6S), 2.22 [30–N(CH3)2], 2.07 (Hb-9), 2.03 (H-14S), 2.03 (Ha-12S),
1.94 (H-8), 1.91 (H-4), 1.77 (Ha-13–CH2, Ha-7S), 1.67 (Ha–CON-
HCH2CH2CH2), 1.61 (Ha-7), 1.59 (Ha-40, Ha-15S), 1.57 (Hb–CON-
HCH2CH2CH2), 1.50 (Hb-200), 1.48 (10S–CH3), 1.42 (Hb-12S), 1.35
(Hb-15S), 1.34 (Hb-13–CH2, Hb-7S), 1.20 (6–CH3), 1.15 (500–CH3),
1.13 (300–CH3), 1.09 (10–CH3), 1.07 (50–CH3), 1.04 (Hb-7, Hb-40),
1.02 (12–CH3), 1.01 (2–CH3), 0.98 (4–CH3), 0.95 (13S–CH3), 0.84
(8–CH3), 0.80 (16S–CH3), 0.79 (13–CH3). 13C NMR (125 MHz,
5.1.6. Preparation of compound 46
Compound 46 (107 mg, 32%) was prepared from acid 14
(110 mg; 0.29 mmol) and amine 33 (230 mg; 0.29 mmol). MS
(ESI) 1153.5 (MH+), 577.3 [(MH2)++]. HRMS (ESI) Calcd for
C
61H104FN3O16 [(MH2)++]: 576.8700; found 576.8690. IR (KBr)
3434, 3600–3100 (O–H, N–H), 2973, 2937 and 2875 (C–H), 1727
(C@O lactone), 1666 (C@O ketone), 1629 and 1609 (C@C), 1520,
1456, 1378, 1318, 1281, 1271, 1167, 1110, 1094, 1054, 1014,
958, 899, 823, 716 (cmꢀ1). 1H NMR (500 MHz, CDCl3) d 7.28 (H-
1S), 6.29 (H-4S), 6.27 (H-2S), 5.31 (H-6S), 4.97 (H-100), 4.77 (H-13),
4.49 (H-10), 4.46 (H-11S), 4.10 (H-3), 3.58 (H-5), 3.32 (300–OCH3),
2.48 [30–N(CH3)2], 1.43 (10S–CH3), 1.36 (6–CH3), 1.10 (13S–CH3),
0.92 (16S–CH3), 0.89 (13–CH3). 13C NMR (75 MHz, CDCl3)
185.74, 178.40, 155.66, 127.86, 118.02, 102.83, 96.45, 87.71,
83.74, 79.31, 77.74, 69.50, 49.43, 40.52, 26.87, 21.16, 17.49,
14.68, 11.11.
d
5.1.7. Preparation of compound 47
Compound 47 (170 mg, 61%) was prepared from acid 17
(96 mg; 0.24 mmol) and amine 33 (197 mg; 0.24 mmol). MS (ESI)
1169.2 (MH+), 585.3 [(MH2)++]. HRMS (ESI) Calcd for
C
61H104ClN3O16 [(MH2)++]: 584.8553; found 584.8555. IR (KBr)
3422, 3600-3100 (O–H, N–H), 2971, 2938 and 2874 (C–H), 1732
(C@O lactone), 1662 (C@O ketone), 1619 and 1609 (C@C), 1512,
1454, 1378, 1317, 1247, 1168, 1111, 1052, 1013, 959, 886, 836,
755 (cmꢀ1). 1H NMR (500 MHz, CDCl3) d 7.29 (H-1S), 6.29 (H-2S),
6.07 (H-4S), 4.98 (H-100), 4.72 (H-13), 4.54 (H-11S), 4.46 (H-10),
4.10 (H-3), 3.58 (H-5), 3.32 (300–OCH3), 2.38 [30–N(CH3)2], 1.67
(10S–CH3), 1.36 (6–CH3), 1.10 (13S–CH3), 1.08 (16S–CH3), 0.88
(13–CH3). 13C NMR (75 MHz, CDCl3) d 186.76, 177.83, 172.89,
152.89, 129.25, 124.93, 102.97, 95.97, 82.61, 79.26, 76.38, 75.51,
49.48, 40.51, 26.70, 24.41, 20.58, 18.38, 11.12.
DMSO-d6)
d 186.19, 177.32, 173.13, 168.11, 153.85, 129.79,
124.98, 102.96, 102.41, 95.83, 87.24, 83.49, 78.79, 78.35, 77.39,
75.99, 75.24, 74.42, 73.66, 71.93, 71.54, 68.03, 65.80, 65.61,
63.71, 60.74, 49.71, 48.96, 48.40, 45.14, 43.94, 41.48, 41.23,
38.45, 36.40, 35.74, 35.74, 35.73, 34.78, 32.90, 31.25, 30.96,
28.21, 28.19, 28.05, 28.05, 23.79, 23.30, 22.28, 22.35, 21.91,
19.37, 18.19, 17.78, 16.00, 15.76, 11.91, 10.44, 10.29.
5.1.8. Preparation of compound 48
Compound 48 (258 mg, 76%) was prepared from acid 15
(115 mg; 0.29 mmol) and amine 33 (230 mg; 0.29 mmol). MS
(ESI) 1171.0 (MH+), 586.1 [(MH2)++]. HRMS (ESI) Calcd for
5.1.5. Preparation of compound 45
Compound 45 (224 mg, 89%) was prepared from acid 13
(80 mg; 0.22 mmol) and amine 33 (174 mg; 0.22 mmol). MS (ESI)
1136.53 (MH+), 569.08 [(MH2)++]. HRMS (ESI) Calcd for
C
61H103F2N3O16 [(MH2)++]: 585.8653; found 585.8641. IR (KBr)
3433, 3600–3100 (O–H, N–H), 2969, 2938 and 2876 (C–H), 1728
(C@O lactone), 1671 (C@O ketone), 1634, 1524, 1456, 1378,
1316, 1283, 1167, 1110, 1055, 1032, 999, 960, 933, 899, 821,
728, 711 (cmꢀ1). 1H NMR (500 MHz, CDCl3) d 7.20 (H-1S), 6.40
(H-4S), 6.33 (H-2S), 5.38 (H-6S), 5.03 (H-100), 4.67 (H-13), 4.45(H-
10), 4.33 (H-11S), 4.09 (H-3), 3.60 (H-5), 3.33 (300–OCH3), 2.30 [30–
N(CH3)2], 1.53 (10S–CH3), 1.35 (6–CH3), 1.10 (13S–CH3), 0.94
(16S–CH3), 0.88 (13–CH3). 13C NMR (75 MHz, CDCl3) d 185.86,
178.70, 172.77, 151.50, 129.85, 120.97, 102.87, 95.72, 86.82,
83.01, 79.39, 77.84, 71.76, 49.50, 40.34, 27.15, 23.04, 16.52,
14.65, 11.08.
C
61H104FN3O15 [(MH2)++]: 568.8726; found 568.8716. IR (KBr)
3513 and 3423, 3600–3100 (O–H, N–H), 2972, 2942 and 2873
(C–H), 1720 (C@O lactone), 1666 (C@O ketone), 1647 (C@O amide),
1630 and 1610 (C@C), 1561, 1534, 1459, 1377, 1294, 1271, 1248,
1166, 1109, 1094, 1054, 1005, 958, 890, 829, 752 (cmꢀ1). 1H
NMR (500 MHz, DMSO-d6) d 7.44 (NH), 7.30 (H-1S), 6.21 (H-2S),
6.00 (H-4S), 4.89 (H-13), 4.79 (H-100), 4.42 (H-10), 4.08 (H-11S),
4.08 (H-500), 4.01 (H-3), 3.66 (H-50), 3.51 (H-5, H-11), 3.22 (300–
OCH3), 3.02 (H–CONHCH2CH2CH2, H-20), 2.91 (H-400), 2.82 (Ha–
CONHCH2CH2CH2), 2.71 (H-2, H-10), 2.63 (Ha-6S), 2.53 (Ha-9),
2.50 (H-16S), 2.44 (H-30, Hb–CONHCH2CH2CH2), 2.32 (Hb-6S), 2.29
(H-8S), 2.27 (Ha-200), 2.23 [30–N(CH3)2], 2.09 (Hb-9), 1.95 (H-4),
1.90 (H-8), 1.77 (Ha-13–CH2, Ha-7S), 1.77 (Ha-12S), 1.64 (Ha–CON-
HCH2CH2CH2, Ha-7), 1.63 (Hb-12S), 1.60 (Ha-15S), 1.59 (Ha-40),
1.51 (Hb–CONHCH2CH2CH2, Hb-200), 1.49 (10S–CH3), 1.37 (Hb-13–
CH2), 1.35 (Hb-7S, Hb-15S), 1.20 (6–CH3), 1.15 (Hb-7, 500–CH3),
1.14 (300–CH3), 1.10 (2–CH3), 1.07 (50–CH3), 1.04 (Hb-40), 1.01 (12–
CH3), 1.00 (10–CH3), 0.98 (4–CH3), 0.90 (13S–CH3), 0.88 (8–CH3,
16S–CH3), 0.80 (13–CH3). 13C NMR (75 MHz, DMSO-d6) d 185.15,
176.32, 170.75, 166.96, 152.80, 128.83, 124.04, 101.89, 101.44,
94.80, 82.45, 77.77, 77.27, 76.24, 74.93, 74.42, 74.24, 72.65,
70.54, 70.14, 66.97, 64.78, 64.76, 64.55, 62.84, 59.88, 48.71,
48.02, 48.37, 44.17, 43.86, 42.57, 40.40, 40.26, 37.20, 34.69,
33.32, 32.60, 30.90, 30.13, 29.96, 27.91, 27.07, 27.07, 27.07,
27.07, 22.34, 22.33, 21.33, 21.20, 20.88, 18.36, 18.35, 18.11,
15.72, 14.83, 10.92, 9.31, 9.26.
5.1.9. Preparation of compound 49
Compound 49 (105 mg, 69%) was prepared from acid 25
(50 mg; 0.13 mmol) and amine 33 (115 mg; 0.14 mmol). MS (ESI)
1155.2 (MH+), 578.3 [(MH2)++]. HRMS (ESI) Calcd for
C
61H103F2N3O15 [(MH2)++]: 577.8679; found 577.8678. IR (KBr)
3444, 3600–3100 (O–H, N–H), 2969, 2940, 2876 and 2782 (C–H),
1731 (C@O lactone), 1671 (C@O ketone), 1634, 1525, 1456, 1379,
1317, 1293, 1263, 1176, 1167, 1110, 1057, 1015, 959, 938, 899,
835, 816, 756, 709 (cmꢀ1). 1H NMR (500 MHz, CDCl3) d 7.29 (H-
1S), 6.39 (H-4S), 6.34 (H-2S), 5.37 (H-6S), 4.98 (H-100), 4.79 (H-13),
4.48 (H-10), 4.30 (H-11S), 4.10 (H-3), 3.61 (H-5), 3.32 (300–OCH3),
2.42 [30–N(CH3)2], 1.53 (10S–CH3), 1.39 (6–CH3), 1.02 (13S–CH3),
0.98 (16S–CH3), 0.90 (13–CH3). 13C NMR (75 MHz, CDCl3)
185.31, 178.00, 171.63, 151.13, 129.25, 120.31, 102.44, 95.30,
d