4216
B. Choucair et al. / Bioorg. Med. Chem. Lett. 14 (2004) 4213–4216
6. Moriarty, R. M.; Tuladhar, S. M.; Guo, L.; Wehrli, S.
Tetrahedron Lett. 1994, 35, 8103.
3H, Me 18); 0.86 (d, 6H, J ¼ 6:5, Me 27 and Me 26); 0.90
(d, 3H, J ¼ 6:5, Me 21); 0.93 (s, 3H, Me 19); 2.37 (m, 1H,
0
St–NH–CHaHa –(CH2)2–); 2.42–2.47 (m, 2H, –HN–
7. Zhang, X.; Rao, M. N.; Jones, S. R.; Shao, B.; Feibush,
P.; McGuigan, M.; Tzodikov, N.; Feibush, B.; Sharkan-
sky, I.; Snyder, B.; Mallis, L. M.; Sarkahian, A.; Wilder,
S.; Turse, J. E.; Kinney, W. A. J. Org. Chem. 1998, 63,
8599.
8. Jones, S. R.; Selinsky, B. S.; Rao, M. N.; Zhang, X.;
Kinney, W. A.; Tham, F. S. J. Org. Chem. 1998, 63, 3786.
9. Pechulis, A. D.; Bellevue, F. H.; Cioffi, C. L.; Trapp, S. G.;
Fojtik, J. P.; McKitty, A. A.; Kinney, W. A.; Frye, L. L.
J. Org. Chem. 1995, 60, 5121.
10. Zhou, X.-D.; Cai, F.; Zhou, W.-S. Tetrahedron 2002, 58,
10293.
11. Moriarty, R. M.; Enache, L. A.; Kinney, W. A.; Allen, C.
S.; Canary, J. W.; Tuladhar, S. M.; Guo, L. Tetrahedron
Lett. 1995, 36, 5139.
12. Sadownik, A.; Deng, G.; Janout, V.; Regen, S. L.;
Bernard, E. M.; Kikuchi, K.; Armonstrong, D. J. Am.
Chem. Soc. 1995, 117, 6138.
13. Jones, S. R.; Kinney, W. A.; Zhang, X.; Jones, L. M.;
Selinsky, B. S. Steroids 1996, 61, 565.
14. Shu, Y.; Jones, S. R.; Kinney, W. A.; Selinsky, B. S.
Steroids 2002, 67, 291.
15. El Kihel, L.; Soustre, I.; Karst, F.; Letourneux, Y. FEMS
Microbiol. Lett. 1994, 120, 163.
(CH2)3–CH2–NH2 et –HN-CH2–(CH2)3–NH2); 2.70 (m,
0
1H, St–NH–CHaHa –(CH2)2–); 2.65 (dd, 1H, J7a–8 ¼ 6:8,
J7a–6 ¼ 1:3, H7a of b epimer); 2.79 (t, 2H, J ¼ 8:3, St–
NH–(CH2)2–CH2–NH–); 3.5 (m, 1H, H3a); 5.30 (d, 1H,
J ¼ 3:0, H6 of b epimer); 7.19 (br s, 2H, –NH2, D2O
exchange); 8.13 (br s, 1H, –NH–, D2O exchange); 8.39 (br
s, 1H, –NH–, D2O exchange). 13C NMR (CDCl3,
100 MHz): d: 12.1; 18.6; 18.8; 21.7; 22.7; 23.9; 25.4; 25.6;
26.3; 28.1; 28.2; 29.5; 31.7; 35.9; 36.3; 37.0; 37.4; 39.6; 39.7;
40.0; 40.7; 40.8; 43.0; 46.1; 46.6; 46.8; 47.6; 54.2; 56.3; 59.0;
71.5; 122.8; 139.0. IE-SM: m=z ð%Þ ¼ 471ð1Þ; 414 (1); 401
20
D
(19); 400 (33); 385 (50); 384 (100); 369 (11); 351 (26). ½aꢀ
þ41 (c 0.2 M, MeOH). Anal. Calcd for C34H61N3O: C
77.35; H 11.66; N 7.96. Found: C 77.31; H 11.69; N 7.95.
20. The anti-microbial tests were measured in vitro using a
liquid-phase turbidimetric system (Bioscreenâ from Lab-
system, France) and automatically evaluated every 30 min
for 48 h. The tested molecules were dissolved in dimeth-
ylsulfoxide and added in liquid broth to prepare various
concentrations of drugs by successives dilutions (DMSO
in liquid broth did not exceed 2%). The inocula (2%) was
prepared from overnight culture and diluted to obtain 0.5
value of optical density (600 nm). Microorganisms were
incubate in shaker system and the growth curves were
obtained by optical density readings with wide broad filter
(420–580 nm) of Bioscreen system Dei-Cas, E.; Dujardin,
L.; Ribiero Pinto, M. E.; Ajana, F.; Fruit, J.; Poulain, D.;
Camus, D.; Vernes, A. Mycoses 1991, 34, 167.
16. Dherbomez, M.; El Kihel, L.; Letourneux, Y. FEMS
Microbiol. Lett. 1995, 126, 91.
17. El Kihel, L.; Choucair, B.; Dherbomez, M.; Letourneux,
Y. Eur. J. Org. Chem. 2002, 4075.
18. El Kihel, L.; Bosch, S.; Dherbomez, M.; Roussakis, C.;
Letourneux, Y. Anticancer Res. 1999, 19, 1229.
19. Compound I: 1H NMR (CDCl3, 400 MHz): d: 0.68 (s, 3H,
Me 18); 0.86 (d, 6H, J ¼ 6:5, Me 27 and Me 26); 0.91 (d,
3H, J ¼ 6:5, Me 21); 0.99 (s, 3H, Me 19); 2.66 (m, 1H, St–
21. Cell line and cell culture: the NSCLC-N6 cell line, derived
from a human non-small cell bronchopulmonary carci-
noma (moderately differentiated, rarely keratinizing,
classified as T2NOMO) was used for all experiments.
The cells were cultured in RPMI 1640 medium with 5%
foetal calf serum, to which were added 100 IU penicil-
lin mLꢁ1, 100 lg streptomycin mLꢁ1 and 2 mM glutamine,
at 37 °C in a air/carbon dioxide (95:5, v/v) atmosphere. In
these conditions, cell doubling time was 48 h. Cells used in
all experiments never exceeded 35 passages. Cytotoxicity
determinations: continuous drug exposure. Experiments
were performed in 96 wells microliter plates
(2 ꢂ 105 cells mLꢁ1). Cell growth was estimated by a
colorimetric assay based on the conservation of tetrazo-
lium dye (MTT) to a blue formazan product by live
mitochondria. Eight repeats were performed for each
concentration. Control growth was estimated from 16
determinations. Optical density at 570 nm corresponding
to solubilized formazan was read for each well on a
Titertek Multiskan MKII Mossmann, T. J. Immunol.
Methods 1983, 65, 55.
0
NH–CHaHa –(CH2)2–NH–); 2.74 (dd, 1H, J7b–8 ¼ 4:3,
J7b–6 ¼ 5:0, H7b of a epimer); 2.80 (t, 2H, J ¼ 5:4,
0
–HN–(CH2)3–CH2–NH2); 2.92 (m, 1H, St–NH–CHaHa –
(CH2)2–NH–); 3.00–3.48 (m, 4H, St–NH–(CH2)2–CH2–
NH– et NH–CH2–(CH2)3–NH2); 3.63 (m, 1H, H3a); 5.66
(dd, 1H, J6–7b ¼ 1:8, J6–4 < 1:0, H6 of a epimer); 7.23 (br s,
2H, –NH2, D2O exchange); 8.15 (br s, 1H, –NH–, D2O
exchange); 8.44 (br s, 1H, –NH–, D2O exchange). 13C
NMR (CDCl3, 100 MHz): d: 12.1; 18.6; 18.8; 21.7; 22.7;
23.9; 25.4; 25.6; 26.3; 28.1; 28.2; 29.5; 31.7; 35.9; 36.3; 37.0;
37.4; 39.6; 39.7; 40.0; 40.7; 40.8; 43.0; 46.1; 46.36; 46.8;
47.6; 54.2; 56.3; 65.4; 71.5; 122.2; 139. IE-SM:
m=z ð%Þ ¼ 471 (3); 457 (2); 442 (2); 414 (2); 401 (35);
400 (100); 385 (24); 384 (27); 369 (3); 351 (24); 275 (10).
20
½aꢀ ꢁ41 (c 0.2 M, MeOH). Anal. Calcd for C34H61N3O: C
D
77.35; H 11.66; N 7.96. Found: C 77.33; H 11.69; N 7.95.
Compound II: 1H NMR (CDCl3, 400 MHz): d: 0.69 (s,