894
T. Ren et al. / Bioorg. Med. Chem. Lett. 10 (2000) 891±894
Table 1. Transfection activity of vitamin D-based cationic lipids in dierent cell lines
Cell line
RLU/wella
1a
1b
1c
1d
2a
2b
2c
2d
3LL
8.0 Â 105
2.7 Â 105
2.6 Â 106
3.9 Â 106
4.0 Â 103
1.8 Â 105
3.8 Â 104
1.6 Â 106
6.1 Â 105
2.1 Â 106
9.3 Â 105
6.3 Â 107
5.9 Â 105
1.9 Â 106
5.4 Â 105
4.8 Â 107
2.2 Â 105
1.2 Â 105
2.9 Â 105
1.1 Â 106
5.3 Â 103
5.7 Â 105
1.8 Â 104
5.6 Â 105
8.2 Â 105
7.8 Â 106
1.7 Â 106
4.7 Â 107
1.5 Â 105
6.9 Â 105
1.4 Â 106
3.2 Â 107
NIH3T3
HeLa
293
aDierent cells (5 Â 104) were seeded in 48-well plate 24 h before the addition of DNA/lipid complexes (1 mg DNA/5 nmol cationic lipid per well).
The level of luciferase gene expression is expressed as mean of relative light units (RLU) per well of triplicate experiments (n=3).
In conclusion, we have synthesized a new panel of
steroidal cationic lipids and evaluated their in vitro
transfection activity. Among the lipids tested, poly-
amines 1d, 2d exhibited superior transfection activity in
a broader range of lipid to DNA ratios in BL-6 cells.
Transfection results obtained from the additional cell
lines suggest that these new vitamin D-based cationic
lipids are useful transfection reagents for in vitro gene
transfer studies.
A. J.; Eaton, M. A. W.; Baker, T.; Catterall, C.; Blagbrough,
I. S. FEBS Lett. 1999, 459, 337.
8. Walker, S.; So®a, M. J.; Kakarla, R.; Kogan, N. A.; Wierichs,
L.; Longley, C. B.; Bruker, K.; Axelrod, H. R.; Midha, S.;
Babu, S.; Kahne, D. Proc. Natl. Acad. Sci. USA 1996, 93, 1585.
9. (a) Ren, T.; Liu, D. Tetrahedron Lett. 1999, 40, 7621. (b)
Ren, T.; Liu, D. Tetrahedron Lett. 1999, 40, 209. (c) Ren, T.;
Liu, D. Bioorg. Med. Chem. Lett. 1999, 9, 1247. (d) Ren, T.;
Zhang, G.; Song, Y. K.; Liu, D. J. Drug Targeting 1999, 7,
285.
10. Hormones, 2nd ed.; Norman, A. W.; Litwack, G., Eds.;
Academic: San Diego, CA, 1997.
1
11. Compound 1: H NMR (300 MHz, CDCl3) d 8.11 (s, 1H,
Acknowledgements
imidazole ring), 7.49 (s, 1H, imidazole ring), 7.06 (s, 1H, imi-
dazole ring), 6.25 (d, J=11.2 Hz, 1H), 6.02 (d, J=11.2 Hz,
1H), 5.20 (m, 1H), 5.19 (m, 2H), 5.11(s, 1H), 4.89 (s, 1H),
2.86±2.65 (m, 2H), 2.60±2.40 (m, 2H), 2.30 (m, 1H), 2.20±1.80
(m, 5H), 1.70±1.60 (m, 4H), 1.60±1.40 (m, 4H), 1.40±1.20 (m,
3H), 1.02 (d, J=6.6 Hz, 3H), 0.91 (d, J=7 Hz, 3H), 0.83 (d,
J=6.6 Hz, 3H), 0.82 (d, J=6.6 Hz, 3H), 0.56 (s, 3H). Com-
pound 2: 1H NMR (300 MHz, CDCl3) d 8.12 (s, 1H, imidazole
ring), 7.41 (s, 1H, imidazole ring), 7.06 (s, 1H, imidazole ring),
6.26 (d, J=11.2 Hz, 1H), 6.03 (d, J=11.2 Hz, 1H), 5.20 (m,
1H), 5.12 (s, 1H), 4.90 (s, 1H), 2.85±2.70 (m, 2H), 2.60±2.40
(m, 2H), 2.30 (m, 1H), 2.20±1.80 (m, 5H), 1.80±1.60 (m, 3H),
1.60±1.40 (m, 4H), 1.40±1.20 (m, 6H), 1.20±1.05 (m, 3H), 0.92
(d, J=6 Hz, 3H), 0.87 (d, J=6.6 Hz, 6H), 0.55 (s, 3H). Com-
pound 1b: 1H NMR (300 MHz, CDCl3) d 6.20 (d, J=11.2 Hz,
1H), 6.02 (m, 1H), 6.01(d, J=11.2 Hz, 1H), 5.19 (m, 2H), 5.05
(s, 1H), 4.85 (s, 1H), 4.82 (m, 1H), 3.83 (m, 2H), 3.79 (m, 2H),
3.47 (s, 9H, N+Me3), 2.80 (m, 1H), 2.57 (m, 1H), 2.35 (m,
1H), 2.19 (m, 1H), 2.10±1.80 (m, 6H), 1.70 (m, 4H), 1.50 (m,
4H), 1.30 (m, 3H), 1.01 (d, J=6.6 Hz, 3H), 0.91 (d, J=6.7 Hz,
3H), 0.83 (d, J=6.6 Hz, 3H), 0.82 (d, J=6.6 Hz, 3H), 0.55 (s,
Assistance from Professor Joseph E. Knapp for critical
reading of this manuscript is acknowledged. This work
was supported in part by a grant from NIH (CA 72925)
and research contract with Targeted Genetics Corporation.
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2H), 2.20 (m, 1H), 2.00±1.80 (m, 5H), 1.70±1.60 (m, 3H), 1.60±
1.45 (m, 4H), 1.40±1.20 (m, 6H), 1.20±1.10 (m, 3H), 0.92 (d,
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