+
◦
3
6
(
(
0.4, 32.3, 33.0, 53.1 and 53.6 (2 × N CH
3
), 61.2 (CH
2
OH),
(3 mL) in a sealed tube at 90 C for 24 h. The solvent was removed
5.1, 66.7, 68.7, 69.3, 70.1 (signal overlap), 70.5, 72.0, 72.6, 73.5
in vacuo and the residue triturated at low temperature (diethyl
−
1
CHOCH
+ES) 797 (M − Br, 100%); Found (+HRFAB) (M − Br),
96.7399. C49 requires 796.7394; Found C, 66.19; H, 10.99;
N, 1.50. C49 O requires C, 65.74; H, 11.26; N, 1.56%.
Br·H
2
), 129.8 (2 × CH=CH), 130.0 (2 × CH=CH); m/z
ether) to obtain 37 (73 mg, 67%). mmax (film)/cm 2922, 2853,
1462; d (300 MHz; CDCl CH ),
) 0.85 (6H, t, J 6.6, 2 × CH
1.24 (44H, m), 1.51 (4H, m, 2 × OCH CH CH ), 1.98 (8H, m,
2 × CH ), 3.40–4.14 (32H, m, 2 × CH O (PEG),
CH=CHCH
CH OCH CH , CHOCH , CHOCH
, 3 × N CH
), 5.35 (4H, m, 2 × CH=CH); d
(75 MHz; CDCl
CH
+
+
H
3
2
3
7
H
98NO
6
2
2
2
H
98NO
6
2
2
2
2
+
2
OC18
H
35, CH
2
2
2
2
2
2
3
,
)
2
,3-Di-((9Z)-octadecenyloxy)propyl-N -(2-{2-[2-(2-hydroxy-
ethoxy)ethoxy]ethoxy}ethyl)-N,N-dimethylammonium chloride
17b). Crude 17a (approximately 250 mg) (generated as
described above) in methanol (1 mL) was passed through an
Amberlite IRA-400 (Cl) ion exchange column eluting with
dichloromethane–methanol (1 : 1). The solvents were removed
in vacuo and the crude product purified by low temperature
recrystallisation (ethyl acetate) to yield 17b as a pale yellow oil
+
5
1
× N CH
3
C
4.1 (2 × CH
2
3
), 22.7, 26.1, 26.2, 27.2, 29.2–29.8 (signal
(
+
+
overlap), 30.1, 31.9, 32.6, 52.9 (N CH
3
), 53.8 (N CH
3
), 54.7
+
(
3 × N CH
29.8 (2 × CH=CH), 130.0 (2 × CH=CH); m/z (+ES) 375.6
[M − 2Br] , 100%); Found (+HRFAB) (M − Br), 829.6729.
3
), 64.8, 65.0, 65.5, 65.9, 66.6, 69.0, 69.4, 72.1, 73.4,
ꢀ
R
1
1
+
+
79
(
2
C
48
H
98BrN
2
O requires 829.6755.
3
(
∼90% from crude 17a).
,3-Di-((9Z)-octadecenyloxy)propyl-N-[2-(2-bromoethoxy)ethyl]-
N,N-dimethylammonium bromide (31). 2-Bromoethyl ether (25)
2
,3-Di-((9Z)-octadecenyloxy)propyl-N -3-(N,N,N -trimethyl-
ammoniumpropyl)-N,N-dimethylammonium dibromide (40).
Bromide 34 (100 mg, 0.12 mmol) and trimethylamine (45 wt% in
2
H
2
O; 0.094 mL, 0.61 mmol) were stirred in methanol (2 mL) in a
(
278 mg, 1.20 mmol) and the amine 6 (300 mg, 0.483 mmol) were
◦
◦
sealed tube at 90 C for 24 h. The solvent was removed in vacuo
and the product purified by low temperature recrystallisation
stirred in methanol (2 mL) at 90 C in a sealed tube for 24 h. The
solvent was removed in vacuo and the product purified by low
temperature recrystallisation (ethyl acetate) to yield 31 as a pale
−
1
(
ethyl acetate) to give 40 (87 mg, 82%). mmax (CHCl
2852, 1656, 1456; d (300 MHz; CDCl
) 0.86 (6H, t, J 6.5, 2 ×
CH CH CH ), 2.00 (8H,
), 1.25 (46H, m), 1.53 (4H, m, 2 × OCH
m, 2 × CH ), 3.38–4.01 (27H, m, CH
CH=CHCH
CH OCH CH , CHOCH
m, CHOCH
), 5.33 (4H, m, 2 × CH=CH); d
14.1 (2 × CH CH ), 22.6, 25.8, 26.0, 27.1, 29.0–29.6 (signal
overlap), 29.9, 31.7, 32.4, 51.5 (N CH
3
)/cm 2922,
−
1
H
3
yellow oil (197 mg, 48%). mmax (film)/cm 2927, 2854, 2344, 1642,
2
3
2
2
1
1
2
3
2
d
2
3
465; d
.25 (44H, m), 1.52 (4H, m, 2 × OCH
× CH ), 3.41 (4H, m, CH
CH=CHCH
O (PEG), CHOCH
H
(300 MHz; CDCl
3
) 0.84 (6H, t, J 7.0, 2 × CH
CH CH ), 1.98 (8H, m,
OCH CH , CHOCH ),
, CH
), 5.31 (4H, m, 2 × CH=CH);
) 14.0 (2 × CH CH ), 22.6, 26.0, 26.2, 27.2,
2
CH
3
),
2
2
2
OC18
), 4.11 (1H,
(75 MHz; CDCl )
H ,
35
2
2
2
+
+
2
2
2
2
, 3 × N CH
2
, 5 × N CH
3
2
2
2
2
2
2
2
C
3
.48–4.10 (19H, m, 2 × CH
2
2
2
OC18
H ,
35
+
+
2
3
× N CH
2
, CH
(75 MHz; CDCl
2
Br, 2 × N CH
3
+
+
3
), 53.6 (N CH
3
), 54.0 (3 ×
), 129.6
C
3
2
3
+
N CH
3
), 62.6, 63.1, 67.2, 68.8, 69.2, 71.9, 73.0 (CHOCH
2
9.1, 29.2–29.4 (signal overlap), 29.5, 29.6, 29.7, 30.0, 30.8, 31.8,
1
+
(2 × CH=CH), 129.7 (2 × CH=CH); m/z (+ES) 361 ( [M −
2.5, 53.3 and 53.5 (2 × N CH
3
), 64.9, 66.8, 68.6, 69.3, 71.0, 72.0
), 129.7 (2 × CH=CH), 129.9
2
+
+
2
Br] , 100%); Found (+HRFAB) (MNa − 2Br) , 743.7350.
47 96 2 2
(
(
(
signal overlap), 73.4 (CHOCH
2
+
81
C H O N Na requires 743.7346.
2 × CH=CH); m/z (+ES) 773 (M − Br ( Br), 100%), 771
M − Br ( Br), 88); Found (+HRFAB) (M − Br), 770.6039.
NBr requires 770.6020.
+
79
+
C
45
H
89
O
3
Acknowledgements
2
,3-Di-((9Z)-octadecenyloxy)propyl-N-(3-bromopropyl)-N,N-
The EPSRC are thanked for a DTA studentship to C.A.H., J.B.W.,
and an EPSRC-LSI grant to J.H. (GR/S05878/01). M.W was
supported by a grant from the BBSRC. The authors also thanks
Dieter Gruenert, California Pacific Medical Center Research
Insitiute, San Francisco, CA, for kindly providing the cell lines
dimethylammonium bromide (34). Amine 6 (0.500 g, 0.806 mmol)
and 1,3-dibromopropane (28) (0.82 mL, 8.10 mmol) were stirred
in hexane (2 mL) in a sealed tube at 80 C for 18 h. The solvent
◦
was removed in vacuo and the product purified by flash silica gel
chromatography (gradient: CH
2
Cl
2
to 10% MeOH in CH
= 0.17 (5% MeOH
); mmax (film)/cm 2924, 2853, 1634, 1464; d (300 MHz;
) 0.85 (6H, t, J 6.7, 2 × CH CH ), 1.27 (44H, m), 1.53 (4H,
CH CH ), 1.97 (8H, m, 2 × CH ), 2.42
CH=CHCH
CH Br), 3.40 (4H, m, CH OCH CH , CHOCH ),
.45–3.92 (14H, m, CH , CH Br, 2 ×
35, 2 × N CH
), 5.32 (4H, m, 2 × CH=CH); d
CH ), 22.7, 26.1, 26.3, 27.3, 28.7,
2
Cl
2
) to
1
HAEo− and SCFTE29o−.
yield 34 as a pale yellow oil (0.410 g, 62%). R
f
−
1
in CH
CDCl
2
Cl
2
H
References
3
2
3
m, 2 × OCH
2
2
2
2
2
1
N. Somia and I. M. Verma, Nat. Rev. Genet., 2000, 1, 91–99.
(
3
2H, m, CH
2
2
2
2
2
2
2 A. D. Miller, Angew. Chem., Int. Ed., 1998, 37, 1768–1785.
3 M. E. Davis, Curr. Opin. Biotechnol., 2002, 13, 128–131.
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2
OC18
H
2
2
4
5
K. Kostarelos and A. D. Miller, Chem. Soc. Rev., 2005, 34, 970–994.
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+
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3
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2
C
(
2
75 MHz; CDCl
3
) 14.0 (2 × CH
2
3
6 C. M. Varga, T. J. Wickham and D. A. Lauffenburger, Biotechnol.
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9.2, 29.3–29.8 (signal overlap), 30.1, 31.9, 32.6, 52.8 and 53.0 (2 ×
+
7 For exampleT. Tagawa, M. Manvell, N. Brown, M. Keller, E. Perouzel,
K. D. Murray, R. P. Harbottle, M. Tecle, F. Booy, M. C. Brahimi-Horn,
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Ther., 2002, 9, 564–576.
N CH
3
), 64.5, 65.8, 68.4, 69.4, 72.1, 73.3 (CHOCH
2
), 129.7 (2 ×
+ 81
CH=CH), 129.8 (2 × CH=CH); m/z (+ES) 743 (M − Br ( Br),
+
79
+
79
1
7
00%), 741 (M − Br ( Br), 88); Found (+HRFAB) (M − Br),
40.5939. C44 requires 740.5915.
8
S. L. Hart, C. V. Arancibia-C a´ rcamo, M. A. Wolfert, C. Mailhos, N. J.
O’Reilly, R. R. Ali, C. Coutelle, A. J. T. George, R. P. Harbottle, A. M.
Knight, D. F. P. Larkin, R. J. Levinsky, L. W. Seymour, A. J. Thrasher
and C. Kinnon, Hum. Gene Ther., 1998, 9, 575–585.
P. L. Felgner, T. R. Gadek, M. Holm, R. Roman, H. S. Chan, M. Wenz,
J. P. Northrop, G. M. Ringold and H. Danielsen, Proc. Natl. Acad. Sci.
U. S. A., 1987, 84, 7413–7417.
H
87BrNO
2
2
,3-Di-((9Z)-octadecenyloxy)propyl-N-[2-(2-(N,N,N-trimethyl-
ammonium)ethoxy)ethyl]-N,N-dimethylammonium dibromide
37). Bromide 31 (100 mg, 0.118 mmol) and trimethylamine
45 wt% in H O; 0.36 mL, 2.35 mmol) were stirred in methanol
9
(
(
2
2
558 | Org. Biomol. Chem., 2008, 6, 2554–2559
This journal is © The Royal Society of Chemistry 2008