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M. Lukác et al. / European Journal of Medicinal Chemistry 66 (2013) 46e55
53
2.05e2.17 (m, 2H, NþCH2CH2CH2O), 3.30 (s, 9H, Nþ(CH3)3), 3.60e
3.84 (m, 4H, NþCH2CH2CH2O and OCH2CH2), 3.90e4.01 (m,
7.19 (d, J ¼ 8.8 Hz, 2H, Hortho(Ph)), 7.29e7.39 (m, 3H, Hmeta(Bn) and
Hpara(Bn)), 7.50 (d, J ¼ 7.7 Hz, 2H, Hortho(Bn)); 13C NMR (CDCl3)
d: 31.6,
2H, NþCH2CH2CH2O); 13C NMR (CDCl3)
d
: 14.1, 22.7, 24.8, 26.0,
31.7, 32.3, 37.9, 50.4, 56.8, 59.2, 63.7, 64.2, 68.0, 69.0, 113.7, 127.1,
29.4, 29.6, 29.7, 29.8, 31.1, 31.9, 53.3, 61.6, 64.1, 65.3; 31P NMR
127.7, 129.1, 130.4, 133.3, 142.4, 156.3; 31P NMR (CDCl3)
d
: ꢁ0.71; IR
(CDCl3)
d
: ꢁ0.33; IR ymax/cmꢁ1 3400, 2917, 2850, 1660, 1493, 1470,
ymax/cmꢁ1 3348, 2954, 2892, 1632, 1611, 1512, 1484, 1251, 1080,
1241, 1053, 943, 837, 719; HRMS calcd. for C22H48O4NPNa ¼
1059, 985, 795, 763, 710, 541; HRMS calcd. for
444.3213; found m/z: 444.3209 [M þ Na]þ.
C
27H42O5NPNa ¼ 514.2693; found m/z: 514.2685 [M þ Na]þ.
4.1.6.2. 2-[Benzyl(dimethyl)ammonio]ethyl hexadecyl phosphate
4.1.6.7. 3-[Benzyl(dimethyl)ammonio]propyl 2-[(2,2,4,4-
(HPC2Bn). Yield 58%; 1H NMR (CDCl3)
d: 0.88 (t, J ¼ 6.6 Hz, 3H,
tetramethylpentane-2-yl)oxy]ethyl phosphate (PC3Bzt). Yield 24%;
CH2CH3), 1.25 (br s, 26H, OCH2CH2(CH2)13CH3), 1.50e1.61 (m, 2H,
OCH2CH2), 3.30 (s, 6H, Nþ(CH3)2), 3.79e3.98 (m, 4H, NþCH2CH2O
and OCH2CH2), 4.34e4.43 (m, 2H, NþCH2CH2O), 4.89 (s, 2H, CH2Ph),
7.36e7.46 (m, 3H, Hmeta and Hpara), 7.68 (d, J ¼ 6.6 Hz, 2H, Hortho);
1H NMR (CDCl3)
d: 0.68 (s, 9H, (CH3)3CCH2(CH3)2C), 1.30 (s, 6H,
(CH3)3CCH2(CH3)2C), 1.67 (s, 2H, (CH3)3CCH2(CH3)2C), 2.18e2.31 (m,
2H, NþCH2CH2CH2O), 3.01 (s, 6H, Nþ(CH3)2), 3.75e3.82 (m, 2H,
NþCH2CH2CH2O), 4.01e4.16 (m, 4H, PhOCH2CH2O and
NþCH2CH2CH2O), 4.18e4.22 (m, 2H, PhOCH2CH2O), 4.67 (s, 2H,
CH2Ph), 6.74 (d, J ¼ 8.7 Hz, 2H, Hmeta(Ph)), 7.19 (d, J ¼ 8.7 Hz, 2H,
Hortho(Ph)), 7.31e7.42 (m, 3H, Hmeta(Bn) and Hpara(Bn)), 7.51 (d,
13C NMR (CDCl3)
d: 14.2, 22.8, 26.0, 29.5, 29.6, 29.7, 29.8, 31.1, 31.3,
32.0, 50.5, 59.2, 64.1, 65.7, 69.0, 127.8, 129.1, 130.6, 133.6; 31P NMR
(CDCl3)
d
: 0.31; IR ymax/cmꢁ1 3443, 2920, 2851, 1632, 1469, 1241,
1068, 988, 928, 762, 719; HRMS calcd. for C27H50O4NPNa ¼
J ¼ 7.7 Hz, 2H, Hortho(Bn)); 13C NMR (CDCl3)
d: 24.6, 31.7, 31.8, 32.3,
506.3370; found m/z: 506.3360 [M þ Na]þ.
37.9, 49.3, 56.9, 61.8, 63.0,63.8, 67.7, 68.1, 113.6, 127.1, 127.7, 129.1,
130.4, 133.3, 142.3, 156.4; 31P NMR (CDCl3)
d
: ꢁ1.08; IR ymax/cmꢁ1
4.1.6.3. 2-[Benzyl(dimethyl)ammonio]propyl hexadecyl phosphate
3427, 2956, 2886, 1632, 1610, 1511, 1480, 1248, 1096, 1067, 961, 832,
768, 735, 706, 534; HRMS calcd. for C28H44O5NPNa ¼ 528.2850;
found m/z: 528.2844 [M þ Na]þ.
(HPC3Bn). Yield: 70%; 1H NMR (CDCl3)
d
: 0.88 (t, J ¼ 6.7 Hz, 3H,
CH2CH3), 1.25 (br s, 26H, OCH2CH2(CH2)13CH3), 1.43e1.62 (m, 2H,
OCH2CH2), 2.18e2.38 (m, 2H, NþCH2CH2CH2O), 3.14 (s, 6H, Nþ(CH3)2),
3.62e3.82 (m, 4H, NþCH2CH2CH2O and OCH2CH2), 3.85e4.11 (m, 2H,
NþCH2CH2CH2O), 4.80 (s, 2H, CH2Ph), 7.28e7.43 (m, 3H, Hmeta and
4.2. Equilibrium surface tension
Hpara), 7.45e7.67 (m, 2H, Hortho); 13C NMR (CDCl3)
d: 14.2, 22.8, 24.7,
The critical micelle concentration values of the surfactants were
determined from the surface tension isotherm. The solvent surface
tension values were measured by the Wilhelmy plate technique
using a Kruss 100 MK2 tensiometer. Deionized water was used in the
preparation of all samples. The temperature of the measurements
was kept at 25 ꢃ 0.1 ꢂC. Measurements of equilibrium surface ten-
sion were taken repeatedly (every 6 min) until the change in surface
tension was less than 0.05 mN mꢁ1. (The measurement of one so-
lution with premicellar concentrations of the compounds took 1e
2 h, whereas the measurement of the equilibrium surface tension of
premicellar solutions of HPC2Bn and HPC3Bn took 1.5e3 h.) The
critical micelle concentrations (cmc) and surface tensions at the cmc
26.1, 29.4, 29.6, 29.8, 31.2, 31.3, 32.0, 49.5, 61.7, 62.5, 65.4, 67.5, 128.0,
129.1,130.4,133.5; 31PNMR (CDCl3)
d
:ꢁ0.53; IR ymax/cmꢁ13414, 2920,
2851,1632,1487,1468,1240,1094,1068, 961, 834, 777, 732, 704; HRMS
calcd. for C28H52O4NPNa ¼
520.3526; found m/z: 520.3522 [M þ Na]þ.
4.1.6.4. Hexadecyl 2-pyridinium-1-ylethyl phosphate (HPC2P).
Yield 42%; 1H NMR (CDCl3)
d
: 0.88 (t, J ¼ 6.7 Hz, 3H, CH2CH3), 1.25
(br s, 26H, OCH2CH2(CH2)13CH3), 1.43e1.54 (m, 2H, OCH2CH2),
3.62e3,73 (m, 2H, OCH2CH2), 4.22e4.38 (m, 2H, NþCH2CH2O),
4.91e5.08 (m, 2H, NþCH2CH2O), 8.03 (t, J ¼ 6.7 Hz, 2H, Hmeta), 8.42
(t, J ¼ 7.9 Hz, 1H, Hpara), 9.25 (d, J ¼ 5.6 Hz, 2H, Hortho); 13C NMR
(gcmc) were determined from the intersection of two lines e one
(CDCl3) d: 14.1, 22.7, 25.9, 29.4, 29.5, 29.6, 29.7, 29.8, 31.0, 31.1, 32.0,
premicellar and second one postmicellar e of the surface tension vs.
log c curve. From the surface tension data, the adsorbed amount of
surfactant Gcmc is calculated utilizing the Gibbs adsorption isotherm
62.2, 63.8, 65.6, 128.1, 145.2, 146.1; 31P NMR (CDCl3)
d: 0.55; IR
ymax/cmꢁ1 3382, 2917, 2850,1690,1637,1493,1471,1239,1218,1079,
1048, 1028, 918, 790, 782, 746, 719, 685; HRMS calcd. for
C
23H42O4NPNa ¼ 450.2744; found m/z: 450.2734 [M þ Na]þ.
Gcmc ¼ ꢁ½d
g
=dlog cꢄT =ð2:303iRTÞ
(1)
where
g
is the surface tension (mN mꢁ1), c is the surfactant con-
4.1.6.5. Hexadecyl 3-pyridinium-1-ylpropyl phosphate (HPC3P).
Yield 35%; 1H NMR (CDCl3)
centration, i is the prefactor (QUATs, i ¼ 2; APCs, i ¼ 1), T is the
absolute temperature and R the gas constant. Surface excess may be
determined from the slope below the cmc in the surface tension vs.
log c plots. Surface area at the surface saturation per head group
(Acmc) is obtained from the equation
d
: 0.88 (t, J ¼ 6.7 Hz, 3H, CH2CH3), 1.25
(br s, 26H, OCH2CH2(CH2)13CH3), 1.50e1.63 (m, 2H, OCH2CH2),
2.23e2.36 (m, 2H, NþCH2CH2CH2O), 3.75e3.81 (m, 2H, OCH2CH2),
3.81e3.92 (m, 2H, NþCH2CH2CH2O), 5.03 (t, J ¼ 6.2 Hz, 2H,
NþCH2CH2CH2O), 8.07 (t, J ¼ 7.0 Hz, 2H, Hmeta), 8.39 (t, J ¼ 7.2 Hz,1H,
Hpara), 9.52 (d, J ¼ 5.9 Hz, 2H, Hortho); 13C NMR (CDCl3)
d: 14.1, 22.7,
Acmc ¼ 1020=NAGcmc
(2)
26.0, 29.4, 29.5, 29.6, 29.7, 29.9, 31.0, 31.9, 33.6, 58.9, 60.5, 65.5,
128.3, 144.6, 146.1; 31P NMR (CDCl3) : 0.80; IR ymax/cmꢁ1 3298,
d
were NA is the Avogadro constant.
2915, 2849, 1636, 1488, 1471, 1255, 1233, 1119, 1093, 1045, 1025,
1009, 975, 831, 814, 717, 675; HRMS calcd. for C24H44O4NPNa ¼
464.2900; found m/z: 464.2892 [M þ Na]þ.
4.3. Biological activities
4.3.1. Cytotoxicity assay
4.1.6.6. 2-[Benzyl(dimethyl)ammonio]ethyl 2-[(2,2,4,4-
tetramethylpentane-2-yl)oxy]ethyl phosphate (PC2Bzt). Yield 22%;
4.3.1.1. Cell culture. The following human cancer cell lines were used
for this study: CEM (acute T-lymphoblastic leukemia), HeLa (cervical
carcinoma cells), A-549 (human lung adenocarcinoma), MDA-MB-
231 and MCF-7 (breast cancer cells). CEM and HeLa cells were
maintained in RPMI 1640 medium (PAA Laboratories, Pasching,
Austria). MDA-MB-231, MCF-7 and A-549 were maintained in growth
1H NMR (CDCl3)
d: 0.68 (s, 9H, (CH3)3CCH2(CH3)2C), 1.29 (s, 6H,
(CH3)3CCH2(CH3)2C), 1.66 (s, 2H, (CH3)3CCH2(CH3)2C), 3.19 (s, 6H,
Nþ(CH3)2), 3.81e3.88 (m, 2H, NþCH2CH2O), 4.06e4.14 (m, 2H,
PhOCH2CH2O), 4.18e4.26 (m, 2H, PhOCH2CH2O), 4.42e4.52 (m, 2H,
NþCH2CH2O), 4.75 (s, 2H, CH2Ph), 6.72 (d, J ¼ 8.8 Hz, 2H, Hmeta(Ph)),