Arch. Pharm. Chem. Life Sci. 2005, 338, 299−304
Isosteric Analogues of the HIV Drug GCA-186
303
1-(Allyloxymethyl)-6-(3,5-dichlorobenzyl)-5-ethyluracil (6b)
Yield 304 mg (98%); mp 115Ϫ117°C. 1H-NMR (CDCl3):
6-(3,5-Dichlorobenzyl)-1-(prop-2-ynyloxymethyl)-5-ethyluracil (6g)
Yield 81 mg (26%); mp 147Ϫ151°C. 1H-NMR (CDCl3): δ (ppm) ϭ
1.08 (t, 3H, J ϭ 7.4 Hz, CH3), 2.45 (q, 2H, J ϭ 7.3 Hz, CH2),
2.4Ϫ2.5 (m, 1H, sCH), 4.12 (s, 2H, CH2), 4.29 (d, 2H, J ϭ 2.3 Hz,
CH2), 5.18 (s, 2H, CH2), 7.03Ϫ7.30 (m, 3H, aryl), 9.62 (s, 1H, NH).
13C-NMR (CDCl3): δ (ppm) ϭ 13.73 (CH3), 19.19 (CH2), 32.79
(CH2), 57.20 (CH2), 72.56 (CH), 74.98 (CH2), 78.86 (Cϭ), 117.72
(C-5), 125.84, 127.84, 135.91, 138.47 (aryl), 147.09 (C-2), 151.72 (C-
6), 163.01 (C-4). Anal. calcd. for C17H16Cl2N2O3 (367.23): C, 55.60;
H, 4.39; N, 7.63. Found: C, 55.64; H, 4.40; N, 7.37.
δ
(ppm) ϭ 1.08 (t, 3H, J ϭ 7.2 Hz, CH3), 2.44 (q, 2H, J ϭ 7.3 Hz,
CH2), 4.10Ϫ4.13 (m, 4H, 2 ϫ CH2), 5.14 (s, 2H, CH2), 5.20Ϫ5.32
(m, 2H, CϭCH2), 5.86 (m, 1H, CHϭC), 7.01Ϫ7.29 (m, 3H, aryl),
9.71 (s, 1H, NH). 13C-NMR (CDCl3): δ (ppm) ϭ 13.69 (CH3), 19.18
(CH2), 32.84 (CH2), 70.59 (CH2), 72.58 (CH2), 117.59 (C-5), 118.05
(ϭCH2), 125.78, 127.75, 135.88, 138.64 (aryl), 133.28 (CHϭ),
147.23 (C-2), 151.73 (C-6), 163.11 (C-4). Anal. calcd. for
C17H18Cl2N2O3 (369.25): C, 55.30; H, 4.91; N, 7.59. Found: C,
55.53; H, 4.91; N, 7.59.
6-(3,5-Dichlorobenzyl)-5-ethyl-1-(indan-1-yloxy)methyl-uracil (6h)
Yield, 232 mg (62%) as a white foam. 1H-NMR (CDCl3): δ
(ppm) ϭ1.05 (t, 3H, J ϭ 7.4 Hz, CH3), 1.97Ϫ2.07 (m, 1H, 2Ј-H),
2.32Ϫ2.48 (m, 3H, 2Ј-H, CH2), 2.77Ϫ2.87 (m, 1H, 3Ј-H), 3.03Ϫ3.12
(m, 1H, 3’-H), 4.07 (s, 2H, CH2), 5.08Ϫ5.37 (m, 3H, 1Ј-H, CH2),
6.91 (s, 2H, Harom.), 7.18Ϫ7.35 (m. 5H, Harom.), 9.45 (s, 1H, NH).
13C-NMR (CDCl3): d(ppm) ϭ13.79 (CH3), 19.21 (CH2), 30.15 (C-
2’), 32.75 (C-3’), 32.83 (CH2), 71.59 (CH2), 82.40 (C-1’), 117.60 (C-
5), 124.86, 125.04, 125.77, 126.56, 127.73, 128.88, 135.83, 138.59,
141.58, 144.00 (Carom.), 147.41 (C-6), 151.61 (C-2), 162.99 (C-4).
HRMS-MALDI: m/z ϭ 467.0900 (M ϩ Naϩ, C23H22Cl2N2NaO3);
requires 467.0899.
6-(3,5-Dichlorobenzyl)-1-(3-methylbut-2-enyloxymethyl)-5-ethyl-
uracil (6c)
Yield 299 mg (90%); mp 103Ϫ105°C. 1H-NMR (CDCl3):
δ
(ppm) ϭ 1.09 (t, 3H, J ϭ 7.4 Hz, CH3), 1.66 (s, 3H, CH3), 1.75 (s,
3H, CH3), 2.43 (q, 2H, J ϭ 7.4 Hz, CH2), 4.10 (d, 2H, J ϭ 6.9 Hz,
CH2), 4.12 (s, 2H, CH2), 5.11 (s, 2H, CH2), 5.27 (tt, 1H, J ϭ 6.9
Hz, J ϭ 1.4 Hz, CH), 7.00Ϫ7.29 (m, 3H, aryl), 9.56 (s, 1H, NH).
13C-NMR (CDCl3): δ (ppm) ϭ 13.72 (CH3), 18.04 (CH3), 19.16
(CH2), 25.80 (CH3), 32.78 (CH2), 66.10 (CH2), 72.59 (CH2), 117.46
(C-5), 119.73 (CH), 125.79, 127.72, 135.85, 138.63 (aryl), 138.73
(CϭC), 147.38 (C-2), 151.61 (C-6), 163.07 (C-4). Anal. calcd. for
C19H22Cl2N2O3 (397.30): C, 57.44; H, 5.58; N, 7.05. Found: C,
57.67; H, 5.66; N, 6.90.
6-(3,5-Dichlorobenzyl)-5-ethyl-1-(indan-2-yloxy)methyl-uracil (6i)
1
Yield 277 mg (74%) as a white solid; mp. 166Ϫ167°C. H-NMR
(CDCl3): δ (ppm) ϭ1.04 (t, 3H, J ϭ 7.4 Hz, CH3), 2.40 (q, 2H, J ϭ
7.4 Hz, CH2), 2.89, 3.14 (2 ϫ dd, 4H, J ϭ 3.7, 16.5; 6.1, 16.5 Hz,
1Ј-H, 3Ј-H), 4.09 (s, 2H, CH2), 4.54 (dt, 1H, J ϭ 2.6, 3.7 Hz, 2Ј-
H), 5.18 (s, 2H, CH2), 6.97Ϫ7.28 (m, 7H, Harom.), 9.65 (s, 1H, NH).
13C-NMR (CDCl3): δ (ppm) ϭ 13.75 (CH3), 19.20 (CH2), 32.80
(CH2), 39.45 (CH2), 71.50 (CH2), 79.11 (CH), 117.63 (C-5), 124.67,
125.77, 126.70, 127.71, 135.83, 138.68, 140.29 (Carom.), 147.34 (C-
6), 151.73 (C-2), 163.08 (C-4). Anal. calcd. for C23H22Cl2N2O3
(445.34): C, 62.03; H, 4.98; N, 6.29. Found: C, 62.03; H, 4.94; N,
6.23. 13C-NMR (CDCl3): δ (ppm) ϭ 13.75 (CH3), 19.20 (CH2),
32.80 6.23.
6-(3,5-Dichlorobenzyl)-1-(2-methylallyloxymethyl)-5-ethyluracil (6d)
Yield 190 mg (59%); mp 116Ϫ118°C. 1H-NMR (CDCl3):
δ
(ppm) ϭ 1.08 (t, 3H, J ϭ 7.1 Hz, CH3), 1.72 (s, 3H, CH3), 2.44 (q,
2H, J ϭ 7.2 Hz, CH2), 4.02 (s, 2H, CH2), 4.15 (s, 2H, CH2), 4.90,
4.96 (m, 2H, ϭCH2), 5.15 (s, 2H, CH2), 7.02Ϫ7.30 (m, 3H, aryl),
9.59 (s, 1H, NH). 13C-NMR (CDCl3): δ (ppm) ϭ 13.70 (CH3), 19.18
(CH3), 19.45 (CH2), 32.80 (CH2), 72.77 (CH2), 73.62 (CH2), 112.79
(ϭCH2), 117.56 (C-5), 125.79, 127.78, 135.90, 138.64 (aryl), 140.94
(ϭC), 147.26 (C-2), 151.68 (C-6), 163.08 (C-4). Anal. calcd. for
C18H20Cl2N2O3 (383.28): C, 56.41; H, 5.26; N, 7.31. Found: C,
56.59; H, 5.29; N, 7.30.
Virus and cells
1-((E)-Cinnamyloxymethyl)-6-(3,5-dichlorobenzyl)-5-ethyluracil (6e)
The inhibitory activity against HIV-1 infection was evaluated using
MT-4 cells [16] as target cells and the HIV-1 strain HTLV-IIIB [17]
as infectious virus. The virus was propagated in H9 cells [16] at
37°C, 5% CO2 using RPMI 1640 with 10% heat-inactivated fetal
calf serum (FCS) and antibiotics (growth medium). The culture
supernatant was filtered (0.45 nm), aliquotted, and stored at Ϫ80°C
until use.
Yield 20 mg (5%). 1H-NMR (CDCl3): δ (ppm) ϭ 1.05 (t, 3H, J ϭ
7.5 Hz, CH3), 2.41 (q, 2H, J ϭ 7.4 Hz, CH2), 4.12 (s, 2H, CH2),
4.23 (d, 2H, J ϭ 6.4 Hz, CH2), 5.16 (s, 2H, CH2), 6.15Ϫ6.25 (m,
1H, ϭCHC), 6.56, 6.62 (m, 1H, ϭCHPh), 7.00Ϫ7.21 (m, 3H, aryl),
7.22Ϫ7.42 (m, 5H, Ph). 13C-NMR (CDCl3): δ (ppm) ϭ 13.68
(CH3), 19.20 (CH2), 32.90 (CH2), 70.34 (CH2), 72.55 (CH2), 117.50
(C-5), 124.19, 127.77, 135.87, 138.65 (aryl), 125.77, 126.52, 128.55,
136.18 (Ph), 127.99 (ϭCH), 133.64 (ϭCHPh), 147.23 (C-2), 151.69
(C-6), 162.99 (C-4). HRMS-MALDI: m/z ϭ 467.0900 (M ϩ Naϩ,
C23H22Cl2N2NaO3); requires 467.0903.
Inhibition of HIV-1 replication
Compounds were evaluated for possible antiviral activity against
both strains of HIV-1 using MT-4 cells as target cells. MT-4 cells
were incubated with virus (0.005 MOI) and growth medium con-
taining the test dilutions of compounds for six days in parallel with
virus-infected and uninfected control cultures without compound
added. Expression of HIV in the cultures was indirectly quantified
using the MTT assay [17]. Compounds mediating less than 30%
reduction of HIV expression were considered without biological ac-
tivity. Compounds were tested in parallel for cytotoxic effect in un-
infected MT-4 cultures containing the test dilutions of compounds
as described above. A 30% inhibition of cell growth relative to con-
trol cultures was considered significant. The 50% inhibitory concen-
tration (EC50) and the 50% cytotoxic concentration (CC50) were
determined by interpolation from the plots of percent inhibition
versus concentration of compound. The test for activity against
6-(3,5-Dichlorobenzyl)-1-((E)-2-methyl-3-phenyl-allyloxymethyl)-
5-ethyluracil (6f)
Yield 20 mg (5%). 1H-NMR (CDCl3): δ (ppm) ϭ 1.06 (t, 3H, J ϭ
7.2 Hz, CH3), 1.86 (s, 3H, CH3), 2.44 (q, 2H, J ϭ 7.2 Hz, CH2),
4.15 (s, 2H, CH2), 4.19 (s, 2H, CH2), 5.18 (s, 2H, CH2), 6.49
(ϭCHPh), 7.02Ϫ7.20 (m, 3H, aryl), 7.19Ϫ7.40 (m, 5H, Ph). 13C-
NMR (CDCl3): δ (ppm) ϭ 13.70 (CH3), 15.52 (CH3), 19.20 (CH2),
32.85 (CH2), 72.80 (CH2), 76.03 (CH2), 117.56 (C-5), 125.78 127.79,
135.91, 138.64 (aryl), 125.78, 126.68, 128.12, 134.61 (Ph), 128.86
(ϭCH), 137.00 (ϭCH), 147.26 (C-2), 151.60 (C-6), 162.98 (C-4).
HRMS-MALDI: m/z ϭ 481.1056 (M ϩ Naϩ, C24H24Cl2N2NaO3);
requires 481.1074.
© 2005 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim