Journal of Medicinal Chemistry
ARTICLE
CDCl3: δ 8.04 (d, J = 7.8 Hz, 1H), 7.97 (d, J = 8.8 Hz, 1H), 7.81 (d,
J = 1.9 Hz, 1H), 7.69 (dd, J = 9.8, 2.9 Hz, 1H), 7.42 (d, J = 8.8 Hz, 1H).
1-(4-Methoxyphenyl)methyl-6-bromoindole (12g). 1H NMR: δ
7.73 (s, 1H), 7.51 (br, 2H), 7.17 (d, J = 8.5 Hz, 2H), 7.12 (dd, J =
8.5, 1.2 Hz, 1H), 6.87 (d, J = 8.5 Hz, 2H), 6.49 (d, J = 3.0 Hz, 1H), 5.34
(s, 2H), 3.70 (s, 3H).
3-[[6-(6-Chloro-2-quinolyl)indol-1-yl]methyl]benzoic Acid (14f).
MS calcd for C25H17ClN2O2, 412.8. LC-MS: 415.1 (M + 2)+. 1H
NMR: δ 8.53 (s, 1H), 8.39 (d, J = 8.5 Hz, 1H), 8.24 (d, J = 8.5 Hz, 1H),
8.10 (dd, J = 8.5, 1.8 Hz, 2H), 8.05 (d, J = 9.2 Hz, 1H), 7.83 (dd, J = 6.7,
1.8 Hz, 1H), 7.79 (s, 1H), 7.75 (m, 1H), 7.62 (m, 2H), 7.45 (m, 2H),
6.68 (d, J = 3.0 Hz, 1H), 5.58 (s, 2H).
3-[[6-[1-[(4-Methoxyphenyl)methyl]indol-6-yl]indol-1-yl]methyl]benzoic
Acid (14g). MS calcd for C32H26N2O3, 486. LC-MS: 487.8 (M + H)+.
1H NMR: δ 7.83 (br, 2H), 7.69 (d, J = 8.5 Hz, 2H), 7.60 (t, J = 8.5 Hz,
2H), 7.55 (t, J = 3.0 Hz, 2H), 7.46 (br, 2H), 7.35 (m, 2H), 7.20 (d, J = 8.5
Hz, 2H), 6.84 (d, J = 8.5 Hz, 2H), 6.51 (d, J = 3.0 Hz, 1H), 6.45 (d, J =
3.0 Hz, 1H), 5.60 (s, 2H), 5.39 (s, 2H), 3.66 (s, 3H).
3-[[6-[1-[(4-Fluoro)methyl]indol-6-yl]indol-1-yl]methyl]benzoic Acid
(14h).MS calcd for C31H23FN2O2,474.2.LC-MS:473(Mꢀ H)ꢀ.1HNMR:
δ 7.82 (br, 2H), 7.69 (d, J = 4.8 Hz, 2H), 7.60 (t, J = 8.8 Hz, 2H), 7.54 (d, J =
2.9 Hz, 1H), 7.51 (d, J= 2.9 Hz, 1H), 7.45 (m, 2H), 7.35 (m, 2H), 7.27 (dd, J=
6.8, 5.8 Hz, 2H), 7.12 (t, J = 8.8 Hz, 2H), 6.51 (d, J = 2.9 Hz, 1H), 6.48
(d, J = 2.9 Hz, 1H), 5.59 (s, 2H), 5.48 (s, 2H).
3-[[5-(6-Chloro-2-quinolyl)indol-1-yl]methyl]benzoic Acid (14i).
MS calcd for C25H17ClN2O2, 413. LC-MS: 415.3 (M + 2)+. 1H
NMR: δ 8.42 (s, 1H), 8.40 (d, J = 9.2 Hz, 1H), 8.27 (d, J = 8.5 Hz,
1H), 8.10 (d, J = 2.4 Hz, 1H), 8.04 (d, J = 8.5 Hz, 2H), 7.81 (br, 2H), 7.72
(m, 2H), 7.67 (d, J = 3.0 Hz, 1H), 7.48 (m, 2H), 6.59 (d, J = 2.4 Hz, 1H),
5.67 (s, 2H).
6-(3-Hydroxyphenyl)-1H-indole (13b). MS calcd for C14H11NO,
1
209. LC-MS: 210.5 (M + H)+. H NMR: δ 9.46 (s, <1H, exch), 7.58
(m, 2H), 7.38 (t, J = 2.9 Hz, 1H), 7.23 (m, 2H), 7.07 (d, J = 7.8 Hz, 1H),
7.04 (d, J = 1.9 Hz, 1H), 6.71 (dd, J = 7.8, 1.9 Hz, 1H), 6.44 (s, 1H).
6-(3-Methoxyphenyl)-1H-indole (13c). MS calcd for C15H13NO,
223. LC-MS: 224.6 (M + H)+. 1H NMR: δ 11.16 (s, <1H, exch), 7.63
(s, 1H), 7.59 (d, J = 7.9 Hz, 1H), 7.37 (t, J = 3.0 Hz, 1H), 7.34 (d, J = 7.9
Hz, 1H), 7.30 (dd, J = 7.9, 1.8 Hz, 1H), 7.23 (d, J = 7.9 Hz, 1H), 7.17 (br,
1H), 6.89 (dd, J = 8.5, 1.8 Hz, 1H), 6.44 (s, 1H), 3.82 (s, 3H).
6-[3-[(4-Methoxyphenyl)methoxy]phenyl]-1H-indole (13e). 1H NMR
(400 MHz): δ 7.49 (s, 1H), 7.38 (s, 1H), 7.34 (s, 1H), 7.33 (d, J = 1.9 Hz,
1H), 7.28 (t, J = 7.8 Hz, 1H), 7.17 (d, J = 8.8 Hz, 1H), 7.11 (d, J = 2.9 Hz,
2H), 7.07 (d, J = 5.9 Hz, 1H), 6.96 (d, J = 9.8 Hz, 1H), 6.90 (d, J = 8.8 Hz,
2H), 6.87 (dd, J = 8.8, 1.9 Hz, 1H), 5.04 (s, 2H), 3.74 (s, 3H).
6-(6-Chloro-2-quinolyl) Indole (13f). MS calcd for C11H17ClN2, 278.
LC-MS: 280.8 (M + 2)+. 1H NMR in CDCl3: δ 8.33 (s, 1H), 8.12 (t, J =
8.8 Hz, 2H), 7.99 (d, J = 8.8 Hz, 1H), 7.90 (dd, J = 7.8, 1.9 Hz, 1H), 7.81
(d, J = 1.9 Hz, 1H), 7.78 (d, J = 7.8 Hz, 1H), 7.65 (dd, J = 8.8, 1.9 Hz,
1H), 7.32 (t, J = 2.9 Hz, 1H), 6.62 (s, 1H).
6-[1-[(4-Methoxyphenyl)methyl]indol-6-yl]-indole (13g). MS calcd
for C24H20N2O, 352. LC-MS: 335.8 (M + H ꢀ H2O)+. 1H NMR: δ 7.72
(s, 1H), 7.68 (m, 1H),7.62 (s, 1H), 7.61 (d, J = 3.6 Hz, 1H), 7.59 (d, J =
4.3 Hz, 1H), 7.48 (d, J = 3.0 Hz, 1H), 7.34 (s, 1H), 7.31 (d, J = 7.3 Hz,
1H), 7.21 (d, J = 8.5 Hz, 2H), 6.87 (d, J = 8.5 Hz, 2H), 6.47 (d, J = 3.0 Hz,
1H), 6.43 (s, 1H), 5.42 (s, 2H), 3.68 (s, 3H).
5-(6-Chloro-2-quinolyl) Indole (13i). MS calcd for C11H17ClN2, 278.
LC-MS: 279.1 (M + H)+. 1H NMR in CDCl3: δ 8.45 (s, 1H), 8.11 (m,
3H), 7.99 (d, J = 8.8 Hz, 1H), 7.91 (dd, J = 11.0, 7.8 Hz, 1H), 7.80 (d, J =
1.9 Hz, 1H), 7.64 (dd, J = 8.8, 1.9 Hz, 1H), 7.54 (d, J = 8.8 Hz, 1H), 7.19
(t, J = 2.9 Hz, 1H), 6.69 (s, 1H).
3-((3-(1-(3-Carboxybenzyl)-1H-indol-6-yl)phenoxy)methyl)benzoic
Acid (14b). MS calcd for C30H23NO5, 477. LC-MS: 478.1 (M + H)+. 1H
NMR: δ 8.07 (s, 1H), 7.91 (d, J = 7.8 Hz, 1H), 7.79 (m, 3H), 7.73 (d, J =
7.8 Hz, 1H), 7.62 (d, J = 7.8 Hz, 1H), 7.56 (d, J = 3.9 Hz, 1H), 7.53 (d, J =
7.8 Hz, 1H), 7.44 (m, 2H), 7.33 (m, 2H), 7.30 (d, J = 1.9 Hz, 1H), 7.25
(d, J = 7.8 Hz, 1H), 6.97 (dd, J = 8.8,1.9 Hz, 1H,), 6.53 (d, J = 2.9 Hz,
1H), 5.60 (s, 2H), 5.26 (s, 2H).
3-((6-(3-Methoxyphenyl)-1H-indol-1-yl)methyl)benzoic Acid
(Compound 14c). MS calcd for C23H19NO3, 357. LC-MS: 358.7
(M + H)+. 1H NMR: δ 7.82 (s, 1H), 7.81 (s, 1H),7.77 (s, 1H), 7.62 (d, J =
8.5 Hz, 1H), 7.57 (d, J = 2.4 Hz, 1H), 7.46 (m, 2H), 7.34 (m, 2H), 7.23
(d, J = 7.3 Hz, 1H), 7.18 (s, 1H), 6.88 (dd, J = 7.9, 1.9 Hz, 1H), 6.53 (d,
J = 2.9 Hz, 1H), 5.60 (s, 2H), 3.82 (s, 3H).
3-((6-(3-Hydroxyphenyl)-1H-indol-1-yl)methyl)benzoic Acid (14d).
MS calcd for C22H17NO3, 343. LC-MS: 344.1 (M + H)+. 1H NMR: δ
8.07 (s, 1H), 7.90 (d, J = 7.8 Hz, 1H), 7.74 (d, J = 6.8 Hz, 1H), 7.63 (s,
1H), 7.59 (d, J = 8.8 Hz 1H), 7.54 (t, J = 7.8 Hz, 1H), 7.36 (m, 2H), 7.29
(br, 2H), 7.25 (d, J = 7.8 Hz, 1H), 6.97 (d, J = 7.8 Hz, 1H), 6.44 (s, 1H),
5.28 (s, 2H).
3-((3-((6-(3-(4-Methoxybenzyloxy)phenyl)-1H-indol-1-yl)methyl)
benzoyloxy)methyl) benzoic Acid (14e). MS calcd for C39H31NO6,
597. LC-MS: 598.1 (M + H)+. 1H NMR: δ 8.02 (s, 1H), 7.89 (d, J = 7.8
Hz, 1H), 7.78 (d, J = 6.8 Hz, 1H), 7.75 (s, 1H), 7.68 (d, J = 7.8 Hz, 1H),
7.57 (br, 2H), 7.50 (t, J = 7.8 Hz, 1H), 7.37 (m, 2H), 7.31 (s, 1H), 7.07
(d, J = 8.8 Hz, 1H), 6.92 (m, 2H), 6.84 (d, J = 2.9 Hz, 1H), 6.69 (m, 5H),
6.54 (d, J = 2.9 Hz, 1H), 5.45 (s, 2H), 5.17 (s, 2H), 3.71 (s, 2H),
3.66 (s, 3H).
3-(1H-Indol-6-yl)benzoic Acid (15). MS calcd for C15H11NO2, 237.
1
LC-MS: 220.1 (M + H ꢀ H2O)+. H NMR: δ 11.21 (s, 1H), 8.21
(s, 1H), 7.93 (d, J = 7.3 Hz, 1H), 7.89 (d, J = 7.3 Hz, 1H), 7.67 (s, 1H), 7.64
(d, J = 7.3 Hz, 1H), 7.58 (t, J = 7.9 Hz, 1H), 7.41 (br, 1H), 7.33 (d, J = 7.9
Hz, 1H), 6.47 (s, 1H).
Binding Affinity Assay. Inhibition constants KI for binding in the
hydrophobic pocket were determined using a fluorescence intensity
assay as previously described.23,24 Briefly, a metallopeptide receptor
structure FeII(env2.0)3 was used to mimic the hydrophobic pocket in the
gp41 coiled coil. Env2.0 contains 17 hydrophobic pocket residues
flanked by five residues on either side. Mixing FeII(env2.0)3 with a
fluorescein labeled pocket-binding C-peptide C18-FL caused quenching
of fluorescence, which could be reversed in the presence of a competitive
inhibitor. Typically, 7 μM binding sites (three per receptor trimer) and
7.5 nM C18-FL were used in the assay.
CCF Assay. CCF was measured following a published procedure25
and using cell lines obtained through the NIH AIDS Research and
Reference Reagent Program, Division of AIDS, NIAID, NIH. TZM-bl
cells (#8129, contributed by J. C. Kappes, X. Wu, and Tranzyme Inc.)
expressing CD4, CCR5, and CXCR438 and containing an integrated
reporter gene for firefly luciferase under control of HIV-1 LTR39 were
used as target cells. They were grown overnight in 96 well plates in
Dulbecco's modified Eagle medium (DMEM) supplemented with 10%
fetal bovine serum (FBS), using 25000 cells per well. The following day,
the medium was exchanged with reduced serum medium (Gibco), and
1 μL of compound in DMSO was added to each well, using 6ꢀ10 serial
dilutions to obtain doseꢀresponse curves. HL2/3 effector cells (#1294,
contributed by B. K. Felber and G. N. Pavlakis), which produce HXB2
Env, Tat, and Rev40 were added, using 50000 cells per well, to a total well
volume of 100 μL. After 6 h, luciferase expression was measured using
Luciferase Assay Reagent (Promega). Controls containing 1 μL of
DMSO with and without HL2/3 cells were measured for each com-
pound, and experiments were performed in duplicate.
Viral Replication Assay. Inhibition of HIV-1 replication was
determined in CCR5- and CXCR4-tropic MAGI antiviral assays as
previously described.26,27 HIV-1 isolates and cells were obtained from
the NIH AIDS Research and Reference Reagent Program, Division of
AIDS, NIAID, NIH, as follows: HIV-1 Ba-L from Suzanne Gartner,
Mikulas Popovic, and Robert Gallo.26,41 HIV-1 IIIB from Robert C.
Gallo26,42 MAGI-CCR5 cells from Dr. Julie Overbaugh.43,44 Briefly,
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dx.doi.org/10.1021/jm200791z |J. Med. Chem. 2011, 54, 7220–7231