Synthesis and SAR of indazole-pyridine based protein kinase B/Akt inhibitors

Article abstract of DOI:10.1016/j.bmc.2006.06.047

A series of heteroaryl-pyridine containing inhibitors of Akt are reported. The synthesis and structure-activity relationships are discussed, leading to the discovery of a indazole-pyridine analogue (K_i = 0.16 nM). These compounds bind in the ATP binding site, are potent, ATP competitive, and reversible inhibitors of Akt activity. No selectivity amongst the Akt isoforms is observed for this analogue, but there is good selectivity against an panel of other kinases. It is least selective for other members of the AGC family of kinases but is nonetheless 40-fold selective for Akt over PKA. The compound shows cellular activity and significantly slows tumor growth in vivo.

Full text of DOI:10.1016/j.bmc.2006.06.047

Bioorganic & Medicinal Chemistry 14 (2006) 6832–6846
Synthesis and SAR of indazole-pyridine based protein
kinase B/Akt inhibitors
Keith W. Woods,a,* John P. Fischer,a Akiyo Claiborne,a Tongmei Li,a Sheela A. Thomas,a
Gui-Dong Zhu,a Robert B. Diebold,a Xuesong Liu,a Yan Shi,a Vered Klinghofer,a
Edward K. Han,a Ran Guan,a Shayna R. Magnone,a Eric F. Johnson,a Jennifer J. Bouska,a
Amanda M. Olson,a Ron de Jong,b Tilman Oltersdorf,b Yan Luo,a
Saul H. Rosenberg,a Vincent L. Girandaa and Qun Lia
aCancer Research, Department R47S, AP10, Abbott Laboratories, 100 Abbott Park Rd., Abbott Park, IL 60064, USA
bIdun Pharmaceuticals, 9380 Judicial Dr., San Diego, CA 92121, USA
Received 1 June 2006; revised 12 June 2006; accepted 19 June 2006
Available online 14 July 2006
Abstract—A series of heteroaryl-pyridine containing inhibitors of Akt are reported. The synthesis and structure–activity relation-
ships are discussed, leading to the discovery of a indazole-pyridine analogue (Ki = 0.16 nM). These compounds bind in the ATP
binding site, are potent, ATP competitive, and reversible inhibitors of Akt activity. No selectivity amongst the Akt isoforms is
observed for this analogue, but there is good selectivity against an panel of other kinases. It is least selective for other members
of the AGC family of kinases but is nonetheless 40-fold selective for Akt over PKA. The compound shows cellular activity and
significantly slows tumor growth in vivo.
ꢀ 2006 Elsevier Ltd. All rights reserved.
1. Introduction
All three Akt isoforms are either overexpressed or acti-
vated in a variety of human tumors including lung,
breast, prostate, ovarian, gastric, and pancreatic carci-
nomas.12–14 Increased levels of Akt expression also cor-
relate with disease progression. The central role of Akt
in growth and survival pathways provides the rationale
for inhibiting Akt in the treatment of cancer. Several
Akt inhibitors have been reported.15–21
Akt1 (also called protein kinase B, PKB)1–3 is a serine/
threonine protein kinase that exhibits elevated activity
in a large proportion of human malignancies.4,5 Akt1
was discovered as the human homologue of the trans-
forming gene in the Akt-8 oncogenic virus which was
isolated from a spontaneous thymoma in the AKR
mouse.6,7 Two additional Akt isoforms, Akt2 and
Akt3, have been identified. The three Akts are approxi-
mately 85% homologous in protein sequence.8–10 Se-
quence homology in the ATP binding site is 100%
except for one non-crucial amino acid in Akt3. Akt is
a member of the AGC family of kinases and has a high
degree of homology with PKA and PKC.11
Herein, we report the development of a series of Akt
kinase inhibitors. These compounds are potent, ATP
competitive, reversible inhibitors of Akt. Our efforts
have resulted in the discovery of the indazole-pyridine
compound 4. The compound is a potent (Akt1
Ki = 0.16 nM) and selective Akt inhibitor (greater than
20-fold selective for Akt over more than 35 other
kinases), and causes significant delay in the growth of
tumors in mouse xenograft models.22
Akt is activated by phosphorylation in response to a
number of mitogenic stimuli. Fully activated Akt is
phosphorylated at two sites: Thr 308 and Ser 473.
2. Chemical synthesis
Keywords: Akt; Protein kinase B; Serine/threonine kinase; Indazole.
*
Compound 1 (Fig. 1) was identified as a lead compound
from
Corresponding author. Tel.: +1 847 938 2808; fax: +1 847 935
5165; e-mail: keith.w.woods@abbott.com
a
high-throughput screening assay (Akt1
0968-0896/$ - see front matter ꢀ 2006 Elsevier Ltd. All rights reserved.
doi:10.1016/j.bmc.2006.06.047

K. W. Woods et al. / Bioorg. Med. Chem. 14 (2006) 6832–6846
6833
N
Cl
N
NH2
NH2
O
O
N
N
1
2
NH
N
N
H3C
N
NH2
NH2
O
O
N
N
H
NH
NH
3
4
Figure 1. Hit to lead progression.
Ki = 5 lM). The chlorine was removed for simplicity
and investigation of the ether-linked side-chain require-
ments was undertaken. These efforts have been reported
previously.16 That research led to the discovery of
compound 2 with the indole containing side chain.
Compound 2 (Akt1 Ki = 14 nM) represents an increase
in the Akt1 activity of greater than 350-fold when
compared to the initial screening hit.
The preparation of compound 3 required the synthesis
of 6-bromoisoquinoline. The 6-bromoisoquinoline was
initially prepared according to the procedure reported
by Hendrickson and Rodriguez.23 This method suffers
from long reaction times (>4 days) and modest yields.
We have thus found the synthesis described by Miller
and Frincke to be preferable.24
The SAR studies carried out to investigate the impor-
tance of the isoquinoline moiety are summarized in
Table 1. The aromatic starting materials required for
the preparation of compounds 37, 38, 43, 44, 46, and
47 are commercially available.
The details of restricting rotation about the olefin link-
age of 2, resulting in compound 3 with significantly
improved Akt1 potency (Akt1 Ki = 0.99 nM), were
discussed in an earlier communication from our labora-
tories.17 Extensive SAR studies to investigate replace-
ments for the isoquinoline ring in 3 have yielded the
highly active indazole containing Akt inhibitor 4 (Akt1
Ki = 0.16 nM). These studies are detailed here.
Example 39 requires the preparation of 6-bromocinno-
line (16). The synthesis is outlined in Scheme 2.
Acetylation of 2-aminoacetophenone (10) followed by
bromination with bromine in acetic acid gives 12.
De-acetylation and diazotization result in cyclization
to the cinnolinone 13.25 Upon treatment with phos-
phorus oxychloride, the 4-chlorocinnolinone 14 is
formed. Displacement of the chlorine with hydrazine
gives compound 15. Heating the hydrazino intermedi-
ate in the presence of aqueous CuSO4 leads to removal
of the hydrazine and isolation of the desired 6-bromo-
cinnoline 16.
The general synthesis of the compounds is outlined in
Scheme 1. The side-chain ether linkage is made via a
Mitsunobu reaction between N-Boc-tryptophanol (6)
and 3-bromo-5-hydroxypryidine (5). The aryl or hetero-
aryl moieties are introduced using the Stille reaction
rather than alternate methods such as the Suzuki reac-
tion since, in general, the Stille reaction proved to be
more effective. Finally, removal of the Boc protecting
group yielded the desired Akt inhibitors. Deprotection
with HCl was usually unsuccessful. It appears that
protonation, and subsequent precipitation of the
compounds, occurs faster than deprotection. TFA in
CH2Cl2 leads to the desired deprotected amines.
The three isoquinolines (40–42) with substitutions in the
1-position derive from 6-bromo-1-hydroxyisoquinoline
(17) which was prepared according to a literature proce-
dure (Scheme 3).26 Treatment of 17 with phosphorus
N
NHBoc
N
HO
i
ii
+
NHBoc
Br
O
Br
OH
N
H
5
6
7
N
H
N
N
R
R
NHBoc
NH2
Br
Me3Sn
O
O
iii, iv
+
N
N
N
N
H
H
N
H
N
H
8
9
Scheme 1. Reagents and conditions: (i) DEAD, PPh3, THF, 0 ꢁC to rt, overnight, 89%; (ii) Me3SnSnMe3, Pd(PPh3)4, 75 ꢁC, 1.5 days, 34%;
(iii) Pd2(dba)3, P(o-tol)3, TEA, 110 ꢁC, 4 h, 49%; (iv) TFA, CH2Cl2, rt, 3 h, 81%.

6834
K. W. Woods et al. / Bioorg. Med. Chem. 14 (2006) 6832–6846
Table 1. Structures and in vitro Akt1 kinase bindinga
Table 1 (continued)
N
Compound
Ar
Akt1 Ki (nM)
NH2
Ar
O
52
53
2.6
N
NH
N
H
Compound
Ar
Akt1 Ki (nM)
3.9
37
1117
N
N
H
3b
0.99
312
N
54
55
174
N
N
H
38
39
40c
N
S
S
1.3
N
N
215
N
N
N
H
N
N
N
1022
56
9.8
OH
N
H
41c
188
HN
HN
NH2
57
58
1.2
9.8
N
N
N
H
42c
43
473
331
N
N
Cl
N
N
N
H
H2N
N
44b
45b
46
245
59
60
2.9
18
N
H
H
N
N
1659
4022
1503
0.99
N
O
N
O
N
H
HO
N
61
62
18
47b
48
N
NC
N
N
H
O
HO
N
1848
H
N
N
H3C
N
H
H
N
4
(S-isomer)
(R-isomer)
0.16
28
63
64
237
N
N
H
H
H3C
N
N
1232
49
N
N
H
H2N
N
N
H3C
65
92
N
N
50
51
685
H
N
a Values were measured against Akt1 with ATP concentrations of
10 lM.22,36
H3C
b Compound was reported in Ref. 17. The data are shown here for the
ease of the reader.
1.15
N
N
c Compound was reported in Ref. 18. The data are shown here for the
ease of the reader.
H

K. W. Woods et al. / Bioorg. Med. Chem. 14 (2006) 6832–6846
6835
O
O
O
O
Br
iii
ii
Br
i
N
HN
HN
N
H
H2N
O
O
10
11
12
13
Cl
N
NHNH2
iv
v
Br
Br
vi
Br
N
N
N
N
N
14
15
16
Scheme 2. Reagents and conditions: (i) AcCl, TEA, CH2Cl2, rt, 3 h, 100%; (ii) Br2, HOAc, 75 min, 89%; (iii) a—HCl (aq), THF, reflux, 1 h; b—6 N
HCl, NaNO2, 0 ꢁC, 2 h then overnight at rt then reflux 6 h, 54%; (iv) POCl3, 100 ꢁC 2 h, 43%; (v) H2NNH2ÆH2O, EtOH, rt, 3 days, 100%; (vi) CuSO4,
H2O, reflux, 2 h, 24%.
Br
Br
2-fluorobenzaldehyde (22) with hydrazine (Scheme 5). A
series of compounds having substitutions at the 3-posi-
tion of the indazole are shown in Table 1.
i
ii
N
N
OH
17
Cl
N
18
Carbon substitutions in the 3-position of the indazoles
(4, 49, 51, 52, 53, 54, 55, 56, and 58) are prepared by an-
ion addition (Grignard reagents or lithiated species) to
5-bromo-2-fluorobenzaldehyde (22) (Scheme 5). The
resulting alcohol is oxidized with manganese dioxide
to give the corresponding ketone 25. The indazole 26
is then formed by refluxing the ketone in hydrazine.
Br
Br
iii
N
NH2
N(Boc)2
19
20
Scheme 3. Reagents and conditions: (i) POCl3, 100 ꢁC, 4 h, 62%; (ii)
acetamide, K2CO3, 180 ꢁC, 5 h, 65%; (iii) Boc2O, DMAP, TEA,
CH3CN, rt, 2 h, 71%.
The pyrrole-substituted indazole required for the syn-
thesis of 57 is prepared as outlined in Scheme 6. The acid
chloride 29 is obtained by treatment of 5-bromo-2-flu-
orobenzoic acid (28) with thionyl chloride. Friedel–
Crafts reaction of 29 with pyrrole gives the requisite
ketone 30 which is converted to the indazole 31 under
the usual treatment with hydrazine.
oxychloride yields 6-bromo-1-chloroisoquinoline (18).
The chloride is displaced with acetamide which under-
goes hydrolysis to give 1-amino-6-bromoisoquinoline
(19). The amino group is bis protected with di-tert-butyl
dicarbonate to give 20 prior to the Stille coupling
reaction.
Coupling of the 5- or 6-bromo-3-aminoindazole with the
stannylpyridine intermediate 8 using the usual Stille con-
ditions fails to give the desired product (59 or 64). These
compounds are therefore obtained by doing the Stille
reaction prior to formation of the indazole. Reaction
of stannane 8 with 5-bromo-2-fluorobenzonitrile (32)
followed by formation of the indazole ring system gives
Synthesis of the amide linkage (45) begins with carbon-
ylation of the bromopyridine intermediate 7 to give car-
boxylic acid 21 (Scheme 4). Carbodiimide activation of
the carboxyl group followed by addition of 4-amino-
pyridine and removal of the Boc protecting group yields
45.
5-Bromoindazole (23) which is used for the preparation
of compound 48 is prepared by the reaction of 5-bromo-
O
O
F
R
Br
Br
Br
Br
i,ii
H
iii
N
R
N
N
i
N
F
NHBoc
HO
NHBoc
H
O
Br
O
22
iii
25
26
O
vi
CH3
N
H
7
21
N
H
R
H3C
N
Br
Br
iv, v
N
N
N
H3C
N
N
N
H
H
N
ii, iii
N
NH2
H
O
23
24
27
(R=CH3)
N
O
N
Scheme 5. Reagents and conditions: (i) RMgBr, Et2O, 0 ꢁC, 1 h, 85–
99%; (ii) MnO2, p-dioxane, reflux, 4 h, 78–85%; (iii) H2NNH2ÆH2O,
reflux, 9 h, 60–95%; (iv) fuming HNO3, Ac2O, HOAc, ꢀ5 ꢁC, 45 min,
94%; (v) Me2NH, THF, reflux, overnight, 45%; (vi) NaH, DMF, CH3I,
rt, 2 h, 67%.
H
45
Scheme 4. Reagents and conditions: (i) PdCl2Ædppf, CO, THF/H2O,
100 ꢁC, 19 h, 76%; (ii) 4-aminopyridine, EDCI, HOBt, DMF, rt,
overnight, 18%; (iii) HCl/dioxane, CH2Cl2, 2 h, rt, 31%.

6836
K. W. Woods et al. / Bioorg. Med. Chem. 14 (2006) 6832–6846
O
F
Br
Br
N
N
H2N
H2N
Br
N
O
F
ii
i
Br
N
N
i
HO
Br
+
N
H
Cl
N
Boc
H
34
35
36
28
29
Scheme 8. Reagents and conditions: (i) NaNO2, H2SO4, 30 min, 72%;
(ii) a—phosgene, THF, ꢀ20 ꢁC, 1 h, then rt, 2 h; b—t-BuOH, THF,
ꢀ20 ꢁC to rt, overnight, 76%.
O
H
N
HN
Br
iii
ii
Br
N
F
efforts that resulted in the discovery of the indole con-
taining side chain (2, Akt1 Ki = 14 nM) and the work
leading to the isoquinoline containing compound (3,
Akt1 Ki = 0.99 nM) have been reported previously.16,17
We now report the SAR studies leading to the discovery
of the indazole moiety as a replacement for the isoquin-
N
H
30
31
Scheme 6. Reagents and conditions: (i) SOCl2, reflux, 2 h; (ii) AlCl3,
1,2-dichloroethane, 0 ꢁC to rt, overnight, 31%; (iii) ) H2NNH2ÆH2O,
reflux, 9 h, 95%.
oline. The 3-methylindazole analogue
4
(Akt1
compound 59 (Scheme 7). The 3-aminoindazole at-
tached to the pyridine ring at the indazole 6-position
(64) can be prepared in the same manner as described
for compound 59 by starting with 4-bromo-2-
fluorobenzonitrile.
Ki = 0.16 nM) is the most potent Akt inhibitor obtained
from this work. It shows good selectivity and has shown
in vivo efficacy in several mouse tumor models.22
The ring nitrogen in both the pyridine ring of 2 and the
isoquinoline ring of 3 appears to be well oriented to
interact favorably in the hinge binding region of the
ATP binding site. This hinge binding interaction is a
common motif observed for most ATP competitive
kinase inhibitors.
The 5-bromo-3-dimethylaminoindazole required for the
synthesis of 60 and 5-bromo-3-morpholinoindazole used
in the preparation of 61 are prepared according to the
procedure reported by Wrzeciono et al.27 Compound
26 (R = CH3) is treated with iodomethane in the pres-
ence of base to give the 5-bromo-1,3-dimethylindazole
analogue 27 (Scheme 5). The 6-substituted compounds
(63 and 64) are prepared as described for compounds
48 and 59, respectively, replacing the 5-bromobenzene
starting materials with the corresponding 4-bromoben-
zene reagents.
The details of our efforts investigating replacements of
the olefin linkage have been reported earlier.17 In sum-
mary, we found that if the olefin link in 2 is removed,
the activity of the compounds decreases (44 Akt1
Ki = 245 nM). The pyridine is too far away from the
hinge binding region to be optimal if the other favorable
molecular interactions are maintained in the ATP bind-
ing site. Attempts to interact with the hinge binding re-
gion with substituted phenyl rings were unsuccessful (46,
Akt1 Ki = 4022 nM; 47, Akt1 Ki = 1503 nM). Exchang-
ing the olefin with an amide linkage, likewise, leads to
a loss of Akt1 activity (45, Akt1 Ki = 1659 nM).
The 5-bromobenztriazole (35) necessary for the synthe-
sis of 65 is obtained in two steps from 4-bromo-1,2-di-
aminobenzene (34) (Scheme 8). Diazotization of the
diamine 34 yields the benztriazole 35. The ring nitrogen
is protected with a Boc group (36) prior to Stille
coupling.28
Restricting rotation about the olefin linkage of 2, how-
ever, results in compound 3 with significantly improved
Akt1 potency (Akt1 Ki = 0.99 nM).17 This represents an
improvement in activity of 14-fold versus compound 2
and an increase in activity of greater than 5000-fold over
the initial hit (1).
3. Results and discussion
Compound 1 (Fig. 1) was identified as a hit from a high-
throughput screening assay (Akt1 Ki = 5 lM). The SAR
N
N
NC
F
Br
i
NC
F
NHBoc
NHBoc
O
+
Me3Sn
O
32
8
33
N
H
N
H
N
H2N
ii, iii
NH2
O
N
N
H
N
59
H
Scheme 7. Reagents and conditions: (i) Pd2(dba)3, P(o-tol)3, TEA, 110 ꢁC, 4 h, 49%; (ii) H2NNH2ÆH2O, reflux, 5 h, 84%; (iii) TFA, CH2Cl2, rt 3 h,
81%.

K. W. Woods et al. / Bioorg. Med. Chem. 14 (2006) 6832–6846
Table 2. Selectivity of Akt inhibitors for selected kinasesa
Family Kinase Ki, nM (selectivity)
Compound 48 Compound 4b
6837
Efforts were, therefore, undertaken to investigate the
importance of the isoquinoline nitrogen and the require-
ments with respect to its regiochemical orientation. As
would be predicted, the presence of the nitrogen atom
is essential for Akt inhibitory activity. Replacing the iso-
quinoline with a naphthyl (37) results in a large decrease
in potency (Akt1 Ki = 1117 nM). Removal of the iso-
quinoline nitrogen eliminates the ability to hydrogen
bond to the protein backbone. Likewise, the position
of the nitrogen atom in the hinge binding interaction
is important. The quinoline analogue (38) shows de-
creased Akt activity (Akt1 Ki = 312 nM) as does the iso-
quinoline analogue connected to the pyridine through
the isoquinoline 5-position (43, Akt1 Ki = 331 nM).
Not only are the presence and position of the nitrogen
important, there also appears to be a sensitivity to the
basicity of the nitrogen in heteroaromatic ring. The less
basic cinnoline rings system (39) maintains a nitrogen in
the same spatial orientation as the isoquinoline but
shows decreased Akt1 activity (Akt1 Ki = 215 nM) sim-
ilar to either the quinoline (38, Akt1 Ki = 312 nM) or the
5-substituted isoquinoline (43, Akt1 Ki = 331 nM)
analogues.
Compound 3
AGC
Akt1
PKA
0.99
2.1 (2.1)
0.99
19 (19)
0.16
6.3 (40)
33 (200)
24 (150)
82 (512)
PKCd 225 (225)
PKCc 314 (314)
222 (222)
182 (182)
777 (777)
SGK
270 (270)
CMGC CDK2 82 (82)
CDC2 85 (85)
ERK2 524 (524)
42 (42)
257 (257)
633 (633)
24 (150)
127 (794)
340 (2100)
2400 (15,000)
CK2
CAMK MAPK 13,800 (13,800) 8160 (8160)
TK SRC 2180 (2180) 5000 (5000)
13,600 (13,600) 5270 (5270)
3300 (21,000)
2600 (16,000)
a Ki value is shown (nM). The fold selectivity is shown in parentheses.
b Some of the selectivity data for compound 4 have been reported in
Ref. 22. The data are re-reported here for the ease of the reader.
A number of compounds were prepared with substitu-
tions in the 3-position of the indazole. A variety of
groups were tolerated on the indazole. The thiazole 56
and imidazole 58 analogues had Akt1 Ki values of
9.8 nM. The phenyl analogue 53 had intermediate activ-
ity (Akt1 Ki = 3.9 nM). The thiophene 55 (Akt1
Ki = 1.3 nM) and pyrrole 57 (Akt1 Ki = 1.2 nM) ana-
logues both had Akt1 activity less than 2 nM. Methyl
4 (Akt1 Ki = 0.16 nM), ethyl 51 (Akt1 Ki = 1.1 nM),
and cyclopropyl 52 (Akt1 Ki = 2.6 nM) analogues all
had Akt1 Ki values less than 3 nM. Introduction of a
methylene linkage between the indazole and the phenyl
ring (54) resulted in decreased activity (Akt1
Ki = 174 nM).
Pharmacokinetic examination of the isoquinoline com-
pound 3 suggested metabolic liabilities with the mole-
cule.29 The 1-position of the isoquinoline appears to be
prone to oxidative metabolism. The 1-hydroxy com-
pound was prepared (40) and was found to have little
activity (Akt1 Ki = 1022 nM). Attempts to prevent oxi-
dative metabolism by blocking the 1-position likewise
resulted in compounds with diminished Akt1 activity
(41, Akt1 Ki = 188 nM and 42, Akt1 Ki = 473 nM). De-
tails of the SAR studies investigating the isoquinoline
moiety are reported elsewhere.18
An unsubstituted amino group 59 in the 3-position was
tolerated (Akt1 Ki = 2.9 nM) but the activity decreased
to some extent with substituted amines. Dimethylamino
60 and morpholino 61 both showed Akt1 Ki values of
18 nM. Introduction of a carboxylic acid (62), on the
other hand, was unfavorable. There was a dramatic
decrease in the Akt1 inhibitory activity (Akt1
Ki = 1848 nM).
We desired to find an alternative to the isoquinoline
ring. Replacement of the isoquinoline with indazole
(48) was found to give a compound with equal potency
to 3 (Akt1 Ki = 0.99 nM) while eliminating the oxida-
tive liabilities of the isoquinoline. In addition, it was
found that the indazole compound 48 exhibited an im-
proved selectivity profile versus other kinases (see
Table 2).
The indazole interacts with the hinge binding region of
the enzyme through a hydrogen bond donor–acceptor
relationship.16,17,22 The free N–H of the indazole is
essential for this interaction. Methylation of the N-1
position (50) disrupts that binding possibility and results
in a dramatic decrease in activity (Akt1 Ki = 685 nM).
Many of the compounds were examined for their selec-
tivity for Akt over other kinases. Akt1, Akt2, and
Akt3 are approximately 85% homologous in protein
sequence, and as a result, little selectivity is observed
amongst the Akt isoforms. Selected compounds were
tested against a panel of no fewer than 15 kinases.
(See Table 2 for some representative examples.) Akt
is a Ser/Thr kinase and, as would be expected, the
compounds are highly selective for Akt when com-
pared to tyrosine kinases (>1000-fold). The com-
pounds are least selective for the closely related
AGC family of kinases (e.g., PKA and PKC). Com-
pound 3 was only 2-fold selective for Akt over PKA,
whereas 48 showed a selectivity of nearly 20-fold.
The Akt activity, the better selectivity profile, and
the improved metabolic profile of the indazole com-
pound 48 prompted us to further investigate the
SAR about the indazole ring.
A stereochemical preference is observed for the ether-
linked side chain. Both enantiomers of the tryptophanol
side chain were prepared. The S-stereoisomer of the
ether side-chain 4 (Akt1 Ki = 0.16 nM) is preferred over
the R-enantiomer 49 (Akt1 Ki = 28 nM).30
The attachment in the 5-position of the indazole to the
pyridine (48) is optimal for Akt activity. Attachment
in the 6-position of the indazole (63) results in a loss
of Akt activity (Akt1 Ki = 237 nM). The orientation of
the hydrogen bond donor–acceptor arrangement with
this connectivity cannot align properly in the hinge

6838
K. W. Woods et al. / Bioorg. Med. Chem. 14 (2006) 6832–6846
binding site. Attempts to provide a possibility for the de-
sired interaction led to the introduction of an amino
group in the 3-position (64). The compound, however,
showed little Akt activity (Akt1 Ki = 1232 nM).
5. Experimental
5.1. General procedures
1H NMR spectra were recorded on a Varian Mercury
300 (300 MHz) spectrometer or a Varian Unity Inova
500 (500 MHz) spectrometer. Chemical shifts (d) are
reported in parts per million (ppm) relative to tetrameth-
ylsilane as an internal standard. When peak multiplici-
ties are given the following abbreviations are used: s,
singlet; d, doublet; t, triplet; q, quartet; m, multiplet;
br, broadened. Mass spectra were performed as follows:
ESI (electrospray ionization) was performed on a Finn-
igan SSQ7000 MS run as a flow injection acquisition;
DCI (desorption chemical ionization) was performed
on a Finnigan SSQ7000 MS using a direct exposure
probe with ammonia gas; APCI (atmospheric pressure
chemical ionization) was performed on a Finnigan Nav-
igator MS run as flow injection acquisition. Elemental
analyses were performed by Robertson Microlit, Madi-
son, NJ. Flash chromatography was carried out using
Merck 50–200 mm silica gel. All solvents and reagents
were obtained from commercial sources and used with-
out further purification, except where noted.
As was the case for the isoquinoline analogues, the
indazole analogues also seem to exhibit a balance
between geometric requirements and electronic
requirements. The cinnoline analogue (39, Akt1
Ki = 215 nM) was much less active than the isoquino-
line analogue (3, Akt1 Ki = 0.99 nM) even though the
hinge binding nitrogen atoms should be in the same
spatial orientation. Replacing the indazole (48, Akt1
Ki = 0.99 nM) with benztriazole (65) maintains a simi-
lar spatial relationship of the nitrogen atoms but re-
sults in a nearly 100-fold loss of Akt activity (Akt1
Ki = 92 nM).
The 3-methylindazole compound 4 was chosen for fur-
ther investigation. Not only was this compound the
most potent of the inhibitors against Akt1 (Akt1
Ki = 0.16 nM) but it also showed a favorable selectiv-
ity profile (Table 2). There is little selectivity within
the Akt isoforms. Compound 4 is least selective
against closely related kinases in the AGC family,
nonetheless, it is 40-fold selective against PKA. The
inhibitor 4 also demonstrated cellular growth inhibi-
tion activity against MiaPaCa cells (Soft Agar
5.2. Chemistry
5.2.1.
(S)-[2-(5-Bromo-pyridin-3-yloxy)-1-(1H-indol-3-
EC50 = 44 nM;
EC50 = 300 nM).
MTT
EC50 = 100 nM;
GSK3-P
ylmethyl)-ethyl]-carbamic acid tert-butyl ester (7). A
solution of 3-bromo-5-hydroxypyridine34 (2.0 g,
11.5 mmol), L-Boc-tryptophanol (3.67 g, 12.6 mmol),
and triphenylphosphine (4.53 g, 17.3 mmol) in 50 mL
THF at 0 ꢁC was treated dropwise with DEAD (3.01 g,
17.3 mmol). The reaction was warmed to room tempera-
ture and stirred overnight. The reaction mixture was con-
centrated and purified by flash column chromatography
on silica gel with 20% EtOAc/hexane to provide the de-
sired product 7 (4.55 g, 89%). 1H NMR (300 MHz,
DMSO-d6) d ppm 10.8 (br s, 1H), 8.27 (m, 2H), 7.65
(m, 1H), 7.56 (d, J = 8 Hz, 1H), 7.33 (d, J = 8 Hz, 1H),
6.9–7.1 (m, 4H), 4.0 (M, 2H), 2.9–3.0 (m, 3H), 1.35 (s,
9H). MS (DCI/NH3): m/z (M+H)+ 446, 448.
Although the 3-methylindazole compound 4 suffered
from a short half-life (t1/2 = 0.6 h in mouse) and no
oral bioavailability, it was examined for in vivo effica-
cy in several mouse tumor models. The compound was
dosed subcutaneously at 7.5 and 15 mg/kg/day for 14
days. Dosing was limited to 14 days due to skin irrita-
tion at the injection site. The inhibitor was found to
significantly slow the growth of the tumors. The de-
tails of these tumor studies have been previously
reported.22
4. Conclusion
5.2.2. (S)-[1-(1H-Indol-3-ylmethyl)-2-(5-trimethylstanna-
nyl-pyridin-3-yloxy)-ethyl]-carbamic acid tert-butyl ester
(8). A solution of 7 (1 g, 2.23 mmol) in DMA (15 mL)
was treated with hexamethylditin (1.8 mL, 5.6 mmol)
and Pd(PPh3)4 (0.4 g, 0.2 mmol). The reaction mixture
was heated to 75 ꢁC for 1.5 days. The mixture was added
to water and extracted three times with ethyl acetate.
The combined extracts were concentrated and the resi-
due was purified by flash column chromatography on
silica gel with 1:1 hexanes/ethyl acetate to provide the
We have prepared a series of heteroaryl-pyridine con-
taining inhibitors of Akt leading to the discovery of
the indazole-pyridine 4. These compounds bind in
the ATP binding site, are potent, ATP competitive,31
and reversible inhibitors of Akt activity.32 Compound
4 is highly potent against Akt1 (Ki = 0.16 nM). No
selectivity amongst the Akt isoforms is observed for
this analogue, but it shows good selectivity against a
panel of other kinases. It is least selective for other
members of the AGC family of kinases but is none-
theless 40-fold selective for Akt over closely related
PKA. The compound shows cellular activity and sig-
nificantly slows tumor growth in vivo when dosed
subcutaneously. The in vivo dosing, however, is limit-
ed as a result of severe irritation at the injection site.
Discussions of oral bioavailability and half-life issues
are addressed in other communications from our
laboratories.33
1
desired product 8 (0.4 g, 34 %). H NMR (300 MHz,
DMSO-d6) d ppm 10.8 (br s, 1H), 8.17 (d, J = 3 Hz,
1H), 8.15 (s, 1H), 7.54 (d, J = 8 Hz, 1H), 7.38 (s, 1H),
7.33 (d, J = 8 Hz, 1H), 6.9–7.1 (m, 4H), 4.0 (m, 2H),
2.9–3.0 (m, 3H), 1.36 (s, 9H), 0.29 (s, 9H). MS (DCI/
NH3): m/z (M+H)+ 528, 530, 532.
5.2.3. (R)-[2-(5-Bromo-pyridin-3-yloxy)-1-(1H-indol-3-
ylmethyl)-ethyl]-carbamic acid tert-butyl ester. The
R-enantiomer of 7 (required for the synthesis of 49) is

K. W. Woods et al. / Bioorg. Med. Chem. 14 (2006) 6832–6846
6839
prepared in the same manner described for 7 using D-
Boc-tryptophanol in place of the L-Boc-tryptophanol.
1H NMR (300 MHz, DMSO-d6) d ppm 10.8 (br s,
1H), 8.27 (m, 2H), 7.65 (m, 1H), 7.56 (d, J = 8 Hz,
1H), 7.33 (d, J = 8 Hz, 1H), 6.9–7.1 (m, 4H), 4.0 (M,
2H), 2.9–3.0 (m, 3H), 1.35 (s, 9H). MS (DCI/NH3): m/
z (M+H)+ 446, 448.
5.2.8. 6-Bromo-4-chloro-cinnoline (14). A solution of 13
(0.4 g, 1.8 mmol) in POCl3 (2.5 mL) was heated to
100 ꢁC for 2 h, then poured slowly onto ice. The
aqueous solution was cooled to 0 ꢁC and adjusted to
pH 5–7 with 50% NaOH. The solution was extracted
with ethyl acetate (2·), and the combined organic layers
were concentrated. The residue was purified by flash col-
umn chromatography on silica gel with 4:1 hexanes/
EtOAc to provide the desired product 14 (0.190 g,
5.2.4. (R)-[1-(1H-Indol-3-ylmethyl)-2-(5-trimethylstanna-
nyl-pyridin-3-yloxy)-ethyl]-carbamic acid tert-butyl ester.
The R-enantiomer of 8 is prepared in the same manner
described for 8 using D-Boc-tryptophanol pyridyl ether
1
43%). H NMR (300 MHz, DMSO-d6) d ppm 9.64 (s,
1H), 8.51 (d, J = 9 Hz, 1H), 8.42 (d, J = 2 Hz, 1H),
8.22 (dd, J = 9, 2 Hz, 1H). MS (ESI): m/z (M+H)+
243, 245, 247.
1
in place of example 7. H NMR (300 MHz, DMSO-d6)
d ppm 10.8 (br s, 1H), 8.17 (d, J = 3 Hz, 1H), 8.15 (s,
1H), 7.54 (d, J = 8 Hz, 1H), 7.38 (s, 1H), 7.33 (d,
J = 8 Hz, 1H), 6.9–7.1 (m, 4H), 4.0 (m, 2H), 2.9–3.0
(m, 3H), 1.36 (s, 9H), 0.29 (s, 9H). MS (DCI/NH3):
m/z (M+H)+ 528, 530, 532.
5.2.9. (6-Bromo-cinnolin-4-yl)-hydrazine (15). A solution
of 14 (2.6 g, 10.6 mmol) in ethanol (70 mL) was treated
with hydrazine monohydrate (3 mL, 90% solution), stir-
red at room temperature for 3 days, and filtered. The
solid was washed with water (50 mL) and diethyl ether
(50 mL) and dried under vacuum to provide the desired
product 15 (2.5 g, 100%). The material was carried on
with no additional purification.
5.2.5. N-(2-Acetyl-phenyl)-acetamide (11). A solution of
20-aminoacetophenone (5.0 g, 37 mmol) in dichloro-
methane (150 mL) at room temperature was treated with
triethylamine (5.3 mL, 40 mmol) and acetyl chloride
(3.2 mL, 45 mmol). The reaction mixture was stirred at
rt for 3 h. The reaction mixture was diluted with EtOAc
and washed with water. The aqueous layer was extracted
with ethyl acetate (2·). The combined extracts were
rinsed with brine (1·), dried over MgSO4, and concen-
trated to provide the desired product 11 of sufficient
purity to carry on with no additional purification
(6.5 g, 100%).
5.2.10. 6-Bromo-cinnoline (16). A solution of 15 (3.5 g,
14 mmol) in water (50 mL) was heated to reflux then
treated dropwise with a solution of CuSO4 (2.8 g,
17.5 mmol) in water (20 mL). The reaction mixture
was heated at reflux for 2 h. The mixture was cooled
to room temperature and adjusted to pH 7 with saturat-
ed NaHCO3 (aq). The reaction mixture was extracted
with ethyl acetate (2·). The combined extracts were
rinsed with brine, dried over MgSO4, and concentrated.
The material was purified by flash column chromatogra-
phy on silica gel with 1:1 hexanes/EtOAc to provide the
5.2.6. N-(2-Acetyl-4-bromo-phenyl)-acetamide (12). A
solution of 11 (6.5 g, 37 mmol) in acetic acid
(100 mL) at room temperature was treated with Br2
(4 mL, 84 mmol) and stirred for 75 min. The reaction
mixture was poured into water (200 mL) and filtered.
The solid was washed with water (2·) and hexanes
(2·) then dissolved in diethyl ether. The Et2O solution
was washed with brine (1·), dried over MgSO4, and
concentrated to provide the desired product 12 (8.5 g,
89%). The product was carried on with no additional
purification.
1
desired product 16 (0.7 g, 24%). H NMR (300 MHz,
DMSO-d6) d ppm 9.41 (d, J = 6 Hz, 1H), 8.51 (d,
J = 9 Hz, 1H), 8.26 (d, J = 2 Hz, 1H), 8.21 (d,
J = 6 Hz, 1H), 7.98 (dd, J = 9, 2 Hz, 1H). MS (ESI):
m/z (M+H)+ 209, 211.
5.2.11. 6-Bromo-1-chloro-isoquinoline (18). A solution of
6-bromo-1-hydroxyisoquinoline35 (9.205 g, 41.0 mmol)
in POCl3 (100 mL) was heated at 100 ꢁC for 4 h. The
reaction mixture was concentrated to dryness. The resi-
due was dissolved in EtOAc and the organic layer was
washed successively with 5% NaHCO3, water, and
brine, dried over MgSO4, and concentrated. The residue
was chromatographed on silica gel eluting with 30%
CH2Cl2/hexane to give the desired compound 18
(6.176 g, 62%). 1H NMR (300 MHz, CDCl3) d ppm
8.30 (d, J = 6 Hz, 1H), 8.22 (d, J = 9 Hz, 1H), 8.04 (d,
J = 2 Hz, 1H), 7.77 (dd, J = 9, 2 Hz, 1H), 7.52 (d,
J = 6 Hz, 1H). MS (DCI/NH3): m/z (M+H)+ 242, 244,
246.
5.2.7. 6-Bromo-1H-cinnolin-4-one (13). A solution of 12
(6.28 g, 24.4 mmol) in THF (75 mL) was treated with
concentrated HCl (aq) (15 mL) and water (15 mL).
The reaction was heated at reflux for 1 h then concen-
trated to remove the THF. The aqueous solution was
treated with additional water (5 mL) and concd HCl
(5 mL). The solution was cooled to 0 ꢁC, then treated
with a solution of NaNO2 (1.85 g, 26.84 mmol) in water
(10 mL) in five portions. The reaction mixture was
warmed to room temperature gradually over a 2 h peri-
od then stirred overnight at room temperature. The
reaction mixture was heated to reflux for 6 h and fil-
tered. The resulting solid was washed with water
(50 mL) and diethyl ether (50 mL) then dried under vac-
5.2.12. 6-Bromo-isoquinolin-1-ylamine (19). A mixture of
the chloride 18 (264 mg, 1.09 mmol), acetamide (1.3 g),
and K2CO3 (0.45 g) was heated at 180 ꢁC for 5 h. After
cooling to rt, the mixture was dissolved in ethyl acetate
and washed successively with water and brine, dried
over MgSO4, and concentrated. The residue was chro-
matographed on silica gel eluting with CH2Cl2/MeOH/
NH4OH (100:5:0.5) to give the desired compound 19
1
uum to provide the desired product 13 (3.0 g, 54%). H
NMR (300 MHz, DMSO-d6) d ppm 13.65 (br s, 1H),
8.12 (d, J = 2 Hz, 1H), 7.94 (dd, J = 9, 2 Hz, 1H), 7.82
(s, 1H), 7.57 (d, J = 9 Hz, 1H). MS (ESI): m/z (M+H)+
225, 227.

6840
K. W. Woods et al. / Bioorg. Med. Chem. 14 (2006) 6832–6846
(159 mg, 65%). 1H NMR (300 MHz, CDCl3) d ppm 7.96
(d, J = 6 Hz, 1H), 7.88 (d, J = 2 Hz, 1H), 7.68 (d,
J = 9 Hz, 1H), 7.57 (dd, J = 9, 2 Hz, 1H), 6.96 (d,
J = 6 Hz, 1H), 5.4 (br s, 2H). MS (DCI/NH3): m/z
(M+H)+ 223, 225.
over ice, and filtered. The solid was recrystallized from
H2O/methanol to provide the desired product 23
1
(3.7 g, 38%). H NMR (300 MHz, DMSO-d6) d ppm
13.25 (br s, 1H), 8.05 (s, 1H), 8.00 (s, 1H), 7.4–7.5 (m,
2H). MS (DCI/NH3): m/z (M+H)+ 197, 199.
5.2.13. (6-Bromo-isoquinolin-1-yl)-bis-carbamic acid tert-
butyl ester (20). A solution of 19 (616 mg, 2.76 mmol),
Boc2O (1.81 g), DMAP (67 mg), and triethylamine
(1.15 mL) in acetonitrile (15 mL) was stirred at rt for
2 h. The reaction mixture was concentrated and the res-
idue was chromatographed on silica gel eluting with
30% EtOAc/hexane to give the desired compound 20
(1.18 g, 71%). 1H NMR (300 MHz, CDCl3) d ppm
8.45 (d, J = 6 Hz, 1H), 8.06 (d, J = 2 Hz, 1H), 7.83 (d,
J = 9 Hz, 1H), 7.71 (dd, J = 9, 2 Hz, 1H), 7.57 (d,
J = 6 Hz, 1H) 1.32 (s, 18H). MS (DCI/NH3): m/z
(M+H)+ 423, 425.
5.2.18. 1-(5-Bromo-2-fluoro-phenyl)-ethanone (25) (R =
CH3)
5.2.18.1. Step 1. A solution of 5-bromo-2-fluorobenz-
aldehyde (24.75 g; 122 mmol) in Et2O (125 mL) at 0 ꢁC
was treated with 3.0 M MeMgBr in Et2O (43 mL,
129 mmol). The mixture was stirred for 30 min. then care-
fully diluted with water and acidified with 10% HCl (aq).
The aqueous layer was extracted with Et2O. The com-
bined extracts were rinsed successively with 10% HCl
(aq), water, and brine, dried (MgSO4), and evaporated
to give 1-(5-bromo-2-fluorophenyl)ethanol (26.6 g;
99%) of sufficient purity to carry on to the next step.
5.2.14. (S)-5-[2-tert-Butoxycarbonylamino-3-(1H-indol-
3-yl)-propoxy]-nicotinic acid (21). A solution of 7
(1.30 g, 3.02 mmol) and PdCl2Ædppf (123 mg) in 12 mL
1:1 THF/water was heated at 100 ꢁC under CO (800 psi)
for 19 h. The reaction mixture was cooled to room tem-
perature and diluted with water. The mixture was
extracted with dichloromethane (3·) and the combined
extracts were washed with water, dried (MgSO4), and
concentrated to provide the desired product 21 (912 mg,
76%) that was carried on with no further purification.
5.2.18.2. Step 2. A solution of 1-(5-bromo-2-fluor-
ophenyl)ethanol (26.6 g; 121 mmol) and manganese(IV)
oxide (53 g; 610 mmol) in p-dioxane (500 mL) was heat-
ed at reflux for 4 h. The reaction mixture was cooled and
filtered through Celiteꢂ. The filtrate was evaporated and
purified by flash chromatography (5–10% Et2O/hexane)
to yield the desired product 25 (R = CH3) as a nearly
colorless oil that solidified upon standing (20.5 g;
78%). 1H NMR (300 MHz, CDCl3) d ppm 7.99 (dd,
J = 6, 3 Hz, 1H), 7.60–7.64 (m, 1H), 7.0–7.1 (m, 1H),
2.63 (s, 3H). MS (DCI/NH3): m/z (M+H)+ 217, 219.
5.2.15. (S)-{1-(1H-Indol-3-ylmethyl)-2-[5-(pyridin-4-yl
carbamoyl)-pyridin-3-yloxy]-ethyl}-carbamic acid tert-
butyl ester (Boc-protected 45). A solution of 21 (410 mg,
1.0 mmol), 4-aminopyridine (100 mg, 1.0 mmol), EDC
(960 mg), and HOBt (680 mg) in DMF (10 mL) was
stirred at room temperature overnight. The reaction mix-
ture was diluted with dichloromethane, washed with
water, dried (MgSO4), and concentrated. The residue
was purified by flash column chromatography on silica
gel with ethyl acetate/methanol (8:1) to provide the
desired product (87 mg, 18%). MS (DCI/NH3): m/z
(M+H)+ 488.
5.2.19. 5-Bromo-3-methyl-1H-indazole (26) (R = CH3).
A solution of 25 (R = CH3) (10 g; 46 mmol) in 25 mL of
hydrazine monohydrate was heated at reflux for 9 h.
The reaction mixture was poured over ice and the result-
ing precipitate was collected. The product was purified
by flash chromatography (1:1 Et2O/hexane) to give the
desired indazole 26 (R = CH3) as a white solid (5.8 g;
1
60%). H NMR (300 MHz, DMSO-d6) d ppm 12.8 (br
s, 1H), 7.95 (s, 1H), 7.43 (s, 2H), 2.47 (s, 3H). MS
(DCI/NH3): m/z (M+H)+ 211, 213.
5.2.20. 5-Bromo-1,3-dimethyl-1H-indazole (27) (R =
CH3). To a solution of 60% NaH (115 mg; 2.84 mmol)
in 10 mL DMF was added indazole 26 (R = CH3)
(500 mg; 2.37 mmol). After 15 min at rt iodomethane
(465 mg; 3.21 mmol) was added and the reaction mix-
ture was stirred for 2 h. The reaction mixture was treat-
ed with water and extracted into EtOAc (3·). The
combined extracts were rinsed with brine (2·), dried
over MgSO4, and evaporated. The product was purified
by flash chromatography (1:1 Et2O/hexane) to give the
desired N-methylindazole 27 (R = CH3) as a white solid
(360 mg; 67%). 1H NMR (300 MHz, CDCl3) d ppm 7.79
(d, J = 2 Hz, 1H), 7.43 (dd, J = 9, 2 Hz, 1H), 7.21 (d,
J = 9 Hz, 1H), 3.98 (s, 3H), 2.53 (s, 3H). MS (DCI/
NH3): m/z (M+H)+ 225, 227.
5.2.16. (S)-5-[2-Amino-3-(1H-indol-3-yl)-propoxy]-N-
pyridin-4-yl-nicotinamide (45). A solution of Boc-pro-
tected 45 (85 mg, 0.17 mmol) in dichloromethane
(20 mL) at room temperature was treated with 4 N
HCl in dioxane (5 mL), stirred for 2 h, and concentrat-
ed. The residue was dissolved in water (1.5 mL) and
lyophilized to provide the desired product 45 as the
dihydrochloride salt (25 mg, 31%). 1H NMR
(300 MHz, DMSO-d6) d 11.32 (br s, 1H), 11.04 (br s,
1H), 8.83 (d, J = 1.4 Hz, 1H), 8.69 (d, J = 6.8 Hz, 2H),
8.59 (d, J = 2.7 Hz, 1H), 8.15 (br s, 2H), 8.08 (d,
J = 6.8 Hz, 2H), 7.85 (dd, J = 2.7, 1.4 Hz, 1H), 7.61 (d,
J = 7.8 Hz, 1H), 7.38 (d, J = 8.12 Hz, 1H), 7.29 (d,
J = 2.7 Hz, 1H), 7.10 (m, 1H), 7.01 (m, 1H), 4.33 (m,
1H), 4.16 (m, 1H), 3.87 (m, 1H), 3.16 (m, 2H). MS
(DCI/NH3): m/z (M+H)+ 388.
5.2.21. 5-Bromo-2-fluoro-benzoic acid (28). A solution of
5-bromo-2-fluorobenzaldehyde (810 mg; 4.0 mmol) in
5 mL MeOH was treated with 3 mL of 15% NaOH
(aq) and 5 mL of 30% H2O2. The reaction mixture was
stirred at rt for 2 h. The mixture was acidified with
5.2.17. 5-Bromo-1H-indazole (23). A mixture of 5-bro-
mo-2-fluorobenzaldehyde (10 g, 49.2 mmol) and 98%
hydrazine (20 mL) was heated at reflux for 5 h, poured

K. W. Woods et al. / Bioorg. Med. Chem. 14 (2006) 6832–6846
6841
10% HCl (aq) and the resulting precipitate was collected,
rinsed with water, and dried to yield the desired product
28 (670 mg; 77%) which was carried on with no further
purification. 1H NMR (300 MHz, DMSO-d6) d ppm
13.6 (br s, 1H), 7.96 (dd, J = 6, 3 Hz, 1H), 7.8-7.9 (m,
1H), 7.3-7.4 (m, 1H). MS (DCI/NH3): m/z (M+H)+
219, 221.
1.54 mmol) then heated at 110 ꢁC for 4 h. The reaction
mixture was partitioned between brine and EtOAc, fil-
tered through Celiteꢂ, and extracted with EtOAc. The
extracts were rinsed with brine and dried over MgSO4.
The product was purified by flash chromatography
(1:1 EtOAc/hexane) to provide the desired product 33
1
(265 mg; 49%). H NMR (300 MHz DMSO-d6), d ppm
10.8 (br s, 1H), 8.54 (s, 1H), 8.35–8.4 (m, 1H), 8.31–
8.33 (m, 1H), 8.10–8.20 (m, 1H), 7.55–7.70 (m, 3H),
7.33 (d, J = 8 Hz, 1H), 7.15–7.20 (m, 1H), 6.90–7.10
(m, 3H), 4.1–4.15 (m, 2H), 2.8–3.05 (m, 3H), 1.36 (s,
9H). MS (ESI): m/z (M+H)+ 487.
5.2.22. (5-Bromo-2-fluoro-phenyl)-(1H-pyrrol-2-yl)-meth-
anone (30). The carboxylic acid 28 (665 mg; 3.0 mmol)
was dissolved in thionyl chloride (7 mL) and heated at
reflux for 2 h. The reaction mixture was concentrated
and azeotroped with toluene. The resulting acid chloride
29 and pyrrole (203 mg; 3.0 mmol) were taken up in 1,2-
dichloroethane (15 mL) and cooled to 0 ꢁC. AlCl3
(420 mg; 3.15 mmol) was added portionwise then stirred
overnight while gradually warming to rt. The reaction
mixture was poured over ice and acidified with 1 N
HCl then stirred at rt for 1.5 h. The solution was extract-
ed with CH2Cl2 (3·). The combined extracts were rinsed
with water and saturated NaHCO3 (aq), dried over
Na2SO4, and evaporated. The product 30 was isolated
by flash chromatography (10% EtOAc/hexane) as a pur-
5.2.27. (S)-[2-[5-(3-Amino-1H-indazol-5-yl)-pyridin-3-yl-
oxy]-1-(1H-indol-3-ylmethyl)-ethyl]-carbamic acid tert-
butyl ester (Boc-protected 59). A mixture of 33 (120 mg,
0.25 mmol) in 5 mL of 98% hydrazine was heated to reflux
for 5 h then poured over ice. The solution was extracted
with ethyl acetate, dried over MgSO4, and concentrated.
Purification by flash chromatography (7% MeOH/
CH2Cl2) provided the desired product Boc-protected 59
1
(103 mg, 84%). H NMR (300 MHz, DMSO-d6) d ppm
11.5 (br s, 1H), 10.8 br s, 1H), 8.48 (d, J = 2 Hz, 1H),
8.20 (d, J = 3 Hz, 1H), 8.10 (s, 1H), 7.55–7.60 (m, 3H),
7.30–7.35 (m, 3H), 7.15–7.18 (m, 1H), 6.90–7.05 (m,
2H), 5.43 (s, 2H), 4.05–4.15 (m, 2H), 2.90–3.05 (m, 3H),
1.36 (s, 9H). MS (ESI): m/z (M+H)+ 499.
1
ple solid (252 mg; 31%). H NMR (300 MHz, DMSO-
d6) d ppm 12.2 (br s, 1H), 7.7–7.8 (m, 2H), 7.35 (t,
J = 9 Hz, 1H), 7.25–7.3 (m, 1H), 6.6–6.7 (m, 1H), 6.2–
6.3 (m, 1H). MS (DCI/NH3): m/z (M+H)+ 268, 270.
5.2.23. 5-Bromo-3-(1H-pyrrol-2-yl)-1H-indazole (31).
Conversion of the ketone 30 to the indazole 31 was car-
ried out as described for example 26. 1H NMR
(300 MHz, DMSO-d6) d ppm 13.1 (br s, 1H), 11.4 (br
s, 1H), 8.18 (s, 1H), 7.45–7.5 (m, 2H), 6.85–6.9 (m,
1H), 6.7–6.75 (m, 1H), 6.15–6.2 (m, 1H). MS (DCI/
NH3): m/z (M+H)+ 262, 264.
5.2.28.
(S)-5-{5-[2-Amino-3-(1H-indol-3-yl)-propoxy]-
pyridin-3-yl}-1H-indazol-3-ylamine (59). A solution of
Boc-protected 59 (95 mg; 0.19 mmol) in 5 mL CH2Cl2
was treated with 0.5 mL TFA and stirred at rt for 3 h.
The reaction mixture was concentrated and the product
was purified by reverse-phase HPLC on a C18 column
with 0–100% CH3CN/H2O/0.1%TFA to provide the de-
sired product 59 as a TFA salt (114 mg; 81%). 1H NMR
(300 MHz, DMSO-d6) d ppm 11.04 (s, 1 H) 11.92 (br s,
1H), 8.57 (d, J = 1.70 Hz, 1H), 8.32 (d, J = 2.71 Hz, 1H),
8.18 (m, 4H), 7.66 (s, 1H), 7.63 (d, J = 7.46 Hz, 1H),
7.42 (s, 1H), 7.38 (m, 1H), 7.30 (d, J = 2.37 Hz, 1H),
7.12 (m, 4H), 4.36 (m, 1H), 4.18 (dd, J = 10.51,
5.76 Hz, 1H), 3.84 (m, 1H), 3.17 (d, J = 7.12 Hz, 2H).
MS (ESI): m/z (M+H)+ 399. Anal. Calcd for
C23H22N6OÆ2.9TFA: C, 47.44; H, 3.44; N, 11.53. Found:
C, 47.87; H, 3.49; N, 11.19.
5.2.24. 5-Bromo-1H-benzotriazole (35). 4-Bromo-1,2-
benzenediamine (262 mg; 1.4 mmol) in 4 mL of 10%
H2SO4 (aq) was treated with an aqueous solution of
NaNO2 (120 mg; 1.7 mmol in 1 mL H2O). A tan precip-
itate formed almost immediately. The reaction mixture
was stirred for 30 min. The mixture was diluted with
water and extracted into EtOAc (3·). The combined ex-
tracts were rinsed with brine, dried over Na2SO4, and
evaporated. The product was isolated by flash chroma-
tography (5% MeOH/CH2Cl2). The product was ob-
tained as a tan solid (200 mg; 72%). 1H NMR
(300 MHz, DMSO-d6) d ppm 15.95 (br s, 1H), 8.21 (s,
1H), 7.92 (d, J = 9 Hz, 1H), 7.59 (d, J = 9 Hz, 1H).
MS (DCI/NH3): m/z (M+H)+ 196, 198.
5.2.29. (S)-1-(1H-Indol-3-ylmethyl)-2-(5-naphthalen-2-yl-
pyridin-3-yloxy)-ethylamine (37). Stille reaction with 2-
bromonaphthalene and stannyl material 8 (as described
for 33) followed by deprotection of the Boc group with
TFA (as described for 59) yielded 37. 1H NMR
(500 MHz, DMSO-d6) d ppm 11.02 (s, 1H), 8.74 (s,
1H), 8.38 (s, 1H), 8.30 (s, 1H), 8.18–8.21 (m, 2H), 8.04
(d, J = 8 Hz, 1H), 7.97–8.01 (m, 2H), 8.85 (d,
J = 8 Hz, 1H), 7.81 (s, 1H), 7.62 (d, J = 8 Hz, 1H),
7.50–7.58 (m, 1H), 7.35–7.39 (m, 1H), 7.23–7.31 (m,
1H), 7.08–7.12 (m, 1H), 6.96–7.03 (m, 2H), 4.18–4.41
(m, 2H), 3.82–3.87 (m, 1H), 3.17–3.21 (m, 2H). MS
(ESI): m/z (M+H)+ 394.
5.2.25. 5-Bromo-benzotriazole-1-carboxylic acid tert-bu-
tyl ester (36). The Boc group was introduced onto the
benztriazole 35 nitrogen as described by Katritzky
et al.28
5.2.26.
yloxy]-1-(1H-indol-3-ylmethyl)-ethyl]-carbamic
(S)-[2-[5-(3-Cyano-4-fluoro-phenyl)-pyridin-3-
acid
tert-butyl ester (33). A solution of 5-bromo-2-fluor-
obenzonitrile (246 mg; 1.23 mmol) and stannyl material
8 (595 mg; 1.12 mmol) in DMF (10 mL) was treated
with Pd2(dba)3 (113 mg; 0.123 mmol),tri-o-tolylphos-
phine (80 mg; 0.246 mmol), and triethylamine (156 mg;
5.2.30. (S)-2-(1H-indol-3-yl)-1-{[(5-isoquinolin-6-ylpyri-
din-3-yl)oxy]methyl}ethylamine (3). Stille reaction with
6-bromoisoquinoline and stannyl material 8 (as de-

6842
K. W. Woods et al. / Bioorg. Med. Chem. 14 (2006) 6832–6846
scribed for 33) followed by deprotection of the Boc
group with TFA (as described for 59) yielded 3. 1H
NMR (300 MHz, DMSO-d6) d ppm 11.02 (br s, 1H),
9.52 (s, 1H), 8.76 (d, J = 3 Hz, 1H), 8.62 (d, J = 8 Hz,
1H), 8.44–8.46 (m, 2H), 8.38 (d, J = 9 Hz, 1H), 8.11–
8.20 (m, 3H), 8.04–8.08 (m, 1H), 7.83–7.86 (m, 1H),
7.62 (d, J = 9 Hz, 1H), 7.37–7.40 (m, 1H), 7.31 (d,
J = 3 Hz, 1H), 7.08–7.12 (m, 1H), 6.99–7.03 (m, 1H),
4.37–4.41 (m, 1H), 4.18–4.23 (m, 1H), 3.86–3.91 (m,
1H), 3.16–3.20 (m, 2H). MS (ESI): m/z (M+H)+ 395.
Anal. Calcd for C25H22N4OÆ2TFAÆH2O: C, 49.35; H,
3.61; N, 7.43; F, 22.67. Found: C, 49.04; H, 3.55; N,
7.42; F, 22.28.
deprotection of the Boc group with TFA (as described
for 59) yielded 41. H NMR (300 MHz, DMSO-d6) d
1
ppm 8.77 (s, 1H), 8.55 (d, J = 8.6 Hz, 1H), 8.45 (d,
J = 2.5 Hz, 1H), 8.20 (d, J = 1.5 Hz, 1H), 8.05 (dd,
J = 8.6, 1.8 Hz, 1H), 7.83 (dd, J = 1.8, 2.5 Hz, 1H),
7.61 (m, 2H), 7.38 (d, J = 7.1 Hz, 1H), 7.24 (s, 1H),
7.12 (m, 1H), 7.02 (m, 1H), 4.44 (dd, J = 10.4, 3.1 Hz,
1H), 4.30 (dd, J = 10.4, 5.8 Hz, 1H), 4.01 (m, 1H),
3.32 (m, 2H). MS (DCI/NH3): m/z (M+H)+ 410.
5.2.35.
6-{5-[(S)-2-amino-3-(1H-indol-3-yl)-propoxy]-
pyridin-3-yl}-1-chloro-isoquinoline (42). Stille reaction
with 6-bromo-1-chloroisoquinoline (18) and stannyl
material 8 (as described for 33) followed by deprotection
of the Boc group with TFA (as described for 59) yielded
5.2.31. (S)-2-(1H-indol-3-yl)-1-{[(5-quinolin-6-ylpyridin-
3-yl)oxy]methyl}ethylamine (38). Stille reaction with 6-
bromoquinoline and stannyl material 8 (as described
for 33) followed by deprotection of the Boc group with
TFA (as described for 59) yielded 38. 1H NMR
(500 MHz, DMSO-d6) d ppm 11.02 (s, 1H), 8.97-9.00
(m, 1H), 8.74 (d, J = 3 Hz, 1H), 8.50–8.54 (m, 1H),
8.39–8.42 (m, 2H), 8.18–8.23 (m, 3H), 8.13–8.17 (m,
1H), 7.81–7.83 (m, 1H), 7.61–7.66 (m, 2H), 7.39 (d,
J = 8 Hz, 1H), 7.31 (d, J = 3 Hz, 1H), 7.07–7.10 (m,
1H), 6.99–7.02 (m, 1H), 4.38–4.41 (m, 1H), 4.21–4.24
(m, 1H), 3.79–3.83 (m, 1H), 3.16–3.19 (m, 2H). MS
(ESI): m/z (M+H)+ 395.
1
42. H NMR (300 MHz, DMSO-d6) d ppm 11.03 (br s,
1H), 8.75 (d, J = 1.6 Hz, 1H), 8.47 (d, J = 1.6 Hz, 1H),
8.45 (d, J = 5.6 Hz, 1H), 7.84 (m, 1H), 7.62 (d,
J = 8.1 Hz, 1H), 7.39 (d, J = 8.1 Hz, 1H), 7.31 (d,
J = 2.2 Hz, 1H), 7.10 (m, 1H), 7.01 (m, 1H), 4.38 (dd,
J = 10.6, 3.1 Hz, 1H), 4.22 (dd, J = 10.4, 6.2 Hz, 1H),
3.88 (m, 1H), 3.18 (m, 2H). MS (ESI): m/z (M+H)+
429, 431.
5.2.36. (S)-1-(1H-Indol-3-ylmethyl)-2-(5-isoquinolin-5-yl-
pyridin-3-yloxy)-ethylamine (43). Stille reaction with 5-
bromoisoquinoline and stannyl material 8 (as described
for 33) followed by deprotection of the Boc group with
TFA (as described for 59) yielded 43. 1H NMR
(300 MHz, DMSO-d6) d ppm 11.02 (br s, 1H), 9.53 (s,
1H), 8.52 (d, J = 8 Hz, 1H), 8.49 (d, J = 4 Hz, 1H),
8.37 (d, J = 3 Hz, 1H), 8.30–8.34 (m, 1H), 8.15–8.19
(m, 2H), 7.84–7.88 (m, 2H), 7.68 (d, J = 8 Hz, 1H),
7.56–7.60 (m, 2H), 7.47 (d, J = 8 Hz, 1H), 7.28 (d,
J = 4 Hz, 1H), 7.60–7.12 (m, 1H), 6.94–6.99 (m, 1H),
4.12–4.32 (m, 2H), 3.82–3.87 (m, 1H), 3.13–3.17 (m,
2H). MS (ESI): m/z (M+H)+ 395.
5.2.32. (S)-2-[(5-cinnolin-6-ylpyridin-3-yl)oxy]-1-(1H-indol-
3-ylmethyl)ethylamine (39). Stille reaction with 6-bro-
mocinnoline (16) and stannyl material 8 (as described
for 33) followed by deprotection of the Boc group with
TFA (as described for 59) yielded 39. 1H NMR
(300 MHz, DMSO-d6) d ppm 11.04 (s, 1H), 9.43 (d,
J = 6 Hz, 1H), 8.78 (d, J = 2 Hz, 1H), 8.60 (d,
J = 8 Hz, 1H), 8.45–8.49 (m, 2H), 8.30–8.34 (m, 1H),
8.26 (d, J = 6 Hz, 1H), 8.21–8.25 (m, 2H), 7.89 (t,
J = 2 Hz, 1H), 7.63 (d, J = 8 Hz, 1H), 7.39 (d,
J = 8 Hz, 1H), 7.31 (d, J = 2 Hz, 1H), 7.08–7.12 (m,
1H), 7.01–7.04 (m, 1H), 4.38–4.42 (m, 1H), 4.22–4.26
(m, 1H), 3.83–3.88 (m, 1H), 3.17–3.20 (m, 2H). MS
(ESI): m/z (M+H)+ 396.
5.2.37.
(S)-2-{[5-(1H-indazol-5-yl)pyridin-3-yl]oxy}-1-
(1H-indol-3-ylmethyl)-ethylamine (48). Stille reaction
with 5-bromoindazole (23) and stannyl material 8 (as de-
scribed for 33) followed by deprotection of the Boc
1
group with TFA (as described for 59) yielded 48. H
5.2.33.
6-{5-[(S)-2-Amino-3-(1H-indol-3-yl)-propoxy]-
NMR (300 MHz, DMSO-d6) d ppm 13.22 (br s, 1H),
11.04 (br s, 1H), 8.62 (d, J = 2 Hz, 1H), 8.33 (d,
J = 3 Hz, 1H), 8.13–8.21 (m, 3H), 8.12 (s, 1H), 7.67–
7.72 (m, 3H), 7.64 (d, J = 8 Hz, 1H), 7.39 (d, J = 8Hz,
1H), 7.30 (d, J = 2 Hz, 1H), 7.06–7.13 (m, 1H), 6.98–
7.04 (m, 1H), 4.14–4.39 (m, 2H), 3.33–3.38 (m, 1H),
3.13–3.16 (m, 2H). MS (ESI): m/z (M+H)+ 384. Anal.
Calcd for C23H21N5OÆ2TFAÆH2O: C, 51.52; H, 4.00;
N, 11.13. Found: C, 51.80; H, 3.61; N, 11.03.
pyridin-3-yl}-2H-isoquinolin-1-one (40). Stille reaction
with 6-bromo-1-hydroxyisoquinoline (17) and stannyl
material 8 (as described for 33) followed by deprotection
of the Boc group with TFA (as described for 59) yielded
1
40. H NMR (300 MHz, DMSO-d6) d ppm 11.30 (br s,
1H), 11.04 (br s, 1H), 8.66–8.68 (m, 1H), 8.41 (d,
J = 3 Hz, 1H), 8.27 (d, J = 8 Hz, 1H), 8.17–8.20 (m,
2H), 8.02–8.03 (m, 1H), 7.76–7.81 (m, 2H), 7.62 (d,
J = 8 Hz, 1H), 7.38 (d, J = 8 Hz, 1H), 7.29–7.31 (m,
1H), 7.20–7.26 (m, 1H), 7.07–7.12 (m, 1H), 6.98–7.04
(m, 1H), 6.60 (d, J = 8 Hz, 1H), 4.14–4.39 (m, 2H),
3.33–3.38 (m, 1H), 3.13–3.16 (m, 2H). MS (ESI): m/z
(M+H)+ 411. Anal. Calcd for C25H22N4OÆ2TFA: C,
54.54; H, 3.78; N, 8.78. Found: C, 54.54; H, 4.00; N, 8.56.
5.2.38. (S)-2-([3,40]Bipyridinyl-5-yloxy)-1-(1H-indol-3-
ylmethyl)-ethylamine (44). Stille reaction with 4-bromo-
pyridine and stannyl material 8 (as described for 33) fol-
lowed by deprotection of the Boc group with TFA (as
described for 59) yielded 44. 1H NMR (300 MHz,
CD3OD) d ppm 8.85 (d, J = 6.8 Hz, 2H), 8.73 (d,
J = 1.7 Hz, 1H), 8.53 (d, J = 2.7 Hz, 1H), 8.20 (d,
J = 6.8 Hz, 2H), 7.84 (t, J = 1.7 Hz, 1H), 7.59 (d,
J = 7.8 Hz, 1H), 7.38 (d, J = 8.2 Hz, 1H), 7.24 (s, 1H),
7.12 (t, J = 6.8 Hz, 1H), 7.01 (t, J = 8.1 Hz, 1H), 4.44
5.2.34.
1-Amino-6-{5-[(S)-2-amino-3-(1H-indol-3-yl)-
propoxy]-pyridin-3-yl}-isoquinoline (41). Stille reaction
with 1-bis-Boc-amino-6-bromoisoquinoline (20) and
stannyl material 8 (as described for 33) followed by

K. W. Woods et al. / Bioorg. Med. Chem. 14 (2006) 6832–6846
6843
(dd, J = 10.8, 3.4 Hz, 1H), 4.30 (dd, J = 10.5, 5.8 Hz,
1H), 4.01 (m, 1H), 3.33 (m, 2H). MS (APCI): m/z
(M+H)+ 345. Anal. Calcd for C21H20N4OÆ2.7TFA: C,
48.61; H, 3.51; N, 8.59. Found: C, 48.69; H, 3.50; N,
8.46.
J = 7.5 Hz, 1H), 7.00 (t, J = 7.5 Hz, 1H), 4.26–4.46 (m,
2H), 3.78–3.88 (m, 1H), 3.19–3.22 (m, 2H), 2.56 (s,
3H). MS (ESI): m/z (M+H)+ 398. Anal. Calcd for
C24H23N5OÆ2.0HClÆ0.75H2O: C, 59.57; H, 5.52; N,
14.47, Cl, 14.65. Found: C, 59.73; H, 5.36; N, 14.32,
Cl, 14.74.
5.2.39. (S)-(4-(5-(2-Amino-3-(1H-indol-3-yl)-propoxy)-
pyridin-3-yl)-phenyl)-methanol (46). Stille reaction with
4-bromobenzyl alcohol and stannyl material 8 (as de-
scribed for 33) followed by deprotection of the Boc
5.2.43. (S)-2-[5-(1,3-Dimethyl-1H-indazol-5-yl)-pyridin-
3-yloxy]-1-(1H-indol-3-ylmethyl)-ethylamine (50). Stille
reaction with 5-bromo-1,3-dimethylindazole (27
R = CH3) and stannyl material 8 (as described for 33)
followed by deprotection of the Boc group with TFA
1
group with TFA (as described for 59) yielded 46. H
NMR (500 MHz, DMSO-d6) d ppm 11.03 (s, 1H), 8.51
(d, J = 1.56 Hz, 1H), 8.30 (d, J = 2.81 Hz, 1H), 7.64
(dd, J = 10.61, 8.42 Hz, 3H), 7.59 (m, 1H), 7.43 (d,
J = 8.42 Hz, 2H), 7.37 (d, J = 8.11 Hz, 1H), 7.28 (d,
J = 2.18 Hz, 1H), 7.09 (t, J = 7.02 Hz, 1H), 6.99 (t,
J = 7.02 Hz, 1H), 5.29 (s, 1H), 4.55 (s, 2H), 4.29 (m,
1H), 4.16 (dd, J = 10.29, 5.93 Hz, 1H), 3.72 (m, 1H),
3.15 (m, 4H). MS (ESI): m/z (M+H)+ 374.
1
(as described for 59) yielded 50. H NMR (300 MHz,
DMSO-d6) d ppm 11.04 (s, 1H) 8.66 (d, J = 1.70 Hz,
1H) 8.34 (d, J = 2.71 Hz, 1H) 8.18 (m, 3H) 8.08 (s,
1H) 7.73 (m, 2H) 7.63 (d, J = 7.80 Hz, 1H) 7.39 (d,
J = 7.80 Hz, 1H) 7.30 (d, J = 2.37 Hz, 1H) 7.10 (t,
J = 7.12 Hz, 1H) 7.01 (t, J = 7.46 Hz, 1H) 4.37 (m,
1H) 4.20 (dd, J = 10.51, 6.10 Hz, 1H) 4.00 (s, 3H) 3.86
(s, 1H) 3.18 (m, 2H) 2.54 (s, 3H). MS (ESI): m/z
(M+H)+ 412. Anal. Calcd for C25H25N5OÆ2.8TFA: C,
50.29; H, 3.83; N, 9.58. Found: C, 50.36; H, 3.84, N,
9.60.
5.2.40. 4-(5-{[(S)-2-amino-3-(1H-indol-3-yl)propyl]oxy}-
pyridin-3-yl)benzonitrile (47). Stille reaction with 4-bro-
mobenzonitrile and stannyl material 8 (as described
for 33) followed by deprotection of the Boc group with
TFA (as described for 59) yielded 47. 1H NMR
(300 MHz,DMSO-d6) d ppm 11.02 (s, 1H), 8.63 (d,
J = 1.9 Hz, 1H), 8.42 (d, J = 2.8 Hz, 1H), 8.21 (br s,
2H), 7.99-7.92 (m, 4H), 7.73 (t, J = 1.9 Hz, 1H), 7.61
(d, J = 8.1 Hz, 1H), 7.38 (d, J = 8.1 Hz, 1H), 7.29 (d,
J = 2.5 Hz, 1H), 7.10 (m, 1H), 7.01 (m, 1H), 4.36 (dd,
J = 10.6, 3.1 Hz, 1H), 4.19 (dd, J = 10.9, 5.9 Hz, 1H),
3.89–3.82 (m, 1H), 3.16 (d, J = 7.2 Hz, 2H). MS (DCI/
NH3): m/z (M+H)+ 369.
5.2.44. (S)-2-[5-(3-Ethyl-1H-indazol-5-yl)-pyridin-3-yloxy]
-1-(1H-indol-3-ylmethyl)-ethylamine (51). Stille reaction
with 5-bromo-3-ethylindazole (26 R = Et) and stannyl
material 8 (as described for 33) followed by deprotection
of the Boc group with TFA (as described for 59) yielded
1
51. H NMR (300 MHz, DMSO-d6) d ppm 12.80 (m,
1H) 11.04 (d, J = 2.03 Hz, 1H) 8.63 (d, J = 1.70 Hz,
1H) 8.33 (d, J = 2.71 Hz, 1H) 8.16 (m, 2H) 8.08 (s,
1H) 7.72 (m, 1H) 7.63 (m, 3H) 7.38 (d, J = 7.80 Hz,
1H) 7.30 (d, J = 2.37 Hz, 1H) 7.11 (t, J = 7.46 Hz, 1H)
7.01 (t, J = 7.46 Hz, 1H) 4.37 (m, 1H) 4.19 (dd,
J = 10.85, 6.10 Hz, 1H) 3.86 (m, 1H) 3.17 (m, 2H) 2.99
(q, J = 7.57 Hz, 2H) 1.35 (t, J = 7.63 Hz, 3H). MS
(ESI): m/z (M+H)+ 412. Anal. Calcd for
C25H25N5OÆ2.7TFA: C, 50.76; H, 3.88; N, 9.74. Found:
C, 51.09; H, 3.88; N, 9.66.
5.2.41. (S)-1-(1H-Indol-3-ylmethyl)-2-[5-(3-methyl-1H-
indazol-5-yl)-pyridin-3-yloxy]-ethylamine (4). Stille reac-
tion with 5-bromo-3-methylindazole (26 R = CH3) and
stannyl material 8 (as described for 33) followed by
deprotection of the Boc group with TFA (as described
for 59) yielded 4. The product obtained was converted
to the HCl salt. 1H NMR (300 MHz, DMSO-d6) d
ppm 12.8 (br s, 1H), 11.06 (s, 1H), 8.76 (s, 1H), 8.44
(m, 3H), 8.17 (s, 1H), 8.01 (s, 1H), 7.73 (dd, J = 9,
2 Hz, 1H), 7.66 (d, J = 8 Hz, 1H), 7.58 (d, J = 9 Hz,
1H), 7.38 (d, J = 8 Hz, 1H), 7.32 (d, J = 2 Hz, 1H),
7.10 (t, J = 7.5 Hz, 1H), 7.00 (t, J = 7.5 Hz, 1H), 4.26–
4.46 (m, 2H), 3.78–3.88 (m, 1H), 3.19–3.22 (m, 2H),
2.56 (s, 3H). MS (ESI): m/z (M+H)+ 398. Anal. Calcd
for C24H23N5OÆ2.25HCl: C, 59.62; H, 5.31; N, 14.60,
Cl, 16.63. Found: C, 59.62; H, 5.31; N, 14.28, Cl, 16.22.
5.2.45. (S)-2-[5-(3-Cyclopropyl-1H-indazol-5-yl)-pyridin-
3-yloxy]-1-(1H-indol-3-ylmethyl)-ethylamine (52). Stille
reaction with 5-bromo-3-cyclopropylindazole (26
R = cyclopropyl) and stannyl material 8 (as described
for 33) followed by deprotection of the Boc group with
TFA (as described for 59) yielded 52. 1H NMR
(300 MHz, DMSO-d6) d ppm 12.73 (m, 1H) 11.03 (d,
J = 2.03 Hz, 1H) 8.63 (d, J = 1.36 Hz, 1H) 8.33 (d,
J = 2.37 Hz, 1H) 8.19 (m, 2H) 8.12 (s, 1H) 7.73 (m,
1H) 7.65 (m, 2H) 7.56 (d, J = 8.48 Hz, 1H) 7.38 (d,
J = 8.14 Hz, 1H) 7.30 (d, J = 2.37 Hz, 1H) 7.10 (t,
J = 6.95 Hz, 1H) 7.01 (t, J = 7.46 Hz, 1H) 4.37 (dd,
J = 10.68, 3.22 Hz, 1H) 4.19 (dd, J = 10.68, 5.93 Hz,
1H) 3.86 (m, 1H) 3.15 (m, 2H) 2.36 (m, 1H) 1.02 (m,
4H). MS (ESI): m/z (M+H)+ 424. Anal. Calcd for
C26H25N5OÆ2.6TFA: C, 52.05; H, 3.86; N, 9.73. Found:
C, 52.03; H, 3.89; N, 9.69.
5.2.42. (R)-1-(1H-Indol-3-ylmethyl)-2-[5-(3-methyl-1H-
indazol-5-yl)-pyridin-3-yloxy]-ethylamine (49). Stille
reaction with 5-bromo-3-methylindazole (26 R = CH3)
and stannyl material (the R-enantiomer of 8) (as de-
scribed for 33) followed by deprotection of the Boc
group with TFA (as described for 59) yielded 49 as the
TFA salt that was subsequently converted to the HCl
1
salt. H NMR (300 MHz, DMSO-d6) d ppm 12.8 (br s,
1H), 11.06 (s, 1H), 8.76 (s, 1H), 8.44 (m, 3H), 8.17 (s,
1H), 8.01 (s, 1H), 7.73 (dd, J = 9, 2 Hz, 1H), 7.66 (d,
J = 8 Hz, 1H), 7.58 (d, J = 9 Hz, 1H), 7.38 (d,
J = 8 Hz, 1H), 7.32 (d, J = 2 Hz, 1H), 7.10 (t,
5.2.46. (S)-1-(1H-Indol-3-ylmethyl)-2-[5-(3-phenyl-1H-
indazol-5-yl)-pyridin-3-yloxy]-ethylamine (53). Stille
reaction with 5-bromo-3-phenylindazole (26 R = phen-
yl) and stannyl material 8 (as described for 33) followed

6844
K. W. Woods et al. / Bioorg. Med. Chem. 14 (2006) 6832–6846
by deprotection of the Boc group with TFA (as de-
scribed for 59) yielded 53. 1H NMR (300 MHz,
DMSO-d6) d ppm 13.45 (br s, 1H) 11.03 (s, 1H) 8.68
(d, J = 1.70 Hz, 1H) 8.36 (d, J = 2.71 Hz, 1H) 8.30 (s,
1H) 8.16 (m, 2H) 8.08 (s, 1H) 8.05 (s, 1H) 7.76 (m,
1H) 7.72 (s, 2H) 7.62 (d, J = 7.46 Hz, 1H) 7.43–7.55
(m, 3H) 7.39 (m, 1H) 7.30 (d, J = 2.37 Hz, 1H) 7.09 (t,
J = 7.46 Hz, 1H) 7.00 (t, J = 7.46 Hz, 1H) 4.38 (m,
1H) 4.19 (dd, J = 10.51, 5.76 Hz, 1H) 3.87 (m, 1H)
3.17 (m, 2H). MS (ESI): m/z (M+H)+ 460. Anal. Calcd
for C29H25N5OÆ3TFA: C, 52.44; H, 3.52; N, 8.74.
Found: C, 52.91; H, 3.68; N, 8.80.
(24) (prepared as described in Wrzeciono et al.27) and
stannyl material 8 (as described for 33) followed by
deprotection of the Boc group with TFA (as described
1
for 59) yielded 60. H NMR (300 MHz, DMSO-d6) d
ppm 12.04 (s, 1H) 11.03 (s, 1H) 8.62 (d, J = 1.36 Hz,
1H) 8.33 (d, J = 2.37 Hz, 1H) 8.17 (m, 2H) 8.07 (s,
1H) 7.73 (s, 1H) 7.61 (m, 2H) 7.45 (d, J = 8.82 Hz,
1H) 7.38 (d, J = 8.14 Hz, 1H) 7.30 (d, J = 2.37 Hz, 1H)
7.10 (t, J = 7.12 Hz, 1H) 7.01 (t, J = 6.95 Hz, 1H) 4.36
(m, 1H) 4.19 (m, 1H) 3.86 (s, 1H) 3.16 (m, 2H) 3.04
(s, 6H). MS (ESI): m/z (M+H)+ 427. Anal. Calcd for
C25H26N6OÆ3.5TFA: C, 46.55; H, 3.60; N, 10.18. Found:
C, 46.71; H, 3.65, N, 10.02.
5.2.47. (S)-1-(1H-Indol-3-ylmethyl)-2-[5-(3-thiophen-2-
yl-1H-indazol-5-yl)-pyridin-3-yloxy]-ethylamine
(55).
5.2.51. (S)-1-(1H-Indol-3-ylmethyl)-2-[5-(3-morpholin-4-
yl-1H-indazol-5-yl)-pyridin-3-yloxy]-ethylamine
Stille reaction with 5-bromo-3-(thiophen-2-yl)indazole
(26 R = thiophen-2-yl) and stannyl material 8 (as de-
scribed for 33) followed by deprotection of the Boc
(61).
Stille reaction with 5-bromo-3-(morpholin-4-yl)indazole
(prepared as described in Wrzeciono et al.27) and stannyl
material 8 (as described for 33) followed by deprotection
of the Boc group with TFA (as described for 59) yielded
61. 1H NMR (300 MHz, DMSO-d6) d ppm 12.21 (s, 1H)
11.03 (s, 1H) 8.65 (d, J = 1.70 Hz, 1H) 8.33 (d,
J = 2.71 Hz, 1H) 8.17 (m, 2H) 8.09 (s, 1H) 7.72 (m,
1H) 7.62 (m, 2H) 7.48 (d, J = 8.82 Hz, 1H) 7.38 (d,
J = 7.80 Hz, 1H) 7.30 (d, J = 2.37 Hz, 1H) 7.10 (t,
J = 7.46 Hz, 1H) 7.01 (t, J = 7.46 Hz, 1H) 4.35 (m,
1H) 4.19 (dd, J = 10.68, 5.93 Hz, 1H) 3.88 (m, 1H)
3.81 (m, 4H) 3.35 (m, 4H) 3.16 (d, J = 7.12 Hz, 2H).
MS (ESI): m/z (M+H)+ 469. Anal. Calcd for
C27H28N6O2Æ3.4TFA: C, 47.41; H, 3.70; N, 9.82. Found:
C, 47.10; H, 3.86; N, 9.95.
1
group with TFA (as described for 59) yielded 55. H
NMR (400 MHz, DMSO-d6) d ppm 13.39 (s, 1H) 11.06
(s, 1H) 8.67 (s, 1H) 8.34 (m, 4H) 7.91 (d, J = 2.76 Hz,
1H) 7.73 (m, 3H) 7.64 (d, J = 7.98 Hz, 1H) 7.59 (d,
J = 6.14 Hz, 1H) 7.39 (d, J = 7.98 Hz, 1H) 7.31 (d,
J = 2.15 Hz, 1H) 7.23 (dd, J = 5.22, 3.68 Hz, 1H) 7.10
(t, J = 7.06 Hz, 1H) 7.00 (t, J = 7.52 Hz, 1H) 4.38 (m,
1H) 4.23 (dd, J = 10.43, 5.83 Hz, 1H) 3.86 (br s, 1H)
3.21 (d, J = 7.06 Hz, 2H). MS (ESI): m/z (M+H)+ 466.
5.2.48. (S)-1-(1H-Indol-3-ylmethyl)-2-[5-(3-thiazol-2-yl-
1H-indazol-5-yl)-pyridin-3-yloxy]-ethylamine (56). Stille
reaction with 5-bromo-3-(thiazol-2-yl)indazole (26
R = thiazol-2-yl) and stannyl material 8 (as described
for 33) followed by deprotection of the Boc group with
TFA (as described for 59) yielded 56. 1H NMR
(500 MHz, DMSO-d6) d ppm 13.74 (s, 1H) 11.04 (s,
1H) 8.60 (s, 2H) 8.38 (d, J = 2.18 Hz, 1H) 8.28 (s, 2H)
8.04 (d, J = 3.43 Hz, 1H) 7.79 (d, J = 3.12 Hz, 1H)
7.78 (s, 2H) 7.73 (s, 1H) 7.64 (d, J = 7.80 Hz, 1H) 7.38
(d, J = 8.11 Hz, 1H) 7.31 (d, J = 1.56 Hz, 1H) 7.09 (t,
J = 7.49 Hz, 1H) 7.01 (t, J = 7.49 Hz, 1H) 4.38 (dd,
J = 10.45, 2.65 Hz, 1H) 4.22 (dd, J = 10.29, 5.93 Hz,
1H) 3.86 (m, 1H) 3.19 (d, J = 7.18 Hz, 2H). MS (ESI):
m/z (M+H)+ 467.
5.2.52.
(S)-2-[5-(1H-Indazol-6-yl)-pyridin-3-yloxy]-1-
(1H-indol-3-ylmethyl)-ethylamine (63). Stille reaction
with 6-bromoindazole and stannyl material 8 (as de-
scribed for 33) followed by deprotection of the Boc
1
group with TFA (as described for 59) yielded 63. H
NMR (500 MHz, DMSO-d6) d ppm 13.28 (br s, 1H)
10.97 (s, 1H) 8.53 (s, 1H) 8.31 (s, 1H) 8.11 (s, 1H)
7.81–7.86 (m, 3H) 7.65 (s, 2H) 7.58 (d, J = 7.5 Hz, 1H)
7.41 (d, J = 7.5 Hz, 1H) 7.34 (d, J = 7.5 Hz, 1H) 7.24
(s, 1H), 6.95–7.05 (m, 2H) 4.13 (m, 2H) 3.60 (m, 1H)
2.97 (m, 2H). MS (ESI): m/z (M+H)+ 384.
5.2.49. (S)-1-(1H-Indol-3-ylmethyl)-2-{5-[3-(1H-pyrrol-2-
yl)-1H-indazol-5-yl]-pyridin-3-yloxy}-ethylamine
5.2.53. (S)-2-{5-[3-(1H-Imidazol-2-yl)-1H-indazol-5-yl]-
pyridin-3-yloxy}-1-(1H-indol-3-ylmethyl)-ethylamine (58).
Stille reaction with 5-bromo-3-(imidazol-2-yl)indazole
(26 R = imidazol-2-yl) and stannyl material 8 (as de-
scribed for 33) followed by deprotection of the Boc group
(57).
Stille reaction with 5-bromo-3-(pyrol-2-yl)indazole (31)
and stannyl material 8 (as described for 33) followed
by deprotection of the Boc group with TFA (as de-
scribed for 59) yielded 57. 1H NMR (300 MHz,
DMSO-d6) d ppm 13.10 (br s, 1H) 11.38 (s, 1H) 11.03
(s, 1H) 8.68 (s, 1H) 8.35 (d, J = 2.37 Hz, 1H) 8.26 (s,
1H) 8.15 (br s, 2H) 7.67 (m, 4H) 7.38 (d, J = 8.14 Hz,
1H) 7.30 (d, J = 2.03 Hz, 1H) 7.10 (t, J = 7.46 Hz, 1H)
7.01 (t, J = 7.46 Hz, 1H) 6.86 (m, 2H) 6.21 (m, 1H)
4.38 (m, 1H) 4.20 (dd, J = 10.51, 5.76 Hz, 1H) 3.87 (m,
1H) 3.18 (m, 2H). MS (ESI) m/z (M+H)+ 449. Anal.
Calcd for C27H24N6OÆ2.5TFA: C, 52.39; H, 3.64; N,
11.46. Found: C, 52.26; H, 3.67; N, 11.39.
1
with TFA (as described for 59) yielded 58. H NMR
(500 MHz, DMSO-d6) d ppm 14.36 (s, 1H) 11.04 (s, 1H)
8.72 (s, 1H) 8.61 (s, 1H) 8.39 (d, J = 2.50 Hz, 1H) 8.26
(br s, 3H) 7.86 (s, 2H) 7.83 (s, 1H) 7.76 (s, 1H) 7.63 (d,
J = 8.11 Hz, 1H) 7.38 (d, J = 8.11 Hz, 1H) 7.30 (d,
J = 2.18 Hz, 1H) 7.09 (t, J = 7.49 Hz, 1H) 7.00 (t,
J = 7.49 Hz, 1H) 4.37 (dd, J = 10.45, 2.96 Hz, 1H) 4.22
(dd, J = 10.45, 5.77 Hz, 1H) 3.86 (s, 1H) 3.19 (d,
J = 7.17 Hz, 2H). MS (ESI): m/z (M+H)+ 450.
5.2.54. (S)-2-[5-(1H-Benzotriazol-5-yl)-pyridin-3-yloxy]-
1-(1H-indol-3-ylmethyl)-ethylamine (65). Stille reaction
with N-Boc-5-bromobenztriazole (36) and stannyl mate-
rial 8 (as described for 33) followed by deprotection of
5.2.50. (S)-(5-{5-[2-Amino-3-(1H-indol-3-yl)-propoxy]-
pyridin-3-yl}-1H-indazol-3-yl)-dimethyl-amine (60). Stille
reaction with 5-bromo-3-(N,N-dimethylamino)indazole

K. W. Woods et al. / Bioorg. Med. Chem. 14 (2006) 6832–6846
6845
1
the Boc group with TFA (as described for 59) yielded 65.
1H NMR (300 MHz, DMSO-d6) d ppm 11.03 (s, 1H)
8.66 (d, J = 1.36 Hz, 1H) 8.38 (d, J = 2.71 Hz, 1H)
8.27 (m, 1H) 8.18 (m, 3H) 8.01 (m, 1H) 7.78 (m, 1H)
7.63 (d, J = 7.80 Hz, 1H) 7.38 (d, J = 8.14 Hz, 1H)
7.30 (d, J = 2.37 Hz, 1H) 7.10 (t, J = 7.12 Hz, 1H) 7.01
(t, J = 6.95 Hz, 1H) 4.38 (dd, J = 10.68, 2.88 Hz, 1H)
4.21 (dd, J = 10.68, 6.27 Hz, 1H) 3.86 (m, 1H) 3.17 (d,
J = 7.12 Hz, 2H). MS (ESI): m/z (M+H)+ 385.
trifluoroacetate salt. H NMR (500 MHz, DMSO-d6) d
ppm 13.64 (m, 1H) 11.03 (s, 1H) 8.59 (s, 1H) 8.37 (s, 1H)
8.32 (s, 1H) 8.26 (s, 3H) 7.77 (m, 2H) 7.71 (s, 1H) 7.63 (d,
J = 7.80 Hz, 1H) 7.38 (d, J = 8.11 Hz, 1H) 7.30
(d, J = 1.87 Hz, 1H) 7.10 (t, J = 7.33 Hz, 1H) 7.01 (t,
J = 7.33 Hz, 1H) 4.37 (dd, J = 10.29, 2.50 Hz, 1H) 4.21
(dd, J = 10.29, 5.93 Hz, 1H) 3.86 (m, 1H) 3.18 (d,
J = 7.49 Hz, 2H). MS (ESI): m/z (M+H)+ 428.
5.2.57.
(S)-6-{5-[2-Amino-3-(1H-indol-3-yl)-propoxy]-
5.2.55. (S)-2-[5-(3-Benzyl-1H-indazol-5-yl)-pyridin-3-yloxy]
-1-(1H-indol-3-ylmethyl)-ethylamine (54). Stille reaction
with 5-bromo-3-benzylindazole (26 R = benzyl) and
stannyl material 8 (as described for 33) followed by
deprotection of the Boc group with TFA (as described
pyridin-3-yl}-1H-indazol-3-ylamine (64). The product
was prepared as described for compound 59 using 4-bro-
mo-2-fluorobenzonitrile in place of the 5-bromo-2-fluoro-
1
benzonitrile. H NMR (300 MHz, DMSO-d6) d 11.93
(br s, 1H) 11.03 (s, 1H) 8.60 (d, J = 1.36 Hz, 1H) 8.37
(d, J = 2.37 Hz, 1H) 8.17 (s, 4H) 7.88 (d, J = 8.14 Hz,
1H) 7.71 (s, 1H) 7.63 (m, 2H) 7.34 (m, 3H) 7.07 (m,
2H) 4.35 (m, 1H) 4.19 (s, 1H) 3.91 (d, J = 30.85 Hz,
1H) 3.16 (d, J = 5.42 Hz, 2H). MS (ESI): m/z (M+H)+
399. Anal. Calcd for C23H22N6OÆ3.5TFA: C, 45.18;H,
3.22; N, 10.54, F, 25.01. Found: C, 44.83; H, 3.19; N,
10.40, F, 25.01.
1
for 59) yielded 54. H NMR (300 MHz, DMSO-d6) d
ppm 11.03 (s, 1H) 8.55 (d, J = 1.70 Hz, 1H) 8.32 (d,
J = 2.71 Hz, 1H) 8.15 (m, 3H) 7.98 (s, 1H) 7.61 (m,
4H) 7.29 (m, 7H) 7.08 (m, 1H) 7.02 (t, J = 7.12 Hz,
1H) 4.42 (dd, J = 10.68, 5.93 Hz, 1H) 4.35 (s, 2H) 4.17
(dd, J = 10.68, 5.93 Hz, 1H) 3.86 (m, 1H) 3.17 (m,
2H). MS (ESI): m/z (M+H)+ 474. Anal. Calcd for
C30H27N5OÆ3.9TFA: C, 49.44; H, 3.39; N, 7.63. Found:
C, 49.07; H, 3.75; N, 7.42.
5.3. Biology/testing
5.2.56. 5-{5-[(S)-2-Amino-3-(1H-indol-3-yl)-propoxy]-
pyridin-3-yl}-1H-indazole-3-carboxylic acid (62)
Details of the biological assays and testing protocols
have been reported previously.22,36
5.2.56.1. Step 1. 1H-Indazole-3-carboxylic acid methyl
ester. A solution of 3-carboxyindazole (2.0 g; 12.3 mmol)
and concd HCl (2 mL) in MeOH (50 mL) was heated at
reflux overnight. The reaction mixture was concentrated,
diluted with 2 N NaOH (aq), and extracted with EtOAc.
The extracts were rinsed with brine, dried over MgSO4,
and concentrated to provide methyl indazole-3-carbox-
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6846
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