Discovery of selective glucocorticoid receptor modulator MK-5932

Article abstract of DOI:10.1016/j.bmc.2011.10.054

A series of partial agonists of the Glucocorticoid Receptor were prepared targeting reduced transactivation activity, while maintaining significant transrepression activity. Incorporation of an ortho-aryl amide produced compounds with the desired in vitro

Full text of DOI:10.1016/j.bmc.2011.10.054

Bioorganic & Medicinal Chemistry 19 (2011) 7374–7386
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Bioorganic & Medicinal Chemistry
journal homepage: www.elsevier.com/locate/bmc
Discovery of selective glucocorticoid receptor modulator MK-5932
Christopher J. Bungard a, , George D. Hartman a, Jesse J. Manikowski a, James J. Perkins a, Chang Bai b,
⇑
Philip E. Brandish b, Danielle H. Euler b, James C. Hershey b, Azriel Schmidt b, Yulin Fang c,
Ryan T. Norcross c, Tom H. Rushmore c, Charles D. Thompson c, Robert S. Meissner a
a Department of Medicinal Chemistry, Merck Research Laboratories, 770 Sumneytown Pike, West Point, PA 19486, USA
b Department of Molecular Endocrinology, Merck Research Laboratories, 770 Sumneytown Pike, West Point, PA 19486, USA
c Department of Drug Metabolism Merck Research Laboratories, 770 Sumneytown Pike, West Point, PA 19486, USA
a r t i c l e i n f o
a b s t r a c t
Article history:
A series of partial agonists of the Glucocorticoid Receptor were prepared targeting reduced transactiva-
tion activity, while maintaining significant transrepression activity. Incorporation of an ortho-aryl amide
produced compounds with the desired in vitro profile. Bioreactors consisting of Suspension cultures of
Sf21 cells co expressing a CYP3A4 and NADPH-cytochrome P450 oxireductase were used to prepare
the major metabolites of these compounds and revealed that oxidative N-dealkylation provided a path-
way for formation of metabolites that were more agonistic than the parent partial agonists. Oxidative
N-dealkylation was blocked in a new series of compounds, however oxidation alone was capable of pro-
ducing full agonist metabolites. Incorporation of an ortho-primary amide and utilization of fluorine to
modulate agonism afforded partial agonist MK-5932. Synthesis of the major metabolites of MK-5932
using bioreactor technology revealed that no significant GR-active metabolites were formed. Orally
administered MK-5932 displayed anti-inflammatory efficacy in a Rat Oxazolone-induced chronic derma-
titis model, while sparing plasma insulin.
Received 16 August 2011
Revised 10 October 2011
Accepted 18 October 2011
Available online 24 October 2011
Keywords:
Glucocorticoids
Inflammation
Selective glucocorticoid receptor modulator
SEGRM
Ó 2011 Elsevier Ltd. All rights reserved.
1. Introduction
(SEGRM’s). A SEGRM is a molecule designed to activate only a sub-
set of the functions induced by the endogenous ligand in hope of
Glucocorticoids are some of the most commonly prescribed
drugs for the treatment of inflammatory and immunological disor-
ders such as asthma1 and rheumatoid arthritis.2 Unfortunately,
chronic use of these therapeutic agents carries significant risks in
the form of the accompanying serious, sometimes irreversible side
effects.3 These include, but are not limited to, osteoporosis,4
diabetes,5 glaucoma,6 and muscle atrophy.7,8 Efforts to discover
novel ways to minimize these undesirable side effects while main-
taining the desired anti-inflammatory activity are ongoing. One of
the earliest advances toward this goal was the design of new
ligands that offer improved selectivity for the glucocorticoid recep-
tor (GR) over other nuclear hormone receptors such as the Miner-
alocorticoid Receptor (MR), however GR mediated side effects still
remained a problem.9 Topical10 and inhaled11 delivery of Glucocor-
ticoids has provided a means for reducing their systemic side
effects, thus increasing the therapeutic window for drugs adminis-
tered by this route. However, not all diseases are amenable to top-
ical or inhaled routes of administration. A novel approach to safely
treating inflammatory diseases with Glucocorticoids would be the
use of so-termed selective glucocorticoid receptor modulators
retaining the desired anti-inflammatory benefits, while reducing
the undesirable side effects.12
In order to formulate an appropriate target in vitro profile for a
SEGRM, the current state of glucocorticoid (GC) biology needs to be
considered. The GR resides in the cytoplasm in an inactive form
associated with various heat shock proteins and other chaper-
ones.13 GR ligands, after passing through the plasma membrane,
bind to the GR resulting in formation of an activated ligand/recep-
tor complex. This activated complex dissociates from some, but not
all, of the associated proteins and translocates into the cell nu-
cleus.14 In its homodimeric form,15 the activated GR-ligand com-
plex can positively regulate target genes by binding to specific
DNA binding sites, referred to as positive glucocorticoid responsive
elements (GRE’s).16 This process, termed transactivation, results in
the synthesis of both anti-inflammatory proteins, such as Lipocor-
tin-117, along with regulator proteins, such as tyrosine aminotrans-
ferase (TAT)18 and phosphoenolpyruvate carboxykinase (PEPCK)19
which are enzymes involved in gluconeogenesis. Transactivation
is thought to play a minor role in eliciting the desired anti-inflam-
matory action of Glucocorticoids and a more important role in pro-
ducing the numerous side effects of these drugs. The negative
regulation of target genes by the GR is more complicated and can
occur through multiple mechanisms. For example, the activated
⇑
Corresponding author. Tel.: +1 215 362 3409; fax: +1 215 652 7310.
E-mail address: christopher_bungard@merck.com (C.J. Bungard).
0968-0896/$ - see front matter Ó 2011 Elsevier Ltd. All rights reserved.
doi:10.1016/j.bmc.2011.10.054

C. J. Bungard et al. / Bioorg. Med. Chem. 19 (2011) 7374–7386
7375
Table 1
GR-ligand complex can bind to negative GRE’s and possibly inter-
Compounds described in Scheme 2
fere with the binding of transcription factors, leading to repression
of transcription, as has been documented for the regulation of oste-
ocalcin.20,21 Alternatively, the activated GR-ligand monomer can
interact directly (i.e., protein–protein interaction) with transcrip-
Compound
R1
R2
R3
R4
R5
7a, 8a
7b, 8b
7c, 8c
7d, 8d
7e, 8e
7f, 8f
9a
9b
9c
9d
9e
CO2Me
H
H
CO2Me
CO2Me
CO2Me
CO2H
H
H
CO2Et
H
F
H
H
H
CO2H
H
H
H
H
H
H
H
H
F
H
H
H
H
H
F
H
H
H
H
H
H
H
H
H
H
H
H
tion factors such as AP-122 or NF- B23 that are involved in regulat-
j
CO2Me
H
F
H
H
H
CO2H
H
F
ing the expression of pro-inflammatory cytokines and
inflammatory mediators such as interleukin-1 (IL-1) and tumor
necrosis factor-a (TNF-a). This pathway, termed transrepression,
does not require binding of the GR to DNA and is thought to play
a major role in producing the desired anti-inflammatory activity
of Glucocorticoids. The concept that transactivation and transre-
pression are mechanistically separable has been probed by muta-
tion of the GR in the DNA binding domain. This prevents
transactivation, without altering transrepression of AP-1.24 Accord-
ingly, compounds that exhibit reduced transactivation activity,
while maintaining significant transrepression activity, may retain
the desired anti-inflammatory activity of current glucocorticoids,
with a reduced propensity to induce one or more of the undesir-
able side effects associated with these compounds.25 Efforts to-
ward developing such SEGRM’s are ongoing, and some promising
in vitro and in vivo data has been reported, however clinical
proof-of-concept in patients has yet to be achieved.26
Our efforts in this area were inspired by reports describing the
discovery of C,D-ring truncated analogues of Fluorocortivazol as
non-steroidal dissociating GR ligands, shown in Figure 1. It was
hypothesized that the steroidal C- and D-rings could be effectively
mimicked by incorporation of an appropriate aryl group such as in
2, or arylethyl group such as in 3.27,28 We began our efforts with an
analysis of an overlay of compounds such as 3 with the crystal
structure of dexamethasone in the human GR ligand binding do-
main.29 This revealed that although a simple arylethyl group is
an effective mimic of the hydrophobic component of the steroidal
C,D-rings, key hydrogen bonding interactions, between the polar
functional groups on the D-ring of dexamethasone and residues
Q642 and T739, were not being made by compounds such as 3.
Thus, the starting point for our Medicinal Chemistry efforts was
to incorporate a substituent capable of hydrogen bonding on the
right hand side aryl ring of 3 to potentially rediscover the hydrogen
bonding interactions which were lost during the truncation of 1 to
afford 3.
H
CO2H
CO2H
CO2H
F
H
H
9f
H
10a
H
H
H
H
H
N
H
O
10b
10c
H
H
H
H
H
H
H
H
10d
10e
10f
10g
10h
10i
H
H
H
H
H
H
H
H
H
H
H
H
H
H
H
H
H
H
H
H
H
N
H
O
O
H
H
F
N
H
N
H
O
O
N
H
F
N
H
O
O
H
H
N
H
O
O
10j
N
H
10k
10l
H
F
H
H
H
H
H
H
H
H
H
N
H
H2N
O
10m
H
N
H
O
O
2. Chemistry
10n
10o
10p
F
H
H
F
H
H
H
H
H
H
N
H
Synthesis of our target molecules was designed to enable late
stage incorporation of substituted phenethyl substituents via a
Sonagashira/Hydrogenation protocol. Synthesis of the Sonagashira
precursor was carried out as detailed in Scheme 1. Addition of lith-
ium TMS-acetylide to the Hajos-Parrish Ketone30 afforded tertiary
alcohol 4. Low temperature formylation of 4, followed by silyl
deprotection of the acetylene produced the desired b-ketoaldehyde
5. Treatment of crude 5 with p-fluororphenylhydrazine hydrochlo-
ride and NaOAc in acetic acid afforded the desired indazole acety-
lene 6.
HO
HO
F
N
H
O
O
H
N
H
10q
10r
H
H
H
H
F
H
F
H2N
H2N
O
O
H
OAc
O
HO
Ar
H
OH
Ar
OH
HO
C
D
N
N
N
N
N
A
B
N
CH3
Fluorocortivazol 1
F
F
2
3
F
Figure 1. Fluorocortivazol 1 and truncated analogues 2 and 3.

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C. J. Bungard et al. / Bioorg. Med. Chem. 19 (2011) 7374–7386
OH
OH
OH
O
TMS
N
OHC
O
a
b
c
N
O
O
4
5
6
F
Scheme 1. Reagents and conditions: (a) nBuLi, TMS-acetylene, THF, ꢀ78 °C, 80% (b) (i) LDA, ethyl formate, THF,ꢀ78 °C; (ii) K2CO3, MeOH, rt, 75%; (c) p-fluorophenylhydr-
azineꢁHCl, NaOAc, HOAc, rt, 94%.
R4
R4
R1
R5
R3
R2
R5
R3
R2
HO
OH
HO
R1
b
a
N
N
N
N
N
N
6
7
8
F
F
F
R4
R1
R4
R5
R3
R2
R5
R3
R2
HO
HO
N
N
c
d
R1
N
N
9
10
F
F
Scheme 2. Reagents and conditions: (a) 2% (PPh3)2PdCl2, 2% CuI, ArX, iPr2NH, THF, 80 °C; (b) 10% Pd/C, H2 balloon, rt; (c) 1 M NaOH, EtOH, 100 °C; (d) R3NH2, HATU, iPr2NEt,
DMF, rt.
With the ultimate goal of incorporating a small alkyl amide as
the hydrogen bonding substituent, our general synthetic approach
is outlined in Scheme 2. The appropriate aryl halides were coupled
with 6 to afford acetylenes 7a–q. Subsequent hydrogenation affor-
ded the desired phenethyl esters 8a–q, which were readily hydro-
lyzed to afford acids 9a–q. The desired targets were obtained via
HATU coupling of acids 9a–q with appropriate amines to afford
amides 10a–q (Table 1).31
and Glucocorticoid regulation and modulation of MMTV assay to
afford a measure of transrepression and transactivation activity,
respectively. It is evident from the data in Table 2 for compounds
10a–c that the preferred position for a hydrogen bonding substitu-
ent is the ortho position with respect to the phenethyl linker.
10d R1
10e R1
10f R1
10g R1
10h R1
10i R1
=
=
=
=
=
=
R2 = H
R2 = H
R2 = H
R2 = F
R2 = F
R2 = H
N
H
O
O
O
O
3. Results
N
H
Compounds 10a–c, shown in Figure 2, were evaluated for GR
activity in a binding assay and transcriptional activity was evalu-
OH
N
H
R2
ated using the glucocorticoid induced TNF-
a repression assay
N
R1
N
R3
N
H
OH
10a R2, R3 = H, R1
10b R1, R3 = H, R2
10c R1, R2 = H, R3
=
=
=
O
O
O
N
H
R2
F
N
R1
N
H
N
N
H
O
O
N
H
N
H
F
Figure 2. Regioisomeric arylamides 10a–c.
Figure 3. Arylamides 10d–i.
Table 2
In vitro data for regioisomeric aryl amides
Compound
GRBind32
Ki (nM)
TNF-
a
repression33
Emax (%dex)
MMTV (A549)34
MMTV (HeLa)35
IP (nM)
IP (nM)
Emax (%dex)
IP (nM)
Emax (%dex)
10a
10b
10c
2
15
155
263
nd
nd
50
<10
12
193
nd
nd
49
<10
<10
317
nd
nd
48
<10
<10

C. J. Bungard et al. / Bioorg. Med. Chem. 19 (2011) 7374–7386
7377
Table 3
Binding and transcriptional data for ortho-arylamides 10d–i
Compound
GRBind32
Ki (nM)
TNF-
a
repression33
Emax (%dex)
MMTV (A549)34
Emax (%dex)
MMTV (HeLa)35
Emax (%dex)
IP (nM)
IP (nM)
IP (nM)
10d
10e
10f
10g
10h
10i
1
2
4
<1
<1
2
123
127
170
119
30
62
50
62
67
72
65
62
81
60
95
26
39
37
29
48
24
25
51
93
85
83
132
43
107
21
17
30
16
17
38
88
OH
OH
OH
N
N
CYP3A4
O
NH
O
NH
N
N
10i
11
F
F
CYP3A4
OH
OH
N
N
O
HO
NH
N
O
NH2
N
10j
F
F
Figure 4. Major metabolites of 10i.
Table 4
In vitro data for 10i and metabolites 10j and 11
Compound
GRBind32
Ki (nM)
TNF-
a
repression33
Emax (%dex)
MMTV (A549)34
MMTV (HeLa)35
IP (nM)
IP (nM)
Emax (%dex)
IP (nM)
Emax (%dex)
10i
10j
11
2
2
40
88
56
958
65
82
34
39
61
1564
51
60
20
107
130
1325
38
66
6
10k R1
=
=
=
R2 = H
N
H
O
O
O
O
OH
10l R1
10m R1
10n R1
R2 = F
R2
N
H
N
R1
N
HO
HO
R2 = H
R2 = F
N
H
F
=
N
H
Figure 5. Arylamides 10k–n.
Table 5
Compounds incapable of oxidative N-dealkylation
Compound
GRBind32
Ki (nM)
TNF-
a
repression33
Emax (%dex)
MMTV (A549)34
Emax (%dex)
MMTV (HeLa)35
IP (nM)
IP (nM)
IP (nM)
Emax (%dex)
10k
10l
10m
10n
10
<1
1
246
20
26
79
81
88
96
212
32
33
23
41
83
108
27
30
14
33
71
79
3
24
23
104
35

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C. J. Bungard et al. / Bioorg. Med. Chem. 19 (2011) 7374–7386
R4
R5
R3
R2
10o R2 = F, R3-5 = H
10p R3 = F, R2,4,5 = H
10q R4 = F, R2,3,5 = H
10r R5 = F, R2-4 = H
OH
N
O
NH2
N
F
Figure 6. Arylamides 10o–r.
Encouraged by this result we prepared the series of compounds,
shown in Figure 3, containing an ortho-arylamide substituent.
Pleasingly, a number of compounds were generated that displayed
reduced activity in the transactivation assay, while maintaining
potency and activity in the transrepression assay as shown in Table
3.
Figure 7. In vivo activity of 10p in the Rat Oxazolone-induced chronic dermatitis
model.
Of particular concern moving forward was the potential for
compounds to undergo metabolic transformations in an in vivo
setting and produce metabolites that displayed GR activity. As
we sought partial agonists of the GR, active metabolites that be-
have as antagonists could block the desired anti-inflammatory
activity of the parent molecule. On the contrary, metabolites that
behave as full agonists could erode any potential separation of
anti-inflammatory activity from adverse effects that might be char-
acteristic of the parent partial agonist. Accordingly, we sought a
method to generate sufficient quantities of metabolites to enable
structural elucidation in addition to testing for GR activity in our
in vitro assays. The synthesis of milligram quantities of P450-gen-
erated metabolites has been accomplished by employing so termed
‘bioreactors’ consisting of a suspension culture of Sf21 insect cells,
co-infected with baculovirus containing the cDNA for a single cyto-
chrome P450, and NADPH-cytochrome P450 oxireductase.36 We
chose to evaluate the metabolic profile of compound 10i in a
CYP3A4 bioreactor system. The metabolic profile of 10i is outlined
in Figure 4. Cytochrome P450 mediated hydroxylation at the 4-po-
sition affords 11, a compound with much weaker binding affinity
than the parent and very little transcriptional activity. However,
oxidative N-dealkylation of the aryl amide moiety afforded the pri-
mary amide 10j, a compound that exhibits a higher level of agon-
ism than the parent. Thus, oxidative N-dealkylation provides a
mechanism for generation of metabolites more agonistic than the
parent molecule, and renders compounds capable of this transfor-
mation unsuitable for further development. This was exemplified
when 10i was dosed at 20 mpk in a Rat Oxazolone-induced chronic
dermatitis model.37 Compound 10i achieved exposure of 75
however GR active compound 10j was also present at an exposure
of 50 M h thereby confounding interpretation of the in vivo
lM h,
l
result. Consistent in vivo results could not be obtained with com-
pound 10i possibly due to the presence of metabolite 10j. Metab-
olite 10j was independently tested in the Rat Oxazolone-induced
chronic dermatitis model and at doses of 5 and 25 mpk and
Figure 8. LC–MS traces from in vitro metabolic profiling of 10m in the rat and
human liver microsomes.
achieved exposures of 134 and 199 lM h, respectively. While 10j
Table 6
Modulation of agonism by fluorine incorporation
Compound
GRBind32
TNF-
a
repression33
Emax (%dex)
MMTV (A549)34
Emax (%dex)
MMTV (HeLa)35
Emax (%dex)
IP (nM)
IP (nM)
IP (nM)
10j
2
<1
3
<1
<1
56
10
131
89
3
82
84
73
70
90
61
9
186
36
6
60
75
35
120
82
130
11
153
58
7
66
58
23
47
89
10o
10p
10q
10r

C. J. Bungard et al. / Bioorg. Med. Chem. 19 (2011) 7374–7386
7379
F
OH
OH
N
N
O
O
NH2
N
N
M1
a
F
OH
F
F
F
N
OH
O
NH2
b
N
10p
NH2
OH
M3
M2
F
CH3
F
F
N
OH
c
N
N
O
HO
O
HO
O
N
F
F
Scheme 3. Reagents and conditions: (a) CYP3A4 bioreactor; (b) (i) SeO2, dioxane, 50 °C, 65%; (ii) Dess–Martin, CH2Cl2, rt, 94%; (iii) NaBH4, EtOH, 0 °C, 41%. (c) DDQ, HOAc, rt,
quant.
Table 7
In vitro profiles of key metabolites of MK-5932
Compound
GRBind32
TNF-
a
repression33
Emax (%dex)
MMTV (A549)34
Emax (%dex)
MMTV (HeLa)35
Emax (%dex)
IP (nM)
IP (nM)
IP (nM)
MK-5932
M1
M2
3
58
>400
>400
131
nd
nd
73
6
<5
<5
186
nd
nd
35
<5
<5
<5
153
nd
nd
23
<5
<5
<5
M3
nd
nd
nd
displayed significant anti-inflammatory activity, the compound
also produced significant effects on plasma insulin at both doses,
confirming its ability to interfere with our ability to interpret the
in vivo effects of 10i (Table 4).
In order to minimize the risk of active metabolite generation
a series of compounds incapable of undergoing the undesirable
oxidative N-dealkylation were prepared as shown in Figure 5.
As shown in Table 5, compounds 10k and 10l possessed an
in vitro profile which we envisioned would afford separation of
anti-inflammatory activity from adverse effects in an in vivo
Compound 10p was evaluated in the Rat Oxazolone-induced
chronic dermatitis model37 as depicted below in Figure 7. Follow-
ing oral administration (90:10 PEG400/H2O) it was shown that 10p
was capable of producing an anti-inflammatory response similar to
that produced by 6MP. Interestingly, it was shown that although
retaining significant anti-inflammatory activity, 10p produced no
dose responsive effects on plasma insulin. This is in direct contrast
to 6MP which significantly elevated plasma insulin levels.
Although encouraged by this result we wanted to examine the
potential of 10p to produce active metabolites as this could signif-
icantly affect the compound’s ability to retain the observed SEGRM
activity when moving across species. In order to evaluate this
potential, liver microsome incubations were carried out on radiola-
belled 10p, the results of which are presented in Figure 8.
One major metabolite M1, and a minor metabolite M2, were
present in both rat and human microsome incubations. Of particu-
lar concern was metabolite M3 which was present in human but
not rat incubations. Thus it was important for us to establish the
identity and in vitro profile of M1, M2, and particularly of M3
whose GR activity may not have been captured in our rat in vivo
model. The identity of M1, M2, and M3 were established by a
combination of human P450 bioreactor incubations and chemical
synthesis as outlined in Scheme 3.
setting. When compound 10l was evaluated at
15 mpk in the Rat Oxazolone-induced chronic dermatitis model
an exposure of 64 M h was achieved. However, much to our
dismay full agonist metabolite 10n was also detected at an
a dose of
l
exposure of 3.8
to interpret.
lM h making the overall in vivo readout difficult
Guided by the above results it seemed that it was not plausible
to have a functional group in the ortho-position of the aryl moiety
that was capable of undergoing metabolic oxidation. With this in
mind we sought a compound containing a primary amide in the
ortho-position that exhibited the desired dissociation profile. It
was reasoned that introduction of a fluorine into the appropriate
position of the RHS aryl ring of 10j may perturb the receptor en-
ough to afford a compound of the desired profile. Accordingly,
compounds 10k–n in Figure 6 were synthesized and gratifyingly
compound 10p afforded a promising in vitro profile, as shown in
Table 6, that prompted us to further evaluate it in an in vivo
setting.
Metabolite M1 was isolated from a CYP3A4 bioreactor incuba-
tion, and was shown to be a C4-hydroxylated species, similar to
what was observed for 10i. Metabolites M2 and M3 were prepared
by chemical synthesis. The in vitro data for M1, M2, and M3 is
shown in Table 7.

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C. J. Bungard et al. / Bioorg. Med. Chem. 19 (2011) 7374–7386
Pleasingly none of the observed in vitro metabolites displayed
5.1.2. (3R,3aS)-3-Ethynyl-3-hydroxy-3a-methyl-6-oxo-
any significant activity in the GR transcriptional assays. In addition
to this, aside from metabolites M1, M2, and M3, no significant lev-
els of additional circulating metabolites were observed in vivo in
rats. These observations pave the way for evaluation of 10p, now
designated MK-5932, in advanced studies to validate the SEGRM
hypothesis. Further results pertaining to the Biological activity of
MK-5932 will be reported in due course.
2,3,3a,4,5,6-hexahydro-1H-indene-5-carbaldehyde (5)
A 1.5 M solution of lithium diisopropylamide mono(tetrahydro-
furan) in cyclohexane (121 mL, 182 mmol) was added to a solution
of 4 (9.54 g, 36.4 mmol) in THF (400 mL) at ꢀ78 °C and the result-
ing solution stirred at this temperature for 1 h to afford a thick sus-
pension. Methyl formate (22.6 mL, 364 mmol) was added dropwise
over about 15 min and the resulting suspension stirred at ꢀ78 °C
for 5 h. The reaction was quenched at ꢀ78 °C with 1 M aqueous
HCl solution and the aqueous layer checked to ensure it was acidic.
The crude product was extracted with EtOAc (ꢂ3) and the com-
bined organic extracts were dried over anhydrous MgSO4 and the
solvent removed in vacuo to afford crude the crude product, MS
(ESI): m/z = 291.18 (MH+), 78% pure by LC–MS, that was used di-
rectly in the next step without purification. Removal of the TMS-
group was accomplished by adding K2CO3 (5.03 g, 72.8 mmol) to
a solution of the crude material in MeOH (300 mL), and stirring
the resulting suspension stirred at ambient temperature for
90 min. The MeOH was removed in vacuo and 1 M aqueous HCl
was added to the residue and the crude product extracted with
EtOAc (ꢂ3). The combined organic extracts were dried over anhy-
drous MgSO4 and the solvent removed in vacuo. Purification by
flash chromatography on 330 g of silica, eluting with a gradient
of 0–70% EtOAc in hexanes afforded 5.94 g, 75% of 5 as a tan solid.
1H NMR (CDCl3, 500 MHz): d 7.38 (s, 1H), 5.89 (t, J = 1.95 Hz, 1H),
3.15 (d, J = 13.9 Hz, 1H), 2.69 (s, 1H), 2.58–2.75 (m, 2H), 2.20–
2.33 (m, 3H), 1.87 (s, OH), 1.14 (s, 3H). MS (ESI): m/z = 219.25
(MH+), 100% pure by LC–MS.
4. Conclusion
A series of ortho-amide containing compounds were prepared
that offered promising in vitro profiles. Bioreactor guided optimi-
zation of the structures to minimize formation of active metabo-
lites led to incorporation of a primary amide as metabolically
tolerable functionality. Incorporation of fluorine to modulate agon-
ism produced MK-5932, an orally efficacious SEGRM capable of
producing significant anti-inflammatory activity, while producing
no effects on plasma insulin levels in the Rat Oxazolone-induced
chronic dermatitis model.
5. Experimental
5.1. Chemistry
All commercially available reagents and solvents were used
without further purification unless otherwise stated. Automated
flash chromatography was performed on an Isco CombiFlashÒ
Companion™ using RedisepÒ disposable flash cartridges with
peak detection at 254 nm. Reverse phase purification was accom-
plished using a Gilson 215 liquid handler equipped with Gemini
5.1.3. (4aS,5R)-5-Ethynyl-1-(4-fluorophenyl)-4a-methyl-
1,4,4a,5,6,7-hexahydrocyclopenta[f]indazol-5-ol (6)
NaOAc (41.3 g, 504 mmol) was added to a solution of 5 (100 g,
458 mmol) and 4-fluorophenylhydrazine hydrochloride (1–5)
(82 g, 504 mmol) in acetic acid (916 mL) and the resulting suspen-
sion stirred at ambient temperature for 1 h. The reaction was
quenched slowly (caution CO2 evolution) with saturated aqueous
NaHCO3 solution and the crude product extracted with EtOAc
(ꢂ3). The combined organic extracts were dried over anhydrous
MgSO4 and the solvent removed in vacuo. Purification by flash
chromatography on 1.5 kg of silica, eluting with a gradient of 0–
100% EtOAc in hexanes afforded 133 g, 94% of 6 as a tan solid. 1H
NMR (CDCl3, 500 MHz): d 7.45 (m, 2H), 7.43 (s, 1H), 7.15 (M, 2H),
6.18 (m, 1H), 3.35 (d, J = 15.4 Hz, 1H), 2.70 (s, 1H), 2.62 (d,
J = 15.6 Hz, 1H), 2.54–2.62 (m, 2H), 2.23–2.32 (m, 2H), 1.73 (s,
OH), 1.14 (s, 3H). MS (ESI): m/z = 309.2 (MH+), 100% pure by LC–
MS.
C18 column (20 ꢂ 100 mm or 30 ꢂ 100 mm I.D., 5
lm) with a
0.1% TFA in water/acetonitrile gradient. Peak collection was trig-
gered by UV detection at 215 or 254 nm. 1H NMR spectra were
recorded on
a Unity Inova 500 instrument operating at
500 MHz with tetramethylsilane or residual protiated solvent
used as a reference. Analytical LC was performed on a Waters
2695 separations module (YMC Pro C18 column, 50 ꢂ 3 mm
I.D., S-5 lM, 12 nm particle size); 0.05% TFA in water/acetonitrile
gradient; UV detection @ 215 and 254 nm). Low resolution mass
spectra were recorded using a Waters Micromass ZQ with elec-
trospray ionization. High resolution mass spectra were recorded
using electrospray ionization (ESI) on a 7T Fourier transform
ion cyclotron resonance (FTICR) mass spectrometer. Unless other-
wise noted, all reactions were conducted in flame dried
glassware.
5.1.4. General procedure for the synthesis of acetylenes 7a–g
Diisopropylamine (1.0 equiv) was added to a solution of 6
(1.0 equiv), aryl halide (1.2 equiv), bis(triphenylphosphine)palla-
dium(II) chloride (2 mol %), and CuI (2 mol %) in anhydrous THF
(0.25 M) at ambient temperature. The resulting solution was stir-
red at 80 °C for 1 h, then diluted with diethyl ether, filtered
through a pad of celite, and the solvent removed in vacuo. Purifica-
tion by flash chromatography on silica, eluting with a gradient of
0–100% EtOAc in hexanes afforded the desired products.
5.1.1. (1S,7aS)-1-Hydroxy-7a-methyl-1-[(trimethylsilyl)ethynyl]
-1,2,3,6,7,7a-hexahydro-5H-inden-5-one (4)
A 2.5 M solution of nBuLi (27.4 mL, 68.5 mmol) in hexanes
was added dropwise to a solution of trimethylsilylacetylene
(9.48 mL, 68.5 mmol) in THF (90 mL) at ꢀ78 °C. The resulting
solution was stirred at ꢀ78 °C for 30 min, then a solution of Ha-
jos-Parrish Ketone (1–1, 7.5 g, 45.7 mmol) in THF (90 mL) was
added and the resulting solution stirred at ꢀ78 °C for 30 min.
The reaction was quenched with saturated aqueous KH2PO4
and the crude product extracted with EtOAc (ꢂ3). The combined
organic extracts were dried over anhydrous MgSO4 and the sol-
vent removed in vacuo. Purification by flash chromatography on
120 g of silica, eluting with a gradient of 0–55% EtOAc in hex-
anes afforded 9.54 g, 80% of 4 as a white solid. 1H NMR (CDCl3,
500 MHz): d 5.80 (t, J = 2.0 Hz, 1H), 2.70 (ddd, J = 19.5, 9.0,
2.2 Hz, 1H), 2.40–2.60 (m, 4H), 2.24 (m, 2H), 1.85 (m, 1H),
1.21 (s, 3H), 0.11 (s, 9H). MS (ESI): m/z = 263.25 (MH+), 100%
pure by LC–MS.
5.1.4.1. Methyl 2-{[(4aS,5R)-1-(4-fluorophenyl)-5-hydroxy-4a-
methyl-1,4,4a,5,6,7-hexahydrocyclopenta[f]indazol-5-yl]ethy-
nyl}benzoate (7a).
91% Yield; 1H NMR (CDCl3, 500 MHz): d
7.91 (dd, J = 7.8, 1.2 Hz, 1H), 7.43 (s, 1H), 7.41–7.49 (m, 5H), 7.35
(td, J = 7.8, 1.5 Hz, 2H), 6.20 (t, J = 2.0 Hz, 1H), 3.84 (s, 3H), 3.43
(d, J = 15.6 Hz, 1H), 2.77 (s, OH), 2.72 (d, J = 15.4 Hz, 1H),
2.65–2.69 (m, 2H), 2.05–2.45 (m, 2H), 1.21 (s, 3H); MS (ESI):
m/z = 443.14 (MH+), 100% pure by LC–MS.

C. J. Bungard et al. / Bioorg. Med. Chem. 19 (2011) 7374–7386
7381
5.1.4.2. Ethyl 3-{[(4aS,5R)-1-(4-fluorophenyl)-5-hydroxy-4a-
methyl-1,4,4a,5,6,7-hexahydrocyclopenta[f]indazol-5-yl]ethy-
nyl}benzoate (7b).
5.1.5.2. Ethyl 3-{2-[(4aS,5S)-1-(4-fluorophenyl)-5-hydroxy-4a-
methyl-1,4,4a,5,6,7-hexahydrocyclopenta[f]indazol-5-
yl]ethyl}benzoate (8b).
91% Yield; 1H NMR (CDCl3, 500 MHz): d
91% Yield; 1H NMR (CDCl3,
8.05 (m, 1H), 7.97 (m, 1H), 7.56 (dt, J = 7.8, 1.5 Hz, 1H), 7.45 (s,
1H), 7.45–7.48 (m, 2H), 7.37 (t, J = 7.8 Hz, 1H), 7.15 (m, 2H), 6.21
(m, 1H), 4.37 (q, J = 7.1 z, 2H), 3.41 (d, J = 15.4 Hz, 1H), 2.71 (d,
J = 15.4 Hz, 1H), 2.62–2.68 (m, 2H), 2.32–2.44 (m, 2H), 2.28 (s,
OH), 1.39 (t, J = 7.1 Hz, 3H), 1.19 (s, 3H); MS (ESI): m/z = 457.15
(MH+), 100% pure by LC–MS.
500 MHz): d 7.88 (s, 1H), 7.86 (d, J = 7.6 Hz, 1H), 7.43–7.47
(m, 2H), 7.40 (m, 2H), 7.34 (t, J = 7.6 Hz, 1H), 7.14 (m, 2H),
6.19 (m, 1H), 4.37 (q, J = 7.1 Hz, 1H), 2.89–2.95 (m, 1H), 2.88
(d, J = 15.1 Hz, 1H), 2.76–2.82 (m, 1H), 2.59–2.65 (m, 1H), 2.45
(d, J = 15.1 Hz, 1H), 2.32–2.40 (m, 1H), 2.16–2.21 (m, 1H), 1.94–
2.01 (m, 1H), 1.84–1.90 (m, 1H), 1.39 (q, J = 7.1 Hz, 3H), 1.16
(s, 3H); MS (ESI): m/z = 461.20 (MH+), 96% pure by LC–MS.
5.1.4.3. Methyl 4-{[(4aS,5R)-1-(4-fluorophenyl)-5-hydroxy-4a-
methyl-1,4,4a,5,6,7-hexahydrocyclopenta[f]indazol-5-yl]ethy-
nyl}benzoate (7c).
7.95 (m, 2H), 7.45 (s, 1H), 7.43–7.48 (m, 4H), 7.15 (t, J = 8.5 Hz,
2H), 6.21 (m, 1H), 3.91 (s, 3H), 3.39 (d, J = 15.4 Hz, 1H), 2.71 (d,
J = 15.4 Hz, 1H), 2.58–2.70 (m, 2H), 2.35–2.42 (m, 2H), 2.33 (s,
OH), 1.19 (s, 3H); MS (ESI): m/z = 443.14 (MH+), 100% pure by
LC–MS.
5.1.5.3. Methyl 4-{2-[(4aS,5S)-1-(4-fluorophenyl)-5-hydroxy-4a-
methyl-1,4,4a,5,6,7-hexahydrocyclopenta[f]indazol-5-
yl]ethyl}benzoate (8c).
500 MHz): d 7.95 (d, J = 8.1 Hz, 2H), 7.43–7.46 (m, 2H), 7.40 (s,
1H), 7.26–7.28 (m, 2H), 7.13–7.16 (m, 2H), 6.19 (m, 1H), 3.90 (s,
3H), 2.89–2.96 (m, 1H), 2.88 (d, J = 15.4 Hz, 1H), 2.76–2.82 (m,
1H), 2.59–2.65 (m, 1H), 2.46 (d, J = 15.1 Hz, 1H), 2.30–2.38 (m,
1H), 2.15–2.20 (m, 1H), 1.94–2.01 (m, 1H), 1.84–1.90 (m, 1H),
1.72–1.78 (m, 1H), 1.16 (s, 3H); MS (ESI): m/z = 447.19 (MH+),
100% pure by LC–MS.
98% Yield; 1H NMR (CDCl3, 500 MHz): d
92% Yield; 1H NMR (CDCl3,
5.1.4.4.
hydroxy-4a-methyl-1,4,4a,5,6,7-hexahydrocyclopenta[f]inda-
zol-5-yl]ethynyl}benzoate (7d).
90% Yield; 1H NMR (CDCl3,
Methyl
2-fluoro-6-{[(4aS,5R)-1-(4-fluorophenyl)-5-
500 MHz): d 7.44–7.46 (m, 1H), 7.43 (s, 1H), 7.31–7.36 (m, 1H),
7.33 (m, 1H), 7.24 (dd, J = 7.8, 0.8 Hz, 1H), 7.15 (m, 2H), 7.08 (td,
J = 8.8, 1.0 Hz, 1H), 6.20 (t, J = 2.0 Hz, 1H), 3.80 (s, 3H), 3.33 (d,
J = 15.4 Hz, 1H), 2.69 (d, J = 15.4 Hz, 1H), 2.61–2.66 (m, 2H), 2.55
(s, OH), 2.34–2.37 (m, 2H), 1.18 (s, 3H); MS (ESI): m/z = 461.15
(MH+); 100% pure by LC–MS.
5.1.5.4. Methyl 2-fluoro-6-{2-[(4aS,5R)-1-(4-fluorophenyl)-5-
hydroxy-4a-methyl-1,4,4a,5,6,7-hexahydrocyclopenta[f]inda-
zol-5-yl]ethyl}benzoate (8d).
92% Yield; 1H NMR (CDCl3,
500 MHz): d 7.43–7.47 (m, 2H), 7.38 (s, 1H), 7.30–7.34 (m, 1H),
7.12–7.16 (, 2H), 7.03 (d, J = 7.3 Hz, 1H), 6.96 (m, 1H), 6.18 (m,
1H), 3.92 (s, 3H), 2.90–2.96 (m, 1H), 2.82 (d, J = 15.1 Hz, 1H),
2.75–2.80 (m, 1H), 2.58–2.63 (m, 1H), 2.43 (d, J = 15.1 Hz, 1H),
2.32–2.40 (m, 1H), 2.09 ddd, J = 12.9, 8.8, 2.2 Hz, 1H), 1.96 (s,
OH), 1.94–2.01 (m, 1H), 1.80–1.86 (m, 1H), 1.72–1.78 (m,
1H), 1.16 (s, 3H); MS (ESI): m/z = 465.17 (MH+); 93% pure by LC–
MS.
5.1.4.5.
hydroxy-4a-methyl-1,4,4a,5,6,7-hexahydrocyclopenta[f]inda-
zol-5-yl]ethynyl}benzoate (7e).
100% Yield; 1H NMR (CDCl3,
Methyl
5-fluoro-2-{[(4aS,5R)-1-(4-fluorophenyl)-5-
500 MHz): d 7.61 (m, 1H), 7.42–7.47 (m, 4H), 7.11–7.16 (m, 3H),
6.19 (m, 1H), 3.84 (s, 3H), 3.41 (d, J = 15.4 Hz, 1H), 2.71 (d,
J = 15.4 Hz, 1H), 2.64–2.68 (m, 2H), 2.35–2.42 (, 2H), 1.20 (s, 3H);
MS (ESI): m/z = 461.16 (MH+); 100% pure by LC–MS.
5.1.5.5. Methyl 5-fluoro-2-{2-[(4aS,5R)-1-(4-fluorophenyl)-5-
hydroxy-4a-methyl-1,4,4a,5,6,7-hexahydrocyclopenta[f]inda-
zol-5-yl]ethyl}benzoate (8e).
95% Yield; 1H NMR (CDCl3,
5.1.4.6.
hydroxy-4a-methyl-1,4,4a,5,6,7-hexahydrocyclopenta[f]inda-
zol-5-yl]ethynyl}benzoate (7f).
92% Yield; 1H NMR (CDCl3,
Methyl
4-fluoro-2-{[(4aS,5R)-1-(4-fluorophenyl)-5-
500 MHz): d 7.63 (dd, J = 9.8, 2.9 Hz, 1H), 7.42–7.45 (m, 2H), 7.36
(s, 1H), 7.22 (dd, J = 8.5, 5.6 Hz, 1H), 7.12–7.15 (m, 3H), 6.17 (m,
1H), 3.90 (s, 3H), 3.19–3.24 (m, 1H), 2.90–2.96 (m, 1H), 2.78 (d,
J = 15.4 Hz, 1H), 2.58–2.64 (m, 1H), 2.43 (d, J = 15.1 Hz, 1H), 2.36–
2.42 (m, 1H), 2.14–2.19 (m, 1H), 2.07 (s, OH), 1.99–2.06 (m, 1H),
1.70–1.80 (m, 2H), 1.81 (s, 3H); MS (ESI): m/z = 465.19 (MH+);
96% pure by LC–MS.
500 MHz): d 7.95 (dd, J = 8.8, 5.9 Hz, 1H), 7.46 (dd, J = 9.0, 4.9 z,
2H), 7.43 (s, 1H), 7.13–7.26 (m, 3H), 7.04 (dd, J = 8.3, 2.7 Hz, 1H),
6.21 (m, 1H), 3.84 (s, 3H), 3.39 (d, J = 15.4 Hz, 1H), 2.82 (s, OH),
2.72 (d, J = 15.4 Hz, 1H), 2.65–2.68 (m, 2H), 2.41–2.42 (m,
2H), 1.20 (s, 3H); MS (ESI): m/z = 461.16 (MH+); 100% pure by
LC–MS.
5.1.5.6. Methyl 4-fluoro-2-{2-[(4aS,5R)-1-(4-fluorophenyl)-5-
hydroxy-4a-methyl-1,4,4a,5,6,7-hexahydrocyclopenta[f]inda-
zol-5-yl]ethyl}benzoate (8f).
500 MHz): d 7.98 (m, 1H), 7.42–7.45 (m, 2H), 7.36 (s, 3H), 7.12–
7.16 (m, 2H), 6.92–6.97 (m, 2H), 6.17 (m, 1H), 3.88 (s, 3H), 3.25–
3.31 (m, 1H), 2.90–2.96 (m, 1H), 2.78 (d, J = 15.4 Hz, 1H), 2.72 (s,
OH), 2.58–2.64 (m, 1H), 2.43 (d, J = 15.4 Hz, 1H), 2.36–2.42 (m,
1H), 2.15 (dd, J = 8.6, 2.0 Hz, 1H), 2.00–2.07 (m, 1H), 1.74–1.83
(m, 2H), 1.18 (s, 3H); MS (ESI): m/z = 465.20 (MH+); 91% pure by
LC–MS.
5.1.5. General procedure for the synthesis of compounds 8a–g
10% Pd/C (0.4 equiv of Pd) was added to a solution of 7a–g
(1 equiv) in EtOAc (128 mL) at ambient temperature and the flask
evacuated and backfilled with hydrogen. The resulting suspension
was stirred at ambient temperature under a balloon of hydrogen
for the indicated time, filtered through a pad of celite and the sol-
vent removed in vacuo to afford the desired products, used without
further purification.
92% Yield; 1H NMR (CDCl3,
5.1.5.1. Methyl 2-{2-[(4aS,5R)-1-(4-fluorophenyl)-5-hydroxy-4a-
methyl-1,4,4a,5,6,7-hexahydrocyclopenta[f]indazol-5-
yl]ethyl}benzoate (8a).
500 MHz): d 7.95 (d, J = 8.1 Hz, 2H), 7.43–7.46 (m, 2H), 7.40 (s,
1H), 7.26–7.28 (m, 2H), 6.19 (m, 1H), 3.90 (s, 3H), 2.89–2.96 (m,
1H), 2.88 (d, J = 15.4 Hz, 1H), 2.76–2.82 (m, 1H), 2.59–2.65 (m,
1H), 2.46 (d, J = 15.1 Hz, 1H), 2.30–2.38 (m, 1H), 2.15–2.20 (m,
1H), 1.94–2.00 (m, 1H), 1.84–1.90 (m, 1H), 1.72–1.78 (m, 1H),
1.16 (s, 3H); MS (ESI): m/z = 447.16 (MH+), 100% pure by LC–MS.
5.1.6. General procedure for the synthesis of acids 9a–g
A 1 M aqueous solution of NaOH (2.0 equiv) was added to a
solution of 8a–g (1.0 equiv) in EtOH (0.25 M) and the resulting sus-
pension heated at 100 oC for 1 h. The ethanol was removed in va-
cuo and 1 M aqueous HCl solution was added and the product
was extracted with EtOAc (ꢂ3). The combined organic extracts
were dried over anhydrous MgSO4 and the solvent removed in va-
cuo to afford the desired products.
99% Yield; 1H NMR (CDCl3,

7382
C. J. Bungard et al. / Bioorg. Med. Chem. 19 (2011) 7374–7386
5.1.6.1.
2-{2-[(4aS,5R)-1-(4-Fluorophenyl)-5-hydroxy-4a-
5.1.7. General procedure for the synthesis of amides 10a–r
PYBOPÒ TPYBOP (1 equiv) was added to a solution of the appro-
priate amine (1.5 equiv), Hünig’s Base (3.0 equiv) and the appropri-
ate acid (1.0 equiv) in anhydrous DMF (0.20 M) at ambient
temperature. The resulting solution was stirred at ambient temper-
ature for 1 h. The DMF was removed in vacuo, water was added
and the crude product was extracted with EtOAc (ꢂ3). The com-
bined organic extracts were washed with saturated sodium bicar-
bonate solution, dried over anhydrous MgSO4, filtered and the
solvent removed in vacuo. Purification by flash chromatography
on silica, eluting with a gradient of 0–100% CHCl3 to 70:20:10
CHCl3/EtOAc/MeOH afforded the following products.
methyl-1,4,4a,5,6,7-hexahydrocyclopenta[f]indazol-5-
yl]ethyl}benzoic acid (9a).
98% Yield; 1H NMR (CD3OD,
500 MHz): d 7.86 (d, J = 7.8 Hz, 1H), 7.42–7.46 (m, 3H), 7.37 (s,
1H), 7.32 (d, J = 7.8 Hz, 1H), 7.23–7.26 (m, 3H), 6.14 (m, 1H),
3.31–3.34 (m, 1H), 2.95 (dd, J = 12.0, 4.6 Hz, 1H), 2.82 (d,
J = 15.1 Hz, 1H), 2.48–2.61 (m, 2H), 2.45 (d, J = 15.4 Hz, 1H), 2.21–
2.25 (m, 1H), 1.98–2.05 (m, 1H), 1.70–1.80 (m, 2H), 1.13 (s, 3H);
MS (ESI): m/z = 433.05 (MH+), 100% pure by LC–MS.
5.1.6.2.
3-{2-[(4aS,5S)-1-(4-Fluorophenyl)-5-hydroxy-4a-
methyl-1,4,4a,5,6,7-hexahydrocyclopenta[f]indazol-5-
yl]ethyl}benzoic acid (9b).
97% Yield; 1H NMR (CD3OD,
5.1.7.1.
methyl-1,4,4a,5,6,7-hexahydrocyclopenta[f]indazol-5-yl]ethyl}-
N-methylbenzamide (10a).
49% Yield; 1H NMR (CDCl3,
2-{2-[(4aS,5R)-1-(4-Fluorophenyl)-5-hydroxy-4a-
500 MHz): d 7.98 (m, 1H), 7.83(dt, J = 7.8, 1.5 Hz, 1H), 7.44–
7.47 (m, 2H), 7.39 (s, 1H), 7.37 (t, J = 7.6 Hz, 2H), 7.23–7.27 (m,
2H), 6.18 (m, 1H), 2.91 (td, J = 12.5, 4.2 Hz, 1H), 2.86 (d,
J = 15.6 Hz, 1H), 2.76–2.82 (m, 1H), 2.58–2.63 (m, 1H), 2.48 (d,
J = 15.4 Hz, 1H), 2.35–2.44 (m, 1H), 2.13–2.17 (m, 1H),
2.00–2.07 (m, 1H), 1.81–1.86 (m, 1H), 1.69–1.75 (m, 1H),
1.13 (s, 3H); MS (ESI): m/z = 433.20 (MH+), 97% pure by
LC–MS.
500 MHz): d 7.42–7.45 (m, 2H), 7.32–7.36 (m, 2H), 7.35 (s, 1H),
7.20–7.24 (m, 2H), 7.12–7.15 (m, 2H), 6.15 (m, 1H), 5.97 (m, NH),
3.03–3.08 (m, 1H), 2.99 (d, J = 4.9 Hz, 3H), 2.80 (d, J = 15.4 Hz,
1H), 2.71–2.79 (m, 1H), 2.55–2.61 (m, 1H), 2.44 (d, J = 15.4 Hz,
1H), 2.31–2.39 (m, 1H), 2.00–2.08 (m, 2H), 1.91–1.97 (m, 1H),
1.77–1.84 (m, 1H), 1.16 (s, 3H); HRMS: calcd for C27H29FN3O2
(MH)+ 446.2238, found 446.2237; 100% pure by LC–MS.
5.1.6.3. 4-{2-[(4aS,5S)-1-(4-fluorophenyl)-5-hydroxy-4a-methyl-
1,4,4a,5,6,7-hexahydrocyclopenta[f]indazol-5-yl]ethyl}benzoic
acid (9c).
J = 8.1 Hz, 2H), 7.44–7.47 (m, 2H), 7.39 (s, 1H), 7.33 (d, J = 8.1 Hz,
2H), 7.23–7.27 (m, 2H), 6.18 (m, 1H), 2.89–2.95 (m, 1H), 2.85 (d,
J = 15.6 Hz, 1H), 2.77–2.82 (m, 1H0, 2.58–2.63 (m, 1H), 2.48 (d,
J = 15.4 Hz, 1H), 2.35–2.42 (m, 1H), 2.12–2.16 (m, 1H), 2.02–2.07
(m, 1H), 1.81–1.86 (m, 1H), 1.69–1.75 (m, 1H), 1.14 (s, 3H); MS
(ESI): m/z = 433.18 (MH+), 97% pure by LC–MS.
5.1.7.2.
methyl-1,4,4a,5,6,7-hexahydrocyclopenta[f]indazol-5-yl]ethyl}-
N-methylbenzamide (10b).
94% Yield; 1H NMR (CDCl3,
3-{2-[(4aS,5S)-1-(4-Fluorophenyl)-5-hydroxy-4a-
67% Yield; 1H NMR (CD3OD, 500 MHz): d 7.92 (d,
500 MHz): d 7.65 (s, 1H), 7.51–7.53 (m, 1H), 7.43–7.46 (m, 2H),
7.39 (s, 1H), 7.32–7.33 (m, 2H), 7.14 (t, J = 8.5 Hz, 2H), 6.17 (m,
1H), 6.16 (m, NH), 3.72 (m, 1H), 3.18 (dd, J = 14.9, 7.3 Hz, 1H),
3.00 (d, J = 4.9 Hz, 3H), 2.88–2.94 (m, 1H), 2.87 (d, J = 15.4 Hz,
1H), 2.75–2.80 (m, 1H), 2.58–2.64 (m, 1H), 2.44 (d, J = 15.4 Hz,
1H), 2.31–2.38 (m, 1H), 2.15–2.20 (m, 1H), 1.94–2.01 (m, 1H),
1.84–1.89 (m, 1H), 1.72–1.78 (m, 1H), 1.16 (s, 3H); HRMS: calcd
for C27H29FN3O2 (MH)+ 446.2238, found 446.2248; 91% pure by
LC–MS.
5.1.6.4. 2-Fluoro-6-{2-[(4aS,5R)-1-(4-fluorophenyl)-5-hydroxy-
4a-methyl-1,4,4a,5,6,7-hexahydrocyclopenta[f]indazol-5-
yl]ethyl}benzoic acid (9d).
100% Yield; 1H NMR (CDCl3,
500 MHz): d 7.38–7.46 (m, 3H), 7.30 (dd, J = 13.7, 7.8 Hz, 1H),
7.13 (t, J = 8.3 Hz, 2H), 7.00 (d, J = 7.6 Hz, 1H), 6.93–6.97 (m, 1H),
6.10 (s, 1H), 3.00–3.05 (m, 1H), 2.79 (d, J = 15.4 Hz, 1H), 2.75–
2.80 (, 1H), 2.58 (dd, J = 18.6, 9.3 Hz, 1H), 2.45 (d, J = 15.4 Hz, 1H),
2.31–2.39 (m, 1H), 1.99–2.12 (m, 2H), 1.89–1.94 (m, 1H), 1.74–
1.81 (m, 1H); 1.15 (s, 3H); MS (ESI): m/z = 451.14 (MH+); 94% pure
by LC–MS.
5.1.7.3.
methyl-1,4,4a,5,6,7-hexahydrocyclopenta[f]indazol-5-yl]ethyl}-
N-methylbenzamide (10c).
87% Yield; 1H NMR (CDCl3,
4-{2-[(4aS,5S)-1-(4-Fluorophenyl)-5-hydroxy-4a-
500 MHz): d 7.67 (d, J = 8.1 Hz, 2H), 7.43–7.45 (m, 2H), 7.40 (s,
1H), 7.24–7.26 (m, 1H), 7.13–7.16 (m, 2H), 6.19 (m, 1H), 6.12 (m,
NH), 3.00 (d, J = 4.9 Hz, 3H), 2.89–2.94 (m, 1H), 2.87 (d,
J = 15.4 Hz, 1H), 2.86–2.94 (m, 1H), 2.74–2.80 (m, 1H), 2.58–2.64
(m, 1H), 2.46 (d, J = 15.4 Hz, 1H), 2.29–2.37 (m, 1H), 2.14–2.19
(m, 1H), 1.94–2.01 (m, 1H), 1.83–1.89 (m, 1H), 1.71–1.77 (m,
1H), 1.16 (s, 3H); HRMS: calcd for C27H29FN3O2 (MH)+ 446.2238,
found 446.2245; 100% pure by LC–MS.
5.1.6.5. 5-Fluoro-2-{2-[(4aS,5R)-1-(4-fluorophenyl)-5-hydroxy-
4a-methyl-1,4,4a,5,6,7-hexahydrocyclopenta[f]indazol-5-
yl]ethyl}benzoic acid (9e).
100% Yield; 1H NMR (CDCl3,
500 MHz): d 7.72 (dd, J = 9.3, 2.7 Hz, 1H), 7.43–7.46 (m, 2H), 7.41
(s, 1H), 7.21 (dd, J = 8.6, 5.6 Hz, 1H), 7.12–7.16 (m, 3H), 6.15 (m,
1H), 3.30–3.35 (m, 1H), 2.78–2.85 (m, 1H), 2.79 (d, J = 15.6 Hz,
1H), 2.61 (dd, J = 18.8, 9.8 Hz, 1H), 2.47 (d, J = 15.4 Hz, 1H), 2.33–
2.41 (m, 1H), 2.14–2.18 (m, 1H), 2.04–2.11 (m, 1H), 1.78–1.85
(m, 2H), 1.20 (s, 3H); MS (ESI): m/z = 451.18 (MH+); 94% pure by
LC–MS.
5.1.7.4.
4a-methyl-1,4,4a,5,6,7-hexahydrocyclopenta[f]indazol-5-
yl]ethyl}benzamide (10d).
48% Yield; 1H NMR (CDCl3,
N-Allyl-2-{2-[(4aS,5R)-1-(4-Fluorophenyl)-5-hydroxy-
500 MHz): d 7.42–7.46 (m, 2H), 7.39 (d, J = 7.8 Hz, 1H), 7.35 (s,
1H), 7.34–7.37 (m, 1H), 7.21–7.26 (m, 2H), 7.12–7.15 (m, 2H),
6.15 (m, 1H), 6.00 (m, NH), 5.89–5.97 (m, 1H), 5.25–5.29 (m, 1H),
5.18–5.20 (m, 1H), 4.02–4.12 (m, 2H), 3.04–3.09 (m, 1H), 2.80 (d,
J = 15.4 Hz, 1H), 2.73–2.78 (m, 1H), 2.55–2.61 (m, 1H), 2.44 (d,
J = 15.4 Hz, 1H), 2.31–2.39 (m, 1H), 1.99–2.08 (m, 2H), 1.90–1.95
(m, 1H), 1.78–1.84 (m, 1H), 1.16 (s, 3H); HRMS: calcd for
5.1.6.6. 4-Fluoro-2-{2-[(4aS,5R)-1-(4-fluorophenyl)-5-hydroxy-
4a-methyl-1,4,4a,5,6,7-hexahydrocyclopenta[f]indazol-5-
yl]ethyl}benzoic acid (9f).
100% Yield; 1H NMR (CDCl3,
C
29H31FN3O2 (MH)+ 472.2395, found 472.2404; 100% pure by LC–
500 MHz): d 8.06–8.09 (m, 1H), 7.43–7.46 (m, 2H), 7.41 (s, 1H),
7.15 (t, J = 8.5 hz, 1H), 6.94–6.98 (m, 2H), 6.16 (s, 1H), 3.38–3.43
(m, 1H), 2.80–2.85 (m, 1H), 2.80 (d, J= 15.1 Hz, 1H), 2.59–2.64
(m, 1H), 2.48 (d, J = 15.4 Hz, 1H), 2.34–2.42 (m, 1H), 2.05–2.17
(m, 2H), 1.81–1.90 (m, 2H), 1.21 (s, 3H); MS (ESI): m/z = 451.18
(MH+); 89% pure by LC–MS.
MS.
5.1.7.5.
2-{2-[(4aS,5R)-1-(4-Fluorophenyl)-5-hydroxy-4a-
methyl-1,4,4a,5,6,7-hexahydrocyclopenta[f]indazol-5-yl]ethyl}-
N-propylbenzamide (10e).
92% Yield; 1H NMR (CDCl3,
500 MHz): d 7.42–7.45 (m, 2H), 7.32–7.36 (m, 2H), 7.35 (s, 1H),

C. J. Bungard et al. / Bioorg. Med. Chem. 19 (2011) 7374–7386
7383
7.19–7.23 (m, 2H), 7.13 (t, J = 8.5 Hz, 2H), 6.15 (m, 1H), 5.96 (m,
NH), 3.38–3.41 (m, 3H), 3.03–3.08 (m, 1H), 2.80 (d, J = 15.4 Hz,
1H), 2.55–2.60 (m, 1H), 2.44 (d, J = 15.4 Hz, 1H), 2.30–2.38 (m,
1H), 1.99–2.06 (m, 2H), 1.90–1.97 (m, 1H), 1.78–1.84 (m, 1H),
1.59–1.66 (m, 3H), 1.16 (s, 3H), 0.98 (t, J = 7.3 Hz, 3H); HRMS: calcd
for C29H33FN3O2 (MH)+ 474.2551, found 474.2548; 100% pure by
LC–MS.
2.60 (m, 1H), 2.46 (d, J = 15.1 Hz, 1H), 2.41–2.49 (m, 1H), 2.13–
2.17 (m, 1H), 1.97–2.04 (m, 1H), 1.73–1.84 (m, 1H), 1.12 (s, 3H);
HRMS: calcd for C26H27FN3O2 (MH)+ 432.2082, found 432.2083;
100% pure by LC–MS.
5.1.7.11.
hydroxy-4a-methyl-1,4,4a,5,6,7-hexahydrocyclopenta[f]inda-
zol-5-yl]ethyl}benzamide (10k).
97% Yield; 1H NMR (CDCl3,
N-(tert-butyl)-2-{2-[(4aS,5R)-1-(4-Fluorophenyl)-5-
5.1.7.6.
methyl-1,4,4a,5,6,7-hexahydrocyclopenta[f]indazol-5-yl]ethyl}-
N-isopropylbenzamide (10f).
76% Yield; 1H NMR (CDCl3,
2-{2-[(4aS,5R)-1-(4-Fluorophenyl)-5-hydroxy-4a-
500 MHz): d 7.43–7.45 (m, 2H), 7.35 (s, 1H0, 7.30–7.33 (m, 2H),
7.21 (d, J = 8.1 Hz, 2H), 7.12–7.15 (m, 2H), 6.15 (s, 1H), 5.73 (s,
NH), 3.02–3.07 (m, 1H), 2.84 (d, J = 15.4 Hz, 1H), 2.70–2.76 (m,
1H), 2.55–2.60 (m, 1H), 2.45 (d, J = 15.4 Hz, 1H), 2.31–2.39 (m,
1H), 2.00–2.03 (m, 2H), 1.90–1.94 (m, 1H), 1.79–1.85 (m, 1H),
1.47 (s, 9H), 1.17 (s, 3H); HRMS: calcd for C30H35FN3O2 (MH)+
488.2708, found 488.2701; 100% pure by LC–MS.
500 MHz): d 7.42–7.45 (m, 2H), 7.35 (s, 1H), 7.32–7.35 (m, 2H),
7.20–7.23 (m, 2H), 7.12–7.15 (m, 2H), 6.15 (m, 1H), 5.73
(d, J = 7.8 Hz, NH), 4.23–4.30 (m, 1H), 3.04–3.09 (m, 1H), 2.81 (d,
J = 15.4 Hz, 1H), 2.71–2.77 (m, 1H), 2.55–2.61 (m, 1H), 2.44 (d,
J = 15.4 Hz, 1H), 2.31–2.39 (m, 1H), 1.98–2.07 (m, 2H), 1.90–1.95
(m, 1H), 1.78–1.84 (m, 1H), 1.27 (d, J = 6.3 Hz, 3H), 1.25 (d,
J = 6.6 Hz, 3H), 1.16 (s, 3H); HRMS: calcd for C29H33FN3O2 (MH)+
474.2551, found 474.2565; 100% pure by LC–MS.
5.1.7.12.
methyl-1,4,4a,5,6,7-hexahydrocyclopenta[f]indazol-5-yl]ethyl}-
N-(1-methylcyclopropyl)benzamide (10l).
63% Yield; 1H
2-{2-[(4aS,5R)-1-(4-Fluorophenyl)-5-hydroxy-4a-
NMR (CDCl3, 500 MHz): d 7.42–7.45 (m, 2H), 7.36 (s, 1H), 7.23–
7.27 (m, 1H), 7.12–7.15 (m, 2H), 7.00 (d, J = 7.6 Hz, 1H), 6.91 (t,
J = 8.5 Hz, 1H), 6.24 (s, 1H), 6.16 (m, 1H), 2.91–2.97 (m, 1H), 2.81
(d, J = 15.4 Hz, 1H), 2.68–2.74 (m, 1H), 2.55–2.61 (m, 1H), 2.45 (d,
J = 15.1 Hz, 1H), 2.31–2.39 (m, 1H), 1.96–2.05 (m, 2H), 1.89–1.96
(m, 1H), 1.76–1.82 (m, 1H), 1.51 (s, 3H), 1.16 (s, 3H), 0.85–0.91
(m, 2H), 0.71–0.77 (m, 2H); HRMS: calcd for C30H32F2N3O2 (MH)+
504.2457, found 504.2460; 100% pure by LC–MS.
5.1.7.7. 2-Fluoro-6-{2-[(4aS,5R)-1-(4-fluorophenyl)-5-hydroxy-
4a-methyl-1,4,4a,5,6,7-hexahydrocyclopenta[f]indazol-5-
yl]ethyl}-N-(2-methylprop-2-en-1-yl)benzamide
(10g).
69% Yield; 1H NMR (CDCl3, 500 MHz): d 7.42–7.45 (m,
2H), 7.36 (s, 1H), 7.28–7.31 (m, 1H), 7.12–7.15 (m, 2H), 7.03 (d,
J = 7.8 Hz, 1H), 6.94–6.97 (m, 1H), 6.15 (s, 1H), 6.06 (m, NH), 4.96
(s, 1H), 4.09 (s, 1H), 4.10 (dd, J = 15.6, 6.3 Hz, 1H), 3.96 (dd,
J = 15.9, 5.6 Hz, 1H), 2.95–3.00 (m, 1H), 2.83 (s, OH), 2.80 (d,
J = 15.4 Hz, 1H), 2.73–2.78 (m, 1H), 2.55–2.61 (m, 1H), 2.44 (d,
J = 15.4 Hz, 1H), 2.31–2.38 (m, 1H), 1.98–2.05 (m, 2H), 1.90–1.95
(m, 1H), 1.81 (s, 3H), 1.76–1.84 (m, 1H), 1.57 (s, 3H); HRMS: calcd
for C30H32F2N3O2 (MH)+ 504.2457, found 504.2456; 100% pure by
LC–MS.
5.1.7.13.
methyl-1,4,4a,5,6,7-hexahydrocyclopenta[f]indazol-5-yl]ethyl}-
N-(2-hydroxy-1,1-dimethylethyl)benzamide (10m). 72%
2-{2-[(4aS,5R)-1-(4-Fluorophenyl)-5-hydroxy-4a-
Yield; 1H NMR (CDCl3, 500 MHz): d 7.42–7.45 (m, 2H), 7.36 (s,
1H), 7.32–7.34 (m, 2H), 7.20–7.25 (m, 2H), 7.12–7.15 (, 2H), 6.16
(s, 1H), 5.87 (s, 1H), 3.80 (d, J = 11.5 Hz, 1H), 3.72 (d, J = 11.5 Hz,
1H), 3.09–3.13 (m, 1H), 2.81 (d, J = 15.1 Hz, 1H), 2.71–2.79 (m,
1H), 2.56–2.62 (m, 1H), 2.43 (d, J = 15.4 Hz, 1H), 2.33–2.42 (m,
1H), 2.07–2.11 (m, 1H), 1.81–1.84 (m, 2H), 1.41 (m, 6H), 1.15 (s,
5.1.7.8. 2-Fluoro-6-{2-[(4aS,5R)-1-(4-fluorophenyl)-5-hydroxy-
4a-methyl-1,4,4a,5,6,7-hexahydrocyclopenta[f]indazol-5-
yl]ethyl}-N-propylbenzamide (10h).
66% Yield; 1H NMR
(CDCl3, 500 MHz): d 7.42–7.45 (m, 2H), 7.36 (s, 1H), 7.25–7.29
(m, 1H), 7.12–7.15 (m, 2H), 7.02 (d, J = 7.3 Hz, 1H), 6.94 (m, 1H),
6.15 (m, 1H), 5.98 (m, NH), 3.36–3.50 (m, 2H), 2.94–2.99 (m, 1H),
2.91 (s, OH), 2.81 (d, J = 15.4 Hz, 1H), 2.71–2.79 (m, 1H), 2.55–
2.61 (m, 1H), 2.44 (d, J = 15.4 Hz, 1H), 2.30–2.38 (m, 1H), 1.98–
2.08 (m, 1H), 1.76–1.83 (m, 1H), 1.64 (sextet, J = 7.3 Hz, 2H),
1.23–1.32 (m, 2H), 1.16 (s, 3H), 0.99 (t, J = 7.3 Hz, 3H); HRMS: calcd
for C29H32F2N3O2 (MH)+ 492.2457, found 492.2456; 100% pure by
LC–MS.
3H); HRMS: calcd for
504.2663; 100% pure by LC–MS.
C
30H35FN3O3 (MH)+ 504.2657, found
5.1.7.14.
2-{2-[(4aS,5R)-1-(4-Fluorophenyl)-5-hydroxy-4a-
methyl-1,4,4a,5,6,7-hexahydrocyclopenta[f]indazol-5-yl]ethyl}-
N-[1-(hydroxymethyl)cyclopropyl]benzamide (10n).
76%
Yield; 1H NMR (CDCl3, 500 MHz): d 7.42–7.45 (m, 2H), 7.37 (s,
1H), 7.25–7.29 (m, 1H), 7.14 (t, J = 8.5 Hz, 2H), 7.01 (d, J = 7.6 Hz,
1H), 6.90–6.94 (m, 1H), 6.40 (s, 1H), 6.17 (m, 1H), 3.83 (d,
J = 9.3 Hz, 1H), 3.66–3.68 (m, 2H), 3.11 (td, J = 12.5, 3.2 Hz, 1H),
2.81 (d, J = 15.4 Hz, 1H), 2.72 (td, J = 12.5, 5.9 Hz, 1H), 2.58–2.63
(m, 1H), 2.43 (d, J = 15.4 Hz, 1H), 2.32–2.39 (m, 1H), 2.06–2.11
(m, 1H), 1.95–2.02 (m, 1H), 1.85–1.89 (m, 1H), 1.70–1.77 (m,
1H), 1.15 (s, 3H), 0.94 (m, 4H); HRMS: calcd for C30H32F2N3O3
(MH)+ 520.2406, found 520.2409; 100% pure by LC–MS.
5.1.7.9. N-Ethyl-2-{2-[(4aS,5R)-1-(4-fluorophenyl)-5-hydroxy-
4a-methyl-1,4,4a,5,6,7-hexahydrocyclopenta[f]indazol-5-
yl]ethyl}benzamide (10i).
72% Yield; 1H NMR (CDCl3,
500 MHz): d 7.43–7.45 (m, 2H), 7.35 (s, 1H), 7.32–7.36 (m, 2H),
7.20–7.26 (m, 2H), 7.12–7.15 (m, 2H), 6.15 (s, 1H), 5.90 (m, NH),
3.45–3.51 (m, 2H), 3.39 (s, OH), 3.04–3.09 (m, 1H), 2.81 (d,
J = 15.4 Hz, 1H), 2.71–2.77 (m, 1H), 2.55–2.61 (m, 1H), 2.44 (d,
J = 15.4 Hz, 1H), 2.31–2.38 (m, 1H), 1.97–2.07 (m, 2H), 1.91–1.95
(m, 1H), 1.78–1.84 (m, 1H), 1.25 (t, J = 7.3 Hz, 3H), 1.71 (s, 3H);
HRMS: calcd for C28H31FN3O2 (MH)+ 460.2395, found 460.2395;
100% pure by LC–MS.
5.1.7.15. 2-Fluoro-6-{2-[(4aS,5R)-1-(4-fluorophenyl)-5-hydroxy-
4a-methyl-1,4,4a,5,6,7-hexahydrocyclopenta[f]indazol-5-
yl]ethyl}benzamide (10o).
45% Yield; 1H NMR (CDCl3,
500 MHz): d 7.42–7.47 (m, 2H), 7.37 (s, 1H), 7.29–7.33 (m, 1H),
7.12–7.15 (m, 2H), 7.06 (d, J = 8.1 Hz, 1H), 6.95–6.99 (m, 1H),
6.16 (m, 1H), 6.01 (s, NH), 5.89 (s, NH), 3.02–3.07 (m, 1H), 2.79–
2.85 (m, 1H), 2.81 (d, J = 15.4 Hz, 1H), 2.60 (s, OH), 2.56–2.62 (m,
1H), 2.44 (d, J = 15.4 Hz, 1H), 2.32–2.41 (m, 1H), 2.06–2.11 (m,
1H), 1.95–2.02 (m, 1H), 1.87–1.91 (m, 1H), 1.78–1.84 (m, 1H),
1.16 (s, 3H); HRMS: calcd for C26H26F2N3O2 (MH)+ 450.1988, found
450.1994; 100% pure by LC–MS.
5.1.7.10.
2-{2-[(4aS,5R)-1-(4-Fluorophenyl)-5-hydroxy-4a-
methyl-1,4,4a,5,6,7-hexahydrocyclopenta[f]indazol-5-
yl]ethyl}benzamide (10j).
93% Yield; 1H NMR (CD3OD,
500 MHz): d 7.43–7.46 (m, 2H), 7.37 (s, 1H), 7.34–7.39 (m, 2H),
7.30 (d, J = 7.6 Hz, 1H), 7.20–7.27 (m, 3H), 6.61 (m, 1H), 2.99–
3.07 (m, 1H), 2.88–2.91 (m, 1H), 2.84 (d, J = 15.1 Hz, 1H), 2.55–

7384
C. J. Bungard et al. / Bioorg. Med. Chem. 19 (2011) 7374–7386
5.1.7.16. 5-Fluoro-2-{2-[(4aS,5R)-1-(4-fluorophenyl)-5-hydroxy-
4a-methyl-1,4,4a,5,6,7-hexahydrocyclopenta[f]indazol-5-
yl]ethyl}benzamide (10p).
0.456 mmol), 2-bromo-3-fluorobenzonitrile (91 mg, 0.456 mmol),
1,10-bis(di-tert-butylphosphino)ferrocene palladium dichloride
(30 mg, 0.046 mmol), and Cs2CO3 (446 mg, 1.37 mmol) in degassed
toluene(0.9 mL)/water (0.3 mL) were heated at 80 °C for 1 h. The
reaction was quenched with water and extracted with EtOAc
(ꢂ3). The combined organic extracts were dried over MgSO4, fil-
tered, and concentrated in vacuo. The residue was purified by flash
chromatography on Silica, eluting with a gradient of 0–100% EtOAc
in hexanes to afford 14 (139 mg, 71%) as a white solid; 1H NMR
(CDCl3, 500 MHz): d 7.50 (s, 1H), 7.44–7.40 (m, 3H), 7.33–7.25
(m, 2H), 7.17 (t, J = 8.3 Hz, 2H), 6.15 (s, 1H), 3.18 (td, J = 12.7,
4.5 Hz, 1H), 2.97 (td, J = 12.6, 4.8 Hz, 1H), 2.84 (d, J = 15.4 Hz, 1H),
2.65 (dd, J = 19.0, 9.8 Hz, 1H), 2.53–2.42 (m, 2H), 2.34–2.26 (m,
1H), 2.07–1.98 (m, 1H), 1.88–1.80 (m, 1H), 1.75–1.67 (m, 1H),
1.17 (s, 3H), 0.90–0.82 (m, 1H); MS (ESI): m/z = 432.09 (MH+),
100% pure by LC–MS.
71% Yield; 1H NMR (CDCl3,
500 MHz): d 7.42–7.45 (m, 2H), 7.36 (s, 1H), 7.23 (dd, J = 8.5,
5.6 Hz, 1H), 7.17 (dd, J = 8.8, 2.7 Hz, 1H), 7.12–7.15 (m, 1H), 7.08
(td, J = 8.3, 2.7 Hz, 1H), 6.16 (s, 1H), 5.94 (s, NH), 5.78 (s, NH),
3.94 (s, OH), 3.03–3.09 (m, 1H), 2.81 (d, J = 15.4 Hz, 1H), 2.76–
2.84 (m, 1H), 2.56–2.62 (m, 1H), 2.43 (d, J = 15.4 Hz, 1H), 2.33–
2.41 (m, 1H), 2.06–2.11 (m, 1H), 1.98–2.02 (m, 1H), 1.61–1.88
(m, 2H), 1.15 (s, 3H); HRMS: calcd for C26H26F2N3O2 (MH)+
450.1988, found 450.1998; 100% pure by LC–MS.
5.1.7.17. 4-Fluoro-2-{2-[(4aS,5R)-1-(4-fluorophenyl)-5-hydroxy-
4a-methyl-1,4,4a,5,6,7-hexahydrocyclopenta[f]indazol-5-
yl]ethyl}benzamide (10q).
86% Yield; 1H NMR (CDCl3,
500 MHz): d 7.47 (dd, J = 8.5, 5.7 Hz, 1H), 7.45–7.49 (m, 2H), 7.35
(s, 1H), 7.13 (t, J = 8.4 Hz, 2H), 6.96 (dd, J = 9.6, 2.6 Hz, 1H), 6.91
(td, J = 8.3, 2.6 Hz, 1H), 6.16 (s, 1H), 6.01 (s, 1H), 5.89 (s, 1H),
3.17–3.08 (m, 1H), 2.85–2.78 (m, 2H), 2.59 (dd, J = 18.8, 9.5 Hz,
1H), 2.43 (d, J = 15.3 Hz, 1H), 2.42–2.30 (m, 1H), 2.12–1.94 (m,
2H), 1.91–1.74 (m, 2H), 1.72 (s, 1H), 1.15 (s, 3H); HRMS: calcd
for C26H26F2N3O2 (MH)+ 450.1988, found 450.1985; 100% pure by
LC–MS.
5.1.7.19. 3-Fluoro-2-{2-[(4aS,5R)-1-(4-fluorophenyl)-5-hydroxy-
4a-methyl-1,4,4a,5,6,7-hexahydrocyclopenta[f]indazol-5-
yl]ethyl}benzamide (10r).
25% w/v aq NaOH (52 lL) was
added to a solution of 3-fluoro-2-{2-[(4aS,5R)-1-(4-fluorophenyl)-
5-hydroxy-4a-methyl-1,4,4a,5,6,7-hexahydrocyclopenta[f]indazol-
5-yl]ethyl}benzonitrile (14) (50 mg, 0.12 mmol) in EtOH (1.8 mL)
and the resulting reaction heated at 100 °C in a microwave for
15 min. The reaction was quenched with 1 M HCl and extracted
with EtOAc (ꢂ3). The combined organic extracts were dried over
MgSO4, filtered, and concentrated in vacuo. The residue was puri-
fied by Preparative Reverse Phase HPLC (C-18, 20 ꢂ 150 mm,
Waters Sunfire) Solvent A (0.1% TFA/water), Solvent B (0.1% TFA/
Acetonitrile) 15–85% A/B over 20 min to afford 10r (26 mg, 50%)
as a white solid; 1H NMR (CDCl3, 500 MHz): d 7.56 (s, 1H), 7.43
(dd, J = 8.5, 4.6 Hz, 2H), 7.31–7.13 (m, 5H), 6.45 (s, 1H), 6.20 (s,
1H), 6.09 (s, 1), 3.01 (s, 1H), 2.99–2.89 (m, 1H), 2.83 (d,
J = 15.3 Hz, 1H), 2.62 (dd, J = 19.1, 9.6 Hz, 1H), 2.53–2.34 (m, 2H),
2.15 (t, J = 10.5 Hz, 1H), 2.09–1.97 (m, 1H), 1.86 (t, J = 7.8 Hz, 2H),
1.16 (s, 3H); HRMS: calcd for C26H26F2N3O2 (MH)+ 450.1993, found
450.1993; 100% pure by LC–MS.
5.1.7.18. 3-Fluoro-2-{2-[(4aS,5R)-1-(4-fluorophenyl)-5-hydroxy-
4a-methyl-1,4,4a,5,6,7-hexahydrocyclopenta[f]indazol-5-
yl]ethyl}benzamide (10r) was synthesized from acetylene (6)
according to the following sequence. 5.1.7.18.1. (4aS,5R)-5-
Ethenyl-1-(4-fluorophenyl)-4a-methyl-1,4,4a,5,6,7-hexahydrocyclo-
penta[f]indazol-5-ol (12). (4aS,5R)-5-Ethynyl-1-(4-fluorophenyl)-
4a-methyl-1,4,4a,5,6,7-hexahydrocyclopenta[f]indazol-5-ol
(3.00 g, 9.73 mmol) (6) and Lindlar Catalyst (414 mg) were dis-
solved in THF (65 mL). Pyridine (10 mL) was added and the reac-
tion was evacuated and backfilled with hydrogen then stirred at
ambient temperature under a balloon of hydrogen for 2 h. The
reaction was purged with nitrogen then filtered through Celite
and the solvent removed in vacuo. Purification by flash chromatog-
raphy on Silica, eluting with a gradient of 0–100% EtOAc in hexanes
afforded 12 (2.65 g, 88%) as a white solid; 1H NMR (CDCl3,
500 MHz): d 7.48–7.42 (m, 2H), 7.38 (s, 1H), 7.18–7.11 (m, 2H),
6.19 (s, 1H), 6.03 (dd, J = 17.3, 11.0 Hz, 1H), 5.33 (d, J = 17.3 Hz,
1H), 5.14 (d, J = 11.0 Hz, 1H), 2.83 (d, J = 15.7 Hz, 1H), 2.68 (dd,
J = 19.0, 10.4 Hz, 1H), 2.49–2.38 (m, 2H), 2.26–2.18 (m, 1H), 1.97–
1.91 (m, 1H), 1.66 (s, 1H), 1.19 (s, 3H); MS (ESI): m/z = 311.01
(MH+), 100% pure by LC–MS.
5.2. Biological Evaluation
5.2.1. Competition radioligand binding assay
Human full length GR expressed in insect cells from Invitrogen
(P2812) was incubated with compounds and 3H-dexamethasone at
4 °C overnight before filtration capture of protein and quantitation
of bound radioactivity. An excess of unlabelled dexamethasone
was used to determine non-specific binding. Data is reported as
an average of a minimum of two runs.
5.1.7.18.2. (4aS,5S)-1-(4-Fluorophenyl)-4a-methyl-5-[2-(4,4,5,5-
tetramethyl-1,3,2-dioxaborolan-2-yl)ethyl]-1,4,4a,5,6,7-hexahydrocy-
clopenta[f]indazol-5-ol (13). Pinacolborane (6.40 mL, 44.1 mmol)
was added to
a
solution of (4aS,5R)-5-ethenyl-1-(4-fluoro-
5.2.2. TNF-
U937 cells were stably transfected with a bb-lactamase reporter
construct carrying the human TNF-
promoter (ꢀ1083 to +182).
Assays were carried out in 5% charcoal-stripped FBS. Reporter gene
expression was stimulated by treatment with 1 g/mL LPS and
50 ng/mL TPA. Cells were treated with compounds for 16 h, and
b-lactamase activity was measured using standard methods. Con-
trols were DMSO only and 100 nm dexamethasone. Compound
activity is reported as a percentage of the difference between the
controls. Data is reported as an average of a minimum of three
runs.
a promoter transrepression assay
phenyl)-4a-methyl-1,4,4a,5,6,7-hexahydrocyclopenta[f]indazol-5-
ol (12) (6.85 g, 22.1 mmol), Chloro(1,5-cyclooctadiene)iridium(I)
dimer (222 mg, 0.331 mmol), and Bis(diphenylphosphino)methane
(255 mg, 0.662 mmol) in CH2Cl2 (88 mL) and the reaction heated at
60 °C for 2 h. The solvent was removed in vacuo and the residue
purified by flash chromatography on Silica, eluting with a gradient
of 0–100% EtOAc in hexanes to afford 13 (7.22 g, 75%) as a white
solid; 1H NMR showed a mixture of Boron species, taken forward
as isolated; MS (ESI): m/z = 339.24 (MꢀC6H11O+), 100% pure by
LC–MS.
a
l
5.1.7.18.3. 3-Fluoro-2-{2-[(4aS,5R)-1-(4-fluorophenyl)-5-hydroxy-
4a-methyl-1,4,4a,5,6,7-hexahydrocyclopenta[f]indazol-5-
5.2.3. MMTV promoter transactivation assay
yl]ethyl}benzonitrile
methyl-5-[2-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)ethyl]-
1,4,4a,5,6,7-hexahydrocyclopenta[f]indazol-5-ol (13) (200 mg,
(14). (4aS,5S)-1-(4-Fluorophenyl)-4a-
HeLa or A549 cells were transiently transfected with a lucifer-
ase reporter construct carrying the MMTV promoter which con-
tains two glucocorticoid response elements (GREs). Assays were

C. J. Bungard et al. / Bioorg. Med. Chem. 19 (2011) 7374–7386
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compounds for 16 h and reporter gene activity was measured
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expressed as for the transrepression assay. Data is reported as an
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5.2.4. In Vivo inflammation assay
All animal related procedures were approved by the Institu-
tional Animal Care and Use Committee at Merck Research Labora-
tories, West Point, PA and conform to the Guide for the Care and
Use of Laboratory Animals (US National Institutes of Health,
National Research Council, 1996). Intact adult (6 month old) fe-
male Sprague–Dawley rats are used in the oxazolone (OX) contact
dermatitis model. Rats were sensitized on the ventral abdomen
with OX on Day 0. On Days 7 and 9, a randomly-selected ear was
challenged (same ear each time) with OX; the other was treated
with vehicle. Daily treatment begun on Day 7 and continued for
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Acknowledgments
The authors thank Chuck Ross, Joan Murphy and Jenny Wai for
HRMS analysis and Sandor Varga and Janine Brouillette for NMR
analysis of 11, M1, and M3.
Supplementary data
Supplementary data (details describing the preparation and
characterization of compounds 11, M1, M2, and M3 along with
copies of 1H NMR spectra for all new compounds) associated with
this article can be found, in the online version, at doi:10.1016/
j.bmc.2011.10.054.
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