MMP-13 selective isonipecotamide α-sulfone hydroxamates

Article abstract of DOI:10.1016/j.bmcl.2010.04.111

A series of N-aryl isonipecotamide α-sulfone hydroxamate derivatives has been prepared utilizing a combination of solution-phase and resin-bound library technologies to afford compounds that are potent and highly selective for MMP-13.

Full text of DOI:10.1016/j.bmcl.2010.04.111

Bioorganic & Medicinal Chemistry Letters 20 (2010) 3561–3564
Contents lists available at ScienceDirect
Bioorganic & Medicinal Chemistry Letters
journal homepage: www.elsevier.com/locate/bmcl
MMP-13 selective isonipecotamide
a-sulfone hydroxamates
Stephen A. Kolodziej, Susan L. Hockerman, Gary A. DeCrescenzo, Joseph J. McDonald, Debbie A. Mischke,
,
*
Grace E. Munie, Theresa R. Fletcher, Nathan Stehle, Craig Swearingen, Daniel P. Becker
Departments of Medicinal Chemistry and Pharmacology, Pfizer Research & Development, 700 Chesterfield Village Parkway, St. Louis, MO 63198, 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 N-aryl isonipecotamide a-sulfone hydroxamate derivatives has been prepared utilizing a com-
bination of solution-phase and resin-bound library technologies to afford compounds that are potent and
highly selective for MMP-13.
Received 11 March 2010
Revised 23 April 2010
Accepted 26 April 2010
Available online 28 April 2010
Ó 2010 Elsevier Ltd. All rights reserved.
Keywords:
Matrix metalloproteinase inhibitor
MMP inhibitor
MMP-13 selective
Hydroxamate
Metalloenzyme
Matrix metalloproteinases (MMPs) are zinc-dependent en-
zymes that are responsible for remodeling and degradation of all
components of the extracellular matrix,1,2 yet excessive activity
of MMPs has been implicated in numerous disease states including
cancer,3,4 arthritis,5 and cardiovascular disease.6–9 MMP inhibitors
(MMPi’s) have therefore been explored as therapeutic treatments
to halt progression of various diseases.10–12 The MMP family of en-
zymes includes at least 24 distinct mammalian isozymes, but
MMP-13 in particular has been identified as a significant target
since its upregulation has been implicated in cancer, osteoarthritis,
and cardiovascular disease.
Treatment of patients with broad-spectrum MMPi’s gives rise to
stiffening of the joints referred to as musculoskeletal syndrome9
(MSS). Inhibition of MMP-1 has been hypothesized to be the cause
of MSS observed clinically with broad-spectrum inhibitors, and the
broad-spectrum inhibitor marimastat induces musculoskeletal
side effects in rats.13 MMP-1 has long been suspected as a culprit
whose inhibition plays a role in MSS. In addition, MT-1 MMP
(MMP-14) knockout mice suffer connective tissue disease due to
inadequate collagen turnover14 and impaired endochondral ossifi-
cation15 reminiscent of joint lesions in MSS. We have therefore
concentrated our efforts on potently inhibiting MMP-13 while
sparing other MMPs to achieve joint safety, in particular MMP-1
and MMP-14, which we refer to as the dual-sparing hypothesis.
researchers.16–18 Moderately selective pyrimidinetrione MMP-13
inhibitors have been reported that gave rise to fibroplasia in a
14-day rat study, but MMP-14 data was not reported.19
We previously described the synthesis and MMP inhibitory
activity of b-sulfone hydroxamates20,21 and aryl-linked isoster-
es22,23 that potently inhibit MMP-2 and MMP-13 but spare MMP-
1, and discovered that
a-sulfone hydroxamates including SC-276
are superior to the b-sulfones in both MMP-1 sparing enzyme pro-
files and ADME properties, and exhibit excellent oral antitumor
efficacy in vivo.24 MMP-1 sparing
a-sulfone hydroxamates have
also been reported by the Wyeth group through modification of
P10 substituents,25,26 and Wyeth researchers have also employed
b-sulfones to attain potent and selective TACE inhibitors.27,28
Zhang et al. of J&J have employed
a-sulfone carboxylic acids as
MMP-1 sparing gelatinase (MMP-2/9) inhibitors.29 Our work in
exploring modifications in the P0 region toward further enhancing
MMP-13 selectivity through interaction with the S01 pocket has
afforded a series of aryl piperidines and isonipecotamide deriva-
tives that are highly selective for MMP-13 and sparing of both
MMP-1 and MMP-14 as we report herein.
Isonipecotamide sulfone hydroxamates 4 in the
a-tetrahydro-
pyran series were prepared as outlined in Scheme 1. Carboxylic
acid 124 was coupled with the hydroxamate-containing modified
Wang resin of Floyd30 employing benzotriazol-1-yl-oxytripyrroli-
dinophosphonium hexafluorophosphate (PyBOP) as the coupling
agent with N-methylmorpholine (NMM) in N-methylpyrrolidinone
(NMP) to give polymer-bound aryl fluoride 2. Nucleophilic aro-
matic substitution with a 10-fold excess of ethyl isonipecotate in
NMP, and subsequent hydrolysis of the ethyl ester gave resin-
bound carboxylic acid 3. The polymer-bound acid was activated
MMP-13 selective a-carboxylic acids have been reported by Wyeth
* Corresponding author. Tel.: +1 773 508 3089; fax: +1 773 508 3086.
E-mail address: dbecke3@luc.edu (D.P. Becker).
Present address: Department of Chemistry, Loyola University, 6525 North
Sheridan Road, Chicago, IL 60626, USA.
0960-894X/$ - see front matter Ó 2010 Elsevier Ltd. All rights reserved.
doi:10.1016/j.bmcl.2010.04.111

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S. A. Kolodziej et al. / Bioorg. Med. Chem. Lett. 20 (2010) 3561–3564
Scheme 1. Synthesis of isonipecotamide sulfone hydroxamates in the a-tetrahydropyran series.
with PyBOP and reacted with the requisite amine to give the corre-
sponding polymer-bound amides, which were liberated from the
resin with TFA to afford isonipecotamides 4.
a-piperidines were as potent as a-tetrahydropyrans in the broad-
er-spectrum, MMP-1 sparing series,24 while the potency for
MMP-2 increased modestly to 1700 nM resulting in a 50-fold drop
in selectivity versus MMP-2. cis-Dimethylmorpholine 4h was 4X
less potent than 4f, suggesting that the trans isomer may be the
more potent isomer in 4f. Piperazine amides 4i–4n suffered a loss
of potency for MMP-13, particularly with the introduction of a ba-
sic amine leading to the least potent analog 4n. N-Aryl piperazine
amides 4o–4w in general were more potent for MMP-13 with good
selectivities versus MMP-2. Fluoro analogs 4o and 4q were among
the most potent analogs (IC50 = 6.7 and 6.0 nM, respectively), along
with 4-acyl derivative 4r. The 2,4-dimethylphenyl analog 4s main-
tained decent potency for MMP-13 (IC50 = 12.2 nM) and was less
potent at MMP-2 leading to a selectivity of 460X. MMP-13 toler-
ated heterocyclic analogs 4t–4w with a nitrogen in the 2-position
of 4t and 4u (IC50 = 10.7 and 6.4 nM, respectively) with good selec-
tivities (330X and 300X, respectively), whereas a nitrogen in the 3-
or 4-position led to a loss of some potency and selectivity (4v and
4w).
Isonipecotamides 4 in the a-piperidine sulfone series were pre-
pared by traditional solution-phase methodologies as outlined in
Scheme 2. Ethyl isonipecotate N-tert-butylcarbamate 531 was cou-
pled with the requisite amine using 2-chloro-4,6-dimethoxy-1,3,5-
trazine (CDMT) as a coupling reagent followed by deprotection with
HCl to afford piperidine 6. Nucleophilic aromatic displacement of
aryl fluoride 724 gave the aryl piperidine sulfone 8. Hydrolysis of
the ethyl ester, coupling with THP-protected hydroxylamine using
EDC and HOBT followed by acidic deprotection afforded the
hydroxamates 4 as the
a
-piperidine hydrochloride salts.
-tetrahydropyranyl and
The inhibitory potencies of
a
a-piperi-
dine sulfone hydroxamates 4a–w versus MMP-2 and MMP-13 are
summarized in Table 1 wherein the isonipecotic acid amide moiety
was varied. Also shown in Table 1 is a selectivity ratio derived from
dividing the IC50 at MMP-2 by that of MMP-13. Moderate potencies
for MMP-13 were maintained, and single-digit nanomolar potency
was attained for several analogs. All compounds had IC50 values of
>10,000 nM for MMP-1 (not shown), thus selectivities for MMP-13
versus MMP-1 varied from >100X (4n) to >2000X. Selectivity ratios
versus MMP-2 were generally in a range of 50–500, and as high as
1659 for 4f. Allyl and propargyl derivatives 4a and 4b were moder-
ately potent for MMP-13 with selectivities versus MMP-2 of
approximately 85X. Selectivities rose for aralkyl substituted deriv-
atives 4c, 4d, and 4e to nearly 400X for 4e. 3,5-Dimethylpiperidine
amide 4f (mixture of cis and trans isomers) distinguished itself as
the most potent for MMP-13 (IC50 = 4.4 nM) and the most selective
Table
2
summarizes inhibitory potency for 2,3-dim-
-tetrahydropyr-
ethylphenylpiperidine amides 4x, 4y, and 4z. The
a
anyl (X = O) compound 4x distinguished itself as both the most
potent and selective of the isonipecotic amides, with an IC50 for
MMP-13 of 4.0 nM and selectivity of 40X versus MMP-3, 1500X
versus MMP-2, and >2500X versus MMPs-1, 8, 9, and 14. Unfortu-
nately, this compound was below the detection level when dosed
orally in rats. The corresponding N-cyclopropyl and N-methoxy-
ethyl piperidine analogs 4y and 4z were thus prepared, but the
MMP-13 inhibitory potency for these compounds dropped 7X
and 17X, respectively.
versus MMP-2 as well (1659X). The corresponding a-piperidine N-
methoxyethyl 4g analog was prepared to improve aqueous solubil-
ity and ADME properties relative to 4f (X = O). Surprisingly the
MMP-13 potency for 4g dropped to an IC50 of 50 nM, although
Table 3 shows the MMP inhibitory and rat PK data for aniline
amide 4aa, which had good potency for MMP-13 (IC50 = 9.0 nM)
and very good selectivities versus both MMP-1 and MMP-14
Scheme 2. Synthesis of isonipecotamide sulfone hydroxamates in the a-piperidine series.

S. A. Kolodziej et al. / Bioorg. Med. Chem. Lett. 20 (2010) 3561–3564
3563
Table 1
MMP inhibitory potency of isonipecotamide
a
-tetrahydropyranyl and
a
-piperidine sulfone hydroxamates 4a–wa
Compd
X
NR1R2
IC50 (nM) at MMP-Xa
Selectivity ratio 2/13
2
13
35
4a
4b
4c
4d
4e
4f
4g
4h
4i
4j
4k
4l
4m
4n
4o
4p
4q
4r
O
O
Allyl(methyl)amino
Methyl(prop-2-ynyl)amino
Benzyl(methyl)amino
3,4-Dihydroisoquinolin-2(1H)-yl
6,7-Dimethoxy-3,4-dihydroisoquinolin-2(1H)-yl
3,5-Dimethylpiperidin-1-yl
3,5-dimethylpiperidin-1-yl
cis-2,6-Dimethylmorpholin-4-yl
4-Acetylpiperazin-1-yl
2900
83
86
2100
24.5
20
9
6.2
4.4
50
18.1
50
28
45
25.1
40
90
6.7
18.0
6.0
6.7
12.2
10.7
6.4
30
N-Cyclopropyl
O
O
O
3500
2100
2400
7300
1700
>10K
2500
5500
7000
900
6000
>10,000
1400
4500
1600
700
175
233
387
1659
34
552
50
196
156
36
150
>111
209
250
267
104
459
336
250
58
N-CH2CH2OMe
O
O
O
O
O
O
O
O
O
O
O
O
O
O
O
O
4-Isopropylpiperazin-1-yl
4-(2-Methoxyethyl)piperazin-1-yl
4-Phenethylpiperazin-1-yl
4-(2-Hydroxyethyl)piperazin-1-yl
4-(2-(Dimethylamino)ethyl)piperazin-1-yl
4-(2-Fluorophenyl)piperazin-1-yl
4-(2-Methoxyphenyl)piperazin-1-yl
4-(4-Fluorophenyl)piperazin-1-yl
4-(4-Acetylphenyl)piperazin-1-yl
4-(2,4-Dimethylphenyl)piperazin-1-yl
4-(Pyridin-2-yl)piperazin-1-yl
4-(Pyrimidin-2-yl)piperazin-1-yl)
4-(Pyridin-4-yl)piperazin-1-yl
4-(Pyrazin-2-yl)piperazin-1-yl
4s
4t
4u
4v
4w
5600
3600
1600
1750
900
26.8
34
a
MMP-1 IC50 >10,000 nM for all compounds.
Table 2
MMP inhibitory potency of isonipecotamide
a
-sulfone hydroxamates 4x, 4y, and 4z
Compd
X
IC50 (nM) at MMP-X
8
1
2
3
9
13
4.0
27.7
70
14
4x
4y
4z
O
>10,000
>10,000
>10,000
6000
6350
400
160
550
—
>10,000
9400
—
>10,000
>10,000
—
>10,000
>10,000
—
N-Cyclopropyl
NCH2CH2OMe
Table 3
MMP inhibitory potency and rat PK data of isonipecotamide
a
-sulfone hydroxamate 4aa
Compd
clog P
IC50 (nM) at MMP-X
Rat PK
1
2
3
8
9
13
9.0
14
Cmax
C6h
t1/2 (h)
0.83
BA (%)
4.2
4aa
1.0
>10,000
400
370
>10,000
1230
>10,000
1490
20

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(>1100X). However, exposure and half life in the rat were very poor
after oral dosing, with a half life of less than 1 h, and a BA of only
4%. Isonipecotamide hydroxamates described herein have demon-
strated double-digit to single-digit potency for MMP-13 combined
with very good selectivity versus MMP-1 (110–2500) and versus
MMP-2 ranging from 30X to 1500X. Compound 4x exhibits
>2500X selectivity for MMP-13 versus both MMP-1 and MMP-14,
hence we refer to this profile as dual-sparing (e.g., MMP-13 po-
tency while sparing both MMP-1 and MMP-14). Yet rat PK for
4aa was disappointing, but not surprising with a high molecular
weight of 544 a.u.32 We therefore turned our attention to lower
molecular weight species, while applying our learnings about P10
manipulations toward optimizing MMP-13 selectivity and ulti-
mately to MMP-1/14 dual-sparing profiles with lower MW and
fewer reduce rotatable bonds as described in the subsequent
publication.33
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