Heteroaryl and cycloalkyl sulfonamide hydroxamic acid inhibitors of matrix metalloproteinases

Article abstract of DOI:10.1016/S0960-894X(00)00644-2

Heteroaryl and cycloalkyl sulfonamide-hydroxamic acid MMP inhibitors were investigated. Of these, the pyridyl analogue 2 is the most potent and selective inhibitor of MMP-9 and MMP-13 in vitro.

Full text of DOI:10.1016/S0960-894X(00)00644-2

Bioorganic & Medicinal Chemistry Letters 11 (2001) 239±242
Heteroaryl and Cycloalkyl Sulfonamide Hydroxamic Acid
Inhibitors of Matrix Metalloproteinases
Jeremy I. Levin,a,* Yansong Gu,a Frances C. Nelson,a Arie Zask,a John F. DiJoseph,b
Michele A. Sharr,b Amy Sung,b Guixian Jin,a Rebecca Cowling,a Pranab Chanda,b
Scott Cosmi,c Chu-Lai Hsiao,c Wade Edris,c James Wilhelm,c Loran M. Killarb
and Jerauld S. Skotnickia
aWyeth-Ayerst Research, 401 N. Middletown Road, Pearl River, NY 10965, USA
bWyeth-Ayerst Research, PO Box CN-8000, Princeton, NJ 08543, USA
cWyeth-Ayerst Research, Cambridge, MA 02140, USA
Received 7 September 2000; accepted 8 November 2000
AbstractÐHeteroaryl and cycloalkyl sulfonamide-hydroxamic acid MMP inhibitors were investigated. Of these, the pyridyl ana-
logue 2 is the most potent and selective inhibitor of MMP-9 and MMP-13 in vitro. # 2001 Elsevier Science Ltd. All rights reserved.
The normal remodeling of extracellular matrix proteins is
controlled by a family of zinc-containing matrix metallo-
proteinases (MMPs). A variety of pathologies such as
rheumatoid arthritis,1 osteoarthritis,1 and cancer2 have
now been associated with the abnormal regulation of
more than 20 di€erent MMPs. The ability of small
molecule inhibitors of various MMPs to e€ectively treat
arthritis and cancer is now being studied in man. Four
representative compounds to have reached clinical trials
are shown in Figure 1. Marimastat and CG-S27023A
are broad spectrum MMP inhibitors while Ro-32-3555
is selective for the collagenases and AG-3340 shows a
preference for MMP-13 over MMP-1.3
We have recently reported a novel series of anthranilic
acid-based MMP inhibitors of general structure 1, which
di€ers from most other inhibitors in having a two carbon
linker between the sulfonamide nitrogen and the zinc
chelating hydroxamic acid (Fig. 2).4
Figure 2. Anthranilic acid-based MMP inhibitors.
The SAR of the anthranilic acid MMP inhibitors had
shown that the sulfonamide, hydroxamic acid, and a third
substituent must be on adjacent carbons of the aromatic
ring, as shown in Figure 2, to provide compounds with
potent activity against MMP-1, MMP-9, and MMP-13.4
In an e€ort to assess the e€ect of heteroatom substitution,
ring size, and saturation on the anthranilic acid portion
of the molecule we targeted the pyridyl, thienyl, pyr-
azolyl and cycloalkyl analogues of the anthranilic acids.
We now disclose several extensions of this series,
demonstrating that the anthranilic acid phenyl ring can
be replaced with heteroaryl groups as well as a simple
cyclohexyl moiety.
Figure 1. MMP inhibitors in clinical trials.
*Corresponding author. Tel.: +1-845-732-3053; fax: +1-845-732-5561;
e-mail: levinji@war.wyeth.com
0960-894X/01/$ - see front matter # 2001 Elsevier Science Ltd. All rights reserved.
PII: S0960-894X(00)00644-2

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J. I. Levin et al. / Bioorg. Med. Chem. Lett. 11 (2001) 239±242
Chemistry
The pyridyl analogue 2, with the required substitution
pattern, was synthesized in eight steps starting from
3-amino-2,6-dimethoxypyridine, as shown in Scheme 1.5
Thus, reaction of the aminopyridine 3 with BOC-anhy-
dride provided the carbamate 4 in quantitative yield.
Metalation of the pyridine ring ortho to the carbamate
with n-butyllithium and TMEDA followed by the addi-
tion of methyl chloroformate then gave the methyl ester
5 in 34% yield.6 Removal of the BOC moiety was
accomplished with p-toluenesulfonic acid and the result-
ing amine was converted into the p-methoxybenzene
sulfonamide in 86% overall yield. The sulfonamide was
subsequently alkylated with benzyl bromide and sodium
hydride in quantitative yield to give 6. Hydrolysis of the
methyl ester with lithium hydroxide followed by conver-
sion into the corresponding acid chloride and reaction
with hydroxylamine then gave the desired pyridine 2, an
analogue of the anthranilic acid series.
Scheme 2. (i) 4-MeOPhSO2Cl, TEA; (ii) BnBr, NaH; (iii) NaOH; (iv)
(a) (COCl)2, DMF, (b) NH2OH.
(R=Ph) and 11b (R=3-pyridyl) as before. Palladium(II)
catalyzed coupling of bromothiophene 10a with TMS-
acetylene and subsequent desilylation gave 12, which
was converted into hydroxamic acid 13 as before.
Scheme 1. (i) BOC2O; (ii) (a) n-BuLi, TMEDA, (b) ClCO2Me; (iii)
p-TsOH; (iv) 4-MeOPhSO2Cl, TEA; (v) BnBr, NaH; (vi) LiOH; (vii)
(a) (COCl)2, DMF; (b) NH2OH.
The thiophene and pyrazole heteroaryl analogues of the
anthranilic acids were readily available from commercial
starting materials as shown in Scheme 2. The appro-
priate thiophene and pyrazole amino-esters 7a±d were
sulfonylated with p-methoxybenzenesulfonyl chloride
and pyridine to give sulfonamides 8a±d. Alkylation of
the sulfonamides proceeded with sodium hydride and
benzyl bromide. Hydrolysis of the esters and conversion
of the resulting carboxylic acids into the hydroxamic
acids proceeded as before to give 9a±d.
Scheme 3. (i) NBS, AcOH; (ii) BnBr, NaH or 3-PyCH2Cl, K2CO3; (iii)
NaOH; (iv) (a) (COCl)2, DMF; (b) NH2OH; (v) (a) TMS-CCH,
(PPh3)2PdCl2, CuI, (b) Bu4NF.
The saturated analogues of the anthranilic acid MMP
inhibitors were prepared as shown in Scheme 4. Thus,
racemic trans- and cis-2-aminocyclohexane carboxylic
acids, 14a and 14b respectively, were reacted with
p-methoxybenzenesulfonyl chloride to give the analogous
sulfonamide-carboxylic acids. The acids were then con-
verted into the corresponding tert-butyl esters with
N,N-dimethylformamide di-tert-butyl acetal in re¯uxing
toluene. The NH-sulfonamides were alkylated with ben-
zyl bromide and sodium hydride to give sulfonamide-
esters 15. The tert-butyl esters were next deprotected
with tri¯uoroacetic acid to provide the carboxylic acids.
Hydroxamic acid formation was accomplished as before
with oxalyl chloride/DMF followed by the addition of
hydroxylamine to give 16a (trans) and b (cis).
The bromo and alkynyl-substituted thiophenes were syn-
thesized from sulfonamide 8a as shown in Scheme 3. The
alkynes were targeted since they provided a readily acces-
sible substituent roughly equivalent in size to the methyl
group that had enhanced the activity of compound 9c.
Bromination of 8a with N-bromosuccinimide in acetic
acid provided the corresponding bromothiophene in
81% yield. Benzylation as before, or alkylation with
potassium carbonate and 3-picolyl chloride, gave the
esters 10a (R=Ph) and 10b (R=3-pyridyl). Esters 10a
and 10b were then converted into hydroxamates 11a

J. I. Levin et al. / Bioorg. Med. Chem. Lett. 11 (2001) 239±242
241
approximately a 100-fold increase in potency against
MMP-9. The two bromothiophenes, 11a and 11b, were
both moderately potent versus MMP-9. As in the anthra-
nilic acid series, the N-picolyl sulfonamide is somewhat
more active than the N-benzyl derivative. The alkynyl
substituent of thiophene 13 is less e€ective at increasing
potency, perhaps due to its smaller steric bulk. None of
the thiophene hydroxamic acids approached the level of
MMP inhibition achieved by the anthranilic acid hydro-
xamic acids (see 1a, Table 1). The picolyl derivative 11b
was inactive on oral dosing at 100 mg/kg against MMP-9
in the rat dialysis implant bioactivity model.
Scheme 4. (i) 4-MeOPhSO2Cl, TEA; (ii) (Me)2NCH(OtBu)2; (iii)
BnBr, NaH; (iv) TFA; (v) (a) (COCl)2, DMF, (b) NH2OH.
Biology
The pyrazole analogue 9d was only weakly active
against MMP-9 even with the methyl group on the pyr-
azole nitrogen acting as the requisite second substituent
adjacent to the sulfonamide nitrogen. The electron de®-
cient nature of the pyrazole ring may be responsible for
the diminished in vitro activity of 9d relative to the
electron rich thiophenes.
All of the hydroxamic acid ®nal products were tested in
vitro7,8 for their ability to inhibit MMP-9. Inhibitors of
MMP-9 are postulated to have utility as inhibitors of
tumor metastasis. Most compounds were also tested
against the collagenases MMP-1 and MMP-13, which
are presumed to be important in the etiology of osteo-
arthritis. The in vitro potencies of these compounds are
shown in Table 1.
The cyclohexyl analogues 16a and 16b are also less active
than the anthranilic acid hydroxamic acids 1a and 1b ver-
sus both MMP-9 and MMP-13. The trans-cyclohexyl
derivative is more potent than the cis-isomer against
MMP-1, MMP-9, and MMP-13. The level of activity
for the trans-cyclohexyl analogue, 16a, is somewhat
high, however, in light of the fact that the correspond-
ing anthranilic acid analogue lacking a 3-substituent (1,
R1=Bn, R2=H) is a 555 nM inhibitor of MMP-13.
Pyridine analogue 2 is a potent inhibitor of both MMP-9
and MMP-13 with moderate selectivity over MMP-1.
Compounds with this selectivity pro®le may be useful in
assessing the basis of musculoskeletal side e€ects seen in
clinical trials of some MMP inhibitors.9 Pyridine 2 is
almost 3-fold more potent against MMP-13 than the
corresponding 3-methoxy anthranilic acid analogue (1b,
Table 1), and much more selective over MMP-1. The rea-
son for this apparent increase in potency is not known.
Compound 2 is also a moderately potent inhibitor of
TNF-a converting enzyme (TACE), with an IC50 of 294
nM.10 Unfortunately, compound 2 was not orally active
against MMP-13 at a dose of 50mg/kg in the rat dialysis
implant bioactivity model.11
In conclusion, we have investigated the pyridyl, thio-
phene, pyrazole and cyclohexyl analogues of the anthra-
nilate hydroxamate class of MMP inhibitors. The pyridyl
derivative 2 is more potent and selective in vitro than
the related anthranilate 1b. The thiophene and pyrazole
rings proved to be less satisfactory replacements for the
phenyl ring of the anthranilate hydroxamic acids. It is
unclear whether the decrease in potency observed for
these two sca€olds is due to electronic or steric e€ects.
The cyclohexyl ring is also a less e€ective sca€old for the
sulfonamide-hydroxamate pharmacophore, although
incorporating additional substituents on the cycloalkyl
ring might provide more active compounds. Two of the
compounds (2 and 11b) were tested in vivo and were
inactive at the dose tested.
The thiophene derivatives 9a and 9b, lacking a second
substituent adjacent to the sulfonamide nitrogen were
very weakly active versus MMP-9 in accord with the SAR
previously shown for the anthranilic acid derivatives. As
expected, incorporation of a methyl substituent next to
the sulfonamide nitrogen in thiophene 9c provides
Table 1. In vitro potency of sulfonamide-hydroxamic acids
Compound
MMP-1a
MMP-9a
MMP-13a
References and Notes
1a (R1=CH2-3-Py, R2=Me)
1b (R1=CH2Ph, R2=OMe)
2
9a
9b
9c
9d
11a
11b
13
16a
16b
143
520
1227
5
23
15
8
138
47
NTc
NT
NT
NT
427
290
1. (a) Clark, I. M.; Rowan, A. D.; Cawston, T. E. Curr. Opin.
Anti-In¯ammat. Immunomodulat. Investigat. Drugs 2000, 2, 16.
(b) Shaw, T.; Nixon, J. S.; Bottomley, K. M. Exp. Opin.
Invest. Drugs 2000, 9, 1469. (c) Bottomley, K. M.; Johnson,
W. H.; Walter, D. S. J. Enzyme Inhib. 1998, 13, 79.
2. (a) Yip, D.; Ahmad, A.; Karapetis, C. S.; Hawkins, C. A.;
Harper, P. G. Investigat. New Drugs 1999, 17, 387. (b) Nelson,
A. R.; Fingleton, B.; Rothenberg, M. L.; Matrisian, L. M. J.
Clin. Oncol. 2000, 18, 1135.
3. (a) For recent comprehensive reviews, see: Montana, J.;
Baxter, A. Curr. Opin. Drug Disc. Devel. 2000, 3, 353. (b)
Whittaker, M.; Floyd, C. D.; Brown, P.; Gearing, A. J. H.
Chem. Rev. 1999, 99, 2735. (c) Beckett, R. P.; Whittaker, M.
40% (1)b
19% (1)b
NT
67% (10)b
57% (10)b
104
22% (1)b
639
900
236
70
50% (1)b
49% (1)b 38% (0.3)b 35% (0.3)b
174
361
181
318
233
291
aIC50 (nM).
b% Inhibition (mM).
cNT=not tested.

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J. I. Levin et al. / Bioorg. Med. Chem. Lett. 11 (2001) 239±242
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7. Weingarten, H.; Feder, J. Anal. Biochem. 1985, 147, 437.
8. Inhibitor concentrations were run in triplicate and IC50
determinations were calculated from a four-parameter logistic
®t of the data within a single experiment.
9. Scrip. 1998, 2349, 20.
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A.; Cowling, R. Manuscript in preparation.
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