Development of new hydroxamate matrix metalloproteinase inhibitors derived from functionalized 4-aminoprolines

Article abstract of DOI:10.1021/jm000246e

A series of hydroxamates was prepared from an aminoproline scaffold and tested for efficacy as matrix metalloproteinase (MMP) inhibitors. Detailed SAR for the series is reported for five enzymes within the MMP family, and a number of inhibitors, such as compound 47, display broad-spectrum activity with sub-nanomolar potency for some enzymes. Modifications of the P1′ portion of the molecule played a key role in affecting both potency and selectivity within the MMP family. Longer-chain aliphatic substituents in this region of the molecule tended to increase potency for MMP-3 and decrease potency for MMP-1, as exemplified by compounds 48-50, while aromatic substituents, as in compound 52, generated broad-spectrum inhibition. The data is rationalized based upon X-ray crystal data which is also presented. While the in vitro peroral absorption seemed to be less predictable, it tended to decrease with longer and more hydrophilic substituents. Finally, a rat model of osteoarthritis was used to evaluate the efficacy of these compounds, and a direct link was established between their pharmacokinetics and their in vivo efficacy.

Full text of DOI:10.1021/jm000246e

4948
J . Med. Chem. 2000, 43, 4948-4963
Develop m en t of New Hyd r oxa m a te Ma tr ix Meta llop r otein a se In h ibitor s Der ived
fr om F u n ction a lized 4-Am in op r olin es
Michael G. Natchus,* Roger G. Bookland, Biswanath De, Neil G. Almstead, Stanislaw Pikul, Michael J . J anusz,
Sandra A. Heitmeyer, Erin B. Hookfin, Lily C. Hsieh, Martin E. Dowty, Charles R. Dietsch, Vikram S. Patel,
Susan M. Garver, Fei Gu, Matthew E. Pokross, Glen E. Mieling, Timothy R. Baker, David J . Foltz,
Sean X. Peng, David M. Bornes, Michael J . Strojnowski, and Yetunde O. Taiwo
Procter and Gamble Pharmaceuticals, 8700 Mason-Montgomery Road, Mason, Ohio 45040
Received J une 8, 2000
A series of hydroxamates was prepared from an aminoproline scaffold and tested for efficacy
as matrix metalloproteinase (MMP) inhibitors. Detailed SAR for the series is reported for five
enzymes within the MMP family, and a number of inhibitors, such as compound 47, display
broad-spectrum activity with sub-nanomolar potency for some enzymes. Modifications of the
P1′ portion of the molecule played a key role in affecting both potency and selectivity within
the MMP family. Longer-chain aliphatic substituents in this region of the molecule tended to
increase potency for MMP-3 and decrease potency for MMP-1, as exemplified by compounds
48-50, while aromatic substituents, as in compound 52, generated broad-spectrum inhibition.
The data is rationalized based upon X-ray crystal data which is also presented. While the in
vitro peroral absorption seemed to be less predictable, it tended to decrease with longer and
more hydrophilic substituents. Finally, a rat model of osteoarthritis was used to evaluate the
efficacy of these compounds, and a direct link was established between their pharmacokinetics
and their in vivo efficacy.
In tr od u ction
The matrix metalloproteinases (MMPs) are a family
of enzymes that are intimately involved in tissue
remodeling. They have thus received a great deal of
detailed attention,1 and inhibitors for these enzymes
have been developed for the treatment of a startlingly
wide array of disease processes where matrix remodel-
ing plays a key role. These indications include osteo-
arthritis,2-4 rheumatoid arthritis,3,5,6 tumor metastasis,7
multiple sclerosis,8-11 congestive heart failure,11-13 and
a host of others.
A number of MMPIs have progressed into clinical
trials for cancer, rheumatoid arthritis, and possibly
osteoarthritis; selected examples are shown in Figure
1.14 Marimastat is a broad-spectrum inhibitor and was
the first MMPI to enter clinical trials for cancer treat-
ment.15 So far, progression through these trials has been
hampered due to musculoskeletal syndrome (MSS)
F igu r e 1. Clinical candidates that inhibit MMPs.
which manifests itself as musculoskeletal pain after a
series of inhibitors based on an aminopyrrolidine scaf-
few weeks of treatment. One hypothesis suggests that
fold was developed which displayed a wide range of
inhibition of MMP-1 may play a role in the appearance
enzyme profiles and is described below. Further inves-
of MSS. Consequently, recent efforts in the field have
tigation led to the identification of a number of selective,
been directed toward designing MMP-1-sparing inhibi-
orally active inhibitors which protected cartilage against
tors. Out of this new generation, BA-129566 emerged
degradation in a rat model of osteoarthritis.
as a highly selective inhibitor which reportedly showed
no signs of MSS in phase 2 clinical trials.2
Syn th esis
Our group has been developing both succinamide16
All compounds were derived from cis-4-hydroxy-D-
proline (1) as exemplified in Scheme 1. The presence of
a hydroxyl group at C4 provided a functionalizable
and heterocyclic17-20 inhibitors with a focus on under-
standing the structural modifications which might
deliver the appropriate inhibitory profile which will slow
handle from which various moieties were obtained
the progression of osteoarthritis while minimizing or
stereospecifically. Mesylates of type 2 were obtained via
a conventional sequence of esterification, sulfonylation,
eliminating the occurrence of MSS. From this effort, a
and treatment with methanesulfonyl chloride. This key
* To whom correspondence should be addressed. Tel: 513-622-3646.
Fax: 513-622-3681. E-mail: natchus.mg@pg.com.
intermediate was isolated and stored for long periods
10.1021/jm000246e CCC: $19.00 © 2000 American Chemical Society
Published on Web 11/10/2000

New MMP Inhibitors from Functionalized Aminoprolines
J ournal of Medicinal Chemistry, 2000, Vol. 43, No. 26 4949
Sch em e 1a
a
Reagents: (a) MeOH, SOCl2; (b) ClSO2C6H4OMe; (c) MsCl, NEt3, CH2Cl2; (d) NaN3, DMF, 60 °C; (e) 10% Pd-C, H2, EtOAc; (f) NH2OH,
MeOH; (g) CH3CH2CHO, NaCNBH3, NaOAc; (h) ClSO2Me, NEt3, CH2Cl2; (i) HCO2H, (CH3OCO)2O; (j) BH3, THF.
Sch em e 2a
a
Reagents: (a) 3-N-methylhydantoin, PPh3, DEAD, CH2Cl2; (b) NH2OH, KOH, MeOH; (c) MsCl, NEt3, CH2Cl2; (d) NaN3, DMF, 60 °C;
(e) 10% Pd-C, H2, EtOAc; (f) (BrCH2CH2)2O, Et3N, DMF, 65 °C; (g) Cl(CH2)3SO2Cl, NEt3, CH2Cl2; (h) DBU, DMF.
of time and was resistant to elimination even under
mildly basic conditions. The mesylate can then be
readily displaced upon treatment with sodium azide.
Subsequent hydrogenation over 10% Pd-C gave the
versatile primary amine 3 in high yield as a single
diastereomer.
The free primary amine allowed for a diverse array
of functionalization. Reductive amination with alkyl-
aldehydes such as propanal gave monoalkylation prod-
ucts of type 4. Mixtures of monoalkylation and both
bisalkylation products were obtained with formalde-
hyde. Monomethylation was achieved by first forming
a formamide with free amine 3 and then reduction with
BH3‚THF. Acyl groups were coupled with either the
primary amine 3 or the alkylamine 4 to give a wide
range of functionality such as amides, sulfonamides, and
ureas. Sulfonamides 5-7 are typical examples. The
methyl esters in these derivatives were then converted
to their respective hydroxamic acids (e.g. 19, 21, 23, 26,
and 27) upon treatment with hydroxylamine.
Heterocyclic structures were introduced at C4 of the
pyrrolidine ring by direct substitution or via function-
alization of primary amine 3 as seen in Scheme 2.
Treatment of alcohol 8 with functionalized hydantoins
under Mitsunobu conditions provided clean substitution
at C4 to give heterocyclic compounds of type 9. Direct
displacement of mesylates such as 2 with secondary
amines such as morpholine was also possible, but
tertiary amines of type 10 could be explored much more
efficiently via primary amines of type 3. Cycloalkylation
was carried out cleanly upon heating 3 with dibromoal-
kanes such as dibromoethyl ether in DMF which af-
forded compound 10. Additionally, cyclic amides and
sulfonamides were prepared upon acylation of 3 with
haloalkylacyl chlorides and haloalkylsulfonyl chlorides
of type 11. Subsequent exposure to DBU in DMF then

4950 J ournal of Medicinal Chemistry, 2000, Vol. 43, No. 26
Natchus et al.
Sch em e 3a
a
Reagents: (a) MsCl, NEt3, CH2Cl2; (b) NaN3, DMF, 60 °C; (c) 10% Pd-C, H2, EtOAc; (d) (BrCH2CH2)2, Et3N, DMF, 65 °C; (e) TFA,
CH2Cl2; (f) PhOC6H4SO2Cl, NEt3, CH2Cl2; (g) Cl(CH2)3SO2Cl, NEt3, CH2Cl2; (h) t-BuOK, THF; (i) NH2OH, MeOH.
afforded clean cyclization to give structures of type 12.
The methyl esters of these hyterocyclic structures were
then converted to their respective hydroxamic acids
upon treatment with hydroxylamine.
selective compounds which strongly inhibit MMP-2 and
-13 while sparing MMP-7. On the other hand, the
inhibition of MMP-1 and -3 varied much more broadly.
While MMP-3 was generally inhibited more potently
than MMP-1, selectivities between these enzymes were
much more sensitive to functional group manipulation.
Su bstitu ted Am in e SAR. The C4 amine functional-
ity tolerated diverse monosubstitution with remarkably
little effect on potency as seen in Table 1 with the
comparison of free amine 19 against n-propylamine 21,
sulfonamides 23 and 29, and lactic acid amide 37. Only
the urea substitution in 32 seemed to be poorly tolerated
and resulted in 1 order of magnitude loss in potency for
all enzymes tested. The potencies of the N-sulfonamides
increased for MMP-1 and -3 upon N-alkylation as seen
with 23 vs 26-28. A similar potency boost was not
observed with amide 33 or any of the lactic acid amides
in Table 2.
All of the substituted amines in Tables 1 and 2 appear
to be sensitive to modifications of the S1′ portion of the
molecule (R). This sensitivity is typified by free amines
19 and 20, sulfonamides 23 and 24, and amides 34 and
35 where longer-chain substituents increase the potency
for MMP-3 and decrease the potency for MMP-1. This
observation is supported by X-ray data which place the
alkoxyaryl group in the S1′ pocket of the MMP enzymes.
This pocket is very deep for most enzymes, but it is
blocked by an arginine and a tyrosine residue in MMP-1
and -7, respectively.23 As a result, the selectivity for
MMP-3 vs MMP-1 increased from 10-fold, which is
typical for a methoxy substituent, to 50-100-fold, which
is typical for an n-butoxy substituent.
An alternate synthetic method shown in Scheme 3
was developed to allow for rapid SAR development of
the sulfonamide group. Protection of the ring amine as
a tert-butylcarbamate allowed the C4 substituent to be
functionalized as desired followed by subsequent gen-
eration of a wide range of different sulfonamides. The
protected alcohol 13 was available in two steps from cis-
hydroxy-D-proline via esterification and treatment with
di-tert-butoxycarbonyl anhydride. The primary amine
14 could then be prepared via a sequence of mesylation,
displacement with sodium azide, and hydrogenation
over 10% Pd-C. Cycloalkylation of 14 could be ac-
complished with a variety of dibromoalkanes to give
tertiary amines of type 15. Similarly, cyclic sulfonam-
ides of type 17 were prepared on protected scaffolds as
described for sulfonamide 12. Variously substituted
sulfonamides were then prepared upon deprotection of
the secondary amine followed by sulfonation with a
variety of sulfonyl chlorides including the n-propoxy-
benzene derivative which was used for the preparation
of compounds 18 and the phenoxybenzene derivative
which was used for the preparation of compound 16.
Treatment with hydroxylamine then completed the final
transformation of the methyl esters to the corresponding
hydroxamic acids.
Resu lts a n d Discu ssion
SAR An a lysis. All compounds were assayed in vitro
for the inhibition of truncated collagenase-1 (MMP-1),21
gelatinase-A (MMP-2),21 stromelysin (MMP-3),22 matril-
ysin (MMP-7),21 and collagenase-3 (MMP-13).21 Several
general SAR observations can be made which apply
broadly to most of the aminoprolines described in this
paper. The class in general inhibits MMP-2 and -13 very
potently with many examples showing sub-nanomolar
potency and only a select few showing >10 nM potency.
MMP-7 on the other hand was poorly inhibited relative
to the other enzymes with only a few examples of sub-
micromolar potency. The aminoproline structure is
therefore an excellent scaffold from which to design
C4 Heter ocycle SAR. The heterocyclic compounds
in Tables 3 and 4 displayed inhibition profiles which
closely mimicked those of the substituted amines. The
heterocycles which were bonded through a basic tertiary
amine, such as 44, 48, and 53, provided consistent
profiles regardless of the ring size or extra heteroatoms
within the ring. Also, like the substituted amines, these
compounds were very sensitive to modifications of the
P1′ substituent (R). Compounds 48-50 exemplify the
increase in selectivity for MMP-3 over MMP-1 that is
commonly observed as longer alkoxy chains are intro-
duced. Sultams of type 58 showed a significant potency

New MMP Inhibitors from Functionalized Aminoprolines
J ournal of Medicinal Chemistry, 2000, Vol. 43, No. 26 4951
Ta ble 1. Substituted Amine-Bearing MMP Inhibitors
IC50 (nM)a
no.
Q
W
R
MMP-1
MMP-2
MMP-3
MMP-7
MMP-13
abs %b
19
20
21
22
23
24
25
26
27
28
29
30
-H
-H
-H
-H
-H
-H
-H
-H
-H
-Me
-OMe
-OnBu
-OMe
-OMe
-OMe
-OnBu
-O-4-Pyr
-OMe
-OMe
-OMe
-OMe
-OnBu
250
380
150
560
260
1300
450
19
74
33
350
700
6
nd
nd
1
23
8
4300
nd
5
nd
5
1
2
0.7
0.8
0.5
1
0.8
2
9-39
2
-(CH2)2CH3
-(CH2)2Ph
-SO2CH3
87
57
42
16
22
6
2200
1400
2700
3900
>10000
520
1
1
44
23
nd
nd
40-42
11-13
2
2
-SO2CH3
0.8
nd
nd
2
0.8
3
-SO2CH3
-SO2CH3
-SO2CH3
-CH2-3-Pyr
-CH2CH2CH3
-H
6
2700
920
3000
nd
-SO2CH3
10
36
6
-SO2-p-C6H4OMe
-H
nd
nd
13-22
31
32
33
34
35
-CO-p-Ph-Ph
-CONHMe
-COiPr
-COCH2OMe
-COCH2OMe
-H
-OMe
-OMe
-OMe
-OnPr
-OnBu
>1000
3100
270
500
1200
nd
nd
2.2
nd
nd
400
300
10
30
14
nd
nd
nd
nd
27
17
3
nd
nd
nd
nd
2
-H
-CH2-3-Pyr
-H
-H
>10000
4900
0.7
a
See Experimental Section for details of experimental assays. Standard deviations for enzyme assays were typically (60% of the
b
mean or less. Data obtained from both single and multiple determinations; for experimental details see ref 19. nd, not determined.
Ta ble 2. MMP Inhibitors with Lactic Acid Amide Derivatives
IC50 (nM)a
no.
W
Y
Z
R
MMP-1
MMP-2
MMP-3
MMP-7
MMP-13
abs %b
36
37
38
39
40
41
42
43
-H
-CH3
-CH2Ph
-CH2Ph
-H
-OMe
-OMe
-OMe
-OMe
-OnPr
-OnBu
-OnBu
-OnBu
140
240
87
260
140
1200
270
440
0.3
0.2
0.7
0.6
nd
nd
nd
nd
7
10
2
12
8
10
8
10
nd
nd
8500
nd
3500
4200
1700
2400
nd
2
1
0.5
1
0.9
0.8
0.9
19-23
nd
13-18
7-8
nd
8-11
3
3
-H
-CH2Ph
-CH3
-nPr
-nPr
-H
-CH2CH2Ph
-CH3
-H
-H
-H
-CH3
-H
-H
-iPr
-H
-H
-CH2iPr
-H
a
See Experimental Section for details of experimental assays. Standard deviations for enzyme assays were typically (60% of the
b
mean or less. Data obtained from both single and multiple determinations; for experimental details see ref 19. nd, not determined.
boost consistent with that observed with alkyl sulfon-
amides such as 26. Compounds of this type displayed a
5-fold potency boost for both MMP-1 and -7. However,
good selectivity for MMP-3 vs MMP-1 seemed to be
unattainable as seen with compounds 58-60.
The hydantoins shown in Table 4 presented potency
profiles which were very similar to those seen with the
heterocyclic amines in Table 3. Substitution of the
hydantoin moiety played only a minor role in the overall
potency of the compounds, as seen with 64, 70, and 72.
Substitution in the S1′ region of the molecule (R)
reflected trends observed with other classes of mol-
ecules, and attempts to introduce hydrophilicity into this
portion of the molecule resulted in a significant loss of
potency (see 64 vs 66). This lends support to the
observation that the deep S1′ pocket is a lipophilic
enviornment.23
Str u ctu r e of In h ibitor -Str om elysin Com p lex.24
A crystal structure of stromelysin was obtained with
compound 24 bound in the active site, and the data is
represented graphically in Figures 2 and 3. The struc-
tural data is in agreement with those reported previ-
ously for similar compounds.18,19 The sulfonamide por-
tion of the molecule clearly fits into the deep, hydrophobic
S1′ pocket, and the hydroxamic acid is bound to the
active site zinc atom in fashion. We also see key
enzyme-substrate atomic distances that support the
existence of hydrogen-binding interactions that are
likely to be present in all of the inhibitors in this report.
These include hydrogen bonds from the hydroxamic acid
moiety to Ala 165 and Glu 202 and key hydrogen bonds
from the sulfonamide oxygen to Ala 165 and Leu 164.
A hydrogen bond also appears to exist between His 211
and the methane sulfonamide oxygen which is likely

4952 J ournal of Medicinal Chemistry, 2000, Vol. 43, No. 26
Ta ble 3. MMP Inhibitors Containing Heterocyclic Scaffolds
Natchus et al.
IC50 (nM)a
MMP-3
no.
n
Y
X
R
MMP-1
MMP-2
MMP-7
MMP-13
abs %b
44
45
46
47
48
49
50
51
52
53
54
55
56
57
58
59
60
2
2
2
2
2
2
2
2
2
2
2
1
1
1
1
1
1
CH2
CH2
CH2
CH2
CH2
CH2
CH2
CH2
CH2
CH2
CH2
CH2
CH2
CH2
SO2
SO2
SO2
CH2
CH2
CH2
CH2
O
O
O
O
O
SO2
SO2
CH2
CH2
CH2
CH2
CH2
CH2
-OMe
280
210
2000
17
220
490
920
2000
19
300
>1000
310
20
6
nd
nd
nd
6
nd
4
nd
nd
nd
7
<1
nd
nd
nd
nd
nd
48
9
12
8
14
7
5
12
7
23
12
8
4
5
13
8
13
7400
620
9800
610
1
100
nd
nd
nd
24-39
12-15
3-10
nd
nd
nd
13-17
33-48
3
2-3
16-25
5-6
2-4
-OnPr
-nPent
-OPh
0.4
0.9
<0.4
nd
-OMe
-OnPr
-OnBu
-nPent
-OPh
-OMe
-OnBu
-OnPr
-O-4-C6H4F
-OPh
nd
16500
5400
7400
710
7100
3400
8700
850
1
0.6
0.7
<0.4
1
0.9
1
<0.4
<0.4
0.4
<0.4
<0.4
19
42
100
230
790
1400
840
-OMe
-OnPr
-OnBu
680
a
See Experimental Section for details of experimental assays. Standard deviations for enzyme assays were typically (60% of the
b
mean or less. Data obtained from both single and multiple determinations; for experimental details see ref 19. nd, not determined.
Ta ble 4. MMP Inhibitors Containing Hydantoin Moieties
IC50 (nM)a
no.
Q
X
R
MMP-1
MMP-2
MMP-3
MMP-7
MMP-13
abs %b
61
62
63
64
65
66
67
68
69
70
71
72
73
74
75
-CH3
-CH3
-CH3
-CH3
-CH3
-CH3
-CH3
-CH3
-H
-H
-H
-H
-H
-H
-H
-H
-H
-OMe
-OEt
200
580
260
920
300
1800
21
600
750
780
350
480
2900
570
2500
nd
nd
nd
nd
nd
1
nd
nd
49
2
0.8
nd
2
0.4
2
8
33
9
2
3
26
6
11
42
3
11
7
46
10
30
1800
7500
2600
1600
2300
9100
310
7400
850
450
2600
1000
11000
1200
11000
0.5
4
37-59
nd
nd
3-4
3
3
nd
9-13
4-6
2
nd
39-100
nd
nd
nd
-OnPr
-OnBu
0.3
0.3
2.8
4.4
<0.4
0.7
<0.5
<0.5
0.4
0.4
2
-OCH2CH(CH)2
-OCH2CH2OCH3
-OPh
-O-4-Pyr
-SCH3
-(CH3)2
-H
-OnBu
-H
-OnBu
-CH2CHdCH2
-CH2CHdCH2
-CH2CHdCH2
-CH2CH2CH3
-CH2CH2CH3
-OnPr
-H
-OnBu
-H
-OCH2CH2OCH3
-H
-OnBu
0.3
2
-H
-OCH2CH2OCH3
a
See Experimental Section for details of experimental assays. Standard deviations for enzyme assays were typically (60% of the
b
mean or less. Data obtained from both single and multiple determinations; for experimental details see ref 19. nd, not determined.
specific for this particular compound and those that
have similarly situated sulfonamide groups.
and 42 in Table 2, series 48-50 and series 58-60 in
Table 3, and series 61, 64, and 65 in Table 4. However,
compound 27, a pyridyl derivative of compound 23,
showed a greater absorption potential than would be
anticipated from its size alone. A recent report has
shown that several pyridyl-substituted compounds have
a greater transcellular permeability than the corre-
sponding phenyl-substituted molecule by an undeter-
mined mechanism.26 The addition of hydrophilicity in
the molecules also reduced its absorption potential as
exemplified by comparing compounds 44 vs 48 and 55
In Vitr o Absor p tion . The relative peroral intestinal
absorption potentials of select compounds shown in
Tables 1-4 were predicted from in vitro rat ileum
transport studies.19,25 The results suggest that there was
a general trend toward lower absorption when the
substitutions on the molecules became larger, i.e. in-
crease in molecular radius. Examples of this can be seen
in series 19 and 20, series 19, 21, and 22, and series 23
and 24 from Table 1, series 38 and 39 and series 41

New MMP Inhibitors from Functionalized Aminoprolines
J ournal of Medicinal Chemistry, 2000, Vol. 43, No. 26 4953
Ta ble 5. Pharmacokinetic and in Vivo Efficacy Data
time over
100 ng/mL (h)
in vivoa damage
reduction (%)
no.
p value
27
49
50
64
66
72
0
5
21*
26*
5
0
17*
0.43
0.03
0.01
0.32
0.50
0.05
4.5
3.5
0
0
4
a
Compounds were administered orally twice daily at a dose of
35 mg/kg. The data are expressed as the mean percent inhibition
of joint damage relative to a vehicle-treated control group. Each
group consisted of 15 rats. *p < 0.05. The broad-spectrum MMP
inhibitor CGS-27023 was used as a positive control in this model
and inhibited joint damage by 26 ( 6% (mean ( SD) from 13
separate experiments.
Ch a r t 1. Plasma Concentration vs Time Profile after
Cocktail Oral Administration of MMPIs
F igu r e 2. Surface structure of the stromelysin-24 complex.
single-compound experiment which served to validate
each data set and illuminate problems such as satura-
tion of elimination. The data suggests that most of the
compounds have somewhat short half-lives and low
absolute bioavailability. We were able to develop an
empirical relationship between plasma concentrations
and in vivo efficacy data. When plasma concentrations
were >100 ng/mL for >3 h as in compounds 49, 50, and
72, seven of eight compounds demonstrated in vivo
efficacy (data not shown). We were therefore able to use
the technique as a means of screening potent compounds
to narrow the list of candidates for efficacy screening
in more resource intensive in vivo studies.
The pharmacokinetics of compound 50 was investi-
gated in detail in rats as a single-compound study. It
was dosed intravenously and orally in male Sprague-
Dawley rats in a parallel group design. The data from
the study is depicted in Chart 2 and interpreted in Table
6 and supports the N-in-One data. Compound 50 dem-
onstrated an oral bioavailability of 6% with a half-life
of 1.7 h. The volume of distribution was relatively low
and the clearance was high. Despite this, the compound
was sufficiently available at the articular cartilage to
produce efficacy in vivo when dosed twice per day.
In Vivo Effica cy. The in vivo activity of selected
compounds was determined in rats using the iodoac-
etate-induced arthritis model,27-29 and the results are
summarized in Table 5. Compounds were administered
orally twice daily at a dose of 35 mg/kg for the first 7
days after intraarticular injection of iodoacetate. Ani-
mals were sacrificed 21 days after iodoacetate admin-
F igu r e 3. Binding interactions in the stromelysin-24 com-
plex.
vs 59. Several compounds (21, 22, 29, 55, and 56)
showed substantial partitioning into the intestinal
tissue based on significant disappearance of drug in the
donor (mucosal) chamber. In effect, it is possible that
these compounds may be absorbed to a greater extent
in vivo relative to that predicted from in vitro studies
and will depend, in part, on the compounds solubility/
protein binding in blood. Finally, there is evidence that
other compounds from this general class are substrates
for the various concentration-dependent efflux systems
(including P-glycoprotein), encoded by the MDR1 gene,
present in the enterocytes of the intestinal tract (data
not shown). Therefore, the extent of absorption will
depend on the total dose administered and the affinity
for the compound for the efflux transporter. Further
characterization would be necessary to better under-
stand potential solubility, tissue partitioning behavior,
and/or efflux mechanisms on overall peroral absorption.
P K a n d Effica cy An a lysis. The pharmacokinetics
for many of the aminopyrrolidine-based hydroxamates
were investigated using the N-in-One (cassette dosing)
technique, and the data for many of these is shown in
Table 5 and depicted in Chart 1. The studies were
conducted in fasted rats (n ) 3/4) using 4-10 hydrox-
amates per experiment at dose of 35/N mg/kg of each
hydroxamate. Each animal study included a control
compound that had been previously characterized in a

4954 J ournal of Medicinal Chemistry, 2000, Vol. 43, No. 26
Natchus et al.
phosphomolybdic acid in 95% ethanol. Final compounds were
typically purified either by flash chromatography on silica gel
(E. Merck, 40-63 mm) or by preparative reverse-phase high-
pressure liquid chromatography (RP-HPLC) using a Waters
model 4000 Delta Prep instrument equipped with a Waters
Symmetry preparative steel column (C-18, 19 m × 300 mm)
as the stationary phase. The mobile phase employed 0.1%
formic acid with acetonitrile as the organic modifier. Both
isocratic and linear gradient methods were used as appropriate
and the flow rate was 20 mL/min. Analytical purity was
assessed by RP-HPLC using a Waters 600 system equipped
with a diode array spectrometer (λ range 200-400 nm). The
stationary phase was a Waters Symmetry C-18 column (4.6
mm × 200 mm). The mobile phase employed 0.1% formic acid
with acetonitrile as the organic modifier and a flow rate of
1.0 mL/min. Analytical data is reported as retention time, tR,
in minutes (% acetonitrile, time, flow rate).
Ch a r t 2. Mean Plasma Concentration vs Time Profile
after Administration of Compound 50 to Rats
1H NMR spectra were recorded on a Varian Unity-300
instrument. Chemical shifts are reported in parts per million
(ppm, δ units). Coupling constants are reported in units of
hertz (Hz). Splitting patterns are designated as: s, singlet; d,
doublet; t, triplet; q, quartet; m, multiplet; br, broad. Low-
resolution mass spectra (MS) were recorded on a Micromass
Platform quadrupole mass spectrometer. Mass spectra were
acquired in either the positive or negative ion mode under
electrospray ionization (ESI). Combustion analyses were per-
formed internally.
Ta ble 6. Mean (%CV) PK Parameters for Compound 50 in
Rats
route
t1/2 (h)
CLt (mL/min/kg)
30 (12)
Vdss (mL/kg)
F (%)
iv
po
0.4 (22)
1.7 (41)
408 (23)
6
Human synovial proMMP-3 was obtained from Dr. Hideaki
Nagase, University of Kansas Medical Center, Kansas City,
KS. Human fibroblast proMMP-1, human MMP-9, and human
recombinant MMP-7 catalytic domain were obtained from Dr.
Howard Welgus, J ewish Hospital, St. Louis, MO. Human
recombinant MMP-8 catalytic domain was obtained from Dr.
Harald Tschesche, University Bielefeld, Bielefeld, Germany.
Human recombinant proMMP-2 was purified from CHO cells
as described below. Human recombinant truncated MMP-3
and truncated MMP-1 were purified from E. coli cells as
described below. Mca-Pro-Leu-Gly-Leu-Dpa-Ala-Arg-NH2 was
purchased from Bachem Bioscience, King of Prussia, PA.
istration, and joint damage was assessed. Compounds
49, 50, and 72 significantly (p e 0.05) inhibited joint
damage, whereas 27, 64, and 66 were not effective. All
of the compounds that significantly inhibited cartilage
degradation maintained plasma concentrations above
100 ng/mL for greater than 3 h, whereas all of the
compounds that were ineffective demonstrated much
lower plasma concentrations of drug.
Con clu sion
1N-(4-Met h oxyb en zen esu lfon yl)-2(R)-ca r b om et h oxy-
4(R)-m eth a n esu lfon oxyp yr r olid in e (2). The starting alco-
hol 8 (17.9 g, 57 mmol) was taken in CH2Cl2 (100 mL) and
Et3N (25 mL) was added. Methanesulfonyl chloride was added
dropwise and the resulting mixture was stirred overnight at
room temperature. The following the mixture was partitioned
between water and EtOAc. The organic layer was washed with
brine, dried over MgSO4, filtered and evaporated to give 24.3
g of solid product which was recrystallized from hexane:EtOAc
(∼1:1) to give 18.3 g of pure white solid. 1H NMR: (CDCl3,
300 MHz) δ 2.34 (ddd, J ) 14.5, 9.3, 5.1 Hz, 1H), 2.51-2.57
(m, 1H), 2.97, (s, 3H), 3.61 (ddd, J ) 11.9, 2.6, 1.1 Hz, 1H),
3.67 (dd, J ) 11.9, 4.8 Hz, 1H), 3.74 (s, 3H), 3.88 (s, 3H), 4.56
(dd, J ) 9.5, 6.8 Hz, 1H), 5.16 (m, 1H), 7.00 (ddd, J ) 9.0, 2.6,
2.6 Hz, 2H), 7.85 (ddd, J ) 9.1, 2.6, 2.6 Hz, 2H). CI+ MS: m/z
(rel intensity) 411 ([M + NH4]+, 25), 394 ([M + H]+, 21), 224
(82), 155 (23), 128 (100).
A diverse variety of aminopyrrolidine-based hydrox-
amate inhibitors are described here and are syntheti-
cally available in optically pure form from cis-hydroxy-
D-proline. They tend to be very potent for most members
in the MMP family with the exception of MMP-1 and
-7. This selectivity against the shallow S1′ pocket
enzymes can be enhanced by incorporating long-chain
aliphatic groups at the 4-position of the aromatic
sulfonamide group. X-ray crystallography data for a
stromelysin-24 complex confirms that this aromatic
sulfonamide structure indeed fits into the S1′ pocket
which helps explain the selectivity observations. The
inhibitors tended to show low to moderate absolute
bioavailability with short half-lives. The in vivo data for
this class of inhibitors seemed to be highly dependent
on the PK profile. Compounds such as 49 and 50 which
maintained blood levels above 100 ng/mL for 3 h or more
tended to show in vivo activity.
1N-(4-Met h oxyb en zen esu lfon yl)-2(R)-ca r b om et h oxy-
4(S)-a zid op yr r olid in e (3a ). The starting mesylate 2 (4.2 g,
10.7 mmol) was taken in 15 mL of dry DMF in the presence of
NaN3 (695 mg, 10.7 mmol). The resulting mixture was heated
to 55 °C for 26 h and then partitioned between water and
EtOAc. The organic layer was then washed with brine, dried
over MgSO4, filtered and evaporated. The resulting crude oil
was chromatographed over flash silica with hexane:EtOAc (5:1
to 3:1) to provide 2.87 g (79%) of pale yellow oil which solidified
Exp er im en ta l Section
All commercial chemicals and solvents are reagent grade
and used without further purification unless otherwise speci-
fied. The following solvents and reagents have been abbrevi-
ated: tetrahydrofuran (THF), ethyl ether (Et2O), dimethyl
sulfoxide (DMSO), ethyl acetate (EtOAc), dichloromethane
(DCM), trifluoroacetic acid (TFA), dimethylformamide (DMF),
methanol (MeOH). All reactions except those in aqueous media
were carried out with the use of standard techniques for the
exclusion of moisture. Reactions were monitored by thin-layer
chromatography on 0.25-mm silica gel plates (60F-254, E.
Merck) and visualized with UV light, iodine vapors, or 5%
1
upon standing. H NMR: (CDCl3, 300 MHz) δ 2.15-2.21 (m,
2H), 3.39 (ddd, J ) 11.2, 3.3, 0.9 Hz, 1H), 3.69 (dd, 11.2, 4.9
Hz, 1H), 3.77 (s, 3H), 3.88 (s, 3H), 4.17-4.23 (m, 1H), 4.27
(ddd, J ) 7.3, 7.3, 0.6 Hz, 1H), 7.00 (ddd, 9.0, 2.6, 2.6 Hz, 2H),
7.81 (ddd, J ) 9.0, 2.6, 2.6 Hz, 2H). CI+ MS: m/z (rel intensity)
358 ([M + NH4]+ 50), 341 ([M + H]+, 67), 315 (95), 145 (100).
1N-(4-Met h oxyb en zen esu lfon yl)-2(R)-ca r b om et h oxy-
4(S)-a m in op yr r olid in e (3). The starting azide 3a (1.18 g,
3.48 mmol) was taken in 100 mL of EtOH:THF:HCO2H (5:1:

New MMP Inhibitors from Functionalized Aminoprolines
J ournal of Medicinal Chemistry, 2000, Vol. 43, No. 26 4955
0.1) and hydrogenated at room temperature under 54 psi of
hydrogen in the presence of 100 mg of 10% Pd-C for 16 h.
The mixture was then filtered through a pad of Celite,
concentrated to an oil and recrystallized from hexane:EtOAc
to give 720 mg (58%) of the desired product as the formate
salt. 1H NMR: (CDCl3, 300 MHz) δ 1.72-1.81 (m, 1H), 2.00
(ddd, J ) 13.0, 5.7, 5.7 Hz, 1H), 2.86 (dd, J ) 9.7, 5.3 Hz, 1H),
3.46-3.57 (m, 2H), 3.57 (s, 3H), 3.73 (s, 3H), 4.21 (dd, J ) 8.2,
6.2 Hz, 1H), 6.86 (br d, J ) 9.0 Hz, 2H), 7.66 (br d, J ) 9.0 Hz,
2H). CI+ MS: m/z (rel intensity) 315 ([M + H]+, 12), 177 (13),
143 (42), 123 (60), 109 (100).
Na2CO3. The organic layer was washed with brine, dried over
MgSO4, filtered and evaporated. The crude oil was chromato-
graphed over flash silica with EtOAc:MeOH (20:1 to 4:1) to
give 11.5 g (91%) of the title amine. 1H NMR: (CDCl3, 300
MHz) δ 1.93 (ddd, J ) 12.8, 8.6, 6.6, 1H), 2.16 (ddd, J ) 12.8,
5.6, 5.6 Hz, 1H), 2.29 (s, 3H), 3.09 (dd, J ) 9.8, 5.3 Hz, 1H),
3.30-3.40 (m, 1H), 3.62 (dd, J ) 9.8, 5.6 Hz, 1H), 3.74 (s, 3H),
3.88 (s, 3H), 4.34 (dd, J ) 8.4, 5.4 Hz, 1H), 6.99 (ddd, J ) 8.9,
3.1, 2.1 Hz, 2H), 7.82 (ddd, J ) 9.0, 2.9, 2.2 Hz, 2H). CI+ MS:
m/z (rel intensity) 423.5 ([M + NH4]+, 42), 406.5 ([M + H]+,
100).
1N-(4-Met h oxyb en zen esu lfon yl)-2(R)-ca r b om et h oxy-
4(S)-p r op yla m in op yr r olid in e (4). The starting amine 3 (810
mg, 2.6 mmol) was dissolved in 8 mL of methanol and stirred
for 48 h in the presence of propianaldehyde (206 mL, 2.86
mmol), sodium cyanoborohydride (180 mg, 2.86 mmol), sodium
acetate (810, 9.9 mmol) and 25 drops of acetic acid. The
mixture was evaporated to dryness and then partitioned
between dilute NaHCO3 and EtOAc and the organic layer
washed 2× with NaHCO3, 1× with brine, dried over MgSO4,
filtered and evaporated. 1H NMR: (CDCl3, 300 MHz) δ 0.87
(t, J ) 7.4 Hz, 3H), 1.49-1.64 (m, 2H), 2.10-2.22 (m, 1H),
2.31 (ddd, J ) 13.2, 7.0, 2.7 Hz, 1H), 2.69-2.80 (m, 2H), 3.28
(dd, J ) 10.1, 6.8 Hz, 1H), 3.55 (s, 3M), 3.63-3.86 (m, 2H),
3.76 (s, 3H), 4.35 (dd, J ) 9.2, 2.9 Hz, 1H), 6.92 (br d, J ) 9.1
Hz, 2H), 7.65 (br d, J ) 9.2 Hz, 2H). ESI MS: m/z (rel
intensity) 357.3 ([M + H]+, 100).
1N-(4-Met h oxyb en zen esu lfon yl)-2(R)-ca r b om et h oxy-
4(S)-N-m eth yl-N-m eth a n esu lfon yla m in op yr r olid in e (6).
The title compound was prepared (284 mg, 46%) from amine
6b (502 mg, 1.47 mmol) as described for compound 5. 1H
NMR: (CDCl3, 300 MHz) δ 2.15-2.33 (m, 2H), 2.74 (s, 3H),
2.82 (s, 3H), 3.33 (dd, J ) 9.5, 7.5 Hz, 1H), 3.60 (dd, J ) 9.5,
8.2 Hz, 1H), 3.67 (s, 3H), 3.90 (s, 3H), 4.53 (dd, J ) 7.9, 3.5
Hz, 1H), 4.64-4.78 (m, 1H), 7.02 (br d, J ) 9.0 Hz, 2H), 7.81
(br d, J ) 9.0 Hz, 2H). CI+ MS: m/z (rel intensity) 423.5 ([M
+ NH4]+, 42), 406.5 ([M + H]+, 100).
1N-(4-Met h oxyb en zen esu lfon yl)-2(R)-ca r b om et h oxy-
4(S)-N-(m eth an esu lfon ylpr opyl)am in opyr r olidin e (7). The
starting amine 4 (783 mg, 2.20 mmol) was converted to the
title compound (624 mg, 66%) as described for 7. 1H NMR:
(CDCl3, 300 MHz) δ 0.86 (t, J ) 7.4 Hz, 3H), 1.51-1.66 (m,
2H), 2.16-2.23 (m, 2H), 2.82 (s, 3H), 2.91 (ddd, J ) 13.9, 9.5,
6.0 Hz, 1H), 3.01 (ddd, J ) 14.1, 8.1, 4.4 Hz, 1H), 3.24 (dd, J
) 8.8, 8.8 Hz, 1H), 3.64 (dd, J ) 9.0, 8.2 Hz, 1H), 3.68 (s, 3H),
3.88 (s, 3H), 4.47-4.58 (m, 2H), 7.00 (ddd, J ) 9.0, 2.6, 2.6
Hz, 2H), 7.80 (ddd, J ) 9.0, 2.6, 2.6 Hz, 2H). ESI MS: m/z (rel
intensity) 452 ([M + NH4]+, 25), 435 ([M + H]+, 75), 265 (100),
155 (75), 126 (40).
1N-(4-Met h oxyb en zen esu lfon yl)-2(R)-ca r b om et h oxy-
4(R)-h yd r oxyp yr r olid in e (8). cis-Hydroxy-D-proline (1; 50
g, 0.38 mol) was dissolved in water:dioxane (1:1, 300 mL) with
triethylamine (135 mL, 0.96 mol). 4-Methoxyphenylsulfonyl
chloride (87 g, 0.42 mol) was added along with 2,6-(dimethy-
lamino)pyridine (4.6 g, 0.038 mol) and the mixture was stirred
14 h at room temperature. The mixture was then concentrated
and diluted with EtOAc. Layers were separated and the
organic layer was washed 2× with 1 N HCl, 1× with brine,
dried over MgSO4, filtered and evaporated to give 83 g of solid
material which was dissolved in MeOH (500 mL). Thionyl
chloride (50 mL) was added dropwise and the resulting
mixture stirred for 14 h. The mixture was then evaporated to
dryness and triturated with CHCl3 to give 85 g (69%) of white
solid which was sufficeintly pure to carry forward without
purification. 1H NMR: (CDCl3, 300 MHz) δ 2.07 (ddd, J ) 14.1,
3.8, 1.6 Hz, 1H), 2.16 (ddd, J ) 14.1, 9.5, 4.4 Hz, 1H), 3.31
(dd, J ) 10.3, 4.2 Hz, 1H), 3.48-3.54 (m, 2H), 3.76 (s, 3H),
3.89 (s, 3H), 4.28-4.34 (m, 2H), 6.98 (ddd, J ) 9.0, 2.5, 2.5
Hz, 2H), 7.79 (ddd, J ) 9.1, 2.6, 2.6 Hz, 2H). CI+ MS: m/z (rel
intensity) 316 ([M + H]+, 100), 256 (300), 146 (45), 114 (20).
1N-(4-Met h oxyb en zen esu lfon yl)-2(R)-ca r b om et h oxy-
4(S)-(3N-m eth ylh yd a n to-1N-yl)p yr r olid in e (9). Diethyl
azodicarboxylate (1.8 mL, 11.42 mmol) was added to a stirred
solution of the starting alcohol 8 (3.0 g, 9.51 mmol), triph-
enylphosphene (3.74 g, 9.51 mmol), and 1-methylhydantoin
(1.3 g, 11.42 mmol) in 30 mL of CH2Cl2 and stirred for 16 h at
room temperature. The mixture was then chromatographed
over flash silica with hexane and then hexane:EtOAc (1:1) to
give a colorless glass which was recrystallized from methanol
to give 3.2 g (82%) of white powder. 1H NMR: (CDCl3, 300
MHz) δ 2.15 (ddd, J ) 12.8, 7.5, 2.9 Hz, 1H), 2.83-2.91 (m,
1H), 2.97 (s, 3H), 3.69 (d, J ) 8.06 Hz, 2H), 3.77 (s, 3H), 3.79
(s, 2H), 3.90 (s, 3H), 4.50 (dd, J ) 9.3, 2.9 Hz, 1H), 4.78-4.89
(m, 1H), 7.02 (ddd, J ) 9.2, 2.6 Hz, 2H), 7.83 (ddd, J ) 9.2,
2.6 Hz, 2H). ESI MS: m/z (rel intensity) 412.1 ([M + H]+, 100),
429.1 ([M + NH4]+, 45).
1N-(4-Met h oxyb en zen esu lfon yl)-2(R)-ca r b om et h oxy-
4(S)-m eth a n esu lfon yla m in op yr r olid in e (5). The primary
amine 3 (502 mg, 1.60 mmol) was taken in 5 mL of methylene
chloride and 0.5 mL of triethylamine and treated with meth-
anesulfonyl chloride (200 µL, 2.58 mmol) via syringe. The
mixture was stirred for 2 h and then partitioned between 1 N
HCl and EtOAc. The organic layer was washed with brine,
dried over MgSO4, filtered and evaporated to give 684 mg of
crude material which was chromatographed over flash silica
with hexane:EtOAc (2:1 to 1:1) to give 104 mg (14%) of
disulfonylated material 5a and 393 mg (61%) of monosulfo-
1
nylated material 5. H NMR: (CDCl3, 300 MHz) δ 2.21 (ddd,
J ) 13.4, 8.2, 6.4 Hz, 1H), 2.96 (s, 3H), 3.33 (dd, J ) 10.4, 4.9
Hz, 1H), 3.65 (dd, J ) 10.4, 5.9, Hz, 1H), 3.71 (s, 3H), 3.88 (s,
3H), 4.10-4.20 (m, 1H), 4.49 (dd, J ) 8.4, 5.5 Hz, 1H), 4.74
(d, J ) 7.9 Hz, 1H), 7.00 (ddd, J ) 9.0, 2.6, 2.6 Hz, 2H), 7.82
(ddd, J ) 9.0, 2.5, 2.5 Hz, 2H). CI+ MS: m/z (rel intensity)
410 ([M + NH4]+, 15), 393 ([M + H]+, 10), 203 (100).
1N-(4-Met h oxyb en zen esu lfon yl)-2(R)-ca r b om et h oxy-
4(S)-N-bis(m eth a n esu lfon yl)a m in op yr r olid in e (5a ). 1H
NMR: (CDCl3, 300 MHz) δ 2.23-2.31 (m, 1H), 2.74-2.86 (m,
1H), 3.28 (s, 6H), 3.68 (dd, J ) 10.4 Hz, 1H), 3.74 (s, 3H), 3.81
(dd, J ) 10.5 Hz, 1H), 3.98 (s, 3H), 4.38 (dd, J ) 11.4, 4.2 Hz,
1H), 5.08-5.21 (m, 1H), 7.01 (br d, J ) 8.9 Hz, 2H), 7.81 (br
d, J ) 8.9 Hz, 2H). ESI MS: m/z (rel intensity) 488.3 ([M +
NH4]+, 15), 471.3 ([M + H]+, 10).
1N-(4-Met h oxyb en zen esu lfon yl)-2(R)-ca r b om et h oxy-
4(S)-N-for m yla m in op yr r olid in e (6a ). The free amine 3
(20.2 g, 64 mmol) was converted to 17.5 g (80%) of the title
formamide as described by Krishnamurthy.30 1H NMR: (CDCl3,
300 MHz) δ 2.20-2.38 (m, 2H), 3.42 (dd, J ) 10.6, 2.9 Hz,
1H), 3.55 (dd, J ) 11.0, 5.5 Hz, 1H), 3.71 (s, 3H), 3.89 (s, 3H),
4.52-4.62 (m, 2H), 6.19 (d, J ) 6.6 Hz, 1H), 7.01 (ddd, J )
9.0, 2.4, 2.0 Hz, 2H), 7.85 (ddd, J ) 9.0, 3.1, 2.0 Hz, 2H), 8.03
(s, 1H). ESI: m/z (rel intensity) 360.1 ([M + NH3]+, 68), 343.1
([M + H]+, 100).
1N-(4-Met h oxyb en zen esu lfon yl)-2(R)-ca r b om et h oxy-
4(S)-N-m eth yla m in op yr r olid in e (6b). The formamide 6a
(13.3 g, 342 mmol) was taken in 20 mL of THF and cooled to
0 °C and a solution of BH3‚THF (85.8 mL, 1 M in THF, 85.5
mmol) was added via syringe. After 10 min, the solution was
heated to 60 °C for 2 h and then cooled to room temperature
and quenched with 20 mL of MeOH. An additional 150 mL of
MeOH was then added which had been pretreated with 15 mL
of SOCl2 and the resulting mixture was heated to 60 °C for 3
h. The resulting solution was then condensed to an oil by
rotary evaporation and partitioned between EtOAc and dilute
1N-(4-Met h oxyb en zen esu lfon yl)-2(R)-ca r b om et h oxy-
4(S)-(m or ph olin -1N-yl)pyr r olidin e (10). The starting amine
3 (590 mg, 1.88 mmol) was taken in 4 mL of DMF and 1 mL
of NEt3 and treated with 1 mL of 2-bromoethyl ether. The

4956 J ournal of Medicinal Chemistry, 2000, Vol. 43, No. 26
Natchus et al.
resulting mixture was then heated to 60 °C for 3 h and
partitioned between dilute NaCO3 and EtOAc. The organic
layer was then dried over MgSO4, filtered and evaporated. The
crude residue was chromatographed over flash silica with
EtOAc:MeOH (9:1) to give 348 mg (48%) of the title compound
as a white solid. 1H NMR: (CDCl3, 300 MHz) δ 1.97 (ddd, J )
12.5, 8.9, 8.9 Hz, 1H), 2.18 (ddd, J ) 12.5, 5.7, 2.4 Hz, 1H),
2.34-2.49 (m, 4H), 3.01-3.12 (m, 2H), 3.66 (dd, J ) 4.8, 4.8
Hz, 4H), 3.69-3.74 (m, 1H), 3.77 (s, 3H), 3.90 (s, 3H), 4.35
(dd, J ) 9.7, 2.4 Hz, 1H), 7.01 (ddd, J ) 8.8, 2.6, 2.6 Hz, 2H),
7.82 (ddd, J ) 8.8, 2.5, 2.5 Hz, 2H). ESI MS: m/z (rel intensity)
385.1 ([M + H]+, 100).
1N-(4-Met h oxyb en zen esu lfon yl)-2(R)-ca r b om et h oxy-
4(S)-(γ-ch lor osu lfon am ido)pyr r olidin e (11). The free amine
3 (7.51 g, 23.8 mmol) was taken in 15 mL of methylene chloride
in the presence of triethylamine and treated with 3-chloro-
propylsulfonyl chloride (3.76 mL, 31.0 mmol) at room temper-
ature. The resulting mixture was allowed to stir for 18 h and
then it was diluted with 200 mL of methylene chloride, washed
with 1 N HCl, dried over MgSO4, filtered and evaporated. The
crude material was adsorbed onto silica and then eluted
through a column of flash silica with hexane:EtOAc (2:1 to
1:3) to give 7.21 g (67%). 1H NMR: (CDCl3, 300 MHz) δ 2.20-
2.38 (m, 4H), 3.20 (dd, J ) 7.4, 7.4 Hz, 2H), 3.38 (dd, J ) 10.4,
4.6 Hz, 1H), 3.63-3.72 (m, 2H), 3.72 (s, 3H), 3.90 (s, 3H), 4.11-
4.22 (m, 1H), 4.53 (dd, J ) 8.4, 5.7 Hz, 1H), 4.72 (d, J ) 8.1
Hz, 1H), 7.03 (ddd, J ) 8.8, 2.9, 2.0 Hz, 2H), 7.85 (ddd, J )
8.9, 2.9, 2.0 Hz, 2H). ESI MS: m/z (rel intensity) 455.0 ([M +
H]+, 100), 419.0 ([M - Cl]+, 90).
converted and purified as described for compound 3a to give
the title azide (34.5 g, 59%) as a yellow gum. This compound
appears in the 1H NMR spectrum as a distinct pair of rotomers.
1H NMR: (CDCl3, 300 MHz) δ [1.43 (s), 1.48 (s)] 9H, [2.17 (dd,
J ) 6.5, 6.5 Hz), 2.21 (dd, 6.4, 6.4 Hz)] 1H, 2.26-2.41 (m, 1H),
[3.49 (dd, J ) 11.4, 3.5 Hz), 3.60 (dd, J ) 11.5, 2.6 Hz)] 1H,
3.70-3.74 (m, 1H), 3.76 (br s, 3H), 4.17-4.24 (m, 1H), [4.35
(dd, J ) 7.6, 7.6 Hz), 4.43 (dd, J ) 7.1, 7.1 Hz) 1H.
1N-ter t-Bu toxyca r bon yl-2(R)-ca r bom eth oxy-4(S)-a m i-
n op yr r olid in e (14). The azide 14b (26 g, 101 mmol) was
hydrogenated and purified as described for compound 3 to give
the title amine (15.7 g, 61%) as a pale yellow gum. This
compound appears in the 1H NMR spectrum as a distinct pair
1
of rotomers. H NMR: (CDCl3, 300 MHz) δ [1.35 (s), 1.41 (s)]
9H, 1.75-2.02 (m, 2H), 2.93-3.06 (m, 1H), 3.38-3.50 (m, 2H),
[3.62 (s), 1.64 (s)] 3H, 4.20-4.28 (m, 1H). ESI MS: m/z (rel
intensity) 245.0 ([M + H]+, 55), 144.8 ([M - Boc + H]+, 100).
1N-ter t-Bu toxyca r bon yl-2(R)-ca r bom eth oxy-4(S)-(p yr -
r olid in -1N-yl)p yr r olid in e (15). The starting free amine 14
(7.51 g, 30.7 mmol) was treated with 8 mL of 1,4-dibromobu-
tane to give the title pyrrolidine (6.85 g, 75%) as described for
1
compound 10. This compound appears in the H NMR spec-
trum as a distinct pair of rotomers. 1H NMR: (CDCl3, 300
MHz) δ [1.40 (s), 1.46 (s)] 9H, 1.77-1.89 (m, 4H), 2.10-2.34
(m, 2H), 2.50-2.66 (m, 4H), 2.88-3.05 (m, 1H), 3.25-3.37 (m,
1H), 3.70-3.86 (m, 1H), 3.74 (s, 3H), [4.33 (dd, J ) 9.0, 2.2
Hz), 4.43 (dd, J ) 8.7, 1.8 Hz)] 1H. ESI MS: m/z (rel intensity)
299.0 ([M + H]+, 100), 242.9 (50), 198.9 ([M - Boc + H]+, 5).
1N-(4-P h en oxyben zen esu lfon yl)-2(R)-ca r b om et h oxy-
4(S)-(p yr r olid in -1N-yl)p yr r olid in e (16). The starting pro-
tected amine 15 (1.36 g, 4.56 mmol) was taken in 30 mL of
CH2Cl2 and treated with 2 mL of trifluoroacetic acid. The
resulting mixture was allowed to stir for 2 h, evaporated to
dryness and tritutated with CHCl3. The residue was then
taken in 30 mL of dioxane:water:Et3N (3:1:2) and treated with
4-phenoxybenzenesulfonyl chloride.31 The resulting mixture
was stirred for 16 h and then partitioned between EtOAc and
dilute NaHCO3. The organic layer was dried over MgSO4,
filtered and evaporated. The crude residue was adsorbed onto
silica and eluted through a column of flash silica to give 884
mg (45%) of clear gum which solidified upon standing. 1H
NMR: (CDCl3, 300 MHz) δ 1.69-1.83 (m, 4H), 2.05 (ddd, J )
12.6, 9.3, 9.3 Hz, 1H), 2.17 (ddd, J ) 12.8, 6.0, 3.1 Hz, 1H),
2.38-2.54 (m, 4H), 2.93-3.05 (m, 1H), 3.08 (dd, J ) 8.2, 8.2
Hz, 1H), 3.72 (dd, J ) 8.2, 6.0 Hz, 1H), 3.78 (s, 3H), 4.33 (dd,
J ) 9.3, 2.9 Hz, 1H), 7.05 (ddd, J ) 8.7, 2.9, 1.8 Hz, 2H), 7.08-
7.13 (m, 2H), 7.22-7.29 (m, 1H), 7.40-7.48 (m, 2H), 7.82 (ddd,
J ) 8.6, 2.9, 1.8 Hz, 2H). ESI MS: m/z (rel intensity) 431.0
([M + H]+, 100).
1N-(4-Met h oxyb en zen esu lfon yl)-2(R)-ca r b om et h oxy-
4(S)-(γ-su lta m -1N-yl)p yr r olid in e (12). The substrate 11 was
taken in 20 mL of DMF and treated at room temperature with
1,8-diazabicyclo[5.4.0]undec-7-ene (3 mL, 20.1 mmol). The
resulting mixture was stirred for 18 h and then diluted with
hexanes:EtOAc (3:1) and washed twice with 1 N HCl, once
with brine, dried over MgSO4, filtered and evaporated. The
residue was adsorbed onto silica and eluted through a column
of flash silica with hexane:EtOAc (1:1 to 1:3) to give 1.28 g
(77%). 1H NMR: (CDCl3, 300 MHz) δ 2.26-2.45 (m, 4H), 3.12-
3.27 (m, 4H), 3.41 (dd, J ) 9.9, 6.6 Hz, 1H), 3.67 (dd, J ) 9.7,
6.8 Hz, 1H), 3.70 (s, 3H), 3.89 (s, 3H), 4.07-4.17 (m, 1H), 4.51
(dd, J ) 8.6, 4.0 Hz, 1H), 7.01 (ddd, J ) 9.0, 2.9, 2.0 Hz, 2H),
7.82 (ddd, J ) 9.0, 2.9, 2.1 Hz, 2H). ESI MS: m/z (rel intensity)
419.0 ([M + H]+, 100).
1N-ter t-Bu t oxyca r b on yl-2(R)-ca r b om et h oxy-4(R)-h y-
d r oxyp yr r olid in e (13). cis-4-Hydroxy-D-proline methyl ester
hydrochloride (40 g, 221 mmol) was taken in 250 mL of water:
acetone (2:3) in the presence of 60 mL of triethylamine and 1
g of 4-N,N-dimethylaminopyridine. Di-tert-butyl dicarbonate
(66 g, 303 mmol) was added slowly in portions and the
resulting mixture was allowed to stir for 16 h and then
concentrated to one-half of its original volume and then
partitioned between EtOAc and dilute NaH2PO4. The organic
layer was washed once with dilute NaH2PO4, once with brine,
dried over MgSO4, filtered and evaporated to give 58 g of
material (107%) which was carried forward withput purifica-
tion. This compound appears in the 1H NMR spectrum as a
1N-ter t-Bu t oxyca r b on yl-2(R)-ca r b om et h oxy-4(S)-(γ-
ch lor osu lfon a m id o)p yr r olid in e (17a ). The starting amine
14 (4.01 g, 16.4 mmol) was converted to 6.18 g (98%) of the
title sulfonamide as a white solid as described for compound
11. This compound appears in the 1H NMR spectrum as a
1
distinct pair of rotomers. H NMR: (CDCl3, 300 MHz) δ [1.43
(s), 1.48 (s)] 9H, 2.24-2.36 (m, 4H), 3.24 (dd, J ) 7.3, 7.7 Hz,
2H), [3.32 (dd, J ) 11.0, 5.9 Hz), 3.42 (dd, J ) 10.8, 4.8 Hz)]
1H, 3.70 (dd, J ) 6.1, 6.1 Hz, 2H), 3.77 (s, 3H), 3.82-3.92 (m,
1H), 4.08-4.39 (m, 1H), [4.35 (dd, J ) 6.6, 7.0 Hz), 4.43 (dd,
J ) 8.1, 4.2 Hz)] 1H, [4.92 (d, J ) 8.4), 5.01 (d, J ) 7.7 Hz)]
1H. ESI MS: m/z (rel intensity) 285.0 ([M - Boc + H]+, 100).
1
distinct pair of rotomers. H NMR: (CDCl3, 300 MHz) δ [1.44
(s), 1.48 (s)] 9H, 2.06-2.16 (m, 1H), 2.27-2.42 (m, 1H), 3.50-
3.75 (m, 2H), [3.80 (s), 3.81 (s)] 1H, 4.29-4.42 (m, 2H). ESI
MS: m/z (rel intensity) 262.9 ([M + NH4]+, 45), 245.9 ([M +
H]+, 100), 145.8 ([M - Boc + H]+, 90).
1N-ter t-Bu t oxyca r b on yl-2(R)-ca r b om et h oxy-4(S)-(γ-
su lta m -1N-yl)p yr r olid in e (17). The starting sulfonamide
17a (6.15 g, 16.0 mmol) was converted as described for
compound 12 to give 4.21 g (76%) of the title compound as a
white solid. This compound appears in the 1H NMR spectrum
1N-ter t-Bu toxycar bon yl-2(R)-car bom eth oxy-4(R)-m eth -
a n esu lfon ylp yr r olid in e (14a ). The starting alcohol 13 (58
g, 237 mmol) was converted to the title compound as described
for compound 2. The crude oil was filtered through a plug of
silica with hexane:EtOAc (1:1) and concentrated to give 73 g
(95%) of pale orange oil. This compound appears in the 1H
NMR spectrum as a distinct pair of rotomers. 1H NMR: (CDCl3,
300 MHz) δ [1.44 (s), 1.49 (s)] 9H, 2.48-2.60 (m, 2H), 3.03 (s,
3H), 3.75-3.84 (m, 1H), 3.77 (s, 3H), [4.42 (dd, J ) 5.9 Hz),
4.53 (dd, J ) 7.7, 4.0 Hz)] 1H, 5.21-5.29 (m, 1H).
1
as a distinct pair of rotomers. H NMR: (CDCl3, 300 MHz) δ
[1.40 (s), 1.46 (s)] 9H, 2.23-2.45 (m, 3H), 2.54 (ddd, J ) 13.5,
8.5, 8.5 Hz, 1H), 3.10-3.34 (m, 4H), 3.42-3.53 (m, 1H), 3.74
(s, 3H), 3.78-3.88 (m, 1H), 3.98-4.15 (m, 1H), [4.36 (dd, J )
8.8, 3.7 Hz), 4.43 (dd, J ) 9.2, 3.3 Hz)] 1H. ESI MS: m/z (rel
intensity) 249.0 ([M - Boc + H]+, 100).
1N-ter t-Bu toxyca r bon yl-2(R)-ca r bom eth oxy-4(S)-a zi-
d op yr r olid in e (14b). The mesylate 14a (73 g, 226 mmol) was
1N-(4-n -P r op oxyben zen esu lfon yl)-2(R)-ca r bom eth oxy-
4(S)-(γ-su lta m -1N-yl)p yr r olid in e (18). The starting car-

New MMP Inhibitors from Functionalized Aminoprolines
J ournal of Medicinal Chemistry, 2000, Vol. 43, No. 26 4957
bamate 17 (1.27 g, 3.67 mmol) was deprotected and sulfonyl-
ated as described for compound 16 to give 934 mg (57%) of
the title sultam as a solid. 1H NMR: (CDCl3, 300 MHz) δ 1.07
(t, J ) 7.4 Hz, 3H), 1.80-1.93 (m, 2H), 2.27-2.46 (m, 4H),
3.12-3.30 (m, 4H), 3.41 (dd, J ) 9.9, 6.8 Hz, 1H), 3.68 (dd, J
) 9.7, 6.8 Hz, 1H), 3.70 (s, 3H), 4.08-4.18 m, 1H), 4.51 (dd, J
) 8.6, 3.8 Hz, 1H), 7.00 (ddd, J ) 9.0, 2.7, 2.0 Hz, 2H), 7.81
(ddd, J ) 9.0, 2.7, 2.0 Hz, 2H). ESI MS: m/z (rel intensity)
464 ([M + NH4]+, 20).
(s, 3H), 3.65 (dd, J ) 9.2, 7.1 Hz, 1H), 3.87 (s, 3H), 3.98 (dd, J
) 9.0, 2.2 Hz, 1H), 4.01-4.14 (m, 1H), 7.16 (br d, J ) 8.61,
2H), 7.23 (d, J ) 6.8 Hz, 1H), 7.79 (d, J ) 8.8 Hz, 2H), 9.04 (s,
1H), 10.75 (1H). ESI MS: m/z (rel intensity) 394.1 ([M + H]+,
100). Analysis: C, H, N for C13H19N3O7S2‚0.3H2O.
1N-(4-n -Bu toxyph en ylsu lfon yl)-2(R)-N-h ydr oxycar box-
a m id o-4(S)-m eth a n esu lfon yla m in op yr r olid in e (24). Com-
pound 24 was prepared as described for compound 23 to give
1
a white solid. H NMR: (DMSO-d6, 300 MHz) δ 0.95 (t, J )
7.4, 3H), 1.39-1.42 (m, 2H), 1.61-1.79 (m, 3H), 1.98-2.06 (m,
1H), 2.82-2.94 (m, 1H), 2.89 (s, 3H), 3.64 (dd, J ) 9.0, 6.8 Hz,
1H), 3.99 (dd, J ) 9.2, 2.2 Hz, 1H), 4.02-4.14 (m, 3H), 7.14
(br d, J ) 9.0 Hz, 2H), 7.77 (br d, J ) 9.0 Hz, 2H). ESI MS:
m/z (rel intensity) 453.08 ([M + NH4]+, 50), 436.05 ([M + H]+,
100). Analysis: C, H, N for C16H25N3O7S2‚0.7H2O.
1N-(4-Met h oxyb en zen esu lfon yl)-2(R)-N-h yd r oxyca r -
boxa m id o-4(S)-a m in op yr r olid in e (19). The starting ester
3 (500 mg, 1.59 mmol), was taken in 5 mL of MeOH, treated
with NH2OK (1.92 mL, 0.86 M in methanol; solution prepared
as described in Fieser and Fieser, Vol 1, p 478) and stirred
overnight. The following morning, dry silica (1.5 mL) was
added to the mixture and the solvent removed under vacuum.
The dry silica was poured on the top of a flash silica gel column
which was subsequently eluted with EtOAc:MeOH (4:1 to 3:2)
to give 284 mg (56%) of white solid. 1H NMR: (CDCl3, 300
MHz) δ 1.49 (ddd, J ) 12.1, 7.9, 7.9 Hz, 1H), 1.85 (ddd, J )
12.3, 4.9, 4.9 Hz, 1H), 2.68 (dd, J ) 8.6, 5.9 Hz, 1H), 3.41-
3.57 (m, 2H), 3.87 (s, 3H), 3.93 (dd, J ) 8.4, 4.4 Hz, 1H), 7.14
(d, J ) 8.8 Hz, 2H), 7.80 (d, J ) 8.8 Hz, 2H). ESI MS: m/z (rel
intensity) 316.1 ([M + H]+, 100). Analysis: C, H, N for
1N-(4-P yr idin -4-yloxyben zen esu lfon yl)-2(R)-N-h ydr oxy-
ca r boxa m id o-4(S)-a m in op yr r olid in e (25). The title com-
pound was prepared as described for compound 23 to give a
1
white solid. H NMR: (DMSO-d6, 300 MHz) δ 1.81-1.95 (m,
1H), 2.07 (ddd, J ) 12.9, 6.3, 6.3 Hz, 1H), 2.99 (dd, J ) 9.7,
4.8 Hz, 1H), 3.53 (dd, J ) 9.7, 5.5 Hz, 1H), 3.63 (s, 3H), 4.34
(dd, J ) 5.7 8.2 Hz, 1H), 6.80 (m, 2H), 7.12 (d, J ) 8.8 Hz,
2H), 7.85 (d, J ) 8.8 Hz, 2H), 8.43-8.46 (m, 2H). ESI MS:
m/z (rel intensity) 378.1 ([M + H]+, 100). Analysis: C17H20
N4O7S2‚HCl‚0.3H2O.
-
C
12H17N3O5S‚0.8H2O.
1N-(4-n -Bu toxyben zen esu lfon yl)-2(R)-h yd r oxyca r box-
1N-(4-Meth oxyben zen esu lfon yl)-2(R)-h yd r oxyca r box-
a m id o-4(S)-N-m eth yl-N-m eth a n esu lfon yla m in op yr r oli-
d in e (26). Compound 24 was prepared as described for
a m id o-4(S)-a m in op yr r olid in e (20). The title compound was
prepared as described for compound 19 and then purified
further by recrystallizing from water. 1H NMR: (DMSO-d6,
300 MHz) δ 0.95 (td, J ) 7.1, 1.8 Hz, 3H), 1.47 (septet, J )
7.7 Hz, 2H), 1.69-1.88 (m, 4H), 2.67 (dd, J ) 7.3 Hz, 1H), 3.48
(ddd, J ) 13.6, 13.6, 5.5 Hz, 1H), 3.92 (dd, J ) 7.5, 3.3 Hz,
1H), 4.08 (t, J ) 6.3 Hz, 2H), 7.13 (br d, J ) 8.6 Hz, 2H), 7.77
(br d, J ) 8.6 Hz, 2H). ESI MS: m/z (rel intensity) 358 ([M +
H]+, 100). Analysis: C, H, N for C15H23N3O5S‚0.2H2O.
1
compound 23 to give a white solid. H NMR: (DMSO-d6, 300
MHz) δ 1.81-1.91 (m, 2H), 2.46 (s, 3H), 2.88 (s, 3H), 3.03 (dd,
J ) 9.4, 8.8 Hz, 1H), 3.57 (dd, J ) 9.7, 7.7 Hz, 1H), 3.86 (s,
3H), 4.07 (dd, J ) 6.4, 4.4 Hz, 1H), 4.58-4.71 (m, 1H), 7.16
(br d, J ) 9.0 Hz, 2H), 7.82 (br d, J ) 8.8 Hz, 1H). ESI MS:
m/z (rel intensity) 429.3 ([M + Na]+, 15), 425.0 ([M + NH4]+
20), 407.4 ([M + H]+, 100). Analysis: C, H, N for C14H21N3O7S2‚
0.4H2O.
1N-(4-Met h oxyb en zen esu lfon yl)-2(R)-N-h yd r oxyca r -
boxa m id o-4(S)-N-p r op yla m in op yr r olid in e (21). The start-
ing methyl ester 4 (11.3, g, 31.7 mmol) was taken in 30 mL of
methanol, treated with NH2OK (38 mL, 1.25 M in methanol;
solution prepared as described in Fieser and Fieser, Vol 1, p
478) and stirred for 16 h. The following morning, dry silica
(30 mL) was added to the mixture and the solvent removed
under vaccuum. The dry silica was poured on the top of a flash
silica gel column which was subsequently eluted with chloro-
form:methanol (8:2) to give a pale yellow solid which was taken
in methanol and stirred for 1 h in the presence of activated
charcoal and then filtered through Celite and evaporated to
give 2.00 g (18%) of a white solid. 1H NMR: (DMSO-d6, 300
MHz) δ 0.74 (t, J ) 7.4 Hz, 3H), 1.05-1.25 (m, 2H), 1.58-
1.68 (m, 1H), 1.91 (ddd, J ) 12.5, 5.7, 5.7 Hz, 1H), 2.13-2.30
(m, 2H), 2.87 (dd, J ) 9.7, 5.1 Hz, 1H), 3.22-3.33 (m, 1H),
3.48 (dd, J ) 9.9, 5.5 Hz, 1H), 3.86 (s, 3H), 3.93 (dd, J ) 8.1,
5.5 Hz, 1H), 7.13 (br d, J ) 8.8 Hz, 2H), 7.79 (br d, J ) 8.8 Hz,
2H). ESI MS: m/z (rel intensity) 358.2 ([M + H]+, 100), 380.1
([M + Na]+, 5). Analysis: C, H, N for C15H23N3O5S‚HCl.
1N-(4-Met h oxyb en zen esu lfon yl)-2(R)-N-h yd r oxyca r -
boxa m id o-4(S)-N-(3-p yr id ylm eth yl)-N-m eth a n esu lfon yl-
a m in op yr r olid in e (27). The title compound was prepared
according to the synthetic sequence described for compound
1
23. H NMR: (DMSO-d6, 300 MHz) δ 1.87 (dd, J ) 12.6, 7.5
Hz, 1H), 2.63 (ddd, J ) 12.2, 9.3 Hz, 1H), 2.86 (dd, J ) 9.3,
9.3 Hz, 1H), 3.05 (s, 3H), 3.60 (dd, J ) 8.5, 8.5 Hz, 1H), 3.90
(s, 3H), 3.98-4.13 (m, 2H), 4.28 (d, J ) 17.4 Hz, 2H), 4.58-
4.82 (m, 1H), 7.13 (ddd, J ) 9.0, 2.0, 2.0 Hz, 2H), 7.35 (dd, J
) 7.9, 4.8 Hz, 1H), 7.57 (ddd, J ) 7.9, 1.8, 1.8 Hz, 1H), 7.71
(ddd, J ) 9.0, 1.8, 1.8 Hz, 2H), 8.32 (d, J ) 1.8 Hz, 1H), 8.47
(dd, J ) 6.4, 1.7 Hz, 1H), 9.04 (br s, 1H), 10.73 (br s, 1H). ESI
MS: m/z (rel intensity) 484.9 ([M + H]+, 100), 506.9 ([M +
NH4]+, 10). Analysis: C, H, N for C19H24N4O7S2‚H2O.
1N-(4-Met h oxyb en zen esu lfon yl)-2(R)-N-h yd r oxyca r -
b oxa m id o-4(S)-N-m et h a n esu lfon yl-N-p r op yla m in op yr -
r olid in e (28). The starting ester 7 (614 mg, 1.41 mmol) was
converted to the title compound (448 mg, 73%) as described
1
for 23. H NMR: (DMSO-d6, 300 MHz) δ 0.68 (t, J ) 7.3 Hz,
1N-(4-Met h oxyb en zen esu lfon yl)-2(R)-N-h yd r oxyca r -
boxa m id o-4(S)-N-(2-p h en yleth yl)a m in op yr r olid in e (22).
The title compound was prepared as described for compound
21 and purified over flash silica with EtOAc:MeOH (4:1) to
give a white solid. 1H NMR: (DMSO-d6, 300 MHz) δ 1.58-
1.67 (m, 1H), 1.88-1.94 (m, 1H), 2.37-2.57 (m, 2H), 2.88 (dd,
J ) 9.7, 5.3 Hz, 1H), 3.48 (dd, J ) 10.0, 5.5 Hz, 1H), 3.69-
3.87 (m, 1H), 3.85 (s, 3H), 3.94 (dd, J ) 7.5, 5.7 Hz, 1H), 7.11-
7.10 (m, 5H), 7.82 (br d, J ) 9.0 Hz, 2H). ESI MS: m/z (rel
intensity) 420.4 ([M + H]+, 100). Analysis: C20H25N3O5S‚HCl‚
0.5H2O.
3H), 1.22-1.42 (m, 2H), 1.61-1.74 (m, 1H), 1.93 (dd, J ) 12.6,
7.0 Hz, 1H), 2.74 (ddd, J ) 14.8, 9.9, 5.7 Hz, 1H), 2.85-2.91
(m, 4H), 3.62 (dd, J ) 8.8, 8.8 Hz, 1H), 3.87 (s, 3H), 4.16 (d, J
) 9.2 Hz, 1H), 4.55-4.66 (m, 1H), 7.18 (br d, J ) 9.0 Hz, 2H),
7.84 (br d, J ) 9.0 Hz, 2H). ESI MS: m/z (rel intensity) 452.9
([M + NH4]+ 100), 435.8 ([M + H]+, 55). Analysis: C,H,N for
C
16H25N3O7S2.
1N-(4-Meth oxyben zen esu lfon yl)-2(R)-h yd r oxyca r box-
a m id o- 4(S )- 4- m e t h oxyp h e n ylsu lfon yla m in op yr r oli-
d in e (29). The desired material was prepared according to the
synthetic sequence described for compound 23. 1H NMR:
(DMSO-d6, 300 MHz) δ 1.54 (ddd, J ) 12.6, 9.3, 9,3 Hz, 1H),
1.68-1.76 (m, 1H), 2.78 (dd, J ) 8.6, 8.6 Hz, 1H), 3.36 (dd, J
) 9.2, 6.8 Hz, 1H), 3.73-3.83 (m, 1H), 3.85 (s, 3H), 3.87 (s,
3H), 3.92 (dd, J ) 9.0, 2.4 Hz, 1H), 7.12 (ddd, J ) 9.0, 2.6, 2.6
Hz, 2H), 7.16 (ddd, J ) 9.0, 2.6, 2.6 Hz, 2H), 7.69 (ddd, J )
9.0, 2.6, 2.6 Hz, 2H), 7.74 (ddd, J ) 9.0, 2.6, 2.6 Hz, 2H), 8.34
(s, 1H). ESI MS: m/z (rel intensity) 486.3 ([M + H]+, 100).
Analysis: C, H, N for C19H23N3O8S2‚0.7H2O.
1N-(4-Meth oxyben zen esu lfon yl)-2(R)-h yd r oxyca r box-
a m id o-4(S)-m eth a n esu lfon yla m in op yr r olid in e (23). The
starting ester 5 (354 mg, 0.903 mmol) was converted to the
title compound and chromatographed as described for com-
pound 3. It was then recrystallized from acetonitrile/water to
1
give 261 mg (74%) of pale yellow crystals. H NMR: (DMSO-
d6, 300 MHz) δ 1.67 (ddd, J ) 12.6, 6.3, 6.3 Hz, 1H), 2.04 (ddd,
J ) 12.6, 6.2, 2.2 Hz, 1H), 2.86 (dd, J ) 8.6, 8.6 Hz, 1H), 2.90

4958 J ournal of Medicinal Chemistry, 2000, Vol. 43, No. 26
Natchus et al.
1
1N-(4-n -Bu t oxyb en zen esu lfon yl)-2(R)-N-h yd r oxyca r -
boxa m id o -4(S)-(1N-m eth ylim id a zol-4-yl)su lfon yla m in o-
p yr r olid in e (30). The desired material was prepared accord-
a white soild. H NMR: (DMSO-d6, 300 MHz) δ 1.18 (dd, J )
6.6, 2.9 Hz, 3H), 1.75-2.0 (m, 2H), 2.88 (ddd, J ) 8.1, 8.1, 7.8
Hz, 1H), 3.55 (dd, J ) 8.4, 7.3 Hz, 1H), 3.76 (q, J ) 6.8 Hz,
1H), 3.85 (s, 3H), 4.03 (dd, J ) 8.4, 2.4 Hz, 1H), 4.14 (d, J )
12.3 Hz, 1H), 4.41-4.56 (m, 1H), 4.45 (d, J ) 11.9 Hz, 1H),
7.15 (br d, J ) 8.6 Hz, 2H), 7.26-7.39 (m, 5H), 7.80 (br d, J )
8.8 Hz, 2H), 7.84 (dd, J ) 7.9, 3.1 Hz, 1H), 8.98 (s, 1H), 10.66
(s, 1H). ESI MS: m/z (rel intensity) 478.3 ([M + H]+, 100),
500.2 ([M + NH4]+, 12). Analysis: C, H, N for C22H27N3O7S.
1
ing to the synthetic sequence described for compound 23. H
NMR: (DMSO-d6, 300 MHz) δ 0.97 (t, J ) 7.4 Hz, 3H), 1.39-
1.63 (m, 3H), 1.69-1.87 (m, 3H), 2.76 (dd, J ) 8.8, 8.8 Hz,
1H), 3.41 (dd, J ) 9.0, 6.8 Hz, 1H), 3.71 (s, 3H), 3.90-4.0 (m,
2H), 4.08 (t, J ) 12.8 Hz, 2H), 7.14 (br d, J ) 9.0 Hz, 2H),
4.11 (br d, J ) 9.0 Hz, 2H), 7.16 (s, 1H), 7.80 (d, J ) 0.9 Hz,
1H), 9.05 (br s, 1H), 10.7 (s, 1H). ESI MS: m/z (rel intensity)
502.2 ([M + H]+, 100). Analysis: C, H, N for C19H27N5O7S2‚
H2O.
1N-(4-Meth oxyben zen esu lfon yl)-2(R)-h yd r oxyca r box-
a m id o-4(S)-[(1,1′-b ip h en yl)-4-yl]ca r b on yla m in op yr r oli-
d in e (31). The desired material was prepared according to the
synthetic sequence described for compound 23. 1H NMR:
(DMSO-d6, 300 MHz) δ 2.08 (dd, J ) 6.5, 6.5 Hz, 2H), 3.21
(dd, J ) 9.9, 5.7 Hz, 1H), 3.64-3.70 (m, 4H), 4.13 (dd, J ) 7.0,
7.0 Hz, 1H), 4.44-4.55 (m, 1H), 7.01 (br d, J ) 9.0 Hz, 2H),
7.40-7.45 (m, 1H), 7.52 (dd, J ) 7.4, 7.4 Hz, 2H), 7.69-7.80
(m, 8H), 8.14 (d, J ) 5.7 Hz, 1H). ESI MS: m/z (rel intensity)
517.8 ([M + Na]+, 15), 513.0 ([M + NH4]+, 60), 496.0 ([M +
H]+, 100). Analysis: C, H, N for C25H25N3O6S‚2H2O.
1N-(4-Meth oxyben zen esu lfon yl)-2(R)-h yd r oxyca r box-
a m id o-4(S)-N-m eth ylca r boxa id oa m in op yr r olid in e (32).
The desired material was prepared according to the synthetic
sequence described for comound 23. 1H NMR: (DMSO-d6, 300
MHz) δ 1.63 (ddd, J ) 12.3, 8.8, 8.8 Hz, 1H), 1.93 (ddd, J )
12.6, 6.0, 4.0 Hz, 1H), 2.57 (d, J ) 4.8 Hz, 3H), 2.81 (dd, J )
9.0, 7.3 Hz, 1H), 2.52 (dd, J ) 9.2, 6.2 Hz, 1H), 3.87 (s, 3H),
3.98 (dd, J ) 8.8, 3.7 Hz, 1H), 4.15-4.26 (m, 1H), 5.54 (dd, J
) 4.8, 2.0 Hz, 1H), 5.88 (d, J ) 6.6 Hz, 1H), 7.15 (d, J ) 8.8
Hz, 2H), 7.78 (d, J ) 8.8 Hz, 2H), 9.01 (s, 1H), 10.8 (s, 1H).
ESI MS: m/z (rel intensity) 411.0 ([M + K]+, 30), 373.1 ([M +
H]+, 100), 316 (32). Analysis: C, H, N for C14H20N4O6S.
1N-(4-Met h oxyb en zen esu lfon yl)-2(R)-N-h yd r oxyca r -
boxa m id o-4(S)-N-(3-p yr id ylm eth yl)-N-isop r op ylca r bon -
yla m in op yr r olid in e (33). The title compound was prepared
according to the synthetic sequence described for compound
23. 1H NMR: (DMSO-d6, 300 MHz) δ 1.23 (d, J ) 6.6 Hz, 6H),
1.68 (dd, J ) 11.8, 7.2 Hz, 1H), 2.21-2.96 (m, 3H), 3.51 (dd, J
) 9.1, 7.8 Hz, 1H), 3.90 (s, 3H), 4.03-4.31 (m, 3H), 4.48-4.74
(m, 1H), 7.11 (ddd, J ) 9.0, 2.0, 2.0 Hz, 2H), 7.33 (dd, J ) 7.8,
4.8 Hz, 1H), 7.57 (m, 1H), 7.71 (ddd, J ) 9.0, 2.0, 2.0 Hz, 2H),
8.33 (d, J ) 1.7 Hz, 1H), 8.49 (dd, J ) 6.7, 1.9 Hz, 1H), 9.08
(br s, 1H), 10.25 (br s, 1H). ESI MS: m/z (rel intensity) 476.9
([M + H]+, 100). Analysis: C, H, N for C22H28N4O6S‚1.5H2O.
2(R)-Ben zyloxy-3-p h en ylp r op ion ic Acid (37a ). Sodium
hydride (2.9 g, 120 mmol) was washed 2× with hexane and
covered with 50 mL of DMF. The starting L-3-phenyllactic acid
(5 g, 30.1 mmol) was then added in portions and after fizzing
ceased, the mixture was heated to 55 °C for 1 h. The mixture
was then cooled to 0 °C and benzyl bromide (4.3 mL, 36.1
mmol) was added dropwise. The mixture was heated to 60 °C
for 3 h and then partitioned between hexane:EtOAc (1:1) and
1 N HCl. The organic layer was washed with brine, dried over
MgSO4, filtered and evaporated. The residue was chromato-
graphed over flash silica with hexane:EtOAc (9:1 to 0:1) to give
4.00 g (52%) of colorless oil. 1H NMR: (DMSO-d6, 300 MHz) δ
3.05 (dd, J ) 14.1, 8.4 Hz, 1H), 3.18 (dd, J ) 14.1, 4.2 Hz,
1H), 4.20 (dd, J ) 8.2, 1H), 4.43 (d, J ) 11.7 Hz, 1H), 4.65
(dd, J ) 11.5 Hz, 1H), 7.14-7.18 (m, 1H), 7.23-7.33 (m, 4H).
ESI MS: m/z (rel intensity) 274.3 ([M + NH4]+, 100).
1N-(4-Met h oxyb en zen esu lfon yl)-2(R)-ca r b om et h oxy-
4(S)-N-(1-oxy-2(R)-ben zyloxy-3-p h en ylp r op yl)a m in op yr -
r olid in e (37b). The starting amine 3 (800 mg, 2.55 mmol)
and the starting benzyl lactic acid 37a (784 mg, 3.06 mmol)
were taken in 5 mL of DMF in the presence of 1 mL of
N-methylmorpholine, 1-ethyl-3-(3-dimethylaminopropyl)car-
bodiimide (979 mg, 5.10 mmol) and hydroxybenzotriazole (1.03
mg, 7.65 mmol). The resulting mixture was stirred at room
temperature for 16 h and then partitioned between 1 N HCl
and EtOAc. The organic layer was then washed 1× with dilute
NaHCO3, 1× with brine, dried over MgSO4, filtered and
evaporated. The crude residue was then chromatographed with
hexane:EtOAc (8:1 to 1:1) to give 1.08 g (57%) of the title
compound. 1H NMR: (DMSO-d6, 300 MHz) δ 1.83-1.93 (m,
1H), 2.02-2.13 (m, 1H), 2.91 (dd, J ) 14.1, 6.6 Hz, 1H), 3.10-
3.25 (m, 2H), 3.51 (dd, J ) 10.8, 5.7 Hz, 1H), 3.72 (s, 3H), 3.86
(s, 3H), 4.07 (dd, J ) 6.6, 3.8 Hz, 1H), 4.23 (dd, J ) 8.1, 6.4
Hz, 1H), 4.38-4.53 (m, 3H), 6.48 (d, J ) 7.1 Hz, 1H), 6.96 (br
d, J ) 9.0 Hz, 2H), 7.19-7.40 (m, 10H), 7.81 (br d, J ) 9.0 Hz,
2H). ESI MS: m/z (rel intensity) 553.2 ([M + H]+, 100), 570.3
([M + NH4]+, 18).
1N-(4-Meth oxyp h en ylsu lfon yl)-2(R)-N-h yd r oxyca r box-
am ido-4(S)-N-(1-oxy-2(R)-ben zyloxy-3-ph en ylpr opyl)am i-
n op yr r olid in e (37). The starting methyl ester 37b (700 mg,
1.27 mmol) was taken in 2 mL of methanol, treated with NH2-
OK (2.5 mL, 1.25 M in methanol; solution prepared as
described in Fieser and Fieser, Vol 1, p 478) and stirred
overnight. The following morning, dry silica (3 mL) was added
to the mixture and the solvent removed under vaccuum. The
dry silica was poured on the top of a flash silica gel column
which was subsequently eluted with hexane:EtOAc (1:1 f 0:1)
to give 322 mg (46%) of a white foamy solid. 1H NMR: (DMSO-
d6, 300 MHz) δ 1.67 (dd, J ) 14.2, 7.5 Hz, 1H), 1.82-1.91 (m,
1H), 2.72 (dd, J ) 8.5, 8.5 Hz, 1H), 2.80 (d, J ) 6.0 Hz, 2H),
3.47 (dd, J ) 7.8, 7.8 Hz, 1H), 3.82 (s, 3H), 3.89 (dd, J ) 6.3,
6.3 Hz, 1H), 3.98 (d, J ) 7.3 Hz, 1H), 4.26 (d, J ) 12.3 Hz,
2H), 4.38-4.51 (m, 2H), 7.10-7.39 (m, 12 H), 7.77 (d, J ) 9.0
Hz, 2H), 7.87 (d, J ) 7.3 Hz, 1H), 9.00 (s, 1H), 10.7 (s, 1H).
ESI MS: m/z (rel intensity) 553.3 ([M + H]+, 100), 576.3 ([M
+ Na]+, 23). Analysis: C, H, N for C28H31N3O7S‚0.2H2O.
1N-(4-Met h oxyb en zen esu lfon yl)-2(R)-ca r b om et h oxy-
4(S)-N-(1-oxy-2(R)-ben zyloxyp r op yl)-N-p r op yla m in op yr -
r olid in e (38a ). The starting amine 4 (636 mg, 1.79 mmol) and
the starting benzyl lactic acid (390 mg, 2.15 mmol) were taken
in 5 mL of DMF in the presence of 1 mL of N-methylmorpho-
line, 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide (687 mg,
3.58 mmol) and hydroxybenzotriazole (762 mg, 5.37 mmol).
The resulting mixture was stirred at room temperature for
16 h and then partitioned between 1 N HCl and EtOAc. The
1N-(4-n -P r op oxyben zen esu lfon yl)-2(R)-N-h yd r oxyca r -
b oxa m id o-4(S )-(1-oxy-2-m e t h oxye t h yl)a m in op yr r oli-
d in e (34). The title compound was prepared as described for
1
compound 23 to give a white solid. H NMR: (DMSO-d6, 300
MHz) δ 1.00 (t, J ) 7.3 Hz, 3H), 1.72-1.77 (m, 4H), 2.94 (dd,
J ) 8.2 Hz, 1H), 3.22 (s, 3H), 3.52 (dd, J ) 8.8, 6.8 Hz, 1H),
3.65 (s, 2H), 3.98 (dd, J ) 8.1, 4.2 Hz, 1H), 4.04 (t, J ) 6.2 Hz,
2H), 4.38-4.51 (m, 1H), 7.14 (d, 8.8 Hz, 2H), 7.68 (d, J ) 7.1
Hz, 1H), 7.76 (d, J ) 8.5 Hz, 2H), 8.70 (br s, 1H), 10.35 (s,
1H). ESI MS: m/z (rel intensity) 416.1 ([M + H]+, 100), 433.1
([M + NH4]+, 10). Analysis: C, H, N for C17H25N3O7S.
1N-(4-n -Bu t oxyb en zen esu lfon yl)-2(R)-N-h yd r oxyca r -
b oxa m id o-4(S )-(1-oxy-2-m e t h oxye t h yl)a m in op yr r oli-
d in e (35). The title compound was prepared as described for
1
compound 23 to give a white solid. H NMR: (DMSO-d6, 300
MHz) δ 0.95 (t, J ) 7.3 Hz, 3H), 1.39-1.52 (m, 2H), 1.68-
1.79 (m, 2H), 1.83-1.96 (m, 2H), 2.94 (dd, J ) 9.2, 7.7 Hz,
1H), 3.22 (s, 3H), 3.52 (dd, J ) 9.2, 6.6 Hz, 1H), 3.63 (s, 2H),
3.98 (dd, J ) 7.7, 4.0 Hz, 1H), 4.07 (t, J ) 6.4 Hz, 2H), 4.37-
4.51 (m, 1H), 7.14 (br d, J ) 9.2 Hz, 2H), 7.68 (d, J ) 7.0 Hz,
1H), 7.76 (br d, J ) 8.8 Hz, 2H), 9.00 (br s, 1H), 10.68 (s, 1H).
ESI MS: m/z (rel intensity) 430.1 ([M + H]+, 100), 447.1 ([M
+ NH4]+, 20). Analysis: C, H, N for C18H27N3O7S.
1N-(4-Met h oxyb en zen esu lfon yl)-2(R)-N-h yd r oxyca r -
boxa m id o -4(S)-N -(1-oxy -2(R)-ben zyloxyp r op yl)a m in o-
p yr r olid in e (36). The desired material was prepared accord-
ing to the synthetic sequence described for comound 37 to give

New MMP Inhibitors from Functionalized Aminoprolines
J ournal of Medicinal Chemistry, 2000, Vol. 43, No. 26 4959
organic layer was then washed 1× with dilute NaHCO3, 1×
with brine, dried over MgSO4, filtered and evaporated. The
crude residue was then chromatographed with hexane:EtOAc
(8:1 to 1:1) to give 406 mg (44%) of the title compound. 1H NMR
indicates two distinct rotamers in a 2:3 ratio which complicates
spectral interpretation. 1H NMR: (DMSO-d6, 300 MHz) δ 0.75-
0.87 (m, 3H), 1.34-1.51 (m, 5H), 1.95-2.17 (m, 2H), 2.44-
2.56, 2.68-2.80 (m, 1H), 2.89-3.20 (m, 2H), 3.34-3.54 (m, 1H),
3.56-3.74 (m, 3H), 3.85, 3.87 (m, 3H), 4.14-4.61, 5.09-5.22
(m, 5H), 7.0 (br d, J - 9.0 Hz, 2H), 7.26-7.38 (m, 5H), 7.73-
7.85 (m, 2H). ESI MS: m/z (rel intensity) 519.3 ([M + H]+,
100), 536.3 ([M + NH3]+, 60).
1N-(4-Met h oxyb en zen esu lfon yl)-2(R)-ca r b om et h oxy-
4(S)-N-(1-oxo-2(R)-h yd r oxyp r op yl)-N-p r op yla m in op yr -
r olid in e (38b). The starting ether 38a (700 mg, 1.35 mmol)
was taken in 25 mL of methanol with catalytic 10% Pd-C and
H2SO4 and hydrogenated for 3 h at 54 psi in a Parr apparatus.
The material was then filtered through a pad of Celite,
evaporated to dryness and chromatographed over flash silica
to give 341 mg (59%) of clear gum. 1H NMR indicates two
distinct rotamers in a 2:3 ratio which complicates spectral
interpretation. 1H NMR: (DMSO-d6, 300 MHz) δ 0.93-0.96
(m, 3H), [1.28-1.29 (d, J ) 7.1 Hz, 2H), 1.50-1.64 (m, 2H),
2.09-2.22 (m, 1H), 2.48-2.62 (m, 1H), 2.95-3.28 (m, 3H),
3.45-3.69 (m, 2H), 3.73 (s, 3H), 3.90 (s, 3H), 4.29-4.66 (m,
3H), 7.02 (br d, J ) 8.8 Hz, 2H), 7.83 (br d, J ) 8.8 Hz, 2H).
ESI MS: m/z (rel intensity) 429.3 ([M + H]+, 100), 446.3 ([M
+ NH3]+, 12).
1N-(4-Met h oxyb en zen esu lfon yl)-2(R)-ca r b om et h oxy-
4(S)-N-(1-oxy-2(R)-h yd r oxyp r op yl)-N -p r op yla m in op yr -
r olid in e (38). The starting methyester 38b (331 mg, 0.771
mmol) was taken in 1 mL of methanol, treated with NH2OK
(1.23 mL, 1.25 M in methanol; solution prepared as described
in Fieser and Fieser, Vol 1, p 478) and stirred overnight. The
following morning, dry silica (3 mL) was added to the mixture
and the solvent removed under vaccuum. The dry silica was
poured on the top of a flash silica gel column which was
subsequently eluted with hexane:EtOAc (1:1 f 0:1) to give 300
mg (91%) of a white foamy solid. 1H NMR indicates two distinct
rotamers in a 1:1 ratio which complicates spectral interpreta-
tion. 1H NMR: (DMSO-d6, 300 MHz) δ 0.60-0.77 (m, 3H),
1.60-1.37 (m, 5H), 1.63-2.04 (m, 2H), 2.70-3.08 (m, 2H),
3.25-3.65 (m, 2H), 3.86 (s, 3H), 4.08-4.40 (m, 2H), 4.56-5.02
(m, 1H), 7.12-7.20 (m, 2H), 7.76-7.86 (m, 2H), 10.75 (s, 1H).
ESI MS: m/z (rel intensity) 519.3 ([M + H]+, 100), 536.3 ([M
+ NH4]+, 60). Analysis: C, N, N for C18H27N3O7S‚0.3H2O.
(m, 2H), 2.86 (dd, J ) 8.3, 8.3 Hz, 1H), 3.51 (dd, J ) 7.5, 7.5
Hz, 1H), 3.85 (dd, J ) 12.8, 6.2 Hz, 1H), 3.98 (dd, J ) 7.3, 3.3
Hz, 1H), 4.09 (t, J ) 6.1 Hz, 2H), 4.39-4.54 (m, 1H), 5.39 (br
s, 1H), 7.14 (br d, J ) 8.8 Hz, 2H), 7.71 (d, J ) 7.5 Hz, 1H),
7.77 (br d, J ) 8.6 Hz, 2H), 9.0 (br s, 1H), 10.74 (s, 1H). ESI
MS: m/z (rel intensity) 430.1 (M + H+, 100). Analysis: C, H,
N for C17H25N3O7S‚0.2H2O.
1N-(4-n -Bu t oxyb en zen esu lfon yl)-2(R)-N-h yd r oxyca r -
b oxa m id o-4(S)-N-(1-oxy-2(R)-h yd r oxy-3-m et h ylb u t yl)-
a m in op yr r olid in e (42). The title compound was described
according to the synthetic sequence described for compound
38. 1H NMR: (DMSO-d6, 300 MHz) δ 0.79 (d, J ) 6.8 Hz, 3H),
0.84 (d, J ) 6.9 Hz, 3H), 0.95 (t, J ) 7.3 Hz, 3H), 1.39-1.53
(m, 2H), 1.68-1.93 (m, 4H), 2.99 (dd, J ) 8.6, 8.6 Hz, 1H),
3.34 (s, 3H), 3.51 (dd, J ) 7.9 Hz, 1H), 3.58 (dd, J ) 4.7, 4.7
Hz, 1H), 3.98 (dd, J ) 8.6, 2.4 Hz, 1H), 4.08 (t, J ) 6.4 Hz,
2H), 4.44-4.58 (m, 1H), 5.27 (d, J ) 5.3 Hz, 1H), 7.13 (d, J )
9.0, 2H), 7.71 (d, J ) 7.7 Hz, 1H), 7.77 (d, J ) 8.8 Hz, 2H),
8.99 (s, 1H), 10.73 (s, 1H). ESI MS: m/z (rel intensity) 474.8
(M + NH4+, 20), 457.8 (M + H+, 100). Analysis: C, H, N for
C
20H31N3O7S‚0.3H2O.
1N-(4-n -Bu t oxyb en zen esu lfon yl)-2(R)-N-h yd r oxyca r -
b oxa m id o-4(S)-N-(1-oxy-2(R)-h yd r oxy-4-m et h ylp en t yl)-
a m in op yr r olid in e (43). The title compound was described
according to the synthetic sequence described for compound
1
38. H NMR: (DMSO-d6, 300 MHz) δ 0.84 (br d, J ) 6.4 Hz,
6H), 0.95 (t, J ) 8.2 Hz, 3H), 1.23-1.39 (m, 2H), 1.39-1.53
(m 2H), 1.64, 1.92 (m, 4H), 2.87 (dd, J ) 8.3, 8.3 Hz, 1H), 3.51
(dd, J ) 7.7 Hz, 1H), 3.71-3.79 (m, 1H), 3.97 (dd, J ) 5.8, 0.8
Hz, 1H), 4.08 (t, J ) 6.1 Hz, 2H), 4.39-4.54 (m 1H), 5.30 (d,
J ) 5.5 Hz, 1H), 7.14 (d, J ) 7.9 Hz, 2H), 7.73 (d, J ) 8.6 Hz,
1H), 7.76 (d, J ) 8.1 Hz, 2H), 9.00 (s, 1H), 10.73 (s, 1H). ESI
MS: m/z (rel intensity) 489.2 ([M + NH4]+, 10), 472.0 ([M +
H]+, 100). Analysis: C, H, N for C21H33N3O7S‚0.5H2O.
1N-(4-Met h oxyb en zen esu lfon yl)-2(R)-N-h yd r oxyca r -
boxam ido-4(S)-(piper idin -1N-yl)pyr r olidin e (44). The com-
pound was prepared according to the synthetic sequence
described for compound 48 to give a pale orange solid. 1H
NMR: (DMSO-d6, 300 MHz) δ 1.27-1.43 (m, 6H), 1.52 (ddd,
J ) 12.5, 9.9, 9.9 Hz, 1H), 1.89-1.96 (m, 1H), 2.17-2.31 (m,
4H), 2.74 (dd, J ) 8.7, 8.7 Hz, 1H), 2.94-3.08 (m, 1H), 3.63
(dd, J ) 8.7, 6.8 Hz, 1H), 3.87 (s, 3H), 3.98 (d, J ) 8.7 Hz,
1H), 7.16 (br d, J ) 9.0 Hz, 2H), 7.80 (br d, J ) 9.0 Hz, 2H).
ESI MS: m/z (rel intensity) 384 (M+ + H, 100), 406 (M+
+
Na, 82), 422 (M+ + K, 65). Analysis: C, H, N for C17H25N3O5S.
1N-(4-n -P r op oxyben zen esu lfon yl)-2(R)-N-h yd r oxyca r -
boxam ido-4(S)-(piper idin -1N-yl)pyr r olidin e (45). The com-
pound was prepared according to the synthetic sequence
described for compound 48 to give a pale orange solid. 1H
NMR: (DMSO-d6, 300 MHz) δ 0.99 (t, J ) 7.4 Hz, 3H), 1.26-
1.54 (m, 7H), 1.70-1.83 (m, 2H), 1.90 (dd, J ) 12.3, 6.0 Hz,
1H), 2.10-2.29 (m, 4H), 2.69 (dd, J ) 8.4 Hz, 1H), 2.88-3.01
(m, 1H), 3.59 (dd, J ) 7.5 Hz, 1H), 3.98 (d, J ) 9.0 Hz, 1H),
4.05 (t, J ) 6.4 Hz, 2H), 7.15 (br d, J ) 8.8 Hz, 2H), 7.78 (br
d, J ) 8.8 hz, 2H), 8.96 (s, 1H), 10.69 (s, 1H). ESI MS: m/z
(rel intensity) 412.1 (M+ + H, 100). Analysis: C, H, N for
1N-(4-Met h oxyb en zen esu lfon yl)-2(R)-N-h yd r oxyca r -
boxa m id o-4(S)-N-(1-oxo-2(R)-h yd r oxy-3-p h en ylp r op yl)-
N-p r op yla m in op yr r olid in e (39). The compound was pre-
pared as described for compound 38 to give a white foamy
solid. 1H NMR indicates two distinct rotamers in a 2:3 ratio
1
which complicates spectral interpretation. H NMR: (DMSO-
d6, 300 MHz) δ 0.58-0.66 (m, 3H), 0.78-1.90 (m, 4H), 2.21-
2.96 (m, 5H), 3.29-3.61 (m, 2H), 3.33 (s, 3H), 3.85 (s, 3H),
3.95-5.19 (m, 5H), 7.12-7.28 (m, 7H), 7.23-7.83 (m, 3H). ESI
MS: m/z (rel intensity) 506.3 ([M + H]+, 100), 526.3 ([M +
Na]+, 12). Analysis: C, H, N for C24H31N3O7S‚0.6H2O.
C
19H29N3O5S.
1N-(4-n -P r op oxyben zen esu lfon yl)-2(R)-N-h yd r oxyca r -
boxa m id o-4(S)-N-(1-oxy-2(R)-h yd r oxyp r op yl)a m in op yr -
r olid in e (40). The title compound was described according to
1N-(4-n -P en t ylb en zen esu lfon yl)-2(R)-N-h yd r oxyca r -
boxam ido-4(S)-(piper adin -1N-yl)pyr r olidin e (46). The com-
pound was prepared according to the synthetic sequence
described for compound 48. 1H NMR: (DMSO-d6, 300 MHz) δ
0.87 (t, J ) 6.6 Hz, 3H), 1.23-1.40 (m, 10H), 1.48 (ddd, J )
12.1, 9.5, 9.5 Hz, 1H), 1.55-1.67 (m, 2H), 1.89 (dd, J ) 11.7,
5.5 Hz, 1H), 2.08-2.25 (m, 4H), 2.69 (t, J ) 7.7 Hz, 2H), 2.74
(dd, J ) 8.6, 8.6 Hz, 1H), 3.60 (dd, J ) 7.3, 8.0 Hz, 1H), 3.98
(d, J ) 7.69 Hz, 1H), 7.47 (d, J ) 7.9 Hz, 2H), 7.76 (d, J ) 7.3
Hz, 2H), 8.97 (s, 1H), 10.70 (s, 1H). ESI MS: m/z (rel intensity)
424.1 ([M + H]+, 100). Analysis: C, H, N for C21H33N3O4S.
1
the synthetic sequence described for compound 38. H NMR:
(DMSO-d6, 300 MHz) δ 1.02 (t, J ) 7.1 Hz, 3H), 1.15 (d, J )
6.8 Hz, 3H), 1.69-1.92 (m, 4H), 2.88 (dd, J ) 8.4, 8.4 Hz, 1H),
3.53 (dd, J ) 7.7, 7.7 Hz, 1H), 3.80-4.17 (m, 4H), 4.40-4.44
(m, 1H), 5.19 (d, J ) 5.1 Hz, 1H), 7.15 (br d, J ) 8.8 Hz, 2H),
7.72 (d, J ) 7.0 Hz, 1H), 7.77 (br d, J ) 8.8 Hz, 2H), 9.0 (br s,
1H), 10.75 (br s, 1H). ESI MS: m/z (rel intensity) 415.6 (M +
H+, 100). Analysis: C, H, N for C17H25N3O7S‚0.2H2O.
1N-(4-n -Bu t oxyb en zen esu lfon yl)-2(R)-N-h yd r oxyca r -
boxa m id o-4(S)-N-(1-oxy-2(R)-h yd r oxyp r op yl)a m in op yr -
r olid in e (41). The title compound was described according to
the synthetic sequence described for compound 38. H NMR:
(DMSO-d6, 300 MHz) δ 0.95 (t, J ) 7.2 Hz, 3H), 1.13 (d, J )
6.6 Hz, 3H), 1.38-1.53 (m, 2H), 1.67-1.80 (m, 2H), 1.81-1.94
1N-(4-P h en yloxyben zen esu lfon yl)-2(R)-N-h yd r oxyca r -
boxam ido-4(S)-(piper adin -1N-yl)pyr r olidin e (47). The com-
pound was prepared according to the synthetic sequence
described for compound 48. 1H NMR: (DMSO-d6, 300 MHz) δ
1.25-1.43 (m, 6H), 1.58 (ddd, J ) 12.3, 9.9, 9.9 Hz, 1H), 1.92
(dd, J ) 12.1, 5.5 Hz, 1H), 2.12-2.29 (m, 4H), 2.78 (dd, J )
1

4960 J ournal of Medicinal Chemistry, 2000, Vol. 43, No. 26
Natchus et al.
8.3, 8.3 Hz, 1H), 2.90-3.02 (m, 1H), 3.61 (dd, J ) 8.1, 7.0 Hz,
1H), 3.98 (d, J ) 9.2 Hz, 1H), 7.12-7.21 (m, 2H), 7.29 (dd, J
) 7.3, 7.3 Hz, 1H), 7.49 (dd, 7.7, 7.7 Hz, 2H), 7.85 (dd, J )
8.8, 1.5 Hz, 2H), 8.97 (s, 1H), 10.70 (s, 1H). ESI MS: m/z (rel
intensity) 446.1 ([M + H]+, 100). Analysis: C, H, N for
synthetic sequence described for compound 48 and recrystal-
lized from EtOAC:methanol to white crystals. 1H NMR:
(DMSO-d6, 300 MHz) δ 1.62 (ddd, J ) 12.6, 9.3, 9.3 Hz, 1H),
1.91-1.99 (m, 1H), 2.72-2.78 (m, 4H), 2.83 (dd, J ) 8.4, 8.4
Hz, 1H), 2.94-2.99 (m, 4H), 3.29-3.39 (m, 1H), 3.62 (dd, J )
8.8, 7.2 Hz, 1H), 3.88 (s, 3H), 3.99 (dd, J ) 9.0, 2.4 Hz, 1H),
7.16 (br d, J ) 8.6 Hz, 2H), 7.82 (ddd, J ) 8.6 Hz, 2H), 9.00 (s,
1H), 10.71 (s, 1H). ESI MS: m/z (rel intensity) 434.0 ([M +
H]+, 100), 456.0 ([M + Na]+, 32). Analysis: C, H, N for
C
22H27N3O5S‚0.3H2O.
1N-(4-Meth oxyp h en ylsu lfon yl)-2(R)-N-h yd r oxyca r box-
a m id o-4(S)-(m or p h olin -1N-yl)p yr r olid in e (48). The start-
ing methyl ester 10 (310 mg, 0.86 mmol) was treated with
NH2OK (2 mL, 1.25 M in methalol) in 4 mL of methanol and
stirred overnight at room temperature. The material was then
condensed and partitioned between EtOAC and dilute NaH-
CO3. The organic layer was then dried over MgSO4, filtered
and evaporated. The residue was adsorbed onto silica and
eluted through a flash silica column with EtOAC:MeOH (1:0
to 4:1) to give 205 mg (66%) of material which was puffed to
C
16H23N3O7S2.
1N-(4-n -Bu t oxyb en zen esu lfon yl)-2(R)-N-h yd r oxyca r -
b oxa m id o-4(S)-(4,4-d ioxyt h iom or p h olin -1N-yl)p yr r oli-
d in e (54). The compound was prepared according to the
synthetic sequence described for compound 48 to give material
which was puffed to a white solid under vacuum and not
1
recrystallized. H NMR: (DMSO-d6, 300 MHz) δ 0.96 (t, J )
1
a white solid under vacuum and not recrystallized. H NMR:
7.3 Hz, 3H), 1.39-1.53 (m, 2H), 1.55-1.67 (m1H), 1.69-1.79
(m, 2H), 1.89-2.00 (m, 1H), 2.72-2.87 (m, 5H), 2.93-3.03 (m,
4H), 3.30-3.40 (m, 1H), 3.61 (dd, J ) 8.6, 6.2 Hz, 1H), 3.99
(br d, J ) 7.0 Hz), 4.09 (t, J ) 6.4 Hz, 2), 7.15 (br d, J ) 8.6
Hz, 2H), 7.79 (br d, J ) 8.6 Hz, 2H). ESI MS: m/z (rel
intensity) 476.1 ([M + H]+, 100), 498.1 ([M + Na]+, 22).
Analysis: C, H, N for C19H29N3O6S2.
(DMSO-d6, 300 MHz) δ 1.53 (ddd, J ) 12.5, 8.8, 8.8 Hz, 1H),
1.88-1.96 (m, 1H), 2.17-2.31 (m, 4H), 2.78 (dd, J ) 8.4, 8.4
Hz, 1H), 2.90-3.00 (m, 1H), 3.41-3.46 (m, 4H), 3.59 (dd, J )
9.0, 6.4 Hz, 1H), 3.88 (s, 3H), 3.98 (d, J ) 6.78 Hz, 1H), 7.16
(br d, J ) 9.0 Hz, 2H), 7.81 (br d, J ) 9.0 Hz, 2H). ESI MS:
m/z (rel intensity) 408.1 ([M + NH4]+, 7), 386.1 ([M + H]+,
100). Analysis: C, H, N for C16H23N3O6S‚0.6H2O.
1N-(4-n -P r op oxyben zen esu lfon yl)-2(R)-N-h yd r oxyca r -
boxa m id o-4(S)-(1-p yr r olid in -1N-yl)p yr r olid in e (55). The
compound was prepared according to the synthetic sequence
described for compound 57 to give a white solid. 1H NMR:
(DMSO-d6, 300 MHz) δ 0.99 (t, J ) 7.3 Hz, 3H), 1.46-1.56
(m, 4H), 1.63 (ddd, J ) 12.1, 8.4, 8.4 hz, 1H), 1.69-1.82 (m,
2H), 1.88 (ddd, J ) 13.1, 4.0, 4.0 Hz, 1H), 2.25-2.34 (m, 4H),
2.76-2.70 (m, 2H), 3.46-3.55 (m, 1H), 3.96 (dd, J ) 8.4, 4.0
Hz, 1H), 4.03 (t, J ) 6.4 Hz, 2H), 7.12 (d, J ) 8.8 Hz, 2H),
7.78 (d, J ) 8.8 Hz, 2H), 8.97 (br s, 1H), 10.69 (s, 1H). ESI
MS: m/z (rel intensity) 398.1 ([M + H]+, 100). Analysis: C,
H, N for C18H27N3O5S‚0.2H2O.
1N-(4-Flu or oph en oxyben zen esu lfon yl)-2(R)-N-h ydr oxy-
ca r boxa m id o-4(S)-(p yr r olid in -1N-yl)p yr r olid in e (56). The
compound was prepared according to the synthetic sequence
described for compound 57. 1H NMR: (DMSO-d6, 300 MHz) δ
1.48-1.58 (m, 4H), 1.73 (ddd, J ) 12.5, 7.9, 7.9 Hz, 1H), 1.91
(ddd, J ) 12.5, 4.9 Hz, 1H), 2.18-2.36 (m, 4H), 2.80-2.88 (m,
1H), 2.95 (dd, J ) 9.3, 6.4 Hz, 1H), 3.53 (dd, J ) 9.2, 5.3 Hz,
1H), 3.98 (dd, J ) 8.2, 4.4 Hz, 1H), 7.13 (d, J ) 8.4 Hz, 2H),
7.23 (dd, J ) 8.2, 4.8 Hz, 2H), 7.33 (dd, J ) 8.2, 8.2 Hz, 2H),
7.85 (d, J ) 7.84 Hz, 2H), 8.97 (s, 1H), 10.71 (s, 1H). ESI MS:
m/z (rel intensity) 449.9 ([M + H]+, 100). Analysis: C, H, N
for C21H24FN3O5S‚0.4H2O.
1N-(4-n -P r op oxyben zen esu lfon yl)-2(R)-N-h yd r oxyca r -
boxa m id o-4(S)-(m or p h olin -1N-yl)p yr r olid in e (49). The
compound was prepared according to the synthetic sequence
1
described for compound 48. H NMR: (DMSO-d6, 300 MHz) δ
1.01 (t, J ) 7.3 Hz, 3H), 1.55 (ddd, J ) 12.6, 6.1, 6.1 Hz,
1H),1.72-1.84 (m, 2H),), 1.93 (ddd, J ) 12.5, 6.6, 2.4 Hz, 1H),
2.17-2.32 (m 4H), 2.78 (dd, J ) 8.5, 8.5, Hz, 1H), 2.89-3.00
(m, 1H), 3.41-3.48 (m, 4H), 3.60 (dd, J ) 8.8, 6.2 Hz, 1H),
3.97 (dd, J ) 8.8, 2.2 Hz, 1H), 4.06 (t, J ) 6.6 Hz, 2H), 7.16 (d,
J ) 9.0 Hz, 2h), 7.79 (d, J ) 9.0 Hz, 2H), 8.98 (d, J ) 1.5 Hz,
1H), 10.71 (d J ) 1.5 Hz, 1H). ESI MS: m/z (rel intensity)
414.1 ([M + H]+, 100). Analysis: C,H,N for C18H27N3O6S.
1N-(4-n -Bu t oxyb en zen esu lfon yl)-2(R)-N-h yd r oxyca r -
boxa m id o-4(S)-(m or p h olin -1N-yl)p yr r olid in e (50). The
compound was prepared according to the synthetic sequence
described for compound 48 to give material which was puffed
to a white solid under vacuum and not recrystallized. 1H
NMR: (DMSO-d6, 300 MHz) δ 0.96 (t, J ) 7.4 Hz, 3H), 1.39-
1.59 (m, 3H), 1.69-1.77 (m, 2H), 1.87-1.95 (m, 1H), 2.16-
2.31 (m, 4H), 2.77 (dd, J ) 8.4, 8.4 Hz, 1H), 2.89-3.00 (m,
1H), 3.40-3.46 (m, 4H), 3.58 (dd, J ) 8.6, 6.0 Hz, 1H), 3.97
(dd, J ) 8.5, 2.5 Hz, 1H), 4.08 (t, J ) 6.4 Hz, 2H), 7.14 (br d,
J ) 8.8 Hz, 2H), 7.78 (br d, J ) 8.9 Hz, 2H). ESI MS: m/z (rel
intensity) 428.08 ([M + H]+, 100), 450.07 ([M + Na]+, 8), 465.99
([M + K]+, 15). Analysis: C, H, N for C19H29N3O6S‚0.1H2O.
1N-(4-n -P en t ylb en zen esu lfon yl)-2(R)-N-h yd r oxyca r -
boxa m id o-4(S)-(m or p h olin -1N-yl)p yr r olid in e (51). The
compound was prepared according to the synthetic sequence
described for compound 48. 1H NMR: (DMSO-d6, 300 MHz) δ
0.87 (t, J ) 6.3 Hz, 3H), 1.22-1.37 (m, 4H), 1.49-1.67 (m, 3H),
1.91 (br d, J ) 12.5 Hz, 1H), 2.14-2.29 (m, 4H), 2.69 (dd, J )
7.5, 7.5 Hz, 1H), 2.80-2.99 (m, 2H), 3.36-3.44 (m, 4H), 3.60
(dd, J ) 8.2, 6.2 Hz, 1H), 3.98 (d, J ) 7.3 Hz, 1H), 7.46 (d, J
) 7.2 Hz, 2H), 7.77 (d, J ) 7.0 Hz, 2H), 8.98 (s, 1H), 10.71 (s,
1H). ESI MS: m/z (rel intensity) 426.1 ([M + H]+, 100).
Analysis: C, H, N for C20H31N3O5S.
1N-(4-P h en yloxyben zen esu lfon yl)-2(R)-N-h yd r oxyca r -
boxa m id o-4(S)-(p yr r olid in -1N-yl)p yr r olid in e (57). The
starting methyl ester 16 (752 mg, 1.75 mmol) was converted
to the title hydroxamic acid as described for compound 48 and
purified by recrystallizing from EtOAc:MeOH (∼10:1) to give
320 mg (43%) of white powder. 1H NMR: (DMSO-d6, 300 MHz)
δ 1.50-1.58 (m, 4H), 1.69-1.80 (m, 1H), 1.87-1.75 (m, 1H),
2.19-2.37 (m, 4H), 2.80-2.90 (m, 1H), 2.76 (dd, J ) 9.3, 6.2
Hz, 1H), 3.53 (dd, J ) 9.5, 6.2 Hz, 1H), 3.88 (dd, J ) 7.7, 4.8
Hz, 1H), 7.10-7.19 (m, 4H), 7.29 (ddd, J ) 7.3, 7.3, 1.3 Hz,
1H), 7.50 (dd, J ) 7.1, 7.1 Hz, 2H), 7.86 (dd, J ) 8.6, 1.7 Hz,
2H), 8.99 (br s, 1H), 10.71 (br s, 1H). ESI MS: m/z (rel
intensity) 432.2 ([M + H]+, 100). Analysis: C, H, N for
C
21H25N3O5S‚0.2H2O.
1N-(4-P h en yloxyben zen esu lfon yl)-2(R)-N-h yd r oxyca r -
boxa m id o-4(S)-(m or p h olin -1N-yl)p yr r olid in e (52). The
compound was prepared according to the synthetic sequence
described for compound 48. 1H NMR: (DMSO-d6, 300 MHz) δ
1.57-1.72 (m, 1H), 1.87-1.98 (m, 1H), 2.16-2.34 (m, 4H),
2.82-3.00 (m, 2H), 3.38-3.50 (m, 4H), 3.56-3.64 (m, 1H),
3.94-4.01 (m, 1H), 7.11-7.20 (m, 4H), 7.28 (ddd, J ) 6.9, 6.9,
0.9 Hz, 1H), 7.49 (ddd, J ) 7.6, 7.6, 1.1 Hz, 1H), 7.86 (dd, J )
8.6 Hz, 2H), 8.99 (s, 1H), 10.71 (s, 1H). ESI MS: m/z (rel
intensity) 448.0 ([M + H]+, 100). Analysis: C, H, N for
1N-(4-Met h oxyb en zen esu lfon yl)-2(R)-N-h yd r oxyca r -
b oxa m id o-4(S)-(γ-su lt a m -1N-yl)p yr r olid in e (58). Com-
pound 24 was prepared as described for compound 23 to give
a white solid. 1H NMR: (DMSO-d6, 300 MHz) δ 1.83-2.17 (m,
4H), 2.91-3.03 (m, 2H), 3.06 (dd, J ) 9.5, 8.1 Hz, 1H), 3.12-
3.22 (m, 2H), 2.79-3.89 (m, 2H), 3.59 (dd, J ) 9.7, 6.8 Hz,
1H), 3.87 (s, 3H), 3.92-4.03 (m, 1H), 4.05 (dd, J ) 8.4, 2.7 Hz,
1H), 7.17 (br d, J ) 9.0 Hz, 2H), 7.82 (br d, J ) 9.0 Hz, 2H),
9.02 (s, 1H), 10.81 (s, 1H). ESI MS: m/z (rel intensity) 420.0
([M + H]+, 100), 437 ([M + NH4]+, 20). Analysis: C, H, N for
C
19H29N3O6S‚0.2H2O.
C
15H21N3O7S2.
1N-(4-Met h oxyb en zen esu lfon yl)-(2R )-N-h yd r oxyca r -
b oxa m id o-4(S)-(4,4-d ioxyt h iom or p h olin -1N-yl)p yr r oli-
d in e (53). The compound was prepared according to the
1N-(4-n -P r op oxyben zen esu lfon yl)-2(R)-N-h yd r oxyca r -
b oxa m id o-4(S)-(γ-su lt a m -1N-yl)p yr r olid in e (59). Com-

New MMP Inhibitors from Functionalized Aminoprolines
J ournal of Medicinal Chemistry, 2000, Vol. 43, No. 26 4961
pound 24 was prepared as described for compound 23 to give
a white solid. H NMR: (DMSO-d6, 300 MHz) δ 1.00 (t, J )
synthetic sequence described for compound 61 to give a white
powder. 1H NMR: (DMSO-d6, 300 MHz) δ 1.01 (d, J ) 6.8 Hz,
6H), 1.84 (dd, J ) 11.2, 7.7 Hz, 1H), 2.00-2.13 (m, 1H), 2.44
(ddd, J ) 13.0, 9.9, 9.9 Hz, 1H), 2.78 (s, 3H), 3.48 (d, J ) 8.2
Hz, 2H), 3.80 (s, 2H), 3.86 (d, J ) 6.4 Hz, 2H), 4.08 (d, J ) 8.6
Hz, 1H), 4.70-4.83 (m, 1H), 7.15 (br d, J ) 8.8 Hz, 2H), 7.76
(br d, J ) 8.8 Hz, 2H), 9.12 (br s, 1H), 10.78 (br s, 1H). ESI
1
7.4 Hz, 3H), 1.70-1.83 (m, 2H), 1.84-2.15 (m, 4H), 2.91-3.24
(m, 6H), 3.60 (dd, J ) 9.7, 6.8 Hz, 1H), 3.92-4.09 (m, 3H),
7.16 (d, J ) 9.0 Hz, 2H), 7.80 (d, J ) 9.0 Hz, 2H), 9.04 (s, 1H),
10.82 (s, 1H). ESI MS: m/z (rel intensity) 448.0 ([M + H]+,
90), 465.1 ([M + NH4]+, 100). Analysis: C, H, N for C17H25
N3O7S2.
-
MS: m/z (rel intensity) 455.1 ([M + H]+, 100), 472.1 ([M+
NH4]+, 15). Analysis: C, H, N for C19H26N4O7S‚0.7H2O.
+
1N-(4-n -Bu t oxyb en zen esu lfon yl)-2(R)-N-h yd r oxyca r -
boxa m id o-4(S)-(γ-su lta m -1N-yl)p yr r olid in e (60). The com-
pound was prepared according to the synthetic sequence
described for compound 59. 1H NMR: (DMSO-d6, 300 MHz) δ
0.95 (t, J ) 7.4 Hz, 3H), 1.39-1.52 (m, 2H), 1.68-1.79 (m, 2H),
1.83-2.18 (m, 4H), 2.91-3.23 (m, 6H), 3.59 (dd, J ) 9.7, 6.8
Hz, 1H), 3.92-4.12 (m, 3H), 7.15 (br d, J ) 9.0 Hz, 2H), 7.80
(br d, J ) 9.0 Hz, 2H), 9.04 (s, 1H), 10.81 (d, J ) 1.1 Hz, 1H).
ESI MS: m/z (rel intensity) 462.0 ([M + H]+, 90), 479.1 ([M +
NH4]+, 100). Analysis: C, H, N for C18H27N3O7S2.
1N-(4-(2-Meth oxyeth oxy)ben zen esu lfon yl)-2(R)-N-h y-
d r oxyca r b oxa m id o-4(S)-1N-(3N-m et h ylh yd a n t oin -1N-
yl)p yr r olid in e (66). The compound was prepared according
to the synthetic sequence described for compound 61. The
product was recrystallized from cold methanol to give a white
1
powder. H NMR: (DMSO-d6,300 mHz) δ 1.80-1.87 (m, 1H),
2.37-2.45 (m, 1H), 2.77 (s, 3H), 3.32 (s, 3H), 3.48 (d, J ) 8.3,
2H), 3.68-3.71 (m, 2H), 3.79 (s, 2H), 4.11-4.22 (m, 2H), 4.72-
4.78 (m, 1H), 7.14-7.17 (m, 2H), 7.74-7.77 (m, 2H). ESI MS:
m/z (rel intensity) 457.08 ([M + H]+, 100), 474.09 ([M + NH4]+,
60). Analysis: C, H, N for C18H24N4O8S‚1.2H2O.
1N-(4-Met h oxyb en zen esu lfon yl)-2(R)-N-h yd r oxyca r -
b oxa m id o-4(S)-1N-(3N-m et h ylh yd a n t oin -1N-yl)p yr r oli-
d in e (61). The starting methyl ester 9 (500 mg, 1.22 mmol)
was taken in 7 mL of methanol/tetrahydrofuran (1:1), and
treated with NH2OK (2.5 mL, 1.25 M in methanol) and stirred
overnight. The following morning, dry silica (1.5 mL) was
added to the mixture and the solvent removed under vacuum.
The dry silica was poured on top of a flash silica column which
was subsequently eluted with ethyl acetate followed with ethyl
acetate (9:1) to give a clear glass which was puffed to a foamy
solid by slight heating under vacuum. The product was
recrystallized from cold methanol to give 428 mg (85%) of white
1N-(4-P h en oxyb en zen esu lfon yl)-2(R)-N-h yd r oxyca r -
b oxa m id o-4(S)-1N-(3N-m et h ylh yd a n t oin -1N-yl)p yr r oli-
d in e (67). The compound was prepared according to the
synthetic sequence described for compound 61. The product
was recrystallized from cold methanol to give a white powder.
1
H NMR: (DMSO-d6, 300 mHz) δ 1.87 (dd, J ) 11.5, 6.6 Hz,
1H), 2.36-2.47 (m, 1 H), 2.8 (s, 3H), 3.48 (d, J ) 8.9, 2H), 3.83
(s, 2H), 4.18 (br s, 1H), 4.79-4.85 (m, 1H), 7.16-7.19 (m, 4H),
7.26 (t, J ) 7.5 Hz, 1H), 7.45-7.50 (m, 2H), 7.8 (d, J ) 8.2,
2H). ESI MS: m/z (rel intensity) 475.09 ([M + H]+, 100), 497.07
([M + NH4]+, 60). Analysis: C, H, N for C21H22N4O7S‚1.9H2O.
1
powder. H NMR: (DMSO-d6, 300 mHz) δ 1.82-1.88 (m, 1H),
2.39-2.46 (m, 1H), 2.78 (s, 3H), 3.49 (d, J ) 3.49 Hz, 2H),
3.81 (s, 2H), 3.85 (s, 3H), 4.10 (d, J ) 8.6 Hz, 1H), 4.74-4.80
(m, 1H), 7.16 (d, J ) 8.79 Hz, 2H), 7.78 (d, J ) 8.8 Hz, 2H),
9.05 (s, 1H). ESI MS: m/z (rel intensity) 413.0 ([M + H]+, 100),
430.0 ([M + NH4]+, 55). Analysis: C, H, N for C16H20N4O7S‚
1.1H2O.
1N-(4-P yr id in -4-yloxyben zen esu lfon yl)-2(R)-N-h yd r ox-
yca r boxa m id o-4(S)-(3N-m eth ylh yd a n toin -1N-yl)p yr r oli-
d in e (68). The compound was prepared according to the
synthetic sequence described for compound 61 to give a white
solid. 1H NMR: (DMSO-d6, 300 mHz) δ 1.88 (dd, J ) 12.4, 7.1
Hz, 1H), 2.41 (ddd, J ) 10.9, 10.9, 10.6 Hz, 1H), 2.80 (s, 3H),
3.46 (dd, J ) 8.9, 8.9 Hz, 1H), 5.56 (dd, J ) 8.4, 8.4 Hz, 1H),
3.84 (s, 2H), 4.23 (d, J ) 8.4 Hz, 1H0, 4.75-4.89 (m, 1H), 7.11
(dd, J ) 4.8, 1.7 Hz, 2H), 7.42 (d, J ) 8.8 Hz, 2H), 7.93 (d, J
) 8.8 Hz, 2H), 8.53 (dd, J ) 4.8, 1.5 Hz, 2H), 9.25 (br s, 1H).
ESI MS: m/z (rel intensity) 476.0 ([M + H]+, 100). Analysis:
C, H, N for C20H21N5O7S‚1.2H2O.
1N-(4-n -Bu t oxyb en zen esu lfon yl)-2(R)-N-h yd r oxyca r -
b oxa m id o-4(S)-1N-(4S-m et h ylh yd a n t oin -1N-yl)p yr r oli-
d in e (69). The compound was prepared according to the
synthetic sequence described for compound 61. The product
was recrystallized from cold methanol to give a white powder.
1H NMR: (DMSO-d6, 300 MHz) δ 0.95 (t, J ) 7.3 Hz, 3H),
1.18 (d, J ) 6.9 Hz, 3H), 1.46 (dd, J ) 15.2, 7.5 Hz, 2H), 1.70-
1.88 (m, 3H), 2.41 (m, 1H), 3.33 (s, 3H), 3.48 (d, J ) 8.43 Hz,
2H), 3.91-3.96 (m, 1H), 4.07-4.10 (m, 3H), 7.14 (d, J ) 9.0
Hz, 2H), 7.76 (d, J ) 8.9 Hz, 2H), 8.22 (s,1H). ESI MS: m/z
(rel intensity) 455.0 (M+ + H, 100), 472.0 (M+ + NH3, 30).
HRMS: calcd for C19H27N4O7S, 455.1601; found, 455.1600.
1N-(4-n -Bu t oxyb en zen esu lfon yl)-2(R)-N-h yd r oxyca r -
boxa m id o-4(S)-1N-(4-d im eth ylh yd a n toin -1N-yl)p yr r oli-
d in e (70). The compound was prepared according to the
synthetic sequence described for compound 61. The product
was recrystallized from cold methanol to give a white powder.
1H NMR: (DMSO-d6, 300 MHz) δ 0.95 (t, J ) 7.36 Hz, 3H),
1.25 (s, 6H), 1.46 (dd, J ) 15.2, 7.5 Hz, 2H), 1.70-1.76 (m,
2H), 1.84-1.90 (m, 1H), 2.51-2.52 (m, 1H), 3.34 (s, 3H), 3.46-
3.50 (m, 2H), 4.08 (t, J ) 6.4, 3H), 7.14 (d, J ) 9.0 Hz, 2H),
7.76 (d, J ) 9.0 Hz, 2H), 8.31 (s,1H). ESI MS: m/z (rel
intensity) 469.0 ([M + H]+, 100), 486.0 ([M + NH4]+, 10).
Analysis: C, H, N for C20H28N4O7S‚0.7H2O.
1N-(4-n -P r op oxyp h en ylsu lfon yl)-2(R)-N-h yd r oxyca r -
b oxa m id o-4(S )-1N -(3N -a llylh yd a n t oin -1N -yl)p yr r oli-
d in e (71). The compound was prepared according to the
synthetic sequence described for compound 61 to give a white
powder. 1H NMR: (DMSO-d6, 300 MHz) δ 1.01 (t, J ) 7.3 Hz,
3H), 1.74-1.92 (m, 3H), 2.45 (ddd, J ) 11.0, 10.7, 10.7 Hz,
1H), 3.50 (d, J ) 8.6, 2H), 3.78 (s, 2H), 3.82-3.88 (m, 2H),
1N-(4-Eth oxyben zen esu lfon y)l-2(R)-N-h yd r oxyca r box-
a m id o-4(S)-1N-(3N-m eth ylh yd a n toin -1N-yl)p yr r olid in e
(62). The title compound was prepared as described for
1
compound 61. H NMR: (DMSO-d6, 300 MHz) δ 1.37 (t, J )
7.0 Hz, 3H), 1.84 (dd, J ) 11.0, 7.3 Hz, 1H), 2.43 (ddd, J )
11.4, 11.4, 10.8 Hz, 1H), 2.78 (s, 3H), 3.47 (s, 1H), 3.49 (s, 1H),
3.80 (s, 2H), 4.08 (d, J ) 7.5 Hz, 1H), 4.14 (q, J ) 7.0 Hz, 2H),
4.70-4.82 (m, 1H), 7.14 (d, J ) 9.0 Hz, 2H), 7.76 (d, J ) 9.0
Hz, 2H), 9.03 (br s, 1H), 10.80 (br s, 1H). ESI MS: m/z (rel
intensity) 427.0 ([M + H]+, 100), 444.0 ([M + NH4]+, 12), 449.0
([M + Na]+, 8). Analysis: C, H, N for C17H22N4O7S‚0.2H2O.
1N-(4-n -P r op oxyben zen esu lfon yl)-2(R)-N-h yd r oxyca r -
b oxa m id o-4(S)-1N-(3N-m et h ylh yd a n t oin -1N-yl)p yr r oli-
d in e (63). The title compound was prepared as described for
compound 61. The product was recrystallized from cold
methanol to give white powder. 1H NMR: (DMSO-d6, 300
MHz) δ 1.01 (t, J ) 7.4 Hz, 1H), 1.73-1.89 (m, 3H), 2.43 (dd,
J ) 10.2, 10.2 Hz, 1H), 2.78 (s, 3H), 3.47 (s, 1H), 3.50 (s, 1H),
3.80 (s, 2H), 4.01-4.12 (m, 3H), 4.69-4.82 (m, 1H), 7.14 (d, J
) 8.8 Hz, 2H), 7.75 (d, J ) 8.8 Hz, 2H), 9.05 (br s, 1H), 10.80
(br s, 1H). ESI MS: m/z (rel intensity) 458.1 ([M + NH4]+, 15),
441.0 ([M + H]+, 100). Analysis: C, H, N for C18H24N4O7S‚
0.5H2O.
1N-(4-n -Bu t oxyb en zen esu lfon yl)-2(R)-N-h yd r oxyca r -
b oxa m id o-4(S)-1N-(3N-m et h ylh yd a n t oin -1N-yl)p yr r oli-
d in e (64). The title compound was prepared as described for
compound 61. The product was recrystallized from cold
methanol to give white powder. 1H NMR: (DMSO-d6, 300
MHz) δ 0.95 (dd, J ) 8.2, 1.10 Hz, 3H), 1.46 (dd, J ) 14.8, 7.3
Hz, 2H), 1.70-1.87 (m, 2H), 2.39-2.48 (m, 1H), 2.78 (s, 3H),
3.48 (d, J ) 8.4 Hz, 2H), 3.80 (s, 2H), 4.00-4.73 (m, 3H), 4.74-
4.79(m, 1H), 7.14 (d, J ) 8.8 Hz, 2H), 7.76 (d, J ) 8.6 Hz, 2H).
ESI MS: m/z (rel intensity) 455.0 ([M + H]+, 100), 472.0 ([M
+ NH4]+, 50). Analysis: C, H, N for C19H26N4O7S‚0.4H2O.
1N-(4-Isobu toxyben zen esu lfon yl)-2(R)-N-h yd r oxyca r -
b oxa m id o-4(S)-1N-(3N-m et h ylh yd a n t oin -1N-yl)p yr r oli-
d in e (65). The compound was prepared according to the

4962 J ournal of Medicinal Chemistry, 2000, Vol. 43, No. 26
Natchus et al.
4.01-4.12 (m, 3H), 4.72-4.85 (m, 1H), 5.14-5.27 (m, 2H),
5.69-5.83 (m, 1H), 7.15 (dd, J ) 9.0, 1.8 Hz, 2H), 7.76 (dd, J
) 8.8, 1.8 Hz, 2H), 9.04 (br s, 1H), 10.78 (s, 1H). ESI MS: m/z
(rel intensity) 467.0 ([M + H]+, 100), 484.0 ([M + NH4]+, 10).
Analysis: C, H, N for C20H26N4O7S‚0.5H2O.
1N-(4-n -Bu t oxyb en zen esu lfon yl)-2(R)-N-h yd r oxyca r -
b oxa m id o-4(S )-1N -(3N -a llylh yd a n t oin -1N -yl)p yr r oli-
d in e (72). The compound was prepared according to the
synthetic sequence described for compound 61. The product
was recrystallized from cold methanol to give a white powder.
MMP-3 in the assay was 0.038 nM. The low concentration of
MMP-3 in the final MMP-9 dilution did not contribute to the
rate of substrate cleavage as assessed by control experiments
with 0.038 nM MMP-3. MMP-1, -3, -7, -8, and -13 were used
at final concentrations of 8, 16, 2, 4 and 0.5 nM, respectively.
The MMP assays was performed using the fluorogenic sub-
strate Mca-Pro-Leu-Gly-Leu-Dpa-Ala-Arg-NH2 at a concen-
tration of 4 µM at 25 °C. The assay buffer was 50 mM Tris,
pH 7.5, 0.2 M NaCl, 5 mM CaCl2 and 0.02% Brij-35. The
increase in fluorescence due to cleavage of the substrate (Gly-
Leu bond) was monitored kinetically for 30 min with a BMG
Fluostar fluorescence plate reader (λex 328 nm, λem 393 nm).
Each 96-well microtiter plate contained 100 µL of substrate
and 50 µL of enzyme in each well. 50 µL of MMP inhibitor
was added to each well (except for positive control) to give a
final volume of 200 µL/well. MMP inhibitors were tested at 8
different concentrations and an IC50 was calculated using the
formula: Vi/Vo ) 1/(1 + [I]/IC50), where Vi is the initial velocity
of substrate cleavage in the presence of inhibitor at concentra-
tion [I] and Vo is the initial velocity in the absence of inhib-
itor.
X-r a y Cr ysta llogr a p h y w ith Tr u n ca ted Str om elysin .
A truncated form of stromelysin containing the catalytic
domain with residues 83-255 was crystallized as described.32
1 mM compound 24 was added to the crystal hanging drop
for 4.5 h. Data were then collected at the IMCA beamline at
the Advanced Photon Source in the Argonne National Labora-
tory on a Mar-Research 165-mm CCD detector. These data
were processed using the HKL program. The structure was
solved and refined to 1.72 Å using CCP4/XPLOR. The stromel-
ysin-24 complex structure has been deposited with the Protein
Data Bank.24
1
H NMR: (DMSO-d6,300 mHz) δ 0.83-0.99 (m, 4H), 1.46 (dd,
J ) 15.4, 7.7 Hz, 2H), 1.71-1.77 (m, 1H), 2.38-2.45 (m, 1H),
3.50 (d, J ) 8.2, 2H), 3.77 (s, 2H), 3.85 (m, 2H), 4.07-4.11 (m,
3H), 5.16-5.25 (m, 2H), 5.72-5.80 (m, 1H), 7.13 (dd, J ) 9.0,
2.0 Hz, 2H), 7.76 (dd, J ) 9.0, 1.8 Hz, 2H). ESI MS: m/z (rel
intensity) 481.2 ([M + H]+, 100), 498.2 ([M + NH4]+, 60).
Analysis: C, H, N for C21H28N4O7S.
1N-(4-(2-Meth oxyeth oxy)ben zen esu lfon yl)-2(R)-N-h y-
d r oxyca r b oxa m id o-4(S)-1N-(3N-a llylh yd a n t oin -1N-yl)-
p yr r olid in e (73). The compound was prepared according to
the synthetic sequence described for compound 61 to give a
white powder. 1H NMR: (DMSO-d6, 300 MHz) δ 1.86 (dd, J )
11.4, 7.3 Hz, 1H), 2.44 (ddd, J ) 10.1, 10.1, 9.3 Hz, 1H), 3.34
(s, 3H), 3.48 (d, J ) 9.2 Hz, 1H), 3.54 (d, J ) 9.3 Hz, 1H),
3.68-3.74 (m, 2H), 3.77 (s, 2H), 3.85 (br d, J ) 5.7 Hz, 2H),
4.10 (d, J ) 9.0 Hz, 1H), 4.23 (dd, J ) 4.4, 4.2 Hz, 2H), 4.72-
4.84 (m, 1H), 5.15-5.27 (m, 2H), 5.69-5.83 (m, 1H), 7.17 (d,
J ) 8.8 Hz, 2H), 7.77 (d, J ) 9.0 Hz, 2H), 9.05 (br s, 1H), 10.82
(br s, 1H). ESI MS: m/z (rel intensity) 500.1 ([M + NH4]+, 15),
483.1 ([M + H]+, 100), 391.2 (20). Analysis: C, H, N for
C
20H28N4O8S‚0.5H2O.
1N-(4-n -Bu t oxyb en zen esu lfon yl)-2(R)-N-h yd r oxyca r -
Iod oa ceta te-In d u ced Ar th r itis Mod el. Sprague-Dawley
male rats weighing 220-230 g (Harlan, Indianapolis, IN) were
housed singly in wire cages in sanitary ventilated animal
rooms with controlled temperature, humidity and regular light
cycles. Rodent chow (Ralston-Purina, Richmond, IN) and water
were allowed ad libitum. Animals were acclimated for 1 week
before use.
Arthritis was induced by a single intraacticular injection of
iodoacetate into the knee joint of rats anesthetized using 3:1
CO2/O2. A 10 mg/mL concentration of monosodium iodoacetate
(MIA) (Aldrich Chemical Co., Milwaukee, WI) was prepared
using injectable saline as the vehicle. After appropriate
anesthesia, each rat was positioned on their back and the left
leg was flexed 90° at the knee. The patellar ligament was
palpated below the patella and the injection was made into
this region. Each rat received 0.25 mL intraarticular injection
into the left knee using a glass gastight syringe with a 27
gauge 0.5-in. needle. Care was taken not to advance the needle
in too far into the cruciate ligaments.
Animals were dosed with the inhibitor twice daily at 12-h
intervals (b.i.d.) for the first 7 days after iodoacetate injection.
Vehicle control and MMP inhibitor treated groups consisted
of 15 animals each. Animals were sacrificed 21 days after
iodoacetate injection and the left knee joint was immediately
disarticulated and fixed in 10% buffered formalin for 24-48
h prior to capturing the image.
After fixation, an image of the tibial cartilage surface was
captured using an Optimas image analysis system (Optimas,
Media Cybernetics LP, Silver Spring, MA). The tibial plateau
was utilized for image analysis because it provided a relatively
flat surface compared to the femoral condyles allowing the
image analysis camera to focus on the entire cartilage surface.
The image was used for grading the severity of damage.
Cartilage damage was assessed by three independent observ-
ers in a blinded manner using a scale of 0-4 of increasing
severity (0 ) normal; 4 ) maximum severity).
boxa m id o-4(S)-1N-(3N-n -p r op ylh yd a n toin -1N-yl)p yr r ol-
id in e (74). The compound was prepared according to the
synthetic sequence described for compound 61. The product
was recrystallized from cold methanol to give a white powder.
1H NMR: (DMSO-d6, 300 mHz) δ 0.81 (t, J ) 7.3 Hz, 3H),
0.95 (t, J ) 7.4 Hz, 3H), 1.39-1.53 (m, 4H), 1.69-1.80 (m, 2H),
1.85 (dd, J ) 12.4, 7.1 Hz, 1H), 2.38-2.47 (m, 1H), 3.15 (t, J
) 7.1 Hz, 2H), 3.43 (d, J ) 9.2 Hz, 1H), 3.51 (d, J ) 8.8 Hz,
1H), 3.82 (s, 2H), 4.09 (t, J ) 5.7 Hz, 2H), 4.50-4.83 (m, 1H),
7.14 (br d, J ) 8.2 Hz, 2H), 7.76 (br d, J ) 8.4 Hz, 2H), 9.04
(br s, 1H), 10.81 (br s, 1H). ESI MS: m/z (rel intensity) 483.1
([M + H]+, 100), 500.1 ([M + NH4]+, 60). Analysis: C, H, N
for C21H30N4O7S.
1N-(4-(2-Meth oxyeth oxy)ben zen esu lfon yl)-2(R)-N-h y-
d r oxyca r boxa m id o-4(S)-1N-(3N-n -p r op ylh yd a n toin -1N-
yl)p yr r olid in e (75). The compound was prepared according
to the synthetic sequence described for compound 61 to give a
1
white powder. H NMR: (DMSO-d6, 300 MHz) δ 0.83 (t, J )
7.4 Hz, 3H), 1.39-1.53 (m, 2H), 1.86 (dd, J ) 12.6, 7.7 Hz,
1H), 2.45 (ddd, J ) 11.7, 10.2, 10.2 Hz, 1H), 3.15 (t, J ) 7.1
Hz, 2H), 3.33 (s, 3H), 3.50 (d, J ) 8.4 Hz, 2H), 3.71 (dd, J )
4.8, 3.7 Hz, 2H), 3.83 (s, 2H), 4.10 (d, J ) 8.8 Hz, 1H), 4.23
(dd, J ) 4.9, 3.3 Hz, 2H), 4.71-4.84 (m, 1H0, 7.17 (br d,
J ) 8.8 Hz, 2H), 7.77 (br d, J ) 8.6 Hz, 2H), 9.06 (br s, 1H),
10.77 (br s, 1H). ESI MS: m/z (rel intensity) 485.0 ([M + H]+,
100), 502.0 (M + NH4]+, 10). Analysis: C, H, N for C20H28
N4O8S.
-
MMP In h ib it ion Assa y. The preparation of the human
recombinant MMPs used in these studies has been described
previously.17 MMP inhibitors were tested for their ability to
inhibit human MMPs using the quenched fluorescence assay.21
This assay was modified to fit a 96-well format to increase
the throughput. Assays for MMP-1, -3, -7, -8 and -13 employed
human recombinant truncated enzymes. The optimal amount
of each enzyme to produce significant and reproducible sub-
strate cleavage was determined in preliminary experiments.
Assays for MMP-2 and -9 utilized human recombinant full
length enzymes. MMP-2 was activated by incubating proM-
MP-2 with APMA for 45 min. The final concentration of
MMP-2 in the assay mixture was 1 nM. ProMMP-9 was
activated with MMP-3 (ratio 20:1) for 2 h and diluted to a final
assay concentration of 0.75 nM. The final concentration of
Sta tistica l An a lysis. Data were analyzed using a non-
parametric procedure (Wilcoxon rank sum). The data are
expressed as the mean of inhibition of joint damage relative
to the vehicle-treated control group. Statistical differences from
the vehicle treated control (p < 0.05) are denoted with an
asterisk.

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