N-i-Propoxy-N-biphenylsulfonylaminobutylhydroxamic acids as potent and selective inhibitors of MMP-2 and MT1-MMP

Article abstract of DOI:10.1016/j.bmcl.2005.01.024

Structural manipulation of the pharmacophoric model of type A selective MMP inhibitors (MMPi), obtained by the insertion of some alkyl substituents R ₂ possessing an appropriate geometry, steric bulkiness and lipophilicity, is able to improve p

Full text of DOI:10.1016/j.bmcl.2005.01.024

Bioorganic & Medicinal Chemistry Letters 15 (2005) 1321–1326
N-i-Propoxy-N-biphenylsulfonylaminobutylhydroxamic acids
as potent and selective inhibitors of MMP-2 and MT1-MMP
Armando Rossello,a,* Elisa Nuti,a Paolo Carelli,a Elisabetta Orlandini,a Marco Macchia,a
Susanna Nencetti,a Maurizio Zandomeneghi,b Federica Balzano,b
Gloria Uccello Barretta,b Adriana Albini,c Roberto Benelli,c Giovanni Cercignani,d
Gillian Murphye and Aldo Balsamoa
a
`
Dipartimento di Scienze Farmaceutiche, Universita degli Studi di Pisa, Via Bonanno, 6, 56126 Pisa, Italy
`
b
Dipartimento di Chimica e Chimica Industriale, Universita degli Studi di Pisa, Via Risorgimento, 35, 56126 Pisa, Italy
cIstituto Nazionale per la Ricerca sul Cancro, Via Rosanna Benzi 10, 16132 Genova, Italy
d
`
Dipartimento di Fisiologia e Biochimica, Universita degli Studi di Pisa, Via San Zeno, 51, 56127 Pisa, Italy
eDepartment of Oncology, University of Cambridge, Hills Road, Cambridge CB2 2XY, UK
Received 13 October 2004; revised 10 January 2005; accepted 12 January 2005
Abstract—Structural manipulation of the pharmacophoric model of type A selective MMP inhibitors (MMPi), obtained by the
insertion of some alkyl substituents R2 possessing an appropriate geometry, steric bulkiness and lipophilicity, is able to improve
potency, in the subnanomolar range on MMP-2, and to give a good MMP inhibition on MMP-14 (MT1-MMP) in the designed
MMPi of type C, while maintaining a good MMP-1/MMP-2 selectivity profile. The simultaneous inhibition of these two enzymes
yields type C compounds, which are potent antiangiogenic agents, able to block a chemoinvasion model on HUVEC cells in the
micromolar range.
Ó 2005 Elsevier Ltd. All rights reserved.
Often during tumour progression, an overexpression of
matrix metallo proteinases (MMPs), produced and se-
creted as inactive zymogens in the extracellular matrix
(ECM) by the tumour cells themselves, or by surround-
ing stromal cells, is responsible for the deregulation of
the extracellular matrix (ECM) functions.1,2 Normally,
the homeostasis of MMPs is maintained by tissue inhibi-
tors, TIMPs, but in cancer progression, control over
MMPs activity is lost.3 Other proteases, such as uPa,
MMPs and furin-like serine proteases are responsible
for the activation of specific MMPs, such as MMP-2,
MMP-9, and MMP-14 (a membrane-associated type
MMP which is the principal activator of pro-MMP-2).
These three specific MMPs play a significant role in
these degenerative processes, and are directly involved
in metastatic tumour dispersion and angiogenesis.4–8
More recently, MMP-2 has been shown to play other
important roles in tumours, for example, in resistance
to apoptosis, in the activation of EGF receptors and
in cellular prolification.9–12 All these data, taken to-
gether, indicate that MMP-2, MMP-9 and even MMP-
14 may represent an important target to develop new
potential anticancer drugs.13,14 In the past few years,
some potent Ôbroad spectrumÕ MMPi have been pro-
posed and tested against tumours, but at present none
of them are on the market.15,16
In fact, many of these new molecules have shown a se-
vere musculoskeletal syndrome, with fibroproliferative
effects in the joint capsule of the knee.17–19 These effects
are thought to be linked to an impairment of normal tis-
sue remodelling governed by MMP-1 and/or by shedd-
ases such as TNF-a-convertase.20 For these reasons, a
lack of activity with respect to MMP-1 is considered
to be an important factor in reducing some of the side
effects found for Ônon-selectiveÕ MMPi.21 As a matter
of fact, the recent development of some synthetic MMPi
possessing a good potency and selectivity towards the
Keywords: MMP-inhibitors; MMP-2/MT1-MMP selective inhibitors;
Antiangiogenic agents.
*
Corresponding author. Tel.: +39 050 2219562; fax: +39 050
2219605; e-mail: aros@farm.unipi.it
0960-894X/$ - see front matter Ó 2005 Elsevier Ltd. All rights reserved.
doi:10.1016/j.bmcl.2005.01.024

1322
A. Rossello et al. / Bioorg. Med. Chem. Lett. 15 (2005) 1321–1326
two gelatinases, together with the discovery that some of
these molecules active on MMP-2 show important pro-
apoptotic effects on tumour cell cultures, confirmed the
validity of their use as potential anti-tumour agents.22–29
Nowadays the development of studies on compounds
possessing a selective inhibitory activity on the MMPs
that are over-expressed in tumours is very useful, with
particular reference to viability control and invasiveness
of the cancer cell.15
hydroxamic group of A, thinking that these substituents,
if appropriately oriented, might favour lipophilic inter-
actions with the nearby S1 region of the enzyme site,
thus further reinforcing the binding of the planned
new inhibitors to the active site of MMP-2 and MMP-
14. By introducing an asymmetric centre into molecules
C, this kind of chemical manipulation might allow us to
evaluate the enantioselective recognition of the studied
MMPs towards this class of compounds and, therefore,
the influence of chirality on their biopharmacological
properties. According to our hypothesis, in consider-
ation of the differences existing in the S1, S10 and S20
pockets between MMP-1, MMP-2 and MMP-14, the
combination of appropriate molecular geometries, hin-
drances and lipophilic characteristics of the groups R,
R1 and R2, introduced into type C compounds might
be useful to improve their MMPi potency and selectiv-
ity.36 Furthermore, the new compounds C may be able
to give an antiangiogenic activity, seeing that they act
as potent inhibitors versus MMP-2 and MMP-14, the
two MMPs over-expressed in tumours characterised by
particular aggressiveness, sparing the inhibitory activity
on MMP-1 responsible for undesirable serious side
effects.
In a recent work, we described some N-arylsulfonyl-N-
alkoxyaminoacetohydroxamic acids of type A. Some
of the new compounds, when tested in vitro for their
inhibitory activity towards some MMPs, showed a
remarkable activity and an already good MMP2 selec-
tivity, with IC50s in the nanomolar range. Moreover
the potent MMP-2 inhibitor 1, belonging to the class
A, proved to possess an appreciable anti-invasive effi-
cacy.30 In an aim to improve the potency of these mole-
cules towards MMP-2 and MMP-9, and also towards
MMP14, where type A compounds have proved to be
devoid of any activity (see compound 1 in Table 1), we
planned a chemical manipulation of this new type of
model of pharmacophore in the class of MMPi. Our
interest in the study of these inhibitors also on the
MMP-14, which is significantly expressed at the mem-
brane level in tumour cells, is linked to the particular
role of this MMP in cancer cells. MMP-14, together
with TIMP-2, the natural tissue inhibitor of MMP-2,
closely controls the activation/inactivation balance of
pro-MMP-2, and therefore it may be directly involved
in the crucial regulation of the matrix functions around
the metastatic cells in invasive tumours, thereby greatly
facilitating motility, invasion, metastasis and angiogene-
sis (see Fig. 1).31–35 On this basis, we have developed
some new N-i-propoxy-N-biphenylsulfonylaminobu-
tylhydroxamic acids of type C, which are analogues of
the previously studied sulfonamide 1 of type A, in order
to evaluate their biopharmacological properties. In these
new type C hydroxamates, we have introduced some al-
kyl substituents possessing an increasing steric bulkiness
and lipophilicity in R2, on the carbon atom alpha to the
The synthesis of the new type C derivatives is outlined in
Schemes 1 and 2. Firstly, in the aim to improve our
knowledge on the stereochemical aspects of type C com-
pounds action, we planned the synthesis of racemic (R/
S)-5a and of its enantiomers (R)-5a and (S)-5a then,
identified the best stereochemistry on the alpha carbon
atom to the hydroxamate binding group, some new type
C derivatives of R configuration were synthesised.
Scheme 1 reports the synthesis of racemic (R/S)-5a and
optically active (S)-5a according to the synthetic method
used for type A hydroxamates.30 In the same scheme,
there is an attempt to obtain (R)-5a, using the same syn-
thetic method, by nucleophilic SN2 replacement reaction
of optically active secondary bromides with a cesium salt
of the O-i-propyl-sulfonamide 6. Unfortunately, only
Ph
i,ii
Ph
O
O
Ph
O
O
R1
R1
O
O
O
O
S
O
S
N
O
O
O
S
N
v
O
S
N
O
S
N
HN
O
HO
O
HO
HO
O
PEA
N
H
O
R
N
H
C
R
H2
R2
6
(R/S)-7a
(S,S)-9a, (R,S)-9a
(S)-7a, (R)-7a
A
B
iii
3, CGS27023A, R = 3-Py, R1 = OMe, R2 = i-Pr
4, AG-3340, c-(R-R2) = -(Me)2CS(CH2)-, R1 = OPh
1, R = i-Pr, R1 = Ph
2, R = i-Pr, R1 = MeO
Ph
O
O
Ph
O
O
O
S
iv
O
S
N
R1
O
O
TBDMSO
N
O
HO
N
H
O
O
N
H
O
O
S
N
O
S
N
HO
α
HO
N
H
O
R
(R/S)-8a
(S)-8a
N
H
O
(R/S)-5a
(S)-5a
R2
C
R2
PEA = (S)-1-Phenylethylamina
Scheme 1. Reagents and conditions: (i) Racemic or (R)- or (S)-
BrCH[CH(CH3)2]CO2–But, Cs2CO3, Bu4NHSO4, anhyd DMF, rt,
3 days; (ii) TFA, anhyd DCM, 0 °C; (iii) TBDMSONH2, EDCI, anhyd
DCM, 0–25 °C, 24 h; (iv) TFA, anhyd DCM 0 °C, 5 h; (v) (S)-1-PEA,
NMM, HOBt, EDCI, anhyd DCM, rt, 12 h.
(R)-5a; (S)-5a
(R)-5b; (R)-5c
a, R2 = i- Pr; b, R2 = CH2CH2NHCBz; c, R2 = CH2CH2NH2
Figure 1.

A. Rossello et al. / Bioorg. Med. Chem. Lett. 15 (2005) 1321–1326
1323
O
O
O
silylate (R)-15a and (R)-15b in the same conditions de-
scribed above gave the desired hydroxamates (R)-5a
and (R)-5b. In the case of the compound (R)-5c
(R2 = CH2CH2NH2), the synthetic procedure was modi-
fied slightly because it is not possible to obtain (R)-5c by
direct catalytic hydrogenation of its hydroxamate pre-
cursor (R)-15b. In this case, the acid (R)-14b was con-
verted into its O-benzyl-hydroxamate (R)-16,and then
the desired (R)-5c was obtained by catalytic hydrogena-
tion on 10% Pd/C.39,40
i
ii,ii
OH
OAc
OH
HO
HO
O
R2
(S)-10a, (S)-10b
R2
(S)-11
(S)-12a, (S)-12b
ii
+
Ph
O
O
Ph
O
S
O
S
N
O
i
O
O
HN
O
R2
(R)-13a,
(R)-13b
Table 1 shows the inhibitory indices (IC50) towards
some of the principal MMPs of the new N-i-propoxy-
N-biphenylsulfonylaminobutylhydroxamic acids of type
C (5a-c), compared with those of the previously de-
scribed type A MMPi 1 (R = i-Pr; R1 = Ph). In the same
table, the IC50 values are reported for two related
known MMPi of type B, CGS27023A (3) and AG-
3340 (4).
6
v
Ph
O
S
Ph
O
O
O
O
O
S
N
N
O
vii
BnOHN
HO
O
R2
(R)-14a,
(R)-14b
NHCBz
(R)-1
For the more interesting compounds and reference
drugs, selectivity indices for MMP-2 over the other
MMPs studied are also reported, expressed as ratios of
their inhibitory indices (Table 1).
i
v
Ph
O
S
Ph
O
O
O
O
O
S
N
HO
N
O
vi
TBDMSO
N
H
On MMP-2, the R2 substitution on the P1 site with an i-
Pr group on the carbon atom alpha to the hydroxamate
is able to improve the inhibitory potency about 7-fold,
passing from 1 (IC50 = 12 nM) to racemic (R,S)-5a
(IC50 = 1.8 nM). The (R)-5a enantiomer shows an
improvement of the potency on MMP-2 of about 20-
fold (IC50 = 0.09 nM) compared with its racemate
(R,S)-5a and about 114 times, compared with its
dystomer (S)-5a (IC50 = 10.3 nM). On MMP-9, the
same racemate (R,S)-5a (IC50 = 19.6 nM) shows an
appreciable increase in potency of about 10 times com-
pared with the type A compound 1 (IC50 = 200 nM). On
the same enzyme, the eutomer (R)-5a shows an
IC50 = 6.7 nM, about 3 times that of (R,S)-5a and about
twice that of its enantiomer (S)-5a (IC50 = 13 nM). (R)-
5a shows good indices of potency also on MMP-3
(IC50 = 50 nM), and on MMP-8 where the inhibitory
activity of this compound (IC50 = 1.6 nM) is particu-
larly significant.
N
H
O
R2
R2
(R)-15a,
(R)-5a, (R)-5b, (R)-5c
a, R2 = i- Pr; b, R2 = CH2CH2NHCBz; c, R2 = CH2CH2NH2
Scheme 2. Reagents and conditions: (i) AcCl, rt, 18 h; (ii)
(CH3)2NCH[O(CH3)3]2, toluene, 95 °C, 3 h; (iii) LiOH, THF/MeOH/
H2O, rt, 3 h; (iv) TPP, DEAD, anhyd THF, 0 °C, 5 h; (v) TFA, anhyd
DCM, 0 °C, 5 h; (vi) TBDMSiONH2, EDCI, anhyd DCM, 0–25 °C,
24 h; (vii) TFA, anhyd DCM 0 °C, 5 h; (viii) BnONH2 HCl, NMM,
HOBt, EDCI, anhyd DCM, rt, 24 h; (ix) H2, Pd/C 10%, MeOH/
AcOH.
one of the two desired enantiomeric acids, (S)-7a, was
obtained with an acceptable enantiomeric purity.37 Only
acid (S)-7a was therefore transformed into its hydrox-
amate (S)-5a in accordance with the previously de-
scribed method. The absolute configuration of (S)-5a
was determined by comparison of its optical characteris-
tics with an authentic sample of its enantiomer (R)-5a
prepared and optically characterised as described below.
The enantiopure biphenylsulfonamides of type C with
the desired R configuration, (R)-5a, (R)-5b, and (R)-5c,
were synthesised as described in Scheme 2. Optically ac-
tive alpha-hydroxy-tert-butyl-ester, (S)-12a (R2 = i-Pr)
was synthesised by acetylation of the commercial al-
pha-hydroxy acid (S)-10a to the stable intermediate
(S)-11a (R2 = i-Pr) and subsequent esterification to (S)-
12a (R2 = i-Pr).38 On the contrary, the ester (S)-12b
was prepared directly from the alpha-hydroxy acid (S)-
10b (R2 = CH2CH2NHCBz) by direct esterification with
N-(di-tert-butoxymethyl)-N,N-dimethylamine. A Mits-
unobu condensation of the O-i-propyl-sulfonamide 6
with the appropriate alpha-hydroxy-tert-butyl-ester,
(S)-12a and (S)-12b, gave tert-butyl esters (R)-13a and
(R)-13b. Acid cleavage of esters 13 gave acids (R)-14a
and (R)-14b, which were converted into their O-silylates
(R)-15a and (R)-15b. Acid cleavage of the tert-butylO-
The R2 substitution on the P1 site with a more hindered
group, such as in the benzylethylcarbamate, compound
(R)-5b, reduces the inhibitory activity on MMP-2 about
4.5-fold, even if this remains in the subnanomolar range
(IC50 = 0.41 nM), compared with that of its R2 i-Pr
substituted analogue (R)-5a. More marked for this com-
pound, (R)-5b, is the loss of inhibitory activity against
MMP-1, where (R)-5b is 20 times less active
(IC50 > 3000 nM versus IC50 = 147 nM for the parent
compound (R)-5a).
On MMP-9, (R)-5b shows an IC50 = 16 nM, 2.4 times
less that of its most potent analogue (R)-5a.
The best results obtained, with these new type C com-
pound, are those found for the inhibition of MMP-14
where (R)-5a and (R)-5b, display IC50 values in the
nanomolar range. Compared with the type A compound

1324
A. Rossello et al. / Bioorg. Med. Chem. Lett. 15 (2005) 1321–1326
Table 1. Inhibitory activity of new type C compounds compared with the type A inhibitor 1 and the reference drugs 3 and 4 towards some MMPs.
MMP-2 selectivity is shown, in parentheses, for the more active compoundsa
Compd
R2
H
IC50 (nM)30
MMP-7
MMP-1
MMP-2
12
MMP-3
MMP-8
MMP-9
MMP-14
1
>50,000
(>4000)a
4500
(375)a
>50,000
(>4000)a
200
>10,000
(>833)
(17)a
19.6
6.7
(R/S)-5a
(R)-5a
i-Pr
i-Pr
1.8
0.09
147
(1633)a
50
(556)a
>1000
(>11,111)
1.6
(18)a
9.8
(109)a
(74)a
(S)-5a
(R)-5b
i-Pr
CH2CH2 NHCbz
10.3
0.41
13
>3000
(>7300)a
130
(317)a
>10,000
(>24,390)
16
7.7
(39)a
4517
8
(19)a
>300
8.9
(R)-5c
3, CGS27023A
CH2CH2NH2
935
20
55
22
100
(5)a
54
(600)a
(2.8)a
8.2
(1.1)a
0.23
(2.5)a
(0.4)a
0.26
(2.9)a
(0.4)a
0.3
(3.3)a
4, AG-3340
0.09
(91)a
a Selectivity for MMP-2 over each of the other MMPs, is expressed as the ratio of the IC50 value for MMPn over the value for MMP-2.30
1, where the IC50 > 10,000 nM, the two MMPi, (R)-5a
and (R)-5b, show an inhibitory potency that is more
than 1000 times higher (IC50 = 9.8 nM for (R)-5a and
IC50 = 7.7 nM for (R)-5b). Finally, the removal of the
CBz group on the R2 substituent of (R)-5b to obtain
the amino derivative (R)-5c caused a further loss of
inhibitory activity of 2280-fold on MMP-2 (IC50
=
935 nM) and of 282-fold on MMP-9 (IC50 = 4517 nM).
Probably the presence of a protonable nitrogen as in
(R)-5c, is not accepted in this site.
An analysis of the selectivity indices reported in paren-
theses in Table 1 for the N-i-propoxy-N-biphenyl-
sulfonylaminobutylhydroxamic acids of type A.
(1), C (5a–c) and the reference drugs 3 and 4 indicates
that the newly synthesised type C compounds (R)-5a
and (R)-5b show a high selectivity profile (MMP-1/
MMP-2 ratio of 1633 for (R)-5a and >7300 for (R)-
5b). These two potent MMPi, (R)-5a and (R)-5b, show
good selectivity indices versus MMP-3, with an MMP-
2/MMP-3 ratio of 556 for (R)-5a and 317 for (R)-5b,
and versus MMP-7, with an MMP-2/MMP-7 ratio
>11,111 for (R)-5a and >24,390 for (R)-5b. Selectivity
indices are lower on MMP-8 (MMP-2/MMP-8 ratio of
18 for (R)-5a), on MMP-9 (MMP-2/MMP-9 ratio of
74 for (R)-5a and39 for (R)-5b) and on MMP-14
(MMP-2/MMP-9 ratio of 109 for (R)-5a and 19 for
(R)-5b). A comparison of the selectivity indices with
the previously studied N-4-biphenylsulfonyl-N-i-prop-
oxyaminoacetohydroxamic acid 1 of type A indicates
an improvement of the selectivity ratio MMP-1/MMP-
2 in the most potent new MMP-2 inhibitor (R)-5b of al-
most 2-fold. The new inhibitor (R)-5a is certainly more
potent on MMP-2 than the reference drug 3, and is as
potent, on the same enzyme, as AG-3340 (4). On
MMP-14, the two new inhibitors show a similar inhibi-
tory potency to those of the known sulfonamides chosen
as reference drugs.
Figure 2. HUVEC–Chemoinvasion Test: Negative control (column 1)
represents the background random migration of HUVEC in the
presence of serum free, unconditioned medium (SFM). Positive control
(column 2) represents the maximal chemotactic response of HUVEC in
the presence of 3T3-CM. Invasion control (column 3) represents
the maximal ability of HUVEC to cross the matrigel barrier in the
presence of 3T3-CM. Columns 4, 5, 6 and 7 represent, respectively, the
ability of HUVEC to cross the matrigel barrier in the presence of
the (R)-5a inhibitor 1 and 0.1 lM (columns 4 and 5) and of the (R)-5b
inhibitor at the same concentrations (columns 6 and 7), in the presence
of 3T3-CM.
The two potent and selective type C MMPi synthesised,
(R)-5a and (R)-5b, were evaluated in the chemoinvasion
assay, an in vitro model of angiogenesis, able to quantify
the invasive potential of endothelial cells. Figure 2 re-
ports the results of this test, using human umbilical vein
endothelial cells (HUVEC), stimulated to invade a
reconstituted basement membrane (matrigel) by 3T3
fibroblast-derived conditioned medium (3T3-CM).

A. Rossello et al. / Bioorg. Med. Chem. Lett. 15 (2005) 1321–1326
1325
15. Coussens, L. M.; Fingleton, B.; Matrisian, L. Science
2002, 295, 2387.
16. Borkakoti, N. Biochem. Soc., Trans. 2004, 32, 17–
19.
17. Hutchinson, J. W.; Tierney, G. M.; Parson, S. L.; Davis,
T. R. C. J. Bone Joint Surgery 1998, 80, 907.
18. Holmbeck, K.; Bianco, P.; Caterina, J.; Yamada, S.;
Kromer, M.; Kuznetsov, S. A.; Mankani, M.; Robey, P.
G.; Poole, A. R.; Pidoux, I.; Ward, J. M.; Birkedal-
Hansen, H. Cell 1999, 99, 81.
The chemoinvasion assay clearly shows that both (R)-5a
and (R)-5b are able to reduce considerably the ability of
HUVEC cells to cross the matrigel barrier. At a concen-
tration of 1 lM, both inhibitors reduced invasion to
basal level, while at 0.1 lM, the effect of (R)-5a and
(R)-5b was less impressive. On this basis, both (R)-5a
and (R)-5b inhibitors can be considered to be potent
antiangiogenic agents, able to block the chemoinvasion
of HUVEC cells in the micromolar range.41
19. Steward, W. P. Cancer Chemother. Pharmacol. 1999, 43,
S56.
20. Dahlberg, L.; Billinghurst, R. C.; Manner, P.; Nelson, F.;
Webb, G.; Ionescu, M.; Reiner, A.; Tanzer, M.; Zukor,
D.; Chen, J.; Van Wart, H. E.; Poole, A. R. Arthritis
Rheum. 2000, 43, 673.
21. Scatena, R. Exp. Opin. Invest. Drugs 2000, 9, 2159.
22. Scozzafava, A.; Owa, T.; Mastrolorenzo, A.; Supuran, C.
T. Curr. Med. Chem. 2003, 10, 925.
23. Wada, C. K.; Holms, J. H.; Curtin, M. L.; Dai, Y.;
FlorJancic, A. S.; Garland, R. B.; Guo, Y.; Heyman, H.
R.; Stacey, J. R.; Steinman, D. H.; Albert, D. H.; Bouska,
J. J.; Elmore, I. N.; Goodfellow, C. L.; Marcotte, P. A.;
Tapang, P.; Morgan, D. W.; Michaelides, M. R.; David-
sen, S. K. J. Med. Chem. 2002, 45, 219.
24. Supuran, C. T.; Casini, A.; Scozzafava, A. Med. Res. Rev.
2003, 23, 535.
25. Zook, S. E.; Dagnino Jr., R.; Deason, M. E.; Bender, S.
L.; Melnick, M. J. WO Patent 97/20824; 1997, 127,
108945.Chem. Abstr. 1995, 123, 2870.
26. Supuran, C. T.; Scozzafava, A. In Proteinase and Pepti-
dase Inhibitors: Recent Potential Targets for Drug Devel-
opment; Smith, H. J., Simons, C., Eds.; Taylor & Francis:
London & New York, 2002; pp 35–61.
27. Bernardo, M. M.; Brown, S.; Li, Zhi-Hong; Fridman, R.;
Mobashery, S. J. Biol. Chem. 2002, 277, 11201.
28. Rabbani, S. A.; Harakidas, P.; Guo, Y.; Steinman, D.;
Davidsen, S. K.; Morgan, D. W. Int. J. Cancer 2000, 87,
276.
In conclusion, in the new N-i-propoxy-N-biphen-
ylsulfonylaminobutylhydroxamic acids of type C, de-
signed as analogues of type A inhibitors, the R2 alkyl
substituents introduced on the carbon atom alpha to
the hydroxamic group of the pharmacophoric portion
of A seem to confirm our starting hypothesis: this new
R2 substitution site may strongly favour lipophilic inter-
actions with the nearby S1 region of the enzyme site,
reinforcing the binding of the planned new inhibitors
to the active site of MMP-2. Moreover, in the case of
the targeted MMP-14, this chemical manipulation of
the type A pharmacophore increases the MMPi potency
on this crucial MMP more than 1000-fold, shifting their
IC50 values from the micromolar to the nanomolar
range. A similar structural modification on the pharma-
cophoric portion of MMPi of type A does not substan-
tially modify the MMP-1/MMP-2 selectivity profile, but
maintains or reinforces the good indices. Finally, the
new type C MMPi, which prove to be active against
these two MMPs, MMP-2 and MMP-14, are potent
antiangiogenic agents, able to block the chemoinvasion
of HUVEC cells in the micromolar range.
References and notes
29. Nyormoi, O.; Mills, L.; Bar-Eli, M. Cell Death Differ.
2003, 5, 558.
30. Rossello, A.; Nuti, E.; Orlandini, E.; Carelli, P.; Rappos-
elli, S.; Macchia, M.; Minutolo, F.; Carbonaro, L.; Albini,
A.; Benelli, R.; Cercignani, G.; Murphy, G.; Balsamo, A.
Bioorg. Med. Chem. 2004, 12, 2441.
1. Seiki, M.; Yana, I. Cancer Sci. 2003, 94, 569–574.
2. Folguera, A. R.; Pendas, A. M.; Sanchez, L. M.; Lopez-
Otin, C. Int. J. Dev. Biol. 2004, 48, 411–424.
3. Baker, A. H.; Edwards, D. R.; Murphy, G. J. Cell Sci.
2002, 115, 3719.
4. Murphy, G.; Crabbe, T. Methods Enzymol. 1995, 248,
470.
5. Kleiner, D. E.; Stetler-Stevenson, W. G. Cancer Chemo-
ther. Pharmacol. 1999, 43, S42.
6. Aimes, R. T.; Quigley, J. Q. J. Biol. Chem. 1995, 270,
5872.
31. Sato, H.; Takino, T.; Okada, Y.; Cao, J.; Shinagawa, A.;
Yamamoto, E.; Seiki, M. Nature 1994, 370, 61.
32. Butler, G. S.; Butler, M. J.; Atkinson, S. J.; Will, H.;
Tamura, T.; Van Westrum, S. S.; Crabbe, T.; Clements, J.;
DÕorto, M. P.; Murphy, J. J. Biol. Chem. 1998, 273,
871.
7. Itoh, T.; Tanioka, M.; Matsuda, H.; Nishimoto, H.;
Yoshioka, T.; Suzuki, R.; Uehira, M. Clin. Exp. Metastas.
1999, 17, 177.
8. Lafleur, M. A.; Forsyth, P. A.; Atkinson, S. J.; Murphy,
G.; Edwards, D. R. Biochem. Biophys. Res. Commun.
2001, 282, 463.
33. Maquoi, E.; Frankenne, F.; Barmova, E.; Munaut, C.;
Sounni, N. E.; Remacle, A.; Noel, A.; Murphy, G.;
Foidart, J. M. J. Biol. Chem. 2000, 275, 11368.
34. Overall, C. M.; Tam, E.; McQuibban, G. A.; Morrison,
C.; Wallon, U. M.; Bigg, H. F.; King, A. E.; Roberts, C.
R. J. Biol. Chem. 2000, 275, 39497.
35. Sounni, N. E.; Roghi, C.; Chabottaux, V.; Janssen, M.;
Munaut, C.; Maquoi, E.; Galvez, B. G.; Gilles, C.;
Frankenne, F.; Murphy, G.; Foidart, J.-M.; Noel, A. J.
Biol. Chem. 2004, 279, 13564.
9. Cowan, K. N.; Jones, P. L.; Rabinovitch, M. Circ. Res.
1999, 84, 1223.
10. Jones, P. L.; Crack, J.; Rabinovitch, M. J. Cell Block
1997, 279.
36. Terp, G. E.; Cruciani, G.; Christensen, I. T.; Jørgensen, F.
S. J. Med. Chem. 2002, 45, 2675.
11. Eguchi, P. J.; Dempsey, G. D.; Frank, G. D.; Motley, D.;
Inagami, T. J. Biol. Chem. 2001, 276, 7957.
12. Ahonen, M.; Poukkula, M.; Baker, A. H.; Kashiwagi, M.;
Nagase, H.; Eriksson, J. E.; Ka¨ha¨ri, V.-M. Oncogene 2003,
22, 2121.
13. Giannelli, G.; Antonaci, S. Histol. Histopathol. 2002, 17,
339.
14. John, A.; Tuszynski, G. Pathol. Oncol. Res. 2001, 7, 14.
37. Using a conventional method to evaluate the optical
purity of carboxylic acids, the obtained enriched acids (S)-
7a and (R)-7a were transformed into their diastereoiso-
meric amides (S,S)-9a and (R,S)-9a with (S)-1-phenyleth-
ylamine ((S)-PEA). The resulted diastereoisomeric amides,
(S,S)-9a and (R,S)-9a, subjected to HPLC analysis to

1326
A. Rossello et al. / Bioorg. Med. Chem. Lett. 15 (2005) 1321–1326
establish their optical purity, gave as results an acceptable
ee of 88% only for the corresponding acid (S)-7a.
38. Dan, Y.; Bing, L.; Fei-Fu, N.; Yi-Long, Y.; Jin, Q.; Yun-
39. Rosini, C.; Uccello-Barretta, G.; Pini, D.; Abete, C.;
Salvadori, P. J. Org. Chem. 1988, 53, 4579.
40. Hanessian, H.; Yang, R.-Y. Synlett 1995, 633.
41. Benelli, R.; Albini, A. Int. J. Biol. Markers 1999, 14, 243–246.
Dong, W. J. Org. Chem. 2001, 66, 7303.