Structure-activity relationships of azasugar-based MMP/ADAM inhibitors

Article abstract of DOI:10.1016/S0960-894X(03)00530-4

In order to investigate structure-activity relationships of azasugar series toward metalloproteinases, we synthesized and evaluated several azasugar-based compounds. As a result, it was found that 4-phenoxybenzene derivative 3 having 2R,3R,4R,5S-configura

Full text of DOI:10.1016/S0960-894X(03)00530-4

Bioorganic & Medicinal Chemistry Letters 13 (2003) 2737–2740
Structure–Activity Relationships of Azasugar-Based
MMP/ADAM Inhibitors
Hideki Moriyama,a,b Takahiro Tsukida,a,b,*,y Yoshimasa Inoue,b Hirosato Kondo,b,y
Kohichiro Yoshinob and Shin-Ichiro Nishimurac,*
aJapan Bioindustry Association, Hokkaido Collaboration Center, N-21, W-12, Kita-Ku, Sapporo 001-0021, Japan
bR&D Laboratories, Nippon Organon K.K., 1-5-90, Tomobuchi-cho, Miyakojima-ku, Osaka 534-0016, Japan
cDivision of Biological Sciences, Graduate School of Science, Hokkaido University, N-21, W-12, Kita-Ku, Sapporo 001-0021, Japan
Received 17 February 2003; revised 16 May 2003
Abstract—In order to investigate structure–activity relationships of azasugar series toward metalloproteinases, we synthesized and
evaluated several azasugar-based compounds. As a result, it was found that 4-phenoxybenzene derivative 3 having 2R,3R,4R,5S-
configurations exhibited most potent inhibitory activities against matrix metalloproteinase-1, -3 and -9 and TACE.
# 2003 Elsevier Ltd. All rights reserved.
Metalloproteinases, contained matrix metallo-protein-
ases (MMPs) and a disintegrin and metallo-proteinases
(ADAMs), are a family of zinc-containing enzymes.
MMPs, comprised collagenases, stromelysins, gelatin-
ases and membrane-type MMPs (MT-MMPs), mediate
the breakdown of connective tissue and are therefore
targets for therapeutic inhibitors in many inflammatory,
malignant and degenerative diseases.1À4 On the other
hands, ADAMs,5 structurally related to MMPs, med-
iate the processing of membrane-bound cytokine such
as tumor necrosis factor a (TNF-a) into soluble form.
TNF-a is a major mediator of inflammatory and
immune responses6 and a strong inducer of other cyto-
kines such as IL-lb, IL-6 and IL-8. Elevated TNF-a
levels are implicated in pathologies of rheumatoid arthri-
tis,7 multiple sclerosis,8 type II diabetes,9 and so on.
Therefore, TNF-a converting enzyme (TACE or
ADAM17) is an attractive target for medicinal chemists.10
the inhibitory activity toward MMPs and TACE by
synthesizing new analogue of compound 1a. In the
present report, we investigated the effects of stereo-
chemistry at C-3, C-4, and C-5 positions, and property
of arylsulfonylamide moiety on biological activity of
azasugar-based inhibitors.
Chemistry
Figure 1 shows a synthetic strategy of new analogues of
azasugar-based metalloproteinases inhibitors from 1a as
a key compound. Schemes 1 and 2 indicate synthetic
routes of analogues appeared in this study.
The azide compounds 6b–d,12 which were easily pre-
pared from l-gulono-1,4-lactone 6b, l-glucono-1,5-lac-
tone 6c and d-gulono-1,4-lactone 6d, respectively, were
hydrogenated in the presence of palladium on carbon,
and then the corresponding amines were reacted with 4-
methoxybenzenesulfonyl chloride 7 to give compounds
8b–d in 72-87% yields (Scheme 1). The terminal iso-
propylidene group were selectively cleaved by treatment
of Muromac1 (H+-form), in 90% MeOH13a or cerium
chloride heptahydrate and catalytic amount of oxalic
acid in acetonitrile13b to provide diols 9b–d in moderate
yields. The primary hydroxyl group of diols 9b–d were
selectively mesylated by treatment of mesyl chloride at
À40 ꢀC in dichloromethane to afford mesylates 10b–d in
moderate yields. The intramolecular cyclization of com-
pounds 10b–d in the presence of potassium carbonate
As previously reported, we succeeded to find novel aza-
sugar-based MMP/ADAM inhibitors such as com-
pound 1a,11 having 2R,3S,4R,5S-configuration, which
exhibited moderate inhibitory activity against MMP-1, -3,
-9 and TACE. Next, we focused on the optimization of
*Corresponding author. Tel.: +81-6-6401-8199; e-mail: takahiro.
tsukida@shionogi.co.jp (T. Tsukida); Tel.: +81-11-706-9043;
fax: +81-11-706-9042; e-mail: shin@glyco.sci.hokudai.ac.jp (S.I.
Nishimura).
yPresent address: Manufacturing Technology R&D Laboratories,
Shionogi & Co., Ltd., 1-3 Kuise Terajima 2-chome, Amagasaki,
Hyogo 660–0813, Japan.
0960-894X/03/$ - see front matter # 2003 Elsevier Ltd. All rights reserved.
doi:10.1016/S0960-894X(03)00530-4

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H. Moriyama et al. / Bioorg. Med. Chem. Lett. 13 (2003) 2737–2740
Figure 1. Synthetic strategy of new analogues of azasugar-based metalloproteinase inhibitor.
Scheme 1. (a) 10% Pd-C/H2, EtOAc; (b) 7, 4-DMAP, DMF, 72–87% from 6b–d; (c) Muromac, 90% MeOH; (d) cerium chloride heptahydrate,
oxalic acid, CH3CN, 53–64%; (e) methanesulfonyl chloride, Et3N, CH2Cl2, 31–58%; (f) K2CO3, DMF, 56–100%; (g) 50% NH2OH aq., NaCN,
MeOH, 51–72%; (h) Muromac, MeOH, 66–78%.
gave compounds 11b–d. Compounds 11b–d were sub-
jected to aminolysis by the treatment of 50% hydro-
xylamine aqueous solution in the presence of sodium
cyanide in methanol, to afford compounds 12b–d in 51-
72% yield. Finally, 3,4-O-isopropylidene group in 12b–d
were cleaved by treatment of Muromac1 to provide the
target compounds 1b–d in 66–78% yields.14 Compounds
2–5 were synthesized, according to Scheme 2, which was
same manner to Scheme 1. Compounds 16a,b were
subjected to hydrolysis by 1 N sodium hydroxide, fol-
lowed by the condensation of benzyloxyamine hydro-
chloride (NH2OBn) in the presence of 1-(3-
Finally, compounds 17a,b were hydrogenated in the
presence of 10% palladium-carbon under hydrogen
atmosphere to afford target compounds 2,3 in 69–71%
yields.14 On the other hand, after conversion of com-
pound 16c into 16d, compounds 16c,d were subjected to
aminolysis, to afford compounds 18c,d in 49–62%
yields. Finally, 3,4-O-isopropylidene group in 18c,d
were cleaved by treatment of Muromac1 to provide the
target compounds 3,4 in good yields.14
Biological Evaluation
dimethylamino-propyl)-3-ethyl-carbodiimide
hydro-
chloride (WSC) and 1-hydroxy-triazole (HOBt) and the
deprotection of isopropylidene group using Muromac1,
to give compounds 17a,b in moderate yield (Scheme 2).
Inhibitory activities against TACE and MMPs (MMP-
1, MMP-3, MMP-9) of compound 1a–d and 2–5 were
summarized in Table 1.15 At first, we focused on the

H. Moriyama et al. / Bioorg. Med. Chem. Lett. 13 (2003) 2737–2740
2739
Scheme 2. (a) 10% Pd-C/H2, EtOAc; (b) 7a–c, 4-DMAP, DMF, 69–87% from 6; (c) Muromac, 90% MeOH; (d) cerium chloride heptahydrate,
oxalic acid, CH3CN, 49–68%; (e) methanesulfonyl chloride, Et3N, CH2Cl2, 47–61%; (f) K2CO3, DMF, 79–93%; (g) 1N NaOH; (h) NH2OBn, WSC,
HOBt, DMF, 46–51% from 16a,b; (i) Muromac, 90–93%; (j) 10% Pd-C/H2, MeOH, 69–71%; (k) (i) 10% Pd-C/H2, EtOAc; (ii) cyclohexanol,
DEAD, Ph3P, THF, 49%; (l) 50% NH2OH aq, NaCN, MeOH, 49–62%; (m) Muromac, 82–85%.
selective spectrum for inhibitory activities toward
MMPs versus TACE.
Table 1. Inhibitory activities against MMP-1,3,9, and TACE
Compd
rMMP-1
Ki (nM)
rMMP-3
Ki (nM)
rMMP-9
Ki (nM)
TACE
Ki (nM)
Next, we investigated the conversion of arylsulfonyl-
amide (P10-substituent). The stereo-chemistry of azasu-
gar scaffold was fixed as 2R,3R,4R,5S-configuration,
due to the potent inhibitory activities of compound 1b
against MMPs and TACE. Compounds 2 and 4,
1a
1b
1c
1d
2
3
4
5
84
25
>850
450
>850
8.0
850
1.7
7.7
490
85
42
0.51
2.6
157
4.8
780
82
71
12
510
340
8.7
2.3
1.6
67
64
having
2-ethoxyethoxy and benzyloxy moieties,
0.06
6.1
0.93
respectively, exhibited weak activity against MMP-1.
This result indicated that 4-(2-ethoxyethoxy)- and 4-
benzyloxy-benzenesulfonylamide units would not be
accommodated to shallow S10 pocket in MMP-1.17 On
the other hand, compound 3, bearing phenoxy moiety,
showed most potent inhibitory activities against MMP-
1, -3, -9 and TACE, and was 5-80 times more potent
than methoxy type 1b. In addition, inhibitory activities
of compound 3 toward MMP-3 and -9 were 165–183
times potent than those of Marimastat, a well-known
MMP inhibitor. Interestingly, in the case of cyclohex-
yloxy type 5, it was found that inhibitory activities
against MMP-1, -9 and TACE were dramatically
decreased, compared to compound 3. From these results, it
was suggested that the tail end phenoxy moiety such as in
compound 3 would be more preferable for inhibitory
activity against MMPs and TACE than the other sub-
stituents.
100
1.8
Marimastat
1.1
84
11
0.40
stereochemistry of azasugar component. Compound 1b
bearing 2R,3R, 4R,5S-configuration exhibited potent
inhibitory activities against all metalloproteinases
(MMP-1, -3, -9 and TACE), Ki values were 25 nM
against MMP-1, 7.7 nM against MMP-3, 4.8 nM
against MMP-9 and 12 nM against TACE, respectively.
Accordingly, it was clarified that 1b showed 2–33 times
more potent inhibitory activities against MMP-1, -3, -9
and TACE than compound 1a having 2R,3S,4R,5S-
configuration. On the other hand, compound 1c having
2R,3S,4S,5S-configuration was less against all four tar-
get enzymes. Moreover, compound 1d with
2R,3S,4S,5R-configuration was also a weak-moderate
metalloproteinase inhibitor. Although R-configuration
at the 2-position of azasugar compound would be
essential for binding to MMPs and TACE alike other
sulfonamide-based inhibitors,15,16 it was found that ste-
reochemistry at the 3-, 4-, 5-positions as well as the 2-
position of azasugar scaffold would be crucial for the
interaction with MMP-1, -3, -9 and TACE. In addition,
appropriate stereochemistry at the 3-, 4-, 5-positions of
azasugar skeleton could be expected to show broad or
In conclusion, we have synthesized a series of azasugar-
based MMP/ADAM inhibitors to develop compound
1a. As a result, we succeeded to find compound 3,
bearing 2R,3R,4R,5S-configuration and 4-phenoxy-
benzenesulfonylamide moiety, which exhibited desirable
potent inhibitory activities against TACE and MMP-1,
-3, -9. In addition, azasugar could be expected to
improve water solubility, compared to other classes of
metalloproteinase inhibitor. Therefore, azasugar-based

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H. Moriyama et al. / Bioorg. Med. Chem. Lett. 13 (2003) 2737–2740
compound would be a promising candidate of thera-
peutic agent for diseases associated with metalloprotei-
nases. We are now investigating in vivo test of azasugar
compound 3.
9. Morimoto, Y.; Nishikawa, K.; Ohashi, M. Life Sci. 1997,
61, 795.
10. Nelson, F. C.; Zask, A. Exp. Opin. Ther. Pat. 1999, 8, 383.
11. Moriyama, H.; Tsukida, T.; Inoue, Y.; Kondo, H.; Yosh-
ino, K.; Nishimura, S. Bioorg. Med. Chem. Lett. in press.
12. (a) Kim, J. H.; Yang, M. S.; Lee, W. S.; Park, K. H. J. Chem.
Soc., Perkin Trans. 1 1998, 2877. (b) Lee, B. W.; Jeong, I.-Y.;
Yang, M. S.; Choi, S. U.; Park, K. H. Synthesis 2000, 1305.
13. (a) Park, K. H.; Yoon, Y. J.; Lee, S. G. Tetrahedron Lett.
1994, 35, 9737. (b) Xiao, X.; Bai, D. Synlett 2001, 535.
14. All new compounds gave satisfactory characteristics data.
Characteristics are given for a representative compound: 1b:
1H NMR (DMSO-d6, 250 MHz) d 3.0–3.7 (m, 5H), 3.84 (s,
3H), 4.20 (d, 1H, J=5.3 Hz), 7.09 (d, 2H, J=9.0 Hz), 7.67 (d,
2H, J=9.0 Hz), 8.82 (s, 1H), 10.67 (s, 1H). MALDI-TOF: 385
(M+Na+), 401 (M+K+).
Acknowledgements
The present work was supported by a grant for
‘Research and Development on Glycocluster Control-
ling Biomolecules’ from the New Energy and Industrial
Technology Development Organization (NEDO).
15. (a) Recombinant human collagenase-1 (MMP-1), strome-
lysin-1 (MMP-3), gelatinase B (MMP-9) and TNF-a convert-
ing enzyme (TACE) were used in our studies. Assay conditions
were referred as below: Sawa, M.; Kiyoi, T.; Kurokawa, K.;
Kumihara, H.; Yamamoto, M.; Miyasaka, T.; Ito, Y.; Hir-
ayama, R.; Inoue, T.; Kirii, Y.; Nishiwaki, E.; Ohmoto, H.;
Maeda, Y.; Ishibushi, E.; Inoue, Y.; Yoshino, K.; Kondo, H.
J. Med. Chem. 2002, 45, 919. (b) Yoshiizumi, K.; Yamamoto,
M.; Miyasaka, T.; Ito, Y.; Kumihara, H.; Sawa, M.; Kiyoi, T.;
Yamamoto, T.; Nakajima, F.; Hirayama, R.; Kondo, H.; Ish-
ibushi, E.; Ohmoto, H.; Inoue, Y.; Yoshino, K. Bioorg. Med.
Chem. 2003, 11, 433.
16. Tamura, Y.; Watanabe, F.; Nakatani, T.; Yasui, K.; Fuji,
M.; Komurasaki, T.; Tsuzuki, H.; Maekawa, R.; Yoshioka,
T.; Kawada, K.; Sugita, K.; Ohtani, M. J. Med. Chem. 1998,
41, 640.
17. (a) Yamamoto, M.; Tsujishita, H.; Hori, N.; Ohishi, Y.;
Inoue, S.; Ikeda, S.; Okada, Y. J. Med. Chem. 1998, 41, 1209.
(b) Lovejoy, B.; Welch, A. R.; Carr, S.; Luong, C.; Broka, C.;
Hendricks, R. T.; Campell, J. A.; Walker, K. A. M.; Martin,
R.; Van Wart, H.; Browner, M. F. Nat. Struct. Biol. 1999, 6,
217.
References and Notes
1. Michaelides, M. R.; Curtin, M. L. Curr. Pharm. Des. 1999,
5, 787.
2. Whittaker, M.; Floyd, C. D.; Brown, P.; Gearing, A. J. H.
Chem. Rev. 1999, 2735.
3. Beckett, R. P.; Whittaker, M. Exp. Opin. Ther. Pat. 1998,
8, 259.
4. Rothenberg, M. L.; Nelson, A. R.; Hande, K. R. Oncolo-
gist 1998, 3, 271.
5. Black, R. A.; White, J. M. Curr. Opin. Cell. Biol. 1998, 10,
654.
6. Bemelmans, M. H.; van Tits, L. J.; Buurman, W. Crit. Rev.
Immunol. 1996, 16, 1.
7. Moreland, L. W.; Baumgartner, S. W.; Schiff, M. H.; Tin-
dall, E. A.; Fleischmann, R. M.; Weaver, A. L.; Ettlinger,
R. E.; Cohen, S.; Koopman, W. J.; Mohler, K. N. Engl. J.
Med. 1997, 337, 141.
8. Clements, J. M.; Cossins, J. A.; Wells, G. M.; Corkill, D. J.;
Helfrich, K.; Wood, L. M.; Pigott, R.; Stabler, G.; Ward, G. A.;
Gearing, A. J.; Miller, K. M. J. Neuroimmunol. 1997, 74, 85.