The First L-Iduronic Acid-Type 1-N-Iminosugars Having Inhibitory Activity of Experimental Metastasis

Full text of DOI:10.1021/ja953989b

J. Am. Chem. Soc. 1996, 118, 3051-3052
The First L-Iduronic Acid-Type 1-N-Iminosugars
3051
Having Inhibitory Activity of Experimental
Metastasis
Yoshio Nishimura,* Takahiko Satoh, Hayamitsu Adachi,
Shinichi Kondo, Tomio Takeuchi, Masayuki Azetaka,†
Harumi Fukuyasu,† and Yumiko Iizuka†
Institute of Microbial Chemistry, 3-14-23 Kamiosaki
Shinagawa-ku, Tokyo 141, Japan
Pharmaceutical Research Center, Meiji Seika Kaisha, Ltd.
760 Morooka-cho, Kohoku-ku, Yokohama 222, Japan
ReceiVed NoVember 28, 1995
Tumor cell adhesion to various basement membrane com-
ponents and degradation of extracellular matrix and basement
membranes are an important step of tumor metastasis. â-D-
Glucuronidase and R-L-iduronidase are known to degrade the
mammalian glycosaminoglycans, the major constituents of
endothelial basement membranes.1,2 Heparanase (end-â-glu-
curonidase) activity was also proved to correlate with lung
colonization abilities of murine B16 melanoma cells by extra-
cellular matrix degradation and to be inhibited by heparanase
inhibitors.3
Figure 1.
Scheme 1
Siastatin B (1) was isolated as an inhibitor of neuraminidase
and â-glucuronidase from Streptomyces culture,4 and the
6-epimer of 1 has recently been shown to be an inhibitor for
heparanase.5 This discovery stimulated interests in the synthesis
of specific glucuronidase inhibitors for antimetastasis of tumor
cells and led to highly potent â-glucuronidase inhibitors 2 and
3. They showed the inhibition of invasion of B16 variant (B16
BL6) and Lewis lung carcinoma (3LL) cells through reconsti-
tuted basement membrane and the significant suppression of
experimental and spontaneous pulmonary metastasis of B16 BL6
and/or 3LL cells in mice.6 Compounds 2 and 3, which
structurally resemble glucuronic acid (4) as a 1-N-iminosugar,7
probably mimic 4 in binding to â-glucuronidase and strongly
inhibit the enzymatic reaction. We speculated from these facts
and results that if the metabolism of R-L-iduronide as well as
† Pharmaceutical Research Center.
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â-D-glucuronide of basement membranes was responsible for
tumor metastasis, L-iduronic acid-type 1-N-iminosugars should
inhibit tumor metastasis. Here, we report the first L-iduronic
acid-type 1-N-iminosugars 6, 7, and 8 having experimental
antimetastatic activity in mice.
In order to synthesize 1-N-iminosugar corresponding to
L-sugar, the configurational inversion of the carboxyl group of
1 was examined. After unsuccessful attempts of epimerization
of the carboxyl group of some ester derivatives of 1 with bases
and a nonstereoselective epimerization by a conjugated Michael
addition of the alcohol to the R,â-unsaturated ester 10 with base,8
attention was directed to the intramolecular Michael addition
of O-imidate9 to R,â-unsaturated ester 11. Compound 11
underwent smoothly cis oxyamination to give the oxazoline 13
in 76% yield and a trace amount of its epimer 14. The
intermediate imidate anion 12 from reaction with CCl3CN
underwent efficient conjugate addition without the use of an
electrophile to trigger oxazoline formation. Hydrolysis of 13
(p-CH3C6H4SO3H, C6H5N/H2O)10 gave 15 and 16 in yields of
77 and 9%, respectively. Reductive cleavage of the trichloro-
(7) (a) Nishimura, Y. In StereoselectiVe synthesis and transformation of
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Viral infection and tumor metastasis; Atta-ur-Rahman, Ed.; Studies in
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Lundt, I.; Bols, M. Angew. Chem., Int. Ed. Engl. 1994, 33, 1778.
(8) (a) Nishimura, Y.; Kudo, T.; Umezawa, Y.; Kondo, S.; Takeuchi, T.
Nat. Prod. Lett. 1992, 1, 39. (b) Nishimura, Y.; Umezawa, Y.; Kondo, S.;
Takeuchi, T.; Mori, K.; Kijima-Suda, I.; Tomita, K.; Sugawara, K.;
Nakamura, K. J. Antibiot. 1993, 46, 1883.
(9) Alonso, R. A.; Burgey, C. S.; Rao, B. V.; Vite, G. D.; Vollerthun,
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0002-7863/96/1518-3051$12.00/0 © 1996 American Chemical Society

3052 J. Am. Chem. Soc., Vol. 118, No. 12, 1996
Communications to the Editor
acetyl group with NaBH411 then gave 17 and 18 in good yields.
Thus obtained 17 and 18 were smoothly converted into 6 and
7 by treatment with acid, respectively. The guanidine moiety
is an important feature in many biological active compounds,
especially in binding to the enzyme such as influenza viral
N-acetylneuraminidase.12,13 The major isomer 17 was then
converted to the bis-Boc-protected guanidine 19 ((BocNH)2CS,
HgCl2)14 in excellent yield. Compound 19 was straightfor-
tumor cells.17 Since the inhibitory activity of siastatin B
analogues for experimental pulmonary metastasis is well
proportional to that for spontaneous pulmonary metastasis,6c the
experimental metastasis assay was employed for this study. As
expected, pulmonary colonization after intravenous transplanta-
tion of B16 BL6 cells into the tail veins of mice was significantly
suppressed dose-dependently by in Vitro pretreatment with 6,
7, and 8 (44 and 31% inhibition at 172 µM of 6 and 7 (P <
0.05), respectively; 91 and 97% inhibition at 90 and 150 µM
of 8 (P < 0.001), respectively).15 Compounds 6, 7, and 8 had
no significant effects on cell growth at the concentrations used
in this study (data not shown). Compound 8 was also inhibitory
against B16 BL6 cell invasion through reconstituted basement
membranes (59% inhibition at 870 µM (P < 0.01)).15 These
facts indicate that the antimetastatic effect of 8 is due to its
anti-invasive rather than antiproliferative activity. On the other
hand, molecular modeling15 using PM3 in MOPAC revealed
that 8 superimposes well on L-iduronic acid (5) and has the
hydroxyl and carboxyl groups lying in the same region of space
as those of 5, with the acetamide and guanidino moiety also
being topographically equivalent to the hydroxyl moieties of 5.
It is highly likely that in contrast with â-glucuronidase inhibitors
2 and 3, compound 8 mimics L-iduronic acid in binding to
L-iduronidase and inhibits the enzymatic hydrolysis. These
findings suggest that the metabolism of R-L-iduronide of the
basement membranes of a normal cell and/or tumor cell may
participate in human melanoma metastasis and that melanoma
cells may secrete not only heparanase but also R-L-iduronidase.
The present study shows that the L-iduronic acid-type 1-N-
iminosugars are a promising candidate of new drugs for tumor
metastasis.
6
wardly transformed into 8 by treatment with acid. The C3-
conformations as well as stereochemistries of 6, 7, and 8 were
1
established by H NMR spectra.15
As expected, all analogues did not inhibit D-glycosidases16
(IC50 > 0.39 mM). These results indicate that the analogues
having 6C3-conformation are significantly distinct from the
hitherto known analogues7 of 1 having 3C6-conformation on the
specificity against D-sugar hydrolases. These analogues were
then assayed for anti-invasive and antimetastatic activities of
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(15) See supporting information.
(16) Inhibitory activities against Baker’s yeast R-glucosidase,16a almond
â-glucosidase,16b Jack beans R-mannosidase,16c snail â-mannosidase,16c
Escherichia coli R- and â-galactosidase,16d bovine liver â-glucuronidase,16e
chicken liver R-N-acetylgalactosaminidase,16f and bovine epididymis â-N-
acetylglucosaminidase16g were assayed by similar methods described in
references. All enzymes were purchased from Sigma Chemical Company.
(a) Halvorson, H. O.; Fllias, L. C. Biochim. Biophys. Acta 1958, 30, 28.
(b) Kobayashi, A. Agr. Biol. Chem. 1962, 26, 303. (c) Li, Y.-T. J. Biol.
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J. Biol. Chem. 1965, 240, 2468. (e) Stahl, P. P. D.; Fishman, W. H. In
â-D-Glucuronidase. â-D-Glucuronide glucuronosohydrolase, EC 3.2.1.31;
Voight, K. D., Ed.; Methods of Enzymatic Analysis; Academic Press: New
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Supporting Information Available: Experimental procedures and
characterization data for new compounds, biological data for 6, 7, and
8, and PM3/MOPAC optimized structures of 8 and R-L-iduronic acid
(9 pages). This material is contained in many libraries on microfiche,
immediately follows this article in the microfilm version of the journal,
can be ordered from the ACS, and can be downloaded from the Internet;
see any current masthead page for ordering information and Internet
access instructions.
JA953989B
(17) R-L-Iduronidase is now not available for us.
(18) Kudo, T.; Nishimura, Y.; Kondo, S.; Takeuchi, T. J. Antibiot. 1993,
46, 300.