ORGANIC

LETTERS

Expedient Synthesis of Triazole-Linked 2004

Vol. 6, No. 18
Glycosyl Amino Acids and Peptides 3123-3126

Brian H. M. Kuijpers,† Stan Groothuys,† A. (Bram) R. Keereweer,†

Peter J. L. M. Quaedflieg,‡ Richard H. Blaauw,§ Floris L. van Delft,† and
Floris P. J. T. Rutjes*,†

Department of Organic Chemistry, NSRIM, UniVersity of Nijmegen, ToernooiVeld 1,

6525 ED Nijmegen, The Netherlands, DSM Research, Life SciencessAdVanced
Synthesis, Catalysis and DeVelopment, P.O. Box 18, 6160 MD Geleen, The
Netherlands, and Chiralix B.V., ToernooiVeld 100, 6525 EC Nijmegen, The Netherlands
rutjes@[Link]

Received June 18, 2004

ABSTRACT

An expedient, high-yielding synthesis of two types of triazole-linked glycopeptides is described. These novel and stable glycopeptide mimics
were prepared via Cu(I)-catalyzed [3 + 2] cycloaddition of either azide-functionalized glycosides and acetylenic amino acids or acetylenic
glycosides and azide-containing amino acids.

Glycopeptides1 constitute a class of natural compounds, these syntheses, however, are rather complex and often
involved in a number of important biological functions. By feature low overall yields.4 Our research efforts aim at an
far the most commonly encountered members of this family efficient, high-yielding synthesis of triazole-linked glyco-
are N- and O-linked glycopeptides.2 Synthesis of such peptides such as 1 and 2 (Scheme 1), via a mild, Cu-catalyzed
glycopeptides is complicated by the sensitivity of the
glycosidic linkage between the (oligo)saccharide and the
peptide toward chemical and enzymatic hydrolysis. Synthesis Scheme 1. Retrosynthesis
of (unnatural) amino acids, with the amino acid side chain
connected to the sugar unit via an isosteric linkage, may lead
to chemically and metabolically more stable analogues with
potential biological activity3 (e.g., inhibitory activity toward
glycosidases) or provide means to elucidate biochemical
pathways.
Approaches to obtain stable glycopeptide analogues often
involve the synthesis of C-linked glycopeptides. Most of
† University of Nijmegen.
‡ DSM Research.
§ Chiralix B.V.
(1) Glycopeptides and Related Compounds: Synthesis, Analysis and
Applications; Large, D. G., Warren, C. D., Eds.; Marcel Dekker: New York, procedure for the [3 + 2] cycloaddition between organic
1997.
(2) (a) Kunz, H. Angew. Chem., Int. Ed. 1987, 26, 294. (b) Spiro, R. G. azides and acetylenes (“click reactions”). As recently inde-
Glycobiology 2002, 12, 43R. (c) Seitz, O. ChemBioChem 2000, 1, 214. pendently reported by the groups of Meldal5 and Sharpless,6
(3) (a)Wu, T. C.; Goekjian, P. G.; Kishi, Y. J. Org. Chem. 1987, 52,
4819. (b) Haneda, T.; Goekjian, P. G.; Kim, S. H.; Kishi, Y. J. Org. Chem. this reaction generally results in the corresponding 1,4-
1992, 57, 490. disubstituted 1,2,3-triazoles in high yields. Application of
10.1021/ol048841o CCC: $27.50 © 2004 American Chemical Society
Published on Web 07/31/2004
this reaction to either azidoglycosides 3 and acetylenic amino
acids 4 or acetylenic glycosides 5 and azide-containing amino Table 1. Variation of the Carbohydrate Moiety
acids 6 would then result in the triazole-linked glycopeptides
1 and 2, respectively, which constitute a novel compound
class.7 Besides the aim of constructing stable glycopeptide
mimics, the resulting substituted triazoles may display
relevant biological activity against various targets.8
To probe the viability of our approach, various glycosides
containing an anomeric azide functionality were prepared
and reacted with (R)-N-Boc-propargylglycine methyl ester
(7, Table 1).9 The coupling was initially studied using
azidoglucoside (8) and azidogalactoside (9)10 in combination
with different Cu(I) species (e.g., CuI, CuCl, and CuCN)
and different bases (e.g., Et3N and DIPEA). Optimal results
were obtained using modified Sharpless conditions,6 involv-
ing 0.2 equiv of Cu(OAc)2 and 0.4 equiv of sodium ascorbate
in a 1:1 (v/v) mixture of H2O and tert-BuOH. This eventually
provided the targeted glycoamino acids 15 and 16 in 98 and
88% yields (entries 1 and 2).
Under the same conditions, the benzylated glycosyl azide
1011 reacted smoothly to give the corresponding adduct 17
with retention of the anomeric configuration (entry 3).
Likewise, the glucosamine- and galactosamine-derived
azides 11 and 12,12 respectively, reacted in an efficient
manner with the acetylenic amino acid 7 to give the

(4) For entries into C-glyco amino acid synthesis, see for example: (a)
Taylor, C. M. Tetrahedron 1998, 54, 11317. (b) Du, Y.; Lindhardt, R. J.
Tetrahedron 1998, 54, 9913. (c) Dondoni, A.; Marra, A. Chem. ReV. 2000,
100, 4395. (d) Vincent, S. P.; Schleyer, A.; Wong, C.-H. J. Org. Chem.
2000, 65, 4440. (e) Xu, X.; Fakha, G.; Sinou, D. Tetrahedron 2002, 58,
7539. (f) Lane, J. W.; Halcomb, R. L. J. Org. Chem. 2003, 68, 1348 (g)
Turner, J. J.; Leeuwenburgh, M. A.; Van der Marel, G. A.; Van Boom, J.
H. Tetrahedron 2001, 42, 8713. (h) Dondoni, A.; Mariotti, G.; Marra, A.;
Massi, A. Synthesis 2001, 14, 2129. (i) Westermann, B.; Walter, A.; Flörke,
U.; Altenbach, H.-J. Org. Lett. 2001, 3, 1375. (j) McGarvey, G. J.; Benedum,
T. E.; Schmidtmann, F. W. Org. Lett. 2002, 4, 3591. (k) Dondoni, A.;
Giovannini, P. P.; Marra, A. J. Chem. Soc., Perkin Trans. 1 2002, 2380. (l)
Nolen, E. G.; Kurish, A. J.; Wong, K. A.; Orlando, M. D. Tetrahedron
Lett. 2003, 44, 2449. (m) Palomo, C.; Oiarbide, M.; Landa, A.; Concepción
González-Rego, M.; Garcı́a, J. M.; Gonzáles, A.; Odriozola, J. M.; Martı́n-
Pastor, M.; Linden, A. J. Am. Chem. Soc. 2002, 124, 8637.
(5) Tornøe, C. W.; Christensen, C.; Meldal, M. J. Org. Chem. 2002, 67,
3057.
(6) Rostovtsev, V. V.; Green, L. G.; Fokin, V. V.; Sharpless, K. B.
Angew. Chem. Int. Ed. 2002, 41, 2596.
(7) For earlier reports on [3 + 2] cycloadditions of azidoglycosides and
acetylenes, see: (a) Bröder, W.; Kunz, H. Carbohydr. Res. 1993, 249, 221.
(b) Al-Masoudi, N. A.; Al-Soud, Y. A. Tetrahedron Lett. 2002, 43, 4021. a Reagents and conditions: 1 equiv of azidoglycoside, 1 equiv of amino
(c) Marco-Contelles, J.; Jiménez, C. A. Tetrahedron 1999, 55, 10511 (d)
Peto, C.; Batta, G.; Gyorgydeak, Z.; Sztaricskai, F. Carbohydr. Chem. 1996, acid derivative, 0.2 equiv of Cu(OAc)2, 0.4 equiv of sodium ascorbate, H2O/
15, 465 and references therein tert-BuOH 1:1 (v/v), rt, 16 h. b Yield of isolated product.
(8) For examples of biologically active triazoles, see: (a) Alvarez, R.;
Velazquez, S.; San, F.; Aquaro, S.; De, C.; Perno, C. F.; Karlsson, A.; corresponding products 18 and 19 in good yields (entries 4
Balzarini, J.; Camarasa, M. J. J. Med. Chem. 1994, 37, 4185. (b) Velazquez,
S.; Alvarez, R.; Perez, C.; Gago, F.; De, C.; Balzarini, J.; Camarasa, M. J. and 5). The benzoyl-protected glucosamine 1313 surprisingly
AntiVir. Chem. Chemother. 1998, 9, 481. (c) Genin, M. J.; Allwine, D. A.; gave a somewhat lower yield of the glycosylamino acid 20
Anderson, D. J.; Barbachyn, M. R.; Emmert, D. E.; Garmon, S. A.; Graber,
D. R.; Grega, K. C.; Hester, J. B.; Hutchinson, D. K.; Morris, J.; Reischer, (entry 6). Finally, the 2-azido functionalized glucose deriva-
R. J.; Ford, C. W.; Zurenko, G. E.; Hamel, J. C.; Schaadt, R. D.; Stapert, tive 1414 gave the desired coupling product in 77% yield
D.; Yagi, B. H. J. Med. Chem. 2000, 43, 953.
(9) Enantiopure propargylglycine is commercially available but in our
(entry 7). Clearly, there were no notable differences in
hands was prepared via an enzymatic resolution process: (a) Wolf, L. B.;
Sonke, T.; Tjen, K. C. M. F.; Kaptein, B.; Broxterman, Q. B.; Schoemaker, (12) Prepared via tetra-O-acetyl-R-D-pyranosyl chloride from the corre-
H. E.; Rutjes, F. P. J. T. AdV. Synth. Catal. 2001, 343, 662. (b) Sonke, T.; sponding glucosamine and galactosamine (ref 13) and subsequent treatment
Kaptein, B.; Boesten, W. H. J.; Broxterman, Q. B.; Kamphuis, J.; Formaggio, with NaN3: Lehnhoff, S.; Goebel, M.; Karl, R. M.; Kloesel, R.; Ugi, I.
F.; Toniolo, C.; Rutjes, F. P. J. T.; Schoemaker, H. E. In StereoselectiVe Angew. Chem., Int. Ed. 1995, 34, 1104.
Biocatalysis; Patel, R. N., Ed.; Marcel Dekker: New York, 2000; p 23. (13) Prepared by reaction of the benzoylated thioglycoside with ICl and
(10) Shiozaki, M.; Arai, M.; Macindoe, W. M.; Mochizuki, T.; Kurakata, Me3SiN3, respectively.
S.; Maeda, H.; Nishijima, M. Chem. Lett. 1996, 9, 735 (14) Prepared from the corresponding glucosamine via azido transfer,
(11) Prepared from 2,3,4,6-tetra-O-benzylglucose by treatment with DBU followed by acetylation: Alper, P. B.; Hung, S.; Wong, C.-H. Tetrahedron
and DPPA: Mizuno, M.; Shioiri, T. Chem. Commun. 1997, 2165. Lett. 1996, 37, 6029.

3124 Org. Lett., Vol. 6, No. 18, 2004

reactivity between the several monosaccharides or between
the R- and â-isomers. Preliminary experiments to determine Table 3. Cycloaddition Reactions Involving a Dipeptide
the stability of the triazole linkage in compound 15 revealed
that this moiety is stable under a variety of acidic and basic
conditions.15
Table 2 shows the application of different acetylenic amino
acids containing a variety of amino protecting groups in the

Table 2. Variation of the Amino Acid Moiety

a Reagents and conditions: 1 equiv of azidoglycoside 8, 1 equiv of

dipeptide, 0.2 equiv of Cu(OAc)2, 0.4 equiv of sodium ascorbate, H2O/
tert-BuOH 1:1 (v/v), rt, 16 h. b Yield of isolated product.

products 30 and 31 in good yields. Similarly, R-methyl-

substituted propargylglycine 26 (entry 5) afforded the gly-
coamino acid 32 in 85% yield.
In addition, subjection of the acetylenic aminonitrile 2717
gave the desired adduct 33 in 60% yield.
Tables 3 and 4 show that the scope of the click reactions
can be extended to dipeptides and disaccharides, respectively.

Table 4. Cycloaddition Reactions Involving a Disaccharide

a Reagents and conditions: 1 equiv of azidoglycoside, 1 equiv of amino

acid derivative, 0.2 equiv of Cu(OAc)2, 0.4 equiv of sodium ascorbate, H2O/
tert-BuOH 1:1 (v/v), rt, 16 h. b Yield of isolated product.

a Reagents and conditions: 1 equiv of azidoglycoside 38, 1 equiv of

amino acid, 0.2 equiv of Cu(OAc)2, 0.4 equiv of sodium ascorbate, H2O/
cycloaddition with azidoglycoside 8. From Table 1, the tert-BuOH 1:1 (v/v), rt, 16 h. b Yield of isolated product.
compatibility of the Boc group with these reaction conditions
was already clear; entries 1 and 2 of Table 2 now show that
Fmoc and Ts are also well-suited for this reaction. Increasing For example, the two protected dipeptides 34 and 3518 were
the length of the side chain of the amino acid16 (entries 3 successfully coupled to glycosyl azide 8, affording the
and 4) showed no significant change in reactivity, providing glycopeptides 36 and 37 in 51 and 92% yields, respectively.

(15) Triazole linkage appeared to be stable using, for example: 2.5 M (17) Prepared from Boc-protected (S)-propargylglycine amide via de-
HCl in EtOAc, 2 h, rt; 1 M aqueous HCl, >5 days, rt; 1 M HCl in MeOH, hydration: Cossu, S.; Giacomelli, G.; Conti, S.; Falorni, M. Tetrahedron
6 h, reflux; K2CO3 in MeOH, 4 h, rt; 1.25 M aqueous NaOH, 24 h, reflux. 1994, 50, 5083.
(16) van Esseveldt, B. C. J.; van Delft, F. L.; de Gelder, R.; Rutjes, F. (18) Dipeptides 34 and 35 were synthesized using a standard coupling
P. J. T. Org. Lett. 2003, 5, 1717. reaction involving PyBOP.

Org. Lett., Vol. 6, No. 18, 2004 3125

Inversely, the disaccharide azide 3819 was coupled to amino
acid 7 and dipeptide 34 in satisfactory yields (Table 4). Scheme 2. Cycloaddition between an R-Acetylenic Glycoside
Considering the fact that disaccharides and dipeptides couple and Azide-Containing Amino Acids
very well under these conditions, we anticipate that also
larger and more complex oligosaccharides and oligopeptides
can be successfully ligated in this way.
Since all examples so far comprise the combination of
azidoglycosides with acetylenic amino acids, we were
intrigued whether ligation of constituents with inverted
functionality, with the triazole-N1 position bound to the
amino acid, would also be feasible. To this end, acetylenic
glycosides and azide-containing amino acid derivatives were a Reagents and conditions: 1 equiv of acetylenic glycoside 46,
prepared: the R- and â-acetylenic glucose derivatives 41 and 1 equiv of azide, 0.2 equiv of Cu(OAc)2, 0.4 equiv of sodium
46, respectively, from 2,3,4,6-tetra-O-benzyl-D-glucose,20 and ascorbate, H2O/tert-BuOH 1:1 (v/v), rt, 16 h. bYield of isolated
the required amino acid azides (42a-c and 43) from the product.
corresponding amines via azido transfer using TfN3.21
Evidently, these “reversed” triazole-linked glycopeptides may explain the slightly lower yields. Furthermore, there is
were also readily obtained via the Cu-catalyzed [3 + 2] again virtually no change in reactivity upon elongation of
cycloaddition. Both the R- and â-glycopeptide products were the amino acid side chain as can be inferred from Table 5,
formed uneventfully in yields ranging from 60 to 84% (Table entries 1-3, and Scheme 2.
5 and Scheme 2). In general, amino acid derivatives In conclusion, we have developed a straightforward,
versatile, and high-yielding method for the synthesis of a
novel class of glycopeptides. Both R- and â-triazole-linked
Table 5. Cycloaddition between a â-Acetylenic Glycoside and glycopeptides can be efficiently prepared using a variety of
Azide-Containing Amino Acids suitably functionalized (oligo)saccharides and (oligo)peptides.
Currently, we aim at further expanding the scope of this
application and more specifically at the synthesis of triazole-
containing mimics of biologically active glycopeptides.

Acknowledgment. S. A. M. W. van den Broek, B. C. J.

van Esseveldt, B. Verheijen, and M. IJsselstijn are kindly
entry amino acid n R glycopeptidea yield (%)b acknowledged for the preparation of several compounds.
Senter (Ministry of Economic Affairs, The Netherlands) is
1 42a 1 H 44a 70%
2 42b 2 h 44b 73% gratefully acknowledged for providing financial support.
3 42c 3 H 44c 71%
4 43 1 Me 45 84% Supporting Information Available: Experimental pro-
a Reagents and conditions: 1 equiv of acetylenic glycoside 41, 1 equiv
cedures and spectroscopic data of the triazole-linked glyco-
of azide, 0.2 equiv of Cu(OAc)2, 0.4 equiv of sodium ascorbate, H2O/tert- peptides. This material is available free of charge via the
BuOH 1:1 (v/v), rt, 16 h. b Yield of isolated product. Internet at [Link]
OL048841O
containing a free carboxylic acid appeared to be somewhat
(20) Dondoni, A.; Mariotti, G.; Marra, A. J. Org. Chem. 2002, 67, 4475.
harder to purify than their methyl ester counterparts, which (21) (a) Rosenberg, S. H.; Spina, K. P.; Woods, K. W.; Polakowski, J.;
Martin, D. L.; Yao, Z. L.; Stein, H. H.; Cohen, J.; Barlow, J. L.; Egan, D.
(19) Hepta-O-acetyl-â-lactosyl azide (38) was prepared from lactose by A.; Tricarico, K. A.; Baker, W. R.; Kleinert, H. D. J. Med. Chem. 1993,
peracetylation followed by treatment with SnCl4 and Me3SiN3.8 36, 449. (b) Lundquist, J. T.; Pelletier, J. C. Org. Lett. 2001, 3, 781.

3126 Org. Lett., Vol. 6, No. 18, 2004