Click multivalent heterogeneous neoglycoconjugates - modular synthesis and evaluation of their binding affinities

FULL PAPER

DOI: 10.1002/ejoc.200801169

Click Multivalent Heterogeneous Neoglycoconjugates – Modular Synthesis and

Evaluation of Their Binding Affinities

Mariano Ortega-Muñoz,^[a]Francisco Perez-Balderas,^[a]Julia Morales-Sanfrutos,^[a]

Fernando Hernandez-Mateo,^[a]Joaquín Isac-García,^[a]and Francisco Santoyo-Gonzalez*^[a]

Dedicated to Professor Hans Helmut Baer

Keywords: Carbohydrates / Glycoconjugates / Multivalency / Cycloaddition / Click chemistry / Ligand design

The efficient synthesis of structurally diverse, multivalent,

heterogeneous neoglycoconjugates by means of a series of

constitutive sugars and length of the linker. The influence of

those structural parameters on the binding affinities of these

different click-based strategies is described. The methodol- glycomimics toward Concanavalin A (Con A) was evaluated.

ogy is highly efficient and versatile allowing for easy access

to a series of mannose (α-Man)-containing glycoconjugates

differing in their valency, nature of the scaffold, nature of the

Germany, 2009)

recognition process. In spite of this, a limited number of

synthetic, multivalent, heterogeneous neoglycoconjugates

containing different sugar epitopes has been described to

date.^[10–18]

Introduction

Carbohydrates are implicated in a number of important

biological processes involving highly specific events in cell-

cell recognition, cell-protein interactions, and the targeting

of hormones, antibodies, and toxins.^[1]In these processes,

multivalent interactions^[2]are especially prevalent,^[3]where

high-avidity interactions take place between multimeric,

membrane-bound, carbohydrate-binding proteins (lectins)

and their associated carbohydrate ligands, despite the low

affinity provided by a typical monovalent interaction. A

large variety of neoglycoconjugates^[4]ranging from small

molecules to macro- and supramolecular entities have been

synthesized by chemists in attempts to understand, mimic,

and perturb the natural, multivalent, lectin-carbohydrate in-

teractions. By using these glycomimetics, it has been eluci-

dated that factors^[5]such as the rigidity, spacing, topology,

and density of saccharides influence the avidity of multi-

valent carbohydrate-displaying structures for their respec-

tive lectins. The principal architecture of neoglycoconju-

gates usually comprise a core molecule of diverse type that

serve as an oligovalent scaffold for the grafting sugar mole-

cules of the same nature giving rise to chemically well-de-

fined, homogeneous neoglycoconjugates such as glycoclus-

ters, glycodendrimers,^[6]glycocyclodextrins,^[7,8]glycocalixar-

enes,^[8]and glycopolymers.^[9]However, in nature, a number

of different sugar ligands may be essential for a biological

The reported hetero carbohydrate displays having a gly-

cocluster,^[10,11]glycodendrimer,^[12–14]glycocyclodextrin,^[15]

or glycopolymer^[16,17]architecture have been constructed by

means of two different synthetic strategies for the incorpo-

ration of the different saccharide ligands: 1) by using an

orthogonal protection of a polyfunctional core molecule for

the successive attachment of the different sugar moie-

ties^[10,11,13]or 2) by controlling the relative proportions of

the carbohydrate reagents in the reaction with a homogen-

eous, functionalized, reactive core.^[12,14–17]In this last op-

tion, the heterogeneous functionalization can be performed

following a sequential^[12,14,15]or a simultaneous^[16,17]conju-

gation pattern using pure carbohydrate reagents or mixtures

of them, respectively, allowing in this way a modulation of

the sugar density. The covalent attachment of sugar moie-

ties to the selected scaffolds have been performed by means

of amide,^[10,11]thiourea,^[10,12,14]thioether,^[15]and tri-

azole^[13,16,17]linkage constructed by the reaction of sugar

derivatives containing amino, acid, thiocyanate, thiol, and

azide groups, respectively.

The biological studies performed with the heterogeneous,

multivalent, carbohydrate displays have provided some in-

sights into the biological processes in which they are in-

volved. Thus, it has been demonstrated that the sugar den-

sity influences both the clustering rate and the stoichiome-

try of the neoglyconjugate-protein conjugates in assays per-

formed with hetero glycopolymers.^[16,17]In addition, the

[a] Departamento de Química Orgánica, Facultad de Ciencias,

Instituto de Biotecnología,

Universidad de Granada, 18071 Granada, Spain

Fax: +34-958-243-186

E-mail: fsantoyo@ugr.es

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Click Multivalent Heterogeneous Neoglycoconjugates

existence of strong synergistic effects (heterocluster effect)^[15]

on carbohydrate-protein recognition events have been evi-

denced by the comparison of the lectin-binding properties

of highly dense, β-cyclodextrin-centered, homo and hetero

glycoclusters with defined architecture. In a different ap-

proach, supramolecular chemistry principles have been

used^[18]for the generation and deconvolution of dynamic

combinatorial libraries of di- and trivalent hetero glycoclus-

ters.

In this context, the Cu^I-catalyzed azide–alkyne cycload-

dition (CuAAC),^[19]nowadays the best reaction under the

“click chemistry” concept,^[20]appears as an appealing syn-

thetic tool for the construction of complex molecular archi-

tectures considering that its efficiency has been validated by

numerous applications in almost all areas of chemistry

(drug discovery,^[21]bioconjugation,^[22]polymer and material

science,^[23]supramolecular chemistry,^[24]and other related

areas^[25]). In continuing our efforts in the implementation

of the CuAAC methodology for the construction of multi-

valent structures^[26,27]and materials^[28]with a well-defined

architecture, we report herein its applicability for the syn-

thesis of heterogeneous neoglycoconjugates by using a mod-

ular strategy for the successive incorporation of clickable

alkyne and azide sugar derivatives into suitable, comple-

mentary, functionalized scaffolds, thereby extending the re-

sults reported in the preceding contribution of this issue.^[29]

Figure 1. Clickable alkyne and azido sugar derivatives.

We initially prepared divalent neoglycoconjugates by

means of two different strategies. In the first one (Scheme 1),

the click reaction of 1 with the complementary function-

alized Glc and GlcNAc azide derivatives 4 and 5 afforded

the divalent 8 and 9 neoglycoconjugates, respectively, in

which the 1,2,3-triazole ring acts as the bridging unit. We

also prepared the α-Man-homogeneous divalent analogues

10 and 11 by the reaction of 1 with the α-Man-azido deriva-

tives 6 and 7, respectively, to be used as reference com-

pounds in the binding assays.

Results and Discussion

Chemistry

We planned the synthesis of mannose (α-Man)-contain-

ing multivalent heterogeneous neoglycoconjugates differing

in structural parameters such as the valency, length of the

connecting linkers, nature of the scaffolds, and nature of the

constitutive sugars. We pursued two goals: 1) to demon-

strate the applicability of the click-chemistry methodology

as a valuable synthetic tool for accessing complex neoglyco-

conjugates and 2) to evaluate the effect of the structural

parameters of these glycomimics on the receptor binding

affinity for Concanavalin A (Con A) from Canavalia ensi-

formis. We selected this tetrameric plant lectin as a suitable

and widely used model lectin.

We chose the α-Man alkyne (1)^[30]and azido (6 and 7)^[31]

derivatives as easily accessible, clickable, sugar derivatives to

be used in a CuAAC synthetic strategy. We selected glucose

(Glc) and glucosamine (GlcNAc), two rather ubiquitous

sugars, as the second recognition motif to confer hetero-

Scheme 1. Click synthesis of divalent heterogeneous neoglycoconju-

gates 12–15.

In a second strategy, we prepared divalent hetero neogly-

geneity. We prepared the alkynyl (2 and 3)^[32]and azido de- coconjugates with carbohydrate motifs connected by a

rivatives (4 and 5)^[33]of those sugars following reported pro- linker longer than that of the divalent systems 8 and 9 to

cedures. As denoted, we used all these clickable sugars as evaluate the influence of the distance between the sugar

peracetylated derivatives to facilitate their solubility in the moieties on the binding properties (Scheme 2). Thus, we

organic media required for the cycloaddition reactions (Fig- first reacted the alkynyl α-Man derivative 1 and the azide

ure 1). In general, we conducted the CuAAC reactions using Glc and GlcNAc derivatives 4 and 5 by CuAAC with 2-

the Cu catalyst (EtO)₃P·CuI (10 mol-%) in refluxing toluene azidoethanol and propargyl alcohol, respectively. In this

or toluene/THF (1:1) as the optimal conditions considering way, we obtained the corresponding glycosides containing

our previous successful contributions.^[26–28]

a 1,2,3-triazole-based aglycon bearing a terminal hydroxy

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Scheme 2. Modular click synthesis of divalent heterogeneous neoglycoconjugates.

group (17, 23, and 26) and transformed them into the azido an overall five-step sequence (orthogonal protection, func-

sugars 19, 25, and 28 by a two-step procedure: we obtained tionalization, click ligation, functionalization, click lig-

their chloride derivatives (18, 24, and 27) and subsequently ation) that allows the controllable sequential introduction

exposed them to nucleophilic substitution with sodium az- of the azide and/or the alkyne click functions onto those

ide. A second CuAAC reaction of those azido compounds scaffolds and their successive ligation with complementary

with the complementary functionalized alkynyl Glc deriva- functionalized carbohydrate derivatives. In this Figure, the

tive 3 (in the case of the α-Man-containing sugar 19) and two variants of this strategy used in the present study are

the alkynyl α-Man derivative 1 (in the case of the Glc- and indicated: sequential introduction of azide groups in the

GlcNAc-containing sugars 25 and 28, respectively) led to two functionalization steps (strategy A), and introduction,

the hetero divalent neoglycoconjugates 20, 32, and 34 pos- first, of alkyne groups followed by the subsequent introduc-

sessing a bis(triazolylmethyl) linker. The α-Man homogen- tion of azide groups after the first epitope click grafting

eous, divalent, neoglycoconjugate 34 was also synthesized (strategy B).

to be used as a customized reference in the binding essays

following an identical strategy starting from the azido α- galactopyranosides,^[35]and α,αЈ-trehalose polyhydroxylated

Man derivative 6. scaffolds. The advantages of carbohydrates over other syn-

We selected pentaerythritol,^[26,34]methyl α--gluco- and

We next undertook the synthesis of heterogeneous neo- thetic scaffolds include their availability in a variety of dia-

glycoconjugates with a valency higher that two. In order to stereomeric forms and their inherent polyfunctionality. In

develop a wide and flexible CuAAC-based strategy for the addition, some of them are among the cheapest enantiopure

synthesis of such compounds, we considered polyhydroxylic organic compounds available.^[10,36,37]On the other hand,

molecules as suitable scaffolds to be used in the modular the previous set of clickable alkynyl derivatives 1 and 3 and

assembly of sugar derivatives differing in their nature. These azides 4–7 was now expanded by including the 2-propynyl

scaffolds can be easily functionalized as azide and alkyne 2,3,4,6-tetra-O-acetyl-α--glucopyranoside (2), the α-ano-

derivatives. The strategy is depicted in Figure 2 and shows mer of 3 (Figure 1).

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Click Multivalent Heterogeneous Neoglycoconjugates

Figure 2. Modular assembly strategies for the synthesis of tetra- and octavalent heterogeneous neoglycoconjugates.

For the initial orthogonal protection of the scaffolds, we and 3, respectively, gave the hetero neoglycoconjugates 46

chose acetal derivatives in both of the strategies owing to and 47. In order to have model compounds as references

their easy preparation and the effectiveness of the required for the correct evaluation of the biological activity of such

ultimate hydrolytic deprotection. Thus, we prepared the compounds, we also reacted 44 and 45 with 2 and 1, respec-

pentaerythritol, Glc, and trehalose benzylidene derivatives tively, to give the Glc- and α-Man-containing homo-neogly-

38,^[37]59,^[38]and 67,^[38]respectively, and the isopropylidene coconjugates 48 and 49, respectively Similarly, we treated

Gal derivative 63^[39]following described methods in the lit- 43 with propargyl alcohol to give 50 (see Scheme 3).

erature. For the introduction of the azido function in those

We applied strategy B for the construction of pentaeryth-

partially protected scaffolds, alkylation with bis(2-chloro- ritol-centered hetero neoglycoconjugates as well as for the

ethyl) ether was followed by nucleophilic substitution with synthesis of their sugar-centered counterparts. In these

sodium azide (see Schemes 3 and 4). In this way, we effec- cases, the first functionalization step followed a similar

tively introduced clickable, medium-sized linkers into the pattern that in the strategy A with the only difference being

scaffolds, which are thereby prepared for the ultimate graft- that the first alkylation was performed with propargyl bro-

ing of complementary sugar appendages. We prepared the mide starting from the protected derivatives 38, 59, 63, and

alkynyl derivatives required in strategy B starting from the 67. We thus isolated the corresponding polyalkynylchloro

adequately protected hydroxyl scaffolds and by alkylation derivatives 58, 62, 66, and 70 as indicated in Scheme 4. We

with propargyl bromide (see Scheme 4).

then reacted these compounds with the azidoethyl α-Man

Using the above-indicated click reaction conditions, we derivative 7 giving the α-Man-containing chloro derivatives

prepared the pentaerythritol-centered hetero neoglycocon- 71, 77, 81, and 85. The exchange of the chlorine atom for

jugates 46 and 47 following strategy A. In this case, we per- the azido group and a further cycloaddition with the alk-

formed the first functionalization process starting from 38, ynyl derivatives 2 or 3 yielded the hetero neoglycoconju-

which comprised O-alkylation with bis(2-chloroethyl) ether, gates 73, 74, 79, 83, 87, and 88 (Scheme 5).

nucleophilic substitution of the chlorine atom by the azide

All the CuAAC reactions furnished the per-O-acetylated,

anion, hydrolysis of the acetal group, and, finally, a new multivalent, neoglycoconjugate derivatives in high yields.

alkylation reaction with bis(2-chloroethyl) ether giving ac- We performed de-O-protection of all these compounds

cess to the diazido-dichloro derivative 41. From this com- using NaOMe or Et₃N in MeOH in order to obtain the

pound the catalyzed cycloaddition reaction with the alkynyl corresponding hydroxylated compounds (12–15, 21, 35–37,

sugar 2 and 1 led to the Glc- and α-Man-containing chloro 51–55, 75–76, 60, 84, and 89–90) (see Schemes 1, 2, 3, 4,

derivatives 42 and 43, respectively. The reaction of these and 5) to be evaluated for their relative binding inhibitory

compounds with sodium azide allowed for the introduction properties against peroxidase-labeled Con A lectin. All the

of new azide groups and the isolation of 44 and 45, from new compounds were adequately characterized by spectro-

which the CuAAC with the complementary alkynyl sugar 1 scopic techniques.

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Scheme 3. Synthesis of pentaerythritol-centered hetero neoglycoconjugates. Strategy A.

Scheme 4. Preparation of functionalizated scaffolds for the synthesis of hetero neoglycoconjugates. Strategy B.

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Click Multivalent Heterogeneous Neoglycoconjugates

Scheme 5. Synthesis of pentaerythritol- and carbohydrate-centered hetero neoglycoconjugates. Strategy B.

Lectin Affinity Evaluation

to inhibit the lectin binding to a polymeric ligand coated

onto a microtiter well. In the present case, we carried out

We evaluated the affinities of the new hetero neoglycoconju- competitive experiments using horseradish peroxidase-lab-

gates toward Con A by the ELISA-type protocol ELLA. eled Con A (HRP-Con A) as the lectin and yeast mannan

This experiment measures the capacity of a soluble ligand as the microplate-fixed ligand. We also included methyl α-

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Man and the homo-neoglycoconjugates 14, 15, 37, 53, and evaluate the influence of such factors on carbohydrate-pro-

54 in the tests as reference compounds. We considered up tein recognition. The whole set of compounds present three

to six or seven different concentrations of each sample and valency values (divalent, tetravalent, and octavalent com-

determined the percentage of the inhibition of HRP-Con pounds), three different sugar substitution patterns for

A-yeast mannan association spectrophotometrically. We de- the hetero neoglycoconjugates (α-Man/β-Glc, α-Man/β-

termined the IC₅₀values, defined as the concentration of GlcNAc, and α-Man/α-Glc) and two for the homo-neogly-

synthetic compound required to achieve 50% inhibition of coconjugate (α-Man/α-Man and α-Glc/α-Glc), two different

this association, from the corresponding inhibition curves. 1,2,3-triazole-sugar grafting patterns (direct grafting to the

We assumed IC₅₀values to be inversely proportional to the anomeric position when 4, 5, and 6 are used and indirect

corresponding free energy of binding. The binding affinities grafting by means of an OCH₂or OCH₂CH₂tether when

of the references and the hetero neoglycoconjugates for Con 1–3 and 7 are clicked), and six different spacer arms [Tri-1,

A are summarized in Table 1 for the divalent compounds, Tri-2. Bis-Tri-1, and Bis-Tri-2 for the divalent compounds,

and in Table 2 for the multivalent ones. The valency-cor- and the OCH₂and (OCH₂CH₂)₂tether for the tetra- and

rected relative binding potencies are expressed per mol of octavalent derivatives] (see Table 2).

α-Man residue and per mol of sugar moiety relative to the

monovalent methyl α-Man.

In the case of the divalent neoglycoconjugates, several

conclusions can be extracted from the experimental data.

As the new homo and hetero neoglycoconjugates display Firstly, the binding affinities are a function of the distance

structural differences that are governed by the valency, scaf- between the sugar residues. This we observed in the hetero

fold architecture, sugar substitution, configurational (α or neoglycoconjugates 12–13 and 35–36 having the same sub-

β) pattern, grafting pattern, and spacer length, we could stitution (α-Man/β-GlcNAc) and grafting pattern. In these

Table 1. Binding affinities toward Con A for the divalent hetero neoglycoconjugates.

Relative

affinity

Relative potency

per α-Man moiety

Relative potency per

sugar moiety

Entry

Compound

Core

Sugar A

Sugar B

IC₅₀[m]

1

2

3

4

5

6

7

8

9

Methyl α--Man

0.9

1

12

13

14

15

21

35

36

37

Tri-1

α-Man

β-Glc

β-GlcNAc

α-Man

β-Glc

β-GlcNAc

α-Man

0.80

0.66

0.55

0.41

0.52

0.94

1.02

0.64

1.13

1.36

1.64

2.19

1.73

0.96

0.88

1.41

1.13

1.36

0.82

1.10

1.73

0.96

0.88

0.71

0.56

0.68

0.82

1.10

0.86

0.48

0.44

0.71

Tri-2

Bis-Tri-1

Bis-Tri-2

Table 2. Binding affinities toward Con A for the tetravalent and octavalent hetero neoglycoconjugates .

Relative

affinity

Relative potency per Relative potency per

Entry

Compound

Core

Sugar A

Sugar B

IC₅₀[m]

α-Man moiety

sugar moiety

1

2

3

4

5

6

7

8

Methyl α--Man

0.90

0.21

0.48

0.16

0.20

0.23

0.39

0.40

0.37

0.087

0.30

1

51

52

53

54

55

75

76

80

84

89

90

Ery-l

Ery-s

Glc

α-Man

α-Glc

β-Glc

α-Glc

α-Man

OH

α-Glc

β-Glc

α-Glc

β-Glc

4.28

1.87

5.62

4.50

3.91

2.30

2.25

2.43

10.34

3

2.14

–

1.07

0.47

1.40

2.25

0.98

0.57

0.56

0.60

1.29

0.37

α-Man

1.40

2.25

1.95

1.15

1.12

1.21

2.58

0.75

9

10

11

12

Gal

Thr

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Click Multivalent Heterogeneous Neoglycoconjugates

compounds, the relative binding affinities per mol of α-Man fering in structural factors such as the valency, scaffold

residue are higher for 12 and 13 (1.13 and 1.36), where the architecture, anomeric configuration (α or β) of these

sugar residues are connected through the Tri-1 spacer arm, sugars, grafting pattern, and the length of the spacer that

than they are for their counterparts 35 and 36 (0.96 and links the sugars to the core has been performed by means

0.88, valency-corrected affinity values, respectively) where of a series of different modular click-based strategies. The

the sugar residues are connected through the longer Bis-Tri- flexibility and efficiency of click chemistry made the re-

2 arm. We also observed this favorable effect for the homo ported methodology a highly versatile and simple tool for

divalent 14 and 37 (0.82 vs. 0.71). Secondly, the affinity val- accessing well-defined and custom-made, multivalent,

ues are also a function of the grafting pattern. Thus, we heterogeneous neoglycoconjugates. All the heterogeneous

observed a higher relative potency per sugar moiety in both neoglycoconjugates synthesized were α-Man-containing

the homo and hetero neoglycoconjugates when the sugar glycomimics, and assays to evaluate their binding properties

was not directly connected through its anomeric position to toward the natural carbohydrate binding lectin Con A have

the 1,2,3-triazole ring; compare 15 (Tri-2 spacer) and 21 allowed us to extract some conclusions about the influence

(Bis-Tri-1 spacer) with their counterparts 12–14 (Tri-1 of the above-mentioned structural factors on the inhibitory

spacer) and 35–37 (Bis-Tri-2 spacer). We obtained these properties of these novel compounds. Remarkably, the sub-

compounds when we used the azide glycosides 4–6 in click stitution pattern and the distance between the sugar are the

ligations, and these results point to a disfavored accessibility more important parameters influencing the binding capa-

by the lectin receptors for the sugar residues owing to steric bilities of these compounds.

hindrance.

Concerning the tetravalent and octavalent neoglycocon-

Experimental Section

jugates, we first observed that in compounds having the α-

Man/β-Glc substitution pattern connected to the core by

identical spacer arms (76, 80, 84, and 90) there is neither

an influence of the core, as 76, 80, and 84 have similar rela-

tive affinity values, nor an influence of the valency, as the

General: TLC was performed on Merck silica gel 60 F₂₅₄aluminum

sheets. Reagents used for developing plates include ceric sulfate

(1% w/v) and ammonium sulfate (2.5% w/v) in 10% (v/v) aqueous

sulfuric acid, iodine, ethanolic sulfuric acid (10% v/v), and UV

observed increase in the relative affinity value for 90 is not light when applicable. Flash column chromatography was per-

formed on Silica Gel Merck (230–400 mesh, ASTM). Melting

points were measured with a Gallenkamp melting point apparatus

and are uncorrected. Optical rotations were recorded with a Per-

kin–Elmer 141 polarimeter at room temp. IR spectra were recorded

with a Satellite Mattson FTIR spectrometer. ¹H and ¹³C NMR

spectra were recorded at room temp. with a Bruker (300 or

400 MHz) spectrometer. Chemical shifts are given in ppm and ref-

erenced to internal CDCl₃. J values are given in Hz. Assignments

were based on COSY, HMQC, NOESY, and DEPT experiments.

FAB mass spectra were recorded with a Fissons VG Autospec-Q

indicative of a cluster effect. However, we did observe a

cooperative effect, as expected, for those compounds

wherein the anomeric configuration of the Glc moiety was

inverted (α-Man/α-Glc substitution pattern); compare 75

and 89 (3.91 and 10.34 relative affinity values) with their

counterparts 76 and 90 (2.30 and 3.0 relative affinity val-

ues). On the other hand, when we compared the ensemble

of the pentaerythritol hetero derivatives (51, 52, 55, 75, and

76) in order to preclude the influence of the core, we ob-

served that the main factor that contributes to the inhibi- spectrometer, using m-nitrobenzyl alcohol or thioglycerol as the

matrix. Matrix-assisted laser desorption/ionization time-of-flight

mass spectrometry (MALDI-TOF MS) was performed with a

Bruker Daltonics (AUTOFLEX) spectrometer using DGB as the

matrix. Compounds 1–7, 38, 59, 63, and 67 were obtained follow-

ing the described methods in the literature.^{[30–33,37–39]}

tory properties of these compounds is the length of the

spacer. Thus, 51, 52, and 55 with the (OCH₂CH₂)₂spacer

have higher affinity values than do their homologous 75

and 76 with the OCH₂spacer irrespective of the nature of

the second ligand. This fact points to the idea that only the

α-Man residues are determinant in the carbohydrate-pro-

tein recognition because they have the adequate spatial ar-

rangement to interact with the binding site of Con A. This

is supported by the slight increase in the affinity value that

we observed for the α-Man homo neoglycoconjugate 54

when compared with the rest of the pentaerythritol hetero

derivatives bearing the same (OCH₂CH₂)₂spacer (51, 52,

and 55). Finally, we note that the homo neoglycoconjugates

General Procedure for the Propargylation of Hydroxylated Deriva-

tives: To a solution of the corresponding hydroxylated derivative

(1 mmol) in dry DMF (5 mL) was added NaH (4 mmol). After the

reaction mixture was stirred for 0.5 h under an argon atmosphere,

propargyl bromide (6 mmol) was added, and the reaction was

maintained at room temp. for 16 h. MeOH (5 mL) was then added,

and the solvents were evaporated. The crude material was dissolved

in toluene/diethyl ether (100 mL, 1:1) and washed with H₂O. The

organic phase was dried, concentrated, and purified by column

53 and 54 showed the expected behavior, with a three-fold chromatography (diethyl ether/hexane, 1:1).

increase in the relative affinity of the α-Man derivative rela-

General Procedure for Acetal Hydrolysis: A solution of the corre-

tive to that of its α-Glc counterpart.

sponding acetal derivative (1 mmol) in AcOH/H₂O (7:3, 10 mL)

was heated at 50 °C for 2 h. Evaporation of the solvent gave a crude

material that was purified by column chromatography.

General Procedure for the Synthesis of Chloro Derivatives: A solu-

tion of the corresponding hydroxylated sugar derivative (1 mmol),

SOCl₂(0.12 mL, 1.7 mmol) and pyridine (0.15 mL, 1.8 mmol) in

Conclusions

In summary, the synthesis of a variety of multivalent

neoglycoconjugates possessing two different sugars and dif- THF (15 mL) was keep at room temp. for 30 min. The reaction

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mixture was evaporated and the resultant crude product was puri-

fied by column chromatography.

azole (9): Obtained from 1 (386 mg) and 5 (446 mg) following the

general procedure for the CuAAC reactions. Column chromatog-

20

raphy (EtOAc) gave 9 as a solid (690 mg, 91%); m.p. 80 °C. [α]

436

General Procedure for the Synthesis of (2-Chloroethoxy)ethyl Deriv-

atives: A solution of the corresponding hydroxylated derivatives

(1 mmol) and tetrabutylammonium hydrogensulfate (2 mmol) in

bis(2-chloroethyl) ether (5 mL) was vigorously stirred at room

temp. with 50% aq NaOH (5 mL) for 24 h. The reaction mixture

was diluted in H₂O and CH₂Cl₂, the phases were separated, and

the aqueous phase was washed with CH₂Cl₂. The organic extracts

were combined, washed with H₂O, dried, and concentrated under

vacuum. The resultant crude product was purified by column

chromatography.

= +14 (c = 1 in chloroform). IR (KBr): ν = 2955, 1748, 1371, 1230,

˜

1042 cm^–1. ¹H NMR (CDCl₃, 300 MHz): δ = 7.97 (s, 1 H, H-5

triazole), 6.82 (d, J = 9.1 Hz, 1 H, NH), 6.19 (d, J = 10 Hz, 1 H,

H-1 GlcNAc), 5.59 (t, J = 9.9 Hz, 1 H, H-3 GlcNAc), 5.32–5.22

(m, 4 H, H-4 GlcNAc, H-2,3,4 Man), 4.95 (s, 1 H, H-1 Man), 4.85

(d, J = 12.3 Hz, 1 H, CH₂O), 4.73 (d, J = 12.3 Hz, 1 H, CH₂O),

4.54 (q, J = 9.9 Hz, 1 H, H-2 GlcNAc), 4.36–4.28 (m, 2 H, H-6

GlcNAc, H-6 Man), 4.19–4.09 (m, 4 H, H-5,6’ GlcNAc, H-5,6’

Man), 2.15, 2.13, 2.09, 2.07, 2.04, 1.98, 1.78 (7 s, 24 H, 8 Ac) ppm.

¹³C NMR (CDCl₃, 75 MHz): δ = 170.7, 170.6, 170.0, 169.9, 169.7,

169.3 (CO), 143.7 (C-4 triazole), 122.4 (C-5 triazole), 96.8 (C-1

Man), 85.7 (C-1 GlcNAc), 74.8, 72.1, 69.5, 69.0, 68.7, 68.2, (C-

2,3,5 Man, C-3,4,5 GlcNAc), 65.9 (C-4 Man), 62.3, 61.7 (C-6 Man,

C-6 GlcNAc), 60.7 (CH₂O), 53.6 (C-2 GlcNAc), 22.7 (MeCON),

20.8, 20.7, 20.6 (MeCO) ppm. HRMS (FAB+): calcd. for

C₃₁H₄₂N₄O₁₈[M + Na]⁺781.2392; found 781.2398.

General Procedure for the Synthesis of the Azides: A solution of the

corresponding (2-chloroethoxy)ethyl derivative (1 mmol), tetrabu-

tylammonium iodide (0.01 mmol), and sodium azide (5 mmol) in

dry DMF (7 mL) was heated at 75–80 °C for 20 h. After the mix-

ture was cooled, the crude material was dissolved in toluene/diethyl

ether (100 mL, 1:1) and washed with H₂O. The organic phase was

dried, concentrated, and purified by column chromatography.

1-(2,3,4,6-Tetra-O-acetyl-α-

D-mannopyranosyl)-4-(2,3,4,6-tetra-O-

General Procedure for the CuAAC Reactions: The corresponding

alkyne or azide (1 equiv.), was treated with the azido α-Man deriva-

tive 4, 5, 6, or 7 or the propargyl glycoside 1–3 or propargyl alcohol

(1.2 equiv.), respectively, using the copper catalyst (EtO)₃P·CuI

(10 mol-%) in refluxing toluene or toluene/THF (1:1) for 20–30 min

until TLC showed complete disappearance of the limiting reagent.

The reaction mixture was concentrated, and the crude material was

purified by short-column flash chromatography.

acetyl-α- -mannopyranosyloxy)-1H-1,2,3-triazole (10): Obtained

D

from 1 (386 mg) and 6 (448 mg) following the general procedure

for the CuAAC reactions. Column chromatography (EtOAc/hex-

ane, 3:1) gave 10 as a solid (698 mg, 92%); m.p. 60 °C. [α]²_D⁰= +54

(c = 1 in chloroform). IR (KBr): ν = 2937, 1746, 1366, 1221, 1039

˜

cm^–1. ¹H NMR (CDCl₃, 300 MHz): δ = 7.83 (s, 1 H, H-5 triazole),

6.06 [d, J = 2.8 Hz, 1 H, H-1 Man(N)], 5.97 [t, J = 3.3 Hz, 1 H,

H-2 Man(N)], 5.92 [dd, J = 8.7, 3.7 Hz, 1 H, H-3 Man(N)], 5.39

[t, J = 8.7 Hz, 1 H, H-4 Man(N)], 5.33–5.30 [m, 2 H, H-3,4

Man(O)], 5.25 [br. s, 1 H, H-2 Man(O)], 4.97 [d, J = 1.2 Hz, 1 H,

H-1 Man(O)], 4.90 (d, J = 12.4 Hz, 1 H, CH₂O), 4.73 (d, J =

12.3 Hz, 1 H, CH₂O), 4.42 [dd, J = 12.5, 5.3 Hz, 1 H, H-6 Man

(N)], 4.31 [dd, J = 12.5, 5.3 Hz, 1 H, H-6’ Man(N)], 4.16–4.07 [m,

3 H, H-5,6,6’ Man(O)], 3.96 [ddd, J = 8.9, 5.3, 2.6 Hz, 1 H, H-5

Man(N)], 2.18, 2.16, 2.13, 2.10, 2.08, 2.06, 2.04, 1.99 (8 s, 24 H, 8

Ac) ppm. ¹³C NMR (CDCl₃, 75 MHz): δ = 170.6, 170.5, 169.9,

169.8, 169.7, 169.6, 169.2 (CO), 144.2 (C-4 triazole), 123.4 (C-5

triazole), 97.0 [C-1 Man(O)], 83.6 [C-1 Man(N)], 72.3 [C-5

Man(N)], 69.4, 69.0, 68.8, 68.7, [C-2,3,5 Man(O), C-3 Man(N)],

68.2 [C-2 Man(N)], 66.1 [C-4 Man(N), C-4 Man(O)], 62.3 [C-6

Man(O)], 61.5 [C-6 Man(N)], 60.7 (CH₂O), 20.8, 20.7, 20.6, 20.5

(MeCO) ppm. HRMS (FAB+): calcd. for C₃₁H₄₁N₃O₁₉[M +

Na]⁺782.2232; found 782.2238.

General Procedure for the De-O-Acetylation of Per-O-acetylated

Multivalent Neoglycoconjugates. Method A: A solution of the corre-

sponding per-O-acetylated neoglycoconjugate (0.1 mmol) in

MeOH (8 mL) containing Et₃N (0.8 mL) was maintained at 50 °C

for 2–3 h. The evaporation of the solvent was followed by purifica-

tion of the crude material by short-column flash chromatography

(MeOH) column.

Method B: To a solution of the corresponding per-O-acetylated

neoglycoconjugate (0.1 mmol) in MeOH (15 mL) was added Na-

OMe (1 , 0.1 mL). After 2 h, the solution was treated with Am-

berlite IR-120(H⁺), filtered, and the solvent was evaporated. The

resulting crude material was purified by short-column flash

chromatography.

1-(2,3,4,6-Tetra-O-acetyl-β-D-glucopyranosyl)-4-(2,3,4,6-tetra-O-ace-

tyl-α- -mannopyranosyloxymethyl)-1H-1,2,3-triazole (8): Obtained

D

from 1 (386 mg) and 4 (448 mg) following the general procedure

1-(2,3,4,6-Tetra-O-acetyl-α-

D-mannopyranosyloxyethyl)-4-(2,3,4,6-

for the cycloaddition reactions. Column chromatography (EtOAc/

tetra-O-acetyl-α- -mannopyranosyloxymethyl)-1H-1,2,3-triazole

D

20

hexane, 5:2) gave 8 as a solid (706 mg, 93%); m.p. 93 °C. [α]

=

436

(11): Obtained from 1 (386 mg) and 7 (500 mg) following the gene-

+23 (c = 1 in chloroform). IR (KBr) ν = 2952, 1747, 1368, 1324,

˜

ral procedure for the CuAAC reactions. Column chromatography

1037 cm^–1. ¹H NMR (CDCl₃, 300 MHz): δ = 7.86 (s, 1 H, H-5

triazole), 5.86 (m, 1 H, H-1 Glc), 5.46–5.38 (m, 2 H, H-2,3 Glc),

5.34–5.22 (m, 4 H, H-4 Glc, H-2,3,4 Man), 4.22 (br. s, 1 H, H-1

Man), 4.85 (d, J = 12.1 Hz, 1 H, CH₂O), 4.68 (d, J = 12.1 Hz, 1

H, CH₂O), 4.31 (dd, J = 12.6, 5.0 Hz, 1 H, H-6 Glc), 4.18–4.08

(m, 4 H, H-6Ј Glc, H-5,6,6Ј Man), 4.00 (ddd, J = 10.1, 4.9, 2.1 Hz,

1 H, H-5 Glc), 2.13, 2.11, 2.09, 2.06, 2.02, 1.97, 1.86 (7 s, 24 H, 8

Ac) ppm. ¹³C NMR (CDCl₃, 75 MHz): δ = 170.4, 169.7, 169.6,

169.5, 169.1, 168.7 (CO), 144.1 (C-4 triazole), 121.5 (C-5 triazole),

96.7 (C-1 Man), 85.5 (C-1 Glc), 74.9 (C-5 Glc), 72.4, 70.2 (C-2,3

Glc), 69.2, 68.9, 68.6, 67.6 (C-2,3,5 Man, C-4 Glc), 65.9 (C-4 Man),

62.2, 61.4 (C-6 Man, C-6 Glc), 60.6 (CH₂O), 20.6, 20.6, 20.5, 20.3,

19.9 (MeCO) ppm. HRMS (FAB+): calcd. for C₃₁H₄₁N₃O₁₉[M +

Na]⁺782.2232; found 782.2230.

(EtOAc/hexane, 3:1) gave 11 as a syrup (802 mg, 92%): [α]²_D⁰

=

+37.5 (c = 1 in chloroform) IR (film): ν = 2750, 1436, 1369, 1234

˜

cm^–1. ¹H NMR (CDCl₃, 300 MHz): δ = 7.79 (s, 1 H, H-5 triazole),

65.35–5.18 (m, 6 H), 5.00 (s, 1 H), 4.88 (dd, J = 12.2 Hz, 1 H), 4.61

(d, J = 12.3 Hz, 1 H), 4.82 (s, 1 H), 4.64 (t, J = 5.0 Hz, 2 H), 4.32

(dd, J = 12.5, 5.1 Hz, 1 H), 4.21 (dd, J = 12.3, 5.1 Hz,1 H), 4.16–

4.10 (m, 3 H), 4.06 (dd, J = 12.4, 3.1 Hz, 1 H), 3.91 (dt, J = 10.7,

5.0 Hz, 1 H), 3.59 (m, 1 H), 2.15, 2.14, 2.21, 2.10, 2.05, 2.04, 2.00,

1.97 (8 s, 24 H) ppm. ¹³C NMR (CDCl₃, 75 MHz): δ = 170.5,

170.3, 169.8, 169.6, 169.5, 169.4, 143.5, 124.3, 97.2, 96.9, 69.3, 69.0,

68.9, 68.6, 68.5, 66.0, 65.6, 62.2, 62.1, 60.7, 49.6, 20.7, 20.6, 20.5

ppm. HRMS (FAB+): calcd. for C₃₃H₄₅N₃O₂₀[M + Na]⁺

826.2494; found 826.2491.

1-(2-Acetamido-2-deoxy-3,4,6-tri-O-acetyl-β-

D

-glucopyranosyl)-4-

1-(β-D-Glucopyranosyl)-4-(α-D-mannopyranosyloxymethyl)-1H-

(2,3,4,6-tetra-O-acetyl-α- -mannopyranosyloxymethyl)-1H-1,2,3-tri- 1,2,3-triazole (12): Obtained from 8 (759 mg) following the general

D

2462

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Eur. J. Org. Chem. 2009, 2454–2473

Click Multivalent Heterogeneous Neoglycoconjugates

procedure for the de-O-acetylation (method A). Column

chromatography (EtOAc/MeOH, 1:1) gave 12 as a solid (402 mg,

Column chromatography (EtOAc) gave 18 as a syrup (447 mg,

91%): [α]²_D⁰= +43 (c = 1 in chloroform). IR (film): ν = 2927, 1746,

˜

95%); m.p. 156–158 °C. [α]²_D⁰= +39 (c = 1 in H O). IR (KBr): ν =

1226, 1047 cm^–1. ¹H NMR (CDCl₃, 300 MHz): δ = 7.75 (s, 1 H,

˜

2

3386, 1641, 1560, 1455, 1408, 1235, 1126, 1047 cm^–1. ¹H NMR H-5 triazole), 5.32–5.30 (m, 2 H, H-3,4), 5.25 (s, 1 H, H-2), 4.97

(CD₃OD, 300 MHz, selected signals): δ = 8.25, (s, 1 H), 5.63 (d, J =

(s, 1 H, H-1), 4.87 (d, J = 12.4 Hz, 1 H, OCH₂-triazole), 4.74–4.70

9.1 Hz, 1 H) ppm. ¹³C NMR (CD₃OD, 75 MHz, selected signals): δ (m, 3 H, CH₂Cl, OCH₂-triazole), 4.30 (dd, J = 12.2, 5.0 Hz, 1 H,

= 145.3, 124.7, 100.8, 89.5, 81.0, 78.4, 74.8, 73.9, 72.4, 71.9, 70.8, H-6), 4.14–4.05 (m, 2 H, H-5,6’), 3.96 (t, J = 5.8 Hz, 2 H, CH₂N),

68.6, 62.9, 62.3, 60.7 ppm. HRMS (FAB+): calcd. for C₁₅H₂₅N₃O₁₉

2.15, 2.12, 2.04, 1.99 (4 s, 12 H, 4 Ac) ppm. ¹³C NMR (CDCl₃,

75 MHz): δ = 170.7, 170.1, 169.9 (CO), 143.5 (C-4 triazole), 124.1

(C-5 triazole), 96.9 (C-1), 69.5, 69.0, 68.8 (C-2,3,5), 66.1 (C-4), 62.4

(C-6), 61.0 (CH₂O), 51.8 (CH₂N), 42.4 (CH₂Cl), 20.9, 20.8, 20.7

(MeCO) ppm. HRMS (FAB+): calcd. for C₁₉H₂₆ClN₃O₁₀[M +

Na]⁺514.1204; found 514.1207.

[M + Na]⁺446.1386; found 446.1386.

1-(2-Acetamido-2-deoxy-β-D-glucopyranosyl)-4-(α-D-mannopyranos-

yloxymethyl)-1H-1,2,3-triazole (13): Obtained from 9 (758 mg) fol-

lowing the general procedure for the de-O-acetylation (method A).

Column chromatography (EtOAc/MeOH, 1:1) gave 13 as a foamy

solid (431 mg, 93%): [α]²_D⁰= +31 (c = 1 in MeOH). IR (KBr): ν =

˜

1-(2-Azidoethyl)-4-(2,3,4,6-tetra-O-acetyl-a-D-mannopyranosyloxy-

3365, 1657, 1550, 1376, 1316, 1238, 1098 cm^–1. ¹H NMR (CD₃OD,

300 MHz, selected signals): δ = 8.24 (s, 1 H), 5.82 (d, J = 9.7 Hz,

1 H), 1.79 (s, 3 H) ppm. ¹³C NMR (CD₃OD, 75 MHz, selected

signals): δ = 173.6, 145.3, 124.2, 100.3, 88.0, 81.0, 75.7, 74.8, 72.4,

71.9, 71.2, 68.6, 62.9, 62.2, 60.4, 56.7, 25.3 ppm. HRMS (FAB+):

calcd. for C₁₇H₂₈N₄O₁₁[M + Na]⁺487.16.52; found 487.1654.

methyl)-1H-1,2,3-triazole (19): Obtained from 18 (491 mg) follow-

ing the general procedure for the synthesis of the azido derivatives.

Column chromatography (EtOAc) gave 19 as a syrup (504 mg,

74%): [α]²_D⁰= +44 (c = 1 in chloroform). IR (film): ν = 2103, 1747,

˜

1

1361, 1226, 1045 cm^–1. H NMR (CDCl₃, 300 MHz): δ = 7.71 (s,

1 H, H-5 triazole), 5.36–5.30 (m, 2 H, H-3,4), 5.25 (br. s, 1 H, H-

2), 4.96 (br. s, 1 H, H-1), 4.87 (d, J = 12.4 Hz, 1 H, OCH₂-triazole),

4.72 (d, J = 12.4 Hz, 1 H, OCH₂-triazole), 4.53 (t, J = 5.6 Hz, 2

H, CH₂N), 4.30 (dd, J = 12.2, 5.1 Hz, 1 H, H-6), 4.15–4.07 (m, 2

H, H-5,6’), 3.85 (t, J = 5.6 Hz, 2 H, CH₂-triazole), 2.15, 2.12, 2.04,

1.99 (4 s, 12 H, 4 Ac) ppm. ¹³C NMR (CDCl₃, 75 MHz): δ = 170.7,

170.1, 170.0, 169.7 (CO), 143.9 (C-4 triazole), 123.9 (C-5 triazole),

96.9 (C-1), 69.5, 69.1, 68.8 (C-2,3,5), 67.2 (C-4), 62.4 (C-6), 61.0

(CH₂O), 50.7 (CH₂N), 49.5 (CH₂N₃), 20.9, 20.8, 20.7 (MeCO)

ppm. HRMS (FAB+): calcd. for C₁₉H₂₆N₆O₁₀[M + Na]⁺

521.1608; found 521.1607.

1-(α-D-Mannopyranosyl)-4-(α-D-mannopyranosyloxymethyl)-1H-

1,2,3-triazole (14): Obtained from 10 (759 mg) following the general

procedure for the de-O-acetylation (method A). Column

chromatography (EtOAc/MeOH, 1:1) gave 14 as a syrup (368 mg,

87%): [α]²_D⁰= +62 (c = 1 in H O). IR (film): ν = 3393, 1643, 1413,

˜

2

1346, 1235, 1124, 1122 cm^–1. ¹H NMR (CD₃OD, 300 MHz, se-

lected signals): δ = 8.21 (s, 1 H), 6.04 (d, J = 2.7 Hz) ppm. ¹³C

NMR (CD₃OD, 75 MHz, selected signals): δ = 145.6, 125.3, 100.9,

88.3, 78.6, 74.9, 72.5, 72.4, 71.9, 70.0, 68.6, 68.6, 62.9, 62.5, 60.7

ppm. HRMS (FAB+): calcd. for C₁₅H₂₅N₃O₁₁[M + Na]⁺

446.1386; found 446.1376.

2-[4-(2,3,4,6-Tetra-O-acetyl-β-

D-glucopyranosyloxymethyl)-1-[4-

1-(α-D-Mannopyranosyloxyethyl)-4-(α-D-mannopyranosyloxymeth-

(2,3,4,6-tetra-O-acetyl-α- -mannopyranosyloxymethyl)-1H-1,2,3-tri-

D

yl)-1H-1,2,3-triazole (15): Obtained from 11 (803 mg) following the

general procedure for the de-O-acetylation (method A). Column

chromatography (EtOAc/MeOH, 1:2) gave 15 as a foamy solid

azol-1-yl]-1H-1,2,3-triazol-1-yl]ethane (20): Obtained from 3

(463 mg) and 19 (498 mg) following the general procedure for the

CuAAC reactions. Column chromatography (EtOAc) gave 20 as a

(453 mg, 97%): [α]²_D⁰= +63 (c = 1 in H O). IR (KBr): ν = 3400,

20

syrup (822 mg, 93%): [α]²_D⁰= +5, [α] = +23 (c = 1 in chloroform).

˜

2

436

1641, 1133, 1056 cm^–1. H NMR (CD₃OD, 300 MHz, selected sig-

1

IR (film): ν = 2103, 1750, 1370, 1226, 1043 cm^–1. ¹H NMR (CDCl ,

˜

3

nals): δ = 8.06 (s, 1 H), 4.77 (s, 1 H), 4.72 (s, 1 H) ppm. ¹³C NMR

(CD₃OD, 75 MHz, selected signals): δ = 143.9, 124.9, 100.4, 99.3,

73.7, 73.6, 71.2, 71.2, 70.7, 70.5, 67.3, 67.0, 65.5, 61.6, 61.5, 59.2,

49.9 ppm. HRMS (FAB+): calcd. for C₁₇H₂₉N₃O₁₂[M + Na]⁺

490.1649; found490.1649.

300 MHz): δ = 7.45, 7.43 (2 s, 2 H, H-5 triazole), 5.31–5.28 (m, 2

H, H-3,4 Man), 5.21 (t, J = 9.4 Hz, 1 H, H-3 Glc), 5.20 (br. s, 1

H, H-2 Man), 5.09 (t, J = 9.6 Hz, 1 H, H-4 Glc), 4.98 (dd, J = 9.6,

8.0 Hz, 1 H, H-2 Glc), 4.95 (br. s, 4 H, 2 CH₂N), 4.94 (d, J =

1.7 Hz, 1 H, H-1 Man), 4.89 (d, J = 12.6 Hz, 1 H, CH₂O), 4.77 (d,

J = 12.6 Hz, 1 H, OCH₂), 4.80 (d, J = 12.4 Hz, 1 H, CH₂O), 4.64

(d, J = 12.4 Hz, 1 H, CH₂O), 4.65 (d, J = 7.9 Hz, 1 H, H-1 Glc),

4.29 (dd, J = 12.3, 5.0 Hz, 1 H, H-6 Man), 4.26 (dd, J = 12.3,

4.4 Hz, 1 H, H-6 Glc), 4.19–4.09 (m, 2 H, H-6’ Glc, H-6’ Man),

4.06 (ddd, J = 9.7, 5.0, 2.4 Hz, 1 H, H-5 Man), 3.74 (ddd, J = 9.9,

4.5, 2.4 Hz, 1 H, H-5 Glc), 2.15, 2.12, 2.09, 2.05, 2.04, 2.03, 2.00,

1.98 (8 s, 24 H, 4 Ac) ppm. ¹³C NMR (CDCl₃, 75 MHz): δ = 170.6,

169.9, 169.7, 169.2 (CO), 144.4, 143.8 (C-4 triazole), 124.1 (C-5

triazole), 100.0 (C-1 Glc), 96.8 (C-1 Man), 72.7, 71.9, 71.1, 69.4,

69.0, 68.7, 68.3 (C-2,3,5 Glc, C-2,3,5 Man), 66.0 (C-4 Man), 62.8,

62.3 (C-6 Glc, C-6 Man), 61.7, 60.7 (2 CH₂O), 49.5, 49.4 (2 CH₂N),

20.8, 20.7, 20.6, 20.5 (MeCO) ppm. HRMS (FAB+): calcd. for

C₃₆H₄₈N₆O₂₀[M + Na]⁺907.2821; found 907.2833.

1-(2-Hydroxyethyl)-4-(2,3,4,6-tetra-O-acetyl-a-D-mannopyranosyl-

oxymethyl)-1H-1,2,3-triazole (17): Obtained from 1 (386 mg) and

16 (104 mg) following the general procedure for the CuAAC reac-

tions. Column chromatography (EtOAcǞ EtOAc/MeOH, 10:1)

gave 17 as a syrup (435 mg, 92%): [α]²_D⁰= +43 (c = 1 in chloroform).

IR (film): ν = 2940, 1746, 1369, 1226, 1047 cm^–1. ¹H NMR (CDCl ,

˜

3

300 MHz): δ = 7.79 (s, 1 H, H-5 triazole), 5.32, 5.29 (m, 2 H, H-

3,4), 5.20 (s, 1 H, H-2), 5.96 (s, 1 H, H-1), 4.84 (d, J = 12.4 Hz, 1

H, OCH₂-triazole), 4.7 (d, J = 12.4 Hz, 1 H, OCH₂triazole), 4.51

(d, J = 4.9 Hz, 2 H, CH₂OH), 4.27 (dd, J = 12.3, 5.0 Hz, 1 H, H-

6), 4.14–4.04 (m, 4 H, H-5,6’, CH₂N), 3.66 (br. s, 1 H, OH), 2.15,

2.11, 2.04, 1.98 (4 s, 12 H, 4 Ac) ppm. ¹³C NMR (CDCl₃, 75 MHz):

δ = 170.2, 170.0, 169.7 (CO), 143.2 (C-4 triazole), 124.3 (C-5 tri-

azole), 96.9 (C-1), 69.5, 69.1, 68.7 (C-2,3,5), 66.1 (C-4), 62.4 (C-6),

61.1, 61.0 (OCH₂-triazole, CH₂OH), 52.8 (CH₂N), 20.9, 20.8, 20.7

(MeCO) ppm. HRMS (FAB+): calcd. for C₁₉H₂₇N₃O₁₁[M +

Na]⁺496.1543; found 496.1539.

2-[4-(β-

D-Glucopyranosyloxymethyl)-1H-1,2,3-triazol-1-yl]-1-[4-(α-

D-mannopyranosyloxymethyl)-1H-1,2,3-triazol-1-yl]ethane (21): Ob-

tained from 20 (884 mg) following the general procedure for the

de-O-acetylation (method A). Column chromatography (EtOAc/

1-(2-Chloroethyl)-4-(2,3,4,6-tetra-O-acetyl-α-D

-mannopyranosyloxy- MeOH, 1:1) gave 21 as a syrup (537 mg, 98%): [α]²_D⁰= +21 (c = 1

methyl)-1H-1,2,3-triazole (18): Obtained from 17 (473 mg) follow-

ing the general procedure for the synthesis of chloro derivatives.

in H O). IR (KBr): ν = 3394, 1644, 1367, 1230, 1125, 1054 cm^–1.

˜

2

¹H NMR (CD₃OD, 300 MHz): δ = 7.87 (s, 1 H, H-5 triazole), 7.80

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F. Santoyo-Gonzalez et al.

FULL PAPER

(s, 1 H, H-5 triazole), 4.92 (d, J = 12.7 Hz, 1 H), 4.78 (s, 1 H), 170.9, 170.6, 169.4, 148.1, 121.3, 85.9, 74.8, 72.5, 68.2, 61.8, 56.1,

4.766 (d, J = 12.9 Hz, 1 H), 4.75 (d, J = 12.5 Hz, 1 H), 4.61 (d, J = 53.5, 22.7, 20.7, 20.7, 20.6 ppm. HRMS (FAB+): calcd. for

12.5 Hz, 1 H), 4.34 (d, J = 7.8 Hz, 1 H) ppm. ¹³C NMR (CD₃OD, C₁₇H₂₄N₄O₉[M + Na]⁺451.1441; found 451.1437.

75 MHz): δ = 144.7, 144.2, 124.6, 124.5, 102.2, 99.3, 76.7, 73.7,

1-(2-Acetamido-2-deoxy-3,4,6-tri-O-acetyl-β-D-glucopyranosyl)-4-

71.1, 70.6, 70.3, 67.3, 61.6, 61.5, 61.4, 59.2, 49.6 ppm. HRMS

(FAB+): calcd. for C₂₀H₃₂N₆O₁₂[M + Na]⁺571.1976; found

571.1970.

(chloromethyl)-1H-1,2,3-triazole (27): Obtained from 26 (428 mg)

following the general procedure for the synthesis of chloro deriva-

tives. Column chromatography of the crude material (EtOAc/

MeOH, 20:1) gave 27 as a solid (448 mg, 90%); m.p. 210 °C (dec).

4-(Hydroxymethyl)-1-(2,3,4,6-tetra-O-acetyl-β-D-glucopyranosyl)-

1H-1,2,3-triazole (23): Obtained from 4 (373 mg) and 22 (67 mg)

following the general procedure for the CuAAC reactions.

Crystallization of the crude material (diethyl ether/hexane) gave 23

as a solid (420 mg, 98%); m.p. 156–158 °C. [α]²_D⁰= –18 (c = 1 in

[α]²_D⁰= –42 (c = 1 in chloroform). IR (KBr): ν = 3343, 1742, 1663,

˜

1

1524, 1233 cm^–1. H NMR (CDCl₃, 300 MHz): δ = 7.91 (s, 1 H),

6.00 (d, J = 9.9 Hz, 1 H), 5.93 (d, J = 9.1 Hz, 1 H), 5.45 (t, J =

9.9 Hz, 1 H), 5.24 (t, J = 9.8 Hz, 1 H), 4.70 (s, 2 H), 4.56 (q, J =

9.9 Hz, 1 H), 4.30 (dd, J = 12.6, 4.8 Hz, 1 H), 4.15 (dd, J = 12.6,

chloroform). IR (KBr): ν = 3514, 1748, 1729, 1236, 1100, 1039

˜

cm^–1. H NMR (CDCl₃, 300 MHz): δ = 7.79 (s, 1 H), 5.89 (m, 1 1.8 Hz, 1 H), 4.01 (ddd, J = 9.9, 4.8, 2.0 Hz, 1 H), 2.09, 2.08, 1.78

1

H), 5.48–5.39 (m, 2 H), 5.25 (m, 1 H), 4.82 (m, 2 H), 4.31 (dd, J

= 12.6, 5.0 Hz, 1 H), 4.15 (dd, J = 12.6, 2.1 Hz, 1 H), 4.01 (ddd, J

= 10.1, 5.0, 2.1 Hz, 1 H), 2.09, 2.07, 2.04, 1.89 (4 s, 12 H) ppm.

(3 s, 12 H) ppm. ¹³C NMR (CDCl₃, 75 MHz): δ = 170.8, 148.7,

122.3, 85.9, 75.0, 72.2, 68.1, 61.8, 53.5, 35.8, 22.9, 20.7, 20.7 ppm.

HRMS (FAB+): calcd. for C₁₇H₂₃ClN₄O₈[M + Na]⁺469.1102;

¹³C NMR (CDCl₃, 75 MHz): δ = 170.6, 169.9, 169.4, 169.1, 149.2, found 469.1100.

120.3, 85.8, 75.1, 72.7, 70.4, 67.8, 61.6, 56.4, 20.7, 20.6, 20.2 ppm.

1-(2-Acetamido-2-deoxy-3,4,6-tri-O-acetyl-β-D-glucopyranosyl)-4-

HRMS (FAB+): calcd. for C₁₇H₂₃N₃O₁₀[M + Na]⁺452.1281;

found 452.1286.

(azidomethyl)-1H-1,2,3-triazole (28): Obtained from 27 (446 mg)

following the general procedure for the synthesis of the azido deriv-

atives. Column chromatography (EtOAc/MeOH, 20:1) gave 28 as a

4-(Chloromethyl)-1-(2,3,4,6-tetra-O-acetyl-β-D-glucopyranosyl)-1H-

1,2,3-triazole (24): Obtained from 23 (429 mg) following the general solid (376 mg, 83%); m.p. Ͼ200 °C (dec). [α]²_D⁰= –27 (c = 1 in

1

procedure for the synthesis of chloro derivatives. Column

chromatography of the crude material (EtOAc/hexane, 2:1) gave 24

as a solid (409 mg, 91%); m.p. 166–168 °C. [α]²_D⁰= –27 (c = 1 in

chloroform). IR (KBr): ν = 2097, 1743, 1664, 1524, 1228 cm^–1. H

˜

NMR (CDCl₃, 300 MHz): δ = 7.97 (s, 1 H), 6.64 (d, J = 9.3 Hz, 1

H), 6.11 (d, J = 10.0 Hz, 1 H), 5.52 (t, J = 10.0 Hz, 1 H), 5.25 (t,

J = 9.7 Hz, 1 H), 4.63 (q, J = 10.0 Hz, 1 H), 4.54 (d, J = 14.4 Hz,

chloroform). IR (KBr): ν = 1746, 1378, 1231, 1038 cm^–1. ¹H NMR

˜

(CDCl₃, 300 MHz): δ = 7.88 (s, 1 H), 5.92 (m, 1 H), 5.49–5.41 (m, 1 H), 4.48 (d, J = 14.4 Hz, 1 H), 4.31 (dd, J = 12.6, 4.8 Hz, 1 H),

2 H), 5.27 (m, 1 H), 4.71 (s, 2 H), 4.32 (dd, J = 12.6, 5.0 Hz, 1 H), 4.16 (dd, J = 12.6, 2.9 Hz, 1 H), 4.07 (ddd, J = 10.0, 4.8, 2.1 Hz,

4.16 (dd, J = 12.6, 1.9 Hz, 1 H), 4.05 (ddd, J = 10.1, 5.0, 1.9 Hz, 1 H), 2.08, 2.07, 1.78 (3 s, 12 H) ppm. ¹³C NMR (CDCl₃, 75 MHz):

1 H), 2.09, 2.08, 2.04, 1.90 (4 s, 12 H) ppm. ¹³C NMR (CDCl₃,

75 MHz): δ = 169.8, 169.4, 168.9, 145.4, 121.2, 85.8, 75.2, 72.6,

70.3, 67.7, 61.5, 35.8, 20.6, 20.5, 20.1 ppm. HRMS (FAB+): calcd.

for C₁₇H₂₂ClN₃O₉[M + Na]⁺470.0942; found 470.0947.

δ = 170.8, 169.1, 142.9, 122.1, 86.0, 75.1, 72.3, 68.1, 61.8, 53.4,

45.3, 22.8, 20.7, 20.6 ppm. HRMS (FAB+): calcd. for C₁₇H₂₃N₇O₈

[M + Na]⁺476.1506; found 476.1508.

4-(Hydroxymethyl)-1-(2,3,4,6-tetra-O-acetyl-α-D-mannopyranosyl)-

4-(Azidomethyl)-1-(2,3,4,6-tetra-O-acetyl-β-

D

-glucopyranosyl)-1H-

1H-1,2,3-triazole (29): Obtained from 6 (373 mg) and 22 (62 mg)

following the general procedure for the CuAAC reactions using

DMF as solvent. Column chromatography (EtOAc) of the crude

material gave 29 as a syrup (395 mg, 92%); m.p. 122–124 °C. [α]²_D⁰

1,2,3-triazole (25): Obtained from 24 (447 mg) following the general

procedure for the synthesis of the azido derivatives. Column

chromatography (diethyl ether/hexane, 5:1) gave 25 as a solid

(409 mg, 90%); m.p. 81–83 °C. [α]²_D⁰= –18.2 (c = 1 in chloroform).

= 34 (c = 1 in chloroform). IR (KBr): ν = 3540, 1750, 1374, 1239

˜

1

IR (KBr): ν = 3124, 3079, 2924, 2853, 2101, 1748, 1458, 1373, 1223,

cm^–1. H NMR (CDCl₃, 300 MHz): δ = 7.76 (s, 1 H), 6.02 (d, J =

˜

1040 cm^–1. ¹H NMR (CDCl₃, 300 MHz): δ = 7.83 (s, 1 H, H-5 2.5 Hz, 1 H), 5.96 (t, J = 3.2 Hz, 1 H), 5.91 (dd, J = 8.6, 3.7 Hz, 1

triazole), 5.92 (m, 1 H, H-1), 5.49–5.41 (m, 2 H, H-2,3), 5.26 (m,

1 H, H-4), 4.51 (s, 2 H, CH₂N₃), 4.32 (dd, J = 12.7, 5.0 Hz, 1 H,

H-6), 4.16 (dd, J = 12.6, 2.1 Hz, 1 H, H-6), 4.04 (ddd, J = 10.1,

H), 5.38 (t, J = 8.8 Hz, 1 H), 4.86 (s, 2 H), 4.38 (dd, J = 12.5,

5.4 Hz, 1 H), 4.06 (dd, J = 12.5, 2.6 Hz, 1 H), 3.92 (ddd, J = 9.0,

5.4, 2.6 Hz, 1 H), 2.2, 2.2, 2–07, 2.06 (4 s, 12 H) ppm. ¹³C NMR

5.0, 2.1 Hz, 1 H, H-5), 2.09, 2.08, 2.04, 1.90 (4 s, 12 H, 4 MeCO) (CDCl₃, 75 MHz): δ = 170.5, 169.6, 169.5, 148.5, 122.4, 83.6, 71.8,

ppm. ¹³C NMR (CDCl₃, 75 MHz): δ = 169.9, 169.4, 169.0 (CO),

68.8, 68.1, 67.5, 61.5, 55.9, 20.6, 20.5, 20.4 ppm. HRMS (FAB+):

143.4 (C-4 triazole), 120.8 (C-5 triazole), 85.9 (C-1), 75.3 (C-5), calcd. for C₁₇H₂₃N₃O₁₀[M + Na]⁺452.1281; found 452.1284.

72.6, 70.3 (C-2,3), 67.7 (C-4), 61.6 (C-6), 45.5 (CH₂N₃), 20.7, 20.6,

4-(Chloromethyl)-1-(2,3,4,6-tetra-O-acetyl-α-D-mannopyranosyl)-

1H-1,2,3-triazole (30): Obtained from 29 (429 mg) following the ge-

neral procedure for the synthesis of chloro derivatives. Column

20.1 (MeCO) ppm. HRMS (FAB+): calcd. for C₁₇H₂₂N₆O₉[M +

Na]⁺477.1346; found 477.1343.

1-(2-Acetamido-2-deoxy-3,4,6-tri-O-acetyl-β-D-glucopyranosyl)-4- chromatography of the crude material (EtOAc/hexane, 2:1) gave 30

(hydroxymethyl)-1H-1,2,3-triazole (26): Obtained from 5 (372 mg) as a solid (402 mg, 90%); m.p. Ͼ 197 °C (dec). [α]²_D⁰= 41 (c = 1 in

1

and 22 (67 mg) following the general procedure for the CuAAC

reactions. Crystallization of the crude material (diethyl ether) gave

26 as a solid (407 mg, 95%); m.p. 230–232 °C. [α]²_D⁰= –26 (c = 1 in

chloroform). IR (KBr): ν = 3540, 1751, 1239, 1122, 1038 cm^–1. H

˜

NMR (CDCl₃, 300 MHz): δ = 7.80 (s, 1 H), 6.02 (d, J = 2.9 Hz, 1

H), 5.95 (t, J = 3.3 Hz, 1 H), 5.87 (dd, J = 8.7, 3.7 Hz, 1 H), 5.35

(t, J = 8.8 Hz, 1 H), 4.38 (dd, J = 12.5, 5.6 Hz, 1 H), 4.15 (d, J =

7.2 Hz, 1 H), 4.07 (d, J = 7.1 Hz, 1 H), 4.07 (dd, J = 12.5, 2.6 Hz,

1 H), 3.89 (ddd, J = 8.9, 5.0, 2.6 Hz, 1 H), 2.18, 2.09, 2.07, 2.06 (4

s, 12 H) ppm. ¹³C NMR (CDCl₃, 75 MHz): δ = 169.7, 148.4, 123.2,

83.6, 72.5, 68.7, 68.2, 66.2, 61.5, 35.8, 20.7, 20.6 ppm. HRMS

(FAB+): calcd. for C₁₇H₂₂ClN₃O₉[M + Na]⁺470.0942; found

chloroform). IR (KBr): ν = 3480, 3344, 1743, 1718, 1664, 1530,

˜

1238 cm^–1. ¹H NMR (CDCl₃, 300 MHz): δ = 7.93 (s, 1 H), 6.84 (d,

J = 9.3 Hz, 1 H), 6.09 (d, J = 9.9 Hz, 1 H), 5.49 (t, J = 9.9 Hz, 1

H), 5.26 (t, J = 9.7 Hz, 1 H), 4.78 (br. s, 2 H), 4.61 (q, J = 9.8 Hz,

1 H), 4.29 (dd, J = 12.6, 4.6 Hz, 1 H), 4.15 (dd, J = 12.6, 2.0 Hz,

1 H), 4.07 (ddd, J = 10.0, 4.7, 2.0 Hz, 1 H), 3.37 (br. s, 1 H), 2.08,

2.07, 1.73 (3 s, 12 H) ppm. ¹³C NMR (CDCl₃, 75 MHz): δ = 171.0, 470.0948.

2464

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Eur. J. Org. Chem. 2009, 2454–2473

Click Multivalent Heterogeneous Neoglycoconjugates

4-(Azidomethyl)-1-(2,3,4,6-tetra-O-acetyl-α- -mannopyranosyl)-1H- 1,2,3-triazol-1-yl]methane (34): Obtained from 1 (463 mg) and 31

D

1,2,3-triazole (31): Obtained from 30 (447 mg) following the general

procedure for the synthesis of the azido derivatives. Column

chromatography (diethyl ether/hexane, 5:1) of the crude material

gave 31 (395 mg, 87%); m.p. 117–119 °C. [α]²_D⁰= 36 (c = 1 in chloro-

(454 mg) following the general procedure for the CuAAC reactions.

Column chromatography [EtOAc/hexane (3:1)ǞEtOAc] gave 34 as

syrup (823 mg, 98%): [α]²_D⁰= 46 (c = 1 in chloroform). IR (KBr):

1

ν = 1748, 1225, 1045 cm^–1. H NMR (CDCl , 300 MHz): δ = 7.93

˜

3

form). IR (KBr): ν = 2098, 1745, 1041 cm^–1. ¹H NMR (CDCl₃,

(s, 1 H), 7.83 (s, 1 H), 6.07 (d, J = 2.6 Hz, 1 H), 5.95 (t, J = 3.1 Hz,

1 H), 5.88 (dd, J = 8.9, 3.7 Hz, 1 H), 5.72 (s, 2 H), 5.37 (t, J =

8.9 Hz, 1 H), 5.32–5.29 (m, 2 H), 5.22 (br. s, 1 H), 4.97 (d, J =

˜

300 MHz): δ = 7.82 (s, 1 H), 6.06 (d, J = 2.9 Hz, 1 H), 5.97 (t, J =

3.3 Hz, 1 H), 5.88 (dd, J = 8.7, 3.7 Hz, 1 H), 5.36 (t, J = 8.8 Hz, 1

H), 4.55 (s, 2 H), 4.39 (dd, J = 12.5, 5.6 Hz, 1 H), 4.07 (dd, J = 0.7 Hz, 1 H), 4.83 (d, J = 12.3 Hz, 1 H), 4.68 (d, J = 12.3 Hz, 1

12.5, 2.5 Hz, 1 H), 3.89 (ddd, J = 8.9, 5.6, 2.6 Hz, 1 H), 2.09, 2.08,

2.08, 2.06 (4 s, 12 H) ppm. ¹³C NMR (CDCl₃, 75 MHz): δ = 170.4,

169.6, 169.3, 143.4, 122.7, 83.5, 72.4, 68.7, 6.1, 66.1, 61.5, 45.5,

20.7, 20.6 ppm. HRMS (FAB+): calcd. for C₁₇H₂₂N₆O₉[M +

Na]⁺477.1346; found 477.1351.

H), 4.38 (dd, J = 12.5, 5.4 Hz, 1 H), 4.27 (dd, J = 12.6, 5.4 Hz, 1

H), 4.09–4.03 (m, 3 H), 3.89 (ddd, J = 8.9, 5.3, 2.5 Hz, 1 H), 2.17,

2.14, 2.11, 2.08, 2.07, 2.05, 2.04, 1.97 (8 s, 24 H) ppm. ¹³C NMR

(CDCl₃, 75 MHz): δ = 170.6, 170.4, 169.9, 169.7, 169.6, 169.5,

169.3, 142.2, 123.7, 123.5, 96.9, 83.7, 72.3, 69.3, 69.0, 68.7, 68.6,

67.9, 65.9, 65.8, 62.8, 62.3, 60.8, 45.0, 20.7, 20.6, 20.5 ppm. HRMS

(FAB+): calcd. for C₃₄H₄₄N₆O₁₉[M + Na]⁺863.2559; found

863.2560.

1-[(2,3,4,6-Tetra-O-acetyl-β-

D-glucopyranosyl)-1H-1,2,3-triazole-4-

yl]-4-[(2,3,4,6-tetra-O-acetyl-α-

D

-mannopyranosyloxymethyl)-1H-

1,2,3-triazol-1-yl]methane (32): Obtained from 1 (463 mg) and 25

(454 mg) following the general procedure for the CuAAC reactions.

Column chromatography [diethyl ether/hexane (3:1)ǞEtOAc] gave

32 as a solid (815 mg, 97%); m.p. 90 °C. [α]²_D⁰= 11 (c = 1 in chloro-

1-[(β-D-Glucopyranosyl)-1H-1,2,3-triazole-4-yl]-[4-(α-D-mannopyr-

anosyloxymethyl)-1H-1,2,3-triazol-1-yl]methane (35): Obtained

from 32 (840 mg) following the general procedure for the de-O-

acetylation (method A). Column chromatography (EtOAc/MeOH,

1:1) gave 35 as a foamy solid (464 mg, 92%): [α]²_D⁰= +33 (c = 1 in

form). IR (KBr): ν = 1746, 1224, 1039 cm^–1. ¹H NMR (CDCl₃,

˜

300 MHz): δ = 7.83 (s, 1 H, H-5 triazole), 7.65 (s, 1 H, H-5 tri-

azole), 5.78 (d, J = 8.8 Hz, 1 H, H-1 Glc), 5.64 (d, J = 15.4 Hz, 1

H, CH₂N), 5.57 (d, J = 15.4 Hz, 1 H, CH₂N), 5.38–5.26 (m, 2 H,

H-2,3 Glc), 5.21–5.15 (m, 3 H, H-3 Glc, H-3,4 Man), 5.12 (br. s, 1

H, H-2 Man), 4.85 (d, J = 1.4 Hz, 1 H, H-1 Man), 4.73 (d, J =

12.3 Hz, 1 H, CH₂O), 4.57 (d, J = 12.4 Hz, 1 H, CH₂O), 4.25–3.90

(several m, 6 H, H-5,6,6’ Glc, H-5,6,6’ Man), 2.05, 2.02, 1.98, 1.97,

1.95, 1.93, 1.87, 1.78 (8 s, 24 H, 8 Ac) ppm. ¹³C NMR (CDCl₃,

75 MHz): δ = 170.7, 170.0, 169.9, 169.3, 169.0, 168.0 (CO), 143.9,

142.1 (C-4 triazole), 123.4, 122.0 (C-5 triazole), 96.9 (C-1 Man),

86.0 (C-1 Glc), 75.3 (C-5 Glc), 72.4, 70.6 (C-2,3 Glc), 69.5, 69.0,

68.7, 67.6 (C-2,3,5 Man, C-4 Glc), 66.1 (C-4 Man), 62.4, 61.5 (C-

6 Man, C-6 Glc), 60.9 (CH₂O), 45.3 (CH₂N), 20.9, 20.8, 20.7, 20.5,

20.2 (MeCO) ppm. HRMS (FAB+): calcd. for C₃₄H₄₄N₆O₁₉[M +

Na]⁺863.2559; found 863.2563.

H O). IR (KBr): ν = 3366, 1642, 1580, 1458, 1410, 1233, 1128,

˜

2

1051 cm^–1. H NMR ([D₆]DMSO, 300 MHz): δ = 8.40 (s, 1 H, H-

1

5 triazole), 8.16 (s, 1 H, H-5 triazole), 5.70 (s, 2 H), 5.54 (d, J =

9.2 Hz, 1 H), 5.45, 5.39, 5.21 (3 br. s, 3 H), 4.72 (br. s, 1 H), 4.66

(d, J = 12.1 Hz, 1 H), 4.55 (d, J = 12.1 Hz, 1 H) ppm. ¹³C NMR

([D₆]DMSO, 75 MHz): δ = 143.8, 141.5, 124.2, 123.4, 99.0, 87.5,

80.0, 79.2, 76.8, 74.2, 72.0, 70.9, 70.1, 69.5, 67.0, 61.3, 60.7, 59.0,

49.4 ppm. HRMS (FAB+): calcd. for C₁₈H₂₈N₆O₁₁[M + Na]⁺

527.1713; found 527.1709.

1-[(2-Acetamido-2-deoxy-α-

D-mannopyranosyl)-1H-1,2,3-triazole-4-

yl]-[4-(α- -mannopyranosyloxymethyl)-1H-1,2,3-triazol-1-yl]meth-

D

ane (36): Obtained from 33 (839 mg) following the general pro-

cedure for the de-O-acetylation (method B). Column chromatog-

raphy (EtOAc/MeOH, 1:1) gave 36 as a syrup (496 mg, 91 %):

1-[(2-Acetamido-2-deoxy-3,4,6-tri-O-acetyl-α-

1H-1,2,3-triazole-4-yl]-[4-(2,3,4,6-tetra-O-acetyl-α-

D

-mannopyranosyl)-

-mannopyrano-

[α]²_D⁰= +8 (c = 1 in H₂O). [α]²⁰= +23 (c = 1, H O). IR (film): ν

˜

436

2

D

= 3455, 3327, 1658, 1539, 1134 cm^–1

.

¹H NMR ([D₆]DMSO,

syloxymethyl)-1H-1,2,3-triazol-1-yl]methane (33): Obtained from 1

(463 mg) and 28 (453 mg) following the general procedure for the

CuAAC reactions. Column chromatography (EtOAc Ǟ EtOAc/

MeOH, 15:1) gave 33 as a solid (822 mg, 98%); m.p. Ͼ 215 °C

300 MHz): δ = 8.26 (s, 1 H, H-5 triazole), 8.01 (s, 1 H, H-5 tri-

azole), 7.86 (d, J = 9.0 Hz, 1 H), 5.67 (d, J = 10.1 Hz, 1 H), 5.64

(s, 2 H), 5.25 (m, 2 H), 4.69 (s, 1 H), 4.62 (d, J = 12.1 Hz, 1 H),

4.46 (d, J = 12.1 Hz, 1 H), 4.71–4.45 (several m, 5 H), 4.01 (q, J =

9.4 Hz, 1 H), 3.70–3.23 (several m, 11 H) ppm. ¹³C NMR ([D₆]-

DMSO, 75 MHz): δ = 170.2, 144.1, 141.5, 124.4, 123.5, 99.3, 86.5,

80.2, 74.2, 73.8, 70.9, 70.3, 70.0, 67.1, 61.4, 60.8, 59.3, 54.9, 44.7,

22.8 ppm. HRMS (FAB+): calcd. for C₂₀H₃₁N₇O₁₁[M + Na]⁺

568.1979; found 568.1979.

(dec). [α]²_D⁰= +2.4 (c = 1 in chloroform). IR (KBr): ν = 1746, 1228,

˜

1043 cm^–1. ¹H NMR (CDCl₃, 300 MHz): δ = 8.06 (s, 1 H, H-5

triazole), 7.84 (s, 1 H, H-5 triazole), 6.67 (d, J = 9.2 Hz, 1 H, NH),

6.04 (d, J = 9.9 Hz, 1 H, H-1 GlcNAc), 5.71 (d, J = 15.4 Hz, 1 H,

CH₂N), 5.63 (d, J = 15.4 Hz, 1 H, CH₂N), 5.46 (t, J = 9.9 Hz, 1

H, H-3 GlcNAc), 5.26–5.18 (m, 4 H, H-4 GlcNAc, H-2,3,4 Man),

4.92 (d, J = 1.3 Hz, 1 H, H-1 Man), 4.79 (d, J = 12.3 Hz, 1 H,

CH₂O), 4.64 (d, J = 12.3 Hz, 1 H, CH₂O), 4.52 (q, J = 9.8 Hz, 1

H, H-2 GlcNAc), 4.29–4.04 (m, 6 H, H-5,6,6’ GlcNAc, H-5,6,6’

Man), 2.15, 2.11, 2.07, 2.06, 2.05, 2.03, 1.98 (7 s, 21 H, 7 Ac), 1.73

(s, 3 H, AcN) ppm. ¹³C NMR (CDCl₃, 75 MHz): δ = 170.6, 170.1,

170.0, 169.7, 169.4 (CO), 143.9, 141.6 (C-4 triazole), 123.7, 122.8

(C-5 triazole), 97.0 (C-1 Man), 86.2 (C-1 GlcNAc), 75.05 (C-5

GlcNAc), 72.1, 70.6 (C-3 GlcNAc), 69.5, 69.2, 68.7, 68.1 (C-2,3,5

Man, C-4 GlcNAc), 66.1 (C-4 Man), 62.4, 61.5 (C-6 Man, C-6

GlcNAc), 61.0 (CH₂O), 53.7 (C-2 GlcNAc), 45.3 (CH₂N), 29.7

(MeCON), 20.9, 20.8, 20.7, 20.6, 20.2 (MeCO) ppm. HRMS

(FAB+): calcd. for C₃₄H₄₅N₇O₁₈[M + Na]⁺862.2719; found

862.2713.

1-[(β-D-Glucopyranosyl)-1H-1,2,3-triazole-4-yl]-[4-(α-D-mannopyr-

anosyloxymethyl)-1H-1,2,3-triazol-1-yl]methane (37): Obtained

from 34 (840 mg) following the general procedure for the de-O-

acetylation (method B). Column chromatography (EtOAc/MeOH,

1:1) gave 37 as a foamy solid (464 mg, 92%): [α]²_D⁰= +54 (c = 1 in

1

H O). IR (KBr): ν = 3378, 1566, 1417, 1126, 1051 cm^–1. H NMR

˜

2

([D₆]DMSO + D₂O, 300 MHz): δ = 8.31 (s, 1 H, H-5 triazole), 8.12

(s, 1 H, H-5 triazole), 5.89 (d, J = 4.5 Hz, 1 H), 5.69 (s, 2 H), 4.80

(br. s, 1 H), 4.69 (s, 1 H), 4.63 (d, J = 12.2 Hz, 1 H), 4.48 (d, J =

12.2 Hz, 1 H), 4.35 (dd, J = 4.1, 3.2 Hz, 1 H), 3.81 (dd, J = 6.5,

3.1 Hz, 1 H), 3.70–3.30 (several m, 10 H) ppm. ¹³C NMR ([D₆]-

DMSO, 75 MHz): δ = 14e.8, 141.5, 124.1, 122.9, 99.0, 85.7, 78.5,

74.1, 71.1, 70.9, 70.1, 68.0, 67.7, 67.0, 61.2, 60.6, 59.0, 44.4 ppm.

HRMS (FAB+): calcd. for C₁₈H₂₈N₆O₁₁[M + Na]⁺527.1713;

found 527.1713.

1-[(2,3,4,6-Tetra-O-acetyl-β-

D-glucopyranosyl)-1H-1,2,3-triazole-4-

yl]-[4-(2,3,4,6-tetra-O-acetyl-α-

D

-mannopyranosyloxymethyl)-1H-

Eur. J. Org. Chem. 2009, 2454–2473

www.eurjoc.org

2465

F. Santoyo-Gonzalez et al.

FULL PAPER

5,5-Bis(7-chloro-2,5-dioxaheptyl)-2-(4-methoxyphenyl)-1,3-dioxane s, 2 H), 4.97 (s, 2 H), 4.84 (d, J = 12.2 Hz, 2 H), 4.69 (d, J =

(39): Obtained from 38 (254 mg) following the general procedure 12.2 Hz, 2 H), 4.55 (t, J = 5.1 Hz, 4 H), 4.31 (dd, J = 12.3, 5.1 Hz,

for the synthesis of (2-chloroethoxy)ethyl derivatives. Column 2 H), 4.14–4.07 (m, 4 H), 3.90 (t, J = 5.1 Hz, 4 H), 3.75 (t, J =

chromatography (diethyl ether/hexane, 2:1) gave 39 as a syrup

5.7 Hz, 4 H), 3.65–3.53 (m, 20 H), 3.43 (s, 8 H), 2.15, 2.12, 2.04,

1.98 (4 s, 24 H) ppm. ¹³C NMR (CDCl₃, 75 MHz): δ = 170.0,

169.9, 169.7, 143.3, 124.2, 96.8, 71.3, 71.1, 71.0, 70.5, 70.0, 69.9,

69.5, 69.4, 69.1, 68.7, 66.1, 62.4, 60.9, 45.5, 43.0, 20.9, 20.8, 20.7

ppm. HRMS (MALDI-TOF): calcd. for C₅₅H₈₄N₆O₂₈[M + Na]⁺

(382 mg, 82 %). IR (film): ν = 1614,1517,1250, 1097, 1033, 830

˜

1

cm^–1. H NMR (CDCl₃, 300 MHz): δ = 7.40 (d, J = 8.7 Hz, 2 H),

6.89 (d, J = 8.7 Hz, 2 H), 5.38 (s, 1 H), 4.09 (d, J = 11.5 Hz, 2 H),

3.87 (d, J = 11.4 Hz, 2 H), 3.82–3.53 (several m, 16 H), 3.80 (s, 4

H), 3.68 (s, 3 H, OMe) ppm. ¹³C NMR (CDCl₃, 75 MHz): δ = 1369.460; found 1369.576.

160.0, 131.0, 127.4, 113.7, 101.7, 71.4, 71.3, 71.3, 71.2, 71.0, 70.5,

1,15-Diazido-8,8-bis[7-{4-(2,3,4,6-tetra-O-acetyl-α-D-glucopyranos-

70.5, 70.0, 69.7, 55.3, 42.9, 38.9 ppm. HRMS (FAB+): calcd. for

yloxymethyl)-1H-1,2,3-triazol-1-yl}-2,5-dioxahept-1-yl]-3,6,10,13-

tetraoxapentadecane (44): Obtained from 42 (449 mg) following the

general procedure for the synthesis of the azido derivatives. Column

chromatography (EtOAc/MeOH, 20:1) gave 44 as a syrup (435 mg,

C₂₁H₃₂Cl₂O₇[M + Na]⁺489.1423; found 489.1427.

5,5-Bis(7-azido-2,5-dioxaheptyl)-2-(4-methoxyphenyl)-1,3-dioxane

(40): Obtained from 39 (466 mg) following the general procedure

for the synthesis of the azido derivatives. Column chromatography

(diethyl ether/hexane, 2:1) gave 40 as a syrup (413 mg, 86%). IR

96%): [α]²_D⁰= +61 (c = 1 in chloroform). IR (film): ν = 2109, 1751,

˜

1369, 1227, 1103, 1041 cm^–1. ¹H NMR (CDCl₃, 300 MHz): δ =

7.72 (s, 2 H), 5.47 (t, J = 9.8 Hz, 2 H), 5.21 (d, J = 3.7 Hz, 2 H),

5.08 (t, J = 9.7 Hz, 2 H), 4.88 (dd, J = 10.3, 3.8 Hz, 2 H), 4.83 (d,

J = 12.5 Hz, 2 H), 4.69 (d, J = 12.3 Hz, 2 H), 4.54 (t, J = 5.1 Hz,

4 H), 4.27 (dd, J = 12.3, 4.1 Hz, 2 H), 4.15–4.05 (m, 4 H), 3.90 (t,

J = 5.1 Hz, 4 H), 3.67 (t, J = 5.0 Hz, 4 H), 3.63–3.52 (m, 1 H),

3.42 (s, 8 H), 3.37 (t, J = 5.0 Hz, 4 H), 2.10, 2.03, 2.02, 2.00 (4 s,

24 H) ppm. ¹³C NMR (CDCl₃, 75 MHz): δ = 170.7, 170.0, 169.6,

143.7, 124.0, 95.2, 71.3, 71.1, 71.0, 70.7, 70.6, 70.5, 70.1, 70.0, 69.6,

69.5, 67.6, 61.8, 61.4, 50.8, 50.5, 45.6, 20.8, 20.7, 20.7, 20.6 ppm.

HRMS (MALDI-TOF): calcd. for C₅₅H₈₄N₁₂O₂₈[M + Na]⁺

1383.541; found 1383.806.

(film): ν = 2867, 2109, 1615, 1518, 1303, 1249, 1095, 1032, 830

˜

1

cm^–1. H NMR (CDCl₃, 300 MHz): δ = 7.40 (d, J = 8.7 Hz, 2 H),

6.89 (d, J = 8.7 Hz, 2 H), 5.38 (s, 1 H), 4.09 (d, J = 11.8 Hz, 2 H),

3.88 (d, J = 11.7 Hz, 2 H), 3.80 (s, 4 H), 3.70–3.35 (several m, 16

H), 3.67 (s, 3 H) ppm. ¹³C NMR (CDCl₃, 75 MHz): δ = 161.1,

131.0, 127.4, 113.7, 101.7, 71.4, 71.3, 71.1, 70.6, 70.6, 70.2, 70.1,

70.0, 69.8, 55.4, 50.9, 39.0 ppm. HRMS (FAB+): calcd. for

C₂₁H₃₂N₆O₇[M + Na]⁺503.2230; found 503.2230.

1,15-Dichloro-8,8-bis(7-chloro-2,5-dioxahept-1-yl)-3,6,10,13-tetra-

oxapentadecane (41): Obtained from 40 (480 mg) following the ge-

neral procedure for the acetal hydrolysis. The obtained crude mate-

rial was directly transformed without purification following the ge-

neral procedure for the synthesis of the (2-chloroethoxy)ethyl deriv-

atives. Column chromatography (diethyl ether/hexane, 1:1) gave 41

1,15-Diazido-8,8-bis[7-{4-(2,3,4,6-tetra-O-acetyl-α-D-mannopyran-

osyloxymethyl)-1H-1,2,3-triazol-1-yl}-2,5-dioxahept-1-yl]-3,6,10,13-

tetraoxapentadecane (45): Obtained from 43 (449 mg) following the

general procedure for the synthesis of the azido derivatives. Column

chromatography (EtOAc/MeOH, 20:1) gave 45 as a syrup (449 mg,

as a syrup (310 mg, 54% overall yield from 40). IR (film): ν = 2871,

˜

1

2109, 1300, 1109 cm^–1. H NMR (CDCl₃, 300 MHz): δ = 3.76 (t,

J = 6.0 Hz, 4 H), 3.70–3.54 (m, 24 H), 3.46 (s, 8 H), 3.37 (t, J =

5.0 Hz, 4 H) ppm. ¹³C NMR (CDCl₃, 75 MHz): δ = 71.3, 71.1,

70.5, 70.5, 70.1, 70.0, 50.8, 45.6, 42.9 ppm. HRMS (FAB+): calcd.

for C₂₁H₄₀Cl₂N₆O₈[M + Na]⁺597.218; found 597.218.

99%): [α]²_D⁰= +26 (c = 1 in chloroform). IR (film): ν = 2109, 1750,

˜

1

1370, 1228, 1134, 1086, 1048 cm^–1. H NMR (CDCl₃, 300 MHz):

δ = 7.77 (s, 2 H), 5.32–5.24 (m, 4 H), 5.24 (br. s, 2 H), 4.98 (d, J =

1.2 Hz, 2 H), 4.84 (d, J = 12.2 Hz, 2 H), 4.69 (d, J = 12.2 Hz, 2

H), 4.55 (t, J = 5.1 Hz, 4 H), 4.31 (dd, J = 12.5, 5.2 Hz, 2 H), 4.14–

4.10 (m, 4 H), 3.90 (t, J = 5.1 Hz, 4 H), 3.67 (t, J = 5.0 Hz, 4 H),

3.64–3.53 (m, 20 H), 3.44 (s, 8 H), 3.37 (t, J = 5.0 Hz, 4 H), 2.15,

2.12, 2.04, 1.98 (4 s, 24 H) ppm. ¹³C NMR (CDCl₃, 75 MHz): δ =

170.7, 170.0, 169.8, 169.7, 143.2, 124.2, 96.8, 71.0, 71.0, 70.5, 70.4,

70.0, 69.9, 69.4, 69.4, 69.1, 68.7, 66.0, 62.3, 60.8, 50.7, 50.4, 45.5,

20.8, 20.8, 20.7 ppm. HRMS (MALDI-TOF): calcd. for

C₅₅H₈₄N₁₂O₂₈[M + Na]⁺1383.541; found 1383.82.

1,15-Dichloro-8,8-bis[7-{4-(2,3,4,6-tetra-O-acetyl-α-D-glucopyrano-

syloxymethyl)-1H-1,2,3-triazol-1-yl}-2,5-dioxahept-1-yl]-3,6,10,13-

tetraoxapentadecane (42): Obtained from 41 (287 mg) and 2

(463 mg) following the general procedure for the CuAAC reactions.

Column chromatography (EtOAc/MeOH, 20:1) gave 42 isolated as

a syrup (606 mg, 90%): [α]²_D⁰= +60 (c = 1 in chloroform). IR (film):

ν = 1749, 1367, 1226, 1102, 1040 cm^–1¹H NMR (CDCl₃,

.

˜

300 MHz): δ = 7.73 (s, 2 H), 5.47 (t, J = 9.9 Hz, 2 H), 5.21 (d, J =

3.7 Hz, 2 H), 5.08 (t, J = 9.8 Hz, 2 H), 4.89 (dd, J = 10.2, 3.7 Hz,

2 H), 4.84 (d, J = 12.3 Hz, 2 H), 4.67 (d, J = 12.3 Hz, 2 H), 4.55

(t, J = 5.1 Hz, 4 H), 4.27 (dd, J = 12.3, 4.2 Hz, 2 H), 4.14–4.05 (m,

4 H), 3.90 (t, J = 5.1 Hz, 4 H), 3.75 (t, J = 5.8 Hz, 4 H), 3.65–3.53

(m, 1 H), 3.42 (s, 8 H), 3.37 (t, J = 5.0 Hz, 4 H), 2.10, 2.03, 2.02,

2.00 (4 s, 24 H) ppm. ¹³C NMR (CDCl₃, 75 MHz): δ = 170.1,

143.5, 124.1, 95.2, 71.3, 71.1, 71.0, 70.7, 70.5, 70.5, 70.1, 70.0, 69.6,

69.5, 68.5, 67.5, 61.8, 61.4, 50.5, 45.6, 43.0, 20.8, 20.7, 20.7 ppm.

HRMS (MALDI-TOF): calcd. for C₅₅H₈₄N₆O₂₈[M + Na]⁺

1369.460; found 1369.550.

8,8-Bis[7-{4-(2,3,4,6-tetra-O-acetyl-α-

1H-1,2,3-triazol-1-yl}-2,5-dioxahept-1-yl]-1,15-bis[4-(2,3,4,6-tetra-

O-acetyl-α- -mannopyranosyloxymethyl)-1H-1,2,3-triazol-1-yl]-

D-glucopyranosyloxymethyl)-

D

3,6,10,13-tetraoxapentadecane (46): Obtained from 44 (340 mg) and

1 (232 mg) following the general procedure for the CuAAC reac-

tions. Column chromatography (EtOAc/MeOH, 20:1) gave 46 as a

syrup (512 mg, 96%): [α]²_D⁰= +64 (c = 1 in chloroform). IR (film):

ν = 1748, 1371, 1235, 1135, 1048 cm^{–1 1}H NMR (CDCl₃,

.

˜

300 MHz): δ = 7.77, 7.73 (2 s, 4 H), 5.47 (t, J = 9.8 Hz, 2 H), 5.35–

5.25 (m, 4 H), 5.23 (s, 2 H), 5.21 (d, J = 3.9 Hz, 2 H), 5.08 (t, J =

9.8 Hz, 2 H), 4.98 (d, J = 0.7 Hz, 2 H), 4.88 (dd, J = 10.3, 3.7 Hz,

1,15-Dichloro-8,8-bis[7-{4-(2,3,4,6-tetra-O-acetyl-α-D-mannopyr-

anosyloxymethyl)-1H-1,2,3-triazol-1-yl}-2,5-dioxahept-1-yl]- 2 H), 4.83 (d, J = 12.0 Hz, 4 H), 4.68 (d, J = 12.1 Hz, 4 H), 4.54

3,6,10,13-tetraoxapentadecane (43): Obtained from 41 (287 mg) and

1 (463 mg) following the general procedure for the CuAAC reac-

tions. Column chromatography (EtOAc/MeOH, 20:1) gave 43 iso-

(t, J = 5.0 Hz, 8 H), 4.31 (dd, J = 12.3, 5.0 Hz, 2 H), 4.27 (dd, J

= 12.0, 4.2 Hz, 2 H), 4.18–4.07 (m, 8 H), 3.90 (t, J = 5.1 Hz, 8 H),

3.62–3.48 (m, 16 H), 3.37 (s, 8 H), 2.15, 2.12, 2.10, 2.04, 2.04, 2.02,

2.00, 1.98 (8 s, 48 H) ppm. ¹³C NMR (CDCl₃, 75 MHz): δ = 143.4,

143.3, 124.1, 124.0, 96.8, 95.1, 70.9, 70.6, 70.3, 70.2, 70.0, 69.8,

69.4, 69.3, 69.0, 68.7, 68.4, 67.4, 66.0, 62.3, 61.7, 61.3, 60.8, 50.3,

lated as a syrup (646 mg, 96%): [α]²_D⁰= +35 (c = 1 in chloroform).

1

IR (film): ν = 1750, 1370, 1227, 1134, 1089, 1049 cm^–1. H NMR

˜

(CDCl₃, 300 MHz): δ = 7.77 (s, 2 H), 5.35–5.25 (m, 4 H), 5.23 (br.

2466

www.eurjoc.org

Eur. J. Org. Chem. 2009, 2454–2473

Click Multivalent Heterogeneous Neoglycoconjugates

45.4, 20.8, 20.7, 20. 20.6, 20.5 ppm. HRMS (MALDI-TOF): calcd.

for C₈₉H₁₂₈N₁₂O₄₈[M + Na]⁺2155.784; found 2155.696.

azol-1-yl}-2,5-dioxahept-1-yl]-3,6,10,13-tetraoxapentadecane (50):

Obtained from 45 (340 mg) and propargyl alcohol following the

general procedure for the CuAAC reactions. Column chromatog-

raphy (CH₂Cl₂/MeOH, 12:1) gave 50 as a syrup (254 mg, 69%):

8,8-Bis[7-{4-(2,3,4,6-tetra-O-acetyl-α-

yl)-1H-1,2,3-triazol-1-yl}-2,5-dioxahept-1-yl]-1,15-bis[4-(2,3,4,6-

tetra-O-acetyl-β- -glucopyranosyloxymethyl)-1H-1,2,3-triazol-1-yl]-

D-mannopyranosyloxymeth-

[α]²_D⁰= +36 (c = 1, in chloroform). IR (film): ν = 3463, 2876, 1750,

˜

D

1

1370, 1228, 1134, 1087, 1049 cm^–1. H NMR (CDCl₃, 300 MHz):

3,6,10,13-tetraoxapentadecane (47): Obtained from 45 (340 mg) and

3 (232 mg) following the general procedure for the CuAAC reac-

δ = 7.78 (br. s, 4 H), 5.4–5.2 (m, 6 H), 4.97 (s, 4 H), 4.83 (d, J =

12.2 Hz, 2 H), 4.76 (s, 2 H), 4.75 (br. s, 2 H), 4.67 (d, J = 12.2 Hz,

2 H), 4.50 (m, 8 H), 4.30 (dd, J = 12.3, 5.1 Hz, 2 H), 4.10 (m, 4

H), 3.86 (m, 8 H), 3.57–3.48 (m, 16 H), 3.33 (s, 8 H), 2.15, 2.12,

2.04, 1.97 (4 s, 24 H) ppm. ¹³C NMR (CDCl₃, 75 MHz): δ = 170.8,

170.1, 170.0, 169.7, 143.4, 124.2, 123.0, 96.9, 71.0, 70.9, 70.5, 70.5,

69.9, 69.5, 69.4, 69.2, 68.8, 66.1, 62.4, 60.9, 56.4, 50.4, 50.3, 45.5,

20.9, 20.8, 20.7 ppm. HRMS (MALDI-TOF): calcd. for

C₆₁H₉₂O₃₀N₁₂[M + Na]⁺1495.594; found 1495.582.

tions. Column chromatography (EtOAc/MeOH, 20:1) gave 47 as a

20

syrup (426 mg, 80%): [α]²_D⁰= +3 (c = 1 in chloroform). [α]

=

436

+7.5 (c = 1, chloroform). IR (film): ν = 2877, 1751, 1433, 1370,

˜

1

1228, 1048 cm^–1. H NMR (CDCl₃, 300 MHz): δ = 7.76 (s, 2 H),

7.70 (s, 2 H), 5.32–5.29 (m, 4 H), 5.23 (br. s, 2 H), 5.20 (t, J =

9.3 Hz, 2 H), 5.09 (t, J = 9.6 Hz, 2 H), 4.93 (d, J = 12.5 Hz, 2 H),

4.84 (d, J = 12.2 Hz, 2 H), 4.81 (d, J = 12.5 Hz, 2 H), 4.71 (d, J =

8.0 Hz, 2 H), 4.68 (d, J = 12.2 Hz, 2 H), 4.56–4.50 (m, 8 H), 4.33–

4.26 (m, 4 H), 4.17–4.07 (m, 6 H), 3.88 (m, 8 H), 3.58 (m, 8 H), 8,8-Bis[7-{4-(α-

D-glucopyranosyloxymethyl)-1H-1,2,3-triazol-1-yl}-

3.50 (m, 8 H), 3.36 (s, 8 H), 2.15, 2.12, 2.09, 2.04, 2.03, 1.99, 1.97

2,5-dioxahept-1-yl]-1,15-bis[4-(α-D-mannopyranosyloxymethyl)-1H-

(8 s, 96 H) ppm. ¹³C NMR (CDCl₃, 75 MHz): δ = 170.1, 170.1, 1,2,3-triazol-1-yl]-3,6,10,13-tetraoxapentadecane (51): Obtained

169.9, 169.8, 169.6, 169.6, 169.4, 169.3 (COO), 143.8, 143.3, 124.1, from 46 (426 mg) following the general procedure for the de-O-

123.9, 99.9, 96.8, 72.8, 71.9, 71.2, 71.0, 70.4, 69.9, 69.4, 69.3, 69.1

acetylation (method B). Column chromatography (MeOH) gave 51

(C-3), 68.7, 68.4, 66.1, 62.8, 62.4, 61.9, 60.1, 50.3, 45.4, 20.8, 20.7, as a syrup (283 mg, 97%). [α]²_D⁰= +90 (c = 0.25 in water). IR (film):

20.6, 20.6, 20.5 ppm.

ν = 3408, 1053, 667 cm^–1. ¹H NMR (CD OD, 300 MHz): δ = 8.06,

˜

3

8.04 (2 s, 4 H), 4.92 (d, J = 3.6 Hz, 2 H), 4.82 (d, J = 12.4 Hz, 2

H), 4.79 (d, J = 12.4 Hz, 2 H), 4.66 (d, J = 12.4 Hz, 2 H), 4.64 (d,

J = 12.4 Hz, 2 H), 4.57 (br. t, J = 4.5 Hz, 2 H), 3.89 (t, J = 5.1 Hz,

8 H), 3.85–3.45 (m), 3.42 (dd, J = 9.7, 3.7 Hz, 2 H), 3.35–3.30 (m)

ppm. ¹³C NMR (CD₃OD, 75 MHz): δ = 145.3, 145.2, 126.1, 100.8,

99.6, 75.0, 74.9, 74.0, 73.5, 72.5, 72.1, 72.0, 71.8, 68.6, 62.9, 62.7,

61.5, 60.8, 51.5, 44.0 ppm. HRMS (MALDI-TOF): calcd. for

C₅₇H₉₆N₁₂O₃₂[M + Na]⁺1483.615; found 1483.787.

8,8-Bis[7-{4-(2,3,4,6-tetra-O-acetyl-α-D-glucopyranosyloxymethyl)-

1H-1,2,3-triazol-1-yl}-2,5-dioxahept-1-yl]-1,15-bis[4-(2,3,4,6-tetra-

O-acetyl-α- -glucopyranosyloxymethyl)-1H-1,2,3-triazol-1-yl]-

D

3,6,10,13-tetraoxapentadecane (48): Obtained from 44 (340 mg) and

2 (232 mg) following the general procedure for the CuAAC reac-

tions. Column chromatography (EtOAc/MeOH, 20:1) gave 48 as a

syrup (416 mg, 78%): [α]²_D⁰= +87 (c = 1 in chloroform). IR (film):

ν = 1751, 1369, 1228, 1137, 1021 cm^–1¹H NMR (CDCl₃,

.

˜

300 MHz): δ = 7.72 (s, 4 H), 5.46 (t, J = 9.8 Hz, 4 H), 5.21 (d, J =

3.7 Hz, 4 H), 5.08 (t, J = 9.8 Hz, 4 H), 4.88 (dd, J = 10.3, 3.7 Hz,

4 H), 4.83 (d, J = 11.8 Hz, 4 H), 4.68 (d, J = 11.8 Hz, 4 H), 4.54

(t, J = 5.0 Hz, 8 H), 4.26 (dd, J = 12.3, 4.0 Hz, 2 H), 4.13–4.05 (m,

1,15-Bis[4-(β-

8,8-bis[7-{4-(α-

D

-glucopyranosyloxymethyl)-1H-1,2,3-triazol-1-yl]-

-mannopyranosyloxymethyl)-1H-1,2,3-triazol-1-yl}-

D

2,5-dioxahept-1-yl]-3,6,10,13-tetraoxapentadecane (52): Obtained

from 47 (426 mg) following the general procedure for the de-O-

8 H), 3.90 (t, J = 5.0 Hz, 8 H), 3.61–3.48 (m, 16 H), 3.36 (s, 8 H), acetylation (method B). Column chromatography (MeOH) gave 52

2.10, 2.02, 2.00 (3 s, 48 H) ppm. ¹³C NMR (CDCl₃, 75 MHz): δ = as a syrup (233 mg, 80%). [α]²_D⁰= +12 (c = 0.5 in H₂O). IR (film):

170.7, 170.1, 169.6, 135.7, 95.2, 71.3, 71.1, 70.8, 70.5, 70.2, 70.0,

ν = 3362, 1655, 1075 cm^–1. ¹H NMR (CD OD, 300 MHz): δ = 8.00

˜

3

69.5, 68.6, 67.6, 61.9, 61.3, 50.7, 20.8, 20.7, 20.6 ppm. HRMS (s, 4 H), 4.93 (d, J = 12.4 Hz, 2 H), 4.83 (d, J = 1.0 Hz, 2 H), 4.75

(MALDI-TOF): calcd. for C₈₉H₁₂₈N₁₂O₄₈[M + Na]⁺2155.784;

found 2155.895.

(d, J = 12.4 Hz, 2 H), 4.73 (d, J = 12.4 Hz, 2 H), 4.60 (d, J =

12.4 Hz, 2 H), 4.53 (t, J = 4.8 Hz, 4 H), 4.37 (d, J = 7.7 Hz, 2 H),

3.90–3.15 (several m, 76 H) ppm. ¹³C NMR (CD₃OD, 75 MHz): δ

= 145.4, 145.0, 126.1, 126.0,103.5, 100.7, 77.9, 77.9, 74.9, 74.9,

72.4, 72.0, 71.9, 71.5, 71.3, 70.5, 70.3, 68.5, 63.0, 62.8, 62.7, 60.7,

51.5, 46.6 ppm. HRMS (MALDI-TOF): calcd. for C₅₇H₉₆N₁₂O₃₂

[M + Na]⁺1483.615; found 1483.761.

8,8-Bis[7-{4-(2,3,4,6-tetra-O-acetyl-α-

yl)-1H-1,2,3-triazol-1-yl}-2,5-dioxahept-1-yl]-1,15-bis[4-(2,3,4,6-

tetra-O-acetyl-α- -mannopyranosyloxymethyl)-1H-1,2,3-triazol-1-

D-mannopyranosyloxymeth-

D

yl]-3,6,10,13-tetraoxapentadecane (49): Obtained from 45 (340 mg)

and 1 (232 mg) following the general procedure for the CuAAC

reactions. Column chromatography (EtOAc/MeOH, 20:1) gave 49

8,8-Bis[7-{4-(α-

2,5-dioxahept-1-yl]-1,15-bis[4-(α-

1,2,3-triazol-1-yl]-3,6,10,13-tetraoxapentadecane (53): Obtained

D-glucopyranosyloxymethyl)-1H-1,2,3-triazol-1-yl}-

as a syrup (506 mg, 95%): [α]²_D⁰= +41 (c = 1 in chloroform). IR

D

-glucopyranosyloxymethyl)-1H-

1

(film): ν = 1751, 1370, 1228, 1133, 1049 cm^–1. H NMR (CDCl ,

˜

3

300 MHz): δ = 7.77 (s, 4 H, H-4 triazole), 5.35–5.24 (m, 8 H, H- from 48 (426 mg) following the general procedure for the de-O-

3,4 Man), 5.23 (s, 2 H, H-2 Man), 4.98 (s, 4 H, H-1), 4.84 (d, J =

12.2 Hz, 4 H, CH₂O-Man), 4.68 (d, J = 12.2 Hz, 4 H, CH₂O-Man),

4.55 (t, J = 5.1 Hz, 8 H, CH₂N), 4.31 (dd, J = 12.3, 5.1 Hz, 2 H,

acetylation (method B). Column chromatography (MeOH) gave 53

as a syrup (259 mg, 89%): [α]²_D⁰= +37 (c = 0.25 in H₂O). IR (film):

ν = 3407, 1655, 1460, 1230, 1044 cm^{–1 1}H NMR (CD₃OD,

.

˜

H-6), 4.15–4.06 (m, 8 H, H-5,6), 3.90 (t, J = 5.1 Hz, 8 H), 3.61– 300 MHz): δ = 8.04 (s, 4 H), 4.91 (d, J = 3.7 Hz, 4 H), 4.81 (d, J

3.51 (m, 16 H), 3.38 (s, 8 H, CH₂C), 2.15, 2.12, 2.04, 1.98 (4 s, 48

= 12.4 Hz, 4 H), 4.66 (d, J = 12.4 Hz, 4 H), 4.56 (t, J = 5.0 Hz, 8

H), 3.89 (t, J = 5.1 Hz, 4 H), 3.79 (dd, J = 11.6, 2.0 Hz, 4 H), 3.70–

H, 4 Ac) ppm. ¹³C NMR (CDCl₃, 75 MHz): δ = 170.6, 170.0,

169.9, 169.7 (COO), 143.2, 124.1 (triazole), 96.8 (C-1), 70.9, 70.4, 3.45 (m), 3.40 (dd, J = 9.8, 3.7 Hz, 4 H), 3.37–3.25 (m) ppm. ¹³C

69.8, 69.3 (CH₂O), 69.4, 69.1, 68.7, 66.0 (C-2,3,4,5), 62.3 (C-6), NMR (CD₃OD, 75 MHz): δ = 145.4, 126.0, 99.6, 75.0, 74.0, 73.5,

60.8 (CH₂O), 50.3 (CH₂N), 45.4 [(CH₂)C], 20.8, 20.7, 20.6, 20.6 71.8, 72.1, 71.4, 70.7, 70.3, 62.7, 61.4, 51.5, 46.7 ppm. HRMS

ppm. HRMS (MALDI-TOF): calcd. for C₈₉H₁₂₈N₁₂O₄₈[M +

(MALDI-TOF): calcd. for C₅₇H₉₆N₁₂O₃₂[M + Na]⁺1483.615;

found 1483.766.

Na]⁺2155.784; found 2155.048.

1,15-Bis[4-hydroxymethyl-1H-1,2,3-triazol-1-yl]-8,8-bis[7-{4-

8,8-Bis[7-{4-(α-

D

-mannopyranosyloxymethyl)-1H-1,2,3-triazol-1-

(2,3,4,6-tetra-O-acetyl-α-D-mannopyranosyloxymethyl)-1H-1,2,3-tri- yl}-2,5-dioxahept-1-yl]-1,15-bis[4-(α-D-mannopyranosyloxymethyl)-

Eur. J. Org. Chem. 2009, 2454–2473

www.eurjoc.org

2467

F. Santoyo-Gonzalez et al.

FULL PAPER

1H-1,2,3-triazol-1-yl]-3,6,10,13-tetraoxapentadecane (54): Obtained

from 49 (426 mg) following the general procedure for the de-O-

acetylation (method B). Column chromatography (MeOH) gave 54

as a syrup (271 mg, 93%): [α]²_D⁰= +37 (c = 0.25 in H₂O). ¹H NMR

(CD₃OD, 300 MHz): δ = 8.04 (s, 4 H), 4.87 (s, 4 H), 4.79 (d, J =

12.4 Hz, 4 H), 4.64 (d, J = 12.4 Hz, 4 H), 4.57 (t, J = 5.0 Hz, 8 H),

3.89 (t, J = 5.0 Hz, 8 H), 3.85 (dd, J = 12.0, 2.3 Hz, 4 H), 3.82–

3.45 (several m, 36 H) 3.31 (s, 8 H) ppm. ¹³C NMR (CD₃OD,

75 MHz): δ = 145.3, 126.1, 100.8, 74.9, 72.5, 72.0, 72.1, 71.4, 70.7,

70.3, 68.6, 62.9, 60.8, 51.5, 46.7 ppm. HRMS (MALDI-TOF):

calcd. for C₅₇H₉₆O₃₂N₁₂[M + Na]⁺1483.615; found 1483.636.

m, 5 H), 5.54 (s, 1 H), 4.88 (d, J = 3.7 Hz, 1 H), 4.50–4.45 (m, 3

H), 4.39 (dd, J = 15.4, 2.3 Hz, 1 H), 4.29 (dd, J = 9.6, 4.2 Hz, 1

H), 3.99 (t, J = 9.2 Hz, 1 H), 3.83 (dt, J = 10.0, 4.4 Hz, 1 H), 3.74

(t, J = 9.8 Hz, 1 H), 3.70 (dd, J = 3.7, 9.2 Hz, 1 H), 3.60 (t, J =

9.2 Hz, 1 H), 3.44 (s, 3 H), 2.46 (t, J = 2.34 Hz, 1 H), 2.43 (t, J =

9.2 Hz, 1 H) ppm. ¹³C NMR (CDCl₃, 75 MHz): δ = 129.04, 128.28,

126.13, 101.42, 99.35, 82.07, 77.99, 69.09, 62.18, 60.08, 59.45, 55.34

ppm. HRMS (FAB+): calcd. for C₂₀H₂₂O₆[M + Na]⁺381.1314;

found 381.1314.

Methyl 2,3-Di-O-propargyl-α-D-glucopyranoside (61): Obtained

from 60 (358 mg) following the general procedure for the acetal

hydrolysis. Column chromatography (EtOAc) gave 61 (254 mg,

1,15-Bis[4-hydroxymethyl-1H-1,2,3-triazol-1-yl]-8,8-bis[7-{4-(α-D-

1

94%) as a syrup: [α]²_D⁰= +90 (c = 1 in MeOH). H NMR (CDCl₃,

mannopyranosyloxymethyl)-1H-1,2,3-triazol-1-yl}-2,5-dioxahept-1-

yl]-3,6,10,13-tetraoxapentadecane (55): Obtained from 50 (294 mg)

following the general procedure for the de-O-acetylation (method

B). Column chromatography (MeOH) gave 55 as a syrup (92%):

300 MHz): δ = 4.92 (d, J = 3.5 Hz, 1 H), 4.52 (dd, J = 15.8, 2.4 Hz,

1 H), 4.41 (dd, J = 15.8, 2.3 Hz, 1 H), 4.34 (m, 2 H), 3.84–3.54

(several m, 8 H), 3.44 (s, 3 H), 2.53 (t, J = 2.3 Hz, 1 H), 2.50 (t, J

= 2.3 Hz, 1 H) ppm. ¹³C NMR (CDCl₃, 75 MHz): δ = 98.04, 80.79,

80.55, 79.69, 79.26, 75.17, 74.95, 70.97, 69.88, 62.11, 60.37, 58.45,

55.25 ppm. HRMS (FAB+): calcd. for C₁₃H₁₈O₆[M + Na]⁺

293.1001; found 293.1000.

[α]²_D⁰= +29 (c = 0. 5 in H₂O). H NMR (CD₃OD, 300 MHz): δ =

1

8.00 (s, 2 H), 7.90 (s, 2 H), 4.81 (s, 2 H), 4.75 (d, J = 12.4 Hz, 2

H), 4.63 (s, 4 H), 4.60 (d, J = 12.4 Hz, 2 H), 4.53 (m, 8 H), 3.90–

3.40 (m, 36 H), 3.27 (s, 8 H) ppm. ¹³C NMR (CD₃OD, 75 MHz):

δ = 148.9, 145.1, 125.9, 124.7, 100.8, 74.9, 72.5, 72.0, 71.4, 70.7,

70.4, 70.3, 68.6, 62.9, 60.7, 56.5, 51.5, 51.4, 46.7 ppm. HRMS

(MALDI-TOF): calcd. for C₄₅H₇₆O₂₂N₁₂[M + Na]⁺1159.509;

found 1159.522.

Methyl 4,6-Di-O-chloroethoxyethyl-2,3-di-O-propargyl-α-D-gluco-

pyranoside (62): Obtained from 61 (270 mg) following the general

procedure for the synthesis of the (2-chloroethoxy)ethyl derivatives.

Column chromatography (EtOAc/hexane, 1:1) gave 62 as a syrup

(260 mg, 54%): [α]²_D⁰= +54 (c = 1 in chloroform). ¹H NMR

(CDCl₃, 300 MHz): δ = 4.87 (d, J = 3.6 Hz, 1 H), 4.44 (d, J =

2.4 Hz, 2 H), 4.35 (d, J = 2.3 Hz, 1 H), 4.00 (ddd, J = 10.8, 5.0,

3.7 Hz, 1 H), 3.85–3.56 (m, 20 H), 3.42 (dd, J = 10.0, 9.0 Hz, 1 H),

3.38 (s, 3 H), 2.43 (t, J = 2.3 Hz, 1 H), 2.42 (t, J = 2.3 Hz, 1 H)

ppm. ¹³C NMR (CDCl₃, 75 MHz): δ = 98.0, 81.3, 79.0, 77.9, 70.1,

79.9, 74.9, 74.0, 72.1, 71.3, 71.2, 70.9, 70.8, 70.5, 69.5, 60.3, 58.7,

55.0, 42.8 ppm. HRMS (FAB+): calcd. for C₂₁H₃₂O₈[M + Na]⁺

505.1372; found 505.1377.

2-(4-Methoxyphenyl)-5,5-bis(prop-2-ynyloxymethyl)-1,3-dioxane

(56): Obtained from 38 (254 mg) following the general procedure

for the propargylation. Column chromatography (diethyl ether/hex-

ane, 1:1) gave 56 isolated as a syrup (307 mg, 93%). IR (KBr): ν =

˜

1

3291, 2116, 1615, 1517, 830 cm^–1. H NMR (CDCl₃, 300 MHz): δ

= 7.41 (d, J = 8.7 Hz, 2 H), 6.89 (d, J = 8.7 Hz, 2 H), 5.38 (s, 1

H), 4.20 (d, J = 2.3 Hz, 2 H), 4.11 (d, J = 2.1 Hz, 2 H), 4.09 (d, J

= 2.4 Hz, 2 H), 3.87 (d, J = 11.4 Hz, 2 H), 3.86 (s, 2 H), 3.79 (s, 3

H), 3.36 (s, 2 H), 2.43 (m, 2 H) ppm. ¹³C NMR (CDCl₃, 75 MHz):

δ = 160.2, 131.4, 127.4, 113.7, 101.8, 74.6, 74.3, 69.9, 69.9, 68.8,

58.8, 55.4, 38.6 ppm. HMRS (FAB+): calcd. for C₁₉H₂₂O₅[M +

Na]⁺353.1365; found 353.1359.

Methyl 3,4-Di-O-isopropylidene-2,6-di-O-propargyl-α-D-galactopyr-

anoside (64): Obtained from 63 (234 mg) following the general pro-

cedure for the propargylation. Column chromatography (diethyl

ether/hexane, 1:1) gave 64 isolated as a syrup (273 mg, 88%): [α]²_D⁰

2,2-Bis(prop-2-ynyloxymethyl)-1,3-propanediol (57): Obtained from

56 (330 mg) following the general procedure for the acetal hydroly-

sis. Column chromatography (EtOAc/hexane, 1:1) gave 57 as a

= +62 (c = 1 in chloroform). IR (KBr): ν = 3265, 2117, 1242, 1221,

˜

1

1101, 1073, 1042 cm^–1. H NMR (CDCl₃, 300 MHz): δ = 4.87 (d,

syrup (199 mg, 94%). IR (KBr): ν = 3291, 2116, 1615, 1517, 830

˜

J = 3.5 Hz, 1 H), 4.42 (m, 4 H), 4.33–4.14, 3.87–3.75 (2 m, 6 H),

3.44 (s, 3 H), 2.47 (t, J = 2.4 Hz, 1 H), 2.45 (t, J = 2.3 Hz, 1 H),

1.56, 1.35 (2 s, 6 H) ppm. ¹³C NMR (CDCl₃, 75 MHz): δ = 109.4,

98.3, 79.8, 79.6, 75.9, 75.9, 73.9, 66.5, 74.9, 74.7, 69.4, 58.7, 57.9,

55.5, 28.2, 26.4 ppm. HRMS (FAB+): calcd. for C₁₆H₂₂O₆[M +

Na]⁺333.1314; found 333.1317.

1

cm^–1. H NMR (CDCl₃, 300 MHz): δ = 7.41 (d, J = 8.7 Hz, 2 H),

6.89 (d, J = 8.7 Hz, 2 H), 5.38 (s, 1 H), 4.20 (d, J = 2.3 Hz, 2 H),

4.11 (d, J = 2.1 Hz, 2 H), 4.09 (d, J = 2.4 Hz, 2 H), 3.87 (d, J =

11.4 Hz, 2 H), 3.86 (s, 2 H), 3.79 (s, 3 H), 3.36 (s, 2 H), 2.43 (m, 2

H) ppm. ¹³C NMR (CDCl₃, 75 MHz): δ = 79.5, 74.8, 70.9, 64.1,

58.8, 45.0 ppm. HMRS (FAB+): calcd. for C₁₁H₁₆O₄[M + Na]⁺

235.0944; found 235.0944.

Methyl 2,6-Di-O-propargyl-α-D-galactopyranoside (65): Obtained

from 64 (310 mg) following the general procedure for the acetal

hydrolysis Column chromatography (EtOAc) gave 65 as a syrup

1,15-Dichloro-8,8-bis(prop-2-ynyloxymethyl)-3,6,10,13-tetraoxapen-

tadecane (58): Obtained from 57 (212 mg) following the general

procedure for the synthesis of the (2-chloroethoxy)ethyl derivatives.

Column chromatography (EtOAc/hexane, 1:1) gave 58 as a syrup

(257 mg, 95%): [α]²_D⁰= +101 (c = 1 in MeOH). IR (film): ν = 3450,

˜

1

3261, 2216, 1143, 1093, 1044 cm^–1. H NMR (CDCl₃, 300 MHz):

δ = 5.02 (d, J = 3.3 Hz, 1 H, H-1), 4.41 (dd, J = 16.2, 2.4 Hz, 1 H,

OCH₂CϵCH), 4.34 (dd, J = 16.2, 2.3 Hz, 1 H, OCH₂CϵCH), 4.23

(dd, J = 16.0, 2.4 Hz, 1 H, OCH₂CϵCH), 4.17 (dd, J = 16.0,

2.4 Hz, 1 H, OCH₂CϵCH), 4.10 (dd, J = 3.2, 1.0 Hz, 1 H, H-4),

4.02–3.96 (m, 2 H, H-3,5), 3.92 (dd, J = 9.8, 3.3 Hz, 1 H, H-2),

3.81 (dd, J = 10.0, 4.8 Hz, 1 H, H-6), 3.81 (dd, J = 10.0, 6.0 Hz, 1

H, H-6’), 3.46 (s, 3 H, OMe), 2.90–2.70 (br. s, 2 H, OH), 2.52 (t, J

(271 mg, 64%). IR (film): ν = 3292, 2115, 1096 cm^–1. ¹H NMR

˜

(CDCl₃, 300 MHz): δ = 4.12 (d, J = 2.3 Hz, 4 H), 3.76 (t, J =

5.8 Hz, 4 H), 3.69–3.57 (m, 12 H), 3.52 (s, 4 H), 3.46 (s, 4 H), 2.41

(t, J = 2.3 Hz, 2 H) ppm. ¹³C NMR (CDCl₃, 75 MHz): δ = 80.2,

71.4, 71.1, 70.5, 69.9, 69.2, 58.7, 45.2, 42.9 ppm. HMRS (FAB+):

calcd. for C₁₉H₃₀O₆Cl₂[M + Na]⁺447.131; found 447.131.

Methyl 4,6-O-Benzylidene-2,3-di-O-propargyl-α-

D

-glucopyranoside

= 2.4 Hz, 1 H, CϵCH), 2.50 (t, J = 2.3 Hz, 1 H, CϵCH) ppm. ¹³

C

(60): Obtained from 59 (283 mg) following the general procedure

for the propargylation. Column chromatography (diethyl ether/hex-

ane, 1:1) gave 60 isolated as a syrup (351 mg, 98%): [α]²_D⁰= +33 (c

= 1 in chloroform). ¹H NMR (CDCl₃, 300 MHz): δ = 7.47, 7.37 (2

NMR (CDCl₃, 75 MHz): δ = 97.8 (C-1), 79.9, 79.4 (CϵCH), 76.2

(C-2), 75.4, 75.1 (CϵCH), 69.9 (C-4), 69.6 (C-6), 69.3 (C-3), 68.3

(C-5), 58.9, 58.2 (CH₂CϵCH), 55.4 (MeO) ppm. HMRS (FAB+):

calcd. for C₁₃H₁₈O₆[M + Na]⁺293.100; found 293.100.

2468

www.eurjoc.org

Eur. J. Org. Chem. 2009, 2454–2473

Click Multivalent Heterogeneous Neoglycoconjugates

1

Methyl 3,4-Di-O-chloroethoxyethyl-2,6-di-O-propargyl-α-

D-galacto-

1226, 1138, 1091, 1048, 980 cm^–1. H NMR (CDCl₃, 300 MHz): δ

pyranoside (66): Obtained from 65 (270 mg) following the general

procedure for the synthesis of the (2-chloroethoxy)ethyl derivatives.

Column chromatography (EtOAc/hexane, 1:1) gave 66 as a syrup

= 7.70 (s, 2 H), 5.25–5.20 (m, 6 H), 4.81 (s, 2 H), 4.61 (br. s, 8 H),

4.22 (dd, J = 12.0, 5.1 Hz, 2 H), 4.18–4.10 (m, 2 H), 4.04 (br. d, J

= 12.1 Hz, 2 H), 3.94–3.87 (m, 2 H), 3.75 (t, J = 5.6 Hz, 4 H),

3.65–3.54 (m, 14 H), 3.51 (s, 4 H), 3.45 (s, 4 H), 2.14, 2.10, 2.05,

(265 mg, 55%): [α]²_D⁰= +61 (c = 1 in chloroform). IR (film): ν =

˜

3288, 3255, 2215, 1352, 1096, 1049 cm^–1

.

¹H NMR (CDCl₃, 1.99 (s, 24 H) ppm. ¹³C NMR (CDCl₃, 75 MHz): δ = 170.5, 169.9,

300 MHz): δ = 4.90 (d, J = 3.7 Hz, 1 H, H-1), 4.41 (dd, J = 16.0,

2.4 Hz, 1 H), 4.35 (dd, J = 16.0, 2.3 Hz, 1 H), 4.25 (dd, J = 15.8,

169.8, 169.7, 145.6, 123.7, 97.5, 71.2, 71.0, 70.4, 69.7, 69.3, 69.9,

69.9, 69.2, 65.7, 66.3, 64.8, 62.2, 49.6, 45.4, 43.0, 20.9, 20.8, 20.8,

2.4 Hz, 1 H), 4.28 (dd, J = 15.8, 2.3 Hz, 1 H), 4.12–3.60 (m, 22 H), 20.7 ppm. HMRS (MALDI-TOF): calcd. for C₅₁H₇₆N₆O₂₆Cl₂[M

3.41 (s, 3 H), 2.47 (t, J = 2.3 Hz, 1 H), 2.43 (t, J = 2.3 Hz, 1 H)

ppm. ¹³C NMR (CDCl₃, 75 MHz): δ = 98.8, 79.6, 76.1, 75.6, 74.7,

74.4, 72.5, 71.2, 71.2, 71.0, 70.8, 70.2, 69.1, 68.6, 59.0, 58.6, 55.3,

42.9, 42.9 ppm. HRMS (FAB+): calcd. for C₂₁H₃₂O₈Cl₂[M +

Na]⁺505.1372; found 505.1369.

+ Na]⁺1281.408; found 1281.393.

1,15-Diazido-8,8-bis[1-(2,3,4,6-tetra-O-α-D-mannopyranosyloxy-

ethyl)-1H-1,2,3-triazol-1-yl-4-methyloxy]-3,6,10,13-tetraoxapenta-

decane (72): Obtained from 71 (419 mg) following the general pro-

cedure for the synthesis of azido derivatives. Column chromatog-

raphy (EtOAc/MeOH, 15:1) gave 72 as a syrup (411 mg, 97%):

4,6:4Ј,6Ј-Di-O-benzylidene-2,3,2Ј,3Ј-tetra-O-propargyl-α,α-trehalose

(68): Obtained from 67 (518 mg) following the general procedure

for the propargylation Column chromatography (diethyl ether/hex-

ane, 1:1) gave 68 isolated as a syrup (516 mg, 77%): [α]²_D⁰= +60 (c

[α]²_D⁰= +22 (c = 1 in chloroform). IR (film): ν = 2110, 1748, 1370,

˜

1226, 1138, 1091, 1048, 980 cm^–1. H NMR (CDCl₃, 300 MHz): δ

1

= 7.69 (s, 2 H, H-triazole), 5.25–5.20 (m, 6 H, H-2,3,4), 4.81 (s, 2

H, H-1), 4.60 (br. s, 8 H, OCH₂CH₂-triazole, CH₂-triazole), 4.22

(dd, J = 12.0, 5.1 Hz, 2 H, H-6), 4.18–4.11 (m, 2 H, 2 CH₂O-sugar),

4.04 (dd, J = 12.4, 2.3 Hz, 2 H, H-6’), 3.91 (m, 2 H, 2 CH₂O-

sugar), 3.67 (t, J = 5.0 Hz, 4 H, N₃CH₂CH₂O), 3.63–3.54 (m, 10

H, 2 N₃CH₂CH₂OCH₂CH₂, H-5), 3.51 (s, 4 H, OCH₂C), 3.46 (s,

4 H, OCH₂C), 3.37 (t, J = 5.0 Hz, 4 H, N₃CH₂), 2.14, 2.10, 2.04,

1.99 (4 s, 24 H, 4 Ac) ppm. ¹³C NMR (CDCl₃, 75 MHz): δ = 170.5,

169.9, 169.8, 169.6 (CO), 145.7 (C=CH), 123.6 (C=CH), 97.6 (C-

1), 71.0, 70.4, 70.0, 69.8, 69.5 (CH₂OCH₂CH₂OCH₂CCH₂), 69.2,

69.0, 68.9, 65.7 (C-2,3,4,5), 66.3 (CH₂O-sugar), 64.8 (CH₂C=), 62.2

(C-6), 50.8 (CH₂N₃), 49.5 (CH₂N), 45.5 (C_quat), 20.8, 20.7, 20.7,

20.7 (MeCO) ppm. HMRS (MALDI-TOF): calcd. for

C₅₁H₇₆N₁₂O₂₆[M + Na]⁺1295.489; found 1295.452.

= 1 in chloroform). IR (film): ν = 3287, 2250, 2118, 1453, 1372,

˜

1

1088, 989, 752 cm^–1. H NMR (CDCl₃, 300 MHz): δ = 7.55–7.30

(m, 10 H), 5.56 (s, 2 H), 5.26 (d, J = 3.9 Hz, 2 H), 4.55 (dd, J =

15.3, 2.4 Hz, 2 H), 4.46 (dd, J = 16.0, 2.4 Hz, 2 H), 4.44 (dd, J =

14.88, 2.4 Hz, 2 H), 4.39 (dd, J = 15.8, 2.4 Hz, 2 H), 4.34 (dd, J =

9.7, 4.9 Hz, 2 H), 4.24 (dt, J = 9.8, 4.9 Hz, 2 H), 3.99 (t, J = 9.2 Hz,

2 H), 4.75–3.67 (m, 4 H), 3.63 (t, J = 9.5 Hz, 2 H), 2.48 (t, J =

2.4 Hz, 2 H), 2.46 (t, J = 2.4 Hz, 2 H) ppm. ¹³C NMR (CDCl₃,

75 MHz): δ = 137.4, 129.0, 128.3, 126.1, 101.3, 94.9, 82.0, 80.2,

80.0, 78.2, 78.1, 74.8, 74.3, 69.0, 62.6, 60.2, 59.4 ppm. HRMS

(FAB+): calcd. for C₃₈H₃₈O₁₁[M + Na]⁺693.2312; found

693.2310.

2,3,2Ј,3Ј-Tetra-O-propargyl-α,α-trehalose (69): Obtained from 68

(670 mg) following the general procedure for the acetal hydrolysis

Column chromatography (EtOAc) gave 69 as a syrup (440 mg,

8,8-Bis[1-(2,3,4,6-tetra-O-α-

oxy-1H-1,2,3-triazole-1,4-diyl]-1,15-bis[4-(2,3,4,6-tetra-O-α-

D

-mannopyranosyloxyethyl)-4-methyl-

-gluco-

D

pyranosyloxymethyl)-1H-1,2,3-triazol-1-yl]-3,6,10,13-tetraoxapenta-

decane (73): Obtained from 2 (231 mg) and 72 (318 mg) following

the general procedure for the CuAAC reactions. Column

chromatography (EtOAc/MeOH, 15:1) gave 73 as a syrup (424 mg,

89%): [α]²_D⁰= +125 (c = 1 in MeOH). IR (film): ν = 3451, 3284,

˜

1

2930, 2116, 1093, 1057, 998 cm^–1. H NMR (CDCl₃, 300 MHz): δ

= 5.27 (d, J = 3.7 Hz, 2 H), 4.55 (dd, J = 15.8, 2.4 Hz, 2 H), 4.42

(dd, J = 15.9, 2.3 Hz, 2 H), 4.29 (d, J = 2.3 Hz, 2 H), 4.03 (dt, J

= 9.4, 3.7 Hz, 2 H), 3.89 (dd, J = 11.9, 3.1 Hz, 2 H), 3.82 (dd, J =

11.9, 4.4 Hz, 2 H), 3.78 (t, J = 9.2 Hz, 2 H), 3.65–3.54 (m, 4 H),

2.08 (br. s, 4 H), 2.54 (t, J = 2.3 Hz, 2 H), 2.48 (t, J = 2.3 Hz, 2

H) ppm. ¹³C NMR (CDCl₃, 75 MHz): δ = 94.0, 80.7, 80.5, 79.7,

79.0, 75.2, 75.0, 71.6, 69.9, 62.3, 60.4, 58.4 ppm. HRMS (FAB+):

calcd. for C₂₄H₃₀O₁₁[M + Na]⁺517.1686; found 517.1686.

83%): [α]²_D⁰= +57.5 (c = 1 in chloroform). IR (KBr): ν = 1751,

˜

1228, 1339, 1089, 1047 cm^–1. ¹H NMR (CDCl₃, 300 MHz): δ =

7.71, 7.69 (2 s, 2 H, H-5 triazole), 5.47 (t, 2 H, H-3 Glc), 5.29–5.18

(m, 6 H, H-2,3,4 Man), 5.22 (d, J = 3.6 Hz, 2 H, H-1 Glc), 5.08 (t,

J = 9.9 Hz, 2 H, H-4 Glc), 4.88 (dd, J = 10.3, 3.6 Hz, 2 H, H-2

Glc), 4.82 (d, J = 12 Hz, 2 H, Glc-OCH₂), 4.81 (br. s, 2 H, H-1

Man), 4.67 (d, J = 12 Hz, 2 H, Glc-OCH₂), 4.60 (t, J = 5.0 Hz, 4

H, CH₂N), 4.58 (br. s, 4 H, OCH₂-triazole), 4.53 (t, J = 5.0 Hz, 4

H, CH₂N), 4.27 (dd, J = 12.1, 4.1 Hz, 2 H, H-6 Glc), 4.22 (dd, J

= 12.2, 5.0 Hz, 2 H, H-6 Man), 4.17–4.03 (m, 8 H, H-5,6 Glc, H-

6 Man, OCH₂-Man), 3.92 (m, 2 H, OCH₂-Man), 3.88 (t, J =

4.9 Hz, 4 H, OCH₂CH₂N), 3.63 (m, 2 H, H-5 Man), 3.60–3.48 (m,

8 H, OCH₂CH₂O), 3.48, 3.40 [2 s, 8 H, C(CH₂)₄], 2.14, 2.10, 2.10,

2.04, 2.02, 2.02, 1.9 (8 s, 48 H, 16 Ac) ppm. ¹³C NMR (CDCl₃,

75 MHz): δ = 170.7, 170.6, 170.1, 170.0, 169.6 (CO), 145.4, 143.4

(C-4 triazole), 124.1, 123.8 (C-5 triazole), 97.6 (C-1 Man), 95.1 (C-

1 Glc), 71.1 (OCH₂CH₂O), 70.7 (C-2 Glc), 70.5 (OCH₂CH₂O), 70.2

(C-3 Glc), 69.9 (CCH₂O), 69.5 (OCH₂CH₂N), 69.4 (CCH₂O), 69.2

(C-2 Man), 69.1, 69.0 (C-3,5 Man), 68.6 (C-4 Glc), 67.6 (C-5 Glc),

66.3 (OCH₂CH₂N), 65.8 (C-4 Man), 64.8 (OCH₂-triazole), 62.3 (C-

6 Man), 61.8 (C-6 Glc), 61.4 (CH₂O-Man), 50.5, 49.7 (2 CH₂N),

45.5 [C(CH₂)₄], 20.9, 20.8, 20.7, 20.6 (8 MeCO) ppm. HMRS

(MALDI-TOF): calcd. for C₈₅H₁₂₀N₁₂O₄₆[M + Na]⁺2067.731;

found 2067.771.

4,6:4Ј,6Ј-Tetra-O-chloroethoxyethyl-2,3,2Ј,3Ј-tetra-O-propargyl-α,α-

trehalose (70): Obtained from 69 (494 mg) following the general

procedure for the synthesis of the (2-chloroethoxy)ethyl derivatives.

Column chromatography (diethyl ether) gave 70 as a syrup

(598 mg, 65%): [α]²_D⁰= +80 (c = 1 in chloroform). IR (film): ν =

˜

3291, 2872, 2117, 1454, 1356, 1092 cm^–1

.

¹H NMR (CDCl₃,

300 MHz): δ = 5.18 (d, J = 3.6 Hz, 2 H), 4.47 (d, J = 2.1 Hz, 4 H),

4.26 (m, 4 H), 4.08–3.98 (m, 4 H), 3.85–3.50 (m, 40 H), 3.43 (t, J

= 9.6 Hz, 2 H), 2.48 (t, J = 2.3 Hz, 2 H), 2.42 (t, J = 2.3 Hz, 2 H)

ppm. ¹³C NMR (CDCl₃, 75 MHz): δ = 94.0, 81.1, 80.4, 79.8, 78.6,

77.9, 74.7, 74.0, 72.0, 71.3, 71.1, 70.8, 70.5, 70.4, 69.4, 60.4, 58.2,

42.8, 42.7 ppm. HRMS (MALDI-TOF): calcd. for C₄₀H₅₈Cl₄O₁₅

[M + Na]⁺943.258; found 943.258.

1,15-Dichloro-8,8-bis[1-(2,3,4,6-tetra-O-α-D-mannopyranosyloxy-

ethyl)-1H-1,2,3-triazol-1-yl-4-methyloxy]-3,6,10,13-tetraoxapenta-

decane (71): Obtained from 58 (212 mg) and 7 (500 mg) following

the general procedure for the CuAAC reactions. Column

chromatography (EtOAc/MeOH, 15:1) gave 71 as a syrup (604 mg,

8,8-Bis[1-(2,3,4,6-tetra-O-α-D-mannopyranosyloxyethyl)-4-methyl-

oxy-1H-1,2,3-triazole-1,4-diyl]-1,15-bis[4-(2,3,4,6-tetra-O-β-

96%): [α]²_D⁰= +21 (c = 1 in chloroform). IR (film): ν = 1748, 1370,

D-gluco-

˜

Eur. J. Org. Chem. 2009, 2454–2473

www.eurjoc.org

2469

F. Santoyo-Gonzalez et al.

FULL PAPER

pyranosyloxymethyl)-1H-1,2,3-triazol-1-yl]-3,6,10,13-tetraoxapenta-

(4 s, 24 H, 4 Ac) ppm. ¹³C NMR (CDCl₃, 75 MHz): δ = 170.5,

decane (74): Obtained from 3 (231 mg) and 72 (218 mg) following 169.8, 169.7, 169.6 (COO), 145.4, 144.9, 124.4, 124.3 (triazole),

the general procedure for the CuAAC reactions. Column

97.6, 97.5 (C-1), 81.4, 79.5, 78.0, 72.1, 71.2, 71.1, 70.8, 70.7, 70.5,

70.0, 69.5, 69.1, 69.1, 68.9, 66.3, 66.2, 65.7, 64.2, 64.2, 55.0, 49.5,

43.0, 42.8, 20.7, 20.6, 20.6 (MeCO) ppm. HRMS (FAB+): calcd.

chromatography (EtOAc/MeOH, 15:1) gave 74 as a syrup (445 mg,

20

87%): [α]²_D⁰= –4.2 (c = 1 in chloroform). [α] = –9.3 (c = 1, chloro-

436

form). IR (KBr): ν = 1752, 1229, 1089, 1048 cm^–^{1 1}H NMR for C₅₃H₇₈ClO₂₈N₆[M + Na]⁺1339.414; found 1339.336.

.

˜

(CDCl₃, 300 MHz): δ = 7.71, 7.72 (2 s, 2 H), 5.28–5.17 (m, 8 H),

5.09 (t, J = 9.5 Hz, 2 H), 4.97 (br. t, J = 8.0 Hz, 2 H), 4.92 (d, J =

12.6 Hz, 2 H), 4.82 (br. s, 2 H), 4.79 (d, J = 12.6 Hz, 2 H), 4.71 (d,

J = 7.9 Hz, 2 H), 4.64–4.50 (m, 12 H), 4.28 (dd, J = 12.4, 4.6 Hz,

2 H), 4.22 (dd, J = 12.5, 5.1 Hz, 2 H), 4.15 (dd, J = 10.3, 2.2 Hz,

2 H), 4.15–4.10 (m, 2 H), 4.05 (dd, J = 12.4, 2.3 Hz, 2 H), 3.95–

3.88 (m, 2 H), 3.86 (t, J = 5.2 Hz, 4 H), 3.76 (ddd, J = 9.9, 4.4,

2.3 Hz, 2 H), 3.64 (m, 2 H), 3.58–3.47 (m, 8 H), 3.46, 3.39 (2 s, 8

H), 2.14, 2.09, 2.04, 2.03, 1.99, 1.98, 1.97 (7 s, 48 H) ppm. ¹³C

NMR (CDCl₃, 75 MHz): δ = 170.5, 170.1, 169.9, 169.5, 145.3,

143.7, 124.1, 123.8, 99.7, 97.5, 72.8, 71.8, 71.2, 70.9, 70.3, 69.8,

69.4, 69.1, 68.9, 68.9, 68.3, 66.2, 65.7, 64.8, 62.7, 62.2, 61.8, 50.4,

49.6, 45.3, 20.8, 20.7, 20.6, 20.6 ppm. HMRS (MALDI-TOF):

calcd. for C₈₅H₁₂₀N₁₂O₄₆[M + Na]⁺2067.731; found 2067.826.

Methyl 4,6-di-O-(5-azido-3-oxapentyl)-2,3-di-O-[1-(2,3,4,6-tetra-O-

acetyl-α- -mannopyranosyloxyethyl)-4-methyl-1H-1,2,3-triazole-

1,4-diyl]-α- -glucopyranoside (78): Obtained from 77 (439 mg) fol-

D

lowing the general procedure for the synthesis of the azido reac-

tions. Column chromatography (EtOAc/MeOH, 15:1) gave 78 as a

syrup (403 mg, 91%): [α]²_D⁰= +57 (c = 1 in chloroform). IR (KBr):

ν = 2110, 1749, 1227, 1138, 1090, 1048 cm^–1. ¹H NMR (CDCl₃,

˜

300 MHz): δ = 7.96, 7.89 (2 s, 2 H), 5.30–5.15 (m, 6 H), 5.02–4.78

(m, 7 H), 4.70–4.50 (m, 4 H), 4.23 (dd, J = 12.4, 5.0 Hz, 2 H),

4.20–3.30 (several m, 28 H), 3.86 (t, J = 9.4 Hz, 1 H), 3.55 (dd, J

= 9.6, 3.5 Hz, 1 H), 3.39 (s, 3 H), 2.17, 2.10, 2.04, 1.98 (4 s, 24 H)

ppm. ¹³C NMR (CDCl₃, 75 MHz): δ = 170.5, 169.8, 169.7, 169.6,

145.4, 144.9, 124.4, 124.1, 97.6, 81.4, 79.5, 78.0, 72.0, 70.8, 70.7,

70.5, 70.0, 69.9, 69.8, 69.5, 69.1, 69.0, 68.8, 68.8, 66.3, 65.7, 64.3,

61.1, 54.9, 50.7, 49.5, 20.7, 20.6, 20.6, 20.5 ppm. HRMS (FAB+):

calcd. for C₅₃H_7lO₂₈N₁₂[M + Na]⁺1353.495; found 1353.454.

1,15-Bis[4-(α-

D-glucopyranosyloxymethyl)-1H-1,2,3-triazol-1-yl]-

8,8-bis[1-(α- -mannopyranosyloxyethyl)-4-methyloxy-1H-1,2,3-tri-

D

azole-1,4-diyl]-3,6,10,13-tetraoxapentadecane (75): Obtained from

73 (409 mg) following the general procedure for the de-O-acety-

Methyl 2,3-Di-O-[1-(2,3,4,6-tetra-O-acetyl-α-

ethyl)-4-methyl-1H-1,2,3-triazole-1,4-diyl]-4,6-di-O-[4-(2,3,4,6-

tetra-O-acetyl-β- -glucopyranosyloxymethyl)-1-(3-oxapentyl)-1H-

1,2,3-triazole-1,4-diyl]-α- -glucopyranoside (79): Obtained from 78

(266 mg) and 3 (185 mg) following the general procedure for the

CuAAC reactions. Column chromatography (EtOAc/MeOH, 10:1)

gave 79 as a foamy solid (382 mg, 91%): [α]²_D⁰= +17 (c = 1 in

D-mannopyranosyloxy-

lation (method A). Column chromatography (MeOH) gave 75 as a

D

20

syrup (273 mg, 98%): [α]²_D⁰= +3, [α] = +9 (c = 0.5 in MeOH).

436

D

IR (KBr): ν = 3390, 2919, 1651, 1479, 1057 cm^–^{1 1}H NMR

.

˜

(CD₃OD, 300 MHz, selected signals): δ = 8.05, 7.99 (2 s, 4 H, H-5

triazole), 4.91 (d, J = 3.6 Hz, 2 H), 4.73 (s, 2 H) ppm. ¹³C NMR

(CD₃OD, 75 MHz): δ = 146.2, 145.3, 126.1, 125.8, 101.6, 99.6, 75.1,

74.9, 74.0, 73.5, 72.5, 72.1, 71.9, 71.8, 71.4, 70.7, 70.4, 70.1, 68.4,

66.8, 65.3, 62.8, 62.7, 61.4, 51.5, 51.3, 46.6 ppm. HMRS (MALDI-

TOF): calcd. for C₅₃H₈₈N₁₂O₃₀[M + Na]⁺1395.560; found

1395.579.

chloroform). IR (KBr): ν = 1754, 1372, 1231, 1139, 1089, 1048

˜

cm^–1. ¹H NMR (CDCl₃, 300 MHz, selected signals): δ = 7.91, 7.89,

7.75, 7.67 (4 s, 2 H), 3.39 (s, 3 H), 2.14, 2.13, 2.10, 2.09, 20.4, 2.03,

2.02, 1.99, 1.97, 1.96, 1.95 (10 s, 48 H) ppm. ¹³C NMR (CDCl₃,

75 MHz): δ = 170.4, 170.4, 170.0, 169.8, 169.6, 169.4, 169.4, 145.0,

144.6, 143.6, 124.3, 124.0, 123.7, 99.6, 97.4, 81.2, 79.2, 77.8, 72.6,

71.7, 71.0, 70.6, 70.3, 69.2, 69.1, 69.0, 68.8, 68.7, 68.2, 66.1, 65.6,

62.6, 62.1, 61.7, 54.9, 50.1, 49.4, 20.6, 20.6, 20.5, 20.4 ppm. HRMS

(FAB+): calcd. for C₈₇H₁₂₂O₄₈N₁₂[M + Na]⁺2125.737; found

2125.736.

1,15-Bis[4-(β-

D-glucopyranosyloxymethyl)-1H-1,2,3-triazol-1-yl]-

8,8-bis[1-(α- -mannopyranosyloxyethyl)-4-methyloxy-1H-1,2,3-tri-

D

azole-1,4-diyl]-3,6,10,13-tetraoxapentadecane (76): Obtained from

74 (409 mg) following the general procedure for the de-O-acety-

lation (method A). Column chromatography (MeOH) gave 76 as a

syrup (257 mg, 92%): [α]²_D⁰= +64 (c = 1 in MeOH). IR (KBr): ν =

˜

Methyl 4,6-Di-O-[4-(β-

1H-1,2,3-triazole-1,4-diyl]-2,3-Di-O-[1-(α-

ethyl)-4-methyl-1H-1,2,3-triazole-1,4-diyl]-α-

D

-glucopyranosyloxymethyl)-1-(3-oxapentyl)-

-mannopyranosyloxy-

-glucopyranoside

1

3365, 2908, 1654, 1458, 1046 cm^–1. H NMR (CD₃OD, 300 MHz,

D

selected signals): δ = 8.02, 7.98 (2 s, 4 H), 4.40 (d, J = 7.7 Hz, 2 H)

ppm. ¹³C NMR (CD₃OD, 75 MHz): δ = 146.2, 145.5, 126.1, 125.8,

103.6, 101.6, 78.0, 75.0, 74.9, 72.5, 72.1, 71.9, 71.6, 71.4, 70.4, 70.0,

68.4, 66.8, 65.3, 63.0, 62.8, 51.5, 51.2 ppm. HMRS (MALDI-

TOF): calcd. for C₅₃H₈₈N₁₂O₃₀[M + Na]⁺1395.56; found 1395.55.

D

(80): Obtained from 79 (350 mg) following the general procedure

for the de-O-acetylation (method B). Column chromatography

(MeOH) gave 80 as a syrup (212 mg, 89%): [α]²_D⁰= –9 (c = 0.25 in

H O). IR (KBr): ν = 3400, 2924, 1135, 1055 cm^{–1 1}H NMR

.

˜

2

(CD₃OD, 300 MHz, selected signals): δ = 8.10, 8.06, 8.03, 8.01 (4

s, 4 H) ppm. ¹³C NMR (CD₃OD, 75 MHz): δ = 146.4, 145.9, 145.6,

126.2, 126.1, 126.0, 103.8, 103.6, 101.6, 99.0, 82.5, 80.7, 79.1, 78.0,

75.0, 74.9, 73.0, 72.5, 72.4, 71.8, 71.7, 71.5, 71.4, 70.3, 70.2, 68.4,

67.0, 66.8, 66.7, 64.8, 63.2, 63.1, 62.8, 55.7, 51.5, 51.2, 49.9 ppm.

HRMS (MALDI-TOF): calcd. for C₅₅H₉₀O₃₂N₁₂[M + Na]⁺

1453.60; found 1453.60.

Methyl 4,6-Di-O-(5-chloro-3-oxapentyl)-2,3-di-O-[1-(2,3,4,6-tetra-

O-acetyl-α-

1,4-diyl]-α-

D

-mannopyranosyloxyethyl)-4-methyl-1H-1,2,3-triazole-

D-glucopyranoside (77): Obtained from 62 (241 mg) and

7 (500 mg) following the general procedure for the CuAAC reac-

tions. Column chromatography (EtOAc/MeOH, 15:1) gave 77 as a

syrup (526 mg, 80%): [α]²_D⁰= +50 (c = 1 in chloroform). IR (KBr):

ν = 1752, 1228, 1139, 1091, 1049 cm^{–1 1}H NMR (CDCl₃,

.

˜

300 MHz): δ = 7.99, 7.92 (2 s, 2 H, H-5 triazole), 5.30–5.19 (m, 6

H, H-2,3,4), 4.98 (AB system, J = 11.4 Hz, ∆µ = 13.0 Hz, 2 H,

OCH₂-triazole), 4.84 (d, J = 12.2 Hz, 1 H, OCH₂-triazole), 4.83 (d,

J = 3.8 Hz, 1 H, H-1 Glc), 4.80–4.79 (2 s, 2 H, H-1 Man), 4.81 (d,

Methyl 3,4-Di-O-(5-chloro-3-oxapentanyl)-2,6-di-O-[1-(2,3,4,6-

tetra-O-acetyl-α-

azole-1,4-diyl]-α-

(241 mg) and 7 (500 mg) following the general procedure for the

D-mannopyranosyloxyethyl)-4-methyl-1H-1,2,3-tri-

D

-galactopyranoside (81): Obtained from 66

J = 11.9 Hz, 1 H, OCH₂-triazole), 4.67–4.50 (m, 4 H, CH₂N), 4.27– CuAAC reactions. Column chromatography (EtOAc/MeOH, 15:1)

3.88 (m, 8 H, H-6,6Ј Man, CH₂O-Man), 3.84 (t, J = 9.4 Hz, 1 H,

gave 81 as a syrup (618 mg, 94%): [α]²_D⁰= +42 (c = 1 in chloroform).

H-3 Glc), 3.80–3.57 (m, 21 H, H-5,6,6’ Glc, H-5 Man, ClCH₂CH₂- IR (film): ν = 1751, 1228, 1139, 1092, 1048 cm^–1. ¹H NMR (CDCl ,

˜

3

OCH₂CH₂O), 3.53 (dd, J = 9.6, 3.5 Hz, 1 H, H-2 Glc), 3.42 (t, J 300 MHz): δ = 7.74 (s, 1 H), 7.69 (s, 1 H), 5.27–5.20 (m, 6 H), 4.96

= 9.2 Hz, 1 H, H-4 Glc), 3.38 (s, 3 H, OMe), 2.12, 2.08, 2.03, 1.97

(d, J = 12.1 Hz, 1 H), 4.83 (d, J = 12.1 Hz, 1 H), 4.82 (d, J =

2470

www.eurjoc.org

Eur. J. Org. Chem. 2009, 2454–2473

Click Multivalent Heterogeneous Neoglycoconjugates

3.7 Hz, 1 H), 4.80 (br. s, 2 H), 4.73 (d, J = 12.5 Hz, 1 H), 4.67 (d,

J = 12.5 Hz, 1 H), 4.65–4.55 (m, 4 H), 4.27–3.55 (m, 28 H), 3.37

(s, 3 H), 2.14, 2.09, 2.05, 1.99 (4 s, 24 H) ppm. ¹³C NMR (CDCl₃,

75 MHz): δ = 170.4, 169.7, 169.7, 169.4, 145.5, 145.0, 123.9, 123.7,

98.4, 97.5, 97.4, 79.4, 76.0, 72.3, 71.1, 71.0, 70.8, 70.2, 69.0, 68.9,

68.8, 68.7, 66.2, 66.1, 65.6, 64.6, 64.4, 62.1, 55.1, 49.6, 49.5, 43.1,

43.0, 20.7, 20.6, 20.5 ppm. HMRS (MALDI-TOF): calcd. for

C₅₃H₇₈N₆O₂₈Cl₂[M + Na]⁺1318.115; found 1339.438.

101.6, 99.7, 80.4, 78.0, 78.0, 77.2, 76.9, 75.0, 74.9, 73.2, 72.4, 72.0,

71.9, 71.8, 71.8, 71.6, 71.0, 60.4, 70.2, 70.2, 70.1, 68.4, 68.3, 66.7,

65.1, 65.0, 63.1, 63.0, 62.8, 55.7, 51.5, 51.2, 49.6 ppm. MS

(MALDI-TOF): calcd. for C₅₅H₉₀N₁₂O₃₂[M + Na]⁺1453.568;

found 1453.67.

4,6:4Ј,6Ј-Tetra-O-(5-chloro-3-oxapentyl)-2,3,2Ј,3Ј-tetra-O-[1-

(2,3,4,6-tetra-O-acetyl-α-D-mannopyranosyloxyethyl)-4-methyl-1H-

1,2,3-triazole-1,4-diyl]-α,αЈ-trehalose (85): Obtained from 70

(230 mg) and 7 (500 mg) following the general procedure for the

CuAAC reactions. Column chromatography (EtOAc/MeOH, 15:1)

Methyl 3,4-Di-O-(5-azido-3-oxapentyl)-2,6-di-O-[1-(2,3,4,6-tetra-O-

acetyl-α-

1,4-diyl]-α-

D

-mannopyranosyloxyethyl)-4-methyl-1H-1,2,3-triazole-

-galactopyranoside (82): Obtained from 81 (439 mg)

D

gave 85 as a syrup (595 mg, 92%): [α]²_D⁰= +65 (c = 1 in chloroform).

1

following the general procedure for the synthesis of the azido deriv-

atives. Column chromatography (EtOAc/MeOH, 15:1) gave 82 as a

foamy solid (333 mg, 75%): [α]²_D⁰= +41.5 (c = 1 in chloroform). IR

IR (film): ν = 1751, 1372, 1228, 1139, 1091, 1049 cm^–1. H NMR

˜

(CDCl₃, 300 MHz, selected signals): δ = 8.09 (s, 2 H), 7.89 (s, 2 H),

2.14, 2.11, 2.10, 2.05, 2.04, 1.98 (6 s, 48 H) ppm. ¹³C NMR (CDCl₃,

(KBr): ν = 2211, 1749, 1228, 1139, 1092, 1048 cm^–1. ¹H NMR 75 MHz): δ = 170.6, 169.8, 169.7, 169.6, 145.3, 144.9, 124.5, 124.3,

˜

(CDCl₃, 300 MHz): δ = 7.71 (s, 1 H), 7.68 (s, 1 H), 5.28–5.19 (m, 97.6, 93.4, 80.6, 78.8, 77.8, 71.9, 71.1, 71.0, 70.7, 70.6, 70.3, 69.4,

6 H), 4.91 (d, J = 12.2 Hz, 1 H), 4.83 (d, J = 12.2 Hz, 1 H), 4.82

(d, J = 3.6 Hz, 1 H), 4.79 (br. s, 2 H), 4.73 (d, J = 12.2 Hz, 1 H),

4.67 (d, J = 12.2 Hz, 1 H), 4.64–4.52 (m, 4 H), 4.25–3.59 (m, 24

H), 3.41–3.35 (m, 4 H), 3.37 (s, 3 H), 2.14, 2.09, 2.05, 1.99 (4 s, J

= 24 Hz) ppm. ¹³C NMR (CDCl₃, 75 MHz): δ = 170.5, 169.9,

169.8, 169.6, 159.6, 145.6, 145.2, 123.9, 123.9, 98.6, 97.6, 97.5, 79.6,

76.2, 76.2, 72.5, 71.0, 70.9, 70.3, 69.0, 69.2, 69.1, 69.0, 69.0, 68.9,

68.8, 66.3, 66.2, 65.8, 65.8, 64.8, 64.6, 62.2, 55.3, 50.9, 50.8, 49.7,

49.6, 20.8, 20.7, 20.6 ppm. HMRS (MALDI-TOF): calcd. for

C₅₃H₇₈N₁₂O₂₈[M + Na]⁺1331.495; found 1353.517.

69.0, 68.9, 68.8, 68.7, 68.7, 66.3, 66.2, 66.0, 65.6, 63.8, 62.2, 49.4,

49.3, 43.2, 42.8, 20.7, 20.6, 20.5 ppm. HRMS (MALDI-TOF):

calcd. for C₁₀₄H₁₅₀Cl₄O₅₅N₁₂[M + Na]⁺2609.812; found 2609.880.

4,6:4Ј,6Ј-Tetra-O-(5-azido-3-oxapentyl)-2,3,2Ј,3Ј-tetra-O-[1-

(2,3,4,6-tetra-O-acetyl-α-D-mannopyranosyloxyethyl)-4-methyl-1H-

1,2,3-triazole-1,4-diyl]-α,αЈ-trehalose (86): Obtained from 85

(517 mg) following the general procedure for the synthesis of the

azido reactions. Column chromatography (EtOAc/MeOH, 10:1)

gave as a solid foamy 86 (418 mg, 80%): [α]²_D⁰= +68 (c = 1 in

chloroform). IR (KBr): ν = 2113, 1751, 1227, 1139, 1090, 1048

˜

1

Methyl 3,4-Di-O-[1-(2,3,4,6-tetra-O-acetyl-β-

methyl)-1-(3-oxapentyl)-1H-1,2,3-triazole-1,4-diyl]-2,6-di-O-[1-

(2,3,4,6-tetra-O-acetyl-α- -mannopyranosyloxyethyl)-1H-1,2,3-tri-

azole-1,4-diyl]-α- -galactopyranoside (83): Obtained from 82

D-glucopyranosyloxy-

cm^–1. H NMR (CDCl₃, 300 MHz): δ = 8.09, 7.89 (2 s, 4 H, H-5

triazole), 5.30–5.18 (m, 14 H, H-1 trehalose, H-2,3,4 Man), 5.00,

4.95 (2 d, J = 11.3 Hz, 4 H, CH₂-triazole), 4.85, 4.82 (2 br. s, 4 H,

H-1 Man), 4.83, 4.68 (2 d, 4 H, CH₂- triazole), 4.70–4.50 (m, 8 H,

CH₂N triazole), 4.25 (m, 4 H, H-6 Man), 4.15 (m, 4 H, CH₂O-

Man), 4.08 (bd, 4 H, H-6’ Man), 4.08–3.90 (m, 6 H, CH₂Man, H-

5 trehalose), 3.85 (t, J = 9.3 Hz, 2 H, H-3 trehalose), 3.85–3.60 (m,

32 H, CH₂O spacer, H-6 trehalose, H-5 Man), 3.51 (dd, J = 9.6,

3.5 Hz, 2 H, H-2 trehalose), 3.47–3.30 (m, 10 H, CH₂N₃, H-4 treha-

lose), 2.13–1.97 (5 s, 48 H, CH₃CO) ppm. ¹³C NMR (CDCl₃,

D

(266 mg) and 3 (185 mg) following the general procedure for the

CuAAC reactions. Column chromatography (EtOAc/MeOH, 10:1)

20

gave 83 as a foamy solid (395 mg, 94%): [α]²_D⁰= +9, [α] = +16 (c

436

= 1 in chloroform). IR (KBr): ν = 1755, 1231, 1138, 1092, 1049

˜

1

cm^–1. H NMR (CDCl₃, 400 MHz): δ = 7.75, 7.68, 7.66, 7.63 (4 s,

4 H, H-5 triazole), 5.23 (t, J = 9.7 Hz, 2 H, H-4 Man), 5.20–5.14

(m, 6 H, H-2,3 Man, H-3 Glc), 5.07 (br. t, J = 9.6 Hz, 2 H, H-4 75 MHz): δ = 170.6, 169.9, 169.8, 169.7 (CH₃CO), 145.6, 145.2 (C-

Glc), 4.96 (dd, J = 9.2, 8.0 Hz, 2 H, H-2 Glc), 4.88 (d, J = 12.5 Hz,

1 H, OCH₂-triazole), 4.87 (d, J = 12.5 Hz, 1 H, OCH₂-triazole),

4.86 (d, J = 12.1 Hz, 1 H, OCH₂-triazole), 4.78 (d, J = 3.5 Hz, 1

H, H-1 Gal), 4.77 (br. s, 2 H, H-1 Man), 4.76 (d, J = 12.1 Hz, 1

H, OCH₂-triazole), 4.75 (d, J = 12.5 Hz, 1 H, OCH₂-triazole), 4.74

(d, J = 12.5 Hz, 1 H, OCH₂-triazole), 4.69 (d, J = 8.0 Hz, 1 H, H-

1 Glc), 4.68 (d, J = 8.0 Hz, 1 H, H-1 Glc), 4.65 (d, J = 12.4 Hz, 1

H, OCH₂-triazole), 4.58 (d, J = 12.4 Hz, 1 H, OCH₂-triazole),

4 triazole), 124.5, 124.2 (C-5 triazole), 97.8 (C-1 Man), 93.6 (C-

1 trehalose), 81.0 (C-3 trehalose), 79.0 (C-2 trehalose), 78.1 (C-4

trehalose), 72.1, 71.0, 70.9, 70.8 (C-5 trehalose), 70.5, 69.9, 69.8

(CH₂spacer), 69.5 (C-6 trehalose), 69.2, 69.0, 68.9 (C-2,3,5 Man),

66.5, 66.4 (CH₂Man), 66.3 (CH₂triazole), 65.9 (C-4 Man), 64.1

(CH₂triazole), 62.3 (C-6 Man), 50.8, 50.7 [CH₂(N) triazole], 49.5,

49.4 (CH₂N₃), 20.9, 20.8, 20.8, 20.7 (CH₃COO) ppm. HRMS

(MALDI-TOF): calcd. for C₁₀₄H₁₅₀O₅₅N₂₄[M + Na]⁺2637.973;

4.60–4.45 (m, 8 H, 4 CH₂N), 4.27 (dd, J = 12.4, 4.5 Hz, 1 H, H-6 found 2637.790.

Glc), 4.26 (dd, J = 12.4, 4.5 Hz, 1 H, H-6 Glc), 4.23–4.00, 3.95–

2,3,2Ј,3Ј-Tetra-O-[1-(2,3,4,6-tetra-O-acetyl-α-

ethyl)-4-methyl-1H-1,2,3-triazole-1,4-diyl]-4,6:4Ј,6Ј-tetra-O-[4-

(2,3,4,6-tetra-O-acetyl-α- -glucopyranosyloxymethyl)-1-(3-oxapen-

D-mannopyranosyloxy-

3.50 [2 m, 40 H, H-2,3,4, H-5,6,6’ Gal, (CH₂CH₂)₂O, triazole-

CH₂CH₂O, H-5 Glc, H-5 Man, H-6 Glc, 4 H-6 Man], 3.37 (s, 3 H,

OMe), 2.13, 2.09, 2.04, 2.02, 1.99, 1.98, 1.96 (7 s, 48 H, 16 Ac)

ppm. HRMS (MALDI-TOF): calcd. for C₈₇H₁₂₂N₁₂O₄₈[M + Na]

D

tyl)-1H-1,2,3-triazole-1,4-diyl]-α,αЈ-trehalose (87): Obtained from 2

(309 mg) and 86 (436 mg) following the general procedure for the

CuAAC reactions. Column chromatography (EtOAc/MeOH, 10:1)

gave 87 as a foamy solid (527 mg, 76%): [α]²_D⁰= +88 (c = 1 in

+

2125.737; found 2125.696.

Methyl 3,4-Di-O-[1-(β-

1H-1,2,3-triazole-1,4-diyl]-2,6-di-O-[1-(α-

ethyl)-1H-1,2,3-triazole-1,4-diyl]-α- -galactopyranoside (84): Ob-

D-glucopyranosyloxymethyl)-1-(3-oxapentyl)-

D

-mannopyranosyloxy-

chloroform). IR (film): ν = 1751, 1371, 1229, 1046 cm^–1. ¹H NMR

˜

D

(CDCl₃, 300 MHz, selected signals): δ = 8.10, 7.88, 7.79, 7.63 (4 s,

4 H, H-5 triazole), 5.46, 5.44 (2 t, J = 9.8 Hz, 2 H, H-3 Glc), 5.3–

5.15 (m), 5.08 (t, J = 9.8 Hz, 2 H, H-4 Glc), 5.00–4.57 (m), 4.85 (s,

2 H, H-1 Man), 4.52 (t, J = 5.0 Hz, 2 H, CH₂), 4.30–3.35 (several

tained from 83 (350 mg) following the general procedure for the

de-O-acetylation (method B). Column chromatography (MeOH)

gave 84 as a syrup (219 mg, 92%): [α]²_D⁰= +10 (c = 0.25 in H₂O).

IR (KBr): ν = 3395, 2924, 1135, 1090, 1055 cm^{–1 1}H NMR m), 2.14, 2.13, 2.09, 2.04, 2.04, 2.02, 1.99, 1.98, 1.96 (several s, 96

.

˜

(CD₃OD, 300 MHz, selected signals): δ = 8.08, 8.04, 8.01, 7.97 (4

s, 4 H), 3.34 (s, 3 H) ppm. ¹³C NMR (CD₃OD, 100 MHz): δ = 170.0, 169.9, 169.5 (COO), 145.1, 144.7, 143.3 (C-4 triazole), 124.6,

146.1, 145.8, 145.5, 145.5, 126.2, 126.1, 126.0, 126.0, 103.7, 103.6, 124.1, 123.9 (C-5 triazole), 97.6, 97.5 (C-1 Man), 95.1 (C-1Glc),

H, 32 Ac) ppm. ¹³C NMR (CDCl₃, 75 MHz): δ = 170.6, 170.5,

Eur. J. Org. Chem. 2009, 2454–2473

www.eurjoc.org

2471

F. Santoyo-Gonzalez et al.

FULL PAPER

93.2 (C-1 trehalose), 80.7, 78.8, 77.9, 71.8, 71.1, 70.7, 70.5, 70.3,

70.0, 69.3, 69.1, 68.9, 68.8, 68.4, 67.4, 66.2, 65.7, 63.8, 62.1, 61.6,

61.2, 61.1, 60.3, 50.1, 49.5, 49.4 (CH₂N), 20.7, 20.6, 20.6 (CH₃CO)

ppm. HRMS (MALDI-TOF): calcd. for C₁₇₂H₂₃₈N₂₄O₉₅[M +

Na]⁺4184.885; found 4184.896.

clusters or methyl α--Man and the peroxidase-labeled lectin

(100 µL/well) were added, and the plates were incubated for 2 h at

37 °C. After that, 50 µL/well of a solution of o-phenylenediamine

dihydrochloride (20 mg/50 mL) in citrate–phosphate buffer (pH 5.0

with 0.4% H₂O₂) was added. The plates were incubated for 30 min

at 37 °C. The reactions were stopped by addition of aqueous

H₂SO₄(50 µL/well, 1.25 ), and the absorbance measured at

492 nm.

2,3,2Ј,3Ј-Tetra-O-[1-(2,3,4,6-tetra-O-acetyl-α-

ethyl)-4-methyl-1H-1,2,3-triazole-1,4-diyl]-4,6:4Ј,6Ј-tetra-O-[4-

(2,3,4,6-tetra-O-acetyl-β- -glucopyranosyloxymethyl)-1-(3-oxapen-

D-mannopyranosyloxy-

D

tyl)-1H-1,2,3-triazole-1,4-diyl]-α,αЈ-trehalose (88): Obtained from 3

(309 mg) and 86 (436 mg) following the general procedure for the

CuAAC reactions. Column chromatography (EtOAc/MeOH, 10:1)

gave 79 as a foamy solid (596 mg, 86%): [α]²_D⁰= +20 (c = 1 in

Acknowledgments

chloroform). IR (film): ν = 1751, 1371, 1228, 1139, 1089, 1046

˜

We thank Dirección General de Investigación Científica y Técnica

cm^–1. ¹H NMR (CDCl₃, 300 MHz, selected signals): δ = 8.09, 7.88,

7.75, 7.70 (4 s, 8 H), 2.14, 2.13, 2.10, 2.09, 2.08, 2.05, 2.04, 2.03,

1.99, 1.97, 1.96, 1.95 (several s, 96 H) ppm. ¹³C NMR (CDCl₃,

75 MHz): δ = 170.6, 170.1, 169.9, 169.6, 169.4, 169.3, 145.1, 144.7,

143.7, 143.7, 124.6, 124.2, 123.9, 99.8, 99.7, 97.5, 93.0, 80.5, 72.7,

71.8, 71.1, 70.7, 70.3, 69.2, 69.0, 68.9, 68.8, 68.3, 66.2, 65.7, 62.7,

62.1, 61.8, 50.2, 49.4, 20.7, 20.6, 20.5 ppm. HRMS (MALDI-

TOF): calcd. for C₁₇₂H₂₃₈N₂₄O₉₅[M + Na]⁺4184.885; found

4184.896.

(DGICYT) for financial support (CTQ2005-02219).

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2.

2,3,2Ј,3Ј-Tetra-O-[1-(α-

1,2,3-triazole-1,4-diyl]-4,6:4Ј,6Ј-tetra-O-[4-(α-

D

-mannopyranosyloxyethyl)-4-methyl-1H-

-glucopyranosyloxy-

D

methyl)-1-(3-oxapentyl)-1H-1,2,3-triazole-1,4-diyl]-α,αЈ-trehalose

(89): Obtained from 87 (520 mg) following the general procedure

for the de-O-acetylation (method B). Column chromatography

(acetonitrile/H₂O, 2:1) gave 89 as a syrup (303 mg, 86%): [α]²_D⁰

=

1

+76 (c = 0.25 in H O). IR (film): ν = 3406, 1104 cm^–1. H NMR

˜

2

([D₆]DMSO, 300 MHz, selected signals): δ = 8.09, 8.06, 8.04, 8.01

(4 s, 8 H) ppm. ¹³C NMR ([D₆]DMSO, 100 MHz): δ = 145.2, 144.8,

144.4, 125.4, 125.3, 1235.2, 100.3, 98.6, 93.0, 81.2, 79.0, 77.9, 77.2,

74.3, 73.9, 73.2, 72.4, 71.4, 70.7, 70.1, 69.2, 67.3, 66.1, 65.6, 61.7,

61.5, 60.5, 60.4, 50.4, 50.2 ppm. HRMS (MALDI-TOF): calcd. for

C

₁₀₈H₁₇₄N₂₄O₆₃[M + Na]⁺2838.104; found 2837.929.

2,3,2Ј,3Ј-Tetra-O-[1-(α- -mannopyranosyloxyethyl)-4-methyl-1H-

1,2,3-triazole-1,4-diyl]-4,6:4Ј,6Ј-tetra-O-[4-(β- -glucopyranosyloxy-

D

methyl)-1-(3-oxapentyl)-1H-1,2,3-triazole-1,4-diyl]-α,αЈ-trehalose

(90): Obtained from 88 (520 mg) following the general procedure

for the de-O-acetylation (method B). Column chromatography

(acetonitrile/H₂O, 2:1) gave 90 as a syrup (281 mg, 80%): [α]²_D⁰

=

+8.7, [α]²⁰= +15.3 (c = 0.25 in H O). IR (film): ν = 3384, 1103

˜

436

2

1

cm^–1. H NMR ([D₆]DMSO, 300 MHz, selected signals): δ = 8.10,

8.07, 8.06, 8.01 (4 s, 8 H) ppm. ¹³C NMR ([D₆]DMSO, 100 MHz):

δ = 144.3, 143.9, 143.6, 124.5, 124.2, 124.0, 102.1, 99.8, 76.9, 76.7,

74.1, 74.0, 73.3, 70.8, 70.1, 70.0, 69.2, 68.6, 68.5, 66.8, 64.8, 63.2,

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C₁₀₈H₁₇₄N₂₄O₆₃[M + Na]⁺2838.104; found 2838.002.

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Enzyme-Linked Lectin Assay (ELLA): ELLA assays were carried

out as described previously.^[33]Experiments were carried out with

a Metertech Σ960 instrument. Microtitration plates were coated

with S. cerevisiae mannan at 100 µL/well of a solution of 10 µg/mL

in 10 m phosphate buffer (PBS, pH 7.4) for 2 h at 37 °C. The

wells were then washed twice with 10 m phosphate buffer contain-

ing 1% (v/v) Tween 20 (PBST) and once with PBS. This washing

procedure was repeated after each incubation period. Wells were

then blocked with 300 µL/well of BSA/PBS (1% w/v) for 2 h at

37 °C. Each inhibitor was added in serial dilutions (60 µL/well) of

the glycoconjugates or methyl α--Man in PBS (pH 6.8, containing

0.1 m Ca²⁺and 0.1 m Mn²⁺), and the peroxidase-labeled Con A

(60 µL/well of a solution of 50 µg/mL in PBS, pH 6.8, containing

0.1 m Ca²⁺and 0.1 m Mn²⁺) was added. The mixtures of glyco-

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www.eurjoc.org

Eur. J. Org. Chem. 2009, 2454–2473

Click Multivalent Heterogeneous Neoglycoconjugates

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Received: November 25, 2008

Published Online: April 2, 2009

Eur. J. Org. Chem. 2009, 2454–2473

www.eurjoc.org

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Click multivalent heterogeneous neoglycoconjugates - modular synthesis and evaluation of their binding affinities

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