Design, synthesis and biological evaluation of mannosyl triazoles as FimH antagonists

Bioorganic & Medicinal Chemistry 19 (2011) 6454–6473

Contents lists available at SciVerse ScienceDirect

Bioorganic & Medicinal Chemistry

journal homepage: w ww.elsevier.com/locate/bmc

Design, synthesis and biological evaluation of mannosyl triazoles

as FimH antagonists

Oliver Schwardt, Said Rabbani, Margrit Hartmann, Daniela Abgottspon, Matthias Wittwer,

⇑

Simon Kleeb, Adam Zalewski, Martin Smieško, Brian Cutting, Beat Ernst

Institute of Molecular Pharmacy, Pharmacenter, University of Basel, Klingelbergstrasse 50, CH-4056 Basel, Switzerland

a r t i c l e i n f o

a b s t r a c t

Article history:

Urinary tract infection (UTI) caused by uropathogenic Escherichia coli (UPEC) is one of the most prevalent

infectious diseases. Particularly affected are women, who have a 40–50% risk to experience at least one

symptomatic UTI episode at some time during their life. In the initial step of the infection, the lectin

FimH, located at the tip of bacterial pili, interacts with the high-mannosylated uroplakin Ia glycoprotein

on the urinary bladder mucosa. This interaction is critical for the ability of UPEC to colonize and invade

the bladder epithelium. X-ray structures of FimH co-crystallized with two different ligands, the physio-

Received 22 April 2011

Revised 24 August 2011

Accepted 25 August 2011

Available online 31 August 2011

Keywords:

logical binding epitope oligomannose-3 and the antagonist biphenyl a-D-mannoside 4a revealed different

Uropathogenic Escherichia coli

Urinary tract infections

Bacterial adhesin FimH

FimH antagonists

Competitive binding assay

Aggregometry assay

NMR spectroscopy

binding modes, an in-docking-mode and an out-docking-mode, respectively. To accomplish the in-docking-

mode, that is the docking mode where the ligand is hosted by the so-called tyrosine gate, FimH antago-

nists with increased ﬂexibility were designed and synthesized. All derivatives 5–8 showed nanomolar

afﬁnities, but only one representative, the 4-pyridiyl derivative 5j, was as potent as the reference com-

pound n-heptyl

a-D-mannoside (1b). Furthermore, a loss of afﬁnity was observed for C-glycosides and

derivatives where the triazole aglycone is directly N-linked to the anomeric center. A conformational

analysis by NMR revealed that the triazolyl-methyl-C-mannosides 8 adopt an unusual ¹C₄chair confor-

mation, explaining the comparably lower afﬁnity of these compounds. Furthermore, to address the drug-

likeness of this new class of FimH antagonists, selected pharmacokinetic parameters, which are critical

for oral bioavailability (lipophilicity, solubility, and membrane permeation), were determined.

1. Introduction

ported cases. Particularly affected are women, who have a 40–

50% risk to suffer from a symptomatic UTI episode at some time

Urinary tract infections (UTIs) are among the most common

infections, affecting millions of people each year. Although UTIs

rarely cause severe diseases such as pyelonephritis or urosepsis,

they are associated with extensive morbidity and generate consid-

erable medical expenses.¹Uropathogenic Escherichia coli (UPEC)

are the primary cause of UTIs accounting for 70–95% of the re-

during their life.^2,3Symptomatic UTIs require antimicrobial treat-

ment, resulting in selection and development of bacterial resis-

tance. Consequently, treatment of consecutive infections becomes

increasingly difﬁcult. Especially patients with diabetes, urinary

tract anomaly, paraplegia and those with permanent urinary cath-

eter experience repeated UTIs with resistant strains. Therefore, a

new approach for the treatment and prevention of UTI with non-

antibiotic and orally applicable therapeutics with a low potential

for resistance would have a great impact on patient care, public

healthcare, and medical expenses.

UPEC express a number of well-studied virulence factors for

successful colonization of and survival within the host.^1,4,5One

important virulence factor, the mannose-speciﬁc FimH adhesin, is

located at the tip of bacterial type 1 pili.⁶Type 1 pili are the most

prevalent ﬁmbriae encoded by UPEC, consisting of the four sub-

units FimA, FimF, FimG and FimH. The FimH lectin enables UPEC

to attach to high-mannosylated uroplakin Ia glycoproteins on the

urinary bladder mucosa, thus enabling adherence and invasion of

host cells and at the same time preventing the rapid clearance of

Abbreviations: ABTS, 2,2⁰-azino-di-(3-ethylbenzthiazoline-6-sulfonic acid); AUC,

area under the curve; BSA, bovine serum albumin; CRD, carbohydrate recognition

domain; D, distribution coefﬁcient; DCM, dichloromethane; DMSO, dimethyl

sulfoxide; GIT, gastrointestinal tract; GPE, guinea pig erythrocytes; HEPES, 4-(2-

hydroxyethyl)-piperazine-1-ethanesulfonic acid; IC₅₀

concentration; iv, intravenous; Man, -mannose; NMR, nuclear magnetic reso-

nance; NOESY, nuclear Overhauser enhancement spectroscopy; PAA, polyacryl-

amide; PAMPA, parallel artiﬁcial membrane permeation assay; P_appapparent

permeability; P_e, effective permeation; po, peroral; rIC₅₀relative inhibitory

concentration; SAR, structure–activity relationship; THF, tetrahydrofurane; TM-

PAA, Man (1-3)-[Man (1-6)]-Manb(1-4)-GlcNAcb(1-4)-GlcNAcb-PAA; UPEC, uro-

, half maximal inhibitory

D

,

a

pathogenic E. coli; UTI, urinary tract infection.

⇑

Corresponding author. Tel.: +41 61 2 67 15 51; fax: +41 61 2 67 15 52.

E-mail address: beat.ernst@unibas.ch (B. Ernst).

doi:10.1016/j.bmc.2011.08.057

O. Schwardt et al. / Bioorg. Med. Chem. 19 (2011) 6454–6473

6455

E. coli from the UTI by the bulk ﬂow of urine.^1,7As a part of the

FimH subunit, carbohydrate-recognition domain (CRD) is

responsible for bacterial interactions with the host cells within

the urinary tract.⁷The crystal structure of methyl

-mannopy-

ranoside bound to the FimH-CRD was solved⁸and the structures

of the corresponding complexes with n-butyl -mannopyrano-

(1-6)]-Manb(1-4)-GlcNAcb-(1-4)GlcNAc

hamper an optimal ﬁt with the tyrosine gate.¹¹To increase the con-

formational ﬂexibility, the spacers between the mannose moiety

and the ﬁrst aromatic ring of the biphenyl moiety in i (Fig. 2) as

well as between the aromatic rings was extended. Furthermore,

the rotational barrier of the biphenyl²⁵was reduced by replacing

one of the rings by a triazole (for the torsion proﬁle see Fig. 2).

Overall, these modiﬁcations should lead to a reduction of the con-

formational restraints and therefore an optimized spatial arrange-

ment of the aglycone in the tyrosine gate.

a

-D

a-D

side,⁹Man

a(1-3)-[Mana

(oligomannose-3)¹⁰and biphenyl

became available.

a-D

-mannopyranoside¹¹recently

Previous studies showed that colonization and subsequent

E. coli infection of the human urothelium can be prevented by vac-

cination with FimH adhesin.^12,13Furthermore, adherence and inva-

sion of host cells by E. coli can also be inhibited by oligomannosides

Oligomannose-3 is present on the high-mannosylated uroplakin

Ia located on urothelial cells and is supposed to interact with UPEC.

The crystal structure of the FimH-CRD¹⁰complexed with oligo-

mannose-3 (PDB code 2VCO, Fig. 3A) clearly shows the important

role of the tyrosine gate hosting this physiological ligand in the

so-called in-docking-mode. Interestingly, for 4a complexed with

FimH-CRD a different binding mode outside of the tyrosine gate

was reported (out-docking-mode, see Fig. 3B).¹¹In analogy to oligo-

mannose-3, docking of triazole derivative 5b to the crystal struc-

ture of the FimH lectin domain (PDB code 3MCY)¹¹led - as a

result of the increased ﬂexibility of the aglycone - to the in-dock-

ing-mode. Thus, in contrast to the biphenyl aglycone present in

4a, the phenyl-triazole 5b is expected to be hosted by the tyrosine

gate. The three-dimensional structure 5b was generated using

Glide 5.5²⁶and the kinetic stability of the protein–ligand complex

was then assessed with a 2 ns molecular-dynamics simulation

using Desmond.²⁷

representing the glycosylation of uroplakin 1a.¹⁴For some

a-D-

mannosides it was shown that they prevent type 1 pili mediated

adhesion, that is, they do not act by killing or arresting the growth

of the pathogen as antibiotics do. Therefore, the spread of strains

resistant to such agents are expected to be signiﬁcantly delayed

as compared to that of strains resistant to antibiotics.¹⁵In addition,

environmental contamination is less problematic compared to

antibiotics.^15a

More than two decades ago, various oligomannosides and aro-

matic

a-D-mannosides that antagonize type 1 ﬁmbriae-mediated

bacterial adhesion were identiﬁed.^15,16However, for these manno-

sides only weak interactions in the milli- to micromolar range were

observed. To improve their afﬁnity, the multivalent presentation of

the

a

-mannoside epitope,¹⁷and the rational design of ligands

A comparison of the docking modes of oligomannose-3, 4a and

5b reveals that the interaction of the mannose moiety is highly

conserved for all three compounds. However, in contrast to oligo-

mannose-3 and 5b, the biphenyl moiety in 4a is not able to reach

guided by structural information were explored.^9–11Recently, var-

ious reports on high afﬁnity monovalent FimH antagonists were

published.^11,18,19

The CRD of the FimH protein consists of amino acids with

hydrophilic side chains and can therefore establish a perfect

network of hydrogen bonds with the hydroxyl groups at the 2-,

the tyrosine gate due to its rigid structure. Instead, a p-p-stacking

interaction of the second aromatic ring of the biphenyl aglycone

with Tyr48 outside of the tyrosine gate¹¹(out-docking-mode,

Fig. 3B) is achieved by induced ﬁt, that is, a substantial move of

Tyr48. In addition, a further stabilization of the protein–ligand

complex by a hydrogen bond between the ester in the meta-posi-

tion of 4a and the side-chain of Arg98 was assumed.¹¹

Based on these evidences, a library of derivatives according to

the criteria summarized in Figure 2 was designed. Here, we de-

scribe synthesis, biological evaluation, and determination of phar-

macokinetic parameters of triazole derivatives.

3-, 4- and 6-positions of D-mannose. The entrance to this man-

nose-binding pocket, the so-called ‘tyrosine gate’, is shaped by

two tyrosines (Tyr48 and Tyr137), and one isoleucine (Ile52) which

support hydrophobic contacts.²⁰Generally, long chain alkyl and

aryl mannosides (for selected examples see Fig. 1) displayed the

highest afﬁnities.^{8,9,11,16–21}

Recently, we reported the synthesis, the critical pharmacoki-

netic properties and afﬁnity data of low molecular weight

a-D-

mannosides with the ability to block the FimH-mediated bacterial

adhesion in a mouse infection model.¹⁹The orally available, nano-

molar FimH antagonist 4b (Fig. 1) exhibited the potential to reduce

the colony forming units (CFU) in the urine and in the bladder by

two and four orders of magnitude, respectively, demonstrating

the therapeutic potential of this new class of anti-infectives for

the effective treatment of urinary tract infections.

However, a potential drawback of FimH antagonists with agly-

cons consisting of biphenyls directly linked to the carbohydrate

moiety is their limited conformational ﬂexibility, which could

2. Results and discussion

2.1. Synthesis of triazolyl-methyl and -ethyl

mannopyranosides

a-D-

In a ﬁrst approach, the phenyl ring adjacent to the anomeric

center (see Fig. 2) was replaced by a triazolyl-methyl moiety to in-

crease the conformational ﬂexibility. To avoid solubility problems

as well as to take advantage of additional polar interactions, for

OH

O

HO

R

Cl

R¹

EtO

OR

O

R²

R³

N

H

NO₂

O

2a (R: H)^18a,b

2b (R: Cl)^18c

4a (R¹: H, R²: CO₂Me, R³: H)¹¹

4b (R¹: Cl, R²: H, R³: CO₂Me)¹⁹

4c (R¹: Cl, R²: H, R³: CO₂Na)¹⁹

3^18c

1a (R: Me)

1b (R: n-heptyl)⁹

Figure 1. Known alkyl (1) and aryl (2–4) a-D-mannosides exhibiting micro- to nanomolar afﬁnities.

6456

O. Schwardt et al. / Bioorg. Med. Chem. 19 (2011) 6454–6473

Figure 2. Design of FimH antagonists with aglycons of increased ﬂexibility. Spacer elongations and replacement of one phenyl ring by a triazole should reduce the

conformational restraints and lead to an improved ﬁt in the tyrosine gate. The torsion proﬁles for biphenyl and 1-phenyl-1,2,3-triazole were calculated at the B3LYP level of

theory^22,23with 6-31G(d,p) basis set in the gas phase using Gaussian 03.²⁴

Figure 3. (A) & (B) Crystal structures of oligomannose-3 (A, PDB code 2VCO)¹⁰and biphenyl 4a (B, PDB code 3MCY)¹¹bound to the FimH-CRD. C) Automated docking of

triazole 5b into the lectin domain of FimH (PDB code 3MCY).¹¹The images have been generated using VMD.²⁸The ligands are depicted colored by atom (C: dark grey, H:

white, O: red, N: blue); the tyrosine gate (residues Tyr48, Tyr137 and Ile52) is shown in yellow, residue Thr51 in green and residue Arg98 in red. While 4a binds in the out-

docking-mode, compound 5b, like oligomannose-3, is inserted into the tyrosine gate (in-docking-mode).

example, H-bonds with the hydroxyl-groups of Thr51 or Tyr137

(Fig. 3C), the second aromatic ring was substituted with a carbox-

ylate in para- or meta-position (?5a–c, Scheme 1).

the known aryl azides 11a,³⁰11b,³¹and 11c³²in a copper(I)-

catalyzed Huisgen 1,3-dipolar cycloaddition^33,34using tert-buta-

nol/water/THF (1:1:1) as solvent.³⁵The saponiﬁcation of the

anti-substituted triazoles 12a–c yielded the test compounds 5a–c

(Table 1).

For the synthesis of mannosyl triazoles 5a–c, alkyne 10²⁹read-

ily available from peracetylated D-mannose (9) was reacted with

O. Schwardt et al. / Bioorg. Med. Chem. 19 (2011) 6454–6473

6457

X

N₃

R¹

OAc

R²

Ref.²⁹

11a-c

a)

O

R²

AcO

N

OAc

N

O

R¹

X

12a (X: CH, R¹: CO₂Me, R²: H)

12b (X: CH, R¹: H, R²: CO₂Et)

12c (X: N, R¹: H, R²: CO₂Me)

9

10

OH

O

b)

R²

HO

N

O

R¹

X

5a (X: CH, R¹: CO₂Na, R²: H)

5b (X: CH, R¹: H, R²: CO₂Na)

5c (X: N, R¹: H, R²: CO₂Na)

Scheme 1. Reagents and conditions: (a) CuSO₄ꢀ5H₂O, Na-ascorbate, t-BuOH/H₂O/THF (1:1:1), rt, 24 h, 73–97%; (b) (i) NaOMe, MeOH, rt, 3 h; (ii) 1 M NaOH, H₂O/dioxane (1:1),

16 h, 78–91%.

OAc

OH

Ref.³⁷

R-N₃(15d-i)

O

AcO

HO

N

a)

N

R

O

n

10²⁹(n = 1)

13³⁶(n = 2)

14

5d-i

d: R = CH₂Ph

e: R = 4-NH₂Ph

f: R = adamantyl

R-N₃

b)

(15h-m)

OH

g: R = CH₂(4-MeOPh)

h: R = CH₂(3-MeOPh)

i: R = 4-NO₂Ph

j: R = 4-pyr

OAc

c)

O

HO

O

AcO

N

R

N

O

R

O

n

k: R = 4-F-Ph

n

l: R = 3-F-Ph

m: R = 4-MeOPh

16j-m (n = 1)

17h-k (n = 2)

5j-m (n = 1)

6h-k (n = 2)

Scheme 2. Reagents and conditions: (a) CuSO₄ꢀ5H₂O, Na-ascorbate, t-BuOH/H₂O (1:1), rt, 24 h, (5d–i: 27–77%); (b) CuSO₄ꢀ5H₂O, Na-ascorbate, t-BuOH/H₂O/THF (1:1:1), rt,

24 h (16j–m: 85–94%, 17h–k: 83–96%); (c) NaOMe, MeOH, rt, 2–6 h, (5j–m: 75–85%, 6h–k: 73–90%).

In a second approach, the terminal aromatic ring was replaced

by various substituents like (hetero)aryl, benzyl, and adamantyl

groups (?5d–i & 5j–m). Furthermore, in compounds 6h–k the

spacer between the carbohydrate moiety and the triazole ring

was elongated from methyl to ethyl allowing for a higher confor-

mational ﬂexibility (Scheme 2).

The mannosyl triazoles 5d–m and 6h–k were obtained by

reacting the known mannosyl alkynes 10,²⁹13³⁶and 14³⁷with

the azides 15d–m. Whereas the azides 15d–f are commercially

available, 15g,³⁸15h,³⁹15i,⁴⁰15j–l,⁴¹and 15m⁴⁰were obtained

by known procedures.

The cycloaddition of alkyne 14 and azides 15d–i under Cu(I)-

catalyzed click conditions^33,34yielded directly the anti-substituted

triazoles 5d–i in 27–77% (Table 1). However, due to the cumber-

some puriﬁcation of the unprotected mannosyl triazoles, test com-

pounds 5j–m were obtained by an alternative sequence starting

from the protected alkyne 10 and azides 15j–m followed by sapon-

iﬁcation of the intermediates 16j–m. The analogous cycloaddition

of butinyl mannoside 13 with azides 15h–k yielded the protected

triazoles 17h–k in 83–96%. Final deacetylation under Zemplén

conditions gave the test compounds 6h–k, which contain a linker

extended by an additional carbon between mannose and aglycone

(Table 1).

2.2. Synthesis of FimH antagonists modiﬁed at the anomeric

center

To avoid the low metabolic stability of O-mannosides like com-

pounds 5 and 6 due to potential cleavage by mannosidases, the

corresponding N-linked mannosyl triazoles 7 and C-mannosides 8

were prepared (Scheme 3). Mannosyl azide 18 was obtained

according to published procedures.⁴²The Cu(I)-catalyzed click

reaction of 18 with the commercially available acetylenes 19n–s

gave exclusively the anomerically pure anti-substituted a-D-man-

nosyl-triazoles 20n–s in 84–98% yield and after deacetylation the

test compounds 7n–s (Table 1).

Finally, the synthesis of triazolyl-methyl-C-mannosides 8n–s

(Scheme 3) started from mannosyl cyanide 21, which was obtained

6458

O. Schwardt et al. / Bioorg. Med. Chem. 19 (2011) 6454–6473

Table 1

Pharmacodynamic and pharmacokinetic parameters of mannosylated triazoles 5–8

(continued on next page)

O. Schwardt et al. / Bioorg. Med. Chem. 19 (2011) 6454–6473

6459

Table 1. (continued)

The IC₅₀s weredetermined witha cell-free competitive binding assay.⁴⁵Relative IC₅₀s (rIC₅₀) werecalculated bydividingthe IC₅₀ofthe substance ofinterest

by the IC₅₀of the reference compound 1b (entry 2). Passive permeation through an artiﬁcial membrane and retention therein was determined by PAMPA

(parallel artiﬁcial membrane permeation assay).⁵⁰Distribution coefﬁcients (logD_7.4values) were measured by a miniaturized shake ﬂask procedure.⁵¹

Thermodynamic solubility was measured by an equilibrium shake ﬂask approach.⁵²P_eeffective permeation; n.p. not permeable; n.d. not determined.

6460

O. Schwardt et al. / Bioorg. Med. Chem. 19 (2011) 6454–6473

OAc

OH

R

Ref.⁴²

b)

(19n-s)

O

AcO

HO

a)

OAc

N₃

N

9

18

20n-s

7n-s

N

R

Ref.⁴³

n: R = Ph

OAc

OH

OAc

o: R = 4-CH₃Ph

p: R = 3-Cl-Ph

q: R = 4-CF₃Ph

r: R = 3-pyr

OAc

O

c)

O

b)

O

AcO

HO

AcO

CN

NHBoc

s: R = CH₂OPh

21

22

23

OAc

OH

R

OAc

O

OH

O

d)

O

b)

(19n-s)

AcO

HO

a)

N

N₃

N

24

25n-s

8n-s

N

R

Scheme 3. Reagents and conditions: (a) CuSO₄ꢀ5H₂O, Na-ascorbate, t-BuOH/H₂O/THF (1:1:1), rt, 1–2 d (20n–s: 84–98%, 25n–s: 93–98%); (b) NaOMe, MeOH, rt, 3–6 h, (7n–s:

65–92%, 8n–s: 83–87%, 23: 95%); (c) H₂(4 bar), cat. Pd/C, Boc₂O, EtOAc, 1 d (72%); d) (i) concd HCl, dioxane/H₂O (2:1), 4 h; (ii) TfN₃, NaHCO₃, cat. CuSO₄ꢀ5H₂O, PhMe/H₂O/

MeOH, rt, 20 h; (iii) Ac₂O, pyridine, rt, 4 h (81%).

from 9 as reported earlier.⁴³Catalytic hydrogenation in the pres-

ence of Boc₂O (?22) followed by deacetylation led to the Boc-pro-

tected amine 23. Cleavage of the Boc-group, amine-azide

exchange⁴⁴and subsequent re-acetylation yielded azide 24 in

81% over three steps. The cycloaddition of 24 and the acetylenes

19n–s gave the anti-substituted triazoles 25n–s in excellent yields.

Final deprotection afforded the test compounds 8n–s (Table 1).

recognition domain (CRD) of FimH as previously reported.⁴⁵A sol-

uble recombinant protein consisting of the FimH-CRD (amino acid

residues 1–156), a C-terminal thrombin cleavage site and a 6His-

tag (FimH-CRD-Th-6His) was expressed in E. coli strain HM125

and puriﬁed by afﬁnity chromatography on a Ni-NTA column. For

the determination of IC₅₀values, a microtiter plate coated with

FimH-CRD-Th-6His was incubated with biotinylated Man

a(1-3)-

[Man (1-6)]-Manb(1-4)-GlcNAcb(1-4)-GlcNAcb-polyacrylamide

a

2.3. Biological evaluation

(TM-PAA) polymer conjugated to streptavidin-horseradish peroxi-

dase and the FimH antagonist in fourfold serial dilution (Fig. 4).

The assay was performed in duplicates and repeated twice for each

compound. To ensure comparability of different antagonists, the

For an initial biological in vitro characterization, a cell-free com-

petitive binding assay⁴⁵and later on, a cell-based aggregation as-

say⁴⁶were applied. Whereas for the cell-free competitive binding

assay only the CRD of the pili was expressed, the complete pili

are present in the cell-based assay format. Furthermore, both for-

mats are competitive assays, that is, the analyzed antagonists com-

pete with mannosides for the binding site. In the cell-free

competitive binding assay, the competitors are polymer-bound

trimannosides, whereas in the aggregation assay the antagonist

competes with more potent oligo- and polysaccharide chains¹⁴

present on the surface of erythrocytes.⁴⁷The interaction is further

complicated by the existence of a high- and a low-afﬁnity state of

the CRD of FimH. Aprikian et al. experimentally demonstrated that

in full-length ﬁmbriae the pilin domain stabilizes the CRD domain

in the low-afﬁnity state, whereas the CRD domain alone adopts the

high-afﬁnity state.⁴⁸Furthermore, it was recently shown that shear

stress can induce a conformational switch (twist in the b-sandwich

fold of the CRD domain) resulting in improved afﬁnity.⁴⁹Therefore,

different afﬁnities are expected when - as in the cell-based aggre-

gometry assay - full-length ﬁmbriae are present, when compared

to the CRD domain alone.

reference compound n-heptyl

a-D

-mannopyranoside (1b)^9,46was

tested in parallel on each individual microtiter plate. The afﬁnities

are reported relative to 1b as rIC₅₀in Table 1. The relative IC₅₀

(rIC₅₀) is the ratio of the IC₅₀of the test compound to the IC₅₀of

1b (entry 2).

Interestingly, all antagonists in Table 1 except methyl a-D-man-

noside (1a) exhibit nanomolar afﬁnities. When compared to 1a, an

up to 30-fold improvement was obtained. In the ﬁrst series, con-

taining a triazolyl-methyl moiety (5a–m, entries 3–15), 5j (entry

12) exhibits the highest afﬁnity with an IC₅₀of 70 nM. This is in

the range of n-heptyl

a-D-mannoside (1b), however, compared to

the biphenyl derivative 4b¹⁹(Fig. 1), this in fact represents a

18-fold reduction of afﬁnity (rIC₅₀0.06¹⁹for 4b vs. rIC₅₀1.1 for

5j). At this point, we should recollect that 4b and 5j address differ-

ent docking modes (out- and in-docking-mode) and therefore also

different structural environments.

Antagonists where the linker between the anomeric center and

the triazole is extended by an additional carbon (?6h–k, entries

16–19) show afﬁnities in the range of 200 nM and therefore - with

the exception of 4-pyridyl derivative 6j (entry 18) - two- to four-

fold higher afﬁnity compared to their counterparts with the shorter

linker. When the triazole is directly linked to the anomeric center

(?7n–s, entries 20–25) afﬁnities are 2- to 8-fold reduced, probably

as a consequence of the reduced ﬂexibility preventing an optimal

2.4. Cell-free competitive binding assay

The cell-free inhibition assay is based on the interaction of a

biotinylated polyacrylamide glycopolymer with the carbohydrate

O. Schwardt et al. / Bioorg. Med. Chem. 19 (2011) 6454–6473

6461

conformational switch has also been observed for

a

-CF₂-manno-

sides.⁵³As a consequence, the triazolyl-methyl group now is ori-

ented equatorially in C-mannoside 25n, while in 20n the triazole

moiety adopts an axial position.

Subsequent 2D-NOESY measurements (Fig. 5C and D) con-

ﬁrmed this analysis. For both compounds a sequence of seven

2D-NOESY experiments with increasing mixing times from 0.5 s

to 2.0 s in steps of 0.25 s was recorded. The intensity of the positive

signals grows with increasing mixing time and indicates the rela-

tive spatial proximity of a particular proton to that of the source

proton. The NOEs of the proton of interest (int_cross) were normal-

ized to the intensity of the diagonal peak of the source proton

(int_diag). Plotting these normalized intensities against the mixing

time results in a straight line for each pair of protons. The distances

r_ijwere then calculated from the slopes

r of the linear regression

according to r_ij= r_ref

(r_ref/r_ij)

^1/6, were r_refis the average distance

of the geminal protons H-6a and H-6b, which was chosen as refer-

ence (r_ref= 1.78 Å).^54,55

Typically, in the chair conformation of carbohydrates the vicinal

proton–proton distances are approx. 2.95 0.15 Å for a diaxial,

2.45 0.15 Å for an axial-equatorial and 2.50 0.20 Å for a diequa-

torial orientation.⁵⁶As shown in Figure 5A and B, the distances of

the ring protons in 20n and 25n determined from NOE experi-

ments correlate well with the theoretical values and support the

results obtained from the analysis of the coupling constants. In

summary, NMR spectroscopic data indicate that the mannopyran-

osyl chairs in these compounds adopt different conformations,

depending on the substituent at C-1.

This conformational analysis offers an explanation for the two-

fold reduction of afﬁnity found for most of the C-mannosides 8

compared to the corresponding N-linked triazoles 7. Due to the

inversion of the ring conformation in 8 (¹C₄vs ⁴C₁), the optimal

ﬁt into the hydrophilic mannose-binding pocket of FimH is

disturbed.

Figure 4. Examples of inhibition curves obtained from the cell-free competitive

binding assay.⁴⁵Each assay was run in duplicate and was repeated at least twice.

For antagonists 5h, 6i and the reference compound 1b IC₅₀values in the nM range

were obtained.

interaction of the aglycone with the tyrosine gate. Finally, the

C-mannosides 8n–s (entries 26–31), which do not exhibit the exo-

anomeric effect of the O-mannosides and therefore can more easily

adopt the optimal orientation within the tyrosine gate, surprisingly

show a twofold reduction in afﬁnity.

2.5. Aggregometry assay

The potential to disaggregate E. coli from guinea pig erythrocytes

(GPE) was determined for a variety of the mannosylated triazoles in

a function-based aggregometry assay.⁴⁶The measurements were

performed in triplicates and the corresponding IC₅₀values were cal-

culated by plotting the area under the curve (AUC) of disaggregation

2.7. Pharmacokinetic properties of mannosyl triazoles

Finally, the druglikeness of this new class of FimH antagonists

was addressed. For a successful po application in our UTI mouse

model,¹⁹FimH antagonists have to exhibit oral bioavailability,

metabolic stability and fast renal elimination to the urinary tract,

their place of action. For the evaluation of oral absorption and renal

excretion of the triazoles 5–8 physicochemical parameters such as

solubility, lipophilicity (distribution coefﬁcients, log D_7.4) and per-

meability were determined (Table 1). The mannosides of all four

against the concentration of the antagonists. n-Heptyl

pyranoside (1b) was again used as reference compound with an

IC₅₀of 77.1 M (Table 2, entry 1). While the antagonists 5e, 6j, 6k,

7o and 7q showed IC₅₀values in the range of 200–300 M, surpris-

ingly no activities could be determined for compounds 5j, 8q and 8r

up to a concentration of 700

M. As earlier observed,⁴⁶the activities

a-D-manno-

l

obtained from the aggregometry assay are approximately 1000-fold

lower than the afﬁnities determined in the target-based competi-

tive assay.

compound families (5–8) are all highly soluble (159 lg/mL to >

3 mg/mL) and therefore fulﬁll a ﬁrst prerequisite for absorption

in the gastrointestinal tract (GIT). All compounds showed low to

moderate log D_7.4values in the range of < -1.5 to 1.45. While these

parameters are beneﬁcial for renal excretion,⁵⁷oral absorption by

passive diffusion can only be expected to a minor extent. Indeed,

for none of the tested compounds a signiﬁcant permeation through

an artiﬁcial membrane (PAMPA,⁵⁰log P_e, P_e: effective permeation)

nor membrane retention could be detected. Whereas for a success-

ful oral absorption a log P_e>ꢁ5.7 and/or a membrane retention

%Mm > 80 % are required,⁵⁸the corresponding values for all tria-

zoles are far from being in this range. Overall, only poor absorption

from the GIT can be therefore expected.

2.6. Conformational analysis of mannosyl triazoles

Compared to their counterparts 7, where the triazole is directly

linked to the anomeric center, most of the C-mannosides 8 exhibit

a lower afﬁnity. By applying NMR techniques, it was investigated

whether this loss of afﬁnity originates from distorted ring conforma-

tions. Due to signal overlap, the unprotected mannosides 7 and 8

were not suited for the conformational analysis. However, for the

peracetylated derivatives 20n and 25n the ring conformation could

be assigned based on coupling constants and NOESY experiments.

First, the observed ³J coupling constants for their ring protons were

strikingly different. In 20n, they were in agreement with those ex-

pected for a regular ⁴C₁chair conformation of an

a-D-mannopyran-

3. Conclusions

osyl ring, with small J_1,2and J_2,3couplingsand large values for J_3,4and

J_4,5(Fig. 5A). In contrast, the large J_1,2coupling constant (8.4 Hz) and

small to medium values for J_2,3, J_3,4and J_4,5found for 25n, are in

Crystal structures indicate that the natural ligand oligoman-

nose-3¹⁰inserts into the tyrosine gate formed by Tyr48, Tyr137

and Ile52 of the carbohydrate recognition domain of FimH

(in-docking-mode). In contrast, the recently reported high-afﬁnity

agreement with a ring ﬂip of the

a-D-mannopyranosyl chair from

the common ⁴C₁to the unusual ¹C₄conformation (Fig. 5B). A similar

6462

O. Schwardt et al. / Bioorg. Med. Chem. 19 (2011) 6454–6473

Table 2

IC₅₀values of mannosylated triazoles determined in the aggregometry assay⁴⁶

The relative IC₅₀(rIC₅₀) was calculated by dividing the IC₅₀of the substance of interest by the IC₅₀of the

reference compound 1b. n.a., not active.

biphenyl mannoside 4c was shown to bind in the out-docking-

mode, that is, it establishes a -stacking interaction with Tyr48

tion, thus preventing an optimal ﬁt of the mannosyl moiety into

the hydrophilic mannose-binding pocket of FimH.

p-p

from the outside of the tyrosine gate.¹¹Based on docking studies,

we designed a series of low molecular weight mannosyl triazoles,

which exhibit an increased conformational ﬂexibility of the agly-

cone and therefore should allow for binding to the tyrosine gate

in the in-docking-mode. For their pharmacodynamical evaluation

two assay formats, a target-based binding assay⁴⁵and a function-

based aggregation assay,⁴⁶were applied. In general, all triazoles

5–8 showed nanomolar afﬁnities, but only one representative,

the 4-pyridyl derivative 5j, was as potent as the reference

compound n-heptyl mannoside (1b). Obviously, the high ﬂexibility

of the n-heptyl aglycone in 1b optimally fulﬁlls the spatial require-

ments of the tyrosine gate. In addition, the hydrophobic contacts

established by the substituted triazole aglycone within the tyro-

Finally, for a successful therapeutic application, FimH antago-

nists have to exhibit appropriate pharmacokinetic properties,

that is, oral bioavailability and fast renal elimination to the uri-

nary tract, their place of action. One prerequisite for absorption

in the GIT is sufﬁcient solubility, a property, which is fulﬁlled

by all synthesized antagonists. However, according to their lipo-

philicity and membrane permeation properties, the mannosyl tri-

azoles are not expected to be orally absorbed. Possible

improvements of the pharmacokinetic proﬁles of mannosyl tria-

zoles are currently studied.

4. Experimental part

4.1. Chemistry

sine gate in the in-docking-mode are less favorable than the

p-p-

stacking interaction of biphenyl derivatives^11,19with Tyr48 in the

out-docking-mode.

General. NMR spectra were recorded on a Bruker Avance

DMX-500 (500 MHz) spectrometer. Assignment of ¹H and ¹³C

NMR spectra was achieved using 2D methods (COSY, HSQC).

Chemical shifts are expressed in ppm using residual CHCl₃and

CD₂HOD as references. Optical rotations were measured using a

Perkin-Elmer Polarimeter 341. Electron spray ionization mass

Furthermore, the reduced afﬁnities of the triazolyl-methyl-C-

mannosides 8 can be rationalized by a disturbed interaction of

the mannose moiety. A conformational analysis by ¹H NMR and

NOESY NMR revealed that in contrast to the other three classes

of mannosyl triazoles (compounds 5, 6 and 7), the C-mannosides

8 do not adopt the common ⁴C₁but an unusual ¹C₄chair conforma-

O. Schwardt et al. / Bioorg. Med. Chem. 19 (2011) 6454–6473

6463

Figure 5. Coupling constants and proton-proton distances for peracetylated triazoles 20n (A) and 25n (B) determined by ¹H NMR and 2D-NOESY experiments; 2D-NOESY

spectra of 20n (C) and 25n (D) in CDCl₃with mixing times of 1.5 s (C) and 750 ms (D).

spectra (ESI-MS) were obtained on a Waters micromass ZQ. The

HRMS analyses were carried out using a Bruker QTOF. Reactions

were monitored by TLC using glass plates coated with silica gel

60 F₂₅₄(Merck) and visualized by using UV light and/or by charring

with a molybdate solution (a 0.02 M solution of ammonium cerium

sulfate dihydrate and ammonium molybdate tetrahydrate in aque-

ous 10% H₂SO₄). MPLC separations were carried out on a Combi-

Flash Companion from Teledyne Isco equipped with RediSep

normal-phase or C₁₈reversed-phase ﬂash columns. Tetrahydrofu-

rane (THF) was freshly distilled under argon over sodium and ben-

zophenone. Methanol (MeOH) was dried by reﬂuxing with sodium

methoxide and distilled immediately before use. Dichloromethane

(DCM), ethyl acetate (EtOAc), and toluene were dried by ﬁltration

over Al₂O₃(Fluka, type 5016 A basic).

(0.25 eq) and 1 M aq sodium ascorbate (0.5 eq) were added under

argon at rt. After stirring overnight the solvents were removed in

vacuo and the crude product was puriﬁed by MPLC on silica (petrol

ether/EtOAc) to yield 12a–c, 16j–m and 17h–k as colorless oils.

4.1.3. General procedure C for the synthesis of mannosyl

triazoles 20n–s and 25n–s

Azide 18⁴²or 24 (1.0 eq) and acetylene 19n–s (2.0 eq) were dis-

solved in THF/tert-BuOH/H₂O (1:1:1, 3 mL/0.1 mmol 18 or 24). The

mixture was degassed in an ultrasound bath under a ﬂow of argon

for 20 min. Then 0.2 M aq CuSO₄ꢀ5H₂O (0.2 eq) and 1 M aq sodium

ascorbate (0.4 eq) were added under argon at rt. After stirring for

1–2 d the solvents were removed in vacuo and the crude product

was puriﬁed by MPLC on silica (petrol ether/EtOAc) to yield 20n–

s and 25n–s as colorless oils.

4.1.1. General procedure A for the synthesis of mannosyl

triazoles 5d–i

4.1.4. General procedure D for deacetylation

A mixture of acetylene 14³⁷(1.0 eq), azide 15d–i (1.5 eq), Cu-

SO₄ꢀ5H₂O (0.25 eq) and sodium ascorbate (0.5 eq) was dissolved

in degassed tert-BuOH/H₂O (1:1, 2 mL/0.1 mmol 14) under argon.

After stirring for 1 d the solvents were removed in vacuo and the

crude product was ﬁrst puriﬁed by MPLC on silica (DCM/MeOH)

and then by reversed-phase chromatography (RP-18, H₂O/MeOH)

to yield 5d–i as colorless solids.

To a solution of the acetylated compound (38–50 mg) in MeOH

(3 mL) was added 1 M NaOMe/MeOH (0.3 mL). The mixture was

stirred at rt for 3–6 h. The solution was concentrated and the res-

idue was puriﬁed by MPLC on reversed phase (RP-18 column, H₂O/

MeOH) and P2 size-exclusion chromatography to afford the target

molecule as a colorless solid after a ﬁnal lyophilization from water/

dioxane.

4.1.2. General procedure B for the synthesis of mannosyl

triazoles 12a–c, 16j–m and 17h–k

4.1.5. Synthesis of azide 24

4.1.5.1.

(2,3,4,6-Tetra-O-acetyl-

a-D

-mannopyranosyl)-N-tert-

Cyanide 21⁴³(1.63 g,

Acetylene 10²⁹or 13³⁶(1.0 eq) and azide 11a–c or 15h–m

(1.5–2 eq) were dissolved in THF/tert-BuOH/H₂O (1:1:1, 1.5 mL/

0.1 mmol 10 or 11). The mixture was degassed in an ultrasound

bath under a ﬂow of argon for 20 min. Then 0.5 M aq CuSO₄ꢀ5H₂O

butoxycarbonyl-methylamine (22).

4.57 mmol), Boc₂O (1.49 g, 6.86 mmol) and Pd/C (10%, 250 mg)

were suspended in EtOAc (25 mL) and hydrogenated (4 bar H₂) at

rt for 4 h. After ﬁltration over Celite, fresh Pd/C (10%, 750 mg)

6464

O. Schwardt et al. / Bioorg. Med. Chem. 19 (2011) 6454–6473

was added and the mixture was hydrogenated (4 bar H₂) for addi-

tional 17 h. The suspension was ﬁltered over Celite and concen-

trated. The residue was puriﬁed by MPLC on silica (petrol ether/

EtOAc) to give 22 (1.51 g, 72%) as a colorless solid.

4.1.6. Synthesis of peracetylated mannosyl triazoles

4.1.6.1. Methyl 4-[4-((2,3,4,6-tetra-O-acetyl- -mannopyrano-

syloxy)methyl)-1H-1,2,3-triazol-1-yl]-benzoate (12a). Fol-

a-D

lowing general procedure B, 10 (40.0 mg, 0.103 mmol) was

¹H NMR (500 MHz, CDCl₃): d 1.44 (s, 9H, C(CH₃)₃), 2.07, 2.10,

2.10 (3 s, 12H, 4 COCH₃), 3.38 (m, 2H, H-1⁰), 3.99–4.05 (m, 2H, H-

1, H-5), 4.07 (dd, J = 3.8, 11.8 Hz, 1H, H-6a), 4.54 (dd, J = 6.9,

11.7 Hz, 1H, H-6b), 4.79 (m, 1H, NH), 5.07 (dd, J = 5.3, 6.4 Hz, 1H,

H-4), 5.10 (dd, J = 3.3, 6.0 Hz, 1H, H-2), 5.26 (dd, J = 3.3, 6.5 Hz,

1H, H-3); ¹³C NMR (125 MHz, CDCl₃): d 20.70, 20.73, 20.76, 20.81

(4 COCH₃), 28.3 (C(CH₃)₃), 39.7 (C-1⁰), 61.2 (C-6), 67.50, 67.51 (C-

2, C-4), 68.0 (C-3), 71.1 (C-1), 72.2 (C-5), 79.7 (C(CH₃)₃), 155.8

(NCO), 169.5, 169.6, 169.9, 170.7 (4 COCH₃); ESI-MS Calcd for

C₂₀H₃₁NNaO₁₁[M+Na]⁺: 484.18, Found: 484.11.

reacted with methyl 4-azidobenzoate (11a,³⁰36.5 mg,

0.206 mmol), 0.5 M CuSO₄(52

bate (52 L, 52 mol) to yield 12a (55.8 mg, 96%).

_D+45.0 (c 1.03, CHCl₃); IR (ﬁlm) 1747 (vs, CO) cm^ꢁ1; ¹H NMR

lL, 26 lmol) and 1 M sodium ascor-

l

[a]

(500 MHz, CDCl₃): d 1.99, 2.04, 2.12, 2.16 (4 s, 12H, 4 COCH₃), 3.97

(s, 3H, OMe), 4.10 (ddd, J = 2.2, 5.1, 9.5 Hz, 1H, H-5), 4.14 (dd,

J = 2.3, 12.2 Hz, 1H, H-6a), 4.32 (dd, J = 5.2, 12.2 Hz, 1H, H-6b),

4.79, 4.95 (A, B of AB, J = 12.6 Hz, 2H, H-1⁰), 5.01 (d, J = 1.1 Hz,

1H, H-1), 5.28 (dd, J = 1.7, 3.1 Hz, 1H, H-2), 5.32 (t, J = 9.8 Hz, 1H,

H-4), 5.36 (dd, J = 3.2, 10.0 Hz, 1H, H-3), 7.88 (AA⁰of AA⁰BB⁰,

J = 8.7 Hz, 2H, C₆H₄), 8.11 (s, 1H, C₂N₃H), 8.23 (BB⁰of AA⁰BB⁰,

J = 8.7 Hz, 2H, C₆H₄); ¹³C NMR (125 MHz, CDCl₃): d 20.7, 20.8,

20.9 (4C, 4 COCH₃), 52.5 (OMe), 61.0 (C-1⁰), 62.4 (C-6), 66.0 (C-4),

68.8, 69.0, 69.4 (C-2, C-3, C-5), 97.0 (C-1), 120.0 (2C, C₆H₄), 121.0

(C₂N₃H–C5), 130.4 (C₆H₄–C1), 131.4 (2C, C₆H₄), 139.9 (C₆H₄–C4),

144.7 (C₂N₃H–C4), 169.7, 170.0, 170.1, 170.7 (5C, 5 CO); ESI-MS

Calcd for C₂₅H₂₉N₃NaO₁₂[M+Na]⁺: 586.02, Found: 586.16.

4.1.5.2. N-tert-Butoxycarbonyl-(

a-D-mannopyranosyl)methyl-

amine (23). A solution of 22 (1.47 g, 3.19 mmol) in MeOH

(20 mL) was treated with 1 M methanolic NaOMe (2 mL) under ar-

gon at rt for 3 h. The reaction mixture was neutralized with acetic

acid and concentrated. The residue was puriﬁed by MPLC on silica

(DCM/MeOH) to give 23 (925 mg, 99%) as a colorless solid.

¹H NMR (500 MHz, CD₃OD): d 1.44 (s, 9H, C(CH₃)₃), 3.32 (m, 2H,

H-1⁰), 3.54 (m, 1H, H-5), 3.63 (t, J = 7.6 Hz, 1H, H-4), 3.68 (dd,

J = 3.1, 7.8 Hz, 1H, H-3), 3.74 (dd, J = 2.8, 11.8 Hz, 1H, H-6a), 3.77

(m, 1H, H-2), 3.79 (dd, J = 6.4, 11.8 Hz, 1H, H-6b), 3.86 (m, 1H, H-

4.1.6.2. Ethyl 3-[4-((2,3,4,6-tetra-O-acetyl-

a-

D-mannopyranosyl-

oxy)methyl)-1H-1,2,3-triazol-1-yl]-benzoate (12b).

Follow-

ing general procedure B, 10 (50.0 mg, 0.129 mmol) was reacted

with ethyl 3-azidobenzoate (11b,³¹49.3 mg, 0.258 mmol), 0.5 M

1), 6.72 (m, 1H, NH); ¹³C NMR (125 MHz, CD₃OD):

d 28.8

(C(CH₃)₃), 40.6 (C-1⁰), 62.7 (C-6), 69.6 (C-4), 70.2 (C-2), 72.7 (C-3),

76.9 (C-1), 77.4 (C-5), 80.2 (C(CH₃)₃), 158.6 (NCO); ESI-MS Calcd

for C₁₂H₂₃NNaO₇[M+Na]⁺: 316.14, Found: 316.03.

CuSO₄(64

64 mol) to yield 12b (72.7 mg, 97%).

_D+40.2 (c 1.04, CHCl₃); IR (ﬁlm) 1749 (vs, CO) cm^ꢁ1; ¹H NMR

lL, 32 lmol) and 1 M sodium ascorbate (64 lL,

l

[a]

(500 MHz, CDCl₃): d 1.43 (t, J = 7.2 Hz, 1H, CH₃), 1.99, 2.04, 2.12,

2.16 (4 s, 12H, 4 COCH₃), 4.09–4.15 (m, 2H, H-5, H-6a), 4.32 (dd,

J = 5.0, 12.1 Hz, 1H, H-6b), 4.44 (q, J = 7.2 Hz, 2H, OCH₂), 4.79,

4.95 (A, B of AB, J = 12.4 Hz, 2H, H-1⁰), 5.02 (d, J = 1.4 Hz, 1H, H-

1), 5.28 (dd, J = 1.7, 3.2 Hz, 1H, H-2), 5.31 (m, 1H, H-4), 5.36 (dd,

J = 3.3, 9.9 Hz, 1H, H-3), 7.64 (t, J = 8.0 Hz, 1H, C₆H₄–H5), 8.02

(ddd, J = 0.9, 2.1, 8.0 Hz, 1H, C₆H₄–H4), 8.11 (s, 1H, C₂N₃H), 8.14

(d, J = 7.9 Hz, 1H, C₆H₄-H6), 8.38 (t, J = 1.7 Hz, 1H, C₆H₄-H2); ¹³C

NMR (125 MHz, CDCl₃): d 14.3 (CH₃), 20.67, 20.69, 20.78, 20.88

(4 COCH₃), 60.9 (C-1⁰), 61.7 (OCH₂), 62.4 (C-6), 66.0 (C-4), 68.8

(C-3), 69.0 (C-5), 69.4 (C-2), 96.9 (C-1), 121.2 (C₆H₄), 121.3

(C₂N₃H–C5), 124.8, 129.9, 130.0, 132.3, 137.0 (C₆H₄), 144.5

(C₂N₃H–C4), 165.2, 169.7, 169.9, 170.1, 170.7 (5 CO); ESI-MS Calcd

for C₂₆H₃₂N₃O₁₂[M+H]⁺: 578.20, Found: 578.19.

4.1.5.3. (2,3,4,6-Tetra-O-acetyl-

ide (24).

a-D-mannopyranosyl)methylaz-

Triﬂyl azide stock solution preparation:⁴⁴Sodium

azide (796 mg, 12.2 mmol) was dissolved in water (2 mL). Toluene

(2 mL) was added, and the mixture was cooled to 0 °C with stirring.

Then triﬂic anhydride (1.31 mL, 6.12 mmol) was added dropwise.

The biphasic reaction mixture was stirred vigorously at 0 °C for

30 min and at 10 °C for another 2 h. The reaction mixture was neu-

tralized with satd aq NaHCO₃. The phases were separated, and the

aqueous phase extracted with toluene (2 ꢂ 2 mL). The organic

layers were combined to give the triﬂyl azide stock solution.

Amine-azide exchange: A solution of 23 (430 mg, 1.47 mmol) in

dioxane/water (2:1, 15 mL) was treated with concentrated HCl

(5 mL) under argon at rt for 4 h. The mixture was concentrated

and the residue was dried in high vacuo. Then, the crude amine

hydrochloride (341 mg), NaHCO₃(492 mg, 5.86 mmol) and Cu-

4.1.6.3. Methyl 5-[4-((2,3,4,6-tetra-O-acetyl-

syloxy)methyl)-1H-1,2,3-triazol-1-yl]-nicotinate (12c).

a

-

D

-mannopyrano-

Fol-

SO₄ꢀ5H₂O (14.1 mg, 61

lmol) were dissolved in water (1.91 mL).

The triﬂyl azide stock solution (3.25 mL, 3.3 mmol) was added

and the biphasic reaction mixture was made homogenous by the

addition of MeOH (12.6 mL). The mixture was stirred at rt for

20 h. The solvents were removed in vacuo and the residue was ta-

ken up in dry pyridine (10 mL), and acetic anhydride (4 mL) was

added. The reaction mixture was stirred at rt under argon for 4 h.

The solvents were removed in vacuo and the crude product was

puriﬁed by MPLC on silica (petrol ether/EtOAc) to yield 24

(459 mg, 81%) as a colorless oil.

lowing general procedure B, 10 (40.0 mg, 0.103 mmol) was

reacted with methyl 5-azidonicotinate (11c,³²32.5 mg,

0.182 mmol), 0.5 M CuSO₄(52

lL, 26 lmol) and 1 M sodium ascor-

bate (52 L, 52 mol). The crude product was dissolved in DCM

l

(10 mL) and washed with 0.1 M aq EDTA (5 mL). The aqueous

phase was extracted with DCM (2 ꢂ 10 mL) and the combined or-

ganic layers were dried with Na₂SO₄and evaporated to dryness.

The residue was puriﬁed by MPLC on silica (petrol ether/EtOAc)

to give 12c (42.4 mg, 73%).

IR (ﬁlm) 2102 (vs, N₃), 1747 (vs, CO) cm^ꢁ1

;

¹H NMR

[a]

_D+39.7 (c 1.06, CHCl₃); IR (ﬁlm) 1733 (vs, CO) cm^ꢁ1; ¹H NMR

(500 MHz, CDCl₃): d 2.04, 2.07, 2.08, 2.10 (4 s, 12H, 4 COCH₃),

3.29 (dd, J = 3.3, 13.4 Hz, 1H, H-1⁰a), 3.49 (dd, J = 7.3, 13.4 Hz,

1H, H-1⁰b), 4.05–4.09 (m, 2H, H-5, H-6a), 4.15 (dt, J = 3.2,

7.1 Hz, 1H, H-1), 4.60 (m, 1H, H-6b), 5.01 (dd, J = 4.4, 6.0 Hz,

1H, H-4), 5.14 (dd, J = 3.4, 6.9 Hz, 1H, H-2), 5.27 (dd, J = 3.4,

6.0 Hz, 1H, H-3); ¹³C NMR (125 MHz, CDCl₃): d 20.61, 20.63,

20.65, 20.74 (4 COCH₃), 50.1 (C-1⁰), 60.8 (C-6), 67.0 (C-2), 67.5

(C-3), 67.6 (C-4), 70.5 (C-1), 72.9 (C-5), 169.3, 169.5, 169.6,

(500 MHz, CDCl₃): d 1.98, 2.03, 2.11, 2.15 (4 s, 12H, 4 COCH₃), 4.01

(s, 3H, OCH₃), 4.09 (m, 1H, H-5), 4.14 (dd, J = 2.4, 12.2 Hz, 1H, H-6a),

4.31 (dd, J = 5.2, 12.3 Hz, 1H, H-6b), 4.79, 4.96 (A, B of AB,

J = 12.5 Hz, 2H, H-1⁰), 5.01 (d, J = 1.4 Hz, 1H, H-1), 5.27 (dd,

J = 1.7, 3.0 Hz, 1H, H-2), 5.30 (t, J = 9.8 Hz, 1H, H-4), 5.34 (dd,

J = 3.3, 9.9 Hz, 1H, H-3), 8.17 (s, 1H, C₂N₃H), 8.69 (t, J = 2.0 Hz, 1H,

C₅H₃N-H2), 9.27, 9.30 (2 s, 2H, C₅H₃N-H4, H6); ¹³C NMR

(125 MHz, CDCl₃): d 20.63, 20.65, 20.75, 20.84 (4 COCH₃), 52.9

(OMe), 60.8 (C-1⁰), 62.4 (C-6), 66.0 (C-4), 68.8, 68.9 (C-3, C-5),

69.3 (C-2), 97.0 (C-1), 121.2 (C₂N₃H–C5), 126.9, 128.6, 133.3

170.6

(4

COCH₃);

ESI-MS

Calcd

for

C₁₅H₂₁N₃NaO₉

[M+Na]⁺: 410.12, Found: 410.04.

O. Schwardt et al. / Bioorg. Med. Chem. 19 (2011) 6454–6473

6465

(C₅H₃N), 145.1 (2C, C₅H₃N, (C₂N₃H–C4), 150.7 (C₅H₃N), 164.4,

169.7, 169.9, 170.1, 170.7 (5 CO); ESI-MS Calcd for C₂₄H₂₉N₄O₁₂

[M+H]⁺: 565.18, Found: 565.15.

4.1.6.7. [1-(4⁰-Methoxyphenyl)-1,2,3-triazol-4-yl]methyl 2,3,4,6-

tetra-O-acetyl- -mannopyranoside (16m). Following

a-D

general procedure B, 10 (100 mg, 0.26 mmol) was reacted with

1-azido-4-methoxybenzene (15m,⁴⁰58 mg, 0.39 mmol), 0.5 M

4.1.6.4. [1-(Pyridin-4-yl)-1,2,3-triazol-4-yl]methyl 2,3,4,6-tetra-

CuSO₄(130

l

L, 65

mol) to yield 16m (128 mg, 92%).

_D+45.0 (c 2.26, DCM); IR (ﬁlm) 1749 (vs, CO) cm^ꢁ1

;

l

mol) and 1 M sodium ascorbate (130

lL,

O-acetyl-

procedure B, 10 (100 mg, 0.26 mmol) was reacted with 4-azido-

pyridine (15j,⁴¹47 mg, 0.39 mmol), 0.5 M CuSO₄(130

L, 65 mol)

and 1 M sodium ascorbate (130 L, 130 mol). The crude product

a

-D

-mannopyranoside (16j).

Following general

130

[

l

a

]

¹H NMR

l

(500 MHz, CDCl₃): d 1.93, 1.98, 2.06, 2.10 (4s, 12H, 4 COCH₃), 3.81

(s, 3H, OCH₃), 3.99–4.12 (m, 2H, H-5, H-6a), 4.26 (dd, J = 5.3,

12.4 Hz, 1H, H-6b), 4.71, 4.87 (A, B of AB, J = 12.4 Hz, 2H, H-1⁰),

4.95 (d, J = 1.4 Hz, 1H, H-1), 5.20–5.28 (m, 2H, H-2, H-4), 5.30

(dd, J = 3.2, 10.0 Hz, 1H, H-3), 6.97 (m, 2H, C₆H₄), 7.58 (m, 2H,

C₆H₄), 7.91 (s, 1H, C₂N₃H); ¹³C NMR (125 MHz, CDCl₃): d 20.78,

20.80, 20.89, 20.98 (4 COCH₃), 55.7 (OCH₃), 61.2 (C-1⁰), 62.5 (C-

6), 66.1, 68.9, 69.1, 69.5 (C-2, C-3, C-4, C-5), 97.0 (C-1), 114.9 (2C,

C₆H₄), 121.5 (C₂N₃H–C5), 122.4 (2C, C₆H₄), 130.4 (C₆H₄–C1),

144.1 (C₂N₃H–C4), 160.0 (C₆H₄–C4), 169.8, 170.0, 170.2, 170.8 (4

COCH₃); ESI-MS Calcd for C₂₄H₂₉N₃NaO₁₁[M+Na]⁺: 558.18, Found:

558.18.

l

was dissolved in DCM (20 mL) and washed with 0.1 M aq EDTA

(10 mL). The aqueous phase was extracted with DCM (2 ꢂ 20 mL)

and the combined organic layers were dried with Na₂SO₄and

evaporated to dryness. The residue was puriﬁed by MPLC on silica

(DCM/MeOH) to give 16j (114 mg, 87%).

[a] ;

_D+52.1 (c 2.26, DCM); IR (ﬁlm) 1748 (vs, CO) cm^{ꢁ1 1}H NMR

(500 MHz, CDCl₃): d 1.92, 1.97, 2.05, 2.09 (4s, 12H, 4 COCH₃), 3.96–

4.14 (m, 2H, H-5, H-6a), 4.24 (dd, J = 4.8, 12.1 Hz, 1H, H-6b), 4.73,

4.89 (A, B of AB, J = 12.5 Hz, 2H, H-1⁰), 4.94 (s, 1H, H-1), 5.14–

5.33 (m, 3H, H-2, H-3, H-4), 7.70 (m, 2H, C₅H₄N), 8.15 (s, 1H,

C₂N₃H), 8.73 (m, 2H, C₅H₄N); ¹³C NMR (125 MHz, CDCl₃): d

20.77, 20.78, 20.88, 20.96 (4 COCH₃), 61.0 (C-1⁰), 62.5 (C-6), 66.1,

68.9, 69.0, 69.5 (C-2, C-3, C-4, C-5), 97.1 (C-1), 113.9 (2C, C₅H₄N),

120.7 (C₂N₃H–C5), 143.0 (C₅H₄N–C1), 145.2 (C₂N₃H–C4), 151.9

(2C, C₅H₄N), 169.8, 170.1, 170.2, 170.8 (4 COCH₃); ESI-MS Calcd

for C₂₂H₂₆N₄NaO₁₀[M+Na]⁺: 529.16, Found: 529.07.

4.1.6.8. [1-(3⁰-Methoxybenzyl)-1,2,3-triazol-4-yl]ethyl 2,3,4,6-

tetra-O-acetyl-

a-

D-mannopyranoside

(17h).

Following

general procedure B, 13 (55 mg, 0.14 mmol) was reacted with 3-

methoxybenzylazide (15h,³⁹34 mg, 0.21 mmol), 0.5 M CuSO₄

(70 mol) to

yield 17h (65 mg, 83%).

;

_D+36.5 (c 1.00, DCM); IR (ﬁlm) 1748 (vs, CO) cm^{ꢁ1 1}H NMR

l

L, 35

l

mol) and 1 M sodium ascorbate (70

lL, 70

l

4.1.6.5. [1-(4⁰-Fluorophenyl)-1,2,3-triazol-4-yl]methyl 2,3,4,6-

[a

]

tetra-O-acetyl-

a

-

D

-mannopyranoside

(16k).

Following

(500 MHz, CDCl₃): d 1.85, 1.88, 1.93, 1.99 (4s, 12H, 4 COCH₃), 2.86

(tt, J = 4.8, 9.9 Hz, 2H, H-2⁰), 3.56 (dt, J = 6.6, 9.5 Hz, 1H, H-1⁰a), 3.63

(s, 3H, OCH₃), 3.69 (m, 1H, H-5), 3.83 (dt, J = 6.6, 9.5 Hz, 1H, H-1⁰b),

3.91 (dd, J = 2.4, 12.3 Hz, 1H, H-6a), 4.09 (dd, J = 5.2, 12.3 Hz, 1H, H-

6b), 4.65 (d, J = 1.7 Hz, 1H, H-1), 5.03 (dd, J = 1.8, 3.0 Hz, 1H, H-2),

5.07–5.16 (m, 2H, H-3, H-4), 5.33 (s, 2H, CH₂Ar), 6.68, 6.72, 7.13

(m, 4H, C₆H₄), 7.68 (s, 1H, C₂N₃H); ¹³C NMR (125 MHz, CDCl₃): d

20.85, 20.88, 21.02 (4C, 4 COCH₃), 26.4 (C-2⁰), 54.1 (CH₂Ar), 55.4

(OCH₃), 62.5 (C-6), 66.2 (C-4), 67.2 (C-1⁰), 68.9, 69.2, 69.7 (C-2, C-

3, C-5), 97.6 (C-1), 113.9, 114.2, 120.4, 130.3 (C₆H₄, C₂N₃H–C5),

136.5 (C₆H₄–C1), 144.8 (C₂N₃H–C4), 160.2 (C₆H₄–C3), 169.8,

170.1, 170.2, 170.8 (4 COCH₃); ESI-MS Calcd for C₂₆H₃₃N₃NaO₁₁

[M+Na]⁺: 586.21, Found 586.29.

general procedure B, 10 (100 mg, 0.26 mmol) was reacted with

1-azido-4-ﬂuorobenzene (15k,⁴¹53 mg, 0.39 mmol), 0.5 M CuSO₄

(130

to yield 16k (127 mg, 94%).

+42.0 (c 1.00, DCM); IR (ﬁlm) 1749 (vs, CO) cm^ꢁ1

;

lL, 65

l

mol) and 1 M sodium ascorbate (130

l

L, 130

lmol)

[

a]

D

¹H NMR

(500 MHz, CDCl₃): d 1.96, 2.01, 2.09, 2.13 (4s, 12H, 4 COCH₃), 4.07

(m, 1H, H-5), 4.11 (dd, J = 2.4, 12.2 Hz, 1H, H-6a), 4.28 (dd, J = 5.2,

12.2 Hz, 1H, H-6b), 4.75, 4.91 (A, B of AB, J = 12.4 Hz, 2H, H-1⁰),

4.98 (d, J = 1.6 Hz, 1H, H-1), 5.24–5.31 (m, 2H, H-2, H-4), 5.33

(dd, J = 3.3, 10.0 Hz, 1H, H-3), 7.21 (m, 2H, C₆H₄), 7.71 (m, 2H,

C₆H₄), 7.97 (s, 1H, C₂N₃H); ¹³C NMR (125 MHz, CDCl₃): d 20.87,

20.88, 20.98, 21.07 (4 COCH₃), 61.3 (C-1⁰), 62.6 (C-6), 66.3, 69.0,

69.2, 69.7 (C-2, C-3, C-4, C-5), 97.2 (C-1), 117.0 (d, J = 22.5 Hz, 2C,

C₆H₄), 121.6 (C₂N₃H–C5), 122.9 (d, J = 8.8 Hz, 2C, C₆H₄), 133.4 (d,

J = 3.8 Hz, C₆H₄–C1), 145.0 (C₂N₃H–C4), 163.8 (d, J = 247.5 Hz,

C₆H₄–C4), 169.9, 170.2, 170.3, 170.9 (4 COCH₃); ESI-MS Calcd for

4.1.6.9. [1-(4⁰-Nitrophenyl)-1,2,3-triazol-4-yl]ethyl 2,3,4,6-tetra-

O-acetyl-

procedure B, 13 (55 mg, 0.14 mmol) was reacted with 1-azido-4-

nitrobenzene (15i,⁴⁰34 mg, 0.21 mmol), 0.5 M CuSO₄(70

L,

35 mol) and 1 M sodium ascorbate (70 L, 70 mol) to yield 17i

(74 mg, 94%).

_D+28.1 (c 1.00, DCM); IR (ﬁlm) 1748 (vs, CO) cm^{ꢁ1 1}H NMR

a-

D-mannopyranoside (17i).

Following general

C

₂₃H₂₆FN₃NaO₁₀[M+Na]⁺: 546.16, Found: 546.15.

l

4.1.6.6. [1-(3⁰-Fluorophenyl)-1,2,3-triazol-4-yl]methyl 2,3,4,6-

tetra-O-acetyl- -mannopyranoside (16l). Following

a

-

D

[a

]

;

general procedure B, 10 (100 mg, 0.26 mmol) was reacted with

(500 MHz, CDCl₃): d 1.92, 1.99, 2.06, 2.11 (4s, 12H, 4 COCH₃), 3.12

(m, 2H, H-2⁰), 3.69–3.82 (m, 2H, H-5, H-1⁰a), 3.99–4.09 (m, 2H, H-

6a, H-1⁰b), 4.19 (dd, J = 5.3, 12.3 Hz, 1H, H-6b), 4.84 (d, J = 1.6 Hz,

1H, H-1), 5.17–5.27 (m, 2H, H-2, H-4), 5.30 (dd, J = 3.3, 10.2 Hz,

1H, H-3), 8.03 (s, 1H, C₂N₃H), 8.06, 8.37 (m, 4H, C₆H₄); ¹³C NMR

(125 MHz, CDCl₃): d 20.75, 20.89, 20.91, 21.02 (4 COCH₃), 26.3

(C-2⁰), 62.5 (C-6), 66.1 (C-4), 66.6 (C-1⁰), 69.1, 69.2, 69.6 (C-2, C-3,

C-5), 97.5 (C-1), 120.6 (C₂N₃H–C5), 120.6 (2C, C₆H₄), 125.6 (2C,

C₆H₄), 141.5 (C₆H₄–C1), 146.2 (C₂N₃H–C4), 147.2 (C₆H₄–C4),

169.7, 170.3, 170.4, 170.8 (4 COCH₃); ESI-MS Calcd for

1-azido-3-ﬂuorobenzene (15l,⁴¹53 mg, 0.39 mmol), 0.5 M CuSO₄

(130

to yield 16l (115 mg, 85%).

+47.5 (c 2.14, DCM); IR (ﬁlm) 1748 (vs, CO) cm^ꢁ1

l

L, 65

l

mol) and 1 M sodium ascorbate (130

l

L, 130

l

mol)

[

a]

D

;

¹H

NMR (500 MHz, CDCl₃): d 1.96, 2.01, 2.09, 2.13 (4s, 12H, 4

COCH₃), 4.02–4.15 (m, 2H, H-5, H-6a), 4.29 (dd, J = 5.1, 12.2 Hz,

1H, H-6b), 4.75, 4.91 (A, B of AB, J = 12.5 Hz, 2H, H-1⁰), 4.98 (s,

1H, H-1), 5.22–5.36 (m, 3H, H-2, H-3, H-4), 7.14 (m, 1H, C₆H₄),

7.44–7.59 (m, 3H, C₆H₄), 8.02 (s, 1H, C₂N₃H); ¹³C NMR

(125 MHz, CDCl₃): d 20.86, 20.88, 20.97, 21.07 (4 COCH₃), 61.2

(C-1⁰), 62.6 (C-6), 66.2, 69.0, 69.2, 69.6 (C-2, C-3, C-4, C-5),

97.2 (C-1), 108.6 (d, J = 26.3 Hz, C₆H₄), 116.0 (d, J = 17.5 Hz,

C₆H₄), 116.1 (m, C₆H₄), 121.3 (C₂N₃H–C5), 131.5 (d, J = 8.8 Hz,

C₆H₄), 138.2 (d, J = 10.0 Hz, C₆H₄–C1), 144.7 (C₂N₃H–C4), 163.3

(d, J = 247.5 Hz, C₆H₄–C4), 169.9, 170.1, 170.3, 170.9 (4 COCH₃);

C

₂₆H₃₃N₃NaO₁₁[M+Na]⁺: 587.17, Found 587.25.

4.1.6.10. [1-(Pyridin-4⁰-yl)-1,2,3-triazol-4-yl]ethyl 2,3,4,6-tetra-

O-acetyl- -mannopyranoside (17j). Following general

procedure B, 13 (60 mg, 0.15 mmol) was reacted with 4-azidopyri-

dine (15j,⁴¹28 mg, 0.23 mmol), 0.5 M CuSO₄(75

L, 38 mol) and

1 M sodium ascorbate (75 L, 75 mol). The crude product was

dissolved in DCM (20 mL) and washed with 0.1 M aq EDTA

a-D

l

ESI-MS Calcd for

546.15.

C

₂₃H₂₆FN₃NaO₁₀[M+Na]⁺: 546.16, Found:

l

6466

O. Schwardt et al. / Bioorg. Med. Chem. 19 (2011) 6454–6473

(10 mL). The aqueous phase was extracted with DCM (2 ꢂ 20 mL)

and the combined organic layers were dried with Na₂SO₄and

evaporated to dryness. The residue was puriﬁed by MPLC on silica

(petrol ether/EtOAc) to give 17j (74 mg, 94%).

J = 7.9 Hz, 4H, C₆H₄), 7.90 (s, 1H, C₂N₃H); ¹³C NMR (125 MHz,

CDCl₃): d 20.6, 20.7, 20.8 (4C, 4 COCH₃), 21.3 (PhCH₃), 61.6 (C-6),

66.1 (C-4), 68.3 (C-2), 68.8 (C-3), 72.1 (C-5), 83.6 (C-1), 119.3

(C₂N₃H–C5), 125.8, 126.8, 129.6, 138.6 (6C, C₆H₄), 148.4 (C₂N₃H–

C4), 169.3, 169.7, 169.7, 170.5 (4 COCH₃); ESI-MS Calcd for

[a] ;

_D+32.6 (c 0.99, DCM); IR (ﬁlm) 1748 (vs, CO) cm^{ꢁ1 1}H NMR

(500 MHz, CDCl₃): d 1.88, 1.97, 2.05, 2.10 (4s, 12H, 4 COCH₃), 3.10

(m, 2H, H-2⁰), 3.64–3.79 (m, 2H, H-5, H-1⁰a), 3.96–4.06 (m, 2H, H-

6a, H-1⁰b), 4.17 (dd, J = 5.3, 12.3 Hz, 1H, H-6b), 4.83 (d, J = 1.6 Hz,

1H, H-1), 5.16–5.26 (m, 2H, H-2, H-4), 5.28 (dd, J = 3.4, 10.1 Hz,

1H, H-3), 7.78 (dd, J = 1.6, 4.7 Hz, 2H, C₅H₄N), 8.04 (s, 1H, C₂N₃H),

8.72 (dd, J = 1.4, 4.9 Hz, 2H, C₅H₄N); ¹³C NMR (125 MHz, CDCl₃):

d 20.68, 20.88, 21.0 (4C, 4 COCH₃), 26.3 (C-2⁰), 62.5 (C-6), 66.0 (C-

4), 66.5 (C-1⁰), 69.0, 69.2, 69.7 (C-2, C-3, C-5), 97.4 (C-1), 113.9

(2C, C₅H₄N), 120.0 (C₂N₃H–C5), 143.3 (C₅H₄N–C1), 146.1 (C₂N₃H–

C4), 151.8 (2C, C₅H₄N), 169.7, 170.3, 170.4, 170.8 (4 COCH₃); ESI-

MS Calcd for C₂₆H₃₃N₃NaO₁₁[M+Na]⁺: 543.18, Found: 543.14.

C

₂₃H₂₇N₃NaO₉[M+Na]⁺: 512.16, Found: 512.15.

4.1.6.14. 1-(2,3,4,6-Tetra-O-acetyl-

a-

D-mannopyranosyl)-4-(3-

chlorophenyl)-1,2,3-triazole (20p).

Following general proce-

dure C, 18 (50 mg, 0.13 mmol) was reacted with 3-chloro-1-ethi-

nylbenzene (19p, 33 L, 0.27 mmol), 0.2 M CuSO₄(134 L,

27 mol) and 1 M sodium ascorbate (54 L, 54 mol) to yield

20p (59 mg, 86%).

+56.3 (c 1.03, CHCl₃); ¹H NMR (500 MHz, CDCl₃): d 2.07,

l

[a]

D

2.08, 2.09, 2.19 (4 s, 12H, 4 COCH₃), 3.95 (m, 1H, H-5), 4.08 (dd,

J = 2.1, 12.5 Hz, 1H, H-6a), 4.41 (dd, J = 5.4, 12.5 Hz, 1H, H-6b),

5.38 (t, J = 8.7 Hz, 1H, H-4), 5.95 (dd, J = 3.6, 8.6 Hz, 1H, H-3), 6.00

(m, 1H, H-2), 6.07 (d, J = 2.4 Hz, 1H, H-1), 7.34–7.40 (m, 2H,

C₆H₄–H5, H6), 7.75 (d, J = 7.4 Hz, 1H, C₆H₄–H4), 7.85 (s, 1H, C₆H₄-

H2), 7.98 (s, 1H, C₂N₃H); ¹³C NMR (125 MHz, CDCl₃): d 20.6, 20.7,

20.7, 20.8 (4 COCH₃), 61.5 (C-6), 66.1 (C-4), 68.2 (C-2), 68.7 (C-3),

72.4 (C-5), 83.5 (C-1), 120.1 (C₂N₃H–C5), 124.0 (C₆H₄–C4), 126.0

(C₆H₄–C2), 128.7, 130.3 (C₆H₄–C5, C6), 131.5 (C₆H₄–C3), 134.9

(C₆H₄–C1), 147.2 (C₂N₃H–C4), 169.3, 169.6, 169.7, 170.5 (4

COCH₃); ESI-MS Calcd for C₂₂H₂₄ClN₃NaO₉[M+Na]⁺: 532.11,

Found: 532.13.

4.1.6.11. [1-(4⁰-Fluorophenyl)-1,2,3-triazol-4-yl]ethyl 2,3,4,6-

tetra-O-acetyl-

a-

D-mannopyranoside (17k).

Following gen-

eral procedure B, 13 (60 mg, 0.15 mmol) was reacted with 1-azi-

do-4-ﬂuorobenzene (15k,⁴¹32 mg, 0.23 mmol), 0.5 M CuSO₄

(77 mol) to

yield 17k (77 mg, 96%).

;

_D+32.0 (c 1.01, DCM); IR (ﬁlm) 1751 (vs, CO) cm^{ꢁ1 1}H NMR

lL, 38

lmol) and 1 M sodium ascorbate (75

lL, 75

l

[a

]

(500 MHz, CDCl₃): d 1.90, 1.96, 2.05, 2.10 (4s, 12H, 4 COCH₃), 3.07

(m, 2H, H-2⁰), 3.67–3.79 (m, 2H, H-5, H-1⁰a), 3.96–4.06 (m, 2H, H-

6a, H-1⁰b), 4.18 (dd, J = 5.2, 12.3 Hz, 1H, H-6b), 4.82 (d, J = 1.6 Hz,

1H, H-1), 5.16–5.24 (m, 2H, H-2, H-4), 5.28 (dd, J = 3.4, 10.1 Hz,

1H, H-3), 7.16, 7.73 (m, 4H, C₆H₄), 7.85 (s, 1H, C₂N₃H); ¹³C NMR

(125 MHz, CDCl₃): d 20.71, 20.85, 20.88, 21.01 (4 COCH₃), 26.3

(C-2⁰), 62.5 (C-6), 66.1 (C-4), 66.8 (C-1⁰), 68.9, 69.2, 69.7 (C-2, C-3,

C-5), 97.4 (C-1), 116.8 (d, J = 22.5 Hz, 2C, C₆H₄), 120.7 (C₂N₃H–

C5), 122.6 (d, J = 8.8 Hz, 2C, C₆H₄), 133.6 (d, J = 2.5 Hz, C₆H₄–C1),

145.4 (C₂N₃H–C4), 162.5 (d, J = 247.5 Hz, C₆H₄–C4), 169.7, 170.2,

170.3, 170.8 (4 COCH₃); ESI-MS Calcd for C₂₆H₃₃N₃NaO₁₁

[M+Na]⁺: 560.18, Found 560.17.

4.1.6.15. 1-(2,3,4,6-Tetra-O-acetyl-

triﬂuoromethylphenyl)-1,2,3-triazole (20q).

eral procedure C, 18 (50 mg, 0.13 mmol) was reacted with 1-ethi-

nyl-4-triﬂuoromethylbenzene (19q, 44 L, 0.27 mmol), 0.2 M

CuSO₄(134 L, 27 mol) and 1 M sodium ascorbate (54 L,

54 mol) to yield 20q (62 mg, 85%).

+54.5 (c 0.95, CHCl₃); ¹H NMR (500 MHz, CDCl₃): d 2.08,

a-

D

-mannopyranosyl)-4-(4-

Following gen-

l

[a]

D

2.09, 2.19 (3 s, 12H, 4 COCH₃), 3.97 (ddd, J = 2.5, 5.4, 8.7 Hz, 1H,

H-5), 4.09 (dd, J = 2.5, 12.5 Hz, 1H, H-6a), 4.41 (dd, J = 5.5,

12.5 Hz, 1H, H-6b), 5.39 (t, J = 8.8 Hz, 1H, H-4), 5.95 (dd, J = 3.7,

8.7 Hz, 1H, H-3), 6.00 (dd, J = 3.1, 3.4 Hz, 1H, H-2), 6.09 (d,

J = 2.8 Hz, 1H, H-1), 7.71, 7.98 (AA⁰, BB⁰of AA⁰BB⁰, J = 8.1 Hz, 4H,

C₆H₄), 8.05 (s, 1H, C₂N₃H); ¹³C NMR (125 MHz, CDCl₃): d 20.60,

20.69, 20.70, 20.73 (4 COCH₃), 61.4 (C-6), 66.0 (C-4), 68.2 (C-2),

68.7 (C-3), 72.5 (C-5), 83.5 (C-1), 120.6 (C₂N₃H–C5), 124.0 (q,

J = 272 Hz, CF₃), 126.0 (q, J = 3.8 Hz, 2C, C₆H₄–C3, C5), 126.1 (2C,

C₆H₄–C2, C6), 130.5 (d, J = 32.6 Hz, C₆H₄–C4), 133.1 (C₆H₄–C1),

147.0 (C₂N₃H–C4), 169.3, 169.6, 169.7, 170.5 (4 COCH₃); ESI-MS

Calcd for C₂₃H₂₄F₃N₃NaO₉[M+Na]⁺: 566.14, Found: 566.10.

4.1.6.12. 1-(2,3,4,6-Tetra-O-acetyl-

nyl-1,2,3-triazole (20n). Following general procedure C, 18

(102 mg, 0.273 mmol) was reacted with phenylacetylene (19n,

60 L, 0.55 mmol), 0.2 M CuSO₄(273 L, 54.6 mol) and 1 M so-

dium ascorbate (109 L, 109 mol) to yield 20n (110 mg, 84%).

+65.5 (c 1.01, CHCl₃); ¹H NMR (500 MHz, CDCl₃): d 2.07,

a-D-mannopyranosyl)-4-phe-

l

[a]

D

2.07, 2.09, 2.20 (4 s, 12H, 4 COCH₃), 3.94 (ddd, J = 2.5, 5.4, 9.0 Hz,

1H, H-5), 4.08 (dd, J = 2.5, 12.5 Hz, 1H, H-6a), 4.39 (dd, J = 5.4,

12.5 Hz, 1H, H-6b), 5.39 (t, J = 8.9 Hz, 1H, H-4), 5.98 (dd, J = 3.7,

8.8 Hz, 1H, H-3), 6.02 (dd, J = 2.7, 3.7 Hz, 1H, H-2), 6.07 (d,

J = 2.6 Hz, 1H, H-1), 7.32–7.39, 7.44–7.47, 7.85–7.87 (m, 5H,

C₆H₅), 7.96 (s, 1H, C₂N₃H); ¹³C NMR (125 MHz, CDCl₃): d 20.5,

20.6, 20.6, 20.7 (4 COCH₃), 61.5 (C-6), 66.0 (C-4), 68.2 (C-2), 68.7

(C-3), 72.1 (C-5), 83.5 (C-1), 119.7 (C₂N₃H–C5), 125.8, 128.6,

128.9, 129.6 (6C, C₆H₅), 148.2 (C₂N₃H–C4), 169.2, 169.6, 169.6,

170.4 (4 COCH₃); ESI-MS Calcd for C₂₂H₂₅N₃NaO₉[M+Na]⁺:

498.15, Found: 498.20.

4.1.6.16. 1-(2,3,4,6-Tetra-O-acetyl-

pyridyl)-1,2,3-triazole (20r). Following general procedure C,

18 (50 mg, 0.13 mmol) was reacted with 3-ethinylpyridine (19r,

27.6 mg, 0.27 mmol), 0.2 M CuSO₄(134 L, 27 mol) and 1 M so-

dium ascorbate (54 L, 54 mol). Then the reaction mixture was

a-D-mannopyranosyl)-4-(3-

l

diluted with DCM (20 mL) and extracted with 25 mM aq EDTA

(10 mL). The organic layer was dried (Na₂SO₄), concentrated and

the residue was puriﬁed by MPLC on silica (petrol ether/EtOAc)

to yield 20r (61 mg, 96%).

4.1.6.13. 1-(2,3,4,6-Tetra-O-acetyl-

a-

D

-mannopyranosyl)-4-(4-

methylphenyl)-1,2,3-triazole (20o).

Following general pro-

[a]

+56.0 (c 0.70, CHCl₃); ¹H NMR (500 MHz, CDCl₃): d 2.06,

D

cedure C, 18 (50 mg, 0.13 mmol) was reacted with p-tolylacetylene

(19o, 34 L, 0.27 mmol), 0.2 M CuSO₄(134 L, 27 mol) and 1 M

sodium ascorbate (54 L, 54 mol) to yield 20o (64 mg, 98%).

+62.4 (c 1.09, CHCl₃); ¹H NMR (500 MHz, CDCl₃): d 2.06,

2.07, 2.09, 2.19 (4 s, 12H, 4 COCH₃), 2.39 (s, 3H, PhCH₃), 3.94

(ddd, J = 2.4, 5.3, 9.0 Hz, 1H, H-5), 4.07 (dd, J = 2.4, 12.5 Hz, 1H,

H-6a), 4.38 (dd, J = 5.4, 12.5 Hz, 1H, H-6b), 5.39 (t, J = 8.9 Hz, 1H,

H-4), 5.98 (dd, J = 3.7, 8.7 Hz, 1H, H-3), 6.01 (dd, J = 2.6, 3.7 Hz,

1H, H-2), 6.05 (d, J = 2.4 Hz, 1H, H-1), 7.26, 7.74 (AA⁰, BB⁰of AA⁰BB⁰,

2.06, 2.07, 2.17 (4 s, 12H, 4 COCH₃), 3.95 (ddd, J = 2.6, 5.5, 8.8 Hz,

1H, H-5), 4.07 (dd, J = 2.6, 12.5 Hz, 1H, H-6a), 4.40 (dd, J = 5.6,

12.5 Hz, 1H, H-6b), 5.38 (t, J = 8.8 Hz, 1H, H-4), 5.92 (dd, J = 3.7,

8.8 Hz, 1H, H-3), 6.00 (dd, J = 3.2, 3.5 Hz, 1H, H-2), 6.11 (d,

J = 2.9 Hz, 1H, H-1), 7.45 (dd, J = 4.9, 7.2 Hz, 1H, C₅H₄N-H5), 8.11

(s, 1H, C₂N₃H), 8.28 (d, J = 7.9 Hz, 1H, C₅H₄N-H6), 8.62 (m, 1H,

C₅H₄N-H4), 9.06 (br s, 1H, C₅H₄N–H2); ¹³C NMR (125 MHz, CDCl₃):

d 20.56, 20.66, 20.68, 20.70 (4 COCH₃), 61.4 (C-6), 66.0 (C-4), 68.1

(C-2), 68.6 (C-3), 72.5 (C-5), 83.6 (C-1), 120.4 (C₂N₃H–C5), 124.1

l

[a]

D

O. Schwardt et al. / Bioorg. Med. Chem. 19 (2011) 6454–6473

6467

(C₅H₄N–C5), 126.3 (C₅H₄N–C1), 133.7 (C₅H₄N–C6), 145.0 (C₂N₃H–

ethinylbenzene (19p, 25

21 mol) and 1 M sodium ascorbate (41

25p (51 mg, 94%).

-2.55 (c 1.27, CHCl₃); ¹H NMR (500 MHz, CDCl₃): d 1.84,

l

L, 0.21 mmol), 0.2 M CuSO₄(103

lL,

C4), 146.5 (C₅H₄N–C2), 148.9 (C₅H₄N–C4), 169.3, 169.6, 169.6,

170.5 (4 COCH₃); ESI-MS Calcd for C₂₁H₂₄N₄NaO₉[M+Na]⁺:

477.16, Found: 477.08.

l

L, 41 mol) to yield

l

[a]

D

2.07, 2.10, 2.11 (4 s, 12H, 4 COCH₃), 3.99 (dd, J = 4.0, 12.2 Hz, 1H,

H-6a), 4.15 (dt, J = 3.4, 9.0 Hz, 1H, H-5), 4.38 (dt, J = 2.7, 8.6 Hz,

1H, H-1), 4.51 (dd, J = 8.7, 14.4 Hz, 1H, H-1⁰a), 4.60 (dd, J = 9.2,

12.2 Hz, 1H, H-6b), 4.65 (dd, J = 2.7, 14.4 Hz, 1H, H-1⁰b), 4.93 (dd,

J = 2.9, 4.9 Hz, 1H, H-4), 5.03 (dd, J = 3.3, 8.5 Hz, 1H, H-2), 5.34

(dd, J = 3.6, 4.6 Hz, 1H, H-3), 7.27 (m, 1H, C₆H₄-H6), 7.33 (t,

J = 7.8 Hz, 1H, C₆H₄–H5), 7.72 (d, J = 7.7 Hz, 1H, C₆H₄–H4), 7.81 (t,

J = 1.7 Hz, 1H, C₆H₄-H2), 7.94 (s, 1H, C₂N₃H); ¹³C NMR (125 MHz,

CDCl₃): d 20.41, 20.60, 20.64, 20.73 (4 COCH₃), 50.4 (C-1⁰), 59.9

(C-6), 66.7 (C-2), 66.8 (C-3), 68.0 (C-4), 68.3 (C-1), 73.5 (C-5),

121.3 (C₂N₃H–C5), 123.6, 125.6, 128.1, 130.1, 132.3, 134.8 (C₆H₄),

146.5 (C₂N₃H–C4), 169.1, 169.3, 169.5, 170.3 (4 COCH₃); ESI-MS

Calcd for C₂₃H₂₇ClN₃O₉[M+H]⁺: 524.14, Found: 524.04.

4.1.6.17.

phenoxymethyl-1,2,3-triazole (20s).

cedure C, 18 (50 mg, 0.13 mmol) was reacted with phenylpropargyl

ether (19s, 34 L, 0.27 mmol), 0.2 M CuSO₄(134 L, 27 mol) and

1 M sodium ascorbate (54 L, 54 mol) to yield 20s (58 mg, 85%).

+38.8 (c 0.61, CHCl₃); ¹H NMR (500 MHz, CDCl₃): d 2.06,

2.07, 2.09, 2.18 (4 s, 12H, 4 COCH₃), 3.90 (m, 1H, H-5), 4.05 (dd,

J = 2.0, 12.5 Hz, 1H, H-6a), 4.37 (dd, J = 5.4, 12.5 Hz, 1H, H-6b),

5.26 (s, 2H, CH₂OPh), 5.37 (t, J = 8.9 Hz, 1H, H-4), 5.93 (dd, J = 3.6,

8.8 Hz, 1H, H-3), 5.97 (dd, J = 2.7, 3.3 Hz, 1H, H-2), 6.00 (d,

J = 2.3 Hz, 1H, H-1), 6.98–7.00, 7.30–7.32 (m, 5H, C₆H₅), 7.81 (s,

1H, C₂N₃H); ¹³C NMR (125 MHz, CDCl₃): d 20.59, 20.67, 20.69,

20.73 (4 COCH₃), 61.5 (C-6), 61.8 (CH₂OPh), 66.0 (C-4), 68.2 (C-2),

68.7 (C-3), 72.2 (C-5), 83.6 (C-1), 114.7 (2C, C₆H₅–C2, C6), 121.4

(C₆H₅–C4), 123.0 (C₂N₃H–C5), 129.6 (2C, C₆H₅–C3, C5), 145.2

(C₂N₃H–C4), 158.0 (C₆H₅–C1), 169.3, 169.6, 169.7, 170.5 (4 COCH₃);

ESI-MS Calcd for C₂₃H₂₇N₃NaO₁₀[M+Na]⁺: 528.16, Found: 528.14.

1-(2,3,4,6-Tetra-O-acetyl-

a

-

D

-mannopyranosyl)-4-

Following general pro-

l

[a]

D

4.1.6.21. 1-(2,3,4,6-Tetra-O-acetyl-

4-(4-triﬂuoromethylphenyl)-1,2,3-triazole (25q).

general procedure C, 24 (40 mg, 0.10 mmol) was reacted with 1-

ethinyl-4-triﬂuoromethylbenzene (19q, 34 L, 0.21 mmol), 0.2 M

CuSO₄(103 L, 21 mol) and 1 M sodium ascorbate (41 L,

41 mol) to yield 25q (56 mg, 98%).

]_D+0.47 (c 1.19, CHCl₃); ¹H NMR (500 MHz, CDCl₃): d 1.82, 2.08,

a

-D

-mannopyranosyl)methyl-

Following

l

4.1.6.18. 1-(2,3,4,6-Tetra-O-acetyl-

a-

D-mannopyranosyl)methyl-

l

[a

4-phenyl-1,2,3-triazole (25n).

Following general procedure

C, 24 (40 mg, 0.10 mmol) was reacted with phenylacetylene

(19n, 23 L, 0.21 mmol), 0.2 M CuSO₄(103 L, 21 mol) and 1 M

sodium ascorbate (41 L, 41 mol) to yield 25n (47 mg, 93%).

-1.76 (c 1.50, CHCl₃); ¹H NMR (500 MHz, CDCl₃): d 1.83,

2.10, 2.12 (4 s, 12H, 4 COCH₃), 3.97 (dd, J = 4.0, 12.2 Hz, 1H, H-6a),

4.15 (dt, J = 3.4, 9.0 Hz, 1H, H-5), 4.40 (dt, J = 2.5, 8.6 Hz, 1H, H-1),

4.53 (dd, J = 8.7, 14.4 Hz, 1H, H-1⁰a), 4.63 (dd, J = 9.2, 12.2 Hz, 1H,

H-6b), 4.67 (dd, J = 2.7, 14.4 Hz, 1H, H-1⁰b), 4.93 (dd, J = 2.9, 4.7 Hz,

1H, H-4), 5.04 (dd, J = 3.2, 8.6 Hz, 1H, H-2), 5.34 (t, J = 4.0 Hz, 1H,

H-3), 7.66, 7.95 (AA⁰, BB⁰of AA⁰BB⁰, J = 8.2 Hz, 4H, C₆H₄), 8.00 (s,

1H, C₂N₃H); ¹³C NMR (125 MHz, CDCl₃): d 20.42, 20.63, 20.67,

20.76 (4 COCH₃), 50.4 (C-1⁰), 59.9 (C-6), 66.7 (C-2), 66.8 (C-3), 68.0

(C-4), 68.2 (C-1), 73.5 (C-5), 121.7 (C₂N₃H–C5), 124.0 (q,

J = 272 Hz, CF₃), 125.7 (2C, C₆H₄–C2, C6), 125.8 (q, J = 3.8 Hz, 2C,

C₆H₄–C3, C5), 129.9 (q, J = 32.5 Hz, C₆H₄–C4), 133.9 (C₆H₄–C1),

146.4 (C₂N₃H–C4), 169.1, 169.4, 169.6, 170.3 (4 COCH₃); ESI-MS

Calcd for C₂₄H₂₇F₃N₃O₉[M+H]⁺: 558.17, Found: 558.22.

l

[a]

D

2.08, 2.11, 2.12 (4 s, 12H, 4 COCH₃), 4.00 (dd, J = 3.9, 12.2 Hz, 1H,

H-6a), 4.16 (dt, J = 3.5, 9.0 Hz, 1H, H-5), 4.39 (dt, J = 2.6, 8.6 Hz,

1H, H-1), 4.50 (dd, J = 8.8, 14.3 Hz, 1H, H-1⁰a), 4.59 (dd, J = 9.2,

12.1 Hz, 1H, H-6b), 4.66 (dd, J = 2.4, 14.3 Hz, 1H, H-1⁰b), 4.94 (dd,

J = 3.0, 4.9 Hz, 1H, H-4), 5.06 (dd, J = 3.2, 8.4 Hz, 1H, H-2), 5.35 (t,

J = 4.0 Hz, 1H, H-3), 7.32 (t, J = 7.4 Hz, 1H, C₆H₅-H4), 7.41 (t,

J = 7.4 Hz, 2H, C₆H₅-H3, H5), 7.83 (d, J = 8.0 Hz, 2H, C₆H₅-H2, H6),

7.92 (s, 1H, C₂N₃H); ¹³C NMR (125 MHz, CDCl₃): d 20.4, 20.6,

20.7, 20.8 (4 COCH₃), 50.3 (C-1⁰), 60.0 (C-6), 66.8 (C-2), 66.9 (C-

3), 68.0 (C-4), 68.4 (C-1), 73.4 (C-5), 120.9 (C₂N₃H–C5), 125.6,

128.2, 128.8, 130.5 (6C, C₆H₅), 148.0 (C₂N₃H–C4), 169.1, 169.4,

169.6, 170.4 (4 COCH₃); ESI-MS Calcd for C₂₃H₂₈N₃O₉[M+H]⁺:

490.18, Found: 490.17.

4.1.6.22. 1-(2,3,4,6-Tetra-O-acetyl-

a

-D

-mannopyranosyl)methyl-

4-(3-pyridyl)-1,2,3-triazole (25r).

Following general proce-

dure C, 24 (40 mg, 0.10 mmol) was reacted with 3-ethinylpyridine

(19r, 21.2 mg, 0.206 mmol), 0.2 M CuSO₄(103 L, 21 mol) and

1 M sodium ascorbate (41 L, 41 mol) to yield 25r (50 mg, 98%).

-0.08 (c 1.04, CHCl₃); ¹H NMR (500 MHz, CDCl₃): d 1.83,

l

4.1.6.19. 1-(2,3,4,6-Tetra-O-acetyl-

4-(4-methylphenyl)-1,2,3-triazole (25o).

procedure C, 24 (40 mg, 0.10 mmol) was reacted with p-tolylacet-

ylene (19o, 26 L, 0.21 mmol), 0.2 M CuSO₄(103 L, 21 mol) and

1 M sodium ascorbate (41 L, 41 mol) to yield 25o (50 mg, 97%).

-1.57 (c 1.26, CHCl₃); ¹H NMR (500 MHz, CDCl₃): d 1.83,

a-

D

-mannopyranosyl)methyl-

l

Following general

[a]

D

2.07, 2.09, 2.10 (4 s, 12H, 4 COCH₃), 3.97 (dd, J = 4.0, 12.2 Hz, 1H,

H-6a), 4.14 (dt, J = 3.4, 8.9 Hz, 1H, H-5), 4.39 (dt, J = 2.6, 8.6 Hz,

1H, H-1), 4.54 (dd, J = 8.6, 14.4 Hz, 1H, H-1⁰a), 4.60 (dd, J = 9.1,

12.2 Hz, 1H, H-6b), 4.66 (dd, J = 2.7, 14.4 Hz, 1H, H-1⁰b), 4.92 (dd,

J = 2.9, 4.8 Hz, 1H, H-4), 5.02 (dd, J = 3.3, 8.5 Hz, 1H, H-2), 5.33

(m, 1H, H-3), 7.37 (m, 1H, C₅H₄N-H5), 8.02 (s, 1H, C₂N₃H), 8.21

(d, J = 7.9 Hz, 1H, C₅H₄N-H6), 8.56 (s, 1H, C₅H₄N–H2), 9.00 (m,

1H, C₅H₄N-H4); ¹³C NMR (125 MHz, CDCl₃): d 20.44, 20.61, 20.65,

20.74 (4 COCH₃), 50.4 (C-1⁰), 59.9 (C-6), 66.6 (C-2), 66.8 (C-3),

67.9 (C-4), 68.3 (C-1), 73.5 (C-5), 121.3 (C₂N₃H–C5), 123.8

(C₅H₄N–C5), 126.8 (C₅H₄N–C1), 133.0 (C₅H₄N–C6), 144.6

(C₂N₃H–C4), 146.7 (C₅H₄N–C2), 149.0 (C₅H₄N–C4), 169.1, 169.3,

169.5, 170.3 (4 COCH₃); ESI-MS Calcd for C₂₂H₂₇N₄O₉[M+H]⁺:

491.18, Found: 491.17.

l

[a]

D

2.07, 2.10, 2.11 (4 s, 12H, 4 COCH₃), 2.36 (s, 3H, PhCH₃), 3.99 (dd,

J = 4.0, 12.2 Hz, 1H, H-6a), 4.15 (dt, J = 3.5, 9.0 Hz, 1H, H-5), 4.38

(dt, J = 2.7, 8.6 Hz, 1H, H-1), 4.50 (dd, J = 8.8, 14.4 Hz, 1H, H-1⁰a),

4.58 (dd, J = 9.1, 12.2 Hz, 1H, H-6b), 4.64 (dd, J = 2.8, 14.4 Hz, 1H,

H-1⁰b), 4.94 (dd, J = 3.1, 5.0 Hz, 1H, H-4), 5.06 (dd, J = 3.3, 8.3 Hz,

1H, H-2), 5.34 (dd, J = 3.5, 4.8 Hz, 1H, H-3), 7.21, 7.70 (AA⁰, BB⁰of

AA⁰BB⁰, J = 8.0 Hz, 4H, C₆H₄), 7.87 (s, 1H, C₂N₃H); ¹³C NMR

(125 MHz, CDCl₃): d 20.41, 20.62, 20.64, 20.74 (4 COCH₃), 21.2

(PhCH₃), 50.2 (C-1⁰), 60.0 (C-6), 66.8 (C-2), 66.9 (C-3), 68.0 (C-4),

68.5 (C-1), 73.3 (C-5), 120.5 (C₂N₃H–C5), 125.5, 127.6, 129.5,

138.0 (6C, C₆H₄), 147.8 (C₂N₃H–C4), 169.1, 169.4, 169.6, 170.4 (4

COCH₃); ESI-MS Calcd for C₂₄H₃₀N₃O₉[M+H]⁺: 504.20, Found:

504.20.

4.1.6.23. 1-(2,3,4,6-Tetra-O-acetyl-

4-phenoxymethyl-1,2,3-triazole (25s).

procedure C, 24 (40 mg, 0.10 mmol) was reacted with phenylprop-

argyl ether (19s, 26 L, 0.21 mmol), 0.2 M CuSO₄(103 L, 21 mol)

mol) to yield 25s (51 mg,

a

-D

-mannopyranosyl)methyl-

Following general

4.1.6.20. 1-(2,3,4,6-Tetra-O-acetyl-

4-(3-chlorophenyl)-1,2,3-triazole (25p).

procedure C, 24 (40 mg, 0.10 mmol) was reacted with 3-chloro-1-

a-

D-mannopyranosyl)methyl-

l

Following general

and 1 M sodium ascorbate (41

lL, 41 l

96%).

6468

O. Schwardt et al. / Bioorg. Med. Chem. 19 (2011) 6454–6473

[

a

]

+2.34 (c 1.03, CHCl₃); ¹H NMR (500 MHz, CDCl₃): d 2.00,

1H, H-2), 4.76, 4.87 (A, B of AB, J = 12.6 Hz, 1H, H-1⁰), 4.99 (d,

J = 0.7 Hz, 1H, H-1), 8.47 (t, J = 2.0 Hz, 1H, C₅H₃N-H2), 8.56 (m,

1H, C₂N₃H), 8.96 (d, J = 2.1 Hz, 1H, C₅H₃N-H6), 8.97 (d, J = 1.4 Hz,

D

2.08, 2.12, 2.13 (4 s, 12H, 4 COCH₃), 4.04 (dd, J = 4.1, 12.1 Hz, 1H,

H-6a), 4.15 (m, 1H, H-5), 4.37 (dt, J = 2.4, 8.6 Hz, 1H, H-1), 4.50

(dd, J = 8.7, 14.4 Hz, 1H, H-1⁰a), 4.55 (dd, J = 9.0, 12.1 Hz, 1H, H-

6b), 4.63 (dd, J = 2.5, 14.4 Hz, 1H, H-1⁰b), 4.96 (dd, J = 3.2, 4.9 Hz,

1H, H-4), 5.04 (dd, J = 3.3, 8.3 Hz, 1H, H-2), 5.34 (m, 1H, H-3),

6.96–6.99, 7.29–7.31 (m, 5H, C₆H₅), 7.80 (s, 1H, C₂N₃H); ¹³C NMR

(125 MHz, CDCl₃): d 20.49, 20.62, 20.66, 20.74 (4 COCH₃), 50.2

(C-1⁰), 60.0 (C-6), 61.9 (CH₂OPh), 66.7 (C-2), 66.9 (C-3), 67.8

(C-4), 68.5 (C-1), 73.3 (C-5), 114.6, 121.2 (3C, C₆H₅), 123.8

(C₂N₃H–C5), 129.5 (2C, C₆H₅), 144.3 (C₂N₃H–C4), 158.1 (C₆H₄–

C1), 169.1, 169.4, 169.6, 170.4 (4 COCH₃); ESI-MS Calcd for

13

1H, C₅H₃N-H4); C NMR (125 MHz, D₂O): d 59.7 (C-1⁰), 60.8 (C-

6), 66.7 (C-4), 69.9 (C-2), 70.4 (C-3), 73.0 (C-5), 99.5 (C-1), 123.6

(C₂N₃H–C5), 129.5, 133.1, 133.4, 142.6 (C₅H₃N), 144.6

(C₂N₃H–C4), 149.7 (C₅H₃N), 171.1 (CO); HR-MS Calcd for

C

₁₅H₁₇N₄Na₂O₈[M+Na]⁺: 427.0842, Found: 427.0844.

4.1.7.4. (1-Benzyl-1,2,3-triazol-4-yl)methyl

side (5d). Following general procedure A, 14 (40 mg,

0.18 mmol) was reacted with benzyl azide (15d, 34 L, 0.27 mmol),

CuSO₄(11 mg, 45 mol) and sodium ascorbate (18 mg, 90 mol) to

yield 5d (57 mg, 71%).

+53.3 (c 1.03, MeOH); ¹H NMR (500 MHz, CD₃OD): d 3.50

a-D-mannopyrano-

l

C

₂₄H₃₀N₃O₉[M+H]⁺: 520.19, Found: 520.16.

l

4.1.7. Synthesis of mannosyl triazoles

4.1.7.1. Sodium 4-[4-(( -mannopyranosyloxy)methyl)-1H-

1,2,3-triazol-1-yl]-benzoate (5a). To solution of 12a

(48.0 mg, 85.2 mol) in MeOH (4 mL) was added freshly prepared

1 M NaOMe in MeOH (0.4 mL) under argon. The mixture was stir-

red at rt for 3 h and then evaporated to dryness. The remains were

dissolved in H₂O/dioxane (1:1, 5 mL) and treated with 1 M aq.

NaOH (0.5 mL) for 16 h. The solution was concentrated and the res-

idue puriﬁed by MPLC on RP-18 (H₂O/MeOH) and P2 size exclusion

chromatography to give 5a (31.2 mg, 91%) as white powder after a

ﬁnal lyophilization from water.

[a]

D

a-

D

(m, 1H, H-5), 3.56 (t, J = 9.4 Hz, 1H, H-4), 3.60–3.68 (m, 2H, H-3,

H-6a), 3.73 (m, 1H, H-2), 3.79 (dd, J = 1.7, 11.7 Hz, 1H, H-6b),

4.60, 4.75 (A, B of AB, J = 12.4 Hz, 2H, H-1⁰), 4.80 (d, J = 1.6 Hz,

1H, H-1), 5.56 (s, 2H, CH₂Ph), 7.24–7.41 (m, 5H, C₆H₅), 7.97 (s,

1H, C₂N₃H); ¹³C NMR (125 MHz, CD₃OD): d 55.1 (CH₂Ph), 60.8 (C-

1⁰), 63.1 (C-6), 68.7 (C-4), 72.1 (C-2), 72.6 (C-3), 75.1 (C-5), 100.9

(C-1), 125.5 (C₂N₃H–C5), 129.3, 129.8, 130.2, 136.9 (6C, C₆H₅),

145.9 (C₂N₃H–C4); HR-MS Calcd for C₁₆H₂₁N₃NaO₆[M+Na]⁺:

374.1328, Found: 374.1334.

a

l

[

a

]

+41.7 (c 0.60, H₂O); IR (KBr) 3413 (vs b, OH), 1607 (vs,

4.1.7.5. [1-(4⁰-Aminophenyl)-1,2,3-triazol-4-yl]methyl

nopyranoside hydrochloride (5e). Following general proce-

dure A, 14 (40 mg, 0.18 mmol) was reacted with 4-azidoaniline

hydrochloride (15e, 46 mg, 0.27 mmol), CuSO₄(11 mg, 45 mol)

and sodium ascorbate (18 mg, 90 mol) to yield 5e (19 mg, 27%).

_D+55.2 (c 1.00, MeOH); ¹H NMR (500 MHz, CD₃OD): d 3.55–

3.60 (m, 2H, H-4, H-5), 3.64–3.72 (m, 2H, H-3, H-6a), 3.78 (m, 1H,

H-2), 3.83 (m, 1H, H-6b), 4.68, 4.82 (A, B of AB, J = 12.4 Hz, 2H, H-

1⁰), 4.86 (m, 1H, H-1), 6.78, 7.45 (AA⁰, BB⁰of AA⁰BB⁰, J = 8.7 Hz, 4H,

C₆H₄), 8.33 (s, 1H, C₂N₃H); ¹³C NMR (125 MHz, CD₃OD): d 60.8

(C-1⁰), 63.2 (C-6), 68.8 (C-4), 72.2 (C-2), 72.6 (C-3), 75.2 (C-5),

100.9 (C-1), 116.2 (2C, C₆H₄), 123.3 (2C, C₆H₄), 123.7 (C₂N₃H–C5),

128.8 (C₆H₄–C1), 145.8 (C₂N₃H–C4), 150.8 (C₆H₄–C4); HR-MS Calcd

for C₁₅H₂₁N₄O₆[M+H]⁺: 353.1461, Found: 353.1463.

a-D-man-

D

CO) cm^ꢁ^{1 1}H NMR (500 MHz, D₂O): d 3.60–3.66 (m, 2H, H-4, H-

;

5), 3.72 (dd, J = 5.5, 12.1 Hz, 1H, H-6a), 3.78 (dd, J = 3.4, 9.4 Hz,

1H, H-3), 3.82 (dd, J = 1.4, 12.2 Hz, 1H, H-6b), 3.93 (dd, J = 1.7,

3.4 Hz, 1H, H-2), 4.71 (m, 1H, H-1⁰a), 4.82 (m, 1H, H-1⁰b), 4.97 (s,

1H, H-1), 7.68 (m, 2H, C₆H₄), 7.94 (m, 2H, C₆H₄), 8.41 (m, 1H,

l

[a]

13

C₂N₃H); C NMR (125 MHz, D₂O): d 59.7 (C-1⁰), 60.8 (C-6), 66.6

(C-4), 69.9 (C-2), 70.4 (C-3), 72.9 (C-5), 99.5 (C-1), 120.2 (2C,

C₆H₄), 123.3 (C₂N₃H–C5), 130.3, 137.0, 137.7 (4C, C₆H₄), 144.2

(C₂N₃H–C4), 174.1 (CO); HR-MS Calcd for C₁₆H₁₉N₃NaO₈[M+H]⁺:

404.1070, Found: 404.1071.

4.1.7.2. Sodium 3-[4-((

1,2,3-triazol-1-yl]-benzoate (5b).

a-

D

-mannopyranosyloxy)methyl)-1H-

According to the proce-

dure described for 5a, 12b (67.0 mg, 0.116 mmol) was subse-

quently treated with 1 M methanolic NaOMe (0.5 mL) in MeOH

(5 mL) and 1 M aq. NaOH (0.5 mL) in H₂O/dioxane (1:1, 6 mL) to

yield 5b (37.7 mg, 81%).

4.1.7.6. (1-Adamantyl-1,2,3-triazol-4-yl)methyl

ranoside (5f). Following general procedure A, 14 (40 mg,

0.18 mmol) was reacted with 1-azidoadamantane (15f, 48 mg,

0.27 mmol), CuSO₄(11 mg, 45 mol) and sodium ascorbate

(18 mg, 90 mol) to yield 5f (20 mg, 28%).

_D+50.5 (c 1.04, MeOH); ¹H NMR (500 MHz, CD₃OD): d 1.76–

a-D-mannopy-

[

a

]

D

+44.4 (c 0.89, H₂O); IR (KBr) 3401 (vs b, OH), 1610 (vs,

;

l

CO) cm^ꢁ1

¹H NMR (500 MHz, D₂O): d 3.61–3.66 (m, 2H, H-4,

l

H-5), 3.72 (dd, J = 5.4, 12.2 Hz, 1H, H-6a), 3.78 (dd, J = 3.5, 9.5 Hz,

1H, H-3), 3.82 (dd, J = 1.5, 12.2 Hz, 1H, H-6b), 3.92 (dd, J = 1.7,

3.3 Hz, 1H, H-2), 4.69 (A of AB, J = 12.8 Hz, 1H, H-1⁰a), 4.80 (m,

1H, H-1⁰b), 4.96 (s, 1H, H-1), 7.51 (m, 1H, C₆H₄–H5), 7.69 (d,

J = 7.0 Hz, 1H, C₆H₄–H4), 7.87 (d, J = 7.7 Hz, 1H, C₆H₄-H6), 8.01 (s,

1H, C₆H₄-H2), 8.36 (m, 1H, C₂N₃H); ¹³C NMR (125 MHz, D₂O): d

59.7 (C-1⁰), 60.8 (C-6), 66.7 (C-4), 69.9 (C-2), 70.4 (C-3), 72.9 (C-

5), 99.5 (C-1), 120.8 (C₆H₄), 123.0 (C₂N₃H–C5), 123.3, 129.5,

129.8, 135.9, 138.1 (C₆H₄), 144.1 (C₂N₃H–C4), 173.6 (CO); HR-MS

Calcd for C₁₆H₁₉N₃NaO₈[M+H]⁺: 404.1070, Found: 404.1068.

[a]

1.88 (m, 6H, Ad), 2.24 (s, 9H, Ad), 3.50–3.60 (m, 2H, H-4, H-5),

3.61–3.70 (m, 2H, H-3, H-6a), 3.75 (dd, J = 1.7, 3.3 Hz, 1H, H-2),

3.82 (m, 1H, H-6b), 4.60, 4.76 (A, B of AB, J = 12.3 Hz, 2H, H-1⁰),

4.80 (d, J = 1.4 Hz, 1H, H-1), 8.09 (s, 1H, C₂N₃H); ¹³C NMR

(125 MHz, CD₃OD): d 31.1, 37.1, 44.0 (10 C, Ad), 60.9 (C-1⁰), 63.2

(C-6), 68.8 (C-4), 72.2 (C-2), 72.6 (C-3), 75.1 (C-5), 100.9 (C-1),

122.2 (C₂N₃H–C5), 144.6 (C₂N₃H–C4); HR-MS Calcd for

C

₁₉H₂₉N₃NaO₆[M+Na]⁺: 418.1954, Found: 418.1951.

4.1.7.7. [1-(4⁰-Methoxybenzyl)-1,2,3-triazol-4-yl]methyl

mannopyranoside (5g). Following general procedure A, 14

(50 mg, 0.23 mmol) was reacted with 4-methoxybenzylazide

(15g,³⁸57 mg, 0.35 mmol), CuSO₄(15 mg, 60

mol) and sodium

ascorbate (24 mg, 120 mol) to yield 5g (64 mg, 73%).

+66.6 (c 1.01, MeOH); ¹H NMR (500 MHz, CD₃OD): d 3.50

(m, 1H, H-5), 3.57 (t, J = 9.4 Hz, 1H, H-4), 3.61–3.69 (m, 2H, H-3,

H-6a), 3.74 (m, 4H, H-2, OCH₃), 3.78 (dd, J = 1.7, 11.7 Hz, 1H, H-

6b), 4.58, 4.73 (A, B of AB, J = 12.4 Hz, 2H, H-1⁰), 4.79 (m, 1H, H-

1), 5.46 (s, 2H, CH₂Ar), 6.88, 7.25 (AA⁰, BB⁰of AA⁰BB⁰, J = 8.6 Hz,

4H, C₆H₄), 7.91 (s, 1H, C₂N₃H);

a-D-

4.1.7.3. Sodium 5-[4-((a-D-mannopyranosyloxy)methyl)-1H-

1,2,3-triazol-1-yl]-nicotinate (5c).

According to the proce-

dure described for 5a, 12c (41.0 mg, 72.6

lmol) was subsequently

l

treated with 1 M methanolic NaOMe (0.4 mL) in MeOH (4 mL) and

1 M aq. NaOH (0.4 mL) in H₂O/dioxane (1:1, 4 mL) to yield 5c

(23.0 mg, 78%).

l

[a]

D

[a] +36.0 (c 0.69, H₂O); IR (KBr) 3413 (vs b, OH), 1616 (vs,

D

CO) cm^ꢁ^{1 1}H NMR (500 MHz, D₂O): d 3.61–3.66 (m, 2H, H-4, H-

;

5), 3.73 (dd, J = 5.4, 12.1 Hz, 1H, H-6a), 3.79 (dd, J = 3.5, 9.4 Hz,

1H, H-3), 3.83 (d, J = 12.0 Hz, 1H, H-6b), 3.94 (dd, J = 1.7, 3.2 Hz,

O. Schwardt et al. / Bioorg. Med. Chem. 19 (2011) 6454–6473

6469

¹³C NMR (125 MHz, CD₃OD): d 54.6 (CH₂Ar), 55.8 (OCH₃), 60.7

4.1.7.12.

mannopyranoside (5l).

[1-(3⁰-Fluorophenyl)-1,2,3-triazol-4-yl]methyl

a-D-

(C-1⁰), 63.0 (C-6), 68.6 (C-4), 72.0 (C-2), 72.5 (C-3), 75.0 (C-5),

100.8 (C-1), 115.4 (2C, C₆H₄), 125.1 (C₂N₃H–C5), 128.6 (2C, C₆H₄),

130.8 (C₆H₄–C1), 145.6 (C₂N₃H–C4), 161.4 (C₆H₄–C4); HR-MS Calcd

for C₁₇H₂₃N₃NaO₇[M+Na]⁺: 404.1434, Found: 404.1431.

Prepared from 16l (105 mg, 0.20

mmol) according to general procedure D. Yield: 58 mg, 81%.

[a]

D

+73.8 (c 1.00, MeOH); ¹H NMR (500 MHz, CD₃OD): d 3.51–

3.62 (m, 2H, H-4, H-5), 3.62–3.73 (m, 2H, H-3, H-6a), 3.78 (dd,

J = 1.6, 3.1 Hz, 1H, H-2), 3.84 (m, 1H, H-6b), 4.71 (A of AB,

J = 12.4 Hz, 1H, H-1⁰a), 4.82–4.89 (m, 2H, H-1, H-1⁰b), 7.23 (m,

1H, C₆H₄), 7.57 (m, 1H, C₆H₄), 7.64–7.74 (m, 2H, C₆H₄), 8.61 (s,

4.1.7.8. [1-(3⁰-Methoxybenzyl)-1,2,3-triazol-4-yl]methyl

mannopyranoside (5h). Following general procedure A, 14

(50 mg, 0.23 mmol) was reacted with 3-methoxybenzylazide

(15h,³⁹57 mg, 0.35 mmol), CuSO₄(15 mg, 60

mol) and sodium

ascorbate (24 mg, 120 mol) to yield 5h (68 mg, 77%).

+62.2 (c 1.00, MeOH); ¹H NMR (500 MHz, CD₃OD): d 3.52

a-D-

13

1H, C₂N₃H); C NMR (125 MHz, CD₃OD): d 60.8 (C-1⁰), 63.1 (C-

l

6), 68.8 (C-4), 72.1 (C-2), 72.6 (C-3), 75.2 (C-5), 101.1 (C-1), 109.1

(d, J = 26.3 Hz, C₆H₄), 116.9 (d, J = 21.3 Hz, C₆H₄), 117.3 (d,

J = 3.8 Hz, C₆H₄), 123.8 (C₂N₃H–C5), 132.9 (d, J = 10.0 Hz, C₆H₄),

139.7 (d, J = 10.0 Hz, C₆H₄–C1), 146.6 (C₂N₃H–C4), 161.7 (d,

J = 245.0 Hz, C₆H₄–C3); HR-MS Calcd for C₁₅H₁₈FN₃NaO₆[M+Na]⁺:

378.1077, Found: 378.1081.

l

[a]

D

(m, 1H, H-5), 3.58 (t, J = 9.4 Hz, 1H, H-4), 3.61–3.70 (m, 2H, H-3,

H-6a), 3.70–3.84 (m, 5H, H-2, H-6b, OCH₃), 4.60, 4.75 (A, B of AB,

J = 12.3 Hz, 2H, H-1⁰), 4.81 (m, 1H, H-1), 5.52 (s, 2H, CH₂Ar), 6.85

(s, 3H, C₆H₄), 7.24 (t, J = 7.9 Hz, 1H, C₆H₄), 7.98 (s, 1H, C₂N₃H);

¹³C NMR (125 MHz, CD₃OD): d 55.5 (CH₂Ar), 55.9 (OCH₃), 60.8

(C-1⁰), 63.0 (C-6), 68.7 (C-4), 72.1 (C-2), 72.6 (C-3), 75.1 (C-5),

100.9 (C-1), 114.9, 115.2, 121.4 (3C, C₆H₄), 125.6 (C₂N₃H–C5),

131.3 (C₆H₄), 138.2 (C₆H₄–C1), 145.8 (C₂N₃H–C4), 161.7 (C₆H₄–

C3); HR-MS Calcd for C₁₇H₂₃N₃NaO₇[M+Na]⁺: 404.1434, Found:

404.1435.

4.1.7.13. [1-(4⁰-Methoxyphenyl)-1,2,3-triazol-4-yl]methyl

mannopyranoside (5m). Prepared from 16m (113 mg,

0.21 mmol) according to general procedure D. Yield: 58 mg, 75%.

+37.4 (c 1.01, MeOH); ¹H NMR (500 MHz, CD₃OD): d

a-D-

[a]

D

3.54–3.64 (m, 2H, H-4, H-5), 3.65–3.74 (m, 2H, H-3, H-6a),

3.80 (dd, J = 1.6, 3.2 Hz, 1H, H-2), 3.81–3.87 (m, 4H, H-6b,

OCH₃), 4.69, 4.83 (A, B of AB, J = 12.4 Hz, 2H, H-1⁰), 4.87 (m,

1H, H-1), 7.05, 7.68 (AA⁰, BB⁰of AA⁰BB⁰, J = 9.0 Hz, 4H, C₆H₄),

4.1.7.9. [1-(4⁰-Nitrophenyl)-1,2,3-triazol-4-yl]methyl

nopyranoside (5i). Following general procedure A, 14 (40 mg,

0.18 mmol) was reacted with 1-azido-4-nitrobenzene (15i,⁴⁰

44 mg, 0.27 mmol), CuSO₄(11 mg, 45 mol) and sodium ascorbate

(18 mg, 90 mol) to yield 5i (31 mg, 44%).

+50.4 (c 1.02, MeOH); ¹H NMR (500 MHz, CD₃OD): d 3.56–

a-D-man-

8.42 (s, 1H, C₂N₃H); ¹³C NMR (125 MHz, CD₃OD):

d 56.3

(OCH₃), 60.8 (C-1⁰), 63.1 (C-6), 68.8 (C-4), 72.2 (C-2), 72.6 (C-

3), 75.2 (C-5), 101.0 (C-1), 116.0 (2C, C₆H₄), 123.4 (2C, C₆H₄),

123.8 (C₂N₃H–C5), 131.7 (C₆H₄–C1), 146.1 (C₂N₃H–C4), 161.7

(C₆H₄–C4); HR-MS Calcd for C₁₆H₂₁N₃NaO₇[M+Na]⁺: 390.1277,

Found: 390.1279.

l

[a]

D

3.60 (m, 2H, H-4, H-5), 3.64–3.72 (m, 2H, H-3, H-6a), 3.80 (dd,

J = 1.7, 3.3 Hz, 1H, H-2), 3.84 (dd, J = 1.0, 11.7 Hz, 1H, H-66), 4.75

(A of AB, J = 12.5 Hz, 1H, H-1⁰a), 4.88–4.91 (m, 2H, H-1, H-1⁰b),

8.16 (m, 2H, C₆H₄), 8.44 (m, 2H, C₆H₄), 8.75 (s, 1H, C₂N₃H); ¹³C

NMR (125 MHz, CD₃OD): d 60.9 (C-1⁰), 63.2 (C-6), 68.8 (C-4), 72.1

(C-2), 72.6 (C-3), 75.3 (C-5), 101.2 (C-1), 122.0 (2C, C₆H₄), 123.9

(C₂N₃H–C5), 126.7 (2C, C₆H₄), 142.7 (C₆H₄–C1), 147.1 (C₂N₃H–

C4), 148.9 (C₆H₄–C4); HR-MS Calcd for C₁₅H₁₈N₄NaO₈[M+Na]⁺:

405.1022, Found: 405.1020.

4.1.7.14. [1-(3⁰-Methoxybenzyl)-1,2,3-triazol-4-yl]ethyl

nopyranoside (6h). Prepared from 17h (53 mg, 94

according to general procedure D. Yield: 30 mg, 81%.

_D+45.9 (c 1.00, MeOH); ¹H NMR (500 MHz, CD₃OD): d 2.99 (t,

a-

D

-man

mol)

l

[a]

J = 6.6 Hz, 2H, H-2⁰), 3.41 (m, 1H, H-5), 3.57–3.75 (m, 4H, H-3, H-4,

H-6a, H-1⁰a), 3.71 (dd, J = 1.7, 3.1 Hz, 1H, H-2), 3.78–3.83 (m, 4H, H-

6b, OCH₃), 3.97 (dt, J = 6.7, 9.7 Hz, 1H, H-1⁰b), 4.77 (d, J = 1.6 Hz, 1H,

H-1), 5.54 (s, 2H, CH₂Ar), 6.80–6.96, 7.29 (m, 4H, C₆H₄), 7.79 (s, 1H,

4.1.7.10. [1-(Pyridin-4⁰-yl)-1,2,3-triazol-4-yl]methyl

pyranoside (5j). Prepared from 16j (102 mg, 0.20 mmol)

according to general procedure D. Yield: 58 mg, 85%.

+70.3 (c 1.00, MeOH); ¹H NMR (500 MHz, CD₃OD): d 3.53–

a-D-manno-

13

C₂N₃H); C NMR (125 MHz, CD₃OD): d 27.2 (C-2⁰), 54.9 (CH₂Ar),

55.9 (OCH₃), 63.0 (C-6), 67.6 (C-1⁰), 68.7 (C-4), 72.2 (C-2), 72.7

(C-3), 74.9 (C-5), 101.7 (C-1), 114.8, 115.1, 121.2 (3C, C₆H₄),

124.2 (C₂N₃H–C5), 131.3 (C₆H₄), 138.4 (C₆H₄–C1), 146.7 (C₂N₃H–

C4), 161.7 (C₆H₄–C3); HR-MS Calcd for C₁₈H₂₅N₃NaO₇[M+Na]⁺:

418.1590, Found: 418.1591.

[a]

D

3.63 (m, 2H, H-4, H-5), 3.64–3.74 (m, 2H, H-3, H-6a), 3.77–3.87 (m,

2H, H-2, H-6b), 4.73 (A of AB, J = 12.6 Hz, 1H, H-1⁰a), 4.84–4.92 (m,

2H, H-1, H-1⁰b), 7.96, 8.67 (m, 4H, C₅H₄N), 8.77 (s, 1H, C₂N₃H); ¹³C

NMR (125 MHz, CD₃OD): d 60.9 (C-1⁰), 63.1 (C-6), 68.7 (C-4), 72.1

(C-2), 72.6 (C-3), 75.2 (C-5), 101.2 (C-1), 115.5 (2C, C₅H₄N), 123.5

(C₂N₃H–C5), 145.2 (C₂N₃H–C4), 147.2 (C₅H₄N–C1), 152.5 (2C,

C₅H₄N); HR-MS Calcd for C₁₄H₁₉N₄O₆[M+H]⁺: 339.1305, Found:

339.1302.

4.1.7.15. [1-(4⁰-Nitrophenyl)-1,2,3-triazol-4-yl]ethyl

pyranoside (6i). Prepared from 17i (61 mg, 0.11 mmol)

according to general procedure D. Yield: 37 mg, 86%.

_D+44.4 (c 1.00, MeOH); ¹H NMR (500 MHz, CD₃OD): d 3.13 (t,

a-D-manno-

[a]

J = 6.5 Hz, 2H, H-2⁰), 3.42 (m, 1H, H-5), 3.60 (t, J = 9.5 Hz, 1H, H-4),

3.64–3.73 (m, 2H, H-3, H-6a), 3.78–3.88 (m, 3H, H-2, H-6b, H-1⁰a),

4.09 (dt, J = 6.6, 9.8 Hz, 1H, H-1⁰b), 4.83 (d, J = 1.5 Hz, 1H, H-1), 8.18,

8.48 (m, 4H, C₆H₄), 8.55 (s, 1H, C₂N₃H); ¹³C NMR (125 MHz,

CD₃OD): d 27.3 (C-2⁰), 63.1 (C-6), 67.4 (C-1⁰), 68.7 (C-4), 72.2 (C-

2), 72.8 (C-3), 75.0 (C-5), 101.8 (C-1), 122.0 (2C, C₆H₄), 122.5

(C₂N₃H–C5), 126.6 (2C, C₆H₄), 142.8 (C₆H₄–C1), 148.1 (C₂N₃H–

C4), 148.9 (C₆H₄–C4); HR-MS Calcd for C₁₆H₂₀N₄NaO₈[M+Na]⁺:

419.1179, Found: 419.1177.

4.1.7.11. [1-(4⁰-Fluorophenyl)-1,2,3-triazol-4-yl]methyl

nopyranoside (5k). Prepared from 16k (106 mg, 0.20 mmol)

according to general procedure D. Yield: 56 mg, 78%.

+78.5 (c 1.00, MeOH); ¹H NMR (500 MHz, CD₃OD): d 3.60–

a-D-man

[a]

D

3.69 (m, 2H, H-4, H-5), 3.71–3.78 (m, 2H, H-3, H-6a), 3.86 (dd,

J = 1.7, 3.4 Hz, 1H, H-2), 3.89 (dd, J = 1.8, 11.8 Hz, 1H, H-6b), 4.77,

4.91 (A, B of AB, J = 12.5 Hz, 2H, H-1⁰), 4.94 (d, J = 1.6 Hz, 1H, H-

1), 7.34, 7.89 (m, 4H, C₆H₄), 8.56 (s, 1H, C₂N₃H); ¹³C NMR

(125 MHz, CD₃OD): d 60.9 (C-1⁰), 63.2 (C-6), 68.8 (C-4), 72.2 (C-

2), 72.6 (C-3), 75.2 (C-5), 101.1 (C-1), 117.8 (d, J = 23.8 Hz, 2C,

C₆H₄), 124.0 (2C, C₆H₄), 124.0 (C₂N₃H–C5), 134.9 (d, J = 3.8 Hz,

C₆H₄–C1), 146.5 (C₂N₃H–C4), 164.1 (d, J = 246.3 Hz, C₆H₄–C4);

4.1.7.16. [1-(Pyridin-4⁰-yl)-1,2,3-triazol-4-yl]ethyl

pyranoside (6j). Prepared from 17j (63 mg, 0.12 mmol)

according to general procedure D. Yield: 31 mg, 73%.

_D+48.3 (c 1.00, MeOH); ¹H NMR (500 MHz, CD₃OD): d 3.12 (t,

J = 6.5 Hz, 2H, H-2⁰), 3.43 (m, 1H, H-5), 3.61 (t, J = 9.5 Hz, 1H, H-4),

a-D-manno-

HR-MS Calcd for

378.1079.

C

₁₅H₁₈FN₃NaO₆[M+Na]⁺: 378.1077, Found:

[a]

6470

O. Schwardt et al. / Bioorg. Med. Chem. 19 (2011) 6454–6473

3.64–3.73 (m, 2H, H-3, H-6a), 3.77–3.87 (m, 3H, H-2, H-6b, H-1⁰a),

4.08 (dt, J = 6.6, 9.8 Hz, 1H, H-1⁰b), 4.83 (d, J = 1.5 Hz, 1H, H-1), 7.99

(dd, J = 1.6, 4.8 Hz, 2H, C₅H₄N), 8.58 (s, 1H, C₂N₃H), 8.74 (m, 2H,

4.1.7.21. 4-(4-Triﬂuoromethylphenyl)-1-(

1,2,3-triazole (7q). Prepared from 20q (46 mg, 85

according to general procedure D. Yield: 27 mg, 86%.

+83.4 (c 0.34, MeOH); ¹H NMR (500 MHz, CD₃OD): d 3.38

a

-

D

-mannopyranosyl)-

lmol)

13

C₅H₄N); C NMR (125 MHz, CD₃OD): d 27.2 (C-2⁰), 63.1 (C-6),

[a]

D

67.3 (C-1⁰), 68.7 (C-4), 72.2 (C-2), 72.7 (C-3), 75.0 (C-5), 101.8 (C-

1), 115.5 (2C, C₅H₄N), 122.0 (C₂N₃H–C5), 145.3 (C₅H₄N–C1),

148.1 (C₂N₃H–C4), 152.4 (2C, C₅H₄N); HR-MS Calcd for

(m, 1H, H-5), 3.77–3.81 (m, 2H, H-4, H-6a), 3.85 (dd, J = 1.9,

12.0 Hz, 1H, H-6b), 4.11 (dd, J = 3.3, 8.4 Hz, 1H, H-3), 4.76 (t,

J = 2.7 Hz, 1H, H-2), 6.10 (d, J = 2.3 Hz, 1H, H-1), 7.76, 8.06 (AA⁰,

BB⁰of AA⁰BB⁰, J = 8.0 Hz, 4H, C₆H₄), 8.66 (s, 1H, C₂N₃H); ¹³C NMR

(125 MHz, CD₃OD): d 62.6 (C-6), 68.7 (C-4), 70.1 (C-2), 72.6 (C-3),

78.8 (C-5), 88.6 (C-1), 123.2 (C₂N₃H–C5), 125.6 (q, J = 272 Hz,

CF₃), 127.0 (q, J = 3.7 Hz, 2C, C₆H₄–C3, C5), 127.2 (2C, C₆H₄–C2,

C6), 131.2 (d, J = 32.4 Hz, C₆H₄–C4), 135.5 (C₆H₄–C1), 147.6

(C₂N₃H–C4); HR-MS Calcd for C₁₅H₁₆F₃N₃NaO₅[M+Na]⁺:

398.0940, Found: 398.0942.

C

₁₅H₂₁N₄O₆[M+H]⁺: 353.1461, Found: 353.1460.

4.1.7.17. [1-(4⁰-Fluorophenyl)-1,2,3-triazol-4-yl]ethyl

nopyranoside (6k). Prepared from 17k (65 mg, 0.12 mmol)

according to general procedure D. Yield: 40 mg, 90%.

_D+50.7 (c 1.00, MeOH); ¹H NMR (500 MHz, CD₃OD): d 3.10 (t,

a-D-man-

[a]

J = 6.5 Hz, 2H, H-2⁰), 3.43 (m, 1H, H-5), 3.61 (t, J = 9.5 Hz, 1H, H-4),

3.65–3.74 (m, 2H, H-3, H-6a), 3.77–3.87 (m, 3H, H-2, H-6b, H-1⁰a),

4.07 (dt, J = 6.6, 9.8 Hz, 1H, H-1⁰b), 4.82 (d, J = 1.6 Hz, 1H, H-1), 7.34,

7.88 (m, 4H, C₆H₄), 8.33 (s, 1H, C₂N₃H); ¹³C NMR (125 MHz,

CD₃OD): d 27.2 (C-2⁰), 63.1 (C-6), 67.5 (C-1⁰), 68.7 (C-4), 72.2 (C-

2), 72.8 (C-3), 75.0 (C-5), 101.8 (C-1), 117.8 (d, J = 23.8 Hz, 2C,

C₆H₄), 122.6 (C₂N₃H–C5), 124.0 (d, J = 8.8 Hz, 2C, C₆H₄), 135.0 (d,

J = 2.5 Hz, C₆H₄–C1), 147.4 (C₂N₃H–C4), 164.1 (d, J = 246.3 Hz,

C₆H₄–C4); HR-MS Calcd for C₁₆H₂₀FN₃NaO₆[M+Na]⁺: 392.1234,

Found: 392.1238.

4.1.7.22.

(7r).

1-(

a

-

D

-Mannopyranosyl)-4-(3-pyridyl)-1,2,3-triazole

mol) according to gen-

Prepared from 20r (47 mg, 98

l

eral procedure D. Yield: 28 mg, 92%.

[a]

D

+86.7 (c 0.93, MeOH); ¹H NMR (500 MHz, CD₃OD): d 3.40

(ddd, J = 2.3, 6.6, 8.6 Hz, 1H, H-5), 3.77–3.82 (m, 2H, H-4, H-6a),

3.86 (dd, J = 2.4, 12.1 Hz, 1H, H-6b), 4.11 (dd, J = 3.5, 8.4 Hz, 1H,

H-3), 4.76 (t, J = 3.1 Hz, 1H, H-2), 6.11 (d, J = 2.7 Hz, 1H, H-1),

7.54 (dd, J = 5.0, 7.8 Hz, 1H, C₅H₄N-H5), 8.31 (m, 1H, C₅H₄N-H6),

8.53 (dd, J = 1.4, 4.9 Hz, 1H, C₅H₄N-H4), 8.68 (s, 1H, C₂N₃H), 9.04

(d, J = 1.5, 1H, C₅H₄N–H2); ¹³C NMR (125 MHz, CD₃OD): d 62.6

(C-6), 68.7 (C-4), 70.1 (C-2), 72.6 (C-3), 78.8 (C-5), 88.6 (C-1),

123.1 (C₂N₃H–C5), 125.6 (C₅H₄N–C5), 128.5 (C₅H₄N–C1), 135.0

(C₅H₄N–C6), 145.6 (C₂N₃H–C4), 147.3 (C₅H₄N–C2), 149.7 (C₅H₄N–

C4); HR-MS Calcd for C₁₃H₁₆N₄NaO₅[M+Na]⁺: 331.1018, Found:

331.1013.

4.1.7.18.

(7n).

1-(

a-D-Mannopyranosyl)-4-phenyl-1,2,3-triazole

Prepared from 20n (50 mg, 0.11 mmol) according to

general procedure D. Yield: 29 mg, 89%.

[a]

D

+98.0 (c 1.34, MeOH); ¹H NMR (500 MHz, CD₃OD): d 3.38

(ddd, J = 2.5, 6.6, 8.9 Hz, 1H, H-5), 3.76–3.80 (m, 2H, H-4, H-6a),

3.85 (dd, J = 2.5, 12.1 Hz, 1H, H-6b), 4.12 (dd, J = 3.5, 8.5 Hz, 1H, H-

3), 4.76 (t, J = 3.1 Hz, 1H, H-2), 6.08 (d, J = 2.7 Hz, 1H, H-1), 7.34–

7.38, 7.43–7.46, 7.84–7.85 (m, 5H, C₆H₅), 8.51 (s, 1H, C₂N₃H); ¹³C

NMR (125 MHz, CD₃OD): d 62.6 (C-6), 68.7 (C-4), 70.1 (C-2), 72.6

(C-3), 78.7 (C-5), 88.5 (C-1), 122.1 (C₂N₃H–C5), 126.8, 129.5, 130.0,

131.4 (6C, C₆H₅), 149.0 (C₂N₃H–C4); HR-MS Calcd for C₁₄H₁₇N₃NaO₅

[M+Na]⁺: 330.1066, Found: 330.1060.

4.1.7.23. 1-(

zole (7s).

a

-

D

-Mannopyranosyl)-4-phenoxymethyl-1,2,3-tria-

Prepared from 20s (46 mg, 90 mol) according to

l

general procedure D. Yield: 27 mg, 89%.

[a]

D

+57.0 (c 0.90, MeOH); ¹H NMR (500 MHz, CD₃OD): d 3.30

(m, 1H, H-5), 3.70–3.74 (m, 2H, H-4, H-6a), 3.78 (dd, J = 1.8,

12.1 Hz, 1H, H-6b), 4.04 (dd, J = 3.2, 8.4 Hz, 1H, H-3), 4.67 (m, 1H,

H-2), 5.14 (s, 2H, CH₂OPh), 6.00 (d, J = 1.7 Hz, 1H, H-1), 6.91 (t,

J = 7.3 Hz, 1H, C₆H₅-H4), 6.96 (d, J = 8.1 Hz, 2H, C₆H₅-H2, H6),

7.24 (t, J = 7.8 Hz, 2H, C₆H₅-H3, H5), 8.20 (s, 1H, C₂N₃H); ¹³C NMR

(125 MHz, CD₃OD): d 62.2 (CH₂OPh), 62.5 (C-6), 68.6 (C-4), 70.1

(C-2), 72.5 (C-3), 78.6 (C-5), 88.4 (C-1), 115.8 (2C, C₆H₅–C2, C6),

122.3 (C₆H₅–C4), 125.3 (C₂N₃H–C5), 130.5 (2C, C₆H₅–C3, C5),

145.4 (C₂N₃H–C4), 159.7 (C₆H₅–C1); HR-MS Calcd for

4.1.7.19. 1-(

azole (7o).

a-

D

-Mannopyranosyl)-4-(4-methylphenyl)-1,2,3-tri-

Prepared from 20o (46 mg, 94 mol) according to

l

general procedure D. Yield: 20 mg, 65%.

[a]

_D+84.6 (c 0.63, MeOH); ¹H NMR (500 MHz, CD₃OD): d 2.36 (s,

3H, PhCH₃), 3.36 (ddd, J = 2.3, 6.7, 8.9 Hz, 1H, H-5), 3.74–3.78 (m,

2H, H-4, H-6a), 3.84 (dd, J = 2.4, 12.1 Hz, 1H, H-6b), 4.10 (dd,

J = 3.5, 8.5 Hz, 1H, H-3), 4.74 (t, J = 3.0 Hz, 1H, H-2), 6.05 (d,

J = 2.6 Hz, 1H, H-1), 7.25, 7.72 (AA⁰, BB⁰of AA⁰BB⁰, J = 8.0 Hz, 4H,

C₆H₄), 8.45 (s, 1H, C₂N₃H); ¹³C NMR (125 MHz, CD₃OD): d 21.3

(PhCH₃), 62.6 (C-6), 68.7 (C-4), 70.1 (C-2), 72.6 (C-3), 78.7 (C-5),

88.5 (C-1), 121.7 (C₂N₃H–C5), 126.7, 128.6, 130.6, 139.6 (6C,

C₆H₅), 149.2 (C₂N₃H–C4); HR-MS Calcd for C₁₅H₁₈N₃NaO₅

[M+Na]⁺: 344.1222, Found: 344.1215.

C

₁₅H₁₉N₃NaO₅[M+Na]⁺: 360.1172, Found: 360.1171.

4.1.7.24.

1-(

a-D

-Mannopyranosyl)methyl-4-phenyl-1,2,3-tria-

zole (8n).

Prepared from 25n (38 mg, 78 mol) according to

l

general procedure D. Yield: 22 mg, 87%.

[a]

_D+30.6 (c 0.91, MeOH); ¹H NMR (500 MHz, CD₃OD): d 3.73–

3.75 (m, 2H, H-4, H-6a), 3.79–3.85 (m, 3H, H-2, H-3, H-5), 3.88 (dd,

J = 7.2, 11.5 Hz, 1H, H-6b), 4.25 (dt, J = 4.8, 7.9 Hz, 1H, H-1), 4.73

(dd, J = 8.0, 14.4 Hz, 1H, H-1⁰a), 4.76 (dd, J = 4.5, 14.3 Hz, 1H, H-

1⁰b), 7.33 (t, J = 7.7 Hz, 1H, C₆H₅-H4), 7.42 (t, J = 7.8 Hz, 2H, C₆H₅-

H3, H5), 7.81 (d, J = 8.0 Hz, 2H, C₆H₅-H2, H6), 8.45 (s, 1H, C₂N₃H);

¹³C NMR (125 MHz, CD₃OD): d 50.9 (C-1⁰), 62.1 (C-6), 69.1 (C-2),

70.0 (C-4), 72.5 (C-3), 74.9 (C-1), 78.5 (C-5), 123.4 (C₂N₃H–C5),

126.7, 129.3, 129.9, 131.8 (6C, C₆H₅), 148.8 (C₂N₃H–C4); HR-MS

Calcd for C₁₅H₁₉NaN₃O₅[M+Na]⁺: 344.1222, Found: 344.1222.

4.1.7.20. 4-(3-Chlorophenyl)-1-(

azole (7p). Prepared from 20p (43 mg, 84

general procedure D. Yield: 25 mg, 87%.

+89.2 (c 0.50, MeOH); ¹H NMR (500 MHz, CD₃OD): d 3.37

a-

D-mannopyranosyl)-1,2,3-tri-

l

mol) according to

[a]

D

(ddd, J = 2.4, 6.7, 8.7 Hz, 1H, H-5), 3.77 (dd, J = 6.6, 12.2 Hz, 1H,

H-6a), 3.78 (t, J = 8.6 Hz, 1H, H-4), 3.87 (dd, J = 2.4, 12.1 Hz, 1H,

H-6b), 4.11 (dd, J = 3.5, 8.3 Hz, 1H, H-3), 4.75 (t, J = 3.1 Hz, 1H,

H-2), 6.07 (d, J = 2.6 Hz, 1H, H-1), 7.37 (d, J = 8.1 Hz, 1H, C₆H₄-

H6), 7.44 (t, J = 7.9 Hz, 1H, C₆H₄–H5), 7.79 (d, J = 7.7 Hz, 1H,

C₆H₄–H4), 7.90 (s, 1H, C₆H₄-H2), 8.58 (s, 1H, C₂N₃H); ¹³C NMR

(125 MHz, CD₃OD): d 62.6 (C-6), 68.7 (C-4), 70.1 (C-2), 72.6 (C-

3), 78.7 (C-5), 88.6 (C-1), 122.7 (C₂N₃H–C5), 125.0, 126.6, 129.4,

131.6, 133.5, 136.0 (C₆H₄), 147.7 (C₂N₃H–C4); HR-MS Calcd for

4.1.7.25. 1-(

1,2,3-triazole (8o).

according to general procedure D. Yield: 25 mg, 87%.

+33.8 (c 1.12, MeOH); ¹H NMR (500 MHz, CD₃OD): d 2.35

a-

D

-Mannopyranosyl)methyl-4-(4-methylphenyl)-

Prepared from 25o (42 mg, 84 mol)

l

[a]

D

(PhCH₃), 3.72–3.76 (m, 2H, H-4, H-6a), 3.80 (dt, J = 3.2, 7.1 Hz,

1H, H-5), 3.81–3.85 (m, 2H, H-2, H-3), 3.87 (dd, J = 7.1, 11.5 Hz,

C

₁₄H₁₆ClN₃NaO₅[M+Na]⁺: 364.0676, Found: 364.0676.

O. Schwardt et al. / Bioorg. Med. Chem. 19 (2011) 6454–6473

6471

1H, H-6b), 4.24 (dt, J = 5.1, 7.4 Hz, 1H, H-1), 4.72–4.75 (m, 2H, H-

1⁰), 7.24, 7.69 (AA⁰, BB⁰of AA⁰BB⁰, J = 8.0 Hz, 4H, C₆H₄), 8.40 (s, 1H,

78.4 (C-5), 115.9 (2C, C₆H₅–C2, C6), 122.2 (C₆H₅–C4), 126.4

(C₂N₃H–C5), 130.5 (2C, C₆H₅–C3, C5), 145.0 (C₂N₃H–C4), 159.8

(C₆H₅–C1); HR-MS Calcd for C₁₆H₂₁N₃NaO₅[M+Na]⁺: 374.1328,

Found: 374.1328.

13

C₂N₃H); C NMR (125 MHz, CD₃OD): d 21.3 (PhCH₃), 50.8 (C-1⁰),

62.0 (C-6), 69.1 (C-2), 69.9 (C-4), 72.4 (C-3), 75.0 (C-1), 78.4 (C-

5), 123.0 (C₂N₃H–C5), 126.6, 128.9, 130.5, 139.3 (6C, C₆H₅), 148.9

(C₂N₃H–C4); HR-MS Calcd for C₁₆H₂₁NaN₃O₅[M+Na]⁺: 358.1379,

Found: 358.1380.

4.2. Biological evaluation

4.2.1. Competitive binding assay

4.1.7.26. 4-(3-Chlorophenyl)-1-(

1,2,3-triazole (8p). Prepared from 25p (40 mg, 77

according to general procedure D. Yield: 23 mg, 83%.

+31.5 (c 1.05, MeOH); ¹H NMR (500 MHz, CD₃OD): d 3.73

a

-

D

-mannopyranosyl)methyl-

A recombinant protein consisting of the CRD of FimH linked

with a thrombin cleavage site to a 6His-tag (FimH-CRD-Th-6His)

was expressed in E. coli strain HM125 and puriﬁed by afﬁnity chro-

matography.⁴⁵To determine the afﬁnity of the various FimH antag-

onists, an competitive binding assay described previously⁴⁵was

applied. Microtiter plates (F96 MaxiSorp, Nunc) were coated with

100 lL/well of a 10 lg/mL solution of FimH-CRD-Th-6His in

20 mM HEPES, 150 mM NaCl and 1 mM CaCl₂, pH 7.4 (assay buffer)

overnight at 4 °C. The coating solution was discarded and the wells

l

mol)

[a]

D

(m, 1H, H-4), 3.74 (dd, J = 3.0, 11.5 Hz, 1H, H-6a), 3.79–3.82 (m,

2H, H-3, H-5), 3.83 (dd, J = 3.4, 8.7 Hz, 1H, H-2), 3.89 (dd, J = 7.4,

11.6 Hz, 1H, H-6b), 4.24 (dt, J = 4.7, 7.9 Hz, 1H, H-1), 4.73 (dd,

J = 8.0, 14.4 Hz, 1H, H-1⁰a), 4.77 (dd, J = 4.4, 14.5 Hz, 1H, H-1⁰b),

7.33 (dd, J = 0.9, 8.1 Hz, 1H, C₆H₄-H6), 7.41 (t, J = 7.9 Hz, 1H,

C₆H₄–H5), 7.74 (d, J = 7.8 Hz, 1H, C₆H₄–H4), 7.90 (t, J = 1.6 Hz, 1H,

C₆H₄-H2), 8.51 (s, 1H, C₂N₃H); ¹³C NMR (125 MHz, CD₃OD): d

50.9 (C-1⁰), 62.0 (C-6), 69.0 (C-2), 70.0 (C-4), 72.4 (C-3), 74.7 (C-

1), 78.5 (C-5), 124.0 (C₂N₃H–C5), 124.9, 126.5, 129.1, 131.5,

133.9, 135.9 (6C, C₆H₄), 147.4 (C₂N₃H–C4); HR-MS Calcd for

C₁₅H₁₈ClNaN₃O₅[M+Na]⁺: 378.0833, Found: 378.0833.

were blocked with 150

4 °C. After three washing steps with assay buffer (150

four-fold serial dilution of the test compound (50 L/well) in assay

buffer containing 5% DMSO and streptavidin-peroxidase coupled

TM-PAA polymer (50 L/well of a 0.5 g/mL solution) were added.

On each individual microtiter plate n-heptyl -mannopyranoside

lL/well of 3% BSA in assay buffer for 2 h at

lL/well), a

l

a

-D

(1b) was tested in parallel. The plates were incubated for 3 h at

4.1.7.27.

syl)methyl-1,2,3-triazole (8q).

84 mol) according to general procedure D. Yield: 28 mg, 86%.

+32.6 (c 1.03, MeOH); ¹H NMR (500 MHz, CD₃OD): d 3.73–

4-(4-Triﬂuoromethylphenyl)-1-(

a-D-mannopyrano-

25 °C and 350 rpm and then carefully washed four times with

150 lL/well assay buffer. After the addition of 100 lL/well of

Prepared from 25q (47 mg,

l

[a]

ABTS-substrate, the colorimetric reaction was allowed to develop

for 4 min, then stopped by the addition of 2% aqueous oxalic acid

before the optical density (OD) was measured at 415 nm on a

microplate-reader (Spectramax 190, Molecular Devices, California,

USA). The IC₅₀values of the compounds tested in duplicates were

calculated with prism software (GraphPad Software, Inc., La Jolla,

USA). The IC₅₀deﬁnes the molar concentration of the test com-

pound that reduces the maximal speciﬁc binding of TM-PAA poly-

mer to FimH-CRD by 50%. The relative IC₅₀(rIC₅₀) is the ratio of the

IC₅₀of the test compound to the IC₅₀of 1b.

D

3.76 (m, 2H, H-4, H-6a), 3.81–3.86 (m, 3H, H-2, H-3, H-5), 3.89 (dd,

J = 7.5, 11.5 Hz, 1H, H-6b), 4.25 (dt, J = 4.8, 7.9 Hz, 1H, H-1), 4.76

(dd, J = 8.0, 14.5 Hz, 1H, H-1⁰a), 4.79 (dd, J = 4.4, 14.5 Hz, 1H, H-

1⁰b), 7.72, 8.01 (AA⁰, BB⁰of AA⁰BB⁰, J = 8.2 Hz, 4H, C₆H₄), 8.60 (s,

13

1H, C₂N₃H); C NMR (125 MHz, CD₃OD): d 51.0 (C-1⁰), 62.0 (C-6),

69.0 (C-2), 70.0 (C-4), 72.4 (C-3), 74.7 (C-1), 78.6 (C-5), 124.5

(C₂N₃H–C5), 125.6 (q, J = 271 Hz, CF₃), 126.9 (q, J = 3.7 Hz, 2C,

C₆H₄–C3, C5), 127.0 (2C, C₆H₄–C2, C6), 130.9 (q, J = 32.4 Hz,

C₆H₄–C4), 135.7 (C₆H₄–C1), 147.2 (C₂N₃H–C4); HR-MS Calcd for

C

₁₆H₁₈F₃NaN₃O₅[M+Na]⁺: 412.1096, Found: 412.1095.

4.2.2. Aggregometry assay

The aggregometry assay was carried out as previously de-

scribed.⁴⁶In short, the percentage of aggregation of E. coli

UTI89⁵⁹(UTI89wt) with guinea pig erythrocytes (GPE) was quanti-

tatively determined by measuring the optical density at 740 nm

and 37 °C under stirring at 1000 rpm using an APACT 4004 aggre-

gometer (Endotell AG, Allschwil, Switzerland). GPE were separated

from guinea pig blood (Charles River Laboratories, Sulzfeld, Ger-

many) using Histopaque (density of 1.077 g/mL at 24 °C, Sigma-Al-

drich, Buchs, Switzerland). Prior to the measurements, the cell

densities of E. coli and GPE were adjusted to an OD₆₀₀of 4, corre-

sponding to 1.9 ꢂ 10⁸CFU/mL and 2.2 ꢂ 10⁶cells/mL respectively.

For the calibration of the instrument, the aggregation of protein

poor plasma (PPP) using PBS alone was set as 100% and the aggre-

gation of protein rich plasma (PRP) using GPE as 0%. After calibra-

4.1.7.28. 1-(

azole (8r).

a

-

D

-Mannopyranosyl)methyl-4-(3-pyridyl)-1,2,3-tri-

Prepared from 25r (44 mg, 90 mol) according to

l

general procedure D. Yield: 24 mg, 83%.

[a]

D

+31.2 (c 0.99, MeOH); ¹H NMR (500 MHz, CD₃OD): d 3.71–

3.74 (m, 2H, H-4, H-6a), 3.80–3.83 (m, 3H, H-2, H-3, H-5), 3.89 (dd,

J = 7.7, 11.6 Hz, 1H, H-6b), 4.23 (dt, J = 4.6, 8.6 Hz, 1H, H-1), 4.76

(dd, J = 8.4, 14.4 Hz, 1H, H-1⁰a), 4.80 (dd, J = 4.2, 14.4 Hz, 1H, H-

1⁰b), 7.53 (dd, J = 5.0, 7.9 Hz, 1H, C₅H₄N-H5), 8.28 (d, J = 8.0 Hz,

1H, C₅H₄N-H6), 8.51 (d, J = 4.8 Hz, 1H, C₅H₄N-H4), 8.63 (s, 1H,

C₂N₃H), 9.02 (s, 1H, C₅H₄N–H2); ¹³C NMR (125 MHz, CD₃OD): d

48.1 (C-1⁰), 60.6 (C-6), 67.2 (C-2), 68.5 (C-4), 70.6 (C-3), 75.1 (C-

1), 76.2 (C-5), 123.2 (C₂N₃H–C5), 124.5 (C₅H₄N–C5), 126.82

(C₅H₄N–C1), 134.3 (C₅H₄N–C6), 144.4 (C₂N₃H–C4), 145.7 (C₅H₄N–

C2), 148.4 (C₅H₄N–C4); HR-MS Calcd for C₁₄H₁₈NaN₄O₅[M+Na]⁺:

345.1175, Found: 345.1175.

tion, measurements were performed with 250

bacterial suspension and the aggregation monitored over 600 s.

After the aggregation phase of 600 s, 25 L of antagonist in PBS

were added to each cuvette and disaggregation was monitored

for 1400 s. UTI89 ﬁmA-H was used as negative control.

lL GPE and 50 lL

l

4.1.7.29.

1,2,3-triazole (8s).

according to general procedure D. Yield: 23 mg, 83%.

+22.8 (c 1.01, MeOH); ¹H NMR (500 MHz, CD₃OD): d 3.69–

1-(

a-D

-Mannopyranosyl)methyl-4-phenoxymethyl-

Prepared from 25s (41 mg, 79 mol)

l

D

[

a]

D

4.3. Determination of the pharmacokinetic parameters

3.76 (m, 3H, H-4, H-5, H-6a), 3.79–3.82 (m, 2H, H-2, H-3), 3.83 (dd,

J = 6.5, 11.5 Hz, 1H, H-6b), 4.19 (dt, J = 5.0, 7.0 Hz, 1H, H-1), 4.69

(dd, J = 7.5, 14.5 Hz, 1H, H-1⁰a), 4.72 (dd, J = 5.0, 14.5 Hz, 1H, H-

1⁰b), 5.15 (s, 2H, CH₂OPh), 6.94 (t, J = 7.4 Hz, 1H, C₆H₅-H4), 7.00

(d, J = 8.1 Hz, 2H, C₆H₅-H2, H6), 7.27 (m, 2H, C₆H₅-H3, H5), 8.17

4.3.1. Materials

Dimethyl sulfoxide (DMSO) and 1-octanol were purchased from

Sigma-Aldrich (St. Louis MI, USA). PAMPA System Solution, GIT-0

Lipid Solution, and Acceptor Sink Buffer were ordered from pIon

(Woburn MA, USA). Acetonitrile (MeCN) was bought from Acros

Organics (Geel, Belgium).

13

(s, 1H, C₂N₃H); C NMR (125 MHz, CD₃OD): d 50.9 (C-1⁰), 62.0

(C-6), 62.3 (CH₂OPh), 69.0 (C-2), 69.8 (C-4), 72.4 (C-3), 74.9 (C-1),

6472

O. Schwardt et al. / Bioorg. Med. Chem. 19 (2011) 6454–6473

4.3.2. LC–MS measurements

performed at three pH values (5.0, 6.2 and 7.4) in quadruplicates.

For this purpose, 12 wells of a deep well plate, that is, four wells

Analyses were performed using an Agilent 1100/1200 Series

HPLC System coupled to a 6410 Triple Quadrupole mass detector

(Agilent Technologies, Inc., Santa Clara CA, USA) equipped with elec-

trospray ionization. The system was controlled with the Agilent

MassHunter Workstation Data Acquisition software (version

B.01.04). The column used was an Atlantis^ÒT3 C18 column (2.1 x

per pH-value, were ﬁlled with 650

lL PAMPA System Solution.

Samples (150 L) were withdrawn from each well to determine

l

the blank spectra by UV-spectroscopy (SpectraMax 190, Molecular

Devices, Silicon Valley Ca, USA). Then, analyte dissolved in DMSO

was added to the remaining PAMPA System Solution to yield

50 m) with a 3

lm particle size (Waters Corp., Milford MA, USA).

50 lM solutions. To exclude precipitation, the optical density

The mobile phase consisted of two eluents: solvent A (H₂O, contain-

ing 0.1% formic acid, v/v) and solvent B (MeCN, containing 0.1% for-

mic acid, v/v), both delivered at 0.6 mL/min. The gradient was

ramped from 95% A/5% B to 5% A/95% B over 1 min, and then hold

at 5% A/95% B for 0.1 min. The system was then brought back to

95% A/5% B, resulting in a total duration of 4 min. MS parameters

such as fragmentor voltage, collision energy and polarity were opti-

mized individually for each compound, and the molecular ion was

followed for each compound in the multiple reaction monitoring

mode. Theconcentrationsofthe analyteswere quantiﬁed by the Agi-

lent Mass Hunter Quantitative Analysis software (version B.01.04).

was measured at 650 nm, with 0.01 being the threshold value.

Solutions exceeding this threshold were ﬁltrated. Afterwards, sam-

ples (150

Further 200

l

L) were withdrawn to determine the reference spectra.

L were transferred to each well of the donor plate of

l

the PAMPA sandwich (pIon, Woburn MA, USA, P/N 110 163). The

ﬁlter membranes at the bottom of the acceptor plate were impreg-

nated with 5 lL of GIT-0 Lipid Solution and 200 lL of Acceptor Sink

Buffer were ﬁlled into each acceptor well. The sandwich was

assembled, placed in the GutBox™, and left undisturbed for 16 h.

Then, it was disassembled and samples (150 lL) were transferred

from each donor and acceptor well to UV-plates. Quantiﬁcation

was performed by both UV-spectroscopy and LC-MS. log P_e-values

were calculated with the aid of the PAMPA Explorer Software

(pIon, version 3.5).

4.3.3. log D_7.4determination

The in silico prediction tool ALOGPS⁶⁰was used to estimate the

log P values. Depending on these values, the compounds were clas-

siﬁed into three categories: hydrophilic compounds (log P below

zero), moderately lipophilic compounds (log P between zero and

one) and lipophilic compounds (log P above one). For each cate-

gory, two different ratios (volume of 1-octanol to volume of buffer)

were deﬁned as experimental parameters:

4.3.5. Thermodynamic solubility

Microanalysis tubes (Labo-Tech J. Stofer LTS AG, Muttenz, Swit-

zerland) were charged with 1 mg of solid substance and 250 lL of

phosphate buffer (50 mM, pH 6.5). The samples were brieﬂy sha-

ken by hand, then sonicated for 15 min and vigorously shaken

(600 rpm, 25 °C, 2 h) on a Eppendorf Thermomixer comfort. After-

wards, the samples were left undisturbed for 24 h. After measuring

the pH, the saturated solutions were ﬁltered through a ﬁltration

plate (MultiScreen^ÒHTS, Millipore, Billerica MA, USA) by centrifu-

gation (1500 rpm, 25 °C, 3 min). Prior to concentration determina-

tion by LC-MS, the ﬁltrates were diluted (1:1, 1:10 and 1:100 or, if

the results were outside of the calibration range, 1:1000 and

1:10000). The calibration was based on six values ranging from

Compound type

log P

Ratios (1-octanol:buffer)

Hydrophilic

Moderately lipophilic

Lipophilic

<0

0–1

>1

30:140, 40:130

70:110, 110:70

3:180, 4:180

Equal amounts of phosphate buffer (0.1 M, pH 7.4) and 1-octa-

nol were mixed and shaken vigorously for 5 min to saturate the

phases. The mixture was left until separation of the two phases oc-

curred, and the buffer was retrieved. Stock solutions of the test

0.1 to 10 lg/mL.

Acknowledgement

compounds were diluted with buffer to a concentration of 1 lM.

For each compound, six determinations, that is, three determina-

tions per 1-octanol : buffer ratio, were performed in different wells

of a 96-well plate. The respective volumes of buffer containing ana-

We thank the Swiss National Science Foundation (project K-

32KI-120904) for their support.

lyte (1 lM) were pipetted to the wells and covered by saturated 1-

Supplementary data

octanol according to the chosen volume ratio. The plate was sealed

with aluminium foil, shaken (1350 rpm, 25 °C, 2 h) on a Heidoph

Titramax 1000 plate-shaker (Heidolph Instruments GmbH & Co.

KG, Schwabach, Germany) and centrifuged (2000 rpm, 25 °C,

5 min, 5804 R Eppendorf centrifuge, Hamburg, Germany). The

aqueous phase was transferred to a 96-well plate for analysis by li-

quid chromatography-mass spectrometry (LC-MS).

Supplementary data (HRMS and HPLC data of target compounds

5–8) associated with this article can be found, in the online version,

at doi:10.1016/j.bmc.2011.08.057.

References and notes

log D_7.4was calculated from the 1-octanol : buffer ratio (o:b),

1. Wiles, T. J.; Kulesus, R. R.; Mulvey, M. A. Exp. Mol. Pathol. 2008, 85, 11.

2. Fihn, S. D. N. Engl. J. Med. 2003, 349, 259.

3. Hooton, T. M. Int. J. Antimicrob. Agents 2001, 17, 259.

4. Gouin, S. G.; Wellens, A.; Bouckaert, J.; Kovensky, J. ChemMedChem 2009, 4, 749.

5. Rosen, D. A.; Hung, C. S.; Kline, K. A.; Hultgren, S. J. Infect. Immun. 2008, 76,

4290.

the initial concentration of the analyte in buffer (1

lM), and the

concentration of the analyte in buffer (c_B) with equilibration:

ꢀ

ꢁ

1

l

M ꢁ c_B

1

o : b

log D_7:4

¼

ꢂ

c_B

6. Capitani, G.; Eidam, O.; Glockshuber, R.; Grutter, M. G. Microbes Infect. 2006, 8,

2284.

The average of the three log D_7.4values per 1-octanol:buffer ra-

tio was calculated. If the two mean values obtained for a com-

pound did not differ by more than 0.1 unit, the results were

accepted.

7. Mulvey, M. A. Cell Microbiol. 2002, 4, 257.

8. Choudhury, D.; Thompson, A.; Stojanoff, V.; Langermann, S.; Pinkner, J.;

Hultgren, S. J.; Knight, S. D. Science 1999, 285, 1061.

9. Bouckaert, J.; Berglund, J.; Schembri, M.; Genst, E. D.; Cools, L.; Wuhrer, M.;

Hung, C. S.; Pinkner, J.; Slättergård, R.; Zavialov, A.; Choudhury, D.;

Langermann, S.; Hultgren, S. J.; Wyns, L.; Klemm, P.; Oscarson, S.; Knight, S.

D.; Greve, H. D. Mol. Microbiol. 2005, 55, 441.

10. Wellens, A.; Garofalo, C.; Nguyen, H.; Van Gerven, N.; Slättergård, R.;

Hernalsteens, J.-P.; Wyns, L.; Oscarson, S.; De Greve, H.; Hultgren, S.;

Bouckaert, J. PLoS ONE 2008, 3, 4.

4.3.4. Parallel artiﬁcial membrane permeation assay (PAMPA)

log P_ewas determined in a 96-well format with the PAMPA⁵⁰

permeation assay. For each compound, measurements were

O. Schwardt et al. / Bioorg. Med. Chem. 19 (2011) 6454–6473

6473

11. Han, Z.; Pinker, J. S.; Ford, B.; Obermann, R.; Nolan, W.; Wildman, S. A.; Hobbs,

D.; Ellenberger, T.; Cusumano, C. K.; Hultgren, S. J.; Janetka, J. W. J. Med. Chem.

2010, 53, 4779.

12. Langermann, S.; Mollby, R.; Burlein, J. E.; Palaszynski, S. R.; Auguste, C. G.;

DeFusco, A.; Strouse, R.; Schenerman, M. A.; Hultgren, S. J.; Pinkner, J. S.;

Winberg, J.; Guldevall, L.; Soderhall, M.; Ishikawa, K.; Normark, S.; Koenig, S. J.

Infect. Dis. 2000, 181, 774.

13. Langermann, S.; Palaszynski, S.; Barnhart, M.; Auguste, G.; Pinkner, J. S.;

Burlein, J.; Barren, P.; Koenig, S.; Leath, S.; Jones, C. H.; Hultgren, S. J. Science

1997, 276, 607.

33. (a) Tornøe, C. W.; Meldal, M. In Peptides: The Wave of the Future: Proceedings of

the Second International and the Seventeenth American Peptide Symposium; Lebl,

M., Houghten, R. A., Eds.; Springer, 2001; p 263; (b) Tornøe, C. W.; Christensen,

C.; Meldal, M. J. Org. Chem. 2002, 67, 3057.

34. Kolb, H. C.; Sharpless, K. B. Angew. Chem., Int. Ed. 2001, 40, 2004.

35. Palomo, C.; Aizpurua, J. M.; Balentová, E.; Azcune, I.; Santos, J. I.; Jiménez-

Barbero, J.; Cañada, J.; Miranda, J. I. Org. Lett. 2008, 10, 2227.

36. (a) Tietze, L.; Bothe, U. Chem. Eur. J. 1998, 4, 1179; (b) Gouin, S. G.; Vanquelef,

E.; García Fernández, J. M.; Ortiz Mellet, C.; Dupradeau, F.-Y.; Kovensky, J. J. Org.

Chem. 2007, 72, 9032.

14. Bouckaert, J.; Mackenzie, J.; de Paz, J. L.; Chipwaza, B.; Choudhury, D.; Zavialov,

A.; Mannerstedt, K.; Anderson, J.; Pierard, D.; Wyns, L.; Seeberger, P. H.;

Oscarson, S.; De Greve, H.; Knight, S. D. Mol. Microbiol. 2006, 61, 1556.

15. (a) Sharon, N. Biochim. Biophys. Acta 2006, 1760, 527; (b) Ofek, I.; Hasty, D. L.;

Sharon, N. FEMS Immun. Med. Microbiol. 2003, 38, 181; (c) Sharon, N.; Ofek, I. In

Protein-Carbohydrate Interactions in Infectious Diseases; Bewley, C. A., Ed.; RSC

Biomolecular Sciences: Cambridge, UK, 2006; pp 49–72.

37. Fernández-Megia, E.; Correa, J.; Rodríguez-Meizoso, I.; Riguera, R.

Macromolecules 2006, 39, 2113.

38. Buckle, D. R.; Rockell, C. J. M. J. Chem. Soc., Perkin Trans. I 1982, 627.

39. Stefely, J. A.; Palchaudhuri, R.; Miller, P. A.; Peterson, R. J.; Moraski, G. C.;

Hergenrother, P. J.; Miller, M. J. J. Med. Chem. 2010, 53, 3389.

40. Benati, L.; Bencivenni, G.; Leardini, R.; Minozzi, M.; Nanni, D.; Scialpi, R.;

Spagnolo, P.; Zanardi, G. J. Org. Chem. 2006, 71, 5822.

16. (a) Firon, N.; Ofek, I.; Sharon, N. Biochem. Biophys. Res. Commun. 1982, 105,

1426; (b) Firon, N.; Ofek, I.; Sharon, N. Carbohydr. Res. 1983, 120, 235; (c)

Sharon, N. FEBS Lett. 1987, 217, 145.

41. Ito, S.; Satoh, A.; Nagatomi, Y.; Hirata, Y.; Suzuki, G.; Kimura, T.; Satow, A.;

Maehara, S.; Hikichi, H.; Hata, M.; Kawamoto, H.; Ohta, H. Bioorg. Med. Chem.

2008, 16, 9817.

17. (a) Neeser, J.-R.; Koellreutter, B.; Wuersch, P. Infect. Immun. 1986, 52, 428; (b)

Lindhorst, T. K. Top. Curr. Chem. 2002, 218, 201. review; (c) Patel, A.; Lindhorst,

T. K. Carbohydr. Res. 2006, 341, 1657; (d) Nagahori, N.; Lee, R. T.; Nishimura, S.-

L.; Pagé, S.; Roy, R.; Lee, Y. C. ChemBioChem 2002, 3, 836; (e) Appeldoorn, C. C.

M.; Joosten, J. A. F.; Maate, F. A.; Dobrindt, U.; Hacker, J.; Liskamp, R. M. J.; Khan,

A. S.; Pieters, R. J. Tetrahedron: Asymmetry 2005, 16, 361; (f) Touaibia, M.;

Wellens, A.; Shiao, T. C.; Wang, Q.; Sirois, S.; Bouckaert, J.; Roy, R.

ChemMedChem 2007, 2, 1190.

18. (a) Firon, N.; Ashkenazi, S.; Mirelman, D.; Ofek, I.; Sharon, N. Infect. Immun.

1987, 55, 472; (b) Lindhorst, T. K.; Kötter, S.; Kubisch, J.; Krallmann-Wenzel, U.;

Ehlers, S.; Kren, V. Eur. J. Org. Chem. 1998, 1669; (c) Sperling, O.; Fuchs, A.;

Lindhorst, T. K. Org. Biomol. Chem. 2006, 4, 3913; (d) Berglund, J.; Bouckaert, J.;

De Greve, H.; Knight, S. PCT/US 2005/089733, 2005.

19. Klein, T.; Abgottspon, D.; Wittwer, M.; Rabbani, S.; Herold, J.; Jiang, X.; Kleeb,

S.; Lüthi, C.; Scharenberg, M.; Bezençon, J.; Gubler, E.; Pang, L.; Smiesko, M.;

Cutting, B.; Schwardt, O.; Ernst, B. J. Med. Chem. 2010, 53, 8627.

20. Hung, C. S.; Bouckaert, J.; Hung, D.; Pinkner, J.; Widberg, C.; Defusco, A.;

Auguste, C. G.; Strouse, R.; Langermann, S.; Waksman, G.; Hultgren, S. J. Mol.

Microbiol. 2002, 44, 903.

42. (a) Kunz, H.; Pfrengle, W.; Rück, K.; Sager, W. Synthesis 1991, 103, 1039; (b)

Györgydeak, Z.; Szilagyi, L.; Paulsen, H. J. Carbohydr. Chem. 1993, 12, 139.

43. (a) Myers, R. W.; Lee, Y. C. Carbohydr. Res. 1986, 154, 145; (b) Köll, P.; Förtsch, A.

Carbohydr. Res. 1987, 171, 301.

44. Titz, A.; Radic, Z.; Schwardt, O.; Ernst, B. Tetrahedron Lett. 2006, 47, 2383.

45. Rabbani, S.; Jiang, X.; Schwardt, O.; Ernst, B. Anal. Biochem. 2010, 407, 188.

46. Abgottspon, D.; Rölli, G.; Hosch, L.; Steinhuber, A.; Jiang, X.; Schwardt, O.;

Cutting, B.; Smiesko, M.; Jenal, U.; Ernst, B.; Trampuz, A. J. Microbiol. Methods

2010, 82, 249.

47. Giampapa, C. S.; Abraham, S. N.; Chiang, T. M.; Beachey, E. H. J. Biol. Chem. 1988,

263, 5362.

48. Aprikian, P.; Tchesnokova, V.; Kidd, B.; Yakovenko, O.; Yarov-Yarovoy, V.;

Trinchina, E.; Vogel, V.; Thomas, W.; Sokurenko, E. J. Biol. Chem. 2007, 282,

23437.

49. Trong, I. L.; Aprikian, P.; Kidd, B. A.; Forero-Shelton, M.; Tchesnokova, V.;

Rajagopal, P.; Rodriguez, V.; Interlandi, G.; Klevit, R.; Vogel, V.; Stenkamp, R. E.;

Sokurenko, E. V.; Thomas, W. E. Cell 2010, 141, 645.

50. Kansy, M.; Senner, F.; Gubernator, K. J. Med. Chem. 1998, 41, 1007.

51. Dearden, J. C.; Bresnen, J. G. M. QSAR Comb. Sci. 1988, 7, 133.

52. Kerns, E. H. J. Pharm. Sci. 2001, 90, 1838.

53. Poulain, F.; Serre, A.-L.; Lalot, J.; Leclerc, E.; Quirion, J.-C. J. Org. Chem. 2008, 73,

2435.

54. Kessler, H.; Gehrke, M.; Griesinger, C. Angew. Chem. 1988, 100, 507; Angew.

Chem. Int. Ed. 1988, 27, 490.

21. Ernst, B.; Magnani, J. L. Nat. Rev. Drug Disc. 2009, 8, 661.

22. Becke, A. D. J. Chem. Phys. 1993, 98, 5648.

23. Stephens, P. J.; Devlin, F. J.; Chabalowski, C. F.; Frisch, M. J. J. Phys. Chem. 1994,

98, 11623.

24. Frisch, M. J.; Trucks, G. W.; Schlegel, H. B.; Scuseria, G. E.; Robb, M. A.;

Cheeseman, J. R.; Montgomery, J., J.A.; Vreven, T.; Kudin, T. K. N.; Burant, J. C.

et al. Gaussian 2003, Gaussian Inc: Pittsburgh, PA, 2003.

55. Reid, B. R.; Banks, K.; Flynn, P.; Nerdalt, W. Biochemistry 1989, 28, 10001.

56. Szilágyi, L.; Forgó, P. Carbohydr. Res. 1993, 247, 129.

25. Sancho-Garcia, J. C.; Cornil, J. J. Chem. Theory Comput. 2005, 1, 581.

26. Glide, version 5.5; Schrödinger, LLC: New York, NY, 2009.

27. Desmond Molecular Dynamics System, version 2.2; D. E. Shaw Research: New

York, NY, 2009.

57. Varma, M. V. S.; Feng, B.; Obach, R. S.; Troutman, M. D.; Chupka, J.; Miller, H. R.;

El-Kattan, A. J. Med. Chem. 2009, 52, 4844.

58. Avdeef, A.; Bendels, S.; Di, L.; Faller, B.; Kansy, M.; Sugano, K.; Yamauchi, Y. J.

Pharm. Sci. 2007, 96, 2893.

28. Humphrey, W.; Dalke, A.; Schulten, K. J. Mol. Graphics 1996, 14, 33.

29. Kaufman, R. J.; Sidhu, R. S. J. Org. Chem. 1982, 47, 4941.

30. (a) Lamara, K.; Smalley, R. K. Tetrahedron 1991, 47, 2277; (b) Li, Y.; Gao, L.-X.;

Han, F.-S. Chem. Eur. J. 2010, 16, 7969.

59. Mulvey, M. A.; Schilling, J. D.; Hultgren, S. J. Infect. Immun. 2001, 69, 4572.

60. VCCLAB, Virtual Computational Chemistry Laboratory, http://www.vcclab.org,

2005.; (b) Tetko, I. V.; Gasteiger, J.; Todeschini, R.; Mauri, A.; Livingstone, D.;

Ertl, P.; Palyulin, V. A.; Radchenko, E. V.; Zeﬁrov, N. S.; Makarenko, A. S.;

Tanchuk, V. Y.; Prokopenko, V. V. J. Comput. Aid. Mol. Des. 2005, 19, 453.

31. Hu, M.; Li, J.; Yao, S. Q. Org. Lett. 2008, 10, 5529.

32. (a) Sawanishi, H.; Tajima, K.; Tsuchiya, T. Chem. Pharm. Bull. 1987, 35, 4101; (b)

Sapountzis, I.; Ettmayer, P.; Klein, C.; Mantoulidis, A.; Steegmaier, M.; Steurer,

S.; Waizenegger, I. PCT/EP2008/058432, 2008.

Article Doi

Design, synthesis and biological evaluation of mannosyl triazoles as FimH antagonists

DOI: 10.1016/j.bmc.2011.08.057

Source and publish data:

Authors:

Article abstract of DOI:10.1016/j.bmc.2011.08.057

Full text of DOI:10.1016/j.bmc.2011.08.057

Products guided by the article

R&D Labs maybe for 1345018-99-6

Relevant to this article

Hot Product