2010-Glycosidase Inhibition With Fullerene Iminosugar Balls a Dramatic Multivalent Effect-SI

Supporting Information

� Wiley-VCH 2010

69451 Weinheim, Germany

Glycosidase Inhibition with Fullerene Iminosugar Balls: A DramaticMultivalent Effect**Philippe Compain,* Camille Decroocq, Julien Iehl, Michel Holler, Damien Hazelard,Teresa Mena Barrag�n, Carmen Ortiz Mellet,* and Jean-Fran�ois Nierengarten*

anie_201002802_sm_miscellaneous_information.pdf

S1

Supplementary Information

S2

Table of Contents

General Methods S3

Syntheses and Analytical data of the Compounds S3 1H and 13C NMR Spectra of the Compounds S10

MALDI-TOF-MS of compound 9 S16

UV/vis spectra of 8 and 9 S17

General Procedures for Inhibition Assay S18

References S24

S3

Experimental section

General Methods

Tetrahydrofuran (THF) was distilled over sodium/benzophenone under Ar. Dichloromethane

(CH2Cl2) was distilled over CaH2 under Ar. Dimethylformamide (DMF) was distilled over

MgSO4 under reduced pressure. Triethylamine (Et3N) was distilled over KOH under Ar and

stored over KOH. All reactions were performed in standard glassware under Ar. Column

chromatography: silica gel 60 (230-400 mesh, 0.040-0.063 mm) was purchased from E.

Merck. Thin Layer Chromatography (TLC) was performed on aluminum sheets coated with

silica gel 60 F254 purchased from E. Merck. IR spectra (cm-1) were recorded on a Perkin–

Elmer Spectrum One Spectrophotometer. NMR spectra were recorded on a Bruker AC 300 or

AC 400 with solvent peaks as reference. Carbon multiplicities were assigned by distortionless

enhancement by polarization transfer (DEPT) experiments. The 1H signals were assigned by

2D experiments (COSY). MALDI-TOF-mass spectra were carried out on a Bruker

BIFLEXTM matrix-assisted laser desorption time-of-flight mass spectrometer. ESI-HRMS

mass spectra were carried out on a Bruker MicroTOF spectrometer. Specific rotations were

determined at room temperature (20°C) in a Perkin–Elmer 241 polarimeter for sodium (λ =

589 nm).

2,3,4,6-Tetra-O-benzyl-D-gluconamide (2)

OBnO

OBn

OBn NH3 (30%), I2OH

BnO

OBn

NH2

OBn

OTHF, r.t. overnight

BnO BnO

OH1 2

A 30% aqueous NH3 solution (21 mL) and iodine (650 mg, 2.57 mmol) were added to a

solution of 1 (1.07 g, 1.98 mmol) in THF (4.5 mL). After 16 h, a 5% aqueous Na2S2O3

solution (3 mL) was added. The resulting mixture was extracted with Et2O (3 x 25 mL). The

combined organic layers were dried (Na2SO4), filtered and concentrated. Column

chromatography (SiO2, AcOEt/petroleum ether 2:1) gave 2 (854 mg, 78%) as an amorphous

S4

white solid. The analytical data of 2 were in complete agreement with those reported in the

literaturei: 1H NMR (300 MHz, CDCl3): δ = 2.81 (d, J = 4 Hz, 1H, O-H), 3.58 (dd, J = 9 and 5

Hz, 1H, H-6A), 3.65 (dd, J = 9 and 3 Hz, 1H, H-6B), 3.83-3.94 (m, 2H, H-4, H-5), 4.07 (dd, J

= 3 and 5 Hz, 1H, H-3), 4.25 (d, J = 3 Hz, 1H, H-2), 4.46-4.76 (m, 8H, CH2Ph), 5.43 (s, 1H,

NH), 6.60 (s, 1H, NH), 7.19-7.40 (m, 20H, ArH).

2,3,4,6-Tetra-O-benzyl-D-glucono-δ-lactam (3)

NHBnO

OBn

OBn

O

BnOOHBnO

OBn

NH2

OBn

O

BnO

2

1) DMSO, Ac2O

2) NaCNBH3, HCOOHCH3CN 3

A solution of 2 (911 mg, 1.64 mmol) and acetic anhydride (3.5 mL) in DMSO (6 mL) was

stirred at room temperature for 17 h. The mixture was then cooled at 0°C and H2O (22 mL)

was added. The mixture was stirred for another 15 min., then extracted with Et2O (3 x 30

mL). The combined organic layers were washed with brine, dried (Na2SO4), filtered and

concentrated. The product was used for the next step without further purification.

The resulting liquid was dissolved in CH3CN (35 mL) and formic acid (6.2 mL) was added.

NaCNBH3 (330 mg, 5.25 mmol) was then added and the mixture heated under reflux for 3 h.

The mixture was then cooled at 0°C and a 0.1 M aqueous HCl solution (50 mL) was added.

The resulting mixture was poured into a 1:1 mixture of ethyl acetate/saturated aqueous

NaHCO3 (100 mL). The aqueous layer was extracted with AcOEt (3 x 50 mL). The combined

organic layers were dried (Na2SO4), filtered and concentrated. Column chromatography

(SiO2, AcOEt/petroleum ether 1:1) gave 3 (532 mg, 60 % over the two steps) as a white solid.

The analytical data of 3 were in complete agreement with those reported in the literatureii: 1H

NMR (300 MHz, CDCl3): δ = 3.25 (td, J = 1.5 and 8 Hz, 1H, H-4), 3.44-3.63 (m, 3H, H-6A,

H-6B, H-5), 3.90 (t, J = 8 Hz, 1H, H-3), 4.00 (d, J = 8 Hz, 1H, H-2), 4.47 (m, 3H, CH2Ph),

4.72 (d, J = 11 Hz, 1H, CH2Ph), 4.77 (d, J = 11 Hz, 1H, CH2Ph), 4.84 (d, J = 11 Hz, 1H,

CH2Ph), 4.85 (d, J = 11 Hz, 1H, CH2Ph), 5.17 (d, J = 11 Hz, 1H, CH2Ph), 5.88 (s, 1H, NH),

7.13-7.46 (m, 20H, ArH).

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2,3,4,6-Tetra-O-benzyl-1,5-dideoxy-1,5-imino-D-glucitol (10)

NHBnO

OBn

OBn

BnONHBnO

OBn

OBn

O

BnO

3

LAH

THF, reflux10

A solution of 3 (532 mg, 0.99 mmol) in dry THF (8 mL) was added dropwise to a suspension

of LAH (113 mg, 2.97 mmol) in dry THF (6 mL) at 0°C. The reaction mixture was heated

under reflux for 2 h, then cooled at 0°C. H2O (0.12 mL) and a 15% aqueous NaOH solution

(0.12 mL) were successively added to the mixture. After, 15 min., an additional portion of

H2O (0.8 mL) was added. The resulting mixture was filtered through a pad of celite (Et2O)

and concentrated. Column chromatography (SiO2, AcOEt/petroleum ether 1:1) gave 10 (465

mg, 90%) as a colorless syrup. The analytical data of 10 were in complete agreement with

those reported in the literatureii: 1H NMR (300 MHz, CDCl3): δ = 2.51 (dd, J = 12 and 10 Hz,

1 H, H-1A), 2.73 (ddd, J = 9, 6 and 3 Hz, 1H, H-5), 3.25 (dd, J = 12 and 4.5 Hz, 1H, H-1B),

3.36 (t, J = 9 Hz, 1H, H-4), 3.46-3.60 (m, 3H, H-2, H-3, H-6A), 3.68 (dd, J = 9 and 3 Hz, 1H,

H-6B), 4.39-4.53 (m, 3H), 4.65 (d, J = 11.5 Hz, 1H, CH2Ph), 4.71 (d, J = 11.5 Hz, 1H,

CH2Ph), 4.83 (d, J = 8 Hz, 1H, CH2Ph), 4.87 (d, J = 8 Hz, 1H, CH2Ph), 4.98 (d, J = 11 Hz,

1H, CH2Ph), 7.17-7.23 (m, 2H, ArH), 7.23-7.38 (m, 18H, ArH).

N-(6-Azidohexyl)-2,3,4,6-tetra-O-benzyl-1,5-dideoxy-1,5-imino-D-glucitol (4)

NHBnO

OBn

OBn

BnO

10

BrN3

NEt3, DMAP, DMF120°C,4 days

NBnO

OBn

OBn

BnON3

4

5

A mixture of 10 (120 mg, 0.23 mmol), 1-azido-6-bromohexaneiii (236 mg, 1.15 mmol), Et3N

(0.3 mL, 2.29 mmol) and DMAP (4 mg, 0.03 mmol) in DMF (4.5 mL) was stirred at 120°C

for 4 days. The mixture was cooled at room temperature and H2O (9 mL) was added. The

aqueous layer was extracted with Et2O (3 x 25 mL). The combined organic layers were dried

(Na2SO4), filtered and concentrated. Column chromatography (SiO2, AcOEt/petroleum ether

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5:1) gave 4 (57 mg, 38%) as a yellow oil. [ ]20Dα -5 (c 1, CHCl3).

1H NMR (300 MHz, CDCl3):

δ = 1.00-1.17 (m, 2H), 1.17-1.41 (m, 4H), 1.49 (m, J = 7 Hz, 2H, H-11), 2.15 (t, J = 11 Hz,

1H, H-1A), 2.25 (br d, J = 9 Hz, 1H, H-5), 2.42-2.55 (m, 1H, H-7A), 2.55-2.69 (m, 1H, H-

7B), 3.02 (dd, J = 11 and 5 Hz, 1H, H-1B), 3.16 (t, J = 7 Hz, 2H, H-12), 3.40 (t, J = 9 Hz, 1H,

H-3), 3.45-3.54 (m, 2H), 3.54-3.65 (m, 2H), 4.37-4.44 (m, 3H, CH2Ph), 4.58 (d, J = 12 Hz,

1H, CH2Ph), 4.64 (d, J = 12 Hz, 1H, CH2Ph), 4.75 (d, J = 11 Hz, 1H, CH2Ph), 4.82 (d, J = 11

Hz, 1H, CH2Ph), 4.90 (d, J = 11 Hz, 1H, CH2Ph), 7.03-7.12 (m, 2H, ArH), 7.12-7.37 (m, 18H,

ArH); 13C NMR (75 MHz, CDCl3): δ = 23.8, 26.7, 27.1, 28.9, 51.4, 52.3, 54.6, 64.0, 65.7,

72.8, 73.5, 75.3, 75.4, 78.7, 78.7, 87.4, 127.5, 127.6, 127.7, 127.9, 128.4, 128.4, 128.5, 137.9,

138.7, 139.1; IR (neat) : 2093 (N3) cm-1; HRMS (ESI): m/z 649.378 ([M+H]+, calcd. for

C40H49N4O4: 649.375).

N-(6-Azidohexyl)-1,5-dideoxy-1,5-imino-D-glucitol (8)

NBnO

OBn

OBn

BnON3

4

5 NHO

OH

OH

HON3

5

5

BCl3

CH2Cl2 -60°C to 0°C

A 1 M BCl3 solution in CH2Cl2 (0.62 mL, 0.62 mmol) was added to a solution of 4 (154 mg,

0.24 mmol) in CH2Cl2 (3 mL) at –60°C. The resulting mixture was allowed to warm to 0°C

over 3 h. A 20:1 (v/v) MeOH/H2O mixture (3 mL) was added and the resulting mixture

concentrated under vacuum, those steps were repeated twice. MeOH was added and the

mixture filtered through an anionic resin (AMBERLITE IRA-440C). The filtrate was

concentrated. Column chromatography (SiO2, CH2Cl2/MeOH 9:1) gave 5 (53 mg, 77%) as a

colorless oil. The analytical data of 5 were in complete agreement with those reported in the

literatureiv. [ ]20Dα -13 (c 1, MeOH). 1H NMR (300 MHz, CD3OD): δ = 1.29-1.52 (m, 4H),

1.52-1.69 (m, 4H), 2.30-2.45 (m, 2H, H-1A, H-5), 2.66-2.81 (m, 1H, H-7A), 2.87-3.03 (m,

1H, H-7B), 3.12 (dd, J = 11 and 5 Hz, 1H, H-1B), 3.20 (t, J = 9 Hz, 1H, H-3), 3.28-3.35 (m,

2H, H-12), 3.42 (t, J = 9 Hz, 1H, H-4), 3.54 (td, J = 9 and 5 Hz, 1H, H-2), 3.90 (d, J = 1.5 Hz,

2H, H-6); 13C NMR (75 MHz, CD3OD): δ = 25.0, 27.6, 27.9, 29.8, 52.4, 53.7, 57.1, 58.6,

S7

67.4, 70.2, 71.4, 80.1; IR (neat) : 3356 (O-H), 2095 (N3) cm-1; HRMS (ESI): m/z 289.184

([M+H] +, calcd. for C12H25N4O4: 289.187).

N-(6-(4-Propyl-1H-1,2,3-triazol-1-yl))-1,5-dideoxy-1,5-imino-D-glucitol (6)

NHO

OH

OH

HON3

5

5

CuSO4Sodium Ascorbate

H2O/DMF 1:1

NHO

OH

OH

HON

6

5

NNH

A mixture of 5 (16 mg, 0.06 mmol), 1-pentyne (0.03 mL, 0.28 mmol), CuSO4.5H2O (1 mg,

0.006 mmol) and sodium ascorbate (3 mg, 0.02 mmol) in DMF/H2O (1:1, 2 mL) was stirred at

room temperature. After 19 h, an additional portion of 1-pentyne (0.03 mL, 0.28 mmol) was

added and the mixture was heated at 50°C for 2 h. The mixture was then filtered through a

pad of celite and concentrated. Column chromatography (SiO2, CH2Cl2/MeOH 9:1) gave 6

(12 mg, 61% not optimized) as a colorless oil. [ ]20Dα -10 (c 0.82, MeOH). 1H NMR (300 MHz,

CD3OD): δ = 0.96 (t, J = 7 Hz, 3H, H-17), 1.24-1.42 (m, 4 H, H-9, H-10), 1.45-1.59 (m, 2 H,

H-8), 1.69 (m, J = 7 Hz, 2H, H-16), 1.91 (br t, J = 7 Hz, 2H, H-11), 2.25-2.37 (m, 2H, H-1A,

H-5), 2.59-2.75 (m, 3H, H-7A, H-15), 2.82-2.97 (m, 1H, H-7B), 3.07 (dd, J = 11 and 5 Hz,

1H, H-1B), 3.17 (t, J = 9 Hz, 1H, H-3), 3.39 (t, J = 9 Hz, 1H, H-4), 3.50 (td, J = 9 and 5 Hz,

1H, H-2), 3.87 (d, J = 3 Hz, 2H, H-6), 4.37 (t, J = 7 Hz, 2H, H-12), 7.73 (s, 1H, H-13); 13C

NMR (75 MHz, CD3OD): δ = 14.0, 23.8, 25.0, 27.3, 27.7, 28.3, 31.2, 51.1, 53.7, 57.2, 58.8,

67.5, 70.2, 71.5, 80.2, 123.1, 149.1; IR (neat) : 3317 cm-1; HRMS (ESI): m/z 357.250

([M+H] +, calcd. for C17H33N4O4: 357.246).

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Compound 8.

A 1 M solution of TBAF in THF (0.19 mL, 0.19 mmol) was added to a mixture of 7v (40 mg,

0.013 mmol), 5 (52 mg, 0.17 mmol), CuSO4.5H2O (0.2 mg, 0.001 mmol) and sodium

ascorbate (0.8 mg, 0.004 mmol) in CH2Cl2/H2O/DMSO (1:1:1, 1.5 mL). The resulting

mixture was vigorously stirred at room temperature. After 24 h, methanol (10 mL) was added

to the mixture and the resulting orange precipitate filtered, extensively washed with methanol

then CH2Cl2 and dried under high vacuum to give 8 (62 mg, 83%) as a red-orange powder. IR

(neat): 3310 (O-H), 1740 (C=O); UV/Vis (H2O): 246 (sh, 93800), 270 (79900), 285 (73700),

320 (sh, 45700), 337 (sh, 36700); 1H NMR (DMSO-d6, 300 MHz): δ = 1.23 (m, 48H), 1.39

(m, 24H), 1.77 (m, 24H), 2.15 (m, 24H), 2.63 (m, 24H), 2.85 (m, 12H), 2.96 (m, 12H), 3.10

(m, 12H), 3.28 (m, 12H), 3.46 (m, 12H), 3.61 (m, 12H), 3.68 (m, 24H), 4.27 (m, 48H), 7.81

(s, 12H); 13C NMR (DMSO-d6, 100 MHz): δ = 21.3, 23.9, 25.7, 26.2, 27.6, 29.6, 45.5, 49.1,

51.9, 56.3, 58.3, 66.4, 66.5, 68.8, 70.2, 78.2, 121.7, 140.7, 144.9, 145.5, 162.7.

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Compound 9.

A 1 M solution of TBAF in THF (0.1 mL, 0.1 mmol) was added to a mixture of 7 (20 mg,

0.0066 mmol), 4 (56 mg, 0.086 mmol), CuSO4.5H2O (0.1 mg, 0.0006 mmol) and sodium

ascorbate (0.4 mg, 0.002 mmol) in CH2Cl2/H2O (1:1, 0.5 mL). The resulting mixture was

vigorously stirred at room temperature. After 24 h, the organic layer was diluted with CH2Cl2,

washed with water, dried (MgSO4) and concentrated. Column chromatography (SiO2, CH2Cl2

containing 2% of methanol) followed by gel permeation chromatography (Biobeads SX-1,

CH2Cl2) gave 9 (52 mg, 78%) as an orange glassy product. IR (neat): 1742 (C=O); UV/Vis

(CH2Cl2): 247 (sh, 110400), 258 (86100), 265 (82000), 269 (80700), 283 (72300), 320 (sh,

39400), 339 (sh, 27900); 1H NMR (300 MHz, CDCl3): δ = 1.10-1.40 (m, 72H), 1.82 (m,

24H), 2.09 (m, 24H), 2.18 (t, J = 10 Hz, 12H), 2.27 (br d, J = 9 Hz, 12H), 2.52 (m, 12H), 2.64

(m, 12H), 2.76 (m, 24H), 3.06 (dd, J = 11 and 5 Hz, 12H), 3.44 (t, J = 9 Hz, 12H), 3.53 (m,

24H), 3.62 (m, 24H), 4.23 (m, 24H), 4.34 (m, 24H), 4.44 (m, 36H), 4.63 (d, J = 11 Hz, 12H),

4.67 (d, J = 11 Hz, 12H), 4.79 (d, J = 11 Hz, 12H), 4.86 (d, J = 11 Hz, 12H), 4.94 (d, J = 11

Hz, 12H), 7.03-7.12 (m, 24H), 7.12-7.37 (m, 206H); 13C NMR (CDCl3, 100 MHz): δ = 22.1,

23.6, 26.4, 26.9, 28.1, 29.6, 30.3, 45.4, 50.0, 52.2, 54.4, 63.8, 65.5, 66.3, 69.1, 72.7, 73.3,

75.1, 75.2, 78.4, 78.5, 87.3, 121.0, 127.4, 127.5, 127.6, 127.8, 128.2, 128.3, 128.4, 137.8,

138.5, 138.9, 141.1, 145.8, 146.3, 163.7; MALDI-TOF-MS: 9911.02 ([M+H]+, calcd. for

C618H659N48O72: 9911.12).

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N N3

OBnBnO

BnO

BnO12

3

45 7

6

8

9

10

11

12

4

N N3

OBnBnO

BnO

BnO12

3

45 7

6

8

9

10

11

12

4

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N N3

OHHO

HO

HO12

3

45 7

6

8

9

10

11

12

5

N N3

OHHO

HO

HO12

3

45 7

6

8

9

10

11

12

5

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N N

OHHO

HO

HO12

3

45 7

6

8

9

10

11

12

6

NN

13

1416 17

15

N N

OHHO

HO

HO12

3

45 7

6

8

9

10

11

12

6

NN

13

1416 17

15

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Figure S1. 1H (top) and 13C NMR (bottom) spectra of compound 8 recorded in DMSO-d6.

The 1H NMR spectrum of 8 shows the typical signal of the 1,2,3-triazole unit at δ 7.81 ppm.

The 13C NMR spectrum of fullerene hexakis-adduct 8 is in full agreement with its T-

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symmetrical structure and shows the expected signals for the 6 equivalent malonate addends.

Only 3 signals out of the 5 expected ones are however observed for the fullerene C atoms (δ =

69.0 for the sp3 C atom; 140.7 and 144.9 ppm for the sp2 C atoms). Indeed, these 3 signals

are reminiscent of those of the three non-equivalent fullerene C atoms of the hexakis-adduct

carrying achiral addends (overall Th symmetry). No influence of the overall symmetry of 8

which is T could be deduced and the two pairs of diastereotopic sp2 C atoms are pseudo-

equivalent. Similar observations have been reported for related C60 derivatives.vi

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Figure S2. 1H (top) and 13C NMR (bottom) spectra of compound 9 recorded in CDCl3.

S16

Figure S3. MALDI-TOF-MS of compound 9. Mass spectra of 8 and 9 were recorded under

different conditions (MALDI-TOF, ESMS and FAB). However, in the case of 8, high level of

fragmentation prevented the observation of the expected molecular ion peak. Similar

observations have been reported for fullerene-sugar conjugates.vi In the case of protected

derivative 9, the level of fragmentation is less dramatic and the molecular ion peak could be

clearly observed at m/z 9911.02 ([M+H]+, calcd. for C618H659N48O72: 9911.12). Typical

fragments resulting from the loss of one or two malonate addends can be observed (m/z

8380.62 and 6849.74). Other fragments result from the cleavage of ester functions followed or

not by decarboxylation.

S17

Figure S4. UV/vis spectra of 8 (recorded in H2O, top) and 9 (recorded in CH2Cl2, bottom).

The UV/vis spectra of compounds 8 and 9 show the characteristic features of fullerene hexa-

adducts.vi,vii

400 6000,0

0,2

0,4

0,6

λ (nm)

OD

400 6000,0

0,1

0,2

0,3

0,4

0,5

0,6

OD

λ (nm)

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General Procedures for Inhibition Assay. The glycosidases β-glucosidase (from bovine

liver, cytosolic), α-galactosidase (from Aspergillus niger), α-galactosidase (from green coffee

beans), β-glucosidase (from almonds), amyloglucosidase (from Aspergillus niger), α-

glucosidase (from yeast), isomaltase (from yeast), naringinase (Penicillium decumbes), β-

mannosidase (from Helix pomatia) and α-mannosidase (from jack bean) used in the inhibition

studies, as well as the corresponding o- and p-nitrophenyl glycoside substrates, were

purchased from Sigma Chemical Co. Inhibitory potencies were determined by

spectrophotometrically measuring the residual hydrolytic activities of the glycosidases against

the respective o- (for β-glucosidase/β-galactosidase from bovine liver) or p-nitrophenyl α- or

β-D-glycopyranoside, in the presence of the corresponding iminosugar derivative. Each assay

was performed in phosphate or phosphate-citrate (for α- or β-mannosidase or

amyloglucosidase) buffer at the optimal pH for each enzyme. The Km values for the different

glycosidases used in the tests and the corresponding working pHs are listed herein: β-

glucosidase (bovine liver), Km = 2.0 mM (pH 7.3); α-glucosidase (yeast), Km = 0.35 mM (pH

6.8); β-glucosidase (almonds), Km = 3.5 mM (pH 7.3); α-galactosidase (coffee beans), Km =

2.0 mM (pH 6.8); amyloglucosidase (Aspergillus niger), Km = 3.0 mM (pH 5.5); naringinase

(Penicillium decumbes), Km = 2.7 mM (pH 6.8); β-mannosidase (Helix pomatia), Km = 0.6

mM (pH 5.5); α-mannosidase (jack bean), Km = 2.0 mM (pH 5.5). The reactions were

initiated by addition of enzyme to a solution of the substrate in the absence or presence of

various concentrations of inhibitor. After the mixture was incubated for 10-30 min at 37 ºC

the reaction was quenched by addition of 1 M Na2CO3. The absorbance of the resulting

mixture was determined at 405 nm or 505 nm. The Ki value and enzyme inhibition mode were

determined from the slope of Lineweaver-Burk plots and double reciprocal analysis using a

Microsoft Office Excel 2003 program. Data represent mean standard deviation (n = 3).

Representative plots are reproduced hereinafter.

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Figure S5. Lineweaver-Burk Plot for Ki determination (0.71±0.09 µM) of 6 against

amyloglucosidase (Aspergillus Niger) (pH 5.5).

-5

0

5

10

15

20

-1,0 0,0 1,0 2,0 3,0 4,0

1/V

1/[S] (mM-1)

Ι = 0 µΜ

Ι = 0.25 µΜ

Ι = 0.5 µΜ

Ι = 1 µΜ

Ι = 2 µΜ

Ι = 4 µΜ

S20

Figure S6. Lineweaver-Burk Plot for Ki determination (0.69±0.06 µM) of 8 against

amyloglucosidase (Aspergillus Niger) (pH 5.5).

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Figure S7. Lineweaver-Burk Plot for Ki determination (10.5±0.9 µM) of 8 against isomaltase (baker

yeast) (pH 6.8).

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Figure S8. Lineweaver-Burk Plot for Ki determination (0.41±0.04 µM) of 8 against naringinase

(Penicillium decumbes) (pH 6.8).

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Figure S9. Lineweaver-Burk Plot for Ki determination (0.15±0.02 µM) of 8 against Jack beans α-

mannosidase (pH 5.5).

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i M.-Y. Chen, J.-L. Hsu, J.-J. Shie, J.-M. Fang, J. Chin. Chem. Soc. 2003, 50, 129-133. ii H. S. Overkleeft, J. van Wiltenburg, U. K. Pandit, Tetrahedron 1994, 50, 4215-4224. iii B. Jagadish, R. Sankaranarayanan, L. Xu, R. Richards, J. Vagner, V. J. Hruby, R. J. Gillies, E. A. Mash, Bioorg. Med. Chem. Lett. 2007, 17, 3310-3313. iv A. J. Rawlings, H. Lomas, A. W. Pilling, J.-R. L. Lee, D. S. Alonzi, J. S. S. Rountree, S. F. Fenkinson, G. W. J. Fleet, R. A. Dwek, J. H. Jones, T. D. Butters, Chem. Bio. Chem. 2009, 10, 1101-1105. v Compound 7 was prepared as described in: J. Iehl, J.-F. Nierengarten, Chem. Eur. J. 2009, 15, 7306-7309. vi J.-F. Nierengarten, J. Iehl, V. Oerthel, M. Holler, B. M. Illescas, A. Muñoz, N. Martín, J. Rojo, M. Sánchez-Navarro, S. Cecioni, S. Vidal, K. Buffet, M. Durka, S. P. Vincent, Chem. Commun. 2010, DOI:10.1039/C0CC00034E. vii A. Hirsch, O. Vostrowsky, Eur. J. Org. Chem. 2001, 829-848.