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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).
S5
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
S6
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).
S8
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.
S9
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).
S10
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
S11
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
S12
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
S13
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-
S14
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
S15
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)
S18
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.
S19
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).
S21
Figure S7. Lineweaver-Burk Plot for Ki determination (10.5±0.9 µM) of 8 against isomaltase (baker
yeast) (pH 6.8).
S22
Figure S8. Lineweaver-Burk Plot for Ki determination (0.41±0.04 µM) of 8 against naringinase
(Penicillium decumbes) (pH 6.8).
S23
Figure S9. Lineweaver-Burk Plot for Ki determination (0.15±0.02 µM) of 8 against Jack beans α-
mannosidase (pH 5.5).
S24
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.