Supporting Information

 Wiley-VCH 2010
69451 Weinheim, Germany

Glycosidase Inhibition with Fullerene Iminosugar Balls: A Dramatic Multivalent Effect**

Philippe Compain,* Camille Decroocq, Julien Iehl, Michel Holler, Damien Hazelard, Teresa Mena Barragn, Carmen Ortiz Mellet,* and Jean-Franois Nierengarten*

anie_201002802_sm_miscellaneous_information.pdf

Supplementary Information

S1

Table of Contents

General Methods Syntheses and Analytical data of the Compounds

1

S3 S3 S10 S16 S17 S18 S24

H and 13C NMR Spectra of the Compounds

MALDI-TOF-MS of compound 9 UV/vis spectra of 8 and 9 General Procedures for Inhibition Assay References

S2

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 BIFLEX
TM

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 (20C) in a PerkinElmer 241 polarimeter for sodium ( = 589 nm).

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

OBn BnO BnO 1 O

NH3 (30%), I2

OBn BnO BnO OH NH 2

THF, r.t. overnight OBn OH

O 2 OBn

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 S3

white solid. The analytical data of 2 were in complete agreement with those reported in the literaturei: 1H NMR (300 MHz, CDCl3): = 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--lactam (3)

OBn BnO BnO OH NH2

1) DMSO, Ac2O BnO BnO 2) NaCNBH3, HCOOH 3 CH 3CN

OBn NH O OBn

O 2 OBn

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 0C 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 0C 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): = 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).

S4

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

OBn BnO BnO 3 NH O OBn

LAH THF, reflux

OBn BnO BnO NH 10 OBn

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 0C. The reaction mixture was heated under reflux for 2 h, then cooled at 0C. 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): = 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)

OBn BnO BnO NH 10 OBn

Br NEt3, DMAP, DMF 120 C,4 days

N3 BnO BnO

OBn N 4 OBn

N3
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 120C 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 S5

5:1) gave 4 (57 mg, 38%) as a yellow oil. [ ]D -5 (c 1, CHCl3). 1H NMR (300 MHz, CDCl3):
20

= 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, H7B), 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);
13

C NMR (75 MHz, CDCl3): = 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)

OBn BnO BnO 4 N OBn

N3
5

BCl3 HO HO 5

OH N OH

N3
5

CH 2Cl2 -60C 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 60C. The resulting mixture was allowed to warm to 0C 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. [ ]D -13 (c 1, MeOH). 1H NMR (300 MHz, CD3OD): = 1.29-1.52 (m, 4H),
20

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);
13

C NMR (75 MHz, CD3OD): = 25.0, 27.6, 27.9, 29.8, 52.4, 53.7, 57.1, 58.6,

S6

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)

H OH HO HO 5 N OH N3
5

CuSO4 Sodium Ascorbate H 2O/DMF 1:1

OH HO HO 6 N OH

N N N
5

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 50C 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. [ ]D -10 (c 0.82, MeOH). 1H NMR (300 MHz,
20

CD3OD): = 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): = 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).

S7

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): = 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): = 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.

S8

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): = 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): = 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).

S9

BnO BnO
3 2

OBn
7 8 9 10 11 12

N
1

N3

BnO

BnO BnO
3 2

OBn
7 8 9 10 11 12

N
1

N3

BnO

S10

HO HO
3 2

OH
7 8 9 10 11 12

N
1

N3

HO

HO HO
3 2

OH
7 8 9 10 11 12

N
1

N3

HO

S11

HO HO
3 2

OH
7 8 9 10 11 12

N
1

N N 14 N
13

15 16 17

HO

HO HO
3 2

OH
7 8 9 10 11 12

N
1

N N 14 N
13

15 16 17

HO

S12

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 7.81 ppm. The
13

C NMR spectrum of fullerene hexakis-adduct 8 is in full agreement with its TS13

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 ( = 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 pseudoequivalent. Similar observations have been reported for related C60 derivatives.vi

S14

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

S15

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.

S16

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 hexaadducts.vi,vii

0,6

0,4

OD

0,2

0,0 400

(nm)

600

0,6

0,5

0,4

OD
0,3

0,2

0,1

0,0 400

(nm)

600

S17

General Procedures for Inhibition Assay. The glycosidases -glucosidase (from bovine liver, cytosolic), -galactosidase (from Aspergillus niger), -galactosidase (from green coffee beans), -glucosidase (from almonds), amyloglucosidase (from Aspergillus niger), glucosidase (from yeast), isomaltase (from yeast), naringinase (Penicillium decumbes), mannosidase (from Helix pomatia) and -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 -glucosidase/-galactosidase from bovine liver) or p-nitrophenyl - or -D-glycopyranoside, in the presence of the corresponding iminosugar derivative. Each assay was performed in phosphate or phosphate-citrate (for - or -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: glucosidase (bovine liver), Km = 2.0 mM (pH 7.3); -glucosidase (yeast), Km = 0.35 mM (pH 6.8); -glucosidase (almonds), Km = 3.5 mM (pH 7.3); -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); -mannosidase (Helix pomatia), Km = 0.6 mM (pH 5.5); -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 C 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.

S18

Figure S5. Lineweaver-Burk Plot for Ki determination (0.710.09 M) of 6 against

amyloglucosidase (Aspergillus Niger) (pH 5.5).

20
= 0 = 0.25 = 0.5

15

1/V 10
5

= 1 = 2 = 4

0 -1,0 -5 0,0 1,0 2,0 3,0 4,0

1/[S] (mM-1)

S19

Figure S6. Lineweaver-Burk Plot for Ki determination (0.690.06 M) of 8 against

amyloglucosidase (Aspergillus Niger) (pH 5.5).

S20

Figure S7. Lineweaver-Burk Plot for Ki determination (10.50.9 M) of 8 against isomaltase (baker
yeast) (pH 6.8).

S21

Figure S8. Lineweaver-Burk Plot for Ki determination (0.410.04 M) of 8 against naringinase

(Penicillium decumbes) (pH 6.8).

S22

Figure S9. Lineweaver-Burk Plot for Ki determination (0.150.02 M) of 8 against Jack beans mannosidase (pH 5.5).

S23

M.-Y. Chen, J.-L. Hsu, J.-J. Shie, J.-M. Fang, J. Chin. Chem. Soc. 2003, 50, 129-133. 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. Muoz, N. Martn, J. Rojo, M. Snchez-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.
ii

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