Carbohydrate Research 341 (2006) 1685–1691

Note

Synthesis of thioswainsonine as a potential glycosidase inhibitor
Nag S. Kumar and B. Mario Pinto*
Department of Chemistry, Simon Fraser University, Burnaby, British Columbia, Canada V5A 1S6
Received 18 March 2006; received in revised form 10 April 2006; accepted 13 April 2006 Available online 6 May 2006

Abstract—The synthesis of a bicyclic sulfonium-ion analogue of a naturally occurring glycosidase inhibitor, swainsonine, in which the bridgehead nitrogen atom is replaced by a sulfonium ion, has been achieved by a multi-step synthesis starting from 1,4-anhydro2,3-di-O-benzyl-4-thio-D -lyxitol. The synthetic strategy relies on the intramolecular displacement of a leaving group on a pendant acyclic chain by a cyclic thioether. This bicyclic sulfonium salt will serve as a candidate to test the hypothesis that a sulfonium salt carrying a permanent positive charge would be an effective glycosidase inhibitor. Ó 2006 Elsevier Ltd. All rights reserved.
Keywords: Glycosidase inhibitor; Swainsonine analogue; Bicyclic sulfonium salt; Indolizidine alkaloid analogue

Glycosidases play an important role in the metabolism of carbohydrates and in the processing of specific oligosaccharide structures on glycoproteins; the latter play important roles in intercellular recognition processes, and their modification has been implicated in disease states such as cancer.1–7 Tumor cells display very complex carbohydrate structures that are usually restricted to embryonic tissues and it is believed that these structures provide signal stimuli for rapid proliferation and metastasis of tumor cells. Because tumor and normal cells have different rates of cell growth, a glycosidase inhibitor can be used to block the assembly of complex oligosaccharide structures, which might lead, in turn, to a therapeutic strategy for the treatment of cancer. For example, swainsonine (1), a plant-derived alkaloid, is an excellent inhibitor of Golgi a-mannosidase II (GMII) and has been shown to reduce tumor cell metastasis, enhance cellular immune responses, and slow tumor growth in mice.8 Treatment with swainsonine (1) has led to a significant reduction of tumor mass in humans suffering from breast, liver, lung cancer, and other malignancies.9,10 The most important class of reversible glycosidase inhibitors is composed of naturally occurring polyhydroxylated pyrrolidine, piperidine, pyrrolizidine, and indolizidine alkaloids11–17 which
* Corresponding author. Tel.: +1 604 291 4152; fax: +1 604 291 4860; e-mail: bpinto@sfu.ca 0008-6215/$ - see front matter Ó 2006 Elsevier Ltd. All rights reserved. doi:10.1016/j.carres.2006.04.009

are monocyclic and bicyclic amines such as 1-deoxynojirimycin (2), castanospermine (3), and swainsonine (1). It is postulated that these amines bind to glycosidase enzymes by mimicry of the shape and charge of the oxacarbenium ion transition state for the enzyme-catalyzed hydrolysis reaction.18,19 The nitrogen atom is known to be protonated in the enzyme active site, thus providing the stabilizing electrostatic interactions between the inhibitor and the carboxylate residues in the enzyme active site. An alternative means of achieving the required charged state to provide such electrostatic interactions would be to synthesize compounds that incorporate a permanent positive charge at a suitable position.

N OH

OH

HO HO

H N OH OH 2

OH 1

HO HO

N OH

OH 3

which was subsequently reduced using sodium borohydride to furnish compound 17. After cyclization.20 the synthesis. Pinto / Carbohydrate Research 341 (2006) 1685–1691 O BnO BnO 12 7 steps Ref. B. as shown in Scheme 2. Compound 18 was treated with sil- . Cl S OH OH HO OH 4 OH 5 HO ClO4 S D-Lyxose 3 steps Ref. The 1.22 and the synthesis of sulfonium ions (6. The secondary hydroxyl group at C-5 in each of compounds 15a and 15c was protected as a benzyl ether to give compounds 16a and 16c. S.3. 30 S HO BnO 11 OBn S BnO BnO 13 OBn OBn OH Recently. OTf S HO 6 OH OH HO 7 S OH OTf OH Cl S S OH 8 OH O OH 9 S I S O Cl OH OH OH 10 Retrosynthetic analysis indicates that the bicyclic sulfonium salt A could be synthesized by an intramolecular displacement of a suitable side-chain leaving group by a cyclic thioether (Scheme 1).24–26 Most recently they have reported the synthesis of compound 8. in turn. Treatment of 17 with methanesulfonyl chloride furnished the corresponding sulfonate ester. we described the synthesis of the sulfoniumion analogue of 3-epi-swainsonine (4).5-tri-O-benzyl-4-thio-D -lyxitol (13).4-anhydro2..26 The syntheses of di-O-methylated sulfonium derivatives (9)27 of swainsonine (1). which is not only a potent inhibitor of several mannosidases. which were separated by flash chromatography. facilitated by the aldehyde group in 14. free and enzyme-bound conformations. along with a trace amount of diastereomer 15c (Scheme 3).30 The primary benzyl ether was selectively removed using 1% sulfuric acid in acetic anhydride to give the corresponding acet8 7 6 5 L S 4 1 2 3 L 8 OP 7 6 5 S 4 1 2 3 OP 5 S 4 1 2 3 OP OP A OP OP B OP OH OP C P = protecting group L = leaving group Scheme 1. as well as 5-O-methylthioswainsonine28 have also been reported in the literature.23 Siriwardena and co-workers have also reported the synthesis of bicyclic sulfonium salts to serve as glycosidase inhibitors. was synthesized from commercially available D -lyxose. which was subsequently treated with a freshly prepared Grignard reagent from the reaction of 2-(2-bromoethyl)-1. 7) of the pyrrolizidine alkaloid. but shows greater selectivity than swainsonine (1).3-dioxane protecting group in compound 16a was hydrolyzed to give the corresponding aldehyde. Most of the minor isomer 15b was recrystallized out from the mixture using ethyl acetate and hexane.5-Tri-O-benzyl-D -lyxofuranoside (12) was prepared in three steps starting from D -lyxose. Compound 11. ate which was subsequently removed by methanolysis to give compound 11. transformed using a multi-step procedure into 1.29 which was. respectively (Scheme 4). Swern oxidation of compound 11 using trifluoroacetic anhydride furnished aldehyde 14. The key intermediate B could. be synthesized from C. corresponding to C.1686 N. Kumar.3. it was found that the major compound 15a had the required R configuration at C-5.21. M. Compound 15c was the result of epimerization at C-4. and glycosidase inhibitory activity (albeit weak) of a sulfonium-ion analogue (5) of castanospermine (3). as described by Postema et al. which was subsequently treated with LiBr to give bromide 18. 2. in turn.3-dioxane and magnesium to give inseparable diastereomers 15a and 15b as a 2:1 mixture. australine. We now report the first synthesis of the sulfonium-ion analogue (10) of swainsonine (1) as a potential glycosidase inhibitor. 29 1) 1% H2SO4/Ac2O 2) NaOMe/MeOH 65% (2 steps) Scheme 2.

ii) LiBr / THF / Reflux 90% 2 steps OH OBn S BnO 17 OBn i) BCl3 / CH2Cl2 / -78oC ii) Ion Exchange (Amberlyst A-26) 70% Cl S OH 10 OH OBn 19 OH Scheme 4.31 The configuration at C-5 of compound 19 was assigned by means of a 1D-NOESY experiment. Experimental 1.20 1.1. the stereochemistry of compound 15a was thus assigned by inference. A mixing time of 500 ms was used in the . OBn 15a / 15c NaH / DMF BnBr O O BnO OBn 16c (7%) S O O OBn S BnO OBn 16a (88%) i) AcOH / H2SO4/ H2O / 1. The benzyl protecting groups were removed with boron trichloride at À78 °C and the product was subsequently treated with Amberlyst A-26 (chloride form) to completely exchange the triflate counterion with chloride ion to give compound 10. 1H and 13C NMR spectra were recorded with frequencies of 500 and 125 MHz. 1D-NOESY experiments were recorded at 295 K on a 500 MHz spectrometer. ver triflate in acetonitrile to promote cyclization. The ring junction of this bicyclic compound 19 was cis as expected. B. R2 = OH (minor) OH O O BnO OBn 15c (trace) S 1687 S O DMSO / TFAA 11 DIPEA / CH2Cl2 H BnO Scheme 3.N.13C (gHMQC) experiments using standard Varian pulse programs. M. 512 scans were acquired with a Q3 Gaussian Cascade pulse. For each 1D-NOESY spectrum. Kumar.4-dioxane (78:0. which showed a correlation between H-3 and H-5. S. because our previous work had shown that some of the triflate counterion was exchanged to chloride ion during the course of deprotection. Pinto / Carbohydrate Research 341 (2006) 1685–1691 R1 R2 O O S OBn 14 O THF 68% (2 steps) dr. giving the desired sulfonium salt 19 as a stable.5) ii) NaBH4 / EtOH 71% (2 steps) OBn S Br AgOTf / CH3CN 88% OTf S OBn OBn BnO 18 OBn i) MsCl / Pyr. General methods Optical rotations were measured at 23 °C and reported in deg dmÀ1 gÀ1 cm3.1H (gCOSY) and 1H. Processing of the spectra was performed with MestRec software. respectively.28. R2 = H (major) 15b: R1 = H. 2:1(15a:15b) MgBr O BnO OBn 15a: R1 = OH.5:20:1.20. colorless oil. All assignments were confirmed with the aid of two-dimensional 1H.

55 g.b and 15c: MALDI-TOF MS: m/z 445. 3. 21. 4. J1a. Column chromatography was performed with silica gel 60 (230–400 mesh). 1H.28 (m. H-4). 483.97 [M+K]+.72 (ddd. H-3). 10.2 (2Cipso).3 0 -dioxan-2 0 -yl)L -gluco-heptitol (15b) and 1.3-di-O-benzyl-4-thio-7(1 0 . 51.29 (m.06.55 (ddd. filtered. 4.91 (ddd. 128.5b = 4.54 (dd. 81. 368.15 (m. and the mixture was stirred at À78 °C for 30 min.64 (m.84 [M+Na]+.75 mL. 1.11 (C-6 0 ). 11.0 (C-1).1688 N.5 (C-4).77 (dd. A Grignard reagent. .06 mmol) in CH2Cl2 (15 mL) was added dropwise while maintaining the temperature below À78 °C and the stirring was continued for 1. 3. 2H. 2. filtered.6 (C-4). H-5). 1.5 (C-5).7a = J2 0 .79 and 4.6 0 eq = J5 0 ax. filtered.08 (dd.1 Hz. CH2Ph).8 (10CAr). Kumar.2 = 4.54.15– 4. hexanes/EtOAc) to give the aldehyde 14.1 Hz. J4 0 ax. 10H. hexanes/ EtOAc) to give a 2:1 mixture of compounds 15a and 15b. J5a.7 Hz. 138. B.3 = 3. H. along with a trace amount of compound 15c as a white solid (0.7 (10CAr).4 (C-7).3. 1.79–1.5% molybdic acid in 10% aqueous H2SO4.5 M.9 mmol) in 1.b = 11. H7a. Pinto / Carbohydrate Research 341 (2006) 1685–1691 1D-NOESY experiments. H-1a).4 = 5. The reaction mixture was stirred at ambient temperature for 14 h and then the reaction was quenched by the addition of 0.2 = 4.01 g. Ja. 72.2 = 6. 67.07 (ddddd.0 Hz.7-dideoxy-2. The reaction mixture was stirred at room temperature for 1 h and then neutralized with HOAc. J2. and concentrated.7 (C-6). 13C NMR (CDCl3): d 138. 102.7 (C-2).4 = 3. The organic phase was washed with H2O (100 mL) and brine (100 mL). 1H.5 0 ax = J5 0 ax.2 (C-3).64 (2d. Anal.2 Hz. 67.30 [M+K]+.6 mmol) in CH2Cl2 (5 mL) dropwise. 1H.98 and 4. H-5 0 ax). 73. The crude product was purified by flash chromatography (6:1.5 mmol) in CH2Cl2 (15 mL) at À78 °C under N2 atmosphere was added a solution of trifluoroacetic anhydride (0. 1H. Data for the major diastereomer (15a): 1H NMR (CDCl3): d 7.5 M HCl (5 mL).3 0 -dioxan-2 0 -yl)-D -manno-heptitol (15a). Calcd for C25H32O5S: C.49 mmol) and Mg (272 mg.38–7. 13C NMR (CDCl3): d 138. The residue was diluted with Et2O (200 mL) and washed with H2O (2 · 50 mL) and brine (50 mL). Compound 14 was diluted with THF (15 mL) and cooled to 0 °C under N2.6 Hz.61 (2d. H-7b). and heated.b = 11.6 Hz.21 [M+H]+.4.8 Hz. H-2 0 ).90 mmol) in THF (15 mL). followed by brine (100 mL).9 Hz. 1. 3. 1. MALDI-TOFMS: m/z 331.4-anhydro-6.0% H2SO4/Ac2O (50. 3H. Matrix-assisted laser desorption ionization time-of-flight (MALDI-TOF) mass spectra were obtained using 2. 5.5 h. 128. 3. The organic layer was washed with H2O (100 mL). J3.7b = 4. The organic layer was dried over anhydrous Na2SO4. 68%). M.5 0 eq = 12. 1 H NMR (CDCl3): d 7.5a = 7.6 0 ax = 2.2 = 9. each 1H.5 Hz.4 Hz. 79. The organic phase was dried over anhydrous Na2SO4. The organic layer was dried over anhydrous Na2SO4. 1H.3-di-O-benzyl-4-thio-D -lyxitol (11) A solution of 1. H. Ja. J4. 1H.9 Hz. 1H.5 Hz.92 (dd. The reaction was quenched by the addition of aqueous HCl (0.1. 63. A solution of diisopropylethylamine (2.58 (2d.7. 47.31 (m. J1a. The crude product was purified by flash chromatography (3:1. 138. H-4). 4.9 (c 1.02 (ddd.45.71.7-dideoxy-2. J3. The Et2O layer was washed with H2O (50 mL) and brine (50 mL).9.2 (C-1). 3.2. Found: C. Ja. 12. 5 mL) and the mixture was partitioned between Et2O (200 mL) and H2O (50 mL). 81. A solution of compound 11 (1.43–1. H-4 0 eq.1b = 9. 67.2 Hz.3-dioxane (0.6 mmol) in CH2Cl2 (15 mL) was added dropwise and the stirring was continued at À78 °C for an additional 2 h. 7.6 (C-3).b = 12. For the mixture of 15a. 3.20. H-1b).1 Hz. 31.8–127. Ja. 1H. The minor isomer (15b) was obtained by crystallization from hexanes/EtOAc as a white solid: mp 116–118 °C.b = 12. J4. 1. 2. Calcd for C19H22O3S: C. 7.3-di-O-benzyl-4-thio-7-(1 0 . 72.3 Hz. 71. 1H. 6.7-dideoxy-2.87 mL. H-6 0 eq).4 (C-2 0 ). Found: C.74 (2d. 1H.72 (ddd. H-6a. H-5b). each 1H. H-6b).4-Anhydro-2. J1a. H-4 0 ax). satd aq NaHCO3 (2 · 100 mL). H-5a).63 and 4. CH2Ph). 31. J1a. 83.27 (dd.4 0 eq = J4 0 ax5 0 ax = 12. J4 0 eq.4 Hz. and concentrated under reduced pressure. 1H. 6. J5 0 eq. J1b. 1.5b = 11.3 0 -dioxan-2 0 -yl)-(D -talo/L -allo)heptitol (15c) To a stirred solution of DMSO (1. 10H.5-tri-O-benzyl-4-thio-D lyxitol (13) (5. respectively. 467.9 Hz. 352.3 mL.9 (C-5 0 ). 4. H-5 0 eq. 2. 25. The mixture was concentrated and then partitioned between EtOAc (300 mL) and H2O (100 mL). 3. 4. 4.4 (2CH2Ph).5 mL.0 Hz.92 (dd.40–7.0 mL) was stirred at ambient temperature for 14 h and then partitioned between EtOAc (200 mL) and H2O (100 mL).6 0 ax = 12. 2H. H-1a).0 (2Cipso).6a = 9. J2 0 . 11.92 g.1 (C-5). J4. 67.5 = J5. 4. H-2.9 Hz. H-2). H.85. and concentrated. S.19 (dd. 29. H. The developed plates were air-dried. The crude product was purified by flash chromatography (3:1. 69. 1H.17–4. was added dropwise.13 (C-4 0 ). Analytical thin-layer chromatography (TLC) was performed on aluminum plates precoated with silica gel 60F-254 as the adsorbent. Anal.2 Hz.30 [M+H]+. 4.4-anhydro-2.4-anhydro6. H-6 0 ax).20.6b = 2.3di-O-benzyl-4-thio-7-(1 0 .69 and 4. and concentrated. H-1b).1 Hz. each 1H. CH2Cl2). 3.4-Anhydro-6. The crude product was dissolved in CH3OH (200 mL) and 1 M NaOCH3/CH3OH solution was added until the solution was basic. J6 0 ax. 2H.4 (2CH2Ph).32 [M+Na]+.6 0 eq = 4. 73. 3. Ar). 69. The organic phase was dried over anhydrous Na2SO4.1 Hz. CH2Ph). each 1H.5 0 eq = 2. J5.08 (m.5 (C-2). 31.04 (dd.90 (dd. J1b.0 Hz.1. 65%): [a]D À12. filtered. hexanes/EtOAc) to give compound 11 as a colorless oil (2. exposed to UV light and/or sprayed with a solution containing 1% Ce(SO4)2 and 1. 1H.7–127. J4 0 ax. CH2Ph). 1H. freshly prepared from 2-(2-bromoethyl)-1. H-3).4 Hz. 3. Ar). J5 0 ax.5-dihydroxybenzoic acid as a matrix.27.3. 4.1b = 10.00 g. 1H. and concentrated.

84.6b = 14. 4. 1H.N. 25.50 (dd.2 Hz.50 (dd.5 0 ax = 12. H-5 0 eq).2 Hz. 137.32 (2d. J50 ax.5 Hz. J1a.6 0 eq = J5 0 ax. Anal. Pinto / Carbohydrate Research 341 (2006) 1685–1691 1689 [a]D +5. 30.7a = 10. 1. 1. 1. H.27 [M+K]+. J4 0 eq.2 (3Cipso). 3. 467.7 Hz.0 Hz. 15H. 4. C-4 0 . 67. 1H.1.3 0 dioxan-2 0 -yl)-(D -talo/L -allo)-heptitol (16c) A mixture of compounds 15a/15c (145 mg.3. Data for the minor diastereomer (16c): [a]D À17. Anal.1 (C-5). J4 0 eq.5 = 7.27.54.4.68 (ddd. Ja. 7.7 (C-7).3 = J3. J7a.2 Hz.24 (m. CH2Cl2). H-6a). J6a.8 Hz. J5 0 eq.7 (C-1).6a = 4.2 = 10. 52. H-3. MALDI-TOFMS: m/z 535.5-tri-O-benzyl-4-thio-7-(1 0 .7-dideoxy-2. Ja. 1H. 1. 4. Calcd for C32H38O5S: C.2 Hz.99.4. = J7b. CH2Ph). H-2 0 ). 3. 81. H-6a.4 Hz.2 = 8. 3. The residue was diluted with Et2O (50 mL) and washed with H2O (20 mL) and brine (20 mL).16.25 mmol) in 5 mL AcOH– H2O–H2SO4–1. H-4 0 eq. 0. J5 0 ax. 4. each 1H. 13C NMR (CDCl3): d 138. 82. J1b.59 (2d. 1H. 2. each 1H. Ar).2 Hz. MALDI-TOFMS: m/z 445.0 Hz.09–4.5 0 ax = J5 0 ax.3 equiv) was added and the solution was stirred at room temperature for 2 h. 30.3 Hz. 1H.46 and 4. Data for the major diastereomer (16a): [a]D À5.5 0 eq = J5 0 eq. 1. 7.5 0 eq = 2. 3. J6a.36–7. J1a. 3.0 (C-5 0 ).5 0 eq = 2.1 (c 0. 1H. CH2Ph). 30. The crude product was purified by flash chromatography (3:1. Ja.42 (2d.26 [M+H]+. J1a.7a = 9.0 Hz.7-dideoxy-2.7 Hz. 1H. 1H. 1H. 3.07 and 4. 1. J6a. J6b.1 (C-2 0 ).8. 1H. CH2Ph).00 (dd.7b = 11. 138. 13C NMR (CDCl3): d 139. H-1b). H-7a). H-4 0 ax).2 (3CH2Ph).6 Hz.4 0 eq = J4 0 ax. 1H.95 (dd.5 = 9. J6a.b = 11.5 0 eq = 12.51 (dddd. J7a. 0. H-5 0 ax). 102.0 Hz. 4. H-5 0 ax).5 Hz.5 equiv) in DMF (8 mL) was stirred in an ice bath for 15 min. 3. J4 0 ax. H-1a).0 (C-5 0 ).98 (ddd. 10H. H-2).5-tri-O-benzyl-4-thio(17) D -manno-octitol A solution of 16a (135 mg.3 Hz. 7.71 (ddd.3 (C-2 0 ).31 (m. 1H. 2. J6 0 ax. CH2Ph).9 (10CAr).4 (C-7). Benzyl bromide (66 lL. 71.9 (2C. H-4 0 ax).7 (2Cipso). J2.7– 127. 7. 102.4-Anhydro-6.3.3.27.13–4. C-6 0 ).88. H-2.1 Hz.29 (m. 3.46 (m. H-7b). 1H. H4 0 eq.04 (m.4 Hz.96. 1H. 1H. J1a. J4 0 ax.3 = J3. 79.6 (C-5). J6a.6 (C-3). 73. 2H. J4.5:1. 15H.16–4.10 (C-4 0 ).7a = 5.31 (m.6 0 eq = 1. J40 eq. J1a.1 Hz. H-3). 573.05. 1. The organic phase was dried over anhydrous Na2SO4.5 0 eq = 13. H. filtered.65 (dddd.3 (c 1.4 Hz. H-4).6 Hz. 3. The residue was diluted with 95% EtOH (30 mL) and the solution was cooled to 0 °C.01 (ddd.6 0 ax = 2.27. 71. 1H. 71%): mp 98– 100 °C.08 [M+H]+. J5 0 ax.7b = 14. C-6 0 ).8 (C-5 0 ).9. H-2 0 ).16. H-4 0 eq.b = 12. each 1H. 1. 1H. The reaction was quenched by the addition of AcOH (0.0 Hz. and concentrated.69 (ddd.2 (c 0.4 = 4. and concentrated.49.86 and 4. 483.32. J6b.9 (C-1). 1H.7b = 5. 1H.6 (C-3). H-7b).b = 11. 2H. J5 0 eq.1 Hz.95 [M+K]+. J6a. 7. 30.2 0 = J7b. 71. 3. M. 7. H-1b).59 (2d.6 0 eq = J5 0 ax.04 (ddddd. 78. H-5 0 eq). 80. MALDI-TOFMS: m/z 535. 51. 138.4 = 2. 3.33 mmol) and 60% NaH (20 mg.2 0 = 5. 67.7 (C-2). H-5). 138. 1.4 (C-4). 25. H. H-6a).70 and 4. Calcd for C25H32O5S: C. 4. H-7b). 1H NMR (CDCl3): d 7. J6 0 ax.7b = 8. 1H NMR (CDCl3): d 7. 1H. Anal.9 (C-1). J1b. H-7a. H-6b. H-1a). 66.2 = 5.43 (dd. hexanes/ EtOAc) to give 16a (153 mg. 572. hexanes/EtOAc) to give compound 17 as a white solid (86 mg.5 Hz.6.5.7 Hz.5 (C-6). 1H.7a = J2 0 .9 Hz.63 (2d. 2. Kumar. 73. 13C NMR (CDCl3): d 138. satd aq NaHCO3 (2 · 20 mL).50 eq = 12.6 0 eq = 4. Ja.6 0 ax = 12. Ja.2 0 = 3.8–127.60 eq = 5. 88%) and 16c (12 mg.4-dioxane (78:20:0.b = 11. CH2Cl2). 72. 128. J4.5 Hz.0 Hz H-4).0 (c 0. 1H.35 (m.0 Hz.80. and brine (20 mL).93 (dddd.74 (ddd. H-1a). S.b = 11. H-5 0 eq).4 Hz.94 (dd.77 (dddd.13 (dd.4 Hz. 5.5 = J5. J2.4-Anhydro-6. The reaction was quenched by the addition of ice H2O (1 mL) and the mixture was diluted with Et2O (50 mL). Ja. Ar). H. J4 0 ax. 71.59. 4. H-7a. The mixture was diluted with Et2O (50 mL) and washed with H2O (20 mL).5 (15CAr). 3.56 (s. 1.6 (15CAr).4 0 eq = J4 0 ax.77–1. 1H.6b = 14.96 (ddd. 31. Found: C.08 (C-6 0 ).7a = 4.0 Hz.5 0 ax = J5 0 ax. 49.7 (C-2). CH2Ph). [a]D À6.4.72 (ddd.15 [M+H]+. The crude product was purified by flash chromatography (5:1.36–7.05 (dd.4 (C-4). H-6 0 ax). J3.53 (dddd.6 Hz. 1H.46 (2d. J4 0 ax. 102.6 0 ax = 2. CH2Cl2).1b = 10.56 and 4.6 0 ax = 11. CH2Cl2).7 Hz. 67. H-1b). H.1b = 10. H-6 0 ax).6 Hz. H-6 0 eq). Found: C. 4. 1H.5) was stirred at room temperature for 20 h.34 (dd.2 = 6. 4.1 Hz. 77.37–7. 2. J6b.6 Hz. 1H.b = 12.35.44 (dd. 1H. each 1H.6 0 eq = J5 0 ax.6 0 ax = 2. J5 0 ax. J5.5-tri-O-benzyl-4-thio-7(1 0 .4 Hz. 26.5 (2CH2Ph). 2.4 (C-7). 1.12 [M+Na]+. 2H.9 Hz. The organic layer was washed with H2O (20 mL) and brine (20 mL). 1H.06 (ddddd. 1H.8 Hz. 1H.02 (ddd.5. J4 0 eq.4anhydro-6. 1. 1H. H-3). 26.71 (ddd.88. 1 equiv) was added and the mixture was stirred in an ice bath for 1 h.36 (3CH2Ph). H-6b) 1.4 Hz. 4H.8–127.6b = 8. NaBH4 (10 mg.2. H-4).7b = 5. J4. 1H.6b = 4.0 Hz. each 1H. H-6 0 ax). 71.59 (m. 71. 1H NMR (CDCl3): d 7. Ar).2 = 5. J1a. 4.3 Hz.6a = J5.7b = 6. 3.30 [M+Na]+. 1. 25. H-5 0 ax).0 Hz. J6b.03 (m. 1H. The organic phase was dried over anhydrous Na2SO4.7 (C-5). CH2Ph). 67.5 Hz. 1.31 (ddddd. 556. each 1H.78–1.96 [M+Na]+.8 (C-3). H-6 0 eq).4 = 3. J1b. CH2Ph). 4. CH2Ph). Found: C.0 Hz. 3.7 Hz.11 (3Cipso).5 (C-2 0 ). 4. 7%). 72. 1H.6 0 eq = 5.85 (dd.1 (C-2). 71. J7a. H-6b). 4H.04 (ddddd.42 (dd.62 and 4.6 Hz.2 = 7.b = 11. C-4 0 . 30.8 Hz.5 0 eq = 2.3. J4 0 ax. H-6 0 eq). Ja.05 (m.7 Hz. J2 0 .5 0 ax = 12. 128.58 (2d. 2H.3 (C-4).2 mL) and the mixture was concentrated. 67. 557. 4.6 0 ax = 11.5 0 eq = 12. 1H. H-4 0 ax).63 (m.55 and 4.03 (dd.5.32 [M+K]+. Calcd for C32H38O5S: C.1 (2C.22 (m. 79.7-dideoxy-2. filtered.3 0 -dioxan-2 0 -yl)-D -manno-heptitol (16a) and 1.4 0 eq = J4 0 ax. 1. 69. 72. and concentrated.4 Hz. 4. 1. 4. The organic phase was dried over anhydrous Na2SO4.5 Hz. H-2). H-2 0 ). 4. each 1H.9 Hz H-5).1 Hz.4 (C-6). 3. 1H. 1H NMR (CDCl3): . J4 0 ax. 1H. 29. 2.5 Hz. B.5 0 ax = 12. J6 0 ax.1b = 9.50 ax = J50 ax. 4. J5 0 eq. H.9 Hz.1 (C-6).1. H-5). 128. 138. filtered.60 (dd. J5.

1C.2 (c 0. 4.37 (2d. 1H.4 Hz. H-7ax.5-Trihydroxy-cis-1-thioniabicyclo[4.6b = 3. and concentrated.34 (dd. H-7eq). H-8a). CH3OH). H-2). 74.01.00. 1H. Anal.98 (ddd. 3. 1H.1 Hz. J1a.2 = 6. 64. J1a.56 (ddd.7–127. J7ax.5 = 10. H-8b).29 [MÀBr]+.14 (ddddd. CH2Cl2–CH3OH) to give compound 19 as a colorless syrup (55 mg. hexanes/EtOAc) to give compound 18 as a colorless oil (61 mg. H-7b). Ja.b = 11. each 1H.7–127.4-Anhydro-8-bromo-6. H-4). CH2Ph). 7. H.9 Hz. JC.36–7.6b = 3.2 equiv). MALDI-TOF MS: m/z 478. 25. Ja. each 1H.9 Hz. 3.5 Hz. 2. 27.39 (br s. The residue in THF (4 mL) together with LiBr (55 mg. 1H. 1. CH2Ph).4 = 2.45.76 (m. H. (ddd. 129. Calcd for C29H34O4S: C. H-4).20 (m.4 = 3. J1a. MALDI-TOFMS: m/z 461. each 1H. 3. H-1a). 1H NMR (CD3OD): d 4. Anal.2 = 7.3. J1a.b = 11.2 = 9.98 and 4.4 (C-8). Another equivalent of AgOTf (26 mg) was added and the reaction mixture was stirred for an additional 10 h. J1b.17 (dd. Ja.2 (c 0. 3. J7a.7eq = 3.8 (C-4).34 (ddd. 4. Pinto / Carbohydrate Research 341 (2006) 1685–1691 d 7.9 Hz.2 Hz. CH2Ph).0 Hz. H-7b). 1H. H-3). and concentrated. 43. 138.36 (dddd. 13C NMR (CDCl3): d 139. 5. 5 equiv) was heated at reflux under N2 for 2 h and then concentrated. 3. 516.6 Hz.3 = J3.5(R)-2.8-trideoxy-2. 78. CH2Ph).9 Hz. H-6eq.3 = 2. 3.0 Hz. 1H.0]-nonane chloride (10) BCl3 gas was bubbled vigorously through a solution of 19 (50 mg. 1H NMR (CDCl3): d 7.4(R).52. 79. 78.61 (2d.7 Hz. J7ax. 2(S).6 (15CAr). 1H.1 Hz. 2H.6eq = J6ax. H. 0.2 Hz.1 (3Cipso). J4. 4.12 and 4.5 Hz. J8ax. J5.42 (m. H-7a.6 (C-3).35 (dd.3 (C-7). 15H. H-3).8 (c 0. 1.8a = 5.2 (C-7). J1a. 1H. 3. Calcd for C29H33BrO3S: C. 49. H-5).58. 7. 5. J4. 59.95 (dd.23 (m.7 Hz. 72. 4. 49.9 (C-2). 27.2 Hz.1b = 12. 1.0 Hz. OTf).3.3 Hz. 15H.2 (C-4).79 (br s.15 (dd.3 Hz. 69. each 1H.1.2 = 10. 1H. H-3. H-2. CH2Cl2).1690 N. 4. 2(S). J7a. 63. 72.8a = J7b.52–4. 2. CH2Ph).85 (ddd. 1H. 1H. J1a.8eq = 13. CH2Ph). each 1H. H. 3. 13C NMR (CDCl3): d 139. The organic phase was washed with H2O (20 mL).6 (C-3). H. J2. each 1H.2. J8ax. Ar). 1H. 136. filtered. 2. Ar). 30.08. The organic phase was dried over anhydrous Na2SO4. 29. H-4). 3.8ax = 11. 3. 4.5.3 = J3. J1b. Ja.6ax = 2. S.8. 82 lmol) in CH2Cl2 (5 mL) at À78 °C under N2 for 10 min. 1H. J1a. 82.59 and 4. 2. 59.68 (m. 1H. 1.7 (C-1). 1H. 1H.91 [M+K]+. J5.7ax = 14.19 (dd. 4H. 1 equiv) for 14 h at ambient temperature.00 (ddd.2 = 10.6. 128. [a]D À6. Ja. H-6b.4(R).2 Hz.13 mmol) in pyridine (4 mL) at 0 °C under N2 was added methanesulfonyl chloride (12 lL. J7eq. H-8ax). 1H. H-2). The filtrate was concentrated and recrystallized from CH3OH–CH2Cl2 to give compound 10 as white crystals (13 mg. H-6a. H-1b). MALDI-TOFMS: m/z 461. H-8eq).1b = 12. J3.50 (m.4 (C-4). 3. 34.97 (dd. 5.4 Hz. 3.46 (2d.3 Hz. 1H.36 (m.1 (3CH2Ph).10 mmol) in CH3CN (3 mL) was treated with AgOTf (26 mg.58 (2d. H-8ax).07 [M+Na]+. Found: C. 3H. 3.1 (3Cipso). J7ax. J8a.8 (C-6). The solid was dissolved in CH3OH (5 mL) and a freshly washed ion exchange resin (Amberlyst A-26 (chloride form). 128. 4. 3.8ax = 10.5 (C-5).b = 11.16. 3H. 4. 64.01–1.9 (C-2).37– 7. The mixture was stirred at À78 °C for 2 h and a stream of dry air was blown vigorously over the solution to remove excess BCl3. J1b.43 (dd.7 Hz. B. H-1a).66 and 4.3. J6ax.6 Hz. Ar). H-3). H-5). 5.6a = 9.8ax = 3. 4.59 (2d.5-tri-Obenzyl-4-thio-D -manno-octitol (18) To a stirred solution of 17 (60 mg.b = 12. 75. 138. 72.9 (q.7 Hz. J2.4 Hz. 4. 1H NMR (CDCl3): d 7. H-1b). H-2). 15H. 60 mg) was added. Ja. 84. H.32.7 (C-5). H-8eq).0 Hz.).3. 4. 0.5 Hz. CH2Cl2). The organic layer was dried over anhydrous Na2SO4.53 and 4. 1H.5 Hz.1. 1H. filtered. 77. 3. Anal. 1 M HCl (20 mL). H-6ax. Kumar. 84.54 (dd.3. Calcd for C30H33F3O6S2: C.06 (ddd.5(R)-2.5 = J5. 4.14.6. CH2Ph). H-1b).85–1. The mixture was stirred at room temperature for 30 min and filtered.3 (C-8).6 (15CAr). The solvent was removed and the crude product was purified by flash chromatography (20:1. 1H.0]-nonane triflate (19) A solution of compound 18 (55 mg.2 Hz.37 (2d.1. 1H. 88%): [a]D À38.5.b = 11. J1b.3 Hz. 73. each 1H.25 [MÀOTf]+. Ja.56 (m. 4H. 78.8 Hz. H-1a). 4. 2. 1H.0 Hz.2 (15CAr).65 (m. 40.32. 4. 1H.1 Hz.65 (2d.80 (dd.b = 11.9 Hz. Ja. J7eq. J7b. 27.b = 12.7. 30.62 (ddd. 3.5 (C-1). 138. 1. each 1H.6 (C-6).27.5. H-7a.15 (dd.50 (br s. 1H. Found: C.8.4 (C-3). 1H.5-Tribenzyloxy-cis-1-thioniabicyclo[4.9 Hz. 1.8eq = 14. M. 13 C NMR (CDCl3): d 137.8ax = 2. 52.0 Hz.61 and 4. 1. H-5).1 Hz.6b = 4. The residue was diluted with Et2O (50 mL) and washed with H2O (2 · 20 mL) and brine (20 mL). 137. CH2Ph). 501.56 (2d. 2.41–3. 71. H-8a.2 = 7. H-6a. H-1a).5.7. The mixture was concentrated under high vacuum and then diluted with Et2O (50 mL).7 (C-1).3(R).4 = 3.8.51 and 4. 6. 70%): mp > 219 °C (decomp.6 Hz.3(R). 4. 1H.58 (2d. 6.05–1. 71.8.11 and 4.3 (C-7).2 = 6.b = 11. H-6b).8 (C-2).8eq = J6eq.0–128. 16.8 (C-6).77. H-5).3. 1H.6 (3CH2Ph).0 (C-5). Found: C.8a = 7.1b = 9.38–7. H-4). satd aq NaHCO3 (20 mL).8 Hz. each 1H. The reaction was quenched by the addition of CH3OH (2 mL) and the solvent was removed. The reaction mixture was stirred at 0 °C for 2 h and then quenched by the addition of ice (0. 3. J2.8 (3CH2Ph). 71. The residue was co-evaporated with CH3OH (2 · 5 mL) and then washed with CH2Cl2 (2 · 2 mL) to give a white solid.F = 318.21 (m. H-6ax). 138. H-1b).9 Hz. CH2Ph). J5. H-8b). J5. 4.69 and 4.2 = 10. 73. and brine (20 mL). The crude product was purified by flash chromatography (5:1.8b = 5.4 Hz. 90%): [a]D À6. J6ax. 1H.1b = 10.2.b = 11.89 (m.5 mL). 73.13 (dd.93 (br s.8 (3Cipso).1.0 Hz. Ja. 120.3 (C-8).7ax = .97 [M+H]+. 1H.24.05 (ddd. J1a.8b = 10.

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