Syntheses of glycoclusters containing a phosphocholine residue related to a glycosphingolipid from the earthworm Pheretima hilgendorfi

Article abstract of DOI:10.1248/cpb.57.1081

Three types of glycoclusters related to an amphoteric glycosphingolipid found in the earthworm Pheretima hilgendorfi were synthesized. The glycoclusters were prepared from a common precursor and a simple approach for the rational design of a glycocluster was developed.

Full text of DOI:10.1248/cpb.57.1081

October 2009
Chem. Pharm. Bull. 57(10) 1081—1088 (2009)
1081
Syntheses of Glycoclusters Containing a Phosphocholine Residue Related
to a Glycosphingolipid from the Earthworm Pheretima hilgendorfi
Noriyasu HADA,*,a Yukihiko SHIDA,a Natsuko NEGISHI,a Frank SCHWEIZER,b and Tadahiro TAKEDA*,a
a Faculty of Pharmacy, Keio University; 1–5–30 Shibakoen, Minato-ku, Tokyo 105–8512, Japan: and b Department of
Chemistry, University of Manitoba; Winnipeg, Manitoba, R3T 2N2, Canada.
Received May 8, 2009; accepted July 21, 2009; published online July 24, 2009
Three types of glycoclusters related to an amphoteric glycosphingolipid found in the earthworm Pheretima
hilgendorfi were synthesized. The glycoclusters were prepared from a common precursor and a simple approach
for the rational design of a glycocluster was developed.
Key words glycocluster; amphoteric glycosphingolipid; Pheretima hilgendorfi; phosphocholine
In our continuing studies to investigate the relationship be- Results and Discussion
tween the structure and biological function of glycolipids
Synthesis of Disaccharide Units 6 and 9 In order to
from invertebrate animal species that do not have ganglio- synthesize the glycoclusters access to the disaccharide units
sides, we have synthesized glycolipids found in various pro- 6 and 9 was required (Chart 1). Monosaccharide derivative 3
tostomia phyla.1—12) These compounds may serve as ganglio- was obtained by condensation of phenyl 2,3,4-tri-O-benzoyl-
side mimics. Sugita et al. reported13) the neogala series of 6-O-tert-butyldiphenylsilyl-1-thio-b-D-galactopyranoside (1),
glycosphingolipids, whose structures contain a b-D-Galp- prepared by silylation and benzoylation of phenylthio-b-D-
(1→6)-b-D-Galp-core and phosphocholine residue, found in galactopyranoside,20) with the spacer 2 in the presence of N-
the earthworm Pheretima (P. ) hilgendorfi, and we previously iodosuccinimide (NIS) and trifluoromethanesulfonic acid
synthesized two phosphocholine (PC) glycolipid analogues (TfOH) in CH2Cl2 in 79% yield.21,22) The anomeric proton of
containing octyl residues in place of ceramide, PC(→6)-b-D- the galactose unit appeared as a doublet at d 4.72 (d, Jꢀ7.9
Galp-1→Oct and PC(→6)-b-D-Galp-(1→6)-b-D-Galp-1→ Hz). Selective removal of the tert-butyldiphenylsilyl (TBDPS)
Oct to investigate the biological function of zwitterionic group in 3 with tetrabutylammonium fluoride (TBAF) gave
oligosaccharides.6) In later studies it was found that the disaccharide acceptor 4, which was subjected to glycosyla-
disaccharide PC(→6)-b-D-Galp-1→Oct has immunomodu- tion with thioglycosyl donor 1 in the presence of NIS/TfOH
latory functions leading to induced production of interleukin to afford the desired disaccharide 5 in 89% yield. The b-gly-
(IL)-12 and tumor necrosis factor (TNF)a by macrophages cosidic linkage was assigned on the basis of homonuclear
and dentritic cells and others.14) In a previous paper,12) we re- coupling constants (H-1ꢁ, dꢀ4.80 ppm, JH1ꢁ,H2ꢁꢀ7.9 Hz). Se-
ported on the total synthesis of two phosphocholine (PC) gly- lective removal of the Troc-protecting group from 5 by
cosphingolipids, PC(→6)-b-D-Galp-(1→6)-b-D-Galp-1→Cer Zn–AcOH gave the primary amine 6. On the other hand, re-
and PC(→6)-b-D-Galp-(1→6)-b-D-Galp-(1→6)-b-D-Galp- moval of the TBDPS group was achieved by treatment of 5
1→Cer and other related analogues, in order to investigate with TBAF to give 7. Phosphorylation with phosphoryl chlo-
another immunomodulatory functions of zwitterionic oligo- ride followed by exposure of the resulting dichloroester to
saccharides. In addition, we also examined the potential of choline tosylate yielded choline derivative 8. Deblocking of
these newly synthesized glycosphingolipids and analogues to the Troc group from 8 with Zn–AcOH produced the primary
enhance production of IL-8 in TNFa-stimulated granulocytic amine 9.
HL-60 cells. Our results demonstrated that the phospho-
choline and ceramide groups are potent enhancers of IL-8 were studied for the synthesis of glycocluster A (Chart 2).
production in TNFa-stimulated granulocytic HL-60 cells. Initially, we attempted to install the PC group at the end of
Synthesis of Phenyl-Core Glycocluster A Two methods
It is known that oligosaccharide chains generally interact the synthesis after cluster formation. Trimesoyl chloride was
with their protein receptors in a multivalent fashion to over- selected as the core for the synthesis of the phenyl-core clus-
come the inherently low affinity of monovalent carbohy- ter. N-Acylation of trimesoyl chloride with amine 6 in the
drate–protein interactions. Therefore, the construction of a presence of triethylamine afforded the trivalent oligosaccha-
clustered glycoconjugates is an important subject in glyco- ride 10 in 56% yield. Removal of the TBDPS group using
science.15,16) For this reason, we developed a new synthetic TBAF afforded alcohol 11 in 73% yield. Phosphorylation of
method of glycoclusters containing a sugar unit and w-amino alcohol 11 was carried out by a multi-step procedure using a
acid.17) Here we report on the synthesis of three types of gly- phosphorodiamidite method.23,24) Initially, 11 reacted with 2-
coclusters A, B and C (Fig. 1). Phenyl core cluster A was cyanoethyl N,N,Nꢁ,Nꢁ-tetraisopropylphosphorodiamidite fol-
chosen as a model cluster due to the high reactivity and lowed by exposure to choline tosylate. The obtained crude
rigidity of the trifunctional trimesic acid scaffold.18) Glyco- product was oxidized in situ by m-chloroperbenzoic acid
dendrons B and C were selected based on increased flexibil- (mCPBA) and the cyanoethyl protecting group was removed
ity and variation in the number of attached amphoteric disac- using aqueous ammonia in methanol. Chromatographic pu-
charide units. 4-Aminobutanoic acid (GABA) was used as a rification of the final product afforded 6-O-phosphocholine
flexible linker unit due to expected proteolytic stability19) and disaccharide 12 in a low yield (9%). To improve the yield of
commercial availability.
12 we also studied phosphorylation of 11 with phosphoryl
© 2009 Pharmaceutical Society of Japan
∗ To whom correspondence should be addressed. e-mail: hada-nr@pha.keio.ac.jp

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Vol. 57, No. 10
Fig. 1. Target Compounds
Reagents: (a) NIS, TfOH, CH2Cl2, MS 4 Å, 3: 79%; 5: 89%; (b) TBAF, AcOH, THF; 4: 87%; 7: 81%; (c) Zn, AcOH; 6: quant.; 9: quant.; (d) (i) POCl3, Et3N, MS 3 Å,
CH2Cl2, (ii) choline tosylate, pyr., (iii) H2O; 69% (three steps).
Chart 1
chloride and the resulting dichloroester was immediately amino diacid 20 as a new core unit for the preparation of gly-
converted to the phosphocholine derivative 12 using choline codendrons B and C. Compound 20 required access to two
tosylate in 58% yield.25) In the second method we studied building blocks 3-(benzyloxycarbonyl)-1-propanol (14) and
cluster formation after incorporation of the PC-group. This 4-(4-nitro-benzenesulfonylamino)-butanoic acid benzyl ester
was achieved by coupling of phosphocholine containing di- (16). Benzyl ester 14 was prepared by basic hydrolysis of
saccharide 9 with trimesoyl chloride to afford 12 in 71% 4-butyrolactone using aq. NaOH followed by esterfication
yield. Finally, removal of the benzoyl groups in 12 under using benzyl bromide and tetrabutylammonium bromide in
Zemplén conditions, followed by column chromatography acetone in 85% yield. The second building block 16 was pre-
(Sephadex LH-20), furnished the target phenyl-core glyco- pared from GABA in a two step procedure. At first, GABA
cluster A (13).
was converted into benzylester under acidic conditions fol-
Synthesis of Glycodendrons B and C We selected lowed by protection of the amino group p-nitrobenzenesul-

October 2009
1083
fonyl (pNBS) chloride to give 16. The two building blocks
Next, we examined the outcome of the condensation of the
16 with 14 were coupled using Mitsunobu conditions26) to core unit 20 with the PC-containing disaccharide unit 9.
produce 17. Deblocking of the pNBS group with PhSH and However, coupling of 20 with 9 in the presence of diethyl-
K2CO3 in N,N-dimethylformamide (DMF)27) followed by car- cyanophosphate (DEPC) and Et3N was not successful (data
bamoylation with di-tert-butyl dicarbonate ((Boc)2O) af- not shown). Fortunately, coupling of 20 with sugar unit 6 in
forded benzyl ester 19. Finally, basic hydrolysis of ester 19 the presence DEPC and Et3N was successful to produce de-
gave dicarboxylic acid derivative 20 (Chart 3).
sired divalent derivative 21 in 73% yield. The tert-butoxy-
carbonyl (Boc) group of 21 was removed under acidic condi-
tions using 50% CF3CO2H in CH2Cl2 and the generated sec-
ondary amino group was acylated using 2-(tetradecyl)hexa-
decanoic acid and N-[3-(dimethylamino)propyl]-Nꢁ-ethylcar-
bodiimide hydrochloride (EDC) to give 23 in 87% yield. De-
blocking of the tert-butyldiphenylsilyl (TBDPS) group was
achieved by treatment of 23 with TBAF to produce 24 in
52% yield. Phosphorylation of 24 was peformed with phos-
phoryl chloride, and the resulting dichloroester was immedi-
ately converted to the phosphocholine derivative 25 by expo-
sure to choline tosylate. Finally, de-O-benzoylation of 25 af-
forded symmetric glycodendron 26 (B) (Chart 4). Similarly,
glycodendron 32 (C) was prepared by coupling of building
blocks 20 with 22 to produce the desired tetramer derivative
27 in quantitative yield. Deblocking was performed as de-
scribed above for dimer dendron 26 affording unprotected
glycodendron 32 (C) (Chart 5). All of the glycoclusters de-
scribed in this paper were purified by column chromatogra-
Reagents: (a) trimesoyl chloride, Et3N, CH2Cl2, 10: 56%, 12: 71%; (b) TBAF,
AcOH, THF, 73%; (c) (i) 2-cyanoethyl-N,N,Nꢁ,Nꢁ-tetraisopropylphosphorodiami-
dide, 1H-tetrazole, CH2Cl2, MS 3 Å, (ii) 1H-tetrazole, choline tosylate, (iii)
mCPBA, MeOH, (iv) aq. NH3; 9% (four steps); (d) (i) POCl3, Et3N, MS 3 Å,
CH2Cl2, (ii) choline tosylate, pyr., (iii) H2O, 58% (three steps); (e) NaOMe,
MeOH, 79%.
Reagents: (a) (i) NaOH, H2O, (ii) BnBr, Bu4NBr, acetone, 85%; (b) BnOH,
TsOH, toluene, 98%; (c) pNsCl, Et3N, CH2Cl2, 88%; (d) Ph3P, DEAD, CH2Cl2,
78%; (e) PhSH, K2CO3, MeCN, 93%; (f) (Boc)2O, Et3N, CH2Cl2, 61%; (g) NaOH,
H2O, dioxane, quant.
Chart 2
Chart 3
Reagents: (a) DEPC, Et3N, DMF, 73%; (b) TFA, CH2Cl2, 75%; (c) 2-(tetradecyl) hexadecanoic acid, EDC, DMAP, CH2Cl2, 87%; (d) TBAF, AcOH, THF, 52%; (e) (i)
POCl3, Et3N, MS 3 Å, CH2Cl2, (ii) choline tosylate, pyr., (iii) H2O, 54% (three steps); (f) NaOMe, MeOH. 84%.
Chart 4

1084
Vol. 57, No. 10
Reagents: (a) DEPC, Et3N, DMF, 99%; (b) TFA, CH2Cl2, 94%; (c) 2-(tetradecyl) hexadecanoic acid, EDC, DMAP, CH2Cl2, 90%; (d) TBAF, AcOH, THF, 85%; (e) (i)
POCl3, Et3N, MS 3 Å, CH2Cl2, (ii) choline tosylate, pyr., (iii) H2O, 21% (three steps); (f) NaOMe, MeOH, 80%.
Chart 5
phy on silica gel or Sephadex (LH 20). The solubilities of
all final compounds (A, B, C) containing phosphocholine
were excellent in methanol and in a mixture of 1 : 1
methanol–water. The purified final compounds were charac-
J
1,2ꢀ7.9 Hz, J2,3ꢀ10.4 Hz, H-2), 5.62 (1H, dd , H-3), 4.81 (1H, br s, NH),
4.72 (1H, d, H-1), 4.72—4.68 (2H, m, COOCH2CCl3), 4.06 (1H, t,
5,6ꢀ7.3 Hz, H-5), 3.93—3.89 (1H, m, CH2CH2O), 3.83 (2H, d, H-6),
J
3.50—3.46 (1H, m, CH2CH2O), 3.04—3.00 (2H, m, CH2NH), 1.55—1.04
(8H, m, CH2ꢃ4), 1.00 (9H, s, t-Bu); 13C-NMR (125 MHz, CDCl3) d: 165.6,
165.4, 165.2, 154.4, 135.5, 135.4, 133.2, 133.1, 132.5, 130.0, 129.8, 129.73,
29.66, 129.6, 129.5, 129.0, 128.5, 128.3, 128.2, 127.7, 127.6, 101.6, 74.4,
73.7, 71.9, 70.1, 70.0, 67.9, 61.3, 41.0, 29.4, 29.1, 26.6, 26.1, 25.4, 19.0;
MALDI-TOF-MS: Calcd for C52H56Cll3NNaO11Si: m/z 1026. Found: 1026.8
[MꢂNa]ꢂ.
1
terized by commonly available techniques, such as H- and
13C-NMR spectroscopy and time of flight mass spectrometry.
Conclusions
In summary, an efficient synthetic procedure for glycoclus-
ters 13 (A), 26 (B) and 32 (C) containing a phosphocholine
residue related to glycosphingolipids from the earthworm
Pheretima hilgendorfi has been developed. It is expected that
these glycocluster will induce enhanced immune responses
when compared to their monovalent counterparts.
6-N-(2,2,2-Trichloroethoxycarbonyl)aminohexyl 2,3,4-Tri-O-benzoy-
b-D-galactopyranoside (4) A solution of 3 (1.0 g, 0.99 mmol) and acetic
acid (0.2 ml, 3.0 mmol) in THF (2.5 ml) was treated with 1 M TBAF in THF
(2.0 ml, 2.0 mmol) at room temperature and then was stirred for 12 h. After
concentration, the residue was added to the water, extracted with CHCl3, and
the organic layer was proceeded as usual. The product was purified by silica
gel column chromatography (toluene : acetoneꢀ4 : 1) as eluent to give 4
(667 mg, 87%). MALDI-TOF-MS: Calcd for C36H38Cll3NNaO11: m/z 788.
Found: 788.9 [MꢂNa]ꢂ.
Experimental
General Methods Optical rotations were measured with a Jasco P-1020
digital polarimeter. 1H- and 13C-NMR spectra were recorded with a JMN
A500 FT NMR spectrometer with Me4Si as the internal standard for solu-
tions in CDCl3 CD3OD. Matrix assisted laser desorption/ionization-time of
flight (MALDI-TOF)-MS was recorded on a Perseptive Voyager RP mass
spectrometer. High-resolution mass spectra were recorded on a JEOL JMS-
700 under FAB conditions. TLC was performed on Silica Gel 60 F254 (E.
Merck) with detection by quenching of UV fluorescence and by charring
with 10% H2SO4. Column chromatography was carried out on Silica Gel 60
(E. Merck).
6-N-(2,2,2-Trichloroethoxycarbonyl)aminohexyl 2,3,4-Tri-O-benzoyl-
6-O-(tert-butyldiphenylsilyl)-b-D-galactopyranosyl-(1→6)-2,3,4-tri-O-
benzoy-b-D-galactopyranoside (5) A solution of compound 1 (2.2 g,
2.6 mmol) and 4 (1.7 g, 2.2 mmol) containing activated MS 4 Å (1.7 g) in dry
CH2Cl2 (9.0 ml) was stirred under an atmosphere of argon for 2 h at room
temperature. After cooling to 0 °C, successively NIS (0.74 g, 3.3 mmol) and
TfOH (39.0 ml, 0.44 mmol) were added and stirring was continued at 0 °C
for 30 min, then neutralized with Et3N. The reaction mixture was filtered,
and the filtrate was washed with aqueous sodium thiosulfate, dried (MgSO4),
and concentrated. The product was purified by silica gel column chromatog-
raphy (toluene : acetoneꢀ10 : 1) as the eluent to give 5 (2.9 g, 89%). [a]D24
ꢂ93.0 (cꢀ1.4, CHCl3); 1H-NMR (500 MHz, CDCl3) d: 8.05—7.04 (40H,
m, 8Ph), 6.00 (1H, d, J3ꢁ,4ꢁꢀ3.7 Hz, H-4ꢁ), 5.81 (1H, d, J3,4ꢀ3.7 Hz, H-4),
5.68—5.63 (2H, m, H-2, 2ꢁ), 5.66 (1H, dd , J2ꢁ,3ꢁꢀ10.4 Hz, H-3ꢁ), 5.48 (1H,
dd, J2,3ꢀ10.4 Hz, H-3), 4.80 (1H, d, J1ꢁ,2ꢁꢀ7.9 Hz, H-1ꢁ), 4.73—4.68 (2H,
m, COOCH2CCl3), 4.57 (1H, d, J1,2ꢀ7.9 Hz, H-1), 4.09—4.06 (2H, m,
H-5ꢁ, 6ꢁa), 3.97 (1H, t, J5,6ꢀ7.3 Hz, H-5), 3.80 (1H, dd, J5,6ꢁbꢀ5.5 Hz,
6-N-(2,2,2-Trichloroethoxycarbonyl)aminohexyl 2,3,4-Tri-O-benzoyl-
6-O-(tert-butyldiohenylsilyl)-b-D-galactopyranoside (3)
A solution of
compound 1 (2.0 g, 2.4 mmol) and 2 (1.4 g, 4.9 mmol) containing activated
MS 4 Å (2.0 g) in dry CH2Cl2 (10 ml) was stirred under an atmosphere of
argon for 2 h at room temperature. After cooling to 0 °C, successively NIS
(1.1 mg, 4.9 mmol) and TfOH (112 ml, 1.3 mmol) were added and stirring
was continued at 0 °C for 30 min, then neutralized with Et3N. The reaction
mixture was filtered, and the filtrate was washed with aqueous sodium thio-
sulfate, dried (MgSO4), and concentrated. The product was purified by silica
gel column chromatography (toluene : acetoneꢀ6 : 1) as the eluent to give 3
J6ꢁa,6ꢁbꢀ11.6 Hz, H-6ꢁb), 3.63—3.58 (3H, m, H-6a, 6b, CH2CH2O), 3.22—
3.17 (1H, m, CH2CH2O), 3.02—2.98 (2H, dd, CH2NH), 1.34—0.95 (8H, m,
CH2ꢃ4) 0.92 (9H, s, t-Bu ); 13C-NMR (125 MHz, CDCl3) d: 165.6, 165.4,
165.3, 165.1, 135.5, 133.4, 133.2, 130.0, 129.9, 129.8, 129.74, 129.69,
129.66, 129.5, 129.0, 128.5, 128.4, 128.3, 128.2, 127.7, 127.5, 101.5, 101.1,
1
(1.9 g, 79%). [a]D24 ꢂ90.3 (cꢀ1.0, CHCl3); H-NMR (500 MHz, CDCl3) d:
8.04—7.09 (15H, m, 3Ph), 6.04 (1H, d, J3,4ꢀ2.4 Hz, H-4), 5.70 (1H dd,

October 2009
1085
74.4, 73.2, 71.9, 71.6, 70.1, 69.9, 69.8, 68.6, 67.8, 67.6, 60.8, 41.0, 29.4,
29.0, 26.5, 26.1, 25.4, 18.9; MALDI-TOF-MS: Calcd for C79H78Cl3NNaO19Si:
m/z 1500.4. Found: 1500.7 [MꢂNa]ꢂ.
6-Aminohexyl 2,3,4-Tri-O-benzoyl-6-O-(tert-butyldiphenylsilyl)-b-D-
galactopyranosyl-(1→6)-2,3,4-tri-O-benzoy-b-D-galactopyranoside
To a solution of 5 (936 mg, 0.63 mmol) in acetic acid (5 ml) was added zinc
powder (2.0 g). The reaction mixture was stirred at 50 °C for 2 h. After com-
pletion of the reaction, the mixture was filtered off and washed with CHCl3.
solution of 8 (311 mg, 0.22 mmol) in acetic acid (5 ml) was added zinc pow-
der (750 mg). The reaction mixture was stirred at 50 °C for 2 h. After com-
pletion of the reaction, the mixture was filtered off and washed with CHCl3.
The filtrate was concentrated and purified by Sephadex LH-20 column chro-
(6) matography in MeOH to give 9 (285 mg, quant.). [a]D24 ꢂ38.7 (cꢀ4.5,
MeOH). 1H-NMR (500 MHz, CDCl3) d: 7.94—7.10 (30H, m, 6Ph), 5.86
(1H, d, J3,4ꢀ3.1 Hz, H-4), 5.77 (1H, d, J3ꢁ,4ꢁꢀ3.1 Hz, H-4ꢁ), 5.63 (1H, dd,
J1ꢁ,2ꢁꢀ7.9 Hz, J2ꢁ,3ꢁꢀ10.4 Hz, H-2ꢁ), 5.57—5.48 (3H, m, H-2, 3, 3ꢁ), 4.95
The filtrate was concentrated and purified by silica gel column chromatogra- (1H, d, H-1ꢁ), 4.68 (1H, d, J1,2ꢀ7.9 Hz, H-1), 4.23—4.12 (4H, m, H-5, 6ꢁa,
phy (chloroform : methanolꢀ8 : 1) to give 6 (827 mg, quant.). [a]D24 ꢂ93.0
POCH2CH2), 4.05 (1H, t, J5ꢁ,6ꢁaꢀ4.9 Hz, J5ꢁ,6ꢁbꢀ10.4 Hz, H-5ꢁ), 3.93 (1H, br,
(cꢀ1.4, CHCl3); 1H-NMR (500 MHz, CDCl3) d: 8.05—7.04 (40H, m, 8Ph), H-6a), 3.81 (1H, dd, J6ꢁa,6ꢁbꢀ9.7 Hz, H-6ꢁb), 3.59—3.49 (3H, m, POCH2-
6.00 (1H, d, J3,4ꢀ3.7 Hz, H-4), 5.81 (1H, d, J3ꢁ,4ꢁꢀ3.1 Hz, H-4ꢁ), 5.69—5.63 CH2, CH2CH2O), 3.29—3.26 (1H, m, CH2CH2O), 3.09 (9H, s, N(CH3)3),
(2H, m, H-2, 2ꢁ), 5.58 (1H, dd, H-3ꢁ), 5.48 (1H, dd, H-3), 4.80 (2H, br s,
J1ꢁ,2ꢁꢀ7.9 Hz, NH, H-1ꢁ) 4.73—4.68 (2H, m, COOCH2CCl3), 4.57 (1H, d, NMR (125 MHz, CDCl3) d: 165.3, 165.1, 165.01, 164.96, 164.93, 133.05,
1,2ꢀ7.9 Hz, H-1), 4.09—4.06 (2H, m, H-5ꢁ, 6ꢁa), 3.97 (1H, t, H-5), 3.80 132.98, 132.8, 132.7, 129.12, 129.08, 128.9, 128.8, 128.7, 128.61, 128.56,
(1H, dd, H-6ꢁb), 3.63—3.58 (3H, m, H-6, O–CH2), 3.22—3.17 (1H, m, 128.5, 128.4, 128.2, 128.1, 128.02, 127.95, 127.8, 127.64, 127.57, 100.5,
2.61 (2H, t, CH2NH), 1.31—1.07 (8H, m, CH2ꢃ4), 0.92 (9H, s, t-Bu). 13C-
J
O–CH2), 3.02—2.98 (2H, m, N–CH2), 1.33—0.92 (8H, m, alkyl); 13C-NMR
(125 MHz, CDCl3) d: 165.6, 165.4, 165.3, 165.1, 135.5, 133.9, 135.35,
133.2, 133.1, 132.6, 130.0, 129.9, 129.8, 129.74, 129.69, 129.66, 129.5,
129.0, 128.5, 128.4, 128.3, 128.2, 127.7, 127.5, 101.5 (C-1), 101.1 (C-1ꢁ),
74.4, 73.6, 73.2, 71.9, 71.6, 70.1, 69.9, 69.8, 68.6, 67.8, 67.6, 60.8, 41.0, 29.4,
29.0, 26.5, 26.1, 25.4, 18.9; MALDI-TOF-MS: Calcd for C76H77NNaO17Si:
m/z. 1327.5. Found: 1327.7 [MꢂNa]ꢂ.
100.1, 71.9, 71.6, 71.2, 69.6, 69.3, 69.0, 68.2, 67.3, 66.8, 65.6, 61.8, 58.5,
56.6, 53.2, 48.2, 38.7, 28.4, 26.7, 25.3, 24.7, 16.9; MALDI-TOF-MS: Calcd
for C65H72N2O20P: m/z 1231. Found : 1231 [MꢂH]ꢂ.
N,Nꢀ,Nꢁ-Tri-{6-[2,3,4-tri-O-benzoyl-6-O-(tert-butyldiphenylsilyl)-b-D-
galactopyranosyl-(1→6)-2,3,4-tri-O-benzoy-b-D-galactopyranoslyoxy]-
hexyl}-1,3,5-benzenetriamide (10) To a solution of 6 (830 mg, 0.63 mmol)
in CH2Cl2 (5 ml) were added triethylamine (0.12 ml, 0.85 mmol) and trime-
6-N-(2,2,2-Trichloroethoxycarbonyl)aminohexyl 2,3,4-Tri-O-benzoyl- soyl chloride (35.0 mg, 0.13 mmol). The mixture was stirred for 15 min at
b-D-galactopyranosyl-(1→6)-2,3,4-tri-O-benzoy-b-D-galactopyranoside
(7) A solution of 5 (2.4 g, 1.59 mmol) and acetic acid (0.27 ml, 4.77 mmol)
in THF (8.0 ml) was treated with 1 M TBAF in THF (3.2 ml, 3.2 mmol) at
room temperature and then was stirred for 12 h. After concentration, the
room temperature. After completion of the reaction, the mixture was con-
centrated. The product was purified by silica gel column chromatography
(toluene : acetoneꢀ6 : 1) as eluent to give 10 (306 mg, 56%). [a]D24 ꢂ71.4
(cꢀ6.9, CHCl3); 1H-NMR (500 MHz, CDCl3) d: 8.32 (3H, S, Ph), 8.27—
residue was added to the water, extracted with CHCl3, and the organic layer 7.04 (120H, m, 24Ph), 6.34 (3H, t, NH), 6.02 (3H, d, J3,4ꢀ3.7 Hz, H-4), 5.82
was proceeded as usual. The product was purified by silica gel column chro- (3H, d, J3ꢁ,4ꢁꢀ3.7 Hz, H-4ꢁ), 5.69—5.63 (6H, m, H-2, 2ꢁ), 5.59 (3H, dd,
matography (toluene : acetoneꢀ6 : 1) as eluent to give 7 (1.6 g, 81%). [a]D24
J2ꢁ,3ꢁꢀ10.4 Hz, H-3ꢁ), 5.50 (3H, dd, J2,3ꢀ10.4 Hz, H-3), 4.81 (3H, d,
1
ꢂ121.0 (cꢀ1.0, CHCl3). H-NMR (500 MHz, CDCl3) d: 8.12—7.14 (30H, J1ꢁ,2ꢁꢀ7.9 Hz, H-1ꢁ), 4.59 (3H, d, J1,2ꢀ7.9 Hz, H-1), 4.09—4.07 (6H, m, H-
m, 6Ph), 5.93 (1H, d, J3ꢁ,4ꢁꢀ3.7 Hz, H-4ꢁ), 5.83—5.78 (2H, m, H-2ꢁ, 4), 5ꢁ, 6ꢁa), 3.98 (3H, t, J5,6ꢀ7.3 Hz, H-5), 3.80 (3H, dd, J5ꢁ,6bꢁꢀ9.1 Hz,
5.69 (1H, dd, J1,2ꢀ7.1 Hz, J2,3ꢀ10.4 Hz, H-2), 5.57—5.52 (2H, m , H-3, 3ꢁ),
4.89 (1H. t, NH), 4.85 (1H, d, J1ꢁ,2ꢁꢀ7.9 Hz, H-1ꢁ), 4.73—4.68 (2H, m, (9H, m, CH2CH2O, CH2NH), 1.34—0.96 (24H, m, CH2ꢃ12), 0.92 (27H, s,
COOCH2CCl3), 4.64 (1H, d, H-1), 4.16—4.10 (2H, m, H-5ꢁ, 6ꢁa), 3.95 (1H,
t-Bu); 13C-NMR (125 MHz, CDCl3) d: 165.6, 165.4, 165.1, 135.5, 133.37,
J6ꢁa,6ꢁbꢀ11.6 Hz, H-6ꢁb), 3.63—3.58 (6H, m, H-6, CH2CH2O), 3.24—3.17
t, J5,6ꢀ6.7 Hz, H-5), 3.89 (1H, dd, J5,6ꢁbꢀ6.1 Hz, J6ꢁa,6ꢁbꢀ9.8 Hz, H-6ꢁb), 133.35, 133.1, 132.7, 132.5, 130.0, 129.9, 129.8, 129.71, 129.68, 129.6,
3.71—3.65 (2H, m, H-6a, CH2CH2O), 3.50—3.49 (1H, br, H-6b), 3.22— 129.5, 129.0, 128.5, 128.4, 128.3, 128.2, 127.7, 127.5, 101.4, 101.1, 73.6,
3.16 (1H, m, CH2CH2O), 3.02 (2H, q, CH2NH), 1.40—1.10 (8H, m, CH2ꢃ 73.1, 71.9, 71.6, 70.1, 69.9, 69.8, 68.6, 67.8, 67.6, 60.7, 40.1, 29.2, 29.0,
4). 13C-NMR (125 MHz, CDCl3) d: 166.5, 165.54, 165.46, 165.2, 165.1,
26.5, 26.4, 25.5, 18.8; MALDI-TOF-MS: Calcd for C237H231N3NaO54Si3:
154.4, 137.8, 137.7, 133.5, 133.3, 133.2, 133.1, 130.1, 129.9, 129.7, 129.4, m/z 4089. Found: 4091 [MꢂNa]ꢂ.
129.3, 129.02, 128.97, 128.8, 128.74, 128.66, 128.6, 128.5, 128.33, 128.25,
128.2, 127.9, 125.2, 101.5, 101.3, 95.7, 74.4, 73.0, 71.7, 71.6, 70.0, 69.9,
N,Nꢀ,Nꢁ-Tri-{6-[2,3,4-tri-O-benzoyl-b-D-galactopyranosyl-(1→6)-
2,3,4-tri-O-benzoy-b-D-galactopyranoslyoxy]hexyl}-1,3,5-benzenetri-
69.81, 68.77, 68.6, 68.1, 60.5, 41.0, 29.3, 29.0, 26.1, 25.4, 21.4; MALDI- amide (11) Compound 11 was prepared from 10 (256 mg) as described for
TOF-MS: Calcd for C63H60Cl3NNaO19: m/z 1262.3. Found: 1262.1 [Mꢂ preparation of 4, yielding 154 mg (73%). [a]D24 ꢂ122.3 (cꢀ0.5, CHCl3); 1H-
Na]ꢂ.
NMR (500 MHz, CDCl3) d: 8.33 (3H, s, Ph), 8.25—7.14 (90H, m, 18Ph),
6-N-(2,2,2-Trichloroethoxycarbonyl)aminohexyl 2,3,4-Tri-O-benzoyl- 6.66 (3H, t, NH), 5.91 (3H, d, J3ꢁ,4ꢁꢀ3.7 Hz, H-4ꢁ), 5.81—5.78 (6H, m, H-4,
6-O-phosphorylcoline-b-D-galactopyranosyl-(1→6)-2,3,4-tri-O-benzoy-
b-D-galactopyranoside (8) To a solution of 7 (200 mg, 0.15 mmol) and
MS 3 Å (200 mg) in dry CH2Cl2 (3.0 ml) was added phosphoryl chloride
(15.7 ml, 0.17 mmol) and triethylamine (64 ml, 0.46 mmol) at ꢄ10 °C under
2ꢁ), 5.69 (3H, dd, J1,2ꢀ7.9 Hz, J2,3ꢀ10.4 Hz, H-2), 5.58—5.51 (6H, m, H-3,
3ꢁ), 4.90 (3H, d, J1ꢁ,2ꢁꢀ7.9 Hz, H-1ꢁ), 4.64 (3H, d, H-1), 4.15—4.09 (6H, m,
H-5ꢁ, 6ꢁa), 4.00 (3H, t, J5,6ꢀ6.7 Hz, H-5), 3.92 (3H, dd, J5ꢁ,6bꢁꢀ6.7 Hz,
J6ꢁa,6ꢁbꢀ11.0 Hz, H-6ꢁa), 3.78—3.66 (6H, m, H-6b, CH2CH2O), 3.65—3.33
Ar. The solution was stirred for 1.5 h at the room temperature. To this were (6H, m, H-6a, CH2CH2O), 2.99 (3H, t, OH), 1.40—1.16 (24H, m, CH2ꢃ12);
added pyridine (3 ml) and then choline tosylate (84.3 mg, 0.31 mmol) at
0 °C. This solution was stirred for 10 h at room temperature, and were added
H2O (1 ml) and stirred for 1 h at the same temperature. After that, the solu-
13C-NMR (125 MHz, CDCl3) d: 166.54, 165.6, 165.5, 165.2, 135.1, 130.1,
129.9, 129.7, 129.4, 129.3, 129.04, 128.99, 128.8, 128.7, 128.6, 128.5,
128.4, 128.34, 128.26, 128.2, 125.3, 101.4, 101.3, 74.0, 73.2, 71.8, 71.7,
tion was filtered and concentrated. The product was purified by Iatrobeads 70.0, 69.9, 69.7, 68.8, 68.7, 68.0, 60.5, 40.1, 29.2, 29.0, 26.3, 25.4; MALDI-
column chromatography (CHCl3 : MeOH : H2Oꢀ8 : 5 : 1) as eluent to give 8
TOF-MS: Calcd for C189H177N3NaO54: m/z 3375. Found: 3377 [MꢂNa]ꢂ.
N,Nꢀ,Nꢁ-Tri-{6-[2,3,4-tri-O-benzoyl-6-O-phosphorylcoline-b-D-galac-
topyranosyl-(1→6)-2,3,4-tri-O-benzoy-b-D-galactopyranoslyoxy]hexyl}-
1
(147 mg, 69%). [a]D24 ꢂ85.4 (cꢀ3.7, CHCl3); H-NMR (500 MHz, CDCl3)
d: 8.06—7.08 (30H, m, 6Ph), 5.93 (1H, d, J3,4ꢀ2.4 Hz, H-4), 5.81 (1H, d,
J3ꢁ,4ꢁꢀ3.2 Hz, H-4ꢁ), 5.70 (1H, dd, J1ꢁ,2ꢁꢀ7.9 Hz, J2ꢁ,3ꢁꢀ9.8 Hz, H-2ꢁ), 5.66 1,3,5-benzenetriamide (12) A: method (c); To a solution of 11 (74 mg,
(1H, dd, J2,3ꢀ10.4 Hz, H-2), 5.51—5.48 (2H, m, H-3, 3), 4.99 (1H, t, NH)
31 mmol) and MS 3 Å (150 mg) in dry CH2Cl2 (2.0 ml) was added 2-cya-
4.94 (1H, d, H-1ꢁ), 4.73—4.67 (2H, m, COOCH2CCl3), 4.62 (1H, d, J1,2ꢀ noethyl-N,N,Nꢁ,Nꢁ-tetraisopropylphosphorodiamidite (61 ml, 0.19 mmol) at
7.9 Hz, H-1), 4.35 (1H, br, H-6ꢁa) 4.17—4.03 (5H, m, H-5, 6ꢁb, 6a, room temperature under Ar. The solution was stirred for 0.5 h at the room
POCH2CH2), 3.84—3.54 (5H, m, H-5, 6b, POCH2CH2, CH2CH2O), 3.28— temperature. To this were added 1H-tetrazole (27 mg, 0.39 mmol) and then
3.26 (1H, m, CH2CH2O), 3.18 (9H, s, N(CH3)3), 3.02—2.97 (2H, m, choline tosylate (142 mg, 0.52 mmol) at room temperature. This solution
CH2NH), 1.31—1.05 (8H, m, CH2ꢃ4); 13C-NMR (125 MHz, CDCl3) d: was stirred for 4 h at room temperature, and was added MeOH (2 ml) and
165.7, 165.4, 165.3, 165.1, 154.4, 143.5, 139.3, 133.4, 133.2, 133.1, 129.9,
129.8, 129.6, 129.5, 129.4, 129.2, 129.1, 129.0, 128.8, 128.6, 128.5, 128.3,
128.1, 125.8, 101.2, 100.8, 74.3, 72.8, 72.4, 71.7, 69.9, 69.6, 69.5, 68.7,
68.0, 67.6, 66.0, 61.9, 59.2, 54.1, 45.7, 40.9, 29.3, 28.9, 26.0, 25.3, 21.1,
mCPBA (27 mg, 0.16 mmol) and stirred for 1 h at the same temperature.
After that, the mixture was added 30% aq. NH3 (2 ml) and stirred for 1 h at
room temperature. The solution was filtered through a pad of Celite and con-
centrated. The product was purified by Iatrobeads column chromatography
8.48; MALDI-TOF-MS: Calcd for C68H73Cl3N2O22P: m/z 1404. Found: 1404 (CHCl3 : MeOH : H2Oꢀ8 : 5 : 1) as eluent to give 12 (10 mg, 9%). B: method
[MꢂH]ꢂ.
6-Aminohexyl 2,3,4-Tri-O-benzoyl-6-O-phosphorylcoline-b-D-galac-
topyranosyl-(1→6)-2,3,4-tri-O-benzoy-b-D-galactopyranoside (9) To a
(d); To a solution of 11 (73 mg, 30 m mol) and MS 3 Å (73 mg) in dry
CH2Cl2 (2 ml) was added phosphoryl chloride (14.9 ml, 0.10 mmol) and tri-
ethylamine (38 ml, 0.27 mmol) at ꢄ10 °C under Ar. The solution was stirred

1086
Vol. 57, No. 10
for 1 h at the room temperature. To this were added pyridine (2 ml) and then tanoic Acid Benzyl Ester (17) To a solution of 16 (3.0 g, 7.9 mmol) and
choline tosylate (50.1 mg, 0.18 mmol) at 0 °C. This solution was stirred for
10 h at room temperature, and were added H2O (1 ml) and stirred for 1 h at
14 (2.0 g, 10 mmol) in dry CH2Cl2 (30 ml) was added Ph3P (2.7 g, 10 mmol)
and 40% DEAD in toluene (4.7 ml, 10 mmol) at 0 °C, and the solution was
the same temperature. After that, the solution was filtered and concentrated. stirred for 3 h at same temperature. The mixture was washed with water,
The product was purified by Iatrobeads column chromatography (CHCl3 : dried (MgSO4), and concentrated. The product was purified by silica gel col-
MeOH : H2Oꢀ8 : 5 : 1) as eluent to give 12 (67.3 mg, 58%). C: To a solution umn chromatography using 20 : 1 toluene–acetone as eluent to give 17
of 9 (239 mg, 0.19 mmol) in CH2Cl2 (2 ml) were added triethylamine (56.8 (3.5 g, 78%). 1H-NMR (500 MHz, CDCl3) d: 8.24 and 7.91 (4H, each d, Ns),
ml, 0.40 mmol) and trimesoyl chloride (8.6 mg, 0.32 mmol). The mixture was 7.33—7.23 (10H, m, Ph), 5.10 (4H, s, benzyl methylene), 3.19 (4H, t,
stirred for 15 min at room temperature. After completion of the reaction, the
mixture was concentrated. The product was purified by Iatrobeads column CH2ꢃ2); 13C-NMR (125 MHz, CDCl3) d: 172.1, 149.6, 145.2, 135.6, 128.8,
chromatography (CHCl3 : MeOH : H2Oꢀ8 : 5 : 1) as eluent to give 12 (88.4 128.3, 128.0, 124.1, 66.1, 64.0, 47.4, 30.5, 23.4; MALDI-TOF-MS: Calcd
NH–CH2ꢃ2), 2.37 (4H, t, CH2–COꢃ2), 1.88—1.82 (4H, m, CH2–CH2–
mg, 71 %). [a]D24 ꢂ89.5 (cꢀ2.2, MeOH); 1H-NMR (500 MHz, CDCl3) d: for C28H30N2NaO8S: m/z 577. Found: 577 [MꢂNa]ꢂ.
8.63 (3H, s, Ph), 8.33—7.11 (90H, m, 18Ph), 5.95 (3H, d, J3,4ꢀ3.1 Hz, H-4),
4-(Benzyloxycarbonylpropyl-amino)-butanoic Acid Benzyl Ester (18)
5.89 (3H, d, J3ꢁ,4ꢁꢀ1.8 Hz, H-4ꢁ), 5.72 (3H, dd, J1ꢁ,2ꢁꢀ7.9 Hz, J2ꢁ,3ꢁꢀ10.4 Hz, To a solution of 17 (1.1 g, 1.9 mmol) in CH3CN (10 ml) was added K2CO3
H-2ꢁ), 5.66—5.64 (9H, m, H-2, 3, 3ꢁ), 5.06 (3H, d, H-1ꢁ), 4.81 (3H, d,
(790 mg, 5.8 mmol) and PhSH (790 ml, 7.7 mmol), and the mixture was stirred
J1,2ꢀ7.9 Hz, H-1), 4.36—4.30 (6H, m, H-5ꢁ, 6ꢁa), 4.22 (6H, br dd, for 2 h at room temperature. After concentration, the residue was diluted
POCH2CH2), 4.16 (3H, t, J5,6aꢀ4.9 Hz, J5,6bꢀ10.4 Hz, H-5), 4.01 (3H, br dd,
H-6ꢁa), 3.91—3.82 (6H, m, H-6b, 6), 3.66—3.62 (3H, m, CH2CH2O),
with CHCl3, washed with 5% HCl, aq. NaHCO3 and water, dried (MgSO4),
and concentrated. The product was purified by silica gel column chromatog-
3.59—3.62 (6H, m, POCH2CH2), 3.34—3.32 (3H, m, CH2CH2O), 3.25— raphy using 30 : 1 CHCl3–MeOH as eluent to give 18 (638 mg, 93%). 1H-
3.22 (6H, m, NHCH2), 3.18 (18H, s, N(CH3)3), 1.41—1.16 (24H, m, NMR (500 MHz, CDCl3) d: 7.36—7.29 (10H, m, 2Ph), 5.09 (4H, s, benzyl
CH2ꢃ12); 13C-NMR (125 MHz, CDCl3) d: 211.6, 168.1, 166.9, 166.8, methylene), 3.00 (4H, t, NH–CH2ꢃ2), 2.50 (4H, t, CH2–COꢃ2), 2.20—
166.7, 166.5, 149.8, 145.9, 136.5, 134.5, 134.3, 130.7, 130.4, 130.4, 2.14 (4H, m, CH2–CH2–CH2ꢃ2); 13C-NMR (125 MHz, CDCl3) d: 172.0,
130.33, 130.29, 129.91, 129.86, 129.7, 129.6, 129.5, 129.3, 129.2, 129.1, 135.6, 128.6, 128.3, 66.5, 46.6, 31.0, 21.0; MALDI-TOF-MS: Calcd for
119.5, 102.0, 101.7, 78.9, 78.7, 78.43, 73.4, 73.3, 72.9, 71.4, 73.3, 72.9,
71.4, 71.1, 70.6, 69.9, 69.1, 68.6, 67.9, 67.9, 67.3., 63.6, 60.1, 54.7, 49.5,
C22H27NNaO4: m/z 392.2. Found: 392.4 [MꢂNa]ꢂ.
4-(Benzyloxycarbonylpropyl-tert-butoxycarbonyl-amino)-butanoic
48.5, 47.7, 40.9, 30.03, 29.97, 27.4, 26.4, 9.16; MALDI-TOF-MS: Calcd Acid Benzyl Ester (19) To a solution of 18 (513 mg, 1.3 mmol) in CH2Cl2
for C204H214N6O63P3: m/z 3847. Found: 3853 [MꢂH]ꢂ.
N,Nꢀ,Nꢁ-Tri-{6-[6-O-phosphorylcoline-b-D-galactopyranosyl-(1→6)-b-
(30 ml) was added (Boc)2O (352 ml, 1.5 mmol) and Et3N (582 ml, 4.2 mmol),
and the mixture was stirred for 18 h at room temperature. The mixture was
D-galactopyranosyloxy]hexyl}-1,3,5-benzenetriamide (13; A) To a solu- diluted with CHCl3, washed with water, dried (MgSO4), and concentrated.
tion of 12 (81.1 mg, 23 mmol) in MeOH (3.0 ml) was added NaOMe (15 mg)
The product was purified by silica gel column chromatography using 20 : 1
toluene–acetone as eluent to give 19 (400 mg, 61.2%). H-NMR (500 MHz,
1
at room temperature and the mixture was stirred for 10 h, then neutralized
with Amberlite IR 120 [Hꢂ]. The mixture was filtered off and concentrated. CDCl3) d: 7.37—7.30 (10H, m, 2Ph), 5.11 (4H, s, benzyl methylene), 3.19
The product was purified by Sephadex LH-20 column chromatography in (4H, br, NH–CH2ꢃ2), 2.33 (4H, t, CH2–COꢃ2), 1.86—1.81 (4H, m, CH2–
MeOH : H2O (1 : 1) to give 13 (36.0 mg, 79%). [a]D24 ꢄ8.9 (cꢀ0.9, CH2–CH2ꢃ2), 1.43 (9H, s, t-butyl); 13C-NMR (125 MHz, CDCl3) d: 172.8,
MeOH : H2Oꢀ1 : 1); 1H-NMR (500 MHz, CD3OD) d: 8.29 (s, 3H, Ph), 4.41 155.5, 135.9, 128.5, 128.2, 79.5, 66.2, 46.2, 31.4, 28.4, 28.4, 23.8, 23.4;
(d, 3H, J1ꢁ,2ꢁꢀ7.9 Hz, H-1ꢁ), 4.32 (d, 3H, J1,2ꢀ7.9 Hz, H-1); 13C-NMR (125
MHz, CD3OD) d: 169.1, 136.4, 129.6, 104.6, 104.1, 74.83, 74.75, 74.1,
74.0, 72.0, 71.9, 69.8, 69.5, 67.2, 65.5, 60.5, 55.0, 49.9, 41.1, 30.0, 29.7,
27.3, 26.1; MALDI-TOF-MS: Calcd for C78H141N6NaO45P3: m/z 1997.
Found: 1996 [MꢂH]ꢂ.
MALDI-TOF-MS: Calcd for C27H35NNaO6: m/z 492. Found: 492 [MꢂNa]ꢂ.
4-(tert-Butoxycarbonyl-carbonylpropyl-amino)-butanoic Acid (20)
To a solution of 19 (400 mg, 0.85 mmol) in 1,4-dioxane–H2O (2 : 1, 6 ml)
was added aq NaOH (2.0 ml), and the mixture was stirred for 5 h at room
temperature. The mixture was diluted with EtOAc, washed with water, dried
(MgSO4), and concentrated to give 20 (245 mg, quant.). 1H-NMR (500 MHz,
CDCl3) d: 11.1 (2H, br s, COOH), 3.25 (4H, br, NH–CH2ꢃ2), 2.36 (4H, t,
CH2–COꢃ2), 1.88—1.83 (4H, m, CH2–CH2–CH2ꢃ2), 1.42 (9H, s, t-butyl);
13C-NMR (125 MHz, CDCl3) d: 178.6, 171.3, 80.0, 60.4, 46.1, 31.1, 28.2,
23.3, 23.1, 14.0; MALDI-TOF-MS: Calcd for C13H23NNaO6: m/z 312.
3-(Benzyloxycarbonyl)-1-propanol (14) To a solution of 4-butyrolac-
tone (5.1 g, 59.2 mmol) in H2O (59 ml) was added NaOH (2.4 g, 59.2 mmol)
at 70 °C and the mixture was stirred for 24 h. Toluene was added and evapo-
rated. To a suspension of the residue in acetone (60 ml) was added tetrabutyl-
ammoniumbromide (955 mg, 2.96 mmol), BnBr (8.46 ml, 71.1 mmol). The
reaction mixture was refluxed for 24 h, and then concentrated. The reaction Found: 312 [MꢂNa]ꢂ.
mixture was extracted with ethyl acetate. The extract was washed with 1 M
NaHSO4, NaHCO3 and water, dried (MgSO4), and concentrated. The prod-
Glycocluster 21 To a solution of core unit 20 (44 mg, 0.15 mmol) and
sugar unit 6 (500 mg, 0.38 mmol) in DMF (4.0 ml) were added triethylamine
uct was purified on silica gel column chromatography (hexane : ethyl (85 ml, 0.61 mmol) and DEPC (91 ml, 0.61 mmol). The reaction mixture was
acetateꢀ5 : 1) to give 14 (9.8 g, 85%). 1H-NMR (500MHz, CDCl3) d: stirred for 18 h at room temperature. After completion of the reaction, the
7.31—7.23 (5H, m, Ph), 5.06 (2H, s, benzyl methylene), 3.69 (1H, br, –OH) mixture was extracted with CHCl3, washed with water, dried (MgSO4), and
3.56 (2H, t, HO–CH2), 2.41 (2H, t, CH2–CO), 1.85—1.79 (2H, m, CH2–
CH2–CH2); 13C-NMR (125 MHz, CDCl3) d: 173.0, 135.5, 127.9, 127.74,
127.67, 127.5, 127.4, 65.5, 60.7, 30.1, 27.1; MALDI-TOF-MS: Calcd for
C11H15O3: m/z 195. Found: 195 [MꢂNa]ꢂ.
4-Aminobutanoic Acid Benzyl Ester (15) To a solution of 4-aminobu-
tanoic acid (5.2 g, 50 mmol), BnOH (25 ml, 0.24 mol) and TsOH (11 g, 60
mmol) in toluene (50 ml) was refluxed for 5 h. After the reaction, the residue
was crestallized from hexane and was recrestallized from ethyl acetate/
hexane to give 15 (17.9 g, 98%).
concentrated. The product was purified on silica gel column chromatography
(CHCl3 : MeOHꢀ10 : 1) to give 21 (413 mg, 73%). [a]D25 ꢂ90.1 (cꢀ4.2,
CHCl3); 1H-NMR (500 MHz, CDCl3) d: 8.06—7.03 (80H, m, 16Ph), 6.03 (2H,
d, J3ꢁ,4ꢁꢀ3.7 Hz, H-4ꢁ), 5.84 (2H, d, J3,4ꢀ3.1 Hz, H-4), 5.71—5.59 (6H, m, H-
2,2ꢁ, H-3ꢁ), 5.50 (2H, dd, H-3), 4.82 (2H, d, J1ꢁ,2ꢁꢀ7.9 Hz, H-1ꢁ), 4.59 (2H, d,
J1,2ꢀ7.9 Hz, H-1), 4.14—4.06 (4H, m, H-5ꢁ, H-6ꢁa), 3.99 (2H, t, H-5) 3.84—
3.80 (2H, m, H-6ꢁb), 3.66—3.61 (6H, m, H-6, O–CH2), 3.22 (6H, br, O–CH2,
2NCH2), 3.08—3.03 (4H, m, NHCH2), 2.12 (4H, br, NHCOCH2), 1.83—1.77
(4H, m, 2NHCOCH2CH2CH2N), 1.34—0.91 (43H, m, 3t-butyl, alkyl); 13C-
4-(4-Nitro-benzenesulfonylamino)-butanoic Acid Benzyl Ester (16) NMR (125 MHz, CDCl3) d: 165.5, 165.3, 165.23, 165.16, 165.1, 165.0,
To a solution of 15 (7.0 g, 19 mmol) in dry CH2Cl2 (70 ml) were added p-ni- 156.1, 137.7, 135.4, 135.3, 133.23, 133.15, 133.03, 132.98, 132.6, 132.4,
trobenzenesulfonyl chloride (4.7 g, 21 mmol) and Et3N (7 ml, 48 mmol), and
the mixture was stirred for 18 h at 0 °C. After the reaction, the residue was
successively washed with water, dried (MgSO4), and concentrated. The
product was purified by silica gel column chromatography using 10 : 1
toluene–acetone as eluent to give 16 (6.7 g, 88%). 1H-NMR (500 MHz,
129.9, 129.8, 129.7, 129.59, 129.56, 129.5, 129.4, 129.3, 129.0, 128.9,
128.8, 128.4, 128.3, 128.2, 128.1, 127.6, 127.4, 125.2, 101.3, 101.0, 79.6,
73.5, 73.0, 71.8, 71.6, 70.0, 69.84, 69.78, 68.5, 67.6, 67.5, 63.51, 63.46,
60.6, 53.8, 45.9, 39.3, 33.4, 29.1, 29.0, 28.9, 28.3, 26.5, 26.3, 25.4, 24.5,
23.1, 21.3, 18.7, 16.03, 15.98; MALDI-TOF-MS: Calcd for C165H173
-
CDCl3) d: 8.32 and 8.01 (4H, each d, Ns), 7.38—7.32 (5H, m, Ph), 5.24, N3NaO38Si2: m/z 2883.1. Found: 2885.0 [MꢂNa]ꢂ.
(1H, t, NH) 5.10 (2H, s, benzyl methylene), 3.07 (2H, q, NH–CH2), 2.42
(2H, t, CH2–CO), 1.86—1.81 (2H, m, CH2–CH2–CH2); 13C-NMR (125
MHz, CDCl3) d: 173.0, 150.0, 145.9, 135.5, 128.6, 128.4, 124.4, 66.7, 42.7,
Glycocluster 22 To a solution of 21 (374 mg, 0.13 mmol) in CH2Cl2
(2.0 ml) was added trifluoroacetic acid (400 ml). The reaction mixture was
stirred for 1 h at room temperature. After completion of the reaction, the
31.2, 24.6; MALDI-TOF-MS: Calcd For C17H18N2NaO6S: m/z 401.1. Found: mixture was concentrated and purified on silica gel column chromatography
401.1[MꢂNa]ꢂ.
(chloroform : methanolꢀ10 : 1) to give 22 (273 mg, 75%). [a]D25 ꢂ97.6 (cꢀ
1
4-[Benzyloxycarbonylpropyl-(4-nitro-benzenesulfonyl)-amino]-bu- 1.24, CHCl3); H-NMR (500 MHz, CDCl3) d: 8.08—7.02 (80H, m, 16Ph),

October 2009
1087
1
6.28 (1H, br s, CH2NHCH2), 6.01 (2H, d, J3ꢁ,4ꢁꢀ3.7 Hz, H-4ꢁ), 5.82 (2H, d, MeOH); H-NMR (500 MHz, CD3OD) d: 4.30 (2H, d, J1ꢁ,2ꢁꢀ7.9 Hz, H-1ꢁ),
J
3,4ꢀ3.1 Hz, H-4), 5.69—5.57 (6H, m, H-2,2ꢁ, H-3ꢁ), 5.49 (2H, dd, H-3), 4.21—4.17 (6H, m, H-1, 2POCH2), 3.07 (18H, s, 2N(CH3)3); 13C-NMR
4.81 (2H, d, J1ꢁ,2ꢁꢀ7.9 Hz, H-1ꢁ), 4.59 (2H, d, J1,2ꢀ8.0 Hz, H-1), 4.09—4.07 (125 MHz, CD3OD) d: 178.8, 175.2, 174.7, 105.2, 104.9, 75.4, 75.3, 75.0,
(4H, m, H-5ꢁ, H-6ꢁa), 3.98 (2H, t, H-5), 3.82—3.80 (2H, m, H-6ꢁb), 3.62— 74.8, 74.7, 72.6, 72.4, 70.9, 70.04, 69.98, 69.25, 67.6, 66.1, 60.58, 6055,
3.59 (6H, m, H-6, O–CH2), 3.06—2.83 (2H, m, O–CH2), 2.82—2.33 (4H, 54.8, 48.1, 47.0, 42.5, 40.5, 40.4, 34.4, 33.7, 33.0, 30.9, 30.7, 30.6, 30.5,
m, CONHCH2), 2.30 (4H, t, CH2NHCH2), 1.90 (4H, t, NHCOCH2), 1.92— 30.4, 30.3, 28.6, 27.9, 27.8, 26.8, 26.7, 25.1, 23.7, 14.5, 1.5; MALDI-TOF-
1.89 (4H, m, NHCOCH2CH2CH2N), 1.26—0.88 (34H, m, 2t-butyl, alkyl); MS: Calcd for C84H164N5O31P2: m/z 1801. Found: 1801 [MꢂH]ꢂ.
13C-NMR (125 MHz, CDCl3) d: 172.4, 165.6, 165.4, 165.3, 165.2, 165.1,
Glycocluster 27 Compound 27 was prepared from 20 (15 mg, 50 mmol)
135.5, 135.4, 133.3, 133.2, 133.13, 133.08, 132.7, 132.5, 130.0, 129.8, and 22 (346 mg, 0.13 mmol) as described for preparation of 21, yielding
129.74, 129.68, 129.63, 129.56, 129.5, 129.4, 129.1, 129.0, 128.9, 128.5, 287 mg (99%). [a]D25 ꢂ84.0 (cꢀ3.2, CHCl3); 1H-NMR (500 MHz, CDCl3)
128.4, 128.2, 127.7, 127.5, 101.4, 101.1, 73.6, 73.1, 71.8, 71.6, 70.1, 69.94, d: 7.98—6.95 (160H, m, 32Ph), 5.93 (4H, d, H-4ꢁ), 5.75 (4H, d, H-4),
69.88, 68.6, 67.7, 67.6, 60.6, 48.1, 39.5, 34.0, 29.1, 26.5, 25.5, 23.5, 18.8;
MALDI-TOF-MS: Calcd for C160H165N3NaO36Si2: m/z 2783.1. Found: (4H, d, J1ꢁ,2ꢁꢀ7.3 Hz, H-1ꢁ), 4.51 (4H, d, J1,2ꢀ7.9 Hz, H-1), 4.02—4.00 (8H,
2785.4 [MꢂNa]ꢂ.
m, H-5ꢁ, H-6ꢁa), 3.89 (4H, t, H-5), 3.73—3.70 (4H, m, H-6ꢁb), 3.53—3.51
5.61—5.54 (8H, m, H-2,2ꢁ), 5.51 (4H, dd, H-3ꢁ), 5.41 (4H, dd, H-3), 4.73
Glycocluster 23 To a solution of 22 (273 mg, 99 mmol) and 2-(tetrade- (12H, m, H-6, O–CH2), 3.25—3.12 (16H, m, O–CH2, CH2NCH2), 2.95—
cyl) hexadecanoic acid (113 mg, 0.24 mmol) in CH2Cl2 (2.0 ml) were added 2.93 (8H, m, 4CONHCH2), 2.22—2.05 (12H, m, 6NHCOCH2), 1.85—1.78
EDC (114 mg, 0.59 mmol) and DMAP (54 mg, 0.44 mmol). The reaction
(12H, m, 6NHCOCH2CH2CH2N), 1.26—0.78 (78H, m, 5t-butyl, alkyl); 13C-
mixture was stirred for 18 h at room temperature. After completion of the re- NMR (125 MHz, CDCl3) d: 171.3, 165.6, 165.4, 165.3, 165.24, 165.19,
action, the mixture was extracted with chloroform, washed with water, dried 165.1, 155.7, 135.5, 135.4, 133.31, 133.25, 133.1, 132.7, 132.5, 130.8,
(Na2SO4), and concentrated. The product was purified on silica gel column
130.0, 129.9, 129.8, 129.71, 129.66, 129.6, 129.5, 129.1, 129.0, 128.9,
128.8, 128.5, 128.4, 128.3, 128.2, 127.7, 127.5, 101.4, 101.1, 79.4, 73.6,
chromatography (toluene : acetoneꢀ5 : 1) to give 23 (276 mg, 87.3%). [a]D25
ꢂ76.6 (cꢀ5.7, CHCl3); 1H-NMR (500 MHz, CDCl3) d: 8.05—7.03 (80H, 73.1, 73.0, 71.9, 71.6, 70.1, 69.9, 68.6, 68.1, 67.7, 67.6, 60.7, 39.7, 39.5,
m, 16Ph), 6.01 (2H, J3ꢁ,4ꢁꢀ3.7 Hz, d, H-4ꢁ), 5.82 (2H, d, J3,4ꢀ3.2 Hz, H-4),
38.7, 32.8, 31.9, 30.3, 30.0, 29.7, 29.3, 29.1, 28.9, 28.5, 26.5, 25.54, 25.46,
5.69—5.63 (4H, m, H-2, 2ꢁ), 5.59 (2H, dd, H-3ꢁ), 5.51—5.47 (2H, m, H-3), 24.4, 23.9, 23.7, 22.9, 22.7, 18.8, 14.1, 10.9, 1.0; MALDI-TOF-MS: Calcd
4.81 (2H, d, J1ꢁ,2ꢁꢀ7.9 Hz, H-1ꢁ), 4.59 (2H, d, J1,2ꢀ7.9 Hz, H-1), 4.09—4.07 for C333H349N7NaO76Si4: m/z 5796.2. Found: 5795.5 [MꢂNa]ꢂ.
(4H, m, H-5ꢁ, H-6ꢁa), 3.97 (2H, t, H-5) 3.82—3.78 (2H, m, H-6ꢁb), 3.62—
Glycocluster 28 Compound 28 was prepared from 27 (287 mg,
3.59 (6H, m, H-6, O–CH2), 3.36—3.30 (4H, m, CH2NCH2), 3.24—3.08 50 mmol) as described for preparation of 22, yielding 266 mg (94%).
(2H, m, O–CH2), 3.08—3.00 (4H, m, CONHCH2), 2.61 (1H, br t, COCH),
2.16—2.14 (4H, m, NHCOCH2), 1.83 (4H, m, NHCOCH2CH2CH2N),
[a]D25 ꢂ87.6 (cꢀ1.9, CHCl3); 1H-NMR (500 MHz, CDCl3) d: 7.97—
6.95 (160H, m, Ar-H), 5.93 (4H, br s, H-4ꢁ), 5.75 (4H, br d, H-4),
1.30—0.80 (92H, m, 2t-butyl, alkyl); 13C-NMR (125 MHz, CDCl3) d: 171.1, 5.61—5.52 (12H, m, H-2,2ꢁ,3ꢁ), 5.50—5.41 (4H, m, H-3), 4.23 (4H, dꢃ2,
165.6, 165.4, 165.3, 165.24, 165.15, 165.1, 135.5, 135.4, 133.2, 133.1, J1ꢁ,2ꢁꢀ7.9 Hz, H-1ꢁ), 4.51 (4H, d, J1,2ꢀ7.9 Hz, H-1), 4.02—4.00 (8H, m,
132.7, 132.5, 130.0, 129.9, 129.8, 129.7, 129.6, 129.5, 129.1, 129.0, 128.9,
128.5, 128.4, 128.3, 128.2, 127.7, 127.5, 125.3, 101.4, 101.1, 73.6, 73.1,
73.0, 71.9, 71.6, 70.1, 70.0, 69.9, 68.6, 67.7, 67.6, 60.7, 47.3, 45.1, 41.6,
H-5ꢁ, H-6ꢁa), 3.90 (4H, t, H-5) 3.74—3.68 (4H, m, H-6ꢁb), 3.57—3.52
(12H, m, H-6, O–CH2), 3.25—3.12 (16H, m, O–CH2, CH2NCH2) 2.92
(10H, m, CONHCH2, CH2NHCH2), 2.49 (4H, br, CH2NHCH2CH2CH2CO)
39.7, 39.4, 33.2, 32.9, 31.9, 30.0, 29.6, 29.5, 29.32, 29.285, 29.14, 29.08, 2.07—2.03 (8H, m, NHCOCH2CH2CH2NHCOCH2), 1.89—1.73 (12H, m,
27.8, 26.5, 25.6, 25.5, 25.0, 24.1, 22.7, 18.8, 14.1; MALDI-TOF-MS: Calcd
6CONHCH2CH2CH2-CONH) 1.18—0.63 (68H, m, 4t-butyl, alkyl); 13C-NMR
(125 MHz, CDCl3) d: 172.8, 172.2, 171.6, 165.6, 165.4, 165.3, 1652,
for C190H223N3NaO37Si2: m/z 3217. Found: 3217 [MꢂNa]ꢂ.
Glycocluster 24 Compound 24 was prepared from 23 (228 mg) as de- 165.1, 135.5, 135.4, 133.33, 133.26, 133.1, 132.7, 132.5, 130.0, 129.9,
scribed for preparation of 4, yielding 101 mg (52%). [a]D25 ꢂ105.5 (cꢀ0.7, 129.8, 129.71, 129.66, 129.6, 129.5, 129.1, 129.0, 128.9, 128.5, 128.4,
CHCl3); 1H-NMR (500 MHz, CDCl3) d: 8.11—7.02 (6H, m, 12Ph), 5.93 128.3, 128.2, 127.7, 127.5, 101.4, 101.1, 73.6, 73.0, 71.9, 71.6, 70.1, 69.9,
(2H, br d, H-4ꢁ), 5.82—5.78 (4H, m, H-2ꢁ, 4), 5.71—5.67 (2H, m, H-2), 68.6, 67.7, 67.6, 60.7, 48.0, 47.3, 45.1, 39.4, 33.3, 32.6, 31.9, 30.8, 29.7,
5.56—5.52 (4H, m, H-3, 3ꢁ), 4.88 (2H, d, J1ꢁ,2ꢁꢀ7.3 Hz, H-1ꢁ), 4.64 (2H, d, 29.3, 29.2, 29.1, 26., 25.5, 24.3, 23.6, 22.7, 18.8, 14.1, 1.0; MALDI-TOF-
J
1,2ꢀ7.3 Hz, H-1), 4.14—4.10 (4H, m, H-5ꢁ, H-6ꢁa), 3.98—3.91 (4H, m, H-
MS: Calcd for C328H341N7NaO74Si4: m/z 5696. Found: 5696 [MꢂNa]ꢂ.
Glycocluster 29 Compound 29 was prepared from 28 (344 mg) as de-
scribed for preparation of 23, yielding 331 mg (90%). [a]D25 ꢂ86.2 (cꢀ5.7,
5, 6ꢁb), 3.72—3.65 (4H, m, H-6a, O–CH2), 3.51—3.47 (2H, m, H-6b), 3.34
(6H, br, O–CH2, CH2NCH2), 3.07 (4H, br, CONHCH2), 2.64 (1H, br t,
COCH), 2.16 (4H, br, NHCOCH2), 1.85 (4H, br, NHCOCH2CH2CH2N),
1
CHCl3); H-NMR (500 MHz, CDCl3) d: 7.97—6.95 (160H, m, 32Ph), 5.93
1.56—0.86 (74H, m, alkyl); 13C-NMR (125 MHz, CDCl3) d: 171.3, 166.5, (4H, d, H-4ꢁ), 5.75 (4H, d, H-4), 5.61—5.50 (12H, m, H-2,2ꢁ,3ꢁ), 5.41 (4H,
165.5, 165.2, 133.6, 166.5, 165.2, 133.6, 133.5, 133.2, 133.1, 130.1, 129.9,
dd, H-3), 4.73 (4H, dꢃ2, J1ꢁ,2ꢁꢀ7.6 Hz, H-1ꢁ), 4.51 (4H, d, J1,2ꢀ7.9 Hz, H-
129.64, 129.59, 129.4, 129.3, 129.1, 128.8, 128.71, 128.67, 128.6, 128.5, 1), 4.02—4.00 (8H, m, H-5ꢁ, H-6ꢁa), 3.89 (4H, t, H-5) 3.74—3.68 (4H, m,
128.34, 128.29, 128.23, 128.15, 101.4, 101.3, 74.1, 73.1, 71.8, 70.0, 69.9,
68.7, 68.6, 67.9, 60.5, 47.2, 45.2, 41.5, 39.4, 33.2, 33.0, 31.8, 29.9, 29.6,
H-6ꢁb), 3.57—3.52 (12H, m, H-6, O–CH2), 3.25—3.12 (16H, m, O–CH2,
CH2NCH2), 2.95—2.92 (10H, m, CONHCH2, CH2NHCH2), 2.26—2.23 (4H,
29.5, 29.3, 29.1, 27.7, 26.5, 25.5, 25.4, 25.1, 24.1, 22.6, 14.0; MALDI-TOF- br, CH2NHCH2CH2CH2CO), 2.05 (8H, br s, NHCOCH2CH2CH2NCOCH2),
MS: Calcd for C158H187N3NaO37: m/z 2741. Found: 2741 [MꢂNa]ꢂ.
1.85—1.75 (12H, br s, COCH2CH2CH2N), 1.17—0.71 (126H, m, 4t-butyl,
Glycocluster 25 Compound 25 was prepared from 24 (101 mg) as de- alkyl); 13C-NMR (125 MHz, CDCl3) d: 176.5, 172.6, 172.3, 171.9, 171.4,
scribed for preparation of 8, yielding 61 mg (54%). [a]D25 ꢂ76.4 (cꢀ1.0, 171.3, 167.8, 165.6, 165.4, 165.3, 165.24, 165.18, 165.1, 135.5, 135.4,
1
MeOH); H-NMR (500 MHz, CD3OD) d: 8.07—7.20 (60H, m, 12Ph), 5.93 133.3, 133.1, 132.7, 132.5, 133.1, 132.5, 130.0, 129.9, 129.8, 129.71,
(2H, d, J3ꢁ,4ꢁꢀ3.7 Hz, H-4ꢁ), 5.89 (2H, d, J3,4ꢀ3.1 Hz, H-4), 5.73—5.60 (8H, 129.68, 129.61, 129.5, 129.4, 129.1, 129.0, 128.9, 128.5, 128.5, 128.4,
m, H-2, 2ꢁ, 3, 3ꢁ), 5.10 (2H, d, J1ꢁ,2ꢁꢀ7.3 Hz, H-1ꢁ), 4.81 (2H, d, J1,2ꢀ7.9 Hz,
128.3, 128.2, 127.7, 127.5, 126.4, 101.5, 101.1, 77.4, 77.1, 76.8, 73.6, 73.0,
H-1), 4.35—4.33 (4H, m, H-5ꢁ, H-6ꢁa), 4.26 (4H, br s, POCH2CH2), 4.17— 71.9, 71.6, 70.1, 69.9, 68.6, 67.6, 60.7, 47.4, 47.3, 47.2, 45.7, 45.1, 44.7,
4.14 (2H, m, H-5ꢁ), 3.97 (2H, br, H-6a), 3.89—3.81 (4H, m, H-6b, 6ꢁb), 41.4, 39.4, 33.3, 32.9, 32.7, 31.9, 30.9, 30.4, 30.1, 29.7, 29.6, 29.3, 29.2,
3.63—3.51 (6H, m, POCH2CH2, OCH2), 3.37—3.33 (6H, m, O–CH2, 27.8, 26.5, 25.54, 25.47, 24.9, 24.4, 23.9, 23.6, 22.7, 18.8, 14.1; MALDI-
CH2NCH2), 3.17 (18H, s, N(CH3)3), 3.00—2.95 (4H, m, NHCH2), 2.66 TOF-MS: Calcd for C358H399N7NaO75Si4: m/z 6131. Found: 6131 [MꢂNa]ꢂ.
(1H, br t, COCH), 2.17 (4H, tꢃ2, NCH2CH2CH2CO), 1.88—1.78 (4H,
m, NCH2CH2CH2CO), 1.54—0.83 (74H, m, alkyl); 13C-NMR (125 MHz,
CD3OD) d: 178.7, 175.0, 174.5, 167.1, 167.0, 166.9, 166.8, 166.7, 134.9,
Glycocluster 30 Compound 30 was prepared from 29 (331 mg) as de-
scribed for preparation of 24, yielding 237 mg (85%). [a]D25 ꢂ110.5 (cꢀ2.6,
1
CHCl3); H-NMR (500 MHz, CDCl3) d: 8.10—7.20 (120H, m, 24Ph), 5.93
134.8, 134.6, 134.5, 130.92, 130.89, 130.71, 130.65, 130.6, 130.5, 130.3, (4H, br s, H-4ꢁ), 5.82—5.78 (8H, m, H-2ꢁ, 4), 5.69 (4H, br t, H-2), 5.57—
130.2, 129.91, 129.87, 129.7, 129.6, 129.5, 129.4, 102.2, 73.6, 73.4, 73.3,
73.0, 71.7, 71.4, 70.7, 70.2, 69.4, 68.7, 64.1, 60.8, 54.7, 49.5, 49.3, 49.2,
49.12, 49.07, 48.9, 48.8, 48.73, 48.66, 48.5, 48.3, 47.0, 42.6, 40.4, 40.3,
34.5, 34.4, 33.7, 33.0, 31.0, 30.8, 30.7, 30.64, 30.57, 30.4, 30.3, 30.2,
5.52 (8H, m, H-3,3ꢁ), 4.90 (4H, d, J1ꢁ,2ꢁꢀ7.9 Hz, H-1ꢁ), 4.65 (4H, d, J1,2ꢀ
7.9 Hz, H-1), 4.16—4.10 (8H, m, H-5ꢁ, H-6ꢁa), 3.99—3.90 (8H, m, H-5,
6ꢁb), 3.72—3.64 (8H, m, H-6a, O–CH2), 3.51—3.44 (4H, m, H-6b), 3.34—
3.25 (16H, m, 4O–CH2, 4CH2NCH2), 3.05 (8H, br, CONHCH2), 2.62 (1H,
28.7, 27.7, 27.6, 26.7, 25.2, 23.7, 14.5; MALDI-TOF-MS: Calcd for br t, COCH), 2.35—2.32 (4H, m, CH2NHCH2CH2CH2CO), 2.14 (8H, br,
C168H212N5O43P2: m/z 3049. Found: 3049 [MꢂH]ꢂ.
NHCOCH2CH2CH2NHCOCH2), 1.84—1.75 (12H, br s, COCH2CH2CH2NH),
Glycocluster 26 (B) Compound 26 was prepared from 25 (39 mg) as 1.52—0.85 (90H, m, alkyl); 13C-NMR (125 MHz, CDCl3) d: 176.6, 172.8,
described for preparation of 13, yielding 22 mg (84%). [a]D25 ꢄ11.1 (cꢀ0.3, 172.7, 172.5, 172.0, 171.7, 171.5, 166.5, 165.6, 165.5, 165.3, 133.7, 133.6,

1088
Vol. 57, No. 10
133.3, 133.2, 130.1, 130.0, 129.73, 129.66, 129.5, 129.4, 129.2, 128.9,
128.8, 128.6, 128.44, 128.39, 128.33, 128.25, 127.8, 101.5, 101.3, 74.1,
73.2, 71.9, 71.8, 70.1, 70.0, 69.0, 68.73, 68.66, 67.9, 60.5, 47.4, 47.3, 45.1,
44.8, 41.5, 39.5, 33.3, 33.1, 32.8, 31.9, 30.5, 30.1, 29.72, 29.67, 29.6, 29.4,
29.2, 27.8, 26.5, 25.6, 25.5, 24.9, 24.6, 24.5, 23.9, 23.6, 22.7, 14.1; MALDI-
TOF-MS: Calcd for C294H327N7O75Na: m/z 5178. Found: 5178 [MꢂNa]ꢂ.
Glycocluster 31 Compound 31 was prepared from 30 (133 mg) as de-
scribed for preparation of 8, yielding 31 mg (21%). [a]D25 ꢂ79.6 (cꢀ0.8,
MeOH); 1H-NMR (500 MHz, CD3OD) d: 8.03—7.18 (120H, m, 24H), 5.94
(4H, d, H-4ꢁ), 5.88 (4H, d, H-4), 5.73—5.59 (16H, m, H-3, 3ꢁ, 2, 2ꢁ), 5.04
(4H, d, J1ꢁ,2ꢁꢀ6.7 Hz, H-1ꢁ), 4.82 (4H, d, J1,2ꢀ7.9 Hz, H-1), 4.37—4.32
(8H, m, H-5, H-6ꢁa), 4.20—4.14 (8H, m, PO–CH2, H-5ꢁ), 4.00—3.95 (4H,
4) Hada N., Kuroda M., Takeda T., Chem. Pharm. Bull., 48, 1160—1165
(2000).
5) Hada N., Ohtsuka I., Sugita M., Takeda T., Tetrahedron Lett., 41,
9065—9068 (2000).
6) Hada N., Sato K., Sakushima J.-I., Goda Y., Sugita M., Takeda T.,
Chem. Pharm. Bull., 49, 1464—1467 (2001).
7) Ohtsuka I., Hada N., Ohtaka H., Sugita M., Takeda T., Chem. Pharm.
Bull., 50, 600—604 (2002).
8) Ohtsuka I., Hada N., Sugita M., Takeda T., Carbohydr. Res., 337,
2037—2047 (2003).
9) Yamamura T., Hada N., Kaburaki A., Yamano K., Takeda T., Carbo-
hydr. Res., 339, 2749—2759 (2004).
m, H-6a), 3.88—3.77 (8H, m, H-6b, 6ꢁb), 3.57 (12H, t, PO–CH2, OCH2), 10) Hada N., Sonoda Y., Takeda T., Carbohydr. Res., 341, 1341—1352
3.38—3.30 (20H, m, 4O–CH2, 3CH2NCH2), 3.17 (36H, s, N(CH3)3), 2.96—
2.95 (8H, m, CONHCH2), 2.61 (1H, br t, COCH), 2.42—2.33 (4H, m,
CH2NHCH2CH2CH2CO), 2.18—2.11 (8H, m, NHCOCH2CH2CH2NCOCH2),
(2006).
11) Hada N., Nakashima T., Shrestha S. P., Masui R., Narukawa Y., Tani
K., Takeda T., Bioorg. Med. Chem. Lett., 17, 5912—5915 (2007).
1
.82—1.76 (12H, m, COCH2CH2CH2NH), 1.51—0.81 (90H, m, alkyl); 13C- 12) Hada N., Shida Y., Shimamura H., Sonooda Y., Kasahara T., Sugita
NMR (125 MHz, CD3OD) d: 181.2, 167.1, 167.0, 166.9, 166.8, 166.7,
134.9, 134.8, 134.63, 134.60, 134.56, 134.5, 130.9, 130.7, 130.63, 130.55,
130.5, 130.3, 130.2, 129.9, 129.7, 129.5, 102.2, 102.0, 73.6, 73.5, 73.3,
M., Takeda T., Carbohydr. Res., 343, 2221—2228 (2008).
13) Sugita M., Fujii H., Dulaney T. J., Inagaki F., Suzuki M., Suzuki A.,
Ohta, S., Biochim. Biophys. Acta, 1259, 220—226 (1995).
71.7, 71.5, 70.7, 70.3, 69.5, 68.9, 67.5, 67.4, 63.9, 60.5, 60.4, 54.7, 40.38, 14) Kean D. E., Ohtsuka I., Sato K., Hada N., Takeda T., Lochnit G.,
40.35, 33.0, 31.0, 30.8, 30.7, 30.6, 30.43, 30.36, 30.3, 28.7, 27.7, 26.7, 23.7,
14.5; MALDI-TOF-MS: Calcd for C314H375N11O87P4Na: m/z 5836. Found:
5836 [MꢂNa]ꢂ.
Geyer R., Harnett M. M., Harnett W., Parasite Immunol., 28, 69—76
(2006).
15) Sauter N. K., Bednarski M. D., Wurzberg B. A., Hanson J. E., White-
sides G. M., Skehel J. J., Wiley D. C., Biochemistry, 28, 8388—8396
(1989).
Glycocluster 32 (C) Compound 32 was prepared from 31 (31 mg) as
described for preparation of 13, yielding 14 mg (80%). [a]D25 ꢄ8.95 (cꢀ0.4,
1
MeOH); H-NMR (500 MHz, CD3OD) d: 4.30 (4H, d, J1ꢁ,2ꢁꢀ7.9 Hz, H-1ꢁ), 16) Lee R. T., Lee Y. C., Glycoconjug. J., 17, 543—551 (2000).
4.21 (4H, d, J1ꢁ,2ꢁꢀ7.3 Hz, H-1), 4.16 (8H, br s, 4POCH2), 3.06 (36H, s, 17) Sato K., Hada N., Takeda T., Carbohydr. Res., 341, 836—845 (2006).
4N(CH3)3); 13C-NMR (125 MHz, CD3OD) d: 178.7, 175.3, 175.2, 174.7, 18) Patel A., Lindhorst T. K., J. Org. Chem., 66, 2674—2680 (2001).
174.6, 174.2, 105.3, 105.0, 75.40, 75.35, 75.1, 74.84, 74.75, 72.6, 72.4,
70.9, 70.1, 69.3, 67.6, 66.1, 60.61, 60.58, 54.8, 49.8, 49.6, 42.5, 40.5,
34.5, 34.4, 33.8, 33.6, 33.1, 31.4, 31.0, 30.8, 30.6, 30.5, 30.4, 28.7, 27.9,
26.8, 25.9, 25.1, 25.0, 24.7, 23.7, 14.5; MALDI-TOF-MS: Calcd for
C146H280N11O63P4: m/z 3319.8. Found: 3320.3 [MꢂH]ꢂ.
19) Seebach D., Beck A. T., Bierbaum D. J., Chemistry & Biodiversity, 8,
1111—1239 (2004).
20) Nicolaou K. C., Bockovich N. J, Carcanague D. R., J. Am. Chem. Soc.,
115, 8843—8844 (1993).
21) Veeneman G. H., van Leeuwen S. H., van Boom J. H., Tetrahedron
Lett., 31, 1331—1334 (1990).
Acknowledgement This work was supported by a Grant-in-Aid for Sci- 22) Konradsso P., Udodong U. E., Fraser-Reid B., Tetrahedron Lett., 31,
entific Research (No. 19590011) from the Ministry of Education, Culture, 4313—4316 (1990).
Sports, Science and Technology of Japan (MEXT), and Open Research Cen- 23) Pekari K., Tailler D., Weingart R., Schmidt R. R., J. Org. Chem., 66,
ter project of the MEXT.
7432—7442 (2001).
24) Nishida Y., Takamori Y., Matsuda K., Ohrui H., Yamada T., Kobayashi
K., J. Carbohydr. Chem., 18, 985—997 (1999).
References
1) Hada N., Takeda T., Ogihara Y., Carbohydr. Res., 258, 93—104 25) Mickeleit M., Wieder T., Arnold M., Geilen C. C., Mulzer J., Reutter
(1994).
W., Angew. Chem. Int. Ed., 37, 351—353 (1998).
26) Mitsunobu O., Synthesis, 1981, 1—28 (1981).
2) Hada N., Hayashi E., Takeda T., Carbohydr. Res., 316, 58—70 (1999).
3) Hada N., Matsuzaki A., Takeda T., Chem. Pharm. Bull., 47, 1265— 27) Fukuyama T., Jow C.-K., Cheung M., Tetrahedron Lett., 36, 6373—
1268 (1999). 6374 (1995).