First Synthesis of a Trisaccharide of Glycosylkaemferide: A Resistance Factor in Carnations

Article abstract of DOI:10.1081/SCC-120027259

A trisaccharide, phenyl β-D-glucopyranosyl-(1→2)-[α -L-rhamnopyranosyl-(1→6)]-1-thio-β-D-glucopyranoside, of glycosylkaemferide, a resistance factor in carnations, was synthesized in a practical way.

Full text of DOI:10.1081/SCC-120027259

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First Synthesis of a Trisaccharide of
Glycosylkaemferide: A Resistance Factor in
Carnations
Mamoru Koketsu a , Motoaki Kuwahara a , Hisako Sakurai a & Hideharu Ishihara a
a Department of Chemistry, Faculty of Engineering, Gifu University, Gifu, 501-1193,
Japan
Published online: 18 Oct 2011.
To cite this article: Mamoru Koketsu , Motoaki Kuwahara , Hisako Sakurai & Hideharu Ishihara (2004): First Synthesis of
a Trisaccharide of Glycosylkaemferide: A Resistance Factor in Carnations, Synthetic Communications: An International
Journal for Rapid Communication of Synthetic Organic Chemistry, 34:2, 239-245
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SYNTHETIC COMMUNICATIONSw
Vol. 34, No. 2, pp. 239–245, 2004
First Synthesis of a Trisaccharide of
Glycosylkaemferide: A Resistance
Factor in Carnations
*
Mamoru Koketsu, Motoaki Kuwahara, Hisako Sakurai,
*
and Hideharu Ishihara
Department of Chemistry, Faculty of Engineering, Gifu University,
Gifu, Japan
ABSTRACT
A trisaccharide, phenyl b-D-glucopyranosyl-(1 ! 2)-[a-L-rhamnopyra-
nosyl-(1 ! 6)]-1-thio-b-D-glucopyranoside, of glycosylkaemferide, a
resistance factor in carnations, was synthesized in a practical way.
Key Words: Trisaccharide; Carnation; Flavonol; Antifungal compound.
Many carnations (Dianthus caryophyllus) suffer from a fungal parasite,
Fusarium oxysporum f. sp. Dianthi, which causes severe crop losses world-
wide.[1] Carnations of the cultivar Novada bearing a high resistance to this
*
Correspondence: Mamoru Koketsu and Hideharu Ishihara, Department of Chemistry,
Faculty of Engineering, Gifu University, Gifu 501-1193, Japan.
239
DOI: 10.1081/SCC-120027259
0039-7911 (Print); 1532-2432 (Online)
www.dekker.com
Copyright # 2004 by Marcel Dekker, Inc.

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Koketsu et al.
Scheme 1. Retrosynthetic analysis.
parasite are of commercial interest.[2] Recently, kaempferide triglycoside has
been reported, for the first time, to be one of the most important resistance
factors in the carnation.[3] The total synthesis of the kaempferide triglycoside
is required for the detailed elucidation of its antifungal mechanism. In this
paper, we describe the first preparative synthesis of the trisaccharide, phenyl
2,3,4,6-tetra-O-acetyl-b-D-glucopyranosyl-(1 ! 2)-[2,3,4-tri-O-acetyl-a-L-
rhamnopyranosyl-(1 ! 6)]-4-O-acetyl-3-O-benzyl-1-thio-b-D-glucopyrano-
side 1, a critical portion of this antifungal compound (Sch. 1).
RESULTS AND DISCUSSION
Retrosynthetic analysis indicates that the trisaccharide can be obtained by
condensation of a rhamnose donor and a glucose dimer acceptor. The
disaccharide could be constructed from a glucose donor and a 2-hydroxy-
glucose acceptor (Sch. 1).
2-Hydroxy-glucose acceptor 5 was prepared from 1,2:5,6-di-O-isopropy-
lidene-a-D-glucofuranose in seven steps as shown in Sch. 2. 1,2:5,6-di-O-
isopropylidene-a-D-glucofuranose was benzylated, hydrolyzed with acid, and
treated with acetic anhydride-sodium acetate. 1,2,4,6-Tetra-O-acetyl-3-O-
benzyl-b-D-glucopyranose 2 was obtained in four steps in a satisfactory yield
(overall 80%).[4] A thiophenyl group was chosen as both the leaving and
protecting group at the reducing end of the glucose 3.[5] 2-Hydroxy-glucose
acceptor 5 was prepared from 3 through selective deacetylation 4 and 4,6-O-
benzylidenation of glucose.[6]
Synthesis of glucose donor 7 was achieved by peracetylation (quantitative
yield), followed by a regioselective hydrazinolysis[7] of the 1-O-acetyl group
of peracetylated glucose to afford 6 (87%). Subsequent treatment of 6 with
trichloroacetonitrile afforded trichloroacetimidate 7 (59%) (Sch. 3).

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First Synthesis of a Trisaccharide of Glycosylkaemferide
241
Scheme 2. Synthesis of glucose acceptor 5. Reagents and conditions: (a) BnBr, NaH,
DMF, 908C, 2 h; (b) 60% AcOH/H2O, 908C, 2 h; (c) 0.1 M H2SO4/H2O, 908C, 2 h;
.
(d) Ac2O, NaOAc, 1108C, 6 h, 80% (overall yield for 4 steps); (e) PhSH, Et2O BF3,
CH2Cl2, rt, overnight, 63%; (f) NaOEt, MeOH, rt, overnight, quant.; (g) PhCH(OMe)2,
p-MeC6H4SO3H, CH3CN, rt, 3 h, 82%.
The 2-hydroxy-D-glucose acceptor 5 was glycosylated with 7 in the
.
presence of Et2O BF3 to give (1 ! 2)-linked disaccharide 8. Removal of the
4,6-O-benzylidene group of 8 by acid hydrolysis[8] afforded 9 (58%, 2 steps
overall yield). The 1H and 13C NMR spectra of 9 showed that the reaction gave
exclusively the b-glycoside.
Rhamnose trichloroacetimidate donor 10[9] was prepared from L-
rhamnopyranose by the same procedures shown in Sch. 3 (3 steps in 70%
overall yield). Selective glycosylation of the 6-OH in 9 with 2,3,4-tri-O-
acetyl-a-L-rhamnopyranosyl trichloroacetimidate (10) gave the trisaccharide
derivative 11 in 54% yield. Total synthesis of the target trisaccharide moiety 1
was achieved by quantitative acetylation of 11 (Sch. 4).
Scheme 3. Synthesis of glucose donor 7. Reagents and conditions: (a) Ac2O,
pyridine, rt, overnight, quant.; (b) H2NNH2/AcOH, DMF, 508C, 2 h, 87% (2 steps);
(c) CCl3CN, DBU, CH2Cl2, 08C, 2 h, 59%.

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Koketsu et al.
Scheme 4. Synthesis of target trisaccharide 1. Reagents and conditions: (a)
.
Et2O BF3, CH2Cl2, 08C, 2 h; (b) 70% AcOH/H2O, 708C, 4 h, 58% (2 steps overall);
.
(c) Et2O BF3, CH2Cl2, 2208C, 5 h, 54%; (d) Ac2O, pyridine, rt, 12 h, quant.
EXPERIMENTAL
General Procedures
Melting points were determined by use of a Yanagimoto micromelting
1
point apparatus and are uncorrected. H and 13C spectra were recorded on a
JEOL JNM-a400 spectrometer. Mass spectra were obtained on Shimadzu
9020-DF mass spectrometer.
1,2,4,6-Tetra-O-acetyl-3-O-benzyl-b-D-glucopyranose[4] 2. M.p.:
1
106.2–107.28C; H NMR (CDCl3) d 1.98 (6H, s, 2CH3), 1.98 (3H, s, CH3),
2.08 (3H, s, CH3), 2.10 (3H, s, CH3), 3.75 (1H, m, H5), 3.78 (1H, t, J ¼ 9.2 Hz,
H3), 4.09 (1H, dd, J ¼ 12.0, 2.4 Hz, H6a), 4.23 (1H, dd, J ¼ 12.0, 4.8 Hz,
H6b), 4.62 (2H, s, CH2Ph), 5.15 (1H, t, J 9.2 Hz, H4), 5.16 (1H, t, J ¼ 8.4 Hz,
H2), 5.66 (1H, d, J ¼ 8.4 Hz, H1), 7.23–7.35 (5H, m, Ph); 13C NMR (CDCl3)
d 20.3, 20.4, 20.6, 21.9, 61.7, 69.0, 71.5, 73.0, 74.2, 79.9, 91.9, 127.5–137.5,
169.0, 169.19, 169.22, 170.7; CIMS: m/z 439 [Mþ þ 1].
Phenyl 2,4,6-tri-O-acetyl-3-O-benzyl-1-thio-b-D-glucopyranose[5] 3.
1
[a]24D 27.58 (c, 0.05, CHCl3); M.p.: 114.0–114.88C; H NMR (CDCl3) d
1.96 (3H, s, CH3), 2.04 (3H, s, CH3), 2.07 (3H, s, CH3), 3.61–3.66 (1H, m),
3.72 (1H, t, J 9.2 Hz), 4.17 (2H, m), 4.58 (1H, d, J ¼ 11.0 Hz), 4.61 (1H, d,
J ¼ 11.0 Hz), 4.64 (1H, d, J ¼ 10.0 Hz), 5.03–5.10 (2H, m), 7.21–7.50 (10H,
m); 13C NMR (CDCl3) d 20.69, 20.73, 20.9, 62.5, 69.6, 71.3, 74.2, 76.1, 81.5,
86.2, 169.2, 169.3, 170.6; CIMS: m/z 489 [Mþ þ 1].

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First Synthesis of a Trisaccharide of Glycosylkaemferide
243
Phenyl 3-O-benzyl-4,6-O-benzylidene-1-thio-b-D-glucopyranose 5.
1
[a]25D 260.08 (c, 0.05, CHCl3); M.p.: 127.5–128.08C; H NMR (CDCl3) d
2.58 (1H, d, J 2.4 Hz, OH), 3.47–3.71 (4H, m, H2–H5), 3.79 (1H, t,
J ¼ 10.2 Hz, H6a), 4.38 (1H, dd, J ¼ 10.2, 5.0 Hz, H6b), 4.62 (1H, d, J 9.2 Hz,
H1), 4.78 (1H, d, J ¼ 11.4 Hz, CH2Ph), 4.95 (1H, d, J ¼ 11.4 Hz, CH2Ph),
5.56 (1H, s, CHPh), 7.24–7.54 (15H, m, 3Ph); 13C NMR (CDCl3) d 68.6, 70.7,
72.2, 74.8, 81.1, 81.6, 88.4, 101.2, 126.0–138.1; CIMS: m/z 451 [Mþ þ 1].
2,3,4,6-Tetra-O-acetyl-a-D-glucopyranosyl trichloroacetimidate 7.
1
[a]27D þ52.08 (c, 0.001, CHCl3); H NMR (CDCl3) d 2.02 (3H, s, CH3),
2.04 (3H, s, CH3), 2.06 (3H, s, CH3), 2.08 (3H, s, CH3), 4.13 (1H, dd,
J ¼ 12.8, 2.0 Hz, H6a), 4.20–4.24 (1H, m, H5), 4.28 (1H, dd, J ¼ 12.4,
4.0 Hz, H6b), 5.14 (1H, dd, J ¼ 10.0, 4.0 Hz, H2), 5.19 (1H, t, J ¼ 10.0 Hz,
H3), 5.57 (1H, t, J ¼ 9.6 Hz, H-4), 6.57 (1H, d, J ¼ 4.0 Hz, H1), 8.71 (1H, s,
NH); 13C NMR (CDCl3) d 20.4, 20.5, 20.6, 61.3, 67.7, 69.6, 69.8, 69.9, 92.8,
160.7, 169.4, 169.8, 169.9, 170.5; CIMS: m/z 492 [Mþ þ 1].
Phenyl
2,3,4,6-tetra-O-acetyl-b-D-glucopyranosyl-(1 ! 2)-3-O-
benzyl-1-thio-b-D-glucopyranoside 9. [a]26D 220.08 (c, 0.001, CHCl3);
M.p.: 65.5–68.58C; 1H NMR (CDCl3) d 2.01 (3H, s, CH3), 2.02 (3H, s, CH3),
3.29 (1H, m), 3.44 (1H, t, J ¼ 8.8 Hz,), 3.64 (1H, t, J ¼ 9.2 Hz,), 3.71–3.84
(3H, m), 4.09–4.14 (3H, m), 4.22 (1H, dd, J ¼ 12.4, 4.4 Hz), 4.67 (1H, d,
J ¼ 9.6 Hz) 4.83 (2H, s, CH2Ph), 5.10–5.18 (3H, m), 7.24–7.49 (10H, m, Ph);
13C NMR (CDCl3) d 20.5, 20.7, 21.0, 61.7, 62.2, 68.3, 70.8, 71.6, 71.9, 73.1,
75.6, 76.5, 79.0, 85.7, 86.7, 99.6, 127.4–137.9 (Ph), 169.1, 169.3, 170.2,
170.7; FABMS (pos.-ion): m/z 693 [Mþ þ 1].
2,3,4-Tri-O-acetyl-a-L-rhamnopyranosyl trichloroacetimidate[9] 10.
[a]27D 272.08 (c, 0.001, CHCl3); 1H NMR (CDCl3) d 1.27 (3H, d, J ¼ 5.6 Hz,
CH3), 2.00 (3H, s, CH3), 2.08 (3H, s, CH3), 2.19 (3H, s, CH3), 4.10 (1H, m,
H5), 5.18 (1H, t, J ¼ 10.4 Hz, H4), 5.37 (1H, dd, J ¼ 10.4, 3.2 Hz, H3), 5.46
(1H, dd, J ¼ 3.2, 2.0 Hz, H2), 6.20 (1H, d, J ¼ 2.0 Hz, H1) 8.74 (1H, s, NH);
13C NMR (CDCl3) d 17.4, 20.6, 20.7, 68.0, 68.7, 69.2, 70.2, 90.6, 94.6, 159.9,
169.75, 169.82; CIMS: m/z 435 [Mþ þ 1].
Phenyl 2,3,4,6-tetra-O-acetyl-b-D-glucopyranosyl-(1 ! 2)-[2,3,4-tri-
O-acetyl-a-L-rhamnopyranosyl-(1 ! 6)]-3-O-benzyl-1-thio-b-D-gluco-
pyranoside 11. [a]26D 256.08 (c, 0.001, CHCl3); M.p.: 80.2–81.08C; H
1
NMR (CDCl3) d 1.18 (3H, d, J 6.0 Hz, CH3), 2.01 (3H, s, CH3), 2.02 (3H, s,
CH3), 2.03 (3H, s, CH3), 2.04 (3H, s, CH3), 2.06 (3H, s, CH3), 2.09 (3H, s,
CH3), 2.14 (3H, s, CH3), 3.40 (1H, m) 3.52 (1H, t, J ¼ 8.8 Hz), 3.60–3.70
(3H, m), 3.82 (1H, t, J ¼ 8.8 Hz) 3.89–3.95 (2H, m), 4.06 (1H, dd, J ¼ 12.4,
2.4 Hz) 4.12 (1H, dd, J ¼ 14.4, 7.2 Hz), 4.26 (1H, dd, J ¼ 12.0, 4.4 Hz) 4.62
(1H, d, J ¼ 9.2 Hz), 4.73 (1H, s) 4.77 (1H, d, J ¼ 10.4 Hz, CH2Ph), 4.88 (1H,
d, J ¼ 11.2 Hz, CH2Ph) 5.01–5.20 (5H, m), 5.23–5.26 (2H, m) 7.22–7.52
(10H, m, Ph); 13C NMR (CDCl3) d 20.6, 20.8, 20.8, 20.9, 61.8, 66.5, 66.8,

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Koketsu et al.
68.3, 69.2, 69.4, 70.1, 70.8, 71.7, 72.0, 73.2, 75.8, 76.5, 78.0, 85.8, 86.9, 97.7,
99.6, 127–137.8,169.2, 169.4, 167.0, 170.1, 107.1, 170.3, 170.7; FABMS
(pos.-ion): m/z 965 [Mþ þ 1].
Phenyl 2,3,4,6-tetra-O-acetyl-b-D-glucopyranosyl-(1 ! 2)-[2,3,4-tri-
O-acetyl-a-L-rhamnopyranosyl-(1 ! 6)]-4-O-acetyl-3-O-benzyl-1-thio-
b-D-glucopyranoside 1. [a]26D 249.08 (c, 0.001, CHCl3); M.p.: 73.0–
75.08C; 1H NMR (CDCl3) d 1.19 (3H, d, J ¼ 6.4 Hz, CH3), 1.97 (3H, s, CH3),
1.99 (3H, s, CH3), 2.01 (3H, s, CH3), 2.02 (3H, s, CH3), 2.04 (3H, s, CH3), 2.06
(3H, s, CH3), 2.09 (3H, s, CH3), 2.12 (3H, s, CH3) 3.53–3.61 (3H, m), 3.64–
3.71 (2H, m), 3.81 (1H, dd, J ¼ 9.4, 6.2 Hz) 3.88 (1H, t, J ¼ 9.0 Hz), 4.06 (1H,
dd, J ¼ 12.6, 2.2 Hz) 4.24 (1H, dd, J ¼ 12.6, 4.6 Hz), 4.60 (1H, d,
J ¼ 11.2 Hz) 4.63 (1H, d, J ¼ 10.0 Hz), 4.72 (1H, s) 4.79 (1H, d,
J ¼ 10.8 Hz,), 4.95–5.25 (8H, m) 7.22–7.51 (10H, m, Ph); 13C NMR
(CDCl3) d 20.5–20.9, 61.8, 66.5, 67.1, 68.2, 69.0, 69.4, 70.8, 70.9, 71.8, 71.9,
73.2, 75.7, 76.2, 84.4, 85.7, 98.0, 99.5, 127.6–132.0, 169.1, 169.4, 169.7,
169.87, 169.90, 169.93, 170.3, 170.7; FABMS (pos.-ion): m/z 1008
[Mþ þ 1]; Anal. Calcd for C47H58NOS: C, 56.06; H, 5.81. found: C, 56.34;
H, 5.76.
ACKNOWLEDGMENTS
We thank Professors Makoto Kiso, Hideharu Ishida, and Robert
J. Linhardt for critical discussion and helpful suggestions.
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Received in Japan June 9, 2003

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