Organic & Biomolecular Chemistry
Paper
I
III–VII
II–VII
7
5 °C for 3 h and the solvent was reduced until 1/3 volume. 2.3 Hz, J5,6b = 6.6 Hz, H-5 ), 3.92–3.84 (m, 17H, H-3
, H-6
= 9.5 Hz, H-3 ), 3.87 (t, 1H, J = J3,4
2,3
),
=
II
The resulting residue was poured into acetone (150 mL), fil- 3.88 (t, 1H, J
3
,4
I
II–VII
tered and purified by column chromatography (6 : 3 : 1 MeCN– 9.3 Hz, H-3 ), 3.81–3.76 (m, 6H, H-5
NHCyst), 3.58 (2d, 6H, JH,H = 6.0 Hz, H-3Pent), 3.54 (dd, 1H,
MeCN–H O–NH OH); [α] = +115.3 (c 1.0 in DMSO); H NMR H-2 ), 3.54 (dd, 1H, J = 9.5 Hz, H-2 ), 3.56–5.49 (m, 11H,
), 3.73 (m, 4H, CH
2
N,
3
H
2
O–NH
4
OH) to give 3 in 67% yield (0.98 g); R
f
= 0.47 (6 : 3 : 1 CH
2
I
1
II
2
4
D
1,2
III–VII
II–VII
I
(500 MHz, 9 : 1 DMSO-d
6
–D
2
O, 333 K): δ 4.88–4.82 (m, 6H, H-2
, H-4
2
), 3.53 (m, 1H, H-4 ), 3.47 (m, 8H, CH NH,
), 4.84 (d, 1H, J1,2 = 3.0 Hz, H-1 ), 3.86–3.59 (m, 24H, H-1Pent), 3.18 (dd, 1H, J6a,6b = 12.8 Hz, H-6a ), 2.94 (dd, 1H,
II–VII
I
I
H-1
H-3
II–VII
II–VII
II–VII
I
3
, H-5
, H-6
), 3.82 (bt, 1H, J = J5,6a = 8.6 Hz, H-6b ), 2.88 (t, 2H, J
= 7.2 Hz, CH2SCyst), 2.87–2.78 (m, 6H,
H-5Pent), 2.17–1.98 (4 s, 36H, MeCO), 1.96 (m, 6H, H-4Pent);
C NMR (125.7 MHz, CD OD, 313 K): δ 182.0 (CS),
3
4
,5
H,H
I
I
II–VII
H-5 ), 3.65, (t, 1H, J2,3 = 8.6 Hz, H-3 ), 3.42–3.26 (m, 12H, H-2
H-4
,
II–VII
I
I
13
), 3.34 (dd, 1H, H-2 ), 3.26 (t, 1H, H-4 ), 3.08 (m, 2H,
I
I
I–VII
I
CH N ), 3.07 (bd, 1H, H-6a ), 2.71 (m, 1H, H-6b ), 2.64 (m, 170.9–170.1 (CO), 102.5–102.2 (C-1
), 84.6 (C-4 ), 82.6
2
Cyst
1
3
II–VII
I–VII
I–VII
2
H, CH
2
S
Cyst), 1.43 (bs, 9H, CMe
3
); C NMR (100.6 MHz, 9 : 1 (C-1Man), 81.8–81.7 (C-4
), 73.4–73.2 (C-3
), 72.9 (C-5
),
I–VII
I
I–VII
DMSO-d
6
–D O): δ 156.0 (CO), 102.9–101.2 (C-1
2
), 86.7 (C-4 ), 72.5–72.3 (C-2
), 71.1 (C-2Man), 69.8 (C-3Man), 69.5 (C-3Pent),
), 78.6 (C ), 73.5–72.4 (C-2 , C-3 , C-5 ), 69.1 (C-5Man), 66.3 (C-4Man), 62.4 (C-6Man), 60.8–60.6 (C-6
II–VII
I–VII
I–VII
I–VII
II–VII
82.3–82.0 (C-4
,
q
II–VII
I
I
6
0.6–60.2 (C-6
), 43.0 (CH
2
N
Cyst), 39.9 (CH
2
S
Cyst, C-6 ), 29.0 C-1Pent), 44.3 (CH
2
N, CH
2
N
Cyst), 33.3 (C-6 ), 32.1 (CH
2
S
Cyst),
(
CMe
3
); ESIMS: m/z 1316.9 [M + Na]+. Anal. Calcd for 29.3 (C-4Pent), 28.0 (C-5Pent), 19.4–19.1 (MeCO); ESIMS: m/z
2
+
C H NO S: C 45.47, H 6.46, N 1.08. Found: C 45.31, H 6.46, 1314.8 [M + 2 Na] . Anal. Calcd for C101H158N O S : C, 46.93;
4
9
83
36
2 64 5
N 0.96.
H, 6.16; N, 1.08; S, 6.20; found: C, 47.11; H, 6.34; N, 0.89;
S, 5.87.
6 -[2-[N′-[2,2,2-Tris[5-α-D-mannopyranosylthio)-2-oxapentyl]-
I
39
6
-(2-Aminoethylthio)cyclomaltoheptaose hydrochloride (4).
I
Compound 3 (94 mg, 72 μmol) was treated with a 1 : 1 TFA–
water mixture (3 mL) at rt for 1 h. The solvents were then evap- ethyl]-thioureido]ethylthio]cyclomaltoheptaose ((ManS)
3
-βCD).
orated under reduced pressure and acid traces were removed The trivalent mannosyl carrier (ManS) -βCD was obtained by
3
by co-evaporating with water several times. The residue was treating a solution of compound 6 (59 mg, 23 μmol) in dry
finally dissolved in diluted aq. HCl and freeze-dried to quanti- MeOH (3 mL) with methanolic NaOMe (1 m, 27 μL, 0.1 eq.) at
tatively furnish compound 4 as hydrochloride salt (89 mg). rt for 1 h. The reaction mixture was neutralized with Amberlite
1
+
[
(
5
3
J
α]D = +113.6 (c 0.9, H O); H NMR (500 MHz, D O): δ 5.04 IR-120 (H ) ion exchange resin, the resin filtered-off and the
2
2
II
I
d, 1H, J1,2 = 3.7 Hz, H-1 ), 4.99 (d, 1H, J1,2 = 3.7 Hz, H-1 ), solvent evaporated under reduced pressure to yield (ManS)
3
-
O)
III–VII
I
.00–4.97 (m, 5H, H-1
), 3.95 (bt, 1H, J4,5 = 9.4 Hz, H-5 ), βCD in 97% yield (46 mg). [α]
D
1
= +27.5 (c 1.0, H
2
O); UV (H
2
III–VII
III–VII
III–VII
.91–3.72 (m, 20H, H-3
, H-5
, H-6
), 3.86 (t, 1H, λmax = 262 nm (εmM = 2308); H NMR (500 MHz, D O, 323 K):
2
3
,4
II
I
II–VII
2,3 = J = 9.4 Hz, H-3 ), 3.85 (t, 1H, J2,3 = J3,4 = 9.4 Hz, H-3 ), δ 5.53 (bs, 3H, H-1Man), 5.33 (m, 6H, H-1
.78 (m, 2H, H-6 ), 3.77 (m, 1H, H-5 ), 3.61–3.44 (m, 12H,
), 3.58 (dd, 1H, H-2 ), 3.57 (m, 2H, H-2 , H-3
H-4 ), 3.47 (t, 1H, J4,5 = 9.4 Hz, H-4 ), 3.16 (t, 2H,
), 5.29 (d, 1H,
II
II
I
3
H-2
J1,2 = 3.0 Hz, H-1 ), 4.29 (bs, 3H, H-2Man), 4.19–3.99 (m, 31H,
III–VII
III–VII
II
I
II–VII
II–VII
II–VII
, H-4
, H-5
, H-6
, H-1Pent, H-3Man, H-4Man, H-5Man,
I
II
3
I
H,H
J =
H-6Man), 4.10 (t, 1H, J2,3 = J3,4 = 9.1 Hz, H-3 ), 4.09 (m, 1H,
I
II–VII
II–VII
6
.4 Hz, CH
2
N
Cyst), 3.07 (dd, 1H, J5,6a = 2.9 Hz, J6a,6b = 14.0 Hz, H-5 ), 3.97–3.85 (m, 12H, H-2
, H-4
), 3.93 (m, 2H,
I
I
I
3
H-6a ), 2.91 (dd, 1H, J
CH
8
C-5
= 6.5 Hz, H-6b ), 2.86 (t, 2H, CH N ), 3.90 (dd, 1H, H-2 ), 3.76 (t, 6H, JH,H = 5.4 Hz,
5
,6a
2 Cyst
1
3
I–VII
I
2
S
Cyst); C NMR (125.7 MHz, D
2
O): δ 101.9–101.7 (C-1
), 73.0–71.1 (C-2
), H-3Pent), 3.75 (t, 1H, H-4 ), 3.65 (s, 2H, CH
2
NH), 3.45 (bd, 1H,
I
II–VII
I–VII
I–VII
I
3
3.8 (C-4 ), 81.3–81.1 (C-4
, C-3
,
J
6a,6b = 11.2 Hz, H-6a ), 3.15 (t, 2H, JH,H = 6.5 Hz, CH SCyst),
2
I–VII
II–VII
I
I
), 60.5–60.2 (C-6
), 38.4 (C-6 ), 32.5 (CH NCyst), 29.7 3.07 (dd, 1H, J5,6b = 7.3 Hz, H-6b ), 2.99–2.87 (m, 6H, H-5Pent),
2
Cyst); ESIMS: m/z 1194.8 [M − Cl] .
+
13
(
CH
2
S
2
2.14 (m, 6H, H-4Pent); C NMR (125.7 MHz, D O, 323 K):
I
I–VII
I
6
-[2-[N′-[2,2,2-Tris[5-(2,3,4,6-tetra-O-acetyl-α-D-mannopyranosyl- δ 182.3 (CS), 102.6–102.1 (C-1
thio)-2-oxapentyl]ethyl]thioureido]ethylthio]cyclomaltoheptaose 81.3–80.9 (C-4
6). To a solution of 4 (94 mg, 72 μmol) and Et
44 μmol, 2 eq.) in dry DMF (1 mL), a solution of 5 in dry DMF 60.5–60.2 (C-6
), 85.3 (C-1Man), 85.0 (C-4 ),
II–VII
I–VII
), 73.6–71.4 (C-2Man, C-3Man, C-5Man, C-2
,
I–VII
I–VII
(
1
3
N (20 μL, C-3
, C-5
), 70.4 (C-3Pent), 67.2 (C-4Man), 61.5 (C-1Pent),
II–VII
I
2 2
), 44.7 (CH N, CH NCyst), 34.2 (C-6 ),
(
1 mL) was added. The reaction mixture was stirred at room 33.8 (CH2S ), 29.3 (C-4Pent), 28.0 (C-5Pent); ESIMS: m/z 1062.3
Cyst
2
+
3 134 2 52 5
temperature for 72 h preserving the pH at 8–9 with Et N. The [M + 2 Na] . Anal. Calcd for C77H N O S C, 44.46; H, 6.49;
reaction mixture was then concentrated and the residue was N, 1.35; S, 7.71; found: C, 44.25, H, 6.22; N, 1.03; S, 7.42.
purified by column chromatography (MeCN → 3 : 1 MeCN–
NMR titration experiments
H
H
2
2
O) to give 6 in 50% yield (93 mg). R
f
= 0.66 (6 : 3 : 1 MeCN–
1
O–NH OH); [α]
4
D
= +57.8 (c 1.0, MeOH); H NMR (500 MHz, Association constants (Kas) were determined in D
by measuring the proton chemical shift variations in the H
.6 Hz, J2,3 = 2.8 Hz, H-2Man), 5.30 (t, 3H, J3,4 = J4,5 = 9.8 Hz, NMR spectra of a solution of the βCD derivative (ManS) -βCD
H-4Man), 5.23 (dd, 3H, H-3Man), 5.04 (d, 1H, J1,2 = 3.7 Hz, H-1 ), in the presence of increasing amounts of the corresponding
2
O at 313 K
1
CD OD, 313 K): δ 5.39 (bs, 3H, H-1Man), 5.36 (d, 3H, J1,2
1
=
3
3
II
III–VII
I
5
4
.01–4.98 (m, 5H, H-1
), 5.00 (d, 1H, J1,2 = 3.3 Hz, H-1 ), iminosugar (6S-NOI-NJ or 6S-NAdB-NJ). The chemical shifts of
.41 (ddd, 3H, J5,6a = 4.9 Hz, J5,6b = 2.1 Hz, H-5Man), 4.30 the diagnostic signals obtained at 10–11 different CD:iminosu-
(dd, 3H, J6a,6b = 12.1 Hz, J5,6a = 5.2 Hz, H-6aMan), 4.16 (dd, 3H, gar concentration ratios were used in an iterative least-squares
J5,6b = 5.2 Hz, H-6bMan), 3.97 (ddd, 1H, J4,5 = 9.3 Hz, J5,6a
=
fitting procedure. For a detailed description, see ESI.†
This journal is © The Royal Society of Chemistry 2014
Org. Biomol. Chem., 2014, 12, 2289–2301 | 2297