Alkylating agents from sugars. Cyclophosphamides derived from 2-amino- 2-deoxy-D-allose

Article abstract of DOI:10.1016/S0008-6215(98)00056-1

Cyclophosphamides derived from alkyl 2-amino-4,6-O-benzylidene-2-deoxy- β-D-allopyranosides have been synthesized with good yield by treatment of the corresponding 2-amino-2-deoxy-D-allose derivatives with bis(2- chloroethyl)phosphoramide dichloride. The ring-forming reaction took place with very high diastereoselectivity. Subsequent hydrogenolysis gave excellent yields of cyclophosphamides derived from alkyl 2-amino-2-deoxy-β-D- allopyranosides, with hydrophilicity greater than that of the precursors. The starting material was easily available from 2-acetamido-2-deoxy-D-glucose.

Full text of DOI:10.1016/S0008-6215(98)00056-1

Carbohydrate Research 308 (1998) 57±62
Alkylating agents from sugars.
Cyclophosphamides derived from
2-amino-2-deoxy-d-allose
Fernando Iglesias-Guerra *, Isidora Romero, Felipe Alcudia,
Jose M. Vega-Perez *
Â
Departamento de Quimica Organica y Farmaceutica, Facultad de Farmacia, Universidad de Sevilla, E-
Â
Â
41071 Sevilla, Spain
Received 30 October 1997; accepted 22 February 1998
Abstract
Cyclophosphamides derived from alkyl 2-amino-4,6-O-benzylidene-2-deoxy-ꢀ-d-allopyrano-
sides have been synthesized with good yield by treatment of the corresponding 2-amino-2-deoxy-
d-allose derivatives with bis(2-chloroethyl)phosphoramide dichloride. The ring-forming reaction
took place with very high diastereoselectivity. Subsequent hydrogenolysis gave excellent yields of
cyclophosphamides derived from alkyl 2-amino-2-deoxy-ꢀ-d-allopyranosides, with hydrophilicity
greater than that of the precursors. The starting material was easily available from 2-acetamido-2-
deoxy-d-glucose. # 1998 Elsevier Science Ltd. All rights reserved
Keywords: Alkylating agents; Cyclophosphamides from sugars ; 2-amino-2-deoxy-d-allose derivatives
1. Introduction
selective carriers, since the cancerous cellsÐunder-
going rapid growthÐhave a greater demand for
them than do healthy cells.
In the present report, the synthesis of cyclopho-
sphamides derived from sugars, of which there is
scarce mention in the scienti®c literature [2±4] is
presented. Moreover, the described compounds
contain di€erent lipophilic moieties in order to
modulate the hydrophilic-lipophilic balance (HLB)
of these compounds.
Alkylating agents have been pioneering drugs in
the treatment of malignant metastases. Among
them, cyclophosphamide, still has a broad spec-
trum of action against a great variety of neoplasias,
and is commonly used to this end, both alone and
in combined treatments [1]. Despite the advantages
it o€ers, it is not free of drawbacks, partly because
of the possible inducement of resistance, and also
because of its low selectivity. The synthesis of new,
potentially more selective drugs could help to solve
these two problems.
2. Results and discussion
One way of improving the targeting of the drug
is to couple it with primary metabolites that act as
The synthesis of cyclophosphamides derived
from 2-acetamido-2-deoxy-d-allose (17±22) described
in this paper was carried out using commercial
* Corresponding authors.
0008-6215/98/$19.00 # 1998 Elsevier Science Ltd. All rights reserved
PII S0008-6215(98)00056-1

58
F. Iglesias-Guerra et al./Carbohydrate Research 308 (1998) 57±62
2-acetamido-2-deoxy-d-glucose as starting sugar.
The latter was transformed into the corresponding
alkyl 2-amino-4,6-O-benzylidene-2-deoxy-ꢀ-d-allopy-
ranosides (14±16) by a sequence of reactions
described in the literature for the corresponding
benzyl glycoside [5±7] (Scheme 1).
1
The most characteristic H NMR signals of 14±
16 were those corresponding to H-1 at 4.6±4.5 ppm
as a doublet (J1,2 8.1 Hz), H-2 at 2.7 ppm as a
double doublet (J1,2 8.1, J2,3 2.6 Hz) and H-3 at
4.2 ppm as a triplet (J2,3=J3,4 2.6 Hz). These
assignments were con®rmed by double resonance
experiments on 16.
Scheme 2.
The cyclophosphamides 17±19 were obtained by
reaction of 14±16 with bis(2-chloroethyl)pho-
sphoramide dichloride in dichloromethane at 0 C
ꢀ
in the 1H NMR spectra, and, above all, because the
31P NMR spectra presented a single signal for each
of compounds 17±19, at ꢁ 25.96, 26.10 and
26.08 ppm, respectively.
under an inert atmosphere (Scheme 2). Com-
pounds 17±19 were puri®ed on a silica gel column,
and obtained as perfectly stable, white solids in
yields of 78±87%. Owing to the cis fusion of the
®ve-membered ring to the pyranose ring, these
molecules are free of angular strain. The NMR
Compounds 17±19 were very soluble in dichlor-
omethane, moderately soluble in methanol and
ethanol, and totally insoluble in water. In order to
modulate the HLB of these substances and increase
their hydrosolubility, we prepared the cyclopho-
sphamides 20±22, obtained readily in excellent
yield (>85%) by hydrogenolysis in the presence of
1
data con®rmed the proposed structures. In the H
NMR spectrum of 19, for example, the signals for
H-1 (a doublet with J1,2 8.1 Hz at 4.86 ppm), H-2
(part of the multiplet at 4.1±3.2 ppm), and H-3 (a
triplet with J2,3=J3,4 3.5 Hz at 4.73 ppm) were
observed at low-®eld as compared with the spec-
trum of the precursor 16.
It is also noteworthy that the ring-forming reac-
tion took place with high diastereoselectivity with
regard to the substituents on the chiral phosphorus
atom, as shown by the absence of signal duplicity
1
palladium catalyst. The H NMR spectrum of 21,
for example, showed the signals for OH-4 at
5.16 ppm as a doublet (J4,OH 7.1 Hz) and for OH-6
at 4.54 ppm as a triplet (J6,OH 5.8 Hz). The 31P
NMR spectra indicated a single diastereomer.
These compounds showed moderate solubility in
both dichloromethane and water, varying gradu-
ally depending on the nature of the glycoside
moiety. COSY and CHCORR experiments for 21
con®rmed the predicted structure of these cyclo-
phosphamides.
The cyclophosphamides synthesized will be tes-
ted for cytotoxicity and inhibition of cell growth
(cytostatic activity) using the appropriate cell lines
in in vitro experiments.
3. Experimental
General methods.ÐEvaporations were con-
ducted under reduced pressure. Preparative chro-
matography was performed on Silica Gel 60 (E.
Merck). Kieselgel 60 F254 (E. Merck) was used for
TLC. Melting points are uncorrected. Optical
rotations were obtained on a Bellingham+Stanley
ꢀ
Scheme 1.
Ltd P-20 polarimeter at 25 C. Mass spectra were

F. Iglesias-Guerra et al./Carbohydrate Research 308 (1998) 57±62
59
recorded on a Kratos MS-80-RFA mass spectro-
meter at 70 eV for EI and 150 eV for CI. FAB mass
spectra were recorded using a thioglycerol matrix.
J2,NH 8.6 Hz, N±H), 5.29 (dd, 1-H, J2,3 10.5, J3,4
9.3 Hz, H-3), 5.04 (t, 1-H, J3,4=J4,5 9.5 Hz, H-4),
4.66 (d, 1-H, J1,2 8.3 Hz, H-1), 4.25 (dd, 1-H, J5,6
ꢀ
0
0
0
NMR spectra were recorded at 25 C on a Bruker
1
4.7, J6,6 12.2 Hz, H-6), 4.10 (dd, 1-H, J5,6 2.6, J6,6
AC-200 spectrometer at 200 MHz for H, 50 MHz
for 13C, and 81 MHz for 31P, and on a Bruker
12.2 Hz, H-60), 3.9±3.6 (m, 3-H, H-2, H-5,
OCHHR), 3.45 (m, 1-H, OCHHR), 2.06, 2.01,
2.00, 1.92 (4-s, 4CH3), 1.7±1.1 [(CH2)6], 0.85 (t, 3-
H, J 6.4 Hz, CH3); 13C NMR (50 MHz, CDCl3): ꢁ
170.9, 170.8, 170.1, 169.4 (4C=O), 100.7 (C-1),
72.3 (C-4), 71.8 (C-3), 70.0 (OCH2R), 68.7 (C-5),
62.2 (C-6), 55.0 (C-2), 31.8, 29.4, 29.3, 29.2, 25.9,
22.6 [(CH2)6], 23.3 (CH3CON), 20.8, 20.7, 20.6 (4-
CH3COO), 14.1 (CH3). Anal. Calcd for C22H37
NO9: C, 57.50; H, 8.12; N, 3.05. Found: C, 57.54;
H, 8.18; N, 3.00.
1
AMX-500 spectrometer at 500 MHz for H and
125 MHz for 13C. The chemical shifts are reported
in ppm on the ꢁ scale relative to Me4Si and H3PO4.
Compounds 4, 7, 10 and 13 were synthesized in
collaboration with other colleagues in relation with
other works and will be published elsewhere [8];
their elemental analyses and physical data are in
agreement with the proposed structures.
Alkyl 2-acetamido-3,4,6-tri-O-acetyl-2-deoxy-b-
d-glucopyranosides (2±4).ÐA soln of 2-acetamido-
3,4,6-tri-O-acetyl-2-deoxy-ꢂ-d-glucopyranosyl
chloride [9] (1) (18.3 g, 50 mmol) in dry MeNO2
(40 mL) was added dropwise, with exclusion of
moisture, to a stirred mixture of alcohol (cyclo-
hexanol, 1-octanol, 1-dodecanol; 30 mL in each
case) in anhyd toluene (40 mL) containing 4 A
molecular sieves (30 g) and Hg(CN) (12.5 g). The
mixture was stirred overnight at room temperature.
When TLC showed that all of the chloride had
reacted, the mixture was diluted with EtOAc and
®ltered through a pad (12 cm diameterÂ1 cm
height) of alumina. The organic layer was washed
with an aq satd soln of NaHCO3 and H2O, then
dried (Na2SO4) and concentrated. The residue was
suspended in hexane with stirring for 1 h. The solid
obtained was ®ltered and washed with hexane.
Cyclohexyl glycoside (2).ÐRecrystallized from
Alkyl 2-acetamido-4,6-O-benzylidene-2-deoxy-b-
d-glucopyranosides (5±7).ÐTo a soln of alkyl 2-
acetamido-3,4,6-tri-O-acetyl-2-deoxy-ꢀ-d-glucopy-
ranoside (2±4) (20 mmol) in MeOH (200 mL) was
added a soln of NaOMe (4 mmol) in MeOH
(20 mL). After 30 min at room temperature, the
mixture was neutralized by addition of Dowex 50
resin (H+ form), ®ltered and evaporated. The solid
obtained was dissolved in benzaldehyde (100 mL),
and ZnCl2 (5 g) was added. The mixture was stirred
overnight and then poured into 1:1 hexane±H2O
(500 mL) with stirring. The precipitate was ®ltered,
washed with H2O and with hexane, and then
recrystallized from EtOH.
Cyclohexyl derivative (5).Ð6.0 g (77%); mp 259±
261 ꢀC; [ꢂ]d 122.4ꢀ (c 0.7, DMF); CIMS: m/z 392
1
(31%) [MH+]; H NMR (200 MHz, Me2SO-d6): ꢁ
7.80 (d, 1-H, J2,NH 8.9 Hz, N±H), 7.5±7.3 (m, 5-H,
Ph), 5.56 (s, 1-H, PhCH), 5.25 (d, 1-H, J3,OH
5.5 Hz, O±H), 4.56 (d, 1-H, J1,2 8.3 Hz, H-1), 4.16
(dd, 1-H, J5,6e 4.5, J6e,6a 10.1 Hz, H-6e), 3.8±3.2 (m,
6-H, H-2, H-3, H-4, H-5, H-6a, OCH), 1.80 (s,
CH3), 2.0±1.0 [(CH2)5]; 13C NMR (50 MHz,
Me2SO-d6): ꢁ 169.4 (C=O), 137.9, 129.0, 128.2,
126.5 (Ph), 100.7 (PhCH), 100.1 (C-1), 81.4 (C-4),
76.1 (OCH), 70.5 (C-3), 68.1 (C-6), 66.0 (C-5), 56.7
(C-2), 33.0, 31.1, 25.3, 23.2, 22.9 [(CH2)5], 23.1
(CH3). Anal. Calcd for C21H29NO6: C, 64.43; H,
7.47; N, 3.58. Found: C, 64.22; H, 7.39; N, 3.57.
1-Octyl derivative (6).Ð6.4 g (76%); mp 191±
ꢀ
EtOH; 17.8 g (83%); mp 176±178 C; [ꢂ]d 37.5ꢀ
1
(c 0.8, CH2Cl2); CIMS: m/z 430 (25%) [MH+]; H
NMR (200 MHz, CDCl3): ꢁ 6.00 (d, 1-H, J2,NH
8.5 Hz, N±H), 5.34 (dd, 1-H, J2,3 10.5, J3,4 9.5 Hz,
H-3), 4.97 (t, 1-H, J3,4=J4,5 9.5 Hz, H-4), 4.81 (d,
0
1-H, J1,2 8.3 Hz, H-1), 4.21 (dd, 1-H, J5,6 5.0, J6,6
0
0
12.1 Hz, H-6), 4.04 (dd, 1-H, J5,6 2.5, J6,6 12.1 Hz,
H-60), 3.6 (m, 3. H, H-2, H-5, OCH), 2.00, 1.96,
1.95, 1.87 (4s, 4 CH3), 2.0±1.0 ((CH2)5); 13C NMR
(50 MHz, CDCl3): ꢁ 170.6, 170.5, 170.3, 169.4 (4
C=O), 98.8 (C-1), 77.6 (OCH), 72.1 (C-4), 71.3 (C-
3), 68.9 (C-5), 62.3 (C-6), 55.2 (C-2), 33.1, 31.6,
25.4, 23.7, 23.6 [(CH2)5], 23.2, 20.6, 20.5 (4 CH3).
Anal. Calcd for C20H31NO9: C, 55.93; H, 7.28; N,
3.26. Found: C, 55.87; H, 7.04; N, 3.52.
ꢀ
193 C; [ꢂ]d 84.7ꢀ (c 1.0, CH2Cl2); CIMS: m/z
422 (45%) [MH+]; H NMR (200 MHz, Me2SO-
1
d6): ꢁ 7.81 (d, 1-H, J2,NH 8.6 Hz, N±H), 7.5±7.3 (m,
5-H, Ph), 5.58 (s, 1-H, PhCH), 5.24 (d, 1-H, J3,OH
5.2 Hz, O±H), 4.44 (d, 1-H, J1,2 8.0 Hz, H-1), 4.18
(dd, 1-H, J5,6e 4.2, J6e,6a 9.9 Hz, H-6e), 3.8±3.2 (m,
7-H, H-2, H-3, H-4, H-5, H-6a, OCH2R), 1.79 (s,
1-Octyl glycoside (3).ÐRecrystallized from
Et2O±hexane.Ð18.4 g (80%); mp 126±127 ꢀC;
[ꢂ]d 36.4ꢀ (c 0.6, CH2Cl2); CIMS: m/z 460 (48%)
[MH+]; 1H NMR (200MHz, CDCl3): ꢁ 5.42 (d, 1-H,

60
F. Iglesias-Guerra et al./Carbohydrate Research 308 (1998) 57±62
3-H, CH3CON), 1.5±1.1 [(CH2)6], 0.85 (t, 3-H, J
6.4 Hz, CH3); 13C NMR (50 MHz, Me2SO-d6): ꢁ
168.9 (C=O), 137.7, 128.8, 128.0, 126.3 (Ph), 101.6
(PhCH), 100.6 (C-1), 81.3 (C-4), 70.4 (C-3), 68.7
(OCH2R), 67.9 (C-6), 65.9 (C-5), 56.2 (C-2), 31.2,
29.0, 28.7, 25.3, 22.1 [(CH2)6], 23.0 (CH3CON),
13.9 (CH3). Anal. Calcd for C23H35NO6: C, 65.54;
H, 8.37; N, 3.32. Found: C, 65.49; H, 8.53; N, 3.30.
Alkyl 2-acetamido-4,6-O-benzylidene-2-deoxy-3-
O-methylsulfonyl-b-d-glucopyranosides (8±10).Ð
Alkyl 2-acetamido-4,6-O-benzylidene-2-deoxy-ꢀ-d-
glucopyranoside (5±7) (15 mmol) was dissolved in
pyridine (50 mL), aꢀnd MeSO2Cl (3 mL) was added
with cooling to 0 C. The mixture was stored at
amido-4,6-O-benzylidene-2-deoxy-3-O-methylsul-
fonyl-ꢀ-d-glucopyranoside (8±10) (7.5 mmol) and
anhyd NaAcO (3 g) in 96:4 2-methoxyethanol±H2O
(50 mL) was heated at the re¯ux temperature for
12 h. After cooling, the mixture was poured into
H2O, and the precipitate was collected by ®ltration
and recrystallized from EtOH.
Cyclohꢀexyl derivative (11).Ð2.1 g (72%); mp
214±216 C; [ꢂ]d 209.8ꢀ (c 0.6, CH2Cl2); CIMS:
m/z 392 (23%) [MH+]; 1H NMR (200 MHz,
Me2SO-d6): ꢁ 7.85 (d, 1-H, J2,NH 9.1 Hz, N±H),
7.5±7.3 (m, 5-H, Ph), 5.62 (s, 1-H, PhCH), 5.42 (d,
1-H, J3,OH 3.7 Hz, O±H), 4.69 (d, 1-H, J1,2 8.6 Hz,
H-1), 4.21 (dd, 1-H, J5,6e 4.8, J6e,6a 9.6 Hz, H-6e),
3.91 (m, 1-H, H-3), 3.85±3.45 (m, 5-H, H-2, H-4,
H-5, H-6a, OCH), 1.82 (s, CH3), 1.8±1.1 [(CH2)5];
13C NMR (50 MHz, Me2SO-d6): ꢁ 168.6 (C=O),
137.8, 128.8, 127.9, 126.4 (Ph), 100.6 (PhCH), 98.2 (C-
1), 78.6 (C-4), 76.2 (OCH), 68.4 (C-6), 67.5 (C-3), 62.6
(C-5), 52.8 (C-2), 33.1, 31.2, 25.2, 23.3, 22.6 [(CH2)5],
23.0 (CH3). Anal. Calcd for C21H29NO6: C, 64.43; H,
7.47; N, 3.58. Found: C, 64.11; H, 7.33; N, 3.52.
1-Octyl derivative (12).Ð2.5 g (78%); mp 221±
ꢀ
0 C for 24 h and poured into ice-water. The solid
was collected by ®ltration, washed with H2O, dried
(Na2SO4), and was recrystallized from EtOH.
Cyclohexyl derivative (8).Ð5.3 g (76%); mp 170±
ꢀ
172 C; [ꢂ]d 45.2ꢀ (c 1.7, DMF); FABMS: m/z
492 (100%) [MNa+]; 1H NMR (200 MHz,
CDCl3): ꢁ 7.5±7.3 (m, 5H, Ph), 5.80 (d, 1-H, J2,NH
7.2 Hz, N-H), 5.50 (s, 1-H, PhCH), 5.33 (m, 2-H,
H-1, H-3), 4.36 (dd, 1-H, J5,6e 4.7, J6e,6a 10.3 Hz,
H-6e), 3.9±3.5 (m, 4-H, H-4, H-5, H-6a, OCH), 3.24
(m, 1-H, H-2), 2.90 (s, 3-H, CH3SO3), 1.99 (s,
CH3CON), 1.95±1.15 [m, 10-H, (CH2)5]; 13C NMR
(50 MHz, CDCl3): ꢁ 171.3 (C=O), 136.6, 129.4,
128.4, 126.0 (Ph), 100.8 (PhCH), 98.3 (C-1), 79.2
(C-3), 78.4 (C-4), 78.2 (OCH), 68.7 (C-6), 65.4 (C-
5), 58.0 (C-2), 38.5 (CH3SO3), 33.3, 31.7, 25.4,
23.9, 23.5 [(CH2)5], 23.7 (CH3CON). Anal. Calcd
for C22H31NO8S: C, 56.27; H, 6.67; N, 2.98.
Found: C, 56.26; H, 6.63; N, 2.89.
ꢀ
223 C; [ꢂ]d 97.6ꢀ (c 2.1, CH2Cl2); CIMS: m/z
422 (22%) [MH+]; H NMR (200 MHz, Me2SO-
1
d6): ꢁ 7.92 (d, 1-H, J2,NH 8.8 Hz, N±H), 7.5±7.3 (m,
5-H, Ph), 5.62 (s, 1-H, PhCH), 5.43 (d, 1-H, J3,OH
4.3 Hz, O±H), 4.61 (d, 1-H, J1,2 8.7 Hz, H-1), 4.21
(dd, 1-H, J5,6e 4.6, J6e,6a 9.5 Hz, H-6e), 3.94 (m, 1-
H, H-3), 3.8±3.4 (m, 6-H, H-2, H-4, H-5, H-6a,
OCH2R), 1.82 (s, 3H, CH3CON), 1.5±1.2 [(CH2)6],
0.85 (t, 3-H, J 6.4 Hz, CH3); 13C NMR (50 MHz,
Me2SO-d6): ꢁ 168.6 (C=O), 137.8, 128.6, 127.9,
126.4 (Ph), 100.6 (PhCH), 99.5 (C-1), 78.6 (C-4),
68.8 (OCH2R), 68.3 (C-6), 67.4 (C-3), 62.7 (C-5),
52.7 (C-2), 31.2, 29.1, 28.7, 25.4, 22.1 [(CH2)6], 22.6
(CH3CON), 13.9 (CH3). Anal. Calcd for
C23H35NO6: C, 65.54; H, 8.37; N, 3.32. Found: C,
65.57; H, 8.21; N, 3.26.
1-Octyl derivative (9).Ð5.3 g (71%); mp 192±
ꢀ
194 C; [ꢂ]d 375.9ꢀ (c 1.0, CH2Cl2); CIMS: m/z
1
500 (5%) (MH+); H NMR (200 MHz, CDCl3): ꢁ
7.5±7.3 (m, 5H, Ph), 5.89 (d, 1-H, J2,NH 7.7 Hz, N±
H), 5.50 (s, 1-H, PhCH), 5.33 (t, 1-H, J2,3=J3,4
10.0 Hz, H-3), 5.09 (d, 1-H, J1,2 8.2 Hz, H-1), 4.36
(dd, 1-H, J5,6e 4.6, J6e,6a 10.2 Hz, H-6e), 3.9±3.3 (m,
6-H, H-2, H-4, H-5, H-6a, OCH2R), 2.91 (s, 3-H,
CH3SO3), 2.00 (s, 3-H, CH3CON), 1.7±1.1 [m,
12H, (CH2)6], 0.86 (t, 3-H, J 6.4 Hz, CH3); 13C
NMR (50 MHz, CDCl3): ꢁ 171.3 (C=O), 136.6,
129.4, 128.4, 126.0 (Ph), 101.7 (PhCH), 100.3 (C-
1), 79.2 (C-3), 78.6 (C-4), 70.6 (OCH2R), 68.7 (C-
6), 65.5 (C-5), 57.2 (C-2), 38.5 (CH3SO3), 31.8,
29.5, 29.3, 25.8, 22.6 [(CH2)6], 23.4 (CH3CON), 14.1
(CH3). Anal. Calcd for C24H37NO8S: C, 57.71; H,
7.41; N, 2.80. Found: C, 57.53; H, 7.20; N, 2.79.
Alkyl 2-acetamido-4,6-O-benzylidene-2-deoxy-b-
d-allopyranosides (11±13).ÐA soln of alkyl 2-acet-
Alkyl
2-amino-4,6-O-benzylidene-2-deoxy-b-d-
allopyranosides (14±16).ÐA soln of alkyl 2-
acetamido-4,6-O-benzylidene-2-deoxy-ꢀ-d-allopy-
ranoside (11±13) (8 mmol) in a hot mixture of
KOH (15 g) and 96% EtOH (50 mL) was heated
under re¯ux for 5±6 h. The hot soln was poured
carefully into hot H2O (400 mL). After being
cooled to room temperꢀature, the precipitate was
kept overnight at 10 C. The solid was ®ltered
o€, washed with H2O, dried, and then crystallized
from EtOH.
Cyclohꢀexyl derivative (14).Ð2.6 g (94%); mp
186±188 C; [ꢂ]d 219.1ꢀ (c 1.6, CH2Cl2); CIMS:

F. Iglesias-Guerra et al./Carbohydrate Research 308 (1998) 57±62
61
m/z 350 (13%) [MH+]; 1H NMR (200 MHz,
CDCl3): ꢁ 7.5±7.3 (m, 5H, Ph), 5.54 (s, 1-H,
PhCH), 4.62 (d, 1-H, J1,2 8.1 Hz, H-1), 4.33 (dd, 1-
H, J5,6e 4.8, J6e,6a 10.0 Hz, H-6e), 4.23 (t, 1-H,
J2,3=J3,4 2.6 Hz, H-3), 3.96 (td, 1-H, J4,5=J5,6a
9.8, J5,6e 4.8 Hz, H-5), 3.74 (t, 1-H, J5,6a=J6e,6a
10.1 Hz, H-6a), 3.6 (m, 2-H, H-4, OCH), 2.68 (dd,
mide dichloride (5.5 mmol) was added quickly. The
reaction mixture was stirred overnight at room
temperature. At the end of the reaction (TLC) the
mixture was diluted with CH2Cl2, and washed
successively, with aq 1 N HCl, aq satd soln of
NaHCO3, and H2O, then dried (Na2SO4) and con-
centrated to give a solid product. The product
obtained was puri®ed by ¯ash chromatography on
silica gel using CH2Cl2±2-PrOH (120:1) as eluent.
A small fraction of pure compound was recrys-
tallized from EtOH.
1-H, J1,2 8.1, J2,3 2.8 Hz, H-2), 2.0-1.1 [(CH2)5]; 13
C
NMR (50 MHz, CDCl3): ꢁ 137.2, 129.2, 128.3,
126.2 (Ph), 101.8 (C-1), 101.0 (PhCH), 79.6 (C-4),
77.5 (OCH), 69.3 (C-6), 69.1 (C-3), 63.5 (C-5), 55.1
(C-2), 33.7, 31.9, 25.5, 24.2, 24.0 [(CH2)5]. Anal.
Calcd for C19H27NO5: C, 65.33; H, 7.73; N, 4.01.
Found: C, 65.05; H, 7.60; N, 4.05.
Cyclohꢀexyl derivative (17).Ð2.35 g (87%); mp
220±222 C; [ꢂ]d 46.2ꢀ (c 0.8, MeOH); FABMS:
1
m/z 557 (100%) [MNa+]; H NMR (500 MHz,
1-Octyl derivative (15).Ð2.7 g (88%); mp 153±
ꢀ
CDCl3): ꢁ 7.6±7.3 (m, 5-H, Ph), 5.56 (s, 1-H,
PhCH), 5.04 (d, 1-H, J1,2 7.6 Hz, H-1), 4.78 (t, 1-H,
J2,3=J3,4 3.4 Hz, H-3), 4.44 (dd, 1-H, J5,6e 5.3,
J6e,6a 10.5 Hz, H-6e), 4.01 (td, 1-H, J4,5=J5,6a 9.9,
J5,6e 5.6 Hz, H-5), 3.8±3.1 [m, 13-H, H-2, H-4, H-
6a, N-H, OCH, N(CH2CH2Cl)2], 2.1±1.1 [(CH2)5];
13C NMR (125 MHz, CDCl3): ꢁ 137.2, 129.3,
128.3, 126.4 (Ph), 102.7 (C-1), 101.6 (PhCH), 78.0,
77.0, 76.7 (OCH, C-3, C-4), 68.9 (C-6), 63.1 (C-5),
55.2 (C-2), 49.2 (NCH2), 42.0 (CH2Cl), 33.6, 31.9,
25.4, 24.2, 24.0 [(CH2)5]. 31P NMR (81 MHz,
CDCl3): ꢁ 25.96. Anal. Calcd for C23H33Cl2N2O6P:
C, 51.59; H, 6.21; N, 5.23. Found: C, 51.64; H,
6.12; N, 5.13.
155 C; [ꢂ]d 59.2ꢀ (c 1.0, CH2Cl2); CIMS: m/z
1
380 (9%) [MH+]; H NMR (200 MHz, CDCl3): ꢁ
7.5±7.3 (m, 5H, Ph), 5.53 (s, 1-H, PhCH), 4.49 (d,
1-H, J1,2 8.1 Hz, H-1), 4.34 (dd, 1-H, J5,6e 4.7, J6e,6a
10.0 Hz, H-6e), 4.22 (m, 1-H, H-3), 4.0±3.3 (m, 5-H,
H-4, H-5, H-6a, OCH2R), 2.69 (dd, 1-H, J1,2 8.1,
J2,3 2.5 Hz, H-2), 1.7±1.1 [(CH2)6], 0.86 (t, 3H, J
6.4 Hz, CH3); 13C NMR (50 MHz, CDCl3): ꢁ 137.2,
129.2, 128.3, 126.2 (Ph), 102.9 (C-1), 101.9
(PhCH), 79.7 (C-4), 70.4 (OCH2R), 69.2 (C-6), 68.9
(C-3), 63.4 (C-5), 55.2 (C-2), 31.8, 29.6, 29.4, 29.2,
26.0, 22.6 [(CH2)6], 14.1 (CH3). Anal. Calcd for
C21H33NO5: C, 66.46; H, 8.76; N, 3.69. Found: C,
66.49; H, 8.55; N, 3.60.
1-ꢀOctyl derivative (18).Ð2.40 g (85%); mp 122±
124 C; [ꢂ]d 51.0ꢀ (c 1.0, CH2Cl2); FABMS: m/z
587 (100%) [MNa+]; 1H NMR (500 MHz, Me2SO-
d6): ꢁ 7.5±7.3 (m, 5-H, Ph), 5.76 (dd, 1-H, J2,NH 2.5,
2JH,P 17.6 Hz, N±H), 5.66 (s, 1-H, PhCH), 4.71 (dd,
1-H, J2,3 3.5, J3,4 5.0 Hz, H-3), 4.58 (d, 1-H, J1,2
6.9 Hz, H-1), 4.29 (dd, 1-H, J5,6e 5.6, J6e,6a 9.8 Hz,
H-6e), 3.9±3.3 [m, 14-H, H-2, H-4, H-5, H-6a,
OCH2R, N(CH2CH2Cl)2], 1.5±1.2 [(CH2)6], 0.85 (t,
3H, J 6.9 Hz, CH3); 13C NMR (50 MHz, CDCl3): ꢁ
136.8, 129.3, 128.3, 126.4 (Ph), 103.6 (C-1), 102.7
(PhCH), 77.2, 76.7 (C-3, C-4), 70.7 (OCH2R), 68.9
(C-6), 62.8 (C-5), 57.2 (C-2), 49.2 (NCH2), 42.0
(CH2Cl), 31.8, 29.6, 29.3, 29.2, 25.9, 22.6 [(CH2)6].
31P NMR (81 MHz, CDCl3): ꢁ 26.10. Anal. Calcd
for C25H39Cl2N2O6P: C, 53.10; H, 6.95; N, 4.95.
Found: C, 53.13; H, 6.86; N, 4.80.
1-Dodecyl derivative (16).Ð2.6 g (75%); mp 117±
119 ꢀC; [ꢂ]d 103.7ꢀ (c 0.5, CH2Cl2); FABMS: m/z
458 (100%) [MNa+]; 1H NMR (200 MHz,
CDCl3): ꢁ 7.5±7.3 (m, 5-H, Ph), 5.54 (s, 1-H,
PhCH), 4.50 (d, 1-H, J1,2 8.1 Hz, H-1), 4.34 (dd, 1-
H, J5,6e 4.7, J6e,6a 10.1 Hz, H-6e), 4.22 (t, 1-H,
J2,3=J3,4 2.5 Hz, H-3), 4.0±3.4 (m, 5-H, H-4, H-5,
H-6a, OCH2R), (2.69 dd, 1-H, J1,2 8.1, J2,3 2.6 Hz,
H-2), 1.7±1.1 [(CH2)10], 0.84 (t, 3-H, J 6.6 Hz,
CH3). 13C NMR (50 MHz, CDCl3): ꢁ 137.2, 129.2,
128.3, 126.2 (Ph), 102.9 (C-1), 101.8 (PhCH), 79.7
(C-4), 70.4 (OCH2R), 69.2 (C-6), 69.0 (C-3), 63.4
(C-5), 55.2 (C-2), 31.9, 29.6, 29.5, 29.4, 29.3, 26.0,
22.7 [(CH2)10], 14.1 (CH3). Anal. Calcd for
C25H41NO5: C, 68.93; H, 9.49; N, 3.22. Found: C,
69.00; H, 9.28; N, 3.11.
Alkyl 2-amino-4,6-O-benzylidene-2-N-3-O-[bis (2-
chloroethyl)amino]phosphoryl-2-deoxy-b-d-allopyra
nosides (17±19).ÐA soln of alkyl 2-amino-4,6-O-
1-Dodecyl derivative (19).Ð2.45 g (78%); mp
ꢀ
133±135 C; [ꢂ]d 40.7ꢀ (c 0.9, CH2Cl2); FABMS:
1
m/z 643 (100%) [MNa+]; H NMR (200 MHz,
benzylidene-2-deoxy-ꢀ-d-allopyranoside
(5 mmol) and Et3N (20 mL) in distilled and dried
(14±16)
CDCl3): ꢁ 7.6±7.3 (m, 5-H, Ph), 5.54 (s, 1-H,
PhCH), 4.86 (d, 1-H, J1,2 8.1 Hz, H-1), 4.73 (t, 1-H,
J2,3=J3,4 3.5 Hz, H-3), 4.39 (dd, 1-H, J5,6e 4.8,
J6e,6a 10.2 Hz, H-6e), 4.1±3.2 [m, 15-H, H-2, H-4,
CH2Cl2 (200 mL) under argon atmosphere was
ꢀ
cooled to 0 C. Then bis(2-chloroethyl)phosphora-

62
F. Iglesias-Guerra et al./Carbohydrate Research 308 (1998) 57±62
H-5, H-6a, N-H, OCH2R, N(CH2CH2Cl)2], 1.7±1.1
[(CH2)10], 0.86 (t, 3-H, J 6.6 Hz, CH3); 13C NMR
(50 MHz, CDCl3): ꢁ 136.8, 129.4, 128.4, 126.4 (Ph),
103.6 (C-1), 102.7 (PhCH), 77.1, 76.7 (C-3, C-4),
70.7 (OCH2R), 68.9 (C-6), 62.8 (C-5), 57.1 (C-2),
49.3, 49.2 (NCH2), 42.1 (CH2Cl), 31.9, 29.6, 29.3,
25.9, 22.7 [(CH2)10]. 31P NMR (81 MHz, CDCl3): ꢁ
26.08. Anal. Calcd for C29H47Cl2N2O6P: C, 56.04;
H, 7.62; N, 4.51. Found: C, 56.15; H, 7.51; N, 4.30.
Alkyl 2-amino-2-N-3-O-[bis(2-chloroethyl) amino]
phosphoryl-2-deoxy-b-d-allopyranosides (20±22).Ð
A soln of alkyl 2-amino-4,6-O-benzylidene-2-N-3-
O-[bis(2-chloroethyl)amino]phosphoryl-2-deoxy-ꢀ-
d-allopyranoside (17±19) (0.3 mmol) in MeOH
(30 mL) was hydrogenolyzed over 10% Pd(C)
(30 mg) at room temperature and a pressure of 50
PSI in a Parr hydrogenation apparatus. TLC indi-
cated complete reaction after 12±20 h. The mixture
was diluted with MeOH, the catalyst was ®ltered
o€ and washed with MeOH, and the ®ltrate was
concentrated under reduced pressure.
(81 MHz, Me2SO-d6) ꢁ 30.21. Anal. Calcd for
C18H35Cl2N2O6P: C, 45.29; H, 7.39; N, 5.87.
Found: C, 45.37; H, 7.36; N, 5.75.
1-Dodecyl derivative (22).Ð0.15 g (90%); mp
ꢀ
103±105 C; [ꢂ]d 68.0ꢀ (c 1.5, CH2Cl2); FABMS:
1
m/z 555 (100%) (MNa+); H NMR (500 MHz,
2
Me2SO-d6): ꢁ 5.53 (dd, 1-H, J2,NH 2.2, JH,P
16.8 Hz, N-H), 5.16 (d, 1-H, J4,OH 7.1 Hz, OH-4),
4.54 (t, 1-H, J6,OH 5.6 Hz, OH-6), 4.50 (t, 1-H,
J2,3=J3,4 4.3 Hz, H-3), 4.42 (d, 1-H, J1,2 7.2 Hz, H-
1), 3.9-3.1 [m, H-2, H-4, H-5, H-6, H-60,
N(CH2CH2Cl)2, OCH2R], 1.6±1.1 [m, (CH2)10],
0.84 (t, 3H, J 6.8 Hz, CH3); 13C NMR (50 MHz,
Me2SO-d6): ꢁ 103.2 (C-1), 80.0 (C-3), 74.4 (C-5),
68.8 (OCH2R), 65.4, 65.3 (C-4), 60.8 (C-6), 56.0,
55.9 (C-2), 48.6, 48.5 (NCH2), 42.3 (CH2Cl), 31.3,
29.2, 29.0, 28.7, 25.5, 22.1 [(CH2)10], 14.0 (CH3).
31P NMR (81 MHz, Me2SO-d6): ꢁ 30.22. Anal.
Calcd for C22H43Cl2N2O6P: C, 49.53; H, 8.12; N,
5.25. Found: C, 49.58; H, 7.86; N, 5.07.
Cyclꢀohexyl derivative (20).Ð0.12 g (88%); mp
82±84 C; [ꢂ]d 309.1ꢀ (c 0.4, CH2Cl2); FABMS:
Acknowledgement
1
m/z 469 (100%) [MNa+]; H NMR (200 MHz,
2
We thank the DGICYT (Spain) for ®nancial
support (grant PB94-1431).
Me2SO-d6): ꢁ 5.42 (dd, 1-H, J2,NH 2.1, JH,P
16.8 Hz, N-H), 5.16 (d, 1-H, J4,OH 6.3 Hz, OH-4),
4.52 (m, 3-H, H-1, H-3, OH-6), 3.8±3.1 (m), 2.0±
1.1 [m, (CH2)5]; 13C NMR (50 MHz, Me2SO-d6): ꢁ
100.9 (C-1), 80.1 (C-3), 75.5 (OCH), 74.2 (C-5),
65.2, 65.1 (C-4), 60.5 (C-6), 55.9, 55.7 (C-2), 48.4,
48.3 (NCH2), 42.0 (CH2Cl), 32.9, 30.9, 25.0, 23.4,
23.1 [(CH2)5); 31P NMR (81 MHz, Me2SO-d6): ꢁ
30.17. Anal. Calcd for C16H29Cl2N2O6P: C, 42.96;
H, 6.54; N, 6.26. Found: C, 42.84; H, 6.41; N, 6.19.
1-Octyl derivative (21).Ð0.12 g (85%); mp 99±
101 ꢀC; [ꢂ]d 147.6ꢀ (c 1.0, CH3OH); FABMS: m/
z 499 (100%) [MNa+]; 1H NMR (500 MHz,
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2
Me2SO-d6): ꢁ 5.53 (dd, 1-H, J2,NH 2.4, JH,P
16.8 Hz, N±H), 5.16 (d, 1-H, J4,OH 7.1 Hz, OH-4),
4.54 (t, 1-H, J6,OH 5.8 Hz, OH-6), 4.50 (t, 1-H,
J2,3=J3,4 4.3 Hz, H-3), 4.43 (d, 1-H, J1,2 7.2 Hz, H-
1), 3.78 (m, 1-H, OCHHR), 3.65 [m, 5-H, H-6,
(CH2Cl)2], 3.5±3.3 [m, H-4, H-5, H-60, OCHHR,
N(CH2)2], 3.27 (m, 1-H, H-2), 1.5±1.1 [m, (CH2)6],
0.85 (t, 3H, J 6.8 Hz, CH3); 13C NMR (125 MHz,
Me2SO-d6): ꢁ 103.2 (C-1), 79.9 (C-3), 74.3 (C-5),
68.8 (OCH2R), 65.4 (C-4), 60.9 (C-6), 56.0 (C-2),
48.6 (NCH2), 42.3 (CH2Cl), 31.2, 29.1, 28.9, 28.6,
25.5, 22.0 [(CH2)6], 13.9 (CH3); 31P NMR
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unpublished results.
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