Synthesis and biological activity of phosphonate analogs of mannose 6- phosphate (M6P)

Article abstract of DOI:10.1002/(sici)1099-0690(199902)1999:2<447::aid-ejoc447>3.0.co;2-r

Two phosphonate analogs of mannose 6-phosphate (M6P) have been synthesized. The isosteric analog 1 was obtained by a Wittig-Horner reaction at position 6 of a sugar aldehyde. The non-isosteric analog 2 was obtained by a Michaelis-Arbuzov rearrangement of

Full text of DOI:10.1002/(sici)1099-0690(199902)1999:2<447::aid-ejoc447>3.0.co;2-r

FULL PAPER
Synthesis and Biological Activity of Phosphonate Analogs of Mannose
6-Phosphate (M6P)
[a]
[a]
[b]
[a]
`
´
Carole Vidil, Alain Morere,* Marcel Garcia, Veronique Barragan,
Bassou Hamdaoui,[a] Henri Rochefort,[b] and Jean-Louis Montero*[a]
Keywords: Phosphonate analogs / Mannose 6-phosphate (M6P) / M6P receptors / Recognition markers
Two phosphonate analogs of mannose 6-phosphate (M6P)
have been synthesized. The isosteric analog 1 was obtained
by a Wittig–Horner reaction at position 6 of a sugar aldehyde.
The non-isosteric analog 2 was obtained by a Michaelis–
Arbuzov rearrangement of a 6-bromo derivative. In contrast
to the non-isosteric analog 2, the isoster 1 was shown to bind
to M6P receptors as effectively as does M6P itself, thus
demonstrating the considerable potential of the system in
drug design.
Mannose 6-phosphate (M6P) moieties are involved in the phonates is of potential interest. Several methods for the
selective targeting of newly synthesized enzymes to lyso- preparation of sugar phosphonic acids have been described
somes and in the activation of pro-TGFβ. Specific receptors in the literature.[6] In this article we describe the synthesis
recognize M6P residues that are added to the nascent en- of phosphonates 1 and 2 in order to study their affinity
zyme molecule in the Golgi apparatus. ReceptorϪligand toward M6P receptors.
complexes are sorted in a prelysosomal compartment where
they dissociate due to the acidic pH. The ligands are deliv-
ered to lysosomes and the receptor can be recycled to the
Golgi apparatus. Two different M6P receptors have been
Results and Discussion
identified. The larger M6P receptor, also called the M6P/
IGF II receptor since it binds IGF II growth factor, is a
Synthesis of Phosphonate Analogs of M6P
glycoprotein of 275 kDa. The smaller M6P receptor is a
glycoprotein of 46 kDa, which requires divalent cations for
optimal ligand binding. While both receptors cycle to the
plasma membrane, only the M6P/IGF II receptor mediated
the endocytosis of extracellular M6P-containing ligands
(for a review, see Kornfeld[1][2]). Thus, the M6P/IGF II re-
ceptor on the cell surface can be used for targeting exogen-
ous proteins, which possess the M6P signal, to endosomes
and lysosomes. It has already been shown in lysosomal dis-
eases that treatment with an exogenous enzyme could par-
tially restore its deficiency in many organs.[3,4] In addition,
the level of the M6P/IGF II receptor mRNA in vivo is in-
creased in breast cancer tumors as compared to benign
breast disease, a fact that also suggests a possible targeting
of cytotoxic drugs into cancer cells.[5]
The two phosphonate analogs of mannose 6-phosphate
(M6P), shown in Scheme 1, were prepared by two differ-
ent pathways.
For these reasons, the M6P signal seems to be an interest-
ing candidate to target bioactive molecules to endosomes
and thereafter to lysosomes, but the major drawback with
phosphates is their sensitivity to hydrolysis by phosphatases.
In order to alleviate this hydrolysis, it seemed appropriate
to use an M6-phosphonate in which a PϪO bond is re-
placed by a PϪC bond, which is stable toward hydrolases.
Moreover, the possible attachment of drugs to sugar phos-
Scheme 1. Phosphonate analogs of M6P
One pathway led to the isosteric compound 1, while the
other led to a non-isosteric analog 2, which possesses one
less carbon atom between its phosphorus atom and the su-
gar moiety. In both cases, the alcohol 3[7] is the starting
compound (Scheme 1).
[a]
´
´
Laboratoire de Chimie Biomoleculaire CC073, Universite Mont-
pellier II,
`
Place Eugene Bataillon, F-34095 Montpellier cedex 05, France
E-mail: morere@univ-montp2.fr
montero@univ-montp2.fr
For the synthesis of the mannose 6-phosphate isoster 1,
we followed the strategy described in Scheme 2. Intermedi-
[b]
´
Unite Hormones et Cancer (U148), INSERM,
60, rue de Navacelles, F-34090 Montpellier, France
Eur. J. Org. Chem. 1999, 447Ϫ450
WILEY-VCH Verlag GmbH, D-69451 Weinheim, 1999
1434Ϫ193X/99/0202Ϫ0447 $ 17.50ϩ.50/0
447

Synthesis and Biological Activity of Phosphonate Analogs of Mannose 6-Phosphate (M6P)
FULL PAPER
ate 3 was quantitatively oxidized to aldehyde 4[7] by a sphonate 2, a non-isosteric analog of M6P, was obtained
Swern reaction.[8]
after deprotecting phosphonate 9 using trimethylsilyl bro-
mide followed by treatment with a cation exchange resin.
Scheme 3. i: a) CH3SO2Cl, Et3N, CH2Cl2, 30 min, 0°C; b) LiBr,
butanone, 80°C, 1 h 30 min. Ϫ ii: P(OEt)3, 160°C, 6 h.
Ϫ iii: EtOH/H2O (5:1), H2/Pd/C, 20°C, 24 h. Ϫ iv: a)
CH3CN, pyridine, (CH3)3SiBr, 2 h, 20°C; b) H2O, pyri-
dine, 0°C, then room temp., 2 h, cation exchange resin,
5 h, room temp.
Scheme 2. i: (COCl)2, DMSO, iPr2NEt, THF, Ϫ60°C, 20 min. Ϫ
ii: tetraethyl methylenediphosphonate (TEMDP), NaH,
C6H6, 20°C, 30 min. Ϫ iii: EtOH/H2O (5:1), H2/Pd/C,
20°C, 24 h. Ϫ iv: a) CH3CN, pyridine, (CH3)3SiBr, 2 h,
20°C; b) H2O, pyridine 0°C, then room temp., 2 h;
cation exchange resin, 5 h, room temp.
The latter compound, on treatment with the tetraethyl
Biological Activity of Phosphonate Analogs of M6P
methylenediphosphonate carbanion, led (by
a
Wit-
tigϪHorner[9] reaction in 77% yield) to the phosphonate 5,
The affinity of phosphonates 1 and 2 toward the M6P/
which possesses an exocyclic double bond in position 6Ϫ7. IGF II receptor was assayed on pentamannose 6-phosphate
Hydrogenation of phosphonate 5 using a Pd/C catalyst al- Sepharose columns.[13] Retained receptors were eluted using
lowed not only the removal of the benzyl groups at posi- increasing concentrations of the two compounds and ana-
tions 2, 3 and 4, but also the reduction of the double bond. lyzed on silver-stained polyacrylamide gels (Figure 1). The
Derivative 6 was transformed into phosphonate 1 after re- 270-kDa bands eluted from the column correspond to the
action with trimethylsilyl bromide[10] followed by treatment M6P/IGF II receptor. The isosteric phosphonate 1 eluted
with a cation exchange resin.
the receptor in a mannose-dependent concentration with a
The strategy described in Scheme 3 was chosen for the maximum at 10 m. This compound displayed a higher af-
preparation of non-isosteric analog 2. Derivative 3 was finity than the non-isosteric derivative 2 since this last com-
quantitatively mesylated in position 6 and then transformed pound only partially eluted the receptor. Interestingly, the
into the bromide by heating in the presence of LiBr in but- receptor remaining on the affinity column after addition
anone, thereby generating adduct 7[11] in 90% yield. The of compound 2 could be further dissociated with 10 m
conversion into the phosphonate 8 was effected by a phosphonate 1 (Figure 1, column 7).
MichaelisϪArbusov reaction in which the bromide deriva-
Phosphonate 1 was found to have a similar affinity to-
tive was heated at 160°C with triethyl phosphite.[12] The ward the M6P/IGF II receptors as the mannose 6-phos-
expected compound 8 was obtained in 75% yield. The phos- phate routinely used at 10 m to elute this receptor (data
phonate was then deprotected in two stages: The secondary not shown). On the other hand, the non-isosteric analog of
alcohol functions at positions 2, 3 and 4 were first released M6P, 2, shows only a weak affinity toward those receptors.
by hydrogenolysis (H2, Pd/C) to give compound 9. The pho- Thus, the replacement of the oxygen atom by a methylene
448
Eur. J. Org. Chem. 1999, 447Ϫ450

`
A. Morere, J.-L. Montero et al.
FULL PAPER
amine, then reduced with sodium tetrahydroborate, and finally cou-
pled on active Sepharose to lead to pentamannan 6-phosphate Se-
pharose.[13] Ϫ SDS polyacrylamide gel electrophoresis was per-
formed according to Laemmli[15] and proteins visualized using a
silver-staining kit (Biorad).
Methyl 2,3,4-Tri-O-benzyl-6-deoxy-6-diethoxyphosphinylmethylene-
␣--mannopyranoside (5): 95% NaH (0.087 g, 3.46 mmol) was ad-
ded to 24 mL of anhydrous benzene under nitrogen. 1.07 mL (4.32
mmol) of tetraethyl methylenediphosphonate was added dropwise
and 30 min later 0.8 g (1.73 mmol) of 4 in 7.2 mL of anhydrous
benzene was added dropwise at 20°C. After stirring at room temp.
for 30 min, the benzene was removed. The residue was dissolved in
CH2Cl2 and water was added. The organic layer was extracted,
dried and concentrated. Compound 5 was obtained after chroma-
tography on silica gel (hexane/AcOEt, 6:4, then AcOEt) in 77%
yield. Ϫ [α]D ϭ ϩ67.3 (c ϭ 1.07, CHCl3). Ϫ 1H NMR (CDCl3):
δ ϭ 1.36 (t, 6 H, J ϭ 7.1 Hz, 2 ϫ CH3CH2OP), 3.33 (s, 3 H,
OCH3), 3.71 (t, 1 H, J3,4 ϭ J4,5 ϭ 9.3 Hz, 4-H), 3.83 (dd, 1 H,
J1,2 ϭ 1.7 Hz, J2,3 ϭ 3.1 Hz, 2-H), 3.94 (dd, 1 H, 3-H), 4.04Ϫ4.25
(m, 5 H, 2 ϫ CH3CH2OP and 5-H), 4.63 and 4.93 (AMq, 2 H,
Figure 1. Analysis on electrophoresis gel of retained receptors
eluted with compounds 1 and 2; positions of standard molecular
weights are indicated
J
AM ϭ 10.6 Hz, CH2Ph), 4.67 (s, 2 H, CH2Ph), 4.74 and 4.82 (ABq,
group does not modify the recognition of the substrate by
the receptor.
JAB ϭ 12.4 Hz, CH2Ph), 4.77 (d, 1 H, 1-H), 6.17 (ddd, 1 H, J5,7
ϭ
1.7 Hz, J6,7 ϭ 17.3 Hz, J7,P ϭ 21.3 Hz, 7-H), 7.01 (ddd, 1 H, J5,6 ϭ
4.3 Hz, J6,P ϭ 21.6 Hz, 6-H), 7.32Ϫ7.42 (m, 15 H, 3 C6H5). Ϫ 13C
NMR (CDCl3): δ ϭ 16.4 (m, 2 ϫ CH3CH2OP), 55.0 (OCH3), 61.8
(m, 2 ϫ CH3CH2OP), 71.3 (d, JC,P ϭ 21.3 Hz, C-5), 72.4, 73.0,
75.4 (3 ϫ CH2Ph), 74.8, 78.3, 80.2 (C-2,3,4), 99.4 (C-1), 118.1 (d,
JC,P ϭ 188.3 Hz, C-7), 127.7Ϫ128.4 (CH arom.), 138.1, 138.3,
138.4 (3 ϫ Cq), 148.1 (d, JC,P ϭ 5.8 Hz, C-6). Ϫ 31P NMR (CDCl3):
δ ϭ 14.51. Ϫ MS (FABϩ); m/z (%): 619 (1) [M ϩ Na]ϩ, 597 (7) [M
ϩ H]ϩ. Ϫ C33H41O8P (596.65): calcd. C 66.43, H 6.93; found C
Conclusion
In summary, we have prepared two phosphonate analogs
of M6P. The isosteric analog binds to the M6P/IGF II re-
ceptor with an affinity higher than the non-isosteric deriva-
tive and similar to M6P. This suggests the considerable po-
tential of such compounds in targeting therapy. Thus, the 66.41, H 6.95.
addition of isosteric mannose 6-phosphonate on recombi-
Methyl 6-Deoxy-6-diethoxyphosphinylmethyl-␣--mannopyranoside
nant enzymes could be used in substitutive therapy of lyso-
somal disease. The synthesis of isosteric M6P analogs con-
jugated to drugs and other moieties of biological interest is
now being considered in relation to targeted therapy in tis-
sues rich in M6P/IGF II receptors.
(6): To a solution of 5 (1.38 g, 2.31 mmol) in 350 mL of EtOH/
H2O (5:1) was added 0.138 g of 10% Pd/C. After stirring under
hydrogen for 24 h, the mixture was filtered through Celite and then
concentrated under vacuum. The residue was purified by chroma-
tography on silica gel (AcOEt/MeOH, 9:1) to give 6 in 93% yield.
Ϫ [α]D ϭ ϩ54.4 (c ϭ 1.47, CHCl3). Ϫ 1H NMR (CDCl3): δ ϭ 1.35
(t, 6 H, J ϭ 7.2 Hz, 2 ϫ CH3CH2OP), 1.71Ϫ2.25 (m, 4 H, 6,6Ј,7,7Ј-
H), 2.41 (s, 1 H, OH), 3.35 (s, 3 H, OCH3), 3.41Ϫ3.50 (m, 1 H, 2-
Experimental Section
H), 3.51 (t, 1 H, J3,4 ϭ J4,5 ϭ 8.9 Hz, 4-H), 3.75 (dd, 1 H, J2,3
3.2 Hz, 3-H), 3.88Ϫ3.98 (m, 1 H, 5-H), 4.03Ϫ4.21 (m, 4 H, 2 ϫ
CH3CH2OP), 4.45Ϫ4.65 (m, 2 H, 2 ϫ OH), 4.72 (d, 1 H, J1,2
ϭ
General Aspects: Reactions were monitored by TLC using alu-
minium-coated plates with silica gel 60 F254 (Merck) and visualized
by UV light and/or by charring with H2SO4 (aqueous 10% spray
solution). Aldehydes were developed by spraying with a 5% rhod-
anine solution in ethanol. Molybdenum blue was used to develop
phosphorus-containing compounds. Ϫ Column chromatography
ϭ
1.4 Hz, 1-H). Ϫ 13C NMR (CDCl3): δ ϭ 16.4 (m, 2 ϫ CH3CH2O),
21.0 (d, JC,P ϭ 142.2 Hz, C-7), 24.2 (C-6), 54.9 (OCH3), 62.0 (m,
2 ϫ CH3CH2OP), 70.6, 70.7, 71.8 (C-2,3,4), 71.3 (d, JC,P ϭ 15.7
Hz, C-5), 101.0 (C-1). Ϫ 31P NMR (CDCl3): δ ϭ 34.05. Ϫ MS
(FABϩ); m/z (%): 351 (45) [M ϩ Na]ϩ, 329 (85) [M ϩ H]ϩ. Ϫ
C12H25O8P (328.29): calcd. C 43.90, H 7.68; found C 43.91, H 7.66.
1
was performed with Merck silica gel 60 H (Art. 9385). Ϫ H- and
31P-NMR spectra were recorded with an AC-250 Bruker spec-
trometer and 13C-NMR spectra with a WP-200-SY Bruker spec-
trometer. Chemical shifts are given on the δ scale using the residual
solvent peaks as a reference relative to TMS. Ϫ Specific rotations
were measured with sodium-D light at 20°C using a PerkinϪElmer
spectrometer. Ϫ Microanalyses were performed in the microanal-
Methyl 6-Deoxy-6-dihydroxyphosphinylmethyl-␣--mannopyrano-
side Sodium Salts 1: To a solution of 6 (0.65 g, 2.0 mmol) in 25
mL of anhydrous CH3CN under nitrogen was added 0.25 mL (0.42
mol) of pyridine and 2.6 mL (20 mmol) of Me3SiBr. After stirring
ysis laboratory of ENSCM (Montpellier). Ϫ Mass spectra were at room temp. for 2 h, 15 mL of distilled water and 0.42 mL of
measured with a DX 300 JEOL spectrometer in the FABϩ ion
pyridine (5.25 mmol) were added at 0°C. Stirring was continued at
mode. Ϫ Compounds 3, 4 and 7 were prepared according to pub-
room temp. for 2 h, then the aqueous phase was washed with
lished procedures.[7,11] Ϫ The cation exchange resin (50WX2 Hϩ CH2Cl2. The organic layer was then concentrated before the ad-
Dowex) was washed with a 1  NaOH solution and then with
distilled water before use. Ϫ The M6P/IGF II receptor was purified
from fetal calf serum as previously described.[14] The pentaman-
nose 6-phosphate was functionalized with β-(p-aminophenol)ethyl-
dition of 60 mL of distilled water and 30 g of cation exchange resin.
After stirring at room temp. for 5 h, the resin was filtered off and
washed several times with water. The aqueous phase was concen-
trated and the residue was purified by chromatography on reversed
Eur. J. Org. Chem. 1999, 447Ϫ450
449

Synthesis and Biological Activity of Phosphonate Analogs of Mannose 6-Phosphate (M6P)
FULL PAPER
1
phase (RP-18, H2O). After lyophylization, 1 was obtained in 65% MeOH). Ϫ H NMR (D2O): δ ϭ 1.75 (ddd, 1 H, J5,6Ј ϭ 9.5 Hz,
yield. Ϫ [α]D ϭ ϩ61.9 (c ϭ 1.05, CH3OH). Ϫ 1H NMR (D2O): J6,6Ј ϭ 15.5 Hz, J6Ј,P ϭ 15.5 Hz, 6Ј-H), 2.10 (ddd, 1 H, J5,6 ϭ 2.9
δ ϭ 1.50Ϫ2.10 (m, 4 H, 6,6Ј,7,7Ј-H), 3.28 (s, 3 H, OCH3),
3.30Ϫ3.50 (m, 2 H, 4,5-H), 3.65 (dd, 1 H, J3,2 ϭ 3.4 Hz, J3,4 ϭ 9.4
Hz, 3-H), 3.79 (dd, 1 H, J1,2 ϭ 1.6 Hz, 2-H), 4.60 (d, 1 H, 1-H).
Hz, J6,P ϭ 18.5 Hz, 6-H), 3.35 (s, 3 H, OCH3), 3.38 (t, 1 H, J3,4 ϭ
J4,5 ϭ 9.6 Hz, 4-H), 3.63 (dd, 1 H, J2,3 ϭ 3.4 Hz, 3-H), 3.72 (dd,
1 H, 5-H), 3.80 (dd, 1 H, J1,2 ϭ 1.7 Hz, 2-H), 4.60 (d, 1 H, 1-H).
Ϫ
13C NMR (D2O): δ ϭ 31.3 (d, JC,P ϭ 134.7 Hz, C-6), 55.8
Ϫ
13C NMR (D2O): δ ϭ 23.6 (d, JC,P ϭ 136.6 Hz, C-7), 25.2 (d,
JC,P ϭ 3.8 Hz, C-6), 55.4 (OCH3), 70.6, 70.8, 71.2 (C-2,3,4), 72.6 (OCH3), 69.4 (d, JC,P ϭ 4.7 Hz, C-5), 70.7(C-2), 71.1 (d, JC,P
ϭ
(d, JC,P ϭ 17.1 Hz, C-5), 101.6 (C-1). Ϫ 31P NMR (D2O): δ ϭ
31.04. Ϫ MS (FABϪ ); m/z (%): 271 (55) [M Ϫ 2 Na ϩ H]Ϫ. Ϫ
C8H15Na2O8P (316.15): calcd. C 30.39, H 4.78; found C 30.41, H
4.83.
2.1 Hz, C-3), 72.2 (d, JC,P ϭ 12.9 Hz, C-4), 101.4 (C-1). Ϫ 31P
NMR (D2O): δ ϭ 24.46. Ϫ MS (FABϪ ); m/z (%): 257 (60) [M Ϫ
2 Na ϩ H]Ϫ. Ϫ C7H13Na2O8P (302.12): calcd. C 27.83, H 4.34;
found C 27.78, H 4.37.
Methyl 2,3,4-Tri-O-benzyl-6-deoxy-6-diethoxyphosphinyl-␣--man-
nopyranoside (8): Compound 7 (2.50 g, 4.75 mmol) was dissolved
in 35 mL of P(OEt)3 under nitrogen. After stirring at 160°C for 6
h, P(OEt)3 was removed by distillation and the product was puri-
fied by chromatography on silica gel (hexane/AcOEt, 6:4, then Ac-
OEt) to give compound 8 in 75% yield. Ϫ [α]D ϭ ϩ21.6 (c ϭ 1.11,
CHCl3). Ϫ 1H NMR (CDCl3): δ ϭ 1.36 (td, 6 H, J ϭ 7.1 Hz,
JCH3ϪP ϭ 1 Hz, 2 CH3CH2OP), 2.01 (ddd, 1 H, J5,6Ј ϭ 10.4 Hz,
Acknowledgments
We are extremely grateful to C. Rougeot for the biological pro-
cedures and to Dr. F. M. Menger for helpful discussions and for
critical reading of the manuscript.
[1]
S. Kornfeld, I. Mellman, Annu. Rev. Cell Biol. 1989, 5,
J6,6Ј ϭ J6Ј,P ϭ 15.4 Hz, 6Ј-H), 2.38 (ddd, 1 H, J5,6 ϭ 1.8 Hz, J6,6Ј
ϭ
483Ϫ525.
[2]
S. Kornfeld, Annu. Rev. Biochem. 1992, 61, 307Ϫ330.
15.4 Hz, J6,P ϭ 20.1 Hz, 6-H), 3.41 (s, 3 H, OCH3), 3.68 (t, 1 H,
J3,4 ϭ J4,5 ϭ 9.4 Hz, 4-H), 3.81 (dd, 1 H, J1,2 ϭ 1.7 Hz, J2,3 ϭ 3.0
Hz, 2-H), 3.90 (dd, 1 H, 3-H), 3.98Ϫ4.00 (m, 1 H, 5-H), 4.01Ϫ4.21
(m, 4 H, 2 ϫ CH3CH2OP), 4.61 (s, 2 H, CH2Ph), 4.65 and 5.05
(AMq, 2 H, JAM ϭ 11.2 Hz, CH2Ph), 4.68 (d, 1 H, 1-H), 4.71 and
4.79 (ABq, 2 H, JAB ϭ 12.5 Hz, CH2Ph), 7.21Ϫ7.52 (m, 15 H, 3
C6H5). Ϫ 13C NMR (CDCl3): δ ϭ 16.3 (m, 2 ϫ CH3CH2OP), 28.4
(d, JC,P ϭ 142.3 Hz, C-6), 55.1 (OCH3), 61.6 (m, 2 ϫ CH3CH2OP),
67.2 (d, JC,P ϭ 6.7 Hz, C-5), 74.9 (C-2), 78.7 (d, JC,P ϭ 45.6 Hz,
C-4), 80.3 (d, JC,P ϭ 3.3 Hz, C-3), 72.2, 73.0, 74.9 (3 ϫ CH2Ph),
99.1 (C-1), 127.6Ϫ128.4 (CH arom.). Ϫ 31P NMR (CDCl 3): δ ϭ
30.11. Ϫ MS (FABϩ); m/z (%): 585 (6) [Mϩ ϩ H]. Ϫ C32H41O8P
(584.64): calcd. C 65.74, H 7.07; found C 65.80, H 7.06.
[3]
E. Beutler, A. Kay, A. Saven, P. Garver, D. Thurstonn, A. Daw-
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Methyl 6-Deoxy-6-diethoxyphosphinyl-␣--mannopyranoside (9):
The procedure described for the preparation of 6 was employed to
1
[9]
obtain 9 in 90% yield. Ϫ [α]D ϭ ϩ55.3 (c ϭ 1.03, CHCl3). Ϫ H
G. H. Jones, E. K. Hamamura, J. G. Moffatt, Tetrahedron Lett.
1968, 55, 5731Ϫ5734; K. Yoshino, T. Kono, T. Morita, G. Tsu-
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NMR (CDCl3): δ ϭ 1.32 (t, 6 H, J ϭ 7.1 Hz, 2 ϫ CH3CH2OP),
1.88 (s, 2 H, 2 ϫ OH), 2.13Ϫ2.51 (m, 2 H, CH2P), 3.42 (s, 3 H,
OCH3), 3.70 (t, 1 H, J3,4 ϭ J4,5 ϭ 9.2 Hz, 4-H), 3.75Ϫ4.00 (m, 4
H, 2,3,4-H, OH), 4.04Ϫ4.20 (m, 4 H, 2 ϫ CH3CH2OP), 4.69 (s, 1
H, J1,2 ϭ 1.2 Hz, 1-H). Ϫ 13C NMR (CDCl3): δ ϭ 16.4 (m, 2 ϫ
CH3CH2O), 28.7 (d, JC,P ϭ 141.6 Hz, C-6), 55.2 (OCH3), 62.1 (m,
2 ϫ CH3CH2OP), 67.5 (d, JC,P ϭ 4.2 Hz, C-5), 70.6, 71.6 (C-2,3),
71.7 (d, JC,P ϭ 8.4 Hz, C-4), 101.0 (C-1). Ϫ 31P NMR (CDCl3):
δ ϭ 31.40. Ϫ MS (FABϩ); m/z (%): 337 (97) [M ϩ Na]ϩ, 315 (100)
[M ϩ H]ϩ. Ϫ C11H23O8P (314.27): calcd. C 42.04, H 7.38; found
C 42.02, H 7.39.
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Methyl 6-Deoxy-6-dihydroxyphosphinyl-␣--mannopyranoside So-
dium Salts 2: The procedure described for the preparation of 1 was
employed to obtain 2 in 65% yield. Ϫ [α]D ϭ ϩ19.2 (c ϭ 1.51,
Received July 28, 1998
[O98350]
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