Unified total syntheses of the inositol polyphosphates: D-I-3,5,6P 3, D-I-3,4,5P3, D-I-3,4,6P3, and D-I-3,4,5,6P4 via catalytic enantioselective and site-selective phosphorylation

Article abstract of DOI:10.1021/jo0610816

Synthetic routes to various inositol polyphosphates have been discovered utilizing catalytic enantioselective and site-selective phosphorylation reactions. The syntheses described herein exploit a common intermediate to gain efficient access to eight unique inositol polyphosphates.

Full text of DOI:10.1021/jo0610816

Unified Total Syntheses of the Inositol Polyphosphates: D-I-3,5,6P3,
D-I-3,4,5P3, D-I-3,4,6P3, and D-I-3,4,5,6P4 via Catalytic
Enantioselective and Site-Selective Phosphorylation
Adam J. Morgan, Shio Komiya, Yingju Xu, and Scott J. Miller*,†
Department of Chemistry, Boston College, Chestnut Hill, Massachusetts 02467
scott.miller@yale.edu
ReceiVed May 26, 2006
Synthetic routes to various inositol polyphosphates have been discovered utilizing catalytic enantioselective
and site-selective phosphorylation reactions. The syntheses described herein exploit a common intermediate
to gain efficient access to eight unique inositol polyphosphates.
Introduction
effectively desymmetrized compound 1 to deliver either mono-
phosphate 2 or ent-2 in essentially optically pure form. In more
recent studies, we applied these catalysts to the synthesis of
members of the phosphatidylinositol class of natural products.6
This work culminated in the synthesis of phosphatidylinositol-
3-phosphate and phosphatidylinositol-3,5-bis(phosphate), as well
as related analogues. In several respects, application of these
The inositol phosphates comprise a class of biologically active
molecules implicated in signal transduction pathways throughout
biochemistry.1 D-myo-Inositol-1,4,5-trisphosphate (D-I-1,4,5P3),
for example, has been shown to serve as a secondary messenger
modulating the concentration of Ca2+ in cells.2 For this reason,
D-I-1,4,5-P3 (L-I-3,5,6P3) has become one of the most thoroughly
studied molecules in this class. Although these and other inositol
phosphates have been the subject of many synthetic and
biochemical research endeavors, in-depth studies could be
greatly expanded if synthetic access to these compounds and
their analogues could be improved.3 In that regard, concise
synthetic routes to several of the inositol polyphosphates,
including D-I-3,5,6P3 (L-I-1,4,5-P3), employing peptide-catalyzed
asymmetric phosphorylation are presented herein (Figure 1).
We anticipated that our recently described enantiodivergent
syntheses of D-I-1P and D-I-3P could be used as a conduit to
the synthesis of many of the inositol polyphosphates (Figure
2).4,5 In this work, we discovered peptides 1P and 3P that
(3) (a) Baker, R.; Kulagowski, J. J.; Billington, D. C.; Leeson, P. D.;
Lennon, I. C.; Liverton, N. J. J. Chem. Soc., Chem. Commun. 1989, 1383-
1385. (b) Baker, R.; Carrick, C.; Leeson, P. D.; Lennon, I. C.; Liverton, N.
J. J. Chem. Soc., Chem. Commun. 1991, 298-300. (c) Bone, R.; Frank, L.;
Springer, J. P.; Pollack, S. J.; Osbourne, S. A.; Atack, J. R.; Knowles, M.
R.; McAllister, G.; Ragan, C. I.; Broughton, H. B.; Baker, R.; Fletcher, S.
R. Biochemistry 1994, 33, 9460-9467. (d) Miller, D. J.; Beaton, M. W.;
Wilkie, J.; Gani, D. ChemBioChem 2000, 1, 262-271. (e) Baker, R.; Leeson,
P. D.; Liverton, N. J.; Kulagowski, J. J. J. Chem. Soc., Chem. Commun.
1990, 462-464. (f) Kozikowski, A. P.; Qiao, L.; Tuckmantel, W.; Powis,
G. Tetrahedron 1997, 53, 14903-14914.
(4) (a) Sculimbrene, B. R.; Miller, S. J. J. Am. Chem. Soc. 2001, 123,
10125-10126. (b) Sculimbrene, B. R.; Morgan, A. J.; Miller, S. J. Am.
Chem. Soc. 2002, 124, 11653-11656. (c) Sculimbrene, B. R.; Morgan, A.
J.; Miller, S. J. Chem. Commun. 2003, 1781-1785.
(5) For recent, general, and divergent approaches to various inositol
polyphosphates, see: (a) Podeschwa, M. A. L.; Plattenburg, O.; Altenbach,
H.-J. Eur. J. Org. Chem. 2005, 3101-3127. (b) Takahashi, H.; Kittaki, H.;
Ikegami, S. J. Org. Chem. 2001, 66, 2705-2716. (c) Leung, L. W.; Bittman,
R. Carbohydr. Res. 1998, 305, 171-179.
(6) (a) Sculimbrene, B. R.; Xu, Y.; Miller, S. J. J. Am. Chem. Soc. 2004,
126, 13182-13183. (b) Xu, Y.; Sculimbrene, B. R.; Miller, S. J. J. Org.
Chem. 2006, 71, 4919-4928.
† Present address: Department of Chemistry, Yale University, P.O. Box
208107, New Haven, CT 06520.
(1) (a) Irvine, R. F.; Schell, M. J. Nature ReV. Mol. Cell. Biol. 2001, 2,
327-338. (b) Billington, D. C. The Inositol Phosphates: Chemical Synthesis
and Biological Significance; VCH: New York, 1993.
(2) (a) Hokin, M. R.; Hokin, L. E. J. Biol. Chem. 1953, 203, 967-977.
(b) Mitchell, R. H. Biochim. Biophys. Acta 1975, 415, 81-147.
10.1021/jo0610816 CCC: $33.50 © 2006 American Chemical Society
Published on Web 07/29/2006
J. Org. Chem. 2006, 71, 6923-6931
6923

Morgan et al.
TABLE 1. Survey of Protecting Groups for Phosphorylation
SCHEME 1
SCHEME 2a
ee (%)
entry
R1
Bn
Bn
PMB
Bn
R2
Bn
allyl
PMB
PMB
peptide 1P
peptide 3P
1
2
3
4
98
94
98
97
-98
-97
-98
-98
a (a) peptide 3P, DPCP, Et3N, CH2Cl2, 53%, 98% ee; (b) (i) BOMCl,
Hunig’s base, DMF; (ii) NaH, BnOH, 51% for two steps; (c) DDQ, CH2Cl2,
80%; (d) (i-Pr)2NP(OBn)2, dicyanoimidazole, H2O2, 78%; (e) Pd(OH)2/C,
H2, 88%.
(PMB) substituents were suitable for obtaining the products in
high ee (94-98% ee). 2-O-Benzyl-4,6-bis(p-methoxybenzyl)-
myo-inositol (entry 4) emerged as a particularly useful com-
pound, as high enantioselectivity was observed within a substrate
that possesses orthogonal protection between the 2- and the 4,6-
positions. Therefore, this triol was ultimately chosen as the
common intermediate that would deliver various inositol poly-
phosphates (entry 4).
Our work thus began with the identification of a common
precursor upon which we might devise schemes for the synthesis
of multiple inositol polyphosphates. Our strategy began with a
multigram synthesis of 2-O-benzyl-4,6-bis(p-methoxybenzyl)-
D-myo-inositol (3), which was achieved in a manner similar to
that for tribenzyl inositol (1) (Scheme 1).7
FIGURE 1. Inositol polyphosphates targeted for synthesis.
catalysts to the synthesis of various inositol polyphosphates is
a more challenging objective. For example, to achieve the
synthesis of unique polyphosphates with positional selectivity
on the myo-inositol ring, we required a suitably protected
derivative of 1 such that positions 4, 5, and 6 could be accessed
for functionalization at a later stage.
Results and Discussion
To identify a proper analogue, a series of differentially
protected myo-inositol derivatives were screened as substrates
for the asymmetric phosphorylation using peptide 1P and peptide
3P (Table 1). This screen revealed that catalysts 1P and 3P
maintain high enantioselectivity profiles with a variety of
protecting groups at the 4- and 6-positions. As shown in entries
1-4, various arrays of benzyl, allyl, and p-methoxybenzyl
With an appropriately protected derivative of myo-inositol
identified (3), we began investigating synthetic strategies toward
D-I-3,4,6P3 (L-I-1,4,6P3), which would pave the way for other
polyphosphates (Scheme 2). Asymmetric phosphorylation of
triol 3 was then achieved using peptide 3P, affording mono-
FIGURE 2. Peptide-catalyzed phosphorylation.
6924 J. Org. Chem., Vol. 71, No. 18, 2006

Unified Syntheses of IPn’s
SCHEME 3a
a (a) TBSCl, imidazole, DMF; (b) NaH, BnOH; 79%, two steps; (c) cat. AcCl, MeOH; (d) (i-Pr)2NP(OBn)2, dicyanoimidazole, H2O2; 43%, two steps; (e)
DDQ, CH2Cl2/H2O 67%; (f) (i-Pr)2NP(OBn)2, dicyanoimidazole, H2O2; 4:1 regioselectivity, 70% isolated; (g) Pd(OH)2/C, H2, 97%.
phosphate 4 in 53% yield with 98% ee. Protection of the 1-
and 5-hydroxyl groups as BOM ethers followed by transesteri-
fication of the diphenyl phosphate provides dibenzyl phosphate
5. It is important to note that all attempts to facilitate PMB
deprotection in the presence of an adjacent diphenyl phosphate
results in phosphate migration.8 Deprotection of the PMB ethers
was then accomplished with dichlorodicyanobenzoquinone
(DDQ) affording 4,6-diol 6. The synthesis of D-I-3,4,6P3 (L-I-
1,4,6P3) was then completed by simultaneous phosphorylation
of the 4- and 6-hydroxyl groups followed by global hydro-
genolysis with Pd(OH)2 to give the target, isolated as the sodium
salt (Scheme 2).
With the development of a successful synthesis of one of
the inositol triphosphates, we turned our focus to the synthesis
of D-I-3,4,5P3 (L-I-1,5,6P3). The approach is reminiscent of the
synthetic plan described in Scheme 2, but this case requires a
critical differentiation of the 4- and 6-hydroxyl groups. Our
approach utilizes a TBS group at the 1-position of diol 10
(Scheme 3), which created the expectation that the steric and
electronic environments would be sufficiently different such that
preferential reaction would occur at either the 4- or the
6-position. Thus, compound 4 was subjected to per-silylation,
followed by phosphate ester transesterification to deliver
intermediate 8 (79% yield; 2 steps). Selective hydrolysis of the
5-TBS ether, followed by conventional phosphorylation gave
9, which was then exposed to DDQ to give key diol 10. Notably,
the desilylation of the TBS-protected ether at the 5-position of
compound 8 is faster than the TBS-ether at the 1-position. Thus,
compound 19 is produced in 49% yield, along with recovered
starting material (14%) and diol (23%). Perhaps surprisingly,
isolation of the regioisomeric TBS-protected ether A, resulting
from desilylation at the 1-position is difficult, because of its
formation in minute quantities. Since the 5-position is presumed
to be sterically more encumbered, we presume that there may
be issues of steric relief that lead to selectivity for this step. At
the same time, the free hydroxyl group of 19 may provide
cooperativity in the desilylation of the TBS-ether at the
1-position, leading to the observation.
In the key phosphorylation event, exposure of 10 to phos-
phoramidite coupling conditions led to the formation of 11, with
4:1 regioselectivity.9 Our current contention is that this is a
sterically controlled process, with the spatial requirements of
the TBS ether exceeding that of the dibenzyl phosphate group.
Isolation of the major product was then followed by hydro-
genolysis to deliver D-I-3,4,5P3 (L-I-1,5,6P3). Notably, the TBS-
ether at the 1-position appears to undergo autodeprotection once
the phosphoric diacid functionality is revealed.
Our synthetic strategy outlined for D-I-3,4,5P3 (L-I-1,5,6P3)
stimulated a parallel approach to D-I-3,4,5,6P4 (L-I-1,4,5,6P4,
Scheme 4). This synthesis was initiated with selective desily-
lation of common intermediate 8 followed by PMB-group
deprotection with DDQ to afford 4,5,6-triol 12. Exhaustive
phosphorylation and subsequent global hydrogenolysis rapidly
provided D-I-3,4,5,6P4 (L-I-1,4,5,6P4).
We then turned our attention to the synthesis of D-I-3,5,6P3
(L-I-1,4,5P3). Analysis of our synthesis of D-I-3,4,5P3 (L-I-1,5,-
6P3) led to the hypothesis that the sterically demanding TBS
ethers were responsible for direction of the observed regiose-
lectivity in the phosphorylation of 10 to deliver 11. We therefore
examined 1-OH substituents that are less sterically demanding.
Thus, substrate 13 was synthesized employing a benzyloxy
methyl ether (BOM) protecting group in place of TBS as
outlined in Scheme 5. In this case, the reaction produced a
(7) (a) Lee, H.; Kishi, Y. J. Org. Chem. 1985, 50, 4402-4404. (b)
Baudin, G.; Gluenzer, B. I.; Swaminathan, K. S.; Vasella, A. HelV. Chim.
Acta 1988, 71, 1367-1378. (c) Riley, A. M.; Mahon, M. F.; Potter, B. V.
L. Angew. Chem., Int. Ed. 1997, 36, 1472-1474. (d) Chung, S.-K.; Chang,
Y.-T.; Bioorg. Med. Chem. Lett. 1997, 7, 2715-2718. (e) Sureshan, K.
M.; Watanabe, Y. Tetrahedron: Asymmetry 2004, 15, 1193-1198.
(8) The migratory behavior of diphenyl phosphates within the inositol
framework is known; see ref 1b.
(9) Notably, catalytic P(V)-phosphorylation conditions with substrate 10
failed to give appreciable yields of either regioisomeric product.
J. Org. Chem, Vol. 71, No. 18, 2006 6925

Morgan et al.
SCHEME 4a
a (a) TBSCl, imidazole, DMF; (b) NaH, BnOH; 79%, two steps (c) cat. AcCl, MeOH; (d)DDQ, CH2Cl2/H2O 43%, two steps; (e) (i-Pr)2NP(OBn)2,
Dicyanoimidazole, H2O2; (f) Pd(OH)2/C, H2, 71%, two steps.
SCHEME 5a
a (a) BOMCl, Hunig’s base, THF; (b) (i-Pr)2NP(OBn)2, dicyanoimidazole, H2O2; 29%, two steps; (c) NaH, BnOH; 38%; (d) DDQ, CH2Cl2/H2O, 83%;
(e) (i-Pr)2NP(OBn)2, dicyanoimidazole, H2O2; 53% isolated; (f) Pd(OH)2/C, H2, 90%.
mixture of the monoprotected BOM-ethers, along with bis-
(protected) material as well as recovered starting material.
Although compound 13 is isolated in only 29% yield after two
steps, the desired monoprotected material is the major product
of the BOM-protection reaction (∼2:1 over the alternative
regioisomer), and this procedure proved operationally conve-
nient. Nonetheless, desymmetrized diol 4, upon treatment with
BOMCl and subsequent phosphorylation, affords diphosphate
13. Transesterification of the diphenyl phosphate followed by
PMB deprotection with DDQ provides 4,6-diol 15. Phospho-
rylation of 15 under standard conditions provides 3,5,6-
trisphosphate 16 as a single regioisomer, supporting our
hypothesis for selectivity. Global hydrogenolysis with Pd(OH)2
ultimately provides D-I-3,5,6,P3.
lective manipulation of the desymmetrized ring was achieved
through selective functionalization of the remaining hydroxyl
groups. In all, eight unique myo-inositol polyphosphates are
accessible via this approach. Further studies directed toward
the development of inositol polyphosphate analogues as well
as studies of their behavior on biological systems are ongoing.10
Experimental Section
2-O-Benzyl-4,6-di-O-p-methoxybenzyl-D-myo-inositol (3). To
a solution of 4,6-di-O-p-methoxybenzyl-myo-inositol-1,3,5-O-ortho-
formate11 (930 mg, 2.16 mmol) in DMF (30 mL) was added sodium
hydride (104 mg, 4.32 mmol). The reaction was stirred at room
temperature for 15 min at which time benzyl bromide (257 µL,
2.16 mmol) was added. The reaction continued to stir at room
temperature for 2 h and was then quenched by slow addition of
water. The reaction was then concentrated in vacuo and extracted
with dichloromethane (3 × 200 mL) and washed repeatedly with
water followed by brine. The organic layers were then dried
(MgSO4), filtered, and concentrated to afford 2-O-benzyl-4,6-di-
O-p-methoxybenzyl-D-myo-inositol-1,3,5-O-orthoformate as a clear
Conclusions
The syntheses of four inositol polyphosphates (D-I-3,4,6P3,
D-I-3,4,5P3, D-I-3,4,5,6P4, and D-I-3,5,6P3) has thus been
achieved with excellent efficiency from common intermediate
4. This intermediate is obtained through desymmetrization of 3
by peptide-catalyzed asymmetric phosphorylation employing
peptide 3P. Alternatively, peptide 1P provides access to ent-4
with high levels of selectivity, allowing formal access to each
enantiomer of the inositol polyphosphates described above (D-
I-1,4,6P3, D-I-1,5,6P3, D-I-1,4,5,6P4, and D-I-1,4,5P3). Regiose-
(10) (a) Morgan, A. J.; Wang, Y. K.; Roberts, M. F.; Miller, S. J. J. Am.
Chem. Soc. 2004, 126, 15370-15371. (b) Wang, Y. K.; Morgan, A. J.;
Stieglitz, K.; Stec, B.; Thompson, B.; Miller, S. J.; Roberts, M. F.
Biochemistry 2006, 45, 3307-3314.
(11) Riley, A. M.; Guedat, P.; Schlewer, G.; Spiess, B.; Potter, B. V. L.
J. Org. Chem. 1998, 63, 295-305.
6926 J. Org. Chem., Vol. 71, No. 18, 2006

Unified Syntheses of IPn’s
1
oil (1.03 g, 92%). H NMR (CDCl3, 400 MHz) δ 7.40-7.26 (m,
8.5 Hz, 2H), 6.71 (d, J ) 8.5 Hz, 2H), 4.99 (s, 2H), 4.87-4.48 (m,
13H), 4.29 (br. s, 1H), 4.07 (t, J ) 9.4 Hz, 1H), 3.99 (t, J ) 9.6
Hz, 1H), 3.78-3.65 (m, 1H), 3.74 (s, 3H overlaps with multiplet
at 3.78), 3.71 (s, 3H overlaps with multiplet at 3.78), 3.61 (t, J )
9.2 Hz, 2H); 13C NMR (CDCl3, 100 MHz) δ 158.8, 158.7, 150.3,
138.5, 137.8, 137.3, 130.5, 130.2, 129.6, 129.5, 129.3, 129.1, 128,4
128.3, 128.0, 128.0, 127.5, 127.4, 127.3, 127.2, 126.8, 125.2, 125.1,
120.0, 120.0, 119.9, 119.8, 113.5, 113.4, 96.4, 94.5, 80.2, 79.9,
79.3, 79.2, 77.7, 75.2, 75.1, 74.9, 70.2, 69.7, 65.3, 55.2, 55.2; 31P
NMR (CDCl3, 121 Hz) δ -11.5; IR (film, cm-1) 3031, 2949, 1608,
1513, 1489, 1451, 1363, 1300, 1250, 1193, 1136, 1029, 960, 828;
TLC Rf 0.47 (30% ethyl acetate/hexanes); exact mass calcd for
[C57H59O13P + Na]+ requires m/z 1005.3591, found 1005.3588
(ESI+); [R]D ) +18.6 (c 1.0, CH2Cl2).
5H), 7.12 (d, J ) 8.1 Hz, 4H), 6.81 (d, J ) 8.1 Hz, 4H), 5.52 (s,
1H), 4.63, (s, 2H), 4.57-4.35 (m, 5H), 4.30 (t, J ) 3.7 Hz, 2H),
4.26 (s, 2H), 4.01 (s, 1H), 3.80 (s, 6H); 13C NMR (CDCl3, 100
MHz) δ 159.1, 137.7, 129.5, 129.2, 128.3, 127.9, 127.6, 113.7,
103.1, 73.8, 71.5, 71.2, 70.6, 68.2, 67.4, 55.3; IR (film, cm-1) 2955,
1615, 1583, 1508, 1457, 1382; TLC Rf 0.50 (50% ethyl acetate/
hexanes); exact mass calcd for [C30H32O8P + Na]+ requires m/z
543.1995, found 543.1983 (ESI+). This material (500 mg, 0.960
mmol) was then dissolved in methanol (18.0 mL) and 2 N HCl
(1.00 mL) was added. Precipitate immediately formed upon
addition, and dichloromethane was added until the precipitate
completely dissolved. The reaction was then allowed to stir at room
temperature for 18 h at which time the solution was brought to pH
) 8 with concentrated ammonium hydroxide. The reaction was
then concentrated to provide an oil containing ammonium chloride
precipitate. Ethyl acetate (200 mL) was then added, and the resulting
solution was filtered to remove ammonium chloride. The ethyl
acetate was then removed in vacuo to afford an oil. The crude
material was then purified via column chromatography eluting with
a gradient of 40-50% ethyl acetate/hexanes to afford pure triol 3
as a clear oil (271 mg, 55%). 1H NMR (CDCl3, 400 MHz) δ 7.42-
7.28 (m, 9H), 6.90 (d, J ) 8.8 Hz, 4H), 4.84 (s, 2H), 4.79 (s, 4H),
4.00 (t, J ) 2.9 Hz, 1H), 3.81 (s, 6H), 3.68-3.46 (m, 5H), 2.49 (s,
1H), 2.35 (d, J ) 5.9 Hz, 2H); 13C NMR (CDCl3, 100 MHz) δ
159.1, 138.4, 130.5, 129.6, 128.9, 128.3, 127.7, 113.9, 81.5, 79.3,
75.2, 74.9, 74.6, 72.5, 55.3; IR (film, cm-1) 3547, 2937, 1621, 1583,
1527, 1457, 1363, 1300; TLC Rf 0.13 (50% ethyl acetate/hexanes);
exact mass calcd for [C29H34O8 + Na]+ requires m/z 533.2151,
found 533.2158 (ESI+).
2-O-Benzyl-1,5-di-O-benzyloxymethyl-4,6-di-O-p-methoxy-
benzyl-D-myo-inositol-3-dibenzyl Phosphate (5). To a stirred
solution of 2-O-benzyl-3,5-di-O-benzyloxymethyl-4,6-di-O-p-meth-
oxybenzyl-D-myo-inositol-3-diphenyl phosphate (17, 290 mg, 0.295
mmol) in THF (6.00 mL) was added benzyl alcohol (93.0 µL, 0.885
mmol) followed by sodium hydride (28.0 mg, 1.18 mmol). The
reaction was stirred at room temperature for 35 min and was then
quenched with saturated NH4Cl. The THF was then removed in
vacuo and the resulting residue was extracted with ethyl acetate (3
× 100 mL). The combined organic layers were dried (MgSO4),
filtered, and concentrated in vacuo to afford an orange oil. This
crude material was then purified by column chromatography (10-
36% ethyl acetate/hexanes) to afford pure 2-O-benzyl-3,5-di-O-
benzyloxymethyl-4,6-di-O-p-methoxybenzyl-D-myo-inositol-3-diben-
1
zyl phosphate (5) as a clear thick oil (230 mg, 77%). H NMR
(CDCl3, 300 MHz) δ 7.42-7.08 (m, 29H), 6.76 (d, J ) 8.7 Hz,
2H), 6.70 (d, J ) 8.7 Hz, 2H), 5.04-4.66 (m, 14H), 4.56 (s, 2H),
4.54-4.49 (m, 2H), 4.32-4.19 (m, 3H), 3.98 (ABq, J ) 9.9 Hz,
2H), 3.75 (s, 3H), 3.71 (s, 3H), 3.57 (ABq, J ) 9.4 Hz, 2H); 13C
NMR (CDCl3, 100 MHz) δ 158.8, 138.6, 137.8, 137.4, 135.6, 135.5,
130.5, 130.3, 129.2, 129.0, 128.3, 128.3, 128.2, 128.0, 127.7, 127.6,
127.6, 127.5, 127.4, 127.3, 127.2, 113.5, 113.5, 96.4, 93.9, 80.2,
79.5, 79.4, 79.3, 78.8, 78.7, 75.2, 75.1, 75.0, 70.1, 69.4, 69.4, 69.2,
2-O-Benzyl-4,6-di-O-p-methoxybenzyl-D-myo-inositol-1-di-
phenyl Phosphate (ent-4). 2-O-Benzyl-4,6-di-O-p-methoxybenzyl-
D-myo-inositol (3) was phosphorylated under standard peptide
catalyzed conditions using Peptide 1P.4 Product was obtained in
46% yield and 97% ee. 1H NMR (CDCl3, 400 MHz) δ 7.39-7.11
(m, 19H), 6.87 (d, J ) 8.1 Hz, 2H), 6.80 (d, J ) 8.1 Hz, 2H),
4.89-4.52 (m, 7H), 4.22 (t, J ) 2.2 Hz, 1H), 3.92 (t, J ) 8.8 Hz,
1H), 3.79 (s, 3H), 3.77 (s, 3H), 3.72-3.47 (m, 3H), 2.46 (s, 1H),
2.26 (d, J ) 5.1 Hz, 1H); 13C NMR (CDCl3, 100 MHz) δ 159.0,
138.3, 130.4, 130.1, 129.6, 129.5, 128.1, 127.4, 127.2, 125.3, 125.2,
119.9, 119.8, 113.8, 113.7, 80.4, 79.8, 79.8, 79.7, 79.7, 78.9, 75.2,
74.8, 74.6, 74.5, 71.7, 55.2; 31P NMR (CDCl3, 162 Hz) δ -11.2;
IR (film, cm-1) 3427, 3056, 2936, 2829, 1608, 1598, 1508, 1489,
1250; TLC Rf 0.20 (50% ethyl acetate/hexanes); exact mass calcd
for [C41H43O11P + Na]+ requires m/z 765.2441, found 765.2439
(ESI+). HPLC tR 15.4 min employing a normal phase Chiralcel
69.2, 55.2, 55.2; 31P NMR (CDCl3, 121 Hz) δ -0.6; IR (film, cm-1
)
3031, 2936, 2892, 2357, 2332, 1608, 1514, 1457, 1376, 1250, 1168,
1124, 1023; TLC Rf 0.22 (30% ethyl acetate/hexanes); exact mass
calcd for [C59H63O13P + Na]+ requires m/z 1033.3904, found
1033.3887 (ESI+); [R]D ) +12.0 (c 1.0, CH2Cl2).
2-O-Benzyl-1,5-di-O-benzyloxymethyl-D-myo-inositol-3-diben-
zyl Phosphate (6). To a stirred solution of 2-O-benzyl-3,5-di-O-
benzyloxymethyl-4,6-di-O-p-methoxybenzyl-D-myo-inositol-3-diben-
zyl phosphate (5, 220 mg, 0.218 mmol) in a 9:1 mixture of DCM/
water (6.00 mL total) was added DDQ (148 mg, 0.653 mmol). The
reaction was stirred at room temperature for 2 h at which time the
reaction was diluted with excess DCM and saturated NaHCO3 was
added. The reaction was then extracted with DCM. The organic
layers were combined, dried (MgSO4), filtered and concentrated
in vacuo. The resulting residue was then purified by column
chromatography (12-50% ethyl acetate/hexanes) to afford pure
2-O-benzyl-3,5-di-O-benzyloxymethyl-D-myo-inositol-3-dibenzyl
OD column (Alltech), eluting with 30% ethanol/hexanes [R]D
+5.4 (c 1.0, CH2Cl2, at 97% ee).
)
2-O-Benzyl-4,6-di-O-p-methoxybenzyl-D-myo-inositol-3-di-
phenyl Phosphate (4). 2-O-Benzyl-4,6-di-O-p-methoxybenzyl-D-
myo-inositol (3) was phosphorylated under standard peptide-
catalyzed conditions using Peptide 3P.4 Product was obtained in
53% yield and 98% ee. Spectral data matched that for 2-O-benzyl-
4,6-di-O-p-methoxybenzyl-D-myo-inositol-1-diphenyl phosphate
above. HPLC tR 12.9 min employing a normal phase Chiralcel OD
column (Alltech), eluting with 30% ethanol/hexanes [R]D ) -4.2
(c 1.0, CH2Cl2, at 98% ee).
2-O-Benzyl-1,5-di-O-benzyloxymethyl-4,6-di-O-p-methoxy-
benzyl-D-myo-inositol-3-diphenyl Phosphate (17). To a stirred
solution of 2-O-benzyl-4,6-di-O-p-methoxybenzyl-D-myo-inositol-
3-diphenyl phosphate (4, 535 mg, 0.720 mmol) in DMF (15.0 mL)
was added diisopropyl ethylamine (878 µL, 5.04 mmol) followed
by BOMCl (601 µL, 4.32 mmol). The reaction was stirred at room
temperature for 18 h and then concentrated in vacuo to remove
DMF. The resulting residue was then purified by column chroma-
tography eluting with 20% ethyl acetate/hexanes to afford pure 2-O-
benzyl-3,5-di-O-benzyloxymethyl-4,6-di-O-p-methoxybenzyl-D-
myo-inositol-3-diphenyl phosphate as a clear oil (465 mg, 66%).
1H NMR (CDCl3, 300 MHz) δ 7.44-7.02 (m, 29H), 6.77 (d, J )
1
phosphate (6) as a white solid (134 mg, 80%). H NMR (CDCl3,
300 MHz) δ 7.60-7.17 (m, 25H), 5.11 (d, J ) 7.8 Hz, 2H), 5.05
(dd, J ) 5.1, 8.0 Hz, 2H), 4.88-4.51 (m, 8H), 4.23 (ddd, J ) 2.3,
7.6, 9.9 Hz, 1H), 4.16-4.01 (m, 3H), 3.77 (d, J ) 2.3 Hz, 1H),
3.54 (d, J ) 1.6 Hz, 1H), 3.48 (dd, J ) 2.3, 9.9 Hz, 1H), 3.26 (t,
J ) 9.0 Hz, 1H); 13C NMR (CDCl3, 100 MHz) δ 138.3, 137.2,
136.6, 135.7, 135.6, 135.6, 135.5, 128.4, 128.3, 128.2, 128.1, 128.0,
127.9, 127.8, 127.7, 127.7, 127.6, 127.6, 127.6, 127.6, 127.5, 127.5,
127.4, 127.3, 127.3, 96.5, 94.2, 85.5, 78.9, 78.8, 77.8, 77.4, 77.3,
75.0, 71.0, 70.9, 70.9, 70.4, 69.8, 69.7, 69.7, 69.4; 31P NMR (CDCl3,
121 Hz) δ 0.2; IR (film, cm-1) 3502, 3352, 3031, 2986, 1495, 1457,
1382, 1250, 1155, 1117, 1086, 1023; TLC Rf 0.14 (40% ethyl
acetate/hexanes); exact mass calcd for [C43H47O11P + Na]+ requires
m/z 793.2754, found 793.2779 (ESI+); [R]D ) -1.8 (c 1.0, CH2-
Cl2).
J. Org. Chem, Vol. 71, No. 18, 2006 6927

Morgan et al.
2-O-Benzyl-1,5-di-O-benzyloxymethyl-D-myo-inositol-3,4,6-
tris(dibenzyl) Phosphate (7). To a stirred solution of 2-O-benzyl-
3,5-di-O-benzyloxymethyl-D-myo-inositol-3-dibenzyl phosphate (6,
65.0 mg, 0.0840 mmol) in DCM (4.00 mL) was added dibenzyl
diisopropylphosphoramidite (141 µL, 0.420 mmol) followed by 4,5-
dicyanoimidazole (60.0 mg, 0.504 mmol). The reaction was stirred
at room temperature for 15 h at which time the reaction was cooled
to 0 °C and 1 mL 30% H2O2 was added. The reaction continued to
stir at 0 °C for 1 h at which time the reaction was quenched with
saturated Na2SO3 until bubbling ceased. The reaction was then
extracted with DCM (5 × 100 mL) and the organic layers were
combined, dried (MgSO4), filtered, and concentrated in vacuo. The
resulting residue was then purified by column chromatography (20-
40% ethyl acetate/hexanes) to afford pure 2-O-benzyl-3,5-di-O-
benzyloxymethyl-D-myo-inositol-3,4,6-tris(dibenzyl) phosphate (7)
as a thick oil (85.0 mg, 78%). 1H NMR (CDCl3, 300 MHz) δ 7.55-
7.11 (m, 45H), 5.09-4.84 (m, 15H), 4.84-4.70 (m, 2H), 4.67-
4.54 (m, 4H), 4.53-4.38 (m, 4H), 4.22 (ddd, J ) 2.1, 7.3, 9.6 Hz,
remove DMF. The resulting residue was then extracted with
dichloromethane and the organic layers were combined, dried
(MgSO4), filtered and concentrated to afford a clear thick oil. The
product was then purified by column chromatography eluting with
10% ethyl acetate/hexanes to afford pure 2-O-benzyl-4,6-di-O-p-
methoxybenzyl-1,5-di-O-tert-butyldimethylsilyl-D-myo-inositol-3-
diphenyl phosphate as a clear oil (284 mg, 90%). 1H NMR (CDCl3,
400 MHz) δ 7.40-7.05 (m, 19H), 6.82 (d, J ) 8.7 Hz, 2H), 6.75
(d, J ) 8.7 Hz, 2H), 4.94-4.84 (m, 2H), 4.76-4.63 (m, 3H), 4.58
(ddd, J ) 2.5, 7.8, 10.1 Hz, 1H), 4.48 (d, J ) 11.2 Hz, 1H), 4.12-
4.07 (m, 1H), 3.90 (t, J ) 9.2 Hz, 1H), 3.78 (s, 3H), 3.76 (s, 3H),
3.71-3.62 (m, 2H), 3.53 (t, J ) 8.7 Hz, 1H), 0.82 (s, 9H), 0.80 (s,
9H), 0.09 (s, 3H), -0.01 (s, 3H), -0.07 (s, 3H), -0.12 (s, 3H);
13C NMR (CDCl3, 100 MHz) δ 158.3, 158.0, 150.2, 138.9, 133.2,
132.1, 131.1, 130.6, 129.6, 129.4, 128.6, 128.0, 127.3, 127.2, 127.1,
125.3, 125.1, 124.9, 123.7, 119.9, 119.7, 113.1, 113.0, 81.4, 80.7,
80.2, 80.1, 79.8, 75.3, 75.0, 74.8, 74.6, 73.9, 55.1, 26.1, 26.0, 18.1,
-3.7, -4.1, -4.7; 31P NMR (CDCl3, 121 Hz) δ -11.1; IR (film,
cm-1) 3064, 3032, 3001, 2953, 2929, 2895, 2856, 1614, 1589, 1490,
1472, 1463, 1387, 1359, 1295, 1248, 1220, 1191, 1170, 1127, 1072,
1037, 957; exact mass calcd for [C53H71O11PSi2 + H]+ requires
m/z 971.4351, found 971.4376 (ESI+). [R]D ) +21.2 (c 1.0, CH2-
Cl2).
1H), 3.70 (t, J ) 9.2 Hz, 1H), 3.61 (dd, J ) 2.3, 10.1 Hz, 1H); 13
C
NMR (CDCl3, 100 MHz) δ 150.8, 143.4, 138.0, 138.0, 137.2, 135.8,
135.7, 135.6, 135.4, 135.3, 133.0, 128.6, 128.5, 128.5, 128.3, 128.2,
128.2, 128.1, 128.0, 128.0, 127.8, 127.7, 127.6, 127.5, 127.5, 127.4,
127.4, 127.4, 127.3, 127.1, 126.8, 126.8, 126.3, 126.0, 125.8, 124.8,
123.9, 120.8, 120.3, 119.7, 118.8, 113.6, 110.5, 109.2, 104.3, 101.9,
101.1, 96.1, 94.4, 91.8, 87.1, 78.4, 78.4, 77.7, 76.5, 76.4, 76.2, 75.8,
75.7, 75.1, 70.6, 69.8, 69.8, 69.6, 69.5, 69.4, 69.3, 69.3, 58.8, 58.1;
2-O-Benzyl-4,6-di-O-p-methoxybenzyl-1,5-di-O-tert-butyldi-
methylsilyl-D-myo-inositol-3-dibenzyl Phosphate (8). To a stirred
solution of 2-O-benzyl-4,6-di-O-p-methoxybenzyl-1,5-di-O-tert-
butyl dimethylsilyl-D-myo-inositol-3-diphenyl phosphate (18, 280
mg, 0.288 mmol) in THF (5.80 mL) was added benzyl alcohol (89.0
µL, 0.864 mmol) followed by sodium hydride (27.6 mg, 1.15
mmol). The reaction was stirred at room temperature for 1 h at
which time saturated ammonium chloride solution was added slowly
to quench any remaining sodium hydride. The reaction was then
extracted (3 × 200 mL EtOAc) and the organic layers were
combined, dried (MgSO4), filtered and concentrated in vacuo. The
resulting residue was then purified by column chromatography
eluting with 15% ethyl acetate/hexanes to afford pure 2-O-benzyl-
4,6-di-O-p-methoxybenzyl-1,5-di-O-tert-butyldimethylsilyl-D-myo-
inositol-3-dibenzyl phosphate (8) as a clear oil (254 mg, 88%). 1H
NMR (CDCl3, 400 MHz) δ 7.40-7.12 (m, 19H), 7.82 (d, J ) 8.7
Hz, 2H), 6.75 (d, J ) 8.5 Hz, 2H), 5.00-4.80 (m, 6H), 4.79-4.63
(m, 4H), 4.24 (ddd, J ) 2.5, 7.6, 10.1 Hz, 1H), 4.13 (br s, 1H),
3.85 (t, J ) 9.2 Hz, 1H), 3.79 (s, 3H), 3.72 (s, 3H), 3.68 (d, J )
9.2 Hz, 1H), 3.57 (dd, J ) 2.3, 9.6 Hz, 1H), 3.46 (t, J ) 9.0 Hz,
31P NMR (CDCl3, 121 Hz) δ -0.2, -0.4, -0.8; IR (film, cm-1
)
3064, 3032, 2956, 2924, 1497, 1455, 1380, 1275, 1214, 1023; exact
mass calcd for [C71H73O17P3 + H]+ requires m/z 1291.4139, found
1291.4186 (ESI+); [R]D ) +14.3 (c 1.0, CH2Cl2).
D-myo-Inositol-3,4,6-triphosphate Hexasodium Salt (D-I-3,4,-
6P3). To a stirred solution of 2-O-benzyl-3,5-di-O-benzyloxymethyl-
D-myo-inositol-3,4,6-tris(dibenzyl) phosphate (7, 40.0 mg, 0.031
mmol) in methanol (2.50 mL) was added 20% palladium hydroxide
on carbon (30.0 mg). The reaction was sealed and then purged with
hydrogen several times. The reaction continued to stir at room
temperature under hydrogen atmosphere (balloon pressure) for 24
h. The reaction was then filtered through Celite into a round-bottom
flask and prerinsed (3 × H2O) Chelex 100 resin (sodium form)
was added to afford the crude product as the hexasodium salt. Minor
impurities were removed by converting this material to the free
acid form via treatment with Dowex 50X2-200 followed by
conversion back to the hexasodium salt as described above. The
light brown solid obtained upon lyophilization was then washed
several times with methanol to afford pure D-myo-inositol-3,4,6-
triphosphate hexasodium salt (D-I-3,4,6P3) (15 mg, 88%). 1H NMR
(D2O, pD 10, 300 MHz) δ 4.34 (br. s, 1H), 4.26-4.09 (m, 2H),
3.89 (ddd, J ) 2.5, 7.6, 9.9 Hz, 1H), 3.68 (dd, J ) 3.0, 9.6 Hz,
1H), 3.53 (t, J ) 9.0 Hz, 1H); 13C NMR (D2O, pD 10, 126 MHz)
δ 76.9 (d, J ) 5.4 Hz, 1C), 75.9 (t, J ) 6.0 Hz, 1C), 75.0 (d, J )
6.3 Hz, 1C), 74.0 (t, J ) 5.5 Hz, 1C), 71.6 (s, 1C), 70.9 (s, 1C);
31P NMR (D2O, pD 10, 121 Hz) δ 6.0, 5.5, 4.4; exact mass calcd
for [C6H15O15P3 - H]- requires m/z 418.9546, found 418.9530
(ESI-). [R]D ) +5.3 (c 0.8, H2O, pH 8) [lit. (D-I-1,4,6P4) [R]D )
-10.1 (c 0.78, H2O),12 [R]D ) -8.9 (c 0.90, H2O, sodium salt).13
2-O-Benzyl-4,6-di-O-p-methoxybenzyl-1,5-di-O-tert-butyldi-
methylsilyl-D-myo-inositol-3-diphenyl Phosphate (18). To a stirred
solution of 2-O-benzyl-4,6-di-O-p-methoxybenzyl-D-myo-inositol-
3-diphenyl phosphate (4, 240 mg, 0.323 mmol) in DMF (4.50 mL)
was added imidazole (440 mg, 6.46 mmol) followed by tert-butyl
dimethylsilyl chloride (487 mg, 3.23 mmol). The reaction was
stirred at 50 °C for 48 h at which time the reaction was quenched
with methanol and continued to stir for 30 min. The reaction was
then cooled to room temperature and concentrated in vacuo to
1H), 0.81 (s, 18H), 0.03 (s, 3H), -0.03 (s, 6H), -0.11 (s, 3H); 13
C
NMR (CDCl3, 100 MHz) δ 158.5, 158.0, 139.1, 131.2, 130.7, 128.8,
128.4, 128.3, 128.3, 128.0, 127.7, 127.6, 127.3, 127.2, 127.1, 126.8,
113.2, 113.0, 81.5, 80.7, 80.1, 80.0, 78.9, 78.9, 75.5, 75.0, 74.9,
74.6, 73.9, 69.3, 69.2, 69.1, 55.2, 26.1, 26.0, 18.1, 18.0, -3.7, -4.2;
31P NMR (CDCl3, 162 Hz) δ -0.4; IR (film, cm-1) 3065, 3033,
2953, 2929, 2895, 2856, 1614, 1514, 1463, 1386, 1248, 1127, 1076,
1035, 1016; exact mass calcd for [C55H75O11PSi2 + H]+ requires
m/z 999.4664, found 999.4689 (ESI+). [R]D ) +19.4 (c 1.0, CH2-
Cl2).
2-O-Benzyl-4,6-di-O-p-methoxybenzyl-1-O-tert-butyldimeth-
ylsilyl-D-myo-inositol-3-dibenzyl Phosphate (19). To a stirred
solution of 2-O-benzyl-4,6-di-O-p-methoxybenzyl-D-myo-inositol-
3-diphenyl phosphate (8, 1.44 g, 1.44 mmol) in methanol (29.0
mL) was added acetyl chloride (10.0 µL, 0.144 mmol). The reaction
was stirred at 0 °C for 12 h at which time saturated aqueous sodium
bicarbonate was added. The reaction was then concentrated in vacuo
and extracted (5 × 200 mL DCM). The organic layers were
combined, dried (MgSO4), filtered and concentrated in vacuo to
afford a clear oil. This crude material was then purified by column
chromatography eluting with a gradient of 20-35% ethyl acetate/
hexanes to afford pure 2-O-benzyl-4,6-di-O-p-methoxybenzyl-1-
O-tert-butyldimethylsilyl-D-myo-inositol-3-dibenzyl phosphate as a
(12) Chung, S.; Kwon, Y.; Shin, J.; Chang, Y.; Lee, C.; Shin, B.; Kim,
K.; Kim, M. J. Org. Chem. 2002, 67, 5626-5637.
(13) Watanabe, Y.; Ogasawara, T.; Ozaki, S.; Hirata, M. Carbohydr.
Res. 1994, 258, 87-92.
1
clear oil (630 mg, 49%). H NMR (CDCl3, 400 MHz) δ 7.40-
7.18 (m, 19H), 6.86 (d, J ) 8.5 Hz, 2H), 6.76 (d, J ) 8.3 Hz, 2H),
5.10-4.60 (m, 10H), 4.23 (ddd, J ) 2.5, 7.8, 9.9 Hz, 1H), 4.09
6928 J. Org. Chem., Vol. 71, No. 18, 2006

Unified Syntheses of IPn’s
(br. s, 1H), 3.89 (t, J ) 9.4 Hz, 1H), 3.80-3.68 (m, 7H), 3.56 (dd,
J ) 1.8, 9.4 Hz, 1H), 3.43 (t, J ) 9.2 Hz, 1H), 2.34 (br. s, 1H),
0.92 (s, 9H), 0.09 (s, 3H), 0.06 (s, 3H); 13C NMR (CDCl3, 100
MHz) δ 158.9, 158.9, 138.8, 135.6, 130.8, 130.5, 129.4, 129.2,
128.3, 128.3, 128.2, 128.2, 128.0, 127.7, 127.6, 127.1, 127.0, 113.7,
113.5, 81.0, 80.9, 79.5, 79.4, 78.3, 78.2, 75.5, 75.0, 74.8, 74.6, 73.5,
69.3, 69.3, 69.1, 69.0, 55.2, 55.2, 26.0, 18.1, -4.4; 31P NMR
(CDCl3, 121 Hz) δ -0.5; IR (film, cm-1) 3428, 2954, 2927, 2855,
1652, 1636, 1615, 1514, 1457, 1249, 1117, 1021; exact mass calcd
for [C49H61O11PSi + Na]+ requires m/z 907.3618, found 907.3615
(ESI+). [R]D ) +28.4 (c 1.0, CH2Cl2).
acetate/hexanes); exact mass calcd for [C47H58O12P2 + Na]+ requires
m/z 927.3071, found 927.3057 (ESI+). [R]D ) +6.2 (c 1.0, CH2-
Cl2).
2-O-Benzyl-1-O-tert-butyldimethylsilyl-D-myo-inositol-3,4,5-
tris(dibenzyl) Phosphate (11). To a stirred solution of 2-O-benzyl-
1-O-tert-butyl dimethylsilyl-D-myo-inositol-3,5-bis(dibenzyl) phos-
phate (10, 50.0 mg, 0.0550 mmol) in DCM (2.50 mL) was added
dibenzyl diisopropylphosphoramidite (28.0 µL, 0.083 mmol) fol-
lowed by 4,5-dicyanoimidazole (13.0 mg, 0.110 mmol). The
reaction was stirred at room temperature for 11 h at which time
starting material was still present, so another 10.0 µL dibenzyl
diisopropyl phosphoramidite was added and the reaction continued
to stir for 4 h. The reaction was then cooled to 0 °C, 30% H2O2
(600 µL) was added, and the mixture stirred for 1 h. The reaction
was then quenched with saturated aqueous Na2SO3 until bubbling
ceased. The mixture was then extracted with DCM (5 × 50 mL)
and the organic layers were combined, dried (MgSO4), filtered and
concentrated in vacuo. The resulting residue was then purified by
column chromatography (15-35% ethyl acetate/hexanes) to afford
pure 2-O-benzyl-1-O-tert-butyldimethylsilyl-D-myo-inositol-3,4,5-
2-O-Benzyl-4,6-di-O-p-methoxybenzyl-1-O-tert-butyldimethyl-
silyl-D-myo-inositol-3,5-bis(dibenzyl) Phosphate (9). To a stirred
solution of 2-O-benzyl-4,6-di-O-p-methoxybenzyl-1-O-tert-butyl
dimethylsilyl-D-myo-inositol-3-dibenzyl phosphate (19, 530 mg,
0.599 mmol) in DCM (29.5 mL) was added dibenzyl diisopropyl
phosphoramidite (1.00 mL, 3.00 mmol) followed by 4,5-dicy-
anoimidazole (424 mg, 3.59 mmol). The reaction was stirred at
room temperature for 15 h at which time the reaction was cooled
to 0 °C and 3 mL of 30% H2O2 was added. The reaction continued
to stir at 0 °C for 1 h at which time the reaction was quenched
slowly with saturated aqueous Na2SO3 until bubbling ceased. The
reaction was then extracted with DCM (5 × 100 mL) and the
organic layers were combined, dried (MgSO4), filtered, and
concentrated. The resulting residue was then purified by column
chromatography (15-30% ethyl acetate/hexanes) to afford pure
2-O-benzyl-4,6-di-O-p-methoxybenzyl-1-O-tert-butyldimethylsilyl-
D-myo-inositol-3,5-bis(dibenzyl) phosphate (9) as a thick oil (600
1
tris(dibenzyl) phosphate (11) as a white solid (45.0 mg, 70%). H
NMR (CDCl3, 300 MHz) δ 7.40-7.07 (m, 35H), 5.20-4.82 (m,
14H), 4.64 (d, J ) 11.5 Hz, 1H), 4.37 (d, J ) 2.3 Hz, 1H), 4.27-
4.15 (m, 2H), 4.14-3.96 (m, 2H), 3.49 (br. d, J ) 7.1 Hz, 1H),
0.89 (s, 9H), 0.13 (s, 3H), 0.08 (s, 3H); 13C NMR (CDCl3, 100
MHz) δ 138.5, 135.9, 135.8, 135.7, 135.6, 135.5, 135.5, 135.4,
135.4, 135.4, 135.3, 135.3, 128.4, 128.4, 128.3, 128.2, 128.0, 128.0,
128.0, 127.9, 127.8, 127.7, 127.6, 127.2, 127.2, 81.8, 79.6, 77.2,
75.8, 75.7, 73.1, 72.3, 70.1, 70.0, 69.8, 69.7, 69.5, 69.4, 69.3, 69.3,
1
69.2, 32.0, 30.2, 29.8, 29.4, 26.0, 22.8, 18.4, 14.2, -4.1, -4.7; 31
P
mg, 87%). H NMR (CDCl3, 300 MHz) δ 7.40-7.09 (m, 25H),
NMR (CDCl3, 121 Hz) δ 0.9, -0.4, -1.0; IR (film, cm-1) 3356,
2954, 2924, 2853, 1659, 1632, 1456, 1377, 1271, 1012; TLC Rf
0.50 (50% ethyl acetate/hexanes); exact mass calcd for [C61H71O15P3-
Si + Na]+ requires m/z 1187.3673, found 1187.3705 (ESI+). [R]D
) +11.6 (c 1.0, CH2Cl2).
7.07-6.93 (m, 4H), 6.77 (d, J ) 9.0 Hz, 2H), 6.70 (d, J ) 8.7 Hz,
2H), 5.00-4.57 (m, 14H), 4.40 (ABq, J ) 9.2 Hz, 1H), 4.25 (ddd,
J ) 2.5, 7.6, 9.9 Hz, 1H), 4.13 (br. s, 1H), 4.05 (t, J ) 9.5 Hz,
1H), 3.90 (t, J ) 9.4 Hz, 1H), 3.74 (s, 3H), 3.69 (s, 3H), 3.59 (dd,
J ) 2.1, 9.4 Hz, 1H), 0.85 (s, 9H), 0.01 (s, 3H), -0.03 (s, 3H); 13
C
NMR (CDCl3, 100 MHz) δ 158.6, 158.3, 138.7, 135.9, 135.8, 135.4,
130.6, 130.2, 129.0, 128.3, 128.2, 128.1, 128.0, 128.0, 127.8, 127.7,
127.6, 127.4, 127.2, 127.0, 113.3, 113.2, 80.6, 80.3, 79.3, 78.1,
78.0, 77.9, 75.5, 74.4, 74.1, 73.4, 69.4, 69.2, 69.1, 69.0, 69.0, 68.9,
55.1, 25.9, 18.0, -4.4, -4.7; 31P NMR (CDCl3, 121 Hz) δ -0.3,
-0.6; IR (film, cm-1) 3064, 3033, 2953, 2929, 2855, 1679, 1613,
1586, 1514, 1498, 1456, 1381, 1360, 1249, 1214, 1174, 1157, 1129,
1082, 1011; TLC Rf 0.15 (30% ethyl acetate/hexanes); exact mass
calcd for [C63H74O14P2Si + H]+ requires m/z 1145.4401, found
1145.4454 (ESI+). [R]D ) +9.7 (c 1.0, CH2Cl2).
2-O-Benzyl-1-O-tert-butyldimethylsilyl-D-myo-inositol-3,5-bis-
(dibenzyl) Phosphate (10). To a stirred solution of 2-O-benzyl-
4,6-di-O-p-methoxybenzyl-1-O-tert-butyl dimethylsilyl-D-myo-
inositol-3,5-bis(dibenzyl) phosphate (9, 300 mg, 0.262 mmol) in a
9:1 mixture of DCM/water (7.00 mL) was added DDQ (178 mg,
0.786 mmol). The reaction was stirred at room temperature for 12
h and then diluted with excess DCM (100 mL). The reaction was
then washed several times with saturated aqueous NaHCO3 until
the organic layer became clear. The organic layer was then dried
(MgSO4), filtered, and concentrated in vacuo. The resulting residue
was then purified by column chromatography eluting with 40%
ethyl acetate/hexanes to afford pure 2-O-benzyl-1-O-tert-butyl
dimethylsilyl-D-myo-inositol-3,5-bis(dibenzyl) phosphate (10) as a
white solid (158 mg, 67%). 1H NMR (CDCl3, 300 MHz) δ 7.40-
7.20 (m, 25H), 5.14-4.93 (m, 8H), 4.71 (ABq, J ) 11.2 Hz, 2H),
4.30-4.03 (m, 3H), 4.01-3.81 (m, 3H), 3.49 (d, J ) 9.2 Hz, 1H),
2.98 (d, J ) 2.5 Hz, 1H), 0.90 (s, 9H), 0.11 (s, 3H), 0.07 (s, 3H);
13C NMR (CDCl3, 100 MHz) δ 138.5, 135.6, 135.5, 128.4, 128.4,
128.3, 128.0, 127.9, 127.8, 127.7, 127.2, 127.1, 82.8, 82.7, 80.2,
78.3, 75.5, 73.3, 72.0, 72.1, 71.0, 71.0, 69.7, 69.8, 69.4, 69.4, 29.8,
25.9, 18.3, -4.4, -4.5; 31P NMR (CDCl3, 121 Hz) δ 0.9, 0.1; IR
(film, cm-1) 3390, 3091, 3065, 3034, 2953, 2928, 2897, 2855, 1498,
1456, 1386, 1360, 1260, 1216, 1157, 1016; TLC Rf 0.54 (60% ethyl
D-myo-Inositol-3,4,5-triphosphate Hexasodium Salt (D-I-3,4,-
5P3). To a stirred solution of 2-O-benzyl-1-O-tert-butyl dimethyl-
silyl-D-myo-inositol-3,4,5-tris(dibenzyl) phosphate (11, 37 mg, 0.032
mmol) in HPLC grade methanol (1.0 mL) was added 20%
palladium hydroxide on carbon (30 mg). The mixture was stirred
under hydrogen (atmospheric pressure) for 24 h and then filtered
through a syringe filter (Millex-HV 0.45 µm) to remove the catalyst.
Solvent was then removed under reduced pressure to afford D-I-
3,5,6P3 in free acid form. This material was redissolved in H2O
and Chelex 100 (sodium form) was added and the mixture stirred
for 3 min then filtered. The solvent was then removed via
lyophilization and the resulting solid was rinsed several times with
HPLC grade methanol to afford D-I-3,4,5P3 as the hexasodium salt
1
(13 mg, 97%). H NMR (D2O, sodium salt, pD ∼8, 300 MHz) δ
4.32 (q, J ) 9.4 Hz, 1H), 4.22 (t, J ) 3.0 Hz, 1H), 3.99 (td, J )
2.8, 9.4 Hz, 1H), 3.90 (q, J ) 8.7 Hz, 1H), 3.81 (t, J ) 9.4 Hz,
1H), 3.61 (dd, J ) 2.8, 9.6 Hz, 1H); 13C NMR (D2O, pD 10, 100
MHz) δ 78.9, 77.3, 74.9, 74.9, 74.6, 73.6, 73.5, 71.8, 70.3, 70.2;
31P NMR (D2O, sodium salt, pD ∼8, 121 Hz) δ 4.8, 3.8, 2.1; exact
mass calcd for [C6H15O15P3 + H]+ requires m/z 420.9702, found
420.9696 (ESI+). [R]D ) +0.4 (c 2.4, H2O, pH 9) [lit. (D-I-1,5,-
6P4) [R]D ) -2.57 (c 1.01, H2O),11 [R]D ) +2.2 (c 2.30, H2O,
free acid),14 [R]D ) -2.8 (c 1.43, H2O, sodium salt)15].
2-O-Benzyl-1-O-tert-butyldimethylsilyl-D-myo-inositol-3-diben-
zyl Phosphate (12). To a stirred solution of 2-O-benzyl-4,6-di-O-
p-methoxybenzyl-1-O-tert-butyldimethylsilyl-D-myo-inositol-3-diben-
zyl phosphate (19, 220 mg, 0.249 mmol) in a 9:1 mixture of DCM/
water (9.00 mL) was added DDQ (170 mg, 0.747 mmol). The
reaction was stirred at room temperature for 1.5 h and then diluted
(14) Adelt, S.; Plattenburg, O.; Stricker, R.; Reiser, G.; Altenbach, H.
J.; Vogel, G. J. Med. Chem. 1999, 42, 1262-1273.
(15) Salamonczyk, G. M.; Pietrusiewicz, K. M. Tetrahedron Lett. 1994,
35, 4233-4234.
J. Org. Chem, Vol. 71, No. 18, 2006 6929

Morgan et al.
with excess DCM (100 mL). The reaction was then washed several
times with saturated aqueous NaHCO3 until the organic layer
became clear. The organic layer was then dried (MgSO4), filtered,
and concentrated in vacuo. The resulting residue was then purified
by column chromatography (50-80% ethyl acetate/hexanes) to
afford pure 2-O-benzyl-1-O-tert-butyldimethylsilyl-D-myo-inositol-
3-dibenzyl phosphate (12) as a white solid (140 mg, 87%). 1H NMR
(CDCl3, 300 MHz) δ 7.44-7.10 (m, 15H), 5.10-4.85 (m, 4H),
4.81 (d, J ) 11.7 Hz, 1H), 4.60-4.35 (m, 3H), 4.26 (ddd, J ) 2.3,
7.8, 10.1 Hz, 1H), 4.00 (t, J ) 9.0 Hz, 1H), 3.92-3.276 (m, 2H),
3.56-3.27 (m, 3H), 0.88 (s, 9H), 0.10 (s, 3H), 0.04 (s, 3H); 13C
NMR (CDCl3, 100 MHz) δ 138.6, 135.4, 135.4, 128.3, 128.2, 128.2,
127.8, 127.8, 127.6, 127.6, 127.5, 126.9, 80.6, 78.7, 78.6, 75.3,
74.6, 73.5, 72.8, 71.9, 69.7, 69.3, 25.9, 18.2, -4.2, -4.4; 31P NMR
(CDCl3, 162 Hz) δ 0.0; IR (film, cm-1) 3427, 3089, 3065, 3033,
2953, 2927, 2894, 2854, 1497, 1456, 1423, 1386, 1335, 1249, 1220,
1159, 1137, 1098, 1077, 1041, 1014, 851, 836; TLC Rf 0.18 (80%
ethyl acetate/hexanes); exact mass calcd for [C33H45O9PSi + H]+
requires m/z 645.2649, found 645.2641 (ESI+). [R]D ) +10.5 (c
2.0, CH2Cl2).
2-O-Benzyl-1-O-benzyloxymethyl-4,6-di-O-p-methoxybenzyl-
D-myo-inositol-3-diphenyl Phosphate (21). To a stirred solution
of 2-O-benzyl-4,6-di-O-p-methoxybenzyl-D-myo-inositol-3-diphenyl
phosphate (4, 285 mg, 0.384 mmol) in THF (5.00 mL) was added
diisopropyl ethylamine (401 µL, 2.30 mmol) followed by benzy-
loxymethyl chloride (BOMCl, 267 µL, 1.92 mmol). The reaction
was stirred at 50 °C for 24 h and was then quenched with excess
methanol and concentrated. The mixture was then extracted with
DCM (5 × 100 mL) and the organic layers were combined, dried
(MgSO4), filtered and concentrated in vacuo. The resulting residue
was then purified by column chromatography (10-30% ethyl
acetate/hexanes) to afford pure 2-O-benzyl-1-O-benzyloxymethyl-
4,6-di-O-p-methoxybenzyl-D-myo-inositol-3-diphenyl phosphate as
1
a clear oil (120 mg, 36%). H NMR (CDCl3, 300 MHz) δ 7.40-
7.06 (m, 24H), 6.84 (d, J ) 8.71 Hz, 2H), 6.78 (d, J ) 8.5 Hz,
2H), 4.89-4.48 (m, 11H), 4.26 (t, J ) 2.3 Hz, 1H), 3.95 (t, J )
9.4 Hz, 1H), 3.87 (t, J ) 9.6 Hz, 1H), 3.77 (s, 3H), 3.75 (s, 3H),
3.67 (dd, J ) 2.1, 9.6 Hz, 1H), 3.53 (dt, J ) 1.6, 9.0 Hz, 1H), 2.38
(d, J ) 2.1 Hz, 1H); 13C NMR (CDCl3, 100 MHz) δ 159.0, 158.9,
150.4, 150.3, 150.2, 148.1, 142.2, 139.0, 138.5, 137.3, 132.2, 130.6,
130.3, 129.7, 129.6, 129.4, 129.3, 129.3, 128.2, 128.0, 127.6, 127.5,
127.3, 127.2, 125.2, 125.1, 119.9, 119.9, 115.1, 113.7, 113.6, 105.2,
104.5, 104.3, 99.1, 94.3, 80.1, 79.8, 79.7, 79.4, 79.3, 78.0, 77.6,
77.3, 75.0, 74.9, 74.7, 71.3, 69.7, 68.5, 67.6, 57.9, 55.3, 52.4; 31P
NMR (CDCl3, 121 Hz) δ -11.5; IR (film, cm-1) 3451, 3063, 3031,
2936, 2904, 2836, 1612, 1588, 1514, 1488, 1454, 1294, 1249, 1214,
1189, 1115, 1072, 1035, 1026, 958; TLC Rf 0.61 (50% ethyl acetate/
hexanes); exact mass calcd for [C49H51O12P + Na]+ requires m/z
885.3016, found 885.3008 (ESI+). [R]D ) +12.6 (c 1.0, CH2Cl2).
2-O-Benzyl-1-O-tert-butyldimethylsilyl-D-myo-inositol-3,4,5,6-
tetrakis(dibenzyl) Phosphate (20). To a stirred solution of 2-O-
benzyl-1-O-tert-butyldimethylsilyl-D-myo-inositol-3-dibenzyl phos-
phate (12, 100 mg, 0.155 mmol) in DCM (7.00 mL) was added
dibenzyl diisopropylphosphoramidite (521 µL, 1.55 mmol) followed
by 4,5-dicyanoimidazole (201 mg, 1.71 mmol). The reaction was
stirred at room temperature for 14 h. The reaction was then cooled
to 0 °C, 30% H2O2 (1.20 mL) was added, and the mixture stirred
for 1 h. The reaction was then quenched with saturated aqueous
Na2SO3 until bubbling ceased. The mixture was then extracted with
DCM (5 × 50 mL) and the organic layers were combined, dried
(MgSO4), filtered and concentrated in vacuo. The resulting residue
was then purified by column chromatography (20-35% ethyl
acetate/hexanes) to afford pure 2-O-benzyl-1-O-tert-butyl dimeth-
ylsilyl-D-myo-inositol-3,4,5,6-tetrakis(dibenzyl) phosphate as a clear
2-O-Benzyl-1-O-benzyloxymethyl-4,6-di-O-p-methoxybenzyl-
D-myo-inositol-3-diphenyl Phosphate-5-dibenzyl Phosphate (13).
To a stirred solution of 2-O-benzyl-1-O-benzyloxymethyl-4,6-di-
O-p-methoxybenzyl-D-myo-inositol-3-diphenyl phosphate (21, 325
mg, 0.377 mmol) in DCM (18.9 mL) was added dibenzyl
diisopropylphosphoramidite (633 µL, 1.89 mmol) followed by 4,5-
dicyanoimidazole (267 mg, 1.26 mmol). The reaction was stirred
at room temperature for 15 h. The reaction was then cooled to 0
°C, 30% H2O2 (3.00 mL) was added, and the mixture stirred for 1
h. The reaction was then quenched with saturated aqueous Na2SO3
until bubbling ceased. The mixture was then extracted with DCM
(5 × 100 mL) and the organic layers were combined, dried
(MgSO4), filtered and concentrated in vacuo. The resulting residue
was then purified by column chromatography (15-33% ethyl
acetate/hexanes) to afford pure 2-O-benzyl-1-O-benzyloxymethyl-
4,6-di-O-p-methoxybenzyl-D-myo-inositol-3-diphenyl phosphate-5-
1
thick oil (199 mg, 90%). H NMR (CDCl3, 300 MHz) δ 7.40-
7.04 (m, 45H), 4.15-4.75 (m, 19H), 4.69 (d, J ) 11.5 Hz, 1H),
4.41-4.17 (m, 3H), 3.64 (d, J ) 9.4 Hz, 1H), 0.85 (s, 9H), 0.07
(s, 3H), 0.04 (s, 3H); 13C NMR (CDCl3, 100 MHz) δ 138.1, 136.0,
135.9, 135.9, 132.5, 129.0, 129.0, 128.9, 128.8, 128.8, 128.6, 128.6,
128.3, 128.1, 127.9, 127.9, 127.8, 127.8, 127.7, 127.6, 127.2, 127.1,
126.6, 122.2, 116.4, 102.5, 101.4, 88.6, 84.2, 79.0, 76.3, 76.2, 76.0,
75.8, 75.7, 75.6, 75.5, 75.4, 71.8, 69.7, 69.6, 69.5, 69.5, 69.4, 69.3,
69.1, 44.2, 26.0, 16.0, -2.6, -4.5, -4.6; 31P NMR (CDCl3, 121
Hz) δ 0.3, 0.1, -0.3, -0.8; IR (film, cm-1) 3064, 3032, 2954, 2927,
2895, 2855, 1497, 1456, 1382, 1278, 1215, 1128, 1017, 959, 880,
839; TLC Rf 0.21 (40% ethyl acetate/hexanes); exact mass calcd
for [C75H84O18P4 + H]+ requires m/z 1425.4456, found 1425.4678.
(ESI+). [R]D ) +7.2 (c 1.0, CH2Cl2).
1
dibenzyl phosphate (13) as a clear oil (343 mg, 81%). H NMR
(CDCl3, 400 MHz) δ 7.45-7.03 (m, 34H), 6.76 (d, J ) 8.4 Hz,
2H), 6.67 (d, J ) 8.4 Hz, 2H), 4.95-4.52 (m, 13H), 4.50 (s, 2H),
4.42 (q, J ) 9.5 Hz, 1H), 4.31 (t, J ) 2.5 Hz, 1H), 4.13 (t, J ) 9.5
Hz, 1H), 4.03 (t, J ) 9.5 Hz, 1H), 3.74 (s, 3H), 3.71 (s, 3H),. 3.68
(dd, J ) 2.2, 10.3 Hz, 1H); 13C NMR (CDCl3, 100 MHz) δ 158.7,
158.6, 150.2, 150.1, 138.3, 137.2, 135.9, 135.9, 135.8, 130.4, 130.0,
129.5, 129.3, 129.0, 128.2, 128.1, 128.0, 127.9, 127.5, 127.3, 127.2,
125.2, 125.1, 120.0, 119.9, 119.8, 113.4, 113.2, 94.4, 80.6, 80.5,
79.3, 79.2, 78.7, 77.8, 77.8, 77.7, 77.5, 77.2, 76.8, 74.9, 74.7, 74.3,
69.7, 69.2, 55.2; 31P NMR (CDCl3, 121 Hz) δ -0.5, -11.7; IR
(film, cm-1) 2956, 2925, 2856, 1611, 1588, 1515, 1490, 1457, 1249,
1191, 1023, 960; TLC Rf 0.65 (50% ethyl acetate/hexanes); exact
mass calcd for [C63H64O15P2 + H]+ requires m/z 1123.3799, found
1123.3848 (ESI+). [R]D ) +8.0 (c 1.0, CH2Cl2).
D-myo-Inositol-3,4,5,6-tetraphosphate Octasodium Salt (D-I-
3,4,5,6P4). To a stirred solution of 2-O-benzyl-1-O-tert-butyl
dimethylsilyl-D-myo-inositol-3,4,5,6-tetrakis(dibenzyl) phosphate
(20, 40 mg, 0.028 mmol) in HPLC grade methanol (2.0 mL) was
added 20% palladium hydroxide on carbon (30 mg). The mixture
was stirred under hydrogen (atmospheric pressure) for 24 h and
then filtered through Celite to remove the catalyst. Chelex 100
(sodium form) was added and the mixture stirred for 3 min and
then filtered. The solvent was then removed via lyophilization and
the resulting solid was rinsed several times with HPLC grade
methanol to afford D-I-3,4,5,6P4 as the octasodium salt (15.0 mg,
1
79%). H NMR (D2O, sodium salt, pD ∼8, 300 MHz) δ 4.47 (q,
2-O-Benzyl-1-O-benzyloxymethyl-4,6-di-O-p-methoxybenzyl-
D-myo-inositol-3,5-bis(dibenzyl) Phosphate (14). To a stirred
solution of 2-O-benzyl-1-O-benzyloxymethyl-4,6-di-O-p-methoxy-
J ) 9.6 Hz, 1H), 4.34 (q, J ) 9.2 Hz, 1H), 4.25 (t, J ) 2.3 Hz,
1H), 4.22-4.05 (m, 2H), 3.75 (dd, J ) 2.8, 9.6 Hz, 1H); 13C NMR
(D2O, pD 10, 100 MHz) δ 74.8, 74.7, 73.6, 72.4, 65.1; 31P NMR
(D2O, pD 10, 121 Hz) δ 5.3, 4.8, 4.7, 4.4; exact mass calcd for
[C6H14O18P4 + H]+ requires m/z 498.9209, found 498.9235 (ESI+).
[R]D ) +2.0 (c 2.0, H2O, pH 10) [lit. (D-I-3,4,5,6P4) [R]D ) +10.1
(c 2.23, H2O, pH 8.9),11 [R]D ) +9.8 (c 1.43, H2O, pH 11.1),16
[R]D ) -6.2 (c 2.15, H2O, sodium salt)17].
(16) pH adjusted by addition of cyclohexylamine: Ozaki, S.; Ling, L.;
Ogasawara, T.; Watanabe, Y.; Hirata, M. Carbohydr. Res. 1994, 259, 307-
310.
(17) Pietrusiewicz, K. M.; Salamonczyk, G. M.; Bruzik, K. S.; Wieczorek,
W. Tetrahedron 1992, 48, 5523-5542.
6930 J. Org. Chem., Vol. 71, No. 18, 2006

Unified Syntheses of IPn’s
benzyl-D-myo-inositol-3-diphenyl phosphate-5-dibenzyl phosphate
(13, 343 mg, 0.305 mmol) in THF (6.10 mL) was added benzyl
alcohol (95.0 µL, 0.915 mmol) followed by sodium hydride (29.0
mg, 1.22 mmol). The reaction was stirred at room temperature for
30 min at which time saturated ammonium chloride solution was
added slowly to quench any remaining sodium hydride. The reaction
was then extracted (3 × 100 mL EtOAc) and the organic layers
were combined, dried (MgSO4), filtered and concentrated in vacuo.
The resulting residue was then purified by column chromatography
(15-35% ethyl acetate/hexanes) to afford pure 2-O-benzyl-1-O-
benzyloxymethyl-4,6-di-O-p-methoxybenzyl-D-myo-inositol-3,5-bis-
diisopropyl phosphoramidite (28 µL, 0.083 mmol) followed by 4,5-
dicyanoimidazole (13 mg, 0.11 mmol). The reaction was stirred at
room temperature for 12 h and then was cooled to 0 °C and 0.20
mL 30% H2O2 solution was added. The reaction then continued to
stir at 0 °C for 1 h and was then quenched with saturated Na2SO3
solution. The product was then extracted into DCM (5 × 30 mL),
and the combined organic layers were dried (MgSO4), filtered, and
concentrated in vacuo. The resulting residue was purified by column
chromatography eluting with 60% ethyl acetate/hexanes to afford
pure 2-O-benzyl-1-O-benzyloxymethyl-D-myo-inositol-3,5,6-tris-
(dibenzyl) phosphate (16) as a colorless oil that crystallized upon
standing (35 mg, 53%). 1H NMR (CDCl3, 300 MHz) δ 7.33-7.17
(m, 40H), 5.19-4.87 (m, 12H), 4.74 (ABq, J ) 11.9 Hz, 2H), 4.57
(ABq, J ) 7.1 Hz, 2H), 4.42 (q, J ) 12.2 Hz, 2H), 4.32-4.22 (m,
2H), 4.21-4.07 (m, 2H), 3.60 (dd, J ) 2.2, 10.1 Hz, 1H); 13C NMR
(CDCl3, 100 MHz) δ 138.0, 137.2, 135.7, 135.7, 135.6, 135.5,
135.3, 135.2, 128.3, 128.2, 128.0, 127.9, 127.7, 127.4, 94.4, 77.8,
77.7, 77.1, 75.2, 75.1, 71.0, 70.3, 70.2, 70.2, 70.1, 69.8, 69.7, 69.6,
1
(dibenzyl) phosphate (14) as a clear oil (135 mg, 38%). H NMR
(CDCl3, 400 MHz) δ 7.40-7.03 (m, 34H), 6.75 (d, J ) 8,4 Hz,
2H), 6.69 (d, J ) 8.8 Hz, 2H), 4.98-4.63 (m, 16H), 4.46 (q, J )
12.1 Hz, 2H), 3.37 (q, J ) 9.2 Hz, 1H), 4.29 (t, J ) 2.2 Hz, 1H),
4.23 (ddd, J ) 2.6, 7.7, 9.9 Hz, 1H), 4.05 (t, J ) 9.5 Hz, 1H), 4.00
(t, J ) 9.9 Hz, 1H), 3.74 (s, 3H), 3.70 (s, 3H), 3.60 (dd, J ) 2.2,
9.9 Hz, 1H); 13C NMR (CDCl3, 100 MHz) δ 158.6, 151.9, 149.6,
138.4, 137.3, 135.8, 135.8, 135.4, 130.4, 130.1, 129.1, 128.9, 128.6,
128.3, 128.1, 128.1, 127.9, 127.8, 127.6, 127.5, 127.5, 127.3, 122.8,
113.3, 113.3, 96.5, 93.8, 90.8, 80.5, 78.6, 78.1, 78.0, 77.1, 77.1,
76.8, 76.6, 76.5, 76.3, 76.2, 75.0, 74.6, 74.2, 72.2, 69.4, 69.2, 69.1,
69.2; 31P NMR (CDCl3, 121 Hz) δ 1.1, -0.3, -0.4; IR (film, cm-1
)
3347, 3067, 3032, 2964, 2926, 2856, 1496, 1454, 1380, 1258; TLC
Rf 0.25 (60% ethyl acetate/hexanes); exact mass calcd for
[C63H65O16P3 + H]+ requires m/z 1171.3564, found 1171.3589
(ESI+). [R]D ) +0.8 (c 1.0, CH2Cl2).
55.1; 31P NMR (CDCl3, 121 Hz) δ -0.4, -0.8; IR (film, cm-1
)
3065, 3035, 2953, 2924, 2852, 2358, 2333, 1613, 1513, 1498, 1455,
1380, 1248, 1173, 1082, 1010, 881; TLC Rf 0.30 (50% ethyl acetate/
hexanes); exact mass calcd for [C65H68O15P2 + Na]+ requires m/z
1173.3931, found 1173.3937 (ESI+). [R]D ) +12.3 (c 1.0, CH2-
Cl2).
D-myo-Inositol-3,5,6-triphosphate Hexasodium Salt (D-I-3,5,-
6P3). To a stirred solution of 2-O-benzyl-1-O-benzyloxymethyl-
D-myo-inositol-3,5,6-tris(dibenzyl) phosphate (16, 12 mg, 0.010
mmol) in HPLC grade methanol (1.0 mL) was added 20%
palladium hydroxide on carbon (13 mg). The mixture was stirred
under hydrogen (atmospheric pressure) for 24 h and then filtered
through a syringe filter (Millex-HV 0.45 µm) to remove the catalyst.
Solvent was then removed under reduced pressure to afford D-I-
3,5,6P3 in free acid form. This material was redissolved in H2O
and stirred with 1 M NaOH (aq, 80 µL, 0.080 mmol). The solvent
was then removed via lyophilization and the resulting solid was
rinsed several times with HPLC grade methanol to afford D-I-3,5,-
6P3 as the hexasodium salt (5.0 mg, 90%). 1H NMR (D2O, pD 10,
300 MHz) δ 4.33 (br. s, 1H), 4.14 (q, J ) 7.8 Hz, 1H), 3.91-3.81
(m, 3H), 3.70 (dd, J ) 2.9, 9.8 Hz, 1H); 13C NMR (D2O, pD 10,
100 MHz) δ 78.0, 75.8, 75.4, 73.4, 72.6, 71.5; 31P NMR (D2O, pD
10, 121 Hz) δ 5.8, 5.7, 4.0; exact mass calcd for [C6H15O15P3 +
H]+ requires m/z 420.9702, found 420.9693 (ESI+); [R]D ) +9.8
(c 1.0, H2O, pH 10) [lit. (D-I-1,4,5P3) [R]D ) -20 (c 0.05, H2O,
pH 9),5c [R]D ) -3.19 (c 0.26, H2O, sodium salt),5b [R]D ) -24.1
(c 0.28, H2O, sodium salt)11]
2-O-Benzyl-1-O-benzyloxymethyl-D-myo-inositol-3,5-bis(diben-
zyl) Phosphate (15). To a stirred solution of 2-O-benzyl-1-O-
benzyloxymethyl-4,6-di-O-p-methoxybenzyl-D-myo-inositol-3,5-
bis(dibenzyl) phosphate (14, 130 mg, 0.113 mmol) in DCM (2.80
mL) was added distilled water (311 µL) followed by DDQ (77.0
mg, 0.339 mmol). The heterogeneous reaction was stirred at room
temperature for 12 h and then diluted with DCM and saturated
NaHCO3 solution was added. The reaction was then extracted with
DCM (4 × 50 mL) and the organic layers were combined, dried
(MgSO4), filtered and concentrated in vacuo. The resulting residue
was purified by column chromatography, eluting with 60% ethyl
acetate/hexanes to afford desired diol 15 (85.0 mg, 83%) as a white
solid. 1H NMR (CDCl3, 300 MHz) δ 7.43-7.18 (m, 30H), 5.18-
4.95 (m, 8H), 4.86-4.50 (m, 6H), 4.23-4.08 (m, 4H), 4.05 (s,
1H), 3.87 (d, J ) 1.8 Hz, 1H), 3.64 (d, J ) 1.6 Hz, 1H), 3.51-
3.41 (m, 1H); 13C NMR (CDCl3, 100 MHz) δ 138.2, 137.0, 135.5,
135.4, 128.4, 128.4, 128.3, 128.0, 127.8, 127.7, 127.3, 94.3, 82.6,
82.5, 78.5, 78.4, 77.7, 77.1, 75.0, 70.9, 70.8, 69.9, 69.8, 69.8, 69.6,
69.5; 31P NMR (CDCl3, 121 Hz) δ 1.1, 0.2; IR (film, cm-1) 3397,
3065, 3032, 2954, 2924, 2853, 1456, 1248, 1215, 1119; TLC Rf
0.27 (60% ethyl acetate/hexanes); exact mass calcd for [C49H52O13P2
Acknowledgment. We thank the National Institutes of
General Medical Sciences of the National Institutes of Health
for support (GM-068649).
+ Na]+ requires m/z 933.2781, found 933.2761 (ESI+). [R]D
+0.5 (c 5.0, CH2Cl2).
)
Supporting Information Available: Experimental procedures
and product characterization for additional intermediates discussed
in the manuscript. This material is available free of charge via the
Internet at http://pubs.acs.org.
2-O-Benzyl-1-O-benzyloxymethyl-D-myo-inositol-3,5,6-tris-
(dibenzyl) Phosphate (16). To a stirred solution of 2-O-benzyl-
1-O-benzyloxymethyl-D-myo-inositol-3,5-bis(dibenzyl) phosphate
(15, 50 mg, 0.055 mmol) in DCM (2.5 mL) was added dibenzyl
JO0610816
J. Org. Chem, Vol. 71, No. 18, 2006 6931