Regioselective phosphorylation of vicinal 3,4-hydroxy myo-inositol derivative promoted practical synthesis of D-PtdIns(4,5)P2 and D-Ins(1,4,5)P3

Article abstract of DOI:10.1016/S0040-4020(03)01212-2

The reactivity of 3 and 4-OH in 3,4-diol myo-inositol derivatives were observed through the phosphorylation, acylation and silylation. The results indicated that 3-OH is much more reactive than 4-OH, giving regiospecifically 3-mono-functionalized products

Full text of DOI:10.1016/S0040-4020(03)01212-2

Tetrahedron 59 (2003) 7703–7711
Regioselective phosphorylation of vicinal 3,4-hydroxy
myo-inositol derivative promoted practical synthesis
of D-PtdIns(4,5)P2 and D-Ins(1,4,5)P3
Fushe Han,a Minoru Hayashib and Yutaka Watanabeb
,
*
aVenture Business Laboratory, Ehime University, Matsuyama 790-8577, Japan
bDepartment of Applied Chemistry, Faculty of Engineering, Ehime University, Matsuyama 790-8577, Japan
Received 27 June 2003; revised 4 August 2003; accepted 4 August 2003
Abstract—The reactivity of 3 and 4-OH in 3,4-diol myo-inositol derivatives were observed through the phosphorylation, acylation and
silylation. The results indicated that 3-OH is much more reactive than 4-OH, giving regiospecifically 3-mono-functionalized products. This
investigation provided a concise methodology for the synthesis of natural D-form of PtdIns(4,5)P2 and D-Ins(1,4,5)P3 from L-1,2-O-
cyclohexylidene-3,4-O-(tetraisopropyl disiloxane-1,3-diyl)-myo-inositol.
q 2003 Elsevier Ltd. All rights reserved.
1. Introduction
These events have been stimulating vehemently chemists to
pursue a practical way to access these molecules. A number
of reports have been appeared for the synthesis of the
analogues of 1 from diverse starting materials, such as D-
glucose,6 myo-inositol,7 and L-(2)-quebrachitol.8 In spite of
these, many of the old synthetic procedures were not
entirely satisfactory, where long routes were required and
the overall yield was low. For 2, numerous routes hitherto
have been reported,9 among which two methods developed
by Potter10 and Ozaki11 are very rapid, involving only about
5 steps. However, a serious problem involved in these
methods is the formation of undesired regioisomers in
almost equal amount to the required ones during selective
protection–deprotection of hydroxyls, resulting in a low
yield for the target 2, although those undesired regioisomers
can provide starting materials for alternative targets. In
Potter’s approach (Scheme 1), for instance, selective
transformation of intermediate 3 yielded only about 31%
of the desired 4, accompanied by 40% of undesired
Since the discovery of the biological role of myo-inositol
1,4,5-trisphosphate [Ins(1,4,5)P3] as a second messenger
in intracellular signal transduction system in 1983,1 the
biological importance of inositol phosphates [Ins(P)ns] and
then phosphatidylinositol phosphates [PtdIns(P)ns] have
been well recognized in the past two decades. In the family
of PtdIns(P)ns, PtdIns(4,5)P2 (1) is the key member due to
its role as the precursor of at least three second-messenger
molecules,2 Ins(1,4,5)P3, PtdIns(3,4,5)P3 and diacylgly-
cerol. In addition, PtdIns(4,5)P2 itself may also be involved
in several other cellular processes such as exocytosis,
cytoskeletal regulation, and intracellular trafficking of
vesicles.3 Recent studies established PtdIns(4,5)P2 itself
is also a second-messenger.4 As for inositol phosphates,
Ins(1,4,5)P3 (2), that mediates the release of calcium ions
from intracellular stores,5 is the most important inositol
phosphate biologically.
Keywords: phosphorylation; synthesis; PtdIns(4,5)P2; Ins(1,4,5)P3.
*
Corresponding author. Tel.: þ81-89-927-9921; fax: þ81-89-927-9944;
e-mail: wyutaka@dpc.ehime-u.ac.jp
Scheme 1.
0040–4020/$ - see front matter q 2003 Elsevier Ltd. All rights reserved.
doi:10.1016/S0040-4020(03)01212-2

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F. Han et al. / Tetrahedron 59 (2003) 7703–7711
phosphites. Other electrophilic reactions such as benzoyl-
ation and silylation were also examined. In addition, to
expand the application of the practical methodology, we
studied the synthesis of D-Ins(1,4,5)P3 from L-11. The
synthetic procedure of dipalmitoyl PtdIns(4,5)P2 1 is also
detailed.
Scheme 2.
2. Results and discussion
regioisomer 5. Similarly, 41% of desired 9 and 48% of
undesired 10 were formed, respectively, in Ozaki’s method
(Scheme 2), risen from the poor selectivity of acetylation
of tetrol 6 to give inseparable mixture of diol 7 and 8.
Therefore, developing more practical methods for the
syntheses of PtdIns(4,5) and Ins(1,4,5)P3 are still
significant.
2.1. Phosphorylation of various vicinal 3,4-diols (12)
Although the phosphorylation of hydroxyls in 1,2-diol
system of suitably protected myo-inositols have been well
documented to give regiospecifically 1-O-phosphates with
good yields,7a,17 the selectivity of hydroxyls in 3,4 (1,6)-diol
inositol derivatives is unpredictable based on these results
because both 3 and 4-OH are equatorial hydroxyls to
contrast with equatorial/axial 1,2-dihydroxyls, where 2-OH
shows poor reactivity due to the sterically hindered axial
orientation.18 On the other hand, some reports19 indicated
that 3-OH on 1,2-ketal 6 and 1,2:5,6-diketal is more reactive
than other OHs in their electrophilic reactions such as
acylation, silylation and alkylation. Although some reports
showed completely different results where the selectivity is
poor,20,21 even in some cases 4- and/or 5-OH are more
reactive.11,22 To facilitate the synthesis of dipalmityl
PtdIns(4,5)P2 (1) and Ins(1,4,5)P3 (2), as well as to utilize
the formerly ‘wasted’ L-11, we therefore studied the
phosphorylating reaction of 3,4-diol 12 (racemic materials
were used here), such reaction has rarely been investigated
previously. Preliminary study on the phosphorylation of 12a
with 1,2-di-O-palmitoylglycerol phosphite 13a in the
presence of pyridinium tribromide regiospecifically
occurred at the 3-OH position to give 3-monophosphate
14a in excellent yield (Table 1, entry 1). To define the scope
On the other hand, we have demonstrated that 1,2-O-
cyclohexylidene-3,4-O-(tetraisopropyldisiloxane-1,3-diyl)-
myo-inositol (11) may be a pivotal intermediate for the
synthesis of diverse inositol phosphates such as Ins(4)P,12a
Ins(5)P,12a Ins(1,3,4)P3,12a and phosphatidylinositol phos-
phates such as PI(5)P2,12b PI(3,4)P2,13 PI(3,4,5)P3,14 and
PIM2.15 In these cases, D-11 was derived into the target
molecules with the natural configuration. The opposite L-11
has been the ‘waste’ one until now, whereas both
enantiomers can be readily derived from the selective
disiloxanylation with 1,3-dichlorotetraisopropyl-disil-
oxane14b of the corresponding D and L-1,2-O-cyclohexyl-
idene-myo-inositol obtained by the enzyme-aided optical
resolution11b with excellent yields (higher than 42%
from myo-inositol, respectively). The utilization of L-11
(Scheme 3) and the practical synthesis of 1 and 2 are of
particular interest to us. These challenges have been
accomplished in the synthesis of dipalmitoyl PtdIns(4,5)P2
(1) from L-11,16 where regioselective phosphorylation of
1,2-O-cyclohexylidene-5,6-dis-O-(o-xylylene phosphoryl)-
myo-inositol L-12a at 3-position, as a key reaction, provided
a very rapid route to get 1. We are interested in the
extremely selective phosphorylation, and investigated
furthermore the selectivity in the phosphorylation of
various 3,4-diol derivatives of myo-inositol with different
Table 1. Phosphorylation of racemic diol 12 with various phosphites
Entry
Diol
Phosphite
(13)
P in compound 14
yield
(%)
a
DAG-P(OBn)2 13a
1
2
12a
12a
DAG-P(O)(OBn) 14a
86
63
3
4
12a
12a
12a
12b
12c
12d
12e
DAG-P(OBn)2 13c
DAG-P(OMe)2 13d
P(OBn)3 13e
13e
13e
13e
13e
DAG-P(O)(OBn) 14c
DAG-P(O)(OBn) 14d
P(O)(OBn)2 14e
P(O)(OBn)2 14f
P(O)(OBn)2 14g
P(O)(OBn)2 14h
P(O)(OBn)2 14i
88
89
91
89
91
86
86
5
6b
7
8
9
General reaction conditions: phosphite (3 equiv.), 2,6-lutidine (5 equiv.),
pyridinium tribromide (PTB) (4 equiv.), 2428C, 10 min then 08C, 1.5–
2.0 h, solvent CH2Cl2 unless otherwise noted.
a
b
Scheme 3.
Solvent: CH2Cl2/Py (v/v 3:1).

F. Han et al. / Tetrahedron 59 (2003) 7703–7711
7705
of this reaction, we investigated the phosphorylation of 12a
with various phosphites 13b-e using the phosphite-pyri-
dinium tribromide approach. The reaction proceeded
exclusively at the OH-3 position in all cases as listed in
Table 1 (entries 2–5), without the formation of the another
regioisomer and/or bisphosphorylated product as analyzed
by TLC and NMR. The phosphorylation position was
determined by transforming the phosphates 14 (except for
14e) into their 4-O-chloroacetyl derivatives because signals
corresponding to InsH-4 overlapped with those of a and
1
g-methylene protons in the glycerol moiety. The H NMR
1
spectra, combined with the H–1HCOSY, clearly showed
that in each case the chemical shift of InsH-4 shifted
downfield about 1.4 ppm. The results, summarized above,
suggest that other vicinal 3,4-diol compounds might also
show a good selectivity. To test this prediction, we have
examined the phosphorylation reaction of various 3,4-diols
bearing different substituents at 5- and 6-positions (12b-e)
with tribenzyl phosphite 13e (entries 6–9). As expected, in
all cases, only 3-mono-phosphorylated products, 14f-i were
isolated in high yields, indicating the selectivity is less
affected by substituents at 5- and 6-positions.
Figure 1. (a) Conformation of L-12a deduced from the coupling constants
and MM-2 calculation as a represntative example of 3,4-diol monoketal
derivatives 12a-e; (b) chair-form conformation.
form the coupling constants and MM-2 calculation
(Fig. 1(a)) suggests that it rather takes a boat-like one,
furthermore, the 3-OH flipped to take an axial-like
orientation. Such a conformation makes the 3-OH less
crowded, but 4-OH sterically hindered to compare with a
chair-form one (Fig. 1(b)), resulting in the higher reactivity
of 3-OH than that of 4-OH. As a support experiment to
examine our hypothesis, we conducted the phosphorylation
of 1,2:5,6-diketal 12g, which has been well defined to
take a boat-form conformation because of the locked ring
system.20 Similarly, only 3-mono-phosphorylated product
14j was obtained in high yield in the presence of 3 equiv.
of phosphite 13e. Whilst the selective functionalization of
1,2:4,5-diketal such as 12f and dicyclohexylidene derivative
was not always good,20,23 di-functionalizd and/or 6-mono-
functionalized by-product were isolated, in addition to the
3-mono-functionalized major product. Since 1,2:4,5-diketal
can also form intramolecular hydrogen bond,20 the selec-
tivity differences between the boat-like 3,4-diol and chair-
form 1,2:4,5-diketal may be caused by their different
conformations. Thus, intramolecular hydrogen bonding as
well as the favourable conformation may be the factors to
influence the reactivity of 3- and 4-OH. Whilst the main
reason is yet not clear.
According to the literature,20 such a good selectivity is
probably due to the intramolecular hydrogen bonding
between the 3-OH and oxygen at C-2, which results in the
enhancement of the acidity of 3-OH. In addition, we thought
that another plausible explanation for the high reactivity of
3-OH in vicinal 3,4-diol derivatives is due to their
conformations. By comparison the vicinal coupling con-
stants of inositol methine protons of 3,4-diol 12a-e with
those of the already reported chair-form 3,4-diketal 12f20
(Scheme 4 and Table 2), conformations of the all investi-
gated 3,4-diol monoketal 12a-e were apparently deviated
from a normal chair-form. Conformation of 12a-e deduced
2.2. Benzoylation and silylation of 3,4-diol (Scheme 5)
The remarkably different reactivity between 3- and
4-hydroxyls in vicinal 3,4-diol derivatives in the phos-
phorylation is of interest, and therefore intrigued us to study
Scheme 4.
Table 2. Coupling constants of some representative examples of diol 12
observed in CDCl3
Compounds
Coupling constants (J, Hz)
H1–H2 H2–H3 H3–H4 H4–H5 H5–H6 H6–H1
12a
12e
5.6
5.6
4.8
6.2
3.7
4.0
4.8
4.2
7.8
8.8
9.4
5.3
7.4
8.8
9.4
9.1
8.8
9.5
10.5
10.2
6.4
7.2
6.4
8.1
12fa
12ga
a
For the coupling constants, see also Ref. 20
Scheme 5.

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F. Han et al. / Tetrahedron 59 (2003) 7703–7711
Table 3. Benzoylation and silylation of vicinal 3,4-diol 12a
Entry
Reaction conditions
Yield (%) of the productsa
S. M.b (%)
NDd
1
2
3
4
C6H5COCl (1.2 equiv.), Py (3.0 equiv.), DMAPc (cat.), 08C, 5.3 h
C6H5COCl (2.0 equiv.), Py (3.0 equiv.), DMAP (cat.), 08C, 4 h
t-BuMe2SiCl (1.1 equiv.), imidzole (3.3 equiv.), DMF, rt, 16 h
t-BuMe2SiCl (1.1 equiv.), TBMPf (3.0 equiv.), DMAPf (cat.), CH2Cl2, rt,
16 h
Et3SiCl (1.05 equiv.), EDAg (3.0 equiv.), DMAP (cat.), CH2Cl2, rt, 14 h
Et3SiCl (1.05 equiv.), EDA (3.0 equiv.), DMAP (cat.), CH2Cl2, 08C, 4.5 h
15a (63)
15a (87)
15b (1)
15b (2)
16a/16be (total yield 47%)
16a/16b (total yield 51%)
ND
ND
5
6
17a (18.0)
17a (21)
17b (24)
17b (21)
28
33
a
Isolated by column chromatography unless noted otherwise.
Starting material recovered.
DMAP: N,N-dimethylamino pyridine.
Not determined.
Inseparable regioisomer, the ratio of the regioisomer 16a/16b was about 2:1 as determined by 1H NMR analysis.
TBMP: 2,6-t-butyl-4-methyl pyridine.
EDA: ethyldiisopropylamine.
b
c
d
e
f
g
other electrophilic substitution reactions. Herein diol 12a
was chosen as a representative example of mono-ketal 12a-e
derivatives to study the functionalization such as acylation
and silylation (Scheme 5). As outlined in Table 3, acylation
of 12a with benzoyl chloride carried out in different
conditions has good selectivity (entries 1 and 2). Reaction
carried out at 08C occurred regiospecifically at 3-OH to give
15a even if 2 equiv. of the chloride was used (entry 2).
Silylation of 12a with t-BuMe2SiCl under different
conditions gave inseparable regioisomers 16a/16b (entries
3 and 4). The ratio was about 2:1 as determined by the
1H NMR spectra. Silylation with Et3SiCl gave almost 1:1
separable regioisomers of 17a/17b (entries 5 and 6). No
disilylated product was detected in each case. The
selectivity of silylation is poor, whilst that of acylation is
quite good. The significant selectivity differences between
the acylation and silyaltion led us to look into the reasons.
Control experiment showed that migration of silyl groups in
both pure 17a or 17b occurred to form about 1:1 mixture
(monitored by TLC and NMR) of 17a and 17b in the
presence of EDA and DMAP in CH2Cl2. Cat. amount of
DMAP played the key role in promoting the migration. Such
results suggested that the poor selectivity for the silylation
of 12a was indeed caused by the migration of silyl group
in the real reaction system. This result may elucidate the
previous experimental results that the selectivity for the
silylation of 3- and 4-OH in diketal 12g was also low.20
2.3. Synthesis of dipalmitoyl D-PtdIns(4,5)P2 (1)
and D-Ins(1,4,5)P3 (2) (Scheme 6)
The outstanding regioselectivity of 3-OH in 3,4-diols 12a-e
provides a convenient route for the synthesis of
PtdIns(4,5)P216 and Ins(1,4,5)P3. Thus, L-11 was trans-
Scheme 6. Reagents and conditions: (a) o-xylylene N,N-diethyl phosphoramidite (XEPA), 1H-tetrazole, CH2Cl2, then mCPBA, 95%; (b) TBAF·3H2O, HOAc,
THF, 2208C, 81%; (c) PyHBr3 (PTB), 2,6-lutidine, CH2Cl2, 242 to 08C, 88%; (d) Py(HF)n, CH2Cl2, 08C to rt, 1.5 h, 92%; (e) 10%-Pd/C-H2 (25 wt%),
EtOH/CHCl3 (1:1), rt, overnight, quant.; (f) 10%–Pd/C–H2, EtOAc, rt, 2.5 days, quant.; (g) (BnO)3P, 2,6-lutidine, PTB, 242 to 08C, 91%; (h) 10%–Pd/C–H2
(25 wt%), EtOH; then H2O/HOAc (2:1), 2 days, quant.; (i) 10%–Pd/C–H2 (25 wt%), tributylamine (2 equiv.), MeOH/CHCl3 (1:1), 3 days, quant.

F. Han et al. / Tetrahedron 59 (2003) 7703–7711
7707
formed into the diol L-12a through the phosphorylation with
o-xylylene N,N-diethyl-phosphoramidite (XEPA)24 to give
bisphosphate 18, followed by the desilylation. Phosphoryl-
ation of L-12a with 1,2-di-O-palmitoyl-sn-glycerol phos-
phite 13a afforded D-14a as described above. With
compound D-14a in hand, the final product, D-dipalmitoyl
PtdIns(4,5)P2 1 might be obtained by removal of the
cyclohexylidene and benzylic groups. The cleavage of the
cyclohexylidene ketal could be achieved smoothly by using
pyridinium poly(hydrogen fluoride), which was used to
decompose the isopropylidene group without the migration
of the adjacent phosphate function.25 The triol 19 thus
obtained was hydrogenated to afford dipalmitoyl 1. We
eventually found that such a two-step strategy might be
combined into a one-step procedure when the hydro-
genolysis of D-14a was performed in a commercial grade
EtOAc, resulting in the simultaneous removal of the
cyclohexylidene ketal to give dipalmitoyl 1 in quantitative
yield as its triethylammonium salt. No other undesired
products were detected by TLC and spectral analyses.
A comparison of EtOAc to MeOH suggested that an aprotic
solvent was much more efficient than a protic one in
promoting the spontaneous removal of the cyclohexylidene
ketal, probably due to the protic solvent solvating the proton
of the phosphoric acid, as a result, the catalytic ability of the
phosphoric acid was decreased.
not the shortest one, it refrains from the formation of the
undesired regioisomers as appeared in the old methods, and
thus makes this method advantageous.
3. Experimental
3.1. General methods
All solvents were purified according to the standard
procedures. Reagents were reagent grade, and purified
where further purification being required. Pyridinium
tribromide (PTB) was purified by recrystallization from
AcOH. 2,6-Lutidine, 1,3-dichlorotetraisopropyldisiloxane,
benzoyl chloride and Et3SiCl were purified by distillation
under reduced pressure. NMR spectra were recorded on
Bruker Avance 400 unless otherwise noted. Optical rotation
was measured using JASCO P-1010 with 1 cm cell.
Elemental analyses were performed on Perkin–Elmer
240C. Melting points were uncorrected and tested in open
capillaries by using Yamato Melting Point Apparatus Model
MP-21. Silica gel (Fuji Silysia Chemical Ltd., 200–400
mesh) was used for flash chromatography.
3.2. A general procedure for the phosphorylation of a
diol 12 with a phosphite
In a similar manner, Ins(1,4,5)P3, 2 was synthesized as
follows. Phosphorylation of L-12a using tribenzyl phosphite
13e afforded D-14e in 91% yield without formation of the
other isomers. Spontaneous removal of cyclohexylidene
ketal and benzylic groups in D-14e was first tried in EtOAc,
however, it was not effective because the intermediates
partially or completely hydrogenated would precipitate out
from the solvent. Bruzik’s26 protocol employing MeOH as
solvent was also not satisfactory. Several solvent system
such as MeOH/EtOAc/triethylamine, MeOH/CHCl3/
triethylamine, MeOH/EtOAc/tributylamine and MeOH/
CHCl3/tributylamine were scanned. We finally found that
when the hydrogenolysis was performed in a mixture of
CHCl3 and MeOH (v/v¼1:1) in the presence of 2 equiv. of
tributylamine, the cyclohexylidene ketal could be cleaved
simultaneously to afford 2 completely. In this case, the
amine and MeOH were necessary to dissolve the inter-
mediates partially or completely hydrogenated in the
medium. The ammonium salt of 2 obtained from our one-
step procedure was transformed into its sodium salt by
passing through the sodium form of ion-exchange resin.
Alternatively, removal of benzylic groups followed by the
cleavage of cyclohexylidene ketal using Vacca’s protocol27
was also applicable once the cleavage of cyclohexylidene
ketal was performed at a 1:2 (v/v) H2O/HOAc system,
although the essential condition was proved to be difficult to
cleave the ketal completely.
To a CH2Cl2 solution of a diol 12 and phosphite 13
(3 equiv.) were added dropwise 2,6-lutidine (5 equiv.) and
pyridinium tribromide (PTB, 4 equiv.) at 2428C. The
solution was stirred vigorously at the same temperature
for 10 min, and then at 08C (an ice/H2O including a small
amount of NaCl bath) for about 2 h. After being diluted with
EtOAc, the solution was washed with aq. KHSO4, aq.
NaHCO3 and brine. The organic layer was dried, con-
centrated, and chromatographed (hexane/EtOAc¼1:2) to
afford 14.
3.2.1. Compound 14a. The title compound (86%) as
colorless gum. Rf¼0.65 (hexane/EtOAc¼1:3); 1H NMR
(400 MHz) d: 7.31–7.39 (m, 13H), 5.54–5.65 (complex,
2H, ArCH2O), 5.30 (t, 1H, J¼13.6 Hz, ArCH2O), 5.04–
5.23 (complex, 8H in ArCH2O, glyceryl 2-H), 4.85 (br 2£q,
1H, J¼9.0 Hz, InsH-6), 4.64–4.71 (complex, 1.5H, InsH-3
and H-2), 4.60 (br 2£t, 0.5H, J¼4.0 Hz, InsH-2), 4.39 (br
2£dd, 1H, J¼4.4, 9.0 Hz, InsH-5), 4.11–4.35 (complex,
6H, InsH-1, H-4, glyceryl H-1, H-3), 2.28 (m, 4H, Pal H-2),
1.82 (m, 2H, cyclohexylidene H), 1.53–1.67 (br, 12H, Pal
3-H, cyclohexylidene H), 1.25 (br, 48H, Pal H4-15), 0.88 (t,
6H, J¼6.2 Hz Pal CH3); 13C NMR (100.6 MHz) d: 173.73
(2C), 173.59 (2C), 173.46, 173.33, 173.22, 173.17 (Pal CO),
128.30–135.89 (complex, aromatic C), 112.38 and 112.37
(spiral C), 79.57–80.13 (complex, InsC-5 C-6), 75.90–
76.70 (m, InsC-3), 74.57 and 74.49 (m each, InsC-2),
68.63–71.80 (complex, InsC-4, glyceryl 2-C and ArCH2O),
65.90–66.34 (m, glyceryl 3-C), 61.80–62.30 (m, glyceryl
1-C), 37.90 and 37.86 (cyclohexylidene C), 35.44, 34.40–
34.55 (complex, Pal C-2), 32.31 (complex, Pal C-14),
29.49–30.10 (complex, Pal C4-13), 25.22 (complex, Pal
C-3, cyclohexylidene C), 24.06 and 24.21 (s each,
cyclohexylidene C), 23.08 (s, Pal C-15), 14.51 (s, Pal
C-16), signals for InsC-1 was overlapped with those of
CDCl3; 31P NMR (162 MHz) d: 20.21, 20.23, 20.27 (s
In summary, the finding as of the remarkable selective
functionalization of 3-OH in vicinal 3,4-diol system, as well
as the one-step deprotection of ketal and benzylic groups in
the final stage, provided a practical methodology for the
synthesis of dipalmitoyl PtdIns(4,5)P2 (1) and Ins(1,4,5)P3
(2) from L-11. It is the shortest and more practical method
for the synthesis of 1 compared with the reported methods.
For Ins(1,4,5)P3 (2), although this method presented here is

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F. Han et al. / Tetrahedron 59 (2003) 7703–7711
each, 1P), 20.31, 20.44, 20.47 (s each, 1P), 20.50, 20.56,
20.61 (s each, 1P); Anal. Calcd. for C70H107O19P3: C,
62.48; H, 8.02; found: C, 62.10; H, 8.02.
J¼0.9 Hz, InsC-2), 70.79 (d, J¼5.1 Hz, InsC-4), 69.85 (d,
J¼5.7 Hz), 69.52 (d, J¼5.8 Hz), 69.31 and 69.12 (2£d, J¼
7.3 Hz), 68.90 (d, J¼7.0 Hz), 68.68 (d, J¼7.0 Hz)
(ArCH2O), 37.50 (s), 34.98 (s), 29.70 (s), 24.85 (s),
23.82 (s), 23.69 (s) (cyclohexylidene); 31P NMR
(162 MHz) d: 20.23 (s), 20.27 (s), 20.46 (s). Anal.
Calcd. for C42H47O15P3: C, 57.02; H, 5.35; found: C,
56.84; H, 5.47.
3.2.2. Compound 14b. The title compound (63%) as
colorless gum. Rf¼0.66 (hexane/EtOAc¼1:3); 1H NMR
(400 MHz) d: 7.29–7.39 (m, 13H), 5.53–5.64 (complex,
2H, ArCH2O), 5.29 (t, 1H, J¼13.2 Hz, ArCH2O), 5.08–
5.28 (complex, 8H, ArCH2O, glyceryl 2-H), 4.85 (br 2£q,
1H, J¼9.0 Hz, InsH-6), 4.64–4.73 (complex, 1.5H, InsH-3,
H-2), 4.54–4.63 (complex, 2.5H, InsH-2, CH2Br), 4.40
(dd£2, 1H, J¼4.0, 9.0 Hz, InsH-5), 4.01–4.36 (complex,
6H, InsH-1, H-4, glyceryl 1-H, 3-H), 2.28 (complex, 4H, Pal
H-2), 1.82 (br, 2H, cyclohexylidene H), 1.38–1.64 (br, 12H,
Pal H-3, cycohexylidene H), 1.25 (br, 48H, Pal H4-15), 0.89
(t, 6H, J¼6.8 Hz, Pal CH3); 31P NMR (162 MHz) d: 20.20,
20.22, 20.27 (s each, 1P), 20.32, 20.45, 20.48 (s each,
1P), 20.52, 20.59, 20.63 (s each, 1P).
3.2.6. Compound 14f. The title compound (89%) as white
1
solid. Rf¼0.38 (hexane/EtOAc¼1:3); H NMR (400 MHz)
d: 7.34 (m, 14H, aromatic H), 5.28–5.37 (complex, 3H,
InsH-6 and ArCH2O), 5.06–5.19 (complex, 6H, ArCH2O),
4.56 (dt, 1H, J¼4.0, 9.2 Hz, InsH-3), 4.47 (dd, 1H, J¼4.0,
4.8 Hz, InsH-2), 4.38 and 4.36 (t£2, 1H, J¼9.2 Hz, InsH-5),
4.26 (t, 1H, J¼9.2 Hz, InsH-4), 4.12 (dd, 1H, J¼4.8, 6.8 Hz,
InsH-1), 2.67–2.76 (complex, 4H, CH2CH2), 2.15 (s, 3H,
CH3), 1.81 (complex, 2H, cyclohexylidene), 1.63 (br, 2H,
cyclohexylidene), 1.50 (br, 4H, cyclohexylidene), 1.26 (br,
2H, cyclohexylidene); 31P NMR (162 MHz) d: 0.29 (1P),
0.01 (1P).
3.2.3. Compound 14c. The title compound (88%) as
colorless gum. Rf¼0.60 (hexane/EtOAc¼1:3); 1H NMR
(400 MHz) d: 7.30–7.39 (m, 13H), 5.55–5.65 (complex,
2H, ArCH2O), 5.28 (t, 1H, J¼13.2 Hz, ArCH2O), 5.07–
5.27 (complex, 8H, ArCH2O, glyceryl 2-H), 4.85 (br 2£q,
1H, J¼9.0 Hz, InsH-6), 4.64–4.73 (complex, 1.5H, InsH-3,
H-2), 4.54 and 4.59 (2£t, 0.5H, J¼4.0 Hz, InsH-2), 4.40
(2£dd, 1H, J¼4.0, 9.0 Hz, InsH-5), 4.08–4.36 (complex,
6H, InsH-1 and H-4, glyceryl H-1, H-3), 2.28 (complex, 4H,
Oct H-2), 1.82 (br, 2H, cyclohexylidene H), 1.38–1.64 (br,
12H, Oct H-3, cycohexylidene H), 1.25 (br, 16H, Oct H3-6),
0.87 (t, 6H, J¼6.8 Hz, Oct CH3); 31P NMR (162 MHz) d:
20.25, 20.27, 20.32 (s each, 1P), 20.33, 20.46 (0.5P)
(s each, 1P), 20.58, 20.64 (s each, 1P).
3.2.7. Compound 14g. The title compound (91%) as white
solid. Rf¼0.35 (EtOAc/hexane¼4:1); mp 155.5–157.08C
(sample was obtained by chromatography); 1H NMR
(400 MHz) d: 7.35 (m, 20H, 4£C6H5), 5.26 (dd, 1H, J¼
7.4, 8.8 Hz, InsH-6), 4.50–5.19 (complex, 8H, 4£C6H5CH2),
4.44–4.50 (complex, 2H, InsH-3, H-2), 4.13–4.23 (com-
plex, 2H, InsH-4, H-5), 4.04 (dd, 1H, J¼4.8, 7.4 Hz,
InsH-1), 2.44–2.57 (complex, 4H, CH2CH2), 2.06 (s, 3H,
CH3), 1.60 (br, 2H, cyclohexylidene), 1.48 (br, 6H, cyclo-
hexylidene), 1.33 (br, 2H, cyclohexylidene); 13C NMR
(100.6 MHz) d: 206.52 (s), 172.17 (s), 136.16 (s), 136.08
(s), 136.01 (s), 135.94 (s), 128.26–129.50 (m), 112.01 (s),
79.04 (br), 76.50 (br), 75.93 (m), 74.56 (s), 73.91
(t, J¼3.3 Hz), 70.28 (s), 70.23 (s, 2C), 70.17 (s), 69.97 (d,
J¼5.5 Hz), 38.04 (s), 37.69 (s), 35.39 (s), 30.10 (s), 28.29
(s), 25.34 (br), 24.15 (s), 23.98 (s); 31P NMR (162 MHz) d:
0.45 (1P), 20.05 (1P); calcd for C45H52O14P2·0.5H2O: C,
60.87; H, 6.02; found: C, 61.02; H, 6.24.
3.2.4. Compound 14d. The title compound (89%) as
colorless gum: Rf¼0.59 (hexane/EtOAc¼1:3); 1H NMR
(400 MHz) d: 7.28–7.37 (m, 8H), 5.57–5.63 (complex, 2H,
ArCH2O), 5.30 (t, 1H, J¼13.0 Hz, ArCH2O), 5.03–5.25
(complex, 6H, ArCH2O glyceryl 2-H), 4.85 (br 2£q, 1H,
J¼8.8 Hz, InsH-6), 4.52–4.68 (complex, 2H, InsH-3, H-2),
4.42 (2£dd, 1H, J¼4.0, 8.8 Hz, InsH-5), 4.12–4.30
(complex, 6H, InsH-1 and H-4, glyceryl 1-H, 3-H), 3.79–
3.87 (complex, 3H, OMe), 2.32 (complex, Oct H-2), 1.82
(br, 2H, cycohexylidene H), 1.40–1.65 (br, 12H, Oct H-3,
cycohexylidene, H), 1.28 (br, 16H, Oct H4-6), 0.86 (t, 6H,
J¼7.2 Hz, Oct CH3); 31P NMR (162 MHz) d: 0.59, 0.54,
0.52, 0.42 (s each, 1P), 20.23, 20.28, 20.31 (s each, 1P),
20.57, 20.58, 20.63 (s each, 1P).
3.2.8. Compound 14h. The title compound (86% yield) as
colorless gum. Rf¼0.43 (hexane/AcOEt¼1:2); 1H NMR
(400 MHz) d: 7.28–7.47 (m, 28H, aromatic), 6.08 (s, 1H,
methenyl), 6.05 (s, 1H, methenyl), 5.38 (dd, 2H, J¼7.6,
9.2 Hz, InsH-6), 5.27–5.34 (complex, 2H, ArCH2O), 4.71–
5.18 (complex, 14H, ArCH2O), 4.50–4.57 (complex, 3H,
InsH-3 and InsH-2), 4.39 (dd, 1H, J¼4.0, 4.8 Hz, InsH-2),
4.28–4.36 (complex, 4H, InsH-5 and InsH-4), 4.22 (dd,
1H, J¼4.8, 7.6 Hz, InsH-1), 3.79 (dd, 1H, J¼4.8, 7.6 Hz,
InsH-1), 2.18 (s, 3H, CH3), 2.12 (s, 3H, CH3), 1.75 (br, 2H,
cyclohexylidene), 1.48–1.61 (br, 6H, cyclohexylidene),
1.31 (br, 2H, cyclohexylidene); 31P NMR (162 MHz) d:
0.36 (1P), 20.19 (1P), 20.27 (2P).
3.2.5. Compound 14e. The title compound (91%) as white
1
solid. Rf¼0.41 (hexane/EtOAc¼1:3); H NMR (400 MHz)
d: 7.31–7.39 (m, 18H), 5.53–5.62 (complex, 2H, ArCH2O),
5.30 (t, 1H, J¼13.6 Hz, ArCH2O), 5.01–5.22 (complex, 9H,
ArCH2O), 4.85 (dt, 1H, J¼7.4, 9.2 Hz, InsH-6), 4.60 (dd,
1H, J¼4.8, 9.0 Hz, InsH-3), 4.55 (t, 1H, J¼4.8 Hz, InsH-2),
4.39 (dt, 1H, J¼7.4, 9.2 Hz, InsH-5), 4.29 and 4.30 (2£t,
1H, J¼7.4, 9.0 Hz, InsH-4), 4.22 (dd, 1H, J¼4.8, 7.4 Hz,
InsH-1), 1.82 (br, 2H, cyclohexylidene H), 1.51–1.64 (br,
6H, cyclohexylidene H), 1.25 (br, 2H, cyclohexylidene H);
13C NMR (100.6 MHz) d: 135.08–135.73 (m, aromatic C),
127.92–129.21 (m, aromatic C), 112.43 (s, spiral C), 79.65
(t, J¼5.8 Hz, InsC-6), 79.23 (t, J¼5.1 Hz, InsC-5), 76.30 (d,
J¼2.2 Hz, InsC-1), 75.81 (d, J¼5.8 Hz, InsC-3), 74.09 (d,
3.2.9. Compound 14i. The title compound (86%) as
1
white solid. H NMR (400 MHz) d: 7.29–7.37 (m, 10H,
aromatic), 5.22 (dd, 1H, J¼7.2, 9.6 Hz, InsH-6), 5.01–5.15
(complex, 4H, ArCH2O), 4.83 (dd, 1H, J¼9.2, 9.6 Hz,
InsH-5), 4.51 (dt, 1H, J¼4.0, 9.2 Hz, InsH-3), 4.44 (dd, 1H,
J¼4.0, 5.0 Hz, InsH-2), 4.14 (t, 1H, J¼9.2 Hz, InsH-4), 4.10
(dd, 1H, J¼5.0, 7.2 Hz, InsH-1), 2.07 (s, 3H, CH3), 2.06 (s,
3H, CH3), 1.79 (m, 2H, cyclohexylidene), 1.63 (br, 2H,

F. Han et al. / Tetrahedron 59 (2003) 7703–7711
7709
cyclohexylidene), 1.61 (br, 4H, cyclohexylidene), 1.32
(br, 2H, cyclohexylidene); 31P NMR (162 MHz) d: 20.33
(1P).
Entry 2. The reaction was carried out following the same
procedure as entry 1 at 08C for 4 h in the presence of
2.0 equiv. of benzoyl chloride. 86.5% yield for 15a.
3.2.10. Phosphorylation of 1,2:5,6-di-O-isopropylidene
myo-inositol (12g). To a pyridine (4 mL) and CH2Cl2
(6 mL) solution of 12g (100 mg, 0.292 mmol) was added
2,6-lutidine (171.6 mL, 1.462 mmol), tribenzyl phosphite
13e (308.8 mg, 0.877 mmol) and pyridinium tribromide
(PTB, 374.3 mg, 1.170 mmol) at 2428C. The mixture was
stirred at the same temperature for 10 min, then 08C for 2 h.
After being diluted with EtOAc, the solution was washed
with KHSO4 aq., NaHCO3 aq., and brine, the organic layer
was dried, concentrated and chromatographed (EtOAc/
hexane¼1:1) to afford 14j (169 mg, 85%; gum solidifies
on standing). Rf¼0.33 (EtOAc/hexane¼2:1); 1H NMR
(400 MHz) d: 7.36 (m, 10H, 2£C6H5), 5.03–5.14 (complex,
4H, 2£C6H5CH2), 4.48 (dd, 1H, J¼4.4, 5.6 Hz, InsH-2),
4.21 (ddd, 1H, J¼4.4, 6.4, 8.8 Hz, InsH-3), 4.24 (dd,
1H, J¼5.6, 8.4 Hz, InsH-1), 4.16 (dd, 1H, J¼6.4, 10.0 Hz,
InsH-4), 3.79 (dd, 1H, J¼8.4, 10.0 Hz, InsH-6), 3.33 (t,
1H, J¼10.0 Hz, InsH-5), 1.47 (s, 3H, CH3), 1.43 (s, 6H,
2£CH3), 1.30 (s, 3H, CH3); 13C NMR (100.6 MHz) d:
135.58, 135.54, 135.50, 135.47, 128.72, 128.68, 128.65,
128.63, 128.54, 128.44, 128.06, 128.03 (aromatic C),
112.58, 110.78 (spiral C), 79.89 (d, J¼6.3 Hz), 78.32 (s),
77.32 (s), 76.07 (s), 75.31 (d, J¼3.8 Hz), 71.80 (d, J¼
4.1 Hz) (InsC-1-C-6), 69.86 (d, J¼5.6 Hz), 69.72 (d, J¼
5.6 Hz) (CH2), 27.88 (s, CH3), 26.88 (s, 2C, CH3), 25.64
(s, CH3); 31P NMR (162 MHz) d: 0.39. calcd for
C26H33O9P·0.5H2O: C, 58.97; H, 6.47; found: C, 58.86, H,
6.30.
3.2.12. Silylation of 1,2-O-cyclohexylidene-5,6-bis-O-
(o-xylylene phosphoryl)-myo-inositol 12a. Entry 3. To a
DMF solution (1.5 mL) of 12a (80 mg, 0.128 mmol) was
added imidazole (28.7 mg, 0.422 mmol) and t-BuMe2SiCl
(21.2 mg, 0.141 mmol) at 08C. The mixture was then
stirred at rt for 16 h. After being diluted with EtOAc, the
solution was washed with KHSO4 aq., NaHCO3 aq., and
brine, the organic layer was dried, concentrated and
chromatographed (EtOAc/hexane¼1:1) to give the inse-
parable regioisomers of 16a and 16b (44.6 mg, 47%) as
white solid. The ratio of the products was determined by
1
their H NMR spectra.
Entry 4. To a CH2Cl2 solution (2.4 mL) of 12a (48.6 mg,
0.078 mmol) was added t-BuMe2SiCl (12.8 mg,
0.086 mmol) and 2,6-t-butyl-4-methylpyridine (TBMP
47.8 mg, 0.233 mmol) and at 08C. The mixture was then
stirred at rt for 16 h. Workup with the same procedure as
entry 4 gave the mixture of 16a and 16b (39.3 mg, 51%) as
white solid.
Entry 5. To a CH2Cl2 solution (3 mL) of 12a (102 mg,
0.163 mmol) was added diisopropylethylamine (EDA)
(63.2 mg, 0.490 mmol), DMAP (cat.) and Et3SiCl
(24.5 mg, 0.163 mmol) at 08C. The mixture was then stirred
at rt for 16 h. After being diluted with EtOAc, the solution
was washed with KHSO4 aq., NaHCO3 aq., and brine, the
organic layer was dried, concentrated and chromatographed
(EtOAc/hexane¼1:1) to give 17a (21.3 mg, 18%), and 17b
(28.8 mg, 23.9%) as white solid.
3.2.11. Benzoylation of 1,2-O-cyclohexylidene-5,6-bis-O-
(o-xylylene phosphoryl)-myo-inositol 12a. Entry 1. To a
CH2Cl2 (2 mL) solution of 12a (62 mg, 0.099 mmol),
pyridine (24.0 mL, 0.298 mmol) and DMAP (cat.) was
added benzoyl chloride (11.6 mL, 0.10 mmol) at 08C for
5.3 h. After being diluted with EtOAc, the solution was
washed with KHSO4 aq., NaHCO3 aq., and brine, the
organic layer was dried, concentrated, and chromatographed
(EtOAc/hexane¼1:1) to afford 15a (33.0 mg, 63%) as white
solid. Physical and spectra data of 15a: Rf¼0.58 (EtOAc/
hexane¼3:1); 1H NMR (400 MHz) d: 8.10 (dd, 2H, J¼1.2,
7.8 Hz, C6H5), 7.58 (t, 1H, J¼7.8 Hz, C6H5), 7.46 (t, 2H,
J¼7.8 Hz, C6H5), 7.18–7.39 (complex, 8H, xylylene H),
5.57–5.67 (complex, 2H, ArCH2O), 5.44 (dd, 1H, J¼3.8,
7.8 Hz, InsH-3), 5.35 (t, 1H, J¼13.4 Hz, ArCH2O), 4.97–
5.25 (complex, ArCH2O, InsH-6), 4.65 (dd, 1H, J¼3.8,
5.4 Hz, InsH-2), 4.56 (2£dd, 1H, J¼7.0, 10.0 Hz, InsH-5),
4.38 (complex, 2H, InsH-1, H-4), 1.76 (complex, 2H,
cyclohexylidene H), 1.58 (complex, 6H, cyclohexylidene
H), 1.28 (br, 2H, cyclohexylidene H); 13C NMR
(100.6 MHz) d: 165.94 (CvO), 135.50, 135.46, 135.26,
135.11, 133.25, 130.05 (2C), 129.64, 129.22 (2C), 129.19,
129.15, 129.13, 129.07, 128.88, 128.79, 128.41, 128.34
(aromatic C), 111.92 (spiral C), 80.7 (t, J¼5.2 Hz), 79.77
(t, J¼6.0 Hz), 77.24 (s), 73.17 (s), 71.44 (s), 70.97 (s)
(InsC-1-C-6), 69.24 (t, 2C, J¼7.7 Hz), 68.82 (d, J¼6.8 Hz),
68.71 (d, J¼6.8 Hz) (ArCH2O), 37.25, 34.88, 24.92, 23.85,
23.71 (cyclohexylidene C); 31P NMR (162 MHz) d: 21.53,
21.88. calcd for C35H38O13P2·1.8H2O: C, 55.23; H, 5.51;
found: C, 55.11; H, 5.39.
1
Compound 17a. Rf¼0.39 ((EtOAc/hexane¼2:1); H NMR
(400 MHz) d: 7.24–7.36 (complex, 8H, aromatic H), 5.69
(dd, 1H, J¼8.2, 13.6 Hz, ArCH2O), 5.60 (dd, 1H, J¼10.0,
13.6 Hz, ArCH2O), 5.31–5.39 (complex, 2H, ArCH2O),
4.93–5.18 (complex, 5H, ArCH2O, InsH-6), 4.36–4.42
(complex, 2H, InsH-2, H-5), 4.26 (dd, 1H, J¼5.8, 7.4 Hz,
InsH-1), 4.11and 4.10 (2£t, 1H, J¼6.9 Hz, InsH-4), 3.94
(dd, 1H, J¼3.6, 6.9 Hz), 3.75 (s, OH-4), 1.80 (complex,
2H, cyclohexylidene H), 1.57–1.64 (complex, 6H, cyclo-
hexylidene H), 1.25 (br, 2H, cyclohexylidene H), 1.00 (t,
9H, J¼7.9 Hz, CH3), 0.67 (q, 6H, J¼7.9 Hz, CH2); 13C
NMR (100.6 MHz) d: 135.60, 138.58, 135.31, 135.29,
129.27, 129.18, 129.15, 129.11, 129.06, 128.87, 128.83
(aromatic C), 111.61 (spiral C), 80.45 (t, J¼4.6 Hz), 80.14
(dd, J¼5.6, 7.3 Hz), 79.62 (t, J¼5.6 Hz), 76.58 (d, J¼
2.0 Hz), 75.34 (s), 71.34 (s) (InsC-1-C-6), 69.30 (d, J¼
7.2 Hz), 69.13 (d, J¼7.2 Hz), 68.81 (d, J¼6.9 Hz), 68.69
(d, J¼6.9 Hz) (ArCH2O), 37.00, 34.73, 25.02, 23.88, 23.72
(cyclohexylidene C), 6.76 (s, CH3), 4.88 (s, CH2); 31P NMR
(162 MHz) d: 21.55, 22.22; calcd for C34H48O12P2Si·H2O:
C, 53.96; H, 6.66; found: C, 53.94; H, 6.47.
1
Compound 17b. Rf¼0.28 (EtOAc/hexane¼2:1); H NMR
(400 MHz) d: 7.31–7.39 (complex, 8H, aromatic H), 5.53
(dd, 1H, J¼10.0, 13.4 Hz, ArCH2O), 5.40 (dd, 1H, J¼10.0,
13.4 Hz, ArCH2O), 5.28 (s, 2H, ArCH2O), 5.24 (s, 2H,
ArCH2O), 5.09 (dd, 1H, J¼13.4, 21.8 Hz, ArCH2O), 4.99
(dd, 1H, J¼13.6, 22.6 Hz, ArCH2O), 4.76 (q, 1H, J¼8.0 Hz,

7710
F. Han et al. / Tetrahedron 59 (2003) 7703–7711
InsH-6), 4.75 (s, 1H, OH-3), 4.60 and 4.58 (2£dd, J¼5.5,
6.3 Hz, InsH-2), 4.40 (dd, 1H, J¼3.7, 6.5 Hz, InsH-4), 4.29
(dd, 1H, J¼6.5, 8.0 Hz, InsH-5), 4.26 (dd, 1H, J¼3.7,
6.3 Hz, InsH-3), 3.93 and 3.92 (2£dd, 1H, J¼5.5, 8.0 Hz,
InsH-1), 1.73 (complex, 2H, cyclohexylidene H), 1.59
(complex, 6H, cyclhexylidene H), 1.27 (complex, 2H,
cyclohexylidene H), 0.95 (t, 9H, J¼7.9 Hz, CH3), 0.62 (q,
6H, J¼7.9 Hz, CH2); 13C NMR (100.6 MHz) d: 135.60,
135.44, 135.42, 135.30, 129.43, 129.36, 129.32, 129.30,
129.25, 129.22, 129.14, 129.04 (aromatic C), 111.25 (spiral
C), 82.79 (d, 1C, J¼5.4 Hz, InsC-5), 81.58 (d, 1C, J¼
5.8 Hz, InsC-6), 75.84 (d, 1C, J¼6.1 Hz, InsC-1), 74.68 (s,
1C, InsC-2), 73.52 (d, 1C, J¼4.3 Hz, InsC-4), 69.25 (d, 1C,
J¼7.7 Hz), 69.18 (d, 1C, J¼4.7 Hz) (ArCH2O), 68.99 (d,
1C, J¼6.9 Hz, InsC-3), 68.73 (t, 2C, J¼6.8 Hz, ArCH2O),
36.86, 34.43, 25.05, 23.96, 23.16 (cyclohexylidene C), 6.72
(CH3), 4.22 (CH2); 31P NMR (162 MHz) d: 21.09, 21.32.
calcd for C34H48O12P2Si·0.5H2O: C, 53.61; H, 6.60; found:
C, 53.52; H, 6.55.
8.8 Hz, InsH-6), 4.74 (br, OH at C-4), 4.52 (dd, 1H, J¼3.7,
5.6 Hz, InsH-2), 4.41 (dt, 1H, J¼7.4, 8.8 Hz, InsH-5), 4.29
(dd, 1H, J¼5.6, 6.4 Hz, InsH-1), 4.14 (dd, 1H, J¼7.4,
7.8 Hz, InsH-4), 3.70 (dd, 1H, J¼3.7, 7.8 Hz, InsH-3), 3.46
(br, OH at C-3), 1.79 (m, 2H, cyclohexylidene), 1.60 (br,
6H, cyclohexylidene), 1.41 (m, 2H, cyclohexylidene); 13C
NMR (100.9 MHz) d: 135.54, 135.44, 135.22, 135.03,
129.32 (2C), 129.27 (2C), 129.18, 129.08, 128.88, 128.81
(aromatic), 111.88 (spiral C), 80.25 (InsC-5), 80.05
(InsC-6), 77.22 (InsC-1), 74.68 (InsC-2), 72.38 (InsC-4),
70.06 (InsC-3), 68.67–69.51 (m, ArCH2O), 37.29, 34.86,
24.92, 23.83–23.66 (3C) (cyclohexylidene); 31P NMR
(162 MHz) d: 20.31, 20.44; Anal. calcd for
C28H34O12P2·2.5H2O: C, 50.23; H, 5.87; Found: C, 50.15;
H, 5.78.
3.2.15. D-2,3-O-Cyclohexylidene-4,5-di-O-(o-xylylene
phosphoryl)-myo-inositol 1-O-[(1,2-O-di-palmitoyl-sn-
glceryl) benzyl] phosphate (D-14a). Phosphorylation of
L-12a with 1,2-di-O-palmitoyl-sn-glycerol phosphite was
proceeded with the general procedures. Physical and
spectral data of D-14a: see Ref. 16.
Entry 6. The reaction was carried out by the same procedure
as in entry 6 at 08C for 4.5 h to afford 17a (21.3%), and 17b
(20.5%).
3.2.16. D-1-O-(1,2-Di-O-palmitoyl-sn-glycerophospho)-
myo-inositol 4,5-bis(dihydrogen phosphate) (1). Pro-
cedure (a). To a CH2Cl2 (2 mL) solution of D-14a
(25.4 mg, 18.9 mmol) was added ethylene glycol (2.7 mL,
47 mmol) and pyridinium poly(hydrogen fluoride) (24 mL,
0.76 mmol) at 08C. The mixture was stirred at the same
temperature for 20 min, and additional 1.2 h at room
temperature. Then saturated aq. NaHCO3 (3 mL) was
added slowly at 08C and stirred for 15 min. After being
diluted with EtOAc, the solution was washed with aq.
KHSO4, aq. NaHCO3, and NaCl, organic layer was dried,
concentrated, and chromatographed on silica gel (EtOAc/
hexane¼1:4) to afford 19 (21.8 mg, 92%). 19 (19.5 mg) thus
obtained was hydrogenated overnight in EtOH/CHCl3
(2 mL, v/v 1:1) in the presence of 10 mg Pd/C, then
100 mL Et3N was added and stirred for 10 min at room
temperature. After filtration to remove the catalyst, the
filtrate was concentrated and dried in vacuum to give 1 as its
triethylammonium salt (19.7 mg, quant.).
3.2.13. O-Xylylene N,N-diethylphospharamidite L-1,2-O-
cyclohexylidene-3,4-O-(tetraisopropyldisiloxane-1,3-
diyl)-5,6-bis-O-(o-xylylene phosphoryl)-myo-inositol
(18). To a CH2Cl2 solution (10 mL) of L-11 (180 mg,
0.36 mmol) and 1-H tetrazole (151 mg, 2.15 mmol) was
added dropwise O-xylylene-N,N-diethyl phospharamidite
(XEPA) (274 mg, 1.15 mmol) at 08C. The mixture was
stirred at room temperature for 1.5 h, and additional 20 min
after 12 mL H2O was added. Then the solution was cooled to
2788C, and treated with m-CPBA (370 mg, 2.15 mmol) for
5 and 30 min at ambient temperature. After being diluted
with EtOAc, the organic layer was washed with 10% aq.
Na2SO3, aq. NaHCO3, and NaCl, dried, and concentrated.
The residue was chromatographed (EtOAc/hexane¼1:5) to
afford 18 (297 mg, 95%) as white solid. Rf¼0.3 (EtOAC/
hexane¼1:2); [a]2D6¼23.9 (c¼3, CHCl3); mp: 82–838C; 1H
NMR (400 MHz) d: 7.22–7.36 (m, 8H), 5.46–5.57 (com-
plex, 2H, ArCH2O), 5.34 (t, 1H, J¼12.8, ArCH2O), 5.04–
5.22 (complex, 5H, ArCH2O), 4.81 (t£2, 1H, J¼8.8 Hz,
InsH-6), 4.57 (t£2, 1H, J¼8.8 Hz, InsH-5), 4.34 (t, 1H,
J¼4.0 Hz, InsH-2), 4.10–4.20 (complex, 2H, InsH-1and
H-4), 4.98 (dd, 1H, J¼4.0, 8.8 Hz, InsH-3), 1.87 (br, 2H,
cyclohexylidene), 1.62 (br, 6H, cyclohexylidene), 1.25 (br,
2H, cyclohexylidene), 1.05–1.12 (complex, 28H, isopro-
pyl); 31P NMR (162 MHz) d: 0.88, 21.31.
Procedure (b). 10%–Pd/C (5 mg) was suspended in a
EtOAc (3 mL) solution of D-14a (20 mg, 15 mmol) and
degassed. The solution was stirred under H2 for 2.5 days at
room temperature, then 100 mL Et3N was added and stirred
for 10 min. The catalyst was removed by filtration, washed
with 5 mL CHCl3, 5 mL MeOH. The combined organic
layer was concentrated and dried in vacuum to give 1 as its
triethylammonium salt (18.8 mg, quant.). Rf¼0.3 (CHCl3/
acetone/MeOH/AcOH/H2O 40/15/15/12/8); [a]2D6¼þ5.41,
3.2.14. L-1,2-O-Cyclohexylidene-5,6-di-O-(o-xylylene
phosphoryl)-myo-inositol (L-12a). To a THF solution
(3 mL) of 18 (298 mg, 0.34 mmol) were added HOAc
(78.7 mL, 1.37 mmol) and TBAF·3H2O (325 mg,
1.03 mmol) at 2208C, and stirred for 12 h. Volatile
materials were evaporated under reduced pressure at
ambient temperature, the residue was chromatographed on
silical gel (EtOAc/hexane¼4:1) to afford L-12a (186 mg,
81%) as white solid. Rf¼0.35 (EtOAc/MeOH 15:1);
[a]2D5¼þ4.3 (c¼1.56, CHCl3); mp decomposed at above
1
(c¼0.61, CHCl3); H NMR (400 MHz) d: 5.21 (br, 1H,
glyceryl sn-2-H), 4.37–4.39 (br, 2H, InsH-2 and H-4), 4.25
(br, 1H, InsH-1), 3.10–4.20 (complex, 2H, glyceryl and
InsH-5), 3.91–4.05 (complex, 4H, InsH-6, glyceryl sn-1 and
2), 3.60 (br, 1H, InsH-3), 3.11 (q, J¼6.8 Hz, NCH2CH3),
2.26 (q, 4H, J¼7.8 Hz, a-H in Pal), 1.57 (br, 4H, b-H
in Pal), 1.38 (t, J¼7.2 Hz, NCH2CH3), 1.25 (br, 48H,
Pal), 0.88 (t, 6H, J¼6.4 Hz, CH3 in Pal); 31P NMR
(162 MHz) d: 3.23, 2.57, 0.41; FAB-MS (triethanolamine,
negative) m/z: 970 [M2H]2, 992 [M22HþNa]2, 1014
[M23Hþ2Na]2.
1
1708C; H NMR (400 MHz) d: 7.32–7.39 (m, 8H), 5.62
(complex, 2H, ArCH2O), 5.31 (t, 1H, J¼13.6 Hz, ArCH2O),
5.01–5.22 (complex, 5H, ArCH2O), 4.91 (dd, 1H, J¼6.4,

F. Han et al. / Tetrahedron 59 (2003) 7703–7711
7711
7407–7410. (b) Toker, A.; Meyer, M.; Reddy, K. K.; Falck,
J. R.; Aneja, R.; Aneja, S.; Parra, A.; Burns, D. J.; Ballas,
L. M.; Cantley, L. C. J. Biol. Chem. 1994, 269, 32358–32367.
(c) Dreef, C. E.; Elie, C. J. J.; Hoogerhout, P.; Marel, G. A. van
der; Boom, J. J. van. Tetrahedron Lett. 1988, 29, 6513–6516.
(d) Krylova, V. N.; Lyutik, A. I.; Gornaeva, N. P.; Shvets, V. I.
Zh. Obshch. Kim. 1981, 51, 210–214, (translated version in
English, 183–186).
3.2.17. D-2,3-O-Cyclohexylidene-1-O-(dibenzyl pho-
phoryl)-5,6-di-O-(o-xylylene phosphoryl)-myo-inositol
(D-14e). Phosphorylation of L-12a with tribenzyl phos-
phite was proceeded with the general procedures to afford
D-14e. [a]2D6¼þ2.33, (c¼1.4, CHCl3); spectral data were
consistent with racemic form 14e.
3.2.18. D-myo-Inositol 1,4,5-trisphosphate (2). Procedure
(a). 10%–Pd/C (10 mg) was suspended in a EtOH (2 mL)
solution of D-14e (16.3 mg, 18.4 mmol) and degassed. The
solution was stirred under H2 for 4 h at room temperature.
The catalyst was removed by filtration, washed with 10 mL
EtOH. The combined organic layer was evaporated to
dryness under reduced pressure. The residue was dissolved
in a 1:2 (v/v) mixture of H2O and HOAc, stirred for 44 h at
room temperature. Volatile materials was evaporated under
reduced pressure, then dried in vacuum for 5 h. The residue
was passed through a Naþ-form of cation-resin using H2O as
mobile phase to afford trisodium salt of 1 (8.8 mg, quant.).
8. PtdIns(4,5)P2 from L-(2)-quebrachitol: Qiao, L.; Hu, Y.; Nan,
F.; Powis, G.; Kozikowski, A. P. Org. Lett. 2000, 2(2),
115–117.
9. Potter, B. V. l.; Lampe, D. Angew. Chem., Int. Ed. Engl. 1995,
34, 1933–1972, and references therein.
10. Garrett, S. M.; Liu, C.; Riley, A. M.; Potter, B. V. L. J. Chem.
Soc., Perkin Trans. 1 1998, 1367–1368.
11. (a) Ling, L.; Ozaki, S. Carbohydr. Res. 1994, 256, 49–58.
(b) Ling, L.; Ozaki, S. Tetrahedron Lett. 1993, 34,
2501–2504.
12. (a) Watanabe, Y.; Mitani, M.; Morita, T.; Ozaki, S. J. Chem.
Soc., Chem. Commun. 1989, 482–483. (b) Watanabe, Y.;
Ishikawa, H. Tetrahedron Lett. 2000, 8509, 8512.
13. Watanabe, Y.; Abe, Y.; Takao, H. Carbohydr. Lett. 1998, 3,
85–90.
Procedure (b). To 8 mL mixed solvent of MeOH/CHCl3
(v/v 1:1) was added D-14e (30 mg, 34 mmol), tributylamine
(16.1 mL, 68 mmol) and 10 mg 10%-Pd/C, the solution was
stirred under H2 at room temperature for 3 days. The
catalyst was removed by filtration, washed with 10 mixed
solvent of MeOH/CHCl3 (v/v 1:1), 10 mL MeOH. The
combined organic layer was evaporated to dryness under
reduced pressure, the residure was passed successively
through cation-exchange column of Hþ and Naþ-form to
give 1 as its trisodium salt quantitatively. [a]2D6¼28.70,
(c¼0.46, H2O); spectral data were consistent with those of
the specimen prepared formerly by us.28
14. (a) Watanabe, Y.; Hirofuji, H.; Ozaki, S. Tetrahedron Lett.
1994, 35, 123–124. (b) Watanabe, Y.; Tomioka, M.; Ozaki, S.
Tetrahedron 1995, 51, 8969–8976. (c) Watanabe, Y.;
Nakatomi, M. Tetrahedron Lett. 1998, 39, 1583–1586.
15. (a) Watanabe, Y.; Yamamoto, T.; Ozaki, S. J. Org. Chem.
1996, 61, 14–15. (b) Watanabe, Y.; Yamamoto, T.; Okazaki,
T. Tetrahedron 1997, 53, 903–918.
16. Han, F. S.; Hayashi, M.; Watanabe, Y. Chem. Lett. 2003, 32,
46–47.
17. (a) Reddy, K. K.; Rizo, J.; Falck, J. R. Tetrahedron Lett. 1997,
38, 4729–4730. (b) Falck, J. R.; Krishna, U. M.; Katipally,
K. R.; Capdevila, J. H.; Ulug, E. T. Tetrahedron Lett. 2000, 41,
4271–4275.
Acknowledgements
We are grateful to the Center for Cooperative Research and
Development of Ehime University for MS analysis.
18. Watanabe, Y.; Shinohara, T.; Fujimoto, T.; Ozaki, S. Chem.
Pharm. Bull. 1990, 38, 562–563.
19. (a) Studies in Natural Products Chemistry; Rahman, A., Ed.;
Elsevier: Amsterdam, 1996; Vol. 18, pp 391–456 Part K.
(b) Recent report concerning the selective acylation: Nicholas,
G. M.; Covac, P.; Bewley, C. A. J. Am. Chem. Soc. 2002, 124,
3492–3493.
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