3-(Hydroxymethyl)-bearing phosphatidylinositol ether lipid analogues and carbonate surrogates block PI3-K, akt, and cancer cell growth

Article abstract of DOI:10.1021/jm000117y

Phosphatidylinositol 3-kinase (PI3-K) phosphorylates the 3-position of phosphatidylinositol to give rise to three signaling phospholipids. Binding of the pleckstrin homolgy (PH) domain of Akt to membrane PI(3)P's causes the translocation of Akt to the plasma membrane bringing it into contact with membrane-bound Akt kinase (PDK1 and 2), which phosphorylates and activates Akt. Akt inhibits apoptosis by phosphorylating Bad, thus promoting its binding to and blockade of the activity of the cell survival factor Bcl-x. Herein we present the synthesis and biological activity of several novel phosphatidylinositol analogues and demonstrate the ability of the carbonate group to function as a surrogate for the phosphate moiety. Due to a combination of their PI3-K and Akt inhibitory activities, the PI analogues 2, 3, and 5 proved to be good inhibitors of the growth of various cancer cell lines with IC₅₀ values in the 1-10μM range. The enhanced Akt inhibitory activity of the axial hydroxymethyl-bearing analogue 5 compared to its equatorial counterpart 6 is rationalized based upon postulated differences in the H-bonding patterns of these compounds in complex with a homology modeling generated structure of the PH domain of Akt. This work represents the first attempt to examine the effects of 3-modified PI analogues on these two crucial cell signaling proteins, PI3-K and Akt, in an effort to better understand their cell growth inhibitory properties.

Full text of DOI:10.1021/jm000117y

J . Med. Chem. 2000, 43, 3045-3051
3045
3-(Hyd r oxym eth yl)-Bea r in g P h osp h a tid ylin ositol Eth er Lip id An a logu es a n d
Ca r bon a te Su r r oga tes Block P I3-K, Ak t, a n d Ca n cer Cell Gr ow th
Youhong Hu,† Lixin Qiao,† Shaomeng Wang,† Suo-bao Rong,† Emmanuelle J . Meuillet,‡ Margareta Berggren,‡
Alfred Gallegos,‡ Garth Powis,‡ and Alan P. Kozikowski*,†
Drug Discovery Program, Department of Neurology, Georgetown University Medical Center, 3970 Reservoir Road NW,
Washington, D.C. 20007, and Arizona Cancer Center, 1515 North Campbell Avenue, Tucson, Arizona 85724
Received March 10, 2000
Phosphatidylinositol 3-kinase (PI3-K) phosphorylates the 3-position of phosphatidylinositol to
give rise to three signaling phospholipids. Binding of the pleckstrin homolgy (PH) domain of
Akt to membrane PI(3)P’s causes the translocation of Akt to the plasma membrane bringing
it into contact with membrane-bound Akt kinase (PDK1 and 2), which phosphorylates and
activates Akt. Akt inhibits apoptosis by phosphorylating Bad, thus promoting its binding to
and blockade of the activity of the cell survival factor Bcl-x. Herein we present the synthesis
and biological activity of several novel phosphatidylinositol analogues and demonstrate the
ability of the carbonate group to function as a surrogate for the phosphate moiety. Due to a
combination of their PI3-K and Akt inhibitory activities, the PI analogues 2, 3, and 5 proved
to be good inhibitors of the growth of various cancer cell lines with IC50 values in the 1-10 µM
range. The enhanced Akt inhibitory activity of the axial hydroxymethyl-bearing analogue 5
compared to its equatorial counterpart 6 is rationalized based upon postulated differences in
the H-bonding patterns of these compounds in complex with a homology modeling generated
structure of the PH domain of Akt. This work represents the first attempt to examine the
effects of 3-modified PI analogues on these two crucial cell signaling proteins, PI3-K and Akt,
in an effort to better understand their cell growth inhibitory properties.
In tr od u ction
cer.3 An important counterpart to PI3-K is the tumor
suppressor PTEN, a protein that is able to bring about
the dephosphorylation of PI(3,4,5)P3, with specificity
being shown for the phosphate at the D-3 position of
the inositol ring. Mutations in the PTEN tumor sup-
pressor gene appear to be a common occurrence in a
number of human cancers.4 Thus, PI3-K and Akt
provide novel targets for drugs to inhibit the repression
of apoptosis in cancer cells and thereby the opportunity
to overcome the effects of the loss of the tumor supressor
PTEN.
Our earlier work has shown that certain D-3-deoxy-
substituted myo-inositols are taken up by the myo-
inositol transporter of cells and incorporated into cel-
lular PI’s by PI synthetase leading to selective cell
growth inhibition of some transformed cells.5 However,
the activity of these compounds is inhibited by physi-
ological concentrations of the natural substrate, myo-
inositol. To overcome the transport and synthesis prob-
lem, we were led to investigate the activity of certain
3-modified PI analogues. The 3-deoxy PI ether lipid 1
(DPIEL), for example, was found to inhibit PI3-K with
an IC50 of 14.8 ( 5.6 µM and to block the growth of HT-
29 colon cancer cells with an IC50 of 2.1 µM. This
compound was also found to inhibit the growth of
MCF-7 human breast cancer xenografts in scid mice
(T/C < 16%) when administered ip as a micellar
suspension daily for 17 days at 400 mg/kg.6 DPIEL is
presently under development by the RAID program of
the National Cancer Institute (NCI). Because of the
interesting biology shown by DPIEL, we deemed it
valuable to investigate the activity of other 3-modified
PI ether lipid analogues with the aim to further improve
Phosphatidylinositol 3-kinase (PI3-K) phosphorylates
the 3-position of phosphatidylinositol (PI), PI(4)P, and
PI(4,5)P2 to give rise to three signaling phospholipids:
PI(3)P, PI(3,4)P2, and PI(3,4,5)P3, respectively.1 These
3-phosphorylated PI’s have the unique ability to bind
to specific protein domains, the so-called pleckstrin
homology (PH) domains, of a number of signaling
proteins. One of the most extensively studied of the PH
domain-regulated signaling proteins acting downstream
of PI3-K is the proto-oncogenic serine/threonine kinase
Akt [also known as RAC-PK or protein kinase B (PKB)].
In particular, while the PH domain of Akt binds both
PI(3,4)P2 and PI(3,4,5)P3 in vitro, only PI(3,4)P2 acti-
vates Akt. Binding of the PH domain of Akt to mem-
brane PI(3)P’s causes the translocation of Akt to the
plasma membrane bringing it into contact with mem-
brane-bound Akt kinases [phosphatidylinositol-depend-
ent kinase-1 and -2 (PDK1 and 2)], which phosphorylate
and activate Akt. Akt is a proto-oncogene that inhibits
apoptosis by phosphorylating a number of downstream
targets. This includes phosphorylation of Bad, which
promotes its binding to and blockade of the activity of
the cell survival factor Bcl-x.2 Thus, the inhibition of
Akt activation induces cancer cell apoptosis. Three
mammalian isoforms of Akt have been identified: Akt1,
Akt2, and Akt3. Akt1 has been found to be overex-
pressed in gastric adenocarcinomas, while Akt2 is
overexpressed in breast, ovarian, and pancreatic can-
* To whom correspondence should be addressed. Tel: 202-687-0686.
Fax: 202-687-5065. E-mail: kozikowa@giccs.georgetown.edu.
† Georgetown University Medical Center.
‡ Arizona Cancer Center.
10.1021/jm000117y CCC: $19.00 © 2000 American Chemical Society
Published on Web 07/18/2000

3046 J ournal of Medicinal Chemistry, 2000, Vol. 43, No. 16
Hu et al.
Ch a r t 1
Sch em e 1a
a
Reagents and conditions: (a) (i) DMSO, (COCl)2, CH2Cl2, -78
°C, (ii) MePPh3Br, n-BuLi, THF, -78 °C to rt; (b) 9-BBN, NaOH,
H2O2, THF, 0-50 °C; (c) (i) (S)-(-)-camphanic chloride, Et3N,
CH2Cl2, rt, (ii) 1 N HCl, MeOH, rt.
upon Akt inhibition and antiproliferative action. In the
present article we report on the synthesis and biology
of analogues that bear a hydroxymethyl group at
position 3 and which also contain in two cases a second
modification in which a carbonate group has been
introduced as a surrogate for the phosphate linkage
between the inositol ring and ether lipid.
Sch em e 2a
Ch em istr y
The starting material 7 was obtained from L-quebra-
chitol as reported previously.7 Swern oxidation of the
free axial hydroxyl group gave an unstable ketone that
was transformed into alkene 8 through a Wittig reac-
tion.8 Hydroboration with 9-BBN followed by oxidation
with alkaline hydrogen peroxide solution converted 8
into alcohols 9a ,b (approximately 1:1) in 90% yield after
chromatography. Compounds 9a ,b were readily distin-
a
Reagents and conditions: (a) (i) NaH, BnBr, DMF, 0 °C to rt,
(ii) AcCl (cat.), CH2Cl2-MeOH, rt; (b) (i) NaH, BnBr, DMF, 0 °C
to rt, (ii) concd HCl, MeOH, rt; (c) (i) Bu2SnO, toluene, reflux, (ii)
allyl-Br, CsF, -50 °C to rt, (iii) NaH, BnBr, DMF, 0 °C to rt; (d)
(i) RhCl(PPh3)3 (cat.), DABCO, EtOH, reflux, (ii) 1 N HCl, acetone,
reflux; (e) (i) BnOP(Ni-Pr2)2, diisopropylammonium tetrazolide,
CH2Cl2, rt, (ii) 2-O-methyl-1-O-octadecyl-sn-glycerol, tetrazole,
CH2Cl2, rt, (iii) t-BuOOH, CH2Cl2, rt; (f) H2, 20% Pd(OH)2/C,
t-BuOH, rt.
1
guished through the H NMR signals of the protons at
C-3. Furthermore, we confirmed the stereochemistry of
9a by X-ray crystal structure analysis of its (1S)-(-)-
camphanate ester 10 (Figure 1). The availability of both
intermediates 9a ,b allowed us to investigate the effect
of C-3 stereochemistry on activity (Scheme 1).
Sch em e 3a
Following similar procedures as published before,5
compounds 9a ,b were protected with a benzyl group,
and the trans-acetonide moieties were then removed by
controlled acidic hydrolysis to give compounds 11a ,b.
Both of the resulting hydroxyl groups were again
benzylated, and the remaining cis-acetonide was re-
moved by acidic hydrolysis. The compounds 12a ,b were
selectively allylated at position 1 via a 1,2-O-stannylene
intermediate. After benzylation at position 2, isomer-
ization of the double bond in the allyl group with RhCl-
(PPh3)3 and DABCO in ethanol and subsequent acidic
hydrolysis furnished the desired intermediates 14a ,b.
Installation of the phospholipid side chain and depro-
tection were performed in a similar manner as in the
synthesis of 1 (Scheme 2).
a
Reagents and conditions: (a) 1,1′-carbonyldiimidazole, toluene,
reflux; (b) DBU, 2-O-methyl-1-O-octadecyl- sn-glycerol, toluene,
reflux; (c) H2, 20% Pd(OH)2/C, t-BuOH, rt.
nyldiimidazole to form the carbamates 16a ,b. The
second imidazolyl group was then replaced by the ether
lipid by refluxing the reactants in toluene in the
presence of DBU to give compounds 17a ,b. Final hy-
drogenolysis then delivered the desired carbonate ana-
logues 5 and 6 in good purity (Scheme 3). Carbonate 4
was prepared in a similar fashion using the appropri-
ately protected 3-deoxyinositol.
To prepare the carbonate analogues 5 and 6, com-
pounds 14a ,b were refluxed in toluene with 1,1-carbo-
Biologica l Activity
The five new PI analogues were tested for their ability
to inhibit p100/p85 PI3-K using the methods described
previously.5c To measure the effects of these compounds
on Akt activity, cells were grown in 6-well plates in
Dulbecco’s modified Eagle medium (DMEM) supple-
mented with 10% heat-inactivated fetal bovine serum
F igu r e 1. Single-crystal X-ray structure of the camphanate
ester 10.

Phosphatidylinositols and Carbonate Surrogates
J ournal of Medicinal Chemistry, 2000, Vol. 43, No. 16 3047
(FBS), 4.5 g/L glucose, 100 U/mL penicillin, and 100 µg/
mL streptomycin in a 5% CO2 atmosphere. Before the
experiment, the medium was replaced by DMEM with-
out FBS for 16 h. The cells were incubated with the
different drugs for 3 h at 37 °C. The concentrations
tested ranged from 0 to 20 µM. Monolayers were then
stimulated with PDGF (50 ng/mL) for another hour at
37 °C. Control cells were incubated with DMEM without
PDGF. After the stimulation, the culture medium was
aspirated, and cells were lyzed in Laemmli buffer.9
Lysates were boiled for 5 min, and an equivalent
amount of protein was loaded on a 7.5% SDS-PAGE.
Proteins were electrophoretically transferred to PVDF
membranes, preincubated in blocking buffer (phosphate-
buffered saline containing 5% bovine serum albumin),
and incubated with anti-phospho-Akt polyclonal anti-
bodies (New England Biolabs). Immunoreactive bands
were detected using an anti-rabbit antibody coupled to
horseradish peroxidase and the Renaissance chemlu-
minescence kit (NEN). Phosphorylation of Akt was
quantified using Eagle Eye software (Biorad) and Kodak
film.
F igu r e 2. (a) Ribbon diagram of the PH domain of Akt in
complex with the phosphorylation product of 5. The â-strands,
R-helix, turn, and coils are colored yellow, red, blue, and green,
respectively. Homology modeling was done using the homology
package of InsightII (Molecular Simulation, Inc., San Diego,
CA) with the crystal structure of the Btk PH domain (PDB
code: 1BWN) as the template. (b) Model of 5 in complex with
the PH domain of Akt. (c) Model of 6 in complex with the PH
domain of Akt. Hydrogen bonds are depicted in the figures.
Docking was performed using the program FlexX.11
For the cell growth inhibition studies MCF-7 (breast),
HT-29 (colon), Hela (cervical), and PC-3 (prostate) cells
were seeded into 12-well plates in DMEM supplemented
with 10% FBS; 30 000 cells/well were plated, and after
16 h the drugs were added to the wells. After 3 days in
DMEM/10% FBS in the absence or presence of different
concentration of the drugs, cells were counted using a
hemocytometer.
Ta ble 1. IC50 Values for Inhibition of Akt and PI3-K Activity
by Compounds 1-6
Mod elin g
IC50 (µM)
Akt PI3-K
IC50 (µM)
Akt PI3-K
The 3-modified inositol analogues in complex with the
PH domain of Akt have been modeled using homology
modeling and docking methods.10,11 In cells, the PI
analogues are likely to have undergone some level of
phosphorylation at positions 4 and 5, and therefore we
chose to use the phosphorylated analogues in the
docking studies.12 The complex models generated for the
4,5-bisphosphorylated derivatives of 5 and 6 are shown
in Figure 2 and are further discussed in the following
section.
compd
compd
1
2
3
1.5 ( 0.3 14.8 ( 5.6
7.8 ( 0.8 31.0 ( 7.0
9.1 ( 1.7 18.5 ( 1.7
4
5
6
12.5 ( 2.0 15.5 ( 1.8
5.0 ( 1.7 83.0 ( 21.0
32.0 ( 3.5 21.3 ( 7.6
6-fold reduction in Akt inhibitory activity compared to
5. In terms of PI3-K inhibition, the equatorial carbonate
6 is more active than its axial counterpart 5; this same
trend is observed in comparing the phosphate analogues
3 versus 2. In general, both carbonates are relatively
poor inhibitors of PI3-K, which suggests the importance
of the phosphate moiety to recognition by this enzyme.
The modeling results show that these 4,5-phospho-
rylated PI’s interact in a complementary fashion with
the positively charged pocket formed by the â1-â2 and
â3-â4 loops of the PH domain of Akt (Figure 2a). The
axial hydroxymethyl group in 5 is anchored through
hydrogen bonds with Arg25 (Figure 2b), whereas the
equatorial hydroxymethyl group in 6 is improperly
situated to interact with Arg25, thus resulting in the
loss of this H-bond interaction, as well as one additional
H-bond between the headgroup and Arg23 (Figure 2c).
The differences in the binding interactions may be used
in a qualitative fashion to rationalize the differences in
the activities of 5 and 6 for Akt inhibition. Further
efforts to evaluate the accuracy of this model by utilizing
it in the design of analogues of possibly improved Akt
inhibitory activity are being made.
Resu lts a n d Discu ssion
Data for the five new PI analogues from the PI3-K
and Akt activity studies are presented in Table 1 along
with comparison data obtained previously for DPIEL
(1). It is interesting to note that the PI analogues
bearing a hydroxymethyl group at position 3 all exhibit
some activity for the inhibition of Akt and PI3-K. The
phosphate-containing analogues 2 and 3 show compa-
rable Akt inhibitory activity, while 3 is slightly more
potent than 2 in the inhibition of PI3-K. Among these
newly synthesized compounds, the axial hydroxymethyl-
bearing PI analogue 5 is the best inhibitor of Akt with
an IC50 of 5.0 ( 1.9 µM, although it is about 3-fold less
potent than 1. Nevertheless, it is also important to
notice that 5 is more specific than DPIEL for Akt
inhibition. DPIEL exhibits an IC50 of 14.8 ( 5.6 µM as
compared to 83.0 ( 21.0 µM for compound 5 in the
inhibition of PI3-K. In fact, among all of these ana-
logues, carbonate 5 is the least potent for inhibition of
PI3-K. Positioning of the hydroxymethyl group in the
equatorial orientation as in the carbonate 6 leads to a
In terms of inhibition of cell growth of the various cell
lines tested, the PI analogues 2, 3, and 5 generally
proved to be slightly more potent than the others, with

3048 J ournal of Medicinal Chemistry, 2000, Vol. 43, No. 16
Hu et al.
Ta ble 2. Effects of Compounds 1-6 on the Growth Inhibition
of Cancer Cell Lines in Vitro
tography on silica gel (EtOAc/hexanes 1:10) to give the product
(850 mg, 54%): mp 110.5 °C; [R]D -63.8° (c 0.77, CHCl3); H
1
NMR δ 7.43-7.24 (m, 5H), 5.40 (d, 1H, J ) 0.9 Hz), 5.31 (s,
1H), 4.84 (s, 2H), 4.64 (d, 1H, J ) 5.7 Hz), 4.25 (d, 1H, J ) 9.6
Hz), 4.19 (t, 1H, J ) 6.0 Hz), 3.75 (dd, 1H, J ) 10.5, 6.3 Hz),
3.39 (t, 1H, J ) 10.2 Hz), 1.51 (s, 3H), 1.46 (s, 3H), 1.39 (s,
3H), 1.38 (s, 3H); 13C NMR δ 138.28, 138.22, 128.16, 127.95,
127.43, 113.40, 112.19, 110.02, 81.97, 81.68, 80.89, 79.34,
76.12, 72.96, 27.75, 27.22, 26.91, 25.92.
IC50 (µM)
compd
HT-29
MCF-7
Hela
PC-3
1
2
3
4
5
6
2.1
4.5
7.5
2.5
10.0
12.0
7.2
5.0
2.0
12.0
1.2
12.5
7.0
1.0
7.5
ND
4.0
2.0
7.0
2.0
30.0
2.5
>20.0
1L-3-O-Ben zyl-6-d eoxy-6-(h yd r oxym eth yl)-1,2:4,5-d i-O-
isop r op ylid en e-ch ir o-in ositol (9a ) a n d 1D-6-O-Ben zyl-3-
d eoxy-3-(h yd r oxym et h yl)-1,2:4,5-d i-O-isop r op ylid en e-
m yo-in ositol (9b). To a solution of compound 8 (1.80 g, 5.2
mmol) in anhydrous THF (20 mL) was added 9-BBN (2.0 g,
8.2 mmol) at room temperature under N2. The mixture was
stirred at room temperature for 12 h and at 50 °C for 2 h,
then cooled in an ice bath. Ethanol (8 mL), 3 M NaOH (4 mL)
and 35% H2O2 (4 mL) were added dropwise. The mixture was
warmed to 50 °C for 1 h and evaporated, and H2O (10 mL)
was added. The product was extracted into CH2Cl2 (3 × 100
mL) and the organic phase was dried over MgSO4 and
evaporated. The residue was purified by column chromatog-
raphy on silica gel (EtOAc/hexanes 1:2) to give 9a (910 mg,
48%) and 9b (790 mg, 42%). Compound 9a : mp 113-114 °C;
10.0
IC50 values in the 1-10 µM range (Table 2). It is likely
that the cell growth inhibitory effects of these com-
pounds are due to a combination of their PI3-K and Akt
inhibitory activities, in addition to nonspecific effects
on membrane structure. However, upon the basis of the
present data set, growth inhibition appears to correlate
best with the inhibition of the downstream target Akt.
To the best of our knowledge, this work represents the
first attempt to examine the effects of 3-modified PI
analogues on these two crucial cell signaling proteins,
in an effort to better understand their cell growth
inhibitory properties.
1
[R]D -103.5° (c 0.68, CHCl3); H NMR δ 7.42-7.23 (m, 5H),
4.83 (s, 2H), 4.22 (d, 1H, J ) 5.4 Hz), 4.12 (t, 1H, J ) 6.0 Hz),
3.97 (dd, 1H, J ) 10.2, 5.7 Hz), 3.91 (m, 1H), 3.85 (t, 1H, J )
9.9 Hz), 3.73 (m, 1H), 3.62 (dd, 1H, J ) 9.9, 6.6 Hz), 2.85 (dd,
1H, J ) 12.9, 5.4 Hz), 2.46 (d, 1H, J ) 8.1 Hz), 1.46 (s, 3H),
1.45 (s, 3H), 1.35 (s, 3H), 1.34 (s, 3H); 13C NMR δ 138.19,
128.17, 127.97, 127.45, 111.24, 108.52, 80.99, 80.61, 77.01,
76.66, 76.17, 72.09. 60.73, 39.36, 27.77, 26.99, 26.77, 25.81.
Anal. (C20H28O6) C, H. Compound 9b: mp 117 °C; [R]D -63.5°
(c 1.2, CHCl3); 1H NMR δ 7.42-7.23 (m, 5H), 4.83 (s, 2H), 4.42
(t, 1H, J ) 4.5 Hz), 4.16 (t, 1H, J ) 6.3 Hz), 3.97 (dd, 1H, J )
11.4, 5.4 Hz), 3.89 (m, 1H), 3.76 (dd, 1H, J ) 10.8, 9.0 Hz),
3.63 (dd, 1H, J ) 10.5, 6.6 Hz), 3.46 (t, 1H, J ) 9.6 Hz), 2.28
(m, 1H), 2.14 (m, 1H), 1.46 (s, 3H), 1.43 (s, 3H), 1.33 (s, 6H);
13C NMR δ 138.22, 128.16, 127.96, 127.43, 111.99, 109.63,
106.57, 81.67, 81.37, 80.35, 77.06, 74.97, 71.97, 62.22, 41.67,
27.84, 27.08, 27.03, 25.92. Anal. (C20H28O6) C, H.
In summary, we present the synthesis and biological
activity of several novel phosphatidylinositol analogues
and demonstrate the ability of the carbonate group to
function as a surrogate for the phosphate moiety. The
stereochemistry of the hydroxymethyl substituent at the
3-position is also shown to play some role in Akt and
PI3-K inhibition. Of general interest is the finding that
while deletion of the 3-hydroxy group from the inositol
ring as in DPIEL leads to an antiproliferative agent,
this activity is retained by an inositol homologue modi-
fied by insertion of a single carbon atom between the
cyclohexane ring and the 3-OH. The present findings
in concert with results gleaned from the modeling
studies are being used to further improve compound
potency and to simplify analogue design.
1L -3-O-Be n zyl-6-d e oxy-6-[(1′-(S )-(ca m p h a n oyloxy)-
m eth yl]-ch ir o-in ositol (10). To a solution of 9a (50 mg, 0.13
mmol) in CH2Cl2 (2 mL) were added, under N2, Et3N (130 µL,
0.72 mmol), DMAP (10 mg), and (1S)-(-)-camphanic chloride
(61 mg, 0.28 mmol). The mixture was then stirred for 18 h at
room temperature. CH2Cl2 (50 mL) was added and the solution
was washed with 5% HCl (10 mL), aqueous NaHCO3 (10 mL),
and brine (10 mL). The organic phase was dried over MgSO4
and evaporated. The residue was purified by column chroma-
tography on silica gel (EtOAc/hexane 1:2) to give the ester:
1H NMR δ 7.35 (m, 5H), 4.83 (s, 2H), 4.53 (dd, 1H, J ) 4.2,
11.7 Hz), 4.34 (d, 1H, J ) 5.4 Hz), 4.29-4.20 (m, 2H), 3.94
(dd, 1H, J ) 5.7, 9.9 Hz), 3.69-3.60 (br m, 2H), 2.93 (m, 1H),
2.43 (m, 1H), 2.09-1.87 (br m, 2H), 1.74-1.62 (br m, 2H), 1.46
(s, 3H), 1.40 (s, 3H), 1.36 (s, 3H), 1.33 (s, 3H), 1.12 (s, 3H),
1.06 (s, 3H), 0.97 (s, 3H).
Exp er im en ta l Section
Gen er a l Meth od s. NMR spectra were acquired at a proton
frequency of 300 MHz, using CDCl3 as solvent unless noted
otherwise. 1H chemical shifts are reported with Me4Si (δ )
0.00 ppm) or CHCl3 (δ ) 7.26 ppm) as internal standards. 31
P
chemical shifts are relative to external aqueous 85% H3PO4
(δ ) 0.00 ppm), and 13C chemical shifts as reported with CDCl3
(δ ) 77.00 ppm) or TMS (δ ) 0.00 ppm) as internal standards.
Optical rotations were measured at room temperature. 1-O-
Octadecyl-2-O-Me-sn-glycerol was prepared from (S)-(+)-2,2-
dimethyl-1,3-dioxolane-4-methanol by a reported procedure.6b
1D-6-O-Ben zyl-3-d eoxy-1,2:4,5-d i-O-isop r op ylid en e-3-
m eth ylen e-m yo-in ositol (8). To a solution of (COCl)2 (0.50
mL, 5.50 mmol) in dry CH2Cl2 (5 mL) was added DMSO (0.8
mL, 9.30 mmol) in CH2Cl2 (3 mL) at -78 °C. After 15 min, a
solution of 7 (1.60 g, 4.57 mmol) in CH2Cl2 (10 mL) was added
dropwise at -78 °C, and stirring was continued for an
additional 2 h. EtNi-Pr2 (8 mL) was added, and the mixture
was warmed slowly to room temperature. The organic phase
was washed with H2O (4 × 50 mL) and brine (50 mL) and
dried over MgSO4. The solvent was evaporated, and the crude
product was used for the next step without purification.
To a well-stirred suspesion of CH3(PPh3)3Br (4.20 g, 11.6
mmol) in THF (20 mL) was added dropwise n-BuLi (5 mL, 11.6
mmol) at 0 °C. After 1 h, a solution of the crude ketone in
THF (20 mL) was added at -78 °C, and the mixture was
allowed to warm to room temperature overnight. Et2O (100
mL) was added, and the organic phase was washed with H2O
(50 mL) and brine (50 mL) and dried over MgSO4. After
concentration, the residue was purified by column chroma-
To a solution of the ester in MeOH (2 mL) was added
concentrated HCl (2 drops) at room temperature, and the
solution was stirred for 2 h. The solvent was evaporated, and
the residue was crystallized from EtOAc/EtOH (5 mL, 4:1).
The resulting crystalline solid was analyzed by X-ray diffrac-
tion.
1L-3-O-Ben zyl-6-d eoxy-6-(ben zyloxym eth yl)-1,2-O-iso-
p r op ylid en e-ch ir o-in ositol (11a ) a n d 1D-6-O-Ben zyl-3-
d eoxy-3-(b en zyloxym et h yl)-1,2-O-isop r op ylid en e-m yo-
in ositol (11b). To a suspension of NaH (56 mg, 1.41 mmol)
in DMF (5 mL) was added a solution of compound 9a (343 mg,
0.94 mmol) in DMF (5 mL) at 0 °C. After 30 min, BnBr (0.16
mL, 1.41 mmol) was added dropwise. The mixture was stirred
overnight, then poured into ice water and extracted with
EtOAc (3 × 50 mL). The organic phase was washed with H2O

Phosphatidylinositols and Carbonate Surrogates
J ournal of Medicinal Chemistry, 2000, Vol. 43, No. 16 3049
(50 mL) and brine (50 mL) and dried over MgSO4. The solvent
was evaporated and the residue was used direactly in the next
step.
1H NMR δ 7.31 (m, 20H), 5.91 (m, 1H), 5.26 (dd, 1H, J ) 17.1,
1.5 Hz), 5.17 (d, 1H, J ) 10.2 Hz), 4.85, 4.77 (ABq, 2H, J )
10.8 Hz), 4.80 (s, 2H), 4.63 (s, 2H), 4.53, 4.46 (ABq, 2H, J )
12.3 Hz), 4.26 (s, 1H), 4.14 (d, 2H, J ) 5.4 Hz), 4.04 (dd, 1H,
J ) 9.3, 6.3 Hz), 3.81-3.66 (m, 4H), 3.53 (t, 1H, J ) 8.4 Hz),
2.67 (m, 1H), 2.56 (s, 1H); 13C NMR δ 138.95, 138.86, 138.63,
138.18, 134.74, 128.40, 128.31, 128.29, 128.27, 128.03, 127.92,
127.64, 127.61, 127.50, 127.42, 117.25, 82.79, 81.75, 80.20,
78.38, 75.83, 75.52, 73.13, 72.78, 71.71, 68.44, 66.39. 41.44.
Anal. (C38H42O6) C, H. 1D-1-Allyl-4,5,6-O-Ben zyl-3-d eoxy-
3-(ben zyloxym eth yl)-m yo-in ositol was prepared in the
same manner in 77% yield: 1H NMR δ 7.33 (m, 20H), 6.01
(m, 1H), 5.36 (dd, 1H, J ) 17.4, 1.2 Hz), 5.24 (d, 1H, J ) 10.5
Hz), 4.99-4.86 (m, 5H), 4.56 (d, 2H), 4.52 (s, 1H), 4.38 (s, 1H),
4.26 (m, 2H), 4.02 (t, 1H, J ) 9.3 Hz), 3.99-3.79 (m, 3H), 3.60
(t, 1H, J ) 9.3 Hz), 3.40 (dd, 1H, J ) 9.6, 2.7 Hz), 3.09 (s, 1H),
1.81 (m, 1H).
To a solution of the above crude product in CH2Cl2/MeOH
(12 mL, 5:1) was added acetyl chloride (3 drops). The mixture
was stirred under close TLC control for 20 min, then the
reaction was quenched with Et3N (100 µL). After evaporation,
the residue was purified by column chromatography on silica
gel (EtOAc/hexane 1:2) to give product (278 mg, 72%): 1H
NMR δ 7.32 (m, 10H), 4.92, 4.62 (ABq, 2H, J ) 11.7 Hz), 4.49,
4.43 (ABq, 2H, J ) 12.0 Hz), 4.27 (m, 2H), 3.94 (dd, 1H, J )
7.8, 5.4 Hz), 3.75 (m, 2H), 3.60 (dd, 1H, J ) 9.3, 4.8 Hz), 3.44
(t, 1H, J ) 7.2 Hz), 3.32 (s, 1H), 3.07 (s, 1H), 2.46 (m, 1H),
1.46 (s, 3H), 1.34 (s, 3H); 13C NMR δ 138.32, 137.75, 128.31,
128.24, 127.94, 127.57, 127.54, 127.38, 108.33, 83.24, 79.50,
75.59, 73.22, 72.89, 70.37, 67.21, 40.39, 27.97, 25.84. Anal.
(C24H30O6) C, H. Compound 11b was prepared in the same
manner in 64% yield: mp 66-67 °C; [R]D +2.8° (c 1.4, CHCl3);
1H NMR δ 7.34 (br m, 10H), 4.98, 4.69 (ABq, 2H, J ) 11.7
Hz), 4.57, 4.52 (ABq, 2H, J ) 12.0 Hz), 4.34 (t, 1H, J ) 4.5
Hz), 4.10 (t, 1H, J ) 6.9 Hz), 3.93 (dd, 1H, J ) 9.0, 6.9 Hz),
3.75 (t, 1H, J ) 8.1 Hz), 3.68 (t, 1H, J ) 10.2 Hz), 3.43 (m,
3H), 2.92 (s, 1H), 2.14 (m, 1H), 1.44 (s, 3H), 1.35 (s, 3H); 13C
NMR δ 138.17, 137.82, 128.32, 128.30, 127.97, 127.65, 127.62,
127.47, 108.89, 82.06, 79.92, 75.33, 74.06, 73.33, 73.30, 70.63,
69.94, 41.12, 28.12, 26.03. Anal. (C24H30O6) C, H.
1L-1,2,3-O-Ben zyl-6-d eoxy-6-(ben zyloxym eth yl)-ch ir o-
in ositol (12a ) a n d 1D-4,5,6-O-Ben zyl-3-d eoxy-3-(ben zyl-
oxym eth yl)-m yo-in ositol (12b). To a suspension of NaH (81
mg, 2.01 mmol) in DMF (2 mL) was added a solution of
compound 11a (278 mg, 0.67 mmol) in DMF (3 mL) at 0 °C.
After 30 min, BnBr (0.24 mL, 2.01 mmol) was added dropwise
and the solution was stirred for 5 h. The mixture was poured
into ice water and extracted with EtOAc (3 × 50 mL). The
organic phase was washed with H2O (50 mL) and brine (50
mL) and dried over MgSO4. The solvent was evaporated and
the residue was used directly in the next step.
1L-2-Allyl-1,3,4,5-O-Ben zyl-6-deoxy-6-(ben zyloxym eth yl)-
ch ir o-in osit ol (13a ) a n d 1D-1-Allyl-2,4,5,6-O-Ben zyl-3-
d eoxy-3-(ben zyloxym eth yl)-m yo-in ositol (13b). To a sus-
pension of NaH (32 mg, 0.78 mmol) in DMF (5 mL) was added
dropwise the above product (310 mg, 0.52 mmol) in DMF (5
mL) at 0 °C. After 30 min, BnBr (93 µL, 0.78 mmol) and n-Bu4-
NI (20 mg) were added, and the solution was stirred overnight.
The mixture was poured into ice water (20 mL) and extracted
with Et2O (3 × 30 mL). The organic phase was washed with
H2O (30 mL) and brine (30 mL) and dried over MgSO4. The
crude product was purified by column chromatography on
silica gel (EtOAc/hexane 1:4) to give the product 13a (356 mg,
99%): 1H NMR δ 7.30 (m, 25H), 5.94 (m, 1H), 5.26 (dd, 1H, J
) 17.4, 1.5 Hz), 5.14 (dd, 1H, J ) 10.2, 1.2 Hz), 4.89, 4.79 (ABq,
2H, J ) 10.5 Hz), 4.86, 4.82 (ABq, 2H, J ) 10.8 Hz), 4.74,
4.54 (ABq, 2H, J ) 12.0 Hz), 4.59, 4.54 (ABq, 2H, J ) 11.4
Hz), 4.50, 4.40 (ABq, 2H, J ) 12.0 Hz), 4.08 (m, 3H), 4.00-
3.89 (br m, 2H), 3.71 (dd, 1H, J ) 8.4, 4.5 Hz), 3.62-3.56 (m,
2H), 3.38 (t, 1H, J ) 9.9 Hz), 2.65 (m, 1H); 13C NMR δ 138.96,
138.87, 138.53, 138.17, 135.13, 128.37, 128.29, 128.26, 128.19,
128.16, 127.90, 127.82, 127.58, 127.55, 127.48, 127.41, 127.35,
83.09, 81.97, 79.64, 78.42, 75.94, 75.70, 74.89, 73.06, 72.78,
72.51, 71.64, 66.62, 40.35. Compound 13b was prepared in the
same manner in 99% yield: 1H NMR δ 7.35-7.17 (m, 25H),
5.96 (m, 1H), 5.34 (dd, 1H, J ) 17.4, 1.5 Hz), 5.17 (dd, 1H, J
) 10.8, 0.9 Hz), 4.98, 4.82 (ABq, 2H, J ) 11.1 Hz), 4.94, 4.82
(ABq, 2H, J ) 10.5 Hz), 4.84. 4.72 (ABq, 2H, J ) 10.5 Hz),
4.51, 4.43 (ABq, 2H, J ) 13.8 Hz), 4.40 (s, 2H), 4.20 (t, 2H, J
) 5.4 Hz), 4.16 (m, 1H), 4.00 (t, 1H, J ) 9.0 Hz), 3.69-3.44
(br m, 4H). 3.36 (dd, 1H, J ) 9.9, 2.1 Hz), 1.87 (m, 1H).
1L-1,3,4,5-O-Ben zyl-6-deoxy-6-(ben zyloxym eth yl)-ch ir o-
in ositol (14a ) a n d 1D-2,4,5,6-O-Ben zyl-3-d eoxy-3-(ben zyl-
oxym eth yl)-m yo-in ositol (14b). A solution of compound 13a
(356 mg, 0.52 mmol) in EtOH (10 mL) was refluxed with RhCl-
(PPh3)3 (25 mg, 0.026 mmol) and DABCO (147 mg, 1.3 mmol)
for 5 h. After evaporation, Et2O (100 mL) was added, and the
organic phase was washed with 3 N HCl (20 mL), H2O (20
mL) and brine (20 mL) and dried over MgSO4. After concen-
tration, the residue was dissolved in acetone/1 N HCl (20 mL,
9:1) and the solution was refluxed for 4 h. After evaporation,
Et2O (100 mL) was added and the solution was washed with
aqueous NaHCO3 (20 mL), H2O (20 mL) and brine (20 mL)
and dried over MgSO4. The crude product was purified by
column chromatography on silica gel (EtOAc/hexane 1:4) to
give the product 14a (294 mg, 88%): [R]D -14.6° (c 1.1, CHCl3);
1H NMR δ 7.31 (m, 25H), 4.84, 4.80 (ABq, 2H, J ) 11.1 Hz),
4.83, 4.74 (ABq, 2H, J ) 10.5 Hz), 4.60, 4.54 (ABq, 2H, J )
11.7 Hz), 4.51, 4.49 (ABq, 2H, J ) 8.4 Hz), 4.50, 4.48 (ABq,
2H, J ) 8.4 Hz), 4.02 (t, 1H, J ) 3.0 Hz), 4.01-3.87 (m, 2H).
3.75-3.60 (br m, 3H), 3.46 (t, 1H, J ) 9.3 Hz), 2.65 (m, 1H),
2.40 (d, 1H, J ) 7.8 Hz); 13C NMR δ 138.75, 138.68, 138.41,
138.21, 138.10, 128.38, 128.36, 128.32, 127.99, 127.95, 127.89,
127.68, 127.61, 127.53, 127.44, 82.87, 82.21, 78.10, 75.41,
73.19, 72.80, 72.18, 71.74, 66.48, 39.50. Anal. (C42H44O6) C,
H. Compound 14b was prepared in the same manner in 100%
yield: [R]D +20.2° (c 4.6, CHCl3); 1H NMR δ 7.27 (m, 25H),
4.93, 4.82 (ABq, 2H, J ) 10.8 Hz), 4.89 (s, 2H), 4.82, 4.74 (ABq,
To a solution of the above crude product in MeOH (20 mL)
was added concentrated HCl (0.10 mL) at room temperature
and the solution was stirred for 24 h. After evaporation, the
residue was purified by column chromatography on silica gel
(EtOAc/hexane 1:1) to give the product (333 mg, 90%): 1H NMR
δ 7.30 (m, 20H), 4.88, 4.63 (ABq, 2H, J ) 11.4 Hz), 4.83, 4.69
(ABq, 2H, J ) 11.1 Hz), 4.59 (s, 2H), 4.51, 4.48 (ABq, 2H, J )
12.3 Hz), 4.18 (s, 1H), 4.02 (dd, 1H, J ) 7.8, 6.0 Hz), 3.88 (m,
1H), 3.79-3.58 (br m, 4H), 2.57 (m, 3H); 13C NMR δ 138.50,
138.42, 138.11, 128.55, 128.37, 128.34, 128.27, 127.94, 127.84,
127.81, 127.68, 127.61, 127.57, 127.51, 81.22, 82.80, 78.36,
74.78, 74.69, 73.19, 72.60, 71.50, 69.75, 67.11, 41.21. Com-
pound 12b was prepared in the same manner in 98% yield:
1
mp 103-104 °C; [R]D +10.8° (c 3.2, CHCl3); H NMR δ 7.30
(m, 20H), 4.94, 4.91 (ABq, 2H, J ) 11.1 Hz), 4.92, 4.50 (ABq,
2H, J ) 11.1 Hz), 4.89, 4.79 (ABq, 2H, J ) 11.1 Hz), 4.49,
4.48 (ABq, 2H, J ) 12.6 Hz), 4.25 (m, 1H), 4.34-3.81 (m, 3
H), 3.73 (dd, 1H, J ) 9.0, 3.0 Hz), 3.58-3.49 (m, 2H), 3.33 (s,
1H), 2.41 (d, 1H, J ) 5.1 Hz), 1.75 (m, 1H); 13C NMR δ 138.58,
138.29, 137.50, 137.49, 128.49, 128.37, 127.96, 127.88, 127.71,
127.63, 127.51, 86.50, 82.26, 77.30, 75.61, 75.48, 75.40, 74.50,
73.47, 70.79, 68.71, 43.35. Anal. (C35H38O6) C, H.
1L-2-Allyl-3,4,5-O-Ben zyl-6-d eoxy-6-(ben zyloxym eth yl)-
ch ir o-in ositol a n d 1D-1-Allyl-4,5,6-O-Ben zyl-3-d eoxy-3-
(ben zyloxym eth yl)-m yo-in ositol. A solution of compound
12a (333 mg, 0.60 mmol) in toluene (10 mL) was refluxed with
Bu2SnO (179 mg, 0.72 mmol) until a clear solution was
obtained (approximately 2 h). The solution was evaporated,
and the residue was dried in vacuo and taken up in 5 mL of
dry DMF. To this solution were added CsF (300 mg, 1.9 mmol)
and allyl bromide (90 µL, 1.04 mmol) at -50 °C. The mixture
was warmed to room temperature overnight, then poured into
ice water and extracted with EtOAc (3 × 50 mL). The organic
phase was washed with H2O (3 × 50 mL) and brine (50 mL)
and dried over MgSO4. The crude product was purified by
column chromatography on silica gel (EtOAc/hexane 1:3) to
give the product (310 mg, 87%): [R]D -21.7° (c 1.14, CHCl3);

3050 J ournal of Medicinal Chemistry, 2000, Vol. 43, No. 16
Hu et al.
2H, J ) 11.1 Hz), 4.53, 4.45 (ABq, 2H, J ) 11.4 Hz), 4.42,
4.40 (ABq, 2H, J ) 12.0 Hz), 4.13 (m, 1H), 3.83 (t, 1H, J ) 9.3
Hz), 3.70 (dd, 1H, J ) 8.7, 4.2 Hz), 3.64 (m, 1H), 3.60-3.52
(m, 2H), 3.48 (t, 1H, J ) 9.0 Hz), 2.18 (d, 1H, J ) 4.2 Hz),
1.96 (m, 1H); 13C NMR δ 139.06, 138.46, 138.13, 138.08, 128.53,
128.38, 128.34, 128.33, 128.19, 128.01, 127.94, 127.82, 127.76,
127.68, 127.59, 127.55, 127.37, 86.69, 82.10, 78.72, 75.80,
75.53, 75.49, 75.16, 75.15, 73.07, 67.08, 44.13. Anal. (C42H44O6)
C, H.
1L-1,3,4,5-O-Ben zyl-6-deoxy-6-(ben zyloxym eth yl)-ch ir o-
in ositol 2-[Ben zyl (R)-2-O-m eth yl-3-O-octa d ecyl p h os-
p h a te] (15a ) a n d 1D-2,4,5,6-O-Ben zyl-3-d eoxy-3-(ben zyl-
oxym et h yl)-m yo-in osit ol 2-[Ben zyl (R)-2-O-m et h yl-3-O-
octa d ecyl p h osp h a te] (15b). To a suspension of diisopropyl-
ammonium tetrazolide (80 mg, 0.46 mmol) in dry CH2Cl2 (2
mL) was added dropwise under N2 at room temperature
O-benzyl-N,N,N′,N′-tetraisopropylphosphorodiamidite (0.23 mL,
0.64 mmol), followed by a solution of compound 14a (274 mg,
0.42 mmol) in dry CH2Cl2 (3 mL). The mixture was stirred
overnight at room temperature, then aqueous NaHCO3 solu-
tion (10 mL) was added. The aqueous phase was extracted with
CH2Cl2 (3 × 10 mL). The combined organic phases were dried
over MgSO4 and evaporated. The residue was purified by
column chromatography on silica gel previously deactivated
with Et3N (1 mL) (EtOAc/hexane 1:4) to give the phosphora-
midite intermediate which was used directly in the next step.
CHCl3/MeOH 1:1); 1H NMR (CDCl3/CD3OD 1:1, TMS) δ 4.34-
4.07 (m, 4H), 4.01 (dd, 1H, J ) 9.3, 6.0 Hz), 3.87 (dd, 1H, J )
11.7, 5.1 Hz), 3.80 (t, 1H, J ) 8.7 Hz), 3.64-3.45 (m, 10H),
2.37 (m, 1H), 1.58 (m, 2H), 1.27 (m, 30H), 0.89 (t, 3H, J ) 6.0
Hz); 13C NMR (CDCl3/CD3OD 1:1) δ 79.95 (J ) 6.0 Hz), 79.53
(J ) 7.6 Hz), 74.87, 72.64 (J ) 5.5 Hz), 72.43, 70.52, 69.91,
69.57, 66.73 (J ) 5.5 Hz), 59.21, 58.19, 47.31, 32.48, 30.94,
30.23, 30.19, 30.09, 30.05, 29.90, 26.58, 23.20, 14.35; 31P NMR
δ -0.09. Anal. (C29H59O11P‚0.3H2O) C, H. Compound 3 was
prepared in the same manner in 88% yield: mp 60-61 °C;
[R]D -4.9° (c 0.42, CHCl3/MeOH 1:1); 1H NMR (CDCl3/CD3-
OD 2:1, TMS) δ 4.29 (m, 1H), 4.17-3.82 (m, 5H), 3.73 (t, 1H,
J ) 9.6 Hz), 3.60-3.44 (m, 8H), 3.35 (m, 1H), 3.39 (t, 1H, J )
8.7 Hz),1.58 (m, 3H), 1.27 (m, 30H), 0.89 (t, 3H, J ) 6.3 Hz);
13C NMR (CDCl3/CD3OD 2:1) δ 81.33, 79.41, 78.05, 72.32,
71.89, 69.97, 69.75, 69.44, 66.49, 61.75, 58.09, 44.73, 32.30,
30.88, 30.06, 30.01, 29.91, 29.87, 29.73, 26.40, 23.04, 14.24;
31P NMR (DMSO-d6) δ 0.02. Anal. (C29H59O11P‚2H2O) C, H.
1L-1,3,4,5-O-Ben zyl-6-deoxy-6-(ben zyloxym eth yl)-ch ir o-
in ositol 2-[Ben zyl (R)-2-O-m eth yl-3-O-octa d ecylca r bon -
a te] (17a ) a n d 1D-2,4,5,6-O-Ben zyl-3-d eoxy-3-(ben zyloxy-
m eth yl)-m yo-in ositol 2-[Ben zyl (R)-2-O-m eth yl-3-O-octa -
d ecylca r bon a te] (17b). To a solution of compound 14a (60
mg, 94 µmol) in toluene (2 mL) was added 1,1′-carbonyldiimi-
dazole (24 mg, 0.14 mmol), and the mixture was refluxed for
40 min. The mixture was cooled to room temperature and
diluted with EtOAc (30 mL). The solution was washed with
H2O (2 × 10 mL) and brine (10 mL), dried over MgSO4, and
evaporated. The crude product was used directly in the next
step.
To a solution of 2-O-methyl-1-O-octadecyl-sn-glycerol (150
mg, 0.42 mmol) and 1H-tetrazole (50 mg, 0.7 mmol) in dry CH2-
Cl2 (2 mL) was added a solution of the preceding phosphora-
midite in dry CH2Cl2 (3 mL) under N2 at room temperature.
The mixture was stirred overnight at room temperature, then
aqueous NaHCO3 solution (10 mL) was added. The aqueous
phase was extracted with CH2Cl2 (3 × 10 mL). The combined
organic phases were dried over MgSO4 and evaporated. The
residue was used directly in the next step.
To a solution of the above crude product in toluene (2 mL)
was added DBU (17 µL, 0.010 mmol) and 2-O-methyl-1-O-
octadecyl-sn-glycerol (35 mg, 0.10 mmol). The mixture was
refluxed for 4 h, then evaporated and the product was isolated
by column chromatography on silica gel (EtOAc/hexane 1:10)
to yield the product (52 mg, 54%): [R]D -15.8° (c 2.32, CHCl3);
1H NMR δ 7.10 (m, 25 H), 5.11 (dd, 1H, J ) 10.2, 2.7 Hz),
4.84, 4.76 (ABq, 2H, J ) 10.5 Hz), 4.82, 4.76 (ABq, 2H, J )
10.8 Hz), 4.60, 4.50 (ABq, 2H, J ) 11.7 Hz), 4.51 (s, 2H), 4.54,
4.48 (ABq, 2H, J ) 11.7 Hz), 4.27 (dd, 1H, J ) 11.1, 3.9 Hz),
4.18 (d, 1H, J ) 5.7 Hz), 4.13 (m, 1H), 4.04-3.96 (br m, 2H),
3.78 (t, 1H, J ) 9.0 Hz), 3.67 (dd, 1H, J ) 9.9, 4.2 Hz), 3.56-
3.38 (br m, 9 H), 2.59 (m, 1H), 1.57 (m, 2H), 1.25 (m, 30H),
0.88 (t, 3H, J ) 6.3 Hz); 13C NMR δ 154.81, 138.72, 138.56,
138.40, 138.33, 138.09, 128.36, 128.31, 128.29, 128.25, 127.98,
127.84, 127.73, 127.59, 127.54, 127.51, 127.42, 82.85, 80.01,
78.16, 78.01, 77.93, 76.07, 75.65, 75.55, 73.31, 72.84, 72.67,
71.85, 69.71, 67.29, 66.42, 58.10, 40.68, 31.90, 29.68, 29.59.
29.48, 29.34, 29.20, 26.05, 22.68, 14.12. Compound 17b was
prepared in the same manner in 89% yield: [R]D +12.3° (c 1.45,
CHCl3); 1H NMR δ 7.28 (m, 25 H), 4.92-4.75 (br m, 7H), 4.46-
4.35 (br m, 4H), 4.28 (dd, 1H, J ) 11.4, 3.9 Hz), 4.22 (m, 1H),
4.19 (dd, 1H, J ) 11.4, 5.7 Hz), 4.06 (t, 1H, J ) 9.0 Hz), 3.69-
3.37 (m, 12H), 2.01 (m, 1H), 1.54 (m, 2H), 1.25 (bm, 30H), 0.88
(t, 3H, J ) 6.3 Hz); 13C NMR δ 154.80, 138.59, 138.44, 138.10,
138.06, 128.36, 128.27, 128.17, 128.01, 127.81, 127.76, 127.71,
127.63, 127.49, 127.45, 86.34, 80.90, 80.08, 78.06, 77.98, 76.58,
75.81, 75.65, 75.22, 74.44, 73.08, 71.85, 69.59, 67.41, 66.82,
58.06, 43.72, 31.90, 29.68, 29.58, 29.47, 29.33, 26.03, 22.67,
24.11. Anal. (C65H88O10) C, H.
To a solution of the preceding phosphite in dry CH2Cl2 (10
mL) was added t-BuOOH (0.15 mL, 0.64 mmol) at 0 °C. The
mixture was warmed to room temperature and stirred for 4
h. The solvent was evaporated, and the product was isolated
by column chromatography on silica gel (EtOAc/hexane 1:3)
to yield 15a (270 mg, 55%): [R]D -4.4° (c 0.82, CHCl3); 1H NMR
δ 7.25 (m, 30H), 5.05-4.42 (m, 12H), 4.25 (m, 1H), 4.08-3.98
(m, 4H), 3.79 (t, 1H, J ) 9.0 Hz), 3.59 (m, 2H), 3.40-3.27 (br
m, 8H), 2.52 (m, 1H), 1.48 (m, 2H), 1.26 (m, 30H), 0.88 (t, 3H,
J ) 6.0 Hz); 13C NMR δ 138.72, 138.61, 138.51, 138.45, 138.38,
138.08, 136.05, 136.03, 128.37, 128.32, 128.27, 128.22, 128.19,
128.14, 127.95, 127.80, 127.65, 127.57, 127.41, 82.45, 80.29,
78.43 (J ) 7.6 Hz), 77.98, 77.42, 75.49, 73.20, 72.89, 72.77,
71.69, 69.47, 68.95 (J ) 5.4 Hz), 67.89, 66.52, 66.48, 57.87,
40.39, 31.87, 29.65, 29.61, 29.56, 29.51, 29.45, 29.31, 25.98,
22.64, 14.08; 31P NMR δ -0.95, -1.04. Anal. (C71H95O11P) C,
H. Compound 15b was prepared in the same manner in 38%
1
yield: [R]D +21.0° (c 0.80, CHCl3); H NMR δ 7.28 (m, 30H),
5.06-4.78 (br m, 8H), 4.50 (d, 1H, J ) 10.8 Hz), 4.45-4.33
(m, 5H), 4.13-3.97 (br m, 3H), 3.66-3.52 (br m, 3H), 3.43-
3.28 (br m, 9H), 1.92 (m, 1H), 1.48 (m, 2H), 1.26 (m, 30H),
0.88 (t, 3H, J ) 6.3 Hz); 13C NMR δ 138.90, 138.53, 138.49,
138.37, 138.10, 138.08, 128.48, 128.43, 128.35, 128.19, 128.15,
128.12, 128.03, 127.81, 127.74, 127.71, 127.65, 127.63, 127.56,
127.36, 86.27, 81.70, 80.42, 78.67 (J ) 7.6 Hz), 77.96, 75.83,
75.50, 75.39, 75.32, 75.20, 73.05, 71.79, 69.36 (J ) 6.0 Hz),
69.07, 66.79, 66.57, 57.93, 43.54, 30.89, 29.68, 29.63, 29.59,
29.54, 29.47, 29.33, 26.01, 25.74, 22.66, 14.11; 31P NMR δ
-0.74, -0.92. Anal. (C71H95O11P) C, H.
1L-6-Hyd r oxym eth yl-ch ir o-in ositol 2-[(R)-2-O-Meth yl-
3-O-oct a d ecylca r b on a t e] (5) a n d 1D-3-Hyd r oxym et h yl-
m yo-in ositol 1-[(R)-2-O-Meth yl-3-O-octa d ecylca r bon a te]
(6). A solution of compound 17a (60 mg, 0.058 mmol) in
t-BuOH (5 mL) was hydrogenated for 6 h in a Parr shaker
under 60 psi of H2 at room temperature, using 20% Pd(OH)2/C
(30 mg) as catalyst. The catalyst was filtered off through a
Celite pad, and the filtrate was evaporated. The residue was
dried in vacuo to give white powder (28 mg, 83%): mp 62 °C
1L-6-Hyd r oxym eth yl-ch ir o-in ositol 2-[(R)-2-O-Meth yl-
3-O-octa d ecyl h yd r ogen p h osp h a te] (2) a n d 1D-3-Hy-
d r oxym eth yl-m yo-in ositol 1-[(R)-2-O-Meth yl-3-O-octa d e-
cyl h yd r ogen p h osp h a te] (3). A solution of compound 15a
(280 mg, 0.242 mmol) in t-BuOH (10 mL) was hydrogenated
1
for 5 h in
a
Parr shaker under 60 psi of H2 at room
dec; [R]D -25.8° (c 0.96, CHCl3/MeOH 1:1); H NMR (CDCl3/
temperature, using 20% Pd(OH)2/C (103 mg) as catalyst. The
catalyst was filtered off through a Celite pad, and the filtrate
was evaporated. The residue was dried in vacuo to give a white
powder (131 mg, 88%): mp 64 °C dec; [R]D -18.8° (c 1.92,
CD3OD 1:1, TMS) δ 4.37 (m, 1H), 4.32 (dd, 1H, J ) 11.7, 3.9
Hz), 4.25 (t, 1H, J ) 3.0 Hz), 4.19(dd, 1H, J ) 11.4, 5.7 Hz),
4.01 (dd, 1H, J ) 9.3, 5.7 Hz), 3.91 (dt, 1H, J ) 11.1, 5.4 Hz),
3.82 (d, 1H, J ) 9.6 Hz), 3.64-3.34 (m, 10H), 2.38 (m, 1H),

Phosphatidylinositols and Carbonate Surrogates
J ournal of Medicinal Chemistry, 2000, Vol. 43, No. 16 3051
1.58 (m, 2H), 1.27 (m, 30H), 0.89 (t, 3H, J ) 6.3 Hz); 13C NMR
(CDCl3/CD3OD 1:1) δ 155.65, 79.44, 78.67, 75.00, 72.40, 71.73,
70.68, 69.95, 68.33, 67.66, 59.47, 58.13, 47.59, 32.47, 30.90,
30.21, 30.07, 30.00, 29.89, 26.58, 23.19, 14.28. Anal. (C30H58O10‚
0.5H2O) C, H. Compound 6 was prepared in the same manner
in 72% yield: [R]D -15.8° (c 0.74, CHCl3/MeOH 1:1); 1H NMR
(CDCl3/CD3OD 2:1, TMS) δ 4.35-4.29 (m, 2H), 4.20 (dd, 1H,
J ) 11.4, 5.7 Hz), 3.97-3.85 (m, 3H), 3.74 (t, 1H, J ) 9.6 Hz),
3.63 (m, 1H), 3.55-3.29 (m, 9H), 1.58 (m, 3H), 1.27 (m, 30H),
0.89 (t, 3H, J ) 6.3 Hz); 13C NMR (CDCl3/CD3OD 2:1) δ 155.36,
80.96, 78.41, 78.12, 72.31, 70.82, 69.86, 69.29, 68.58, 67.54,
61.62, 58.11, 44.78, 32.39, 30.68, 30.03, 29.88, 29.82, 29.71,
26.39, 23.01, 14.24. Anal. (C30H58O10‚0.5H2O) C, H.
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Compound 4 was prepared from 1D-2,4,5,6-tetra-O-benzyl-
5e
3-deoxy-myo-inositol
in the same manner: [R]D -28.3° (c
1.30, CHCl3/MeOH 1:1); 1H NMR (CDCl3/CD3OD 1:1, TMS) δ
4.49 (dd, 1H, J ) 9.9, 3.0 Hz), 4.32 (dd, 1H, J ) 11.4, 3.6 Hz),
4.21 (m, 2H), 3.83 (t, 2H, J ) 9.3 Hz), 3.63 (m, 1H), 3.55 (m,
2H), 3.49 (m, 5H), 3.26 (t, 1H, J ) 9.0 Hz), 2.13 (dt, 1H, J )
4.5, 14.1 Hz), 1.56 (m, 3H), 1.40-1.25 (m, 30H), 0.89 (t, 3H, J
) 6.3 Hz); 13C NMR (CDCl3/CD3OD 1:1) δ 155.49, 81.32, 78.55,
72.36, 71.04, 69.84, 68.46, 67.60, 66.39, 58.11, 35.84, 32.37,
30.12, 30.05, 29.98, 29.91, 29.80, 26.49, 23.10, 14.25. Anal.
(C29H56O9) C, H.
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A.; Nahorski, S. R. 1D-myo-Inositol 1,4,5-trisphosphate and 1D-
myo-inositol 1,3,4,5-tetrakisphosphate analogues modified at
C-3; synthesis of 1D-3-C-(trifluoromethyl)-myo-inositol 1,4,5-
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dideoxy, and 1D-2,3,6-trideoxy-myo-inositol 1,4,5-trisphosphate
from quebrachitol, their binding affinities, and calcium release
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ki, A. P.; Kiddle, J . J .; Frew, T.; Berggren, M.; Powis, G.
Ack n ow led gm en t. This work was supported by the
National Institutes of Health (Grant No. CA61015), by
the Department of Defense (DAMD 17-93-V-3018), and
in part by the Office of Naval Research.
Su p p or tin g In for m a tion Ava ila ble: X-ray structural
data for 10 is available free of charge via the Internet at http://
pubs.acs.org.
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