Conformationally Constrained Analogues of Diacylglycerol. 20. The Search for an Elusive Binding Site on Protein Kinase C through Relocation of the Carbonyl Pharmacophore Along the sn-1 Side Chain of 1,2-Diacylglycerol Lactones

Article abstract of DOI:10.1021/jm030454h

Previous studies with 1,2-diacylglycerol (DAG) lactones, which behave as high-affinity ligands for protein kinase C (PK-C), have established the importance of maintaining intact the pharmacophore triad of two carbonyl moieties (sn-1 and sn-2) and the prim

Full text of DOI:10.1021/jm030454h

644
J . Med. Chem. 2004, 47, 644-655
Con for m a tion a lly Con str a in ed An a logu es of Dia cylglycer ol. 20. Th e Sea r ch for
a n Elu sive Bin d in g Site on P r otein Kin a se C th r ou gh Reloca tion of th e
Ca r bon yl P h a r m a cop h or e Alon g th e sn -1 Sid e Ch a in of 1,2-Dia cylglycer ol
La cton es
Hirokazu Tamamura,†,‡ Dina M. Sigano,† Nancy E. Lewin,§ Peter M. Blumberg,§ and Victor E. Marquez*,†
Laboratory of Medicinal Chemistry, Center for Cancer Research, National Cancer Institute at Frederick,
National Institutes of Health, Frederick, Maryland 21702, and Laboratory of Cellular Carcinogenesis & Tumor Promotion,
Center for Cancer Research, National Cancer Institute, National Institutes of Health, Bethesda, Maryland 20892
Received September 12, 2003
Previous studies with 1,2-diacylglycerol (DAG) lactones, which behave as high-affinity ligands
for protein kinase C (PK-C), have established the importance of maintaining intact the
pharmacophore triad of two carbonyl moieties (sn-1 and sn-2) and the primary alcohol. In
addition, docking studies of DAG-lactones into an empty C1b receptor of PK-Cδ (as it appears
in complex with phorbol 13-O-acetate) have revealed that in either of the two possible binding
alternatives (sn-1 or sn-2) only one carbonyl group of the DAG-lactone is involved in binding.
Therefore, the unknown receptor for the orphaned carbonyl appears to lie outside the boundaries
of this binary complex, possibly residing at the membrane or near the membrane-protein
interface. A strategy to locate the optimal location of the unengaged carbonyl was conceived
by utilizing a small group of DAG-lactones (1-4) with a highly branched chain adjacent to the
sn-2 carbonyl such that sn-2 binding is favored. With these compounds, various locations of
the sn-1 carbonyl along the side chain were tested for their binding affinity for PK-C. The
results indicate that the location of the side chain sn-1 carbonyl in a DAG-lactone must have
perfect mimicry to the sn-1 carbonyl of the parent DAG for it to display high binding affinity.
A proposed model from this work is that the missing pharmacophore in the ternary complex,
which includes the membrane, is close to the membrane-protein interface.
In tr od u ction a n d Ba ck gr ou n d
lipids.7,8 This association is strongly facilitated by DAG,
which is generated as a result of a stimulus-initiated
activation of phospholipase C.9 The accepted dogma has
been that these isozymes are cytosolic in the inactive
state and, as part of the activation process, translocate
to the inner leaflet of the cellular membrane.10,11 The
binding of PK-C to the plasma membrane is transient
and regulated by the association of its C1 domain with
DAG in the membrane.12
Protein kinase C (PK-C) is a family of serine/threo-
nine-specific isozymes that are regulated by phospho-
rylation, by calcium, and by association with phospho-
lipids and sn-1,2-diacylglycerol (DAG).1-3 The PK-C
family comprises 10 isozymes grouped into three
classes: conventional (R, â, γ), novel (δ, ꢀ, η, θ), and
atypical (ú, ι/λ). In addition, PK-C µ and ν are considered
by some to constitute a fourth class and by others to
comprise a distinct family called protein kinase D.4 All
members contain a C-terminal kinase domain with
serine/threonine specific kinase activity and an N-
terminal regulatory domain. The regulatory domain
contains two key functionalities: an autoinhibitory
sequence and one or two membrane-targeting domains
(C1 and C2).
The central role of PK-C in cell signal transduction
has been well established over the past 2 decades since
its discovery5 with the lipophilic second messenger,
DAG, playing a prominent role in this cellular signal
transduction.6 Both the classical (R, â, and γ) and novel
(δ, ꢀ, η, θ) PK-C isozymes are thought to become
activated as a result of association of the cytosolic
enzyme with membranes containing acid phospho-
The classical and novel PK-C isozymes each contain
two small (∼50 residues) zinc-finger-like domains (C1a
and C1b) consisting of two â-sheets and a small R-helix.
The crystal structure of an isolated C1 domain from PK-
Cδ in complex with phorbol 13-O-acetate13 showed that
when the ligand is bound in the pocket between the
â-sheets, a continuous hydrophobic surface is created
over the top third of the domain, which in turn promotes
its insertion into the membrane. This binary complex
structure confirmed the importance of hydrogen bonding
of the main phorbol ester pharmacophores, C3 (CdO),
C4 (OH), and C20 (OH), in agreement with previous
SAR studies showing that any modification to these key
functional groups abrogated or reduced the activity of
the phorbol esters (Figure 1).14,15 The structure also
revealed that the C12 hydroxyl and C13 acetate moieties
were not hydrogen-bonded to any residue on the C1
domain. However, SAR studies have also shown that
lipophilic acyl chains at these positions are important
for activity because they bind to membrane lipids in the
ternary complex.16,17 The C9 (OH), although originally
* To whom correspondence should be addressed. Phone: 301-846-
5954. Fax: 301-846-6033. E-mail: marquezv@dc37a.nci.nih.gov.
† Laboratory of Medicinal Chemistry.
‡ Present address: Graduate School of Pharmaceutical Sciences,
Kyoto University, Sakyo-ku, Kyoto 606-8501, J apan.
§ Laboratory of Cellular Carcinogenesis & Tumor Promotion.
10.1021/jm030454h This article not subject to U.S. Copyright. Published 2004 by the American Chemical Society
Published on Web 12/31/2003

Analogues of Diacylglycerol
J ournal of Medicinal Chemistry, 2004, Vol. 47, No. 3 645
F igu r e 1. Pharmacophores (yellow) of phorbol, PMA, DAG, and DAG-lactone.
believed to be involved in binding, appeared intra-
molecularly hydrogen-bonded to the C13 carbonyl ester.
While it has been suggested that such an intramolecular
arrangement prevents these polar groups from residing
in the lipid phase with unsatisfied hydrogen bonds,13 it
is also possible that this motif could interact at the
protein-lipid interface as an additional pharmacophore
not revealed in the binary complex. Corresponding to
the phorbol ester pharmacophores are C1 (CdO), C2
(CdO), and C3 (CH2OH) in DAG, although a substantial
difference in binding affinity (g3 orders of magnitude)
is found favoring the phorbol esters (Figure 1).18,19 On
the basis of the spatial correspondence between the key
F igu r e 2. Hydrogen-bonding interaction of DAG-lactone at
the C1 domain of PK-Cδ in the sn-2 binding mode.
oxygen atoms in phorbol and DAG, a pharmacophore-
guided approach was used to design highly potent PK-C
ligands based on the DAG structure where the glycerol
backbone is constrained into a five-member lactone ring
(DAG-lactone, Figure 1).20-22
absolutely essential;25 thus, it is probable that this key
pharmacophore interacts with phospholipid headgroups,
other structural elements outside the C1 domain to
which the DAG-lactone is bound, and even amino acid
residues on the adjacent, unoccupied C1 domain. There-
fore, in the absence of a solved ternary complex struc-
ture, which involves the DAG-lactone, the C1 domain(s)
and the membrane, the optimal location of this orphan
sn-1 pharmacophore cannot be determined with confi-
dence. A small number of DAG-lactones, which are
described in the present work, were designed and
synthesized to find the optimal location of this elusive
pharmacophore.
Modeling studies using the crystal coordinates of the
C1b domain of PK-Cδ in complex with phorbol 13-O-
acetate revealed two possible binding modes (sn-1 and
sn-2) when a DAG-lactone was docked into the empty
C1b domain.20 Although the carbonyl moieties are not
equivalent, they display a similar hydrogen-bonding
network in two distinct orientations with participation
from the primary OH and only one of the carbonyl
groups, to Thr242, Leu251, and Gly253, akin to that
seen with phorbol 13-O-acetate. The sn-1 binding mode
in either DAG or DAG-lactones is defined as that in
which only the sn-1 carbonyl is directly bound to the
protein, while the sn-2 binding mode is defined as that
in which only the sn-2 carbonyl is directly bound to the
protein. Since the DAG-lactones compete with phorbol
esters for the same binding site,23 a common binding
mode between these two classes of ligands would require
at least one CdO group of the DAG-lactone to be involved
in binding to the C1 domain of the enzyme.
While we observe that in terms of hydrogen bonding
both the sn-1 and sn-2 binding modes are allowed, the
disposition and nature of the acyl chain, particularly if
it is highly branched, can have a direct influence on the
binding orientation of the DAGs.24 Indeed, we found that
in general DAG-lactones prefer to bind sn-2, and this
sn-2 binding predilection can be enhanced by placing
larger bulky groups R to the lactone carbonyl.21 It is in
this binding mode (Figure 2) that DAG-lactones exhibit
a large increase in binding affinity when compared to
their open-chain counterparts.21 While in this binding
mode the sn-1 carbonyl appears to be unengaged with
the C1 domain in the binary complex, its presence is
Design a n d Syn th esis
To better understand the more subtle details of the
interaction of the sn-1 carbonyl, we undertook the task
to explore the preferred location of this pharmacophore
to complement its unidentified receptor. By placement
of a large bulky group next to the sn-2 carbonyl and a
slimmer, linear chain in the sn-1 position, lactones (1-
4) were designed to favor sn-2 binding to the protein,
thus leaving the sn-1 carbonyl free to bind to the
unspecified receptor. In this series of compounds, the
sn-1 carbonyl is “moved” through the side chain in an
attempt to identify its optimal position for interacting
with its unidentified receptor.
Relative to the position of the sn-1 carbonyl in 1,
targets were selected with a one-carbon reduction (2),
achieved by reversing the ester functionality and thus
moving the carbonyl one position closer to the lactone
ring, and with a one-carbon elongation (3) and two-
carbon elongation (4), achieved by moving the carbonyl

646 J ournal of Medicinal Chemistry, 2004, Vol. 47, No. 3
Tamamura et al.
one and two positions, respectively, further away from
the lactone ring. It is noteworthy that since all the
As shown in Scheme 2, protection of the primary
hydroxyl group of 14 as its MPM ether (15), followed
by hydrolysis of the methyl glycoside and subsequent
oxidation, gave 17. R-Alkylation of lactone 17 followed
by dehydration gave 18 as a mixture of E and Z
R-alkylidene isomers, which were easily separated by
silica gel column chromatography. The MPM ether of
each isomer was then separately deprotected to give 19,
the common late-stage intermediate for the syntheses
of 2 and 3. For each isomer of 19 (E/Z), the primary
alcohol was subjected to Swern oxidation to the alde-
hyde (20) and further oxidation to the carboxylic acid
to give 21 (E/Z). DCC coupling with 1-nonanol and final
removal of the benzyl group gave the desired one-carbon
reduction reverse ester compounds 2E and 2Z. Alter-
natively, direct acylation of the primary alcohol of 19
(E/Z) followed by removal of the protecting group gave
3E and 3Z, the one-carbon elongated analogues.
To synthesize the longer two-carbon extension lactone
4, intermediate 14 underwent further transformations
to give the elongated chain (26, Scheme 3). The primary
alcohol of 26 was then protected as its MPM ether (27),
the methyl glycoside was hydrolyzed, and the resultant
secondary alcohol (28) was oxidized to give DAG-lactone
29. Successive R-alkylation followed by dehydration
gave 30 as a mixture of E and Z isomers, which were
separated by silica gel column chromatography. The
MPM ether of each isomer was then separately depro-
tected and acylated using octanoyl chloride. Final
removal of the benzyl group of 32E and 32Z gave,
respectively, 4E and 4Z as shown in Scheme 3.
compounds have the same empirical formula and iden-
tical calculated log P values (log P ) 6.61), they would
be expected to have similar hydrophobicities, at least
to a first approximation. However, since the compounds
can form different intramolecular H-bonding networks,
their movement into a lipid environment would involve
different desolvation energy penalties that could ad-
ditionally influence their binding affinity to some de-
gree.
A common intermediate, 14, was synthesized starting
from commercially available 3-benzyloxy-1,2-propanediol
(5), as outlined in Scheme 1. Protection of the primary
alcohol (6) followed by oxidation gave the requisite
ketone intermediate 7. Grignard reaction with allyl-
magnesium bromide followed by hydroboration and PCC
oxidation gave lactone 8. DIBAL reduction of the
carbonyl followed by conversion of the resulting lactol
(9) to the methyl glycoside (10) and silyl deprotection
gave 11. Oxidation of the primary alcohol to an aldehyde
(12), followed by reaction under Wittig conditions,
elongated the chain by one carbon (13). Final hydro-
boration and oxidation gave 14.
Biologica l Resu lts a n d Discu ssion
On the basis of our understanding that a DAG-lactone
like 1 with a large, branched R-alkylidene chain pref-
erentially binds in an sn-2 mode21 with the sn-1 carbonyl
devoid of a tangible contact with the C1 domain in the
binary complex, isomeric compounds 2-4 were designed
with the intent of finding the best location for this
pharmacophore in a phospholipid-containing ternary
complex. Compound 1 was identified in a previous study
as a very potent ligand with the characteristic activity
profile showing ca. 2-fold difference in binding affinity
favoring the Z isomer (Ki ) 2.3 versus 6.6 nM for the E
isomer).20 Because 1-4 are structurally similar and
have identical empirical formulas and calculated log P
values, their hydrophobicities should be similar; thus,
the only variable that remains is the displacement of
the sn-1 carbonyl along the fixed-length arm of a DAG-
lactone relative to the prototype compound 1. As already
mentioned, the displacement of the sn-1 carbonyl along
the chain is also accompanied by changes in the in-
tramolecular web of H-bonding, which could facilitate
or hinder the molecule’s movement into a lipid environ-
ment and influence the binding affinity. The transposi-
tion of the ester oxygen and carbonyl in compound 2,
which brings the position of the carbonyl closer to the
lactone ring, resulted in ca. 10-fold loss in binding
affinity for both the Z and E isomers (Table 1). Going
in the other direction, increasing the distance between
the lactone and the carbonyl of the side chain by one
(3) or two (4) carbons resulted in a 100- to 300-fold
decrease in binding affinity. The reproducible and
superior binding affinity in favor of the Z isomers in
Sch em e 1a
a
(a) TBDPSCl, imidazole, CH2Cl2, DMF; (b) PCC, 4 Å molecular
sieves, CH2Cl2; (c) allylmagnesium bromide, THF, 0 °C to room
temperature; (d) BH3‚SMe2, THF, -78 °C to room temperature;
(e) PCC, 4 Å molecular sieves, CH2Cl2; (f) DIBAL, PhMe, -78 °C;
(g) Dowex 50WX8-200, AcOH, MeOH, THF; (h) TBAF, THF, 0 °C
to room temperature; (i) DCC, DCAA, DMSO; then oxalic acid,
Et2O, -78 °C; (j) MePPh3Br, KOC(CH3)3, THF; (k) BH3‚SMe2,
THF, -78 to 0 °C, then NaBO3‚H2O, 0 °C to room temperature.

Analogues of Diacylglycerol
J ournal of Medicinal Chemistry, 2004, Vol. 47, No. 3 647
Sch em e 2a
a
(a) NaH, THF, 0 °C, then MPMCl, Bu4NI, ∆; (b) HCl, THF, H2O, 0 °C to room temperature; (c) TPAP, NMO, 4 Å molecular sieves,
CH2Cl2/CH3CN, 0 °C to room temperature; (d) LDA, THF, ((CH3)2CH)2CHCH2CHO, -78 °C; (e) MsCl, Et3N, 0 °C to room temperature;
(f) DBU, 0 °C to room temperature; (g) DDQ, H2O, CH2Cl2; (h) (COCl)2, DMSO, Et3N, CH2Cl2, -78 to 0 °C; (i) 2-methyl-2-butene, NaClO2,
NaH2PO4, (CH3)3COH, H2O; (j) n-C9H19OH, DMAP, DCC, pyridine, 0 °C to room temperature; (k) BCl3, CH2Cl2, -78 °C; (l) n-C8H17C(O)Cl,
pyridine, CH2Cl2, 0 °C.
Sch em e 3a
a
(a) DCC, DCAA, DMSO; then oxalic acid, Et2O, -78 °C; (b) MePPh3Br, KOC(CH3)3, THF; (c) BH3‚SMe2, THF, -78 to 0 °C, then
NaBO3‚H2O, 0 °C to room temperature; (d) NaH, MPMCl, Bu4NI, THF, ∆; (e) HCl, THF, H2O 0 °C to room temperature; (f) TPAP, NMO,
4 Å molecular sieves, CH2Cl2/CH3CN, 0 °C to room temperature; (g) ((CH3)2CH)2CHCH2CHO, LDA, THF, -78 °C; (h) MsCl, Et3N, 0 °C
to room temperature; (i) DBU, 0 °C to room temperature; (j) DDQ, H2O, CH2Cl2; (k) CH3(CH2)6C(O)Cl, pyridine, CH2Cl2, 0 °C; (l) BCl3,
CH2Cl2, -78 °C.
these compounds supports a common binding mode
where the only variable is the displacement of the sn-1
carbonyl.
of the conventional and novel PK-C isozymes bind
PDBU (phorbol 12,13-dibutyrate) with comparable af-
finities to those of the native isozymes.26
Although the modeling was performed with the
isolated C1b domain of the δ-isozyme, the measured Ki
values reflect the binding affinity of the compounds for
the complete R-isozyme. In principle, both the C1a and
C1b domains contain a binding pocket for phorbol/DAG.
However, studies with isolated domains have shown
that, with the exception of PK-Cγ, only the C1b domains
Con clu sion s
Complementing previous work from our laboratory
where we conclusively showed that each carbonyl on the
DAG-lactone is an indispensable pharmacophore,25 the
present findings confirm that the location of the side

648 J ournal of Medicinal Chemistry, 2004, Vol. 47, No. 3
Ta ble 1. Binding Affinities of Compounds 1-4
Tamamura et al.
room temperature overnight, then heated to reflux for 2 h,
cooled to room temperature, filtered through silica gel eluting
with hexane/EtOAc (3:2), and concentrated. Purification by
silica gel column chromatography gave 7 (46.9 g, 63% from
5). IR (neat) 2947 (CH), 2868 (CH), 1739 (CdO) cm-1; 1H NMR
(250 MHz, CDCl3) δ 1.17 (s, 9 H, (CH3)3CPh2SiOCH2C(O)CH2-
OCH2Ph), 4.43 (s, 4 H, (CH3)3CPh2SiOCH2C(O)CH2OCH2Ph),
4.64 (s, 2 H, (CH3)3CPh2SiOCH2C(O)CH2OCH2Ph), 7.41-7.72
(m, 15 H, (CH3)3CPh2SiOCH2C(O)CH2OCH2Ph); 13C NMR (62.9
MHz, CDCl3) δ 19.22, 26.76, 68.64, 73.18, 73.34, 127.82, 127.94,
128.40, 129.96, 132.33, 134.73, 135.40, 136.97, 206.51. Anal.
(C26H30O3Si‚0.9H2O) C, H.
Ki (nM)
Ki (nM)
compd
R
(Z-isomer)
(E isomer)
1
2
3
4
CH2OC(O)C9H19
CH2C(O)OC9H19
CH2CH2OC(O)C8H17
CH2CH2CH2OC(O)C7H15
2.3 ( 0.1a
55.7 ( 2.9
208 ( 27
736 ( 83
6.6 ( 1.0a
62.2 ( 7.2
459 ( 69
5-[(2,2-Dim eth yl-1,1-d ip h en yl-1-sila p r op oxy)m eth yl]-
5-[(ph en ylm eth oxy)m eth yl]-3,4,5-tr ih ydr ofu r an -2-on e (8).
Under argon, allylmagnesium bromide (205 mL, 1 M solution)
was added slowly to a solution of 7 (43 g, 107.7 mmol) in THF
(700 mL) at 0 °C, and the mixture was stirred overnight,
warming to room temperature. The resulting mixture was
quenched with 1 N aqueous HCl (100 mL), and the aqueous
layer was extracted with Et2O (3 × 200 mL). The combined
organic layers were dried over MgSO4, filtered, and concen-
trated. The crude product was then taken up in THF (105 mL)
under argon, cooled to -78 °C, and treated with BH3‚SMe2
(105 mL, 2 M solution). After 1 h, the reaction mixture was
warmed to room temperature and stirred overnight. The
reaction mixture was concentrated and then sequentially
treated with CH2Cl2 (1.1 L), 4 Å molecular sieves (53 g), and
PCC (226 g, 1050 mmol), which was added in portions. The
resulting mixture was then stirred for 2 days, after which it
was filtered through silica gel eluting with hexane/EtOAc (3:
2) and concentrated. Purification by silica gel column chro-
matography gave 8 (31.7 g, 62% for three steps). IR (neat) 3058
(CH), 2936 (CH), 2861 (CH), 1777 (CdO) cm-1; 1H NMR (250
MHz, CDCl3) δ 1.16 (s, 9 H, (CH3)3CPh2SiOCH2C), 2.27 (app
t, J ) 8.7 Hz, 2 H, lactone â-CH2), 2.65-2.80 (m, 2 H, lactone
R-CH2), 3.66 (AB quartet, J ) 10.4 Hz, 2 H, PhCH2OCH2C),
3.81 (AB quartet, J ) 10.9 Hz, 2 H, (CH3)3CPh2SiOCH2C), 4.63
(s, 2 H, PhCH2OCH2C), 7.34-7.75 (m, 15 H, PhCH2OCH2C
and (CH3)3CPh2SiOCH2C); 13C NMR (62.9 MHz, CDCl3) δ
19.20, 26.01, 26.72, 29.20, 66.55, 72.34, 73.61, 87.36, 127.47,
127.65, 127.72, 128.31, 129.79, 132.41, 132.64, 135.44, 135.50,
137.53, 176.93; FAB-MS (m/z, relative intensity) 475 (MH+,
5), 91 (100). Anal. (C29H34O4Si‚0.25H2O) C, H.
5-[(2,2-Dim eth yl-1,1-d ip h en yl-1-sila p r op oxy)m eth yl]-
5-[(p h en ylm eth oxy)m eth yl]oxola n -2-ol (9). Under argon,
DIBAL (4.6 mL, 4.59 mmol, 1 M in THF) was added to a
mixture of 8 (1.81 g, 3.82 mmol) in toluene (25 mL) at -78 °C,
and the mixture was stirred for 2 h. An additional equivalent
of DIBAL was added to push the reaction to completion, after
which it was quenched with methanol (493 µL). The reaction
mixture was diluted with CH2Cl2 (50 mL), and then Celite was
added followed by a saturated solution of aqueous ammonium
chloride (3 mL). The mixture was dried over MgSO4, filtered,
and concentrated to give 9 as an oil (1.6 g, 88%), which was
used in the next step without further purification.
(2,2-Dim eth yl-1,1-d ip h en yl-1-sila p r op oxy){5-m eth oxy-
2-[(p h en ylm et h oxy)m et h yl]oxola n -2-yl}m et h a n e (10).
Dowex 50WX8-200 (13.4 g), methanol (34 mL), and AcOH (63
mL, 80%) were added in portions to a stirring solution of 9
(crude, 1.6 g, 3.27 mmol) in THF (15 mL), and the reaction
was monitored by TLC. Upon completion, the mixture was
filtered, diluted with toluene, and concentrated. The resulting
mixture was neutralized with NaHCO3, dried over MgSO4,
filtered, and concentrated. Purification by silica gel column
chromatography (5% EtOAc in hexane) gave an anomeric
mixture of 10A (376 mg, 23%) and 10B (406 mg, 25%) as oils.
When feasible, the anomers were separated for characteriza-
tion.
1430 ( 140
a
From ref 20.
chain carbonyl in a DAG-lactone has to have perfect
mimicry to the sn-1 carbonyl of the parent DAG for it
to display high binding affinity. A proposed model from
this work is that the missing pharmacophore in the
ternary complex, which includes the membrane, is close
to the membrane-protein interface. This is in agree-
ment with the recent proposal of Newton et al. suggest-
ing that the two C1 domains are likely to be oriented
with their ligand-binding pockets facing the mem-
brane.27
Exp er im en ta l Section
Biologica l Activity. Enzyme-ligand interactions were as-
sessed in terms of the ability of the ligand to displace bound
[20-3H]phorbol 12,13-dibutyrate (PDBU) from a recombinant
single isozyme (PKC-R) in the presence of phosphatidyl-
serine.28-31 The octanol/water partition coefficients (log P) were
calculated according to the fragment-based program KOWWIN
1.67 (http://www.epa.gov/oppt/exposure/docs/episuitedl.htm).
Gen er a l P r oced u r es. All chemical reagents were com-
mercially available. DCAA ) dichloroacetic acid; DCC ) 1,3-
dicyclohexylcarbodiimide; DDQ ) 2,3-dichloro-5,6-dicyano-1,4-
benzoquinone; DIBAL ) diisobutylaluminum hydride; MPM
) methoxyphenylmethyl; MsCl ) methanesulfonyl chloride;
PCC ) pyridinium chlorochromate; NMO ) 4-methylmorpho-
line N-oxide; TBDPSCl ) tert-butyldiphenylsilyl chloride;
TPAP ) tetrapropylammonium perruthenate. Column chro-
matography was performed on silica gel 60, 230-400 mesh
(E. Merck). 1H and 13C NMR spectra were recorded on a Bruker
AC-250 instrument at 250 and 62.9 MHz, respectively. Spectra
are referenced to the solvent in which they were run (7.24 ppm
for CDCl3). Infrared spectra were recorded on a Perkin-Elmer
1600 series FT-IR. Positive ion fast-atom bombardment mass
spectra (FAB-MS) were obtained on a VG 7070E mass spec-
trometer at an accelerating voltage of 6 kV and a resolution
of 2000. Glycerol was used as the sample matrix, and ioniza-
tion was effected by a beam of xenon atoms. Elemental
analyses were performed by Atlantic Microlab, Inc., Atlanta,
GA.
3-(2,2-Dim eth yl-1,1-d ip h en yl-1-sila p r op oxy)-1-(p h en yl-
m eth oxy)p r op a n -2-ol (6). Under argon, a solution of com-
mercially available 3-benzyloxy-1,2-propanediol (5, 32.5 g, 178
mmol) in CH2Cl2 (557 mL) was treated with imidazole (27.9
g, 410 mmol) and stirred for 85 min at room temperature. The
mixture was then diluted sequentially with CH2Cl2 (279 mL)
and DMF (279 mL), treated with tert-butyldiphenylsilyl chlo-
ride (53.7 mL, 207 mmol) and stirred at room temperature
overnight. The reaction mixture was then diluted with Et2O
(1 L) and quenched with saturated aqueous NH4Cl (200 mL).
The organic layer was washed sequentially with H2O and
brine, dried over MgSO4, and concentrated to give 6, which
was used without further purification.
10A: IR (neat) 3018 (CH), 2932 (CH), 2859 (CH) 1104 (CO)
cm-1; H NMR (250 MHz, CDCl3) δ 1.15 (s, 9 H, (CH3)3CPh2-
1
3-(2,2-Dim eth yl-1,1-d ip h en yl-1-sila p r op oxy)-1-(p h en yl-
m eth oxy)a ceton e (7). Under argon, a solution of 6 (178
mmol) in CH2Cl2 (1 L) was successively treated with 4 Å
molecular sieves (89 g) and PCC (154 g, 714 mmol), stirred at
SiOCH2C), 1.86-2.15 (m, 4 H, ring-CH2CH2), 3.41 (s, 3 H,
>CHOCH3), 3.57-3.92 (m, 4 H, PhCH2OCH2C and (CH3)3-
CPh2SiOCH2C), 4.68 (s, 2 H, PhCH2OCH2C), 5.07 (d, J ) 2.9
Hz, 1 H, >CHOCH3), 7.34-7.51 (m, 10 H, (CH3)3CPh2-

Analogues of Diacylglycerol
J ournal of Medicinal Chemistry, 2004, Vol. 47, No. 3 649
SiOCH2C), 7.75-7.80 (m, 5 H, PhCH2OCH2C); 13C NMR (62.9
MHz, CDCl3) δ 19.27, 26.79, 28.15, 32.11, 54.27, 65.42, 73.41,
74.24, 86.46, 105.66, 127.26, 127.46, 127.49, 128.11, 129.42,
133.35, 133.45, 135.50, 135.53, 138.36; FAB-MS (m/z, relative
intensity) 459 (MH+ - CH3OH, 4), 91 (100). Anal. (C30H38O4-
Si) C, H.
>CHOCH3), 3.56-3.64 (m, 2 H, PhCH2OCH2C), 4.63-4.74 (m,
2 H, PhCH2OCH2C), 5.16-5.41 (m, 3 H, CCHdCH2 and
>CHOCH3), 6.03 (dd, J ) 17.1, 10.7 Hz, 1 H, CCHdCH2),
7.35-7.43 (m, 5 H, PhCH2OCH2C); 13C NMR (62.9 MHz,
CDCl3) δ 31.17, 31.37, 54.37, 73.37, 76.63, 86.14, 105.48,
113.00, 127.32, 127.45, 128.13, 138.31, 140.17; FAB-MS (m/z,
relative intensity) 217 (MH+ - CH3OH, 19), 91 (100). Anal.
(C15H20O3) C, H.
10B: IR (neat) 3017 (CH), 2932 (CH), 2860 (CH), 1102 (CO)
1
cm-1; H NMR (250 MHz, CDCl3) δ 1.16 (s, 9 H, (CH3)3CPh2-
SiOCH2C), 1.95-2.10 (m, 4 H, ring-CH2CH2), 3.33 (s, 3 H,
>CHOCH3), 3.61-3.91 (m, 4 H, PhCH2OCH2C and (CH3)3-
CPh2SiOCH2C), 4.68 (s, 2 H, PhCH2OCH2C), 5.13 (d, J ) 4.2
Hz, 1 H, >CHOCH3), 7.37-7.51 (m, 10 H, (CH3)3CPh2-
SiOCH2C), 7.74-7.81 (m, 5 H, PhCH2OCH2C); 13C NMR (62.9
MHz, CDCl3) δ 19.31, 26.81, 28.55, 32.35, 54.32, 67.59, 72.38,
73.52, 86.44, 105.68, 127.29, 127.40, 127.47, 128.15, 129.44,
133.43, 133.45, 135.49, 135.54, 138.37; FAB-MS (m/z, relative
intensity): 459 (MH+ - CH3OH, 7), 91 (100). Anal. (C30H38O4-
Si) C, H.
{5-Met h oxy-2-[(p h en ylm et h oxy)m et h yl]oxola n -2-yl}-
m eth a n -1-ol (11). A solution of 10 (742 mg, 1.5 mmol, mixture
of anomers) in THF (15 mL) was treated with TBAF (2.3 mmol,
1.5 equiv, 1 M in THF) at 0 °C for 1 h, after which additional
TBAF (2.3 mmol, 1.5 equiv, 1 M in THF) was added. After a
total time of 2 h, the reaction mixture was warmed to room
temperature and stirred for 1 h more. The crude reaction
mixture was then concentrated in vacuo and purified by silica
gel column chromatography to give an anomeric mixture of
11A (46%) and 11B (53%) as colorless oils. When feasible, the
anomers were separated for characterization.
11A: IR (neat) 3467 (OH), 3009 (CH), 2930 (CH) 2865 (CH),
1095 (CO), 1034 (CO) cm-1; 1H NMR (250 MHz, CDCl3) δ 1.89-
2.17 (m, 4 H, ring-CH2CH2), 3.45-3.80 (m, 4 H, PhCH2OCH2C
and HOCH2C), 3.46 (s, 3 H, >CHOCH3), 4.63 (AB quartet, J
) 12.3 Hz, 2 H, PhCH2OCH2C), 5.10 (d, J ) 3.7 Hz, 1 H,
>CHOCH3), 7.34-7.43 (m, 5 H, PhCH2OCH2C); 13C NMR (62.9
MHz, CDCl3) δ 26.92, 32.61, 54.97, 66.53, 73.10, 73.49, 87.12,
105.90, 127.41, 127.53, 128.25, 137.91; FAB-MS (m/z, relative
intensity): 221 (MH+ - CH3OH, 56), 91 (100). Anal. (C14H20O4)
C, H.
2-{5-Meth oxy-2-[(p h en ylm eth oxy)m eth yl]oxola n -2-yl}-
eth a n -1-ol (14). A solution of BH3‚SMe2 in THF (812 µL, 1.63
mmol, 2 M solution) was added dropwise to a -78 °C solution
of 13 (202 mg, 0.81 mmol) in THF (3 mL). The reaction mixture
was then slowly allowed to reach 0 °C and stirred. After 17 h
the reaction mixture was treated with NaBO3‚H2O (250 mg,
1.63 mmol) at 0 °C and then stirred for 2 h, warming to room
temperature. The reaction mixture was diluted with EtOAc,
the organic layer was washed with water and brine, dried over
MgSO4, and concentrated. Purification by silica gel column
chromatography (EtOAc/hexanes) gave an anomeric mixture
of 14 (121 mg, 56%) as a colorless oil. IR (neat) 3504 (OH),
3017 (CH), 2931 (CH), 1097 (CO) 1034 (CO) cm-1; FAB-MS
(m/z, relative intensity) 235 (MH+ - CH3OH, 20), 91 (100).
Anal. (C15H22O4) C, H.
1-[(4-Meth oxyp h en yl)m eth oxy]-2-{5-m eth oxy-2-[(p h e-
n ylm eth oxy)m eth yl]oxola n -2-yl}eth a n e (15). According to
the literature,33 a solution of 14 (110 mg, 0.41 mmol) in THF
(2 mL) was added dropwise to a 0 °C slurry of NaH (33 mg,
0.82 mmol, 60% in oil) in THF (2 mL). The reaction mixture
was then heated to reflux, and p-methoxybenzyl chloride (141
µL) and Bu4NI (30 mg) were added in sequential portions over
the course of 3 days as the reaction was monitored by TLC.
The reaction mixture was then cooled to 0 °C, and quenched
with methanol (500 µL) followed by saturated aqueous NH4-
Cl (10 mL). The aqueous solution was then extracted with CH2-
Cl2, dried over MgSO4, and concentrated. Purification by silica
gel column chromatography (hexanes/EtOAc) gave an ano-
meric mixture of 15 (122 mg, 77%) as an oil. IR (neat) 3017
(CH), 2931 (CH), 2862 (CH), 1097 (CO) 1036 (CO) cm-1; FAB-
MS (m/z, relative intensity) 387 (MH+, 2), 121 (100). Anal.
(C23H30O5) C, H.
5-{2-[(4-Met h oxyp h en yl)m et h oxy]et h yl}-5-[(p h en yl-
m eth oxy)m eth yl]oxola n -2-ol (16). According to the litera-
ture,33 HCl (350 µL, 2 N) was added to a 0 °C solution of 15 in
THF (4.7 mL) and H2O (1.8 mL). After the mixture was allowed
to warm to room temperature over a 5 h period, additional
HCl (400 µL, 2 N) was added and the reaction mixture was
stirred overnight. The reaction was then quenched with solid
NaHCO3 followed by H2O (10 mL). The aqueous solution was
extracted with CH2Cl2, dried over MgSO4, and concentrated.
Purification by silica gel column chromatography (hexane/
EtOAc) gave an anomeric mixture of 16 (69 mg, 74%) as a
colorless oil. IR (neat) 3419 (OH), 3010 (CH), 2956 (CH), 2865
(CH), 1093 (CO) 1035 (CO) cm-1; FAB-MS (m/z, relative
intensity) 371 (MH+ - H2, 3), 355 (MH+ - H2O, 6), 121 (100).
Anal. (C22H28O5) C, H.
11B: IR (neat) 3587 (OH), 3016 (CH), 2925 (CH) 2867 (CH),
1098 (CO), 1035 (CO) cm-1; 1H NMR (250 MHz, CDCl3) δ 1.92-
2.10 (m, 4 H, ring-CH2CH2), 3.38 (s, 3 H, >CHOCH3), 3.56-
3.79 (2 overlapping AB quartets, 4 H, PhCH2OCH2C and
HOCH2C), 4.65 (AB quartet, J ) 12.2 Hz, 2 H, PhCH2OCH2C),
5.11 (t, J ) 1.7 Hz, 1 H, >CHOCH3), 7.34-7.43 (m, 5 H,
PhCH2OCH2C); 13C NMR (62.9 MHz, CDCl3) δ 28.70, 32.46,
54.43, 66.27, 73.51, 75.09, 85.46, 105.80, 127.48, 127.54,
128.25, 137.93; FAB-MS (m/z, relative intensity) 221 (MH+
CH3OH, 54), 91 (100). Anal. (C14H20O4) C, H.
-
{5-Met h oxy-2-[(p h en ylm et h oxy)m et h yl]oxola n -2-yl}-
for m a ld eh yd e (12). DCC (1.10 g, 5.33 mmol) and DCAA (54.9
µL, 0.67 mmol) were added to a solution of 11 (336 mg, 1.33
mmol) in DMSO (2.7 mL) and stirred overnight. After 16 h
the reaction was quenched with portions of oxalic acid (480
mg, 5.3 mmol), diluted with Et2O (20 mL), and cooled to -78
°C. The resulting mixture was filtered and washed with cold
Et2O to remove excess DCU. The organic solution was then
washed with water and brine, dried over MgSO4, and concen-
trated to give 12 as a mixture of anomers that was used
directly without further purification.
5-{2-[(4-Met h oxyp h en yl)m et h oxy]et h yl}-5-[(p h en yl-
m eth oxy)m eth yl]-3,4,5-tr ih yd r ofu r a n -2-on e (17). Accord-
ing to the literature,34 solid TPAP (2.2 mg, 0.006 mmol, 5 mol
%) was added in one portion to a stirring mixture of 16 (46.5
mg, 0.125 mmol), NMO (21.0 mg, 0.19 mmol), and powdered
4 Å molecular sieves (55 mg heat-dried under vacuum) in CH2-
Cl2/CH3CN (0.23 mL, 10% CH3CN in CH2Cl2) at 0 °C under
argon. The reaction mixture was stirred overnight and allowed
to reach room temperature. The crude reaction mixture was
then concentrated in vacuo, filtered through Celite (eluting
with CH2Cl2) followed by filtration through silica gel (eluting
with EtOAc), and concentrated to give 17 (38.9 mg, 84%). IR
(5-Me t h o x y -2-v in y lo x o la n -2-y l)(p h e n y lm e t h o x y )-
m eth a n e (13). According to the literature,29,32 potassium tert-
butoxide (2.66 mL, 2.66 mmol, 1.0 M in THF) was combined
with MePPh3Br (952 mg, 2.66 mmol) and stirred. After 30 min
the mixture was added slowly to a solution of 12 (1.33 mmol)
in THF (3.3 mL), and the resulting mixture was stirred
overnight. The reaction was quenched with AcOH (0.2 mL),
and the mixture was filtered and concentrated. Purification
by silica gel column chromatography (EtOAc/hexane) gave an
anomeric mixture of 13 (240 mg, 73%) as a colorless oil. When
feasible, the anomers were separated for characterization.
On e An om er : IR (neat) 3017 (CH), 2919 (CH), 2859 (CH),
(neat) 3020 (CH), 2928 (CH), 2862 (CH), 1766 (CdO) cm-1
;
1H NMR (250 MHz, CDCl3) δ 2.05-2.10 (m, 2 H, OCH2CH2C),
2.23-2.78 (m, 4 H, lactone-CH2CH2), 3.59-3.68 (m, 4 H, OCH2-
CH2C and PhCH2OCH2C), 3.89 (s, 3 H, CH3OC6H4CH2O), 4.47
(s, 2 H, CH3OC6H4CH2O), 4.55-4.67 (m, 2 H, PhCH2OCH2C),
6.92-6.97 (m, 2 H, CH3OC6H4CH2O), 7.26-7.43 (m, 7 H, CH3-
OC6H4CH2O and PhCH2OCH2C); 13C NMR (62.9 MHz, CDCl3)
1
1645 (CdC), 1097 (CO), 1033 (CO) cm-1; H NMR (250 MHz,
CDCl3) δ 1.81-2.16 (m, 4 H, ring-CH2CH2), 3.43 (s, 3 H,

650 J ournal of Medicinal Chemistry, 2004, Vol. 47, No. 3
Tamamura et al.
δ 29.26, 29.41, 36.86, 55.19, 65.05, 72.77, 73.50, 74.98, 86.60,
113.70, 127.40, 127.61, 128.30, 129.16, 129.91, 137.63, 159.08,
177.11; FAB-MS (m/z, relative intensity) 371 (MH+, 4), 121
(100). Anal. (C22H26O5) C, H.
(CH(CH3)2)2), 1.80-1.95 (m, 2 H, CdCHCH2CH(CH(CH3)2)2),
1.95 (br s, 1 H, HOCH2CH2C), 2.07-2.20 (m, 4 H, HOCH2CH2C
and CdCHCH2CH(CH(CH3)2)2), 2.71-3.02 (m, 2 H, lactone-
CH2), 3.56 (AB quartet, J ) 10.2 Hz, 2 H, PhCH2OCH2C), 3.87
(t, J ) 6.0 Hz, 2 H, HOCH2CH2C), 4.64 (s, 2 H, PhCH2OCH2C),
6.80-6.90 (m, 1 H, CdCHCH2CH(CHMe2)2), 7.34-7.44 (m, 5
H, PhCH2OCH2C); 13C NMR (62.9 MHz, CDCl3) δ 19.25, 19.31
21.51, 21.57, 28.58, 29.12, 29.15, 33.87 39.94, 50.20, 57.91,
73.62, 74.10, 83.49, 125.40, 127.52, 127.73, 128.32, 137.24,
142.87, 170.10; FAB-MS (m/z, relative intensity) 375 (MH+,
44), 91 (100). Anal. (C23H34O4) C, H.
5-{2-[(4-Meth oxyp h en yl)m eth oxy]eth yl}-3-[4-m eth yl-3-
(m eth yleth yl)p en tylid en e]-5-[(p h en ylm eth oxy)m eth yl]-
4,5-d ih yd r ofu r a n -2-on e (18E/Z). Under argon, a -78 °C
solution of 17 (1.0 g, 2.81 mmol) in THF (10 mL) was treated
dropwise with LDA (2.0 mL, 3.93 mmol, 2 M solution in
heptane/THF/ethylbenzene) and stirred for 2 h. A solution of
4-methyl-3-(methylethyl)pentanal (719 mg, 5.1 mmol) in THF
(10 mL) was added dropwise maintaining the same temper-
ature, and the reaction was monitored by TLC. The reaction
was quenched with saturated aqueous NH4Cl and then
warmed to room temperature. The aqueous layer was ex-
tracted with Et2O, washed with water, dried, and concentrated
to give an oil that was then taken up in CH2Cl2 (20 mL) under
argon and treated with MsCl (0.44 mL, 5.62 mmol) and Et3N
(1.57 mL, 11.23 mmol) at 0 °C. The reaction mixture was
stirred at 0 °C for 30 min and then at room temperature for 2
h. The reaction mixture was then cooled to 0 °C, DBU (2.1
mL, 14.0 mmol) was added, and the resultant solution was
stirred at room temperature for 3 h. The crude reaction
mixture was filtered through a pad of silica gel and concen-
trated. Purification by silica gel column chromatography gave
18E (600 mg, 43%) and 18Z (407 mg, 29%).
(Z)-5-(2-H yd r oxyet h yl)-3-[4-m et h yl-3-(m et h ylet h yl)-
p en tylid en e]-5-[(p h en ylm eth oxy)m eth yl]-4,5-d ih yd r ofu -
r a n -2-on e (19Z). According to the literature,32 DDQ (187 mg,
0.83 mmol) was added to a stirring solution of 18Z (271 mg,
0.55 mmol) in CH2Cl2 (3.8 mL) and H2O (0.77 mL). After 2 h,
the mixture was filtered through Celite and washed with CH2-
Cl2. The organic phase was washed sequentially with saturated
aqueous NaHCO3 and brine, dried, and concentrated. Purifica-
tion by chromatography gave 19Z (1.88 mg, 92%) as an oil. IR
(neat) 3608 (OH), 3022 (CH), 2960 (CH), 2872 (CH), 1749
1
(CdO), 1666 (CdC) cm-1; H NMR (250 MHz, CDCl3) δ 0.92,
0.93, 0.97, and 0.98 (d, J ) 6.8 Hz, 3 H, CdCHCH2CH(CH-
(CH3)2)2), 1.16 (pentet, J ) 5.4 Hz, 1 H, CdCHCH2CH(CH-
(CH3)2)2), 1.75-1.93 (m, 2 H, CdCHCH2CH(CH(CH3)2)2), 2.05
(t, J ) 6.1 Hz, 2 H, HOCH2CH2C), 2.43 (s, 1 H, HOCH2CH2C),
2.75-3.05 (m, 4 H, lactone-CH2 and CdCHCH2CH(CH-
(CH3)2)2), 3.58 (s, 2 H, PhCH2OCH2C), 3.83 (t, J ) 6.1 Hz, 2
H, HOCH2CH2C), 4.63 (s, 2 H, PhCH2OCH2C), 6.15-6.30 (m,
1 H, CdCHCH2CH(CH(CH3)2)2), 7.35-7.41 (m, 5 H, PhCH2-
OCH2C); 13C NMR (62.9 MHz, CDCl3) δ 19.36, 19.39, 21.59,
21.62, 26.18, 29.29, 37.23, 39.73, 51.10, 57.78, 73.55, 73.95,
82.92, 123.49, 127.51, 127.70, 128.30, 137.34, 146.43, 169.05;
FAB-MS (m/z, relative intensity) 375 (MH+, 22), 91 (100). Anal.
(C23H34O4) C, H.
18E: IR (neat) 3022 (CH), 2961 (CH), 2873 (CH), 1746
1
(CdO), 1676 (CdC) cm-1; H NMR (250 MHz, CDCl3) δ 0.90,
(d, J ) 6.8 Hz, 6 H, CdCHCH2CH(CH(CH3)2)2), 0.95 and 0.96
(d, J ) 6.8 Hz, 3 H, CdCHCH2CH(CH(CH3)2)2), 1.29-1.32 (m,
1 H, CdCHCH2CH(CH(CH3)2)2), 1.75-1.93 (m, 2 H, CdCHCH2-
CH(CH(CH3)2)2), 2.02-2.20 (m, 2 H, OCH2CH2C), 2.75-3.02
(m, 2 H, CdCHCH2CH(CH(CH3)2)2), 2.94 (s, 2 H, lactone-CH2),
3.57 (s, 2 H, PhCH2OCH2C), 3.62-3.67 (m, 2 H, OCH2CH2C),
3.87 (s, 3 H, CH3OC6H4CH2O), 4.44 (s, 2 H, CH3OC6H4CH2O),
4.60 (app d, J ) 1.7 Hz, 2 H, PhCH2OCH2C), 6.76-6.85 (m, 1
H, CdCHCH2CH(CH(CH3)2)2), 6.91-6.95 (m, 2 H, PhCH2-
OCH2C), 7.23-7.45 (m, 7 H, PhCH2OCH2C and CH3OC6H4-
CH2O); 13C NMR (62.9 MHz, CDCl3) δ 19.22, 19.32, 21.47,
21.57, 28.48, 29.10, 29.15, 33.39, 36.91, 50.17, 55.16, 65.00,
72.74, 73.43, 74.39, 83.48, 113.64, 126.13, 127.37, 127.51,
128.20, 129.11, 129.98, 137.65, 141.95, 159.02, 170.42; FAB-
MS (m/z, relative intensity) 495 (MH+, 4), 121 (100). Anal.
(C31H45O5‚H2O) C, H.
(E)-3-[4-Meth yl-3-(m eth yleth yl)p en tylid en e]-5-(2-oxo-
et h yl)-5-[(p h en ylm et h oxy)m et h yl]-4,5-d ih yd r ofu r a n -2-
on e (20E). Under argon, a solution of DMSO (107 µL) in
CH2Cl2 (0.20 mL) was added dropwise to a -78 °C solution of
oxalyl chloride (60 µL) in CH2Cl2 (0.70 mL). After 20 min, a
solution of 19E (172 mg, 0.46 mmol) in CH2Cl2 (0.20 mL) was
added and the mixture was stirred at -78 °C for 1 h.
Triethylamine (0.38 mL) was added dropwise, and the result-
ing mixture was stirred for 2 h warming to 0 °C. The reaction
mixture was then again cooled to -78 °C, quenched with
saturated aqueous NH4Cl, warmed to room temperature, and
extracted with Et2O. The organic layer was washed with H2O,
dried over MgSO4, and concentrated to give 20E, which was
used directly in the next step without further purification.
(Z)-3-[4-Meth yl-3-(m eth yleth yl)p en tylid en e]-5-(2-oxo-
et h yl)-5-[(p h en ylm et h oxy)m et h yl]-4,5-d ih yd r ofu r a n -2-
on e (20Z). Under argon, a solution of DMSO (64 µL) in CH2Cl2
(0.12 mL) was added dropwise to a -78 °C solution of oxalyl
chloride (36 µL) in CH2Cl2 (0.42 mL). After 20 min, a solution
of 19Z (103 mg, 0.27 mmol) in CH2Cl2 (0.12 mL) was added
and the mixture was stirred at -78 °C for 1 h. Triethylamine
(0.23 mL) was added dropwise, and the resulting mixture was
stirred for 2 h warming to 0 °C. The reaction mixture was then
again cooled to -78 °C, quenched with saturated aqueous NH4-
Cl, warmed to room temperature, and extracted with Et2O.
The organic layer was washed with H2O, dried over MgSO4,
and concentrated to give 20Z, which was used directly in the
next step without further purification.
18Z: IR (neat) 3019 (CH), 2960 (CH), 2870 (CH), 1746
1
(CdO), 1666 (CdC) cm-1; H NMR (250 MHz, CDCl3) δ 0.94,
0.95, 0.98, and 0.99 (d, J ) 6.8 Hz, 3 H, CdCHCH2CH(CH-
(CH3)2)2), 1.17 (pentet, J ) 5.4 Hz, 1 H, CdCHCH2CH(CH-
(CH3)2)2), 1.75-1.95 (m, 2 H, CdCHCH2CH(CH(CH3)2)2), 2.08-
2.13 (m, 2 H, OCH2CH2C), 2.78-2.86 (m, 2 H, lactone-CH2),
2.90-2.96 (m, 2 H, CdCHCH2CH(CH(CH3)2)2), 3.56 (s, 2 H,
PhCH2OCH2C), 3.65 (t, J ) 6.3 Hz, 2 H, OCH2CH2C), 3.88 (s,
3 H, CH3OC6H4CH2O), 4.45 (s, 2 H, CH3OC6H4CH2O), 4.61 (s,
2 H, CCH2OCH2Ph), 6.22 (tt, J ) 7.3, 2.0 Hz, 1 H, CdCHCH2-
CH(CH(CH3)2)2), 6.88-7.00 (m, 2 H, CH3OC6H4CH2O), 7.25-
7.48 (m, 7 H, CH3OC6H4CH2O and PhCH2OCH2C); 13C NMR
(62.9 MHz, CDCl3) δ 19.38, 19.41, 21.60, 21.63, 26.13, 29.30,
36.72, 36.81, 51.13, 55.17, 65.06, 72.71, 73.42, 74.21, 82.80,
113.66, 124.09, 127.41, 127.52, 128.22, 129.09, 130.05, 137.74,
145.66, 159.03, 169.24; FAB-MS (m/z, relative intensity) 493
(MH+ - H2, 4), 121 (100). Anal. (C31H45O5) C, H.
(E)-5-(2-H yd r oxyet h yl)-3-[4-m et h yl-3-(m et h ylet h yl)-
p en tylid en e]-5-[(p h en ylm eth oxy)m eth yl]-4,5-d ih yd r ofu -
r a n -2-on e (19E). According to the literature,29,32 DDQ (122
mg, 0.54 mmol) was added to a stirring solution of 18E (177
mg, 0.36 mmol) in CH2Cl2 (2.5 mL) and H2O (0.50 mL). After
2.5 h, the mixture was filtered through Celite and washed with
CH2Cl2. The organic phase was washed sequentially with
saturated aqueous NaHCO3 and brine, dried, and concen-
trated. Purification by chromatography gave 19E (101 mg,
76%) as an oil. IR (neat) 3466 (OH), 3017 (CH), 2961 (CH),
2873 (CH), 1748 (CdO), 1675 (CdC) cm-1; 1H NMR (250 MHz,
CDCl3) δ 0.91, 0.92, 0.97, and 0.98 (d, J ) 6.8 Hz, 3 H,
CdCHCH2CH(CH(CH3)2)2), 1.25-1.35 (m, 1 H, CdCHCH2CH-
(E)-2-{4-[4-Meth yl-3-(m eth yleth yl)p en tylid en e]-5-oxo-
2-[(ph en ylm eth oxy)m eth yl]-2-2,3-dih ydr ofu r yl}acetic Acid
(21E). Under argon, 2-methyl-2-butene (320 µL), NaClO2 (67
mg, 80%), and a solution of NaH2PO4 (71 mg) in H2O (460 µL)
were added to a solution of 20E (0.46 mmol) in tert-butyl
alcohol (580 µL), and the mixture was stirred overnight at
room temperature. The mixture was then concentrated, diluted
with H2O, acidified to pH 2, and extracted three times with
Et2O. The combined ethereal layers were dried over MgSO4
and concentrated to give 21E, which was used directly in the
next step without further purification.

Analogues of Diacylglycerol
J ournal of Medicinal Chemistry, 2004, Vol. 47, No. 3 651
(CH), 1751 (CdO), 1674 (CdC) cm-1 1H NMR (250 MHz,
(Z)-2-{4-[4-Meth yl-3-(m eth yleth yl)p en tylid en e]-5-oxo-
2-[(ph en ylm eth oxy)m eth yl]-2-2,3-dih ydr ofu r yl}acetic Acid
(21Z). Under argon, 2-methyl-2-butene (192 µL), NaClO2 (40
mg, 80%), and a solution of NaH2PO4 (43 mg) in H2O (367 µL)
were added to a solution of 20Z (0.27 mmol) in tert-butyl
alcohol (347 µL), and the mixture was stirred overnight at
room temperature. The mixture was then concentrated, diluted
with H2O, acidified to pH 2, and extracted three times with
Et2O. The combined ethereal layers were dried over MgSO4
and concentrated to give 21Z, which was used directly in the
next step without further purification.
;
CDCl3) δ 0.91-1.00 (m, 15 H, CdCHCH2CH(CH(CH3)2)2 and
CH3(CH2)6CH2CH2OC(O)CH2C), 1.24-1.45 (m, 13 H, CdCH-
CH2CH(CH(CH3)2)2 and CH3(CH2)6CH2CH2OC(O)CH2C), 1.61-
1.76 (m, 2 H, CH3(CH2)6CH2CH2OC(O)CH2C), 1.77-1.96 (m,
2 H, CdCHCH2CH(CH(CH3)2)2), 2.14-2.26 (m, 2 H, lactone-
CH2), 2.40 (br s, 1 H, HOCH2C), 2.84 (d, J ) 15.9 Hz, 2 H,
CdCHCH2CH(CH(CH3)2)2), 2.90-3.00 (m, 2 H, CH3(CH2)6CH2-
CH2OC(O)CH2C), 3.79 (AB quartet, J ) 12.2 Hz, 2 H,
HOCH2C), 4.15 (t, J ) 6.8 Hz, 2 H, CH3(CH2)6CH2CH2OC(O)-
CH2C), 6.83-6.92 (m, 1 H, CdCHCH2CH(CH(CH3)2)2); 13C
NMR (62.9 MHz, CDCl3) δ 14.05, 19.26 19.30, 21.55, 22.59,
25.80, 28.39, 28.62, 29.15, 29.38, 31.77, 32.16, 40.47, 50.22,
65.18, 67.23, 82.52, 125.14, 143.56, 169.32, 169.82; FAB-MS
(m/z, relative intensity) 425 (MH+, 100). Anal. (C25H44O5) C,
H.
(E)-Non yl 2-{4-[4-Meth yl-3-(m eth yleth yl)p en tylid en e]-
5-oxo-2-[(p h en ylm et h oxy)m et h yl]-2-2,3-d ih yd r ofu r yl}-
a ceta te (22E). Under argon, 1-nonanol (400 µL), DMAP (5.60
mg), and DCC (472 mg) were added to a 0 °C solution of 21E
(0.458 mmol) in pyridine (1 mL), and the mixture was stirred
overnight warming to room temperature. The crude reaction
mixture was filtered through Celite and concentrated. Puri-
fication by silica gel column chromatography gave 22E (110
mg, 47% for three steps from 19E) as a colorless oil. IR (neat)
3020 (CH), 2959 (CH), 2930 (CH), 2858 (CH), 1751 (CdO),
1676 (CdC) cm-1; 1H NMR (250 MHz, CDCl3) δ 0.90-0.99 (m,
15 H, CdCHCH2CH(CH(CH3)2)2 and CH3(CH2)6CH2CH2OC-
(O)CH2C), 1.23-1.45 (m, 13 H, CdCHCH2CH(CH(CH3)2)2 and
CH3(CH2)6CH2CH2OC(O)CH2C), 1.58-1.74 (m, 2 H, CH3-
(CH2)6CH2CH2OC(O)CH2C), 1.75-1.94 (m, 2 H, CdCHCH2CH-
(CH(CH3)2)2), 2.15-2.20 (m, 2 H, CdCHCH2CH(CH(CH3)2)2),
2.88 (AB quartet, J ) 15.9 Hz, 2 H, CH3(CH2)6CH2CH2OC-
(O)CH2C, 2.98 (br s, 2 H, lactone-CH2), 3.65 (s, 2 H, PhCH2-
OCH2C), 4.12 (t, J ) 6.8 Hz, 2 H, CH3(CH2)6CH2CH2OC(O)-
CH2C), 4.63 (s, 2 H, PhCH2OCH2C), 6.80-6.89 (m, 1 H,
CdCHCH2CH(CH(CH3)2)2), 7.35-7.41 (m, 5 H, PhCH2OCH2C);
13C NMR (62.9 MHz, CDCl3) δ 14.06, 19.23 19.33, 21.49, 21.58,
22.60, 25.81, 28.40, 28.55, 29.10, 29.17, 29.39, 31.77, 32.89,
41.03, 50.19, 65.00, 73.52, 74.21, 81.49, 125.46, 127.41, 127.61,
128.24, 137.43, 142.56, 169.13, 169.78; FAB-MS (m/z, relative
intensity) 515 (MH+, 18), 91 (100). Anal. (C32H50O5) C, H.
(Z)-Non yl 2-{2-(Hyd r oxym eth yl)-4-[4-m eth yl-3-(m eth -
ylet h yl)p en t ylid en e]-5-oxo-2-2,3-d ih yd r ofu r yl}a cet a t e
(2Z). Under argon, BCl3 (223 µL, 1 M in CH2Cl2) was added
slowly to a -78 °C solution of 22Z (57.4 mg, 0.11 mmol) in
CH2Cl2 (2.0 mL), and the mixture was stirred for 2 h. Buffer
solution (pH 7.2) was then added slowly, and the reaction
mixture was diluted with Et2O. The layers were separated,
the aqueous layer was further extracted twice with CHCl3, and
the combined organics were dried over MgSO4. Purification
by silica gel column chromatography gave 2Z (42.1 mg, 89%).
IR (neat) 3432 (OH), 3022 (CH), 2959 (CH), 2929 (CH), 2857
(CH), 1749 (CdO), 1665 (CdC) cm-1 1H NMR (250 MHz,
;
CDCl3) δ 0.92-0.99 (m, 15 H, CdCHCH2CH(CH(CH3)2)2 and
CH3(CH2)6CH2CH2OC(O)CH2C), 1.14-1.23 (m, 1 H, CdCHCH2-
CH(CH(CH3)2)2), 1.27-1.45 (m, 12 H, CH3(CH2)6CH2CH2OC-
(O)CH2C), 1.61-1.75 (m, 2 H, CH3(CH2)6CH2CH2OC(O)CH2C),
1.76-1.94 (m, 2 H, CdCHCH2CH(CH(CH3)2)2), 2.37 (br s, 1
H, HOCH2C), 2.74-2.94 (m, 4 H, CdCHCH2CH(CH(CH3)2)2
and lactone-CH2), 2.99-3.00 (m, 2H, CH3(CH2)6CH2CH2OC-
(O)CH2C), 3.77 (AB quartet, J ) 12.1 Hz, 2 H, HOCH2C), 4.14
(t, J ) 6.7 Hz, 2 H, CH3(CH2)6CH2CH2OC(O)CH2C), 6.31 (tt,
J ) 7.4, 2.2 Hz, 1 H, CdCHCH2CH(CH(CH3)2)2); 13C NMR
(62.9 MHz, CDCl3) δ 14.03, 19.34 19.39, 21.55, 21.60, 22.59,
25.78, 26.25, 28.40, 29.14, 29.26, 29.30, 29.38, 31.76, 35.48,
40.39, 51.09, 65.13, 67.00, 81.88, 123.01, 147.34, 168.57,
169.38; FAB-MS (m/z, relative intensity) 425 (MH+, 100). Anal.
(C25H44O5) C, H.
(E)-2-{4-[4-Meth yl-3-(m eth yleth yl)p en tylid en e]-5-oxo-
2-[(p h e n ylm e t h oxy)m e t h yl]-2-2,3-d ih yd r ofu r yl}e t h yl
Non a n oa te (23E). According to the literature,20 nonanoyl
chloride (166 mg, 0.94 mmol) was added to a 0 °C stirring
solution of 19E (87.7 mg, 0.23 mmol) and pyridine (76 µL, 0.94
mmol) in CH2Cl2 (5 mL) under argon. The reaction was
monitored by TLC, and then the mixture was concentrated in
vacuo. Purification by silica gel column chromatography gave
23E (54 mg, 45%). IR (neat) 3026 (CH), 2959 (CH), 2929 (CH),
2858 (CH), 1747 (CdO), 1676 (CdC) cm-1; 1H NMR (250 MHz,
CDCl3) δ 0.90-0.99 (m, 15 H, CdCHCH2CH(CH(CH3)2)2 and
CH3(CH2)5CH2CH2C(O)), 1.20-1.48 (m, 11 H, CdCHCH2-
CH(CH(CH3)2)2 and CH3(CH2)5CH2CH2C(O)), 1.60-1.77 (m, 2
H, CH3(CH2)5CH2CH2C(O)), 1.78-1.93 (m, 1 H, CdCHCH2-
CH(CH(CH3)2)2), 2.05-2.25 (m, 3 H, CdCHCH2CH(CH(CH3)2)2
and OCH2CH2C), 2.30-2.37 (m, 2 H, CdCHCH2CH(CH-
(CH3)2)2), 2.43 (t, J ) 7.6 Hz, 2 H, CH3(CH2)5CH2CH2C(O)),
2.86 (br AB quartet, J ) 17.0 Hz, 2 H lactone-CH2), 3.56 (s, 2
H, PhCH2OCH2C), 4.28 (t, J ) 6.8 Hz, 2 H, OCH2CH2C), 4.63
(s, 2 H, PhCH2OCH2C), 6.80-6.89 (m, 1 H, CdCHCH2CH(CH-
(CH3)2)2), 7.34-7.43 (m, 5 H, PhCH2OCH2C); 13C NMR (62.9
MHz, CDCl3) δ 14.03, 19.24, 19.32, 21.49, 21.57, 22.57, 24.80,
28.57, 29.05, 29.07, 29.11, 29.14, 31.73, 33.29, 34.20, 35.82,
50.19, 59.33, 73.55, 74.02, 82.69, 125.45, 127.44, 127.66,
128.28, 137.38, 142.70, 169.98, 173.39; FAB-MS (m/z, relative
intensity) 515 (MH+, 22), 91 (100). Anal. (C32H50O5‚0.2H2O)
C, H.
(Z)-Non yl 2-{4-[4-Meth yl-3-(m eth yleth yl)p en tylid en e]-
5-oxo-2-[(p h en ylm et h oxy)m et h yl]-2-2,3-d ih yd r ofu r yl}-
a ceta te (22Z). Under argon, 1-nonanol (239 µL), DMAP (3.35
mg), and DCC (283 mg) were added to a 0 °C solution of 21Z
(0.274 mmol) in pyridine (0.6 mL), and the mixture was stirred
overnight warming to room temperature. The crude reaction
mixture was filtered through Celite and concentrated. Puri-
fication by silica gel column chromatography gave 22Z (79 mg,
56% for three steps from 19Z) as a colorless oil. IR (neat) 3024
(CH), 2959 (CH), 2929 (CH), 2858 (CH), 1750 (CdO), 1666
1
(CdC) cm-1; H NMR (250 MHz, CDCl3) δ 0.92-1.00 (m, 15
H, CdCHCH2CH(CH(CH3)2)2 and CH3(CH2)6CH2CH2OC(O)-
CH2C), 1.12-1.24 (m, 1 H, CdCHCH2CH(CH(CH3)2)2), 1.30-
1.45 (br s, 14 H, CH3(CH2)6CH2CH2OC(O)CH2C), 1.60-1.75
(m, 2 H, CdCHCH2CH(CH(CH3)2)2), 1.77-1.94 (m, 2 H,
CdCHCH2CH(CH(CH3)2)2), 2.63-3.17 (m, 4 H, CH3(CH2)6CH2-
CH2OC(O)CH2C and lactone-CH2), 3.64 (s, 2 H, PhCH2-
OCH2C), 4.12 (t, J ) 6.7 Hz, 2 H, CH3(CH2)6CH2CH2OC(O)-
CH2C), 4.64 (s, 2 H, PhCH2OCH2C), 6.22-6.31 (m, 1 H,
CdCHCH2CH(CH(CH3)2)2), 7.35-7.41 (m, 5 H, PhCH2OCH2C);
13C NMR (62.9 MHz, CDCl3) δ 14.06, 19.34 19.42, 21.56, 21.64,
22.61, 25.81, 26.16, 28.41, 29.17, 29.27, 29.31, 29.40, 31.78,
36.25, 40.89, 51.11, 64.94, 73.50, 74.00, 80.92, 123.30, 127.42,
127.59, 128.24, 137.51, 146.40, 168.58, 169.19; FAB-MS (m/z,
relative intensity) 515 (MH+, 29), 91 (100). Anal. (C32H50O5)
C, H.
(E)-Non yl 2-{2-(Hydr oxym eth yl)-4-[4-m eth yl-3-(m eth yl-
e t h yl)p e n t ylid e n e ]-5-oxo-2-2,3-d ih yd r ofu r yl}a ce t a t e
(2E). Under argon, BCl3 (132 µL, 1 M in CH2Cl2) was added
slowly to a -78 °C solution of 22E (34.0 mg, 0.07 mmol) in
CH2Cl2 (1.3 mL), and the mixture was stirred for 2 h. Buffer
solution (pH 7.2) was then added slowly, and the reaction
mixture was diluted with Et2O. The layers were separated,
the aqueous layer was further extracted twice with CHCl3, and
the combined organics were dried over MgSO4. Purification
by silica gel column chromatography gave 2E (26.2 mg, 93%).
IR (neat) 3422 (OH), 3021 (CH), 2960 (CH), 2929 (CH), 2857
(Z)-2-{4-[4-Meth yl-3-(m eth yleth yl)p en tylid en e]-5-oxo-
2-[(p h e n ylm e t h oxy)m e t h yl]-2-2,3-d ih yd r ofu r yl}e t h yl
Non a n oa te (23Z). According to the literature,20 nonanoyl
chloride (141 mg, 0.80 mmol) was added to a 0 °C stirring

652 J ournal of Medicinal Chemistry, 2004, Vol. 47, No. 3
Tamamura et al.
solution of 19Z (75 mg, 0.20 mmol) and pyridine (65 µL, 0.80
mmol) in CH2Cl2 (4 mL) under argon. The reaction was
monitored by TLC, and then the mixture was concentrated in
vacuo. Purification by silica gel column chromatography gave
23Z (76 mg, 74%). IR (neat) 3021 (CH), 2959 (CH), 2929 (CH),
2859 (CH), 1745 (CdO), 1666 (CdC) cm-1; 1H NMR (250 MHz,
CDCl3) δ 0.91-1.00 (m, 15 H, CdCHCH2CH(CH(CH3)2)2 and
CH3(CH2)5CH2CH2C(O)), 1.14-1.19 (m, 1 H, CdCHCH2CH(CH-
(CH3)2)2), 1.35 (br s, 10 H, CH3(CH2)5CH2CH2C(O)), 1.57-1.75
(m, 2 H, CH3(CH2)5CH2CH2C(O)), 1.76-1.94 (m, 2 H, CdCH-
CH2CH(CH(CH3)2)2), 2.0-2.25 (m, 2 H, OCH2CH2C), 2.30-2.36
(m, 2 H, CH3(CH2)5CH2CH2C(O)), 2.77-3.05 (m, 4 H, lactone-
CH2 and CdCHCH2CH(CH(CH3)2)2), 3.54 (s, 2 H, PhCH2-
OCH2C), 4.24-4.30 (m, 2 H, OCH2CH2C), 4.62 (s, 2 H,
PhCH2OCH2C), 6.29-6.40 (m, 1 H, CdCHCH2CH(CH(CH3)2)2),
7.36-7.41 (m, 5 H, PhCH2OCH2C); 13C NMR (62.9 MHz,
CDCl3) δ 14.03, 19.34, 19.38, 21.57, 21.60, 22.57, 24.79, 26.17,
29.04, 29.06, 29.14, 29.29, 31.73, 34.19, 35.61, 36.72, 51.12,
59.38, 73.50, 73.86, 82.02, 123.46, 127.44, 127.64, 128.27,
137.45, 146.34, 168.80, 173.38; FAB-MS (m/z, relative inten-
sity) 515 (MH+, 23), 91 (100). Anal. (C32H50O5‚0.3H2O) C, H.
0.62 mmol) were added to a solution of 14 (330 mg, 1.24 mmol)
in DMSO (2.5 mL), and the mixture was stirred overnight.
After 16 h, the reaction was quenched with portions of oxalic
acid (447 mg, 4.96 mmol) and the mixture was diluted with
Et2O (20 mL) and cooled to -78 °C. The resulting mixture was
filtered and washed with cold Et2O to remove excess DCU.
The organic solution was then washed with water and brine,
dried over MgSO4, and concentrated to give crude 24, which
was used directly without further purification.
(5-Meth oxy-2-pr op-2-en yloxolan -2-yl)(ph en ylm eth oxy)-
m eth a n e (25). According to the literature,29 MePPh3Br (886
mg, 2.48 mmol) was added to potassium tert-butoxide (2.48
mL, 2.48 mmol, 1.0 M in THF). After 30 min, the mixture was
added slowly to a solution of 24 (1.24 mmol) in THF (3.1 mL),
and the resulting mixture was stirred overnight. The reaction
was quenched with AcOH (0.2 mL), filtered and concentrated.
Purification by silica gel column chromatography (EtOAc/
hexane) gave an anomeric mixture of 25 (150 mg, 46%). When
feasible, the anomers were separated for characterization.
On e An om er : IR (neat) 3010 (CH), 2913 (CH), 2860 (CH),
1
1640 (CdC), 1098 (CO), 1037 (CO) cm-1; H NMR (250 MHz,
(E)-2-{2-(Hyd r oxym eth yl)-4-[4-m eth yl-3-(m eth yleth yl)-
p en tylid en e]-5-oxo-2-2,3-d ih yd r ofu r yl}eth yl Non a n oa te
(3E). Under argon, a solution of BCl3 (0.21 mL, 0.21 mmol, 1
M in CH2Cl2) was added slowly to a -78 °C solution of 23E
(54 mg, 0.11 mmol) in CH2Cl2 (2 mL). The reaction was
monitored by TLC and quenched slowly at -78 °C with pH
7.2 buffer solution, and the mixture was then diluted with
Et2O. The organic layer was washed with pH 7.2 buffer, dried,
and concentrated. Purification by chromatography gave 3E (33
mg, 73%). IR (neat) 3595 (OH), 3432 (OH), 3023 (CH), 2960
CDCl3) δ 1.84-2.13 (m, 4 H, lactone-CH2CH2), 2.45-2.49 (m,
2 H, CH2dCHCH2C), 3.38 (s, 3 H, >CHOCH3), 3.51 (AB
quartet, J ) 9.3 Hz, 2 H, PhCH2OCH2C), 4.61-4.72 (m, 2 H,
PhCH2OCH2C), 5.07-5.21 (m, 3 H, >CHOCH3 and CH2dCH-
CH2C), 5.82-5.93 (m, 1 H, CH2dCHCH2C), 7.35-7.44 (m, 5
H, PhCH2OCH2C); 13C NMR (62.9 MHz, CDCl3) δ 29.98, 32.38,
41.34, 54.25, 73.35, 76.54, 85.70, 105.59, 117.76, 127.33,
127.49, 128.13, 134.02, 138.36; FAB-MS (m/z, relative inten-
sity) 231 (MH+ - CH3OH, 51), 91 (100). Anal. (C16H22O3) C,
H.
(CH), 2930 (CH), 2872 (CH), 1746 (CdO), 1674 (CdC) cm-1
;
3-{5-Meth oxy-2-[p h en ylm eth oxy)m eth yl]oxola n -2-yl}-
p r op a n -1-ol (26). According to the literature,29 a solution of
BH3‚SMe2 in THF (482 µL, 0.96 mmol, 2 M solution) was added
dropwise to a -78 °C stirring solution of 25 (127 mg, 0.48
mmol) in THF (1.5 mL). The reaction mixture was slowly
allowed to reach 0 °C, and after 17 h, it was treated with
NaBO3‚H2O (148 mg, 0.96 mmol) at 0 °C. The reaction mixture
then stirred for 2 h warming to room temperature and was
diluted with EtOAc. The organic layer was washed with water
and brine, dried over MgSO4, and concentrated. Purification
by silica gel column chromatography (EtOAc/hexane) gave an
anomeric mixture of 26 (89 mg, 66%). IR (neat) 3446 (OH),
3012 (CH), 2952 (CH), 1098 (CO), 1037 (CO) cm-1; FAB-MS
(m/z, relative intensity) 249 (MH+ - CH3OH, 27), 91 (100).
Anal. (C16H24O4) C, H.
1H NMR (250 MHz, CDCl3) δ 0.92-1.00 (m, 15 H, CdCHCH2-
CH(CH(CH3)2)2 and CH3(CH2)5CH2CH2C(O)), 1.25-1.35 (m, 11
H, CH3(CH2)5CH2CH2C(O) and CdCHCH2CH(CH(CH3)2)2),
1.62-1.71 (m, 2 H, CH3(CH2)5CH2CH2C(O)), 1.80-1.93 (m, 2
H, CdCHCH2CH(CH(CH3)2)2), 2.05-2.26 (m, 5 H, OCH2CH2C,
CdCHCH2CH(CH(CH3)2)2 and HOCH2C), 2.36 (t, J ) 7.6 Hz,
2 H, CH3(CH2)5CH2CH2C(O)), 2.86 (br AB quartet, J ) 17.3
Hz, 2 H lactone-CH2), 3.72 (AB quartet, J ) 12.1 Hz, 2 H,
HOCH2C), 4.29 (t, J ) 6.6 Hz, 2 H, OCH2CH2C), 6.84-6.93
(m, 1 H, CdCHCH2CH(CH(CH3)2)2); 13C NMR (62.9 MHz,
CDCl3) δ 14.03, 19.27, 19.30, 21.55, 22.57, 24.79, 28.65, 29.04,
29.07, 29.14, 29.17, 31.73, 32.35, 34.19, 35.22, 50.24, 59.29,
67.09, 83.73, 125.21, 143.56, 169.93, 173.37; FAB-MS (m/z,
relative intensity) 425 (MH+, 55), 57 (100). Anal. (C25H44O5)
C, H.
1-[(4-Meth oxyp h en yl)m eth oxy]-3-{5-m eth oxy-2-[(p h en -
ylm eth oxy)m eth yl]oxola n -2-yl}p r op a n e (27). According to
the literature,33 a solution of 26 (79 mg, 0.28 mmol) in THF (3
mL) was added dropwise to a 0 °C slurry of NaH (23 mg, 0.56
mmol, 60% in oil) in THF (2 mL), followed by addition of
p-methoxybenzyl chloride (42 µL, 0.31 mmol) and Bu4NI (10
mg). The reaction mixture was then heated to reflux and
monitored by TLC. Upon completion, the reaction was cooled
to 0 °C and quenched with methanol (500 µL), followed by
saturated aqueous NH4Cl (10 mL). The aqueous solution was
then extracted with CH2Cl2, dried over MgSO4, and concen-
trated. Purification by silica gel column chromatography
(hexanes/EtOAc) gave an anomeric mixture of 27 (90 mg, 80%).
IR (neat) 3015 (CH), 2954 (CH), 2861 (CH), 1097 (CO), 1036
(CO) cm-1; FAB-MS (m/z, relative intensity) 369 (MH+ - CH3-
OH, 9), 121 (100). Anal. (C24H32O5) C, H.
(Z)-2-{2-(Hyd r oxym eth yl)-4-[4-m eth yl-3-(m eth yleth yl)-
p en tylid en e]-5-oxo-2-2,3-d ih yd r ofu r yl}eth yl Non a n oa te
(3Z). Under argon, a solution of BCl3 (0.30 mL, 0.30 mmol, 1
M in CH2Cl2) was added slowly to a -78 °C solution of 23Z
(76 mg, 0.15 mmol) in CH2Cl2 (3 mL). The reaction was
monitored by TLC and quenched slowly at -78 °C with pH
7.2 buffer solution, and then the mixture was diluted with
Et2O. The organic layer was washed with pH 7.2 buffer, dried,
and concentrated. Purification by chromatography gave 3Z (42
mg, 66%). IR (neat) 3594 (OH), 3432 (OH), 3025 (CH), 2959
(CH), 2929 (CH), 2858 (CH), 1744 (CdO), 1664 (CdC) cm-1
;
1H NMR (250 MHz, CDCl3) δ 0.92-0.99 (m, 15 H, CdCHCH2-
CH(CH(CH3)2)2 and CH3(CH2)5CH2CH2C(O)), 1.13-1.22 (m, 1
H, CdCHCH2CH(CH(CH3)2)2), 1.33-1.35 (m, 10 H, CH3(CH2)5-
CH2CH2C(O)), 1.62-1.73 (m, 2 H, CH3(CH2)5CH2CH2C(O)),
1.76-1.94 (m, 2 H, CdCHCH2CH(CH(CH3)2)2), 2.02-2.22 (m,
3 H, OCH2CH2C and HOCH2C), 2.33-2.38 (m, 2 H, CH3(CH2)5-
CH2CH2C(O)), 2.76-3.03 (m, 4 H, CdCHCH2CH(CH(CH3)2)2
and lactone-CH2), 3.69 (AB quartet, J ) 12.2 Hz, 2 H,
HOCH2C), 4.28 (t, J ) 6.6 Hz, 2 H, OCH2CH2C), 6.28-6.34
(m, 1 H, CdCHCH2CH(CH(CH3)2)2); 13C NMR (62.9 MHz,
CDCl3) δ 14.01, 19.36, 21.57, 22.55, 24.78, 26.26, 29.03, 29.05,
29.12, 29.29, 31.72, 34.19, 34.99, 35.70, 51.12, 59.34, 66.86,
83.10, 123.24, 147.21, 168.82, 173.39; FAB-MS (m/z, relative
intensity) 425 (MH+, 46), 57 (100). Anal. (C25H44O5) C, H.
5-{3-[(4-Meth oxyp h en yl)m eth oxy]p r op yl}-5-[(p h en yl-
m eth oxy)m eth yl]oxola n -2-ol{5-m eth oxy-2-[(p h en ylm eth -
oxy)m eth yl]oxola n -2-yl}p r op a n e (28). According to the
literature,33 HCl (1.4 mL, 2 N) was added to a solution of 27
(389 mg, 0.97 mmol) in THF (18.3 mL) and H2O (7.03 mL) at
0 °C. After the mixture was allowed to warm to room
temperature over a 5 h period, additional HCl (1.5 mL) was
added and the reaction mixture was stirred overnight. The
reaction was quenched with solid NaHCO3 followed by H2O.
The aqueous solution was extracted with CH2Cl2, dried over
MgSO4, and concentrated. Silica gel column chromatography
2-{5-Meth oxy-2-[(p h en ylm eth oxy)m eth yl]oxola n -2-yl}-
eth a n a l (24). DCC (1.02 g, 4.96 mmol) and DCAA (51.1 µL,

Analogues of Diacylglycerol
J ournal of Medicinal Chemistry, 2004, Vol. 47, No. 3 653
(hexane/EtOAc) gave an anomeric mixture of 28 (173 mg, 46%),
which was used without further purification.
OC6H4CH2O), 4.49 (s, 2 H, CH3OC6H4CH2O), 4.62 (s, 2 H,
PhCH2OCH2C), 6.16-6.26 (m, 1 H, CdCHCH2CH(CH(CH3)2)2),
6.95 (d, J ) 8.5 Hz, 2 H, CH3OC6H4CH2O), 7.30-7.41 (m, 7
H, PhCH2OCH2C and CH3OC6H4CH2O); 13C NMR (62.9 MHz,
CDCl3) δ 19.37, 21.59, 23.38, 26.13, 29.29, 33.68, 36.31, 51.15,
55.19, 69.65, 72.43, 73.46, 73.95, 83.37, 113.63, 124.10, 127.42,
127.54, 128.23, 129.06, 130.43, 137.69, 145.80, 158.97, 169.27;
FAB-MS (m/z, relative intensity) 507.5 (MH+ - H2, 3), 121
(100). Anal. (C32H44O5) C, H.
5-{3-[(4-Meth oxyp h en yl)m eth oxy]p r op yl}-5-[(p h en yl-
m eth oxy)m eth yl]-3,4,5-tr ih yd r ofu r a n -2-on e (29). Accord-
ing to the literature,34 solid TPAP (7.6 mg, 0.022 mmol, 5 mol
%) was added in one portion to a stirring mixture of 28 (167
mg, 0.43 mmol), NMO (75.7 mg, 0.65 mmol), and powdered 4
Å molecular sieves (190 mg heat-dried under vacuum) in CH2-
Cl2/CH3CN (0.8 mL, 10% CH3CN in CH2Cl2) at 0 °C under
argon. The reaction mixture was stirred overnight and allowed
to reach room temperature. The crude reaction mixture was
then concentrated in vacuo, filtered through Celite (eluting
with CH2Cl2) followed by filtration through silica gel (eluting
with EtOAc), and concentrated to give 29 (147 mg, 89%). IR
(E)-5-(3-Hyd r oxyp r op yl)-3-[4-m eth yl-3-(m eth yleth yl)-
p en tylid en e]-5-[(p h en ylm eth oxy)m eth yl]-4,5-d ih yd r ofu -
r a n -2-on e (31E). According to the literature procedure,32 DDQ
(31 mg, 0.14 mmol) was added to a stirring solution of 30E
(47 mg, 0.09 mmol) in CH2Cl2 (0.64 mL) and H2O (0.13 mL).
After 2.5 h, the mixture was filtered through Celite rinsing
with CH2Cl2. The organic phase was washed sequentially with
saturated aqueous NaHCO3 and brine, dried, and concen-
trated. Purification by silica gel column chromatography gave
31E (31 mg, 88%). IR (neat) 3464 (OH), 3021 (CH), 2959 (CH),
2928 (CH), 2856 (CH), 1746 (CdO), 1675 (CdC) cm-1; 1H NMR
(250 MHz, CDCl3) δ 0.91, 0.92, 0.97, and 0.98 (d, J ) 6.8 Hz,
3 H, CdCHCH2CH(CH(CH3)2)2), 1.21-1.40 (m, 1 H, CdCHCH2-
CH(CH(CH3)2)2), 1.60-2.02 (m, 6 H, CdCHCH2CH(CH(CH3)2)2
and HOCH2CH2CH2C), 2.12-2.23 (m, 2 H, CdCHCH2CH(CH-
(CH3)2)2), 2.80 (br AB quartet, J ) 17.0 Hz, 2 H, lactone-CH2),
3.56 (s, 2 H, PhCH2OCH2C), 3.73 (t, J ) 6.2 Hz, 2 H, CHOCH2-
CH2CH2C), 4.62 (AB quartet, J ) 12.2 Hz, 2 H, PhCH2OCH2C),
6.79-6.88 (m, 1 H, CdCHCH2CH(CH(CH3)2)2), 7.31-7.48 (m,
5 H, PhCH2OCH2C); 13C NMR (62.9 MHz, CDCl3) δ 19.25,
19.32, 21.51, 21.58, 26.07, 28.55, 29.12, 29.15, 33.06, 33.45,
50.19, 62.49, 73.50, 73.98, 84.04, 125.96, 127.45, 127.61,
128.26, 137.53, 142.39, 170.42; FAB-MS (m/z, relative inten-
sity) 389 (MH+, 20), 91 (100). Anal. Calcd for C24H36O4: C,
74.19; H, 9.34. Found: C, 74.17; H, 10.11.
(Z)-5-(3-Hyd r oxyp r op yl)-3-[4-m eth yl-3-(m eth yleth yl)-
p en tylid en e]-5-[(p h en ylm eth oxy)m eth yl]-4,5-d ih yd r ofu -
r a n -2-on e (31Z). According to the literature procedure,32 DDQ
(33 mg, 0.14 mmol) was added to a stirring solution of 30Z
(49 mg, 0.10 mmol) in CH2Cl2 (0.67 mL) and H2O (0.13 mL).
After 2.5 h, the mixture was filtered through Celite rinsing
with CH2Cl2. The organic phase was washed sequentially with
saturated aqueous NaHCO3 and brine, dried, and concen-
trated. Purification by silica gel column chromatography gave
31Z (22 mg, 58%) as an oil. IR (neat) 3021 (CH), 2957 (CH),
2929 (CH), 2856 (CH), 1746 (CdO), 1666 (CdC) cm-1; 1H NMR
(250 MHz, CDCl3) δ 0.93, 0.94, 0.97, and 0.98 (d, J ) 6.8 Hz,
3 H, CdCHCH2CH(CH(CH3)2)2), 1.10-1.23 (m, 1 H, CdCHCH2-
CH(CH(CH3)2)2), 1.55-1.95 (m, 6 H, CdCHCH2CH(CH(CH3)2)2
and HOCH2CH2CH2C), 2.66-3.06 (m, 4 H, CdCHCH2CH(CH-
(CH3)2)2 and lactone-CH2), 3.54 (s, 2 H, PhCH2OCH2C), 3.73
(t, J ) 6.2 Hz, 2 H, HOCH2CH2CH2C), 4.63 (app s, 2 H, PhCH2-
OCH2C), 6.20-6.28 (m, 1 H, CdCHCH2CH(CH(CH3)2)2), 7.36-
7.42 (m, 5 H, PhCH2OCH2C); 13C NMR (62.9 MHz, CDCl3) δ
19.37, 21.60, 21.61, 26.16, 29.30, 29.64, 33.21, 36.50, 51.15,
62.59, 73.48, 73.82, 83.26, 123.93, 127.47, 127.60, 128.26,
137.60, 146.05, 169.22; FAB-MS (m/z, relative intensity) 389
(MH+, 14), 91 (100). Anal. (C24H36O4) C, H.
(neat) 3020 (CH), 2931 (CH), 2861 (CH), 1766 (CdO) cm-1
;
1H NMR (250 MHz, CDCl3) δ 1.71-1.89 (m, 3 H, OCH2CH2-
CHHC), 1.99-2.12 (m, 2 H, OCH2CH2CHHC and lactone-
CHHCH2), 2.30-2.39 (m, 1 H, lactone-CHHCH2), 2.51-2.62
(m, 1 H, lactone-CH2CHH), 2.72-2.84 (m, 1 H, lactone-CH2-
CHH), 3.50-3.64 (m, 4 H, OCH2CH2CH2C and PhCH2OCH2C),
3.87 (s, 3 H, CH3OC6H4CH2O), 4.50 (s, 2 H, CH3OC6H4CH2O),
4.61 (br AB quartet, J ) 12.0 Hz, 2 H, PhCH2OCH2C), 6.94-
6.98 (d, J ) 8.8 Hz, 2 H, CH3OC6H4CH2O), 7.31-7.45 (m, 7
H, PhCH2OCH2C and CH3OC6H4CH2O); 13C NMR (62.9 MHz,
CDCl3) δ 23.64, 28.78, 29.52, 33.76, 55.18, 69.56, 72.48, 73.51,
74.66, 87.23, 113.67, 127.40, 127.64, 128.32, 129.09, 130.32,
137.61, 159.02, 177.02; FAB-MS (m/z, relative intensity) 385
(MH+, 7), 121 (100). Anal. (C23H28O5) C, H.
5-{3-[(4-Meth oxyp h en yl)m eth oxy]p r op yl}-3-[4-m eth yl-
3-(m eth yleth yl)pen tyliden e]-5-[(ph en ylm eth oxy)m eth yl]-
4,5-d ih yd r ofu r a n -2-on e (30E/Z). Under argon, a -78 °C
solution of 29 (142 mg, 0.37 mmol) in THF (1.4 mL) was
treated dropwise with LDA (0.26 mL, 0.52 mmol, 2 M solution
in heptane/THF/ethylbenzene) and stirred for 2 h. A solution
of 4-methyl-3-(methylethyl)pentanal (95 mg, 0.67 mmol) in
THF (0.3 mL) was added dropwise maintaining the same
temperature and the reaction was monitored by TLC. The
reaction mixture was quenched with saturated aqueous NH4-
Cl, and then warmed to room temperature. The aqueous layer
was extracted with Et2O, washed with water, dried, and
concentrated. The residue was then taken up in CH2Cl2 (3.5
mL) under argon, cooled to 0 °C, treated sequentially with
MsCl (0.57 µL, 0.74 mmol) and Et3N (206 µL, 1.48 mmol), and
stirred at 0 °C for 30 min and then at room temperature for 2
h. The reaction was then cooled again to 0 °C, DBU (277 µL,
1.85 mmol) was added, and the resultant solution was stirred
at room temperature for 3 h. The crude reaction mixture was
filtered through a pad of silica gel and concentrated. Purifica-
tion by silica gel column chromatography gave 30E (58 mg,
31%) and 30Z (58 mg, 31%).
30E: IR (neat) 3020 (CH), 2959 (CH), 2930 (CH), 2870 (CH),
1
1745 (CdO), 1675 (CdC) cm-1; H NMR (250 MHz, CDCl3) δ
0.90-0.99 (m, 12 H, CdCHCH2CH(CH(CH3)2)2), 1.25-1.42 (m,
1 H, CdCHCH2CH(CH(CH3)2)2), 1.64-1.89 (m, 6 H, CdCHCH2-
CH(CH(CH3)2)2)2 and OCH2CH2CH2C), 2.13-2.19 (m, 2 H,
CdCHCH2CH(CH(CH3)2)2), 2.80 (br AB quartet, J ) 17.0 Hz,
2 H, lactone-CH2), 3.50-3.55 (m, 2 H, OCH2CH2CH2C), 3.55
(s, 2 H, PhCH2OCH2C), 3.89 (s, 3 H, CH3OC6H4CH2O), 4.49
(s, 2 H, CH3OC6H4CH2O), 4.56-4.70 (m, 2 H, PhCH2OCH2C),
6.78-6.86 (m, 1 H, CdCHCH2CH(CH(CH3)2)2), 6.95 (d, J )
8.8 Hz, 2 H, CH3OC6H4CH2O), 7.34-4.41 (m, 7 H, PhCH2-
OCH2C and CH3OC6H4CH2O); 13C NMR (62.9 MHz, CDCl3) δ
19.25, 19.32, 21.50, 21.57, 23.31, 28.51, 29.12, 29.14, 32.90,
33.92, 50.18, 55.19, 69.57, 72.43, 73.49, 74.09, 84.11, 113.64,
126.12, 127.40, 127.56, 128.23, 129.06, 130.32, 137.61, 142.11,
158.99, 170.42; FAB-MS (m/z, relative intensity) 509.5 (MH+,
3), 121 (100). Anal. (C32H44O5‚0.3H2O) C, H.
(E)-3-{4-[4-Meth yl-3-(m eth yleth yl)p en tylid en e]-5-oxo-
2-[(ph en ylm eth oxy)m eth yl]-2-2,3-dih ydr ofu r yl}pr opyl Oc-
ta n oa te (32E). According to the literature,20 octanoyl chloride
(44 µL, 0.26 mmol) was added to a 0 °C stirring solution of
31E (24.9 mg, 0.06 mmol) and pyridine (21 µL, 0.26 mmol) in
CH2Cl2 (1.5 mL) under argon. The reaction was monitored by
TLC, and upon completion, the mixture was concentrated in
vacuo. Purification by silica gel column chromatography gave
32E (28 mg, 86%). IR (neat) 3021 (CH), 2959 (CH), 2930 (CH),
2859 (CH), 1744 (CdO), 1675 (CdC) cm-1; 1H NMR (250 MHz,
CDCl3) δ 0.90-1.00 (m, 3 H, CH3(CH2)4CH2CH2C(O)), 0.91,
0.92, 0.97, and 0.98 (d, J ) 6.8 Hz, 3 H, CdCHCH2CH(CH-
(CH3)2)2), 1.24-1.45 (m, 9 H, CdCHCH2CH(CH(CH3)2)2 and
CH3(CH2)4CH2CH2C(O)), 1.60-1.98 (m, 8 H, CdCHCH2CH-
(CH(CH3)2)2, OCH2CH2CH2C and CH3(CH2)4CH2CH2C(O)),
2.15-2.20 (m, 2 H, CdCHCH2CH(CH(CH3)2)2), 2.36 (t, J ) 7.6
Hz, 2 H, CH3(CH2)4CH2CH2C(O)), 2.80 (br AB quartet, J )
30Z: IR (neat) 3015 (CH), 2959 (CH), 2931 (CH), 2870 (CH),
1
1745 (CdO), 1665 (CdC) cm-1; H NMR (250 MHz, CDCl3) δ
0.92-1.0 (m, 12 H, CdCHCH2CH(CH(CH3)2)2), 1.09-1.23 (m,
1 H CdCHCH2CH(CH(CH3)2)2), 1.65-1.94 (m, 6 H, CdCHCH2-
CH(CH(CH3)2)2 and OCH2CH2CH2C), 2.62-3.03 (m, 4 H,
CdCHCH2CH(CH(CH3)2)2 and lactone-CH2), 3.49-3.54 (m, 4
H, PhCH2OCH2C and OCH2CH2CH2C), 3.89 (s, 3 H, CH3-

654 J ournal of Medicinal Chemistry, 2004, Vol. 47, No. 3
Tamamura et al.
17.0 Hz, 2 H, lactone-CH2), 3.55 (AB quartet, J ) 10.0 Hz, 2
H, PhCH2OCH2C), 4.13 (t, J ) 6.0 Hz, 2 H, OCH2CH2CH2C),
4.62 (AB quartet, J ) 12.1 Hz, 2 H, PhCH2OCH2C), 6.80-
6.89 (m, 1 H, CdCHCH2CH(CH(CH3)2)2), 7.36-7.45 (m, 5 H,
PhCH2OCH2C); 13C NMR (62.9 MHz, CDCl3) δ 14.02, 19.25,
19.30, 21.51, 21.57, 22.37, 22.54, 24.90, 28.56, 28.85, 29.05,
29.12, 29.14, 31.59, 33.09, 33.65, 34.21, 50.19, 63.76, 73.51,
73.83, 83.61, 125.77, 127.45, 127.65, 128.28, 137.48, 142.53,
170.24, 173.63; FAB-MS (m/z, relative intensity) 515 (MH+,
49), 91 (100). Anal. (C32H50O5) C, H.
2857 (CH), 1739 (CdO), 1664 (CdC) cm-1; 1H NMR (250 MHz,
CDCl3) δ 0.93-1.00 (m, 15 H, CdCHCH2CH(CH(CH3)2)2 and
CH3(CH2)4CH2CH2C(O)), 1.13-1.24 (m, 1 H, CdCHCH2CH(CH-
(CH3)2)2), 1.28-1.44 (m, 8 H, CH3(CH2)4CH2CH2C(O)), 1.60-
1.95 (m, 8 H, CdCHCH2CH(CH(CH3)2)2, CH3(CH2)4CH2CH2C-
(O) and OCH2CH2CH2C), 2.02 (br s, 1 H, HOCH2C), 2.37 (t, J
) 7.6 Hz, 2 H, CH3(CH2)4CH2CH2C(O)), 2.67-3.03 (m, 4 H,
CdCHCH2CH(CH(CH3)2)2 and lactone-CH2), 3.69 (AB quartet,
J ) 12.1 Hz, 2 H, HOCH2C), 4.13-4.17 (m, 2 H, OCH2CH2-
CH2C), 6.27-6.35 (m, 1 H, CdCHCH2CH(CH(CH3)2)2); 13C
NMR (62.9 MHz, CDCl3) δ 14.01, 19.33, 19.39, 21.57, 21.60,
22.43, 22.53, 24.89, 26.27, 28.84, 29.04, 29.29, 29.32, 31.59,
32.77, 34.20, 35.12, 51.15, 63.74, 66.97, 83.98, 123.58, 147.05,
169.07, 173.64; FAB-MS (m/z, relative intensity) 425 (MH+,
48), 57 (100). Anal. Calcd for C25H44O5: C, 70.72; H, 10.44.
Found: C, 71.27; H, 10.73.
(Z)-3-{4-[4-Meth yl-3-(m eth yleth yl)p en tylid en e]-5-oxo-
2-[(ph en ylm eth oxy)m eth yl]-2-2,3-dih ydr ofu r yl}pr opyl Oc-
ta n oa te (32Z). According to the literature,20 octanoyl chloride
(38 µL, 0.20 mmol) was added to a 0 °C stirring solution of
31Z (19.2 mg, 0.05 mmol) and pyridine (16 µL, 0.20 mmol) in
CH2Cl2 (1.2 mL) under argon. The reaction was monitored by
TLC, and upon completion, the mixture was concentrated in
vacuo. Purification by silica gel column chromatography gave
32Z (23 mg, 91%). IR (neat) 3022 (CH), 2958 (CH), 2929 (CH),
2857 (CH), 1741 (CdO), 1666 (CdC) cm-1; 1H NMR (250 MHz,
CDCl3) δ 0.91-1.00 (m, 15 H, CH3(CH2)4CH2CH2C(O) and
CdCHCH2CH(CH(CH3)2)2), 1.12-1.23 (m, 1 H, CdCHCH2-
CH(CH(CH3)2)2), 1.30-1.45 (m, 10 H, OCH2CH2CH2C and CH3-
(CH2)4CH2CH2C(O)), 1.60-1.97 (m, 6 H, CdCHCH2CH-
(CH(CH3)2)2, OCH2CH2CH2C and CH3(CH2)4CH2CH2C(O)),
2.36 (t, J ) 7.5 Hz, 2 H, CH3(CH2)4CH2CH2C(O)), 2.66-3.02
(m, 4 H, lactone-CH2 and CdCHCH2CH(CH(CH3)2)2), 3.52 (s,
2 H, PhCH2OCH2C), 4.13 (t, J ) 5.8 Hz, 2 H, OCH2CH2CH2C),
4.63 (app s, 2 H, PhCH2OCH2C), 6.20-6.29 (m, 1 H, CdCHCH2-
CH(CH(CH3)2)2), 7.35-7.42 (m, 5 H, PhCH2OCH2C); 13C NMR
(62.9 MHz, CDCl3) δ 14.00, 19.36, 21.60, 22.43, 22.53, 24.90,
26.18, 28.84, 29.04, 29.30, 29.31, 31.59, 33.40, 34.21, 36.49,
51.15, 63.83, 73.48, 73.69, 82.90, 123.78, 127.45, 127.62,
128.26, 137.56, 146.19, 169.07, 173.63; FAB-MS (m/z, relative
intensity) 515 (MH+, 27), 91 (100). Anal. Calcd for C32H50O5:
C, 74.67; H, 9.79. Found: C, 75.47; H, 10.06.
Refer en ces
(1) Quest, A. F. G. Regulation of protein kinase C: A tale of lipids
and proteins. Enzyme Protein 1996, 49, 231-261.
(2) Mellor, H.; Parker, P. J . The extended protein kinase C super-
family. Biochem. J . 1998, 332, 281-292.
(3) Ron, D.; Kazanietz, M. G. New insights into the regulation of
protein kinase C and novel phorbol ester receptors. FASEB J .
1999, 13, 1658-1676.
(4) Newton, A. C. Protein kinase C: Structural and spatial regula-
tion by phosphorylation, cofactors, and macromolecular interac-
tions. Chem. Rev. 2001, 101, 2353-2364.
(5) Castagna, M.; Takai, Y.; Kaibuchi, K.; Sano, K.; Kikkawa, U.;
et al. Direct activation of calcium-activated, phospholipid-
dependent protein kinase by tumor-promoting phorbol esters.
J . Biol. Chem. 1982, 257, 7847-7851.
(6) Nishizuka, Y. Studies and Perspectives of Protein-Kinase-C.
Science 1986, 233, 305-312.
(7) Newton, A. C.; Keranen, L. M. Phosphatidyl-L-Serine Is Neces-
sary for Protein-Kinase Cs High-Affinity Interaction with Dia-
cylglycerol-Containing Membranes. Biochemistry 1994, 33, 6651-
6658.
(8) Newton, A. C. Regulation of protein kinase C. Curr. Opin. Cell
Biol. 1997, 9, 161-167.
(E)-3-{2-(Hyd r oxym eth yl)-4-[4-m eth yl-3-(m eth yleth yl)-
p en tylid en e]-5-oxo-2-2,3-d ih yd r ofu r yl}p r op yl Octa n oa te
(4E). Under argon, BCl3 (94 µL, 0.094 mmol, 1 M in CH2Cl2)
was added slowly to a -78 °C solution of 32E (24.3 mg, 0.047
mmol) in CH2Cl2 (1.0 mL) and stirred for 2 h. Buffer solution
(pH 7.2) was added slowly, and the reaction mixture was
diluted with Et2O and warmed to room temperature. The
layers were separated, the aqueous layer was further extracted
with CHCl3 (2×), and the combined organics were dried over
MgSO4. Purification by silica gel column chromatography gave
4E (19 mg, 93%). IR (neat) 3596 (OH), 3023 (CH), 2960 (CH),
2829 (CH), 2872 (CH), 1742 (CdO), 1674 (CdC) cm-1; 1H NMR
(250 MHz, CDCl3) δ 0.92-1.00 (m, 15 H, CH3(CH2)4CH2-
CH2C(O) and CdCHCH2CH(CH(CH3)2)2), 1.24-1.45 (m, 9 H,
CH3(CH2)4CH2CH2C(O) and CdCHCH2CH(CH(CH3)2)2), 1.60-
1.95 (m, 8 H, CH3(CH2)4CH2CH2C(O), OCH2CH2CH2C and
CdCHCH2CH(CH(CH3)2)2), 2.08 (br s, 1 H, HOCH2C), 2.15-
2.25 (m, 2 H, CdCHCH2CH(CH(CH3)2)2), 2.36 (t, J ) 7.4 Hz,
2 H, CH3(CH2)4CH2CH2C(O)), 2.80 (br AB quartet, J ) 17.1
Hz, 2 H, lactone-CH2), 3.71 (AB quartet, J ) 12.0 Hz, 2 H,
HOCH2C), 4.13-4.17 (m, 2 H, OCH2CH2CH2C), 6.82-6.91 (m,
1 H, CdCHCH2CH(CH(CH3)2)2); 13C NMR (62.9 MHz, CDCl3)
δ 14.00, 19.27, 19.29, 21.53, 21.57, 22.35, 22.52, 24.89, 28.64,
28.84, 29.04, 29.14, 29.18, 31.58, 32.83, 33.01, 34.20, 50.23,
63.68, 67.12, 84.74, 125.62, 143.28, 170.27, 173.64; FAB-MS
(m/z, relative intensity) 425 (MH+, 66), 57 (100). Anal. (C25H44O5)
C, H.
(Z)-3-{2-(Hyd r oxym eth yl)-4-[4-m eth yl-3-(m eth yleth yl)-
p en tylid en e]-5-oxo-2-2,3-d ih yd r ofu r yl}p r op yl Octa n oa te
(4Z). Under argon, BCl3 (79 µL, 0.08 mmol, 1 M in CH2Cl2)
was added slowly to a -78 °C solution of 32Z (20.4 mg, 0.04
mmol) in CH2Cl2 (1.0 mL) and stirred for 2 h. Buffer solution
(pH 7.2) was added slowly, and the reaction mixture was
diluted with Et2O and warmed to room temperature. The
layers were separated, the aqueous layer was further extracted
with CHCl3 (2×), and the combined organics were dried over
MgSO4. Purification by silica gel column chromatography gave
4Z (16 mg, 93%). IR (neat) 3591 (OH), 2959 (CH), 2929 (CH),
(9) Nishizuka, Y. Intracellular Signaling by Hydrolysis of Phospho-
lipids and Activation of Protein-Kinase-C. Science 1992, 258,
607-614.
(10) Oancea, E.; Teruel, M. N.; Quest, A. F. G.; Meyer, T. Green
fluorescent protein (GFP)-tagged cysteine-rich domains from
protein kinase C as fluorescent indicators for diacylglycerol
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