Versatile synthesis of 1-O-(Iω-aminolauryl)-I(4,5)P2

Article abstract of DOI:10.1002/ejoc.201200726

The synthesis of a model 1-O-(Iω-aminoacyl)-IP₂ derivative, lauryl 4,5-bisphosphate 31, was realized following a versatile and high-yielding scheme. The flexible synthetic strategy used for this purpose allows the preparation of a range of othe

Full text of DOI:10.1002/ejoc.201200726

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FULL PAPER
DOI: 10.1002/ejoc.201200726
Versatile Synthesis of 1-O-(ω-Aminolauryl)-I(4,5)P2
Anastasia-Aikaterini C. Varvogli,[a] Konstantina C. Fylaktakidou,[b] Theodora Farmaki,[c]
John G. Stefanakis,[a] and Alexandros E. Koumbis*[a]
Keywords: Bioorganic chemistry / Natural products / Inositols / Phosphates / Fatty acids / Lipids
The synthesis of a model 1-O-(ω-aminoacyl)-IP2 derivative,
lauryl 4,5-bisphosphate 31, was realized following a versatile
and high-yielding scheme. The flexible synthetic strategy
used for this purpose allows the preparation of a range of
other useful IP, IP2 and IP3 derivatives. In view of the central
role played by IPs and PIPs in cellular life, the preparation of
functionalized myo-inositol derivatives of this type may facil-
itate the isolation and fluorescent localization of related pro-
teins.
PIPs are known to undergo phosphorylation at multiple
sites to generate diverse phosphoinositides, which, in turn,
regulate a series of signal-transduction pathways.[1] They
can undergo sequential and reversible phosphorylations at
the 3-, 4-, and 5-positions, catalysed by specific kinases.
Consequently, they act as important mediators of several
biochemical processes that are known to play central roles
in fundamental cellular functions, such as intracellular vesi-
cle trafficking, apoptosis, cell proliferation, and metabo-
lism.[3]
Introduction
myo-Inositol phosphates (IPs) play a key role in bio-
logical systems. They function as secondary messengers in
important cell-signalling pathways.[1] In particular, phos-
phatidylinositol (PI, 1, Figure 1) and its ring-phosphory-
lated derivatives (PIPs, 2) comprise a very important class
of membrane phospholipids.[2] Naturally occurring PIPs
can be categorized according to their inositol ring phos-
phorylation pattern and the fatty acids present on the glyc-
erol moiety.
More specifically, PI(4,5)P2 (3, Figure 1) has been impli-
cated in a variety of important cellular functions. It is hy-
drolyzed by a phosphatidylinositol-specific phospholipase
C (PLC) to generate two important secondary messenger
molecules, I(1,4,5)P3 and diacylglycerol.[4] This initiates an
important pathway for cell proliferation in which I(1,4,5)P3
interacts with membrane-bound receptors to release Ca2+
,
which is a key event in cellular signal transduction.[5] This
process mediates cellular responses to hormones, neuro-
transmitters, and other cellular signalling molecules, includ-
ing odorants and those responsible for bitter taste.[6] In ad-
dition, PI(4,5)P2 sequesters profilin to the inner surface of
the phospholipid bilayer, which facilitates the polymeriza-
tion of actin.[7] It can also be converted by agonist-stimu-
lated, receptor-mediated activation of phosphoinositide 3-
kinase (PI 3-K) to PI(3,4,5)P3, the key element in a newly
discovered intracellular signaling system.[8]
Because of their unique biological profile, several re-
search groups have designed and synthesized a number of
PIP analogues.[2,9] These modified PIPs could be helpful
probes for examining the stereospecificity and inhibition of
several inositol monophosphatase enzymes, and for explor-
ing their potential use as drugs. They mainly belong to the
families of inositol-deoxygenated analogues,[10,11] phos-
phorothioates,[12–14] and phosphonates,[13,15] but other types
of PIP analogue have also been synthesized.[16,17]
Figure 1. General structures of PI (1), PIPs (2 and 3) and structures
of known 1-O-acyl-IPs (4–6) [APB = 4-(4-aminophenyl)butyryl].
[a] Laboratory of Organic Chemistry, Department of Chemistry,
Aristotle University of Thessaloniki,
Thessaloniki 54124, Greece
Fax: +30-2310-997679
E-mail: akoumbis@chem.auth.gr
Homepage: http://www.chem.auth.gr/index.php?lang=en&st=17
[b] Department of Molecular Biology and Genetics, Democritus
University of Thrace,
68100 Alexandroupolis, Greece
[c] Institute of Applied Biosciences, Centre for Research and
Technology, Hellas-CERTH,
57001 Thermi, Greece
Supporting information for this article is available on the
WWW under http://dx.doi.org/10.1002/ejoc.201200726.
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In order to clarify the diverse biological role of PIPs, it pound could be derived from fully protected ester 8 by de-
is necessary to identify the related receptor proteins. In protection. Obviously, it would be highly desirable to realize
practice though, this is prohibited due to the lack of suf- this task in a single step, so the protecting groups (PGs)
ficient amounts of PIPs that can be obtained from natural present other than Bn should be chosen carefully. In turn,
sources. This issue was very nicely addressed by the design ester 8 leads back to advanced myo-inositol derivative 9 and
and synthesis of ω-amino-PIP analogues for the prepara- protected 12-aminolauric acid 10. The former compound is
tion of photoaffinity-labeled probes,[18,19] biotinylated expected to be derived from diol 11 by sequential phos-
probes, and others,[17,19,20] as well as affinity probes immo- phorylation and selective deprotection of the 1-position. Fi-
bilized on beads,[21] which could help in the investigation nally, 11 could be obtained from parent myo-inositol (12)
and isolation of particular receptor proteins.
by a series of regioselective protecting group manipulations
A less investigated category of PIP analogues includes and a key resolution step.
those derivatives that completely lack the phosphoglycerol
Since this synthetic plan should be widely applicable, and
tail, having replaced it by an acyl group, i.e., 1-O-acyl-IPs not only relevant to our model compound 7, the following
(Figure 1). These compounds have the advantage of being structural variations were taken into consideration: (a) the
more easily prepared, since they contain only the head- number and position of the phosphate groups, i.e., the pos-
group of a PIP. Interestingly, 1-O-stearoyl-I(3,4,5)P3 4 was sibility to construct all seven phosphorylated IPs (variables
found to completely inhibit PI 3-kinase,[22] whereas affigel- on the 3-, 4- and 5-positions); (b) the length and saturation
immobilized I(3,4,5)P3-APB 5 has shown better binding of the ω-amino fatty acid; and finally (c) the absolute ste-
selectivity to proteins than the corresponding PIP.[11,23] Ra- reochemistry, i.e., the ability to prepare the corresponding
cemic 1-O-palmitoyl-I(4,5)P2 6 has also been synthesized as enantiomeric IPs [3-O-(ω-aminoacyl)-derivatives].
a potential phospholipase C inhibitor.[24]
The key resolution step was realized early in our synthe-
In a continuation of our previous research work[25] into sis (Scheme 2). Thus, racemic 2,3:4,5-di-O-cyclohexylidene
the synthesis and biological evaluation of polyphosphoryl- myo-inositol (13), readily available on a multi-gram scale
ated inositol and carbohydrate derivatives, we wish to re- from myo-inositol,[27] was selected as the starting material
port here a versatile and high-yielding synthesis of new 1- of choice. Regioselective camphanylation of 13 with 1-(S)-
O-(ω-aminoacyl)-IP2 7 (Scheme 1). This model compound (–)-camphanyl chloride by a known procedure[28] yielded a
could potentially be used for the selective isolation of pro- 1:1 mixture of diastereoisomers 14 and 15. In order to avoid
teins (as an immobilized probe)[11,23] and/or as a fluorescent time-consuming chromatographic techniques, we examined
probe (after the attachment of a fluorescent label).[26]
the possibility of separating the two diasteroisomers after
the next step. After some experimentation, and to our de-
light, introduction of a methoxymethyl group on the free
hydroxy (C-6) furnished a 1:1 mixture of the corresponding
fully masked derivatives 16 and 17, which were easily sepa-
rated by simple fractional crystallization. The absolute con-
figuration of each diastereoisomer (16 and 17) was verified
Scheme 1. Retrosynthetic analysis (PG = protecting group).
Results and Discussion
Scheme 2. (i) Ref.[28] (ii) MOMCl, DIPEA, DMF, room temp., 72 h
(89%; 1:1 mixture of 16 and 17) (MOM = methoxymethyl; DIPEA
For the synthesis of 7, we envisaged the retrosynthesis
shown in Scheme 1. According to this plan, the target com- = diisopropylethylamine).
2
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Versatile Synthesis of 1-O-(ω-Aminolauryl)-I(4,5)P2
by their independent preparation from small quantities of present. Protecting the resulting free hydroxy groups with
pre-purified known camphanyl esters 14 and 15. This de- the same PG1 led to known tris-MOM ether 23,[29] which
cided the nature of the required protecting group PG1 was further debenzoylated to give 4,5-diol 24.[29] This com-
(Scheme 1), and thus, significantly, determined the final pound was then very cleanly phosphorylated to give 25, in
global deprotection conditions for the realization of our contrast to what has previously been reported,[29] using a
synthetic plan. Moreover, the ability to easily obtain both modified procedure. Finally, desilylation of 25 on exposure
enantiomers in pure form clearly addresses the issue of the to TBAF (tetrabutylammonium fluoride) gave alcohol 26.
future synthesis of 3-O-(ω-aminoacyl)-IPs using the same It is worth mentioning that the above described reaction
methodology.
sequence allows, after appropriate modifications (e.g., the
Having solved the issues of optical resolution and PG1 use of tin acetal chemistry), the preparation of all seven
introduction in the early stages, we proceeded with the step- possible IPs phosphorylated at the 3-, 4- and 5-positions.
wise construction of the desired phosphorylated inositol
With inositol head precursor 9 prepared, we then pro-
head-group (i.e., 9). Saponification of camphanyl ester 16 ceeded to synthesize the required protected amino acid 10,
smoothly furnished free alcohol 18 (Scheme 3). At this and for this purpose, a Cbz (benzyloxycarbonyl) group
point, for the introduction of PG3, the orthogonal TBDPS (PG2) was selected. Direct Cbz protection of 12-amino-
(tert-butyldimethylsilyl) group was chosen. Silylation oc- lauric acid to 29 is known,[30] but in our hands, it was prac-
curred in excellent yield, although it required a prolonged tically impossible to obtain the product in a form pure
reaction time. Selective removal of the more labile 4,5-cyclo- enough to use in the next step. Therefore, we adopted a
hexylidene group in 19 was achieved almost quantitatively two-step sequence with the corresponding methyl ester (i.e.,
by mild acidic methanolysis, and the resulting trans-diol 27)[31] as starting material, which involved Cbz-protection
(i.e., 20) was benzoylated under standard conditions to give to give 28 and subsequent saponification (Scheme 4). It is
ester 21. The next step required removal of the more stable obvious that any other ω-amino acid would also be suitable
2,3-cyclohexylidene ketal. A carefully monitored hydrolysis for our synthetic scheme, but the final deprotection step
protocol with aqueous trifluoroacetic acid (TFA) in di- should be compatible with the nature of fatty chain (i.e.,
chloromethane at low temperature was used to obtain cis- whether or not it is saturated).
diol 22 without cleavage of the other acid-sensitive groups
Scheme 4. (i) CbzCl, Et3N, DMAP, CH2Cl2, 0 °C to room temp.,
12 h (89%); (ii) LiOH, THF, H2O, room temp., 10 h (95%) [DMAP
= 4-(dimethylamino)pyridine].
At this point, our synthetic plan involved the coupling
reaction of the inositol head and the fatty acid tail partners.
To this end, our initial effort to promote the esterification
of 29 with 25 under simple conditions [DCC (N,NЈ-dicyclo-
hexylcarbodiimide) and DMAP in dry CH2Cl2] gave,
slowly, but very cleanly, fully protected target compound 30
(Scheme 5).
In contrast, the final deprotection step proved to be quite
challenging. Thus, we first tried to simultaneously remove
Bn and Cbz groups from 30 by Pd(OH)2 catalyzed hydro-
genolysis in tBuOH/H2O under forcing conditions (25 bar
of H2) (Scheme 5). We hoped that the acidity of the liber-
ated phosphates would be enough to cleave the MOM
ethers, either in situ or after the addition of EtSH. However,
although the Bn groups were efficiently removed in the first
2 h, the Cbz group required more than 24 h to be com-
Scheme 3. (i) LiOH, THF, H2O, room temp., 1 h (98%);
pletely cleaved, and in the meantime, we observed slow de-
(ii) TBDPSCl, imidazole, pyridine, 65 °C, 5 d (97%); (iii) AcCl,
composition in the 1H and 31P NMR spectra. Changing the
CH2Cl2, MeOH, 0 °C, 90 min (99%); (iv) BzCl, pyridine, CH2Cl2,
0 °C to room temp., 24 h (96%); (v) 90% aq. TFA, CH2Cl2, –20 °C,
2 h (94%); (vi) MOMCl, DIPEA, DMF, 75 °C, 4 d (100%);
(vii) 20% w/v MeONa in MeOH, room temp., 1 h (92%);
(viii) 1. (BnO)2PN(iPr)2, 1H-tetrazole, MeCN, room temp., 24 h;
2. mCPBA, CH2Cl2, –40 to 0 °C, 3 h (88%); (ix) TBAF, THF, room
temp., 1 h (93%) (mCPBA = meta-chloroperbenzoic acid).
solvent system to EtOH/H2O or THF/H2O did not improve
this result. We believe that self-catalyzed hydrolysis of phos-
phates and/or acid-catalyzed hydrolysis of the lauric ester
moiety occurred. Since it was impossible to perform hydro-
genolysis in the presence of a base,[32] our second approach
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A. E. Koumbis et al.
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Experimental Section
General Methods: All commercially available reagent-grade chemi-
cals were used without further purification. All solvents were puri-
fied by standard procedures before use. Dry solvents were obtained
by literature methods and stored over molecular sieves. Whenever
possible, reactions were monitored using commercially available
precoated TLC plates (layer thickness 0.25 mm) of Kieselgel 60
F254. Compounds were visualized by use of a UV lamp and/or
using a suitable stain [p-anisaldehyde ethanolic solution, phos-
phomolybdic acid (PMA), Seebach’s stain (PMA + cerium sulfate)
and warming]. Column chromatography was performed in the us-
ual way using Merck 60 (40–60 mm) silica gel. Optical rotations
were determined at room temperature with an A. Krüss P3000
Automatic Digital Polarimeter. NMR spectra were recorded with
a
300 MHz spectrometer (1H: 300 MHz, 13C: 75 MHz, 31P:
Scheme 5. (i) DCC, DMAP, CH2Cl2, room temp., 3 d (96%);
(ii) 1. TMSBr, CH2Cl2, –78 °C to room temp., 2 h; 2. MeOH, room
temp., 1 h; 3. NaHCO3, H2O, 1 h (81%).
100 MHz) in the deuterated solvent indicated. Chemical shifts are
given in parts per million and J values in Hertz using solvent or
TMS as an internal reference. Mass spectra were obtained using
the electrospray technique, positive mode (ESI-MS).
1-[1-(S)-Camphanyl]-2,3:4,5-di-O-cyclohexylidene-6-O-methoxy-
methyl-myo-inositol (16) and 3-[1-(S)-Camphanyl]-1,2:5,6-di-O-
cyclohexylidene-4-O-methoxymethyl-myo-inositol (17): A 1:1 mix-
ture of camphanyl esters 14 and 15 (880 mg, 1.7 mmol)[28] was dis-
solved in dry DMF (12 mL) under an Ar atmosphere. DIPEA
(15 mL, 85 mmol) was added followed by the dropwise addition of
MOMCl (5.2 mL, 68 mmol). The mixture was left under vigorous
stirring at room temperature for 72 h. Then it was quenched with
saturated aqueous NH4Cl (20 mL) and extracted with EtOAc(2ϫ
75 mL). The combined organic extracts were washed with brine
(2ϫ 40 mL), dried (Na2SO4), and concentrated under reduced
pressure. The residue was purified by flash column chromatography
(10Ǟ20% EtOAc in hexanes) to give a 1:1 mixture of MOM ethers
16 and 17 (855 mg, 89%). Pure diastereoisomer 16 was obtained
upon crystallization from a mixture of acetone/light petroleum spi-
rit (boiling range 35–60 °C); the other diastereomer (17) remained
in the filtrate. 16: Rf = 0.22 (25% EtOAc in hexanes). [α]D = –20.9
(c = 2, CHCl3). 1H NMR (300 MHz, CDCl3): δ = 5.19 (t, J =
5.4 Hz, 1 H, 1-H), 4.86, 4.75 (ABq, J = 6.6 Hz, 2 H, OCH2O), 4.61
(t, J = 5.3 Hz, 1 H, 2-H), 4.31 (dd, J = 8.3, 5.4 Hz, 1 H, 3-H), 4.10
(dd, J = 9.3, 5.4 Hz, 1 H, 6-H), 3.83 (dd, J = 10.4, 8.3 Hz, 1 H, 4-
H), 3.44 (dd, J = 10.4, 9.3 Hz, 1 H, 5-H), 3.42 (s, 3 H, OCH3),
2.50–2.40 (m, 1 H, CHH), 2.09–1.89 (m, 2 H, 2 CHH), 1.75–1.35
(m, 21 H, 21 CHH), 1.12 (s, 6 H, 2 CH3), 0.99 (s, 3 H, CH3) ppm.
13C NMR (75 MHz, CDCl3): δ = 177.8, 166.6, 113.0, 111.4, 95.7,
90.7, 78.1, 76.9, 75.9, 75.2 74.7, 73.2, 55.7, 54.7, 54.4, 37.3, 36.4,
36.3, 34.6, 30.6, 29.0, 24.9 (2 C), 23.8, 23.7, 23.6, 23.4, 16.6, 16.5,
9.6 ppm. HRMS (ESI): calcd. for C30H44NaO10 [M + Na]+
587.2827; found 587.2831.
involved the removal of Bn and MOM groups using
TMSBr (TMS = trimethylsilyl) before the catalytic hydro-
genolytic removal of Cbz. Bruzik conditions[29] (neat
TMSBr, 30 min; then MeOH, 1 h; then EtSH, 1 h) cleanly
furnished the desired fully inositol-head-deprotected deriva-
tive, but removal of the remaining Cbz upon hydrogenolysis
again proved to be troublesome.[33] Nevertheless, valuable
information was obtained from the 1H NMR spectra of the
material before hydrogenolysis: the desired compound was
accompanied (less than 7%) by the corresponding free
amine, i.e., our final target compound. This led us to per-
form the same protocol with TMSBr but with an increased
reaction time (1 h). But at this point, decomposition was
again observed in the 1H NMR spectra while the Cbz peaks
were still partially present.[34] Finally, using the Prestwich
protocol,[12] 30 was exposed to a mixture of TMSBr/
CH2Cl2 for 2 h and then simply stirred in methanol to un-
eventfully furnish the target 1-O-(ω-aminolauryl)-I(4,5)P2
(i.e., 7). According to this result, and in compliance with
our original synthetic strategy, unsaturated ω-amino fatty
acids could be also used as the acyl partners.
Conclusions
We have presented an efficient synthetic scheme for the
preparation of a model 1-O-(ω-aminoacyl)-IP, the lauryl de-
17: Rf = 0.22 (25% EtOAc in hexanes). [α]D = +12.6 (c = 1, CHCl3).
rivative 31 (12 steps from known camphanyl ester 14 and 1H NMR (300 MHz, CDCl3): δ = 5.15 (t, J = 5.4 Hz, 1 H, 3-H),
4.86, 4.79 (ABq, J = 6.4 Hz, 2 H, OCH2O), 4.61 (t, J = 5.4 Hz, 1
H, 2-H), 4.32 (dd, J = 8.2, 5.4 Hz, 1 H, 1-H), 4.10 (dd, J = 9.0,
5.5 Hz, 1 H, 4-H), 3.84 (dd, J = 10.3, 8.2 Hz, 1 H, 6-H), 3.46 (dd,
J = 10.3, 9.0 Hz, 1 H, 5-H), 3.43 (s, 3 H, OCH3), 2.52–2.42 (m, 1
H, CHH), 2.07–1.87 (m, 2 H, 2 CHH), 1.73–1.36 (m, 21 H, 21
CHH), 1.12 (s, 3 H, CH3), 1.06 (s, 3 H, CH3), 1.03 (s, 3 H, CH3)
ppm. 13C NMR (75 MHz, CDCl3): δ = 177.8, 166.7, 113.1, 111.4,
95.6, 90.9, 78.0, 77.2, 75.9, 75.0, 74.9, 73.1, 55.7, 54.8, 54.3, 37.1,
36.5, 36.4, 34.3, 30.6, 28.8, 24.9 (2 C), 23.8, 23.7, 23.6, 23.5, 16.79,
16.75, 9.7 ppm. HRMS (ESI): calcd. for C30H44NaO10 [M + Na]+
587.2827; found 587.2831.
44% overall yield). The same synthetic strategy is expected
to be easily applicable to the preparation of other 1- and 3-
O-(ω-aminoacyl)-IPs, including all possible phosphorylated
IPs, with variable phosphorylation a the 3-, 4- and 5- (or
1-, 5- and 6-, respectively) positions, and also including de-
rivatives bearing fatty chains of different lengths and satu-
ration patterns. In view of the central role played by IPs
and PIPs in cellular life, the preparation of compounds like
these may facilitate the isolation and fluorescent localiza-
tion of related proteins.
4
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Versatile Synthesis of 1-O-(ω-Aminolauryl)-I(4,5)P2
2,3:4,5-Di-O-cyclohexylidene-6-O-methoxymethyl-myo-inositol (18):
Lithium hydroxide monohydrate (420 mg, 10 mmol) was added to
a solution of campanyl ester 16 (565 mg, 1 mmol) in a 1:1 mixture
of THF/H2O (40 mL). The reaction mixture was stirred at room
temperature for 1 h. Then, it was extracted with EtOAc (2ϫ
50 mL). The combined organic extracts were washed with brine
(40 mL), dried (Na2SO4), and concentrated under reduced pressure.
The residue was purified by flash column chromatography
(10Ǟ35% EtOAc in hexanes) to give alcohol 18 (378 mg, 98%) as
a white foam. Rf: = 0.37 (35% EtOAc in hexanes). [α]D = –11.2 (c
5-H), 3.00 (br. s, 2 H, 2 OH), 1.80–1.65 (m, 4 H, 4 CHH), 1.55–
1.30 (m, 6 H, 6 CHH), 1.09 (s, 9 H, tBu) ppm. 13C NMR (75 MHz,
CDCl3): δ = 135.90, 135.86, 133.9, 133.4, 129.7 (2 C), 127.5, 127.4,
110.3, 98.5, 83.4, 77.7, 75.5 (2 C), 72.5, 71.5, 55.7, 38.1, 34.7, 26.9,
25.0, 23.9, 23.7, 19.3 ppm. HRMS (ESI): calcd. for C30H42NaO7Si
[M + Na]+ 565.2592; found 565.2600.
4,5-Di-O-benzoyl-1-O-tert-butyldiphenylsilyl-2,3-O-cyclohexylidene-
6-O-methoxymethyl-myo-inositol (21): Diol 20 (460 mg, 0.85 mmol)
was dissolved in dry CH2Cl2 (1 mL) and dry pyridine (5 mL) under
an Ar atmosphere, and the mixture was cooled to 0 °C. BzCl
(250 μL, 2.1 mmol) was added dropwise, and the mixture was left
to stir at room temperature for 24 h. Then EtOAc (70 mL) was
added and the mixture was washed with saturated aqueous sodium
hydrogen carbonate (2ϫ 30 mL) and brine (30 mL), and then dried
(Na2SO4) and concentrated under reduced pressure. The residue
was purified by flash column chromatography (10% EtOAc in hex-
anes) to give dibenzoate 21 (610 mg, 96%) as a white foam. Rf =
1
= 3.4, CHCl3). H NMR (300 MHz, CDCl3): δ = 4.86, 4.79 (ABq,
J = 6.6 Hz, 2 H, OCH2O), 4.45 (dd, J = 7.2, 3.9 Hz, 1 H, 2-H),
4.32 (t, J = 7.3 Hz, 1 H, 3-H), 4.06 (dd, J = 10.6, 8.0 Hz, 1 H, 4-
H), 4.02 (dd, J = 8.5, 3.7 Hz, 1 H, 6-H), 3.95 (t, J = 3.8 Hz, 1 H,
1-H), 3.45 (dd, J = 10.6, 8.4 Hz, 1 H, 5-H), 3.44 (s, 3 H, OCH3),
2.90 (br. s, 1 H, OH), 1.75–1.55 (m, 16 H, 16 CHH), 1.47–1.32 (m,
4 H, 4 CHH) ppm. 13C NMR (75 MHz, CDCl3): δ = 112.8, 111.0,
95.8, 77.9, 77.5, 77.2, 76.7, 75.5, 72.6, 55.6, 36.7, 36.42, 36.37, 33.9,
24.91, 24.88, 23.7, 23.6, 23.5, 23.3 ppm. HRMS (ESI): calcd. for
C20H32NaO7 [M + Na]+ 407.2040; found 407.2044.
1
0.54 (20% EtOAc in hexanes). [α]D = +16.4 (c = 4.5, CHCl3). H
NMR (300 MHz, CDCl3): δ = 7.93 (d, J = 7.3 Hz, 2 H, ArH), 7.89
(d, J = 7.3 Hz, 2 H, ArH), 7.84 (d, J = 6.8 Hz, 2 H, ArH), 7.74 (d,
J = 6.8 Hz, 2 H, ArH), 7.48–7.23 (m, 12 H, ArH), 6.00 (br. t, J =
8.7 Hz, 1 H, 4-H), 5.21 (dd, J = 9.6, 6.1 Hz, 1 H, 5-H), 4.73, 4.59
(ABq, J = 6.4 Hz, 2 H, OCH2O), 4.33 (dd, J = 7.1, 3.8 Hz, 1 H,
2-H), 4.19 (t, J = 7.1 Hz, 1 H, 3-H), 4.12 (dd, J = 7.1, 6.4 Hz, 1
H, 1-H), 3.98 (t, J = 6.5 Hz, 1 H, 6-H), 3.05 (s, 3 H, OCH3), 1.99–
1.70 (m, 4 H, 4 CHH), 1.51–1.30 (m, 6 H, 6 CHH), 1.16 (s, 9 H,
tBu) ppm. 13C NMR (75 MHz, CDCl3): δ = 165.8, 165.4, 136.2,
135.9, 133.5, 133.4, 132.9, 132.7, 130.0, 129.9, 129.8, 129.7, 129.6,
128.3, 128.14, 128.07, 127.7, 127.5, 111.0, 97.5, 77.7, 75.1, 74.7,
74.2 (2 C), 71.3, 55.8, 37.1, 34.3, 27.1, 25.1, 24.0, 23.8, 19.3 ppm.
HRMS (ESI): calcd. for C44H50NaO9Si [M + Na]+ 773.3116; found
773.3129.
1-O-tert-Butyldiphenylsilyl-2,3:4,5-di-O-cyclohexylidene-6-O-meth-
oxymethyl-myo-inositol (19): Alcohol 18 (430 mg, 1.1 mmol) and
imidazole (140 mg, 2.1 mmol) were dissolved in dry pyridine
(5 mL) under an Ar atmosphere. TBDPSCl (400 μL, 1.5 mmol) was
added dropwise, and the mixture was left to stir at 65 °C for 5 d.
Then, brine (30 mL) was added, and the mixture was stirred for
15 min and then extracted with EtOAc (3ϫ 50 mL). The combined
organic extracts were washed with brine (30 mL), dried (Na2SO4),
and concentrated under reduced pressure. The residue was purified
by flash column chromatography (10Ǟ30% EtOAc in hexanes) to
give silyl ether 19 (660 mg, 97%) as a thick pale yellow oil. Rf =
0.57 (25% EtOAc in hexanes). [α]D = +12.2 (c = 2, CHCl3). 1H
NMR (300 MHz, CDCl3): δ = 7.80–7.73 (m, 4 H, ArH), 7.42–7.32
(m, 6 H, ArH), 4.54–4.51 (ABq, J = 6.4 Hz, 2 H, OCH2O), 4.14–
4.01 (m, 3 H, 2-H, 3-H, 4-H), 3.94 (dd, J = 8.3, 6.0 Hz, 1 H, 6-H),
3.93 (br. s, 1 H, 2-H), 3.31 (s, 3 H, OCH3), 3.26 (t, J = 8.9 Hz, 1
H, 5-H), 1.73–1.25 (m, 20 H, 20 CHH), 1.12 (s, 9 H, tBu) ppm.
13C NMR (75 MHz, CDCl3): δ = 136.1, 136.0, 133.7, 133.6, 129.8,
129.7, 127.7, 127.4, 112.3, 110.8, 96.2, 77.9, 77.8, 77.3, 76.0, 75.7,
74.6, 55.6, 37.3, 36.6, 36.3, 34.1, 27.1, 25.1, 25.0, 23.8, 23.7, 23.68,
23.6, 16.3 ppm. HRMS (ESI): calcd. for C36H50NaO7Si [M +
Na]+ 645.3218; found 645.3208.
4,5-Di-O-benzoyl-1-O-tert-butyldiphenylsilyl-6-O-methoxymethyl-
myo-inositol (22): Acetal 21 (525 mg, 0.7 mmol) was dissolved in
CH2Cl2 (8 mL) and the solution was cooled to –40 °C. Aqueous
TFA (90%; 4 mL) was added dropwise and the mixture was left to
stir at –20 °C for 2 h. Then saturated aqueous sodium hydrogen
carbonate (20 mL) was added. The resulting slurry was stirred at
room temperature for 15 min and then extracted with CH2Cl2 (2ϫ
30 mL). The combined organic extracts were washed with brine
(20 mL), dried (Na2SO4), and concentrated under reduced pressure.
The residue was purified by flash column chromatography
(10Ǟ50% EtOAc in hexanes) to give diol 22 (440 mg, 94%) as a
white foam. Rf = 0.20 (25% EtOAc in hexanes). [α]D = +15.7 (c =
2.8, CHCl3). 1H NMR (300 MHz, CDCl3): δ = 7.97 (br. d, J =
7.1 Hz, 2 H, ArH), 7.85 (br. d, J = 7.1 Hz, 2 H, ArH), 7.76 (br. d,
J = 6.3 Hz, 4 H, ArH), 7.40–7.23 (m, 12 H, ArH), 5.67 (t, J =
9.9 Hz, 1 H, 4-H), 5.40 (t, J = 9.8 Hz, 1 H, 5-H), 4.88 (d, J =
6.8 Hz, 1 H, OCHHO), 4.51 (d, J = 6.8 Hz, 1 H, OCHHO), 4.31
(t, J = 9.5 Hz, 1 H, 6-H), 3.97 (dd, J = 9.2, 2.5 Hz, 1 H, 3-H), 3.72
(br. s, 1 H, 2-H), 3.49 (br. d, J = 9.6 Hz, 1 H, 1-H), 3.13 (br. s, 1
H, OH), 2.97 (br. s, 1 H, OH), 2.97 (s, 3 H, OCH3), 1.12 (s, 9 H,
tBu) ppm. 13C NMR (75 MHz, CDCl3): δ = 166.9, 165.9, 135.8,
135.6, 133.5, 133.0, 132.8, 132.6, 130.1, 130.0, 129.9, 129.7, 129.6,
129.3, 128.1 (2 C), 127.9, 127.8, 98.8, 78.7, 74.3, 74.2, 72.4, 72.0,
70.6, 55.8, 27.0, 19.3 ppm. HRMS (ESI): calcd. for C38H43O9Si [M
+ H]+ 671.2671; found 671.2666.
1-O-tert-Butyldiphenylsilyl-2,3-O-cyclohexylidene-6-O-methoxy-
methyl-myo-inositol (20): Acetal 19 (455 mg, 0.75 mmol) was dis-
solved in a 5:1 mixture of CH2Cl2/MeOH (48 mL), and the mixture
was cooled to 0 °C. AcCl (80 μL, 1.15 mmol) was added dropwise,
and the mixture was left to stir at 0 °C for 90 min. Then saturated
aqueous sodium hydrogen carbonate (6 mL) was added, and the
mixture was vigorously stirred at 0 °C for 15 min and then ex-
tracted with CH2Cl2 (5ϫ 30 mL). The combined organic extracts
were washed with brine (20 mL), dried (Na2SO4), and concentrated
under reduced pressure. The residue was purified by flash column
chromatography (10 Ǟ 50 % EtOAc in hexanes) to give diol 20
(402 mg, 99%) as a sticky white foam. Rf = 0.19 (50% EtOAc in
hexanes). [α]D = +26.4 (c = 3.8, CHCl3). 1H NMR (300 MHz,
CDCl3): δ = 7.79–7.72 (m, 4 H, ArH), 7.41–7.32 (m, 6 H, ArH),
4.62, 4.09 (ABq, J = 6.6 Hz, 2 H, OCH2O), 3.94 (dd, J = 8.7,
4.1 Hz, 1 H, 3-H), 3.88 (t, J = 4.3 Hz, 1 H, 2-H), 3.72–3.65 (m, 3
H, 1-H, 4-H, 6-H), 3.38 (s, 3 H, OCH3), 3.12 (t, J = 9.2 Hz, 1 H, 4,5-Di-O-benzoyl-1-O-tert-butyldiphenylsilyl-2,3,6-tri-O-methoxy-
methyl-myo-inositol (23): Diol 22 (335 mg, 0.5 mmol) was dissolved
in dry DMF (10 mL) and DIPEA (3.6 mL) under an Ar atmo-
sphere. MOMCl (1.4 mL, 18 mmol) was added dropwise at room
temperature, and the mixture was stirred at 75 °C for 4 d. Then the
Eur. J. Org. Chem. 0000, 0–0
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Pages: 9
A. E. Koumbis et al.
FULL PAPER
reaction mixture was cooled to room temperature and saturated
aqueous sodium hydrogen carbonate (20 mL) was added. The re-
sulting slurry was stirred at room temperature for 15 min and then
extracted with CH2Cl2 (3ϫ 50 mL). The combined organic extracts
were washed with brine (30 mL), dried (Na2SO4), and concentrated
under reduced pressure. The residue was purified by flash column
chromatography (10Ǟ20% EtOAc in hexanes) to give MOM ether
23 (378 mg, 100%) as a pale yellow foam. Rf = 0.46 (25% EtOAc
dry THF (4 mL) under an Ar atmosphere. TBAF solution in THF
(1 m; 180 μL, 0.36 mmol) was added dropwise, and the mixture was
stirred at room temperature for 1 h. Then it was quenched by the
addition of saturated aqueous ammonium chloride (2 mL). EtOAc
(10 mL) was added, and the resulting slurry was vigorously stirred
for 15 min, and then extracted with CH2Cl2 (3ϫ 10 mL). The com-
bined organic extracts were washed with brine (10 mL), dried
(Na2SO4), and concentrated under reduced pressure. The residue
in hexanes). [α]D = +33.1 (c = 2.9, CHCl3). 1H NMR spectrum was was purified by flash column chromatography (10Ǟ35% EtOAc
identical with that reported in the literature.[29] 13C NMR (75 MHz,
CDCl3): δ = 165.8, 165.4, 135.9, 135.8, 133.8, 132.8, 132.7, 130.04,
in hexanes) to give alcohol 26 (231 mg, 93%) as a pale yellow
amorphous solid. Rf = 0.20 (35% EtOAc in hexanes). [α]D = –41.8
130.97, 129.9, 129.7, 129.6, 129.5, 128.09 (2 C), 128.06, 127.9, (c = 2.0, CHCl3). 1H and 31P NMR spectra were identical with
127.7, 98.5, 97.4, 94.4, 78.3, 75.1, 74.2, 73.3, 72.8, 72.3, 55.8, 55.6, those reported in the literature.[29] 13C NMR (75 MHz, CDCl3): δ
3
3
55.3, 27.1, 19.2 ppm. HRMS (ESI): calcd. for C42H50NaO11Si [M = 135.91 (d, JCP = 8.0 Hz, 2 C), 135.87 (d, JCP = 8.0 Hz), 135.8
+ Na]+ 781.3015; found 781.3019.
(d, JCP = 7.5 Hz), 128.4, 128.3, 128.2, 128.14, 128.10, 128.06,
3
127.9, 127.8 127.73, 127.69, 98.9, 97.5, 96.7, 83.2, 79.2 (t, JCP
=
=
1-O-tert-Butyldiphenylsilyl-2,3,6-tri-O-methoxymethyl-myo-inositol
(24): Following a slightly modified procedure,[29] dibenzoate 23 gave
2
5.5 Hz), 77.8 (t, JCP = 5.8 Hz), 75.4, 74.9, 70.3, 69.4 (d, JCP
2
2
6.3 Hz), 69.3 (d, JCP = 6.9 Hz), 69.0 (d, JCP = 5.2 Hz, 2 C), 55.8,
55.6, 55.5 ppm. HRMS (ESI): calcd. for C40H50NaO15P2 [M +
Na]+ 855.2517; found 855.2525.
diol 24 in 92% yield. Rf = 0.13 (50% EtOAc in hexanes). [α]D
=
+80.6 (c = 2, CHCl3). 1H and 13C NMR spectra were identical
with those reported in the literature.[29] HRMS (ESI): calcd. for
C28H43O9Si [M + H]+ 551.2671; found 551.2661.
Methyl 12-(Benzyloxycarbonylamino)dodecanoate (28): Amine 27
(500 mg, 2.2 mmol)[31] was dissolved in dry CH2Cl2 (17 mL) under
an Ar atmosphere. DMAP (53 mg, 0.43 mmol) and dry Et3N
(1.5 mL, 11 mmol) were added, and the resulting solution was co-
oled to 0 °C. Then, CbzCl (0.37 mL, 2.6 mmol) was added drop-
wise, and the mixture was left to stir at room temperature for 12 h.
H2O (10 mL) was added, and the resulting mixture was vigorously
stirred for 15 min. The organic phase was washed with brine (2ϫ
10 mL), and the combined aqueous washes were re-extracted with
CH2Cl2 (15 mL). Finally, the combined organic extracts were dried
(Na2SO4) and concentrated under reduced pressure. The residue
was purified by flash column chromatography (10Ǟ50% EtOAc
in hexanes) to give protected amine 28 (705 mg, 89%) as a thick
colorless oil. Rf = 0.60 (50% EtOAc in hexanes). 1H NMR
(300 MHz, CDCl3): δ = 7.35–7.29 (m, 5 H, ArH), 5.09 (s, 2 H,
CH2Ph), 4.78 (br. s, 1 H, NH), 3.65 (s, 3 H, OCH3), 3.17 (q, J =
6.6 Hz, 2 H, NHCH2), 2.29 (t, J = 7.5 Hz, 2 H, COCH2), 1.64–
1.56 (m, 2 H, COCH2CH2), 1.50–1.46 (m, 2 H, NHCH2CH2), 1.27
(br. s, 14 H, 7 CH2CH2CH2) ppm. 13C NMR (75 MHz, CDCl3): δ
= 174.2, 156.4, 136.7, 128.4, 128.0, 66.5, 51.3, 41.1, 34.0, 29.9,
29.40, 29.39, 29.3, 29.2, 29.14, 29.07, 26.7, 24.9 ppm. HRMS (ESI):
calcd. for C21H33NNaO4 [M + Na]+ 386.2302; found 386.2310.
Tetrabenzyl 4,5-(1-O-tert-Butyldiphenylsilyl-2,3,6-tri-O-methoxy-
methyl-myo-inosityl) Bisphosphate (25): Diol 24 was phosphory-
lated following a modification of a known procedure.[29] Thus,
starting diol 24 (260 mg, 0.47 mmol) was dried under high vacuum
for 24 h. Then a solution of 1H-tetrazole in acetonitrile (0.45 m;
10 mL, 4.5 mmol) was added under a N2 atmosphere at room tem-
perature. Dropwise addition (30 min) of dibenzyl N,N-diisoprop-
ylphosphoramidite (0.5 mL, 1.5 mmol) to the resulting suspension
followed. The resulting slurry was vigorously stirred at room tem-
perature for 24 h. CH2Cl2 (5 mL) was added and the mixture was
cooled to –40 °C. A solution of m-chloroperbenzoic acid (70%;
740 mg, 3 mmol) in CH2Cl2 (5 mL) was added dropwise, and the
mixture was left to stir at 0 °C for 3 h. Then the mixture was diluted
with CH2Cl2 (50 mL) and successively washed with aqueous so-
dium sulfite (10%; 2ϫ 30 mL), saturated aqueous sodium hydro-
gen carbonate (2ϫ 25 mL), H2O (25 mL), and brine (25 mL). The
organic phase was dried (Na2SO4), and the solvents were removed
under reduced pressure (30 °C). The resulting residue was purified
by flash column chromatography (10Ǟ50% EtOAc in hexanes) to
give bisphosphate 25 (445 mg, 88%) as a colorless thick oil. Rf =
0.45 (50% EtOAc in hexanes). [α]D = +7.0 (c = 2.6, CHCl3). 1H
NMR (300 MHz, CDCl3): δ = 7.78 (br. d, J = 6.5 Hz, 2 H, ArH), 12-(Benzyloxycarbonylamino)dodecanoic Acid (29): Ester 28
7.71 (d, J = 6.0 Hz, 2 H, ArH), 7.43–7.24 (m, 26 H, ArH), 5.14– (580 mg, 1.6 mmol) was dissolved in a 3:1 mixture of THF/H2O
4.93 (m, 9 H, 4 OCH2Ph, OCHHO), 4.79 (q, 3JHH = 3JHP = 9.9 Hz, (12 mL). LiOH monohydrate (202 mg, 4.8 mmol) was added, and
1 H, 4-H), 4.76 (d, J = 6.1 Hz, 1 H, OCHHO), 4.59, 4.52 (ABq, J the resulting slurry was left to stir vigorously at room temperature
3JHP = 9.4 Hz, 1 H, 5- for 10 h. Then the mixture was acidified to pH 2 using HCl (1 m).
H), 4.28, 4.05 (ABq, J = 7.1 Hz, 2 H, OCH2O), 4.19 (t, J = 9.6 Hz, CH2Cl2 (25 mL) was added, and the organic phase was washed
3
= 6.4 Hz, 2 H, OCH2O), 4.32 (q, JHH
=
1 H, 6-H), 3.87 (br. d, J = 9.7 Hz, 1 H, 1-H), 3.38 (s, 3 H, OCH3), with brine (2ϫ 10 mL). The combined aqueous washes were re-
3.29 (br. s, 1 H, 2-H), 3.26 (s, 3 H, OCH3), 3.20 (br. d, J = 9.8 Hz, extracted with CH2Cl2 (10 mL). Finally, the combined organic ex-
1 H, 3-H), 2.97 (s, 3 H, OCH3), 1.11 (s, 9 H, tBu) ppm. 13C NMR
(75 MHz, CDCl3): δ = 136.21 (d, JCP = 7.5 Hz, 3 C), 136.17 (d, sure. The residue was purified by flash column chromatography
3JCP = 8.0 Hz), 136.0, 135.8, 133.9, 132.6, 130.0, 129.8, 128.30, (20Ǟ60% EtOAc in hexanes) to give acid 29 (530 mg, 95%) as a
128.25, 128.2, 128.10, 128.05, 128.03, 128.00, 127.91, 127.88, 127.8, white waxy solid. Rf = 0.16 (25% EtOAc in hexanes). 1H NMR
tracts were dried (Na2SO4) and concentrated under reduced pres-
3
127.72, 127.67, 98.7, 97.3, 95.9, 78.5 (t, JCP = 5.2 Hz), 77.8 (q, JCP
spectrum was identical with that reported in the literature.[30] 13
C
NMR (75 MHz, CDCl3): δ = 178.1, 156.8, 137.0, 128.5, 128.0, 66.7,
= 2.9 Hz), 75.8 (br. s), 75.5, 74.2, 73.7, 69.5 (d, 2JCP = 5.7 Hz), 69.3
(d, 2JCP = 5.7 Hz), 69.2 (d, 2JCP = 5.2 Hz), 69.0 (d, 2JCP = 5.2 Hz), 41.4, 33.9, 30.0, 29.4, 29.32, 29.26, 29.2, 29.1, 29.0, 26.7, 14.7 ppm.
56.9, 55.6, 55.4, 27.2, 19.1 ppm. 31P NMR (100 MHz, CDCl3, 31P– HRMS (ESI): calcd. for C20H31NNaO4 [M + Na]+ 372.2145; found
1H decoupled): δ = –1.23, –1.51 ppm. HRMS (ESI): calcd. for
372.2148.
C56H68NaO15P2Si [M + Na]+ 1093.3695; found 1093.3679.
Tetrabenzyl 4,5-{1-O-[12-N-(Benzyloxycarbonyl)aminododecanoyl]-
Tetrabenzyl 4,5-(2,3,6-Tri-O-methoxymethyl-myo-inosityl) Bis-
phosphate (26): Silyl ether 25 (320 mg, 0.3 mmol) was dissolved in
2,3,6-tri-O-methoxymethyl-myo-inosityl}
Alcohol 25 (50 mg, 0.06 mmol) and carboxylic acid 29 (63 mg,
Bisphosphate
(30):
6
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Eur. J. Org. Chem. 0000, 0–0

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Versatile Synthesis of 1-O-(ω-Aminolauryl)-I(4,5)P2
0.18 mmol) were dissolved in dry CH2Cl2 (5 mL) under an Ar at-
mosphere. A catalytic amount of DMAP (1 mg) was added, fol-
lowed by DCC (37 mg, 0.18 mmol). The mixture was stirred at
room temperature for 3 d. Then it was concentrated under reduced
pressure, and the residue was triturated with a mixture of CH2Cl2/
Et2O (9:1). The crystallized urea was removed by filtration and the
filtrate was concentrated to give a residue, which was further puri-
fied by flash column chromatography (40Ǟ70% EtOAc in hex-
anes) to give ester 30 (67 mg, 96%) as a colorless thick oil. Rf =
Acknowledgments
This work was partially supported by the 03ED402 research pro-
ject, implemented within the framework of the Reinforcement Pro-
gram of Human Research Manpower (PENED) and co-financed
by National and Community Funds 25% from the (Greek Ministry
of Development, General Secretariat of Research and Technology
and 75% from EU European Social Fund).
1
0.27 (65% EtOAc in hexanes). [α]D = –27.5 (c = 0.36, CHCl3). H
NMR (300 MHz, CDCl3): δ = 7.36–7.25 (m, 25 H, ArH), 5.15–
[1] a) D. C. Billington, The Inositol Phosphates, Wiley-VCH,
Weinheim, Germany, 1993; b) B. V. L. Potter, D. Lampe, An-
gew. Chem. 1995, 107, 2085; Angew. Chem. Int. Ed. Engl. 1995,
34, 1933–1972; c) M. A. J. Ferguson, S. W. Homans, R. A.
Dwek, T. W. Rademacher, Science 1988, 239, 753–759.
[2] G. D. Prestwich, Chem. Biol. 2004, 11, 619–637.
[3] Selected publications: a) M. J. Berridge, Nature 1993, 365, 388–
389; b) A. Toker, L. C. Cantley, Nature 1997, 387, 673–676; c)
S. Corvera, M. P. Czech, Trends Cell Biol. 1998, 8, 442–446; d)
L. V. Dekker, A. W. Segal, Science 2000, 287, 982–985; e) M.
Overduin, M. L. Cheever, T. G. Kutateladze, Mol. Interventions
2001, 1, 150–159; f) A. Toker, Cell Mol. Life Sci. 2002, 59, 761–
779; g) C. Pendaries, H. Tronchére, M. Plantavid, B. Payrastre,
FEBS Lett. 2003, 546, 25–31.
3
4.94 (m, 10 H, 5 OCH2Ph), 4.90 (q, JHH = 3JCP = 9.4 Hz, 1 H, 4-
H), 4.81 (dd, J = 10.3, 2.3 Hz, 1 H, 1-H), 4.77, 4.64 (ABq, J =
6.7 Hz, 2 H, OCH2O), 4.68 (br. s, 1 H, NH), 4.62, 4.57 (ABq, J =
6.9 Hz, 2 H, OCH2O), 4.57, 4.47 (ABq, J = 6.9 Hz, 2 H, OCH2O),
3
4.42 (q, JHH
=
3JCP = 9.3 Hz, 1 H, 5-H), 4.13 (br. s, 1 H, 2-H),
4.05 (t, J = 9.7 Hz, 1 H, 6-H), 3.61 (dd, J = 10.1, 1.8 Hz, 1 H, 3-
H), 3.38 (s, 3 H, OCH3), 3.26 (s, 3 H, OCH3), 3.22 (s, 3 H, OCH3),
3.17 (q, J = 6.6 Hz, 2 H, NHCH2), 2.35 (t, J = 6.4 Hz, 2 H,
COCH2), 1.68–1.58 (m, 2 H, COCH2CH2), 1.51–1.46 (m, 2 H,
NHCH2CH2), 1.26 (br. s, 14 H, 7 CH2CH2CH2) ppm. 13C NMR
(75 MHz, CDCl3): δ = 172.8, 156.4, 136.8, 136.3–136.0 (m, 4 C),
128.52, 128.50, 128.44, 128.41, 128.38, 128.33, 128.29, 128.2, 128.1,
128.0, 127.9, 99.0, 97.5, 97.2, 79.3 (t, JCP = 5.5 Hz), 77.5–77.2 (m,
[4] a) M. J. Berridge, R. F. Irvine, Nature 1989, 341, 197–205; b)
M. J. Berridge, Nature 1993, 361, 315–325.
3
3
2 C), 75.3, 74.3, 71.5, 69.8 (d, JCP = 5.9 Hz), 69.6 (d, JCP
=
5.9 Hz), 69.5 (d, 3JCP = 5.1 Hz), 69.3 (d, 3JCP = 5.1 Hz), 66.6, 56.2,
55.9, 55.7, 41.2, 34.2, 30.0, 29.7, 29.5, 29.4, 29.3, 29.2 (2 C), 26.7,
24.5 ppm. 31P NMR (100 MHz, CDCl3, 31P–1H decoupled): δ =
–1.34, –1.46 ppm. HRMS (ESI): calcd. for C60H79NNaO18P2 [M +
Na]+ 1186.4665; found 1186.4658.
[5] A. M. Shamsuddin, I. Vucenik, K. E. Cole, Life Sci. 1997, 61,
343–354.
[6] a) P. M. Hwang, A. Verma, D. S. Bredt, S. H. Snyder, Proc.
Natl. Acad. Sci. USA 1990, 87, 7395–7399; b) I. Boekhoff, E.
Tareilus, J. Strotmann, H. Breer, , EMBO J. 1990, 9, 2543–
2458; c) D. A. Fadool, B. W. Ache, Neuron 1992, 9, 1–20; d)
G. V. Ronnett, S. H. Snyder, Trends Neurosci. 1992, 15, 508–
513; e) R. C. Bruch, Comp. Biochem. Physiol., Part B 1996, 113,
451–459.
[7] a) P. A. Janmey, Annu. Rev. Physiol. 1994, 56, 169–191; b) A.
Lambrechts, J. Van Damme, M. Goethals, J. Vandekerckhove,
C. Ampe, Eur. J. Biochem. 1995, 230, 281–286; c) P. A. Janmey,
Chem. Biol. 1995, 2, 61–65.
[8] a) L. Stephens, T. Jackson, P. T. Hawkins, J. Biol. Chem. 1993,
268, 17162–17172; b) L. R. Stephens, T. R. Jackson, P. T. Haw-
kins, Biochim. Biophys. Acta Mol. Cell Res. 1993, 1179, 27–75;
c) L. Stephens, F. T. Cooke, R. Walters, T. Jackson, S. Volinia,
I. Gout, M. D. Waterfield, P. T. Hawkins, Curr. Biol. 1994, 4,
203–214.
[9] S. J. Conway, G. J. Miller, Nat. Prod. Rep. 2007, 24, 687–707.
[10] Y. J. Xu, B. R. Sculimbrene, S. J. Miller, J. Org. Chem. 2006,
71, 4919–4928.
Disodium 4,5-[1-O-(12-Aminododecanoyl)-myo-inosityl] Bisphos-
phate (31): TMSBr (ca. 0.4 mL) was distilled into a flask cooled to
–78 °C containing a solution of ester 30 (23 mg, 0.02 mmol) in dry
CH2Cl2 (0.6 mL) under an Ar atmosphere. The mixture was left to
stir in the dark for 2 h. Then it was concentrated under reduced
pressure at room temperature, and the residue was placed under
high vacuum for 6 h. MeOH (1 mL) was added, and the mixture
was stirred for 1 h at room temperature. Evaporation under re-
duced pressure at room temperature and washing with CHCl3 (3ϫ
1 mL) yielded a waxy solid. Aqueous sodium hydrogen carbonate
(0.1 m; 0.4 mL, 0.04 mmol) was added along with tBuOH (1 mL),
and the mixture was concentrated under reduced pressure at room
temperature. Additional quantities of tBuOH (2ϫ 1 mL) were used
to azeotropically remove traces of water by evaporation under re-
duced pressure at room temperature. The residue was further dried
under high vacuum for 12 h to give sodium phosphate 31 (9.4 mg,
81%) as an amorphous white solid. [α]D = +2.1 (c = 0.15, H2O).
[11] T. Shirai, K. Tanaka, Y. Terada, T. Sawada, R. Shirai, Y. Hash-
imoto, S. Nagata, A. Iwamatsu, K. Okawa, S. Li, S. Hattori,
H. Mano, Y. Fukui, Biochim. Biophys. Acta Mol. Cell Res.
1998, 1402, 292–302.
1H NMR (acid form, 300 MHz, CD3OD): δ = 4.75 (obscured, 1
[12] H. L. Zhang, N. Markadieu, R. Beauwens, C. Erneux, G. D.
Prestwich, J. Am. Chem. Soc. 2006, 128, 16464–16465.
[13] Y. Xu, S. A. Lee, T. G. Kutateladze, D. Sbrissa, A. Shisheva,
G. D. Prestwich, J. Am. Chem. Soc. 2006, 128, 885–897.
[14] a) R. J. Kubiak, K. S. Bruzik, Bioorg. Med. Chem. Lett. 1997,
7, 1231–1234; b) C. Mihai, J. Mataka, S. Riddle, M. D. Tsai,
K. S. Bruzik, Bioorg. Med. Chem. Lett. 1997, 7, 1235–1238; c)
H. Zhang, Y. Xu, N. Markadieu, R. Beauwens, C. Erneux,
G. D. Prestwich, Bioorg. Med. Chem. Lett. 2008, 18, 762–766.
[15] a) J. Chen, G. D. Prestwich, J. Org. Chem. 1998, 63, 430–431;
b) H. L. Zhang, Y. Xu, Z. Zhang, E. R. Liman, G. D. Prest-
wich, J. Am. Chem. Soc. 2006, 128, 5642–5643.
[16] a) K. K. Reddy, M. Saady, J. R. Falck, G. Whited, J. Org.
Chem. 1995, 60, 3385–3390; b) A. Nishikawa, S. Saito, Y. Hash-
imoto, K. Koga, R. Shirai, Tetrahedron Lett. 2001, 42, 9195–
9198; c) K. J. Kayser-Bricker, P. A. Jordan, S. J. Miller, Tetrahe-
dron 2008, 64, 7015–7020.
3
H, 1-H), 4.50 (q, JHH
=
3JCP = 8.2 Hz, 1 H, 4-H), 4.18 (br. q,
3JHH = JCP = 9.2 Hz, 1 H, 5-H), 4.06 (br. s, 1 H, 2-H), 4.03 (t, J
= 9.6 Hz, 1 H, 6-H), 3.67 (br. d, J = 8.5 Hz, 1 H, 3-H), 2.91 (br.
q, J = 6.4 Hz, 2 H, NHCH2), 2.40 (t, J = 7.5 Hz, 2 H, COCH2),
1.64 (br. s, 4 H, COCH2CH2, NHCH2CH2), 1.33, 1.29 (2 br. s, 14
H, 7 CH2CH2CH2) ppm. 13C NMR (acid form, 75 MHz, CD3OD):
δ = 173.5, 79.9 (t, JCP = 5.1 Hz), 79.1 (br. s), 78.7, 77.4, 75.6, 72.1,
40.8, 35.1, 31.1, 30.12, 30.14, 30.0, 29.9, 29.8, 29.7, 26.6, 24.7 ppm.
31P NMR (acid form, 100 MHz, CD3OD, 31P–1H decoupled): δ =
3.39, 2.89 ppm. HRMS (ESI): calcd. for C18H35NNaO13P2 [M –
Na]– 558.1487; found 558.1491.
3
Supporting Information (see footnote on the first page of this arti-
cle): Copies of the 1H NMR and 13C NMR spectra for all com-
pounds.
Eur. J. Org. Chem. 0000, 0–0
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A. E. Koumbis et al.
FULL PAPER
[17] P. W. Rzepecki, G. D. Prestwich, J. Org. Chem. 2002, 67, 5454–
5460.
[18] a) J. Chen, A. A. Profit, G. D. Prestwich, J. Org. Chem. 1996,
61, 6305–6312; b) O. Thum, J. Chen, G. D. Prestwich, Tetrahe-
dron Lett. 1996, 37, 9017–9020.
[19] G. D. Prestwich, Acc. Chem. Res. 1996, 29, 503–513, and refer-
ences cited therein.
J.-M. Lehn, C. Nicolau, ChemBioChem 2011, 12, 777–783;
d) K. C. Fylaktakidou, C. D. Duarte, A. E. Koumbis, C.
Nicolau, J.-M. Lehn, ChemMedChem 2011, 6, 153–168;
e) A. E. Koumbis, C. D. Duarte, C. Nicolau, J.-M. Lehn,
ChemMedChem 2011, 6, 169–180; f) K. C. Fylaktakidou, C. D.
Duarte, R. Jogireddy, A. E. Koumbis, C. Nicolau, J.-M. Lehn,
ChemMedChem 2011, 6, 1495–1508.
[20] R. J. Anderson, S. L. Osborne, F. A. Meunier, G. F. Painter, J.
Org. Chem. 2010, 75, 3541–3551.
[26]
[27]
[28]
S. C. Ghosh, B. Kim, J. Im, W. S. Lee, C.-N. Im, Y.-T. Chang,
W. Kim, K.-T. Kim, S.-K. Chung, Bull. Korean Chem. Soc.
2010, 31, 3623–3631.
J. P. Vacca, S. J. deSolms, J. R. Huff, D. C. Billington, R. Baker,
J. J. Kulagowski, I. M. Mawer, Tetrahedron 1989, 45, 5679–
5702.
S. Iacobucci, M. d’Ararcao, Synth. Commun. 1994, 24, 809–
817.
R. J. Kubiak, K. S. Bruzik, J. Org. Chem. 2003, 68, 960–968.
N. Kameta, G. Mizuno, M. Masuda, H. Minamikawa, M. Ko-
[21] a) G. F. Painter, J. W. Thuring, Z.-Y. Lim, A. B. Holmes, P. T.
Hawkins, L. R. Stephens, Chem. Commun. 2001, 645–646; b)
Z.-Y. Lim, J. W. Thuring, A. B. Holmes, M. Manifava, N. T.
Ktistakis, J. Chem. Soc. Perkin Trans. 1 2002, 1067–1075; c)
S. J. Conway, J. Gardiner, S. J. A. Grove, M. K. Johns, Z.-Y.
Lim, G. F. Painter, D. E. J. E. Robinson, C. Schieber, J. W.
Thuring, L. S.-M. Wong, M.-X. Yin, A. W. Burgess, B. Cati-
mel, P. T. Hawkins, N. T. Ktistakis, L. R. Stephens, A. B.
Holmes, Org. Biomol. Chem. 2010, 8, 66–76.
[22] T. Sawada, R. Shirai, Y. Matsuo, Y. Kabuyama, K. Kimura, Y.
Fukui, Y. Hashimoto, S. Iwasaki, Bioorg. Med. Chem. Lett.
1995, 5, 2263–2266.
[23] K. Tanaka, S. Imajoh-Ohmi, T. Sawada, R. Shirai, Y. Hashim-
oto, S. Iwasaki, K. Kaibuchi, Y. Kanaho, T. Shirai, Y. Terada,
K. Kimura, S. Nagata, Y. Fukui, Eur. J. Biochem. 1997, 245,
512–519.
[24] D. J. R. Massy, P. Wyss, Helv. Chim. Acta 1990, 73, 1037–1057.
[25] a) K. C. Fylaktakidou, J. M. Lehn, R. Greferath, C. Nicolau,
Bioorg. Med. Chem. Lett. 2005, 15, 1605–1608; b) A. Biolo, R.
Greferath, D. A. Siwik, F. Qin, E. Valsky, K. C. Fylaktakidou,
S. Pothukanuri, C. D. Duarte, R. P. Schwartz, J.-M. Lehn, C.
Nicolau, W. S. Colucci, Proc. Natl. Acad. Sci. USA 2009, 106,
1926–1929; c) M. Aprahamian, G. Bour, C. Y. Akladios, K. C.
Fylaktakidou, R. Greferath, L. Soler, J. Marescaux, J.-M. Egly,
[29]
[30]
giso, T. Shimizu, Chem. Lett. 2007, 36, 896–897.
[31] C. M. Jakobsen, S. R. Denmeade, J. T. Isaacs, A. Gady, C. E.
Olsen, S. B. Christensen, J. Med. Chem. 2001, 44, 4696–4703.
[32] In situ formation of the corresponding sodium or triethylam-
monium salts would not allow the subsequent removal of
MOM groups.
[33] The reason for this failure remains unclear. All different solvent
systems used gave partial decomposition, even in the presence
of 2 equiv. NaHCO3.
[34] As expected, removal of the Cbz group not only resulted in
disappearance of the corresponding peaks in the 1H NMR
spectra, but also caused significant shielding of certain methyl-
ene protons (on C-12 of the lauric moiety).
Received: May 30, 2012
Published Online: ᭿
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Eur. J. Org. Chem. 0000, 0–0

Job/Unit: O20726
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Date: 04-09-12 15:30:15
Pages: 9
Versatile Synthesis of 1-O-(ω-Aminolauryl)-I(4,5)P2
myo-Inositol Phosphates
A.-A. C. Varvogli, K. C. Fylaktakidou,
T. Farmaki, J. G. Stefanakis,
A. E. Koumbis* ................................. 1–9
Versatile Synthesis of 1-O-(ω-Amino-
lauryl)-I(4,5)P2
1-O-(ω-Aminolauryl)-I(4,5)P2 was pre-
pared from myo-inositol using a versatile
and high-yielding scheme. The same syn-
thetic strategy is expected to be easily appli-
cable to the preparation of other 1- and 3-
O-(ω-aminoacyl)-IPs, with potential use in
the isolation and localization of proteins.
Keywords: Bioorganic chemistry / Natural
products / Inositols / Phosphates / Fatty
acids / Lipids
Eur. J. Org. Chem. 0000, 0–0
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