Short and efficient synthesis of a stock material of dihydroxyacetone phosphate from glycidol

Article abstract of DOI:10.1016/j.tetlet.2004.08.148

As enzymatic syntheses are expensive for a large-scale preparation of DHAP, a precursor leading to DHAP was synthesized in three steps starting from (±) glycidol; the stable benzylated stock material afforded by hydrogenolysis DHAP in high purity, which may be used directly without purification in enzymatic aldol synthesis.

Full text of DOI:10.1016/j.tetlet.2004.08.148

Tetrahedron
Letters
Tetrahedron Letters 45 (2004) 7921–7923
Short and efficient synthesis of a stock material of
dihydroxyacetone phosphate from glycidol
Odile Meyer, Michel Rohmer and Catherine Grosdemange-Billiard*
´
Universite Louis Pasteur, CNRS UMR 7123, Institut Le Bel, 4 rue Blaise Pascal, 67070 Strasbourg Cedex, France
Received 8 June 2004;revised 18 August 2004;accepted 24 August 2004
Available online 11 September 2004
Abstract—As enzymatic syntheses are expensive for a large-scale preparation of DHAP, a precursor leading to DHAP was synthe-
sized in three steps starting from ( ) glycidol;the stable benzylated stock material afforded by hydrogenolysis DHAP in high purity,
which may be used directly without purification in enzymatic aldol synthesis.
Ó 2004 Elsevier Ltd. All rights reserved.
1. Introduction
molecules of triose phosphate, the glyceraldehyde 3-
phosphate and the DHAP. Two other enzymatic
methods have been developed. One proceeds via phos-
phorylation of free dihydroxyacetone using glycerol
kinase and ATP,5 and the other is the oxidation of the
L-glycerol 1-phosphate by oxygen in the presence of
the glycerophosphate oxidase coupled with a catalase
allowing the decomposition of hydrogen peroxide.6
Enzymatic syntheses are however too expensive to be
used for large-scale preparation of DHAP, and chemical
methods are preferentially used.
The stereoselective formation of C–C bonds is one of the
main goals in organic synthesis. Among many methods,
the aldolization reaction with a chiral auxilliary allows
to form diastereoselective and even enantioselective C–
C bonds.1 Aldolases are known as useful biocatalysts
to form these bonds and the enzymatic reaction takes
place with high enantio- and diastereoselectivity.1 Of
more than 20 known aldolases, D-fructose 1,6-biphos-
phate aldolase (EC 4.1.2.13), D-tagatose 1,6-biphos-
phate aldolase (EC 4.1.2.-), L-fucose 1-phosphate
aldolase (EC 4.1.2.17) and L-rhamnulose 1-phosphate
aldolase (EC 4.1.2.19) have demonstrated their synthetic
utility: these enzymes catalyze the asymmetric addition
of dihydroxyacetone phosphate (DHAP) as nucleophile
(donor) on a variety of aldehydes as electrophilic sub-
strate.2 Each enzyme leads to only one of the four pos-
sible stereoisomers (Scheme 1).3 Commercially available
DHAP remains too expensive to be used in organic syn-
thesis and therefore improvement of its preparation is
still a subject of interest.
The first chemical precursor of DHAP was the dihydr-
oxyacetone phosphate dimethyl ketal 1a (Scheme 2) syn-
thesized by Ballou and Fischer.7 Their synthesis led to
DHAP in eight steps with an overall yield of 13%. Other
chemical approaches based on ketal precursors (1b–d) of
DHAP have been developed.8 These methods were
allowed to obtain DHAP in five or six steps with an
overall yield of 48% starting from acetone8a and 56%
starting from the expensive 1,3-dibromoacetone.8b
The most common syntheses of DHAP are targeted
towards the preparation of the phosphorylated DHAP
dimer 2, a stock material, which was first described by
Colbran et al.9 This original method has been optimized
by modifications of the phosphorylation step of the pro-
tected DHAP dimer. In the conditions reported by
Effenberger and Straub,10 DHAP was obtained in only
three steps with an overall yield of 34%. Two improved
procedures have been developed by Wong and co-work-
ers and led to DHAP in four steps with 55%11 and 61%
yield, respectively.12 Recently, labelled DHAP was
Currently, several enzymatic and chemical syntheses
of DHAP have been reported in the literature.4–13 The
easiest method is the in situ formation of DHAP from
fructose-1,6-biphosphate.4 The fructose 1,6-biphosphate
aldolase catalyzes the reversible condensation of two
Keywords: Dihydroxyacetone phosphate (DHAP);Aldolase.
*
Corresponding author. Tel.: +33 (0)3 90 24 13 49;fax: +33 (0)3 90 24
13 45;e-mail: grosdemange@chimie.u-strasbg.fr
0040-4039/$ - see front matter Ó 2004 Elsevier Ltd. All rights reserved.
doi:10.1016/j.tetlet.2004.08.148

7922
O. Meyer et al. / Tetrahedron Letters 45 (2004) 7921–7923
O
O
HO
OPO(OH)2
R
H
DHAP
FruA
TagA
RhuA
O
FucA
OH
O
OH
OH
O
OH
O
OPO(OH)2
OPO(OH)2
OPO(OH)2
OPO(OH)2
R
R
R
R
OH
OH
OH
OH
Scheme 1. Enzymatic asymmetric aldolization using D-fructose 1,6-biphosphate aldolase = FruA, L-rhamnulose 1-phosphate aldolase = RhuA,
L-fucose 1-phosphate aldolase = FucA, D-tagatose 1,6-biphosphate aldolase = TagA.
2-
Our first attempt was to synthesize the precursor 3 in
two steps starting from the ( )-1-O-benzylglycerol 4, a
commercial protected glycerol. Two methods were
investigated: a selective oxidation of the secondary
alcohol followed by a phosphorylation of the primary
hydroxyl group, or alternatively a selective phosphoryl-
ation of the primary alcohol followed by the oxidation
of the secondary hydroxyl group (Scheme 3).
OPO3
O
O
2
O
R'O
R
OR'
OPO3
2-
EtO
OEt
BnO
OPO(OBn)2
2-O3PO
1a R=OH, R'=OMe
1b R=OAc, R'=OMe
1c R=Br, R'=OH
3
_
_
1d R=OAc, R'= CH2
In order to selectively oxidize the secondary hydroxyl
group, several oxidation conditions were tested like
NBS/acetone or CuBr2/H2O2, but the desired product
was not isolated with satisfactory yields. Consequently,
the selective phosphorylation of the primary hydroxyl
group was investigated. The use of dibenzyl chlorophos-
phate as phosphorylating agent in presence of pyridine
gave the desired phosphate 5. With this method, no
diphosphorylated compound was obtained, but migra-
tion of the phosphate from primary to secondary hydr-
oxyl group was observed, which complicated the
isolation of the desired product. Other phosphorylation
methods like P(OBn)3/I2/pyridine in CH2Cl2 or
HPO(OBn)2/NEt3/CCl4/ultrasounds in toluene were
tested. These conditions were not satisfactory and
yielded a mixture of phosphates on the primary or the
secondary hydroxyl group in both cases. By oxidizing
this mixture with TPAP/NMO, the desired product 3
was easily obtained after flash chromatography with a
24% global yield.
Scheme 2. Known precursors of DHAP.
synthesized via precursor 3 having the alcohol function
and the phosphate ester protected by benzyl groups.13
After a hydrogenation, DHAP was obtained in five steps
with an overall yield of 62%13a or 50%.13b
Many chemical procedures are quite low yielding and
complicated by multistep purification procedures. In
some cases, they involve unstable intermediates and/or
expensive or toxic reagents. We report here a practical
and efficient procedure suitable for a gram-scale synthe-
sis of DHAP from the precursor 3.
2. Results and discussion
Our approach to efficiently synthesize the benzylated
DHAP precursor 3 was based on the chemical modifica-
tions of commercially available starting materials, the
( )-1-O-benzylglycerol 4 and ( )-glycidol 6, with the
same C3 skeleton as DHAP.
In order to optimize the synthesis of precursor 3, we
decided to start from the commercially available ( )-
glycidol 6. The hydroxyl group was phosphorylated,
OP(O)(OBn)2
OH
OH
OH
i
ii
BnO
OH
BnO
OP(O)(OBn)2
O
O
4
5
7
6
iii
O
O
BnO
OH
BnO
OP(O)(OBn)2
3
˚
Scheme 3. Chemical synthesis of precursor 3. (i) (BnO)2P-NEt2, tetrazole, m-CPBA, CH2Cl2 (85%), (ii) BnOH, BF3/Et2O, 4A molecular sieves,
˚
CH2Cl2 (88%), (iii) TPAP, NMO, 3A molecular sieves, CH2Cl2 (82%).

O. Meyer et al. / Tetrahedron Letters 45 (2004) 7921–7923
7923
the epoxy ring selectively opened with benzyl alcohol
following a methodology developed for the synthesis
of phospholipids.14 Finally the secondary alcohol was
oxidized to give the DHAP precursor 3 (Scheme 2).
the DHAP precursor 3, a stable stock material, which
can be can be stored for several months at À18°C and
hydrogenolyzed into DHAP just before its use in
enzyme-catalyzed synthesis. This compound represents
thus a convenient stock material for DHAP.
By using dibenzylchlorophosphate in the presence of
pyridine, the desired phosphate was not obtained.
Decomposition of the starting material and formation
of many by-products were observed. The phosphorami-
date method was therefore used. The epoxy alcohol 6
was treated with dibenzyl-N,N-diethyl phosphoramidite
in the presence of tetrazole to give the corresponding
phosphite, which was readily oxidized with m-CPBA
into phosphate 7.15 The following selective oxirane ring
opening was carried out in CH2Cl2 with boron trifluo-
ride etherate as catalyst and benzyl alcohol as nucleo-
phile.16 Only alcohol 5, resulting from reaction of the
benzyl alcohol on the less hindered side, was obtained.
The next step leading to the protected DHAP was the
oxidation of this compound. The mild oxidation of 5
with TPAP/NMO gave rapidly after silica gel chroma-
tography the precursor 3.17 Using this short synthesis,
the DHAP precursor 3 was obtained with an overall
yield of 61% and was stored for months without notice-
able decomposition. The last step was the deprotection
of all benzyl groups by catalytic hydrogenation. This
was realized at atmospheric pressure in methanol/water
(9/1) with palladium over charcoal (11%). After 1h,
DHAP was obtained in quantitative yield without
side-products. This step required no purification. The
catalyst was simply eliminated by filtration on Celite,
and the filtrate was concentrated under vacuum. The
residue was dissolved in water and the solution was neu-
tralized by addition of 1M sodium hydroxide. DHAP
thus prepared was coupled with butanal using the same
conditions than Schoevaart et al.,18 and the same
aldol, 1,3,4-tri-O-acetyl-5-deoxy-5-ethyl-D-xylulose was
obtained.
Acknowledgements
We thank Mr. J.-D. Sauer for all NMR measurements
and Mr. R. Huber for the MS analysis. This investiga-
tion was supported by a grant to M.R. from the ÔInstitut
Universitaire de FranceÕ. O.M. was a recipient of a grant
`
from the ÔMinistere de la Jeunesse, de lÕEducation
Nationale et de la RechercheÕ.
Supplementary data
Supplementary data associated with this article
can be found, in the online version, at doi:10.1016/
j.tetlet.2004.08.148.
References and notes
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This new synthesis of DHAP occurs in four steps and
with an overall yield of 61% from the ( )-glycidol 6.
This yield ranges among those of the most recent synthe-
ses (56% in BolteÕs synthesis and 62% or 50% in the
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racemic glycidol and non-toxic reagents in a short and
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The short and efficient procedure reported here repre-
sents a completely new route for the preparation of