Synthesis of 2-C-Methyl-D-erythritol 4-phosphate: The first pathway-specific intermediate in the methylerythritol phosphate route to isoprenoids

Article abstract of DOI:10.1021/ol991299x

(matrix presented) 2-C-Methyl-D-erythritol 4-phosphate (4), formed from 1-deoxy-D-xylulose 5-phosphate (3), is the first pathway-specific intermediate in the methylerythritol phosphate route for the biosynthesis of isoprenoid compounds in bacteria, algae, and plant chloroplasts. In this report, 4 was synthesized from 1,2-propanediol (7) in seven steps with an overall yield of 32% and in an enantiomeric excess of 78%.

Full text of DOI:10.1021/ol991299x

ORGANIC
LETTERS
2
000
Vol. 2, No. 2
15-217
Synthesis of 2-C-Methyl-D-erythritol
-Phosphate: The First
2
4
Pathway-Specific Intermediate in the
Methylerythritol Phosphate Route to
Isoprenoids
Andrew T. Koppisch, Brian S. J. Blagg, and C. Dale Poulter*
Department of Chemistry, UniVersity of Utah, Salt Lake City, Utah 84112
poulter@chemistry.chem.utah.edu
Received December 1, 1999
ABSTRACT
2-C-Methyl-D-erythritol 4-phosphate (4), formed from 1-deoxy-D-xylulose 5-phosphate (3), is the first pathway-specific intermediate in the
methylerythritol phosphate route for the biosynthesis of isoprenoid compounds in bacteria, algae, and plant chloroplasts. In this report, 4 was
synthesized from 1,2-propanediol (7) in seven steps with an overall yield of 32% and in an enantiomeric excess of 78%.
Isoprenoid compounds constitute one of the most chemically
diverse families in Nature, with over 30 000 identified
3-phosphate (1) and pyruvate (2) as the primary substrates
for the biogenesis of 6 (Scheme 1). Although the exact
sequence of transformations in the MEP route to IPP has
yet to be established, the first pathway-specific step is an
enzyme-catalyzed rearrangement/reduction of 3 to 2-C-
1
representatives to date. Isopentenyl diphosphate (IPP, 6)
serves as the universal precursor to the members of this
family, whose carbon skeletons are typically synthesized by
condensing IPP with allylic diphosphates. Recent isotopic
6
methyl-D-erythritol 4-phosphate (MEP, 4). Additional label-
2
labeling studies have established that bacteria, plant chlo-
ing studies have established the direct incorporation of
3
4
7
roplasts, and green algae synthesize 6 from 1-deoxy-D-
xylulose 5-phosphate (3). In contrast to the well-established
mevalonate (MVA) pathway operating in most eukaryotes
methylerythritol into isoprenoid compounds, and MEP has
been identified as a critical metabolite in the development
of the parasite responsible for malaria, Plasmodium falci-
5
8
and archaebacteria, this “mevalonate-independent” or me-
parum.
thylerythritol phosphate (MEP) pathway uses glyceraldehyde
Recently, Zenk and co-workers reported the isolation of
a recombinant Escherichia coli enzyme that catalyzes the
conversion of MEP to a 5′-cytidine diphosphate derivative
(5), as well as the incorporation of 5 into the carotenoids of
Capsicum annuum. Evidence exists which indicates that the
gene responsible for this transformation (ygbP) is conserved
(
1) Poulter, C. D.; Rilling, H. C. Biosynthesis of Isoprenoid Compounds;
Porter, J. W., Spurgeon, S. L., Eds.; Wiley: New York, 1981; Vol. 1, pp
62-209.
2) (a) Flesch, G.; Rohmer, M. Eur. J. Biochem. 1988, 175, 405-411.
b) Rohmer, M.; Sutter, B.; Sahm, H. J. J. Chem. Soc., Chem. Commun.
989, 1471-1472.
3) (a) Eisenreich, W.; Menhard, B.; Hylands, P. J.; Zenk, M. H.; Bacher,
1
(
9
(
1
(
A. Proc. Natl. Acad. Sci. U.S.A. 1996, 93, 6431-6436. (b) Lichtenthaler,
(6) (a) Takahashi, S.; Kuzuyama, T.; Watanabe, H.; Seto, H. Proc. Natl.
Acad. Sci. U.S.A. 1998, 95, 9879-9884. (b) Kuzuyama, T.; Shunji, T.;
Watanabe, H.; Seto, H. Tetrahedron Lett. 1998, 39, 4509-4512.
(7) Duvold, T.; Calf, P.; Bravo, J.; Rohmer, M. Tetrahedron Lett. 1997,
38, 6181-6184.
(8) Jomaa, H.; Wiesner, J.; Sanderbrand, S.; Altincicek, B.; Weidemeyer,
C.; Hintz, M.; Turbachova, I.; Eberl, M.; Zeidler, J.; Lichtenthaler, H. K.;
Soldati, D.; Beck, E. Science 1999, 285, 1573-1576.
H. K.; Schwender, J.; Disch, A.; Rohmer, M. FEBS Lett. 1997, 400, 271-
2
74.
(
4) Schwender, J.; Seeman, M.; Lichtenthaler, H. K.; Rohmer, M.
Biochem. J. 1996, 316, 73-80.
5) Bochar, D. A.; Friesen, J. A.; Stauffacher, C. V.; Rodwell, V. W.
(
ComprehensiVe Natural Products Chemistry; Barton, D., Nakanishi, K.,
Eds.; Elsevier: Oxford, 1999; Vol. 2, pp 15-44.
1
0.1021/ol991299x CCC: $19.00 © 2000 American Chemical Society
Published on Web 12/28/1999

Scheme 1
Scheme 2a
a
2 2
Key: (a) TBDMSCl, N-ethyldiisopropylamine, CH Cl ; (b)
TPAP, NMO, CH
bonylmethyl phosphonate, THF, -20 °C; (d) DIBALH, CH
40 °C; (e) dimethyl chlorophosphate, DMAP, CH Cl ; (f)
modified AD-mix â, t-BuOH/H O, NaHCO , 0 °C; (g) TMSBr/
O; HCl.
2
Cl
2
; (c) KH, (2,2,2-trifluoroethyl)methoxycar-
2
Cl
2
,
-
2
2
2
3
H
2
silylated alcohol 812 in high yield. Subsequent oxidation of
1
3
14
8
with TPAP/NMO gave the protected ketone 9.
1
5
Still modifications of the Horner-Emmons reaction were
used to convert 9 to the protected olefin 10 with increased
°
selectivity for the Z isomer. Warming to -40 C accelerated
the reaction without affecting the yield, but E/Z selectivity
was reduced. Z/E isomers were separated by flash column
chromatography at this stage or after installation of the
phosphate moiety (12). The stereochemistry of the double
bond was established by a NOESY experiment, where the
vinylic proton of the Z isomer had a strong cross-peak with
the protons on the methyl group.
within numerous organisms known to contain the MEP
pathway, and the formation of 5 may represent the next
biosynthetic step toward IPP. Additionally, Lange and
Croteau have reported the isolation of a kinase responsible
for catalyzing the conversion of isopentenyl monophosphate
10
Synthesis of 10 from 15 (Scheme 3) using the method of
(IP) to IPP. Incorporation of IP into isoprenoids of pep-
16
Corey and Katzenellenbogen gave the Z isomer exclusively.
permint suggests that it is a viable intermediate in the MEP
route, and the phosphorylation of IP may represent the final
step in the biosynthesis of IPP from MEP. The remaining
transformations in the MEP pathway have yet to be identi-
fied, although both the MVA and MEP pathways converge
17
Propargyl ester 15 was obtained from methyl chloroformate
and the TBDMS-protected propargyl alcohol 14. Although
this approach gives an overall lower yield of 10, it does
1
1
at IPP.
Scheme 3a
Although Rohmer and co-workers have reported the
7
synthesis of 2-C-methyl-D-erythritol, an efficient route to
the 4-phosphate has not been described. We now describe a
synthesis of optically active 2-C-methyl-D-erythritol 4-phos-
phate from 1,2-propanediol in 78% enantiomeric excess and
an overall yield of 32%.
The route to 2-C-methyl-D-erythritol 4-phosphate is out-
lined in Scheme 2. Treatment of 1,2 propanediol (7) with 1
equiv of tert-butyldimethylsilyl chloride afforded the mono-
(9) Rohdich, F.; Wungsintaweekul, J.; Fellermeier, M.; Sagner, S.; Herz,
S.; Kis, K.; Eisenreich, W.; Bacher, A.; Zenk, M. H. Proc. Natl. Acad. Sci.
U.S.A. 1999, 96, 11758-11763.
(
10) Lange, B. M.; Croteau, R. Proc. Natl. Acad. Sci. U.S.A. 1999, 96,
a
Key: (a) n-BuLi, methyl chloroformate, THF, -78 °C; (b)
1
3714-13719.
11) Hahn, F. M.; Baker, J. A.; Poulter, C. D. J. Bacteriol. 1996, 178,
19-624.
(
2
CuBr -DMS, MeLi, THF, -78 °C.
6
216
Org. Lett., Vol. 2, No. 2, 2000

provide a convenient method for specific incorporation of
carbon or hydrogen isotopes into 4 from commercially
available precursors.
triester methyl groups in 13 were removed by treatment with
trimethylsilyl bromide followed by hydrolysis.21 The tert-
butyldimethylsilyl moiety was removed in the same pot by
increasing the acidity of the aqueous solution. The mixture
Olefin 10 was reduced with 2 equiv of DIBALH followed
by reaction with dimethyl chlorophosphate to give 12. The
phosphotriester was then dihydroxylated via the method of
Sharpless to give 13.18 Preliminary experiments indicated
that the phosphotriester moiety imposed several limitations
on use of the standard AD-mix conditions. We found the
basic nature of the commercial AD-mix reagents promoted
the intramolecular migration of the phosphotriester to the
secondary and tertiary hydroxyl groups in 13 and promoted
hydrolysis to form the corresponding triol. In addition, 12
reacted sluggishly. These problems were circumvented by
the use of “modified” AD-mix conditions with buffering by
3
was neutralized with NaHCO and lyophilized, and the solid
residue was chromatographed on cellulose to give MEP.22
The NMR spectra as well as the optical rotation of the
synthetic 4 are consistent with those reported for an
6
b
enzymatically prepared sample. In summary, the synthetic
routes to MEP, which compliment our previous synthesis
2
3
of 1-deoxy-D-xylulose phosphate, offer a straightforward
approach for synthesizing isotopically labeled materials for
use in biosynthetic experiments.
Acknowledgment. This work was supported by the
National Institutes of Health (Grant GM 25521). A.T.K. is
an American Heart Association Western States Affiliate
Predoctorate Fellow.
19
NaHCO
the dihydroxylation upon the addition of a co-oxidant
methanesulfonamide, NMO) to the mixture was minimal
3
. We observed that the usual rate enhancement of
(
and also led to substantial degradation. For this reason the
asymmetric dihydroxylations were conducted in the absence
of co-oxidant. Our modifications gave a 60% yield of 13
from 12 with a 78% enantiomeric excess.20 The phospho-
Supporting Information Available: Complete experi-
mental procedures for the synthesis of compounds 4, 8-13,
and 15. This material is available free of charge via the
Internet at http://pubs.acs.org.
(
12) Yamamoto, K.; Takemae, M. Bull. Chem. Soc. Jpn. 1989, 62, 2111-
2
113.
OL991299X
(
(
13) Griffith, W. P.; Ley, S. V. Aldrichimica Acta 1990, 23, 13-19.
14) Kozikowski, A. P.; Okita, M.; Kobayashi, M.; Floss, H. G. J. Org.
Chem. 1988, 53, 863-869.
(20) Mosher’s esters of racemic and optically active 13 were synthesized
using (dimethylamino)pyridine and 1 equiv of R-methoxy-R-trifluoro-
methylphenylacetyl chloride. Integration of the benzyl signals versus that
(
15) Still, W. C.; Gennari, C. Tetrahedron Lett. 1983, 24, 4405-4408.
(
16) Corey, E. J.; Katzenellenbogen, J. A. J. Am. Chem. Soc. 1969, 91,
19
1
851-1852.
17) Schlessenger, R. H.; Iwanowicz, E. J.; Springer, J. P. Tetrahedron
Lett. 1988, 29, 1489-1492.
of the TBDMS group verified monoesterification. Analysis of the F spectra
(122.38 ppm (s); 122.86 ppm (s)) was used to calculate the reported
enantiomeric excess of the optically active sample. Dale, J. L.; Dull, D. L.;
Mosher, H. S. J. Org. Chem. 1969, 34, 2543-2549.
(
(
18) Kolb, H. C.; VanNieuwenhze, M. S.; Sharpless, K. B. Chem. ReV.
994, 94, 2483-2547.
19) (a) Bennani, Y. L.; Sharpless, K. B. Tetrahedron Lett. 1993, 34,
079-2082. (b)Vanhessche, K. P. M.; Wang, Z.; Sharpless, K. B.
1
(21) Schmidt, A. H. Aldrichimica Acta 1981, 14, 31-38.
(22) Davisson, V. J.; Woodside, A. B.; Poulter, C. D. Methods Enzymol.
1984, 110, 130-144.
(
2
Tetrahedron Lett. 1994, 35, 3469-3472.
(23) Blagg, B. S. J.; Poulter, C. D. J. Org. Chem. 1999, 64, 1508-1511.
Org. Lett., Vol. 2, No. 2, 2000
217