2926
M. Fonvielle et al. / Bioorg. Med. Chem. Lett. 14 (2004) 2923–2926
while solutions of PGA and PGHz, stored in similar
conditions, remained active for weeks.
aqueous solution was evaporated to dryness, and the
residue dried by coevaporation with pyridine, toluene and
finally dissolved in 30 mL of anhydrous pyridine. 2.5 mL
(
25 mmol) of a commercial solution of glycolonitrile in
So, we think that PGHz and PGA could contribute to
the development of a new class of antimicrobial drugs.
water (55% w/w, Fluka) was similarly coevaporated in the
presence of pyridine, toluene and the syrupy residue
redissolved in the previous solution of b-cyanoethylphos-
phate in pyridine. 100 mmol of DCC were added. The
mixture was stirred 24 h at 60 °C under argon and four
more days at rt. 5 mL of water were added to the black
mixture, and after 1 h the whole reaction medium was
evaporated to dryness. The residue was suspended in
Acknowledgements
We are grateful to Celine Roux for her help and helpful
discussions during the enzymatic tests.
2
00 mL water, and the insoluble DCU filtered. The filtrate
was evaporated, and the black residue extracted 3 times
with 100 mL of boiling dichloromethane. The combined
extracts were evaporated, giving 3 g of crude 1 (and
unreacted glycolonitrile).
References and notes
1
2
3
. Sygusch, J.; Beaudry, D.; Allaire, M. Proc. Natl. Acad.
Sci. U.S.A. 1987, 84, 7846–7850.
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. Singer, M.; Rossmiesse, P.; Cali, B. M.; Liebke, H.; Gross,
C. A. J. Bacteriol. 1991, 173, 6242–6248.
The syrup was redissolved in 90 mL of 1 M cyclohexyl-
amine in water and left 4 h at 0 °C and 1 h at rt. After
evaporation, the residue was extracted twice with dichloro-
methane, redissolved in water and the aqueous solution
again extracted twice with DCM. The aqueous phase
was finally evaporated to give 2.1 g (25%) of crude
dicyclohexylammonium salt of phosphoglycolonitrile (2),
recrystallised from hot ethanol.
Compound 2 was quantitatively converted to the amid-
oxyme 3 by treatment with 2equiv of aqueous hydroxyl-
amine at rt during 6 h. After complete evaporation of the
reaction medium, 3 was recrystallised from hot ethanol.
4
. B o€ ck, A.; Neidhart, F. C. J. Bacteriol. 1966, 92, 470–476.
5. Wehmeier, U. F. FEMSMicrobiol. Lett. 2001, 197, 53–58.
6. Lewis, D. J.; Lowe, G. J. Chem. Soc., Chem. Commun.
1973, 713–715.
7
. Gefflaut, T.; Blonski, C.; Peri ꢀe , J.; Willson, M. Prog.
Biophys. Mol. Biol. 1995, 63, 301–340.
8
9
. Collins, K. D. J. Biol. Chem. 1974, 249, 136–142.
. (a) Miller, T. A.; Witter, D. J.; Belvedere, S. J. Med. Chem.
Selected analytical data:
1
Compound 2: H NMR (D
2
O) d 4.3 (2H, d, 10 Hz) (d 2.9:
2003, 46, 5097–5116; (b) Woessner, J. F., Jr.; Annals,
N. Y. Acad. Sci. 1999, 878, 388–403.
2H, m; d 1.3–1.9: 10H, m; d 0.8–1.3, 10H, m: CHA).
C NMR (D O) d 119.00, 119.08, 61.60, 61.65 (51.1, 31.2,
2
13
1
1
1
0. Nicolaides, D. N.; Varella, E. A. In Chemistry of Acids
Derivatives; Patai, S., Ed.; Wiley: Chichester, 1992; Vol. 2,
pp 875–966.
1. (a) Issa, R. M.; El Shazly, M. F.; Iskander, M. F. Z.
Anorg. Allg. Chem. 1967, 354, 90–97; (b) Supp, G. R. Anal.
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2. (a) Auge, F.; Hornebeck, W.; Decarme, M.; Laronze, J.-Y.
Bioorg. Med. Chem. Lett. 2003, 13, 1783–1786; (b) Artico,
M.; Corelli, F.; Massa, S.; Stefancich, G.; Avigliano, L.;
benafi, O.; Marcozzi, G.; sabatini, S.; Mondovi, B. J. Med.
Chem. 1988, 31, 802–806; (c) Coburn, S. P.; Schalten-
brand, W. E. Biochem. J. 1978, 171, 485–488; (d) Dorn, C.
P.; Zimmerman, M.; Yang, S. S.; Yurewicz, E. C.; Ashe,
B. M.; Frankshun, R.; Jones, H. J. Med. Chem. 1977, 20,
25.2, 24.7: CHA).
1
Compound 3: H NMR (D
2
O) d 4.0 (2H, d, 6 Hz) (d 2.9:
2H, m; d 1.3–1.9: 10H, m; d 0.8–1.3, 10H, m: CHA).
1
3
2
C NMR (D O) d 156.1, 156.2, 61.58, 61.54 (51.1, 31.2,
25.16, 24.7: CHA).
þ
HRMS (EI, M ) (tris-trimethylsilyl derivative): calcd for
11 31 2 5 2
C H N O PSi : 386.1278; found: 386.1272.
16. Enzymes: Rabbit muscle and B. subtilis FDP-aldolases
(Fluka) had specific activities of 13 and 30 U/g, respec-
tively. Aldolase from yeast was partly purified according
to Ref. 17 after disruption of the cells in a French press.
Specific activity: 5.5 U/mg.
Enzymatic assays: DHAP formed by cleavage of FBP by
aldolase was estimated by measuring spectrophotometri-
cally (340 nm) the consumption of NADH in a coupled
system employing a 300-fold excess of glycerophosphate
dehydrogenase (GDH). Incubation medium: Class I
aldolase: triethanolamine–HCl buffer pH 7.6, 0.1 M,
EDTA 1 mM; Class II aldolase: glycylglycine buffer
pH 7.4, 0.1 M, 2-mercaptoethanol 0.01 M (omitted for
B. subtilis aldolase).
1464–1468.
1
3. (a) Ganem, B.; Dong, Y.; Zheng, Y. F.; Prestwich, G. D.
J. Org. Chem. 1999, 64, 5441–5446; (b) Ganem, B. Acc.
Chem. Res. 1996, 29, 340–347.
1
1
4. Weber, P.; Fonvielle, M.; Therisod, M. Tetrahedron Lett.
2003, 44, 9047–9049.
5. Synthesis of PGA (3): A suspension of baryum b-cyano-
ethylphosphate (30 mmol) in water was shaken with
17. Rutter, W. J.; Hunsley, J. R. Methods Enzymol. 1966, 9,
480–486.
þ
Dowex 50 (H ) until all organic material dissolved. The