Original and efficient synthesis of D-cycloserine

Article abstract of DOI:10.1002/ardp.200900316

A simple pathway for the preparation of D-cycloserine is presented. The intermediates and D-cycloserine were characterized by FT-IR, ¹H-NMR spectra and elemental analysis. D-Cycloserine can inhibit the growth of Mycobacterium tuberculosis and can be used as a second-line drug for the treatment of tuberculosis, especially for the use in developing countries.

Full text of DOI:10.1002/ardp.200900316

Arch. Pharm. Chem. Life Sci. 2010, 8, 473–475
473
Short Communication
Original and Efficient Synthesis of D-Cycloserine
1
2
1
1
1
Xiaomeng Li , Xia Meng , Hongdong Duan , Lizhen Wang , Shixiao Wang , Yi Zhang , and
1
1
Dawei Qin
1
School of Chemical Engineering, Shandong Institute of Light Industry, Jinan, PR China
School of Light Chemistry and Environmental Engineering, Shandong Institute of Light Industry, Jinan,
PR China
2
A simple pathway for the preparation of D-cycloserine is presented. The intermediates and D-
1
cycloserine were characterized by FT-IR, H-NMR spectra and elemental analysis. D-Cycloserine can
inhibit the growth of Mycobacterium tuberculosis and can be used as a second-line drug for the treatment
of tuberculosis, especially for the use in developing countries.
Keywords: D-Cycloserine / Hydroxylamine hydrochloride / D-Serine / Synthesis / Thionyl chloride
Received: December 24, 2009; Accepted: February 22, 2010
DOI 10.1002/ardp.200900316
Introduction
Results and discussion
D-Cycloserine or D-4-amino-3-isoxazolidinone (Fig. 1a) is
an analogue of the amino acid D-alanine; it inhibits
the enzymes D-alanine racemase and D-alanine synthetase
In this paper, D-cycloserine was obtained by using an original
and efficient method outlined in Scheme 1. D-2-Amino-3-chloro-
propionic acid chloride 1 was prepared by the reaction of 2.5
equiv. excess of thionyl chloride and 1 equiv. of D-serine (Fig. 1b)
in chloroform at room temperature for 2 h and under reflux
for 4 h. This pathway is an original part of our research. Using
this method, the chlorination and chloroformylation reaction
of D-serine can synchronously take place. Meanwhile, the ami-
dogen of D-serine was protected by the dry HCl.
[
1, 2]. The most important property of D-cycloserine is the
inhibition of the growth of Mycobacterium tuberculosis. In
general, it is more effective against Gram-positive than
Gram-negative bacteria [3]. However, it is seldom used
in the management of tuberculosis due to the toxicity
associated with effective dosages. Nevertheless, the world-
wide resurgence of tuberculosis, the emergence of multiply
drug-resistant tuberculosis, and the problematic use of
available drugs required to treat these infections, have
resulted in the application of D-cycloserine as a second-line
drug for the treatment of tuberculosis [4].
D-2-Amino-3-chloropropionohydroxamic acid
2
was
obtained by the reaction of 1.2 equiv. excess of hydroxyl-
amine hydrochloride and 1 equiv. of intermediate 1 with a
yield of 86% for intermediate 2, much higher than in
methods reported previously.
Numerous studies have been focused on the synthesis of
the D-cycloserine. It can be produced by Streptomyces garyphalus
and Streptomyces orchidaceus [5–7]. But this drug can also be
obtained by chemical synthesis [6, 8, 9].
During the experiment, we found that the temperature, the
volume of the water, and the pH were very important for the
last step. If the temperature is low, the reaction speed was very
slow, especially at the end of the reaction. However, if the
temperature is high, not only the hydroxylamine hydrochloride
easily decomposed, but also the intermediate 2 was easy hydro-
lyzed or eliminated from the reaction. Though hydroxylamine
hydrochloride is unstable in water, using water as the reaction
solvent was feasible as can be demonstrated by the character-
izing result of the product. However, the volume of the water
was to be observed critically. Because the hydroxylamine hydro-
chloride is an alkalescent base, the pH of the mixture should be
controlled in the appropriate range. After repeating the exper-
iment, the best reaction conditions are described in this paper.
Our aim was to prepare D-cycloserine by an original
and efficient method. By using this method, the
synthetic route for D-cycloserine was shorter and had a
good yield.
Correspondence: Hongdong Duan, School of Chemical Engineering,
Shandong Institute of Light Industry, Jinan 250353, PR China.
E-mail: hdduan67@gmail.com
Fax: þ86 531 896-31207
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X. Li et al.
Arch. Pharm. Chem. Life Sci. 2010, 8, 473–475
Synthesis
D-2-Amino-3-chloropropionic acid chloride 1
D-Serine (2.10 g, 0.02 mol) was dissolved in chloroform (40 mL)
and dry HCl was added to the mixture for 10 min. Then,
the mixture was cooled to 08C and thionyl chloride (5.95 g,
0
.05 mol) was added dropwise into the mixture within 1 h.
The mixture was stirred at room temperature for 2 h and then
under reflux for 4 h. The reaction mixture was cooled sub-
sequently and the solid was removed by filtration. The solid
was washed with chloroform and dried in vacuo. Then, a white
powder (3.21 g) was obtained. Yield: 90%; m. p.: 128–1308C; IR
ꢀ
1
1
(
(
KBr, cm ): 3445, 2643, 1750, 1518, 1250, 792; H-NMR
400 MHz, O) d: 4.30 (m, 1H, –CH–), 4.11 (m, 1H,
–), 4.01 (m, 1H, J ¼ 12.6 Hz, –CH –) ppm.
NOCl : C, 20.19; H, 3.39; N, 7.85. Found:
C, 20.18; H, 3.41; N, 3.36.
D
2
J ¼ 12.6 Hz, –CH
2
2
Anal. calcd. for C
3
H
6
3
Figure 1. Chemical structure of: (a) D-cycloserine and (b) D-serine.
D-2-Amino-3-chloropropionohydroxamic acid 2
To a cooled solution of hydroxylamine hydrochloride (2.08 g,
0.03 mol) in absolute ethanol (20 mL) 20 mL of 0.91 N sodium
methoxide (0.03 mol) in methanol were added. The mixture
Conclusion
An original and efficient method for the synthesis of D-cyclo-
serine is reported. Our synthesis is a very simple two-step
procedure; all intermediates are stable, readily crystalline
compounds and the yields are very satisfactory.
was cooled to 08C, the sodium chloride was filtered, and
a solution intermediate 1 (3.57 g, 0.02 mol) in absolute
ethanol (20 mL) was added to the filtrate. This solution was
cooled to 08C and 15 mL of 0.91 N sodium methoxide
(
0.03 mol) were added slowly. The solution was concentrated
in vacuo at room temperature to about 45 mL. The water
20 mL) was added to the solution, 40 mL of 0.46 N hydrochloric
(
Experimental
acid (0.02 mol) were added dropwise and the product was
crystallized. The product was filtered and dried in vacuo, then
2
.94 g solid was obtained. Yield: 84%; m. p.: 191–1938C; IR (KBr,
Material and physical measurements
ꢀ
1
1
cm ): 3445 2643, 1750, 1540, 1518, 1250, 792; H-NMR
400 MHz, D
O) d: 4.30 (d, 1H, –CH–), 4.11 (d, 1H, J ¼ 12.6 Hz,
CH –) ppm. Anal. calcd.
–), 4.01 (s, 1H, J ¼ 12.6 Hz, –CH
for C Cl : C, 20.59; H, 4.61; N, 16.01. Found: C, 20.56;
H, 4.62; N, 16.00.
All reagents were obtained from commercial suppliers and were
used without further purification. Solvents used for the reactions
were purified and dried by conventional methods.
Melting points were taken on a SGW X-4 melting point appa-
1
ratus (Jinghe, Shanghai, China). H-NMR spectra was obtained at
(
–
2
2
2
3
H
8
N
2
O
2
2
400 MHz using a Bruker Avance 400 NMR (Bruker Bioscience,
USA) with tetramethylsilane (TMS, d ¼ 0 ppm) as internal
standard. The infrared spectra were determined on a Bruker
Tensor 27 FT-IR spectrometer (Bruker, Ettlingen, Germany) using
KBr pellets. Elemental analysis was performed by Heraeus Rapid
Analyser (Heraeus, Germany).
D-Cycloserine or D-4-amino-3-isoxazolidone 3
A solution of 3.5 g (0.02 mol) of intermediate 2 in 20 mL of
water was stirred and cooled. The mixture’s pH was adjusted
to 11 by an aqueous sodium hydroxide solution (30% wt).
The mixture was stirred at 258C for 2 h. Then 50 mL of 1:1
Scheme 1. Synthesis of D-cycloserine.
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Arch. Pharm. Chem. Life Sci. 2010, 8, 473–475
Synthesis of D-Cycloserine
475
ethanol/isopropyl alcohol was added. The precipitated salts
were filtered and the filtrate was cooled to 58C. To the cold
well-stirred solution sufficient glacial acetic acid was added
dropwise over a 35-min period to bring the alcoholic solution
to pH 6.0. The crystalline precipitate was filtered, washed twice
with l:l ethanol/isopropyl alcohol and twice with ether. Then,
References
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1
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ꢀ1
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The authors have declared no conflict of interest.
ß 2010 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim
www.archpharm.com