1
084
Vol. 50, No. 8
ϩ
with DCA (25.8 g, 200 mmol) for 2—10 h in concentrated hydrochloric acid,
in a similar manner to the above.
overnight at 5 °C. The precipitated L-Met·Me iodide was collected by fil-
tration, washed with methanol, and dried; yield 12.9 g (37.0%); mp 155—
2
0
20
Ϫ1
(
4S)-TDC·HCl obtained by reacting for 2 h: Yield 1.99 g (17.5%); [a]D
159 °C (decomp.); [a]D ϩ17.0° (cϭ1.00, water). IR (KBr) cm : 3021,
ϩ6.7° (cϭ1.00, water). (4S)-TDC·HCl obtained by reacting for 4 h: Yield 2996, 2907, 2826, 1616, 1567, 1542, 1438, 1410, 1372, 1342, 1328, 1274,
2
0
1
2
.86 g (25.1%); [a] ϩ6.7° (cϭ1.00, water). (4S)-TDC·HCl obtained by 1050, 992, 871, 782, 762, 547, 441. H-NMR (D O) d: 3.90 (1H, dd, Jϭ6.4,
2
20
ϩ
D
reacting for 5 h: Yield 4.12 g (36.1%); [a]D ϩ6.7° (cϭ1.00, water). (4S)-
6.9 Hz, 3-H), 3.60—3.39 (2H, m, 2-H), 2.98 (6H, s, –S (CH ) ), 2.38 (2H,
3 2
2
0
13
TDC·HCl obtained by reacting for 7 h: Yield 4.80 g (42.1%); [a] ϩ6.8° dd, Jϭ7.1, 15.0 Hz, 1-H). C-NMR (D O) d: 174.9 (–COOH), 56.9 (3-C),
D
2
ϩ
(
cϭ1.00, water). (4S)-TDC·HCl obtained by reacting for 8 h: Yield 4.53 g 55.3 (1-C), 42.0 (2-C), 27.6 (–S (CH ) ). Anal. Calcd for C H INO S: C,
3 2 6 14 2
20
(
39.7%); [a] ϩ6.9° (cϭ1.00, water). (4S)-TDC·HCl obtained by reacting 24.75; H, 4.85; N, 4.81%. Found: C, 24.63; H, 4.89; N, 4.83%.
D
2
0
for 9 h: Yield 4.03 g (35.4%); [a]D ϩ6.9° (cϭ1.00, water). (4S)-TDC·HCl
General Procedure for Syntheses of Four Stereoisomers of Cystathio-
obtained by reacting for 10 h: Yield 3.97 g (34.8%); [a] ϩ6.8° (cϭ1.00, nine To a solution of L-Hcy (0.676 g, 5.00 mmol) in 5 mol dm aqueous
2
0
Ϫ3
D
3
water).
sodium hydroxide (4 cm ) was added (S)-ACP·HCl (0.800 g, 5.00 mmol),
The spectra and analytical data of (4S)-TDC·HCl obtained by reaction stirring for 1 d at room temperature. After allowing to stand the solution
Ϫ1
Ϫ3
3
using 200 mmol of DCA for 6 h: IR (KBr) cm : 2936, 2795, 2578, 2455, overnight, 5 mol dm hydrochloric acid (4 cm ) was added to the solution.
3
1
1
763, 1741, 1637, 1529, 1426, 1390, 1350, 1290, 1266, 1228, 1191, 1092, After evaporation in vacuo, followed by addition of 20 cm of methanol to
055, 1041, 1006, 995, 932, 909, 795, 762, 576. H-NMR (D O) d: 5.23 the residue, sodium chloride was removed by filtration. The filtrate was ad-
1H, s, 2-H), 4.11 (1H, dd, Jϭ2.8, 12.8 Hz, 4-H), 3.26—3.15 (1H, m, 5-H), justed with triethylamine to pH 6 and crude (2S,2ЈS)-CYT was collected by
1
2
(
3
2
0
.05—2.97 (1H, m, 5-H), 2.73—2.64 (1H, m, 6-H), 2.13—1.97 (1H, m, 6-
filtration, washed with ethanol, and dried; yield 0.858 g; [a] ϩ21.2°
D
Ϫ3 3
13
H). C-NMR (D O) d: 172.9 (4-C-COOH), 170.3 (2-C-COOH), 61.3 (2-C), (cϭ1.00, 1 mol dm HCl). After dissolving the (2S,2ЈS)-CYT in 150 cm of
2
6
0.3 (4-C), 29.7 (5-C), 29.4 (6-C). Anal. Calcd for C H ClNO S: C, 31.65;
water under refluxing, followed by standing the solution for 5 d at 5 °C, the
H, 4.43; N, 6.15%. Found: C, 31.43; H, 4.25; N, 6.14%. The H- and C- precipitated (2S,2ЈS)-CYT was collected by filtration, and dried.
6
10
4
1
13
20
NMR spectra of the (4S)-TDC·HCl obtained under other conditions were
(2S,2ЈS)-CYT: Yield 0.493 g (44.4%); mp 285—288 °C (decomp.); [a]
D
Ϫ3 26 Ϫ3
virtually identical to the above spectra.
(
ϩ25.6° (cϭ1.00, 1 mol dm HCl) (lit., [a] ϩ25.3° (cϭ1, 1 mol dm
D
6) 21 Ϫ3 8) Ϫ1
4R)-TDC·HCl was synthesized by reacting D-Met (7.46 g, 50.0 mmol)
with DCA (25.8 g, 200 mmol) for 6 h in concentrated hydrochloric acid, in a 2715, 1651, 1626, 1581, 1519, 1416, 1404, 1362, 1343, 1312, 1277, 1214,
similar manner to that described for (4S)-TDC·HCl.
4R)-TDC·HCl: Yield 4.73 g (41.5%); mp 176—179 °C (decomp.); [a]
Ϫ6.5° (cϭ1.00, water). The IR, H-, and C-NMR spectra were virtually Jϭ6.4 Hz, 2-H), 3.21 (1H, dd, Jϭ4.6, 14.8 Hz, 1Ј-H), 3.10 (1H, dd, Jϭ7.1,
identical to those of (4S)-TDC·HCl. Found: C, 31.71; H,4.35; N, 6.14%.
Synthesis of (4S)-1,3-Thiazane-2,4-dicarboxylic Acid Hydrochloride NMR (1 mol dm DCl) d: 174.1 (–COOH), 173.0 (–COOH), 54.8 (2Ј-C),
from L-Homocysteine A mixture of L-Hcy (0.676 g, 5.00 mmol) and
54.1 (2-C), 33.6 (1Ј-C), 32.0 (3-C), 29.7 (4-C). Anal. Calcd for C H N O S:
HCl), [a]D ϩ23.5° (cϭ1, 1 mol dm HCl) ). IR (KBr) cm : 3000, 2951,
1
1166, 1102, 1074, 920, 885, 846, 782, 757, 718, 699, 689, 554. H-NMR
(1 mol dm DCl) d: 4.36 (1H, dd, Jϭ4.5, 7.1 Hz, 2Ј-H), 4.26 (1H, t,
2
0
Ϫ3
(
D
1
13
13
14.9 Hz, 1Ј-H), 2.82 (2H, t, Jϭ7.3 Hz, 4-H), 2.41—2.15 (2H, m, 3-H). C-
Ϫ3
7
14
2
4
3
GLA·H O (0.460 g, 5.00 mmol) in 10 cm of acetic acid was stirred for 2 h C, 37.83; H, 6.35; N, 12.60%. Found: C, 37.68; H, 6.16; N, 12.54%.
2
at 30 °C. After evaporation in vacuo at 50 °C, followed by dissolving the
The following analogs, (2S,2ЈR)-, (2R,2ЈS)-, and (2R,2ЈR)-CYT were pre-
residue in 1 mol dm hydrochloric acid (10 cm ), the solution was further pared from (R)-ACP·HCl and L-Hcy, (S)-ACP·HCl and D-Hcy, and (R)-
Ϫ3
3
evaporated in vacuo to obtain (2S)-TDC·HCl as the residue. (2S)-TDC·HCl
ACP·HCl and D-Hcy, respectively, in a similar manner to (2S,2ЈS)-CYT.
(2S,2ЈR)-CYT: Yield 0.403 g (36.3%); mp 282—283 °C (decomp.) (lit.,
3
8)
was washed by stirring in 50 cm of THF for 30 min at room temperature,
2
0
20
Ϫ3
24
collected by filtration, and dried; yield 0.674 g (59.1%); [a] ϩ5.8° over 300 °C); [a] ϩ24.5° (cϭ1.00, 1 mol dm HCl) (lit., [a]D ϩ23.5°
D
D
1
Ϫ3
8)
24
Ϫ3
9)
Ϫ1
(
cϭ1.00, water). H-NMR (D O) d: 5.39 (s, 2-H), 5.23 (1H, s, 2-H), 4.7— (1 mol dm HCl), [a]D ϩ23.7° (1 mol dm HCl) ). IR (KBr) cm
:
2
4
3
2
.6 (m, 4-H), 4.11 (1H, dd, Jϭ2.8, 12.8 Hz, 4-H), 3.26—3.15 (1H, m, 5-H), 3037, 2657, 1632, 1545, 1423, 1394, 1344, 1314, 1217, 1155, 1072, 882,
.05—2.97 (1H, m, 5-H), 2.73—2.64 (1H, m, 6-H), 2.6—2.5 (m, 6-H), 842, 751, 586, 552, 528. H-NMR (1 mol dm DCl) d: 4.34 (1H, dd, Jϭ4.6,
.3—2.4 (m, 6-H), 2.13—1.97 (1H, m, 6-H). The IR spectrum was identical 7.0 Hz, 2Ј-H), 4.24 (1H, t, Jϭ6.4 Hz, 2-H), 3.27 (1H, dd, Jϭ4.6, 15.2 Hz, 1Ј-
1
Ϫ3
to that of (4S)-TDC·HCl from L-Met.
H), 3.16 (1H, dd, Jϭ7.1, 15.0 Hz, 1Ј-H), 2.82 (2H, t, Jϭ7.4 Hz, 4-H), 2.41—
1
3
Ϫ3
Preparation of L- and D-Homocysteine (4S)- or (4R)-TDC·HCl 2.18 (2H, m, 3-H). C-NMR (1 mol dm DCl) d: 173.9 (–COOH), 172.8
4.55 g, 20.0 mmol) was dissolved in 100 cm of ethanol by adjusting with (–COOH), 54.8 (2Ј-C), 54.1 (2-C), 33.5 (1Ј-C), 31.9 (3-C), 29.7 (4-C). Anal.
3
(
3
Ϫ3
triethylamine to pH 7. To the solution was added 20 cm of 0.5 mol dm
ethanolic hydroxylamine hydrochloride under reflux (78 °C) and then the
Found: C, 37.47; H, 6.25; N, 12.47%.
8)
(2R,2ЈS)-CYT: Yield 0.391 g (35.2%); mp 281—284 °C (decomp.) (lit.,
2
0
Ϫ3
23
D
mixture was immediately adjusted with triethylamine to pH 6—7. After re- over 300 °C); [a] Ϫ24.7° (cϭ1.00, 1 mol dm HCl) (lit., [a] Ϫ21°
D
Ϫ3
Ϫ3
5)
21
Ϫ3
8)
fluxing the mixture for 25 min, 0.5 mol dm ethanolic hydroxylamine hy- (cϭ1, 1 mol dm HCl), [a]D Ϫ23.0° (cϭ1, 1 mol dm HCl) ). The IR,
3
1
13
drochloride (20 cm ) was added to the mixture and then the mixture was ad-
H-, and C-NMR spectra were virtually identical to those of (2S,2ЈR)-CYT.
justed with triethylamine to pH 6—7. After further refluxing the mixture for
Found: C, 37.45; H, 6.18; N, 12.53%.
2
0
1
h, followed by standing overnight at room temperature, the precipitated L-
(2R,2ЈR)-CYT: Yield 0.360 g (32.4%); mp 284—288 °C (decomp.); [a]
D
Ϫ3
21
Ϫ3
or D-Hcy was collected by filtration, washed with methanol, and dried.
Ϫ25.5° (cϭ1.00, 1 mol dm HCl) (lit., [a] Ϫ24.5° (cϭ1%, 1 mol dm
D
2
0
7)
21
Ϫ3
8)
1
13
L-Hcy: Yield 2.20 g (81.5%); mp 247—249 °C (decomp.); [a] ϩ27.2° HCl), [a]D Ϫ22° (cϭ1, 1 mol dm HCl) ). The IR, H-, and C-NMR
D
Ϫ3
16)
20
Ϫ3
(
(
1
cϭ1.00, 1 mol dm HCl) (lit., [a]D ϩ26.8° (cϭ1, 1 mol dm HCl)). IR spectra were virtually identical to those of (2S,2ЈS)-CYT. Found: C, 37.63;
Ϫ1
KBr) cm : 3156, 2940, 1618, 1586, 1508, 1406, 1347, 1319, 1272, 1250, H, 6.16; N, 12.50%.
1
190, 1157, 966, 838, 754, 700, 654, 542. H-NMR (D O) d: 3.89 (1H, dd,
2
Jϭ5.9, 7.0 Hz, 2-H), 2.73—2.56 (2H, m, 4-H), 2.25—2.07 (2H, m, 3-H).
Acknowledgments This research was financially supported by the Kan-
13
C-NMR (D O) d: 176.7 (–COOH), 56.1 (2-C), 37.2 (3-C), 22.4 (4-C). sai University Grant-in-Aid for the Faculty Joint Research Program, 2002.
2
Anal. Calcd for C H NO S: C, 35.54; H, 6.71; N, 10.36%. Found: C, 35.56;
4
9
2
H, 6.41; N, 10.29%.
References and Notes
1) Finkelstein J. D., Martin J. J., Inter. J. Biochem. Cell Biol., 32, 385—
389 (2000).
2) Ricci G., Santoro L., Achilli M., Matarese R. M., Nardini M., Cav-
allini D., J. Biol. Chem., 258, 10511—10517 (1983).
3) Yu S., Sugahara K., Zhang J., Ageta T., Kodama H., Fontana M.,
Duprè S., J. Chromatogr. B, 698, 301—307 (1997).
4) Brown G. B., du Vigneaud V., J. Biol. Chem., 137, 611—615 (1941).
5) McHale D., Mamalis P., Green J., J. Chem. Soc., 1960, 2847—2850
(1960).
6) Snow M. L., Dombro R. S., Ressler C., J. Org. Chem., 32, 246—248
(1967).
7) Weiss S., Stekol J. A., J. Am. Chem. Soc., 73, 2497—2499 (1951).
8) Jung G., Ottnad M., Rimpler M., Eur. J. Biochem., 35, 436—444
(1973).
2
0
D-Hcy: Yield 2.27 g (84.1%); mp 248—250 °C (decomp.); [a] Ϫ27.2°
D
Ϫ3
16)
20
Ϫ3
(
cϭ1.00, 1 mol dm HCl) (lit., [a]D Ϫ26.8° (cϭ1, 1 mol dm HCl)).
1 13
The IR, H-, and C-NMR spectra were virtually identical to those of L-Hcy.
Anal. Found: C, 35.46; H, 6.46; N, 10.27%.
Preparation of (S)-(3-Amino-3-carboxypropyl)dimethylsulfonium Io-
dide After adding iodomethane (25.5 g, 180 mmol) and 300 cm of
methanol to a solution of L-Met (17.9 g, 120 mmol) in 600 cm of water, the
solution was stirred for 6 d at room temperature. The solution was evapo-
rated in vacuo to give crude (S)-L-Met·Me iodide as the residue. After re-
fluxing a suspension of L-Met·Me iodide (33.2 g) in 200 cm of methanol
for 30 min, L-Met·Me iodide was collected by filtration and washed with
3
3
ϩ
ϩ
3
ϩ
2
0
methanol; yield 20.2 g; [a] ϩ16.5° (cϭ1.00, water). After dissolving the
D
ϩ
3
obtained L-Met·Me iodide in 120 cm of water, followed by adding
3
4
00 cm of methanol to the solution, the mixture was allowed stand