Preparation of D-cycloserine and 13C-labeled D-cycloserine

Article abstract of DOI:10.3987/COM-12-12553

D-Cycloserine (DCS), a second stage drug for the treatment of tuberculosis, is synthesized in 19.8% overall yield from DL-serine methyl ester. This synthetic route gives both enantiomers of cycloserine via a corrected and improved one pot resolution procedure using D- and L-tartaric acids. The route is used to synthesize a 13C-labeled derivative for use in biological studies.

Full text of DOI:10.3987/COM-12-12553

Supporting Information for:
1
3
Preparation of (D)-Cycloserine and C-labeled (D)-Cycloserine
a
a
a
b
*,a
Nathan C. Thacker, Judit Toth, Judy L. Miska, Raul Barletta, and James M. Takacs
a
Department of Chemistry, University of Nebraska-Lincoln, NE 68588-0304
b
Department of Veterinary and Biomedical Sciences, University of Nebraska-Lincoln, NE 68583

13
Supporting Information for Thacker, Toth, Miska, Barletta, and Takacs: Preparation of (D)-Cycloserine and C-labeled (D)-Cycloserine
Contents
13
Synthesis and Characterization of C-labelled D-Cycloserine....................................................................................................................................... 5
1
H of DL-Serine methyl ester hydrochloride (1)........................................................................................................................................................ 7
1
3
C of DL-Serine methyl ester hydrochloride (1)....................................................................................................................................................... 8
DEPT of DL-Cycloserine methyl ester hydrochloride (1)......................................................................................................................................... 9
HMQC of DL-Cycloserine methyl ester hydrochloride (1)..................................................................................................................................... 10
IR of DL-Serine methyl ester hydrochloride (1)...................................................................................................................................................... 11
1
H of DL-Methyl-3-hydroxy-2-(tritylamino)propanoate (2) ................................................................................................................................... 12
1
3
C of DL-Methyl-3-hydroxy-2-(tritylamino)propanoate (2) .................................................................................................................................. 13
DEPT of DL-Methyl-3-hydroxy-2-(tritylamino)propanoate (2).............................................................................................................................. 14
HMQC of DL-Methyl-3-hydroxy-2-(tritylamino)propanoate (2)............................................................................................................................ 15
IR of DL-Methyl-3-hydroxy-2-(tritylamino)propanoate (2).................................................................................................................................... 16
1
3
1
H NMR of DL-Methyl-3-hydroxy-2-(tritylamino)propanoate-1- C (11)............................................................................................................. 17
1
1
1
3
13
C NMR of DL-Methyl-3-hydroxy-2-(tritylamino)propanoate-1- C (11)............................................................................................................. 18
H of DL-O-(1,3-dihydro-2,5-dioxo-1-pyrolidinyl)-N-(triphenylmethyl)-methyl ester (3) .................................................................................... 19
3
C of DL-O-(1,3-dihydro-2,5-dioxo-1-pyrolidinyl)-N-(triphenylmethyl)-methyl ester (3) ................................................................................... 20
DEPT of DL-O-(1,3-dihydro-2,5-dioxo-1-pyrolidinyl)-N-(triphenylmethyl)-methyl ester (3)............................................................................... 21
HMQC of DL-O-(1,3-dihydro-2,5-dioxo-1-pyrolidinyl)-N-(triphenylmethyl)-methyl ester (3)............................................................................. 22
S2

13
Supporting Information for Thacker, Toth, Miska, Barletta, and Takacs: Preparation of (D)-Cycloserine and C-labeled (D)-Cycloserine
IR of DL-O-(1,3-dihydro-2,5-dioxo-1-pyrolidinyl)-N-(triphenylmethyl)-methyl ester (3)..................................................................................... 23
1
1
1
13
H NMR of DL-O-(1,3-dihydro-2,5-dioxo-1-pyrolidinyl)-N-(triphenylmethyl)-1- C methyl ester (12)............................................................... 24
H of DL-Methyl- -aminooxy- -amino(triphenylmethyl)butyrate (4).................................................................................................................... 26
3
C of DL-Methyl- -aminooxy- -amino(triphenylmethyl)butyrate (4)................................................................................................................... 27
DEPT of DL-Methyl- -aminooxy- -amino(triphenylmethyl)butyrate (4).............................................................................................................. 28
HMQC of DL-Methyl- -aminooxy- -amino(triphenylmethyl)butyrate (4)............................................................................................................ 29
IR of DL-Methyl- -aminooxy- -amino(triphenylmethyl)butyrate (4).................................................................................................................... 30
1
1
1
1
13
H NMR of DL-Methyl- -aminooxy- -amino(triphenylmethyl)butyrate-1- C (13).............................................................................................. 31
3
13
C NMR of DL-Methyl- -aminooxy- -amino(triphenylmethyl)butyrate-1- C (13)............................................................................................. 32
H of DL- -aminoxyalanine methyl ester dihydrochloride (5)................................................................................................................................ 33
3
C of DL- -aminoxyalanine methyl ester dihydrochloride (5)............................................................................................................................... 34
DEPT of DL- -aminoxyalanine methyl ester dihydrochloride (5).......................................................................................................................... 35
HMQC of DL- -aminoxyalanine methyl ester dihydrochloride (5)........................................................................................................................ 36
IR of DL- -aminoxyalanine methyl ester dihydrochloride (5)................................................................................................................................ 37
1
H of DL-Cycloserine (6)......................................................................................................................................................................................... 38
1
3
C of DL-Cycloserine (6)........................................................................................................................................................................................ 39
DEPT of DL-Cycloserine (6)................................................................................................................................................................................... 40
HMQC of DL-Cycloserine (6)................................................................................................................................................................................. 41
S3

13
Supporting Information for Thacker, Toth, Miska, Barletta, and Takacs: Preparation of (D)-Cycloserine and C-labeled (D)-Cycloserine
IR of DL-Cycloserine (6)......................................................................................................................................................................................... 42
1
1
1
13
H NMR of DL-Cycloserine-1- C (14)................................................................................................................................................................... 43
H of D-Cycloserine – (L)-Tartaric acid (7) ............................................................................................................................................................ 45
3
C of D-Cycloserine – (L)-Tartaric acid (7) ........................................................................................................................................................... 46
DEPT of D-Cycloserine – (L)-Tartaric acid (7)....................................................................................................................................................... 47
HMQC of D-Cycloserine – (L)-Tartaric acid (7)..................................................................................................................................................... 48
IR of D-Cycloserine – (L)-Tartaric acid (7)............................................................................................................................................................. 49
1
1
1
1
13
H of D-Cycloserine-1- C – (L)-Tartaric acid (15)................................................................................................................................................. 50
3
13
C of D-Cycloserine-1- C – (L)-Tartaric acid (15)................................................................................................................................................ 51
H of D-Cycloserine (9)........................................................................................................................................................................................... 52
3
C of D-Cycloserine (9).......................................................................................................................................................................................... 53
DEPT of D-Cycloserine (9) ..................................................................................................................................................................................... 54
HMQC of D-Cycloserine (9) ................................................................................................................................................................................... 55
IR of D-Cycloserine (9) ........................................................................................................................................................................................... 56
1
13
H of D-Cycloserine-1- C (16) ............................................................................................................................................................................... 57
1
3
13
C NMR of D-Cycloserine-1- C (16) .................................................................................................................................................................... 58
1
3
HRMS of D-Cycloserine (9) vs D-Cycloserine-1- C (16)...................................................................................................................................... 59
S4

13
Supporting Information for Thacker, Toth, Miska, Barletta, and Takacs: Preparation of (D)-Cycloserine and C-labeled (D)-Cycloserine
13
Synthesis and Characterization of C-labelled D-Cycloserine
1
3
13
DL-Methyl-3-hydroxy-2-(tritylamino)propanoate-1- C (11). Commercially available DL-Serine-1- C (250 mg, 2.36 mmol) was
1
3
converted to DL-Serine-1- C methyl ester hydrochloride using the procedure for unlabeled material (1). This material was used
1
3
immediately without subsequent purification. DL-Serine-1- C methyl ester hydrochloride was converted to DL-Methyl-3-hydroxy-2-
1
3
(
tritylamino)propanoate-1- C (11) using the procedure for unlabeled material (2) affording 11 (738 mg, 86% over 2 steps) as a white
1
powder. H NMR (400 MHz, CDCl ) 7.53–7.51 (6H, m), 7.32–7.28 (5H, m), 7.24–7.21 (3H, m), 3.78–3.72 (1H, m), 3.64–3.55 (2H,
3
1
3
m), 3.32 (3H, d, J = 3.8 Hz), 3.05-2.96 (1H, m), 2.37 (1H, t, J = 12.56 Hz); C NMR (100 MHz, CDCl ) 174.0; HRMS (ESI) cald.
For CC H NO (M+Na): 385.1609, found 385.1612 m/z.
3
1
3
2
2
23
3
1
3
DL-O-(1,3-Dihydro-2,5-dioxy-1-pyrolidinyl)-N-(triphenylmethyl)-1- C methyl ester (12). 11 (733 mg, 2.02 mmol) was converted to
1
3
DL-O-(1,3-Dihydro-2,5-dioxy-1-pyrolidinyl)-N-(triphenylmethyl)-1- C methyl ester (12) using the procedure for unlabeled material
1
(
3) affording 12 (660 mg, 71%) as a white powder. H NMR (400 MHz, CDCl ) 7.56–7.53 (6H, m), 7.31–7.26 (6H, m), 7.23–7.18
3
(
3H, m), 4.44–4.40 (1H, m), 4.03–3.98 (1H, m), 3.70–3.63 (1H, m), 3.35 (3H, d, J = 3.8 Hz), 3.10 (1H, dd, J = 10.0 Hz, J = 3.8 Hz),
1
2
1
3
13
2
.69 (4H, s); C NMR (100 MHz, CDCl ) 172.6; HRMS (ESI) cald. For CC H N O (M+Na): 482.1773, found 482.1781 m/z.
3 26 26 2 5
1
3
DL-Methyl- -Aminooxy- -amino(triphenylmethyl)butyrate-1- C (13). 12 (653 mg, 1.42 mmol) was converted to DL-Methyl- -
1
3
Aminooxy- -amino(triphenylmethyl)butyrate-1- C (13) using the procedure for unlabeled material (4) affording 13 (526 mg, 98%)
1
as a colorless oil. H NMR (400 MHz, CDCl ) 7.55–7.52 (6H, m), 7.31–7.26 (6H, m), 7.23–7.18 (3H, m), 5.47 (2H, bs), 4.06–4.01
3
1
3
(
1H, m), 3.77–3.68 (2H, m), 3.24 (3H, d, J = 3.8 Hz), 3.78 (1H, bs); C NMR (100 MHz, CDCl ) 174.3; HRMS (ESI) cald. For
CC H N O (M+Na): 400.1718, found 400.1727 m/z.
3
1
3
2
2
24
2
3
1
3
DL-Cycloserine-1- C (14). 13 (520 mg, 1.38 mmol) was converted to the DiHCl salt using the procedure for unlabeled material (5)
1
3
and used without further purification. DiHCl salt of 13 was then converted to DL-Cycloserine-1- C (14) using the procedure for
1
unlabeled material (6) affording 14 (87.2 mg, 61% over 2 steps) as a white powder. H NMR (400 MHz, d -DMSO) 4.44–4.36 (1H,
6
1
3
13
m), 3.76–3.69 (2H, m); C NMR (100 MHz, d -DMSO) 175.0; HRMS (ESI) cald. For CC H N O (M+H): 104.0541, found
6
2
6
2
2
1
04.0542 m/z.
S5

13
Supporting Information for Thacker, Toth, Miska, Barletta, and Takacs: Preparation of (D)-Cycloserine and C-labeled (D)-Cycloserine
1
3
13
D-Cycloserine-1- C-L-Tartrate (15). 14 (80.3 mg, 0.787 mmol) was converted to D-Cycloserine-1- C-L-Tartrate (15) using the
1
procedure for unlabelled material (8) affording 15 (76.7 mg, 77%) as a white crystalline powder. H NMR (400 MHz, d -DMSO)
6
1
3
13
4
2
.46 (1H, s), 4.05 (2H, s), 3.80 (2H, s); C NMR (100 MHz, d -DMSO) 174.2; HRMS (ESI) cald. For CC H N O (M+H):
6 6 12 2 8
54.0705, found 254.0700 m/z.
1
3
13
D-Cycloserine-1- C (16). 15 (73 mg, 0.288 mmol) was converted to D-Cycloserine-1- C (16) using the procedure for unlabelled
1
material (9) affording 16 (16.9 mg, 57%) as a white powder. H NMR (400 MHz, d -DMSO) 4.43–4.40 (1H, m), 3.76–3.71 (2H,
6
1
3
13
m), 3.60–3.10 (3H, bs); C NMR (100 MHz, d -DMSO) 175.0; HRMS (ESI) cald. For CC H N O (M+H): 104.0541, found
6
2
6
2
2
1
04.0543 m/z.
S6

13
Supporting Information for Thacker, Toth, Miska, Barletta, and Takacs: Preparation of (D)-Cycloserine and C-labeled (D)-Cycloserine
1
H of DL-Serine methyl ester hydrochloride (1)
S7

13
Supporting Information for Thacker, Toth, Miska, Barletta, and Takacs: Preparation of (D)-Cycloserine and C-labeled (D)-Cycloserine
1
3
C of DL-Serine methyl ester hydrochloride (1)
S8

13
Supporting Information for Thacker, Toth, Miska, Barletta, and Takacs: Preparation of (D)-Cycloserine and C-labeled (D)-Cycloserine
DEPT of DL-Cycloserine methyl ester hydrochloride (1)
S9

13
Supporting Information for Thacker, Toth, Miska, Barletta, and Takacs: Preparation of (D)-Cycloserine and C-labeled (D)-Cycloserine
HMQC of DL-Cycloserine methyl ester hydrochloride (1)
S10

13
Supporting Information for Thacker, Toth, Miska, Barletta, and Takacs: Preparation of (D)-Cycloserine and C-labeled (D)-Cycloserine
IR of DL-Serine methyl ester hydrochloride (1)
S11

13
Supporting Information for Thacker, Toth, Miska, Barletta, and Takacs: Preparation of (D)-Cycloserine and C-labeled (D)-Cycloserine
1
H of DL-Methyl-3-hydroxy-2-(tritylamino)propanoate (2)
S12

13
Supporting Information for Thacker, Toth, Miska, Barletta, and Takacs: Preparation of (D)-Cycloserine and C-labeled (D)-Cycloserine
1
3
C of DL-Methyl-3-hydroxy-2-(tritylamino)propanoate (2)
S13

13
Supporting Information for Thacker, Toth, Miska, Barletta, and Takacs: Preparation of (D)-Cycloserine and C-labeled (D)-Cycloserine
DEPT of DL-Methyl-3-hydroxy-2-(tritylamino)propanoate (2)
S14

13
Supporting Information for Thacker, Toth, Miska, Barletta, and Takacs: Preparation of (D)-Cycloserine and C-labeled (D)-Cycloserine
HMQC of DL-Methyl-3-hydroxy-2-(tritylamino)propanoate (2)
S15

13
Supporting Information for Thacker, Toth, Miska, Barletta, and Takacs: Preparation of (D)-Cycloserine and C-labeled (D)-Cycloserine
IR of DL-Methyl-3-hydroxy-2-(tritylamino)propanoate (2)
S16

13
Supporting Information for Thacker, Toth, Miska, Barletta, and Takacs: Preparation of (D)-Cycloserine and C-labeled (D)-Cycloserine
13
1
H NMR of DL-Methyl-3-hydroxy-2-(tritylamino)propanoate-1- C (11)
S17

13
Supporting Information for Thacker, Toth, Miska, Barletta, and Takacs: Preparation of (D)-Cycloserine and C-labeled (D)-Cycloserine
1
3
13
C NMR of DL-Methyl-3-hydroxy-2-(tritylamino)propanoate-1- C (11)
S18

13
Supporting Information for Thacker, Toth, Miska, Barletta, and Takacs: Preparation of (D)-Cycloserine and C-labeled (D)-Cycloserine
1
H of DL-O-(1,3-dihydro-2,5-dioxo-1-pyrolidinyl)-N-(triphenylmethyl)-methyl ester (3)
S19

13
Supporting Information for Thacker, Toth, Miska, Barletta, and Takacs: Preparation of (D)-Cycloserine and C-labeled (D)-Cycloserine
1
3
C of DL-O-(1,3-dihydro-2,5-dioxo-1-pyrolidinyl)-N-(triphenylmethyl)-methyl ester (3)
S20

13
Supporting Information for Thacker, Toth, Miska, Barletta, and Takacs: Preparation of (D)-Cycloserine and C-labeled (D)-Cycloserine
DEPT of DL-O-(1,3-dihydro-2,5-dioxo-1-pyrolidinyl)-N-(triphenylmethyl)-methyl ester (3)
S21

13
Supporting Information for Thacker, Toth, Miska, Barletta, and Takacs: Preparation of (D)-Cycloserine and C-labeled (D)-Cycloserine
HMQC of DL-O-(1,3-dihydro-2,5-dioxo-1-pyrolidinyl)-N-(triphenylmethyl)-methyl ester (3)
S22

13
Supporting Information for Thacker, Toth, Miska, Barletta, and Takacs: Preparation of (D)-Cycloserine and C-labeled (D)-Cycloserine
IR of DL-O-(1,3-dihydro-2,5-dioxo-1-pyrolidinyl)-N-(triphenylmethyl)-methyl ester (3)
S23

13
Supporting Information for Thacker, Toth, Miska, Barletta, and Takacs: Preparation of (D)-Cycloserine and C-labeled (D)-Cycloserine
1
13
H NMR of DL-O-(1,3-dihydro-2,5-dioxo-1-pyrolidinyl)-N-(triphenylmethyl)-1- C methyl ester (12)
S24

13
Supporting Information for Thacker, Toth, Miska, Barletta, and Takacs: Preparation of (D)-Cycloserine and C-labeled (D)-Cycloserine
13
13
C NMR of DL-O-(1,3-dihydro-2,5-dioxo-1-pyrolidinyl)-N-(triphenylmethyl)-1- C methyl ester (12)
S25

13
Supporting Information for Thacker, Toth, Miska, Barletta, and Takacs: Preparation of (D)-Cycloserine and C-labeled (D)-Cycloserine
1
H of DL-Methyl- -aminooxy- -amino(triphenylmethyl)butyrate (4)
S26

13
Supporting Information for Thacker, Toth, Miska, Barletta, and Takacs: Preparation of (D)-Cycloserine and C-labeled (D)-Cycloserine
1
3
C of DL-Methyl- -aminooxy- -amino(triphenylmethyl)butyrate (4)
S27

13
Supporting Information for Thacker, Toth, Miska, Barletta, and Takacs: Preparation of (D)-Cycloserine and C-labeled (D)-Cycloserine
DEPT of DL-Methyl- -aminooxy- -amino(triphenylmethyl)butyrate (4)
S28

13
Supporting Information for Thacker, Toth, Miska, Barletta, and Takacs: Preparation of (D)-Cycloserine and C-labeled (D)-Cycloserine
HMQC of DL-Methyl- -aminooxy- -amino(triphenylmethyl)butyrate (4)
S29

13
Supporting Information for Thacker, Toth, Miska, Barletta, and Takacs: Preparation of (D)-Cycloserine and C-labeled (D)-Cycloserine
IR of DL-Methyl- -aminooxy- -amino(triphenylmethyl)butyrate (4)
S30