Design, synthesis, in vitro evaluation and apoptosis-induction study of a new serie of

2,6-diketopiperazines as antitumor agents on the MDA-MB-231 cell line.

Flor Paulina Garrido-González,[a] Teresa Mancilla-Percino,[a]* Elvia Mera-Jiménez,[b] and

Cynthia Raquel Trejo-Muñoz[b]
[a]Departamento de Química, Centro de Investigación y de Estudios Avanzados del Instituto
Politécnico Nacional, Ciudad de México, México.
[b] Sección de Estudios de Posgrado e Investigación, Escuela Superior de Medicina del
Instituto Politécnico Nacional, Plan de San Luis y Díaz Miron, C.P. 11340 Ciudad de México,
México.

* Address correspondence to:

Teresa Mancilla Percino Ph.D.
Departamento de Química.
Centro de Investigación y de Estudios Avanzados del Instituto Politécnico Nacional.
Avenida Instituto Politécnico Nacional 2508, Colonia Zacatenco, 07360 Ciudad de México,
México.
FAX: 052 (55) 57473389
E-mail: tmancill@cinvestav.mx

Abstract

Keywords:

Introduction

Experimental Section

Chemistry

Biology

General preparation of 2,6-diketopiperazines

Piperazine-2,6-dione (1a). Yield: 40% white solid. mp. 176-180 °C

(decomp.), 199-202 °C. IR υmax (ATR) cm-1: 3294, 2979, 2676, 1686. 1H-NMR (500
MHz, DMSO-d6) δ: 3.04 (1H, br, H-4, NH), 3.36 (4H, s, H-3, H-5, CH2), 10.79 (1H,
br, H-1, CONHCO). 13
C-NMR (125 MHz, DMSO-d6) δ: 48.91 (C-3, C-5,
*CH2CONH), 173.91 (C-2, C-6, CONH). HR-ESI/TOF-MS [M+H]+ (m/z) cal for

1
C4H7N2O2: 115.050204, found: 115.050208.

(3S)-3-methyl-piperazine-2,6-dione (2a). Yield: 73.4% white solid. mp. 125-127

°C. IR υmax (ATR) cm-1: 3291, 3003, 2815, 1690. 1H NMR (400 MHz, DMSO-d6)
δ: 1.21 (3H, d, J= 7 Hz, H-7, CH3), 3.02 (1H, br, H-4, NH), 3.36 (1H, q, J= 7 Hz,
H-3, *CHCH3), 3.40 (2H, s, overlapped with H2O, H-5, CH2NH), 10.77 (1H, br,
H-1, CONHCO). 13C-NMR (100 MHz, DMSO-d6) δ: 16.11 (C-7, CH3), 48.74 (C-
5, CH2NH), 53.12 (C-3, *CHCONH), 173.97 (C-6,*COCH2NH), 175.72 (C-2,
*
COCHNH). HR-ESI/TOF-MS [M+H]+ (m/z) cal for C5H9N2O2: 129.065854, found: 129.065918.

(3R)-3-methyl-piperazine-2,6-dione (2b). Yield: 66% white solid. mp. 126-

127 °C. IR υmax (ATR) cm-1: 3291, 2994, 2813, 1687. 1H NMR (400 MHz,
DMSO-d6) δ: 1.21 (3H, d, J= 7 Hz, H-7, CH3), 3.03 (1H, br, H-4, NH), 3.38
(1H, q, J= 7 H-3, *CHCH3), 3.43 (2H, s, overlapped with H2O, H-5, CH2NH),
10.77 (1H, br, H-1, CONHCO). 13C-NMR (100 MHz, DMSO-d6) δ: 16.11 (C-
7, CH3), 48.74 (C-5, CH2NH), 53.12 (C-3, *CHCONH), 173.97 (C-
*
6,*COCH2NH), 175.72 (C-2, COCHNH). HR-ESI/TOF-MS [M+H]+ (m/z) cal for C5H9N2O2:
129.065854, found: 129.065893.

(3S)-3-isopropyl-piperazine-2,6-dione (3a). Yield: 79% white solid. mp. 117-

118 °C. IR υmax (ATR) cm-1:3289, 3275, 2977, 2963, 2933, 2813, 1681. 1H NMR
(400 MHz, CDCl3): δ: 0.98 (3H, d, J= 6.8 Hz, H-8´, CH3´ ), 1.11 (3H, d, J= 6.8
Hz, H-8, CH3), 1.83 (1H, br, H-4, NH), 2.49 (1H, m, H-7, *CH(CH3)2), 3.29 (1H,
d, J=4.6 Hz, H-3, *CHNH), 3.59 (1H, d, J= 18 Hz, H-5B, CH2NH), 3.79 (1H, d,
J=18 Hz, H-5A, CH2NH), 8.523(1H, br, H-1, CONHCO ). 13C-NMR (100 MHz,
CDCl3) δ: 17.23 (C-8´, CH3´), 19.52 (C-8, CH3), 27.84 (C-7, *CH(CH3)2), 48.27 (C-5, CH2NH), 62.99
(C-3, *CHCONH), 172.25 (C-6,*COCH2NH), 173.19 (C-2, *COCHNH). HR-ESI/TOF-MS [M+H]+
(m/z) cal for C7H13N2O2: 157.097154, found: 157.097270.

2
 (3R)-3-isopropyl-piperazine-2,6-dione (3b). Yield: 70% white solid. mp. 117-119
°C. IR υmax (ATR) cm-1: 3290, 3276, 2978, 2963, 2930, 2808, 1681. 1H NMR (400
MHz, CDCl3) 0.98 (3H, d, J= 6.8 Hz, H-8´, CH3´ ), 1.11 (3H, d, J= 7.0 Hz, H-8,
CH3), 1.83 (1H, br, H-4, NH), 2.51 (1H, m, H-7, *CH(CH3)2), 3.29 (1H, d, J=4.7
Hz, H-3, *CHNH), 3.59 (1H, d, J= 18 Hz, H-5B, CH2NH), 3.81 (1H, d, J=18 Hz,
H-5A, CH2NH), 8.45 (1H, br, H-1, CONHCO ). 13
C-NMR (100 MHz, CDCl3) δ:
17.22 (C-8´, CH3´), 19.53 (C-8, CH3), 27.84 (C-7, *CH(CH3)2), 48.32 (C-5, CH2NH), 63.02 (C-3,
*CHCONH), 172.08 (C-6,*COCH2NH), 173.04 (C-2, *COCHNH). HR-ESI/TOF-MS [M+H]+ (m/z) cal
for C7H13N2O2: 157.097154, found: 157.097427.

(3S)-3-(2-methylpropyl)-piperazine-2,6-dione (4a). Yield: 72% white solid.

mp: 135-138 °C. IR υmax (ATR) cm-1: 3307, 2958, 2871, 2670, 1705, 1683. 1H
NMR (400 MHz, DMSO-d6) δ: 0.86 (3H, d, J= 6.8 Hz, H-9´, CH3´ ), 0.91 (3H,
d, J= 6.8 Hz, H-9, CH3), 1.41 (1H, ddd, J= 4.6, 9.7 y 14 Hz, H-7B,
*CH2CH(CH3)2), 1.59 (1H, ddd, J= 4.4, 9.4 y 13.8 Hz, H-7A,
*CH2CH(CH3)2), 1.79 (1H, m, H-8, *CH(CH3)2), 2.93 (1H, br, H-4, NH), 3.33
(1H, br, H-3, *CHNH), 3.41 (2H, br, H-5, CH2NH), 10.73 (1H, br, H-1,
CONHCO ). 13
C-NMR (100 MHz, DMSO-d6) δ: 21.78 (C9´, CH3´), 23.32 (C-9, CH3), 24.44 (C-8,
*CH(CH3)2), 38.66 (C-7, *CH2CH(CH3)2 ), 47.98 (C-5, CH2NH), 55.42 (C-3, *CHCONH), 173.92 (C-
*
6,*COCH2NH), 175.95 (C-2, COCHNH). HR-ESI/TOF-MS [M+H]+ (m/z) cal for C8H15N2O2:
171.112804, found: 171.112841.

(3R)-3-(2-methylpropyl)-piperazine-2,6-dione (4b). Yield: 71% white solid.

mp: 136-138 °C. IR υmax (ATR) cm-1: 3306, 2959, 2870, 2672, 1707, 1682. 1H
NMR (400 MHz, CDCl3) δ: 0.94 (3H, d, J= 6.4 Hz, H-9´, CH3´ ), 0.98 (3H, d,
J= 6.2 Hz, H-9, CH3), 1.54 (1H, ddd, J= 5.6, 10.3 y 13.8 Hz, H-7B,
*CH2CH(CH3)2), 1.82 (1H, ddd, J= 4.3, 7.4 Hz, H-7A, *CH2CH(CH3)2), 3.19
(1H, m, H-8, *CH(CH3)2) 1.84 (1H, m, H-4, NH), 3.47 (1H, dd, J=4 y 10 Hz,
H-3, *CHNH), 3.58 (1H, d, J= 20 Hz, H-5B, CH2NH), 3.73 (1H, d, J= 18 Hz,
H-5A, CH2NH), 8.83 (1H, br, H-1, CONHCO ). 13C-NMR (100 MHz, CDCl3) δ: 21.26 (C9´, CH3´), 23.23
3
(C-9, CH3), 24.50 (C-8, *CH(CH3)2), 38.47 (C-7, *CH2CH(CH3)2 ), 47.56 (C-5, CH2NH), 55.79 (C-3,
*CHCONH), 172.44 (C-6,*COCH2NH), 174.61 (C-2, *COCHNH). HR-ESI/TOF-MS [M+H]+ (m/z) cal
for C8H15N2O2: 171.112804, found: 171.113078.

(3R)-3-(2-methylpropyl)-piperazine-2,6-dione (4b). Yield: 71% white

solid. mp: 136-138 °C. IR υmax (ATR) cm-1: 3306, 2959, 2870, 2672, 1707,
1682. 1H NMR (500 MHz, CDCl3) δ: 0.91 (3H, d, J= 6.4 Hz, H-9´, CH3´ ),
0.95 (3H, d, J= 6.4 Hz, H-9, CH3), 1.52 (1H, ddd, J= 4.3, 9.7 y 13.7 Hz, H-
7B, *CH2CH(CH3)2), 1.78 (1H, ddd, J= 4.2, 9.5 y 13.5 Hz, H-7A,
*CH2CH(CH3)2), 1.82 (1H, m, H-8, *CH(CH3)2), 2.0 (1H, br, H-4, NH) 3.44
(1H, dd, J=4.3 y 9.8 Hz, H-3, *CHNH), 3.55 (1H, d, J=18 Hz, H-5B,
CH2NH), 3.69 (1H, d, J= 18 Hz, H-5A, CH2NH), 8.89 (1H, br, H-1, CONHCO). 13C-NMR (125 MHz,
CDCl3) δ: 21.32 (C9´, CH3´), 23.23 (C-9, CH3), 24.54 (C-8, *CH(CH3)2), 38.52 (C-7, *CH2CH(CH3)2),
47.61 (C-5, CH2NH), 55.82 (C-3, *CHCONH), 172.59 (C-6,*COCH2NH), 174.76 (C-2, *COCHNH).
HR-ESI/TOF-MS [M+H]+ (m/z) cal for C8H15N2O2: 171.112804, found: 171.113078.

(3S)-3-(1-methylpropyl)-piperazine-2,6-dione (5a). Yield: 62% white

solid. mp:79-80°C. IR υmax (ATR) cm-1: 3282, 2964, 2932, 2876, 2796, 1714,
1680. 1H NMR (500 MHz, CDCl3) δ: 0.91 (3H, t, J= 7.1 Hz, H-10,CHCH3),
1.04 (3H, d, J= 6.9 Hz, H-9, CH2CH3), 1.21 (1H, m, H-8B, *CH2CH3), 1.45
(1H, m, H-8A, *CH2CH3), 1.80 (1H, br, H-4, NH), 2.17 (1H, m, H-7,
CH3CH(CH2CH3)), 3.29 (1H, s, H-3, *CHNH), 3.52 (1H, d, J= 17.8 Hz, H-
13
5B, CH2NH), 3.73 (1H, d, J= 17.8 Hz, H-5A, CH2NH), 8.59 (1H, br, H-1, CONHCO). C-NMR (125
MHz, CDCl3) δ: 11.99 (C-10, CH*CH3), 16.22 (C-9, CH2*CH3), 24.83 (C-8, *CH2CH3), 34.67 (C-7,
CH3*CH(CH2CH3), 48.39 (C-5, CH2NH), 62.78 (C-3, *CHCONH), 172.29 (C-6,*COCH2NH), 173.38
(C-2, *COCHNH). HR-ESI/TOF-MS [M+H]+ (m/z) cal for C8H15N2O2: 171.112804, found: 171.112663.

(3S)-Tetrahydropyrrolo[1,2-a]pyrazine-1,3(2H,4H)-dione (6a). Yield: 79%

white solid. mp: 122-124 °C. IR υmax (ATR) cm-1: 2971, 2914, 2778, 1714, 1689.
1
H NMR (500 MHz, CDCl3) δ 1.88 (2H, quintet, J= 7, H-8, NCH2*CH2CH2),

4
2.22 (2H, m, H-9, NCH2CH2*CH2), 2.73 (1H, dt, J= 7 y 9 Hz, H-7B, N*CH2CH2CH2), 3.39 (1H, t, J= 6.8
Hz, H-3, *CHNH), 3.52 (1H, d, J= 17.5 Hz, H-5B, CH2NH), 3.77 (1H, d, J= 17.3 Hz, H-5A, CH2NH),
13
7.16 (1H, dt, J= 7 y 12.5 Hz, H-7A, N*CH2CH2CH2), 8.97 (1H, br, H-1, CONHCO). C-NMR (125
MHz, CDCl3) δ: 26.10 (C-8, NCH2*CH2CH2), 26.14 (C-9, NCH2CH2*CH2), 52.22 (C-7,
N*CH2CH2CH2), 53.33 (C-5, CH2NH), 62.79 (C-3, *CHCONH), 171.32 (C-6,*COCH2NH), 173.20 (C-2,
*
COCHNH). HR-ESI/TOF-MS [M+H]+ (m/z) cal for C7H11N2O2: 155.081504, found: 155.081517.

(3R)-Tetrahydropyrrolo[1,2-a]pyrazine-1,3(2H,4H)-dione (6b). Yield: 70%

white solid. mp: 121-124 °C. IR υmax (ATR) cm-1: 2971, 2915, 2771, 1715, 1688. 1H
NMR (500 MHz, DMSO-d6) δ 1.69 (2H, quintet, J= 6.3, H-8, NCH2*CH2CH2), 1.93
(2H, m, H-9, NCH2CH2*CH2), 2.54 (1H, dt, J= 15.4 Hz, H-7B, N*CH2CH2CH2),
2.67 (1H, dt, J= 7.9 y 14.6 Hz, H-7A, N*CH2CH2CH2), 3.33 (1H, dd, J= 7.4 y 10.4
Hz, H-3, *CHNH), 3.35 (1H, d, J= 16.9 Hz, H-5B, CH2NH), 3.45 (1H, d, J= 17.3
Hz, H-5A, CH2NH), 10.95 (1H, br, H-1, CONHCO)). 13
C-NMR (125 MHz, DMSO-d6) δ: 22.18 (C-8,
NCH2*CH2CH2), 26.0 (C-9, NCH2CH2*CH2), 52.13 (C-7, N*CH2CH2CH2), 53.55 (C-5, CH2NH), 62.52
(C-3, *CHCONH), 172.49 (C-6,*COCH2NH), 174.24 (C-2, *COCHNH). HR-ESI/TOF-MS [M+H]+
(m/z) cal for C7H11N2O2: 155.081504, found: 155.081563.

(3S)-3-benzylpiperazine-2,6-dione (7a). Yield: 63% white solid. mp: 174-

176 °C. IR υmax (ATR) cm-1: 3733, 3281, 2984, 2718, 1711, 1689. 1H NMR
(500 MHz, DMSO-d6) δ: 2.72 (1H, dd, J= 9.7 y 14.2 Hz, H-7B, *CH2Ph),
2.85 (1H, dt, J=3.3 y 9 Hz, H-4, NH), 3.13 (1H, dd, J= 3.7 y 14.2 Hz, H-7A,
*CH2Ph), 3.32 (1H, dd, J=9.3 y 16.4 H-5B, *CH2NH) ), 3.41 (1H, dd, J= 3.7
y 14.2 Hz, H-5A, *CH2NH)), 3.54 (1H, dt, J= 3.8 y 9.45 Hz, H-3, *CHNH),
7.22 (5H, m, H-9, H-9´, H-10, H-10´ y H-11, Ph), 10.84 (1H, br, H-1, CONHCO). 13C-NMR (125 MHz,
DMSO-d6) δ: 35.70 (C-7, *CH2Ph ), 48.30 (C-5, CH2NH), 58.78 (C-3, *CHCONH), 126.62 (C-11,
aromatic), 128.58 (C-9, aromatic), 129.86 (C-10, aromatic), 139.18 (C-8, aromatic), 173.94 (C-
*
6,*COCH2NH), 174.92 (C-2, COCHNH). HR-ESI/TOF-MS [M+H]+ (m/z) cal for C11H13N2O2:
205.097154, found: 205.097425.

5
 (3R)-3-benzylpiperazine-2,6-dione (7b). Yield: 78% white solid. mp: 174-
176 °C. IR υmax (ATR) cm-1: 3281, 2985, 2722, 1712, 1692. 1H NMR (400
MHz, DMSO-d6) δ: 2.75 (1H, dd, J= 9.8 y 14 Hz, H-7B, *CH2Ph), 2.90 (1H,
dt, J=9.3 y 15.1 Hz, H-4, NH), 3.16 (1H, dd, J= 3.6 y 14 Hz, H-7A,
*CH2Ph), 3.35 (1H, dd, J=5.4 y 17.6 H-5B, *CH2NH), 3.44 (1H, dd, J=5.4 y
17.6 Hz, H-5A, *CH2NH)), 3.57 (1H, dt, J= 3.6 y 9.5 Hz, H-3, *CHNH),
7.24 (5H, m, H-9, H-9´, H-10, H-10´ y H-11, Ph), 10.81 (1H, br, H-1, CONHCO). 13C-NMR (100 MHz,
DMSO-d6) δ: 35.72 (C-7, *CH2Ph), 48.33 (C-5, CH2NH), 58.80 (C-3, *CHCONH), 126.64 (C-11,
aromatic), 128.61 (C-10, aromatic), 129.89 (C-9, aromatic), 139.21 (C-8, aromatic), 173.97 (C-
*
6,*COCH2NH), 174.94 (C-2, COCHNH). HR-ESI/TOF-MS [M+H]+ (m/z) cal for C11H13N2O2:
205.097154, found: 205.097374.

(3S)-3-(4-hydroxybenzyl)piperazine-2,6-dione (8a). Yield: 71%

white solid. mp: 171-174 °C. IR υmax (ATR) cm-1: 3399, 3287, 3044,
2879, 2828, 1721, 1694. 1H NMR (400 MHz, DMSO-d6) δ: 2.68 (1H,
dd, J= 9.3 y 14.1 Hz, H-7B, *CH2Ph), 2.78 (1H, br, H-4, NH), 3.03
(1H, dd, J= 3.4 y 14.3 Hz, H-7A, *CH2Ph), 3.34 (1H, overlapped with
H2O, H-5B, *CH2NH), 3.41 (1H, d, J= 3.4 Hz, H-5A, *CH2NH), 3.47
(2H,d, J= 4.8, H-3, *CHNH), 6.85 (2H, d, J= 8.0 Hz, H-9, H-9A´, aromatic), 7.02 (2H, d, J= 8 Hz, H-
13
10, H-10A´,aromatic ), 9.16 (1H, br, H-12, OH), 10.81 (1H, br, H-1, CONHCO). C-NMR (100 MHz,
DMSO-d6) δ: 34.84 (C-7,*CH2Ph), 48.26 (C-5, CH2NH), 58.93 (C-3, *CHCONH), 115.35 (C-10,
aromatic), 128.88 (C-11, aromatic), 130.72 (C-9, aromatic), 156.18 (C-8, aromatic), 173.83 (C-
*
6,*COCH2NH), 174.83 (C-2, COCHNH). HR-ESI/TOF-MS [M+H]+ (m/z) cal for C11H13N2O3:
221.092068, found: 221.092088.

(3S)-2-(4-((3,5-dioxopiperazin-2-
yl)methyl)phenoxy)acetamide (8a´): Yield: 67% yelow
solid. mp: 192-194 °C. IR υmax (ATR) cm-1: 3396, 3284,
3207, 2993, 2773, 1698, 1660. 1H NMR (500 MHz,

6
DMSO-d6) δ: 2.67 (1H, dd, J= 9.6 y 14.1 Hz, H-7B, *CH2Ph), 2.80 (1H, dt, J= 9.2 y 14.9 Hz H-4, NH),
3.05 (1H, dd, J= 3.6 y 14.1 Hz, H-7A, *CH2Ph), 3.31 (1H, dd, J= 9.2 y 17.7 Hz, H-5B, *CH2NH), 3.40
(1H, dd, J= 5.3 y 17.7 Hz, H-5A, *CH2NH), 3.47 (1H, td, J= 3.6 y 9.6, H-3, *CHNH), 4.34 (2H, s, H-13,
O*CH2CONH2), 7.35 (1H, br, H-15A, CONH2), 7.46 (1H, br, H-15B, CONH2), 6.8 (2H, d, J= 8.5 Hz, H-
10, H-10A´,aromatic ), 7.12 (2H, d, J= 8.6 Hz, H-9, H-9A´, aromatic), 7.46 (1H, br, H-15A, CONH2), 7.35
(1H, br, H-15B, CONH2),10.80 (1H, br, H-1, CONHCO). 13
C-NMR (125 MHz, DMSO-d6) δ: 34.79 (C-
7,*CH2Ph), 48.29 (C-5, CH2NH), 58.88 (C-3, *CHCONH), 67.27 (C-13, O*CH2CONH2), 114.82 (C-10,
aromatic), 130.98 (C-9, aromatic), 131.58 (C-8, aromatic), 156.76 (C-11, aromatic), 170.66 (C-14,
*CONH2), 173.95 (C-6,*COCH2NH), 174.92 (C-2, *COCHNH). HR-ESI/TOF-MS [M+H]+ (m/z) cal for
C13H16N3O2: 278.113532 found 278.113351.

(3R)-3-(4-(benzyloxy)benzyl)piperazine-2,6-dione)
(8b´). Yield: 51% white solid. mp: 181-183 °C. IR υmax
(ATR) cm-1:3282, 2988, 2767, 2719, 1693, 1
H NMR
(400 MHz, DMSO-d6) δ: 2.73 (1H, dd, J= 9.2 y 14.9 Hz,
H-7B, *CH2Ph), 2.83 (1H, dt, J= 9.3 y 14.9 Hz H-4,
NH), 3.09 (1H, dd, J= 3.9 y 14.9 Hz, H-7A, *CH2Ph),
3.36 (1H, dd, J=9, 17.3, H-5B, *CH2NH), 3.44 (1H,
overlapped with H2O, H-5A, *CH2NH), 3.51 (1H, td, J= 3.9 y 9.2 Hz, H-3, *CHNH), 6.85 (2H, d, J= 8.0
Hz, H-9, H-9A´, aromatic), 5.06 (2H, s, H-13, OCH2Ph ), 6.91 (2H, d, J=8.8, H9A y H9A´aromatic), 7.16 (
2H, d, J=8.5, H10A Y H10A´, aromatic), 7.38 (5H, m, H15, H16, H17, aromatic), 10.84 (1H, br, H-1,
CONHCO). 13
C-NMR (100 MHz, DMSO-d6) δ: 34.77 (C-7,*CH2Ph), 48.23 (C-5, CH2NH), 58.82 (C-3,
*CHCONH), 69.59 (C-13, OCH2Ph), 114.88 (C-8, aromatic), 128.88 (C-11, aromatic), 128.07 (C-15,
aromatic), 128.238 (C-17, aromatic), 128.89 (C-10, aromatic), 130.83 (C-9, aromatic), 131.03 (C-16,
aromatic), 137.72 (C-14, aromatic), 157.30 (C-11, aromatic), 173.86 (C-6,*COCH2NH), 174.86 (C-2,
*
COCHNH). HR-ESI/TOF-MS [M+H]+ (m/z) Cal for C18H19N2O3: 311.139019, found: 311.139129.

(3S)-3-(hydroxymethyl)piperazine-2,6-dione (9a). Yield: 78% white solid mp:

158-160 °C. IR υmax (ATR) cm-1: 3302, 3232, 3078, 2746, 1729, 1702. 1H NMR
(500 MHz, DMSO-d6) δ: 3.03 (1H, br, H-4, NH), 3.36 (1H, overlapped with
H2O, H-3, *CHNH ), 3.42 (1H, d, J=17.6, H-5B, *CH2NH), 3.53 (1H, d, J=17.6,

7
H-5A, *CH2NH), 3.65 (1H, d, J=10.5 Hz, H-7A, CH2OH), 3.83 (1H, dd, J=10.2 H-7B, CH2OH), 4.88
(1H, t, J=5.3, H-8, OH), 10.83 (1H, br, H-1, CONHCO). 13
C-NMR (125 MHz, DMSO-d6) δ: 48.45 (C-5,
CH2NH), 59.16 (C-3, *CHCONH), 61.17 (C-7, CH2OH), 174.03 (C-6,*COCH2NH), 174.17 (C-2,
*
COCHNH). HR-ESI/TOF-MS [M+H]+ (m/z) cal for C5H9N2O3: 145.060768, found: 145.060953.

(3R)-3-(hydroxymethyl)piperazine-2,6-dione (9b). Yield: 78% pink solid. mp:

158-160 °C. IR υmax (ATR) cm-1: 3300, 3230, 3086, 2686, 1730, 1702. 1H NMR
(400 MHz, DMSO-d6) δ: 2.93 (1H, br, H-4, NH), 3.06 (1H, m, H-7B, CH2OH),
3.32 (1H, br, H-3, *CHNH ), 3.38 (1H, overlapped with H2O, H-5B, *CH2NH),
3.48 (1H, d, J=17.4, H-5A, *CH2NH), 3.78 (1H, dt, J=5.3 y 10.4 Hz, H-7A,
CH2OH), 4.88 (1H, t, J=5.3, H-8, OH H8= FALTA), 10.79 (1H, br, H-1,
CONHCO). 13C-NMR (100 MHz, DMSO-d6) δ: 48.41 (C-5, CH2NH), 59.12 (C-3,*CHCONH), 61.30 (C-
7, CH2OH), 173.95 (C-6,*COCH2NH), 174.11 (C-2, *COCHNH). HR-ESI/TOF-MS [M+H]+ (m/z) cal
for C5H9N2O3 145.097154, found: 145.060836.

Methyl 6-oxothiomorpholine-3-carboxylate (10a´). Yield: 85% yellow

liquid. IR υmax (ATR) cm-1: 3258, 2912, 1728, 1621. 1H NMR (500 MHz,
CDCl3) δ: 2.94 (1H, dd, J=7.7 y 13.5 Hz, H-8B, CH2SCO), 3.09 (1H, dd,
J=3.9 y 13.4 Hz, H-8A, CH2SCO), 3.30 (1H, d, J=7, H-5A, NHCH2CO ),
3.78 (3H, s, H-1, CO2CH3), 4.37 (1H, dd, J=2.8 y 3.5 Hz, H-3,
*CHCO2CH3), 6.91 (1H, br, H-4, NH). 13
C-NMR (125 MHz, CDCl3) δ: 27.48 (C-8, SCH2), 29.68 (C-5,
CO*CH2NH), 53.23 (C-1, CO2*CH3), 57.00 (C-3, *CHCO2CH3), 166.57 (C-6,*COCH2NH), 169.72 (C-2,
*
CO2CH3). HR-ESI/TOF-MS [M+H]+ (m/z) cal for C6H10NO3S: 176.037591, found 176.037895.

(3R)-((benzylthio)methyl)piperazine-2,6-dione (10aa). Yield: 95%

purple solid. mp: 115-117 °C. IR υmax (ATR) cm-1: 3248, 3164, 3030,
2827, 1695. 1H NMR (400 MHz, CDCl3) δ: 2.20 (1H, br, H-4, NH),
3.06 (1H, m, H-7B, CH2OH), 3.32 (1H, br, H-3, *CHNH ), 3.38
(1H, overlapped with H2O, H-5B, *CH2NH), 3.48 (1H, d, J=17.4, H-
5A, *CH2NH), 3.78 (1H, dt, J=5.3 y 10.4 Hz, H-7A, CH2OH), 4.88

8
(1H, t, J=5.3, H-8, OH H8= FALTA), 10.52 (1H, br, H-1, CONHCO). 13
C-NMR (100 MHz, CDCl3) δ:
48.41 (C-5, CH2NH), 59.12 (C-3,*CHCONH), 61.30 (C-7, CH2OH), 173.95 (C-6,*COCH2NH), 174.11
(C-2, *COCHNH). HR-ESI/TOF-MS [M+H]+ (m/z) cal for

(3S)-3-(hydroxyethyl)piperazine-2,6-dione (11a). Yield: 81% white solid.

mp: 153-154 °C. IR υmax (ATR) cm-1: 1H NMR (400 MHz, DMSO-d6) δ: 1.19
(1H, d, J=13.3 H-9, CH(OH)CH3), 2.82 (1H, br, H-4, NH), 3.15 (1H, s, H-3,
*CHNH), 3.36 (1H, d, J=17.5, H-5B, *CH2NH), 3.56 (1H, d, J=17.5, H-5A,
*CH2NH), 4.27 (1H, br, H-7, *CH2OH), 4.84 (1H, d, J=4.3, H-8, OH), 10.77
(1H, br, H-1, CONHCO). 13
C-NMR (100 MHz, DMSO-d6) δ: 21.16 (C-9,
CH(OH)*CH3), 48.04 (C-5, CH2NH), 62.22 (C-3, *CHCONH), 66.03 (C-7,
CH2OH), 174.14 (C-6,*COCH2NH), 174.75 (C-2, *COCHNH). (m/z) cal for C6H11N2O3:171.1.

(3R)-3-(hydroxyethyl)piperazine-2,6-dione (11b). Yield: 70% white solid.

mp: 152-154 °C. IR υmax (ATR) cm-1: 1H NMR (400 MHz, DMSO-d6) δ: 1.19
(1H, d, J=6.3 H-9, CH(OH)CH3), 2.81 (1H, br, H-4, NH), 3.15 (1H, s, H-3,
*CHNH), 3.38 (1H, dd, J=6.3 y 18 Hz, H-5B, *CH2NH), 3.56 (1H, d, J=17 Hz,
H-5A, *CH2NH), 4.27 (1H, dq, J=2.4 y 4.9 y possible 6.3 Hz o qdd, H-7,
13
*CH2OH), 4.84 (1H, d, J=4.9, H-8, OH), 9.16 (1H, br, H-1, CONHCO). C-
NMR (100 MHz, DMSO-d6) δ: 20.94 (C-9, CH(OH)*CH3), 48.04 (C-5,
CH2NH), 61.96 (C-3, *CHCONH), 65.91 (C-7, CH2OH), 174.13 (C-6,*COCH2NH), 174.75 (C-2,
*
COCHNH). (m/z) cal for C6H11N2O3: 171.1.

(3S)-3-(2-(methylthio)ethyl)piperazine-2,6-dione (12a). Yield: 82% white

solid. mp: 64-66 °C. IR υmax (ATR) cm-1:3281, 3163, 3037, 2949, 2916,
2829, 1686. 1H NMR (400 MHz, CDCl3) δ: 2.13 (3H, s, H-10, CH3S), 2.26
(2H, m o td, H-4,H7B, NH, *CH2CH2SCH3), 2.33 (1H, td, J=4.1, 7.2 Hz, H-
7A, *CH2CH2SCH3), 2.72 (2H, dd, J=5.8,J=12, H-8, CH2*CH2SCH3), 3.61

9
(1H, d, J=18 Hz, H-5B, *CH2NH), 3.67 (1H, dd, J= 4.1 y 8.8 Hz, H-3, *CHNH), 3.77 (1H, d, J=17.8 Hz,
H-5A, *CH2NH), 8.56 (1H, br, H-1, CONHCO). 13
C-NMR (100 MHz, CDCl3) δ: 15.26 (C-10, SCH3),
28.78 (C-7, *CH2CH2SCH3), 30.37 (C-8, CH2*CH2SCH3), 48.10 (C-5, CH2NH), 56.35 (C-3,
*CHCONH), 171.94 (C-6,*COCH2NH), 173.58 (C-2,*COCHNH). HR-ESI/TOF-MS [M+H]+ (m/z) cal
for C7H13N2O2S: 189.069226, found: 189.069304.

(3R)-3-(2-(methylthio)ethyl)piperazine-2,6-dione (12b). Yield: 80% white solid. mp: 65-67 °C. IR υmax
(ATR) cm-1: 3288, 3160, 3045, 2949, 2914, 2878, 2827, 1687. 1H NMR
(400 MHz, CDCl3) δ: 1.80 (1H, br, H-4, NH), 1.93 (1H, dtd, J=5.8, 6.8, 22
Hz, H-7B, *CH2CH2SCH3), 2.13 (3H, s, H-10, CH3S), 2.34 (1H, dtd,
J=4.3, 7.7 y 14.6 Hz, H-7A, *CH2CH2SCH3), 2.71 (1H, dd, J=4.8,J=5.8, H-
8A, CH2*CH2SCH3), 2.72 (1H, dd, J=4.8,J=5.9, H-8B, CH2*CH2SCH3),
3.61 (1H, d, J=18 Hz, H-5B, *CH2NH), 3.66 (1H, dd, J= 4.3 y 8.7 Hz, H-3,
13
*CHNH), 3.77 (1H, d, J=18 Hz, H-5A, *CH2NH), 8.56 (1H, br, H-1, CONHCO). C-NMR (100 MHz,
CDCl3) δ: 15.26 (C-10, SCH3), 28.78 (C-7, *CH2CH2SCH3), 30.37 (C-8, CH2*CH2SCH3), 48.10 (C-5,
CHNH), 56.35 (C-3, *CHCONH), 171.94 (C-6,*COCH2NH), 173.58 (C-2,*COCHNH). HR-ESI/TOF-
MS [M+H]+ (m/z) cal for C7H13N2O2S: 189.069226, found: 189.069392.

(3S)-benzyl-(4-(3,5-dioxopiperazin-2-
yl)butyl)carbamate (13a´). Yield: 64% white solid.
mp: 114-116 °C. IR υmax (ATR) cm-1: 1H NMR (400
MHz, CDCl3) δ: 1.43 (H3, m, H-9,H-7B
*CH2CH2*CH2CH2NH),1.77 (2H, m,
CH2*CH2CH2CH2NH, H-8), 2.98 (1H, m O br, H-7B,
*CH2CH2CH2CH2NH), 3.25 (1H, br, H-4, NH), 3.35
(2H, br, H-10, CH2CH2CH2*CH2NH), 3.41 (1H, br, H-3,*CHNH), 3.74 (1H, s, H-5A, ,*CH2NH), 5.01
(1H, s, H-14, O*CH2Ph ), 7.16 (1H, br, H-11, COONH), 7.31 (5H, m, H-16, H-17, H-18,Ph), 10.75 (1H,
br, H-1, CONHCO), 13
C-NMR (100 MHz, CDCl3) δ: 23.22 (C-8, CH2*CH2CH2CH2NH), 29.42 (C-7,
*CH2CH2CH2CH2NH), 29.72 (C-9, CH2CH2*CH2CH2NH), 48.14 (C-10, CH2CH2CH2*CH2NH), 57.10
(C-3, CHNH), 61.78 (C-5, CH2NH ), 65.5 (C-14, OCH2), 128.18 (C-17, aromatic), 128.81 (C-16,

10
aromatic), 137.77 (C-15, aromatic), 156.55 (C-12, *CO2CH2Ph), 173.88 (C-6,*COCH2NH), 175.45 (C-
2,*COCHNH). (m/z): 320.2.

(3S)-tert-butyl-(4-(3,5-dioxopiperazin-2-yl)butyl)carbamate
(13aa). Yield: 74% yellow solid. mp: 113-115 °C. IR υmax
(ATR) cm-1: 1H NMR (400 MHz, CDCl3) δ: 1.44 (H9, s, H-15,
CO2CH3), 1.53 (4H, m, CH2*CH2*CH2CH2NH, H-8, H-9),
1.69 (1H, m, H-7B, *CH2CH2CH2CH2NH), 2.01 (2H, m, H-4
y H-7A, NH, *CH2CH2CH2CH2NH), 3.14 (2H, d, J=5.4 Hz,
H-10, CH2CH2CH2*CH2NH), 3.42 (1H, dd, J=4.9 y 8.8 Hz, H-3,*CHNH), 3.58 (1H, d, J=18 Hz, H-5B,
*CH2NH ), 3.74 (1H, d, J=18 Hz, H-5A, *CH2NH ), 4.86 (1H, br, H-11, COONH ), 8.66 (1H, br, H-1,
CONHCO), 13
C-NMR (100 MHz, CDCl3) δ: 22.84 (C-8, CH2*CH2CH2CH2NH), 28.41 (C-15,
OC(*CH3)3), 29.33 (C-7, *CH2CH2CH2CH2NH), 29.77 (C-9, CH2CH2*CH2CH2NH), 40.13(C-10,
CH2CH2CH2*CH2NH), 47.92 (C-5, CH2NH), 57.5 (C-3, CHNH ), 79.19 (C-14, O*C(CH3)3), 156.08 (C-
12, *CO2C(CH3)3), 172.10 (C-6,*COCH2NH), 173.74 (C-2,*COCHNH). (m/z): 286.2.

(3S)-3-((1H-indol-2-yl)methyl)piperazine-2,6-dione (16a). Yield: 83%

white solid. mp: 183-185 °C. IR υmax (ATR) cm-1: 3411, 3283, 2979,
2934, 2721, 1711, 1691. 1H NMR (500 MHz, DMSO-d6) δ: 2.73 (1H,
dt, J=9 y 14.2 Hz, H-4, NH), 2.93 (1H, dd, J= 8.6 Hz, J= 14.6 Hz, H-
7B, *CH2-indol), 3.24 (1H, dd, J= 2.75 y 14.6 Hz, H-7A, *CH2-indol),
3.34 (1H, overlapped with H2O, H-5B, *CH2NH), 3.43 (1H, dd, J= 4.4 y
17.3 Hz H-5A, *CH2NH), 3.59 (1H, m o td, J= 4.4 y 16.5 Hz H-3,
*CHNH ), 6.94 (1H, t, J= 7.3 Hz, H-14, aromatic), 7.03 (1H, t, J= 7.1
Hz, H-13, aromatic), 7.14 (1H, s, H-9, CH=*CHNH-indol), 7.31 (1H, d, J= 8 Hz, H-15, aromatic), 7.52
(1H, d, J=7.7 Hz, H
-12, aromatic), 10.82 (2H, br, H-1, H10, CONHCO, NH-indol). 13C-NMR (100 MHz, DMSO-d6) δ: 25.76
(C-7, *CH2-indol), 48.56 (C-5, *CH2NH), 58.39 (C-3, *CHCONH), 110.88 (C-8, *CH=CHNH-indol),
111.84 (C-15, aromatic), 118.81 (C-14, aromatic), 119.0 (C-12, aromatic), 121.36 (C-13, aromatic),
124.46 (C-9, CH=*CHNH-indol), 128.01 (C-16, aromatic), 136.60 (C-11, aromatic), 173.91 (C-

11
6,*COCH2NH), 175.17 (C-2,*COCHNH). HR-ESI/TOF-MS [M+H]+ (m/z) cal for C13H14N3O2 :
244.108053, found: 244.108455.

(3R)-3-((1H-indol-2-yl)methyl)piperazine-2,6-dione (16b). Yield:

68%. mp: 182-184 °C. IR υmax (ATR) cm-1: 3411, 3282, 2977, 2934,
2711, 1711, 1691. 1H NMR (500 MHz, DMSO-d6) δ: 2.73 (1H, dt, J=9 y
14.2 Hz, H-4, NH), 2.93 (1H, dd, J= 8.6 Hz, J= 14.6 Hz, H-7B, *CH2-
indol), 3.24 (1H, dd, J= 2.75 y 14.6 Hz, H-7A, *CH2-indol), 3.34 (1H,
overlapped with H2O, H-5B, *CH2NH), 3.43 (1H, dd, J= 4.4 y 17.3 Hz
H-5A, *CH2NH), 3.59 (1H, m o td, J= 4.4 y 16.5 Hz H-3, *CHNH ),
6.94 (1H, t, J= 7.3 Hz, H-14, aromatic), 7.03 (1H, t, J= 7.1 Hz, H-13, aromatic), 7.14 (1H, s, H-9,
CH=*CHNH-indol), 7.31 (1H, d, J= 8 Hz, H-15, aromatic), 7.52 (1H, d, J=7.7 Hz, H
-12, aromatic), 10.82 (2H, br, H-1, H10, CONHCO, NH-indol). 13C-NMR (100 MHz, DMSO-d6) δ: 25.76
(C-7, *CH2-indol), 48.56 (C-5, *CH2NH), 58.39 (C-3, *CHCONH), 110.88 (C-8, *CH=CHNH-indol),
111.82 (C-15, aromatic), 118.79 (C-14, aromatic), 118.98 (C-12, aromatic), 121.34 (C-13, aromatic),
124.44 (C-9, CH=*CHNH-indol), 128.01 (C-16, aromatic), 136.60 (C-11, aromatic), 173.88 (C-
6,*COCH2NH), 175.16 (C-2,*COCHNH). HR-ESI/TOF-MS [M+H]+ (m/z) cal for C13H14N3O2:
244.108053, found: 244.108262.

Results and Discussion

Conclusions

Acknowledgments

The author F.P. G-G thanks to CONACYT for scholarship granted during the development of
this work.

References

12