S. Nasim et al. / Bioorg. Med. Chem. Lett. 21 (2011) 4879–4883
4883
8. Stanton, M. G.; Gagne, M. R. J. Org. Chem. 1997, 62, 8240.
9. Andrew, D. R.; Gaeta, F. C. U.S. Patent 4885,293, 1989.
NMR (75 MHz, CD3OD) d ppm 165.7, 153.2, 137.4, 135.1, 129.0, 128.7, 46.9,
46.4, 44.1, 41.7; Anal. (C12H15Cl2N3O2S) C, H, N. 2-(20-Bromoethyl)-4-[400-
methylaminobenzyl)-1,2,4-thiadiazol-idine-3,5-dione hydrochloride (10f), white
solid, mp = 91–93 °C, yield = 75%; 1H NMR (300 MHz, CD3OD) d ppm 7.44 (s,
10. Spectral and analytical data for compounds 3a, 3b and 3e were in agreement
with previously reported data.3b,c,6 2-Propyl-4-benzyl-1,2,4-thiadiazolidine-3,5-
dione (3c), clear oil, yield = 72%; 1H NMR (300 MHz, CDCl3) d ppm 7.42–7.23 (m,
5H), 4.79 (s, 2H), 3.56 (t, J = 6.5 Hz, 2H), 1.61 (q, J = 6.5 Hz, 2H), 0.92 (t,
J = 6.5 Hz, 3H); 13C NMR (75 MHz, CDCl3) d ppm 166.0, 153.0, 135.3, 128.9,
128.8, 128.3, 46.8, 46.2, 22.4, 11.4; EI-MS m/z = 250; Anal. (C12H14N2O2S) C, H,
4H), 4.85 (s, 2H), 4.10 (s, 2H), 4.04 (t, J = 6.3 Hz, 2H), 3.63 (t, J = 6.3 Hz, 2H); 13
C
NMR (75 MHz, CD3OD) d ppm 167.3, 154.6, 137.9, 134.2, 130.3, 129.8, 47.5,
46.3, 44.0, 30.5; Anal. (C12H15BrClN3O2S) C, H, N. 2-(20-Chloroethyl)-4-(300-
methylaminobenzyl)-1,2,4-thiadiazolidine-3,5-dione hydro-chloride (10g), white
solid, mp 97–99 °C, yield = 53%; 1H NMR (300 MHz, CD3OD) d ppm 7.36 (t,
J = 7.6 Hz, 1H), 7.31 (m, 3H), 4.83 (s, 2H), 4.13 (s, 2H), 4.05 (t, J = 5.7 Hz, 2H),
3.81 (t, J = 5.7 Hz, 2H); 13C NMR (75 MHz, CD3OD) d ppm 165.7, 153.2, 135.1,
129.0, 128.9, 128.7, 128.3, 128.5, 46.5, 46.3, 43.9, 42.1; Anal.
(C12H15Cl2N3O2SꢀH2O) C, H, N. 4-{40-[200-(2000-Chloroethyl)-1,2,4-thiadiazolidine-
N. 2-Allyl-4-benzyl-1,2,4-thiadiazolidine-3,5-dione (3d), clear oil, yield = 60%; 1
H
NMR (300 MHz, CDCl3) d ppm 7.46–7.26 (m, 5H), 5.81 (m, 1H), 5.35 (m, 2H),
4.83 (s, 2H), 4.23 (m, 2H); 13C NMR (75 MHz, CDCl3) d ppm 165.9, 152.6, 135.2,
131.0, 128.9, 128.7, 128.3, 120.7, 47.4, 46.1; EI-MS m/z = 248. Anal.
(C12H12N2O2S) C, H, N. 2-(20-Bromoethyl)-4-benzyl-1,2,4-thiadiazolidine-3,5-
dione (3f), white solid, mp = 76–78 °C, yield = 82%; 1H NMR (300 MHz, CDCl3)
d ppm 7.45–7.30 (m, 5H), 4.83 (s, 2H), 3.99 (t, J = 6.3 Hz, 2H), 3.52 (t, J = 6.3 Hz,
2H); 13C NMR (75 MHz, CDCl3) d ppm 165.7, 153.2, 135.1, 129.0, 128.9, 128.5,
46.9, 46.4, 29.1. Anal. (C11H11BrN2O2S) C, H, N. 2-Ethyl-4-(40-
methylaminobenzyl)-1,2,4-thiadiazolidine-3,5-dione hydro-chloride (10b), white
solid, mp = 151–153 °C, yield = 73%; 1H NMR (300 MHz, CD3OD) d ppm 7.44 (s,
3,5-dionyl]} sodium benzoate (15), white solid, mp = 170–172 °C, yield = 90%; 1
H
NMR (300 MHz, CD3OD) d ppm 7.97 (d, J = 8.4 Hz, 2H), 7.43 (d, J = 8.4 Hz, 2H),
4.74 (s, 2H), 3.96 (t, J = 5.4 Hz, 2H), 3.73 (t, J = 5.4 Hz, 2H); 13C NMR (75 MHz,
CD3OD) d ppm 168.9, 167.0, 154.1, 140.9, 131.2, 130.8, 128.8, 47.4, 46.2, 42.6.
Anal (C12H10ClN2O4SNa) C, H, N.
11. Assessment of LD50 for TDZD analogs from MV4-11 cell assays. MV4-11 cells
were purchased from the ATCC (American type culture collection). Cells were
cultured at 300,000 cells/mL and exposed to at least 4 different concentrations
of the various analogs for 24 h (or for the indicated time period in the time-
course experiments). Iscove’s Modified Dulbecco’s Medium (IMDM)
supplemented with 10% fetal bovine serum (FBS) was used. Cell viability was
determined by staining with Annexin V-Fluorescein isothiocyanate (FITC) and
7-aminoactinomycin D (7-AAD). Analyses were performed in the BD-LSRII
15 min later. The percent of viable cells was determined relative to the
untreated control by gaiting on Annexin V negative/7-AAD negative cells. The
calculation of LD50 values was performed using Calcusyn software (Biosoft).
4H), 4.83 (s, 2H), 4.10 (s, 2H), 3.69 (q, J = 6.3 Hz, 2H), 1.23 (t, J = 6.3 Hz, 3H); 13
C
NMR (75 MHz, CD3OD) d ppm 166.4, 152.9, 136.5, 134.3, 129.9, 128.3, 45.5,
42.5, 39.8, 14.3. Anal. (C12H16ClN3O2Sꢀ0.5H2O) C, H, N. 2-Allyl-4-(40-
methylaminobenzyl)-1,2,4-thiadiazolidine-3,5-dione hydrochloride (10d), white
solid, mp = 105–107 °C, yield = 75%; 1H NMR (300 MHz, CD3OD) d ppm 7.44 (s,
4H), 5.87 (m, 1H), 5.32 (m, 2H), 4.85 (s, 2H), 4.24 (m, 2H), 4.10 (s, 2H); 13C NMR
(75 MHz, CD3OD) d ppm 167.7, 154.6, 138.4, 134.5, 132.9, 130.6, 130.3, 120.6,
48.4,46.6, 44.3; Anal (C13H16ClN3O2S) C, H, N. 2-(20-Chloroethyl)-4-(400-
methylaminobenzyl)-1,2,4-thiadiazolidine-3,5-dione hydrochloride (10e), white
solid, mp = 103–105 °C, yield = 64%; 1H NMR (300 MHz, CD3OD) d ppm 7.42 (s,
4H), 4.82 (s, 2H), 4.15 (s, 2H), 3.90 (t, J = 6.5 Hz, 2H), 3.78 (t, J = 6.5 Hz, 2H); 13
C