6840
X. Teng et al. / Bioorg. Med. Chem. Lett. 17 (2007) 6836–6840
6. Lo, E. H.; Dalkara, T.; Moskowitz, M. A. Nat. Rev.
Neurosci. 2003, 4, 399.
7. McCully, J. D.; Wakiyama, H.; Hsieh, Y. J.; Jones, M.;
Levitsky, S. Am. J. Physiol. Heart Circ. Physiol. 2004, 286,
H1923.
8. Yuan, J.; Lipinski, M.; Degterev, A. Neuron 2003, 40, 401.
9. Teng, X.; Degterev, A.; Jagtap, P.; Xing, X.; Choi, S.;
Denu, R.; Yuan, J.; Cuny, G. D. Bioorg. Med. Chem. Lett.
2005, 15, 5039.
pharmacology of these compounds in animal models of
disease where necroptosis is likely to play a substantial
role (i.e. cerebral ischemia, traumatic brain injury, and
liver injury). Additionally, these compounds are being
used to further interrogate the mechanism(s) of necrop-
totic cell death.
Acknowledgments
10. Jagtap, P. G.; Degterev, A.; Choi, S.; Keys, H.; Yuan, J.;
Cuny, G. D. J. Med. Chem. 2007, 50, 1886.
11. Wang, K.; Li, J.; Degterev, A.; Hsu, E.; Yuan, J.; Yuan,
C. Bioorg. Med. Chem. Lett. 2007, 17, 1455.
X.T. and G.D.C. thank the Harvard Center for Neuro-
degeneration and Repair (HCNR) for financial support.
A.D. and J.Y. thank the National Institute on Aging,
National Institute of General Medical Sciences and
American Health Assistance Foundation for financial
support. X.T., G.D.C., and J.Y. thank the National
Institute of Neurological Disorders and Stroke
(NINDS) for financial support. J.A.P. Jr. thanks the
National Institutes of Health and Bristol-Myers Squibb
for financial support. A.D. is a recipient of NIH Men-
tored Scientist Development Award from the National
Institute on Aging (NIA). The SV40-transformed adult
mouse lung fibroblasts were a generous gift of Dr. Philip
Tsichlis (Tufts University).
12. Several other biological activities of 4-alkyl-[1,2,3]thiadi-
azole-5-benzylamides have been reported, including inhi-
bition of I-kappa B kinase complex (IKK), see: Pitts, W.
J.; Kempson, J.; Guo, J.; Das, J.; Langevine, C. M.;
Spergel, S. H.; Watterson, S. H. WO 2006122137, 2006
and as agents to control plant diseases; See: Umetani, K;
Shimaoka, T.; Yamaguchi, M.; Oda, M.; Kyomura, N.;
Takemoto, T.; Kikutake, K. WO 2006098128, 2006.
13. Oikawa, Y.; Sugano, K.; Yonemitsu, O. J. Org. Chem.
1978, 43, 2087.
14. Thomas, E. W.; Nishizawa, E. E.; Zimmermann, D. C.;
Williams, D. J. J. Med. Chem. 1985, 28, 442.
15. Polniaszek, R. P.; Belmont, S. E.; Alvarez, R. J. Org.
Chem. 1990, 55, 215.
16. Itsuno, S.; Hachisuka, C.; Ito, K. J. Chem. Soc., Perkin
Trans. 1 1991, 1767.
17. Lebel, H.; Leogane, O. Org. Lett. 2005, 7, 4107.
18. Burk, M. J.; Wang, Y. M.; Lee, J. R. J. Am. Chem. Soc.
1996, 118, 5142.
References and notes
1. Wyllie, A. H.; Kerr, J. F. R.; Currie, A. R. Int. Rev. Cytol.
1980, 68, 251.
19. For EC50 value determinations, FADD-deficient variant
of human Jurkat T cells (5 · 105 cells/mL, 100 lL per well
in a 96-well plate) was treated with 10 ng/mL of human
TNF-a in the presence of increasing concentration of test
compounds for 24 h at 37 ꢁC in a humidified incubator
with 5% CO2 followed by ATP-based viability assessment.
Stock solutions (30 mM) in DMSO were prepared and
then diluted with DMSO to give testing solutions, which
were added to each test well. The final DMSO concentra-
tion was 0.5%. Eleven compound test concentrations
(0.030–100 lM) were used. Each concentration was done
in duplicate. Cell viability assessments were performed
using a commercial luminescent ATP-based assay kit
(CellTiter-Glo) according to the manufacturer’s instruc-
tions. Cell lysis/ATP detection reagent (40 lL) was added
to each well. Plates were incubated on a rocking platform
for 10 min at room temperature and luminescence was
measured using a Wallac Victor 3 plate-reader (Perkin
Elmer). Cell viability was expressed as a ratio of the signal
in the well treated with TNF-a and compound to the
signal in the well treated with compound alone. This was
done to account for nonspecific toxicity, which in most
cases was <10%. EC50 values were calculated using
nonlinear regression analysis of sigmoid dose–response
(variable slope) curves from plots of log[I] verses viability
values.
2. (a) Cryns, V.; Yuan, J. Genes Dev. 1998, 12, 1551; (b)
Yuan, J.; Yankner, B. A. Nature 2000, 407, 802; (c)
Talanian, R. V.; Brady, K. D.; Cryns, V. L. J. Med. Chem.
2000, 43, 3351; (d) Moore, J. D.; Rothwell, N. J.; Gibson,
R. M. Br. J. Pharmacol. 2002, 135, 1069, and references
therein; (e) Boyce, M.; Degterev, A.; Yuan, J. Cell Death
Differ. 2004, 11, 29.
3. For literature related to caspase-independent cell death,
see (a) Kitanaka, C.; Kuchino, Y. Cell Death Differ. 1999,
6, 508; (b) Fiers, W.; Beyaert, R.; Declercq, W.; Vanden-
abeele, P. Oncogene 1999, 18, 7719; (c) Borner, C.;
Monney, L. Cell. Death Differ. 1999, 6, 497; (d) Edinger,
A. L.; Thompson, C. B. Curr. Opin. Cell Biol. 2004, 16,
663; (e) Yu, L.; Alva, A.; Su, H.; Dutt, P.; Freundt, E.;
Welsh, S.; Baehrecke, E. H.; Lenardo, M. J. Science 2004,
304, 1500; (f) Chipuk, J. E.; Green, D. R. Nat. Rev. Mol.
Cell Biol. 2005, 6, 268; (g) Bro¨ker, L. E.; Kruyt, F. A. E.;
Giaccone, G. Clin. Cancer Res. 2005, 11, 3155; (h) Fink, S.
L.; Cookson, B. T. Infect. Immun. 2005, 73, 1907; (i)
Kroemer, G.; Martin, S. J. Nat. Med. 2005, 11, 725; (j)
Vandenabeele, P.; Vanden Berghe, T.; Festjens, N. Sci.
STKE 2006, 358, pe44; (k) Martinet, W.; Schrijvers, D.
M.; Herman, A. G.; De Meyer, G. R. Autophagy 2006, 2,
312.
4. Degterev, A.; Huang, Z.; Boyce, M.; Li, Y.; Jagtap, P.;
Mizushima, N.; Cuny, G. D.; Mitchison, T.; Moskowitz,
M.; Yuan, J. Nat. Chem. Biol. 2005, 1, 112.
5. Martin, L. J.; Al-Abdulla, N. A.; Brambrink, A. M.;
Kirsch, J. R.; Sieber, F. E.; Portera-Cailliau, C. Brain Res.
Bull. 1998, 46, 281.
20. Compound 52 was prepared in low yield (10%) by
allowing 16 to react with 2,6-difluoroaniline in THF and
pyridine at room temperature. The reaction with 2-chloro-
6-fluoroaniline was unsuccessful presumably due to
increased steric hindrance.