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tanediamide-like compound 12. In conclusion, with the same ami-
References and notes
no acid sequence, the linear tetrapeptide is the weakest, cyclo tet-
rapeptide is somewhat stronger and the heptanediamide-like
compound is the strongest inhibitor, respectively. When compared
to the known cyclotetrapeptide Apicidin (IC50 = 4 nM)32 and Trap-
oxin B (IC50 = 0.11 nM for HDAC1),33 the activities of our com-
pounds have decreased considerably. Optimal activity was
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observed with compound 10 (IC50 = 4.7
lM for HDAC1 and
1.3 M for HDAC3). Surprisingly, compound 15 shows a 19-fold
l
selectivity between HDAC1 and HDAC3, with a clear preference
for HDAC3.
The effects of 1, 10, 11, 12, 13, 14, 15, and 16 (at 20
concentration) on the acetylation levels of endogenous histone
H3 in human lymphoblastoid cell line (GM15850, Coriell
lM
a
Institute) were determined by western blot analysis, after
48 h incubations. The result of this experiment is shown in
Fig. 5. For the H4 RGKG mimics, 10 showed relatively stronger
effects on acetylated H3, 13 showed no effect and 1 showed
moderate effects on acetylation. These results are consistent
with the IC50 values for recombinant HDACs (Table 2). As for
the trapoxin mimics, the same trend was observed; that is,
the lower the IC50 value for each compound (Table 2), the
stronger the effect on histone H3 acetylation in cells. As for
the amino acid inhibitors, compound 15 showed a relatively
strong effect on cellular histone H3 acetylation, while com-
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tively charged Arg group in compound 16 may hinder the
compound to penetrate cells, while the relative neutral com-
pound 15 may allow cell penetration. This difference in charge
may explain why compounds 15 and 16 have remarkable dif-
ferences in their ability to cause histone H3 acetylation in cells
but nearly equal IC50 values for inhibition of recombinant
HDAC3, and compound 15 has a higher IC50 for HDAC1 than
compound 16.
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In conclusion, we have synthesized two different types of benz-
amide compounds. Most show moderate inhibitory activity on
HDAC1 and HDAC3. Compound 15 shows 19-fold selectivity be-
tween HDAC3 and HDAC1. Further studies will be carried out on
compound 15.
Acknowledgments
This work was supported in part by National Institutes of
Health Grant R21-NS055781, and by Repligen Corporation, Wal-
tham, MA.
33. Furumai, R.; Komatsu, Y.; Nishino, N.; Khochbin, S.; Yoshida, M.; Horinouchi, S.
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