Page 5 of 6
ACS Medicinal Chemistry Letters
TEFb facilitates oncogenic and physiologic MLLꢀdependent transcription.
Cancer Cell 2010, 17, 198ꢀ212.
(6) Mohan, M.; Herz, H. M.; Takahashi, Y. H.; Lin, C.; Lai, K. C.; Zhang,
Y.; Washburn, M. P.; Florens, L.; Shilatifard, A. Linking H3K79
trimethylation to Wnt signaling through a novel Dot1ꢀcontaining complex
(DotCom). Genes Dev 2010, 24, 574ꢀ589.
with a Ki value of 56 nM and is only 2ꢀ3 times less potent to
AF9, but 28 binds to AFꢀ9 13ꢀfold more potently than to ENL,
suggesting that Nꢀterminal modifications could alter the selecꢀ
tivity in binding to the two fusion proteins.
1
2
3
4
5
6
7
8
9
In summary, based on the 7 mer DOT1L peptide, a series of
peptidomimetics was designed and synthesized to improve the
binding affinity to AF9 and decrease the peptidic characterisꢀ
tics. By optimizing the middle three residues we identified
peptide 21 which has a significantly improved binding affinity
compared to the original peptide 2. Based on 21, we have perꢀ
formed preliminary modifications to both the C- and Nꢀtermini
of 21. For Cꢀterminal dipeptide motif we found that the amide
bond in the dipeptide can be replaced with an imidazole ring
and hydrophobic groups can be introduced to the imidazole
ring to mimic the hydrophobic interaction of the DOT1L leuꢀ
cine residue L871. For the Nꢀterminal dipeptide motif, reꢀ
placement of the leucine residue L865 with a suitable hydroꢀ
phobic group can improve the binding affinity. Detailed modiꢀ
fications to the Cꢀ and Nꢀterminal residues and the mechanistic
studies for the designed compounds are proceeding and the
results will be reported subsequently.
(7) Tan, J.; Jones, M.; Koseki, H.; Nakayama, M.; Muntean, A. G.;
Maillard, I.; Hess, J. L. CBX8, a polycomb group protein, is essential for
MLLꢀAF9ꢀinduced leukemogenesis. Cancer Cell 2011, 20, 563ꢀ575.
(8) Steger, D. J.; Lefterova, M. I.; Ying, L.; Stonestrom, A. J.; Schupp,
M.; Zhuo, D.; Vakoc, A. L.; Kim, J. E.; Chen, J.; Lazar, M. A.; Blobel, G.
A.; Vakoc, C. R. DOT1L/KMT4 recruitment and H3K79 methylation are
ubiquitously coupled with gene transcription in mammalian cells. Mol
Cell Biol 2008, 28, 2825ꢀ2839.
(9) Luo, Z.; Lin, C.; Guest, E.; Garrett, A. S.; Mohaghegh, N.; Swanson,
S.; Marshall, S.; Florens, L.; Washburn, M. P.; Shilatifard, A. The super
elongation complex family of RNA polymerase II elongation factors: gene
target specificity and transcriptional output. Mol Cell Biol 2012, 32, 2608ꢀ
2617.
(10) Shen, C.; Jo, S. Y.; Liao, C.; Hess, J. L.; NikolovskaꢀColeska, Z.
Targeting recruitment of disruptor of telomeric silencing 1ꢀlike (DOT1L):
characterizing the interactions between DOT1L and mixed lineage
leukemia (MLL) fusion proteins. J Biol Chem 2013, 288, 30585ꢀ30596.
(11) Leach, B. I.; Kuntimaddi, A.; Schmidt, C. R.; Cierpicki, T.; Johnson,
S. A.; Bushweller, J. H. Leukemia fusion target AF9 is an intrinsically
disordered transcriptional regulator that recruits multiple partners via
coupled folding and binding. Structure 2013, 21, 176ꢀ183.
(12) Krivtsov, A. V.; Feng, Z.; Lemieux, M. E.; Faber, J.; Vempati, S.;
Sinha, A. U.; Xia, X.; Jesneck, J.; Bracken, A. P.; Silverman, L. B.;
Kutok, J. L.; Kung, A. L.; Armstrong, S. A. H3K79 methylation profiles
define murine and human MLLꢀAF4 leukemias. Cancer cell 2008, 14,
355ꢀ368.
(13) Bernt, K. M.; Zhu, N.; Sinha, A. U.; Vempati, S.; Faber, J.; Krivtsov,
A. V.; Feng, Z.; Punt, N.; Daigle, A.; Bullinger, L.; Pollock, R. M.;
Richon, V. M.; Kung, A. L.; Armstrong, S. A. MLLꢀrearranged Leukemia
is Dependent on Aberrant H3K79 Methylation by DOT1L. Cancer Cell
2011, 20, 66ꢀ78.
(14) Nguyen, A. T.; Taranova, O.; He, J.; Zhang, Y. DOT1L, the H3K79
methyltransferase, is required for MLLꢀAF9–mediated leukemogenesis.
Blood 2011, 117, 6912ꢀ6922.
(15) Daigle, S. R.; Olhava, E. J.; Therkelsen, C. A.; Majer, C. R.;
Sneeringer, C. J.; Song, J.; Johnston, L. D.; Scott, M. P.; Smith, J. J.;
Xiao, Y.; Jin, L.; Kuntz, K. W.; Chesworth, R.; Moyer, M. P.; Bernt, K.
M.; Tseng, J.ꢀC.; Kung, A. L.; Armstrong, S. A.; Copeland, R. A.; Richon,
V. M.; Pollock, R. M. Selective Killing of Mixed Lineage Leukemia Cells
by a Potent SmallꢀMolecule DOT1L Inhibitor. Cancer cell 2011, 20, 53ꢀ
65.
10
11
12
13
14
15
16
17
18
19
20
21
22
23
24
25
26
27
28
29
30
31
32
33
34
35
36
37
38
39
40
41
42
43
44
45
46
47
48
49
50
51
52
53
54
55
56
57
58
59
60
ASSOCIATED CONTENT
Supporting Information
The experimental details for the synthesis of the designed comꢀ
pounds, characterization data, fluorescence polarization (FP)
binding assay, bioꢀlayer interferometry (BLI) binding studies,
pullꢀdown and coꢀIP experiments. The Supporting Information is
available free of charge on the ACS Publications website.
AUTHOR INFORMATION
Corresponding Authors
gmail.com (H.S.)
Author Contributions
‡These authors contributed equally.
Funding Sources
(16) Nguyen, A. T.; Zhang, Y. The diverse functions of Dot1 and H3K79
methylation. Genes & Development 2011, 25, 1345ꢀ1358.
This work was supported by the UM Center for Discovery of New
Medicine (CDNM) to Z.NꢀC. The National Science Foundation
Graduate Research Fellowship Program and Rackham Merit Felꢀ
lowship to S.M.G. The Program for Jiangsu Province Innovative
Research Team and the Program for Specially Appointed Profesꢀ
sor of Jiangsu Province to H.S.
(17) Jo, S. Y.; Granowicz, E. M.; Maillard, I.; Thomas, D.; Hess, J. L.
Requirement for Dot1l in murine postnatal hematopoiesis and
leukemogenesis by MLL translocation. Blood 2011, 117, 4759ꢀ4768.
(18) Kuntimaddi, A.; Achille, N. J.; Thorpe, J.; Lokken, A. A.; Singh, R.;
Hemenway, C. S.; Adli, M.; ZeleznikꢀLe, N. J.; Bushweller, J. H. Degree
of recruitment of DOT1L to MLLꢀAF9 defines level of H3K79 Diꢀ and
triꢀmethylation on target genes and transformation potential. Cell Rep
2015, 11, 808ꢀ820.
(19) Watson, V. G.; Drake, K. M.; Peng, Y.; Napper, A. D. Development
of a highꢀthroughput screeningꢀcompatible assay for the discovery of
inhibitors of the AF4ꢀAF9 interaction using AlphaScreen technology.
Assay Drug Dev Technol 2013, 11, 253ꢀ268.
Notes
The authors declare no competing financial interest.
REFERENCES
(1) Muntean, A. G.; Hess, J. L. The pathogenesis of mixedꢀlineage
leukemia. Annu Rev Pathol 2012, 7, 283ꢀ301.
(2) Yokoyama, A. Transcriptional activation by MLL fusion proteins in
leukemogenesis. Exp Hematol 2017, 46, 21ꢀ30.
(3) Krivtsov, A. V.; Armstrong, S. A. MLL translocations, histone
modifications and leukaemia stemꢀcell development. Nature Reviews
Cancer 2007, 7, 823ꢀ833.
(4) Lin, C.; Smith, E. R.; Takahashi, H.; Lai, K. C.; MartinꢀBrown, S.;
Florens, L.; Washburn, M. P.; Conaway, J. W.; Conaway, R. C.;
Shilatifard, A. AFF4, a component of the ELL/PꢀTEFb elongation
complex and a shared subunit of MLL chimeras, can link transcription
elongation to leukemia. Mol Cell 2010, 37, 429ꢀ437.
(5) Yokoyama, A.; Lin, M.; Naresh, A.; Kitabayashi, I.; Cleary, M. L. A
higherꢀorder complex containing AF4 and ENL family proteins with Pꢀ
5
ACS Paragon Plus Environment