Design of the First-in-Class, Highly Potent Irreversible Inhibitor Targeting the Menin-MLL Protein–Protein Interaction

Article abstract of DOI:10.1002/anie.201711828

The structure-based design of M-525 as the first-in-class, highly potent, irreversible small-molecule inhibitor of the menin-MLL interaction is presented. M-525 targets cellular menin protein at sub-nanomolar concentrations and achieves low nanomolar potencies in cell growth inhibition and in the suppression of MLL-regulated gene expression in MLL leukemia cells. M-525 demonstrates high cellular specificity over non-MLL leukemia cells and is more than 30 times more potent than its corresponding reversible inhibitors. Mass spectrometric analysis and co-crystal structure of M-525 in complex with menin firmly establish its mode of action. A single administration of M-525 effectively suppresses MLL-regulated gene expression in tumor tissue. An efficient procedure was developed to synthesize M-525. This study demonstrates that irreversible inhibition of menin may be a promising therapeutic strategy for MLL leukemia.

Full text of DOI:10.1002/anie.201711828

A Journal of the Gesellschaft Deutscher Chemiker
Angewandte
Chemie
International Edition
www.angewandte.org
Accepted Article
Title: Design of the First-in-Class, Highly Potent Irreversible Inhibitor
Targeting the Menin-MLL Protein-Protein Interaction
Authors: Shaomeng Wang, Shilin Xu, Angelo Aguilar, Tianfeng Xu,
Ke Zheng, Liyue Huang, Jeanne Stuckey, Krishnapriya
Chinnaswamy, Denzil Bernard, Ester Fernández-Salas, Liu
Liu, Mi Wang, Donna McEachern, Sally Przybranowski, and
Caroline Foster
This manuscript has been accepted after peer review and appears as an
Accepted Article online prior to editing, proofing, and formal publication
of the final Version of Record (VoR). This work is currently citable by
using the Digital Object Identifier (DOI) given below. The VoR will be
published online in Early View as soon as possible and may be different
to this Accepted Article as a result of editing. Readers should obtain
the VoR from the journal website shown below when it is published
to ensure accuracy of information. The authors are responsible for the
content of this Accepted Article.
To be cited as: Angew. Chem. Int. Ed. 10.1002/anie.201711828
Angew. Chem. 10.1002/ange.201711828
Link to VoR: http://dx.doi.org/10.1002/anie.201711828
http://dx.doi.org/10.1002/ange.201711828

10.1002/anie.201711828
Angewandte Chemie International Edition
COMMUNICATION
Design of A First-in-Class, Highly Potent Irreversible Inhibitor
Targeting the Menin-MLL Protein-Protein Interaction
Shilin Xu+, Angelo Aguilar+, Tianfeng Xu+, Ke Zheng+, Liyue Huang+, Jeanne Stuckey, Krishnapriya
Chinnaswamy, Denzil Bernard, Ester Fernández-Salas, Liu Liu, Mi Wang, Donna McEachern, Sally
Przybranowski, Caroline Foster, Shaomeng Wang*
Abstract: We report the structure-based design of M-525 (10), a first-
in-class, highly potent, irreversible small-molecule inhibitor of the
menin-MLL protein-protein interaction. M-525 targets cellular menin
protein at sub-nanomolar concentrations and achieves low nanomolar
potencies in cell growth inhibition and in suppression of MLL-
regulated gene expression in MLL leukemia cells. It demonstrates
high cellular specificity over non-MLL leukemia cells and is >30-times
more potent than the corresponding reversible inhibitors. Mass
spectroscopic analysis and the co-crystal structure of M-525 in a
complex with menin firmly establish its mode of action. A single
administration of M-525 effectively suppresses MLL-regulated gene
expression in tumor tissue. An efficient synthetic procedure was
developed to obtain enantiomerically pure M-525 and its analogues.
Our study demonstrates that irreversible inhibition of menin may
represent a promising therapeutic strategy for MLL leukemia.
characterization of the first-in-class, highly potent and irreversible
menin inhibitors.
For the design of irreversible menin inhibitors, we sought first
to obtain a highly potent reversible menin inhibitor. We used MIV-
6 (1), a previously reported, moderately potent, reversible
inhibitor[4c]
, as the starting point for our structure-based
optimization efforts, which are summarized in Figure 1.
MLL leukemia, which is characterized by chromosomal
translocations at 11q23 and expression of MLL fusion proteins,
has a very poor prognosis and is resistant to current therapies[1].
MLL fusion proteins interact directly with menin, a protein
encoded by the MEN1 gene, to regulate the expression of MEIS1
and HOX genes, which drive leukemogenesis in MLL leukemia[2].
Consequently, targeting the menin-MLL protein-protein
interaction using small-molecule inhibitors (here called menin
inhibitors) is a promising therapeutic strategy for MLL leukemia[3].
Several classes of peptidomimetic compounds and non-
peptide small-molecule menin inhibitors have been reported in
recent years[4]. To date, all published menin inhibitors are
reversible in nature. Preclinical data show that extended drug
exposure is required for menin inhibitors to achieve anti-leukemia
activity in vitro[4d]. We hypothesized that since irreversible
inhibitors in general can achieve target inhibition superior to that
of reversible inhibitors[5], irreversible menin inhibitors may have
much greater anti-leukemia activity and efficacy than reversible
menin inhibitors. In this paper, we report the design and
Figure 1. Structure-based design of irreversible menin inhibitors and control
compounds. Compounds 5~11 are enantiopure.
The primary amino group in MIV-6 was shown to be a major
metabolic softspot and in addition, the synthetic yield for the “core”
structure bearing this primary amino group was very low[4c]
.
Consequently, we replaced this free amino group in MIV-6 with a
metabolically stable nitrile group, producing compound 2, which
binds to menin with an IC50 value of 3.1 µM (Table 1). Although 2
is 17-times less potent than MIV-6 in binding to menin, its
synthetic yield is high, and it represents a useful starting point for
further optimization.
MIV-6 and compound 2 have a flexible linker connecting their
respective “core” structure to the same “tail” group. Because
restriction of rotatable bonds is an effective means of reducing
conformational space and potential off-target activity of small-
molecule drugs, we restricted the linker conformation using 3-
methylazetidine. This led to compound 3 that binds to menin with
IC50 = 3.0 µM and is as potent as compound 2.
[*]
Dr. S. Xu[+], Dr. A. Aguilar[+], Dr. T. Xu[+], Dr. K. Zheng[+], Dr. L.
Huang[+], Dr. D. Bernard, Dr. L. Liu, Dr. M. Wang, D. McEachern, S.
Przybranowski, C. Foster, Prof. Dr. S. Wang
Comprehensive Cancer and Departments of Internal Medicine,
Pharmacology and Medicinal Chemistry, University of Michigan,
1600 Huron Parkway, Ann Arbor, Michigan 48109 (USA)
E-mail: shaomeng@umich.edu
Prof. Dr. J. Stuckey, Dr. K, Chinnaswamy
Life Sciences Institute, University of Michigan, 210 Washtenaw, Ann
Arbor, Michigan 48109 (USA)
Dr. E. Fernandez-Salas
Department of Pathology, University of Michigan, 1600 Huron
Parkway, Ann Arbor, Michigan 48109 (USA)
[+]
These authors contributed equally to this work
Analysis of the co-crystal structure of MIV-6 complexed with
menin shows that there is a well-defined pocket around the
Supporting information for this article is given via a link at the end of
the document.
This article is protected by copyright. All rights reserved.

10.1002/anie.201711828
Angewandte Chemie International Edition
COMMUNICATION
cyclopentyl group of MIV-6[4c]. We reasoned that addition of an
appropriate group to the cyclopentyl ring to capture additional
interactions at this site could greatly enhance the binding affinity
to menin. Extensive modifications at this site showed that
installation of a reverse carbamate at the 2-position of the
cyclopentyl ring yielded compound 4 which showed dramatically
improved binding affinity to menin. Compound 4 binds to menin
with an IC50 value of 29 nM, and is >100-times more potent than
compounds 2 or 3.
suggesting that the nitrile group needs to be extended if it is to
form a covalent bond with Cys329 (Supplementary Information -
SI - Figure S1a). We thus replaced the nitrile group in 6 with a
cyclobutylsulfonyl group, yielding 7. Modeling suggests that the
sulfonyl group can maintain the hydrogen bond with the indole
group of Trp321 and the cyclobutyl moiety can lodge in the
surface hydrophobic pocket formed by Cys329, Val367, Val371
and Trp341 (SI Figure S1b). Indeed, it was found that 7 binds to
menin with IC50 = 7 nM (Ki < 1 nM).
Compound 4 was initially synthesized as a racemic compound.
To determine its stereospecificity for binding to menin, we
developed an efficient synthetic route to compound 5, a pure
enantiomer of 4. Compound 5 binds to menin with an IC50 value
of 15 nM, and is twice as potent as 4.
We next designed and synthesized 8 as a potential covalent
inhibitor by installation of an acrylamide group, a Michael acceptor,
on the 4-membered ring of 7. We also synthesized 9 as a
reversible inhibitor control by reducing the ethenyl moiety in 8 to
an ethyl group.
Introduction of a fluorine substituent into the free phenyl ring
of previously examined menin inhibitors[4c, 6] had been shown to
improve the binding affinity to menin and accordingly, we
systematically investigated the effect of a fluorine substituent in
the free phenyl group of 5. This effort resulted in 6, which binds to
menin with an IC50 value of 9 nM (Ki < 1 nM).
To establish the mode of action of 8 and 9, we performed
mass spectroscopic analysis (SI Table S1), which showed that 8
forms a covalent complex upon incubation with recombinant
human menin protein but 9, lacking the acrylamide, the Michael
acceptor group, fails to do so. Using bio-layer interferometry (BLI),
we demonstrated that while 8 binds to menin protein irreversibly,
9 binds to menin protein reversibly (SI Figure S2). These data
clearly show that 8 is a covalent, irreversible menin inhibitor, while
9 is a non-covalent, reversible menin inhibitor.
In the cell growth inhibition assay, the irreversible menin
inhibitor 8 achieves an IC50 value of 6 nM in the MV4;11 cell line,
and its corresponding reversible inhibitors, 7 and 9, have IC50
values of 210 and 263 nM respectively (Table 1). Hence, 8 is >30-
times more potent than the corresponding reversible inhibitors 7
and 9. In the cell growth inhibition assay, 8 has an IC50 value of
3.2 µM in the HL-60 cell line lacking MLL fusion, thus
displaying >500-times cellular selectivity in the MV4;11 cell line
harboring an MLL fusion over the HL-60 cell line lacking MLL
fusion.
Table 1. Binding affinity and cell growth inhibition of menin inhibitors.
Cell growth inhibition assay (4 days)
Binding Affinity to
menin in FP Assay
(IC50, nM)
(IC50, µM)
ID
MV4;11
HL60
(MLL fusion)
(No MLL fusion)
1 (MIV-6)
185 ± 24
3.6 ± 0.2
4.9 ± 0.8
2
3100 ± 200
3000 ± 200
29 ± 2
2.8 ± 0.32
> 10
> 10
> 10
3
4
0.83 ± 0.04
0.20 ± 0.01
0.16 ± 0.03
0.21 ± 0.02
0.006 ± 0.002
0.263 ± 0.039
0.0027 ± 0.0005
1.01 ± 0.14
> 10
5
15 ± 1
> 10
6
9.1 ± 1.0
6.9 ± 1.1
3.0 ± 1.0
8.0 ± 1.0
3.3 ± 0.4
> 2000
> 10
7
> 10
8
3.2 ± 0.7
> 10
To improve the solubility of compound 8, we attached a
dimethylaminomethyl group to the acrylamide group, yielding
compound 10, which binds to menin with an IC50 value of 3 nM.
In the cell growth inhibition assay, 10 achieves an IC50 value of 3
nM in the MV4;11 cell line and has an IC50 value of 2.0 µM in the
HL-60 cell line, again displaying >500-times cellular selectivity for
the MV4;11 cell line over the HL-60 cell line.
9
10 (M-525)
11
2.0 ± 0.4
1.5 ± 0.3
These high-affinity menin inhibitors (4-6) were tested for their
activity and selectivity in a 4-day cell growth inhibition assay in
MV4;11 (MLL-AF4) and HL-60 (no MLL fusion) leukemia cell lines
and the resulting data are summarized in Table 1. Compounds 4,
5 and 6 achieve IC50 values of 0.83, 0.20, and 0.16 µM,
respectively, in the MV4;11 cell line. Significantly, these three
compounds have IC50 values of >10 µM in the HL-60 cell line,
demonstrating excellent cellular selectivity for MLL leukemia.
Although compounds 5 and 6 bind to menin with very high
affinities (Ki < 1 nM), they achieve only submicromolar IC50 values
in the MV4;11 cell line carrying an MLL fusion. We next explored
the possibility of designing covalent, irreversible menin inhibitors
based upon the potent reversible inhibitor 6. Our goal was a
dramatic improvement of the cellular potency of menin inhibitors
through covalent and irreversible inhibition of menin.
Analysis of the co-crystal structure of the MIV-6/menin
complex shows that the Cys329 residue in menin is in close
proximity to the inhibitor molecule and may be used in the design
of covalent menin inhibitors. Our modeled structure of 6 in a
complex with menin shows that the terminal nitrile group of 6 is
the closest to the sulfur atom of Cys329 but is still 6.7 Å away,
Figure 2. Co-crystal structure of compound 10 (green) complexed with menin
(gray) at 2.61 Å resolution (PDB: 6B41). A covalent bond was found between
Cys329 and inhibitor 10.
This article is protected by copyright. All rights reserved.

10.1002/anie.201711828
Angewandte Chemie International Edition
COMMUNICATION
To establish precisely the mode of interaction of 10 with menin,
we determined the co-crystal structure of 10 complexed with
human menin protein at 2.61 Å resolution (Figure 2, SI Figure S3).
This co-crystal structure clearly shows that, consistent with our
design, 10 through its acrylamide group forms a covalent bond
with the sulfur atom of Cys329. The reverse carbamate group on
the cyclopentyl ring inserts nicely into the available pocket in
menin, its carbonyl group forms a strong hydrogen bond with the
hydroxyl group of Tyr276, and the methyl group enjoys
hydrophobic contacts with the side chains of Met278 and Cys241.
The fluorine substituent on the phenyl ring inserts into a
hydrophobic pocket formed by Met228, Met278 and Leu177. This
co-crystal structure provides a solid structural basis for the high-
affinity, covalent binding of 10 with menin. The mass
spectroscopic and BLI data further confirm that 10 is a covalent
and irreversible inhibitor (SI, Table S1 and Figure S2).
To investigate the binding stereospecificity of 10 with menin,
we synthesized 11, a diastereoisomer of 10, by inverting the chiral
center on the quaternary carbon (Figure 1). Compound 11 binds
to menin with an IC50 value of >2000 nM, and is thus >500-times
less potent than 10, indicating the high binding stereospecificity of
10 to menin. Furthermore, 11 has an IC50 value of 1.0 µM in
inhibition of cell growth of the MV4;11 cell line, and is >300-times
less potent than 10. These data are further evidence that the
potent cellular activity achieved by 10 in the MV4;11 cell line is
attributable to its binding to menin.
The cellular thermal shift assay (CETSA) is a powerful assay
allowing assessment of the engagement of small-molecule
inhibitors with their specific protein target in cells[7]. Our CETSA
data (SI Figure S4) demonstrated that 10 enhances the thermal
stability of cellular menin protein in both the MV4;11 and MOLM-
13 cells with respect to the control treatment at concentrations as
low as 0.4-1.2 nM, reaching a maximum effect at 30-100 nM. In
comparison, 10 has no effect on WD repeat-containing protein 5
(WDR5)[8], which also binds to MLL. Since the MV4;11 and
MOLM-13 cells were treated with 10 for only 1 h, the thermal
stabilization of cellular menin protein by 10 is evidently a direct
effect. Hence, the CETSA data provide clear evidence that 10
effectively targets the cellular menin protein at subnanomolar
concentrations.
1 2 0
1 0 0
8 0
6 0
4 0
2 0
0
M V 4 ;1 1 (M L L -A F 4 ) (IC 5 0
= 2.3 nM ))
M O L M -1 3 (M L L -A F 9 ) (IC 5 0
M O L M -1 4 (M L L -A F 9 ) (IC 5 0
=
=
1 5.4 nM )
3 1.3 nM )
S E M (M L L -A F 4 ) (IC 5 0
=
1 8.0 nM )
9.4 nM )
R S 4 ;1 1 (M L L -A F 4 ) (IC 5 0
=
M o n o M a c6 (M L L -A F 9 ) (IC 5 0
K 5 6 2 (n o M L L fu sio n ) (IC 5 0
=
1 0.3 nM )
=
4 .5  M )
M O L M -1 6 (n o M L L fu sio n ) (IC 5 0
= 6 .5  M )
S K M -1 (n o M L L fu sio n ) (IC 5 0
H L -6 0 (n o M L L fu sio n ) (IC 5 0
=
0 .9  M )
1 .6  M )
=
0.01
0.0001
0.001
0.1
1
10
[M -5 25 ] ( M )
Figure 3. Cell growth inhibition in acute leukemia cell lines with or without MLL
fusion. Cells were treated with compound 10 for 7 days and cell viability was
determined using a WST-8 assay. Data are representative of three independent
experiments.
Reversible menin inhibitors have been shown to down-
regulate MEIS1 and HOX genes in MV4;11 and other leukemia
cell lines carrying an MLL fusion[4d]. We assessed the ability of
compound 10 to regulate expression of MEIS1 and HOX genes in
the MV4;11 and MOLM-13 cell lines. In the MV4;11 cell line,
treatment for 6 or 24 h with 10 significantly down-regulates the
expression of MEIS1, HOXA9 and HOXA11 genes in a dose-
dependent manner (SI, Figure S5a and S5b) and is effective at
low nanomolar concentrations. Compound 10 also effectively
suppresses the expression of MEIS1 and HOX genes in the
MOLM-13 cell line at concentrations as low as 3 nM (SI, Figure
S5c). Compound 10 is therefore potent and effective in
suppressing the expression of MEIS1 and HOX genes in MV4;11
cells carrying the MLL-AF4 fusion and in MOLM-13 cells carrying
the MLL-AF9 fusion.
We further examined in
a
pharmacodynamics (PD)
experiment (SI, Figure 6a, b), the ability of 10 to down-regulate
MEIS1 and HOX genes in the MV4;11 xenograft tumor tissue in
mice. In contrast to the requirement for repeated dosing with
reversible menin inhibitors[4d], a single, intravenous administration
of 10 effectively down-regulates MEIS1, HOXA9 and HOXA11
genes after 24 h with the effect persisting for at least 48 h. The
long-lasting PD effect of a single dose of 10 on down-regulation
of MEIS1 and HOX genes in vivo suggests that infrequent
administration of an irreversible menin inhibitor may be sufficient
to achieve a strong anti-leukemia activity against MLL leukemia
in patients, and this would represent a significant advantage over
reversible menin inhibitors.
The synthesis of 10 is summarized in Scheme 1. The critical
step was the construction of the key intermediate (15a), which
contains three chiral centers and a highly hindered quaternary
carbon with four distinct substituents. The synthesis of 15a was
achieved using an efficient method that generates the three chiral
centers in one step. The absolute stereochemistry of 15a was
confirmed by determination of its single crystal structure. The
detailed syntheses of 10 and other compounds are provided in SI.
In summary, we report the succesful design and extensive
evaluation of first-in-class, irreversible inhibitors of menin,
examplified by 10 (M-525). Our data clearly demonstrate that
irreversible inhibitors of menin are highly potent and effective in
targeting the menin-MLL interaction and should be extensively
evaluated as a new class of targeted therapy for MLL leukemia.
The activity and specificity of compound 10 were further
evaluated in a panel of human acute leukemia cell lines with or
without MLL fusions. Similar to inhibitors of other epigenetic
regulators such as EZH2 and DOT1L[9], menin inhibitors exert
their cell growth inhibitory activity more slowly than traditional
cytotoxic agents and kinase inhibitors[4d]
. Accordingly, we
evaluated the growth inhibitory activity of 10 in a panel of leukemia
cell lines with a 7-day treatment, and obtained the results
summarized in Figure 3. Our data show that 10 achieves IC50
values of 2.3-31.3 nM in six leukemia cell lines harboring MLL
fusions. In comparison, 10 has IC50 values of 0.9-6.5 µM in four
leukemia cell lines lacking MLL fusion where it is 25-2000 times
less potent than in leukemia cell lines carrying an MLL fusion.
Thus, 10 achieves potent cellular activity in human leukemia cell
lines carrying MLL fusions, independent of the MLL fusion partner,
and demonstrates excellent cellular selectivity over leukemia cell
lines lacking MLL fusion.
This article is protected by copyright. All rights reserved.

10.1002/anie.201711828
Angewandte Chemie International Edition
COMMUNICATION
N. Kinukawa, A. Morimoto, K. Isoyama, Y. Kosaka, T. Oda, M. Oda, Y.
Hayashi, M. Eguchi, K. Horibe, T. Nakahata, S. Mizutani, E. Ishii,
Leukemia 2007, 21, 2258-2263.
[2]
a) C. Caslini, Z. Yang, M. El-Osta, T. A. Milne, R. K. Slany, J. L. Hess,
Cancer Res. 2007, 67, 7275-7283; b) Y.-X. Chen, J. Yan, K. Keeshan,
A. T. Tubbs, H. Wang, A. Silva, E. J. Brown, J. L. Hess, W. S. Pear, X.
Hua, Proc. Natl. Acad. Sci. U.S.A. 2006, 103, 1018-1023; c) A.
Yokoyama, T. C. Somervaille, K. S. Smith, O. Rozenblatt-Rosen, M.
Meyerson, M. L. Cleary, Cell 2005, 123, 207-218; d) R. K. Slany,
Haematologica 2009, 94, 984-993; e) Y. Zhang, A. Chen, X. M. Yan, G.
Huang, Int. J. Hematol. 2012, 96, 428-437.
[3]
[4]
a) A. Yokoyama, T. C. P. Somervaille, K. S. Smith, O. Rozenblatt-Rosen,
M. Meyerson, M. L. Cleary, Cell 2005, 123, 207-218; b) A. Yokoyama, M.
L. Cleary, Cancer Cell 2008, 14, 36-46.
Scheme 1. Reagents and conditions: a) MeONa, MeOH, reflux; b) NaBH4,
MeOH, rt; c) KHMDS, 18-Crown-6, H2SO4, THF, 0 oC to rt; d) TFA, CH2Cl2, 0 oC
to rt; e) Dimethyl dicarbonate, Et3N, CH2Cl2, 0 oC to rt; f) 10% Pd/C, H2, MeOH,
rt; g) K2CO3, KI, CH3CN, 80 oC; h) TFA, CH2Cl2, 0 oC to rt; i) DIPEA, CH2Cl2, 0
oC to rt. The absolute stereochemistry of 15a was confirmed by x-ray
crystallography (CDCC: 1581872).
a) J. Grembecka, S. He, A. Shi, T. Purohit, A. G. Muntean, R. J.
Sorenson, Nat. Chem. Biol. 2012, 8, 277-284; b) A. Shi, M. J. Murai, S.
He, G. Lund, T. Hartley, T. Purohit, Blood 2012, 120, 4461-4469; c) S.
He, T. J. Senter, J. Pollock, C. Han, S. K. Upadhyay, T. Purohit, R. D.
Gogliotti, C. W. Lindsley, T. Cierpicki, S. R. Stauffer, J. Grembecka, J.
Med. Chem. 2014, 57, 1543-1556; d) D. Borkin, S. He, H. Miao, K.
Kempinska, J. Pollock, J. Chase, T. Purohit, B. Malik, T. Zhao, J. Wang,
B. Wen, H. Zong, M. Jones, G. Danet-Desnoyers, Monica L. Guzman, M.
Talpaz, Dale L. Bixby, D. Sun, Jay L. Hess, Andrew G. Muntean, I.
Maillard, T. Cierpicki, J. Grembecka, Cancer Cell 2015, 27, 589-602.
J. Singh, R. C. Petter, T. A. Baillie, A. Whitty, Nat. Rev. Drug Discov.
2011, 10, 307-317.
Acknowledgements
This work is supported in part by the Prostate Cancer Foundation
and the National Cancer Institute, NIH (P30 CA046592, University
of Michigan Comprehensive Cancer Center Support Grant). Use
of the Advanced Photon Source, an Office of Science User
Facility operated for the U.S. Department of Energy (DOE) Office
of Science by Argonne National Laboratory, was supported by the
U.S. DOE under Contract No. DE-AC02-06CH11357. Use of the
LS-CAT Sector 21 was supported by the Michigan Economic
Development Corporation and the Michigan Technology Tri-
Corridor (Grant 085P1000817).
[5]
[6]
[7]
H. Zhou, L. Liu, J. Huang, D. Bernard, H. Karatas, A. Navarro, M. Lei, S.
Wang, J. Med. Chem. 2013, 56, 1113-1123.
a) D. M. Molina, R. Jafari, M. Ignatushchenko, T. Seki, E. A. Larsson, C.
Dan, L. Sreekumar, Y. Cao, P. Nordlund, Science 2013, 341, 84-87; b)
R. Jafari, H. Almqvist, H. Axelsson, M. Ignatushchenko, T. Lundbäck, P.
Nordlund, D. M. Molina, Nat. Protoc. 2014, 9, 2100-2122.
[8]
[9]
J.-J. Song, R. E. Kingston, J. Biol. Chem. 2008, 283, 35258-35264.
a) S. K. Knutson, N. M. Warholic, T. J. Wigle, C. R. Klaus, C. J. Allain, A.
Raimondi, M. Porter Scott, R. Chesworth, M. P. Moyer, R. A. Copeland,
V. M. Richon, R. M. Pollock, K. W. Kuntz, H. Keilhack, Proc. Natl. Acad.
Sci. U.S.A. 2013, 110, 7922-7927; b) S. R. Daigle, E. J. Olhava, C. A.
Therkelsen, A. Basavapathruni, L. Jin, P. A. Boriack-Sjodin, C. J. Allain,
C. R. Klaus, A. Raimondi, M. P. Scott, N. J. Waters, R. Chesworth, M. P.
Moyer, R. A. Copeland, V. M. Richon, R. M. Pollock, Blood 2013, 122,
1017-1025.
Keywords: drug design • irreversible inhibitors • menin-MLL
protein-protein interaction• MLL leukemia
[1]
a) P. Ernst, J. Wang, S. J. Korsmeyer, Curr. Opin. Hematol. 2002, 9, 282-
287; b) R. Marschalek, Br. J. Haematol. 2011, 152, 141-154; c) R.
Popovic, N. J. Zeleznik-Le, J. Cell. Biochem. 2005, 95, 234-242; d) R. K.
Slany, Hematol. Oncol. 2005, 23, 1-9; e) D. Tomizawa, K. Koh, T. Sato,
This article is protected by copyright. All rights reserved.

10.1002/anie.201711828
Angewandte Chemie International Edition
COMMUNICATION
COMMUNICATION
The irreversible inhibitor M-525
targets the menin-MLL protein-protein
interaction. Our study demonstrates
that irreversible inhibition of menin
represents a promising therapeutic
strategy for the treatment of MLL
leukemia and may have advantages
over reversible inhibition.
S. Xu, A. Aguilar, T. Xu, K. Zheng, L.
Huang, J. Stuckey, K, Chinnaswamy, D.
Bernard, E. Fernández-Salas, L. Liu, M.
Wang, D. McEachern, S. Przybranowski,
C. Foster, S. Wang*
Page No. – Page No.
A First-in-Class, Highly Potent Irreversible
Inhibitor Targeting the Menin-MLL Protein-
Protein Interaction
This article is protected by copyright. All rights reserved.