Inhibitors Targeting STAT5 Signaling in Myeloid Leukemias: New Tetrahydroquinoline Derivatives with Improved Antileukemic Potential

Article abstract of DOI:10.1002/cmdc.202000841

Signal transducers and activators of transcription 5A and 5B (STAT5A and STAT5B) are two closely related STAT family members that are crucial downstream effectors of tyrosine kinase oncoproteins such as FLT3-ITD in acute myeloid leukemia (AML) and BCR-ABL

Full text of DOI:10.1002/cmdc.202000841

Accepted Article
Title: Inhibitors Targeting STAT5 Signaling in Myeloid Leukemias: New
Tetrahydroquinoline Derivatives with Improved Antileukemic
Potential
Authors: Gildas Prié, Marion Polomski, Marie Brachet-Botineau,
Ludovic Juen, Marie-Claude Viaud-Massuard, and Fabrice
Gouilleux
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To be cited as: ChemMedChem 10.1002/cmdc.202000841
Link to VoR: https://doi.org/10.1002/cmdc.202000841
01/2020

10.1002/cmdc.202000841
ChemMedChem
FULL PAPER
Inhibitors Targeting STAT5 Signaling in Myeloid Leukemias: New
Tetrahydroquinoline Derivatives with Improved Antileukemic
Potential
Marion Polomski,[a],# Marie Brachet-Botineau,[b],# Ludovic Juen,[a] Marie-Claude Viaud-Massuard,[a]
Fabrice Gouilleux[b] and Gildas Prié [a],
*
[a]
M. Polomski, Dr. L. Juen, Prof. M.-C. Viaud-Massuard, Dr. G. Prie
Equipe IMT « Innovation Moléculaire et Thérapeutique » - GICC EA7501
Université de Tours - Labex SYNORG
Faculté de Pharmacie - 31 av Monge - 37200 Tours - France
E-mail: gildas.prie@univ-tours.fr
[b]
#
Dr. M. Brachet-Botineau, Dr. F. Gouilleux
Equipe LNOx « Niche Leucémique & Métabolisme Oxydatif » - GICC ERL 7001 CNRS
Université de Tours
Faculté de Médecine - Bâtiment Dutrochet - 10bis bvd Tonnellé - 37032 Tours - France
Author Contributions, M.P. and M.B.-B. contributed equally
Supporting information for this article is given via a link at the end of the document.
Abstract: Signal transducers and activators of transcription 5A and
5B (STAT5A and STAT5B) are two closely related STAT family
members that are crucial downstream effectors of tyrosine kinase
oncoproteins such as FLT3-ITD in acute myeloid leukemia (AML)
and BCR-ABL in chronic myeloid leukemia (CML). We recently
developed and reported the synthesis of a first molecule called 17f
that selectively inhibits STAT5 signaling in myeloid leukemia cells
and overcomes their resistance to chemotherapeutic agents. To
improve the antileukemic effect of 17f, we synthesized 10 analogs of
this molecule and analyzed their impact on cell growth, survival,
totally curative and is unable to completely eradicate leukemic
stem cells (LSC) which is responsible of relapse of CML patients.
Moreover, mutations on BCR-ABL tyrosine kinase domain
occurs, which prevents IM binding and promote IM resistance of
CML cells.[11–15]
Mutations affecting the tyrosine kinase receptor FLT3 can
be found in up to one third of AML patients. FLT3-ITD is the
most frequent one and usually confers a poor prognosis.[16]
Targeting the kinase activity of FLT3 mutants has been therefore
adopted to inhibit aberrant signaling in AML. First- and second-
chemoresistance and STAT5 signaling. Two compounds, 7a and 7a’, generations FLT3 inhibitors were employed as a single agent or
were identified as having similar or higher antileukemic effects in
various AML and CML cell lines. Both molecules were found to be
more effective than 17f to inhibit STAT5 activity/expression and to
suppress the chemoresistance of CML.
in combination with other therapeutic drugs such as cytarabine
(Ara-C) and anthracyclines in AML treatment.[17–20] Clinical
activity of some of these molecules was evidenced in patients
with FLT3-mutated AML but was often transient and relapse
eventually occurred. Like BCR-ABL, secondary mutations in
FLT3 have been shown to be responsible for the loss of
therapeutic response to FLT3 inhibitors.[21] Alternative
approaches targeting critical effectors of BCR-ABL and FLT3-
ITD are therefore needed to cure CML and AML.
Introduction
Among the STAT family members, STAT5A and STAT5B
proteins play a critical role in the development of hematologic
cancers that affect lymphoid and myeloid lineages.[1] Persistent
tyrosine phosphorylation of STAT5A/5B (P-Y-STAT5) has been
found in hematopoietic cancers driven by tyrosine kinase
oncogenes (TKO) such as BCR-ABL in chronic myeloid and
acute lymphoid leukemias (CML and Ph+ALL), FLT3-internal
tandem duplication (FLT3-ITD) in acute myeloid leukemia (AML)
and JAK2V617F in myeloproliferative neoplasms (MPN).[2–4] In
addition, gain of function (GOF) mutations of STAT5B have
been described in different hematopoietic malignancies affecting
the lymphoid compartment. This is best illustrated by the
presence of the recurrent N642H mutation of STAT5B in some T
cell leukemias.[5]
In this regard, STAT5 satisfy all criteria to be a druggable
target in CML and AML diseases.[22,23] STAT5 proteins have
been shown to play a crucial role in the maintenance of CML
stem cells and to promote the resistance of CML cells against
TKI treatments.[24–27] Expression of P-Y-STAT5 has also been
associated with an increased risk of relapse in AML after
achieving complete remission.[28] Thus, targeting STAT5 would
contribute to abrogate de novo and acquired resistance of CML
and AML to chemotherapy.
Several approaches have been used to target STAT5 in
leukemias.[23] Among them, chromones- and salicylic acid-based
molecules have been employed to identify selective inhibitors
that bind to the SH2 domain of STAT5.[29–31] One of them, AC-4-
130 suppressed P-Y-STAT5 and the growth of CML and AML
cells as well as tumor formation in xenograft models of AML.[32,33]
Finally, another small STAT5-SH2-binding compound containing
The development of Imatinib Mesylate (IM, Glivec®) and
related BCR-ABL kinase inhibitors such as Dasatinib, Nilotinib,
Bosutinib and Ponatinib has made a major breakthrough in the
treatment of CML.[6–10] IM leads to complete inhibition of BCR-
ABL-dependent STAT5 activation which is an important part of
this molecule effectiveness. However, Imatinib Mesylate is not
a nucleotidic scaffold showing
a
therapeutic potential for
prostate cancer and CML has been identified in structure-based
screening studies.[34] In a similar vein, cell-based screening
assays using a library of clinically used drugs were also
1
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10.1002/cmdc.202000841
ChemMedChem
FULL PAPER
employed to identify the antipsychotic drug, pimozide, as a Chemistry
potential STAT5 inhibitor. This compound was shown to inhibit
P-Y-STAT5 and survival of CML and AML cells without affecting
the kinase activity of BCR-ABL or FLT3-ITD.[35,36] Synthesis of
pimozide derivatives and structure-activity relationship (SAR)
studies allowed to identify compounds with enhanced inhibitory
effects on STAT5 activity and CML cell growth.[37] As
repositioning drugs, antidiabetics such as pioglitazone and
rosiglitazone have also been shown to possess antileukemic
activity. Both synthetic compounds belong to the
thiazolidinedione (TZD) class of ligands that bind to the nuclear
receptor peroxisome proliferator-activated receptor gamma
(PPARγ). Activation of PPARγ by pioglitazone not only inhibits
the growth of CML cells but also reduces the expression of
STAT5 genes.[38–40] Importantly, the drug combination using
pioglitazone with IM triggers apoptosis of these leukemic cells
suggesting that besides phosphorylation, inhibition of STAT5
expression is of prime importance for resistant CML stem cell
eradication.[40]
In order to study SAR around the tetrahydroquinoline
(THQ) ring, we used a convergent pathway implemented and
optimized from our previously described work. This synthetic
route, setting good yields, has been extended to synthesize a
series of new analogs using the brominated THQ and indole
rings as starting materials with the linking strategy as described
previously (Scheme 1).[41]
Scheme 1. Our work on new analogs
We used a library of PPAR agonists that were previously
synthesized in our lab, to identify a hit compound that blocks P-
Y-STAT5 and leukemic cell growth. 18 derivatives of this “hit”
To obtain these analogs series, we needed to synthetize
the brominated key intermediates 4,4-dimethyl-1,2,3,4-
tetrahydroquinoline 6, with bromine on positions 5, 7 and 8.
From 2-bromoaniline, we performed the same method used in
Juen et al. work with an acylation followed by a Friedel-Crafts
alkylation to get lactam 2d.[41] From 3-bromoaniline, we obtained
as expected both lactam brominated on positions 5 and 7, and
isolated each isomer by purification, respectively 2a and 2c.
Each lactam was then reduced to the corresponding brominated
THQ 3a,c,d. Alcohol 4 was converted to aldehyde by 2-
iodobenzoic acid (IBX) and then filtered on amines 3a and 3c in
the presence of NaBH(OAc)3, to afford by reductive amination
5a and 5c. From amine 3d, compound 5d was not observed,
surely due to steric hindrance of the bromine in position 8 on the
tetrahydroquinoline scaffold. Consequently, an alternative
synthesis pathway from amine 3d has been developed to obtain
the desired 8-brominated THQ. Finally, deprotection of 5a and
5c using potassium tert-butoxide under air (1 atm) in DMSO led
to key intermediates 6a and 6c, with 68% and 74% yields
respectively (Scheme 2).
sharing
a
diversely
substituted
4,4-dimethyl-1,2,3,4-
tetrahydroquinoline scaffold, linked to an heteroaryl ring by an
ethoxy chain were synthesized and tested in preclinical studies
using various leukemic cell lines. Results of the cell proliferation
and viability assays allowed us to select one compound called
17f with improved antileukemic activity. This compound is
composed of a 3-pyridinyl substituted 4,4-dimethyl-1,2,3,4-
tetrahydroquinoline (THQ) and indole rings, linked by an ethoxy
chain attached by carbon C-5 of the indole and nitrogen N-1 of
the THQ (Figure 1). We showed that 17f suppressed P-Y-
STAT5 and growth of CML and AML cells.[41] We also
demonstrated that 17f sensitizes leukemic cells that acquired
resistance to IM or Ara-C.[42]
Figure 1. Compound 17f
In order to improve the antileukemic potential of 17f, we
synthesized 10 new analogs with modifications around the
tetrahydroquinoline (THQ) ring and analyzed their impacts on
CML and AML cell growth. Among these analogs, two
compounds, 7a and 7a’, containing respectively a 3- and 4-
pyridinyl moiety in position 5 on the THQ ring, showed similar or
higher growth inhibitory effects when compared to 17f. We also
demonstrated that 7a and 7a’ were more effective than 17f in
blocking P-Y-STAT5 expression and activity. Lastly, both
compounds showed a slightly better cytotoxic effect when
combined with IM to kill CML cells that acquired resistance to
this tyrosine kinase inhibitor.
Scheme 2. Synthesis of key intermediates 6a-c. Reagents and conditions: (i)
3,3-dimethylacryloyl chloride, dry pyridine, 0 °C to rt, 5 h; (ii) AlCl3, dry DCM,
0 °C to rt, 1 h; (iii) BH3.THF, dry toluene, 0 °C to reflux, 5 h; (iv) 1) alcohol 4,
IBX, dry DCE, 80 °C, 2 h; 2) filtration; 3) amine 3, NaBH(OAc)3, rt, 3 h; (v) t-
BuOK, DMSO, O2, rt, 3 h.
2
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Introduction of 3- and 4-pyridinyl groups on the THQ ring of
both 6a-c by Suzuki cross-coupling afforded respectively 7a-c
and 7a’-c’ analogs with satisfying yields (40-61%) (Scheme 3).
We then focused on analogs substituted in position 8 on the
THQ ring. Indeed, as previously described, no reaction between
amine 3d and alcohol 4 was observed due to the bromine
position. Therefore, Suzuki cross-coupling with 3- and 4-pyridinyl
boronic acids were performed first on brominated THQ 3d to
afford 11d,d’. Thereafter classic pathway was used to reach
compounds 13d,d’ in two steps and unsurprisingly, the bridge
formation first step gave low yields (Scheme 5).
Scheme 3. Synthesis of analogs 7a-c and 7a’-c’ by Suzuki coupling from
compounds 6a-c. Reagents and conditions: R1B(OH)2, PdCl2(PPh3)2, K2CO3,
DMF/water, 90 °C, 5 h.
However, as observed by Juen et al., the reaction between
6a-c and 2-pyridinylboronic acid did not occur and only led to
degradation of the starting material. To bypass this issue, we
converted aryl bromides 6a-c into pinacol boronate esters 8a-c
with good yields (50-68%). Compounds 8b and 8c subsequently
yielded 9b and 9c after Suzuki coupling using 2-
bromopyridine.[43,44] Concerning compound 9b, this optimized
procedure allowed us to improve the yield from 24% to 60% in
two steps.[41] Though, concerning pinacol boronate ester 8a,
coupling was not observed, surely due to steric hindrance
between methyl groups of both pinacol boronate and THQ.
Consequently, we synthesized the corresponding less hindered
neopentyl boronate ester 10 from aryl bromide 6a and used it in
the same way in Suzuki coupling with 2-bromopyridine to yield
9a with a satisfying 71% yield (Scheme 4).
Scheme 5. Synthesis of analogs substituted in position
8 on the
tetrahydroquinoline ring. Reagents and conditions: (i) R1B(OH)2, PdCl2(PPh3)2,
K2CO3, DMF/water, 90 °C, 5 h; (ii) 1) alcohol 4, IBX, dry DCE, 80 °C, 2 h; 2)
filtration; 3) amine 12, NaBH(OAc)3, rt, 3 h; (iii) t-BuOK, DMSO, O2, rt, 3 h.
Introduction of 2-pyridinyl ring on position 8 was a double
challenge, considering known difficulties with 2-pyridinyl
coupling reactions and THQ position 8 hindrance, so several
strategies were implemented. Previously described, formation of
pinacol or neopentyl boronate ester were tested on aryl bromide
3d, in order to perform Suzuki coupling with 2-bromopyridine.
However, none of these reactions occurred certainly due to
interactions between the free amine hydrogen and the bromine.
To avoid this, we decided to protect the amine function by a
benzyl group. Two strategies were employed starting from
compound 3d: reductive amination using benzaldehyde in the
presence of NaBH(OAc)3 and classical N-alkylation from
benzylbromide using sodium hydride. Unfortunatly, none of the
two reactions yielded the desired protected amine. Two different
strategies of complete synthesis were then considered starting
from
(Supplementary
2-aminophenylboronic
acid
or
route
2-chloroaniline
using 2-
data).
Synthetic
aminophenylboronic acid permitted to obtain the THQ
substituted with a 2-pyridine group in position 8, however all
attempts to yield the compound linked to indole by the ethyloxy
chain failed. We explained this by the steric hindrance with the
2-pyridinyl group in this 8-position and thus by possible
interaction between the hydrogen of amine function on
tetrahydroquinoline scaffold and the 2-pyridine nitrogen. From 2-
chloroaniline, we achieved synthesis of linked compound with
chloride in position 8 on tetrahydroquinoline ring with a very low
5% yield. Then, deprotected compound was obtained and used
to form the corresponding pinacol boronate or neopentyl
boronate ester but all reactions failed and led to the starting
chlorinated material.
Scheme 4. Synthesis of 2-pyridinyl analogs from compounds 6a-c. Reagents
and conditions: (i) bis(pinacolato)diboron, PdCl2(dppf), KOAc, dry dioxane,
100 °C, 12 h; (ii) bis(neopentylglycolato)diboron, PdCl2(dppf), KOAc, dry
dioxane, 100 °C, 12 h; (iii) 2-bromopyridine, PdCl2(dppf), Na2CO3,
dioxane/water, 100 °C, 5 h.
3
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Results and discussion
on the THQ ring might directly impact the antileukemic potential
of 17f.
Initial screenings were carried out to determine IC50 values
for newly synthesized compounds tested on CML (KU812 and
K562) and AML (MV4-11) cell lines (Table 1).
In order to improve the antileukemic activity of 17f, SAR
studies were conducted with focus on the pyridinyl heterocycle
bound to the tetrahydroquinoline ring. In this work, we then
addressed whether changes in the nitrogen position on the
pyridinyl moiety as well as changes in the position of the latter
Table 1. Structure of analogs 7a-c and 7a’-c’
IC50 (µM)
Compound
CML
AML
Substitution
on THQ ring
R
KU812
K562
MV4-11
17f = 7b
6a
6
5
6
7
5
5
6
7
7
6
7
5
6
7
5
8
8
3-py
Br
5.38 ± 0.01
10.74 ± 3.07
7.59 ± 0.63
30.9 ± 8.27
5.80 ± 0.80
2.50 ± 0.10
9.38 ± 1.18
84.45 ± 19.60
93.21 ± 8.19
35.23 ± 11.78
3.15 ± 1.18
16.00 ± 4.19
>200
8.74 ± 1.54
17.17 ± 3.33
15.58 ± 1.67
3.55 ± 0.47
27.89 ± 2.91
7.14 ± 0.85
4.86 ± 1.25
5.90 ± 0.70
2.80 ± 0.40
13.35 ± 2.83
25.30 ± 11.32
945 ± 151.20
23.56 ± 4.28
14.88 ± 4.80
14.88 ± 5.43
15.25 ± 1.82
88.15 ± 14.33
6.57 ± 1.99
4.25 ± 1.51
6.85 ± 0.34
6b
Br
6c
Br
55.13 ± 10.70
3.12 ± 0.50
5.20 ± 0.70
22.65 ± 1,09
103.70 ± 13.30
444 ± 6.02
7a
3-py
4-py
4-py
3-py
4-py
Bpin
Bpin
2-py
2-py
2-py
Bneop
3-py
4-py
7a’
7b’
7c
7c’
8b
21.46 ± 5.28
15.50 ± 1.14
51.06 ± 10.77
>900
8c
9a
9b
9c
89.50 ± 4.51
41.23 ± 4.54
21.40 ± 6.02
20.25 ± 6.04
914.16 ± 195.30
91.43 ± 2.07
22.6 ± 7.35
15.5 ± 4.99
10
13d
13d’
aIC50 values of 17f analogs and their precursors obtained for K562, KU812 and MV4-11 cell lines. Cells were treated with concentrations ranging from 100 nM to
100 µM for 48 h. Cytotoxicity was determined by MTT assays. IC50 values were calculated using GraphPad Prism 7 software (n = 3 in triplicate, mean ± SEM).
Compounds 7c, 7c’ and 13d, 13d’ substituted respectively in
position 7 and 8 on the THQ ring were less effective in
inhibiting leukemic cell growth than 17f. In addition, results
clearly indicated that substitution by the 2-pyridinyl group in
each position on the THQ ring (analogs 9a-c) dramatically
reduced the effectiveness of 17f. Precursor compounds 6a-c,
8b,c and 10 showed average IC50 values similar or higher
than 17f on both CML and AML cell lines. In sharp contrast,
compounds 7a and 7a’ substituted in position 5 on THQ ring
respectively by the 3- and 4-pyridinyl moieties showed similar
or enhanced antileukemic activity when compared to 17f.
Thereby, activity of compounds 7a, 7a’ and 17f seems to be
promoted by their pyridinyl nitrogen similar position (i.e.
possible formation of a hydrogen bond in a same area). This
observation can explain also the lower activity of compound
7b', where pyridinyl nitrogen is slightly offset from this area.
To confirm these results in other leukemic cells, IC50 values of
7a and 7a’ were also determined in KCL-22 (CML), KG1a and
MOLM-13 (AML) cell lines. Excepted for KG1a cells, IC50
values obtained for 7a and 7a’ were nearly close to those
obtained for 17f (Table 2A). Besides, compounds 6c, 10, 13d
and 13d’, which only showed a good activity on MV4-11 cells,
were also evaluated on KG1a and MOLM-13 cell lines but did
not exhibit relevant results (Supplementary data). Additionally,
4
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cytotoxicity effects of 7a and 7a’ were also determined on
human bone marrow stromal cells (cell line: HS-27a and
primary mesenchymal stem cells: MSC) which play a crucial
role in the leukemic niche. Results showed that cytotoxic
effects of 7a and 7a’ on HS-27a and MSC were slightly higher
than those observed for 17f but IC50 values remained 5 to 10-
fold higher than those required to inhibit leukemic cell growth
(Table 2B).
Table 2. Cytotoxic effects of 7a, 7a’ and 17f in CML, AML and human bone
marrow stromal cells
A
IC50 (µM)
CML
Compound
AML
MOLM-13
KCL-22
5.6 ± 0.8
6.9 ± 3.1
8.7 ± 1.9
KG1a
17f
7a
3.4 ± 0.9
7.8 ± 0.9
6.9 ± 0.8
3.3 ± 1.0
5.4 ± 0.5
4.7 ± 1.0
7a’
B
Toxicity on Stromal Cells - IC50 (µM)
Compound
HS-27a
MSC
17f
7a
63.7 ± 0.1
27.0 ± 1.4
23.2 ± 5.0
33.6 ± 5.9
27.1 ± 1.4
12.5 ± 0.5
Figure 2. (A) Viability kinetics of KU812 and MV4-11 treated with
compounds 17f, 7a and 7a’. Cells were treated with 5 µM of inhibitors or
with DMSO as control, for 24, 48 or 72 h. Cytotoxic effect was determined
by MTT assays. Results are presented in percentage of control. Values
were calculated using GraphPad Prism 7 software (n = 3 in triplicate, mean
± SEM). (B) Growth kinetics of KU812 (up) and MV4-11 (down) treated
with 5 µM of compounds 17f, 7a and 7a’ or DMSO as control determined
by Trypan blue exclusion assays (n = 3 in triplicate, mean ± SEM). (C)
Representative flow cytometry histogram of KU812 and MV4-11 cells
treated with 3 µM of 7a, 7a’ and 17f or DMSO. Cells were stained with
FITC Annexin V and 7-AAD, and the percentage of apoptotic cells were
then evaluated by flow cytometry (n = 3 in triplicate; mean ± SEM).
7a’
a(A) IC50 values of selected compounds obtained for KCL-22, KG1a and
MOLM-13 cell lines. (B) IC50 values of selected compounds obtained for
HS-27a and MSC stromal cells. (A-B) Cells were treated with
concentrations ranging from 100 nM to 100 µM for 48 h and cytotoxicity
was determined by MTT assays. IC50 values were calculated using
GraphPad Prism 7 software (n = 3 in triplicate, mean ± SEM).
To analyze the growth inhibitory properties of 7a and
7a’, KU812 and MV4-11 cells were treated or not (DMSO as
control) with 5µM (concentration that roughly corresponds to
that of the 17f IC50 values) of 7a, 7a’ and 17f during 72h. Cell
viability and the number of living cells were daily determined
by MTT and trypan blue dye exclusion assays respectively.
Results showed that 7a and 7a’, when compared to 17f, have
a slightly higher or similar cytotoxic effect in KU812 and MV4-
11 cells respectively (Figure 2A). Both compounds have also
similar or enhanced growth inhibitory properties (Figure 2B)
and were able to induce apoptosis in both leukemic cell lines
(Figure 2C).
We previously demonstrated that 17f inhibits STAT5
signaling in CML and AML cells, we then addressed whether
7a and 7a’ also suppress STAT5 activity and expression in
KU812 and MV4-11 cells. A concentration of 3 µM was used
in the experiments to keep close to the IC50 values of both
compounds. We first evaluated impact of 7a and 7a’ on
STAT5A and STAT5B gene expression in both leukemic cell
lines by qRT-PCR experiments. Results showed that 7a and
7a’ had no effect on STAT5A and STAT5B mRNA levels in
leukemic cells treated for 15 h with these compounds (Figure
3A). We then analyzed the phosphorylation and expression of
STAT5 in KU812 and MV4-11 cells exposed to 7a and 7a’
(3µM) for 24 and 48 h. Results showed that both compounds
inhibited the phosphorylation of STAT5 and reduced STAT5
protein expression at 48 h post-treatment suggesting that 7a
and 7a’ as 17f might indirectly interfere with the
stability/degradation of the proteins (Figure 3B).[42] Further
5
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investigations are then required to determine impact of these
molecules on the ubiquitin/proteasome system.
overcome the resistance of CML cells to IM. K562R cells that
were previously used as an IM-resistant leukemic cell model,
were treated with various concentrations of 17f, 7a and 7a’
(ranging from 0.1 to 10 µM) in combination with 1 µM IM.
Results showed that compound 7a’ exhibited a slightly but
significant better cytotoxic effect than 17f and 7a at 1 and 5
µM concentrations (figure 5A). This was also confirmed in
kinetic experiments using 5 µM as the difference in the
optimal concentration between each compound (figure 5B).
Figure 5. (A) Metabolic activity of IM-resistant cells K562 R treated with IM
(1 µM) in combination with ranging doses from 100 nM to 10 µM of 7a, 7a’
or 17f, for 48 h. Cytotoxic effect was determined by MTT assays. Results
are presented in percentage of control. Values were calculated using
GraphPad Prism 7 software (n = 3 in triplicate, mean ± SEM). (B) Viability
kinetics of IM-resistant cells K562 R treated with IM (1 µM) as control in
combination with selected compound, 17f, 7a or 7a’ (5 µM) for 24, 48 or 72
h. Cytotoxic effect was determined by MTT assays. Results are presented
in percentage of control. Values were calculated using GraphPad Prism 7
software (n = 3 in triplicate; mean ± SEM).
Figure 3. (A) qRT-PCR analysis of STAT5A and STAT5B transcripts in
KU812 and MV4-11 cells treated or not with 7a, 7a’ or DMSO as control for
24 h. Results are presented as the fold changes in gene expression in
treated cells normalized to internal control genes (GAPDH, ACTB,
RPL13a) and relative to control cells (normalized to 1) (n = 3 in triplicate;
mean ± SEM). (B) Protein extracts from KU812, MV4-11 cells treated with
7a and 7a’ (3 µM) or DMSO (−) for 48 h were analyzed by Western blotting
to detect P-Y-STAT5 and STAT5 protein expression. Actin was used as a
loading control (n = 3).
Conclusion
In summary, among the 10 new analogs of 17f,
compounds 7a and 7a’ were found to be the most interesting
inhibitors of STAT5 signaling and myeloid leukemia cell
We then addressed whether 7a and 7a’ inhibited STAT5
transcriptional activity. qRT-PCR experiments were
conducted to determine effects of these compounds on
STAT5-dependent expression of target genes such as PIM1
and CISH in KU812 and MV4-11 cells. 17f was used as a
positive control in these experiments. We found that both
molecules were as efficient as 17f to downregulate PIM1 and
CISH gene expression (Figure 4).
growth when compared to 17f.
Our SAR studies
demonstrated that both positions of pyridinyl moiety on the
THQ aromatic part and its nitrogen position were essential in
the antileukemic properties of 7a and 7a’. Indeed, the C-5
tetrahydroquinoline 3- and 4- pyridinyl substitutions in 7a and
7a’, gave better IC50 results than C-7 and C-8 THQ ring
substitutions as well as all 2-pyridinyl substituted derivatives.
This work also showed that, as 17f, both 7a and 7a’ block
STAT5 transcriptional activity by affecting not only the
phosphorylation but also expression of the proteins.
Experiments are now conducted to determine whether both
compounds can affect expression of STAT5A, STAT5B and
most importantly the oncogenic STAT5BN642H mutant which is
frequently expressed in different T cell leukemias/lymphomas
and in T cell lymphoblastic leukemia.[45,46] SAR studies will be
then continued on our compound central scaffold. In particular
substitutions in positions C-5 and C-6 of the THQ ring by
polynitrogen aromatic heterocycles like pyrimidine or pyrazine
will be conducted to generate molecules with improved
antileukemic potential while avoiding their cytotoxic effects on
Figure 4. qRT-PCR analysis of STAT5 target genes, PIM1 and CISH
transcripts, in KU812 (left) and MV4-11 (right) cells treated or not with 7a,
7a’, 17f (3 μM) or DMSO as the control for 24 h. Results are presented as
the fold changes in gene expression in treated cells normalized to internal
control genes (GAPDH, ACTB, RPL13a) and relative to control cells
(normalized to 1) (n = 3 in triplicate; mean ± SEM).
normal cells. This should help us to get
a better
understanding of the influence of nitrogen on the THQ part of
our structure.
We previously showed that 17f overcomes the
resistance of CML and AML cells to IM and Ara-C.[42] We then
ask whether 7a and 7a’ had a greater impact than 17f to
Experimental Section
6
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10.1002/cmdc.202000841
ChemMedChem
FULL PAPER
CHEMISTRY
title compound 1b as a white solid (26.70 g, quant.); mp 115 °C. 1H
NMR (300 MHz, DMSO-d6): δ 10.01 (s, 1H), 8.02 (dd, J = 1.8, 1.8 Hz,
1H), 7.48 (ddd, J = 7.8, 1.8, 1.8 Hz, 1H), 7.25 (dd, J = 7.8, 7.8 Hz, 1H),
7.19 (ddd, J = 7.8, 1.8, 1.8 Hz, 1H), 5.84 (dq, J = 1.1, 1.1 Hz, 1H),
2.15 (d, J = 1.1 Hz, 3H), 1.87 (d, J = 1.1 Hz, 3H). 13C NMR (75 MHz,
DMSO-d6): δ 164.8, 152.4, 141.2, 130.7, 125.4, 121.5, 121.2, 118.8,
117.7, 27.1, 19.6. HRMS-ESI (m/z): found 254.01729, cacld for
C11H1379BrNO+ [M+H]+ 254.01750 and found 256.01488, cacld for
C11H1381BrNO+ [M+H]+ 256.01546.
Materials and methods
Reagents and solvents were purchased from commercial sources
(Acros, Aldrich and VWR international) and were used without further
purification. For synthesis in anhydrous and inert conditions, the
glassware was dried with a heat gun during several vacuum-dry argon
cycles and maintained under argon. Reactions were monitored by
using thin layer chromatography (TLC), pre-coated aluminum sheets
silica gel 60 F254 marketed by Merck and visualized under UV
fluorescence 254 and 365 nm. Purifications were carried out by
chromatography on silica gel columns on an ISCO purification unit,
Combi Flash RF 75 PSI, with Redisep flash silica gel columns (60 Å,
230-400 mesh, grade 9385). Melting point (mp) were determined on a
Büchi melting point apparatus Model B-540 in open capillaries. NMR
spectra were recorded at 300 MHz (1H) or 75 MHz (13C) on a Bruker
AVANCE AV 300 instrument. All NMR experiments were realized in
deuterated chloroform (CDCl3) or dimethylsulfoxide (DMSO-d6).
Chemical shifts were reported in parts per million (ppm) (δ relative to
residual solvent peak for 1H and 13C) and coupling constants (J) are
reported in hertz (Hz). The notations used are: δ: chemical shift
(ppm); s: singlet; d: doublet; dd: doublet of doublet; t: triplet; m:
multiplet; br: broad signal; J: coupling constant (Hz). High-resolution
mass spectra (HRMS) was performed by the mass spectrometry
service on a Q-Exactive spectrometer from Thermo Scientific using
the electrospray ionization (ESI) technique. If necessary, the purity
was determined by high performance liquid chromatography (HPLC).
Purity of all final compounds was 95% or higher. HPLC analyses were
carried out with a LaChrom Elite system [Hitachi L-2130 (pump), L-
2200 (autosampler) and L-2400 (UV-detector)] using 254 nm UV for
detection. The column was a XBridge C-18 (250 × 10 mm, 4μm,
135Å); elution was performed with 0.1% (by volume) of TFA in water
(solvent A), and 0.1% of TFA in acetonitrile (solvent B); gradient 20-
100% of B over 35 minutes with a flow rate of 1 mL.min-1; column
temperature of 25 °C; injection of 10 µL in DMSO.
Method B: Cyclization (2a, 2c, 2d).
To a solution of N-bromophenyl-3-methylbut-2-enamide 1 (1.0 equiv)
in dry dichloromethane at 0 °C under argon was added portionwise
aluminium chloride (4.0 equiv) over 30 min. The reaction mixture was
stirred for 1 h at rt, then poured on ice and extracted with diethyl ether.
After extraction, the combined organic extracts were washed with
saturated aqueous NaHCO3 solution, brine and then dried with
MgSO4. After evaporation under reduced pressure, the crude product
was purified by flash chromatography on silica gel.
2a 5-Bromo-4,4-dimethyl-3,4-dihydroquinolin-2(1H)-one and 2c 7-
bromo-4,4-dimethyl-3,4-dihydroquinolin-2(1H)-one
Following method B, the reaction of N-(3-bromophenyl)-3-methylbut-
2-enamide 1b (20.00 g; 78.70 mmol) and aluminium chloride (42.00 g,
314.80 mmol) in dry dichloromethane (97 mL) afforded, after
purification by flash chromatography on silica gel (cyclohexane/ethyl
acetate; 70/30), the title compounds 2a as a white solid (7.38 g, 43 %)
and 2c as a white solid (9.05 g, 47 %).
2a: mp 137 °C. 1H NMR (300 MHz, DMSO-d6): δ 10.28 (brs, 1H), 7.20
(dd, J = 7.9, 1.3 Hz, 1H), 7.04 (d, J = 7.9, 7.9 Hz, 1H), 6.88 (dd, J =
7.9, 1.3 Hz, 1H), 2.43 (s, 2H), 1.44 (s, 6H). 13C NMR (75 MHz, DMSO-
d6): δ 168.4, 139.8, 130.0, 128.9, 128.6, 120.7, 115.8, 46.8, 36.5, 27.0
(2C). HRMS-ESI (m/z): found 254.01728, cacld for C11H1379BrNO+
[M+H]+ 254.01750 and found 256.01501, cacld for C11H1381BrNO+
[M+H]+ 256.01546.
Method A: Nucleophilic substitution (1a, 1b).
To a solution of bromoaniline (1.0 equiv) in dry pyridine at 0 °C under
argon, was added dropwise 3,3-dimethylacryloylchloride (1.2 equiv).
The reaction mixture was stirred for 5 h at rt and then carefully poured
into an Erlenmeyer flask containing 1 M HCl under magnetic stirring.
The product was filtered, washed by water and heptane, then purified
by flash chromatography on silica gel.
2c: mp 175 °C. 1H NMR (300 MHz, DMSO-d6) δ 10.25 (brs, 1H), 7.23
(d, J = 8.3 Hz, 1H), 7.13 (dd, J = 8.3, 2.0 Hz, 1H), 7.03 (d, J = 2.0 Hz,
1H), 2.36 (s, 2H), 1.20 (s, 6H). 13C NMR (75 MHz, DMSO-d6) δ 169.4,
138.7, 131.5, 126.5, 124.8, 119.5, 117.6, 44.5, 33.4, 27.0 (2C).
HRMS-ESI (m/z): found 254.01706, cacld for C11H1379BrNO+ [M+H]+
254.01750 and found 256.01466, cacld for C11H1381BrNO+ [M+H]+
256.01546.
1a N-(2-Bromophenyl)-3-methylbut-2-enamide
Following method A, the reaction of 2-bromoaniline (4.60 mL, 40.70
mmol), 3,3-dimethylacryloyl chloride (5.50 mL, 48.84 mmol) in dry
pyridine (17.70 mL) afforded, after purification by flash
chromatography on silica gel (cyclohexane/ethyl acetate; 70/30), the
title compound 1a as a white solid (10.30 g, quant.); mp 100 °C. 1H
NMR (300 MHz, DMSO-d6): δ 9.28 (s, 1H), 7.68 – 7.58 (m, 2H), 7.40
– 7.30 (m, 1H), 7.15 – 7.07 (m, 1H), 6.05 – 5.96 (m, 1H), 2.13 (d, J =
1.1 Hz, 3H), 1.86 (d, J = 1.0 Hz, 3H). 13C NMR (75 MHz, DMSO-d6): δ
164.6, 152.1, 136.5, 132.6, 127.8, 127.1, 126.7, 118.6, 117.9, 27.0,
19.6. HRMS-ESI (m/z): found 254.01704, cacld for C11H1379BrNO+
[M+H]+ 254.01750 and found 256.01461, cacld for C11H1381BrNO+
[M+H]+ 256.01546.
2d 8-Bromo-4,4-dimethyl-3,4-dihydroquinolin-2(1H)-one
Following method B, the reaction of N-(2-bromophenyl)-3-methylbut-
2-enamide 1a (20.00 g; 78.70 mmol) and aluminium chloride (42.00 g,
314.80 mmol) in dry dichloromethane (97 mL) afforded, after
purification by flash chromatography on silica gel (cyclohexane/ethyl
acetate; 70/30), the title compound 2d as a white solid (18.85 g, 94%);
mp 108 °C. 1H NMR (300 MHz, DMSO-d6): δ 9.17 (s, 1H), 7.46 (dd, J
= 7.8, 1.2 Hz, 1H), 7.33 (dd, J = 7.8, 1.2 Hz, 1H), 6.96 (dd, J = 7.8, 7.8
Hz, 1H), 2.41 (s, 2H), 1.24 (s, 6H).13C NMR (75 MHz, DMSO-d6): δ
169.4, 135.5, 134.7, 130.9, 124.1, 123.9, 109.7, 44.6, 34.3, 26.9 (2C).
HRMS-ESI (m/z): found 254.01710, cacld for C11H1379BrNO+ [M+H]+
254.01750 and found 256.01492, cacld for C11H1381BrNO+ [M+H]+
256.01546.
1b N-(3-Bromophenyl)-3-methylbut-2-enamide
Following method A, the reaction of 3-bromoaniline (11.40 mL,
105.00 mmol), 3,3-dimethylacryloyl chloride (14.00 mL, 126.00 mmol)
in dry pyridine (46.00 mL) afforded, after purification by flash
chromatography on silica gel (cyclohexane/ethyl acetate; 70/30), the
Method C: Reduction (3a, 3c, 3d).
7
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10.1002/cmdc.202000841
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FULL PAPER
To a solution of bromo-4,4-dimethyl-3,4-dihydroquinolin-2(1H)-one 2
(1.0 equiv) in dry toluene at 0 °C under argon was added dropwise a
solution of BH3.THF 1 M (2.5 equiv). The reaction mixture was
refluxed for 4 h and then quenched at 0 °C by the addition of
saturated aqueous NaHCO3 solution. After extraction with ethyl
acetate, the combined organic extracts were washed with brine and
dried with MgSO4. After evaporation under reduced pressure, the
crude product was purified by flash chromatography on silica gel.
MgSO4. After evaporation under reduced pressure, the crude product
was purified by flash chromatography on silica gel.
5a
1-(2-((1-Benzyl-1H-indol-5-yl)oxy)ethyl)-5-bromo-4,4-dimethyl-
1,2,3,4-tetrahydroquinoline
Following method D, the reaction of 2-((1-benzyl-1H-indol-5-
yl)oxy)ethanol 4 (0.35 g, 1.32 mmol), IBX (1.11 g, 3.96 mmol), 5-
bromo-4,4-dimethyl-1,2,3,4-tetrahydroquinoline 3a (0.66 g, 2.77
mmol), NaBH(OAc)3 (0.56 g, 2.64 mmol) in dry 1,2-DCE (5 mL)
afforded, after purification by flash chromatography on silica gel
(cyclohexane/ethyl acetate; 80/20), a mixture of title compound 5a
with starting material 3a. The mixture was dissolved in diethyl ether
(200 mL), washed by HCl 3 M (3 × 100 mL) and brine (1 × 100 mL),
dried with MgSO4 and evaporated under reduced pressure to give 5a
as a light yellow oil (0.51 g, 78%). 1H NMR (300 MHz, DMSO-d6): δ
7.44 (d, J = 3.1 Hz, 1H), 7.33 – 7.20 (m, 4H), 7.17 – 7.12 (m, 2H),
7.04 (d, J = 2.3 Hz, 1H), 6.88 – 6.69 (m, 4H), 6.35 (d, J = 3.1 Hz, 1H),
5.36 (s, 2H), 4.10 (t, J = 5.7 Hz, 2H), 3.68 (t, J = 5.7 Hz, 2H), 3.34 –
3.29 (m, 2H), 1.77 – 1.68 (m, 2H), 1.46 (s, 6H). 13C NMR (75 MHz,
DMSO-d6): δ 153.0, 147.5, 138.8, 131.6, 130.1, 129.2, 128.9 (2C),
128.3, 128.2, 127.7, 127.3 (2C), 123.6, 122.8, 112.0, 111.9, 111.3,
103.7, 101.0, 65.5, 51.9, 49.7, 46.0, 41.3, 34.1, 28.8 (2C). HRMS-ESI
(m/z): found 489.15329, cacld for C28H3079BrN+ [M+H]+ 489.15360 and
found 491.15117, cacld for C28H3081BrN+ [M+H]+ 491.15156.
3a 5-Bromo-4,4-dimethyl-1,2,3,4-tetrahydroquinoline
Following method C, the reaction of 5-bromo-4,4-dimethyl-3,4-
dihydroquinolin-2(1H)-one 2a (3.00 g; 11.80 mmol) and BH3.THF 1 M
(30.00 mL, 29.51 mmol) in dry toluene (28 mL) afforded, after
purification by flash chromatography on silica gel (cyclohexane/ethyl
acetate; 80/20), the title compound 3a as a light yellow oil (2.28 g,
81%). 1H NMR (300 MHz, CDCl3): δ 6.91 (dd, J = 7.8, 1.4 Hz, 1H),
6.75 (dd, J = 8.0, 7.8 Hz, 1H), 6.45 (dd, J = 8.0, 1.4 Hz, 1H), 3.78 (brs,
1H), 3.24 – 3.16 (m, 2H), 1.85 – 1.78 (m, 2H), 1.57 (s, 6H). 13C NMR
(75 MHz, CDCl3): δ 146.7, 128.1, 127.6, 124.7, 124.1, 114.9, 41.9,
38.0, 33.9, 28.7 (2C). HRMS-ESI (m/z): found 240.03797, cacld for
C11H1579BrN+ [M+H]+ 240.03824 and found 242.03542, cacld for
C11H1581BrN+ [M+H]+ 242.03619.
3c 7-Bromo-4,4-dimethyl-1,2,3,4-tetrahydroquinoline
5c
1-(2-((1-Benzyl-1H-indol-5-yl)oxy)ethyl)-7-bromo-4,4-dimethyl-
Following method C, the reaction of 7-bromo-4,4-dimethyl-3,4-
dihydroquinolin-2(1H)-one 2c (7.00 g; 27.50 mmol) and BH3.THF 1 M
(70.00 mL, 69.00 mmol) in dry toluene (65 mL) afforded, after
purification by flash chromatography on silica gel (cyclohexane/ethyl
acetate; 80/20), the title compound 3c as a light yellow oil (5.95 g,
90%). 1H NMR (300 MHz, CDCl3): δ 7.03 (d, J = 8.2 Hz, 1H), 6.74 (dd,
J = 8.2, 2.1 Hz, 1H), 6.62 (d, J = 2.1 Hz, 1H), 3.87 (brs, 1H), 3.34 –
3.28 (m, 2H), 1.75 – 1.69 (m, 2H), 1.27 (s, 6H). 13C NMR (75 MHz,
CDCl3): δ 144.8, 129.3, 128.0, 120.0, 119.9, 116.6, 38.4, 36.8, 31.7,
30.8 (2C). HRMS-ESI (m/z): found 240.03807, cacld for C11H1579BrN+
[M+H]+ 240.03824 and found 242.03541, cacld for C11H1581BrN+
[M+H]+ 242.03619.
1,2,3,4-tetrahydroquinoline
Following method D, the reaction of 2-((1-benzyl-1H-indol-5-
yl)oxy)ethanol 4 (0.50 g, 1.87 mmol), IBX (1.57 g, 5.61 mmol), 7-
bromo-4,4-dimethyl-1,2,3,4-tetrahydroquinoline 3c (0.95 g, 3.92
mmol), NaBH(OAc)3 (0.80 g, 3.74 mmol) in dry 1,2-DCE (7 mL)
afforded, after purification by flash chromatography on silica gel
(cyclohexane/ethyl acetate; 98/2), a mixture of title compound 5c with
starting material 3c. The mixture was dissolved in diethyl ether (200
mL), washed by HCl 3 M (3 × 100 mL) and brine (1 × 100 mL), dried
with MgSO4 and evaporated under reduced pressure to give 5c as a
light yellow oil (0.45 g, 49%). 1H NMR (300 MHz, DMSO-d6): δ 7.44 (d,
J = 3.1 Hz, 1H), 7.34 – 7.21 (m, 4H), 7.17 – 7.12 (m, 2H), 7.06 (d, J =
2.4 Hz, 1H), 7.04 (d, J = 8.2 Hz, 1H), 6.80 (d, J = 1.9 Hz, 1H), 6.71
(dd, J = 8.9, 2.4 Hz, 1H), 6.62 (dd, J = 8.2, 1.9 Hz, 1H), 6.36 (d, J =
3.1 Hz, 1H), 5.36 (s, 2H), 4.12 (t, J = 5.4 Hz, 2H), 3.67 (t, J = 5.4 Hz,
2H), 3.45 – 3.39 (m, 2H), 1.66 – 1.60 (m, 2H), 1.17 (s, 6H). 13C NMR
(75 MHz, DMSO-d6): δ 152.5, 145.2, 138.4, 131.1, 129.8, 129.7,
128.7, 128.5 (2C), 127.4, 127.3, 126.9 (2C), 119.8, 117.4, 112.6,
111.5, 110.9, 103.2, 100.6, 65.1, 50.1, 49.2, 45.9, 35.9, 31.4, 30.0
(2C). HRMS-ESI (m/z): found 489.15317, cacld for C28H3079BrN+
[M+H]+ 489.15360 and found 491.15109, cacld for C28H3081BrN+
[M+H]+ 491.15156.
3d 8-Bromo-4,4-dimethyl-1,2,3,4-tetrahydroquinoline
Following method C, the reaction of 8-bromo-4,4-dimethyl-3,4-
dihydroquinolin-2(1H)-one 2d (10.00 g; 39.30 mmol) and BH3.THF 1
M (100.00 mL, 98.20 mmol) in dry toluene (92 mL) afforded, after
purification by flash chromatography on silica gel (cyclohexane/ethyl
acetate; 80/20), the title compound 3d as a light yellow oil (8.96 g,
76%). 1H NMR (300 MHz, CDCl3): δ 7.26 (dd, J = 7.9, 1.4 Hz, 1H),
7.17 (dd, J = 7.7, 1.4 Hz, 1H), 6.53 (dd, J = 7.9, 7.7 Hz, 1H), 4.69 (brs,
1H), 3.49 – 3.36 (m, 2H), 1.83 – 1.71 (m, 2H), 1.32 (s, 6H). 13C NMR
(75 MHz, CDCl3): δ 140.5, 132.0, 130.1, 125.4, 117.2, 109.0, 38.5,
36.7, 32.4, 30.9 (2C). HRMS-ESI (m/z): found 240.03775, cacld for
C11H1579BrN+ [M+H]+ 240.03824 and found 242.03521, cacld for
C11H1581BrN+ [M+H]+ 242.03619.
Method E: Heterocyclic amine debenzylation (6a, 6c, 13d-d’).
To a solution of the benzyl indole 5 or 12 (1.0 equiv) in DMSO at rt
was added t-BuOK (14.0 equiv). Dry air was then bubbled into the
solution and stirred 2 h at rt. The reaction mixture was quenched with
saturated aqueous NH4Cl solution, extracted with ethyl acetate,
washed with brine and dried with MgSO4. After evaporation under
reduced pressure, the crude product was purified by flash
chromatography on silica gel.
Method D: Reductive Amination (5a, 5c, 12a, 12b).
To a solution of 2-((1-benzyl-1H-indol-5-yl)oxy)ethanol 4 (1.0 equiv) in
dry 1,2-DCE under argon, was added 2-iodoxybenzoic acid (IBX) (3.0
equiv). The resulting mixture was stirred 2 h at 80 °C and then cooled
to rt. The reaction medium was filtered through a sintered glass funnel
over a round bottom flask containing the corresponding amine (2.1
equiv) in dry 1,2-DCE. The resulting mixture was stirred for 15
minutes and then NaBH(OAc)3 (2.0 equiv) was added. The reaction
medium was stirred 2 h at rt and then hydrolyzed with a saturated
aqueous NaHCO3 solution. After extraction with ethyl acetate, the
combined organic extracts were washed with brine and dried with
6a
1-(2-((1H-Indol-5-yl)oxy)ethyl)-5-bromo-4,4-dimethyl-1,2,3,4-
tetrahydroquinoline
Following method E, the reaction of 1-(2-((1-benzyl-1H-indol-5-
yl)oxy)ethyl)-5-bromo-4,4-dimethyl-1,2,3,4-tetrahydroquinoline
5a
(2.02 g; 4.13 mmol) and t-BuOK (6.50 g , 57.78 mmol) in DMSO
8
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10.1002/cmdc.202000841
ChemMedChem
FULL PAPER
(122.50 mL) afforded, after purification by flash chromatography on
Following method F, the reaction of 1-(2-((1H-indol-5-yl)oxy)ethyl)-5-
bromo-4,4-dimethyl-1,2,3,4-tetrahydroquinoline 6a (80 mg, 0.20
mmol), pyridin-4-ylboronic acid (74 mg, 0.60 mmol), K2CO3 (83 mg,
0.60 mmol) and PdCl2(PPh3)2 (14 mg, 0.02 mmol) in DMF (4.00 mL)
and water (0.80 mL) afforded, after purification by flash
chromatography on silica gel (cyclohexane/ethyl acetate; 70/30), the
title compound 7a’ as a yellow solid (41 mg, 51%); mp 180 °C. HPLC
purity: 99.8 %, tR = 16.65 min. 1H NMR (300 MHz, DMSO-d6): δ 10.92
(brs, 1H), 8.53 (d, J = 4.9 Hz, 2H), 7.31 – 7.24 (m, 4H), 7.05 (d, J =
2.4 Hz, 1H), 6.98 (dd, J = 8.4, 7.3 Hz, 1H), 6.80 – 6.76 (m, 1H), 6.73
(dd, J = 8.7, 2.4 Hz, 1H), 6.33 – 6.29 (m, 1H), 6.06 (dd, J = 7.3, 0.9 Hz,
1H), 4.18 (t, J = 5.6 Hz, 2H), 3.75 (t, J = 5.6 Hz, 2H), 3.51 – 3.43 (m,
2H), 1.62 – 1.54 (m, 2H), 0.95 (s, 6H). 13C NMR (75 MHz, DMSO-d6):
δ 152.4, 148.6 (2C), 144.4, 142.5, 139.5, 131.1, 128.0 (2C), 126.0,
125.8, 124.8 (2C), 118.4, 112.0, 111.5, 111.2, 102.7, 100.8, 65.0,
50.9, 45.7, 39.0, 32.9, 30.6 (2C). HRMS-ESI (m/z): found 398.21954,
cacld for C26H28N3O+ [M+H]+ 398.22269.
silica gel (cyclohexane/ethyl acetate; 80/20), the title compound 6a as
t
a light yellow oil (1.61 g, 98%). HPLC purity: 98.8%, R = 30.06 min.
1H NMR (300 MHz, DMSO-d6): δ 10.91 (brs, 1H), 7.28 – 7.23 (m, 2H),
7.02 (d, J = 2.4 Hz, 1H), 6.89 – 6.82 (m, 1H), 6.79 – 6.67 (m, 3H),
6.32 – 6.27 (m, 1H), 4.11 (t, J = 5.7 Hz, 2H), 3.69 (t, J = 5.7 Hz, 2H),
3.36 – 3.29 (m, 2H), 1.78 – 1.71 (m, 2H), 1.48 (s, 6H). 13C NMR (75
MHz, DMSO-d6): δ 152.3, 147.1, 131.1, 128.0, 127.9, 127.7, 125.8,
123.2, 122.3, 112.0, 111.5, 111.4, 102.7, 100.8, 65.0, 51.5, 45.6, 40.8,
33.6, 28.4 (2C). HRMS-ESI (m/z): found 399.10623, cacld for
C21H2479BrN2O+ [M+H]+ 399.10665 and found 401.10407, cacld for
C21H2481BrN2O+ [M+H]+ 401.10461.
6c
1-(2-((1H-Indol-5-yl)oxy)ethyl)-7-bromo-4,4-dimethyl-1,2,3,4-
tetrahydroquinoline
Following method E, the reaction of 1-(2-((1-benzyl-1H-indol-5-
yl)oxy)ethyl)-5-bromo-4,4-dimethyl-1,2,3,4-tetrahydroquinoline
5c
7c 1-(2-((1H-Indol-5-yl)oxy)ethyl)-4,4-dimethyl-7-(pyridin-3-yl)-1,2,3,4-
tetrahydroquinoline
(1.00 g; 2.04 mmol) and t-BuOK (3.21 g, 28.60 mmol) in DMSO (60
mL) afforded, after purification by flash chromatography on silica gel
(cyclohexane/ethyl acetate; 80/20), the title compound 6c as a yellow
t
1
oil (0.60 g, 74%). HPLC purity: 99.4%, R = 29.57 min. H NMR (300
MHz, DMSO-d6): δ 10.92 (brs, 1H), 7.31 – 7.23 (m, 2H), 7.09 – 7.01
(m, 2H), 6.82 (d, J = 2.0 Hz, 1H), 6.70 (dd, J = 8.7, 2.4 Hz, 1H), 6.63
(dd, J = 8.2, 2.0 Hz, 1H), 6.32 – 6.27 (m, 1H), 4.13 (t, J = 5.5 Hz, 2H),
3.68 (t, J = 5.5 Hz, 2H), 3.48 – 3.40 (m, 2H), 1.68 – 1.60 (m, 2H), 1.18
(s, 6H). 13C NMR (75 MHz, DMSO-d6): δ 152.3, 145.2, 131.1, 129.8,
128.0, 127.4, 125.8, 119.9, 117.4, 112.7, 112.0, 111.4, 102.7, 100.8,
65.1, 50.1, 45.9, 35.9, 31.4, 30.0 (2C). HRMS-ESI (m/z): found
399.10635, cacld for C21H2479BrN2O+ [M+H]+ 399.10665 and found
401.10420, cacld for C21H2481BrN2O+ [M+H]+ 401.10461.
Following method F, the reaction of 1-(2-((1H-indol-5-yl)oxy)ethyl)-7-
bromo-4,4-dimethyl-1,2,3,4-tetrahydroquinoline 6c (100 mg, 0.25
mmol), pyridin-3-ylboronic acid (92 mg, 0.75 mmol), K2CO3 (104 mg,
0.75 mmol) and PdCl2(PPh3)2 (17 mg, 0.025 mmol) in DMF (5.00 mL)
and water (1.20 mL) afforded, after purification by flash
chromatography on silica gel (cyclohexane/ethyl acetate; 60/40), the
title compound 7c as a yellow solid (60 mg, 61%); mp 209 °C. HPLC
purity: 99.4 %, tR = 18.14 min. 1H NMR (300 MHz, DMSO-d6): δ 10.90
(brs, 1H), 8.84 (d, J = 1.9 Hz, 1H), 8.52 (dd, J = 4.8, 1.3 Hz, 1H), 7.99
– 7.94 (m, 1H), 7.42 (dd, J = 7.9, 4.8 Hz, 1H), 7.28 – 7.21 (m, 3H),
7.06 (d, J = 2.4 Hz, 1H), 6.91 (d, J = 1.5 Hz, 1H), 6.82 (dd, J = 7.8, 1.5
Hz, 1H), 6.70 (dd, J = 8.7, 2.4 Hz, 1H), 6.31 – 6.25 (m, 1H), 4.20 (t, J
= 5.6 Hz, 2H), 3.80 (t, J = 5.6 Hz, 2H), 3.52 – 3.45 (m, 2H), 1.75 –
1.66 (m, 2H), 1.25 (s, 6H). 13C NMR (75 MHz, DMSO-d6): δ 152.3,
147.6, 147.3, 144.1, 136.7, 135.4, 134.3, 131.1, 130.7, 128.0, 126.5,
125.8, 123.8, 113.9, 112.0, 111.5, 109.0, 102.7, 100.8, 65.2, 50.2,
46.0, 36.4, 31.5, 30.4 (2C). HRMS-ESI (m/z): found 398.21970, cacld
for C26H27N3O+ [M+H]+ 398.22269.
Method F: Suzuki coupling (7a-a’, 7c-c’, 11d-d’)
A mixture of the corresponding aryl bromide 6 (1.0 equiv), K2CO3 (1.5
equiv) and the corresponding boronic acid (3.0 equiv) in DMF and
water was degassed with argon. PdCl2(PPh3)2 (0.1 equiv) was then
added and the resulting mixture was stirred 5 h at 90 °C. The mixture
was cooled to rt, filtered through a pad of dicalite and extracted with
ethyl acetate. Combined organic extracts were washed with brine and
dried with MgSO4. After evaporation under reduced pressure, the
crude product was purified by flash chromatography on silica gel.
7c’
1-(2-((1H-Indol-5-yl)oxy)ethyl)-4,4-dimethyl-7-(pyridin-4-yl)-
1,2,3,4-tetrahydroquinoline
Following method F, the reaction of 1-(2-((1H-indol-5-yl)oxy)ethyl)-7-
bromo-4,4-dimethyl-1,2,3,4-tetrahydroquinoline 6c (100 mg, 0.25
mmol), pyridin-4-ylboronic acid (92 mg, 0.75 mmol), K2CO3 (104 mg,
0.75 mmol) and PdCl2(PPh3)2 (17 mg, 0.025 mmol) in DMF (5 mL)
and water (1 mL) afforded, after purification by flash chromatography
on silica gel (cyclohexane/ethyl acetate; 60/40), the title compound
7c’ as a white solid (58.70 mg, 59%); mp: 209 °C. HPLC purity:
7a 1-(2-((1H-Indol-5-yl)oxy)ethyl)-4,4-dimethyl-5-(pyridin-3-yl)-1,2,3,4-
tetrahydroquinoline
Following method F, the reaction of 1-(2-((1H-indol-5-yl)oxy)ethyl)-5-
bromo-4,4-dimethyl-1,2,3,4-tetrahydroquinoline 6a (75 mg, 0.19
mmol), pyridin-3-ylboronic acid (69 mg, 0.56 mmol), K2CO3 (78 mg,
0.56 mmol) and PdCl2(PPh3)2 (13 mg, 0.02 mmol) in DMF (4.50 mL)
and water (0.90 mL) afforded, after purification by flash
chromatography on silica gel (cyclohexane/ethyl acetate; 70/30), the
title compound 7a as a yellow oil (30 mg, 40%). HPLC purity: 97.9%,
tR = 16.57 min. 1H NMR (300 MHz, DMSO-d6): δ 10.93 (brs, 1H), 8.52
(dd, J = 4.8, 1.7 Hz, 1H), 8.42 (d, J = 1.7 Hz, 1H), 7.63 (ddd, J = 7.8,
1.7, 1.7 Hz, 1H), 7.38 (dd, J = 7.8, 4.8 Hz, 1H), 7.31 – 7.25 (m, 2H),
7.06 (d, J = 2.4 Hz, 1H), 6.99 (dd, J = 8.5, 7.3 Hz, 1H), 6.78 (dd, J =
8.5, 1.3 Hz, 1H), 6.74 (dd, J = 8.7, 2.4 Hz, 1H), 6.34 – 6.28 (m, 1H),
6.13 (dd, J = 7.3, 1.3 Hz, 1H), 4.18 (t, J = 5.6 Hz, 2H), 3.75 (t, J = 5.6
t
99.9%, R = 18.31 min. 1H NMR (300 MHz, DMSO-d6): δ 10.92 (brs,
1H), 8.53 – 8.48 (m, 2H), 7.56 – 7.51 (m, 2H), 7.30 – 7.22 (m, 3H),
7.07 (d, J = 2.3 Hz, 1H), 6.95 (d, J = 1.5 Hz, 1H), 6.89 (dd, J = 7.9, 1.5
Hz, 1H), 6.71 (dd, J = 8.7, 2.3 Hz, 1H), 6.31 – 6.26 (m, 1H), 4.21 (t, J
= 5.5 Hz, 2H), 3.80 (t, J = 5.5 Hz, 2H), 3.52 – 3.44 (m, 2H), 1.74 –
1.68 (m, 1H), 1.25 (s, 6H).13C NMR (75 MHz, DMSO-d6): δ 152.3,
149.9 (2C), 148.0, 144.2, 135.6, 131.8, 131.1, 128.0, 126.5, 125.8,
121.1 (2C), 113.7, 112.0, 111.5, 108.7, 102.8, 100.8, 65.2, 50.1, 45.9,
36.3, 31.6, 30.3 (2C). HRMS-ESI (m/z): found 398.21947, cacld for
C26H27N3O+ [M+H]+ 398.22269.
Hz, 2H), 3.47 (t, J = 6.0 Hz, 2H), 1.62 – 1.53 (m, 2H), 0.90 (s, 6H). 13
C
NMR (75 MHz, DMSO-d6): δ 152.4, 149.3, 147.5, 144.5, 140.9, 138.4,
136.4, 131.1, 128.2, 128.0, 126.0, 125.8, 122.5, 119.5, 112.0, 111.5,
111.3, 102.7, 100.8, 65.0, 51.0, 45.7, 39.3, 32.9, 30.9, 30.7. HRMS-
ESI (m/z): found 398.21964, cacld for C26H27N3O+ [M+H]+ 398.22269.
Method G: Boronate formation (8a-c, 10)
A mixture of the corresponding aryl bromide 6 (1.0 equiv), KOAc (5.0
equiv) and the corresponding bisdiboron (1.2 equiv) in dioxane was
degassed with argon. Then PdCl2(dppf) (0.07 equiv) was added and
the resulting mixture was stirred at 100 °C for 12 h. The mixture was
cooled to rt, filtered through a pad of dicalite, washed with water and
7a’
1-(2-((1H-Indol-5-yl)oxy)ethyl)-4,4-dimethyl-5-(pyridin-4-yl)-
1,2,3,4-tetrahydroquinoline
9
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10.1002/cmdc.202000841
ChemMedChem
FULL PAPER
extracted with ethyl acetate. Combined organic extracts were washed
with brine and dried with MgSO4. After evaporation under reduced
pressure, the crude product was purified by flash chromatography on
silica gel.
Following method H, the reaction of 1-(2-((1H-indol-5-yl)oxy)ethyl)-5-
(5,5-dimethyl-1,3,2-dioxaborinan-2-yl)-4,4-dimethyl-1,2,3,4-
tetrahydroquinoline 10 (30 mg, 0.069 mmol), 2-bromopyridine (5.1 µL,
0.053 mmol), Na2CO3 (14 mg, 0.133 mmol) and PdCl2(dppf) (3 mg,
0.004 mmol) in dioxane (0.34 mL) and water (0.09 mL) afforded, after
purification by flash chromatography on silica gel (cyclohexane/ethyl
acetate; 60/40), the title compound 9a as a light yellow solid (15 mg,
8a 1-(2-((1H-Indol-5-yl)oxy)ethyl)-4,4-dimethyl-5-(4,4,5,5-tetramethyl-
1,3,2-dioxaborolan-2-yl)-1,2,3,4-tetrahydroquinoline
t
1
71%); mp : 56 °C. HPLC purity: 98.2%, R = 17.39 min. H NMR (300
MHz, DMSO-d6): δ 10.90 (s, 1H), 8.61 (d, J = 4.6 Hz, 1H), 7.83 – 7.74
(m, 2H), 7.41 – 7.35 (m, 1H), 7.32 – 7.17 (m, 5H), 7.06 (d, J = 1.9 Hz,
1H), 6.73 (dd, J = 8.7, 2.2 Hz, 1H), 6.32 – 6.25 (m, 1H), 4.21 (t, J =
5.8 Hz, 2H), 3.79 (t, J = 5.8 Hz, 2H), 3.52 – 3.44 (m, 2H), 1.75 – 1.68
(m, 2H), 1.25 (s, 6H). 13C NMR (75 MHz, DMSO-d6): δ 156.9, 152.3,
149.2, 143.9, 137.1, 136.8, 131.7, 131.1, 128.0, 126.1, 125.8, 122.0,
120.0, 113.9, 112.0, 111.5, 108.6, 102.8, 100.8, 65.0, 50.2, 45.9, 36.4,
31.6, 30.4 (2C). HRMS-ESI (m/z): found 398.22185, cacld for
C26H28N3O+ [M+H]+ 398.22269.
Following method G, the reaction of 1-(2-((1H-indol-5-yl)oxy)ethyl)-5-
bromo-4,4-dimethyl-1,2,3,4-tetrahydroquinoline 6a (335 mg, 0.84
mmol), bis(pinacolato)diboron (256 mg, 1.01 mmol), KOAc (412 mg,
4.20 mmol) and PdCl2(dppf) (43 mg, 0.059 mmol) in dioxane (5 mL)
afforded, after purification by flash chromatography on silica gel
(cyclohexane/ethyl acetate; 80/20), the title compound 8a as a light
yellow solid (188 mg, 50%).
8b 1-(2-((1H-Indol-5-yl)oxy)ethyl)-4,4-dimethyl-6-(4,4,5,5-tetramethyl-
1,3,2-dioxaborolan-2-yl)-1,2,3,4-tetrahydroquinoline
9b 1-(2-((1H-Indol-5-yl)oxy)ethyl)-4,4-dimethyl-6-(pyridin-2-yl)-1,2,3,4-
tetrahydroquinoline
Following method G, the reaction of 1-(2-((1H-indol-5-yl)oxy)ethyl)-6-
bromo-4,4-dimethyl-1,2,3,4-tetrahydroquinoline 6b (660 mg, 1.65
mmol), bis(pinacolato)diboron (504 mg, 1.98 mmol), KOAc (811 mg,
8.26 mmol) and PdCl2(dppf) (84.7 mg, 0.116 mmol) in dioxane (8.3
mL) afforded, after purification by flash chromatography on silica gel
(cyclohexane/ethyl acetate; 80/20), the title compound 8b as a beige
solid (468 mg, 63%); mp : 201 °C. HPLC purity: 95.2%, tR = 29.52 min.
1H NMR (300 MHz, DMSO-d6): δ 10.91 (s, 1H), 7.40 (d, J = 1.5 Hz,
1H), 7.32 – 7.22 (m, 3H), 7.02 (d, J = 2.3 Hz, 1H), 6.73 – 6.64 (m, 2H),
6.31 – 6.27 (m, 1H), 4.14 (t, J = 5.5 Hz, 2H), 3.72 (t, J = 5.5 Hz, 2H),
3.52 – 3.45 (m, 2H), 1.69 – 1.63 (m, 2H), 1.24 (s, 12H), 1.20 (s,
6H).13C NMR (75 MHz, DMSO-d6): δ 152.3, 146.1, 133.8, 131.6,
131.1, 129.5, 128.0, 125.8, 112.0, 111.4, 109.8, 102.7, 100.8, 82.8
(2C), 64.9, 49.9, 46.0, 36.2, 31.3, 30.2 (4C), 24.7 (2C). HRMS-ESI
(m/z): found 447.28125, cacld for C27H36BN2O3+ [M+H]+ 447.28135.
Following method H, the reaction of 1-(2-((1H-indol-5-yl)oxy)ethyl)-
4,4-dimethyl-6-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1,2,3,4-
tetrahydroquinoline 8b (70 mg, 0.157 mmol), 2-bromopyridine (12 µL,
0.12 mmol), Na2CO3 (32 mg, 0.30 mmol) and PdCl2(dppf) (6.2 mg,
0.08 mmol) in dioxane (0.80 mL) and water (0.21 mL) afforded, after
purification by flash chromatography on silica gel (cyclohexane/ethyl
acetate; 75/25), the title compound 9b as a yellow oil (45 mg, 94%).
HPLC purity: 99.9%, tR = 17.15 min. 1H NMR (300 MHz, DMSO-d6): δ
10.92 (s, 1H), 8.54 (d, J = 4.6 Hz, 1H), 7.93 (d, J = 2.1 Hz, 1H), 7.81 –
7.67 (m, 3H), 7.26 (dd, J = 5.4, 2.7 Hz, 2H), 7.18 – 7.11 (m, 1H), 7.05
(d, J = 2.3 Hz, 1H), 6.77 (d, J = 8.8 Hz, 1H), 6.72 (dd, J = 8.8, 2.4 Hz,
1H), 6.32 – 6.27 (m, 1H), 4.17 (t, J = 5.6 Hz, 2H), 3.76 (t, J = 5.5 Hz,
2H), 3.54 – 3.44 (m, 2H), 1.76 – 1.64 (m, 2H), 1.28 (s, 6H). 13C NMR
(75 MHz, DMSO-d6): δ 156.7, 152.3, 149.0, 144.5, 136.8, 131.1,
130.5, 128.0, 125.8, 125.3, 125.1, 123.9, 120.5, 118.4, 112.0, 111.5,
110.6, 102.8, 100.8, 65.1, 50.1, 46.0, 36.3, 31.7, 30.3 (2C). HRMS-
ESI (m/z): found 398.22189, calcd for C26H28N3O+ [M+H]+ 398.22269.
8c 1-(2-((1H-Indol-5-yl)oxy)ethyl)-4,4-dimethyl-7-(4,4,5,5-tetramethyl-
1,3,2-dioxaborolan-2-yl)-1,2,3,4-tetrahydroquinoline
Following method G, the reaction of 1-(2-((1H-indol-5-yl)oxy)ethyl)-7-
bromo-4,4-dimethyl-1,2,3,4-tetrahydroquinoline 6c (260 mg, 0.65
mmol), bis(pinacolato)diboron (198 mg, 0.78 mmol), KOAc (319 mg,
3.25 mmol) and PdCl2(dppf) (33.60 mg, 0.05 mmol) in dioxane (3.40
mL) afforded, after purification by flash chromatography on silica gel
(cyclohexane/ethyl acetate; 80/20), the title compound 8c as a white
solid (198 mg, 68%); mp 194 °C. 1H NMR (300 MHz, DMSO-d6): δ
10.91 (s, 1H), 7.28 – 7.22 (m, 2H), 7.14 (d, J = 7.6, 1H), 7.08 – 6.99
(m, 2H), 6.87 (d, J = 7.6 Hz, 1H), 6.74 (d, J = 8.6 Hz, 1H), 6.29 (br,
1H), 4.17 – 4.07 (m, 2H), 3.71 – 3.62 (m, 2H), 1.72 – 1.59 (m, 2H),
1.26 (s, 12H), 1.20 (s, 6H). 13C NMR (75 MHz, DMSO-d6): δ 152.3,
143.2, 134.1, 131.1, 128.0, 125.8, 125.2, 121.9, 119.4, 116.5, 111.9,
111.4, 102.8, 100.8, 83.3 (2C), 65.1, 50.3, 45.7, 36.3, 31.8, 30.3 (2C),
9c 1-(2-((1H-Indol-5-yl)oxy)ethyl)-4,4-dimethyl-7-(pyridin-2-yl)-1,2,3,4-
tetrahydroquinoline
Following method H, the reaction of 1-(2-((1H-indol-5-yl)oxy)ethyl)-
4,4-dimethyl-7-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1,2,3,4-
tetrahydroquinoline 8c (50 mg, 0.11 mmol), 2-bromopyridine (8.20 µL,
0.086 mmol), Na2CO3 (23 mg, 0.22 mmol) and PdCl2(dppf) (5 mg,
0.006 mmol) in dioxane (0.60 mL) and water (0.15 mL) afforded, after
purification by flash chromatography on silica gel (cyclohexane/ethyl
acetate; 75/25), the title compound 9c as a yellow oil (8 mg, 18%).
t
1
HPLC purity: 99.0 %, R = 17.63 min. H NMR (300 MHz, DMSO-d6):
δ 10.91 (br, 1H), 8.61 (d, J = 4.8 Hz, 1H), 7.81 – 7.73 (m, 2H), 7.38 (d,
J = 1.2 Hz, 1H), 7.33 – 7.18 (m, 5H), 7.07 (d, J = 2.4 Hz, 1H), 6.73 (dd,
J = 8.7, 2.4 Hz, 1H), 6.31 – 6.25 (m, 1H), 4.20 (t, J = 5.7 Hz, 2H), 3.79
(t, J = 5.7 Hz, 2H), 3.52 – 3.43 (m, 2H), 1.76 – 1.66 (m, 2H), 1.25 (s,
6H). 13C NMR (75 MHz, DMSO-d6): δ 156.9, 152.3, 149.3, 143.9,
137.1, 136.9, 131.7, 131.1, 128.0, 126.1, 125.8, 122.0, 120.0, 113.9,
112.0, 111.5, 108.6, 102.8, 100.8, 65.0, 50.2, 45.9, 36.4, 31.6, 30.4
(2C). HRMS-ESI (m/z): found 398.21985, cacld for C26H28N3O+
[M+H]+ 398.22269.
+
24.7 (4C). HRMS-ESI (m/z): found 447.27817, cacld for C27H36BN2O3
[M+H]+ 447.28135.
Method H: Suzuki coupling of 2-bromopyridine (9a-c).
A mixture of boronate ester 8 or 10 (1.3 equiv), 2-bromopyridine (1.0
equiv) and Na2CO3 (2.5 equiv) in dioxane and water was degassed
with argon. PdCl2(dppf) (0.07 equiv) was then added and the resulting
mixture was stirred 5 h at 100 °C. The mixture was cooled to rt,
filtered through a pad of dicalite and extracted with ethyl acetate.
Combined organic extracts were washed with brine and dried with
MgSO4. After evaporation under reduced pressure, the crude product
was purified by flash chromatography on silica gel.
10 1-(2-((1H-Indol-5-yl)oxy)ethyl)-5-(5,5-dimethyl-1,3,2-dioxaborinan-
2-yl)-4,4-dimethyl-1,2,3,4-tetrahydroquinoline
Following method G, the reaction of 1-(2-((1H-indol-5-yl)oxy)ethyl)-5-
bromo-4,4-dimethyl-1,2,3,4-tetrahydroquinoline 6a (400 mg, 1.00
mmol), bis(neopentyl glycolato)diboron (271 mg, 1.20 mmol), KOAc
(491 mg, 5.00 mmol) and PdCl2(dppf) (51 mg, 0.07 mmol) in dioxane
(8.60 mL) afforded, after purification by flash chromatography on silica
9a 1-(2-((1H-Indol-5-yl)oxy)ethyl)-4,4-dimethyl-5-(pyridin-2-yl)-1,2,3,4-
tetrahydroquinoline
10
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10.1002/cmdc.202000841
ChemMedChem
FULL PAPER
gel (cyclohexane/ethyl acetate; 80/20), the title compound 10 as a
light yellow solid (124 mg, 29%); mp 77 °C. HPLC purity: 96.6%, R =
12d’ 1-(2-((1-Benzyl-1H-indol-5-yl)oxy)ethyl)-4,4-dimethyl-8-(pyridin-
4-yl)-1,2,3,4-tetrahydroquinoline
t
24.31 min. 1H NMR (300 MHz, DMSO-d6): δ 10.89 (br, 1H), 7.28 –
7.23 (m, 2H), 7.03 (d, J = 2.4 Hz, 1H), 6.90 (dd, J = 8.4, 7.1 Hz, 1H),
6.70 (dd, J = 8.7, 2.4 Hz, 1H), 6.65 (dd, J = 8.4, 1.0 Hz, 1H), 6.47 (dd,
J = 7.1, 1.0 Hz, 1H), 6.31 – 6.27 (m, 1H), 4.12 (t, J = 5.7 Hz, 2H), 3.73
– 3.64 (m, 6H), 3.48 – 3.41 (m, 2H), 1.67 – 1.59 (m, 2H), 1.27 (s, 6H),
1.00 (s, 6H). 13C NMR (75 MHz, DMSO-d6): δ 152.4, 143.1, 137.5,
132.5, 131.1, 128.0, 125.8, 125.7, 119.5, 111.9, 111.4, 106.0, 102.7,
100.8, 71.5 (2C), 65.0, 50.5, 45.7, 38.0, 32.8, 31.3, 29.8 (2C), 21.5
Following method D, the reaction of 2-((1-benzyl-1H-indol-5-
yl)oxy)ethanol 4 (0.65 mg, 2.41 mmol), IBX (2.02 g, 2.28 mmol), 4,4-
dimethyl-8-(pyridin-4-yl)-1,2,3,4-tetrahydroquinoline 11d’ (1.21 g, 5.07
mmol), NaBH(OAc)3 (1.02 g, 4.82 mmol) in dry 1,2-DCE (10 mL)
afforded, after purification by flash chromatography on silica gel
(cyclohexane/ethyl acetate; 60/40), the title compound 12d’ as a
yellow oil (164 mg, 14%). 1H NMR (300 MHz, DMSO-d6): δ 8.58 –
8.54 (m, 2H), 7.47 – 7.44 (m, 2H), 7.43 (d, J = 3.0 Hz, 1H), 7.35 (dd, J
= 4.7, 4.7 Hz, 1H), 7.33 – 7.12 (m, 6H), 6.97 – 6.92 (m, 2H), 6.73 (d, J
= 2.4 Hz, 1H), 6.47 (dd, J = 8.9, 2.4 Hz, 1H), 6.35 (dd, J = 3.0, 0.7 Hz,
1H), 5.34 (s, 2H), 3.57 (t, J = 6.2 Hz, 2H), 3.29 – 3.19 (m, 2H), 2.95 (t,
J = 6.2 Hz, 2H), 1.75 – 1.64 (m, 2H), 1.29 (s, 6H). 13C NMR (75 MHz,
DMSO-d6): δ 152.1, 149.8, 149.7 (2C), 144.7, 138.4 (2C), 131.1,
130.6, 129.6, 128.8, 128.6, 128.5 (2C), 127.3, 126.9 (2C), 126.9,
123.6 (2C), 120.8, 111.5, 110.7, 103.2, 100.6, 65.9, 54.3, 49.2, 44.6,
33.9, 32.5, 31.0 (2C). HRMS-ESI (m/z): found 488.26873, cacld for
C33H34N3O+ [M+H]+ 488.26964.
+
(2C). HRMS-ESI (m/z): found 433.26455, cacld for C27H36BN2O3
[M+H]+ 433.26570.
11d 4,4-Dimethyl-8-(pyridin-3-yl)-1,2,3,4-tetrahydroquinoline
Following method F, the reaction of 8-bromo-4,4-dimethyl-1,2,3,4-
tetrahydroquinoline 3d (500 mg, 2.08 mmol), pyridin-3-ylboronic acid
(768 mg, 6.25 mmol), K2CO3 (864 mg, 6.25 mmol) and PdCl2(PPh3)2
(146 mg, 0.208 mmol) in DMF (30 mL) and water (6 mL) afforded,
after purification by flash chromatography on silica gel
(cyclohexane/ethyl acetate; 60/40), the title compound 11d as an
orange solid (389 mg, 78%); mp 88 °C. 1H NMR (300 MHz, DMSO-
d6): δ 8.55 – 8.50 (m, 2H), 7.78 (ddd, J = 7.8, 2.2, 1.8 Hz, 1H), 7.43
(ddd, J = 7.8, 4.8, 0.9 Hz, 1H), 7.19 (dd, J = 7.8, 1.4 Hz, 1H), 6.77 (dd,
J = 7.6, 1.4 Hz, 1H), 6.58 (dd, J = 7.8, 7.6 Hz, 1H), 5.01 (brs, 1H),
3.17 – 3.09 (m, 2H), 1.66 – 1.58 (m, 2H), 1.26 (s, 6H). 13C NMR (75
MHz, DMSO-d6): δ 149.8, 147.8, 141.3, 136.7, 135.6, 129.8, 128.0,
126.2, 123.7, 122.3, 115.6, 37.6, 36.4, 31.8, 31.1 (2C). HRMS-ESI
(m/z): found 239.15213, cacld for C16H19N2+ [M+H]+ 239.15428.
13d
1-(2-((1H-Indol-5-yl)oxy)ethyl)-4,4-dimethyl-8-(pyridin-3-yl)-
1,2,3,4-tetrahydroquinoline
Following method E, the reaction of 1-(2-((1-benzyl-1H-indol-5-
yl)oxy)ethyl)-4,4-dimethyl-8-(pyridin-3-yl)-1,2,3,4-tetrahydroquinoline
12d (100 mg; 0.21 mmol) and t-BuOK (323 mg , 2.87 mmol) in DMSO
(6.60 mL) afforded, after purification by flash chromatography on silica
gel (cyclohexane/ethyl acetate; 70/30), the title compound 13d as a
t
white solid (64 mg, 77%); mp 158 °C. HPLC purity: 97.3%, R = 18.54
11d’ 4,4-Dimethyl-8-(pyridin-4-yl)-1,2,3,4-tetrahydroquinoline
min. 1H NMR (300 MHz, DMSO-d6): δ 10.90 – 10.87 (m, 1H), 8.65 (dd,
J = 2.3, 0.7 Hz, 1H), 8.49 (dd, J = 4.8, 1.7 Hz, 1H), 7.87 (ddd, J = 7.8,
2.3, 1.7 Hz, 1H), 7.43 (ddd, J = 7.8, 4.8, 0.7 Hz, 1H), 7.37 (dd, J = 5.9,
3.6 Hz, 1H), 7.26 (dd, J = 2.8, 2.7 Hz, 1H), 7.19 (d, J = 8.7 Hz, 1H),
7.00 – 6.95 (m, 2H), 6.69 (d, J = 2.3 Hz, 1H), 6.47 (dd, J = 8.7, 2.3 Hz,
1H), 6.32 – 6.27 (m, 1H), 3.45 (t, J = 6.4 Hz, 2H), 3.25 – 3.19 (m, 2H),
2.93 (t, J = 6.4 Hz, 2H), 1.80 – 1.64 (m, 2H), 1.30 (s, 6H). 13C NMR
(75 MHz, DMSO-d6): δ 151.9, 149.5, 147.5, 145.3, 138.6, 137.5,
136.2, 131.1, 130.6, 129.1, 127.9, 126.7, 125.7, 123.5, 121.3, 111.8,
111.4, 102.6, 100.8, 66.0, 54.1, 44.5, 33.5, 32.5, 31.2 (2C). HRMS-
ESI (m/z): found 398.21939, cacld for C26H28N3O+ [M+H]+ 398.22269.
Following method F, the reaction of 8-bromo-4,4-dimethyl-1,2,3,4-
tetrahydroquinoline 3d (367 mg, 1.53 mmol), pyridin-4-ylboronic acid
(563 mg, 4.58 mmol), K2CO3 (633 mg, 4.58 mmol) and PdCl2(PPh3)2
(107 mg, 0.15 mmol) in DMF (22 mL) and water (4.50 mL) afforded,
after purification by flash chromatography on silica gel
(cyclohexane/ethyl acetate; 60/40), the title compound 11d’ as a
yellow solid (320 mg, 88%); mp 174 °C. 1H NMR (300 MHz, DMSO-
d6): δ 8.60 – 8.56 (m, 2H), 7.42 – 7.38 (m, 2H), 7.20 (dd, J = 7.6, 1.6
Hz, 1H), 6.80 (dd, J = 7.6, 1.6 Hz, 1H), 6.58 (dd, J = 7.6, 7.6 Hz, 1H),
5.13 (s, 1H), 3.19 – 3.10 (m, 2H), 1.67 – 1.59 (m, 2H), 1.26 (s, 6H).
13C NMR (75 MHz, DMSO-d6): δ 149.9 (2C), 147.6, 140.8, 130.0,
127.6, 126.6, 124.1 (2C), 123.0, 115.6, 37.6, 36.3, 31.8, 31.0 (2C).
13d’
1-(2-((1H-Indol-5-yl)oxy)ethyl)-4,4-dimethyl-8-(pyridin-4-yl)-
1,2,3,4-tetrahydroquinoline
+
HRMS-ESI (m/z): found 239.15334, cacld for C16H19N2 [M+H]+
239.15428.
Following method E, the reaction of 1-(2-((1-benzyl-1H-indol-5-
yl)oxy)ethyl)-4,4-dimethyl-8-(pyridin-4-yl)-1,2,3,4-tetrahydroquinoline
12d’ (150 mg; 0.31 mmol) and t-BuOK (484 mg , 4.31 mmol) in
DMSO (9.60 mL) afforded, after purification by flash chromatography
on silica gel (cyclohexane/ethyl acetate; 70/30), the title compound
12d 1-(2-((1-Benzyl-1H-indol-5-yl)oxy)ethyl)-4,4-dimethyl-8-(pyridin-3-
yl)-1,2,3,4-tetrahydroquinoline
t
13d’ as a yellow oil (94 mg, 77%). HPLC purity: 99.6%, R = 18.20
Following method D, the reaction of 2-((1-benzyl-1H-indol-5-
yl)oxy)ethanol 4 (0.37 g, 1.37 mmol), IBX (1.15 g, 4.11 mmol), 4,4-
dimethyl-8-(pyridin-3-yl)-1,2,3,4-tetrahydroquinoline 11d (0.69 g, 2.89
mmol), NaBH(OAc)3 (0.58 g, 2.74 mmol) in dry 1,2-DCE (6.00 mL)
afforded, after purification by flash chromatography on silica gel
(cyclohexane/ethyl acetate; 70/30), the title compound 12d as a light
min. 1H NMR (300 MHz, DMSO-d6): δ 10.88 (brs, 1H), 8.60 – 8.56 (m,
2H), 7.50 – 7.46 (m, 2H), 7.36 (dd, J = 5.9, 3.5 Hz, 1H), 7.26 (dd, J =
2.8, 2.7 Hz, 1H), 7.19 (d, J = 8.8 Hz, 1H), 6.99 – 6.92 (m, 2H), 6.72 (d,
J = 2.4 Hz, 1H), 6.47 (dd, J = 8.8, 2.4 Hz, 1H), 6.31 – 6.27 (m, 1H),
3.57 (t, J = 6.2 Hz, 2H), 3.30 – 3.21 (m, 2H), 2.96 (t, J = 6.2 Hz, 2H),
1.75 – 1.65 (m, 2H), 1.30 (s, 6H). 13C NMR (75 MHz, DMSO-d6): δ
151.9, 149.8, 149.7 (2C), 144.8, 138.4, 131.1, 130.7, 128.9, 127.9,
126.9, 125.8, 123.7 (2C), 120.9, 111.8, 111.4, 102.7, 100.8, 65.9,
54.4, 44.6, 33.9, 32.5, 31.1 (2C). HRMS-ESI (m/z): found 398.22138,
cacld for C26H28N3O+ [M+H]+ 398.22269.
1
yellow oil (100 mg, 15%). H NMR (300 MHz, DMSO-d6): δ 8.68 (brs,
1H), 8.50 (brd, J = 4.0 Hz, 1H), 7.90 (ddd, J = 7.9, 1.9, 1.8 Hz, 1H),
7.46 – 7.42 (m, 2H), 7.36 (dd, J = 6.1, 3.3 Hz, 1H), 7.33 – 7.21 (m,
4H), 7.19 – 7.13 (m, 2H), 7.00 – 6.94 (m, 2H), 6.72 (d, J = 2.4 Hz, 1H),
6.49 (dd, J = 8.8, 2.4 Hz, 1H), 6.36 (d, J = 3.1 Hz, 1H), 5.34 (s, 2H),
3.46 (t, J = 6.3 Hz, 2H), 3.26 – 3.16 (m, 2H), 2.92 (t, J = 6.3 Hz, 2H),
1.73 – 1.63 (m, 2H), 1.30 (s, 6H). 13C NMR (75 MHz, DMSO-d6): δ
152.2, 149.0, 145.0, 145.2, 138.6, 138.4, 137.7, 136.8, 131.0, 130.4,
129.6, 129.2, 128.6, 128.5 (2C), 127.3, 126.9 (2C), 126.8, 123.8,
121.4, 111.6, 110.7, 103.2, 100.6, 66.0, 54.0, 49.2, 44.4, 33.5, 32.5,
31.2 (2C). HRMS-ESI (m/z): found 488.26583, cacld for C33H34N3O+
[M+H]+ 488.26964.
BIOLOGY
Cell cultures and reagents
11
This article is protected by copyright. All rights reserved.

10.1002/cmdc.202000841
ChemMedChem
FULL PAPER
K562, KU812, KCL-22, IM-sensitive (K562S), IM-resistant (K562R)
BCR-ABL+ cells and MV4-11 cells were obtained from the Deutshe
Sammlung von Mikroorganismens und Zellkulturen (DSMZ). MOLM-
13 and HS-27a were obtained from the American Type Culture
Collection (ATCC). All cell lines were obtained according to the
supplier’s recommendations. Cells were cultured in RPMI 1640 (Gibco
– Thermofisher, Waltham, MA, USA) , with 10% fetal bovine serum
(Thermoscientific – Sigma Aldrich St Louis, MO, USA), 2 mM
glutamine (Gibco – Thermofisher, Waltham, MA, USA), 100 U/mL
RNA samples were reverse-transcribed using SuperScript®VILO
cDNA synthesis kit (Invitrogen, Carlsbad, USA) as recommended by
the supplier. The resulting cDNAs were used for quantitative real-time
PCR
(qRT-PCR).
PCR
primers
(PIM1:
rev
for
5’-
5’-
5’-
5’-
5’-
5’-
5’-
5’-
the
TTTCGAGCATGACGAAGAGA-3’,
GGGCCAAGCACCATCTAAT-3’;
CISH:
for
for
AGCCAAGACCTTCTCCTACCTT-3’,
TGGCATCTTCTGCAGGTGT-3’;
TCCCTATAACATGTACCCACA-3’,
ATGGTCTCATCCAGGTCGAA-3’;
TGAAGGCCACCATCATCAG-3’,
TGTTCAAGATCTCGCCACTG-3’)
rev
STAT5A:
rev
STAT5B:
rev
designed
penicillin and 100 µg/mL streptomycin (Gibco
–
Thermofisher,
for
with
Waltham, MA, USA) at 37 °C, 5% CO2. Resistant cells were cultured
with 1 µM IM. IM was purchased from Selleckchem (Houston, USA).
Bone marrow (BM) MSC, obtained from donors undergoing
orthopedic surgery were isolated and cultured as described.[47]
Informed consent was obtained before BM samples were taken. MSC
were cultured in αMEM (Gibco – Thermofisher, Waltham, MA, USA) ,
with 10% fetal bovine serum (Thermoscientific – Sigma Aldrich St
Louis, MO, USA), 2 mM glutamine (Gibco – Thermofisher, Waltham,
MA, USA), 100 U/mL penicillin, 100 µg/mL streptomycin (Gibco –
were
ProbeFinder software (Roche Applied Sciences, Basel, Switzerland)
and used to amplify the RT-generated cDNAs. qRT-PCR analyses
were performed on the Light Cycler 480 thermocycler II (Roche).
GAPDH (glyceraldehyde-3-phosphate dehydrogenase), ACTB (actin
beta) and RPL13A (ribosomal protein L13A) were used as reference
genes for normalization of qRT-PCR experiments. Each reaction
condition was performed in triplicate. Relative gene expression was
analyzed using the 2-ΔΔCt method.[48]
Thermofisher, Waltham, MA, USA), 0.01% fungizone (Gibco
–
Thermofisher, Waltham, MA, USA) and 0.005% fibroblast growth
factor 2 (Gibco – Thermofisher, Waltham, MA, USA) at 37 °C, 5%
CO2.
Acknowledgements
Cell proliferation assays
This work was supported by the Region Centre - Val de Loire,
the Fondation pour la Recherche Médicale (FRM, grant
number: DCM20181039564), the Laboratory of excellence
Labex SYNORG, ANR-11-LABX-0029 and La Ligue Contre le
Cancer. We thank Emmanuel Pecnard for technical
assistance
Cell viability and proliferation were studied using
a MTT cell
proliferation assay (Sigma-Aldrich, MO, USA). Briefly, 2x104 leukemic
cells were cultured in 100 µL of RPMI medium in 96 well plates and
treated with 10 µM of each drug for 24, 48 and 72 h. To determine the
concentration-effect of the molecules, cells were treated with
concentrations ranging from 100 nM to 100 µM for 48 h. To determine
drugs toxicity on stromal cells, 1x104 HS-27a or 3.5x103 were cultured
Keywords: Inhibitors - Medicinal Chemistry - Myeloid
in 96 well plates, incubated for 24
h and then treated with
Leukemias - STAT5 - Suzuki coupling
concentrations ranging from 100 nM to 100 µM of drugs for 48 h.
Cells were then incubated with 10 µL of MTT working solution (5 g/L
of methylthiazolyldiphenyl-tetrazolium bromide) during 4 h. Cells were
lysed overnight at 37 °C with 100 μL of SDS 10%, HCl 0.003%.
Supplementary Data
Optical density (OD) at 570 nm was then measured using
a
spectrophotometer CLARIOstar® (BMG Labtech, Offenburg,
Germany). Living cells were also enumerated using the trypan blue
dye exclusion method.
Detailed synthesis strategic pathways starting from 2-
aminophenylboronic acid and 2-chloroaniline to reach 2-
pyridinyl analog in position 8 on THQ ring, NMR spectra and
biological evaluation of compounds 6c, 10, 13d and 13d’ on
other AML cell lines, KG1a and MOLM-13.
Apoptosis analysis
KU812 and MV4-11 cells were treated for 48 h with 3 µM of each
drugs and then washed with PBS, then stained (106 cells) in buffer
containing FITC-annexin
V and 7-amino-actinomycin D (7-AAD)
Abbreviations used
(Beckmann Coulter, Fullerton, USA) for 15 min at 4 °C and analyzed
by flow cytometry (Becton Dickinson Accuri™ C6 flow cytometer). The
FlowJo® software was used to analyze data.
AKT, Protein Kinase B; AML, Acute Myeloid Leukemia; BCR-
ABL,
Breakpoint
Cluster
Region-Abelson;
Bneop,
neopentylglycolato borane; Bpin, pinacol borane CML,
Chronic Myeloid Leukemia; DCE, Dichloroethane; DMF,
Dimethylformamide; DMSO, Dimethyl sulfoxide; DNA,
Deoxyribonucleic acid; FLT3-ITD, FMS like tyrosine kinase 3 -
Internal Tandem Duplication; IBX, 2-iodoxybenzoic acid; IM,
Imatinib Mesylate; JAK2, Janus Kinase 2; PIM1, Proviral
Integration site for Moloney leukemia virus 1; PPARγ,
Peroxisome Proliferator – Activated Receptor gamma; RNAi,
interference ribonucleic acid; ROS, Reactive Oxygen
Species; SAR, Structure-Activity Relationship; SMA-ZnPP,
Styrene-Maleic Acid-copolymer conjugated ZnPP; STAT5,
Signal transducer and activator of transcription 5; TFA,
Trifluoroacetic Acid; THQ, Tetrahydroquinoline; TKI, Tyrosine
Kinase Inhibitor.
Western blot
Cells were suspended in NP40 buffer (Bio-Rad, NY, USA), separated
on SDS/PAGE and blotted onto nitrocellulose membrane. Blots were
incubated with the following antibodies (Abs): P-Y694/699-STAT5,
Actin (Cell signaling Technology, Danvers, USA), STAT5 (BD
Transduction Laboratories, Franklin Lakes, USA), STAT5A and
STAT5B (Zymed/ThermoFisher Scientific, Waltham, MA, USA).
Membranes were developed with the ECL chemiluminescence
detection system (GE Healthcare, Little Chalfont, UK) using specific
peroxidase (HRP) conjugated to rabbit or mouse IgG antibodies (Cell
signaling Technology, Danvers, USA).
qRT-PCR analysis
12
This article is protected by copyright. All rights reserved.

10.1002/cmdc.202000841
ChemMedChem
FULL PAPER
References
Hédou, M.-C. Viaud-Massuard, G. Prié, F. Gouilleux, Cancers
(Basel). 2020, 12, 240–287.
[1]
[2]
K. D. Bunting, Front. Biosci. 2007, 12, 2807–2820.
[24]
[25]
C. Sillaber, F. Gesbert, D. A. Frank, M. Sattler, J. D. Griffin,
Blood 2000, 95, 2118–2125.
C. L. Shaui, K.ꢀ; Halpern, J.ꢀ; ten Hoeve, J.ꢀ; Rao, X.ꢀ; Sawyers,
Oncogene 1996, 13, 247–254.
A. Hoelbl, C. Schuster, B. Kovacic, B. Zhu, M. Wickre, M. A.
Hoelzl, S. Fajmann, F. Grebien, W. Warsch, G. Stengl, L.
Hennighausen, V. Poli, H. Beug, R. Moriggl, V. Sexl, EMBO Mol.
Med. 2010, 2, 98–110.
[3]
[4]
[5]
M. Mizuki, R. Fenski, H. Halfter, I. Matsumura, R. Schmidt, C.
Muller, W. Gruning, K. Kratz-Albers, S. Serve, C. Steur, T.
Buchner, J. Kienast, Y. Kanakura, W. E. Berdel, H. Serve, Blood
2000, 96, 3907–3914.
[26]
D. Ye, N. Wolff, L. Li, S. Zhang, R. L. Ilaria, Blood 2006, 107,
4917–4925.
C. James, V. Ugo, J. P. Le Couédic, J. Staerk, F. Delhommeau,
C. Lacout, L. Garçon, H. Raslova, R. Berger, A. Bennaceur-
Griscelli, J. L. Villeval, S. N. Constantinescu, N. Casadevall, W.
Vainchenker, Nature 2005, 434, 1144–1148.
[27]
[28]
D. Fabbro, Nat. Chem. Biol. 2012, 8, 228–229.
A. Brady, S. Gibson, L. Rybicki, E. Hsi, Y. Saunthararajah, M. A.
Sekeres, R. Tiu, E. Copelan, M. Kalaycio, R. Sobecks, J. Bates,
A. S. Advani, Eur. J. Haematol. 2012, 89, 288–293.
J. Müller, B. Sperl, W. Reindll, A. Kiessling, T. Berg,
ChemBioChem 2008, 9, 723–727.
M. Kontro, H. Kuusanmäki, S. Eldfors, T. Burmeister, E. I.
Andersson, Bruserud, T. H. Brümmendorf, H. Edgren, B. T.
Gjertsen, M. Itälä-Remes, S. Lagström, O. Lohi, T. Lundán, J. M.
L. Martí, M. M. Majumder, A. Parsons, T. Pemovska, H. Rajala,
K. Vettenranta, O. Kallioniemi, S. Mustjoki, K. Porkka, C. A.
Heckman, Leukemia 2014, 28, 1738–1742.
[29]
[30]
[31]
N. Elumalai, A. Berg, K. Natarajan, A. Scharow, T. Berg, Angew.
Chemie Int. Ed. 2015, 54, 4758–4763.
B. D. G. Page, H. Khoury, R. C. Laister, S. Fletcher, M. Vellozo,
A. Manzoli, P. Yue, J. Turkson, M. D. Minden, P. T. Gunning, J.
Med. Chem. 2012, 55, 1047–1055.
[6]
B. J. Druker, S. Tamura, E. Buchdunger, S. Ohno, G. M. Segal,
S. Fanning, J. Zimmermann, N. B. Lydon, Nat. Med. 1996, 2,
561–566.
[32]
[33]
A. A. Cumaraswamy, A. M. Lewis, M. Geletu, A. Todic, D. B.
Diaz, X. R. Cheng, C. E. Brown, R. C. Laister, D. Muench, K.
Kerman, H. L. Grimes, M. D. Minden, P. T. Gunning, ACS Med.
Chem. Lett. 2014, 5, 1202–1206.
[7]
[8]
N. P. Shah, C. Tran, F. Y. Lee, P. Chen, D. Norris, C. L.
Sawyers, Science. 2004, 305, 399–401.
E. Weisberg, P. W. Manley, W. Breitenstein, J. Brüggen, S. W.
Cowan-Jacob, A. Ray, B. Huntly, D. Fabbro, G. Fendrich, E.
Hall-Meyers, A. L. Kung, J. Mestan, G. Q. Daley, L. Callahan, L.
Catley, C. Cavazza, A. Mohammed, D. Neuberg, R. D. Wright, D.
G. Gilliland, J. D. Griffin, Cancer Cell 2005, 7, 129–141.
J. M. Golas, K. Arndt, C. Etienne, J. Lucas, D. Nardin, J.
Gibbons, P. Frost, F. Ye, D. H. Boschelli, F. Boschelli, Cancer
Res. 2003, 63, 375–381.
B. Wingelhofer, B. Maurer, E. C. Heyes, A. A. Cumaraswamy, A.
Berger-Becvar, E. D. De Araujo, A. Orlova, P. Freund, F. Ruge,
J. Park, G. Tin, S. Ahmar, C. H. Lardeau, I. Sadovnik, D. Bajusz,
G. M. Keseru, F. Grebien, S. Kubicek, P. Valent, P. T. Gunning,
R. Moriggl, Leukemia 2018, 32, 1135–1146.
[9]
[34]
Z. Liao, L. Gu, J. Vergalli, S. A. Mariani, M. De Dominici, R. K.
Lokareddy, A. Dagvadorj, P. Purushottamachar, P. A. McCue, E.
Trabulsi, C. D. Lallas, S. Gupta, E. Ellsworth, S. Blackmon, A.
Ertel, P. Fortina, B. Leiby, G. Xia, H. Rui, D. T. Hoang, L. G.
Gomella, G. Cingolani, V. Njar, N. Pattabiraman, B. Calabretta,
M. T. Nevalainen, Mol. Cancer Ther. 2015, 14, 1777–1793.
E. A. Nelson, S. R. Walker, E. Weisberg, M. Bar-Natan, R.
Barrett, L. B. Gashin, S. Terrell, J. L. Klitgaard, L. Santo, M. R.
Addorio, B. L. Ebert, J. D. Griffin, D. A. Frank, Blood 2011, 117,
3421–3429.
[10]
C. L. Shamroe, J. M. Comeau, Ann. Pharmacother. 2013, 47,
1540–1546.
[11]
[12]
T. Tauchi, K. Ohyashiki, Leuk. Res. 2004, 28, 6–7.
R. Nimmanapalli, K. Bhalla, Curr. Opin. Oncol. 2002, 14, 616–
620.
[35]
[36]
[37]
[13]
S. W. Cowan-Jacob, V. Guez, G. Fendrich, J. D. Griffin, D.
Fabbro, P. Furet, J. Liebetanz, J. Mestan, P. W. Manley, Mini
Rev. Med. Chem. 2004, 4, 285–299.
[14]
[15]
A. Hochhaus, Semin. Hematol. 2003, 40, 69–79.
S.-Y. . Kang, K.-H.; Kim, S.-H.ꢀ; Choi, S.-Y.; Yoo, H.-L.; Lee, M.-
Y.ꢀ; Song, H.-Y.; Kee, K.-M.; Suh, J.-H.; Yang, D.-W. Jang, E.-J.;
Lee, S.-E.; Kim, Leuk. Res. 2019, 76, 87–93.
E. A. Nelson, S. R. Walker, M. Xiang, E. Weisberg, M. Bar-
Natan, R. Barrett, S. Liu, S. Kharbanda, A. L. Christie, M.
Nicolais, J. D. Griffin, R. M. Stone, A. L. Kung, D. A. Frank,
Genes Cancer 2012, 3, 503–511.
[16]
D. C. Kottaridis, P. D.; Gale, R. E.; Frew, M. E.; Harrison, G.;
Langabeer, S. E.; Belton, A. A.; Walker, H.; Wheatley, K.; Bowen,
D. T.; Burnett, A. K.; Goldstone, A. H.; Linch, Blood 2001, 98,
1752–1760.
R. Rondanin, D. Simoni, M. Maccesi, R. Romagnoli, S.
Grimaudo, R. M. Pipitone, M. Meli, A. Cascio, M. Tolomeo,
ChemMedChem 2017, 12, 1183–1190.
[38]
[39]
J. M. Shipley, D. J. Waxman, Toxicol. Appl. Pharmacol. 2004,
199, 275–284.
[17]
[18]
[19]
[20]
R. M. Stone, Semin. Hematol. 2002, 39, 4–10.
J. L. Shipley, J. N. Butera, Exp. Hematol. 2009, 37, 649–658.
M. Andreeff, Expert Rev. Anticancer Ther. 2015, 15, 1–15.
C. M. Thomas, P. Campbell, J. Oncol. Pharm. Pract. 2018, 0, 1–
9.
S. Prost, M. Le Dantec, S. Augé, R. Le Grand, S. Derdouch, G.
Auregan, N. Déglon, F. Relouzat, A. Aubertin, B. Maillere, I.
Dusanter-fourt, M. Kirszenbaum, J. Clin. Invest. 2008, 118,
1765–1775.
[21]
[22]
[23]
N. Daver, R. F. Schlenk, N. H. Russell, M. J. Levis, Leukemia
2019, 33, 299–312.
[40]
S. Prost, F. Relouzat, M. Spentchian, Y. Ouzegdouh, J. Saliba,
G. Massonnet, J. P. Beressi, E. Verhoeyen, V. Raggueneau, B.
Maneglier, S. Castaigne, C. Chomienne, S. Chrétien, P.
Rousselot, P. Leboulch, Nature 2015, 525, 380–383.
A. Berger, V. Sexl, P. Valent, R. Moriggl, Oncotarget 2014, 5,
9564–9576.
M. Brachet-Botineau, M. Polomski, H. A. Neubauer, L. Juen, D.
13
This article is protected by copyright. All rights reserved.

10.1002/cmdc.202000841
ChemMedChem
FULL PAPER
[41]
[42]
L. Juen, M. Brachet-Botineau, C. Parmenon, J. Bourgeais, O.
Hérault, F. Gouilleux, M.-C. Viaud-Massuard, G. Prié, J. Med.
Chem. 2017, 60, 6119–6136.
M. Brachet-Botineau, M. Deynoux, N. Vallet, M. Polomski, L.
Juen, O. Hérault, F. Mazurier, M.-C. Viaud-Massuard, G. Prié, F.
Gouilleux, Cancers. 2019, 11, 2043–2059.
[43]
[44]
W. Zhu, Q. Hu, N. Hanke, C. J. Van Koppen, R. W. Hartmann, J.
Med. Chem. 2014, 57, 7811–7817.
M. Boehm, E. Martinborough, M. Moorjani, J. Tamiya, L. Huang,
A. Yeager, E. Brahmachary, T. Fowler, A. Novak, P. Meghani, M.
Knaggs, Novel GLP-1 Receptor Modulators, 2014, WO
2014201172.
[45]
H. T. T. Pham, B. Maurer, M. Prchal-Murphy, R. Grausenburger,
E. Grundschober, T. Javaheri, H. Nivarthi, A. Boersma, T. Kolbe,
M. Elabd, F. Halbritter, J. Pencik, Z. Kazemi, F. Grebien, M.
Hengstschläger, L. Kenner, S. Kubicek, M. Farlik, C. Bock, P.
Valent, M. Müller, T. Rülicke, V. Sexl, R. Moriggl, J. Clin. Invest.
2018, 128, 387–401.
[46]
T. Khanam, S. Sandmann, J. Seggewiss, C. M. Ruether, M.
Zimmermann, A. B. Norvil, C. Bartenhagen, G. Randau, S.
Mueller, H. Herbrüggen, P. Hoffmann, S. Herms, L. Wei, M.
Wöste, C. Wünsch, H. Gowher, I. Oschlies, W. Klapper, W.
Woessmann, M. Dugas, B. Burkhardt, Blood 2020, DOI
10.1182/blood.2020005381.
[47]
[48]
A. L. Ponte, T. Ribeiro-Fleury, V. Chabot, F. Gouilleux, A.
Langonné, O. Hérault, P. Charbord, J. Domenech, Stem Cells
Dev. 2012, 21, 3162–3172.
K. J. Livak, T. D. Schmittgen, Methods 2001, 25, 402–408.
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10.1002/cmdc.202000841
ChemMedChem
FULL PAPER
Entry for the Table of Contents
STAT5 inhibitors in myeloid leukemias: To improve the
antileukemic activity of a first lead compound called 17f, new
derivatives were synthesized by pharmacomodulation. Two
compounds, 7a and 7a’, showed similar or higher
antileukemic effects in various acute and chronic myeloid
leukemias cell lines. Both compounds were found to be more
effective than 17f to inhibit STAT5 activity/expression and to
suppress the chemoresistance of CML cells.
Keywords: Inhibitors - Medicinal Chemistry - Myeloid
Leukemias - STAT5 - Suzuki coupling
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