Discovery of AZD3147: A potent, selective dual inhibitor of mTORC1 and mTORC2

Article abstract of DOI:10.1021/jm501778s

High throughput screening followed by a lead generation campaign uncovered a novel series of urea containing morpholinopyrimidine compounds which act as potent and selective dual inhibitors of mTORC1 and mTORC2. We describe the continued compound optimization campaign for this series, in particular focused on identifying compounds with improved cellular potency, improved aqueous solubility, and good stability in human hepatocyte incubations. Knowledge from empirical SAR investigations was combined with an understanding of the molecular interactions in the crystal lattice to improve both cellular potency and solubility, and the composite parameters of LLE and pIC₅₀-pSolubility were used to assess compound quality and progress. Predictive models were employed to efficiently mine the attractive chemical space identified resulting in the discovery of 42 (AZD3147), an extremely potent and selective dual inhibitor of mTORC1 and mTORC2 with physicochemical and pharmacokinetic properties suitable for development as a potential clinical candidate.

Full text of DOI:10.1021/jm501778s

Article
pubs.acs.org/jmc
Discovery of AZD3147: A Potent, Selective Dual Inhibitor of mTORC1
and mTORC2
Kurt G. Pike,* Jeff Morris, Linette Ruston, Sarah L. Pass, Ryan Greenwood, Emma J. Williams,
Julie Demeritt, Janet D. Culshaw, Kristy Gill, Martin Pass, M. Raymond V. Finlay, Catherine J. Good,
Craig A. Roberts, Gordon S. Currie, Kevin Blades, Jonathan M. Eden, and Stuart E. Pearson
Oncology Innovative Medicines, AstraZeneca, Alderley Park, Macclesfield, Cheshire SK10 4TG, U.K.
ABSTRACT: High throughput screening followed by a lead generation campaign
uncovered a novel series of urea containing morpholinopyrimidine compounds which
act as potent and selective dual inhibitors of mTORC1 and mTORC2. We describe the
continued compound optimization campaign for this series, in particular focused on
identifying compounds with improved cellular potency, improved aqueous solubility,
and good stability in human hepatocyte incubations. Knowledge from empirical SAR
investigations was combined with an understanding of the molecular interactions in the
crystal lattice to improve both cellular potency and solubility, and the composite
parameters of LLE and pIC50−pSolubility were used to assess compound quality and
progress. Predictive models were employed to efficiently mine the attractive chemical
space identified resulting in the discovery of 42 (AZD3147), an extremely potent and selective dual inhibitor of mTORC1 and
mTORC2 with physicochemical and pharmacokinetic properties suitable for development as a potential clinical candidate.
treated with rapamycin.12 This suggests that by inhibiting
mTORC2 in addition to mTORC1, and thus having a direct
inhibitory effect on Akt signialling, a compound may offer
greater clinical benefit compared with rapalogues. In addition,
dual mTORC1 and mTORC2 inhibitors may exhibit a
broader spectrum of clinical activity.
INTRODUCTION
■
The mammalian target of rapamycin (mTOR) is a key target
in the development of antitumor therapies.1 Activated by
growth factor/mitogenic stimulation activation of the
phosphatidylinositol 3-kinase (PI3K)/Akt signaling pathway,
mTOR is a central regulator of cell growth and proliferation.
This PI3K-Akt-mTOR pathway is one of the most frequently
dysregulated pathways in cancer.2 mTOR, a serine/theronine
kinase of approximately 289 kDa in size, is a member of the
evolutionary conserved eukaryotic PI3K-related kinase (PIKK)
family of atypical protein kinases which also include the
protein kinases DNA-PK (DNA dependent protein kinase),
ATM (ataxia-telangiectasia mutated) and ATR (ataxia-
telangiectasia and Rad3 related).3−5
The known mTOR inhibitor rapamycin and its analogues
(RAD001, CCI-779, AP23573) bind to the FKBP12/
rapamycin complex binding domain (FRB), resulting in
suppression of signaling to the downstream targets p70S6K
and 4E-BP1.6,7 The potent but nonspecific inhibitors of PI3K,
LY294002 and wortmannin, have also been shown to inhibit
the kinase function of mTOR; however, in this case the
catalytic domain of the protein is targeted.8
Recently it has been shown that mTOR can exist in an
alternative, rapamycin insensitive complex that signals to Akt.9
The existence of both a rapamycin sensitive complex
(mTORC1) and a rapamycin insensitive complex
(mTORC2) may provide an explanation for the differences
observed in the earlier work of Brunn et al.8 and Edinger et
al.10 Rapamycin and its analogues have been shown to activate
AKT signaling as a consequence of inhibition of the negative
feedback loop downstream of mTORC1,11 which is associated
with a shorter time to progression in glioblastoma patients
A number of compounds are known to inhibit mTOR and
other members of the PIKK family and have been reviewed
extensively elsewhere.13 Early reported ATP-competitive
inhibitors of mTOR were found to inhibit the activity of a
number of different PIKK enzymes although were generally
selective when tested more broadly against other kinase
targets, such as LY294002 which is a moderate inhibitor of
PI3K, mTOR, and DNA-PK. Following significant investment
from a number of groups, more potent dual PI3K/mTOR
inhibitors have been discovered, such as NVP-BEZ235 and PI-
103. Compounds with a high degree of selectivity for mTOR
over the PI3K isoforms and the other PIKK enzymes have
also subsequently been reported, such as AZD8055, PP242,
AZD2014, and WYE-354 (Figure 1). More recently
compounds have been disclosed that show selectivity for
other members of the PIKK family such as Ku-0060648, a
potent and selective inhibitor of DNAPK. While NVP-
BEZ235 and PP242 contain classical kinase hinge binding
motifs, compounds such as LY294002, PI-103, AZD8055,
AZD2014, WYE-354, and Ku-0060648 are believed to bind to
the kinase hinge region through the morpholine oxygen in a
fashion analogous to that observed for LY294002 in PI3Kγ.14
Received: November 16, 2014
© XXXX American Chemical Society
A
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Figure 1. Structures of reported ATP-competitive PIKK inhibitors.
A recent crystal structure of PI-103 bound into the mTOR
protein further supports this binding mode.15
We have previously reported the identification of
compound 1 following a screen of the AstraZeneca compound
collection against a recombinant mTOR enzyme assay (Figure
2a).16 Although of only modest potency against mTOR
(pIC50 = 5.85), compound 1 was considered of particular
interest because of its significantly lower affinity for PI3Kα
(pIC50 = 4.76).17 Optimization of this novel scaffold resulted
in the identification of 2 showing increased mTOR affinity
(pIC50 = 6.54) and selectivity over PI3Kα (pIC50 = 4.58).
The importance of the indole hydrogen bond donor in 2 was
established through structure−activity relationship (SAR)
studies and rationalized using a homology model of the
mTOR ATP binding site built upon the ATP binding site of
PI3Kγ.14,18 The indole is believed to form a productive
interaction with a glutamic acid residue present in mTOR but
not PI3K (Figure 2b).19 Comparison of the homology model
with the recently reported crystal structure of mTOR15 reveals
no significant differences in protein structure and further
validates the use of the homology model to help guide design.
Subsequent optimization looked to maintain this key
interaction and resulted in the discovery of the urea
containing compound 3 in which mTOR affinity has been
further increased (pIC50 = 7.55) while selectivity over PI3Kα
has been maintained (pIC50 = 6.25). Compound 3 is a potent
inhibitor of substrates of both mTORC1 (phosphorylation of
Ser 235/236 on S6) and mTORC2 (phosphorylation of Ser
473 on Akt) in MDA-MB-468 cells (pIC50 of 6.92 and 7.06,
respectively).20 However, the low aqueous solubility of 3 (<1
μM) was felt to preclude this compound from further
development.21a,b Herein we describe our continuing efforts to
identify potent and selective dual inhibitors of mTORC1 and
mTORC2 which possess physicochemical properties suitable
for development as a clinical candidate.
Figure 2. (a) Identification of urea containing selective dual
mTORC1/2 inhibitors. (b) Proposed binding mode of compound
2 in mTOR homology model. D835 from PI3Kγ structure (1e7v) is
highlighted in red. Waters observed in PI3Kγ crystal structures that
could influence binding interactions are displayed as red spheres.
CHEMISTRY
■
Compounds 5, 6, 8−17, and 23 were synthesized according
the route outlined in Scheme 1.22,23 Reaction of methyl
dichloropyrimidinebenzoate with 3(S)-methylmorpholine and
triethylamine at room temperature resulted in the selective
installation of the amine in the 4-position to give intermediate
A in good yield. Reduction of the ester directly to the alcohol
using lithium borohydride gave intermediate B also in good
yield. Conversion of the alcohol to the iodide, intermediate C,
B
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a
Scheme 1. General Sysnthesis of 5, 6, 8−17, and 23
a
Reagents: (a) 3(S)-methylmorpholine, Et3N, DCM, rt. (b) LiBH4, THF, 0 °C. (c) (1) MsCl, Et3N, DCM, 0 °C; (2) NaI, DCM, 50 °C. (d)
Methanesufinic acid sodium salt, MeCN, 85 °C. (e) (4-Aminophenyl)boronic acid pinacol ester, PdCl2(PPh3)2, Na2CO3, DMF/DME/water/EtOH,
90 °C. (f) Appropriate isocyanate, 1,4-dioxane, 70 °C. (g) (1) Phosgene, rt; (2) appropriate amine, rt. (h) (1) Phenyl chloroformate, Na2CO3, 1,4-
dioxane, rt; (2) appropriate amine, Et3N, NMP or DMF, 50−70 °C. (j) (1) Di(imidazol-1-yl)methanethione, DCM, rt; (2) cyclopropylamine, Et3N,
rt.
a
Scheme 2. Synthesis of Compounds 4, 7, 24, and 25
a
Reagents: (a) ethyl isocyanate, 1,4-dioxane, 70 °C. (b) For the preparation of G: (4-amino-3-methoxyphenyl)boronic acid, PdCl2(PPh3)2, Na2CO3,
DMF/DME/water/EtOH, 90 °C. For the preparation of H: (4-amino-3-methylphenyl)boronic acid pinacol ester (prepared in situ by reaction of 4-
bromo-2-methylaniline with bis(pinacolato)diboron, PdCl2(PPh3)2, and Na2CO3 in 1,4-dioxane at 90 °C), PdCl2(PPh3)2, Na2CO3, 1,4-dioxane, 90
°C. (c) For the preparation of 24: (1) phosgene, rt; (2) methylamine, rt. For the preparation of 25: (1) phenyl chloroformate, Na2CO3, 1,4-dioxane,
rt; (2) methylamine, Et3N, DMF, 50 °C. (d) N-Methyl-4-aminophenylboronic acid pinacol ester, PdCl2(PPh3)2, Na2CO3, DMF/DME/water/
EtOH, 90 °C. (e) (1) Phenyl chloroformate, Na2CO3, 1,4-dioxane, rt; (2) methylamine, Et3N, DMF, 60 °C.
was achieved through a two-step procedure in which the
alcohol was first converted to the mesylate, followed by
reaction with sodium iodide at elevated temperature. This
two-step conversion was carried out in good yield. Treatment
of C with methanesulfinic acid sodium salt resulted in the
direct installation of the required sulfone moiety to generate
intermediate D.24 Again this conversion proceeded in good
yield and no reaction of the sulfinic acid with the 2-chloro
C
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a
Scheme 3. Synthesis of Compounds 18−22
a
Reagents: (a) [4-(methylcarbamoylamino)phenyl]boronic acid pinacol ester (prepared in situ by reaction of 1-(4-bromophenyl)-3-methylurea with
bis(pinacolato)diboron and Pd(dppf)2 in 1,4-dioxane at 90 °C), Pd(dppf)2, 1,4-dioxane, 90 °C. (b) (1) MsCl, Et3N, DCM, 0 °C; (2) appropriate
thiol, DBU, MeCN; (3) mCPBA, NaMnO4, 1,4-dioxane/water. (c) [4-(Cyclopropylcarbamoylamino)phenyl]boronic acid, Pd(PPh3)2Cl2, Na2CO3,
DMF/DME/water/EtOH, 90 °C. (d) (1) MsCl, Et3N, DCM, 0 °C; (2) 3-sulfanylpropan-1-ol, DBU, MeCN; (3) mCPBA, NaMnO4, 1,4-dioxane/
water. (e) Cyclopropylsulfinic acid sodium salt, MeCN, 90 °C. (f) (4-Aminophenyl)boronic acid pinacol ester, PdCl2(PPh3)2, Na2CO3, DMF/
DME/water/EtOH, 90 °C. (g) (1) Phenyl chloroformate, Na2CO3, 1,4-dioxane, rt; (2) methylamine, Et3N, DMF, 60 °C.
a
Scheme 4. Synthesis of Compounds 26−32
a
Reagents: (a) NaOtBu, MeI, DMF, 0 °C. (b) (4-Aminophenyl)boronic acid pinacol ester, PdCl2(PPh3)2, Na2CO3, DMF, 80 °C. (c) 1) Phenyl
chloroformate, Na2CO3, 1,4-dioxane, rt; (2) appropriate amine, Et3N, DMF, 60 °C. (d) Bu4NBr, 1,2-dibromoethane, NaOH(aq), toluene, 60 °C. (e)
(4-Aminophenyl)boronic acid pinacol ester, PdCl2(PPh3)2, Na2CO3, DMF/DME/water/EtOH, 80−95 °C. (f) Bu4NBr, 1,3-dibromopropane,
NaOH(aq), toluene, 45 °C. (g) Bu4NBr, 1,4-dibromobutpane, NaOH(aq), toluene, 60 °C. (h) NaOtBu, O(CH2CH2Br)2, DMF, 0 °C to rt.
D
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a
Scheme 5. Synthesis of Compounds 33, 35−38, 40, and 41
a
Reagents: (a) 1) 3-mercaptopropan-1-ol, DIPEA, DCM; (2) mCPBA, DCM. (b) (1) TIPSCl, imidazole, DCM; (2) Bu4NBr, 1,2-dibromoethane,
NaOH(aq), toluene, 60 °C; (3) TBAF, THF. (c) (4-Aminophenyl)boronic acid pinacol ester, PdCl2(PPh3)2, Na2CO3, DMF/DME/water/EtOH, 80
°C. (d) (1) Phenyl chloroformate, Na2CO3, 1,4-dioxane, rt; (2) cyclopropylamine, Et3N, DMF, 60 °C. (e) 1) 4-Mercaptopyridine, DIPEA, MeCN;
(2) H2O2, Na2WO4·2H2O, H2SO4, 1,4-dioxane, 55 °C. (f) 4-Fluorobenzenesulfinic acid sodium salt, MeCN, 80 °C. (g) Benzenesulfinic acid sodium
salt, MeCN, 80 °C. (h) Bu4NBr, 1,2-dibromoethane, NaOH(aq), toluene, 60−70 °C. (j) (4-Aminophenyl)boronic acid pinacol ester, PdCl2(PPh3)2,
Na2CO3, DMF/DME/water/EtOH, 80−95 °C. (k) (1) Phenyl chloroformate, Na2CO3, 1,4-dioxane, rt; (2) appropriate amine, Et3N, DMF, 60 °C.
substituent was observed. Palladium mediated cross coupling
of D with (4-aminophenyl)boronic acid pinacol ester resulted
in intermediate E which could subsequently be converted to
the required urea compounds using a range of different
chemistries. Reaction of E directly with commercially available
isocyantes delivered compounds 5, 9, 13−15 in good to
excellent yield (56−94%). Intermediate E could also be
reacted with phosgene followed by reaction with the
appropriate amine to deliver compounds 6 and 8 in a two-
step procedure, albeit with modest yields (11−38%).
Alternatively, intermediate E could be reacted with phenyl
chloroformate followed by reaction with the appropriate
amine to deliver compounds 10−12, 16, 17. Yields for this
two-step procedure varied (22−71%), and the phenyl
chloroformate intermediate formed was stable allowing
isolation and long-term storage if required. The reaction of
E with di(imidazol-1-yl)methanethione followed by reaction
with cyclopropylamine and triethylamine gave access to the
thiourea 23 in a moderate 47% yield.
The preparation of compounds 4, 24, 25, and 7 is
described in Scheme 2. Compound 4 was synthesized in
moderate yield (44%) by reaction of intermediate F, described
elsewhere,16 by reaction with ethyl isocyanate. Palladium
mediated reaction of intermediate D with the appropriate
commercially available boronic acid or boronic ester furnished
intermediates G, H, and I. Where the appropriate boronic
acid or boronic ester was not commercially available, they
could be prepared in situ from a palladium mediated
borylation of the appropraite bromide with bis(pinacolato)-
diboron. Compound 24 was prepared from intermediate G by
reaction with phosgene, followed by methylamine, whereas
compound 25 was prepared from intermediate H by reaction
with phenyl chloroformate, followed by methylamine.22
Compound 7 was prepared from intermediate I by reaction
with phenyl chloroformate followed by methylamine albeit in
a low 4% yield.
The synthesis of compounds 18−22 is described in Scheme
3.22 Palladium mediated cross coupling of intermediate B with
[4-(methylcarbamoylamino)phenyl]boronic acid pinacol ester,
made by the in situ palladium mediated reaction of 1-(4-
bromophenyl)-3-methylurea with bis(pinacolato)diboron, fur-
nished intermediate J in a modest yield (32%). A three-step
proceedure involving meslyation of the alcohol, displacement
with the appropriate thiol, and final oxidation was used to
install the sulfone substituents resulting in compounds 19, 21,
and 22 in a moderate yield (15−43%). An analogous
synthesis reacting B with the commercially available [4-
(cyclopropylcarbamoylamino)phenyl]boronic acid furnished
intermediate K in good yield (83%). The three-step
conversion of the alcohol to sulfone delivered 20 in a 17%
yield. Reaction of intermediate C with cyclopropylsulfinic acid
gave L (70% yield) which could be further functionalized by
palladium mediated cross-coupling to give intermediate M in
good yield (92%). Installation of the urea functionality by
reaction of M with phenyl chloroformate followed by reaction
with methylamine gave 18 in a reasonable 53% yield.
The synthesis of compounds 26−32 is shown in Scheme
4.22,25 Intermediate D could be alkylated twice by sequental
deprotonation/alkylation steps to furnish intermediate N in a
respectable 72% yield. Alternatively intermediate D could be
alkylated with 1,2-dibromoethane using phase transfer
conditions to give intermediate P in good yield (78%).
Phase transfer conditions could also be employed to convert
intermediate D to intermediate R (33% yield) or intermediate
E
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a
Scheme 6. Synthesis of Compounds 34, 39, and 42−44
a
Reagents: (a) (1) di(imidazol-1-yl)methanethione, DCM, rt; (2) ethanolamine, Et3N, rt. (b) (1) Di(imidazol-1-yl)methanethione, DCM, rt; (2)
appropriate amine, Et3N, rt.
T (80% yield). The lower yield for the construction of
intermediate R possibly reflects the less favorable nature of
constructing four-membered rings compared to rings of other
sizes. The tetrahydropyran ring in intermediate V was
constructed by the double deprotonation of D followed by
reaction with bis(bromoethyl) ether in good yield (78%).
Palladium mediated cross-coupling reactions were used to
convert intermediates N, P, R, T, V to intermediates O, Q, S,
U, W in moderate to good yields (50−92%). These
intermediates were converted to compounds 26−32 by
reaction with phenyl chloroformate followed by the
appropriate amine in moderate to good yields (39−75%
over two steps).
The synthesis of compounds 33−44 is outlined in Scheme
5 and Scheme 6.25,23 The hydroxypropylsulfone moiety in
intermediate X was introduced by displacement of iodide C
with 3-mercaptopropan-1-ol followed by subsequent oxidation
with mCPBA (53% yield over two steps). Protection of the
alcoholic funtionality as a triisopropylsilyl ether, cyclo-
propanation using phase transfer conditions, and subsequent
fluoride mediated desilylation gave Y in an overall yield of
54%. Y was converted to Z in a 71% yield using a palladium
mediated cross-coupling reaction, and final conversion to 33
was achieved in a 59% yield. The 4-pyridylsulfone moiety
present in intermediate AA was also introduced through a
two-step displacement/oxidation proceedure; however, in this
instance sodium tungstate and hydrogen peroxide were used
as the oxidants to limit N-oxide formation. Intermediate AA
was prepared in 79% yield from intermediate C. Reaction of
intermediate C with either 4-fluorobenzenesulfinic acid or
benzenesulfinic acid furnished intermediates AB and AC in a
76% and 85% yield, respectively. Phase transfer cyclo-
propanation was performed on intermediates L, AA, AB,
and AC to give intermediates AD, AE, AF, and AG in
excellent yield (82−93%). These intermediates could be
further functionalized by palladium mediated cross-coupling to
furnish intermediates AH, AI, AJ, and AK, again in high yield
(73−97%). Installation of the urea functionality was achieved
by a two-step procedure involving the reaction of the aniline
intermediates with phenyl chloroformate followed by amine
displacment to give compounds 35−38, 40, and 41 in good
yield (61−88% over two steps).
RESULTS AND DISCUSSION
■
Compound 3 is a potent inhibitor of mTOR with a high level
of selectivity over PI3Kα and shows cellular modulation of
both mTORC1 and mTORC2 substrates. However, the low
aqueous solubility of 3 was felt to preclude the further
development of this compound. We hypothesized that the low
solubility of 3 was driven by a combination of high
lipophilicity (log D > 3.2) together with the ability to form
strong crystal lattices due the planar nature of the molecule
and the potential to form extended hydrogen bonding
networks. The identification of compounds with improved
cellular potency and aqueous solubility, while maintaining or
improving selectivity over PI3K, became the primary focus for
the optimization campaign. To improve the chance that any
compound resulting from this optimization campaign would
be of sufficient quality to warrant clinical development, the
team aimed to equal or surpass the cellular potency and
aqueous solubility reported for the known clinical candidate
AZD8055 (cellular potencies as measured against pS6-
(S235/236) and pAKT(S473) were pIC50 = 7.57 and pIC50
=
7.62, respectively, and the aqueous solubility was measured as
30 μM).26,27
The concept of using composite parameters as a measure of
compound quality and progress within a series, such as ligand
efficiency (LE), lipophilic ligand efficiency (LipE), or
unbound clearance (Clu), is well established within the
medicinal chemistry community.28 Considering the issues
faced within this novel series of mTOR inhibitors, namely,
optimizing potency and aqueous solubility, we adopted the
composite parameter of cellular pIC50 − p(aqueous solubility
[M]) as a measure of compound progression. A large internal
data set (data not shown) has established a good correlation
for cellular potency values as measured by both the
pAKT(S473) and the pS6(S235/236) end points within this
series, and in an effort to simplify the screening cascade, only
the pAKT(S473) data were measured routinely and used to
drive chemical optimization. Cellular potency against the
pS6(S235/236) end point, alongside mTOR enzyme potency
and PI3K enzyme potency, was generated subsequently for
key compounds.
Previous groups have reported how subtle changes to the
morpholine hinge binding motif can have significant impact
on mTOR potency, selectivity over PI3K, and physicochem-
ical properties.29 In particular, work from our own laboratories
had highlighted a switch to 3(S)-methylmorpholine as being
associated with improved cellular potency and aqueous
solubility for related series.16,26 Similar effects were observed
in this novel series of urea containing compounds as
Conversion of aniline intermediates O, Q, AH, and Z to
the thioureas 34, 39, and 42−44 was achieved by initial
reaction with di(imidazol-1-yl)methanethione followed by
displacement with the appropriate amine in moderate to
excellent yield (41−99%).
F
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exemplified by compounds 4 and 5, pAKT(S473) pIC50 of 5.90
and 6.17, respectively, and aqueous solubility 17 and 120 μM,
respectively (Figure 3). It is tempting to speculate that the
Table 1. Importance of Urea Hydrogen Bond Donors
Figure 3. Structures of 4 and 5.
addition of this chiral methyl favors a more favorable binding
conformation for the molecule while in addition disrupting
the efficiency of crystal packing in the solid state by increasing
the twist between the morpholine and pyrimidine rings.
These trends are observed across a number of morpholine
vs 3(S)-methylmorpholine matched pairs within this urea
containing scaffold with an average increase in cellular pIC50
of 0.56 and an increase in aqueous solubility of approximately
3-fold (Figure 4). No attempt to characterize the solid state of
the compounds prior to assessing the aqueous solubility was
made in this analysis.
a
Data quoted are the mean pIC50 of at least three different
measurements SEM.
of the donor is vital for adopting a favorable urea
conformation for binding.
A systematic exploration of the urea substituent was
undertaken to understand the impact on cellular potency
and physicochemical properties (Table 2). Simple alkyl
substituents were tolerated (5, 6, and 9) with increasing
lipophilicity, resulting in an increase in potency alongside a
reduction in aqueous solubility. However, when analyzing LLE
or pIC50 − pSolubility, it is clear that there is a slight
reduction of compound quality as the size of the alkyl
substituent increases. A significant improvement in both
cellular potency and LLE is observed upon the introduction of
a cyclopropyl substituent, 10, perhaps reflecting an improved
interaction with the protein and the reduced lipophilicity of
this substituent compared to isopropyl. However, compound
quality as measured by pIC50 − pSolubility is reduced. The
introduction of polar functionality is tolerated as exemplified
by 11 which exhibits good potency and much improved
solubility, thus demonstrating improved quality as measured
by either LLE or pIC50 − pSolubility. Compound 11 retains
the excellent selectivity for binding to the mTOR enzyme
over the PI3Kα enzyme that is generally observed within this
series (mTOR pIC50 = 8.24, PI3Kα pIC50 = 5.49). The
introduction of a basic substituent, such as 12, significantly
lowers the log D and improves the aqueous solubility albeit
with a concomitant reduction in potency. Analyses of LLE
and pIC50 − pSolubility parameters highlight 12 as a
compound with improved quality despite the lower absolute
potency, and this compound was therefore selected for further
investigation. However, more detailed profiling revealed that
both mTOR binding affinity and selectivity over PI3Kα are
dramatically reduced following the introduction of the basic
substituent (mTOR pIC50 = 6.12, PI3Kα pIC50 = 5.10). The
reduced selectivity (as calculated from enzyme binding
affinities), alongside the unexplained difference in enzyme−
cell correlation for this compound, discouraged us from
further investigation of basic substituents in this position. The
introduction of aryl substituents to the urea is tolerated and
results in potent compounds (13−15); however, the increase
Figure 4. Matched pairs analysis highlighting the impact of switching
from morpholine to (3S)-3-methylmorpholine on both cell potency
and solubility.
The importance of the hydrogen bond donor motif in the
para-position of the 2-aryl substituent of the pyrimidine was
highlighted during the conception of the urea containing
scaffold and is further confirmed by the loss of potency upon
methylation of this nitrogen (7 cf. 6) (Table 1). In addition,
the second urea hydrogen bond donor also appears to be
important with methylation again resulting in a loss of
potency (8 cf. 6), suggesting that this nitrogen also forms a
constructive interaction with the protein or that the presence
G
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b
Table 2. Impact of Urea Substituent on Cellular Potency and Physicochemical Properties
a
b
Data quoted are the mean pIC50 of at least three different measurements SEM. The asterisk (∗) indicates evidence of crystallinity as assessed by
polarized light microscopy.
in lipophilicity combined with the reduction in aqueous
observed for the cyclohexyl substituent, 19. Analyses of the
compound quality metrics show that despite improved
potency, compound quality is eroded slightly as the size of
alkyl group is increased. Polar substituents are tolerated on
the sulfone with compound 20 showing a promising balance
of potency and solubility, albeit with a similar quality
assessment to the simple methyl substituted compound, 6.
Aryl and heteroaryl substituents are tolerated, as exemplified
by 21 and 22, and result in increased potency; however, the
lipophilic nature of these substituents results in only modest
improvements in compound quality.
The SAR investigations into the urea and sulfone portions
of the molecule highlighted the general positive relationship
between lipophilicity and cellular potency for the series in
addition to the general negative relationship between
lipophilicity and aqueous solubility. The challenge to identify
compounds with the desired balance of cellular potency and
solubility results in a reduction in both LLE and pIC50
−
pSolubility when compared with simple alkyl substituents.
The electronic nature of the substituent on the aryl ring
appears to have little impact on either cellular potency or
solubility. Heteroaryl substituents are also tolerated with 16
showing a promising balance of potency and solubility and a
compound quality similar to 6. Benzyl substituents, such as
17, are not tolerated suggesting that the conformation
adopted by such groups is incompatible with efficient binding.
Following the assessment of the urea substituent attention
was turned to the sulfone moiety and another systematic
exploration of the SAR undertaken (Table 3). Small alkyl
substituents were tolerated with a general increase in potency
and reduction in solubility observed upon increasing bulk,
exemplified by 6 and 18; however, there does appear to be a
limit to the size of the substituent with a reduction in potency
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b
Table 3. SAR of the Sulfone Substituent
a
b
Data quoted are the mean pIC50 of at least three different measurements SEM. The asterisk (∗) indicates evidence of crystallinity as assessed by
polarized light microscopy.
aqueous solubility in light of these conflicting relationships is
apparent, and it was appreciated that simple variation of these
two positions was unlikely to deliver a compound of the
required profile. We therefore sought alternative optimization
strategies to continue to improve compound quality. The
ability of a molecule to form stable lattices in the crystalline
state is known to influence the solubility of the compound
and thus potentially provides another avenue to modulate
solubility in addition to the modulation of lipophilicity. Other
groups have reported approaches targeting the ability of
molecules to form hydrogen bond networks or efficient π−π
staking interactions in the crystal lattice as a strategy to
improve aqueous solubility.30 To understand if a similar
strategy could be utilized for our series of mTOR inhibitors, a
small molecule crystal structure of 15 was obtained (Figure
5). The small molecule crystal structure highlights the
extended hydrogen bond network formed between the
hydrogen bond donors of the urea and the sulfone oxygens
on adjacent molecules. The largely planar conformation of the
molecules is also apparent thus resulting in efficient stacking.
The importance of the urea hydrogen bond donors for
mTOR potency has already been established thus invalidating
strategies that involve their removal to disrupt the hydrogen
bonding network. A variety of known urea isosteres were
investigated with the thiourea, 23, showing similar cellular
potency to the urea analogue, 10, but with slightly improved
aqueous solubility perhaps reflecting the differing hydrogen
bonding strength of thiourea versus urea, Table 4.31 All other
urea isosteres attempted displayed a disappointing loss of
potency (data not shown). Early SAR studies had established
that the sulfone oxygens are not essential for mTOR potency
(data not shown) although it was anticipated that the removal
of these oxygens would result in a significant increase in
lipophilicity, thus rendering such a strategy as unattractive at
this stage. The low probability of success associated with
strategies targeting the disruption of the hydrogen bonding
network led us to focus our efforts on disrupting the planarity
of the molecules and thus their ability to efficiently stack. The
introduction of substituents ortho to the urea did indeed
result in improved aqueous solubility, presumably through the
disruption of planarity, but a significant reduction in potency
was observed, 24 and 25 (Table 4).
Further inspection of the small molecule crystal structure
revealed the sulfone moiety to be slightly out of plane
compared to the pyrimidine ring (Figure 5). This
conformation is also present in the proposed binding mode
of 2 in a homology model of mTOR (Figure 2b). The
apparent ability for the mTOR protein to accommodate this
conformation prompted us to explore options to either
stabilize this conformation or to increase the degree of
nonplanarity in this region of the molecule. To explore this
strategy further, compound 26 was prepared in which the
methylene group linking the sulfone to the pyrimidine has
been dimethylated (Figure 6). Compound 26 shows a
significant improvement in cellular potency (pIC50 = 7.62)
when compared to compound 6, while reasonable aqueous
solubility has been maintained (46 μM) thus making this
compound the first from the series to show comparable cell
potency/aqueous solubility to AZD8055. The improved
compound quality is also reflected in both LLE (5.3) and
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Figure 6. Structure of 26.
(pIC50 = 6.29) when assessed in the appropriate enzyme
assays.
A more systematic investigation into the consequences of
alkylation on the methylene linker was performed (Table 5).
Again, dimethylation results in a potency improvement and a
significant improvement in the pIC50 − pSolubility quality
measure (27 cf. 10). A further increase in pIC50 − pSolubility
was observed upon the introduction of a cyclopropyl moiety,
28, driven mostly by an increase in aqueous solubility.
Cyclobutyl and cyclopentyl groups are tolerated, 29 and 30,
respectively, and exhibit excellent levels of cellular potency;
however, this is offset by the increased lipophilicity resulting
in a reduction in both LLE and pIC50 − pSolubility. The
introduction of a tetrahydropyranyl linker, 31, shows
promising cellular potency, but despite the reduction in
lipophilicity compared to 29 and 30, the aqueous solubility
remains low.
The increase in compound quality highlighted through
these SAR studies, and in particular the increase in pIC50
−
pSolubility identified following alkylation of the methylene
linker, has resulted in the discovery of a number of
compounds possessing cellular potency and aqueous solubility
profiles comparable with that of AZD8055 (e.g., 26, 27, and
28). The detailed assessment of AZD8055 in preparation for
clinical development highlighted a high turnover in human
hepatic incubations. The stability of compounds toward
metabolism by human hepatocytes is often used as a key
preclinical experiment to predict the likely human pharmaco-
kinetics and therefore likely clinical dose, and thus,
Figure 5. Small molecule crystal structure of 15 (CCDC 1019570)
highlighting (a) the packing scheme along the b-axis and (b)
hydrogen bonding between the urea and sulfone functionality of
neighboring molecules and the largely planar nature of the molecule.
pIC50 − pSolubility (3.3) measures. Compound 26 retains
excellent selectivity for mTOR (pIC50 = 9.01) over PI3Kα
b
Table 4. SAR of the Urea Structural Motif
a
b
Data quoted are the mean pIC50 of at least three different measurements SEM. The asterisk (∗) indicates evidence of crystallinity as assessed by
polarized light microscopy.
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Table 5. Alkylation of the Methylene Spacer
a
Data quoted are the mean pIC50 of at least three different measurements SEM.
compounds with high turnover in human hepatocyte
incubations are often considered as high risk when trying to
identify compounds with suitable properties for clinical
dosing. Human hepatocyte incubation studies performed on
26, 27, and 28 highlighted high turnover for each of these
compounds (CLint of 36.3, 50.0, and 37.8 (μL/min)/106 cells,
respectively). The optimization of stability toward metabolism
by human hepatocytes played an important role in the
preclinical decision to select AZD2014 (CLint < 4.2 (μL/
min)/106 cells) as a complementary clinical candidate to
AZD8055 (CLint = 36.4 (μL/min)/106 cells) but with
reduced human pharmacokinetic risk and is discussed
elsewhere.26 In order to identify compounds that would
have a reduced human pharmacokinetic risk for clinical
development, the team used the learning developed within
these laboratories during the discovery of AZD2014 and
decided to implement an upper limit for human hepatic
stability (CLint of <10 (μL/min)/106 cells) as a key
compound progression criteria.
Initial SAR investigations had looked to vary only a single
portion of the molecule at a time to better understand the
impact of any changes; however, in order to explore potential
interdependencies in SAR between these portions, a range of
compounds combining multiple changes were prepared
(Table 6). These combinations delivered a number of highly
potent examples and established the general additive nature of
the SAR described. While a number of the most potent
compounds showed aqueous solubility below the desired level,
such as 32−34, compounds were identified that met, or
almost met, both the desired potency and solubility
requirements (35−39). However, the majority of compounds
showed relatively high hepatic turnover with only compound
39 comfortably meeting the cellular potency, aqueous
solubility, and hepatic stability requirements. Compound 39
was shown to be highly bioavailable in mouse (F ≈ 100%)
although the compound appears to suffer from a relatively low
permeability and high efflux ratio which may constitute a risk
if treating tumors that express high levels of efflux transporters
(compound 39 shows an apparent permeability (Papp) in the
A−B direction of 0.07 × 10−6 cm/s in an MDCKII assay34
and a Papp in the B−A direction of 17 × 10−6 cm/s).
Although improvements in compound quality are apparent
following the SAR investigations described, the challenge to
identify compounds with the desired levels of potency,
aqueous solubility, and stability in human hepatic incubations
remains a significant challenge, not least because of the
apparent opposing effect of lipophilicity on these parameters.
This challenge, coupled with the large number of possible
combinations of substituents around the molecule which
would be predicted to be tolerated, prompted us to explore
the use of predictive models to more efficiently mine the
chemical space. Given the large data set already compiled
through the SAR investigations described, along with the
additive nature of the SAR, we were drawn to the use of
Free−Wilson analysis as a tool to build predictive models.32
A
Free−Wilson analysis looks to use regression analyses to
K
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Table 6. Combinations of Different Ureas, Sulfones, and Spacers
a
Data quoted are the mean pIC50 of at least three different measurements SEM.
Figure 7. Computational enumeration of chemical space.
define the contribution of specific substituents within a
molecule toward the measured property of the entire
molecule, usually biological potency. Once defined, the
substituent contribution values from different regions of the
molecule can be combined to generate a predicted value for a
molecule containing any given combination of substituents.
To use such an approach to predict the parameters of
molecules within our series, it was first necessary to define the
L
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Figure 8. Free−Wilson models for pAKT (S473) pIC50, log(Solubility), log(human Heps CLint).
Table 7. Compounds Meeting Desired Potency, Solubility, and Human Hepatic Stability Requirements
a
Data quoted are the mean pIC50 of at least three different measurements SEM.
chemical space of interest.33 A set of 21 preferred urea
substituents, 20 preferred sulfone substituents, 6 differently
substituted methylene linkers, and 2 hinge binder groups
(morpholine and 3(S)-methylmorpholine) were selected to
define the chemical space. In addition both urea and thiourea
analogues were allowed (Figure 7). Once the chemical space
solubility and human hepatic turnover appear to be of lesser
quality (as assessed by RMSE) but were still believed to be of
sufficient value for inclusion in this exercise (Figure 8).
All possible combinations of substituents were enumerated
in silico to represent the entire chemical space described
(10 080 virtual compounds) and the cellular pIC50,
log10(Solubility), and log10(CLint) calculated for each. Analysis
of this predicted data set was performed to identify those
novel combinations predicted to have the best chance of
achieving the desired balance of potency, solubility, and
hepatic stability. As a result of this exercise, 122 virtual
compounds were prioritized for synthesis. While the majority
of these compounds failed to fully meet all the desired
criteria, reflecting the significant challenge of combining these
properties within a single molecule from this scaffold as well
as reflecting the error inherent in any predicted model, a
had been defined, then the cellular potency (pAKT(S473
)
pIC50), aqueous solubility (log10[Solubility (M)]), and human
hepatic turnover data (log10[CLint ((μL/min)/106 cells)]) for
any existing compound within this space were collated and
subjected to a Free−Wilson analysis. In total the data from
494 existing compounds were computationally deconstructed
to generate substituent contribution values. The highest
quality model appears to be that for cellular potency
presumably reflecting the larger data set and the known
additive nature of the SAR. The models for the aqueous
M
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significant enrichment in compounds successfully meeting the
criteria was observed. Approximately 4% of compounds
prioritized for synthesis following this exercise met all of
the following criteria: cellular pIC50 of >7.5, solubility of >30
μM, human hepatic CLint of <10 (μL/min)/106 cells. This
compares with a ∼0.5% success rate for compounds
synthesized in the previous SAR investigations. A significant
enrichment is also seen in the number of compounds that
successfully met two of the three criteria (12% vs 3%). The
enhanced success rate observed for compounds meeting the
criteria defined is seen as validation for the use of such
models to more efficiently mine chemical space. As a result of
the process described above, compounds 40−44 were
identified as being worthy of further investigation (Table 7).
More detailed profiling of compounds 40−44, including in
vivo pharmacokinetic evaluation, was performed in order to
ascertain the suitability of the compounds as potential clinical
candidates. Compounds 43 and 44 showed poor oral
exposure in mouse (<3% bioavailability) which may be in
part due to compromised permeability and/or efflux liability
(compound 43 shows Papp(A−B) = 0.18 × 10−6 cm/s and
when tested at concentrations up to 10 μM, and shows no
activity against the hERG (human ether-a-go-go-related gene)
ion channel (IC50 > 100 μM).36
CONCLUSION
■
In summary, we have outlined the detailed SAR investigations
for a novel series of urea containing dual mTORC1 and
mTORC2 inhibitors that show excellent selectivity over
PI3Kα. Empirical SAR understanding was combined with
knowledge of molecular interactions from a small molecule
crystal structure to identify opportunities to improve solubility
without adversely impacting cellular potency. The composite
parameters of LLE and pIC50 − pSolubility were used to help
quantify compound quality and progress. The apparent
opposing relationships between lipophilicity and both cellular
potency and solubility resulted in a significant challenge to
identify compounds with the desired profile and prompted the
use of predictive models to aid compound design. Free−
Wilson analyses were used to predict cellular potency,
aqueous solubility, and human hepatocyte turnover for a
large matrix of virtual compounds. The use of these models
results in a significant enrichment in the proportion of
compounds synthesized that successfully met the desired
criteria and thus represents a more efficient strategy for
mining the chemical space defined. These studies culminated
in the identification of AZD3147 (42), which is an extremely
potent and selective dual inhibitor of mTORC1 and
mTORC2 and possesses physicochemical, pharmacokinetic,
and other properties suitable for development as a potential
clinical candidate.
P
app(B−A) = 22 × 10−6 cm/s in an MDCKII assay;
compound 44 was not measured in this assay but has further
reduced log D and an increased hydrogen bond donor count
both of which are known to impair permeability).
Compounds 40 and 41 show moderate to good oral
exposures in mouse (35% and 55% bioavailability, respec-
tively) but only moderate oral exposures in dog (29% and
21% bioavailability, respectively).35 Permeability analysis in the
MDCKII assay again highlighted low passive permeability and
an efflux liability for these compounds (Papp(A−B) = 0.18 ×
10−6 cm/s and Papp(B−A) = 24 × 10−6 cm/s for compound
40; Papp(A−B) = 0.15 × 10−6 cm/s and Papp(B−A) = 31 ×
10−6 cm/s for compound 41). Compound 42, however,
displays an improved permeability/efflux profile in the
MDCKII assay (Papp(A−B) = 5.6 × 10−6 cm/s and Papp(B−
A) = 43 × 10−6 cm/s) and was found to give good oral
bioavailability in both mouse and dog (61% and 73%,
respectively). The improved bioavailability of 42 together with
its encouraging physicochemical properties resulted in the
selection of this compound for preclinical development, and it
was subsequently given the internal identifier of AZD3147. A
more detailed description of its properties is given in Table 8.
AZD3147 (42) is a potent inhibitor of mTOR with a high
degree of selectivity over the PI3K isoforms and against a
general panel of kinases (only 2 hits with >50% inhibition
when tested at 10 μM against a panel of 98 kinases).
AZD3147 (42) inhibits both mTORC1 and mTORC2
substrates in cells, does not appear to inhibit CYP enzymes
MATERIALS AND METHODS
■
General Methods. All experiments were carried out under an
inert atmosphere and at room temperature unless otherwise stated.
Microwave reactions were performed using one of the following
reactors: Biotage initiator, Personal Chemistry Emrys optimizer,
Personal Chemistry Smithcreator, or CEM Explorer. Workup
procedures were carried out using traditional phase separating
techniques or by using strong cation exchange (SCX) chromatog-
raphy using Isolute SPE flash SCX-2 column (Internationsl Sorbent
Technology Limited, Mid Glamorgan, U.K.). When necessary,
organinc solvents were dried over anhydrous MgSO4 or Na2SO4.
Flash column chromatography was performed on Merck Kieselgel
silica (article 9385) or on reversed phase silica (Fluka silica gel 90
C18) or on Silicycle cartridges (40−63 μm silica, 4−330 g weight)
or on Grace resolv cartridges (4−120 g) or on RediSep Rf 1.5 Flash
columns or on RediSep Rf high performance Gold Flash columns
(150−415 g weight) or on RediSep Rf Gold C18 reversed-phase
columns (20−40 μm silica) either manually or automated using an
Isco Combi Flash Companion system or similar system. Preparative
reverse phase HPLC (RP HPLC) was performed on C18 reversed-
phase silica, for example, on a Waters “Xterra” or “XBridge”
preparative reversed-phase column (5 μm silica, 19 mm diameter,
100 mm length) or on a Phenomenex “Gemini” or “AXIA”
preparative reversed-phase column (5 μm silica, 110A, 21.1 mm
diameter, 100 mm length) using decreasingly polar mixtures as
eluent, for example [containing 0.1−5% formic acid or 1−5%
aqueous ammonium hydroxide (d = 0.88)] as solvent A and
acetonitrile as solvent B or MeOH/MeCN 3:1. A typical procedure
would be as follows: a solvent gradient over 9.5 min, at 25 mL/min,
from a 85:15 (or alternative ratio as appropriate) mixture of solvents
A and B, respectively, to a 5:95 mixture of solvents A and B.
Intermediates were not necessarily purified, but their structures and
purity were assessed by TLC, NMR, HPLC, and mass spectral
techniques and are consistent with the proposed structures. The
purity of compounds for biological testing was assessed by NMR,
Table 8. Profile of 42 (AZD3147)
42 (AZD3147)
mTOR pIC50
8.82
PI3Kα, -β, -δ, -γ pIC50
pAkt pIC50, pS6 pIC50
fu (m, r, d, h)37
6.04, 5.26, 5.03, 5.20
8.24, 7.39
0.29, 0.26, >0.67, 0.41
78, 2.3, 0.9, 61
16, 2.0, 1.9, 73
5.6, 43
a
mouse (CL, Vss, t1/2, F)
a
dog (CL, Vss, t1/2, F)
b
MDCK Papp(A−B), Papp(B−A)
a
Units for CL, Vss, t1/2, and F are mL/min/kg, L/kg, h, and %,
b
respectively. Unit for Papp is ×10−6 cm/s.
N
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HPLC, and mass spectral techniques and are consistent with the
proposed structures; purity was ≥95%. Electrospray mass spectral
data were obtained using a Waters ZMD or Waters ZQ LC/mass
spectrometer acquiring both positive and negative ion data, and
generally, only ions relating to the parent structure are reported.
Proton NMR chemical shift values were measured on the δ scale
using a Bruker DPX300 spectrometer operating at a field strength of
300 MHz, a Bruker DRX400 operating at 400 MHz, a Bruker
DRX500 operating at 500 MHz, or a Bruker AV700 operating at 700
MHz. Unless otherwise stated, NMR spectra were obtained at 400
MHz in deuterated dimethyl sulfoxide. The following abbreviations
have been used: s, singlet; d, doublet; t, triplet; q, quartet; m,
multiplet; br, broad; qn, quintet.
J = 8.76 Hz, 2H), 8.72 (s, 1H), 8.89 (s, 1H). HRMS-ESI (m/z): [M
+ H]+ calcd for C24H26FN5O4S, 500.176 18; found 500.176 23.
1-(4-Methoxyphenyl)-3-[4-[4-[(3S)-3-methylmorpholin-4-yl]-6-
(methylsulfonylmethyl)pyrimidin-2-yl]phenyl]urea (15). Yield 56%.
1H NMR (DMSO-d6) δ 1.24 (d, J = 6.79 Hz, 3H), 3.20 (s, 3H),
3.21−3.25 (m, 1H), 3.49 (td, J = 3.06, 11.80, 11.81 Hz, 1H), 3.64
(dd, J = 3.08, 11.48 Hz, 1H), 3.71 (s, 3H), 3.77 (d, J = 11.41 Hz,
1H), 3.98 (dd, J = 3.69, 11.36 Hz, 1H), 4.16 (s, 1H), 4.48 (s, 3H),
6.77 (s, 1H), 6.83−6.91 (m, 2H), 7.31−7.43 (m, 2H), 7.49−7.59 (m,
2H), 8.19−8.3 (m, 2H), 8.49 (s, 1H), 8.81 (s, 1H). 13C NMR
(DMSO, d6) 13.25, 38.62, 41.37, 46.35, 55.13, 61.43, 65.97, 70.12,
102.00, 113.96, 117.34, 120.12, 128.47, 130.70, 132.43, 142.18,
152.43, 154.57, 157.32, 161.69, 162.24. HRMS-ESI (m/z): [M + H]+
calcd for C25H29N5O5S, 512.196 22; found, 512.196 04.
The following abbreviations may be used: THF = tetrahydrofuran;
DIPEA = diisopropylethylamine; DMF = N,N dimethylformamide;
DMSO = dimethyl sulfoxide; DMA = N,N-dimethylacetamide; DME
= 1,2-dimethoxyethane; DCM = dichloromethane; MeOH =
methanol;
Compounds 6 and 8 were prepared from intermediate E
according to the general procedure B. Compound 24 was prepared
from intermediate G according to the general procedure B.
General Proceedure B.
A solution of the appropriate
intermediate (1 equiv) in a mixture of DCM and pyridine (4:1)
was added dropwise to a solution of phosgene (20% solution in
toluene, 1.2 equiv) in DCM and the reaction stirred for 1 h at
ambient temperature. The appropriate amine (1.2 equiv) was added
and the reaction stirred for a further 1 h or until the reaction was
complete by TLC or LCMS analysis. The reaction was partitioned
between ethyl acetate and water, and the organics were dried over
MgSO4 and concentrated in vacuo. The crude material was purified
by either flash silica chromatography or HPLC.
1-Methyl-3-[4-[4-[(3S)-3-methylmorpholin-4-yl]-6-
(methylsulfonylmethyl)pyrimidin-2-yl]phenyl]urea (6). Yield 38%.
1H NMR (DMSO-d6) δ 1.25 (d, J = 6.72 Hz, 3H), 2.67 (d, J = 4.64
Hz, 3H), 3.21 (s, 3H), 3.23 (m, 1H), 3.44−3.6 (m, 2H), 3.66 (dd, J
= 2.95, 11.52 Hz, 1H), 3.78 (d, J = 11.47 Hz, 1H), 3.99 (dd, J =
3.47, 11.40 Hz, 1H), 4.18 (d, J = 11.23 Hz, 1H), 4.49 (s, 3H), 6.08
(d, J = 4.66 Hz, 1H), 6.78 (s, 1H), 7.51 (d, J = 8.87 Hz, 2H), 8.21
(d, J = 8.84 Hz, 2H), 8.76 (s, 1H). HRMS-ESI (m/z): [M + H]+
calcd for C19H25N5O4S, 420.170 00; found 420.169 89.
1,1-Dimethyl-3-[4-[4-[(3S)-3-methylmorpholin-4-yl]-6-
(methylsulfonylmethyl)pyrimidin-2-yl]phenyl]urea (8). Yield 11%.
1H NMR (DMSO-d6) δ 1.25 (d, J = 6.73 Hz, 3H), 2.96 (s, 6H),
3.21 (s, 3H), 3.28 (s, 1H), 3.51 (td, J = 2.84, 11.76, 11.84 Hz, 1H),
3.66 (dd, J = 2.99, 11.46 Hz, 1H), 3.79 (d, J = 11.47 Hz, 1H), 4.00
(dd, J = 3.41, 11.49 Hz, 1H), 4.18 (s, 1H), 4.49 (s, 3H), 6.79 (s,
1H), 7.60 (d, J = 8.88 Hz, 2H), 8.21 (d, J = 8.84 Hz, 2H), 8.50 (s,
1H). HRMS-ESI (m/z): [M + H]+ calcd for C20H27N5O4S,
434.185 61; found 434.185 65.
3-[2-Methoxy-4-[4-[(3S)-3-methylmorpholin-4-yl]-6-
(methylsulfonylmethyl)pyrimidin-2-yl]phenyl]-1-methylurea (24).
Yield 30%. 1H NMR (DMSO-d6) δ 1.23 (d, J = 6.76 Hz, 3H),
2.64 (d, J = 4.58 Hz, 3H), 3.20 (m, 4H), 3.49 (td, J = 3.04, 11.81,
11.84 Hz, 1H), 3.64 (dd, J = 3.06, 11.47 Hz, 1H), 3.76 (d, J = 11.42
Hz, 1H), 3.90 (s, 3H), 3.97 (dd, J = 3.63, 11.36 Hz, 1H), 4.17 (s,
1H), 4.45 (d, J = 15.08 Hz, 1H), 4.48 (s, 2H), 6.76 (s, 1H), 6.83 (d,
J = 4.61 Hz, 1H), 7.83−7.93 (m, 2H), 8.08 (s, 1H), 8.21 (d, J = 8.40
Hz, 1H). HRMS-ESI (m/z): [M + H]+ calcd for C20H27N5O5S,
450.180 57; found 450.180 45.
Compound 7 was prepared from intermediate I according to the
general procedure C. Compounds 10, 11, 12, 16, and 17 were
prepared from intermediate E according to the general procedure C.
Compound 18 was prepared from intermediate M according to the
general procedure C. Compound 25 was prepared from intermediate
H according to the general procedure C. Compounds 26−31 were
prepared from intermediate O according to the general procedure C.
Compounds 32, 33, 36, and 37 were prepared from intermediates R,
Z, AI, and AJ, respectively, according to the general procedure C.
Compounds 35 and 38 were prepared from intermediate AH
according to the general procedure C. Compounds 40 and 41 were
prepared from intermediate AK according to the general procedure
C.
Compound 4 was prepared from intermediate F according to the
general procedure A. Compounds 5, 9, 13, 14, and 15 were prepared
from intermediate E according to the general procedure A.
General Proceedure A. The appropriate isocyanate (approx-
imately 5 equiv) was added to the appropriate intermediate (1 equiv)
in 1,4-dioxane and heated at 70 °C for 4 h or until the reaction was
complete by TLC or LCMS analysis. The reaction mixture was
concentrated in vacuo and the resultant material purified by either
flash silica chromatography or HPLC.
1-Ethyl-3-[4-[4-(methylsulfonylmethyl)-6-morpholin-4-ylpyrimi-
1
din-2-yl]phenyl]urea (4). Yield 44%. H NMR (DMSO-d6) δ 1.07 (t,
J = 7.16, 7.16 Hz, 3H), 3.13 (dd, J = 5.61, 7.15 Hz, 2H), 3.21 (s,
3H), 3.66−3.75 (m, 8H), 4.48 (s, 2H), 6.19 (t, J = 5.55, 5.55 Hz,
1H), 6.81 (s, 1H), 7.50 (d, J = 8.86 Hz, 2H), 8.21 (d, J = 8.80 Hz,
2H), 8.69 (s, 1H). HRMS-ESI (m/z): [M + H]+ calcd for
C19H25N5O4S, 420.170 00; found 420.169 92.
1-Ethyl-3-[4-[4-[(3S)-3-methylmorpholin-4-yl]-6-
(methylsulfonylmethyl)pyrimidin-2-yl]phenyl]urea (5). Yield 94%.
1H NMR (DMSO-d6) δ 0.98 (t, J = 7.16, 7.16 Hz, 3H), 1.25 (d, J =
6.71 Hz, 3H), 3.00 (qd, J = 5.66, 7.17, 7.20, 7.20 Hz, 2H), 3.08−
3.18 (m, 2H), 3.21 (s, 3H), 3.44−3.56 (m, 1H), 3.66 (d, J = 8.42
Hz, 1H), 3.74−3.82 (m, 1H), 3.99 (d, J = 8.11 Hz, 1H), 4.16 (s,
1H), 4.49 (s, 2H), 6.17 (t, J = 5.55, 5.55 Hz, 1H), 6.78 (s, 1H), 7.50
(d, J = 8.85 Hz, 2H), 8.21 (d, J = 8.83 Hz, 2H), 8.67 (s, 1H).
HRMS-ESI (m/z): [M + H]+ calcd for C20H27N5O4S, 434.185 65;
found 434.185 61.
3-[4-[4-[(3S)-3-Methylmorpholin-4-yl]-6-(methylsulfonylmethyl)-
1
pyrimidin-2-yl]phenyl]-1-propan-2-ylurea (9). Yield 66%. H NMR
(DMSO-d6) δ 1.10 (d, J = 6.52 Hz, 6H), 1.23 (d, J = 6.78 Hz, 3H),
3.19 (m, 4H), 3.48 (td, J = 3.03, 11.82, 11.84 Hz, 1H), 3.64 (d, J =
3.03 Hz, 1H), 3.71−3.82 (m, 2H), 3.97 (dd, J = 3.64, 11.35 Hz,
1H), 4.15 (s, 1H), 4.46 (s, 3H), 6.05 (d, J = 7.51 Hz, 1H), 6.75 (s,
1H), 7.41−7.52 (d, J = 8.76 Hz, 2H), 8.19 (d, J = 8.76 Hz, 2H),
8.52 (s, 1H). HRMS-ESI (m/z): [M + H]+ calcd for C21H29N5O4S,
448.201 30; found, 448.201 02.
1-[4-[4-[(3S)-3-Methylmorpholin-4-yl]-6-(methylsulfonylmethyl)-
pyrimidin-2-yl]phenyl]-3-phenylurea (13). Yield 70%. 1H NMR
(DMSO-d6) δ 1.24 (d, J = 6.80 Hz, 3H), 3.20 (s, 3H), 3.21 (m, 1H),
3.49 (td, J = 3.06, 11.77, 11.81 Hz, 1H), 3.64 (dd, J = 3.07, 11.47
Hz, 1H), 3.77 (d, J = 11.41 Hz, 1H), 3.98 (dd, J = 3.68, 11.35 Hz,
1H), 4.16 (s, 1H), 4.48 (s, 3H), 6.78 (s, 1H), 6.94−7.03 (m, 1H),
7.21−7.32 (m, 2H), 7.38−7.5 (m, 2H), 7.51−7.62 (m, 2H), 8.2−
8.31 (m, 2H), 8.69 (s, 1H), 8.89 (s, 1H). HRMS-ESI (m/z): [M +
H]+ calcd for C24H27N5O4S, 482.185 65; found, 482.185 55.
3-(4-Fluorophenyl)-1-[4-[4-[(3S)-3-methylmorpholin-4-yl]-6-
(methylsulfonylmethyl)pyrimidin-2-yl]phenyl]urea (14). Yield 65%.
1H NMR (DMSO-d6) δ 1.24 (d, J = 6.79 Hz, 3H), 3.20 (s, 3H),
3.23 (dd, J = 3.75, 12.96 Hz, 1H), 3.43−3.55 (m, 1H), 3.64 (dd, J =
3.10, 11.47 Hz, 1H), 3.77 (d, J = 11.41 Hz, 1H), 3.98 (dd, J = 3.73,
11.36 Hz, 1H), 4.16 (s, 1H), 4.48 (s, 3H), 6.78 (s, 1H), 7.12 (t, J =
8.88, 8.88 Hz, 2H), 7.42−7.52 (m, 2H), 7.52−7.61 (m, 2H), 8.25 (d,
General Proceedure C. Phenyl chloroformate (1 equiv) and
sodium carbonate (1.5 equiv) were added to a solution of the
O
DOI: 10.1021/jm501778s
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Journal of Medicinal Chemistry
Article
appropriate intermediate (1 equiv) in 1,4-dioxane, and the mixture
was stirred at ambient temperature for 2 h or until the reaction was
complete by TLC or LCMS analysis. The reaction mixture was
partitioned between ethyl acetate and water, and the organics were
dried over MgSO4 and concentrated in vacuo. The crude phenyl
carbamate was used either directly in the subsequent step or
following purification by flash silica chromatography. A solution of
the phenyl carbamate (1 equiv), the appropriate amine (2−5 equiv),
and triethylamine (3.25 equiv) in either NMP or DMF was heated
between 50 and 70 °C for 2 h or until complete by TLC or LCMS
analysis. The reaction mixture was purified by either flash column
chromatography or HPLC.
J = 0.63 Hz, 1H), 7.55 (d, J = 8.84 Hz, 2H), 7.76 (s, 1H), 8.18−8.31
(m, 2H), 8.39 (s, 1H), 8.84 (s, 1H). HRMS-ESI (m/z): [M + H]+
calcd for C22H27N7O4S, 486.191 80; found 486.191 80.
1-[4-[4-(Cyclopropylsulfonylmethyl)-6-[(3S)-3-methylmorpholin-
1
4-yl]pyrimidin-2-yl]phenyl]-3-methylurea (18). Yield 53%. H NMR
(DMSO-d6) δ 0.93−1.02 (m, 2H), 1.05 (dd, J = 2.67, 7.98 Hz, 2H),
1.23 (d, J = 6.78 Hz, 3H), 2.65 (d, J = 4.56 Hz, 3H), 2.81−2.9 (m,
1H), 3.13−3.26 (m, 1H), 3.49 (td, J = 3.09, 11.78, 11.82 Hz, 1H),
3.64 (dd, J = 3.10, 11.48 Hz, 1H), 3.76 (d, J = 11.41 Hz, 1H), 3.97
(dd, J = 3.73, 11.38 Hz, 1H), 4.45 (s, 1H), 4.48 (s, 2H), 6.05 (d, J =
4.63 Hz, 1H), 6.75 (s, 1H), 7.48 (d, J = 8.79 Hz, 1H), 8.20 (d, J =
8.81 Hz, 1H), 8.71 (s, 1H). HRMS-ESI (m/z): [M + H]+ calcd for
C21H27N5O4S, 446.185 65; found 446.185 58.
3-Methyl-1-[2-methyl-4-[4-[(3S)-3-methylmorpholin-4-yl]-6-
(methylsulfonylmethyl)pyrimidin-2-yl]phenyl]urea (25). Yield 24%.
1H NMR (DMSO-d6) δ 1.23 (d, J = 6.76 Hz, 3H), 2.24 (s, 3H),
2.66 (d, J = 4.55 Hz, 3H), 3.13−3.25 (m, 4H), 3.45−3.52 (m, 1H),
3.64 (dd, J = 3.08, 11.49 Hz, 1H), 3.76 (d, J = 11.42 Hz, 1H), 3.97
(dd, J = 3.67, 11.37 Hz, 1H), 4.17 (s, 1H), 4.47 (s, 3H), 6.54 (d, J =
4.62 Hz, 1H), 6.75 (s, 1H), 7.77 (s, 1H), 8.00 (m, 1H), 8.05 (m,
1H), 8.07 (m, 1H). HRMS-ESI (m/z): [M + H]+ calcd for
C20H27N5O4S, 434.185 65; found 434.185 61.
1,3-Dimethyl-1-[4-[4-[(3S)-3-methylmorpholin-4-yl]-6-
1
(methylsulfonylmethyl)pyrimidin-2-yl]phenyl]urea (7). Yield 4%. H
NMR (CDCl3) δ 1.37 (d, J = 6.82 Hz, 3H), 2.76 (d, J = 4.61 Hz,
3H), 3.08 (s, 3H), 3.32 (s, 3H), 3.37 (dd, J = 3.89, 13.00 Hz, 1H),
3.60 (td, J = 3.04, 11.83, 11.96 Hz, 1H), 3.75 (dd, J = 3.09, 11.52
Hz, 1H), 3.84 (d, J = 11.53 Hz, 1H), 4.06 (dd, J = 3.75, 11.57 Hz,
1H), 4.16 (d, J = 12.33 Hz, 1H), 4.27 (s, 2H), 4.36 (d, J = 4.52 Hz,
1H), 4.50 (s, 1H), 6.52 (s, 1H), 7.32 (d, J = 8.56 Hz, 2H), 8.39 (d, J
= 8.53 Hz, 2H). Mass calculated for C20H27N5O4S 433.53, found 434
(ESI, M + H).
3-Cyclopropyl-1-[4-[4-[(3S)-3-methylmorpholin-4-yl]-6-
(methylsulfonylmethyl)pyrimidin-2-yl]phenyl]urea (10). Yield 39%.
1H NMR (DMSO,d6) δ 0.41 (dd, J = 2.09, 3.75 Hz, 2H), 0.63 (dd, J
3-Methyl-1-[4-[4-[(3S)-3-methylmorpholin-4-yl]-6-(2-methylsul-
fonylpropan-2-yl)pyrimidin-2-yl]phenyl]urea (26). Yield 70%.1H
NMR (DMSO-d6) δ 1.23 (d, J = 6.72 Hz, 3H), 1.77 (d, J = 2.27
Hz, 6H), 2.66 (d, J = 4.61 Hz, 3H), 3.03 (s, 3H), 3.16−3.28 (m,
1H), 3.45−3.55 (m, 1H), 3.57−3.69 (m, 1H), 3.77 (d, J = 11.44 Hz,
1H), 3.98 (dd, J = 3.41, 11.40 Hz, 1H), 4.23 (d, J = 12.74 Hz, 1H),
4.59 (s, 1H), 6.07 (q, J = 4.45, 4.45, 4.48 Hz, 1H), 6.74 (s, 1H), 7.51
(d, J = 8.84 Hz, 2H), 8.23 (d, J = 8.80 Hz, 2H), 8.74 (s, 1H).
HRMS-ESI (m/z): [M + H]+ calcd for C21H29N5O4S, 448.201 30;
found 448.201 26.
= 1.98, 6.85 Hz, 2H), 1.23 (d, J = 6.77 Hz, 3H), 2.55 (dd, J = 3.67,
6.86 Hz, 1H), 3.19 (s, 3H), 3.22 (dd, J = 3.72, 12.99 Hz, 1H), 3.48
(td, J = 3.06, 11.78, 11.82 Hz, 1H), 3.64 (dd, J = 3.09, 11.49 Hz,
1H), 3.76 (d, J = 11.43 Hz, 1H), 3.97 (dd, J = 3.72, 11.38 Hz, 1H),
4.16 (s, 1H), 4.47 (m, 3H), 6.42 (d, J = 2.56 Hz, 1H), 6.76 (s, 1H),
7.49 (d, J = 8.79 Hz, 2H), 8.20 (d, J = 8.78 Hz, 2H), 8.52 (s, 1H).
13C NMR (DMSO-d6) 6.37, 13.30, 22.40, 38.65, 41.43, 46.40, 61.50,
66.04, 70.19, 101.98, 117.14, 128.43, 130.30, 142.70, 155.73, 157.35,
3-Cyclopropyl-1-[4-[4-[(3S)-3-methylmorpholin-4-yl]-6-(2-meth-
ylsulfonylpropan-2-yl)pyrimidin-2-yl]phenyl]urea (27). Yield 68%.
1H NMR (DMSO-d6) δ 0.40 (dd, J = 2.30, 3.64 Hz, 2H), 0.63 (dd, J
161.75, 162.37. HRMS-ESI (m/z): [M
+
H]+ calcd for
C21H27N5O4S, 446.185 65; found 446.185 58.
= 1.94, 6.85 Hz, 2H), 1.22 (d, J = 6.77 Hz, 3H), 1.76 (d, J = 4.19
Hz, 6H), 2.55 (dd, J = 3.61, 6.88 Hz, 1H), 3.02 (s, 3H), 3.16−3.25
(m, 1H), 3.48 (td, J = 3.00, 11.82, 11.85 Hz, 1H), 3.63 (dd, J = 3.02,
11.47 Hz, 1H), 3.75 (d, J = 11.40 Hz, 1H), 3.96 (dd, J = 3.59, 11.36
Hz, 1H), 4.21 (d, J = 11.41 Hz, 1H), 4.58 (s, 1H), 6.41 (s, 1H), 6.72
(s, 1H), 7.49 (d, J = 8.74 Hz, 2H), 8.22 (d, J = 8.74 Hz, 2H), 8.52
(s, 1H). 13C NMR (DMSO-d6) δ 6.30, 13.19, 20.43, 22.34, 36.41,
38.70, 46.23, 66.02, 66.39, 70.21, 99.08, 117.12, 128.38, 130.43,
142.61, 155.67, 161.48, 161.99, 164.51. HRMS-ESI (m/z): [M + H]+
calcd for C23H31N5O4S, 474.216 95; found 474.216 89.
3-Cyclopropyl-1-[4-[4-[(3S)-3-methylmorpholin-4-yl]-6-(1-
methylsulfonylcyclopropyl)pyrimidin-2-yl]phenyl]urea (28). Yield
75%. 1H NMR (DMSO-d6) δ 0.40 (dd, J = 2.15, 3.76 Hz, 2H),
0.63 (dd, J = 1.98, 6.86 Hz, 2H), 1.22 (d, J = 6.77 Hz, 3H), 1.54 (d,
J = 2.31 Hz, 2H), 1.6−1.7 (m, 2H), 2.55 (dd, J = 3.64, 6.88 Hz,
1H), 3.14−3.24 (m, 1H), 3.32 (s, 3H), 3.4−3.52 (m, 1H), 3.62 (dd,
J = 3.08, 11.48 Hz, 1H), 3.74 (d, J = 11.42 Hz, 1H), 3.95 (dd, J =
3.67, 11.38 Hz, 1H), 4.19 (s, 1H), 4.55 (s, 1H), 6.41 (d, J = 2.54 Hz,
1H), 6.75 (s, 1H), 7.49 (d, J = 8.76 Hz, 2H), 8.18 (d, J = 8.76 Hz,
2H), 8.53 (s, 1H). 13C NMR (DMSO-d6) 6.32, 12.10, 13.27, 22.34,
38.65, 39.99, 40.45, 46.02, 66.01, 70.18, 100.43, 117.12, 128.39,
130.33, 142.63, 155.68, 161.55, 161.80, 162.11. HRMS-ESI (m/z):
[M + H]+ calcd for C23H29N5O4S, 472.201 30; found 472.201 20.
3-Cyclopropyl-1-[4-[4-[(3S)-3-methylmorpholin-4-yl]-6-(1-
methylsulfonylcyclobutyl)pyrimidin-2-yl]phenyl]urea (29). Yield
50%. 1H NMR (DMSO-d6) δ 0.40 (dd, J = 2.00, 3.84 Hz, 2H),
0.63 (dd, J = 1.98, 6.89 Hz, 2H), 1.22 (d, J = 6.77 Hz, 3H), 1.83−
1.96 (m, 1H), 1.98−2.13 (m, 1H), 2.54 (dt, J = 3.42, 3.42, 6.93 Hz,
1H), 2.71−2.84 (m, 2H), 2.85 (s, 3H), 2.86−2.95 (m, 2H), 3.20 (td,
J = 3.88, 12.92, 13.04 Hz, 1H), 3.48 (td, J = 3.07, 11.82, 11.83 Hz,
1H), 3.63 (dd, J = 3.09, 11.49 Hz, 1H), 3.75 (d, J = 11.42 Hz, 1H),
3.96 (dd, J = 3.70, 11.37 Hz, 1H), 4.22 (s, 1H), 4.55 (s, 1H), 6.41
(d, J = 2.64 Hz, 1H), 6.69 (s, 1H), 7.49 (d, J = 8.77 Hz, 2H), 8.20
(d, J = 8.77 Hz, 2H), 8.53 (s, 1H). 13C NMR (DMSO-d6) 6.31,
13.23, 15.16, 22.34, 27.30, 27.35, 35.36, 38.70, 46.28, 66.03, 68.60,
3-(2-Hydroxyethyl)-1-[4-[4-[(3S)-3-methylmorpholin-4-yl]-6-
(methylsulfonylmethyl)pyrimidin-2-yl]phenyl]urea (11). Yield 22%.
1H NMR (DMSO-d6) δ 1.24 (d, J = 6.74 Hz, 3H), 3.13−3.24 (m,
5H), 3.48 (dd, J = 3.87, 15.61 Hz, 3H), 3.65 (dd, J = 2.92, 11.55 Hz,
1H), 3.78 (d, J = 11.28 Hz, 1H), 3.91−4.03 (m, 1H), 4.16 (s, 1H),
4.48 (s, 3H), 4.73 (s, 1H), 6.27 (t, J = 5.62, 5.62 Hz, 1H), 6.77 (s,
1H), 7.49 (d, J = 8.87 Hz, 2H), 8.21 (d, J = 8.81 Hz, 2H), 8.82 (s,
1H). Mass calculated for C20H27N5O5S 449.53, found 449.97 (ESI,
M + H).
3-(2-Dimethylaminoethyl)-1-[4-[4-[(3S)-3-methylmorpholin-4-
yl]-6-(methylsulfonylmethyl)pyrimidin-2-yl]phenyl]urea (12). Yield
1
44%. H NMR (DMSO-d6) δ 1.23 (d, J = 6.76 Hz, 3H), 2.17 (s,
6H), 2.32 (t, J = 6.19, 6.19 Hz, 2H), 3.14−3.23 (m, 6H), 3.48 (td, J
= 3.02, 11.87, 11.89 Hz, 1H), 3.63 (dd, J = 3.01, 11.47 Hz, 1H), 3.76
(d, J = 11.41 Hz, 1H), 3.97 (dd, J = 3.56, 11.38 Hz, 1H), 4.15 (s,
1H), 4.46 (s, 2H), 6.15 (d, J = 5.27 Hz, 1H), 6.75 (s, 1H), 7.44−
7.52 (m, J = 8.76 Hz, 2H), 8.19 (d, J = 8.76 Hz, 2H), 8.88 (s, 1H).
HRMS-ESI (m/z): [M + H]+ calcd for C22H32N6O4S, 477.227 85;
found 477.227 91.
3-[(4-Methoxyphenyl)methyl]-1-[4-[4-[(3S)-3-methylmorpholin-
4-yl]-6-(methylsulfonylmethyl)pyrimidin-2-yl]phenyl]urea (17).
Yield 71%. 1H NMR (DMSO-d6) δ 1.23 (d, J = 6.77 Hz, 3H),
3.19 (s, 3H), 3.21 (dd, J = 3.66, 12.96 Hz, 1H), 3.44−3.53 (m, 1H),
3.63 (dd, J = 3.09, 11.50 Hz, 1H), 3.72 (s, 3H), 3.76 (d, J = 11.43
Hz, 1H), 3.97 (dd, J = 3.72, 11.37 Hz, 1H), 4.15 (s, 1H), 4.23 (d, J
= 5.83 Hz, 2H), 4.46 (m, 3H), 6.58 (s, 1H), 6.75 (s, 1H), 6.89 (d, J
= 8.67 Hz, 2H), 7.23 (d, J = 8.68 Hz, 2H), 7.49 (d, J = 8.84 Hz,
2H), 8.20 (d, J = 8.79 Hz, 2H), 8.74 (s, 1H). HRMS-ESI (m/z): [M
+ H]+ calcd for C26H31N5O5S, 526.211 87; found 526.212 28.
1-[4-[4-[(3S)-3-Methylmorpholin-4-yl]-6-(methylsulfonylmethyl)-
pyrimidin-2-yl]phenyl]-3-(1-methylpyrazol-4-yl)urea (16). Yield
1
30%. H NMR (DMSO-d6) δ 1.25 (d, J = 6.73 Hz, 3H), 3.21 (s,
3H), 3.27 (s, 1H), 3.51 (td, J = 2.93, 11.78, 11.85 Hz, 1H), 3.66 (dd,
J = 2.92, 11.51 Hz, 1H), 3.79 (m, 4H), 3.99 (dd, J = 3.38, 11.32 Hz,
1H), 4.18 (d, J = 12.17 Hz, 1H), 4.49 (s, 3H), 6.79 (s, 1H), 7.38 (d,
P
DOI: 10.1021/jm501778s
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Journal of Medicinal Chemistry
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70.19, 99.90, 117.10, 128.38, 130.42, 142.62, 155.68, 161.73, 161.87,
164.38. HRMS-ESI (m/z): [M + H]+ calcd for C24H31N5O4S,
486.216 95; found 486.216 89.
(d, J = 11.49 Hz, 1H), 3.96 (d, J = 8.03 Hz, 1H), 4.02−4.19 (m,
3H), 4.73 (t, J = 4.98, 4.98 Hz, 1H), 6.22 (t, J = 5.51, 5.51 Hz, 1H),
6.64 (s, 1H), 7.3−7.51 (m, 4H), 7.69−7.95 (m, 4H), 8.78 (s, 1H).
HRMS-ESI (m/z): [M + H]+ calcd for C27H30FN5O5S, 556.202 44;
found 556.202 15.
3-Methyl-1-[4-[4-[(3S)-3-methylmorpholin-4-yl]-6-(1-pyridin-4-
ylsulfonylcyclopropyl)pyrimidin-2-yl]phenyl]urea (37). Yield 62%.
1H NMR (DMSO-d6) δ 1.17 (d, J = 6.78 Hz, 3H), 1.65−1.7 (m,
2H), 1.95 (q, J = 4.05, 4.21, 4.21 Hz, 2H), 2.64 (d, J = 4.68 Hz,
3H), 3.11−3.19 (m, 1H), 3.44 (td, J = 3.06, 11.83, 11.85 Hz, 1H),
3.59 (dd, J = 3.10, 11.48 Hz, 1H), 3.73 (d, J = 11.42 Hz, 1H), 3.94
(dd, J = 3.72, 11.39 Hz, 1H), 4.14 (s, 1H), 4.44 (s, 1H), 6.03 (d, J =
4.66 Hz, 1H), 6.65 (s, 1H), 7.35 (d, J = 8.79 Hz, 2H), 7.64 (d, J =
8.76 Hz, 2H), 7.7−7.83 (m, 2H), 8.69 (s, 1H), 8.8−8.92 (m, 2H).
HRMS-ESI (m/z): [M + H]+ calcd for C25H28N6O4S, 509.196 55;
found 509.196 32.
3-Cyclopropyl-1-[4-[4-[(3S)-3-methylmorpholin-4-yl]-6-(1-
methylsulfonylcyclopentyl)pyrimidin-2-yl]phenyl]urea (30). Yield
39%. 1H NMR (DMSO-d6) δ 0.40 (dd, J = 2.20, 3.75 Hz, 2H),
0.63 (dd, J = 1.97, 6.87 Hz, 2H), 1.21 (d, J = 6.77 Hz, 3H), 1.56 (d,
J = 7.00 Hz, 2H), 1.80 (dd, J = 3.02, 5.79 Hz, 2H), 2.36−2.46 (m,
2H), 2.55 (m, 1H), 2.67−2.72 (m, 1H), 2.75 (dt, J = 6.59, 6.59,
12.98 Hz, 1H), 2.88 (s, 3H), 3.19 (td, J = 3.86, 12.95, 13.04 Hz,
1H), 3.48 (td, J = 3.01, 11.84, 11.85 Hz, 1H), 3.63 (dd, J = 3.03,
11.48 Hz, 1H), 3.75 (d, J = 11.41 Hz, 1H), 3.96 (dd, J = 3.68, 11.36
Hz, 1H), 4.23 (d, J = 11.23 Hz, 1H), 4.54 (s, 1H), 6.42 (d, J = 2.60
Hz, 1H), 6.77 (s, 1H), 7.49 (d, J = 8.75 Hz, 2H), 8.22 (d, J = 8.76
Hz, 2H), 8.54 (s, 1H). 13C NMR (DMSO-d6) 6.31, 13.19, 22.34,
24.56, 31.67, 31.82, 37.04, 46.31, 66.03, 70.19, 77.21, 100.44, 117.11,
128.38, 130.45, 142.62, 155.68, 161.56, 161.96, 163.97. HRMS-ESI
(m/z): [M + H]+ calcd for C25H33N5O4S, 500.232 60; found
500.232 76.
1-[4-[4-(1-Cyclopropylsulfonylcyclopropyl)-6-[(3S)-3-methylmor-
pholin-4-yl]pyrimidin-2-yl]phenyl]-3-(2-hydroxyethyl)urea (38).
1
Yield 73%. H NMR (DMSO-d6) δ 0.88−0.95 (m, 2H), 1.02 (m,
3H), 1.22 (d, J = 6.78 Hz, 2H), 1.51−1.6 (m, 2H), 1.6−1.69 (m,
2H), 2.93−3.01 (m, 1H), 3.13−3.19 (m, 2H), 3.19 (d, J = 3.59 Hz,
1H), 3.45 (q, J = 5.51, 5.53, 5.53 Hz, 2H), 3.47 (d, J = 3.10 Hz,
1H), 3.62 (dd, J = 3.02, 11.48 Hz, 1H), 3.75 (d, J = 11.41 Hz, 1H),
3.96 (dd, J = 3.65, 11.37 Hz, 1H), 4.16 (s, 1H), 4.51 (s, 1H), 4.72
(t, J = 5.09, 5.09 Hz, 1H), 6.23 (s, 1H), 6.82 (s, 1H), 7.47 (d, J =
8.76 Hz, 2H), 8.19 (d, J = 8.77 Hz, 2H), 8.78 (s, 1H). HRMS-ESI
(m/z): [M + H]+ calcd for C24H31N5O5S, 502.211 87; found
502.211 98.
3-Cyclopropyl-1-[4-[4-[(3S)-3-methylmorpholin-4-yl]-6-(4-meth-
1
ylsulfonyloxan-4-yl)pyrimidin-2-yl]phenyl]urea (31). Yield 72%. H
NMR (CDCl3-d6) δ 0.71 (dd, J = 2.24, 3.51 Hz, 2H), 0.89 (dd, J =
1.95, 6.70 Hz, 2H), 1.35 (d, J = 6.79 Hz, 3H), 2.46−2.59 (m, 3H),
2.64 (dd, J = 2.37, 3.55 Hz, 1H), 2.71 (m, 4H), 3.28−3.5 (m, 3H),
3.56−3.67 (m, 1H), 3.73−3.88 (m, 2H), 3.97−4.13 (m, 3H), 4.20
(d, J = 12.79 Hz, 1H), 4.45 (s, 1H), 4.89 (s, 1H), 6.64 (s, 1H), 7.00
(s, 1H), 7.51 (d, J = 8.81 Hz, 2H), 8.31 (d, J = 8.79 Hz, 2H).
HRMS-ESI (m/z): [M + H]+ calcd for C25H33N5O5S, 516.227 52;
found 516.227 60.
3-(2-Hydroxyethyl)-1-[4-[4-[(3S)-3-methylmorpholin-4-yl]-6-(1-
methylsulfonylcyclobutyl)pyrimidin-2-yl]phenyl]urea (32). Yield
41%. 1H NMR (DMSO-d6) δ 1.22 (d, J = 6.77 Hz, 3H), 1.85−
1.93 (m, 1H), 2.01−2.1 (m, 1H), 2.74−2.83 (m, 2H), 2.85 (s, 3H),
2.89 (ddd, J = 5.05, 9.59, 16.90 Hz, 2H), 3.16 (d, J = 5.70 Hz, 1H),
3.17−3.23 (m, 1H), 3.45 (q, J = 5.50, 5.55, 5.55 Hz, 2H), 3.46−3.51
(m, 1H), 3.63 (dd, J = 3.09, 11.47 Hz, 1H), 3.75 (d, J = 11.41 Hz,
1H), 3.96 (dd, J = 3.67, 11.36 Hz, 1H), 4.22 (s, 1H), 4.55 (s, 1H),
4.71 (t, J = 5.13, 5.13 Hz, 1H), 6.23 (s, 1H), 6.68 (s, 1H), 7.47 (d, J
= 8.77 Hz, 1H), 8.20 (d, J = 8.81 Hz, 1H), 8.79 (s, 1H). HRMS-ESI
(m/z): [M + H]+ calcd for C23H31N5O5S, 490.211 87; found
490.211 88.
3-Cyclopropyl-1-[4-[4-[1-(3-hydroxypropylsulfonyl)cyclopropyl]-
6-[(3S)-3-methylmorpholin-4-yl]pyrimidin-2-yl]phenyl]urea (33).
Yield 59%. 1H NMR (DMSO-d6) δ 0.41 (dd, J = 2.14, 3.74 Hz,
2H), 0.63 (dd, J = 1.96, 6.86 Hz, 2H), 1.22 (d, J = 6.76 Hz, 3H),
1.5−1.58 (m, 2H), 1.59−1.68 (m, 2H), 1.86−1.97 (m, 2H), 2.55
(dq, J = 3.24, 3.25, 3.25, 6.94 Hz, 1H), 3.13−3.26 (m, 1H), 3.43−
3.55 (m, 5H), 3.62 (dd, J = 3.09, 11.49 Hz, 1H), 3.74 (d, J = 11.42
Hz, 1H), 3.95 (dd, J = 3.65, 11.37 Hz, 1H), 4.19 (s, 1H), 4.53 (s,
1H), 4.67 (t, J = 5.28, 5.28 Hz, 1H), 6.41 (d, J = 2.43 Hz, 1H), 6.75
(s, 1H), 7.49 (d, J = 8.76 Hz, 2H), 8.15−8.23 (m, J = 8.76 Hz, 2H),
8.52 (s, 1H). HRMS-ESI (m/z): [M + H]+ calcd for C25H33N5O5S,
516.227 52; found 516.227 54.
1-[4-[4-[1-(Benzenesulfonyl)cyclopropyl]-6-[(3S)-3-methylmor-
pholin-4-yl]pyrimidin-2-yl]phenyl]-3-(2-hydroxyethyl)urea (40).
Yield 88%. 1H NMR (DMSO-d6) δ 1.16 (d, J = 6.77 Hz, 3H),
1.57−1.68 (m, 2H), 1.88 (d, J = 2.73 Hz, 2H), 3.15 (dd, J = 7.11,
12.08 Hz, 3H), 3.42−3.47 (m, 4H), 3.59 (dd, J = 3.05, 11.51 Hz,
1H), 3.72 (d, J = 11.43 Hz, 1H), 3.94 (dd, J = 3.69, 11.39 Hz, 1H),
4.09 (s, 1H), 4.36 (s, 1H), 6.23 (s, 1H), 6.60 (s, 1H), 7.37 (d, J =
8.69 Hz, 2H), 7.58 (t, J = 7.84, 7.84 Hz, 2H), 7.70 (d, J = 7.45 Hz,
1H), 7.75−7.8 (m, 2H), 7.83 (d, J = 8.73 Hz, 2H), 8.77 (s, 1H). 13C
NMR (DMSO-d6) 13.14, 13.28, 21.29, 34.42, 37.38, 38.62, 41.74,
46.39, 47.10, 60.24, 65.95, 70.07, 101.48, 116.54, 128.43, 128.54,
129.07, 133.49, 139.04, 142.88, 154.93, 161.30, 161.71. HRMS-ESI
(m/z): [M + H]+ calcd for C27H31N5O5S, 538.211 87; found,
538.211 73.
1-[4-[4-[1-(Benzenesulfonyl)cyclopropyl]-6-[(3S)-3-methylmor-
pholin-4-yl]pyrimidin-2-yl]phenyl]-3-(3-hydroxypropyl)urea (41).
Yield 61%. 1H NMR (DMSO-d6) δ 1.15 (d, J = 6.77 Hz, 3H),
1.5−1.7 (m, 4H), 1.88 (m, 2H), 3.04−3.21 (m, 3H), 3.38−3.52 (m,
3H), 3.59 (dd, J = 3.07, 11.49 Hz, 1H), 3.72 (d, J = 11.41 Hz, 1H),
3.93 (dd, J = 3.69, 11.38 Hz, 1H), 4.08 (s, 1H), 4.36 (s, 1H), 4.45
(t, J = 5.13, 5.13 Hz, 1H), 6.16 (s, 1H), 6.60 (s, 1H), 7.3−7.41 (m,
2H), 7.57 (t, J = 7.84, 7.84 Hz, 2H), 7.66−7.73 (m, 1H), 7.73−7.88
(m, 4H), 8.65 (s, 1H). 13C NMR (DMSO-d6) 13.08, 13.20, 17.15,
28.92, 30.03, 32.75, 36.40, 38.54, 46.27, 47.27, 48.42, 58.51, 65.97,
70.11, 101.27, 116.55, 128.41, 129.04, 129.88, 133.42, 139.20, 142.76,
154.93, 160.50, 161.40, 161.92. HRMS-ESI (m/z): [M + H]+ calcd
for C28H33N5O5S, 552.227 52; found 552.227 36.
Compounds 19, 21, and 22 were prepared from intermediate J
according to the general procedure D. Compound 20 was prepared
from intermediate K according to the general procedure D.
General Proceedure D for Preparing Compounds. Inter-
mediate J or K (1 equiv) was treated with methanesulfonyl chloride
(1.5 equiv) and triethylamine (1.5 equiv) in DCM at 0 °C, then
allowed to come to ambient temperature and stirred until the
reaction was complete by TLC or LCMS. The reaction mixture was
concentrated in vacuo to yield the crude mesylate which was used
without further purification. DBU (2 equiv) was added to a solution
of the appropriate thiol (1.75 equiv) and the crude mesylate (1
equiv) in acetonitrile and the reaction stirred at ambient temperature
until complete by TLC or LCMS analysis. The reaction mixture was
concentrated in vacuo, and the crude thioether was used in the next
step either directly or following purification by HPLC. 3-
3-Cyclopropyl-1-[4-[4-(1-cyclopropylsulfonylcyclopropyl)-6-[(3S)-
3-methylmorpholin-4-yl]pyrimidin-2-yl]phenyl]urea (35). Yield
74%. 1H NMR (DMSO-d6) δ 0.33−0.47 (m, 2H), 0.63 (td, J =
4.85, 6.74, 6.82 Hz, 2H), 0.85−0.97 (m, 2H), 1.01 (dd, J = 2.13,
8.02 Hz, 2H), 1.22 (d, J = 6.77 Hz, 3H), 1.48−1.61 (m, 2H), 1.61−
1.71 (m, 2H), 2.55 (dd, J = 3.66, 6.89 Hz, 1H), 2.98 (ddd, J = 3.21,
4.66, 8.01 Hz, 1H), 3.12−3.25 (m, 1H), 3.47 (td, J = 3.06, 11.80,
11.83 Hz, 1H), 3.62 (dd, J = 3.08, 11.47 Hz, 1H), 3.75 (d, J = 11.41
Hz, 1H), 3.96 (dd, J = 3.67, 11.38 Hz, 1H), 4.17 (s, 1H), 4.51 (s,
1H), 6.40 (d, J = 2.53 Hz, 1H), 6.83 (s, 1H), 7.48 (d, J = 8.76 Hz,
2H), 8.20 (d, J = 8.76 Hz, 2H), 8.51 (s, 1H). HRMS-ESI (m/z): [M
+ H]+ calcd for C25H31N5O4S, 498.216 95; found, 498.217 10.
1-[4-[4-[1-(4-Fluorophenyl)sulfonylcyclopropyl]-6-[(3S)-3-methyl-
morpholin-4-yl]pyrimidin-2-yl]phenyl]-3-(2-hydroxyethyl)urea (36).
1
Yield 62%. H NMR (DMSO-d6) δ 1.18 (d, J = 6.71 Hz, 3H), 1.61
(d, J = 4.80 Hz, 2H), 1.82−1.98 (m, 2H), 3.17 (dd, J = 2.27, 5.40
Hz, 2H), 3.42−3.5 (m, 2H), 3.61 (dd, J = 2.84, 11.48 Hz, 2H), 3.74
Q
DOI: 10.1021/jm501778s
J. Med. Chem. XXXX, XXX, XXX−XXX

Journal of Medicinal Chemistry
Article
Chloroperbenzoic acid (75%, 2 equiv) and sodium permanganate
(2.5 equiv) were added to a solution of the thioether (1 equiv) in a
mixture of 1,4-dioxane and water. The reaction was stirred at
Hz, 1H), 4.51 (s, 2H), 6.83 (s, 1H), 7.63 (d, J = 8.72 Hz, 2H),
8.22−8.3 (m, 2H), 9.52 (s, 1H). HRMS-ESI (m/z): [M + H]+ calcd
for C21H27N5O3S2, 462.162 81; found 462.162 63.
3-(2-Hydroxyethyl)-1-[4-[4-[(3S)-3-methylmorpholin-4-yl]-6-(2-
methylsulfonylpropan-2-yl)pyrimidin-2-yl]phenyl]thiourea (34).
Yield 41%. 1H NMR (DMSO-d6) δ 1.24 (d, J = 6.72 Hz, 3H),
1.78 (d, J = 2.25 Hz, 6H), 3.04 (s, 3H), 3.18−3.24 (m, 1H), 3.50 (d,
J = 2.85 Hz, 1H), 3.57 (m, 4H), 3.65 (dd, J = 2.93, 11.47 Hz, 1H),
3.77 (d, J = 11.45 Hz, 1H), 3.98 (dd, J = 3.36, 11.25 Hz, 1H), 4.24
(d, J = 13.50 Hz, 1H), 4.63 (s, 1H), 4.81 (s, 1H), 6.78 (s, 1H), 7.63
(d, J = 8.75 Hz, 2H), 7.87 (s, 1H), 8.30 (d, J = 8.74 Hz, 2H), 9.81
(s, 1H). HRMS-ESI (m/z): [M + H]+ calcd for C22H31N5O4S2,
494.189 02; found 494.188 87.
3-(2-Hydroxyethyl)-1-[4-[4-[(3S)-3-methylmorpholin-4-yl]-6-(1-
methylsulfonylcyclopropyl)pyrimidin-2-yl]phenyl]thiourea (39).
Yield 81%.1H NMR (DMSO-d6) δ 1.23 (d, J = 6.77 Hz, 3H),
1.55 (d, J = 2.29 Hz, 2H), 1.66 (t, J = 3.33, 3.33 Hz, 2H), 3.20 (td, J
= 3.86, 12.92, 13.04 Hz, 1H), 3.29 (s. 3H), 3.47 (td, J = 3.07, 11.79,
11.82 Hz, 1H), 3.52−3.6 (m, 4H), 3.62 (dd, J = 3.09, 11.49 Hz,
1H), 3.75 (d, J = 11.43 Hz, 1H), 3.96 (dd, J = 3.67, 11.37 Hz, 1H),
4.20 (s, 1H), 4.57 (s, 1H), 4.79 (s, 1H), 6.79 (s, 1H), 7.62 (d, J =
8.70 Hz, 2H), 7.85 (s, 1H), 8.24 (d, J = 8.70 Hz, 2H), 9.79 (s, 1H).
HRMS-ESI (m/z): [M + H]+ calcd for C22H29N5O4S2, 492.173 37;
found 492.173 31.
1-[4-[4-(1-Cyclopropylsulfonylcyclopropyl)-6-[(3S)-3-methylmor-
pholin-4-yl]pyrimidin-2-yl]phenyl]-3-(2-hydroxyethyl)thiourea (42
(AZD3147)). Yield 77%. 1H NMR (DMSO-d6) δ 0.92 (dd, J =
1.63, 3.29 Hz, 2H), 1.02 (dd, J = 2.16, 8.04 Hz, 2H), 1.23 (d, J =
6.77 Hz, 3H), 1.57 (dd, J = 1.12, 6.00 Hz, 2H), 1.64 (dd, J = 2.73,
6.38 Hz, 2H), 2.92−3.01 (m, 1H), 3.21 (td, J = 3.89, 12.88, 13.00
Hz, 1H), 3.48 (td, J = 3.07, 11.83, 11.83 Hz, 1H), 3.51−3.59 (m,
4H), 3.63 (dd, J = 3.09, 11.50 Hz, 1H), 3.75 (d, J = 11.43 Hz, 1H),
3.96 (dd, J = 3.70, 11.38 Hz, 1H), 4.17 (s, 1H), 4.54 (s, 1H), 4.79
(s, 1H), 6.87 (s, 1H), 7.61 (d, J = 8.71 Hz, 2H), 7.83 (s, 1H), 8.26
(d, J = 8.68 Hz, 2H), 9.78 (s, 1H). 13C NMR (DMSO-d6) 4.44, 4.51,
12.13, 12.15, 29.29, 38.70, 45.96, 46.40, 59.10, 65.99, 70.17, 101.44,
121.53, 128.12, 132.84, 141.60, 161.84, 161.65, 161.51, 180.24.
HRMS-ESI (m/z): [M + H]+ calcd for C24H31N5O4S2, 518.189 02;
found 518.188 96.
ambient temperature for up to 18
h with additional 3-
chloroperbenzoic acid and sodium permanganate added if required
to push the reaction to completion by TLC or LCMS analysis. The
reaction mixture was purified by ion exchange cartridge (SCX) with
the desired material eluted with 7 N ammonia in methanol. The
crude material was further purified by HPLC.
1-[4-[4-(Cyclohexylsulfonylmethyl)-6-[(3S)-3-methylmorpholin-4-
yl]pyrimidin-2-yl]phenyl]-3-methylurea (19). Yield 43%. 1H NMR
(DMSO-d6) δ 1.23 (d, J = 6.74 Hz, 5H), 1.43 (d, J = 12.48 Hz, 2H),
1.66 (d, J = 12.46 Hz, 1H), 1.87 (d, J = 12.98 Hz, 2H), 2.23 (s, 2H),
2.65 (d, J = 4.61 Hz, 3H), 3.15−3.24 (m, 1H), 3.34 (d, J = 8.68 Hz,
1H), 3.48 (td, J = 2.95, 11.84, 11.84 Hz, 1H), 3.63 (dd, J = 2.98,
11.50 Hz, 1H), 3.76 (d, J = 11.42 Hz, 1H), 3.97 (dd, J = 3.54, 11.35
Hz, 1H), 4.15 (s, 1H), 4.41 (s, 1H), 6.07 (d, J = 4.65 Hz, 1H), 6.75
(s, 1H), 7.49 (d, J = 8.76 Hz, 2H), 8.18 (d, J = 8.75 Hz, 2H), 8.73
(s, 1H). HRMS-ESI (m/z): [M + H]+ calcd for C24H33N5O4S,
488.232 60; found, 488.232 51.
3-Cyclopropyl-1-[4-[4-(3-hydroxypropylsulfonylmethyl)-6-[(3S)-3-
methylmorpholin-4-yl]pyrimidin-2-yl]phenyl]urea (20). Yield 17%.
1H NMR (DMSO-d6) δ 0.42 (dd, J = 2.13, 3.74 Hz, 2H), 0.65 (dd, J
= 2.02, 6.86 Hz, 2H), 1.24 (d, J = 6.72 Hz, 3H), 1.96 (dd, J = 5.74,
10.22 Hz, 2H), 2.53−2.59 (m, 1H), 3.22 (d, J = 3.47 Hz, 1H),
3.34−3.46 (m, 2H), 3.45−3.61 (m, 3H), 3.65 (dd, J = 2.94, 11.47
Hz, 1H), 3.78 (d, J = 11.48 Hz, 1H), 3.99 (dd, J = 3.37, 11.39 Hz,
1H), 4.16 (s, 1H), 4.46 (s, 3H), 4.72 (t, J = 5.23, 5.23 Hz, 1H), 6.43
(d, J = 2.76 Hz, 1H), 6.78 (s, 1H), 7.50 (d, J = 8.84 Hz, 2H), 8.21
(d, J = 8.82 Hz, 2H), 8.53 (s, 1H). HRMS-ESI (m/z): [M + H]+
calcd for C23H31N5O5S, 490.211 87; found 490.211 85.
1-[4-[4-(Benzenesulfonylmethyl)-6-[(3S)-3-methylmorpholin-4-
yl]pyrimidin-2-yl]phenyl]-3-methylurea (21). Yield 24%. 1H NMR
(DMSO-d6) δ 1.21 (d, J = 6.73 Hz, 3H), 2.67 (d, J = 4.42 Hz, 3H),
3.07−3.26 (m, 1H), 3.49 (td, J = 2.91, 11.77, 11.83 Hz, 1H), 3.64
(dd, J = 2.97, 11.51 Hz, 1H), 3.77 (d, J = 11.45 Hz, 1H), 3.98 (dd, J
= 3.42, 11.35 Hz, 1H), 4.10 (d, J = 13.03 Hz, 1H), 4.38 (s, 1H),
4.71 (s, 2H), 6.06 (d, J = 4.64 Hz, 1H), 6.61 (s, 1H), 7.39 (d, J =
8.80 Hz, 2H), 7.63 (t, J = 7.72, 7.72 Hz, 2H), 7.73−7.78 (m, 1H),
7.8−7.85 (m, 4H), 8.71 (s, 1H). HRMS-ESI (m/z): [M + H]+ calcd
for C24H27N5O4S, 482.185 65; found, 482.185 55.
3-Cyclopropyl-1-[4-[4-[1-(3-hydroxypropylsulfonyl)cyclopropyl]-
6-[(3S)-3-methylmorpholin-4-yl]pyrimidin-2-yl]phenyl]thiourea
1
(43). Yield 98%. H NMR (DMSO-d6) δ 0.59 (s, 2H), 0.75 (d, J =
3-Methyl-1-[4-[4-[(3S)-3-methylmorpholin-4-yl]-6-(pyridin-4-
ylsulfonylmethyl)pyrimidin-2-yl]phenyl]urea (22). Yield 15%. 1H
NMR (DMSO-d6) δ 1.19 (d, J = 6.78 Hz, 3H), 2.64 (d, J = 4.58 Hz,
3H), 3.17 (td, J = 3.86, 12.89, 12.99 Hz, 1H), 3.46 (td, J = 3.04,
11.83, 11.84 Hz, 1H), 3.62 (dd, J = 3.08, 11.48 Hz, 1H), 3.75 (d, J =
11.41 Hz, 1H), 3.95 (dd, J = 3.67, 11.37 Hz, 1H), 4.10 (s, 1H), 4.37
(s, 1H), 4.85 (d, J = 1.48 Hz, 2H), 6.04 (d, J = 4.62 Hz, 1H), 6.69
(s, 1H), 7.35 (d, J = 8.78 Hz, 2H), 7.56−7.71 (d, J = 8.78 Hz, 2H),
7.72−7.86 (m, 2H), 8.70 (s, 1H), 8.84−8.96 (m, 2H). Mass
calculated for C23H26N6O4S 482.56, found 483.45 (ESI, M + H).
Compounds 23, 34, 39, and 42 were prepared from intermediates
E, O, Q, and AH, respectively, according to the general procedure E.
Compounds 43 and 44 were prepared from intermediate Z
according to the general procedure E.
General Proceedure E for Preparing Compounds. A solution
of di(imidazol-1-yl)methanethione (1.3 equiv) in DCM was added to
a solution of the appropriate intermediate (1 equiv) in either DCM
or a mixture of DCM and THF. The reaction was stirred at ambient
temperature for 3 h and then the appropriate amine (5 equiv) added
followed by triethylamine (1 equiv). The reaction was stirred at
ambient temperature for 1 h or until complete by TLC or LCMS
analysis. The reaction mixture was concentrated in vacuo and the
residue purified by HPLC.
5.36 Hz, 2H), 1.23 (d, J = 6.79 Hz, 3H), 1.53−1.59 (m, 2H), 1.6−
1.67 (m, 2H), 1.88−1.97 (m, 2H), 2.91 (s, 1H), 3.16−3.25 (m, 1H),
3.49 (m, 5H), 3.62 (dd, J = 3.07, 11.49 Hz, 1H), 3.75 (d, J = 11.43
Hz, 1H), 3.96 (dd, J = 3.65, 11.37 Hz, 1H), 4.20 (s, 1H), 4.56 (s,
1H), 4.67 (t, J = 5.26, 5.26 Hz, 1H), 6.80 (s, 1H), 7.60 (d, J = 8.62
Hz, 2H), 8.13 (s, 1H), 8.25 (d, J = 8.66 Hz, 2H), 9.48 (s, 1H). 13C
NMR (DMSO-d6) 6.77, 11.63, 11.65, 13.29, 25.03, 38.69, 45.00,
46.27, 49.60, 59.03, 65.99, 70.16, 101.12, 127.85, 132.96, 141.90,
161.52, 161.79, 161.96, 181.44. HRMS-ESI (m/z): [M + H]+ calcd
for C25H33N5O4S2, 532.204 67; found 532.204 77.
3-(2-Hydroxyethyl)-1-[4-[4-[1-(3-hydroxypropylsulfonyl)-
cyclopropyl]-6-[(3S)-3-methylmorpholin-4-yl]pyrimidin-2-yl]-
1
phenyl]thiourea (44). Yield >99%. H NMR (DMSO-d6) δ 1.22 (d,
J = 6.78 Hz, 3H), 1.55 (d, J = 5.06 Hz, 2H), 1.58−1.69 (m, 2H),
1.85−1.98 (m, 2H), 3.20 (td, J = 3.82, 12.92, 13.03 Hz, 1H), 3.45−
3.52 (m, 5H), 3.56 (s, 4H), 3.62 (dd, J = 3.01, 11.49 Hz, 1H), 3.75
(d, J = 11.42 Hz, 1H), 3.96 (dd, J = 3.58, 11.35 Hz, 1H), 4.19 (s,
1H), 4.56 (s, 1H), 4.67 (t, J = 5.25, 5.25 Hz, 1H), 4.79 (s, 1H), 6.80
(s, 1H), 7.61 (d, J = 8.66 Hz, 2H), 7.84 (s, 1H), 8.25 (d, J = 8.62
Hz, 2H), 9.79 (s, 1H). 13C NMR (DMSO-d6) 11.65, 13.30, 25.03,
45.00, 46.27, 46.40, 49.58, 59.02, 65.99, 70.16, 101.12, 121.55,
128.13, 132.73, 141.68, 161.52, 161.78, 161.93, 180.26. HRMS-ESI
(m/z): [M + H]+ calcd for C24H33N5O5S2, 536.199 59; found
536.199 58.
The prepartion of the intermediates used in the above procedures
are described below.
Methyl 2-Chloro-6-[(3S)-3-methylmorpholin-4-yl]pyrimidine-4-
carboxylate (Intermediate A). A solution of (3S)-3-methylmorpho-
line (2.49 g) and triethylamine (3.70 mL) in DCM (10 mL) was
3-Cyclopropyl-1-[4-[4-[(3S)-3-methylmorpholin-4-yl]-6-
(methylsulfonylmethyl)pyrimidin-2-yl]phenyl]thiourea (23). Yield
47%. 1H NMR (DMSO-d6) δ 0.60 (s, 2H), 0.71−0.86 (m, 2H),
1.26 (d, J = 6.73 Hz, 3H), 2.91 (d, J = 12.23 Hz, 1H), 3.25 (s, 1H),
3.48−3.6 (m, 1H), 3.66 (dd, J = 2.98, 11.53 Hz, 1H), 3.78 (d, J =
11.46 Hz, 2H), 3.99 (dd, J = 3.39, 11.37 Hz, 2H), 4.18 (d, J = 11.30
R
DOI: 10.1021/jm501778s
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Journal of Medicinal Chemistry
Article
added dropwise to a solution of methyl 2,6-dichloropyrimidine-4-
carboxylate (5 g) in DCM (120 mL) over 10 min and the reaction
stirred at ambient temperature for 1 h. The reaction was
concentrated in vacuo, the residue dissolved in DCM and washed
with water. The organics were dried (MgSO4), filtered, and
evaporated and the residue chromatographed on silica, eluting with
2.5% methanol in DCM, to give the desired material as a white solid
3-[[2-Chloro-6-[(3S)-3-methylmorpholin-4-yl]pyrimidin-4-yl]-
methylsulfonyl]propan-1-ol (Intermediate X). A solution of
intermediate C (12.4 g, 35.07 mmol) in DCM (50 mL) was
added to a stirred solution of 3-mercapto-1-propanol (3.64 mL,
42.08 mmol) and DIPEA (9.77 mL, 56.11 mmol) in DCM (100
mL) over a period of 40 min under a nitrogen atmosphere. The
resulting solution was stirred at ambient temperature for 18 h and
then washed sequentially with a saturated aqueous solution of
sodium hydrogen carbonate (2 × 50 mL) and saturated brine (50
mL). The organic layer was dried (MgSO4), filtered, and evaporated
to afford crude product as a dark brown oil. The crude product was
purified by flash silica chromatography, eluting with 0−75% ethyl
acetate in DCM, to give 3-[[2-chloro-6-[(3S)-3-methylmorpholin-4-
yl]pyrimidin-4-yl]methylsulfanyl]propan-1-ol as a yellow gum (5.86
1
(3.15 g, yield 48%). H NMR (DMSO-d6) δ 1.22−1.24 (3H, m),
3.25 (1H, d), 3.41−3.48 (1H, m), 3.57−3.61 (1H, m), 3.71 (1H, d),
3.87 (3H, s), 3.91−3.95 (1H, m), 4.25 (1H, s), 4.45 (1H, s), 7.29
(1H, s). ESI (m/z): [M + H]+ = 272.
[2-Chloro-6-[(3S)-3-methylmorpholin-4-yl]pyrimidin-4-yl]-
methanol (Intermediate B). Intermediate A (3.15 g) was dissolved
in dry THF (20 mL) and cooled to 0 °C under nitrogen. A solution
of lithium borohydride (2.0 M in THF, 6.09 mL) was added
dropwise and the solution allowed to warm to rt and stirred for 1 h.
The reaction was quenched with water (20 mL), then evaporated to
dryness, the residue dissolved in ethyl acetate (150 mL) and washed
with water (150 mL) followed by brine (50 mL). The organics were
evaporated to dryness to give the desired material as a white solid
1
g, yield 53%). H NMR (CDCl3) δ 1.32 (3H, d), 1.84−1.90 (2H,
m), 1.94 (1H, s), 2.69 (2H, t), 3.24−3.32 (1H, m), 3.51−3.58 (1H,
m), 3.61 (2H, s), 3.67−3.71 (1H, m), 3.73−3.80 (3H, m), 3.98−4.04
(2H, m), 4.28−4.34 (1H, m), 6.45 (1H, s). ESI (m/z): [M + H]+ =
318. 3-Chlorobenzoperoxoic acid (4.00 g, 23.16 mmol) was added to
3-[[2-chloro-6-[(3S)-3-methylmorpholin-4-yl]pyrimidin-4-yl]-
methylsulfanyl]propan-1-ol (3.68 g, 11.58 mmol) in DCM (100 mL)
and the resulting solution stirred at ambient temperature for 3 h. A
further portion of 3-chlorobenzoperoxoic acid (2.00 g, 11.58 mmol)
was added and the resulting solution stirred at ambient temperature
for 1 h. The reaction mixture was washed sequentially with 10%
aqueous sodium metabisulfite solution (2 × 100 mL), a saturated
aqueous solution of sodium hydrogen carbonate (100 mL), and
saturated brine (100 mL). The organic layer was dried (MgSO4),
filtered, and evaporated to give the desired material as a gum (4.05 g,
1
(2.44 g, yield 66%). H NMR (DMSO-d6) δ 1.20−1.21 (3H, m),
3.18−3.22 (1H, m), 3.40−3.47 (1H, m), 3.56−3.60 (1H, m), 3.71
(1H, d), 3.91−3.94 (1H, m), 3.98 (1H, d), 4.35 (3H, d), 5.51 (1H,
t), 6.74 (1H, s). ESI (m/z): [M + H]+ = 244.
2-Chloro-4-(iodomethyl)-6-[(3S)-3-methylmorpholin-4-yl]-
pyrimidine (Intermediate C). Methanesulfonyl chloride (0.245 mL,
3.14 mmol) was added dropwise over a period of 5 min to a solution
of triethylamine (0.875 mL, 6.28 mmol) and intermediate B (510
mg, 2.09 mmol) in DCM (30 mL) at 0 °C under nitrogen. The
resulting solution was stirred at ambient temperature for 45 min and
then diluted with water (20 mL). The organic layer was dried
(MgSO4) and filtered. Sodium iodide (1569 mg, 10.46 mmol) was
added and the reaction heated at 50 °C for 20 h. The reaction
mixture was filtered and evaporated to afford the desired material
which was used crude in further reactions (761 mg, yield ∼100%).
1H NMR (DMSO-d6) δ 1.19−1.25 (3H, m), 3.18−3.22 (1H, m),
1
yield 100%). H NMR (CDCl3) δ 1.34 (3H, d), 2.12−2.18 (2H, m),
3.27 (2H, t), 3.31−3.35 (1H, m), 3.51−3.57 (1H, m), 3.67−3.70
(1H, m), 3.77−3.82 (3H, m), 3.99−4.03 (1H, m), 4.18 (2H, s),
4.26−4.37 (1H, m), 6.51 (1H, s). ESI (m/z): [M + H]+ = 350.
2-Chloro-4-[(4-fluorophenyl)sulfonylmethyl]-6-[(3S)-3-methyl-
morpholin-4-yl]pyrimidine (Intermediate AB). 4-Fluorobenzenesul-
finic acid sodium salt (6.70 g, 36.77 mmol) was added to
intermediate C (10.00 g, 28.28 mmol) in acetonitrile (250 mL)
under nitrogen and the resulting solution stirred at 80 °C for 20 h.
The reaction was cooled, the solvent was removed in vacuo and the
residue partitioned between DCM and water. The organic layer was
dried over MgSO4 and concentrated in vacuo to give crude material
which was purified by flash silica chromatography, elution gradient
0−30% ethyl acetate in DCM, to give the desired material as a cream
solid (8.32 g, yield 76%). 1H NMR (DMSO-d6) δ 1.17 (3H, d),
3.13−3.20 (1H, m), 3.27−3.28 (1H, m), 3.39−3.46 (1H, m), 3.57
(1H, dd), 3.72 (1H, d), 3.93 (1H, dd), 4.17 (1H, s), 4.65 (2H, s),
6.71 (1H, s), 7.48 (2H, t), 7.83−7.87 (2H, m). ESI (m/z): [M +
H]+ = 386.
3.40−3.47 (1H, m), 3.57−3.60 (1H, m), 3.71 (1H, d), 3.90−3.94
(1H, m), 3.96−3.98 (1H, m), 4.28−4.32 (3H, m), 6.94 (1H, s). ESI
(m/z): [M + H]+ = 354.
2 - C h l o r o - 4 - [ ( 3 S ) - 3 - m e t h y l m o r p h o l i n - 4 - y l ] - 6 -
(methylsulfonylmethyl)pyrimidine (Intermediate D). Methanesul-
finic acid, sodium salt (11.75 g, 115.11 mmol) was added in one
portion to intermediate C (37 g, 104.64 mmol) in acetonitrile (900
mL) and the resulting solution stirred at 85 °C for 24 h. The
mixture was concentrated in vacuo and the residue dissolved in
DCM. The organic layers were combined and washed with water (3
× 100 mL), dried over MgSO4, filtered, and the solvent was removed
by evaporation to give the crude product as a dark brown oil, which
solidifed (36 g). The crude solid was purified by flash silica
chromatography, elution gradient 0−30% ethyl acetate in DCM, to
give the desired material as a cream solid (22 g, yield 69%). 1H
NMR (DMSO-d6) δ1.21−1.23 (m, 3H), 3.11 (s, 3H), 3.19−3.26 (m,
1H), 3.42−3.49 (m, 1H), 3.58−3.62 (1H, m), 3.73 (d, 1H), 3.92−
3.96 (m, 2H), 4.27−4.31 (m, 1H), 4.45 (s, 2H), 6.92 (s, 1H). ESI
(m/z): [M + H]+ = 306.
2-Chloro-4-[(3S)-3-methylmorpholin-4-yl]-6-(pyridin-4-
ylsulfonylmethyl)pyrimidine (Intermediate AA). Intermediate C
(5.00 g, 14.14 mmol) was added portionwise to 4-mercaptopyridine
(1.572 g, 14.14 mmol) and DIPEA (4.93 mL, 28.28 mmol) in
acetonitrile (120 mL) over a period of 10 min. The resulting
solution was stirred at ambient temperature for 3 h before being
concentrated in vacuo and the residue partitioned between DCM
and water. The organics were dried over MgSO4, concentrated in
vacuo and the residue was purified by flash silica chromatography,
elution gradient 0−4% methanol in DCM, to yield 2-chloro-4-[(3S)-
3-methylmorpholin-4-yl]-6-(pyridin-4-ylsulfanylmethyl)pyrimidine as
2-Chloro-4-(cyclopropylsulfonylmethyl)-6-[(3S)-3-methylmor-
pholin-4-yl]pyrimidine (Intermediate L). Cyclopropanesulfinic acid,
sodium salt (381 mg, 2.97 mmol) was added in one portion to
intermediate C (700 mg, 1.98 mmol) in acetonitrile (20 mL) and the
resulting suspension stirred at 90 °C for 3 h. The reaction mixture
was evaporated to dryness and redissolved in DCM (50 mL) and
washed with water (50 mL). The organic layer was dried (MgSO4),
filtered, and evaporated to afford crude product. The crude product
was purified by flash silica chromatography, elution gradient 0−40%
ethyl acetate in DCM, to give the desired material as a white solid
1
a yellow solid (4.41 g, yield 93%). H NMR (DMSO-d6) δ 1.14−
1.16 (3H, d), 3.11−3.18 (1H, td), 3.37−3.44 (1H, td), 3.53−3.57
(1H, dd), 3.64−3.67 (1H, d), 3.86−3.90 (2H, dd), 4.01 (2H, s), 4.14
(1H, bs), 6.43 (1H, s), 7.04−7.06 (2H, d), 8.29−8.30 (2H, d). ESI
(m/z): [M + H]+ = 337. A 30% aqueous solution of hydrogen
peroxide (8.87 mL, 286.78 mmol) was added dropwise to a stirred
solution of 2-chloro-4-[(3S)-3-methylmorpholin-4-yl]-6-(pyridin-4-
ylsulfanylmethyl)pyrimidine (4.83 g, 14.34 mmol) and sodium
tungstate dihydrate (0.095 g, 0.29 mmol) in water (3.5 mL) and 2
N sulfuric acid (0.358 mL) in dioxane (48 mL) and methanol (2
mL). The resulting solution was stirred at 55 °C for 4 h and then
1
(458 mg, yield 70%). H NMR (DMSO-d6) δ 0.95−0.98 (2H, m),
1.02−1.06 (2H, m), 1.18−1.23 (3H, m), 2.77−2.83 (1H, m), 3.19−
3.25 (1H, m), 3.42−3.49 (1H, m), 3.58−3.62 (1H, m), 3.73 (1H, d),
3.92−3.96 (2H, m), 4.30 (1H, s), 4.48 (2H, s), 6.92 (1H, s). ESI
(m/z): [M + H]+ = 332.
S
DOI: 10.1021/jm501778s
J. Med. Chem. XXXX, XXX, XXX−XXX

Journal of Medicinal Chemistry
Article
diluted with water and cooled. A 10% aqueous solution of sodium
metabisulfite was added and the mixture extracted with DCM, and
the organics were dried over MgSO4 and filtered to obtain the
3.91 (2H, dd), 4.25 (1H, s), 6.70 (1H, s), 7.45 (2H, t), 7.79−7.84
(2H, m). ESI (m/z): [M + H]+ = 412.
2-Chloro-4-[(3S)-3-methylmorpholin-4-yl]-6-(1-pyridin-4-
1
1
ylsulfonylcyclopropyl)pyrimidine (Intermediate AF). Yield 82%. H
desired material as a solid (4.47 g, 85%). H NMR (DMSO-d6) δ
NMR (DMSO-d6) δ 1.15−1.16 (3H, d), 1.61−1.65 (2H, m), 1.90−
1.93 (2H, m), 3.11−3.19 (1H, td), 3.37−3.44 (1H, td), 3.53−3.57
(1H, dd), 3.68−3.71 (1H, d), 3.89−3.96 (1H, dd), 3.96 (1H, bs),
4.28 (1H, bs), 6.75 (1H, s), 7.74−7.75 (2H, dd), 8.88−8.90 (2H,
dd). ESI (m/z): [M + H]+ = 395.
1.17−1.19 (3H, d), 3.14−3.22 (1H, td), 3.40−3.47 (1H, td), 3.56−
3.60 (1H, dd), 3.71−3.74 (1H, d), 3.90 (1H, bs), 3.91−3.95 (1H,
dd), 4.20 (1H, bs), 4.79 (2H, s), 6.79 (1H, s), 7.77−7.79 (2H, q),
8.92−8.93 (2H, q). ESI (m/z): [M + H]+ = 369.
4-(Benzenesulfonylmethyl)-2-chloro-6-[(3S)-3-methylmorpholin-
4-yl]pyrimidine (Intermediate AC). Benzenesulfinic acid, sodium salt
(4.22 g, 25.74 mmol) was added to intermediate C (7.0 g, 19.80
mmol) in acetonitrile (200 mL) and the resulting mixture stirred
under a nitrogen atmosphere at 80 °C for 20 h. The reaction was
cooled, and the solvent was removed. DCM was added, and the
solution was washed with water. The organics were dried (MgSO4),
filtered, and the solvent was removed. The crude product was
purified by flash silica chromatography, elution gradient 0−30% ethyl
acetate in DCM, to give the desired material as a cream solid (6.21
4-[1-(Benzenesulfonyl)cyclopropyl]-2-chloro-6-[(3S)-3-methyl-
1
morpholin-4-yl]pyrimidine (Intermediate AG). Yield 93%. H NMR
(DMSO-d6) δ 1.13−1.15 (3H, d), 1.55−1.57 (2H, m), 1.83−1.86
(2H, m), 3.09−3.16 (1H, td), 3.36−3.43 (1H, td), 3.52−3.56 (1H,
dd), 3.68−3.71 (1H, d), 3.86−3.93 (2H, m), 4.20 (1H, bs), 6.67
(1H, s), 7.60−7.63 (2H, m), 7.72−7.77 (3H, m). ESI (m/z): [M +
H]+ = 394.
3-[1-[2-Chloro-6-[(3S)-3-methylmorpholin-4-yl]pyrimidin-4-yl]-
cyclopropyl]sulfonylpropan-1-ol (Intermediate Y). To a solution of
intermediate X (5.04 g, 14.41 mmol) in DMF (25 mL) was added a
solution of chlorotrisopropylsilane (3.70 mL, 17.29 mmol) and
imidazole (2.354 g, 34.58 mmol) in DMF (25 mL) over a period of
5 min under a nitrogen atmosphere. The resulting solution was
stirred at ambient temperature for 18 h. The reaction mixture was
evaporated to dryness, the residue dissolved in DCM (200 mL) and
then washed sequentially with water (100 mL) and saturated brine
(100 mL). The organic layer was dried (MgSO4), filtered, and
evaporated to give 3-[[2-chloro-6-[(3S)-3-methylmorpholin-4-yl]-
pyrimidin-4-yl]methylsulfonyl]propoxytri(propan-2-yl)silane as an
oil (7.29 g, yield 100%). 1H NMR (CDCl3) δ 0.99−1.07 (21H,
m), 1.33 (3H, d), 2.06−2.13 (2H, m), 3.20−3.24 (2H, m), 3.26−
3.34 (1H, m), 3.50−3.57 (1H, m), 3.66−3.70 (1H, m), 3.77−3.83
(3H, m), 3.99−4.03 (2H, m), 4.16 (2H, s), 4.25−4.37 (1H, m), 6.54
(1H, s). ESI (m/z): [M + H]+ = 506. 1,2-Dibromoethane (1.723
mL, 20 mmol) was added to 3-[[2-chloro-6-[(3S)-3-methylmorpho-
lin-4-yl]pyrimidin-4-yl]methylsulfonyl]propoxytri(propan-2-yl)silane
(5.0 g, 9.88 mmol), 40% sodium hydroxide solution (10 mL, 98.78
mmol), and tetrabutylammonium bromide (0.645 g, 2 mmol) in
toluene (50 mL). The resulting mixture was stirred at 60 °C for 4 h.
The reaction mixture was evaporated to dryness and redissolved in
ethyl acetate (100 mL) and washed sequentially with water (100
mL) and saturated brine (100 mL). The organic layer was dried
(MgSO4), filtered, and evaporated to afford crude product. The
crude product was purified by flash silica chromatography, eluting
with 0−20% ethyl acetate in DCM, to give 3-[1-[2-chloro-6-[(3S)-3-
methylmorpholin-4-yl]pyrimidin-4-yl]cyclopropyl]sulfonylpropoxytri-
1
g, yield 85%). H NMR (DMSO-d6) δ 1.15−1.16 (3H, d), 3.11−3.18
(1H, td), 3.38−3.45 (1H, td), 3.55−3.58 (1H, dd), 3.70−3.73 (1H,
d), 3.85−3.94 (2H, m), 4.15 (1H, bs), 4.64 (2H, s), 6.67 (1H, s),
7.63−7.66 (2H, m), 7.74−7.80 (3H, m). ESI (m/z): [M + H]+
368.
=
2-Chloro-4-[(3S)-3-methylmorpholin-4-yl]-6-(2-methylsulfonyl-
propan-2-yl)pyrimidine (Intermediate N). Sodium tert-butoxide
(278 mg, 2.89 mmol) was added to intermediate D (883 mg, 2.89
mmol) in DMF (25 mL) at 0 °C under nitrogen. Iodomethane
(0.180 mL, 2.89 mmol) was added, and the resulting solution was
stirred at 0 °C for 15 min. Further sodium tert-butoxide (278 mg,
2.89 mmol) was added followed by iodomethane (0.180 mL, 2.89
mmol), and the resulting solution was stirred at 0 °C for 1 h. The
reaction was diluted with DCM (100 mL) and washed with water
(100 mL) and brine (100 mL). The organic layer was dried
(MgSO4), filtered and the solvent evaporated to a gum which slowly
crystallized.The crude product was purified by flash silica
chromatography, eluting with 0−5% methanol in DCM, to give
1
the desired material as a white solid (691 mg, yield 72%). H NMR
(DMSO-d6) δ 1.26 (3H, d), 1.72 (6H, s), 2.87 (3H, s), 3.19−3.27
(1H, m), 3.44−3.51 (1H, m), 3.60−3.63 (1H, m), 3.72 (1H, d),
3.92−3.96 (2H, m), 4.23−4.32 (1H, m), 6.53 (1H, s). ESI (m/z):
[M + H]+ = 334.
Intermediates P, AD, AE, AF, and AG were prepared from
intermediates D, L, AB, AA, and AC respectively, according to the
general procedure F.
General Procedure F. A mixture of the appropriate intermediate
(1 equiv), tetrabutylammonium bromide (0.1−0.2 equiv) and 1,2-
dibromoethane (2−5 equiv) in an aqueous solution of sodium
hydroxide (large excess) and toluene was heated at 60−70 °C for 4
h or until complete by either TLC or LCMS analysis. The reaction
mixture was either washed directly with water or partitioned between
ethyl acetate or DCM and water. The organics were dried over
MgSO4, concentrated in vacuo, and purified by either flash silica
chromatography or trituration with diethyl ether.
1
(propan-2-yl)silane as a colorless gum (2.86 g, yield 54%). H NMR
(CDCl3) δ 1.00−1.05 (21H, m), 1.32 (3H, d), 1.49−1.52 (2H, m),
1.78−1.81 (2H, m), 2.02−2.09 (2H, m), 3.21−3.32 (3H, m), 3.50−
3.56 (1H, m), 3.65−3.69 (1H, m), 3.77−3.80 (3H, m), 3.98−4.02
(2H, m), 4.28−4.36 (1H, m), 6.90 (1H, s). ESI (m/z): [M + H]+ =
532. Tetrabutylammonium fluoride (1 M in THF, 31 mL, 31 mmol)
was added to 3-[1-[2-chloro-6-[(3S)-3-methylmorpholin-4-yl]-
pyrimidin-4-yl]cyclopropyl]sulfonylpropoxytri(propan-2-yl)silane
(3.28 g, 6.16 mmol) in THF (30 mL). The resulting solution was
stirred at ambient temperature for 1 h and then concentrated in
vacuo and diluted with saturated ammonium chloride (10 mL) and
water. The mixture was extracted twice with ethyl acetate, and the
combined organic extracts were washed with brine, dried (MgSO4),
filtered, and evaporated to give the desired material as a gum (2.33 g,
2 - C h l o r o - 4 - [ ( 3 S ) - 3 - m e t h y l m o r p h o l i n - 4 - y l ] - 6 - ( 1 -
methylsulfonylcyclopropyl)pyrimidine (Intermediate P). Yield 78%.
1H NMR (DMSO-d6) δ 1.22 (d, 3H), 1.51 (m, 2H), 1.64 (m, 2H),
3.18 (s, 3H), 3.22 (m, 1H), 3.43 (m, 1H), 3.58 (m, 1H), 3.72 (d,
1H), 3.93 (m, 1H), 4.05 (d, 1H), 4.41 (s, 1H), 6.93 (s, 1H). ESI
(m/z): [M + H]+ = 332.
1
yield 100%). H NMR (CDCl3) δ 1.33 (3H, d), 1.42−1.51 (4H, m),
2-Chloro-4-(1-cyclopropylsulfonylcyclopropyl)-6-[(3S)-3-methyl-
1
morpholin-4-yl]pyrimidine (Intermediate AD). Yield 93%. H NMR
2.07−2.14 (2H, m), 2.40 (1H, s), 3.28−3.32 (2H, m), 3.37−3.42
(3H, m), 3.51−3.57 (1H, m), 3.66−3.70 (1H, m), 3.77−3.80 (2H,
m), 3.99−4.02 (1H, m), 4.28−4.38 (1H, m), 6.84 (1H, s). ESI (m/
z): [M + H]+ = 376.
(DMSO-d6) δ 0.87−0.92 (2H, m), 0.99−1.04 (2H, m), 1.20 (3H, d),
1.50−1.54 (2H, m), 1.59−1.63 (2H, m), 2.89−2.96 (1H, m), 3.16−
3.24 (1H, m), 3.39−3.47 (1H, m), 3.58 (1H, dd), 3.72 (1H, d), 3.93
(1H, dd), 4.06 (1H, s), 4.38 (1H, s), 6.98 (1H, s). ESI (m/z): [M +
H]+ = 358.
2 - C h l o r o - 4 - [ ( 3 S ) - 3 - m e t h y l m o r p h o l i n - 4 - y l ] - 6 - ( 1 -
methylsulfonylcyclobutyl)pyrimidine (Intermediate R). Sodium
hydroxide (50%w/w solution) (36.0 g, 450 mmol) was added to
intermediate D (5 g, 16.35 mmol), 1,3-dibromopropane (4.98 mL,
49.05 mmol), and tetrabutylammonium bromide (0.527 g, 1.64
mmol) in toluene (200 mL). The resulting suspension was stirred at
2-Chloro-4-[1-(4-fluorophenyl)sulfonylcyclopropyl]-6-[(3S)-3-
1
methylmorpholin-4-yl]pyrimidine (Intermediate AE). Yield 90%. H
NMR (DMSO-d6) δ 1.15 (3H, d), 1.53−1.56 (2H, m), 1.82−1.85
(2H, m), 3.14 (1H, dt), 3.40 (1H, dt), 3.55 (1H, dd), 3.70 (1H, d),
T
DOI: 10.1021/jm501778s
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Journal of Medicinal Chemistry
Article
45 °C for 3 h and then allowed to cool. The reaction was washed
with water, dried over MgSO4, filtered, and evaporated. The crude
product was purified by flash silica chromatography, elution gradient
30−50% ethyl acetate in DCM, to give the desired material as a solid
4.45 (3H, s), 5.53 (2H, d), 6.58−6.61 (2H, m), 6.66 (1H, s), 8.03−
8.07 (2H, m). ESI (m/z): [M + H]+ = 389.
4-[4-[(3S)-3-Methylmorpholin-4-yl]-6-(2-methylsulfonylpropan-
2-yl)pyrimidin-2-yl]aniline (Intermediate O). Yield 68%. 1H NMR
(DMSO-d6) δ 1.33 (3H, d), 1.87 (6H, s), 2.93 (3H, s), 3.28−3.35
(1H, m), 3.56−3.63 (1H, m), 3.72−3.76 (1H, m), 3.82 (1H, d), 3.90
(2H, s), 4.01−4.05 (1H, m), 4.11−4.15 (1H, m), 4.46−4.53 (1H,
m), 6.55 (1H, s), 6.71 (2H, d), 8.21 (2H, d). ESI (m/z): [M + H]+
= 391.
1
(1.89 g, yield 33%). H NMR (DMSO-d6) δ 1.22 (3H, s), 1.86−1.93
(1H, m), 2.01−2.09 (1H, m), 2.66−2.75 (2H, m), 2.84 (2H t, J=
14.7 Hz), 2.86 (3H, s), 3.18−3.25 (1H, m), 3.42−3.49 (1H, m),
3.58−3.62 (1H, m), 3.73 (1H d, J= 11.6 Hz), 3.92−3.96 (1H, m),
4.08−4.10 (1H, m), 4.43 (1H, s), 6.83 (1H, s). ESI (m/z): [M +
H]+ = 346.
4 - [ 4 - [ ( 3 S ) - 3 - M e t h y l m o r p h o l i n - 4 - y l ] - 6 - ( 1 -
methylsulfonylcyclopropyl)pyrimidin-2-yl]aniline (Intermediate Q).
2 - C h l o r o - 4 - [ ( 3 S ) - 3 - m e t h y l m o r p h o l i n - 4 - y l ] - 6 - ( 1 -
methylsulfonylcyclopentyl)pyrimidine (Intermediate T). Tetrabuty-
lammonium bromide (0.637 g, 1.98 mmol) was added to
intermediate D (6.04 g, 19.75 mmol), 1,4-dibromobutane (11.79
mL, 98.76 mmol), and sodium hydroxide solution (50% w/w) (35
mL, 438 mmol) in toluene (150 mL) under nitrogen. The resulting
mixture was stirred at 60 °C for 1.5 h before being allowed to cool
and partitioned between ethyl acetate and water. The organics were
dried over Na2SO4, concentrated in vacuo, and purified by flash silica
chromatography, eluting with 5−50% ethyl acetate in isohexane, to
1
Yield 91%. H NMR (DMSO-d6) δ 1.24 (3H, d), 1.55 (2H, m), 1.67
(2H, m), 3.23 (1H, m), 3.27 (3H, s), 3.47 (1H, m), 3.63 (1H, m),
3.77 (1H, d), 3.97 (1H, m), 4.24 (1H, s), 4.58 (1H, s), 5.75 (1H, s),
6.68 (2H, d), 8.04 (2H, d). ESI (m/z): [M + H]+ = 389.
4 - [ 4 - [ ( 3 S ) - 3 - M e t h y l m o r p h o l i n - 4 - y l ] - 6 - ( 1 -
methylsulfonylcyclobutyl)pyrimidin-2-yl]aniline (Intermediate S).
Yield 84%. 1H NMR (DMSO-d6) δ 1.21 (3H,d), 1.85−1.95
(2H,m), 2.0−2.10 (2H,m), 2.71−2.82 (2H,m), 2.82 (3H,s), 3.18
(1H,dd), 3.50 (1H,dd), 3.62 (1H,dd), 3.75 (1H,d), 3.95 (1H,dd),
4.20 (1H,d), 4.53 (1H,s), 5.55 (2H,s), 6.60 (3H,d), 8.05 (2H,d). ESI
(m/z): [M + H]+ = 403.
1
give the desired material as a white solid (5.70 g, yield 80%). H
NMR (DMSO-d6) δ 1.20 (3H, d), 1.50−1.60 (2H, m), 1.72−1.82
(2H, m), 2.30−2.41 (2H, m,), 2.50−2.60 (2H, m), 2.88 (3H, s), 3.20
(1H, dd), 3.45 (1H, dd), 3.60 (1H, dd), 3.71 (1H, d), 3.94 (1H,
dd), 4.0−4.10 (1H, m), 4.42 (1H, s), 6.89 (1H, s). ESI (m/z): [M +
H]+ = 360.
4 - [ 4 - [ ( 3 S ) - 3 - M e t h y l m o r p h o l i n - 4 - y l ] - 6 - ( 1 -
methylsulfonylcyclopentyl)pyrimidin-2-yl]aniline (Intermediate U).
1
Yield 92%. H NMR (DMSO-d6) δ 1.21 (3H,d), 1.50−1.60 (2H,m),
1.75−1.90 (2H,m), 2.34−2.43 (2H,m), 2.62−2.78 (2H,m), 2.88
(3H,s), 3.18 (1H,dd), 3.48 (1H,dd), 3.65 (1H,dd), 3.75 (1H,dd),
3.95 (1H,dd), 4.20 (1H,d), 4.51 (1H,s), 5.55 (2H,s), 6.62 (2H,d),
6.68 (1H,s), 8.09 (1H,d). ESI (m/z): [M + H]+ = 417.
2-Chloro-4-[(3S)-3-methylmorpholin-4-yl]-6-(4-methylsulfonylox-
an-4-yl)pyrimidine (Intermediate V). Sodium tert-butoxide (1.38 g,
14.39 mmol) was added portionwise to a mixture of intermediate D
(2.00 g, 6.54 mmol) and bis(2-bromoethyl) ether (2.055 mL, 16.35
mmol) in DMF (75 mL) at 0 °C over a period of 10 min under
nitrogen. The resulting solution was allowed to warm to rt and
stirred for 7 h. Further sodium tert-butoxide (629 mg, 6.54 mmol)
was added portionwise, and the solution was stirred at rt for a
further 45 h. The reaction mixture was concentrated, diluted with
ethyl acetate (200 mL), and washed sequentially with water (2 × 200
mL) and saturated brine (100 mL). The organic layer was dried
(MgSO4), filtered, and evaporated. The crude product was purified
by flash silica chromatography, elution gradient 40−100% ethyl
acetate in isohexane. Pure fractions were evaporated to dryness and
the residue was crystallized from ethyl acetate/isohexane to afford
the desired material as a white crystalline solid (1.42 g, yield 58%).
1H NMR (CDCl3) δ 1.34 (3H, d), 2.50 (2H, m), 2.55 (2H, m), 2.73
4-[4-[(3S)-3-Methylmorpholin-4-yl]-6-(4-methylsulfonyloxan-4-
yl)pyrimidin-2-yl]aniline (Intermediate W). Yield 50%. 1H NMR
(CDCl3) δ 1.34 (3H, d), 2.53 (2H, ddd), 2.70 (3H, s), 2.72 (2H, br
d), 3.33 (1H, ddd), 3.41 (2H, ddd), 3.61 (1H, ddd), 3.76 (1H, dd),
3.83 (1H, d), 3.93 (2H, s), 4.03 (3H, m), 4.18 (1H, d), 4.45 (1H,
br.d), 6.58 (1H, s), 6.71 (2H, d), 8.18 (2H, d). ESI (m/z): [M +
H]+ = 433.5.
3-[1-[2-(4-Aminophenyl)-6-[(3S)-3-methylmorpholin-4-yl]-
pyrimidin-4-yl]cyclopropyl]sulfonylpropan-1-ol (Intermediate Z).
1
Yield 71%. H NMR (DMSO-d6) δ 1.22 (3H, d), 1.52−1.54 (2H,
m), 1.60−1.62 (2H, m), 1.90−1.97 (2H, m), 3.14−3.21 (1H, m),
3.44−3.52 (5H, m), 3.60−3.64 (1H, m), 3.75 (1H, d), 3.94−3.98
(1H, m), 4.16−4.19 (1H, m), 4.48−4.55 (1H, m), 4.67 (1H, t), 5.56
(2H, s), 6.60 (2H, d), 6.67 (1H, s), 8.04 (2H, d). ESI (m/z): [M +
H]+ = 433.
(3H, s), 3.33 (3H, m), 3.56 (1H, ddd), 3.71 (1H, dd), 3.80 (1H, d),
4.01 (4H, m), 4.31 (1H, br s), 6.62 (1H, s). ESI (m/z): [M + H]+ =
376, 378.
Intermediates E, M, O, Q, S, U, W, Z, AH, AI, AJ, and AK were
prepared from intermediates D, L, N, P, R, T, V, Y, AD, AE, AF,
and AG according to the general procedure G.
4-[4-(1-Cyclopropylsulfonylcyclopropyl)-6-[(3S)-3-methylmor-
1
pholin-4-yl]pyrimidin-2-yl]aniline (Intermediate AH). Yield 78%. H
NMR (DMSO-d6) δ 0.90−0.98 (2H, m), 0.98−1.05 (2H, m), 1.22
(3H, d), 1.52−1.59 (2H, m), 1.62−1.64 (2H, m), 2.95−3.02 (1H,
m), 3.14−3.22 (1H, m), 3.45−3.51 (1H, m), 3.61−3.65 (1H, m),
3.76 (1H, d), 3.95−3.98 (1H, m), 4.14−4.17 (1H, m), 4.48−4.51
(1H, m), 5.53 (2H, d), 6.60 (2H, d), 6.75 (1H, s), 8.03−8.06 (2H,
m). ESI (m/z): [M + H]+ = 415.
4-[4-[1-(4-Fluorophenyl)sulfonylcyclopropyl]-6-[(3S)-3-methyl-
morpholin-4-yl]pyrimidin-2-yl]aniline (Intermediate AI). Yield 78%.
1H NMR (DMSO-d6) δ 1.17 (3H, d), 1.57−1.59 (2H, m), 1.86−
1.88 (2H, m), 3.12 (1H, dt), 3.45 (1H, dt), 3.60 (1H, dd), 3.73 (1H,
d), 3.95 (1H, dd), 4.10 (1H, d), 4.38 (1H, s), 5.52 (2H, s), 6.49
(2H, d), 6.55 (1H, s), 7.41 (2H, t), 7.64 (2H, d), 7.82−7.85 (2H,
m). ESI (m/z): [M + H]+ = 469.
4-[4-[(3S)-3-Methylmorpholin-4-yl]-6-(1-pyridin-4-
ylsulfonylcyclopropyl)pyrimidin-2-yl]aniline (Intermediate AJ).
Yield 73%. 1H NMR (DMSO-d6) δ 1.17 (3H, d), 1.66 (2H, q),
1.95 (2H, q), 3.13 (1H, td), 3.18 (1H, d), 3.45 (1H, td), 3.60 (1H,
dd), 3.73 (1H, d), 3.94 (1H, dd), 4.15 (1H, d), 4.43 (1H, s), 5.51
(1H, d), 6.47 (2H, d), 6.56 (1H, s), 7.50 (2H, d), 7.76 (2H, dd),
8.85 (2H, dd). ESI (m/z): [M + H]+ = 452.
General Procedure G. A mixture of the appropriate intermediate
(1 equiv), (4-aminophenyl)boronic acid pinacol ester (1−1.5 equiv),
sodium carbonate (1−5 equiv), and dichlorobis(triphenylposphine)-
palladium (5−10 mol %) in a solvent mixture of 18% DMF, 82%
DME/water/ethanol (7:3:2) was heated at 80−95 °C under a
nitrogen atmosphere for 5 h or until the reaction was complete by
TLC or LCMS analysis. The reaction mixture was partitioned
between ethyl acetate and water, and the organic layer was washed
with water or brine, dried over MgSO4, and concentrated in vacuo.
The crude material was purified by flash silica chromatography and
where necessary by ion exchange chromatography (SCX column).
4-[4-[(3S)-3-Methylmorpholin-4-yl]-6-(methylsulfonylmethyl)-
pyrimidin-2-yl]aniline (Intermediate E). Yield 90%. 1H NMR
(DMSO-d6) δ 1.23 (3H, d), 3.31 (3H, s), 3.5 (1H, m), 3.64 (1H,
m), 3.78 (1H, m), 4.13 (1H, m), 4.49 (2H, m), 5.57 (2H, s), 6.61
(2H, d), 6.68 (1H, s), 8.08 (1H, d). ESI (m/z): [M + H]+ = 363.
4-[4-(Cyclopropylsulfonylmethyl)-6-[(3S)-3-methylmorpholin-4-
yl]pyrimidin-2-yl]aniline (Intermediate M). Yield 92%. 1H NMR
(DMSO-d6) δ 0.97−1.02 (2H, m), 1.03−1.10 (2H, m), 1.23 (3H, d),
2.81−2.87 (1H, m), 3.15−3.22 (1H, m), 3.46−3.52 (1H, m), 3.62−
3.66 (1H, m), 3.77 (1H, d), 3.96−3.99 (1H, m), 4.12−4.15 (1H, m),
4-[4-[1-(Benzenesulfonyl)cyclopropyl]-6-[(3S)-3-methylmorpho-
lin-4-yl]pyrimidin-2-yl]aniline (Intermediate AK). Yield 97%. 1H
NMR (DMSO-d6) δ 1.15−1.16 (3H, d), 1.58−1.66 (2H, m), 1.85−
1.91 (2H, m), 3.07−3.14 (1H, td), 3.41−3.48 (1H, td), 3.58−3.61
(1H, dd), 3.72−3.75 (1H, d), 3.93−3.96 (1H, dd), 4.05−4.08 (1H,
U
DOI: 10.1021/jm501778s
J. Med. Chem. XXXX, XXX, XXX−XXX

Journal of Medicinal Chemistry
Article
d), 4.33 (1H, bs), 5.50 (2H, s), 6.49−6.53 (3H, t), 7.57−7.61 (2H,
t), 7.68−7.71 (3H, m), 7.78−7.81 (2H, d). ESI (m/z): [M + H]+ =
451.
Intermediates G, I, and K were prepared from intermediates D, D,
and B, respectively, in a method analogous to general procedure G
but using (4-amino-3-methylphenyl)boronic acid, (N-methyl-4-
aminophenyl)boronic acid pinacol ester, and 4-(3-
cyclopropylureido)phenylboronic acid, respectively.
2-Methoxy-4-[4-[(3S)-3-methylmorpholin-4-yl]-6-
(methylsulfonylmethyl)pyrimidin-2-yl]aniline (Intermediate G).
Yield 71%. 1H NMR (DMSO-d6) δ1.24 (d, 3H), 3.17−3.21 (m,
1H), 3.23 (s, 3H), 3.47−3.54 (m, 1H), 3.64−3.67 (m, 1H), 3.78 (d,
1H), 3.83 (s, 3H), 3.97−4.01 (m, 1H), 4.16 (d, 1H), 4.46 (s, 3H),
5.23 (s, 2H), 6.68−6.70 (m, 1H), 6.69 (s, 1H), 7.77 (m, 1H), 7.97
(s, 1H). ESI (m/z): [M + H]+ = 393.
were dried over magnesium sulfate, filtered, and concentrated in
vacuo. The crude material was chromatographed on silica, eluting
with 5% methanol in DCM, to give the desired material as a brown
1
solid (1.25 g, yield 32%). H NMR (DMSO-d6) δ 1.23 (3H, d), 2.66
(3H, d), 3.18−3.23 (1H, m), 3.46−3.52 (1H, m), 3.62−3.66 (1H,
m), 3.76 (1H, d), 3.96−3.99 (1H, m), 4.16 (1H, d), 4.45 (2H, d),
4.49 (1H, d), 5.38 (1H, t), 6.05 (1H, q), 6.66 (1H, s), 7.46−7.48
(2H, m), 8.18−8.21 (2H, m), 8.69 (1H, s). ESI (m/z): [M + H]+ =
358.
AUTHOR INFORMATION
■
Corresponding Author
*Phone: +44 7469 032383. E-mail: kurt.pike@astrazeneca.
com.
N - M e t h y l - 4 - [ 4 - [ ( 3 S ) - 3 - m e t h y l m o r p h o l i n - 4 - y l ] - 6 -
Notes
(methylsulfonylmethyl)pyrimidin-2-yl]aniline (Intermediate I). Yield
1
The authors declare no competing financial interest.
90%. H NMR (DMSO-d6) δ1.23 (3H, d), 2.74 (3H, d), 3.06−3.17
(1H, m), 3.21 (3H, s), 3.46−3.52 (1H, m), 3.62−3.66 (1H, m), 3.77
(1H, d), 3.96−4.00 (1H, m), 4.14 (1H, d), 4.44 (2H, s), 4.46 (1H,
s), 6.14 (1H, q), 6.57−6.61 (2H, m), 6.67 (1H, s), 8.10−8.13 (2H,
m). ESI (m/z): [M + H]+ = 377.
ACKNOWLEDGMENTS
■
The authors thank Dr. Paul Johnson, John Bailey, Jennifer
Peed, Matthew Box, Greg Carr, George Hill, Gillian Lamont,
Steve Glossop, and Ciorsdaidh Watts for their help synthesiz-
ing compounds that underpinned the Free−Wilson models
described, Tom Harding and Rebecca Ellston for the
generation of the potency data used in the analyses described,
Dr. Christine Cresta for helpful discussions regarding mTOR
biology, and Anne Ertan for her help generating the small
molecule crystal structure of 15 (CCDC 1019570).
3-Cyclopropyl-1-[4-[4-(hydroxymethyl)-6-[(3S)-3-methylmorpho-
1
lin-4-yl]pyrimidin-2-yl]phenyl]urea (Intermediate K). Yield 83%. H
NMR (DMSO-d6) δ 0.40−0.44 (2H, m), 0.62−0.67 (2H, m), 1.23
(3H, d), 2.54−2.58 (1H, m), 3.17−3.22 (1H, m), 3.46−3.52 (1H,
m), 3.62−3.66 (1H, m), 3.77 (1H, d), 3.96−3.99 (1H, m), 4.15−
4.18 (1H, m), 4.45 (2H, d), 4.49 (1H, d), 5.38 (1H, t), 6.41 (1H,
d), 6.66 (1H, s), 7.46−7.50 (2H, m), 8.18−8.22 (2H, m), 8.49 (1H,
s). ESI (m/z): [M + H]+ = 384.
2 - M e t h y l - 4 - [ 4 - [ ( 3 S ) - 3 - m e t h y l m o r p h o l i n - 4 - y l ] - 6 -
(methylsulfonylmethyl)pyrimidin-2-yl]aniline (Intermediate H). 4-
Bromo-2-methylaniline (1.00 g, 5.37 mmol), potassium acetate (1.59
g, 16.1 mmol), and bis(pinacolato)diboron (1.64 g, 6.45 mmol) were
dissolved in 1,4-dioxane (20 mL), and the solution was degassed for
5 min. Dichlorobis(triphenylphosphine)palladium (264 mg, 0.32
mmol) was added and the reaction stirred at 90 °C for 4 h.
Intermediate D (1.65 g, 5.37 mmol), ethanol (1.5 mL), 2 M sodium
carbonate aqueous solution (3 mL), and dichlorobis(triphenyl-
phosphine)palladium (264 mg) were added. The reaction was
maintained at 90 °C for 18 h and then allowed to cool and the
mixture partitioned between ethyl acetate and water with insoluble
material removed by filtration. The phases were separated, and the
aqueous layer was extracted with a second portion of ethyl acetate
(15 mL). The combined organics were dried (MgSO4) and
concentrated in vacuo. The residue was chromatographed on silica,
eluting with 0−3% methanol in DCM, to give the desired compound
ABBREVIATIONS USED
■
mTOR, mammalian target of rapamycin; mTORC1, mTOR
complex 1; mTORC2, mTOR complex 2; PIKK, phosphati-
dylinositol 3-kinase related kinase; ATP, adenosine triphos-
phate; DMF, N,N-dimethylformamide; DMA, N,N-dimethyla-
cetamide; DME, 1,2-dimethoxyethane; DCM, dichlorome-
thane; DBU, 1,8-diazabicyclo[5.4.0]undec-7-ene; mCPBA, 3-
chloroperbenzoic acid; DMSO, dimethyl sulfoxide; LCMS,
liquid chromatography−mass spectrometry; HPLC, high
performance liquid chromatography; TLC, thin layer
chromatography; SCX, strong cation exchange
REFERENCES
■
1
(1) Huang, S.; Houghton, P. J. Targeting mTOR signaling for
cancer therapy. Curr. Opin. Pharmacol. 2003, 3, 371−377.
(2) Yap, T. A.; Garrett, M. D.; Walton, M. I.; Raynaud, F.; de
Bono, J. S.; Workman, P. Targeting the PI3K-AKT-mTOR
pathway:progress, pitfalls, and promises. Curr. Opin. Pharmacol.
2008, 8, 393−412.
(3) Brown, E. J.; Albers, M. W.; Shin, T. B.; Ichikawa, K.; Keith, C.
T.; Lane, W. S.; Schreiber, S. L. A mammalian protein targeted by
G1-arresting rapamycin-receptor complex. Nature 1994, 369, 756−
758.
(4) Sabatini, D. M.; Erdjument-Bromage, H.; Lui, M.; Tempst, P.;
Snyder, S. H. RAFT1: a mammalian protein that binds to FKBP12 in
a rapamycin-dependent fashion and is homologous to yeast TORs.
Cell 1994, 78, 35−43.
(5) Abraham, R. T. Phosphatidylinositol 3-kinase related kinases.
Curr. Opin. Immunol. 1996, 8, 412−418.
(6) Fingar, D. C.; Salama, S.; Tsou, C.; Harlow, E.; Blenis, J.
Mammalian cell size is controlled by mTOR and its downstream
targets S6K1 and 4EBP1/eIF4E. Genes Dev. 2002, 16, 1472−1487.
(7) Fingar, D. C.; Richardson, C. J.; Tee, A. R.; Cheatham, L.;
Tsou, C.; Blenis, J. mTOR controls cell cycle progression through its
cell growth effectors S6K1 and 4E-BP1/eukaryotic translation
initiation factor 4E. Mol. Cell. Biol. 2004, 24, 200−216.
as a beige foam (290 mg, yield 15%). H NMR (DMSO-d6) δ1.22−
1.24 (m, 3H), 3.16−3.19 (m, 1H), 3.20 (s, 3H), 3.46−3.52 (m, 1H),
3.62−3.66 (m, 1H), 3.77 (d, 1H), 3.90 (s, 1H), 3.96−4.00 (m, 1H),
4.14−4.18 (m, 1H), 4.43 (s, 2H), 4.45 (s, 1H), 5.32 (s, 2H), 6.63 (s,
1H), 6.66 (s, 1H), 7.91−7.94 (m, 2H). ESI (m/z): [M + H]+ = 377.
1-[4-[4-(Hydroxymethyl)-6-[(3S)-3-methylmorpholin-4-yl]-
pyrimidin-2-yl]phenyl]-3-methylurea (Intermediate J). 1-(4-Bromo-
phenyl)-3-methylurea (2.50 g), potassium acetate (3.21 g), and
bis(pinacolato)diboron (3.33 g) were dissolved in 1,4 dioxane (120
mL). The solution was degassed for 5 min. Then 1,1′-bis(diphenyl-
phosphino)ferrocenedichloropalladium(II) dichloromethane adduct
was added (535 mg) and the reaction was heated to 90 oC for 3
hours. Further 1,1′-bis(diphenylphosphino)ferrocene-
dichloropalladium(II) dichloromethane adduct (250 mg) was
added, and heating continued for 1 hour. Further 1,1′-bis(diphenyl-
phosphino)ferrocenedichloropalladium(II) dichloromethane adduct
(250 mg) was added, and heating continued for a further 1 hour.
Intermediate B (2.66 g), ethanol (9.5 mL), 2 M sodium carbonate
solution (27.3 mL), and 1,1′-bis(diphenylphosphino)ferrocene-
dichloropalladium(II) dichloromethane adduct (535 mg) were
added, and the heating was continued for 16 h. The reaction was
cooled and evaporated to dryness and then the residue partitioned
between ethyl acetate (250 mL) and water (100 mL). The organics
V
DOI: 10.1021/jm501778s
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Journal of Medicinal Chemistry
Article
(8) Brunn, G. J.; Williams, J.; Sabers, C.; Wiederrecht, G.;
Lawrence, J. C., Jr.; Abraham, R. T. Direct inhibition of the
signaling functions of the mammalian target of rapamycin by the
phosphoinositide 3-kinase inhibitors, wortmannin and LY294002.
EMBO J. 1996, 15, 5256−5267.
(9) Sarbassov, D. D.; Ali, S. M.; Sabatini, D. M. Growing roles for
the mTOR pathway. Curr. Opin. Cell Biol. 2005, 17, 596−603.
(10) Edinger, A. L.; Linardic, C. M.; Chiang, G. G.; Thompson, C.
B.; Abraham, R. T. Differential effects of rapamycin on mammalian
target of rapamycin signaling functions in mammalian cells. Cancer
Res. 2003, 63, 8451−8460.
(11) Sun, S. Y.; Rosenberg, L. M.; Wang, X.; Fu, H.; Khuri, F.
R.Activation of Akt and eIF4E survival pathways by rapamycin-
mediated mammalian target of rapamycin inhibition. Cancer Res.
2005, 65, 7052−7058.
(12) Cloughesy, T. F.; Yoshimoto, K.; Nghiemphu, P.; Brown, K.;
Dang, J.; Zhu, S.; Hsueh, T.; Chen, Y.; Wang, W.; Youngkin, D.;
Liau, L.; Martin, N.; Becker, D.; Bergsneider, M.; Lai, A.; Green, R.;
Oglesby, T.; Koleto, M.; Trent, J.; Horvath, S.; Mischel, P. S.;
Mellinghoff, I. K.; Sawyers, C. L. Antitumor activity of rapamycin in
(21) (a) Assessments of aqueous solubility were made after an
incubation of 24 h in pH 7.4 phosphate buffer. After centrifugation,
analysis of the supernatant liquid was performed by LC−UV to
quantify the amount of compound in solution. Further details are
contained in ref 21b. (b) Buttar, D.; Colclough, N.; Gerhardt, S.;
MacFaul, P. A.; Phillips, S. D.; Plowright, A.; Whittamore, P.; Tam,
K.; Maskos, K.; Steinbacher, S.; Steuber, H.
A combined
spectroscopic and crystallographic approach to probing drug−
human serum albumin interactions. Bioorg. Med. Chem. 2010, 18,
7486−7496.
(22) Finlay, M. R. V; Morris, J.; Pike, K. G. Morpholinopyrimdine
derivatives, processes for preparing them, pharmaceutical composi-
tions containing them, and their use for treating proliferative
disorders. PCT Int. Appl. WO2008023159, 2008.
(23) Pike, K. G. Morpholinopyrimidine derivatives that are useful in
the treatment of diseases mediated by mTOR and/or PI3K enzyme
and their preparation. PCT Int. Appl. WO2009007750, 2009.
(24) Blades, K.; Demeritt, J.; Fillery, S.; Foote, K. M.; Greenwood,
R.; Gregson, C.; Hassall, L. A.; McGuire, T. M.; Pike, K. G.;
Williams, E. Expedient synthesis of biologically important sulfonyl-
methyl pyrimidines. Tetrahedron Lett. 2014, 55 (29), 3851−3855.
(25) Pike, K. G. Pyrimidine derivatives that are useful in the
treatment of diseases mediated by mTOR and/or PI3K enzyme and
their preparation. PCT Int. Appl. WO2009007748, 2009.
(26) Pike, K. G.; Malagu, K.; Hummersone, M. G.; Menear, K. A.;
Duggan, H. M. E.; Gomez, S.; Martin, N. M. B.; Ruston, L.; Pass, S.
L.; Pass, M. Optimization of potent and selective dual mTORC1 and
mTORC2 inhibitors: The discovery of AZD8055 and AZD2014.
Bioorg. Med. Chem. Lett. 2013, 23, 1212−1216.
a
phase I trial for patient with recurrent PTEN-deficient
glioblastoma. PLoS Med. 2008, 5 (1), e8.
(13) (a) Garcia-Echeverria, C. Allosteric and ATP-competitive
kinase inhibitors of mTOR for cancer treatment. Bioorg. Med. Chem.
Lett. 2010, 20, 4308−4312. (b) Faivre, S.; Kroemer, G.; Raymond, E.
Current development of mTOR inhibitors as anticancer agents. Nat.
Rev. Drug Discovery 2006, 5, 671−688. (c) Finlay, M. R. V.; Griffin,
R. J. Modulation of DNA repair by pharmacological inhibitors of the
PIKK protein kinase family. Bioorg. Med. Chem. Lett. 2012, 22,
5352−5359. (d) Cano, C.; Saravanan, K.; Bailey, C.; Bardos, J.;
Curtin, N. J.; Frigerio, M.; Golding, B. T.; Hardcastle, I. R.;
Hummersone, M. G.; Menear, K. A.; Newell, D. R.; Richardson, C.
J.; Shea, K.; Smith, G. C. M.; Thommes, P.; Ting, A.; Griffin, R. J. 1-
Substituted (dibenzo[b,d]thiophen-4-yl)-2-morpholino-4H-chromen-
4-ones endowed with dual DNA-PK/PI3-K inhibitory activity. J.
Med. Chem. 2013, 56, 6386−6401.
(14) Walker, E. H.; Pacold, M. E.; Perisic, O.; Stephens, L.;
Hawkins, P. T.; Wymann, M. P.; Williams, R. L. Structural
determinants of phosphoinositide 3-kinase inhibition by wortmannin,
LY294002, quercetin, myricetin, and staurosporine. Mol. Cell 2000, 6,
909−919.
(15) Yang, H.; Rudge, D. G.; Koos, J. D.; Vaidialingam, B.; Yang,
H. J.; Pavletich, N. P. mTOR kinase structure, mechanism and
regulation. Nature 2013, 497, 217−223.
(16) Finlay, M. R. V.; Buttar, D.; Critchlow, S. E.; Dishington, A.
P.; Fillery, S. M.; Fisher, E.; Glossop, S. C.; Graham, M. A.; Johnson,
T.; Lamont, G. M.; Mutton, S.; Perkins, P.; Pike, K. G.; Slater, A. M.
Sulfonyl-morpholino-pyrimidines: SAR and development of a novel
class of selective mTOR kinase inhibitor. Bioorg. Med. Chem. Lett.
2012, 22, 4163−4168.
(17) Inhibition of mTOR kinase was evaluated in a high-throughput
assay using α screen capture complex technology (Perkin-Elmer)
with a recombinant truncated FLAG-tagged mTOR (amino acids
1362−2549, expressed in HEK293 cells). The activity of the lipid
kinases, class I PI3Ks α, β, δ, γ was measured with the lipid PIP2 as
substrate. More detailed description of the assay conditions are
described in ref 27.
(18) Walker, E. H.; Pacold, M. E.; Perisic, O.; Ried, C.; Stephens,
L.; Williams, R. L. Structural insights into phosphoinositide 3-kinase
catalysis and signalling. Nature 1999, 402, 313−320.
(19) Figure 2b was generated using PyMOL: DeLano, W. L. The
PyMOL Molecular Graphics System; DeLano Scientific: San Carlos,
CA, U.S., 2002; http://www.pymol.org.
(27) Chresta, C. M.; Davies, B. R.; Hickson, I.; Harding, T.;
Cosulich, S.; Critchlow, S. E.; Vincent, J. P.; Ellston, R.; Jones, D.;
Sini, P.; James, D.; Howard, Z.; Dudley, P.; Hughes, G.; Smith, L.;
Maguire, S.; Hummersone, M.; Malagu, K.; Menear, K.; Jenkins, R.;
Jacobsen, M.; Smith, G. C. M.; Guichard, S.; Pass, M. AZD8055 is a
potent, selective, and orally bioavailable ATP-competitive mammalian
target of rapamycin kinase inhibitor with in vitro and in vivo
antitumor activity. Cancer Res. 2010, 70, 288−298.
(28) (a) Hopkins, A. L.; Groom, C. R.; Alex, A. Ligand efficiency: a
useful metric for lead selection. Drug. Discovery Today 2004, 9, 430−
431. (b) Albert, J. S.; Blomger, N.; Breeze, A. L.; Brown, A. J. H.;
Burrows, J. N.; Edwards, P. D.; Folmer, R. H. A.; Geschwinder, S.;
Griffen, E. J.; Kenny, P. W.; Nowak, T.; Olssom, L.-L.; Sanganee, H.;
Shapiro, A. B. An integrated approach to fragment-based lead
generation: philosophy, strategy and case studies from AstraZeneca’s
drug discovery programmes. Curr. Top. Med. Chem. 2007, 7, 1600−
1629. (c) Edwards, M. P.; Price, D. A. Role of physicochemical
properties and ligand lipophilicity efficiency in addressing drug safety
risks. Annu. Rep. Med. Chem. 2010, 45, 381−391. (d) Tarcsay, A.;
Nyiri, K.; Keseru, G. M. Impact of lipophilic efficency on compound
quality. J. Med. Chem. 2012, 55, 1252−1260. (e) Chiou, W. L.;
Robbiel, G.; Chung, S. M.; Wul, T.-C.; Mal, C. Pharm. Res. 1998, 15,
1474. (f) Pike, K. G.; Allen, J. V.; Caulkett, P. W. R.; Clarke, D. S.;
Donald, C. S.; Fenwick, M. L.; Johnson, K. M.; Johnstone, C.;
McKerrecher, D.; Rayner, J. W.; Walker, R. P.; Wilson, I. Design of a
potent, soluble glucokinase activator with increased pharmacokinetic
half-life. Bioorg. Med. Chem. Lett. 2011, 21, 3467−3470. (g) Murray,
C. W.; Erlanson, D. A.; Hopkins, A. L.; Keseru, G. M.; Leeson, P. D.;
Rees, D. C.; Reynolds, C. H.; Richmond, N. J. Validity of ligand
efficiency metrics. Med. Chem. Lett. 2014, 5, 616−618.
(29) Zask, A.; Kaplan, J.; Verheijen, J. C.; Richard, D. J.; Curran,
K.; Brooijmans, N.; Bennett, E. M.; Toral-Barza, L.; Hollander, I.;
Ayral-Kaloustian, S.; Yu, K. Morpholine derivatives greatly enhance
the selectivity of mammalian target of rapamycin (mTOR) inhibitors.
J. Med. Chem. 2009, 52, 7942−7945.
(30) Foote, K. M.; Blades, K.; Cronin, A.; Filery, S.; Guichard, S. S.;
Hassall, L.; Hickson, I.; Jacq, X.; Jewsbury, P. J.; McGuire, T. M.;
Nissink, J. W. M.; Odedra, R.; Page, K.; Perkins, P.; Suleman, A.;
Tam, K.; Thommes, P.; Broadhurst, R.; Wood, C. Discovery of 4-{4-
[(3R)-3-methylmorpholin-4-yl]-6-[1-(methylsulfonyl)cyclopropyl]-
(20) MDA-MB-468 cells were exposed for 2 h to increasing
concentrations of compound. At the end of the incubation period
cells were fixed, washed, and probed with antibodies against S473
pAKT or against S235/236 phosphorylated S6 (pS6). Levels of
phosphorylation were assessed using an Acumen laser scanning
cytometer (TTP Labtech).
W
DOI: 10.1021/jm501778s
J. Med. Chem. XXXX, XXX, XXX−XXX

Journal of Medicinal Chemistry
Article
pyrimidin-2-yl}-1H-indole (AZ20): a potent and selective inhibitor of
ATR protein kinase with monotherapy in vivo antitumor activity. J.
Med. Chem. 2013, 56, 2125−2138.
(31) Ganellin, R. 1980 Award in medicinal chemistry. Medicinal
chemistry and dynamic structure−activity analysis in the discovery of
drugs acting at histamine H2 receptors. J. Med. Chem. 1981, 24,
913−920.
(32) (a) Free, S. M., Jr.; Wilson, J. W. A mathematical contribution
to structure−activity studies. J. Med. Chem. 1964, 7, 395−399.
(b) Freeman-Cook, K. D.; Amor, P.; Bader, S.; Buzon, L. M.; Coffey,
S. B.; Corbett, J. W.; Dirico, K. J.; Dorn, S. D.; Elliott, R. L.; Esler,
W.; Guzman-Perez, A.; Henegar, K. E.; Houser, J. A.; Jones, C. S.;
Limberakis, C.; Loomis, K.; McPherson, K.; Murdande, S.; Nelson,
K. L.; Phillion, D.; Pierce, B. S.; Song, W.; Sugarman, E.; Tapley, S.;
Tu, M.; Zhao, Z. Maximising lipophilic efficiency: the use of Free−
Wilson analysis in the design of inhibitors of acetyl-CoA carboxylase.
J. Med. Chem. 2012, 55, 935−942. (c) Goldberg, F. W.; Leach, A. G.;
Scott, J. S.; Snelson, W. L.; Groombridge, S. D.; Donald, C. S.;
Bennett, S. N. L.; Bodin, C.; Morentin Gutierrez, P.; Gyte, A. C.
Free−Wilson and structural approaches to co-optimizing human and
rodent isoform potency for 11β-hydroxysteriod dehydrogenase type 1
(11β-HSD1) inhibitors. J. Med. Chem. 2012, 55, 10652−10661.
(33) Waring, M. J.; Johnstone, C.; McKerrecher, D.; Pike, K. G.;
Robb, G. Matrix-based multiparameter optimisation of glucokinase
activators: the discovery of AZD1092. Med. Chem. Commun. 2011, 2,
775−779.
(34) (a) Monolayers of MDCKII (Madin−Darby canine kidney
cells) cells transfected with the human MDR1 (multidrug resistance
1) transporter protein were cultured in Transwells and used to study
the permeability and efflux potential of compounds at
a
concentration of 10 μM.. (b) Keogh, J. P.; Kunta, J. R. Development,
validation and utility of an in vitro technique for assessment of
potential clinical drug−drug interactions involving P-glycoprotein.
Eur. J. Pharm. Sci. 2006, 27, 543−554.
(35) Mouse clearance values were obtained following an iv dose of
2.5−10 μmol/kg to CD-1 outbred mice, and dog clearance values
were obtained following an iv dose of 1−3 μmol/kg to Beagle dogs.
The compounds were dosed as a solution, and the clearance values
are quoted in mL min−1 kg−1. Bioavailability values were calculated
following an oral dose of 5−20 μmol/kg in mouse and 3−10 μmol/
kg in dog.
(36) Redfern, W. S.; Carlsson, L.; Davis, A. S.; Lynch, W. G.;
MacKenzie, I.; Palethorpe, S.; Siegl, P. K. S.; Strang, I.; Sullivan, A.
T.; Wallis, R.; Camm, A. J.; Hammond, T. G. Relationships between
preclinical cardiac electrophysiology, clinical QT interval prolonga-
tion and torsade de pointes for a broad range of drugs: evidence for
a provisional safety margin in drug development. Cardiovasc. Res.
2003, I, 32−45.
(37) The plasma protein binding of compounds was measured
using equilibrium dialysis technique and is quoted as the fraction
unbound (fu). A more detailed description of the equilibrium dialysis
assay conditions are described in Chiou, W. L.; Robbiel, G.; Chung,
S. M.; Wul, T.-C.; Mal, C. Correlation of plasma clearance of 54
extensively metabolized drugs between humans and rats: mean
allometric coefficient of 0.66. Pharm. Res. 1998, 15, 1474−1479.
X
DOI: 10.1021/jm501778s
J. Med. Chem. XXXX, XXX, XXX−XXX