Note: Descriptions are shown in the official language in which they were submitted.
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SPIRO BICYCLIC INHIBITORS OF MENIN-MLL INTERACTION
FIELD OF THE INVENTION
The present invention relates to pharmaceutical agents useful for therapy
and/or
prophylaxis in a mammal, and in particular to spiro bicyclic compounds,
pharmaceutical
composition comprising such compounds, and their use as menin/MLL
protein/protein
interaction inhibitors, useful for treating diseases such as cancer,
myelodysplastic
syndrome (MDS) and diabetes.
BACKGROUND OF THE INVENTION
Chromosomal rearrangements affecting the mixed lineage leukemia gene (MLL;
MLL1;
KMT2A) result in aggressive acute leukemias across all age groups and still
represent
mostly incurable diseases emphasizing the urgent need for novel therapeutic
approaches.
Acute leukemias harboring these chromosomal translocations of MLL represent as
lymphoid, myeloid or biphenotypic disease and constitute 5 to 10% of acute
leukemias
in adults and approximately 70% in infants (Marschalek, Br J Haematol 2011.
152(2),
141-54; Tomizawa et al., Pediatr Blood Cancer 2007. 49(2), 127-32).
MLL is a histone methyltransferase that methylates histone H3 on lysine 4
(H3K4) and
functions in multiprotein complexes. Use of inducible loss-of-function alleles
of Mill
demonstrated that M111 plays an essential role in sustaining hematopoietic
stem cells
(HSCs) and developing B cells although its histone methyltransferase activity
is
dispensable for hematopoiesis (Mishra et al., Cell Rep 2011. 7(4), 1239-47).
Fusion of MLL with more than 60 different partners has been reported to date
and has
been associated with leukemia formation/progression (Meyer et al., Leukemia
2013.
27, 2165-2176). Interestingly, the SET (Su(var)3-9, enhancer of zeste, and
trithorax) domain of MLL is not retained in chimeric proteins but is replaced
by the
fusion partner (Thiel et al., Bioessays 2012. 34, 771-80). Recruitment of
chromatin
modifying enzymes like Dot1L and/or the pTEFb complex by the fusion partner
leads to
enhanced transcription and transcriptional elongation of MLL target genes
including
HOXA genes (e.g. HOXA9) and the HOX cofactor MEIS1 as the most prominent ones.
Aberrant expression of these genes in turn blocks hematopoietic
differentiation and
enhances proliferation.
Menin which is encoded by the Multiple Endocrine Neoplasia type 1 (MEN]) gene
is
expressed ubiquitously and is predominantly localized in the nucleus. It has
been shown
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to interact with numerous proteins and is, therefore, involved in a variety of
cellular
processes. The best understood function of menin is its role as an oncogenic
cofactor of
MLL fusion proteins. Menin interacts with two motifs within the N-terminal
fragment of
MLL that is retained in all fusion proteins, MBM1 (menin-binding motif 1) and
MBM2
(Thiel et al., Bioessays 2012. 34, 771-80). Menin/MLL interaction leads to the
formation
of a new interaction surface for lens epithelium-derived growth factor
(LEDGF).
Although MLL directly binds to LEDGF, menin is obligatory for the stable
interaction
between MLL and LEDGF and the gene specific chromatin recruitment of the MLL
complex via the PWWP domain of LEDGF (Cermakova et al., Cancer Res 2014. 15,
5139-51; Yokoyama & Cleary, Cancer Cell 2008. 8, 36-46). Furthermore, numerous
genetic studies have shown that menin is strictly required for oncogenic
transformation
by MLL fusion proteins suggesting the menin/MLL interaction as an attractive
therapeutic target. For example, conditional deletion of Men] prevents
leukomogenesis
in bone marrow progenitor cells ectopically expressing MLL fusions (Chen et
al., Proc
Natl Acad Sci 2006. 103, 1018-23). Similarly, genetic disruption of menin/MLL
fusion
interaction by loss-of-function mutations abrogates the oncogenic properties
of the MLL
fusion proteins, blocks the development of leukemia in vivo and releases the
differentiation block of MLL-transformed leukemic blasts. These studies also
showed
that menin is required for the maintenance of HOX gene expression by MLL
fusion
proteins (Yokoyama et al., Cell 2005. 123, 207-18). In addition, small
molecule
inhibitors of menin/MLL interaction have been developed suggesting
druggability of this
protein/protein interaction and have also demonstrated efficacy in preclinical
models of
AML (Borkin et al., Cancer Cell 2015. 27, 589-602; Cierpicki and Grembecka,
Future
Med Chem 2014. 6, 447-462). Together with the observation that menin is not a
requisite
cofactor of MLL1 during normal hematopoiesis (Li et al., Blood 2013. 122, 2039-
2046),
these data validate the disruption of menin/MLL interaction as a promising new
therapeutic approach for the treatment of MLL rearranged leukemia and other
cancers
with an active HOXIMEIS1 gene signature. For example, an internal partial
tandem
duplication (PTD) within the 5 'region of the MLL gene represents another
major
aberration that is found predominantly in de novo and secondary AML as well as
myeloid
dysplasia syndromes. Although the molecular mechanism and the biological
function of
MLL-PTD is not well understood, new therapeutic targeting strategies affecting
the
menin/MLL interaction might also prove effective in the treatment of MLL-PTD-
related
leukemias. Furthermore, castration-resistant prostate cancer has been shown to
be
dependent on the menin/MLL interaction (Malik et al., Nat Med 2015. 21, 344-
52).
Several references describe inhibitors targeting the menin-MLL interaction:
W02011029054, J Med Chem 2016, 59, 892-913 describes the preparation of
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thienopyrimidine and benzo diazepine derivatives; W02014164543 describes
thienopyrimidine and thienopyridine derivatives; Nature Chemical Biology March
2012,
8, 277-284 and Ren, J.; et at. Bioorg Med Chem Lett (2016),
http ://dx.doi.org/10.1016/j .bmc1.2016.07.074 describe thienopyrimidine
derivatives; J
Med Chem 2014, 57, 1543-1556 describes hydroxy- and aminomethylpiperidine
derivatives; and Future Med Chem 2014, 6, 447-462 reviews small molecule and
peptidomimetic compounds. W02017112768 describes inhibitors of the menin-MLL
interaction.
DESCRIPTION OF THE INVENTION
The present invention concerns novel compounds of Formula (I):
, 2
L11- X 3
R
1
R \ _______ el
S'N R2
(I),
and the tautomers and the stereoisomeric forms thereof, wherein
Rl is selected from the group consisting of CH3, CH2F, CHF2, and CF3;
R2 is selected from the group consisting of hydrogen and CH3;
Ll represents a 7- to 10-membered saturated spiroheterobicyclic system
containing one
or two N-atoms provided that it is N-linked to the thienopyrimidinyl
heterocycle; and
--L2-R3 is selected from (a), (b), (c), (d), (e), (f) or (g), wherein
(a) L2 is selected from the group consisting of >S02, >CR4aR4b, and -CHR4aCHR5-
;
wherein
(0 when L2 is linked to a carbon atom of Ll, then R4a and R5 are
each
independently selected from the group consisting of hydrogen; -0R6;
-NR7aR7b; -C(=0)NR7aR7b; C1_4alkyl optionally substituted with a
substituent selected from the group consisting of fluoro, -CN, -0R8, and
-NR9aR9b; and C-linked 4- to 7-membered non-aromatic heterocyclyl
containing at least one nitrogen, oxygen or sulfur atom;
(ii) when L2 is linked to a nitrogen atom of Ll, then R4a is
selected from the
group consisting of hydrogen; -C(=0)NR7aR7b; C1_4alkyl optionally
substituted with a substituent selected from the group consisting of fluoro,
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-CN, -0R8, and -NR9aR9b; and C-linked 4- to 7-membered non-aromatic
heterocyclyl containing at least one nitrogen, oxygen or sulfur atom;
R5 is selected from the group consisting of hydrogen; -0R6; -NR7aR7b;
-C(=0)NR7aR7b; Ci_4alkyl optionally substituted with a substituent
selected from the group consisting of fluoro, -CN, -0R8, and
-NR9aR9b; and C-linked 4- to 7-membered non-aromatic heterocyclyl
containing at least one nitrogen, oxygen or sulfur atom;
R4b is selected from the group consisting of hydrogen and methyl; or R4a and
R4b
together with the carbon atom to which they are attached form a C3_5cycloalkyl
or a C-linked 4- to 6-membered heterocyclyl containing an oxygen atom; wherein
R6, R7a, R7b, R8, R9a and R9b are each independently selected from the group
consisting of hydrogen; Ci_4a1ky1 optionally substituted with a substituent
selected from the group consisting of fluoro, -CN, and -C(=0)NR10aRlOb; and C2-
4alkyl substituted with a substituent selected from the group consisting of
-0R11 and -NR10aRlOb; wherein
gioa, glob and R"
are each independently selected from the group consisting of
hydrogen; Ci_4a1ky1; and C-linked 4- to 7-membered non-aromatic heterocyclyl
containing at least one nitrogen, oxygen or sulfur atom; and
R3 is selected from the group consisting of Ar, Het', Het2, and a 7- to 10-
membered saturated spirocarbobicyclic system; or
(b) L2 is selected from the group consisting of >cg4cg4d and -CHR4cCHR5a-,
wherein
g4c5 g4a5 and R5a are each independently selected from the group consisting of
hydrogen and Ci_4a1ky1; and
R R
12a 12a
I I
.si Ri2b ,Ge _Ri 2b
-- '12c
R3 is selected from the group consisting of R and R ;
wherein R12a, gin, and R12c are each independently selected from the group
consisting of Ci_6a1ky1 optionally substituted with a ¨OH or a ¨NH2
substituent;
and -0Ci_6a1ky1; or
(c) --L2-R3 is Ci_6a1ky1 optionally substituted with one, two or three fluoro
substituents;
or
(d) L2 is 0 and R3 is selected from the group consisting of C3_6alkyl
optionally
substituted with one, two or three fluoro substituents; Ar; Het'; Het2; a 7-
to 10-
membered saturated spirocarbobicyclic system; -CH2-Ar; -CH2-Het'; -CH2-Het2;
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and -CH2-(a 7- to 10-membered saturated spirocarbobicyclic system); when L2 is
linked to a carbon atom of Ll; or
(e) --L2-R3 is ¨0-CHR5-R3 when L2 is linked to a carbon atom of Ll,
wherein
R5 is selected from the group consisting of -C(=0)NR13aRl3b;
Ci_4a1ky1 optionally substituted with a substituent selected from the group
consisting of fluoro, -OR", and -NR15aRl5b; and C-linked 4-to 7-membered non-
aromatic heterocyclyl containing at least one nitrogen, oxygen or sulfur atom;
wherein
R13a, R13b5 R145 R15a and Risb are each independently selected from the group
consisting of hydrogen; Ci_4a1ky1 optionally substituted with a substituent
selected from the group consisting of fluoro and -C(=0)NR16aRl6b; and
C2_4alkyl substituted with a substituent selected from the group consisting of
-OR' and -NR16aRl6b; wherein
R16a5 R16b and K-17
are each independently selected from the group consisting of
hydrogen; Ci_zialkyl; and C-linked 4- to 7-membered non-aromatic heterocyclyl
containing at least one nitrogen, oxygen or sulfur atom; and
R3 is selected from the group consisting of hydrogen; Ci_4a1ky1 optionally
substituted with one, two, or three fluoro substituents; -CN; Ar, Het'; Het2;
and a
7- to 10-membered saturated spirocarbobicyclic system; or
R18a
R181::. )........0
N
I 20 (f) --L2-R3 is R18 5 wherein
R" is selected from the group consisting of hydrogen; Ci_4a1ky1 optionally
substituted with a fluoro or a ¨CN substituent; and C2_4alkyl substituted with
a
substituent selected from the group consisting of ¨OR' and ¨NR2 aR
20b; wherein
V, R20a and R2 b are each independently selected from the group consisting of
hydrogen; Ci_4a1ky1 optionally substituted with a substituent selected from
the
group consisting of fluoro, -CN, and -C(=0)NR2laR21b; C2_4alkyl substituted
with
a substituent selected from the group consisting of ¨0R22 and _NR2iaR21b; and
C-
linked 4- to 7-membered non-aromatic heterocyclyl containing at least one
nitrogen, oxygen or sulfur atom; wherein
R21a5 R2lb and K-.22
are each independently selected from the group consisting of
hydrogen and Ci_zialkyl; and
R"a is selected from the group consisting of hydrogen, fluoro and Ci_zialkyl;
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Ri" is selected from the group consisting of fluoro, -0C1_4alkyl, and
C1_4alkyl
optionally substituted with 1, 2 or 3 fluoro substituents; or
R18a and Ri8b are bound to the same carbon atom and together form a
C3_5cycloalkyl or a C-linked 4- to 6-membered heterocyclyl containing an
oxygen
atom; or
N-
---.... S
-. -'
N
(g) --L2-R3 is or ; and wherein
Ar is phenyl or naphthyl, each of which may be optionally substituted with
one, two, or
three substituents each independently selected from the group consisting of
halo, -CN,
io _0R245 _NR25aR25b5 and C1_4alkyl optionally substituted with a
substituent selected from
the group consisting of fluoro, -CN, -0R26, -NR27aR27b, and ¨C(=0)NR27aR27b;
Het' is a monocyclic heteroaryl selected from the group consisting of pyridyl,
4-, 5- or
6-pyrimidinyl, pyrazinyl, pyridazinyl, furanyl, thienyl, pyrrolyl, pyrazolyl,
imidazolyl,
4- or 5-thiazolyl, isothiazolyl, thiadiazolyl, and isoxazolyl; or a bicyclic
heteroaryl
selected from the group consisting of imidazothiazolyl, imidazoimidazolyl,
benzofuranyl, benzothiophenyl, benzimidazolyl, benzoxazo lyl, isobenzoxazo
lyl,
benziso xazo lyl, benzothiazo lyl, benzisothiazo lyl, isobenzofuranyl, indo
lyl, iso indolyl,
indolizinyl, indolinyl, isoindolinyl, indazo lyl, pyrazolopyridinyl,
pyrazolopyrimidinyl,
imidazopyridinyl, imidazopyrazinyl, imidazopyridazinyl; each of which may be
optionally substituted with one, two, or three substituents each independently
selected
5 _NR25aR25b5
from the group consisting of halo, -CN, _0R24 and
C1_4alkyl optionally
substituted with a substituent selected from the group consisting of fluoro, -
CN, -0R26, -
NR27aR27b, and ¨C(=0)NR27aR27b; and
Het2 is a non-aromatic heterocyclyl optionally substituted with one, two, or
three
substituents each independently selected from the group consisting of halo, -
CN, -OR',
-NR25aR25b, and C1_4alkyl optionally substituted with a substituent selected
from the
group consisting of fluoro, -CN, -0R26, -NR27aR27b, and ¨C(=0)NR27aR27b;
wherein
R24, R25a, R25b, R26, R27a, and R27b are each independently selected from the
group
consisting of hydrogen; C1_4a1ky1 optionally substituted with a substituent
selected from
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the group consisting of fluoro and -C(=0)NR28aR28b; and C2_4a1ky1 substituted
with a
substituent selected from the group consisting of -OR' and ¨NR28aR28b; wherein
R28a, R28b and K-29
are each independently selected from the group consisting of hydrogen;
C1_4alkyl; and C-linked 4- to 7-membered non-aromatic heterocyclyl containing
at least
one nitrogen, oxygen or sulfur atom;
and the pharmaceutically acceptable salts and the solvates thereof;
provided that the following compounds, and pharmaceutically acceptable
addition salts,
and solvates thereof are excluded:
0 r" (Co _________
N N
N
...N1
N
(----..)
_--X
/ I N
F A_Cl F / F s....õ.
S N S N F
F F F
F F
Y
N
p rc
N N
N
N
/
F I ....-N--....
S N
F I 1
F / I N F /
S
F,../.. F
S-----N F
F
F
(CO /4---
rl<F
N Ns
y N
N
.........N
...:N
...:N
N
F{
F
/ F/ I /
I N N F I
S
S N S N F
F
F
F
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0 H _______
r<
H N
1/0
N N CN
SZ N
SZ
1\1
N
/ I NI / N
F/ / I LN F I
S-----N F/ s N
S---"N F
f
FiF F
FF
rL. _ CN
N H IP
X
N
N
F)c(---X l
/ I N c---/L"
F / 1
s N S-."-N
F F
F F
eiN
F
F S---"N
F
----( )------\ >r\
S-1(
N
X oN
C
N C
N
/ ---)
/cl
c
/ "
1 " / 1 c(XLN
F(*-X
S N F
F FiF
F S N
F
F
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ri * H
N
\ NC
--
N H N
N
N
X
F I
S N
C----)N
F
F F / 1 C----)N
sN
FF FV se
F7F
CN CN
rCO
_ --
N H NH
0 N
N ( )N
./I
F
6----)N S N
N F/
F
N)1-----_\
I \ N,j'X'__\,
N S CF3 I
N S CF3
* CN
ON
0 CN ____
NH
N N
(.1
( )N
(i----)N
N
e"-----", N
F/
S---N! / 1
FV se
/-'
F/F
N
N 1 \
N S C F3
H H
/
0 N
0 N
101 00 ON
0
H
( ... .) H
N
N
N
C9N
C9N
F,,/
S N:- F
--",-
Fr--F ' I -j / I
/ F
sN
c--.)
F/---F
Fr.'F
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0
,>=o
*
i NH2
9N N
(
(9
N N
(i----)N
C C"-
N )N
' 1 F s_N F, /
F
F / 1
S----N- F F ?'F S---N
F
F
0 CN rfi
H -
N H
N
N
8 N
8
N
8 N
C----)N
/
F/
F / S I
----NJ I N F s__,..N1
Fi
F F
F
S N
F
F
NC
0
N ---
\ ) \
6 b H N
N 6
N
N
/ I
/
F
FiF S--"'"N F./
F I
S F ---N- (----)1 N
scF7
FiF
H
'N
6 NC
N
\ 0
N
Si
N
/ \
N
N
(-N ---/L N
F/
F7 I
S----N
F)c('- N
I ) F
F
F
F / C-rSN
S N
F
F
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N
00
i1 nN
nN
N N
N
N F/Ces--XLI F)c(*---XL
/ I N
F(----X
F /cL
/ I
S N S N
F
F F S
F
F
<\0
6 cc
/ 1
(
1
N
1
1\1----- c, \ 1\11 \ L-N--"----0 CF3
I \
N S CF3
N S CF3
NC
= ).-----\
,--'
H N N
oN
QN
6----:1
0 NI-------. -D
L I \ CF3
HN N-----S
H
F/
/ I
S--"N F./
S----N
FiF F/F%
\
/ ( / ( 7
iN iN iN =
&N) 0
1\1---- N
\ L 1 \ N----S \OF
3 N Q µ-' CF3
N S CF3
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NC
FIN 0
(I)
N
/ N
F/CC---1
S N
F
F
It is to be understood that the above proviso applies to all embodiments of
the present
invention described hereinafter.
The present invention also relates to a pharmaceutical composition comprising
a
.. therapeutically effective amount of a compound of Formula (I), a
pharmaceutically
acceptable salt, or a solvate thereof, and a pharmaceutically acceptable
carrier or
excipient.
Additionally, the invention relates to a compound of Formula (I), a
pharmaceutically
acceptable salt, or a solvate thereof, for use as a medicament, and to a
compound of
Formula (I), a pharmaceutically acceptable salt, or a solvate thereof, for use
in the
treatment or in the prevention of cancer, myelodysplastic syndrome (MDS) and
diabetes.
In a particular embodiment, the invention relates to a compound of Formula
(I), a
pharmaceutically acceptable salt, or a solvate thereof, for use in the
treatment or in the
prevention of cancer.
In a specific embodiment said cancer is selected from leukemias, myeloma or a
solid
tumor cancer (e.g. prostate cancer, lung cancer, breast cancer, pancreatic
cancer, colon
cancer, liver cancer, melanoma and glioblastoma, etc.). In some embodiments,
the
leukemias include acute leukemias, chronic leukemias, myeloid leukemias,
myelogeneous leukemias, lymphoblastic leukemias, lymphocytic leukemias, Acute
myelogeneous leukemias (AML), Chronic myelogenous leukemias (CML), Acute
lymphoblastic leukemias (ALL), Chronic lymphocytic leukemias (CLL), T cell
prolymphocytic leukemias (T-PLL), Large granular lymphocytic leukemia, Hairy
cell
leukemia (HCL), MLL-rearranged leukemias, MLL-PTD leukemias, MLL amplified
leukemias, MLL-positive leukemias, leukemias exhibiting HOXIMEIS1 gene
expression
signatures etc.
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The invention also relates to the use of a compound of Formula (I), a
pharmaceutically
acceptable salt, or a solvate thereof, in combination with an additional
pharmaceutical
agent for use in the treatment or prevention of cancer, myelodysplastic
syndrome (MDS)
and diabetes.
Furthermore, the invention relates to a process for preparing a pharmaceutical
composition according to the invention, characterized in that a
pharmaceutically
acceptable carrier is intimately mixed with a therapeutically effective amount
of a
compound of Formula (I), a pharmaceutically acceptable salt, or a solvate
thereof
The invention also relates to a product comprising a compound of Formula (I),
a
pharmaceutically acceptable salt, or a solvate thereof, and an additional
pharmaceutical
agent, as a combined preparation for simultaneous, separate or sequential use
in the
treatment or prevention of cancer, myelodysplastic syndrome (MDS) and
diabetes.
Additionally, the invention relates to a method of treating or preventing a
cell
proliferative disease in a warm-blooded animal which comprises administering
to the
said animal an effective amount of a compound of Formula (I), a
pharmaceutically
acceptable salt, or a solvate thereof, as defined herein, or a pharmaceutical
composition
or combination as defined herein.
DETAILED DESCRIPTION OF THE INVENTION
The term 'halo' or 'halogen' as used herein represents fluoro, chloro, bromo
and iodo.
The prefix `Cx_y' (where x and y are integers) as used herein refers to the
number of
carbon atoms in a given group. Thus, a C1_6alkyl group contains from 1 to 6
carbon
atoms, a C3_6cycloalkyl group contains from 3 to 6 carbon atoms, and so on.
The term `C1_4alkyr as used herein as a group or part of a group represents a
straight or
branched chain saturated hydrocarbon radical having from 1 to 4 carbon atoms,
such as
methyl, ethyl, n-propyl, isopropyl, n-butyl, s-butyl, t-butyl and the like.
The term `C2_4alkyr as used herein as a group or part of a group represents a
straight or
branched chain saturated hydrocarbon radical having from 2 to 4 carbon atoms,
such as
ethyl, n-propyl, isopropyl, n-butyl, s-butyl, t-butyl and the like.
The term `C1_6alkyr as used herein as a group or part of a group represents a
straight or
branched chain saturated hydrocarbon radical having from 1 to 6 carbon atoms
such as
the groups defined for C1_4alkyl and n-pentyl, n-hexyl, 2-methylbutyl and the
like.
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The term `C3_6alkyr as used herein as a group or part of a group represents a
straight or
branched chain saturated hydrocarbon radical having from 3 to 6 carbon atoms
such as
n-propyl, isopropyl, n-butyl, s-butyl, t-butyl, n-pentyl, n-hexyl, 2-
methylbutyl and the
like.
The term `C3_5cycloalkyr as used herein as a group or part of a group defines
a saturated,
cyclic hydrocarbon radical having from 3 to 5 carbon atoms, such as
cyclopropyl,
cyclobutyl and cyclopentyl.
The term `spirobicyclic' as used herein as group or part of a group represents
cyclic
systems wherein two cycles are joined at a single atom. Examples of these
systems are
7- to 10-membered saturated spiroheterobicyclic systems containing one or two
N-atoms,
wherein one of the nitrogen atoms is always linked to the thienopyrimidinyl
heterocycle
in the compounds of Formula (I) as defined herein. Such spirocyclic systems
include,
but are not limited to systems resulting from the combination of e.g.
piperidine,
pyrrolidine, azetidine, and cyclobutane rings. Examples of such systems
include, but are
not limited to (a), (b), (c), (d), (e), (f) and (g) below and the like
0N' cysiN cNIN- -
O
N
N N N a i (d)
i a (a) i a (b) a , (c)
i
N g
a i (e)
-..., N
N a a i (g)
i (0
wherein a represents the position of linkage to the thienopyrimidinyl
heterocycle. The
skilled person will understand that in these particular examples, the options
for --L2-R3
defined herein when L2 is linked to a nitrogen atom of Ll, apply to examples
(a)-(f);
while the options for --L2-R3 defined herein when L2 is linked to a carbon
atom of Ll,
apply to example (g).
Examples of 7- to 10-membered saturated spirocarbobicyclic systems include,
but are
not limited to
0
0
, , ,
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====00 ====00 =====q0
.00 ___()0, - --%
,
and the like.
In general, whenever the term 'substituted' is used in the present invention,
it is meant,
unless otherwise indicated or clear from the context, to indicate that one or
more
hydrogens, in particular from 1 to 4 hydrogens, more in particular from 1 to 3
hydrogens,
preferably 1 or 2 hydrogens, more preferably 1 hydrogen, on the atom or
radical indicated
in the expression using 'substituted' are replaced with a selection from the
indicated
group, provided that the normal valency is not exceeded, and that the
substitution results
in a chemically stable compound, i.e. a compound that is sufficiently robust
to survive
isolation to a useful degree of purity from a reaction mixture.
Combinations of substituents and/or variables are permissible only if such
combinations
result in chemically stable compounds. 'Stable compound' is meant to indicate
a
compound that is sufficiently robust to survive isolation to a useful degree
of purity from
a reaction mixture.
The skilled person will understand that when an atom or radical is substituted
with 'a
substituent', it is meant that the atom or radical referred to is substituted
with one
substituent selected from the indicated group.
The skilled person will understand that the term 'optionally substituted'
means that the
atom or radical indicated in the expression using 'optionally substituted' may
or may not
be substituted (this means substituted or unsubstituted respectively).
When two or more substituents are present on a moiety they may, where possible
and
unless otherwise indicated or clear from the context, replace hydrogens on the
same atom
or they may replace hydrogen atoms on different atoms in the moiety.
It will be clear for the skilled person that, unless otherwise is indicated or
is clear from
the context, a substituent on a heterocyclyl group may replace any hydrogen
atom on a
ring carbon atom or on a ring heteroatom.
Within the context of this invention 'saturated' means 'fully saturated', if
not otherwise
specified.
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A 'non-aromatic group' embraces unsaturated ring systems without aromatic
character,
partially saturated and fully saturated carbocyclic and heterocyclic ring
systems. The
term 'partially saturated' refers to rings wherein the ring structure(s)
contain(s) at least
one multiple bond e.g. a C=C, N=C bond. The term 'fully saturated' refers to
rings where
there are no multiple bonds between ring atoms. Thus, a 'non-aromatic
heterocyclyl' is
a non-aromatic monocyclic or bicyclic system, unless otherwise specified,
having for
example, 3 to 12 ring members, more usually 5 to 10 ring members. Examples of
monocyclic groups are groups containing 4 to 7 ring members, more usually, 5
or 6 ring
members. Examples of bicyclic groups are those containing 8 to 12, more
usually 9 or
.. 10 ring members.
Non-limiting examples of monocyclic heterocyclyl systems containing at least
one
heteroatom selected from nitrogen, oxygen or sulfur (N, 0, S) include, but are
not limited
to 4- to 7-membered heterocyclyl systems such as azetidinyl, oxetanyl,
pyrrolidinyl,
tetrahydrofuranyl, piperidinyl, piperazinyl, pyranyl, dihydropyranyl,
tetrahydropyranyl,
.. morpholinyl, thiomorpholinyl, and tetrahydro-2H-thiopyranyl 1,1-dioxide, in
particular
azetidinyl, oxetanyl, pyrrolidinyl, tetrahydrofuranyl, piperidinyl,
piperazinyl, pyranyl,
dihydropyranyl, tetrahydropyranyl, morpholinyl, and thiomorpholiny. Non-
limiting
examples of bicyclic heterocyclyl systems containing at least one heteroatom
selected
from nitrogen, oxygen or sulfur (N, 0, S) include, but are not limited to
octahydro-1H-
N
al H
or 8
indolyl, indolinyl, " . Unless otherwise specified, each can
be bound to the remainder of the molecule of Formula (I) through any available
ring
carbon atom (C-linked) or nitrogen atom (N-linked), and may optionally be
substituted,
where possible, on carbon and/or nitrogen atoms according to the embodiments.
Examples of a C-linked 4- to 7-membered non-aromatic heterocyclyl containing
at least
.. one nitrogen atom include, but are not limited to, azetidinyl, pyrrolidinyl
and piperidinyl,
bound to the rest of the molecule through an available carbon atom.
The term `C-linked 4- to 6-membered heterocyclyl containing an oxygen atom' as
used
herein alone or as part of another group, defines a saturated, cyclic
hydrocarbon radical
containing an oxygen atom having from 4 to 6 ring members, such as oxetanyl,
tetrahydrofuranyl, and tetrahydropyranyl.
Whenever substituents are represented by chemical structure, `---' represents
the bond of
attachment to the remainder of the molecule of Formula (I).
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Lines (such as `---') drawn into ring systems indicate that the bond may be
attached to
any of the suitable ring atoms.
Het' and Het2 may be attached to the remainder of the molecule of Formula (I)
through
any available ring carbon or nitrogen atom as appropriate, if not otherwise
specified.
It will be clear that a saturated cyclic moiety may, where possible, have
substituents on
both carbon and N-atoms, unless otherwise is indicated or is clear from the
context.
It will be clear that when L2 is >S02, this is equivalent to L2 is -SO2-. It
will be clear
R4a R4b
'I
that when L2 is >CR4aR4b, this is equivalent to L is -c- . For example, in
compound 1, L2 is >CR4aR4b wherein both R4a and R4b are hydrogen.
4c 4d
R
¨C-
Similar, it will be clear that when L2 is >CR4cR4d, this is equivalent to L is
.
When any variable occurs more than one time in any constituent, each
definition is
independent.
When any variable occurs more than one time in any formula (e.g. Formula (I)),
each
definition is independent.
The term 'subject' as used herein, refers to an animal, preferably a mammal
(e.g. cat,
dog, primate or human), more preferably a human, who is or has been the object
of
treatment, observation or experiment.
The term 'therapeutically effective amount' as used herein, means that amount
of active
compound or pharmaceutical agent that elicits the biological or medicinal
response in a
tissue system, animal or human that is being sought by a researcher,
veterinarian,
medicinal doctor or other clinician, which includes alleviation or reversal of
the
symptoms of the disease or disorder being treated.
The term 'composition' is intended to encompass a product comprising the
specified
ingredients in the specified amounts, as well as any product which results,
directly or
indirectly, from combinations of the specified ingredients in the specified
amounts.
The term 'treatment', as used herein, is intended to refer to all processes
wherein there
may be a slowing, interrupting, arresting or stopping of the progression of a
disease, but
does not necessarily indicate a total elimination of all symptoms.
The term `compound(s) of the (present) invention' or `compound(s) according to
the
(present) invention' as used herein, is meant to include the compounds of
Formula (I)
and the pharmaceutically acceptable salts, and the solvates thereof.
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As used herein, any chemical formula with bonds shown only as solid lines and
not as
solid wedged or hashed wedged bonds, or otherwise indicated as having a
particular
configuration (e.g. R, S) around one or more atoms, contemplates each possible
stereoisomer, or mixture of two or more stereoisomers.
Hereinbefore and hereinafter, the term ' compound(s) of Formula (I)' is meant
to include
the tautomers thereof and the stereoisomeric forms thereof
The terms `stereoisomers', `stereoisomeric forms' or `stereochemically
isomeric forms'
hereinbefore or hereinafter are used interchangeably.
The invention includes all stereoisomers of the compounds of the invention
either as a
pure stereoisomer or as a mixture of two or more stereoisomers.
Enantiomers are stereoisomers that are non-superimposable mirror images of
each other.
A 1:1 mixture of a pair of enantiomers is a racemate or racemic mixture.
Atropisomers (or atropoisomers) are stereoisomers which have a particular
spatial
configuration, resulting from a restricted rotation about a single bond, due
to large steric
hindrance. All atropisomeric forms of the compounds of Formula (I) are
intended to be
included within the scope of the present invention.
Diastereomers (or diastereoisomers) are stereoisomers that are not
enantiomers, i.e. they
are not related as mirror images. If a compound contains a double bond, the
substituents
may be in the E or the Z configuration.
Substituents on bivalent cyclic saturated or partially saturated radicals may
have either
the cis- or trans-configuration; for example if a compound contains a
disubstituted
cycloalkyl group, the substituents may be in the cis or trans configuration.
Therefore, the invention includes enantiomers, atropisomers, diastereomers,
racemates,
E isomers, Z isomers, cis isomers, trans isomers and mixtures thereof,
whenever
chemically possible.
The meaning of all those terms, i.e. enantiomers, atropisomers, diastereomers,
racemates,
E isomers, Z isomers, cis isomers, trans isomers and mixtures thereof are
known to the
skilled person.
The absolute configuration is specified according to the Cahn-Ingold-Prelog
system. The
configuration at an asymmetric atom is specified by either R or S. Resolved
stereoisomers whose absolute configuration is not known can be designated by
(+) or
(-) depending on the direction in which they rotate plane polarized light. For
instance,
resolved enantiomers whose absolute configuration is not known can be
designated by
(+) or (-) depending on the direction in which they rotate plane polarized
light.
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When a specific stereoisomer is identified, this means that said stereoisomer
is
substantially free, i.e. associated with less than 50%, preferably less than
20%, more
preferably less than 10%, even more preferably less than 5%, in particular
less than 2%
and most preferably less than 1%, of the other stereoisomers. Thus, when a
compound
of Formula (I) is for instance specified as (R), this means that the compound
is
substantially free of the (5) isomer; when a compound of Formula (I) is for
instance
specified as E, this means that the compound is substantially free of the Z
isomer; when
a compound of Formula (I) is for instance specified as cis, this means that
the compound
is substantially free of the trans isomer.
Some of the compounds according to Formula (I) may also exist in their
tautomeric form.
Such forms in so far as they may exist, although not explicitly indicated in
the above
Formula (I) are intended to be included within the scope of the present
invention. It
follows that a single compound may exist in both stereoisomeric and tautomeric
form.
Pharmaceutically acceptable salts include acid addition salts and base
addition salts.
Such salts may be formed by conventional means, for example by reaction of a
free acid
or a free base form with one or more equivalents of an appropriate base or
acid, optionally
in a solvent, or in a medium in which the salt is insoluble, followed by
removal of said
solvent, or said medium, using standard techniques (e.g. in vacuo, by freeze-
drying or by
filtration). Salts may also be prepared by exchanging a counter-ion of a
compound of the
invention in the form of a salt with another counter-ion, for example using a
suitable ion
exchange resin.
The pharmaceutically acceptable salts as mentioned hereinabove or hereinafter
are meant
to comprise the therapeutically active non-toxic acid and base salt forms
which the
compounds of Formula (I) and solvates thereof, are able to form.
.. Appropriate acids comprise, for example, inorganic acids such as hydrohalic
acids, e.g.
hydrochloric or hydrobromic acid, sulfuric, nitric, phosphoric and the like
acids; or
organic acids such as, for example, acetic, propanoic, hydroxyacetic, lactic,
pyruvic,
oxalic (i.e. ethanedioic), malonic, succinic (i.e. butanedioic acid), maleic,
fumaric, malic,
tartaric, citric, methanesulfonic, ethanesulfonic, benzenesulfonic, p-
toluenesulfonic,
cyclamic, salicylic, p-aminosalicylic, pamoic and the like acids. Conversely
said salt
forms can be converted by treatment with an appropriate base into the free
base form.
The compounds of Formula (I) and solvates thereof containing an acidic proton
may also
be converted into their non-toxic metal or amine salt forms by treatment with
appropriate
organic and inorganic bases.
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Appropriate base salt forms comprise, for example, the ammonium salts, the
alkali and
earth alkaline metal salts, e.g. the lithium, sodium, potassium, cesium,
magnesium,
calcium salts and the like, salts with organic bases, e.g. primary, secondary
and tertiary
aliphatic and aromatic amines such as methylamine, ethylamine, propylamine,
isopropylamine, the four butylamine isomers, dimethylamine, diethylamine,
diethanolamine, dipropylamine, diisopropylamine, di-n-butylamine, pyrrolidine,
piperidine, morpholine, trimethylamine, triethylamine, tripropylamine,
quinuclidine,
pyridine, quinoline and isoquinoline; the benzathine, N-methyl-D-glucamine,
hydrabamine salts, and salts with amino acids such as, for example, arginine,
lysine and
the like. Conversely the salt form can be converted by treatment with acid
into the free
acid form.
The term solvate comprises the solvent addition forms as well as the salts
thereof, which
the compounds of Formula (I) are able to form. Examples of such solvent
addition forms
are e.g. hydrates, alcoholates and the like.
The compounds of the invention as prepared in the processes described below
may be
synthesized in the form of mixtures of enantiomers, in particular racemic
mixtures of
enantiomers, that can be separated from one another following art-known
resolution
procedures. A manner of separating the enantiomeric forms of the compounds of
Formula (I), and pharmaceutically acceptable salts, and solvates thereof,
involves liquid
chromatography using a chiral stationary phase. Said pure stereochemically
isomeric
forms may also be derived from the corresponding pure stereochemically
isomeric forms
of the appropriate starting materials, provided that the reaction occurs
stereospecifically.
Preferably if a specific stereoisomer is desired, said compound would be
synthesized by
stereospecific methods of preparation. These methods will advantageously
employ
enantiomerically pure starting materials.
The present invention also embraces isotopically-labeled compounds of the
present
invention which are identical to those recited herein, but for the fact that
one or more
atoms are replaced by an atom having an atomic mass or mass number different
from the
atomic mass or mass number usually found in nature (or the most abundant one
found in
nature).
All isotopes and isotopic mixtures of any particular atom or element as
specified herein
are contemplated within the scope of the compounds of the invention, either
naturally
occurring or synthetically produced, either with natural abundance or in an
isotopically
enriched form. Exemplary isotopes that can be incorporated into compounds of
the
invention include isotopes of hydrogen, carbon, nitrogen, oxygen, phosphorus,
sulfur,
fluorine, chlorine and iodine, such as 2H5 3H5 HC5 13c5 '4C, 13N5 1505 1705
1805 32P5 33P5
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1 75131.5 76131.5 77Br and 82Br. Preferably, the radioactive
isotope is selected from the group of2H,3H, "C and 18F. More preferably, the
radioactive
isotope is 2H. In particular, deuterated compounds are intended to be included
within the
scope of the present invention.
Certain isotopically-labeled compounds of the present invention (e.g., those
labeled with
3H and 14C) may be useful for example in substrate tissue distribution assays.
Tritiated
(3H) and carbon-14 (14C) isotopes are useful for their ease ofpreparation and
detectability.
Further, substitution with heavier isotopes such as deuterium (i.e., 2H may
afford certain
therapeutic advantages resulting from greater metabolic stability (e.g.,
increased in vivo
half-life or reduced dosage requirements) and hence may be preferred in some
circumstances. Thus, in a particular embodiment of the present invention, R2
is selected
from hydrogen or deuterium, in particular deuterium. In another embodiment, L2
can be
>C(2H)2. Positron emitting isotopes such as 150, 13N5 "C and 18F are useful
for positron
emission tomography (PET) studies. PET imaging in cancer finds utility in
helping
locate and identify tumours, stage the disease and determine suitable
treatment. Human
cancer cells overexpress many receptors or proteins that are potential disease-
specific
molecular targets. Radiolabelled tracers that bind with high affinity and
specificity to
such receptors or proteins on tumour cells have great potential for diagnostic
imaging
and targeted radionuclide therapy (Charron, Carlie L. et al. Tetrahedron Lett.
2016,
57(37), 4119-4127). Additionally, target-specific PET radiotracers may be used
as
biomarkers to examine and evaluate pathology, by for example, measuring target
expression and treatment response (Austin R. et al. Cancer Letters (2016),
doi:
10.1016/j.canlet.2016.05.008).
The present invention relates in particular to compounds of Formula (I) and
the
pharmaceutically acceptable salts, and the solvates thereof, or any subgroup
thereof as
mentioned in any of the embodiments, wherein also the following compound and
pharmaceutically acceptable addition salts, and solvates thereof, are
excluded:
OH
&
N N
N 0 NC)
F H
__________ / I N
/
S----N
FiF
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The present invention relates in particular to compounds of Formula (I) and
the
pharmaceutically acceptable salts, and the solvates thereof, or any subgroup
thereof as
mentioned in any of the embodiments, wherein the intermediates and compounds
described in W02017/112768, in as far as they are covered by the present
invention, are
excluded.
The present invention relates in particular to compounds of Formula (I) as
defined herein,
and the tautomers and the stereoisomeric forms thereof, wherein
Rl is CF3;
R2 is selected from the group consisting of hydrogen and CH3;
Ll represents a 7- to 10-membered saturated spiroheterobicyclic system
containing one
or two N-atoms provided that it is N-linked to the thienopyrimidinyl
heterocycle; and
--L2-R3 is selected from (a), (b), (c), (d), (f) or (g), wherein
(a) L2 is selected from the group consisting of >CR4aR4b, and -CHR4aCHR5-;
wherein
L2 is linked to a nitrogen atom of Ll; R4a is selected from the group
consisting of hydrogen; -C(=0)NR7aR7b; Ci_4alkyl optionally substituted
with a substituent selected from the group consisting of -0R8, and -
NR9aR9b; and C-linked 4- to 7-membered non-aromatic heterocyclyl
containing at least one nitrogen atom;
R5 is selected from the group consisting of hydrogen; -0R6; and Ci_4a1ky1;
R4b is selected from the group consisting of hydrogen and methyl; or
R4a and R4b together with the carbon atom to which they are attached form a C3-
5cyc1oa1ky1 or a C-linked 4- to 6-membered heterocyclyl containing an oxygen
atom; wherein
R6, R7a, R7b, R8, R9a and R9b are each independently selected from the group
consisting of hydrogen; and C2_4alkyl substituted with a substituent selected
from
the group consisting of -OR" and -NR10aRlOb; wherein
R10a5 RlOb and R"
are each independently selected from the group consisting of
hydrogen; and Ci_4a1ky1; and
R3 is selected from the group consisting of Ar, Het', Het2, and a 7- to 10-
membered saturated spirocarbobicyclic system; or
(b) L2 is >CR4cR4d, wherein R4c and Wid are hydrogen; and
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R12a
I
,Ge......R12b
-- 1 R3 is R12c ; wherein R', R12b, and R12c are C1_6alkyl 1; or
(c) --L2-R3 is Ci_6alkyl optionally substituted with one, two or three fluoro
substituents;
Or
(d) L2 is 0 and R3 is -CH2-Ar; or
R18a
R18Ic )........")
N
I 5 (f) --L2-R3 is R 18 5 wherein
R" is selected from the group consisting of hydrogen; and C1_4alkyl;
R18a is selected from the group consisting of hydrogen, and fluoro;
R18b is selected from the group consisting of fluoro, -0C1_4alkyl, and
C1_4alkyl
optionally substituted with 1, 2 or 3 fluoro substituents; or
R18a and R18b are bound to the same carbon atom and together form a
C3_5cycloalkyl; or
CcN..)
N-
---...
(g) --L2-R3 is ; and wherein
Ar is phenyl which may be optionally substituted with one, two, or three
substituents
each independently selected from the group consisting of halo,
-0R24, and C1_4alkyl optionally substituted with -0R26;
Het' is a monocyclic heteroaryl selected from the group consisting of pyridyl,
4-, 5- or
6-pyrimidinyl, pyrazinyl, pyridazinyl, pyrrolyl, pyrazolyl, imidazolyl, 4- or
5-thiazolyl,
isothiazolyl, and isoxazolyl; or a bicyclic heteroaryl selected from the group
consisting
of indolyl, imidazopyridinyl; each of which may be optionally substituted with
one, two,
or three substituents each independently selected from the group consisting of
halo, -CN,
-0R24, and C1_4alkyl optionally substituted with a substituent selected from
the group
consisting of -CN, -0R26, and -NR27aR271); and
Het2 is a non-aromatic heterocyclyl optionally substituted with one, two, or
three
substituents each independently selected from the group consisting of halo, -
CN, and
Ci_4alkyl optionally substituted with -0R26;
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wherein
R245 R265 R27a5 and R27b are each independently selected from the group
consisting of
hydrogen; Ci_4a1ky1; and C2_4alkyl substituted with ¨NR28aR28b; wherein
.. R28a and R28b are hydrogen;
and the pharmaceutically acceptable salts and the solvates thereof.
The present invention relates in particular to compounds of Formula (I) as
defined herein,
and the tautomers and the stereoisomeric forms thereof, wherein
(a) L2 is selected from the group consisting of >S02, >CR4aR4b5 and -CHR4aCHR5-
;
wherein
(i) when L2 is linked to a carbon atom of Ll, then R4a and R5 are each
independently selected from the group consisting of hydrogen; -0R6;
-NR7aR7b; -C(=0)NR7aR7b; Ci_4alkyl optionally substituted with a
substituent selected from the group consisting of fluoro, -CN, -OR% and
-NR9aR9b; and C-linked 4- to 7-membered non-aromatic heterocyclyl
containing at least one nitrogen, oxygen or sulfur atom;
(ii) when L2 is linked to a nitrogen atom of Ll, then R4a is selected
from the
group consisting of hydrogen; -C(=0)NR7aR7b; Ci_4alkyl optionally
substituted with a substituent selected from the group consisting of fluoro,
-CN, -01e, and -NR9aR9b; and C-linked 4- to 7-membered non-aromatic
heterocyclyl containing at least one nitrogen, oxygen or sulfur atom;
R5 is selected from the group consisting of hydrogen; -0R6; -NR7aR7b;
-C(=0)NR7aR7b; Ci_4a1ky1 optionally substituted with a substituent selected
from the group consisting of fluoro, -CN, -OR% and -NR9aR9b; and C-linked
4- to 7-membered non-aromatic heterocyclyl containing at least one
nitrogen, oxygen or sulfur atom;
R4b is selected from the group consisting of hydrogen and methyl; or >CR4aR4b
form a >C3_5cycloalkanediy1 or a >C-linked 4- to 6-membered heterocyclediyl
containing an oxygen atom; wherein
R6, R7a, R7b, R8, R9a and R9b are each independently selected from the group
consisting of hydrogen; Ci_4a1ky1 optionally substituted with a substituent
selected from the group consisting of fluoro, -CN, and -C(=0)NR10aRlOb; and C2-
4alkyl substituted with a substituent selected from the group consisting of
-OR" and -NR10aRlOb; wherein
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Rioa, Riob and R"
are each independently selected from the group consisting of
hydrogen; and C1_4alkyl; and
R3 is selected from the group consisting of Ar, Het', Het2, and a 7- to 10-
membered saturated spirocarbobicyclic system; or
(b) L2 is selected from the group consisting of >cR4cR4d and -CHR4cCHR5a-,
wherein
R4c5 R4a5 and R5a are each independently selected from the group consisting of
hydrogen and C1_4alkyl; and
R
12a R 12a
I I
.si Ri2b ,Ge_R12b
-- '12c -- '12c
R3 is selected from the group consisting of R and R ;
wherein Rua, R12b5 and R12c are each independently selected from the group
consisting of C1_6alkyl optionally substituted with a ¨OH or a ¨NH2
substituent;
Or
(C) --L2-R3 is C1_6alkyl optionally substituted with one, two or three fluoro
substituents;
Or
(d) L2 is 0 and R3 is selected from the group consisting of Ar, He-0; -CH2-Ar,
-CH2-Het', and -CH2-(a 7- to 10-membered saturated spirocarbobicyclic system);
when L2 is linked to a carbon atom of Ll; or
0
N
118 44
(e) --L2-R3 is selected from
the group consisting of R 5 wherein
R" is hydrogen; or
cr:1:/1,
N-
---...õ S
..---
N
(0 --L2-R3 is Or ; and wherein
Ar is phenyl optionally substituted with one, two, or three substituents each
independently selected from the group consisting of halo, -CN, and C1_4alkyl
optionally
substituted with a substituent selected from the group consisting of fluoro, -
CN, -0R26, -
NR27aR27b, and ¨C(=0)NR27aR27b;
Het' is a monocyclic heteroaryl selected from the group consisting of pyridyl,
4-, 5- or
6-pyrimidinyl, pyrazinyl, pyridazinyl, furanyl, thienyl, pyrrolyl, pyrazolyl,
imidazolyl,
4- or 5-thiazolyl, isothiazolyl, thiadiazolyl, and isoxazolyl; or a bicyclic
heteroaryl
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selected from imidazopyridinyl, in particular imidazo[1,2-a]pyridinyl; each of
which
may be optionally substituted with one, two, or three substituents each
independently
selected from the group consisting of halo, -CN, and C1_4alkyl optionally
substituted with
a substituent selected from the group consisting of fluoro, -CN, -0R26, -
NR27aR27b5 and
¨C(=0)NR27aR27b; and
Het2 is a non-aromatic heterocyclyl selected from azetidinyl, pyrrolidinyl and
piperidinyl;
wherein
R265 R27a5 and R27b are each independently selected from the group consisting
of hydrogen
and Ci_4alkyl;
and the pharmaceutically acceptable salts and the solvates thereof.
The present invention relates in particular to compounds of Formula (I) as
defined herein,
and the tautomers and the stereoisomeric forms thereof, wherein
Rl is selected from the group consisting of CH3, CH2F, CHF2, and CF3;
R2 is selected from the group consisting of hydrogen and CH3;
Ll represents a 7- to 10-membered saturated spiroheterobicyclic system
containing one
or two N-atoms provided that it is N-linked to the thienopyrimidinyl
heterocycle; and
--L2-R3 is selected from (a), (b), (d), (e), or (f), wherein
(a) L2 is selected from the group consisting of >S02, >CR4aR4b, and -CHR4aCHR5-
;
wherein
(0 when L2 is linked to a carbon atom of Ll, then R4a and R5 are each
independently selected from the group consisting of hydrogen; -0R6;
-NR7aR7b; -C(=0)NR7aR7b; Ci_4alkyl optionally substituted with a
substituent selected from the group consisting of fluoro, -CN, -OR% and
-NR9aR9b; and C-linked 4- to 7-membered non-aromatic heterocyclyl
containing at least one nitrogen, oxygen or sulfur atom;
(ii) when L2 is linked to a nitrogen atom of Ll, then R4a is
selected from the
group consisting of -C(=0)NR7aR7b; Ci_4alkyl optionally substituted with
a substituent selected from the group consisting of fluoro, -CN, -01e, and
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-NR9aR9b; and C-linked 4- to 7-membered non-aromatic heterocyclyl
containing at least one nitrogen, oxygen or sulfur atom;
R5 is selected from the group consisting of hydrogen; -0R6; -NR7aR7b;
-C(=0)NR7aR7b; Ci_4alkyl optionally substituted with a substituent
selected from the group consisting of fluoro, -CN, -0R8, and
-NR9aR9b; and C-linked 4- to 7-membered non-aromatic heterocyclyl
containing at least one nitrogen, oxygen or sulfur atom;
R4b is selected from the group consisting of hydrogen and methyl; or R4a and
R4b
together with the carbon atom to which they are attached form a C3_5cycloalkyl
or a C-linked 4- to 6-membered heterocyclyl containing an oxygen atom; wherein
R6, R7a, R7b, R8, R9a and R9b are each independently selected from the group
consisting of hydrogen; Ci_4a1ky1 optionally substituted with a substituent
selected from the group consisting of fluoro, -CN, and -C(=0)NR10aRlOb; and C2-
4alkyl substituted with a substituent selected from the group consisting of
-0R11 and -NR10aRlOb; wherein
Rioa, RlOb and R"
are each independently selected from the group consisting of
hydrogen; Ci_4a1ky1; and C-linked 4- to 7-membered non-aromatic heterocyclyl
containing at least one nitrogen, oxygen or sulfur atom; and
R3 is selected from the group consisting of Ar, Het', Het2, and a 7- to 10-
membered saturated spirocarbobicyclic system; or
(b) L2 is selected from the group consisting of >cR4cR4d and -CHR4cCHR5a-,
wherein
R4c5 R4a5 and R5a are each independently selected from the group consisting of
hydrogen and Ci_4a1ky1; and
R R
12a 12a
I I
.si Ri2b ,Ge _Ri 2b
-- '12c
R3 is selected from the group consisting of R and R ;
wherein R12a, Rim, and R12c are each independently selected from the group
consisting of Ci_6a1ky1 optionally substituted with a ¨OH or a ¨NH2
substituent;
and -0Ci_6a1ky1; or
(d) L2 is 0 and R3 is selected from the group consisting of C3_6alkyl
optionally
substituted with one, two or three fluoro substituents; Ar; Het'; Het2; a 7-
to 10-
membered saturated spirocarbobicyclic system; -CH2-Ar; -CH2-Het'; -CH2-Het2;
and -CH2-(a 7-to 10-membered saturated spirocarbobicyclic system); when L2 is
linked to a carbon atom of L1; or
(e) --L2-R3 is ¨0-CHR5-R3 when L2 is linked to a carbon atom of L1, wherein
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R5 is selected from the group consisting of -C(=0)NR13aRl3b;
Ci_4a1ky1 optionally substituted with a substituent selected from the group
consisting of fluoro, -OR", and -NR15aRl5b; and C-linked 4-to 7-membered non-
aromatic heterocyclyl containing at least one nitrogen, oxygen or sulfur atom;
wherein
R13a, R13b5 R145 R15a and Risb are each independently selected from the group
consisting of hydrogen; Ci_4a1ky1 optionally substituted with a substituent
selected from the group consisting of fluoro and -C(=0)NR16aRl6b; and
C2_4alkyl substituted with a substituent selected from the group consisting of
-OR' and -NR16aRl6b; wherein
R16a5 R16b and K-17
are each independently selected from the group consisting of
hydrogen; Ci_zialkyl; and C-linked 4- to 7-membered non-aromatic heterocyclyl
containing at least one nitrogen, oxygen or sulfur atom; and
R3 is selected from the group consisting of hydrogen; Ci_4a1ky1 optionally
substituted with one, two, or three fluoro substituents; -CN; Ar, Het'; Het2;
and a
7- to 10-membered saturated spirocarbobicyclic system; or
R18a
R181::. )........0
N
I (0 --L2-R3 is R18 5 wherein
R" is selected from the group consisting of hydrogen; Ci_4a1ky1 optionally
substituted with a fluoro or a ¨CN substituent; and C2_4alkyl substituted with
a
substituent selected from the group consisting of ¨OR' and ¨NR2 aR
20b; wherein
V, R20a and R2 b are each independently selected from the group consisting of
hydrogen; Ci_4a1ky1 optionally substituted with a substituent selected from
the
group consisting of fluoro, -CN, and -C(=0)NR2laR21b; C2_4alkyl substituted
with
a substituent selected from the group consisting of ¨0R22 and _NR2iaR21b; and
c _
linked 4- to 7-membered non-aromatic heterocyclyl containing at least one
nitrogen, oxygen or sulfur atom; wherein
R21a5 R2lb and K-.22
are each independently selected from the group consisting of
hydrogen and Ci_zialkyl; and
R"a is selected from the group consisting of hydrogen, fluoro and Ci_zialkyl;
Ri" is selected from the group consisting of fluoro, -0Ci_4a1ky1, and
Ci_4a1ky1
optionally substituted with 1, 2 or 3 fluoro substituents; or
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R18a and Ri8b are bound to the same carbon atom and together form a
C3_5cycloalkyl or a C-linked 4- to 6-membered heterocyclyl containing an
oxygen
atom;
and wherein
Ar is phenyl or naphthyl, each of which may be optionally substituted with
one, two, or
three substituents each independently selected from the group consisting of
halo, -CN,
-OR', -NR25aR25b5 and C1_4alkyl optionally substituted with a substituent
selected from
the group consisting of fluoro, -CN, -0R26, -NR27aR27b, and ¨C(=0)NR27aR27b;
Het' is a monocyclic heteroaryl selected from the group consisting of pyridyl,
4-, 5- or
6-pyrimidinyl, pyrazinyl, pyridazinyl, furanyl, thienyl, pyrrolyl, pyrazolyl,
imidazolyl,
4- or 5-thiazolyl, isothiazolyl, thiadiazolyl, and isoxazolyl; or a bicyclic
heteroaryl
selected from the group consisting of imidazothiazolyl, imidazoimidazolyl,
benzofuranyl, benzothiophenyl, benzimidazolyl, benzoxazo lyl, isobenzoxazo
lyl,
benziso xazo lyl, benzothiazo lyl, benzisothiazo lyl, isobenzofuranyl, indo
lyl, iso indolyl,
indolizinyl, indolinyl, isoindolinyl, indazo lyl, pyrazolopyridinyl,
pyrazolopyrimidinyl,
imidazopyridinyl, imidazopyrazinyl, imidazopyridazinyl; each of which may be
optionally substituted with one, two, or three substituents each independently
selected
5 _NR25aR25b5
from the group consisting of halo, -CN, _0R24 and
C1_4alkyl optionally
substituted with a substituent selected from the group consisting of fluoro, -
CN, -0R26, -
NR27aR27b5 and ¨C(=0)NR27aR27b; and
Het2 is a non-aromatic heterocyclyl optionally substituted with one, two, or
three
substituents each independently selected from the group consisting of halo, -
CN, -OR',
_NR25 aR2 5 b 5 and C1_4alkyl optionally substituted with a substituent
selected from the
group consisting of fluoro, -CN, -0R26, -NR27aR27b, and ¨C(=0)NR27aR27b;
wherein
R24, R25a, R25b, R26, R27a, and R27b are each independently selected from the
group
consisting of hydrogen; C1_4a1ky1 optionally substituted with a substituent
selected from
the group consisting of fluoro and -C(=0)NR28aR28b; and C2_4alkyl substituted
with a
substituent selected from the group consisting of -0R29 and ¨NR28aR28b;
wherein
R28a, R28b and K-29
are each independently selected from the group consisting of hydrogen;
C1_4alkyl; and C-linked 4- to 7-membered non-aromatic heterocyclyl containing
at least
one nitrogen, oxygen or sulfur atom;
and the pharmaceutically acceptable salts and the solvates thereof.
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The present invention relates in particular to compounds of Formula (I) as
defined herein,
and the tautomers and the stereoisomeric forms thereof, wherein
R1 is CF3;
(a) L2 is >cg4ag4b; wherein
R4a is selected from the group consisting of hydrogen; -C(=0)NR7aR7b;
Ci_4a1ky1; and C-linked 4- to 7-membered non-aromatic heterocyclyl containing
at least one nitrogen, oxygen or sulfur atom; and
R4b is selected from the group consisting of hydrogen and methyl; wherein
R7a and R7b are each independently selected from the group consisting of
hydrogen; Ci_4a1ky1; and C2_4alkyl substituted with a substituent selected
from the
group consisting of-OR'1 and -NR10aRlOb; wherein
gioa, glob and R"
are each independently selected from the group consisting of
hydrogen and Ci_4a1ky1; and
R3 is selected from the group consisting of Ar, Het', Het2, and a 7- to 10-
membered saturated spirocarbobicyclic system; or
(b) L2 is >cg4cg4d5 wherein R4c and Wid are each independently selected
from the
group consisting of hydrogen and Ci_4a1ky1; and
R12a
R12a
I I
.Sj R12b ,GeRi 2b
-- '12c
R3 is selected from the group consisting of R and R ;
wherein R12a, gin, and R12c are each independently selected from the group
consisting of Ci_6a1ky1 optionally substituted with a ¨NH2 substituent; or
(c) --L2-R3 is Ci_6a1ky1 optionally substituted with one, two or three
fluoro
substituents; or
(d) L2 is 0 and R3 is selected from the group consisting of Ar, Het', -CH2-
Ar,
-CH2-Het', and -CH2-(a 7-to 10-membered saturated spirocarbobicyclic system);
when L2 is linked to a carbon atom of L1; or
'0.........0
N
I 18 44
(e) --L2-R3 is selected from the group consisting of R 5 wherein
R18 is hydrogen; or
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Cri:1:/1µ
N-
---.... S
-. -'
N
(0 --L2-R3 is or ; and wherein
Ar is phenyl optionally substituted with a halo substituent;
Het' is a monocyclic heteroaryl selected from the group consisting of pyridyl,
4-, 5- or
6-pyrimidinyl, pyrazinyl, pyridazinyl, pyrrolyl, pyrazolyl, imidazolyl, and 4-
or 5-
thiazolyl; or a bicyclic heteroaryl selected from imidazopyridinyl, in
particular
imidazo[1,2-a]pyridinyl; each of which may be optionally substituted with one,
two, or
three substituents each independently selected from the group consisting of
halo and
C1_4alkyl optionally substituted with a substituent selected from the group
consisting of
fluoro, -CN, -0R26, -NR27aR27b5 and ¨C(=0)NR27aR27b; and
Het2 is a non-aromatic heterocyclyl selected from azetidinyl, pyrrolidinyl and
piperidinyl;
wherein
R265 R27a5 and R27b are each independently selected from the group consisting
of hydrogen
and Ci_4alkyl;
and the pharmaceutically acceptable salts and the solvates thereof.
The present invention relates in particular to compounds of Formula (I) as
defined herein,
and the tautomers and the stereoisomeric forms thereof, wherein
Rl is CF3;
1_,1 represents a N-linked 7- to 10-membered saturated spiroheterobicyclic
system
containing one or two N-atoms selected from the group consisting of (a), (b),
(c), (d), (e),
(f) and (g)
i
'
O
i
0N ciNil yiN" -
N N N N
i a (a) i a (b) a , (c) a i (d)
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,
Q i
g
--..,
N N N
a i (e) a i (f) a i (g)
wherein a represents the position of linkage to the thienopyrimidinyl
heterocycle;
(a) L2 is >CH2; and R3 is selected from the group consisting of Ar, Het',
and a 7- to
10-membered saturated spirocarbobicyclic system; or
(b) --L2-R3 is C1_6alkyl optionally substituted with one, two or three
fluoro
substituents; and wherein
Ar is phenyl optionally substituted with a halo substituent; and
Het' is a monocyclic heteroaryl selected from the group consisting of 4-, 5-
or 6-
pyrimidinyl, pyrazinyl, pyridazinyl, pyrrolyl, pyrazolyl, imidazolyl, and 4-
or 5-
thiazolyl; or a bicyclic heteroaryl selected from imidazopyridinyl, in
particular
imidazo[1,2-a]pyridinyl; each of which may be optionally substituted with one
or two
sub stituents each independently selected from the group consisting of halo
and Ci_4alkyl;
and the pharmaceutically acceptable salts and the solvates thereof.
The present invention relates in particular to compounds of Formula (I) as
defined herein,
and the tautomers and the stereoisomeric forms thereof, wherein
Rl is CF3;
R2 is hydrogen;
Ll represents a N-linked 7- to 10-membered saturated spiroheterobicyclic
system
containing one or two N-atoms selected from the group consisting of (a), (b),
(c), (d), (e),
(f) and (g)
i
0N ciNil yiN" -
O
N N N N
i a (a) i a (b) a , (c) a i (d)
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,
Q i
g
--..,
N N N
a i (e) a i (0 a i (g)
wherein a represents the position of linkage to the thienopyrimidinyl
heterocycle;
(a) L2 is >CH2; and R3 is selected from the group consisting of Ar, Het',
and a 7- to
10-membered saturated spirocarbobicyclic system; or
(b) --L2-R3 is C1_6alkyl optionally substituted with one, two or three
fluoro
substituents; and wherein
Ar is phenyl optionally substituted with a halo substituent; and
Het' is a monocyclic heteroaryl selected from the group consisting of 4-, 5-
or 6-
.. pyrimidinyl, pyrazinyl, pyridazinyl, pyrrolyl, pyrazolyl, and imidazolyl;
or a bicyclic
heteroaryl selected from imidazopyridinyl, in particular imidazo[1,2-
a]pyridinyl; each of
which may be optionally substituted with one or two substituents each
independently
selected from the group consisting of halo and C1_4alkyl;
and the pharmaceutically acceptable salts and the solvates thereof
Another embodiment of the present invention relates to those compounds of
Formula (I)
and the pharmaceutically acceptable salts, and the solvates thereof, or any
subgroup
thereof as mentioned in any of the other embodiments wherein one or more of
the
following restrictions apply:
(a) Rl is CF3;
(b) R2 is hydrogen;
(c) Ll is a N-linked 7- to 10-membered saturated spiroheterobicyclic system
containing
one or two N-atoms selected from the group consisting of (a), (b), (c), (d),
(e), (f) and (g)
as defined herein;
(d) Ll is a N-linked 7- to 10-membered saturated spiroheterobicyclic system
containing
one or two N-atoms selected from the group consisting of (a), (b), (c), (d),
(e), and (f) as
defined herein;
(e) Ll is a N-linked 7- to 10-membered saturated spiroheterobicyclic system
containing
one or two N-atoms selected from the group consisting of (c) and (e);
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(f) L2 is >CH2;
(g) L2 is >CH2; and R3 is selected from the group consisting of Ar, Het', and
a 7- to 10-
membered saturated spirocarbobicyclic system;
'0.........0
N
118 44
(h) --L2-R3 is selected from the group consisting of R 5
F3C
N ,
, N N
1 ' , I ,
, 1
R18 R18 R18
5 5
¨0 F
N N N
1
F 18
RI18 R118
R
5 5 5
F----.4
),.......,0 Ct...,0
1 ' , 1 ,
,
R18
, and R18 ; wherein
5 R" is selected from the group consisting of hydrogen; Ci_4a1ky1
optionally
substituted with a fluoro or ¨CN substituent; and C2_4alkyl substituted with a
substituent selected from the group consisting of ¨OR' and ¨NR2 aR
20b; wherein
R19, R20a and R2 b are each independently selected from the group consisting
of
hydrogen; Ci_4a1ky1 optionally substituted with a substituent selected from
the
group consisting of fluoro, -CN, and -C(=0)NR2laR21b; C2_4alkyl substituted
with
a substituent selected from the group consisting of ¨0R22 and _NR2iaR21b; and
C-
linked 4- to 7-membered non-aromatic heterocyclyl containing at least one
nitrogen, oxygen or sulfur atom; wherein
R21a, R2lb and K-.22
are each independently selected from the group consisting of
hydrogen and Ci_4a1ky1;
(i) Ar is phenyl optionally substituted with one or two independently selected
halo
substituents;
(j) Ar is phenyl optionally substituted with one halo substituent;
(k) Ar is phenyl;
(1) Het' is a monocyclic heteroaryl selected from the group consisting of
pyrazolyl,
imidazolyl, pyrrolyl, 4- or 5-thiazolyl, pyridyl, pyridazinyl, 4-, 5- or 6-
pyrimidinyl, and
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pyrazinyl; or is a bicyclic heteroaryl selected from imidazopyridinyl, in
particular
imidazo[1,2-a]pyridinyl; each of which may be optionally substituted with one
or two
substituents each independently selected from the group consisting of halo and
C1_4a1kyl
optionally substituted with a substituent selected from the group consisting
of fluoro, -
CN, -0R25, -NR261R26b, and ¨C(=0)NR261R26b; wherein R25, R26a5 and R26b are
each
independently selected from the group consisting of hydrogen and C1_4alkyl;
(m) Het' is a monocyclic heteroaryl selected from the group consisting of
pyrazolyl,
imidazolyl, pyrrolyl, 4- or 5-thiazolyl, pyridyl, pyridazinyl, 4-, 5- or 6-
pyrimidinyl, and
pyrazinyl; or is a bicyclic heteroaryl selected from imidazopyridinyl, in
particular
imidazo[1,2-a]pyridinyl; each of which may be optionally substituted with one
or two
substituents each independently selected from the group consisting of halo and
C1_4a1kyl
optionally substituted with a substituent selected from the group consisting
of -CN, -
OR25, -NR261R26b5 and ¨C(=0)NR261R26b; wherein R25, R26a5 and R26b are each
independently selected from the group consisting of hydrogen and C1_4alkyl;
(n) Het' is a monocyclic heteroaryl selected from the group consisting of
pyrazolyl,
imidazolyl, pyrrolyl, pyridazinyl, 4-, 5- or 6-pyrimidinyl, and pyrazinyl; or
is a bicyclic
heteroaryl selected from imidazopyridinyl, in particular imidazo[1,2-
a]pyridinyl; each of
which may be optionally substituted with one or two substituents each
independently
selected from the group consisting of halo and C1_4a1kyl;
(o) Het' is a monocyclic heteroaryl selected from the group consisting of
pyridazinyl, 4-
5- or 6-pyrimidinyl, and pyrazinyl, each of which may be optionally
substituted with a
halo substituent;
(p) Het' is a bicyclic heteroaryl selected from imidazopyridinyl, in
particular
imidazo[1,2-a]pyridiny1-6-y1 or imidazo[1,2-a]pyridiny1-2-y1;
(q) 7- to 10-membered saturated spirocarbobicyclic system is in particular
=
In an embodiment, the present invention relates to those compounds of Formula
(I) and
the pharmaceutically acceptable salts, and the solvates thereof, or any
subgroup thereof
as mentioned in any of the other embodiments, wherein RI is CF3.
In an embodiment, the present invention relates to those compounds of Formula
(I) and
the pharmaceutically acceptable salts, and the solvates thereof, or any
subgroup thereof
as mentioned in any of the other embodiments, wherein RI is CF3, and wherein
R2 is
hydrogen.
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In an embodiment, the present invention relates to those compounds of Formula
(I) and
the pharmaceutically acceptable salts, and the solvates thereof, or any
subgroup thereof
as mentioned in any of the other embodiments, wherein Ar is phenyl optionally
substituted according to any of the other embodiments.
In an embodiment, the present invention relates to those compounds of Formula
(I) and
the pharmaceutically acceptable salts, and the solvates thereof, or any
subgroup thereof
as mentioned in any of the other embodiments, wherein L2 is linked to a carbon
atom of
L'.
In an embodiment, the present invention relates to those compounds of Formula
(I) and
the pharmaceutically acceptable salts, and the solvates thereof, or any
subgroup thereof
as mentioned in any of the other embodiments, wherein --L2-IV is (a).
In an embodiment, the present invention relates to those compounds of Formula
(I) and
the pharmaceutically acceptable salts, and the solvates thereof, or any
subgroup thereof
as mentioned in any of the other embodiments, wherein --L2-IV is (b).
In an embodiment, the present invention relates to those compounds of Formula
(I) and
the pharmaceutically acceptable salts, and the solvates thereof, or any
subgroup thereof
as mentioned in any of the other embodiments, wherein --L2-IV is (c).
In an embodiment, the present invention relates to those compounds of Formula
(I) and
the pharmaceutically acceptable salts, and the solvates thereof, or any
subgroup thereof
as mentioned in any of the other embodiments, wherein --L2-IV is (d).
In an embodiment, the present invention relates to those compounds of Formula
(I) and
the pharmaceutically acceptable salts, and the solvates thereof, or any
subgroup thereof
as mentioned in any of the other embodiments, wherein --L2-IV is (e).
In an embodiment, the present invention relates to those compounds of Formula
(I) and
the pharmaceutically acceptable salts, and the solvates thereof, or any
subgroup thereof
as mentioned in any of the other embodiments, wherein --L2-IV is (f).
In an embodiment, the present invention relates to those compounds of Formula
(I) and
the pharmaceutically acceptable salts, and the solvates thereof, or any
subgroup thereof
as mentioned in any of the other embodiments, wherein --L2-IV is (g).
In an embodiment, the present invention relates to those compounds of Formula
(I) and
the pharmaceutically acceptable salts, and the solvates thereof, or any
subgroup thereof
as mentioned in any of the other embodiments, wherein --L2-IV is (a); (b),
(d), (e) or (f);
and R4a is other than hydrogen.In an embodiment, the present invention relates
to those
compounds of Formula (I) and the pharmaceutically acceptable salts, and the
solvates
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thereof, or any subgroup thereof as mentioned in any of the other embodiments,
wherein
--L2-R3 is (a) or (f); and R4a is other than hydrogen.In an embodiment, the
present
invention relates to those compounds of Formula (I) and the pharmaceutically
acceptable
salts, and the solvates thereof, or any subgroup thereof as mentioned in any
of the other
embodiments, wherein --L2-R3 is (a); and R4a is other than hydrogen.
In an embodiment, the present invention relates to those compounds of Formula
(I) and
the pharmaceutically acceptable salts, and the solvates thereof, or any
subgroup thereof
as mentioned in any of the other embodiments, wherein --L2-R3 is (a); and when
L2 is
linked to a nitrogen atom of L' then R4a is other than hydrogen.
In an embodiment, the present invention relates to those compounds of Formula
(I) and
the pharmaceutically acceptable salts, and the solvates thereof, or any
subgroup thereof
as mentioned in any of the other embodiments, wherein --L2-R3 is (a) or (f);
and when
L2 is linked to a nitrogen atom of LI then R4a is other than hydrogen.
In an embodiment, the present invention relates to those compounds of Formula
(I) and
the pharmaceutically acceptable salts, and the solvates thereof, or any
subgroup thereof
as mentioned in any of the other embodiments, wherein --L2-R3 is (a), (b),
(d), (e) or (f);
and when L2 is linked to a nitrogen atom of L' then R4a is other than
hydrogen.
In an embodiment, the present invention relates to those compounds of Formula
(I) and
the pharmaceutically acceptable salts, and the solvates thereof, or any
subgroup thereof
as mentioned in any of the other embodiments, wherein LI represents
11
=
In an embodiment, the present invention relates to those compounds of Formula
(I) and
the pharmaceutically acceptable salts, and the solvates thereof, or any
subgroup thereof
as mentioned in any of the other embodiments, wherein --L2-R3 is (a); R3 is
Het' or Het2.
In an embodiment, the present invention relates to those compounds of Formula
(I) and
the pharmaceutically acceptable salts, and the solvates thereof, or any
subgroup thereof
as mentioned in any of the other embodiments, wherein --L2-R3 is (a); R3 is
Het'.
In an embodiment, the present invention relates to those compounds of Formula
(I) and
the pharmaceutically acceptable salts, and the solvates thereof, or any
subgroup thereof
as mentioned in any of the other embodiments, wherein --L2-R3 is (a); R3 is
Het'; and
Het' is azetidinyl optionally substituted as defined in any of the other
embodiments.
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In an embodiment, the present invention relates to those compounds of Formula
(I) and
the pharmaceutically acceptable salts, and the solvates thereof, or any
subgroup thereof
as mentioned in any of the other embodiments, wherein --L2-IV is (a); IV is
Het' or Het2;
.. Het' is a monocyclic heteroaryl selected from the group consisting of
pyridyl, 4-, 5- or
6-pyrimidinyl, pyrazinyl, pyridazinyl, pyrrolyl, pyrazolyl, and imidazolyl;
each of which
may be optionally substituted with one, two, or three substituents each
independently
selected from the group consisting of halo, -CN, -OR24, _NR251R25b5 and
Ci_zialkyl
optionally substituted with a substituent selected from the group consisting
of fluoro, -
.. CN, -OR265 _NR271R27b5 and ¨C(=0)NR271R27b; and
Het2 is a non-aromatic heterocyclyl selected from the group consisting of
azetidinyl,
pyrrolidinyl, and piperidinyl; each of which may be optionally substituted
with one, two,
or three substituents each independently selected from the group consisting of
halo, -CN,
_0R245 _NR251R25b5 and Ci_zialkyl optionally substituted with a substituent
selected from
the group consisting of fluoro, -CN, -0R265 _NR271R27b5 and ¨C(=0)NR271R27b.
In an embodiment, the present invention relates to those compounds of Formula
(I) and
the pharmaceutically acceptable salts, and the solvates thereof, or any
subgroup thereof
as mentioned in any of the other embodiments, wherein Ar is phenyl which may
be
optionally substituted with one, two, or three substituents each independently
selected
from the group consisting of halo, -
CN,
_0R245 _NR251R25b5 and Ci_zialkyl optionally substituted with a substituent
selected from
the group consisting of fluoro, -CN, -OR265 _NR271R27b5 and ¨C(=0)NR271R27b;
Het' is a monocyclic heteroaryl selected from the group consisting of pyridyl,
4-, 5- or
.. 6-pyrimidinyl, pyrazinyl, pyridazinyl, furanyl, thienyl, pyrrolyl,
pyrazolyl, imidazolyl,
4- or 5-thiazolyl, isothiazolyl, thiadiazolyl, and isoxazolyl; each of which
may be
optionally substituted with one, two, or three substituents each independently
selected
from the group consisting of halo, -CN, -OR245 _NR251R25b5 and Ci_zialkyl
optionally
substituted with a substituent selected from the group consisting of fluoro, -
CN, -0R26, -
NR271R27b5 and ¨C(=0)NR271R27b; and
Het2 is a monocyclic non-aromatic heterocyclyl optionally substituted with
one, two, or
three substituents each independently selected from the group consisting of
halo, -CN, -
0R245 _NR251R25b5 and Ci_zialkyl optionally substituted with a substituent
selected from
the group consisting of fluoro, -CN, -OR265 _NR271R27b5 and ¨C(=0)NR271R27b.
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In an embodiment, the present invention relates to those compounds of Formula
(I) and
the pharmaceutically acceptable salts, and the solvates thereof, or any
subgroup thereof
as mentioned in any of the other embodiments, wherein
--L2-R3 is (a), wherein
L2 is selected from the group consisting of >CR41R4b, and -CHR4aCHR5-; wherein
L2 is linked to a nitrogen atom of I);
R4a is selected from the group consisting of -C(=0)NR71R7b; and C-linked 4- to
7-
membered non-aromatic heterocyclyl containing at least one nitrogen, oxygen or
sulfur
atom;
.. R5 is selected from the group consisting of hydrogen; -0R6; -NR7aR7b;
-C(=0)NR71R7b; C1_4a1kyl optionally substituted with a substituent selected
from the
group consisting of fluoro, -CN, -0R8, and
-NR9aR9b;
R4b is selected from the group consisting of hydrogen and methyl; and
R3 is selected from the group consisting of Ar, Het', Het2, and a 7- to 10-
membered
saturated spirocarbobicyclic system.
In an embodiment, the present invention relates to those compounds of Formula
(I) and
the pharmaceutically acceptable salts, and the solvates thereof, or any
subgroup thereof
as mentioned in any of the other embodiments, wherein
--L2-R3 is (a), wherein
L2 is >CR41R4b; wherein
L2 is linked to a nitrogen atom of LI;
R4a is selected from the group consisting of -C(=0)NR71R7b; and C-linked 4- to
7-
membered non-aromatic heterocyclyl containing at least one nitrogen, oxygen or
sulfur
atom;
R4b is hydrogen; and
R3 is selected from the group consisting of Ar, Het', Het2, and a 7- to 10-
membered
saturated spirocarbobicyclic system.
In an embodiment, the present invention relates to those compounds of Formula
(I) and
the pharmaceutically acceptable salts, and the solvates thereof, or any
subgroup thereof
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as mentioned in any of the other embodiments, wherein
--L2-R3 is (a), wherein
L2 is >CR41R4b; wherein
L2 is linked to a nitrogen atom of I);
R4a is selected from the group consisting of -C(=0)NR71R7b; and C-linked 4- to
7-
membered non-aromatic heterocyclyl containing at least one nitrogen, oxygen or
sulfur
atom;
R4b is hydrogen; and
R3 is selected from the group consisting of Ar, and Het2.
In an embodiment, the present invention relates to those compounds of Formula
(I) and
the pharmaceutically acceptable salts, and the solvates thereof, or any
subgroup thereof
as mentioned in any of the other embodiments, wherein
--L2-R3 is (a), wherein
L2 is selected from the group consisting of >CR41R4b, and -CHR4aCHR5-; wherein
L2 is linked to a nitrogen atom of I);
R4a is -C(=0)NR71R7b;
R5 is selected from the group consisting of hydrogen; -0R6; -NR7aR7b;
-C(=0)NR71R7b; C1_4a1kyl optionally substituted with a substituent selected
from the
group consisting of fluoro, -CN, -0R8, and
-NR9aR9b;
R4b is selected from the group consisting of hydrogen and methyl; and
R3 is selected from the group consisting of Ar, Het', Het2, and a 7- to 10-
membered
saturated spirocarbobicyclic system.
In an embodiment, the present invention relates to those compounds of Formula
(I) and
the pharmaceutically acceptable salts, and the solvates thereof, or any
subgroup thereof
as mentioned in any of the other embodiments, wherein
--L2-R3 is (a), wherein
L2 is >CR4aR4b; wherein
L2 is linked to a nitrogen atom of LI;
R4a is -C(=0)NR71R7b;
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R4b is hydrogen; and
R3 is selected from the group consisting of Ar, Het', Het2, and a 7- to 10-
membered
saturated spirocarbobicyclic system.
In an embodiment, the present invention relates to those compounds of Formula
(I) and
the pharmaceutically acceptable salts, and the solvates thereof, or any
subgroup thereof
as mentioned in any of the other embodiments, wherein
--L2-R3 is (a), wherein
L2 is >CR41R4b; wherein
L2 is linked to a nitrogen atom of I);
R4a is -C(=0)NR71R7b;
R4b is hydrogen; and
R3 is selected from the group consisting of Ar, and Het2.
In an embodiment, the present invention relates to those compounds of Formula
(I) and
the pharmaceutically acceptable salts, and the solvates thereof, or any
subgroup thereof
as mentioned in any of the other embodiments, wherein
--L2-R3 is (a), wherein
L2 is selected from the group consisting of >CR41R4b, and -CHR4aCHR5-; wherein
L2 is linked to a nitrogen atom of I);
R4a is C-linked 4- to 7-membered non-aromatic heterocyclyl containing at least
one
nitrogen, oxygen or sulfur atom;
R5 is selected from the group consisting of hydrogen; -0R6; -NR71R7b;
-C(=0)NR71R7b; C1_4a1kyl optionally substituted with a substituent selected
from the
group consisting of fluoro, -CN, -0R8, and
-NR9aR9b;
R4b is selected from the group consisting of hydrogen and methyl; and
R3 is selected from the group consisting of Ar, Het', Het2, and a 7- to 10-
membered
saturated spirocarbobicyclic system.
In an embodiment, the present invention relates to those compounds of Formula
(I) and
the pharmaceutically acceptable salts, and the solvates thereof, or any
subgroup thereof
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as mentioned in any of the other embodiments, wherein
--L2-R3 is (a), wherein
L2 is >CR41R4b; wherein
L2 is linked to a nitrogen atom of I);
R4a is C-linked 4- to 7-membered non-aromatic heterocyclyl containing at least
one
nitrogen, oxygen or sulfur atom;
R4b is hydrogen; and
R3 is selected from the group consisting of Ar, Het', Het2, and a 7- to 10-
membered
saturated spirocarbobicyclic system.
In an embodiment, the present invention relates to those compounds of Formula
(I) and
the pharmaceutically acceptable salts, and the solvates thereof, or any
subgroup thereof
as mentioned in any of the other embodiments, wherein
--L2-R3 is (a), wherein
L2 is >CR41R4b; wherein
L2 is linked to a nitrogen atom of I);
R4a is C-linked 4- to 7-membered non-aromatic heterocyclyl containing at least
one
nitrogen, oxygen or sulfur atom;
R4b is hydrogen; and
R3 is selected from the group consisting of Ar, and Het2.
In an embodiment, the present invention relates to those compounds of Formula
(I) and
the pharmaceutically acceptable salts, and the solvates thereof, or any
subgroup thereof
as mentioned in any of the other embodiments, wherein L2 is linked to a
nitrogen atom
ofLl.
In an embodiment, the present invention relates to those compounds of Formula
(I) and
the pharmaceutically acceptable salts, and the solvates thereof, or any
subgroup thereof
as mentioned in any of the other embodiments, wherein
--L2-R3 is selected from the group consisting of
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¨0
'0.........0 Z )......, )......"0
N N
I ,
,
, N
I
F
1 F __
18 Ri8 R18
R , ,
F
--4
N , N
I ,
, I ,
R18 ,and R18
5.
In an embodiment, the present invention relates to those compounds of Formula
(I) and
the pharmaceutically acceptable salts, and the solvates thereof, or any
subgroup thereof
as mentioned in any of the other embodiments, wherein
--L2-R3 is selected from the group consisting of
¨o
).......eo ).......,
N N
N
118
RI1 I
' 8 R 18
R 5 5
F F
F
N N
1
RI
R 18 ,and 18
5=
wherein V is hydrogen or methyl.
In an embodiment, the present invention relates to those compounds of Formula
(I) and
the pharmaceutically acceptable salts, and the solvates thereof, or any
subgroup thereof
as mentioned in any of the other embodiments, wherein
L' represents a N-linked 7- to 10-membered saturated spiroheterobicyclic
system
containing one or two N-atoms selected from the group consisting of (a), (b),
(c), (d), (e),
(f), (g), (h), and (i)
O
i
0N ciNil yiN" -
N N N N
i a (a) i a (b) a , (c) a i (d)
i 1
Q i N
g )
--... .---
N N N
a i (e) a i (0 a i (g) a
1 (h)
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a' (i)
wherein a represents the position of linkage to the thienopyrimidinyl
heterocycle.
In an embodiment, the present invention relates to those compounds of Formula
(I) and
the pharmaceutically acceptable salts, and the solvates thereof, or any
subgroup thereof
as mentioned in any of the other embodiments, wherein Het2 is monocyclic
heterocyclyl
optionally substituted with one, two, or three substituents as described in
the other
embodiments.
In an embodiment, the present invention relates to those compounds of Formula
(I) and
the pharmaceutically acceptable salts, and the solvates thereof, or any
subgroup thereof
as mentioned in any of the other embodiments, wherein Het2 is a non-aromatic
heterocyclyl selected from azetidinyl, oxetanyl, tetrahydrofuranyl,
piperidinyl,
coN
N H
and
tetrahydropyranyl, tetrahydro-2H-thiopyranyl 1,1-dioxide,
each of which are optionally substituted with one, two, or three substituents
as described
in the other embodiments.
In an embodiment, the present invention relates to those compounds of Formula
(I) and
the pharmaceutically acceptable salts, and the solvates thereof, or any
subgroup thereof
as mentioned in any of the other embodiments, wherein
Het2 is a non-aromatic heterocyclyl selected from
0 N /0 \ /N \
0
0
NH
each of which are optionally substituted with one, two, or three substituents
as described
in the other embodiments.
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Particular compounds of Formula (I) are compounds 82, 84, 273, and 274,
including
the stereoisomeric forms, the pharmaceutically acceptable salts thereof, in
particular the
hydrochloride salts thereof, and the solvates thereof.
Particular compounds of Formula (I) are compounds 82, 84, 273, and 274.
In an embodiment the compound of Formula (I) is selected from the group
consisting
of any of the exemplified compounds,
and the free bases, the pharmaceutically acceptable addition salts, and the
solvates
thereof.
All possible combinations of the above-indicated embodiments are considered to
be
.. embraced within the scope of this invention.
METHODS FOR THE PREPARATION OF COMPOUNDS OF FORMULA (I)
In this section, as in all other sections unless the context indicates
otherwise, references
to Formula (I) also include all other sub-groups and examples thereof as
defined herein.
The general preparation of some typical examples of the compounds of Formula
(I) is
described hereunder and in the specific examples, and are generally prepared
from
starting materials which are either commercially available or prepared by
standard
synthetic processes commonly used by those skilled in the art. The following
schemes
are only meant to represent examples of the invention and are in no way meant
to be a
limit of the invention.
Alternatively, compounds of the present invention may also be prepared by
analogous
reaction protocols as described in the general schemes below, combined with
standard
synthetic processes commonly used by those skilled in the art of organic
chemistry.
The skilled person will realize that in the reactions described in the
Schemes, although
this is not always explicitly shown, it may be necessary to protect reactive
functional
groups (for example hydroxy, amino, or carboxy groups) where these are desired
in the
final product, to avoid their unwanted participation in the reactions. For
example in
Scheme 1, the NH moiety on the Ll N-linked 7- to 10-membered saturated
spiroheterobicyclic system containing one or two N-atoms can be protected with
a tert-
butoxycarbonyl protecting group. In general, conventional protecting groups
can be used
in accordance with standard practice. The protecting groups may be removed at
a
convenient subsequent stage using methods known from the art. This is
illustrated in the
specific examples.
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The skilled person will realize that in the reactions described in the
Schemes, it may be
advisable or necessary to perform the reaction under an inert atmosphere, such
as for
example under N2-gas atmosphere.
It will be apparent for the skilled person that it may be necessary to cool
the reaction
mixture before reaction work-up (refers to the series of manipulations
required to isolate
and purify the product(s) of a chemical reaction such as for example
quenching, column
chromatography, extraction).
The skilled person will realize that heating the reaction mixture under
stirring may
enhance the reaction outcome. In some reactions microwave heating may be used
instead
of conventional heating to shorten the overall reaction time.
The skilled person will realize that another sequence of the chemical
reactions shown in
the Schemes below, may also result in the desired compound of Formula (I).
The skilled person will realize that intermediates and final compounds shown
in the
Schemes below may be further functionalized according to methods well-known by
the
person skilled in the art. The intermediates and compounds described herein
can be
isolated in free form or as a salt.
SCHEME 1
In general, compounds of Formula (I) wherein all variables are defined
according to the
scope of the present invention, can be prepared according to the following
reaction
- - NN - -
Scheme 1. In Scheme 1, represents L1 as a 7- to 10-membered saturated
spiroheterobicyclic system containing two N-atoms and which is N-linked to the
thienopyrimidinyl heterocycle, LG1 and LG2 each represent a suitable leaving
group,
such as for example halo or methanesulfonyl; PG' represents a suitable
protecting group,
such as for example tert-butyloxycarbonyl; R3a¨PG2 represents an R3 as defined
in
Formula (I) with an appropriate protecting group, such as for example tert-
butyloxycarbonyl, when the R3 substituent bears an amino group. The X in
formula (XI)
represents CH or N (in formula (XI) L2 can be linked to a carbon or a N-atom).
All other
variables in Scheme 1 are defined according to the scope of the present
invention.
In Scheme 1, the following reaction conditions apply:
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R1 R1
H-001-PG1
LG _____________________________________________________________________ S
00-PG1
I
N
yN yN (II) (IV)
R2 R2
RI
H_Nx-L2-R3
8
L 2 r< 2
LG2., 2,R3..! 2
(XI) 2
NN L PG
(VII)
R2 4
Ri
Ri
L 2 R
3 LG2 R3
D S 00-
H
(V
NyN (I) 3 NyN
(v)
R2
R2 0
7 Ri ell"R3 (X)
S yN N=CH-R3
NN (v)
R2
1: at a suitable temperature such as ranged from rt to 90 C, in the presence
of a suitable
base such as for example diisopropylethylamine, in a suitable solvent such as
for example
acetonitrile or isopropanol or ethanol;
2: at a suitable temperature range such as for example from 0 C to room
temperature, in
the presence of suitable cleavage conditions, such as for example an acid such
as HC1 or
trifluoroacetic acid in a suitable solvent such as acetonitrile or
dichloromethane when
PG' is tert-butyloxycarbonyl;
Alternatively, at a suitable temperature such as for example room temperature
in a
suitable solvent such as acetic acid
3: at a suitable temperature such as for example room temperature or reflux,
in the
presence of a suitable base such as for example potassium carbonate or 1,8-
Diazabicyclo[5.4.0]undec-7-ene, in a suitable solvent such as for example
acetonitrile or
DMSO;
4: at a suitable temperature such as for example room temperature or 90 C, in
the
presence of a suitable base such as for example potassium carbonate or 1,8-
Diazabicyclo[5.4.0]undec-7-ene, in a suitable solvent such as for example
acetonitrile or
DMSO;
5: at a suitable reaction temperature range such as for example from 0 C to
room
temperature, in the presence of suitable cleavage conditions, such as for
example an acid
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such as HC1 or trifluoroactic acid in a suitable solvent such as acetonitrile
or
dichloromethane when PG2 is tert-butyloxycarbonyl.
6: at a suitable temperature such as for example at room temperature,
eventually in the
presence of a suitable base such as for example trimethylamine or a suitable
acid such as
for example acetic acid, in a suitable solvent such as for example anhydrous
dichloromethane, dichloroethane or tetrahydropyrane;
7: at a suitable temperature, for example room temperature, in the presence of
a suitable
reducing agent, such as for example NaBH(OAc)3, in a suitable solvent such as
dichloromethane, dichloroethane or tetrahydropyran; yielding a compound of
Formula
(I) wherein L1 is a N-linked 7- to 10-membered saturated spiroheterobicyclic
system
containing two N-atoms and L2 is CH2.
Steps 6 and 7 can conveniently be performed as a one-pot procedure.
Alternativelly, step 6 and 7 can be performed in the presence of a suitable
acid such as
for example acetic acid, a suitable catalyst such as platinium oxide, in a
suitable solvent
such as for example ethanol at a suitable temperature such as for exemple 60
C;
8: at a suitable temperature such as for example at 90 C, in the presence of
a suitable
base such as for example diisopropylethylamine, in a suitable solvent such as
for example
acetonitrile or isopropanol. In step 8, reagent of Formula (XI), X represents
CH or N,
and L2 and R3 are as defined according to the scope of the invention. Reagents
of
Formula (XI) are either commercially available or can be prepared by methods
known to
the skilled person from commercially available starting materials, e.g. by
appropriate
protection/deprotection steps and functional group interconversion, from
starting
materials, such as 2-azaspiro[3.3]heptan-6-ol (CAS[1256352-97-2]).
SCHEME 2
Intermediates of Formula (II), wherein R2 is methyl, can be prepared according
to the
following reaction Scheme 2, wherein LG1 represents a suitable leaving group,
such as
for example halo or methanesulfonyl. All other variables in Scheme 2 are
defined
according to the scope of the present invention.
In Scheme 2, the following reaction conditions apply:
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R R R Ri
So
1
-3im=
1 2 SOH SLG
ONH N H2 (X) ONH N H2 (XI) Ny N (xii) N N
y (mu)
NH2 cH3 cH3 cH3
1: at a suitable temperature such as for example at reflux temperature, in the
presence of
acetic anhydride and a suitable base such as for example trimethylamine, in a
suitable
solvent such as for example toluene;
2: at a suitable temperature such as for example at reflux temperature, in the
presence of
a suitable base such as potassium hydroxide, in a suitable solvent such as for
example
ethanol;
3: under suitable reaction conditions to form a leaving group, such as for
example, chloro,
for example by reaction with phosphoryl trichloride at a suitable temperature
such as 110
C.
SCHEME 3
In general, compounds of Formula (I-a) wherein the variables are defined
according to
the scope of the present invention, but wherein L2 is limited to L2a (the
options that can
be obtained by this Scheme), can be prepared according to the following
reaction Scheme
3. All other variables in Scheme 3 are defined according to the scope of the
present
invention or as defined before.
In Scheme 3, the following reaction conditions apply:
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R1
.?........r.,N N-H
I
N.õ.....e.
I 2 (V)
R
0
0
R4a)y3
a....õ.1, I
R4 R
1 1 R
(AV)
(XV)
R1
R1
R4a R4a
..,...,(N N
S .............1.......,N N
I R3 I R
S¨ 3
N,,,,,,,,õ N N,....e.õ N
(XVI)
I 2
I 2 R
R
\ /
R1
I 1
.,..-- " 2 ....-R3
L a
I
N..,,,,,,,õ N (I-a)
I 2
R
1: at a suitable temperature such as for example room temperature or 45 C, in
the
presence of titanium (IV) ethoxide or titanium (IV) isopropoxide, in a
suitable solvent
such as for example tetrahydropyrane, dichloroethane or a mixture of
dichloroethane and
methanol;
Alternatively, at a suitable temperature such as for example room temperature,
with or
without a suitable acid such as for example trifluoroacetic acid, in a
suitable solvent such
as for example tetrahydropyrane;
2: at a suitable temperature such as for example room temperature, in the
presence of a
suitable reducting agent such as for example sodium borohydride, sodium
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triacetoxyborohydride or sodium cyanoborohydride, in a suitable solvent such
as for
example tetrahydropyrane, dichloroethane or a mixture of dichloroethane and
methanol;
Steps 1 and 2 can be performed as a one-pot procedure.
SCHEME 4
In general, compounds of Formula (I-b) wherein R4a. is rectricted to R4a1
being C1_4alkyl or a C-
linked 4- to 7-membered non-aromatic heterocyclyl containing at least one
nitrogen,
oxygen or sulfur atom can be prepared according to the following reaction
Scheme 4. In
Scheme 4, halo means chloro, bromo or iodo. All other variables in Scheme 4
are defined
according to the scope of the present invention or as defined before.
In Scheme 4, the following reaction conditions apply:
R1
R
Sfl
0
HR3
SyN
CN-H
N=\ 3
__________________________________________ 31. IN
N
I 2
I 2
(V) (W111)
R1
r,4a1 ,
¨LI
R4a1
_____________________________ Syr(MON_(
Halo R3
4a1 N
R ¨Mg
I 2
(I-b)
00()
1: at a suitable temperature such as for example room temperature or 45 C, in
the
presence of titanium (IV) ethoxide or titanium (IV) isopropoxide, in a
suitable solvent
such as for example tetrahydropyran;
2: at a suitable temperature ranged from 0 C to room temperature, in a
suitable solvent
such as for example tetrahydrofurane.
Steps 1 and 2 can be performed as a one-pot procedure.
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SCHEME 5
In general, compounds of Formula (Ic) wherein R3 is rectricted to R3' being
R18a
118
can be prepared according to the following reaction Scheme 5. All other
variables in
Scheme 5 are defined according to the scope of the present invention or as
defined before.
In Scheme 5, L2 is linked to a N-atom of LI.
In Scheme 5, the following reaction conditions apply:
R18a
R1
R1
118 0 H
S$yN N-H (XXI) S
LLL2R3c
1 NN
NN (IC)
12
12
(V)
1: at a suitable temperature, such as for example room temperature, in the
presence of a
suitable acid coupling agent, such as for example
14bis(dimethylamino)methylene]-
1H-benzotriazoliumhexafluorophosphate(1-)3-oxide (HBTU) or 1-
[Bis(dimethylamino)methylene]-1H-1,2,3-triazolo [4,5-b]pyridinium 3-oxide
hexafluorophosphate (HATU), in the presence of a suitable base such as for
example
N-ethyl-N-(1-methylethyl)-2-propanamine (DIPEA), in a suitable solvent such as
N,N-
dimethylformamide (DMF);
SCHEME 6
In general, compounds of Formula (Id) wherein L2 is rectricted to SO2, can be
prepared
according to the following reaction Scheme 6. All other variables in Scheme 6
are defined
according to the scope of the present invention or as defined before. In
Scheme 6, L2
(>S02 in Scheme 6) is linked to a N-atom of LI.
In Scheme 6, the following reaction conditions apply:
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Ri Ri
S ----- N
/
I
N-H 0
II
CI¨R3
II
0 OM
¨a L 1 0
...../ .., //
S
il i 3
NN 0 R
N.......õ... 1
I 2 I 2
R
R
N)
(Id)
1: at a suitable temperature, for example room temperature, in the presence of
a suitable
base such as for example potassium carbonate, in a suitable solvent such as
for example
acetonitrile.
SCHEME 7
In general, compounds of Formula (le) and (If) can be prepared according to
the
following reaction Scheme 7.Both in (le) and (If) the L2 part of the molecule
is linked to
a nitrogen atom of LI. All other variables are defined according to the scope
of the present
invention or as defined before.
In Scheme 7, the following reaction conditions apply:
R1
R4a >R
R1 Ri
R4b
I
-.........r R4b
3
SyN N-H
$ Li
...............,R
(0(111) $ and/or s / ¨a S Ll
I
OH
I 2 N,.......
R I 2
R
I 2
(V) R
(le) (If)
1: at a suitable temperature such as for example 60 C, in a suitable solvent
such as for example
ethanol.
It will be appreciated that where appropriate functional groups exist,
compounds of various
formulae or any intermediates used in their preparation may be further
derivatised by one or more
standard synthetic methods employing condensation, substitution, oxidation,
reduction, or
cleavage reactions. Particular substitution approaches include conventional
alkylation, arylation,
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heteroarylation, acylation, sulfonylation, halogenation, nitration,
formylation and coupling
procedures.
The compounds of Formula (I) may be synthesized in the form of racemic
mixtures of
enantiomers which can be separated from one another following art-known
resolution
procedures. The racemic compounds of Formula (I) containing a basic nitrogen
atom
may be converted into the corresponding diastereomeric salt forms by reaction
with a
suitable chiral acid. Said diastereomeric salt forms are subsequently
separated, for
example, by selective or fractional crystallization and the enantiomers are
liberated
therefrom by alkali. An alternative manner of separating the enantiomeric
forms of the
compounds of Formula (I) involves liquid chromatography using a chiral
stationary
phase. Said pure stereochemically isomeric forms may also be derived from the
corresponding pure stereochemically isomeric forms of the appropriate starting
materials, provided that the reaction occurs stereospecifically.
In the preparation of compounds of the present invention, protection of remote
functionality (e.g., primary or secondary amine) of intermediates may be
necessary. The
need for such protection will vary depending on the nature of the remote
functionality
and the conditions of the preparation methods. Suitable amino-protecting
groups (NH-
Pg) include acetyl, trifluoroacetyl, t-butoxycarbonyl (Boc), benzyloxycarbonyl
(CBz)
and 9-fluorenylmethyleneoxycarbonyl (Fmoc). The need for such protection is
readily
determined by one skilled in the art. For a general description of protecting
groups and
their use, see T. W. Greene and P. G. M. Wuts, Protective Groups in Organic
Synthesis,
4th ed., Wiley, Hoboken, New Jersey, 2007.
PHARMACOLOGY
It has been found that the compounds of the present invention block the
interaction of
menin with MLL proteins and oncogenic MLL fusion proteins. Therefore the
compounds
according to the present invention and the pharmaceutical compositions
comprising such
compounds may be useful for the treatment or prevention, in particular
treatment, of
diseases such as cancer, myelodysplastic syndrome (MDS) and diabetes.
In particular, the compounds according to the present invention and the
pharmaceutical
compositions thereof may be useful in the treatment or prevention of cancer.
According
to one embodiment, cancers that may benefit from a treatment with menin/MLL
inhibitors of the invention comprise leukemias, myeloma or a solid tumor
cancer (e.g.
prostate cancer, lung cancer, breast cancer, pancreatic cancer, colon cancer,
liver cancer,
melanoma and glioblastoma, etc.). In some embodiments, the leukemias include
acute
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leukemias, chronic leukemias, myeloid leukemias, myelogeneous leukemias,
lymphoblastic leukemias, lymphocytic leukemias, Acute myelogeneous leukemias
(AML), Chronic myelogenous leukemias (CML), Acute lymphoblastic leukemias
(ALL), Chronic lymphocytic leukemias (CLL), T cell prolymphocytic leukemias (T-
PLL), Large granular lymphocytic leukemia, Hairy cell leukemia (HCL), MLL-
rearranged leukemias, MLL-PTD leukemias, MLL amplified leukemias, MLL-positive
leukemias, leukemias exphibiting HOXIMEIS1 gene expression signatures etc.
Hence, the invention relates to compounds of Formula (I), the tautomers and
the
stereoisomeric forms thereof, and the pharmaceutically acceptable salts, and
the solvates
thereof, for use as a medicament.
The invention also relates to the use of a compound of Formula (I), a tautomer
or a
stereoisomeric form thereof, or a pharmaceutically acceptable salt, or a
solvate thereof,
or a pharmaceutical composition according to the invention, for the
manufacture of a
medicament.
The present invention also relates to a compound of Formula (I), a tautomer or
a
stereoisomeric form thereof, or a pharmaceutically acceptable salt, or a
solvate thereof,
or a pharmaceutical composition according to the invention, for use in the
treatment,
prevention, amelioration, control or reduction of the risk of disorders
associated with the
interaction of menin with MLL proteins and oncogenic MLL fusion proteins in a
mammal, including a human, the treatment or prevention of which is affected or
facilitated by blocking the interaction of menin with MLL proteins and
oncogenic MLL
fusion proteins.
Also, the present invention relates to the use of a compound of Formula (I), a
tautomer
or a stereoisomeric form thereof, or a pharmaceutically acceptable salt, or a
solvate
thereof, or a pharmaceutical composition according to the invention, for the
manufacture
of a medicament for treating, preventing, ameliorating, controlling or
reducing the risk
of disorders associated with the interaction of menin with MLL proteins and
oncogenic
MLL fusion proteins in a mammal, including a human, the treatment or
prevention of
which is affected or facilitated by blocking the interaction of menin with MLL
proteins
and oncogenic MLL fusion proteins.
The invention also relates to a compound of Formula (I), a tautomer or a
stereoisomeric
form thereof, or a pharmaceutically acceptable salt, or a solvate thereof, for
use in the
treatment or prevention of any one of the diseases mentioned hereinbefore.
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The invention also relates to a compound of Formula (I), a tautomer or a
stereoisomeric
form thereof, or a pharmaceutically acceptable salt, or a solvate thereof, for
use in treating
or preventing any one of the diseases mentioned hereinbefore.
The invention also relates to the use of a compound of Formula (I), a tautomer
or a
.. stereoisomeric form thereof, or a pharmaceutically acceptable salt, or a
solvate thereof,
for the manufacture of a medicament for the treatment or prevention of any one
of the
disease conditions mentioned hereinbefore.
The compounds of the present invention can be administered to mammals,
preferably
humans, for the treatment or prevention of any one of the diseases mentioned
.. hereinbefore.
In view of the utility of the compounds of Formula (I), the tautomers and the
stereoisomeric forms thereof, and the pharmaceutically acceptable salts, and
the solvates
thereof, there is provided a method of treating warm-blooded animals,
including humans,
suffering from any one of the diseases mentioned hereinbefore.
.. Said method comprises the administration, i.e. the systemic or topical
administration,
preferably oral administration, of a therapeutically effective amount of a
compound of
Formula (I), a tautomer or a stereoisomeric form thereof, or a
pharmaceutically
acceptable salt, or a solvate thereof, to warm-blooded animals, including
humans.
Therefore, the invention also relates to a method for the treatment or
prevention of any
.. one of the diseases mentioned hereinbefore comprising administering a
therapeutically
effective amount of compound according to the invention to a patient in need
thereof
One skilled in the art will recognize that a therapeutically effective amount
of the
compounds of the present invention is the amount sufficient to have
therapeutic activity
and that this amount varies inter alias, depending on the type of disease, the
concentration of the compound in the therapeutic formulation, and the
condition of the
patient. Generally, the amount of a compound of the present invention to be
administered as a therapeutic agent for treating the disorders referred to
herein will be
determined on a case by case by an attending physician.
Those of skill in the treatment of such diseases could determine the effective
therapeutic
daily amount from the test results presented hereinafter. An effective
therapeutic daily
amount would be from about 0.005 mg/kg to 100 mg/kg, in particular 0.005 mg/kg
to 50
mg/kg, in particular 0.01 mg/kg to 50 mg/kg body weight, more in particular
from 0.01
mg/kg to 25 mg/kg body weight, preferably from about 0.01 mg/kg to about 15
mg/kg,
more preferably from about 0.01 mg/kg to about 10 mg/kg, even more preferably
from
about 0.01 mg/kg to about 1 mg/kg, most preferably from about 0.05 mg/kg to
about 1
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mg/kg body weight. A particular effective therapeutic daily amount might be 1
mg/kg
body weight, 2 mg/kg body weight, 4 mg/kg body weigth, or 8 mg/kg body weight.
The
amount of a compound according to the present invention, also referred to
herein as the
active ingredient, which is required to achieve a therapeutically effect may
vary on case-
by-case basis, for example with the particular compound, the route of
administration, the
age and condition of the recipient, and the particular disorder or disease
being treated. A
method of treatment may also include administering the active ingredient on a
regimen
of between one and four intakes per day. In these methods of treatment the
compounds
according to the invention are preferably formulated prior to administration.
As
described herein below, suitable pharmaceutical formulations are prepared by
known
procedures using well known and readily available ingredients.
The present invention also provides compositions for preventing or treating
the disorders
referred to herein. Said compositions comprising a therapeutically effective
amount of a
compound of Formula (I), a tautomer or a stereoisomeric form thereof, or a
pharmaceutically acceptable salt, or a solvate thereof, and a pharmaceutically
acceptable
carrier or diluent.
While it is possible for the active ingredient to be administered alone, it is
preferable to
present it as a pharmaceutical composition. Accordingly, the present invention
further
provides a pharmaceutical composition comprising a compound according to the
present
invention, together with a pharmaceutically acceptable carrier or diluent. The
carrier or
diluent must be "acceptable" in the sense of being compatible with the other
ingredients
of the composition and not deleterious to the recipients thereof
The pharmaceutical compositions of this invention may be prepared by any
methods well
known in the art of pharmacy, for example, using methods such as those
described in
Gennaro et al. Remington's Pharmaceutical Sciences (18th ed., Mack Publishing
Company, 1990, see especially Part 8 : Pharmaceutical preparations and their
Manufacture). A therapeutically effective amount of the particular compound,
in base
form or salt form, as the active ingredient is combined in intimate admixture
with a
pharmaceutically acceptable carrier, which may take a wide variety of forms
depending
on the form of preparation desired for administration. These pharmaceutical
compositions are desirably in unitary dosage form suitable, preferably, for
systemic
administration such as oral, percutaneous or parenteral administration; or
topical
administration such as via inhalation, a nose spray, eye drops or via a cream,
gel,
shampoo or the like. For example, in preparing the compositions in oral dosage
form, any
of the usual pharmaceutical media may be employed, such as, for example,
water,
glycols, oils, alcohols and the like in the case of oral liquid preparations
such as
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suspensions, syrups, elixirs and solutions: or solid carriers such as
starches, sugars,
kaolin, lubricants, binders, disintegrating agents and the like in the case of
powders, pills,
capsules and tablets. Because of their ease in administration, tablets and
capsules
represent the most advantageous oral dosage unit form, in which case solid
pharmaceutical carriers are obviously employed. For parenteral compositions,
the carrier
will usually comprise sterile water, at least in large part, though other
ingredients, for
example, to aid solubility, may be included. Injectable solutions, for
example, may be
prepared in which the carrier comprises saline solution, glucose solution or a
mixture of
saline and glucose solution. Injectable suspensions may also be prepared in
which case
appropriate liquid carriers, suspending agents and the like may be employed.
In the
compositions suitable for percutaneous administration, the carrier optionally
comprises
a penetration enhancing agent and/or a suitable wettable agent, optionally
combined with
suitable additives of any nature in minor proportions, which additives do not
cause any
significant deleterious effects on the skin. Said additives may facilitate the
administration
to the skin and/or may be helpful for preparing the desired compositions.
These
compositions may be administered in various ways, e.g., as a transdermal
patch, as a spot-
on or as an ointment.
It is especially advantageous to formulate the aforementioned pharmaceutical
compositions in dosage unit form for ease of administration and uniformity of
dosage.
Dosage unit form as used in the specification and claims herein refers to
physically
discrete units suitable as unitary dosages, each unit containing a
predetermined quantity
of active ingredient calculated to produce the desired therapeutic effect in
association
with the required pharmaceutical carrier. Examples of such dosage unit forms
are tablets
(including scored or coated tablets), capsules, pills, powder packets, wafers,
injectable
solutions or suspensions, teaspoonfuls, tablespoonfuls and the like, and
segregated
multiples thereof.
The present compounds can be used for systemic administration such as oral,
percutaneous or parenteral administration; or topical administration such as
via
inhalation, a nose spray, eye drops or via a cream, gel, shampoo or the like.
The
compounds are preferably orally administered. The exact dosage and frequency
of
administration depends on the particular compound of Formula (I) used, the
particular
condition being treated, the severity of the condition being treated, the age,
weight, sex,
extent of disorder and general physical condition of the particular patient as
well as other
medication the individual may be taking, as is well known to those skilled in
the art.
Furthermore, it is evident that said effective daily amount may be lowered or
increased
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depending on the response of the treated subject and/or depending on the
evaluation of
the physician prescribing the compounds of the instant invention.
The compounds of the present invention may be administered alone or in
combination
with one or more additional therapeutic agents. Combination therapy includes
administration of a single pharmaceutical dosage formulation which contains a
compound according to the present invention and one or more additional
therapeutic
agents, as well as administration of the compound according to the present
invention and
each additional therapeutic agent in its own separate pharmaceutical dosage
formulation.
For example, a compound according to the present invention and a therapeutic
agent may
be administered to the patient together in a single oral dosage composition
such as a
tablet or capsule, or each agent may be administered in separate oral dosage
formulations.
Therefore, an embodiment of the present invention relates to a product
containing as first
active ingredient a compound according to the invention and as further active
ingredient
one or more anticancer agent, as a combined preparation for simultaneous,
separate or
sequential use in the treatment of patients suffering from cancer.
The one or more other medicinal agents and the compound according to the
present
invention may be administered simultaneously (e.g. in separate or unitary
compositions)
or sequentially in either order. In the latter case, the two or more compounds
will be
administered within a period and in an amount and manner that is sufficient to
ensure
that an advantageous or synergistic effect is achieved. It will be appreciated
that the
preferred method and order of administration and the respective dosage amounts
and
regimes for each component of the combination will depend on the particular
other
medicinal agent and compound of the present invention being administered,
their route
of administration, the particular condition, in particular tumour being
treated and the
particular host being treated. The optimum method and order of administration
and the
dosage amounts and regime can be readily determined by those skilled in the
art using
conventional methods and in view of the information set out herein.
The weight ratio of the compound according to the present invention and the
one or more
other anticancer agent(s) when given as a combination may be determined by the
person
skilled in the art. Said ratio and the exact dosage and frequency of
administration
depends on the particular compound according to the invention and the other
anticancer
agent(s) used, the particular condition being treated, the severity of the
condition being
treated, the age, weight, gender, diet, time of administration and general
physical
condition of the particular patient, the mode of administration as well as
other medication
the individual may be taking, as is well known to those skilled in the art.
Furthermore,
it is evident that the effective daily amount may be lowered or increased
depending on
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the response of the treated subject and/or depending on the evaluation of the
physician
prescribing the compounds of the instant invention. A particular weight ratio
for the
present compound of Formula (I) and another anticancer agent may range from
1/10 to
10/1, more in particular from 1/5 to 5/1, even more in particular from 1/3 to
3/1.
The following examples further illustrate the present invention.
EXAMPLES
Several methods for preparing the compounds of this invention are illustrated
in the
following examples. Unless otherwise noted, all starting materials were
obtained from
commercial suppliers and used without further purification.
Hereinafter, the terms : 'ACN or `MeCN' means acetonitrile, `DCM' means
dichloromethane, `DIPEA' means N,N-diisopropylethylamine, `DIPE or `DiPE"
means
diisopropyl ether, 'h' means hours(s), 'min' means minute(s), 'DMF' means
dimethylformamide, 'DSC' means differential scanning calorimetry, 'TEA or
'Et3N'
means triethyl amine, 'Et20' means diethyl ether, 'Et0Ac' or 'EA' means ethyl
acetate,
'Et0H' means ethanol, 'FIPLC' means High-performance Liquid Chromatography,
'iPrOH' means isopropyl alcohol, IC/MS' means Liquid Chromatography/Mass
Spectrometry, `MeOH' means methanol, `NMR' means Nuclear Magnetic Resonance,
or `RT' means room temperature, `SFC' means supercritical fluid
chromatography,
'OR' means optical rotation, 'sat. aq.' means saturated aqueous. 'AcC1' means
acetyl
chloride, 'AcOH' or 'HOAc' means acetic acid, 'BOC' or 'Boc' means tert-
butyloxycarbonyl, 'Celite means diatomaceous earth, 'CH3COONH4' means
ammonium acetate, 'COMUCY means (1-
Cyano-2-ethoxy-2-
oxoethylidenaminooxy)dimethylamino-morpho lino -carb enium
hexafluorophosphate,
'CO2' means carbon dioxide, `DCE' means dichloroethane, 'DMAP' means
dimethylaminopyridine, 'DMS0' means dimethyl sulfoxyde, `DBU' means 1,8-
diazabicyc lo [5.4.0]undec ene-7, ED
CLHC l' means 1-(3-Dimethylaminopropy1)-3-
ethylcarbodiimide hydrochloride, `ee' means enantiomeric excess, 'eq.' or
'equiv.'
means equivalent(s), 'EtMgBr' means ethyl magnesium bromide, 'Et20' means
diethyl
ether, 'Et0Ac' means ethyl acetate, 'Et3N' or 'TEA' means triethylamine,
'Et0H' means
ethanol, 'h' means hours(s), 'HATU' means 0-(7-Azabenzotriazol-1-y1)-N,N,N,N-
tetramethyluronium hexafluorophosphate, 'FIC1' means hydrochloric acid, 'HOBT'
means N-Hydroxybenzotrizole monohydrate, 'H20' means water, 'iPrMgC1' means
isopropyl magnesium chloride, 'iPrNH2' means isopropylamine, `1(2CO3' means
potassium carbonate, `Me-THF' means 2-methyl-tetrahydrofuran, `MeMgBe or
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`CH3MgBe means methyl magnesium bromide, `MeOH' means methanol, 'MgSO4'
means magnesium sulfate, 'min' means minute(s), NaBH(OAc)3' means sodium
triacetoxyborohydride, NaBH3CN' means sodium cyanoborohydride, Na2CO3' means
sodium carbonate, `NaH' means sodium hydride, NaHCO3' means sodium
hydrogenocarloonate, 'NaOH' means potassium hydroxide, 'INTa2SO4' means sodium
sulfate, '1\TH4C1' means ammonium chloride, '1\1144HCO3' means ammonium
bicarbonate, NH4OH' means ammonia solution 30% aqueous, 'Quant. or quant'
means
quantitative, at' means retention time, `SFC' means supercritical fluid
chromatography,
'T' means temperature, 'TBAF' means tetrabutylammonium fluoride, 'TBDMS' or
`SMDBT' means tert-butyldimethylsilyl, 'TFA' or `CF3COOH' means
trifluoroacetic
acid, `THF' means tetrahydrofuran, 'Ti(OEt)4' means titanium ethoxyde,
'Ti(OiPr)4'
means titanium isopropoxide, 'v.' means volume, 'F3C' or `CF3' means
trifluoromethyl,
'FIBTU' means 1-
[bis(dimethylamino)methylene]-1H-
benzotriazo liumhexafluoropho sphate(1 -)3 -oxide.
As understood by a person skilled in the art, compounds synthesised using the
protocols
as indicated may exist as a solvate e.g. hydrate, and/or contain residual
solvent or minor
impurities. Compounds isolated as a salt form, may be integer stoichiometric
i.e. mono-
or di-salts, or of intermediate stoichiometry.
The stereochemical configuration for centres in some compounds may be
designated
"R" or "S" when the mixture(s) was separated; for some compounds, the
stereochemical configuration at indicated centres has been designated as "*R"
(first
eluted from the column in case the column conditions are described in the
synthesis
protocol and when only one stereocentre present) or "*S" (second eluted from
the
column in case the column conditions are described in the synthesis protocol
and when
only one stereocentre present) when the absolute stereochemistry is
undetermined
(even if the bonds are drawn stereospecifically) although the compound itself
has been
isolated as a single stereoisomer and is enantiomerically pure.
For example, it will be clear that compound 179
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>
N5
C)\:
F3C SN
is
R ____________________
N5
F3C or F3C
Compounds having two stereocentres of which only the stereochemical
configuration
.. of one stereocentre is indicated by * (e.g. *R or *S) (see for example
compound 186 or
281), follow a similar rule as above. This means that the absolute
stereoconfiguration of
the stereocentre indicated by * is undetermined (even if the bonds are drawn
stereospecifically) although the compound is enantiomerically pure at the
indicated
centre.
For compounds such as 188, 189, 190, 191, 235, 236, 237, and 238, wherein the
stereochemical configuration of two stereocentres is indicated by * (e.g. *R
or *S), the
absolute stereochemistry of the stereocentres is undetet mined (even if the
bonds are
drawn stereospecifically), although the compound itself has been isolated as a
single
stereoisomer and is enantiomerically pure. In this case, the configuration of
the first
stereocentre is independent of the configuration of the second stereocentre in
the same
compound.
For example, for Compound 188
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N )7-
C9N
N
/ _______ (n\ I
F3C S---N"---.
this means that the compound is
.¨.).: ....):
R
N/ N
N N
/ (n\
F3C S----..."-N---- F3C Sn----.N
",----
Or Or
N
R N-...._
---. / -
N/
N N
(n\ I
F3C Sn----.',N---- F3C S----..."-N----
Or .
The paragraphs above about stereochemical configurations, also apply to
intermediates.
The term "enantiomerically pure" as used herein means that the product
contains at
least 80% by weight of one enantiomer and 20% by weight or less of the other
enantiomer. Preferably the product contains at least 90% by weight of one
enantiomer
and 10% by weight or less of the other enantiomer. In the most preferred
embodiment
the term "enantiomerically pure" means that the composition contains at least
99% by
weight of one enantiomer and 1% or less of the other enantiomer.
When an intermediate or compound in the experimental part below is indicated
as 'HC1
salt', `FICOOH salt' or `TFA salt' without indication of the number of
equivalents of
HC1 or TFA, this means that the number of equivalents of HC1 or TFA was not
determined.
A skilled person will realize that, even where not mentioned explicitly in the
experimental protocols below, typically after a column chromatography
purification,
the desired fractions were collected and the solvent was evaporated.
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In case no stereochemistry is indicated in the spirocycle represented by Li,
this means
it is a mixture of stereoisomers, unless otherwise is indicated or is clear
from the
context.
When a stereo centre is indicated with 'RS' this means that a racemic mixture
was
obtained at the indicated centre, unless otherwise indicated.
A. PREPARATION OF THE INTERMEDIATES
0
N
i I )F3C
PREPARATION OF INTERMEDIATE l' :
A mixture of 4-chloro-6-(2,2,2-trifluoroethyl)thieno[2,3-d]pyrimidine (525 mg,
2.08
mmol) prepared as described in Journal of Medicinal Chemistry (2016), 59(3),
892-913,
tert-butyl 2,7-diazaspiro[4.5]decane-2-carboxylate (550 mg, 2.29 mmol) and
DIPEA
(1.43 mL, 8.3 mmol) in ACN (12 mL) was heated at 80 C overnight. The solution
was
cooled and the mixture was poured into cooled water, the product was extracted
with
Et0Ac, the organic layer was dried over MgSO4, filtered and evaporated to
dryness. The
residue was purified by chromatography over silica gel (stationary phase:
irregular 15-
40nm 50g, mobile phase: DCM/MeOH: gradient from 100/0 to 99/1). The product
containing fractions were collected and evaporated to dryness yielding 770 mg
(yield
81%) of intermediate 1.
The compounds in the Table below were prepared by using an analogous method as
described for the preparation of intermediate 1, starting from the respective
starting
materials
Intermediate number Structure Quantity (mg) Yield (%)
O)\ y___ 350 100
---o
Intermediate 2 (from 0
CAS[336191-17-4]
and [1628317-85-0]) C
N
/ 1
/
F3C S-----Nr
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Intermediate number Structure Quantity (mg) Yield (%)
200 73
0-4
Intermediate 3 (from Nt.....
CAS[885270-84-8]
and [1628317-85-0])
N
/ I 1
/
F3C S----N
00-......< 660 78
N
Intermediate 4 (from
CAS[885270-86-0]
and [1628317-85-0]) N
(------./L
/ I N
/
F3C S-----N
..õ...\ o
355 57
Intermediate 5 (from r.........740 (
CAS[885268-42-8]
LN
and intermediate 15)
(----XL
/ I N
F3C S Ni
. y
.
N 5
F3C/ S"----N
PREPARATION OF INTERMEDIATE 3:
A solution of 4-chloro-6-(2,2,2-trifluoroethyl)thieno[2,3-d]pyrimidine (11.4
g; 44.96
mmol), ter butyl 2,6 diazaspiro[3.4]octane-2-carboxylate (10.5 g; 49.46 mmol)
and
DIPEA (15.5 mL; 89.93 mmol) in iPrOH (183 mL) was heated at 90 C overnight.
The
solution was cooled to rt and the solution was poured into water then
extracted with
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Et0Ac (3X). The organic layer was washed with brine, dried over MgSO4 and
filtered
off.
A precipitate (in aqueous layer) was filtered off, washed with few DCM and
combined
with a previous filtrate. The solvent was evaporated to give 19.9 g of brown
solid. The
residue was taken up with diethylether, the precipitate was filtered and dried
to give 18.5
g of pale brown solid of intermediate 3 (96%).
Alternative preparation of intermediate 3:
To a mixture of 4-chloro-6- (2,2,2-trifluoroethyl)thieno[2,3-d]pyrimidine
(3.00 g, 11.9
mmol) and tert-butyl 2,6-diazaspiro[3.4] octane-2-carboxylate (2.5 g, 11.8
mmol) in
Et0H (50 mL) was added DIPEA (2 g, 15.5 mmol) in one portion. The mixture was
stirred at room temperature for 18 h. The mixture was evaporated and the
residue was
diluted in EA (200 mL). The solution was washed with water (100 mL*2), dried
over
Na2SO4, filtered and evaporated to give intermediate 3 (5.10 g, 11.9 mmol,
100% yield)
as brown oil.
----N
N H
/\....... .../
\ N/
/ I 1
/
F3C S-----N
PREPARATION OF INTERMEDIATE 6:
A mixture of intermediate 1 (770 mg, 1.69 mmol), and a solution of 4N HC1 in
dioxane
(4.22 mL, 16.9 mmol) in ACN (45 mL) was stirred at rt overnight. The mixture
was
poured out into iced water, basified with 3N NaOH, the product was extracted
with
DCM, the organic layer was dried over MgSO4, evaporated to dryness providing
670 mg
of intermediate 6 which was used without further purification for the next
step.
The compounds in the Table below were prepared by using an analogous method as
described for the preparation of intermediate 6, starting from the respective
starting
materials.
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Quantity Yield (%)
Intermediate number Structure
(mg)
H 320
Intermediate 7 (from )N
0
intermediate 2) N
F3C S "---- N
HC1 salt
H 582
N
Intermediate 8 (from
N
intermediate 4)
)
----- C
/ INj
/
F3C S"---N%
HC1 salt
>1:::C1
N
S
N
(, N
/
F3C S---N
PREPARATION OF INTERMEDIATE 9:
In a sealed tube, 4-chloro-6-(2,2,2-trifluoroethyl)thieno[2,3-d]pyrimidine
(0.6 g, 2.37
mmol) prepared as described in Journal of Medicinal Chemistry (2016), 59(3),
892-913,
tert-butyl 2,6-diazaspiro[3.3]heptane-2-carboxylate (0.57 g, 2.85 mmol), DIPEA
(0.82
mL, 4.75 mmol) in iPrOH(15 mL) were heated at 90 C for 2h. The solution was
cooled
to rt and the reaction mixture was poured into water then extracted with
Et0Ac. The
organic layer was washed with water, dried over MgSO4, filtered and evaporated
to
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dryness. The crude product was crystallized from Et20 providing 0.6 g (yield
61%) of
intermediate 9.
H
N
8
N
/ I 1F3C/ S
PREPARATION OF INTERMEDIATE 10:
A mixture of intermediate 9 (4.43 g; 10.69 mmol) in formic acid (24 mL) was
stirred at
RT overnight. The reaction mixture was evaporated. The residue was taken up
twice with
Et20 and evaporated to dryness. The residue was purified by chromatography
over silica
gel (irregular SiOH; 80 g; mobile phase: 90% DCM, 10% Me0H, 1% NH4OH). The
pure
fractions were collected and evaporated to dryness yielding 3.34 g (99%) of
intermediate
10.
H
N
8
N
/ I 1HCOOH salt
F3C
PREPARATION OF INTERMEDIATE 10b:
A mixture of intermediate 9 (0.55 g, 1.33 mmol) in formic acid (3 mL) was
stirred at RT
for 20 h. The mixture was evaporated in vacuo to give a residue that was taken-
up twice
with Et20 and evaporated to dryness giving 0.4 g (yield 96%) of intermediate
10b (formic
acid salt). The crude product was used without any further purification in the
next step.
The compounds in the Table below were prepared by using an analogous method as
described for the preparation of intermediate 10b, starting from the
respective starting
materials.
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Intermediate number Structure
----N
N H
/........ j
Intermediate 12 (from N/
intermediate 5)
/ IN
/
F3C S---N
HCOOH salt
H
\I
N
/ 1
S ----N
PREPARATION OF INTERMEDIATE 11: F3C/
A mixture of intermediate 3 (8.57 g; 20 mmol) in formic acid (51 mL) was
stirred at rt
for 20h. The reaction mixture was stirred at rt for the week-end. The mixture
was
evaporated and the residue was cooled to 5 C, taken-up with DCM and
neutralized with
aqueous solution of NaOH 3N. The organic layer was washed with water, dried
over
MgSO4, filtered and the solvent was evaporated. The residue (7.63 g of orange
oil) was
purified by chromatography over silica gel (irregular bare silica 120 g,
mobile phase: 1%
NH4OH, 85% DCM, 15% Me0H). The pure fractions were collected and the solvent
was
evaporated to give 3.65 g of yellow oil intermediate 11(56%).
Alternative preparation of intermediate 11:
TFA (17.9 mL; 233.38 mmol) was added to a solution of intermediate 3 (5 g;
11.67
mmol) in DCM (130 mL) at 5 C and the reaction mixture was stirred at rt for
4h. The
reaction mixture was diluted with heptane and evaporated to dryness (3X) to
give 10.7 g
of brown oil. The residue was purified by chromatography over silica gel
(irregular SiOH
40 gm; 220 g, mobile phase: 1% NH4OH, 90% DCM, 10% Me0H). The pure fractions
were collected and the solvent was evaporated. The residue (4 g) was
solubilized with
DCM and the product was crystallized. The mixture was evaporated and taken up
severals times with ACN and the solvent was evaporated to give 4 g of pale
yellow solid
intermediate 11.
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H
N
TFA salt
______________________________________ e\I
/
s-----
PREPARATION OF IN NTERMEDIATE 1 lb: F3
TFA (2.2 mL; 28 mmol) was added to a solution of intermediate 3 (600 mg; 1.4
mmol)
in DCM (13 mL) at 0 C then the reaction mixture was stirred at rt overnight.
The reaction
mixture was evaporated till dryness to give 1.26 g of intermediate 1 lb as TFA
salt. The
product was used it directly without purification.
H
c-3
N
HCI salt
(\j
/
F3c s--""N
PREPARATION OF INTERMEDIATE 1 lc:
A solution of HC1 4M in dioxane (150 mL) was added to intermediate 3 (6.5 g;
15.17
mmol) at rt. The reaction mixture was stirred at rt for lh. The mixture was
evaporated in
vacuum to give 5.7 g of yellow solid intermediate 11c as HC1 salt. The product
was used
without purification for the next step.
F
0
H N
\ /
H2N 1
0
PREPARATION OF INTERMEDIATE 13:
Acetic anhydride (1 mL, 10.7 mmol) was added dropwise at rt, to a solution of
2-amino-
5-(2,2,2-trifluoroethyl)thiophene-3-carboxamide (2 g, 8.92 mmol) in toluene
(50 mL)
and trimethylamine (6.2 mL, 44.6 mmol). The solution was heated at reflux for
5h,
poured into water, extracted with Et0Ac, and washed with brine (x2). The
organic layer
was dried over MgSO4, filtered and evaporated to dryness, the crude product
was taken-
up with Et20 and the precipitate was filtered to provide 1.5 g of intermediate
13 (yield
63% / brown solid).
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OH
/( ----) N
I
F3C S---
PREPARATION OF INTERMEDIATE 14:
To a solution of intermediate 13 (1.5 g, 5.63 mmol) in Et0H (70 mL) at rt, was
added
dropwise a 1M solution of KOH. The reaction mixture was stirred at rt for 30
min, then
the mixture was heated at reflux for 3h. The reaction mixture was cooled to rt
then poured
into ice water, acidified with 3N HC1, extracted with DCM and decanted. The
combined
organic layers were washed with brine and dried over MgSO4, filtered and
evaporated
to dryness. The residue was crystallized from Et20 to give 0.7 g of
intermediate 14 (yield
50%) that was used without further purification in the next step.
CI
/
/ I I
F3C S---
PREPARATION OF INTERMEDIATE 15:
Intermediate 14 (0.7 g, 2.82 mmol) and phosphoryl trichloride (5 mL) were
heated at 110
C for 2 h. The reaction mixture was cooled to rt, then evaporated to
dryness.The residue
was taken-up carefully with ice and DCM, basified with an aqueous solution of
K2CO3
(10%) and the organic layer was washed with water, dried over MgSO4, filtered
and
evaporated to dryness to give 0.75 g (yield 99%) of intermediate 15, that was
used
without further purification in the next step.
0¨s" '-
PREPARATION OF INTERMEDIATE 16: \
Spiro [3.3]heptan-2-ylmethyl methanesulfonate
To a solution of spiro[3.3]heptan-2-ylmethanol (153 mg, 1.08 mmol) in 4 mL of
DCM
was added TEA (0.464 mL, 3.2 mmol) and the reaction mixture was cooled to 0 C.
Methylsulfonylchloride (0.184 g, 1.605 mmol) was then added dropwise, the
mixture
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was allowed to warm to rt and stirred for 2h. An aqueous solution of saturated
NaHCO3
(30 mL) and DCM (30 mL) were added. The mixture was separated, the organic
layer
was collected, washed with brine (10 mL), dried over Na2SO4, and evaporated to
give
300 mg of intermediate 16 as a yellow oil which was used without further
purification in
the next step.
o
---Ao
N
1.) \
LbN
N
/ I
F3C/
S----
PREPARATION OF INTERMEDIATE 17: N
To a solution of intermediate 11c (400 mg) and TEA (0.38 mL, 2.76 mmol) under
a N2
flow in DCM (20mL) was added 1[2-(acetyloxy)ethy1]-1H-pyrrole-2-carboxaldehyde
(200 mg, 1.11 mmol). The mixture was stirred at rt for 4h. NaBH(OAc)3 (390 mg,
1.84
mmol) was added and the mixture was stirred at rt for 48h. Then, it was poured
into ice
water and the mixture was separated and the aqueous layer was extracted with
DCM.
The organic layers were combined, washed with brine then dried over MgSO4 and
evaporated to dryness. The residue was purified by chromatography over silica
gel
(stationary phase: irregular SiOH 15-40 m 24g, mobile phase: DCM/MeOH: 97/3).
The
pure fractions were collected and the solvent was evaporated under vacuum
yielding 180
mg of intermediate 17.
0......"
PREPARATION OF INTERMEDIATE 35
1H-pyrazole-4-carbaldehyde (0.5 g; 5.2 mmol) and cesium carbonate (3.39 g;
10.4
mmol) were diluted in ACN (10 mL). Then, 2-bromoethyl methyl ether (0.636 mL;
6.77
mmol) was added and the reaction mixture was refluxed for 2 hours. The
reaction mixture
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was partitionned between a saturated solution of NaHCO3 and Et0Ac. The organic
layer
was separated, dried over MgSO4, filtered and concentrated.
The residue was purified by silica gel chromatography (irregular SiO2, 120 g,
DCM/MeOH: 100/0 to 95/5). The fractions containing the product were mixed and
.. concentrated to afford 439 mg (55%) of intermediate 35.
-0
RS
0 LbN
/ I
F3C/
PREPARATION OF INTERMEDIATE 20:
Intermediate 11(150 mg, 0.46 mmol), (+/-)-methyl alpha-bromophenylacetate
(0.08
.. mL, 0.50 mmol) and K2CO3 (127 mg; 0.92 mmol) in DMF (10 mL) were stirred at
rt for
5h. The reaction mixture was poured into ice water and Et0Ac was added. The
organic
layer was separated, washed with brine, dried over MgSO4, filtered and
evaporated till
dryness. The residue was purified by chromatography over silica gel
(stationary phase:
irregular SiOH 15-40 m 24g, mobile phase: DCM/Me0H (+10% NH4OH): gradient
.. from 97/3 to 95/5). The pure fractions were collected and evaporated to
dryness yielding
162 mg (yield 74%) of intermediate 20.
HO
RS
O)
F3C
PREPARATION OF INTERMEDIATE 32
Lithium hydroxide monohydrate (71 mg; 1.7 mmol) was added, at rt, to a
solution of
intermediate 20 (162 mg; 0.34 mmol) in THF (3 mL) and water (3 mL). The
mixture was
.. stirred at rt overnight, then concentrated and acidified with an aqueous
solution of HC1
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3N (pH= 2-4).The precipitate was filtered and dry to give 33 mg (21%) of
intermediate
32 (90% of purity based on LC/MS). The mother layer was evaporated till
dryness to
give 243 mg of an impure fraction of intermediate 32.
0
0
(n\I
F3C
PREPARATION OF INTERMEDIATE 54 '
Intermediate 11c (333 mg), methyl 2-formylbenzoate (148.5mg; 0.905mmo1),
NaBH(OAc)3 (872mg; 4.11mmol) and trimethylamine (250mg; 2.47mmo1) were mixed
in dichloroethane (16 mL) and the reaction was stirred at RT overnight.Then,
an aqueous
solution of NaHCO3 (1(mL) was added and the mixture was extracted with DCM
(4*15
mL). The organic layers were separated, mixed, dried over MgSO4, filtered and
concentrated to afford 450mg of intermediate 54 as white solid.
The intermediates in the Table below were prepared by using an analogous
method as
described for the preparation of intermediate 54, starting from the respective
starting
materials.
Quantity Yield
Intermediate number Structure
(mg) (%)
= 0
340
\ro
Intermediate 44 (from No 0
intermediate 1 1 c and
intermediate 52)
/
F3C SN
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N
0
r ON
----tSD
N
N
______________________________________ / I
S
F 3C/
PREPARATION OF INTERMEDIATE 47
A mixture of intermediate 10b (150 mg), 1 -(tetrahydro -2H-pyran-2-y1)-1H-
pyrazo le-4-
carboxaldehyde (225 mg; 1.249 mmol) and AcOH (24 L; 0.416 mmol) in
dichloroethane (4.5 mL) was stirred at 50 C for 2 hours. The reaction mixture
was cooled
to room temperature and NaBH(OAc)3 (265 mg; 1.249 mmol) was added. The
reaction
mixture was stirred at room temperature overnight, poured onto a 10% aqueous
solution
of K2CO3 and extracted with DCM. The organic layer was decanted, dried over
MgSO4,
filtered and evaporated to dryness. The residue was purifed by chromatography
over
silica gel (irregular SiOH, 24g; mobile phase: gradient from 0% Me0H, 100% DCM
to
10% Me0H, 90% DCM). The pure fractions were collected and evaporated to
dryness
yielding 150 mg of intermediate 47.
The intermediates in the Table below were prepared by using an analogous
method as
described for the preparation of intermediate 47, starting from the respective
starting
materials.
Quantity Yield
Intermediate number Structure
(mg) (%)
rC¨Vo 158
N
Intermediate 48 (from N
intermediate 10b and 1-
8
(oxan-2-yl)pyrazo le-3-
N
carbaldehyde )
/ 1 NI,
/
F3C s---N
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N\
800
/ I N
F3C S N
PREPARATION OF INTERMEDIATE 25:
A mixture of (4 S)-1-Bo c-4-methyl-L-pro line(174 mg, 0.761 mmol), HBTU (288
mg,
0.761 mmol) and DIPEA (0.65 mL, 3.804 mmol) in DMF (7.5 mL) was stirred for
lh.
Then, a solution of intermediate 10 (250 mg, 0.761 mmol) in DMF (5 mL) was
added.
The reaction mixture was stirred at rt overnight. The reaction mixture was
poured into
iced water, basified with a 10% aqueous solution of K2CO3 and extracted with
Et0Ac.
The organic layer was washed with water, then brine, dried over MgSO4,
filtered and
evaporated to dryness. The residue (490 mg) was purified by chromatography
over silica
gel (irregular SiOH, 40g; mobile phase: NH4OH/DCM/MeOH: 0.5/95/5). The pure
fractions were collected and evaporated to dryness yielding 330 mg (yield 82%)
of
intermediate 25.
The intermediates in the Table below were prepared by using an analogous
method as
described for the preparation of intermediate 25, starting from the respective
starting
materials.
Quantity Yield
Intermediate number Structure
(mg) (%)
200
Intermediate 26 (from osso"s
intermediate 10b and N-Boc-
N
(2S,4S)-4-fluoropyrrolidine-
2-carboxylic acid)
/__C=jai N
I )
F3 C S
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Quantity Yield
Intermediate number Structure
(mg) (%)
....)...-0 0
\r 250 100
N 0
Intermediate 27 (from /
R
NE..7D
intermediate 11 and (R)-5-
Bo c azaspiro [2.4] heptane-6
carboxylic acid) N
/4-----N
i I )F3 C S N"....
0
\
8
N
O
N
/ (1)1
s------ F3C
PREPARATION OF INTERMEDIATE 104 N
DIPEA (0.48 mL; 2.775 mmol) was added to a solution of intermediate 10b (200
mg),
3-carboxybenzaldehyde (100 mg; 0.666 mmol) and HATU (317 mg; 0.833 mmol) in
DMF (10 mL) and the reaction mixture was stirred at room temperature for 4
hours. The
reaction mixture was poured onto water and extracted with Et0Ac. The organic
layer
was decanted, washed with water, then brine, dried over MgSO4, filtered and
evaporated
to dryness. The residue was purified by chromatography over silica gel
(irregular SiOH,
24g; mobile phase: gradient from 0% Me0H, 100% DCM to 10% Me0H, 90% DCM).
The pure fractions were collected and evaporated to dryness yielding 62 mg of
intermediate 104.
0
/
N",..õ..-",.
0 N ---- µ
oN
0
N
CriN
/
F3C S---\N:-
Preparation of intermediate 41:
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Under a N2 flow, at rt, to a solution of intermediate 11(250 mg, 0.76 mmol) in
DCM (12
mL) was added intermediate 42 (246 mg, 0.91 mmol). The mixture was stirred at
room
temperature for 3h. The mixture was cooled to 5 C, NaBH(OAc)3 (323 mg, 1.52
mmol)
was added and the mixture was stirred at rt overnight. Then, it was poured
into ice water
and the layers were separated. The aqueous layer was extracted with DCM. The
organic
layers were combined, washed with brine then dried over MgSO4, evaporated. The
residue was crystallized from Et20 and pentane. The white precipitate was
filtered off
and dried under vaccum yielding 55mg (yield 100%) of intermediate 41.
\r-
N
N
_____________________________________ / 1
/ S.---
F3C
Preparation of intermediate 43: N
Intermediate 11(500 mg, 1.52 mmol), 2-(chloromethyl)-1,1-dimethylethyl ester-
1H-
pyrro le-1-carboxylic acid) (493mg, 2.28mmo1) and K2CO3 (1.05g, 7.61mmo1) in
ACN
(12 mL) were stirred at room temperature for 24h. The reaction mixture was
poured into
ice water and Et0Ac was added. The organic layer was separated, washed with
brine,
.. dried over MgSO4, filtered and evaporated till dryness. The residue was
purified by
chromatography over silica gel (stationary phase: irregular SiOH 15-40 m 24g,
mobile
phase: NH4OH/DCM/MeOH: gradient from 0.1/97/3 to 0.1/95/5). The pure fractions
were mixed and evaporated yielding 100 mg (yield 14%) of intermediate 43.
The intermediates in the Table below were prepared by using an analogous
method as
.. described for the preparation of intermediate 43, starting from the
respective starting
materials.
Quantity Yield (%)
Intermediate number Structure
(mg)
ND H
157 66
/
Intermediate Si (from 1H- liv /
/ 0 With 60 C
pyrrole-4-carboxaldehydeand
as reaction
3 -bromopropionitrile) 1,1
temperature
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Quantity Yield (%)
Intermediate number Structure
(mg)
1000 71
Intermediate 42 (from 1H- 0
pyrrole-4-carboxaldehydeand N,7"-N = With reflux
as reaction
N-(2-bromoethyl) phthalimide) 0
temperature
340 99
Intermediate 52 (from 3-
With 75 C
hydroxybenzaldehyde and 3- 0 140
ONA)< as reaction
(Boc-amino)propyl bromide)
temperature
Intermediate 53 (from 1H- 1563 65
pyrazole-4-carbaldehydeand 2- With reflux
bromoethoxy-t-butyl ¨o as reaction
dimethylsilane) temperature
o __________________________________ (
N;lj
/
F3C
PREPARATION OF INTERMEDIATE 50:
Under a N2 flow, to a solution of intermediate 11(202 mg, 0.62 mmol) in DCM
(10 mL)
was added tert-butyl 4-formy1-1H-pyrazole-1-carboxylate (133 mg, 0.68 mmol)
and
AcOH (35 L, 0.62 mmol). The mixture was stirred at room temperature for 2h.
NaBH(OAc)3 (521 mg, 2.46 mmol) was added and the mixture was stirred at rt
overnight,
poured into ice water and the layers were separated. The aqueous layer was
extracted
with DCM. The organic layers were combined, washed with brine then dried over
MgSO4, evaporated. The residue was purified by chromatography over silica gel
(stationary phase: irregular SiOH 15-40 m 24g, mobile phase: DCM/Me0H(+10%
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NH4OH): 97/3). The pure fractions were mixed and evaporated yielding 145 mg
(yield
46%) of intermediate 50.
SIik
so
N
x_pl
RS
Ni.....
()
/4-----XLN
/ 1 1
F3 C S N(.7
PREPARATION OF INTERMEDIATE 55:
In a sealed tube, under a N2 flow, intermediate 53(349 mg, 1.37 mmol) and
Ti(OiPr)4
(436 L, 1.83 mmol) were added to a solution of intermediate 11(300 mg, 0.914
mmol)
in THF (6 mL). The solution was stirred at 50 C for 5 hours then, at rt
overnight. The
reaction mixture was cooled to 5 C and 2N iPrMgC1 in THF (2.28 mL, 4.57 mmol)
was
added dropwise. The reaction mixture was allowed to rise slowly to rt and
stirred
overnight. The reaction mixture was diluted with Et0Ac and poured onto a 10%
aqueous
solution of K2CO3. The insoluble material was removed by filtration over
celite . The
organic layer was decanted, washed with brine, dried over MgSO4, filtered and
evaporated to dryness. The residue was purified by chromatography over silica
gel
(irregular SiOH, 40g; mobile phase: Me0H/DCM: gradient from 0/100 to 10/90).
The
pure fractions were collected and evaporated to dryness yielding: 0.3 g (yield
54%) of
intermediate 55.
0
N
C\I
F3C/ S---"N
Preparation of intermediate 63 ,
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o
-1.-.
Ni *R ON40 (
63
N
F3 C/
S ----
intermediate 63a N-
o
N 0 (
63
N
Cj\:
F3 C/ S-----
and intermediate 63b N-
A solution of tert-butyl 3-acetylazetidine-1-carboxylate (364 mg; 1.83 mmol),
intermediate 11(400 mg; 1.22 mmol), titanium(IV)isopropoxide (725 L; 2.44
mmol)
in ethanol (2 mL) was stirred at 45 C for 30 min (solution become dark
yellow). Ethanol
(12 mL) and NaBH4 (138 mg; 3.66 mmol) were added and the solution become
yellow
pale. The reaction mixture was stirred at room temperature overnight. Then, it
was
poured onto a 10% aqueous solution of K2CO3 and DCM. The insoluble was
filtered
through a pad of celite . The organic layer was decanted, filtered through
chromabond
and the solvent was evaporated 624 mg of pale yellow oil which waspurified by
chromatography over silica gel (SiO2; 25 g; mobile phase: gradient from 98%
DCM, 2%
Me0H to 96% DCM, 4% Me0H). The fractions containing the product were collected
and the solvent was evaporated to give 223mg (36%) of intermediate 63 as a
white foam.
Intermediate 63 was purified by chiral SFC (Stationary phase: CHIRALCEL OJ-H
Sum
250x20mm, Mobile phase: 92% CO2, 8% Me0H(0.3% iPrNH2)). The pure fractions
were collected and the solvent was evaporated to give 83 mg (13%) of
intermediate 63a
as a colourless oil and 90 mg (14%) of intermediate 63b.
The intermediate in the Table below were prepared by using an analogous method
as
described for the preparation of intermediate 63, starting from the respective
starting
materials.
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Intermediate number Structure Quantity Yield
0
ci / 0 ( 600 mg 52%
Intermediate 60 (from N RSC\
intermediate 11 and
N-Boc-4-
acetylpiperidine F3c/
( N 4C) 221 mg 20%
*R 0 ( vi\N
Intermediate 60a and
intermediate 60b From
N
chiral SFC separation /
F3c S-
of intermediate 60:
(Chiralpak AD-H 5 intermediate 60a
gm 250*30 mm; ( \ ./c)
*s \ 0
mobile phase: 75%
CO2, 25% 229 mg 20%
iPrOH(0.3% iPrNH2)).
Cr,iN
F3c SN
Intermediate 60b
\ 0
N4
Intermediate 109 324 mg 33%0 ( RS
oN
(from intermediate 11
and intermediate 108) 0
F/ SN
Fr'F CN
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N
0
N RS c/ 0 (
F3C
PREPARATION OF INTERMEDIATE 68
\ _________________________________ '( \N
's _____________________________________ / 0 (
F3C
INTERMEDIATE 68a
N
0
0 (
F3C
INTERMEDIATE 68b and
c-3
e\j
s
COMPOUND 61: F3
A solution of intermediate 113 (1.67 g; 6.54 mmol) in THF (15 mL) was added to
a
solution of intermediate 11(1.4 g; 4.36 mmol) and TFA (2 mL; 26.16 mmol) in
THF (30
mL). The reaction mixture was stirred at rt overnight. Then NaBH(OAc)3 (2.77
g; 13.08
mmol) was added portionwise. The reaction mixture was stirred at rt for 10
days. The
solution was poured out into a 10% aqueous solution of K2CO3 and Et0Ac was
added.
The mixture was extracted with Et0Ac (3x). The organic layers were combined,
washed
with brine, dried over MgSO4, filtered and the solvent was evaporated. The
residue (2.9
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g; yellow oil) was purified by chromatography over silica gel (SiO2; 40 g;
eluent: from
97% DCM, 3% Me0H, 0.3% NH4OH to 90% DCM, 10% Me0H, 1% NH4OH). The
desired fractions were collected and the solvent was evaporated to give 266 mg
of
colourless oil intermediate 68, and 215 mg of colourless oil fraction 1.
Intermediate 68 was purified by chiral SFC (Lux-cellulose-2 5 gm 250*30 mm,
mobile
phase: 50% CO2, 50% Me0H (0.3% iPrNH2)). The pure fractions were collected and
the
solvent was evaporated to give 114 mg (5%) of colourless oil intermediate 68a
and 109
mg (4%) of colourless oil intermediate 68b.
Fraction 1 was purified by reverse phase (YMC-actus Triart C18 10 gm 30*150
mm,
mobile phase: gradient from 65% NH4HCO3 0.2%, 35% ACN to 25% NH4HCO3 0.2%,
75% ACN). The fractions containing the product were collected and the solvent
was
evaporated. The residue (160 mg; colourless oil) was freeze-dried with water-
ACN to
give 90 mg (6%) of white solid compound 61.
0
N/\OX
RS
N
(N11
F3C/ S"----e
PREPARATION OF INTERMEDIATE 69 ,
o
& ;1
NOX
N
/ _______________________ (N11
,,, F3C s-----
INTERMEDIATE Ova -
o
c9NoX
*S
N
,,,, F3C S----1\1
AND INTERMEDIATE 69b N
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A solution of 4-chloro-6-(2,2,2-trifluoroethyl)thieno[2,3-d]pyrimidine (5.07
g; 20.08
mmol), 2-B0C-2,7-diaza-spiro[4.4]nonane (5 g; 22.09 mmol) and DIPEA (6.9 mL;
40.17 mmol) in iPrOH (80 mL) was heated at 90 C overnight. The solution was
cooled
to rt and the solution was poured into water then extracted with Et0Ac (3X).
The organic
layer was washed with brine, dried over MgSO4 and the solvent was evaporated
to
dryness. The residue (9 g, pale brown solid) was taken up with diethylether,
the
precipitate was filtered and dried to give 8.4 g of intermediate 69 (95%, off-
white solid).
Intermediate 69 was purified by chiral SFC (Chiralpak IG 5 gm 250*20 mm,
mobile
phase: 65% CO2, 35% iPrOH (0.3% iPrNH2)). The pure fractions were collected
and the
solvent was evaporated to give 4.07 g of intermediate 69a (46%, yellow foam)
and 4.29
g of intermediate 69b (48%, yellow foam).
(NH
*R
(/
F3C SN
PREPARATION OF INTERMEDIATE 70a:
The intermediate 70a was prepared by using an analogous method as described
for the
alternative preparation of intermediate 11, starting from the respective
starting material
intermediate 69a.
The intermediate in the Table below were prepared by using an analogous method
as
described for the alternative preparation of intermediate 11, starting from
the respective
starting materials.
Intermediate number Structure Quantity Yield
H 3.4g Quant.
*s
Intermediate 70b (from
intermediate 69b
F3C SN
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BOC-N ___________________________________ s
NE...
(N)
/ I
F3C/
S
Preparation of intermediate 71 ,
BOC-N
*R N
lip
N
/
F3C s=-="N
intermediate 71a
\_
s
BOC¨ N
*S \
Ntb
N
F3C s-----N
and intermediate 71b
Method A:
A mixture of intermediate 1 lb (4.2 g) and intermediate 72 (1.6 g; 7 mmol) in
THF (50
mL) was stirred at RT overnight. Then, NaBH(OAc)3 (3 g; 14 mmol) was added
portion-wise. The reaction mixture was stirred at room temperature for 24
hours. The
solution was poured onto cold water, basified with an aqueous solution of NaOH
3N
and Et0Ac was added. The organic layer was separated, dried over MgSO4,
filtered
and evaporated to dryness. The residue was purified by chromatography over
silica gel
(irregular SiOH 80g, mobile phase: gradient from 98% DCM, 2% Me0H (+10%
NH4OH) to 95% DCM, 5% Me0H (+10% NH4OH)). The pure fractions were collected
and evaporated to dryness yielding 852 mg of intermediate 71.
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The enantiomers were separated by chiral SFC (CHIRALCEL OD-H 5gm 250*30mm;
mobile phase: 70% CO2, 30% Et0H). The pure fractions were collected and
evaporated
to dryness yielding 294 mg of intermediate 71a and 303 mg of intermediate 71b.
Method B:
The experiment was performed 6 times on the same quantity (640 mg; 1.95 mmol)
Ti(OEt)4 (0.8 mL; 3.9 mmol) was added at room temperature to a solution of
intermediate 11(640 mg; 1.95 mmol) and intermediate 72 (665 mg; 2.92 mmol) in
DCE (20 mL) and Me0H (8 mL). The reaction mixture was stirred at RT for 24h,
cooled at 10 C then NaBH3CN (367 mg; 5.84 mmol) was added portion wise. The
reaction mixture was stirred at room temperature for 8 days. The solutions
were
gathered for the work-up: poured out into cold water, basified with K2CO3
powder and
extracted with DCM. The suspension was filtered through a pad of Celite0. The
filtrate
was decanted, dried over MgSO4, filtered and evaporated to dryness.
The residue was purified by chromatography over silica gel (irregular SiOH,
40g;
mobile phase: gradient from 100% DCM, 0% Me0H to 97% DCM, 3% Me0H, 0.1%
NH4OH). The pure fractions were collected and evaporated to dryness yielding
1.7 g
(28%) of intermediate 71.
The enantiomers were separated by chiral SFC (Chiralcel OD-H 5gm 250*30mm;
mobile phase: 70% CO2, 30% Et0H (0.3% iPrNH2)). The pure fractions were
collected
and evaporated to dryness yielding 697 mg (11%) of intermediate 71a and 727 mg
(12%) of intermediate 71b.
The intermediates in the table below were prepared following Method A as
described
for the preparation of intermediate 71, 71a and 71b starting from the
respective starting
materials.
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Intermediate Structure Quantity Yield
number
C
intermediate 88 BO \N 130 mg
Li...SC
.....
N
CY-3
N
F3C/ / I
S-----N
From intermediate 1 lb and intermediate
89
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Intermediate Structure Quantity Yield
number
BOC
intermediate 82
NI 40 mg
11
F3I
From intermediate 1 lb and intermediate
83
BOO
NI
Intermediate 82a
Q
R* FR''
and
F3C
BOC
intermediate 82b S?F
(nN
F3I SN
From chiral SFC separation of
intermediate 82
(Stationary phase: CHIRALPAK AD-H
5ium 250*30mm, Mobile phase: 70%
CO2, 30% iP0H(0.3% iPrNH2))
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BOC
/
N
-(
11.1b
N
F C/
3 I \ / .. I
S ---- 1
PREPARATION OF INTERMEDIATE 77 ,
CB0
____________________ ( .-----'
_________________________ R
N
C\I
F3C/
intermediate 77a
BOC
/
N
( __________________________ p
_________________________ ,s N
1----ID
N
F3C S----N
and intermediate 77b
Reaction mixture 1: In a sealed tube, a solution of intermediate 78 (2
equivalents),
intermediate 11(100 mg; 0.305 mmol) and Ti(OiPr)4 (6 equivalents)in Et0H (0.2
mL)
was heated at 45 C for 1 hour. The mixture was cooled down to room
temperature,
diluted with Et0H (3 mL) and NaBH4 (2 equivalents) was added. The reaction
mixture
was stirred at room temperature for 4 hours indicating, according to LC/MS the
formation of 60% of intermediate 77
Reaction mixture 2 and 3: The reaction was performed twice on the same
quantity:
In a sealed tube, a solution of intermediate 78 (2 equivalents), intermediate
11 (450
mg; 1.37 mmol) and Ti(OiPr)4 (6 equivalents) in Et0H (0.9 mL) was heated at 45
C
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for 1 hour. The mixture was cooled down to room temperature, diluted with Et0H
(13
mL) and NaBH4 (2 equivalents) was added. The reaction mixture was stirred at
room
temperature for 18 hours.
The three reaction mixtures were diluted with Et0Ac and poured onto a mixture
of
10% K2CO3 and brine. The suspension was sonicated for 30 min and filtered
through a
pad of Celite . The organic layer was decanted, washed with 10% aqueous K2CO3,
then brine, dried over MgSO4, filtered and evaporated to dryness. The residue
(2.6g)
was purified by chromatography over silica gel (irregular SiOH, 50g; mobile
phase:
gradient from 0% NH4OH, 0% Me0H, 100% DCM to 0.7% NH4OH, 7% Me0H, 93%
DCM). The pure fractions were collected and evaporated to dryness. The residue
(1.5
g; 89%) was purifed a second time by chromatography over silica gel (irregular
SiOH,
40g; mobile phase: 60% Heptane, 35% Et0Ac, 5% Me0H (+10% NH4OH)). The
fractions containing the product were collected and evaporated to dryness
yielding 980
mg (58%; 82% pure besed on LC/MS) of intermediate 77.
The impure fraction of intermediate 77 was further purified by achiral SFC
(DIETHYLAMINOPROPYL 5 m 150x30mm; mobile phase: 90% CO2, 10% Me0H).
The pure fractions were collected and evaporated to dryness yielding 620 mg
(37%) of
intermediate 77.
The enantiomers were separated by chiral SFC (Lux Cellulose-2 5 m 250*30mm;
mobile phase: 50% CO2, 50% Me0H (0.3% iPrNH2)). The fractions containing the
products were collected and evaporated to dryness yielding 276 mg (16%) of
intermediate 77a and 269 mg (16%) of intermediate 77b.
The intermediates in the table below were prepared using an analogous method
as
described for the preparation of intermediate 77, 77a and 77b starting from
the
respective starting materials.
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Intermediate Structure Quantity Yield
number
intermediate 80 i CB0 240 mg 57%
N
/ r?
1----b
N
C?)2F3/
From
S-----\ N-
From intermediate 11 and tert-butyl 3-
propanoylazetidine-1-carboxylate
CN
0 0 __
Preparation of intermediate 72
Under N2 at 5 C, iPrMgC1 2M in THF (19 mL; 38.33 mmol) was added to a solution
of
intermediate 73 (4.6 g; 18.83 mmol) in THF (70 mL). The solution was stirred
at 5 C
for 30 min, allowed to slowly rise RT, stirred for lh then, heated at 40 C for
5h.
The reaction mixture was cooled to room temperature, poured out onto a mixture
of
iced water and a saturated aqueous NH4C1 solution, and extracted with Et0Ac.
The
organic layer was decanted, dried over MgSO4, filtered and evaporated to
dryness
yielding 4.7 g of intermediate 72 (quantitative).
The intermediates in the table below were prepared using an analogous method
as
described for the preparation of intermediate 72 starting from the respective
starting
materials.
intermediate Structure Quantity Yield
number
intermediate 75 / c =<,
_________________ N e 25 g Quant.
o o
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intermediate Structure Quantity Yield
number
From intermediate 73 and
cyclopropylmagnesium bromide 0.5 in
THF
intermediate 78 0 19g 96%
OA
N
1
)0n------
From intermediate 73 and
Isobutylmagnesium bromide 0.4M
intermediate 83 137 mg 49%
o
From intermediate 84 and
Isopropylmagnesium chloride 2M in
THF
intermediate 105 71 mg 16%
RSC0
From N-Methoxy-N-
methyltetrahydro furan-3 - carboxamide
and Isopropylmagnesium chloride 2M in
THF
Intermediate 108 0 710 mg 26%
ON 0
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intermediate Structure Quantity Yield
number
From N-Methyl-N-methoxy-1-
(tert-
butoxycarbonyppiperidine-4-acetamide
and Isopropylmagnesium chloride 2M in
THF
Intermediate 111 0 770 mg 40%
N
0 (
From intermediate 73 and chloro[(4-
methylphenyl)methyl]magnesium
Intermediate 113 1230 mg 33%
cN4
From 4-[(N-Methoxy-N-
methylamino)carbony1]-1-
piperidinecarboxylic acid 1,1-
dimethylethyl ester and
Isopropylmagnesium chloride 2M in
THF
\
-N 0
CN4o+
Preparation of intermediate 73
1-Boc-azetidine-3-carboxylic acid (5 g; 24.9 mmol) and N,0-
dimethylhydroxylamine
hydrochloride (3.64 g; 37.3 mmol) were placed in a round bottom flask under
N2. DCM
(75 mL) was added, followed by EDCI.HC1 (7.15 g; 37.3 mmol), DMAP (155 mg;
1.27
mmol) and DIPEA (6.5 mL, 37.4 mmol). The reaction mixture was stirred at RT
for 16
h and diluted with DCM (100 mL). The organic layer was washed with aqueous 1M
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HC1 (2 x 50 mL), sat. NaHCO3 solution (50 mL), and brine (50 mL). The organic
phase
was decanted, dried over MgSO4, filtered, and evaporated to dryness yielding
6.04 g
(99%) of intermediate 73.
The intermediates in the table below were prepared usingan analogous method as
described for the preparation of intermediate 73 starting from the respective
starting
materials.
intermediate Structure Quantity Yield
number
intermediate 84 0 F 600 mg 97%
o'N
I
From 1-Boc-3-fluoroazetidine-3-carboxylic
acid
o
/ N
\ 0
N
1 _________________________________
N3
L.-7HN
/ / 1
F3C S------N
Preparation of intermediate 85
Under N2, a solution of intermediate 11(204 mg; 0.62 mmol), intermediate 86
(217
mg; 0.81 mmol) and Ti(OEt)4 (0.26 mL; 1.24 mmol) in DCE (7 mL) was stirred at
RT
overnight. NaBH3CN (129 mg; 2 mmol) was added and the solution was stirred for
4
days. Water was added dropwise then the solution was filtered through a pad of
celite .
The filtrate was separated. The organic layer was washed with water, dried
over
MgSO4, filtered and evaporated till dryness. The residue was purified by
chromatography over silica gel (irregular SiOH, 24g; mobile phase: gradient
from 97%
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DCM, 3% Me0H (+10% NH4OH) to 95% DCM, 5% Me0H (+10% NH4OH)). The
pure fractions were collected and evaporated to dryness yielding 209 mg (80%)
of
intermediate 85.
The intermediate in the table below was prepared using an analogous method as
described for the preparation of intermediate 85 starting from the respective
starting
materials.
intermediate Structure Quantity Yield
number
intermediate 87 I 145 mg 51%
N
N
BOC N R
D S
N
Cni
F3/ S----N
From intermediate 11 and 4-[(1-methy1-1H-
pyrazol-4-y1)carbonyl]-1-
piperidinecarboxylic acid 1,1-dimethylethyl
ester
o
/ 0
N
0 ----- N
Preparation of intermediate 86
2-bromo-1-(1-methyl-1H-pyrazol-4-y1)-ethanone (0.5 g; 2.46 mmol) in DMF (10
mL)
were added to potassium phtalimide (0.46 g; 2.46 mmol). The reaction mixture
was
stirred at RT for 5h, poured into water-ice and Et0Ac was added. The organic
layer
was separated, washed with water, brine, dried over MgSO4, filtered and
evaporated till
dryness. The residue was purified by chromatography over silica gel (irregular
SiOH
15-40 m, 24g; mobile phase: 97% DCM, 3% Me0H (+10% NH4OH)). The pure
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fractions were collected and evaporated to dryness yielding 460 mg (69%) of
intermediate 86.
0
0/ N40-h
Preparation of intermediate 89
Under N2, n-BuLi 1.6M in hexane (6.2 mL; 9.92 mmol) was added at -70 C to a
solution of 4-iodo-1-methy1-1H-pyrazole (1.7 g; 8.17 mmol) in THF (35 mL). The
reaction mixture was stirred at -70 C for 1 hour then, a solution of
intermediate 73 (2 g;
8.19 mmol) in THF (10 mL) was added drop wise. The reaction mixture was
stirred at -
70 C for 2 hours, allowed to warm up to room temperature and stirred
overnight. The
solution was poured out into a mixture of ice-water and a saturated NH4C1
solution,
then Et0Ac was added. The organic layer was decanted, dried over MgSO4,
filtered
and evaporated to dryness. The residue was purified by chromatography over
silica gel
(irregular SiOH, 50g; mobile phase: gradient from 100% DCM, 0% Me0H to 98%
DCM, 2% Me0H, 0.1% NH4OH). The pure fractions were collected and evaporated to
dryness yielding 320 mg (15%) of intermediate 89
0
130C
eY\I
Preparation of intermediate 92 F3C
Under N2, HBTU (210 mg; 0.555 mmol) was added to a solution of BOC-L-proline
(;
119 mg; 0.555 mmol) and DIPEA (0.48 mL; 2.775 mmol) in DMF (10 mL). The
solution
was stirred for 30 min. Then, intermediate 10b (200 mg) was added and the
solution was
stirred at room temperature all over the weekend. Subsequently, the reaction
mixture was
poured into iced water, basified with a 10% aqueous solution of K2CO3 and
extracted
with Et0Ac. The organic layer was washed by water, dried over MgSO4, filtered
and
evaporated to dryness. The residue was purified by chromatography over silica
gel
(irregular SiOH, 24g; mobile phase: gradient from 0.5% NH4OH, 5% Me0H, 95% DCM
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to 1% NH4OH, 10% Me0H, 90% DCM). The pure fractions were collected and
evaporated to dryness yielding 168 mg of intermediate 92.
The intermediates in the table below were prepared using an analogous method
as
described for the preparation of intermediate 92 starting from the respective
starting
materials.
intermediate Structure Quantity Yield
number
intermediate 93 510 mg
0 N
" S
BOO
F3C/ SN
From intermediate 10b and (25,4R)-N-
boc-4-methylpyrrolidine-2-carboxylic
acid
intermediate 101 150 mg
0 .
.=== s N\
BOO
F30
From intermediate 10b and (2 S ,4R)-1-
(tert-butoxycarb ony1)-4-
fluoropyrro lidine-2-carboxylic acid
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intermediate Structure Quantity Yield
number
\
intermediate 96 0 283 mg 94%
0 . ----T;
N\130C
N
8
N
/ 1
F3C/ S----N%
From intermediate 10 and cis- 1 -N-Boc-
4-methoxy-L-proline
intermediate 102 200 mg
=== s N\
N BOO
8
N
Cni
/
F3C S----N"...-
From intermediate 10b and (S)-5-Boc-5-
Azaspiro[2.4]heptane-6-carboxylic acid
intermediate 103 ,\ io /1....
N N 160 mg
\
BOC 1...b
N
S----N-
F3C/ From intermediate 11c and (S)-5-(tert-
butoxycarbony1)-5-azaspiro[2.4]
heptane-6-carboxylic acid
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intermediate Structure Quantity Yield
number
intermediate 98 348 mg
BOC
Cni
F3C
From intermediate 10b and (R)-5-(tert-
butoxycarbony1)-5-azaspiro[2.4]
heptane-6-carboxylic acid
Intermediate 99 0 389 mg 75%
s
0 ..
BOC
F3C
From intermediate 10 and (2S,4R)-1-
(tert-butoxycarbony1)-4-
methoxypyrrolidine-2-carboxylic acid
Intermediate 100 100 mg
0 S
BOO
F3C
From intermediate 10b and N-B0C-4,4-
difluoro-L-proline
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) 0
N
_____________________________________________ / 1
F3C/
PREPARATION OF INTERMEDIATE 107
Under nitrogen, a solution of intermediate 11 (250mg; 0.76 lmmol), N-Boc-4-
formylpiperidine (195mg; 0.914mmo1) in THF (7mL) was stirred at rt for 3h.
NaBH(OAc)3 (323mg; 1.52mmo1) was added and the mixture was stirred at rt
overnight.
A 10% aqueous solution of K2CO3 and DCM were added. The organic layer was
separated, dried over MgSO4, filtered and evaporated to dryness. The residue
was
purified by silica gel chromatography (irregular SiOH, 24g; mobile phase:
gradient from
100% DCM 0% Me0H (0%NH4OH) to 90% DCM 10% Me0H (10%NH4OH)). The
fractions containing the product were mixed and evaporated to dryness yielding
383mg
(96%) of intermediate 107.
0 I 0
s*
I I
0
F
PREPARATION OF INTERMEDIATE 110:
DIPEA (0.45 mL, 3.24 mmol) was added to an ice-cooled solution of 2-(4-
fluorophenyl)
propanol (CAS[59667-20-8]) (0.25 g, 1.62 mmol) in DCM (1.4 mL) followed by
methane sulfonyl chloride (0.155 mL, 1.95 mmol). The mixture was stirred
overnight at
rt. The mixture was diluted with DCM (20 mL) and washed with a saturated
sodium
bicarbonate solution (15 mL). The solution was dried over MgSO4, filtered and
concentrated under reduced pressure yielding 0.377 g of intermediate 110 .
This product
was used without further purification in the next step.
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o y
o
N
RS
4. Nt....
cl)
/ I
F3C/ S----N-
PREPARATION OF INTERMEDIATE 112:
A solution of intermediate 111(607 mg, 2.1 mmol) and titanium isopropoxyde
(1.25
mL, 1.37 mmol) in Et0H (4.6 mL) was added dropwise at room temperature (over a
period of 5 to 10 min) to a mixture of intermediate 11 (459 mg, 1.4 mmol) and
NaBH3CN (264 mg, 4.2 mmol) in Et0H (9.2 mL). The mixture was stirred for 1 h
at rt.
The reaction mixture was diluted with DCM and poured onto a 10% aqueous
solution
of K2CO3. The suspension was filtered over a pad of Celite0. The organic layer
was
decanted, washed with brine, dried over MgSO4, filtered and evaporated to
dryness.
The residue was purified by chromatography over silica gel (irregular SiOH, 24
g;
.. mobile phase: DCM/MeOH: gradient from 100/0 to 90/10). The pure fractions
were
collected and evaporated to dryness yielding 0.521g (62%) of intermediate 112
(62%).
B. PREPARATION OF THE COMPOUNDS
EXAMPLE B1
1.1
-N
I
)
N
eN
/
F3C S-
PREPARATION OF COMPOUND 1:
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TEA (88.5 mg, 0.875 mmol) and benzaldehyde (46.4 mg, 0.44 mmol) were
successively
added to a solution of intermediate 11(190 mg, 0.44 mmol) in anhydrous DCM (4
mL)
and the mixture was stirred at rt for 30 min. NaBH(OAc)3 was then added (185.4
mg,
0.875 mmol) and the mixture was stirred at rt overnight. Sat. aq. NaHCO3 (10
mL) and
DCM (10 mL) were added and the mixture decanted. The aqueous layer was
extracted
twice with DCM (10 mL). The organic layers were combined, washed with brine
(10
mL), dried over Na2SO4 and evaporated to give a yellow oil. The crude residue
was
purified by chromatography over silica gel (column Gemini 150*25 Sum, mobile
phase:
water (0.05% ammonia hydroxide v/v)/ACN: gradient from 55/45 to 25/75). The
residue
was then freeze-dried to give 65 mg of compound 1 (35% yield) as a yellow
solid.
F F
F-\
\I"----bN
N
/ I 1
/
F3C S-----N%
EXAMPLE B2
PREPARATION OF COMPOUND 2:
Intermediate 11 (100 mg, 0.3 mmol), 3,3,3-trifluoropropanal (51 mg, 0.46 mmol)
in dry
DCM (3 mL) were stirred at rt for lh, then NaBH(OAc)3 (129 mg, 0.61 mmol) was
added
and the mixture was stirred at rt overnight. The mixture was poured into water
then
extracted with DCM, the organic layer was dried over MgSO4, filtered and
evaporated
to dryness. The residue was purified by chromatography over silica gel
(stationary phase:
irregular SiOH 15-40 m 24g, mobile phase: NH4OH/DCM/MeOH: 0.1/97/3). The
product containing fractions were collected and evaporated to dryness yielding
58 mg
(45%) of compound 2, which was freeze-dried with ACN/water 20/80 to give 45 mg
of
compound 2.
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-----
Ntb
N
F,C S"..-N
Preparation of compound 13
A mixture of intermediate 11c (600 mg), isobutyraldehyde (160 mg; 2.221 mmol),
NaBH(OAc)3 (1.57 g; 7.405 mmol) and Et3N (0.64 mL; 4.443 mmol) in DCE (12 mL)
was stirred at room temperature overnight. A saturated aqueous solution of
NaHCO3 (20
mL) and DCM (20 mL) were added. The organic layer was decanted and the aqueous
layer was extracted with DCM (20 mL*2). The combined organic layers were
washed
with brine (30 mL), dried over Na2SO4, filtered and evaporated to dryness. The
residue
was purified by chromatography over silica gel (mobile phase: gradient from
petroleum
ether/Et0Ac from 100/0 to 0/100, then Et0Ac/ Me0H from 100/0 to 85/15). The
pure
fractions were collected, evaporated to dryness and freeze dried yielding 320
mg of
compound 13.
The compounds in the Table below were prepared by using an analogous method as
described for the preparation of compound 2, starting from the respective
starting
materials.
Quantity Yield (%)
Compound number Structure
(mg)
. 80
_-N
Compound 3 (from
intermediate 7) N/
es--LN
F3 C/ S , I j
-----N
as a hydrochloride salt
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Quantity Yield (%)
Compound number Structure
(mg)
N*I\I 126 47
Compound 4 (from -N
intermediate 11) I
)
N
es---LN
/
F3C S-N)
75 43
_II\I
Compound 5 (from
N
intermediate 11)
1-----b
N
/
C----)N / I
,
F3C S N
55 26
riN
Si
Compound 6 (from
N
intermediate 11)
1----b
N
C----)N
/ / I
F3C S N
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Quantity Yield (%)
Compound number Structure
(mg)
55 31
(1\--1).2
Compound 7 (from (N
intermediate 11)
1------b
N
C---)N
/ / I
F3C S-N
CI 75 43
¨N
Compound 8 (from
N
intermediate 11)
1----7-3
N
/ I 1
/
F3C Si\i%
N 75 39
2
Compound 9 (from N
intermediate 11)
1---b
N
C----LN
/ / I
F3C S.---N
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Quantity Yield (%)
Compound number Structure
(mg)
N---. 40 29
2
Compound 10 (from N
intermediate 11)
1-b
N
C----LN
/ / I
F3C S'N
65 37
¨N
Compound 11 (from N
intermediate 11)
1---7-3
N
/ / I Il
F3C
p 80 41
¨N
Compound 12 (from N
intermediate 11)
1-b
N
/
C----LN / I
,
F3C S'N
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Quantity Yield (%)
Compound number Structure
(mg)
---- 80 54
b
Compound 13 (from ,
intermediate 11)
N
C-----)N
/ / I
F3C S---N
-N /N 70 18
__,..j
Compound 23 (from Nicb
intermediate 11 and 1-
methy1-1H-imidazo le-
N
5-carboxaldehyde )
(nNi
F1 s------N,
ki 155 40
NU
Compound 35 (from
o
intermediate 11 and N
1-
methyl-1H-pyrazo le-
0
3-carbaldehyde ) N
F3C S ---- \
111)0 60 22
Compound 69 (from
intermediate 11 and
\¨]
phenylacetaldehyde
(CAS[122-78-1])) 0
N
1
F3C // S----N
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oN
O
N
F
F Cr\I
S N
EXAMPLE B3 F
PREPARATION OF COMPOUND 14:
Intermediate 10b (0.42 g), benzyl bromide (0.19 mL, 1.6 mmol), and K2CO3 (0.55
g, 4.0
mmol) in ACN (20 mL) were stirred at rt overnight. The mixture was poured into
water,
extracted with Et0Ac, the organic layer was washed with brine, then dried over
MgSO4,
filtered and evaporated to dryness. The residue was purified by chromatography
over
silica gel (stationary phase: irregular SiOH 15-40 m 40g, mobile phase:
NH4OH/DCM/MeOH: 0.1/97/3). The product containing fractions were collected and
evaporated to dryness yielding 170 mg (31%) of compound 14, which was
crystallized
from DIPE, filtered and dried to give 103 mg of compound 14.
The compounds and intermediates in the Table below were prepared by using an
analogous method as described for the preparation of compound 14, starting
from the
respective starting materials.
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Quantity Yield (%)
Compound number Structure
(mg)
N 65 37
-N
Compound 15 (from N
intermediate 11)
1---7-3
N
/
HN / I
,
F3C S.---N
57
N
Compound 16 (from
intermediate 8) N
C N"--)
/ / I j
F3C S'N
as a hydrochloride salt
(1.7HC1. 1.9H20)
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Quantity Yield (%)
Compound number Structure
(mg)
N 60 32
11
/ NH
Compound 17 (from
intermediate 11)
4111P
(int. 11 was reacted
with 1628318-10-4,
N
followed by cleavage
1---7
with TFA)
¨3
N
(I 1
/
F3C S---"N
N
4* 130 49
c.i
Compound 18 (from y)
intermediate 6) N
/4-XLN
)F3C Q =-= N
---\ 45 12
Compound 19 (from
intermediate 12)
/ 1 1F3 C S N''''
N
_31 28 13
Compound 30 (from
LbN
intermediate 11 and
chloromethyltrimethyl
N
germane)
e--iN
,
Fõ s_¨,N,
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Quantity Yield (%)
Compound number Structure
(mg)
/ Ge 36 21
,...
(
Compound 31 (from oN
intermediate 11 and
0 chloromethyltrimethyl N
germane)
/
F3C S----\ N---ij
EXAMPLE B4
N
C---h
N
/ I 1\1
/
F3C
PREPARATION OF COMPOUND 20:
A mixture of 4-chloro-6-(2,2,2-trifluoroethyl)thieno[2,3-d]pyrimidine (150 mg,
0.59
mmol) prepared as described in Journal of Medicinal Chemistry (2016), 59(3),
892-913,
2-benzyl 2,7-diaza-spiro-[4.4]nonane (CAS[885275-27-4]) (129 mg, 0.59 mmol)
and
DIPEA (0.31 mL, 1.78 mmol) in ACN (15 mL) were heated at 80 C overnight. The
mixture was cooled and poured into cooled water, the product was extracted
with Et0Ac,
the organic layer was dried over MgSO4, filtered and evaporated to dryness.
The residue
was purified by chromatography over silica gel (stationary phase: irregular
bare silica 24
g, mobile phase: DCM/Me0H/NH4OH: 97/3/0.1). The product containing fractions
were
collected and evaporated to dryness yielding 200 mg (yield 78%) of compound 20
(racemic mixture).
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2
. 4fk
N
.R
N) N
*S"
N
C-----)N HN
F3C __________________________________ S----N F3C __ S----N%
Preparation of enantiomers 20a and 20b
The enantiomers were separated via chiral SFC (stationary phase: Lux Cellulose-
4 5 m
250*21.2mm, mobile phase: CO2/Me0H (0.3% iPrNH2): 70/30). The product
containing
fractions were collected and evaporated to dryness yielding 80 mg (yield 31%)
of a first
eluted fraction Fl and 81 mg (yield 31%) of a second eluted fraction F2.
Fl (80 mg; 0.185 mmol) was dissolved in acetone, at 10 C, and 4N HC1 in
dioxane (2
eq, 0.37 mmol, 934) was added followed by Et20. The mixture was evaporated to
dryness and taken up with Et20, a precipitate was filtered and dried giving 65
mg (yield
20%) of compound 20a as a hydrochloride salt (1.95HC1 . 1.25H20 . 0.19 Dioxane
. 0.06
.. Et20)
F2 (81 mg, 0.187 mmol) was dissolved in acetone, at 10 C, and 4N HC1 in
dioxane (2
eq, 0.37 mmol, 93 L) was added followed by Et20. The mixture was evaporated
to
dryness, taken up with Et20, a precipitate was filtered and dried giving 49 mg
(yield
15%) of compound 20b as a hydrochloride salt (2.0HC1 . 1.8H20).
EXAMPLE B5
=
----N
N
/\....... .../
N
/C s=-)N
/ I I
F3C _________________________________ S--"N
PREPARATION OF COMPOUND 18:
A mixture of intermediate 6 (222 mg, 0.63 mmol), benzyl bromide (82 4, 0.685
mmol)
and K2CO3 (430 mg, 3.11 mmol) in ACN (20 mL) was stirred at rt overnight. The
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solution was poured out into cooled water, the product was extracted with
Et0Ac, the
organic layer was dried over MgSO4, filtered and evaporated to dryness. The
residue was
purified by chromatography over silica gel (stationary phase: irregular 15-40
gm 30 g,
mobile phase: DCM/Me0H/NH4OH: gradient from 100/0/0 to 97/3/0.1). The product
containing fractions were collected and evaporated to dryness yielding 170 mg
(yield
61%) of compound 18 (racemic mixture).
N
_____________________________________________ / I
F3C
Preparation of enantiomers compound 21a
=
"S "gil
\N/
/ I
F3C SN
And compound 21b
Compound 18 was separated into its enantiomers via chiral SFC (stationary
phase: Lux
Cellulose-2 5gm 250*30mm, mobile phase: CO2/MeOH: 75/25). The product
containing
fractions were collected and evaporated to dryness yielding 72 mg (yield 26%)
of a first
eluted fraction Fl and 76 mg (yield 27%) of a second eluted fraction F2.
Fl was dissolved in acetone (3 mL), a solution of 4N HC1 in dioxane (2eq, 80
gL, 0.32
mmol) was added dropwise at 10 C, Et20 was added and after 30 min a
precipitate was
filtered and dried giving 54 mg (yield 16%) of compound 21a as a hydrochloride
salt
(1.8HC1 . 1.9H20).
F2 was dissolved in acetone (3 mL), a solution of 4N HC1 in dioxane (2eq, 85
gL, 0.34
mmol) was added dropwise at 10 C, Et20 was added and after 30 min a
precipitate was
filtered and dried giving 34 mg (yield 10%) of compound 21b as a hydrochloride
salt
(1.8HC1 . 2.1H20).
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EXAMPLE B6
1--111.
N
1---7-3
N
/ I 1
/
F3C S--"N
PREPARATION OF COMPOUND 22:
To a solution of intermediate 11 (200 mg, 0.51 mmol) in ACN (5 mL) was added
intermediate 16 (220 mg, 1.08 mmol) and K2CO3 (221.4 mg, 1.53 mmol). The
mixture
was heated to 90 C and stirred overnight. Water (10 mL) and DCM (10 mL) were
added
to the reaction mixture. The organic phase was separated, the aqueous layer
was extracted
with DCM (10 mL). The organic layers were combined, washed with brine (10 mL),
evaporated to give a residue which was purified by chromatography over silica
gel
(Column: Gemini 150*25 5u; mobile phase: water (0.05% ammonia hydroxide
v/v)/CH3CN: gradient from 42/58 to 12/88, gradient Time (min): 10; 100% B Hold
Time
(min): 2; flow Rate (ml/min): 25).
The desired fractions were collected and dried in vacuum to give the residue.
The residue
was lyophilized to give 65 mg (yield 28%) of compound 22 as a light yellow
solid.
EXAMPLE B7
___/----N '
HO
NEb
N
F3C S----N
PREPARATION OF COMPOUND 24:
Intermediate 17 (180 mg, 0.36 mmol) and 3N NaOH (0.61 mL, 1.82 mmol) in Me0H
(10 mL) were stirred at rt for 1 h. The mixture was cooled to rt, poured into
water and
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extracted with Et0Ac. The organic layer was dried over MgSO4, filtered and
evaporated
to dryness. The residue was freeze-dried with acetonitrile/water 20/80
yielding 130 mg
of compound 24(79% yield).
EXAMPLE B8
NH
(i1:11
N
F3C/
S----\ N
PREPARATION OF COMPOUND 25:
At 5 C, to a solution of intermediate 43 (100 mg, 0.2 mmol) in DCM (10 mL), 4N
HC1
in dioxane (246 L, 0.99 mmol) was added dropwise and the mixture was stirred
at rt for
15h. The reaction was evaporated to dryness. Then, the residue was taken-up
with DCM,
washed with NaHCO3. The organic layer was dried over MgSO4, filtered and
evaporated
to dryness. The residue was purified by chromatography over silica gel
(stationary phase:
irregular SiOH 15-40 m 24g, mobile phase: NH4OH/DCM/Me0H gradient from
0.5/95/5 to 1/90/10). The pure fractions were collected and evaporated to
dryness. The
residue was freeze-dried with acetonitrile/water 20/80 yielding 25 mg of
compound
25(31% yield).
The compounds in the Table below were prepared by using an analogous method as
described for the preparation of compound 25, starting from the respective
starting
materials.
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Quantity Yield
Compound number Structure
(mg) (%)
H2N
250 84
=
Compound 37 (from
intermediate 44) LbN
/ N
F3C S
N 0 35 17
Compound 112 (from
1-1-3
intermediate 27)
/ I N
F3C SN
EXAMPLE B9
101
/ I N
F3C
PREPARATION OF COMPOUND 26:
In a sealed tube, 4-chloro-6-(2,2,2-trifluoroethyl)thieno[2,3-d]pyrimidine
(0.15 g, 0.594
mmol), 6-(phenylmethoxy)-2-azaspiro[3.3]heptane (0.145 g, 0.713 mmol) and
DIPEA
(0.205 mL, 1.19 mmol) in isopropanol (2 mL) were heated at 90 C overnight. The
solution was cooled to rt and poured into water then extracted with Et0Ac. The
organic
layer was washed with water, dried over MgSO4, filtered and evaporated to
dryness. The
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crude product was crystallized from Et20 and dried. The residue was purified
by
chromatography over silica gel (15-40 gm, 24 g, eluent: heptane/Et0Ae: 80/20
to 20/80).
The pure fractions were mixed and the solvent was evaporated. The residue was
taken
up by Et20, filtered and dried yielding 0.111 g of compound 26 (45% yield).
EXAMPLE B10
rC)N-
.1 C4H404
_____________________________________ / I N
F3C/
S
PREPARATION OF COMPOUND 28:
A mixture of intermediate 10b (200 mg), 1-methyl-1H-pyrazole-4-carboxaldehyde
(183
mg; 1.66 mmol) and AcOH (32 gL; 0.555 mmol) in DCE (6 mL) was stirred at 50 C
for 2 hours. The reaction mixture was cooled to room temperature and
NaBH(OAc)3
(353 mg; 1.665 mmol) was added. The reaction mixture was stirred at room
temperature overnight, poured onto a 10% aqueous solution of K2CO3 and
extracted
with DCM. The organic layer was decanted, dried over MgSO4, filtered and
evaporated
to dryness. The residue was purifed by chromatography over silica gel
(irregular SiOH,
24g; mobile phase: gradient from 0% NH4OH, 0% Me0H, 100% DCM to 1% NH4OH,
10% Me0H, 90% DCM). The pure fractions were collected and evaporated to
dryness
yielding 165 mg of compound 28 as an oil (73%). Compound 28 was dissolved in
ACN and HC1 (4N in dioxane) (277 gL; 1.11 mmol) was added. The HC1 salt was
filtered but revealed to be too hydroscopic. The residue was then dissolved in
DCM/Me0H and the organic layer was washed with a 10% aqueous solution of
K2CO3, dried over MgSO4, filtered and evaporated to dryness. The resulting
residue
was dissolved in ACN and fumaric acid (47 mg; 0.404 mmol; 1 eq) was added and
the
solution was allowed to stand until crystallization (overnight). The
precipitate was
filtered, washed with ACN then Et20 and dried yielding 188 mg of compound 28
as the
fumarate salt (1 equivalent based on 1H NMR).
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The compounds in the Table below were prepared by using an analogous method as
described for the preparation of compound 28, starting from the respective
starting
materials.
Quantity Yield
Compound number Structure
(mg) (%)
/ 1
N¨N 47
(11,)
Compound 34 (from intermediate N
10b and 1-methyl-1H-pyrazo le-3-
S
carboxaldehyde ) N
F3C
.......
Compound 44 (from intermediate N
10b and isobutyraldehyde )
8
N
Fl
S---N
5 EXAMPLE B11
H
N
NiC..
dr\31
N
C---)N
/ I
,, /
, F3C S'....-)
PREPARATION OF COMPOUND .v: N
At 10 C, 4N HC1 in dioxane (0.7 mL; 2.85 mmol) was added to a solution of
intermediate 50 (145 mg; 0.28 mmol) in ACN (7mL). The solution was stirred at
rt
overnight. The solution was evaporated to dryness. The residue was taken in
ice water,
10 basified with NH4OH and DCM was added. The organic layer was separated,
dried
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over MgSO4, filtered and evaporated till dryness. The residue was purified by
chromatography over silica gel (stationary phase: irregular SiOH 15-40 m 12g,
mobile
phase: DCM/Me0H/NH4OH 90/10/10). The pure fractions were collected and
evaporated to dryness. The residue was freeze-dried with acetonitrile/water
20/80
yielding 0.050 g (43% yield) of compound 29.
The compounds in the Table below were prepared by using an analogous method as
described for the preparation of compound 29, starting from the respective
starting
materials.
Quantity Yield (%)
Compound number Structure
(mg)
H
N 102 61
1\10
N
Compound 40 (from 8
N
intermediate 47)
/ 1
F3c/ s----N
2.68HC12.1H20
H
N¨N 72 55
c_k
N
Compound 41 (from
8
intermediate 48
N
/ 1
F3C/
S----N
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Quantity Yield (%)
Compound number Structure
(mg)
0
70 43
1
ys
Compound 88 (from
intermediate 26
F3C/ / I
EXAMPLE B12
NN
(N)
/ I
F3C/
PREPARATION OF COMPOUND 36:
Under a N2 flow, to a solution of intermediate 11(287 mg, 0.87 mmol) in DCM
(14
mL) was added 1-isopropyl-1H-pyrazole-4-carbaldehyde (133 mg, 0.68 mmol) and
AcOH (51 L, 0.87 mmol). The mixture was stirred at rt for 2h. NaBH(OAc)3 (742
mg,
3.5 mmol) was added and the mixture was stirred at rt overnight. The mixture
was
poured into ice water and was separated. The aqueous layer was extracted with
DCM.
The organic layer was washed with brine then, dried over MgSO4, filtered and
evaporated. The residue was purified by chromatography over silica gel
(stationary
phase: irregular SiOH 15-40 m 24g, mobile phase: DCM/Me0H (+10% NH4OH):
gradient from 97/3 to 90/10. The pure fractions were collected and evaporated
to
dryness. The residue was freeze-dried with acetonitrile/water: 20/80 yielding
0.057 g
(15% yield) of compound 36.
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The compounds in the Table below were prepared by using an analogous method as
described for the preparation of compound 36, starting from the respective
starting
materials.
Quantity Yield (%)
Compound number Structure
(mg)
N
73 33
N.._õ,
/...._'N
Compound 76 (from
intermediate 11 and
N
intermediate 51)
1-----7¨)
N
/ 1 1
F3C S N
EXAMPLE B13
NLb
N
/ I
/ S----N
PREPARATION OF COMPOUND 45 F3
To a solution of intermediate 11c (200mg) in dichloroethane (10 mL) was added
2-
methylbenzaldehyde (59 mg; 0.494mmo1), NaBH(OAc)3 (523mg; 2.47mmo1) and
triethylamaine (150mg; 1.48mm01). The mixture was stirred at room temperature
overnight and then, a saturated aqueous solution of NaHCO3 (10 mL) and DCM (10
mL)
were added.The mixture was separated and the aqueous layer was extracted with
DCM
(10 mL*2).
The organic layers were combined, washed with water (10 mL), dried over
Na2SO4,
evaporated to give 300mg of a yellow oil which was purified by preparative
high-
performance liquid chromatography (Column: Kromasil 150*25mm*10um; Conditions:
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A: water (0.05% ammonia hydroxide v/v), B: MeCN, at the beginning: A (52%) and
B
(48%), at the end: A: (22%) and B (78%), Gradient Time(min) 8;; Flow
Rate(ml/min)
30.
The fractions containing the product were collected and the solvent was
evaporated under
vacuum.The aqueous layer was lyophilized to dryness to give 150mg (70%) of
compound 45 as white solid.
The compounds in the Table below were prepared by using an analogous method as
described for the preparation of compound 45, starting from the respective
starting
materials.
Quantity Yield (%)
Compound number Structure
(mg)
190
II
Compound 46 (from
.)
intermediate 11c and m-
7.....
to lualdehyde )
N
/ / I
F3C S---"\ 1,1
- HC1 salt
\o
120 .
Compound 47 (from
Ni....b
intermediate 11c and 2-
methoxy-5-
N
methylbenzaldehyde )
/ 1 1
F3c S c N".7
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Quantity Yield (%)
Compound number Structure
(mg)
F 150
F .
Compound 48 (from
intermediate 11c and 2,4- Ni.......)
difluorobenzaldehyde )
N
/ 1 N
F3C
160
gi
Compound 49 (from No
intermediate 11c and p-
tolualdehyde ) 0
N
/ 1
F3C S-----N
155
11
Compound 50(from
intermediate 11c and 2,4- b
dimethylbenzaldehyde )
0
N
F3C s*----NN
HC1 salt
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Quantity Yield (%)
Compound number Structure
(mg)
166
F *
Compound 54(from
Ni.b
intermediate 11c and 2-
fluorobenzaldehyde )
N
C-XLN
/ I I
,
F3C/ 0 N
HC1 salt
F
150
411
Compound 56(from
intermediate 11c and 3- NL...13
fluorobenzaldehyde )
N
/ e------ry
F3 s---- --
1\1 HCl salt
EXAMPLE B14
N
8
N
/ I
F3C/
S-----N
PREPARATION OF COMPOUND 108
A mixture of S)-5 -methyl-5 -az aspiro [2 .4] heptane-6-carboxylic acid (94
mg; 0.61 mmol),
HBTU (231 mg; 0.61 mmol) and DIPEA (0.52 mL; 3.04 mmol) in DMF (5 mL) was
stirred for 1 hour. Then, a solution of intermediate 10b (200 mg) in DMF (5
mL) was
added and the reaction mixture was stirred at room temperature overnight. The
reaction
mixture was poured into iced water, basified with a 10% aqueous solution of
K2CO3 and
extracted with Et0Ac. The organic layer was washed by H20, then brine, dried
over
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MgSO4, filtered and evaporated to dryness. The residue was purified by
chromatography
over silica gel (irregular SiOH, 10g; mobile phase: gradient from 3% Me0H, 97%
DCM
to 10% Me0H, 90% DCM). The fractions containing the product were collected and
evaporated to dryness yielding 144 mg of an impure fraction 1. A second
purification
was performed (irregular SiOH, 40g; mobile phase: 0.5% NH4OH, 95% DCM, 5%
Me0H). The fractions containing the product were collected and evaporated to
dryness
yielding 43 mg of an impure fraction 2.
Fraction 2 was purified again by chromatography over silica gel (irregular
SiOH, 10g;
mobile phase: gradient from 3% Me0H, 97% DCM to 10% Me0H, 90% DCM). The
fractions containing the product were collected and evaporated to dryness. The
resulting
residue was taken up with diisopropyl ether. The solid was filtered and dried
yielding 17
mg of compound 108.
The compounds in the Table below were prepared by using an analogous method as
described for the preparation of compound 108, starting from the respective
starting
materials.
Quantity Yield (%)
Compound number Structure
(mg)
NH 10 7
Compound 74 (from H N
0
intermediate 32 and N- . RS
ip
isopropylethylenediami Ill
ne )
N
I_C---XLN
1 I )
F3C S N"...
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Quantity Yield (%)
Compound number Structure
(mg)
OH 22 13
HN
Compound 75 (from
= intermediate 32 and 2- RS
aminoethanol )
/ I
F3C SN
EXAMPLE B18
HO 40
HCI salt
NLb
/ I
F3C N
PREPARATION OF COMPOUND 57:
LiA1H4 (66 mg, 1.73 mmol) was added to intermediate 54 (450 mg, 0.693 mmol) in
THF
(12 mL). The reaction was stirred at rt for 1.5h. The reaction was quenched
with a
saturated aqueous solution of NH4C1, extracted with DCM and concentrated to
afford a
white solid. This solid was purified by preparative high-performance liquid
chromatography (column: Xtimate C18 150*25mm*10um, condition: water (0.05%
ammonia hydroxide,v/v)/ACN: gradient from 52/48 to 42/58). To the aqueous
layer was
added 0.1 mL 1N HC1. The solution was freeze-dried yielding 30 mg of compound
57 as
a yellow solid (HC1 salt).
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EXAMPLE B19
)------\ HCI salt
N
/ I
F3C/ S"---N
PREPARATION OF COMPOUND 58:
To a solution of intermediate 8 (200 mg, 0.5 mmol) in THF (10 mL) was added
isobutyraldehyde (70 L, 0.77 mmol) and TEA (0.37 mL, 2.63 mmol). The mixture
was
stirred at rt for 3h. NaBH(OAc)3 (317 mg, 1.5 mmol) was added and the solution
was
stirred at rt overnight. The solution was poured out into cooled water and was
basified
with K2CO3 powder. The product was extracted with Et0Ac. The organic layer was
dried
over MgSO4, filtered and evaporated to dryness. The residue was purified by
chromatography over silica gel (stationary phase: irregular bare silica 40g,
mobile phase:
NH4OH/DCM/MeOH: 0.2/98/2). The residue was dissolved in 5 mL of ACN, 2eq of 4N
HC1 in dioxane (117 L; 0.47 mmol) was added dropwise at 10 C. Et20 was added
and
after 30mn, the solution was evaporated to dryness, Et20 was added and a
precipitate
was filtered and dried yielding 38 mg of compound 58 (HC1 salt).
The compounds in the Table below were prepared by using an analogous method as
described for the preparation of compound 58, starting from the respective
starting
materials.
Quantity Yield (%)
Compound number Structure
(mg)
N
p 73
Compound 59 (from (¨)N
intermediate 8 and 1-
methy1-1H-pyrazo le- 6
N
4-carbaldehyde )
F3 C S N
HC1 salt
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EXAMPLE B21
0 H
N,
) _____________________________________ RS
N
lip
N
/ I 1F3C/
S'"--N
PREPARATION OF COMPOUND 62:
1M TBAF in THF (0.815 mL, 0.815 mmol) was added dropwise to a solution of
intermediate 55 (0.248 g, 0.407 mmol) in Me-THF (8 mL) and the reaction
mixture was
stirred at rt overnight. The reaction mixture was poured onto a 10% aqueous
solution of
K2CO3 and extracted with Et0Ac. The organic layer was washed with 10% aqueous
K2CO3 (30 mL), water (30 mL) and brine (30 mL), dried over MgSO4, filtered and
evaporated to dryness. The residue was purified by chromatography over silica
gel (80
g, 15-40 gm, eluent: DCM/MeOH: 97/3 to 88/12). The pure fractions were mixed
and
the solvent was evaporated yielding 0.043 g of compound 62 (21% yield).
EXAMPLE B22
vp
\ ¨Rs
c..N.31
()
/1 N
F3C/ S----N
PREPARATION OF COMPOUND 63:
2,2-Dimethyl-tetrahydropyran-4-carbaldehyde (87 mg; 0.609 mmol) and NaBH(OAc)3
(645 mg; 3.045 mmol) were added at rt to a solution of intermediate 11(200 mg;
0.609
mmol) in DCE (4 mL) and the reaction mixture was stirred overnight. The
reaction
mixture was diluted with DCM and poured onto a 10% aqueous solution of K2CO3.
The
organic layer was decanted, dried over MgSO4, filtered and evaporated to
dryness. The
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residue was purified by chromatography over silica gel (irregular SiOH, 24g;
mobile
phase: NH4OH/Me0H/DCM: gradient from 0/0/100 to 0.7/7/93). The pure fractions
were collected and evaporated to dryness. The residue was freeze-dried from
water/ACN
(80/20; 10 mL) yielding 155 mg of compound 63 (56% yield).
The compounds in the Table below were prepared by using an analogous method as
described for the preparation of compound 63, starting from the respective
starting
materials.
Quantity Yield (%)
Compound number Structure
(mg)
c_o 71 60
Compound 65 (from (
NL..b
intermediate 11 and
3 -methylo xetane-3-
N
carbaldehyde )
/ e------y
s---N
F3C
EXAMPLE B23
F
.........F
F
( --1
N
/ I 1
F3C S----\ N
PREPARATION OF COMPOUND 64:
A mixture of intermediate 11 (150 mg, 0.457 mmol), 2,2,2-trifluoroethyl
trifluoromethanesulfonate (69 L, 0.502 mmol) and DBU (CAS[6674-22-2]) (136
L,
0.914 mmol) in DMSO (3 mL) was stirred at rt for 18h. The reaction mixture was
poured
onto water and extracted with Et0Ac. The organic layer was washed several
times with
water, then brine, dried over MgSO4, filtered and evaporated to dryness. The
residue was
purified by chromatography over silica gel (irregular SiOH, 24g; mobile phase:
NH4OH/Me0H/DCM gradient from 0/0/100 to 0.7/7/93). The pure fractions were
collected and evaporated to dryness. The residue was purified a second time by
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chromatography over silica gel (irregular SiOH, 10g; mobile phase:
Et0Ac/heptane:
gradient from 60/40 to 80/20). The pure fractions were collected and
evaporated to
dryness. The residue was crystallized from DIPE yielding, after drying under
vacuum at
50 C, 100 mg of compound 64 (53% yield).
EXAMPLE B24
F.)
,
F3/ __________________________________ s,,N)
PREPARATION OF COMPOUND 67:
Under N2, to a solution of intermediate 11 (100 mg; 0.31 mmol), 2-(tetrahydro-
2H-
pyran-4-y1))acetaldehyde (48 L; 0.37 mmol) in THF (3 mL) were stirred at rt
for 3h.
NaBH(OAc)3 (129 mg; 0.61 mmol) was added and the mixture was stirred at rt
overnight . A 10% aqueous solution of K2CO3 and Et0Ac were added. The mixture
was
extracted with Et0Ac (x3).The organic layers were combined, washed with brine
then
dried over MgSO4, filtered and the solvent was evaporated.
The residue (136mg) was purified by chromatography over silica gel (SiO2, 4g;
gradient:
from 95% DCM, 5% Me0H, 0.5% NH4OH to 90% DCM, 10% Me0H, 1% NH4OH ).
The fractions containing the product were collected and the solvent was
evaporated to
give 90 mg of colourless oil which was recrystallized with diisopropylether.
The
precipitate was filtered and dried to give 45 mg (34%) of compound 67 as a
white solid.
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EXAMPLE B25
o
N
\ .:IN
RS
LbN
N
F3C / I."- S
PREPARATION OF COMPOUND 71 N
In a sealed tube, under N2, intermediate 35 (211 mg; 1.37 mmol) and Ti(OiPr)4
(4364;
1.83 mmol) were added to a solution of intermediate 11 (300 mg; 0.914 mmol) in
THF
(6 mL). The solution was stirred at 50 C for 5 hours then at rt overnight. The
reaction
mixture was cooled to 5 C and isopropyl magnesium chloride 2M in THF (2.28 mL;
4.57
mmol) was added dropwise. The reaction mixture was allowed to rise slowly to
rt and
stirred overnight. The reaction mixture was diluted with Et0Ac and poured onto
a 10%
aqueous solution of K2CO3. The precipitate was removed by filtration over
Celite . The
organic layer was decanted, washed with brine, dried over MgSO4, filtered and
evaporated to dryness. The residue was purified by chromatography over silica
gel
(irregular SiOH, 40g; mobile phase: gradient from 0% Me0H, 100% DCM to 10%
Me0H, 90% DCM). The fractions containing the product were collected and
evaporated
to dryness to give 0.337 g of an intermediate residue which was purified again
by
chromatography via reverse phase (stationary phase: YMC-actus Triart C18 101m
30*150mm, mobile phase: gradient from 55% NH4HCO3 0.2%, 45% ACN to 0%
NH4HCO3 0.2%, 100% ACN). The pure fractions were collected and evaporated to
dryness. The residue was freeze-dried with acetonitrile/water 20/80 to afford
120 mg
(26%) of compound 71.
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EXAMPLE B26
(NH2
N
J-N1
HCI salt
/ I
F3C
PREPARATION OF COMPOUND 77:
Hydrazine (36 L, 0.92 mmol) was added to a solution of intermediate 41(110
mg, 0.18
mmol) in ethanol (5 mL). The solution was heated at 70 C for 1h30. The
reaction was
cooled to rt, then poured into water and extracted with DCM. The organic layer
was
washed with brine, then dried over MgSO4, filtered and evaporated to dryness.
The
residue was purified by chromatography over silica gel (stationary phase:
irregular bare
silica 40g, mobile phase: NH4OH/DCM/MeOH: 1/85/15). The pure fractions were
collected and evaporated to dryness. The residue was dissolved in 2 mL of ACN,
3 eq of
6N HC1 in iPrOH were added dropwise at 10 C. Et20 was added and after 30mn,
the
precipitate was filtered and dried yielding 83 mg of compound 77(37% yield).
EXAMPLE B28
o..6
/ I
F3C
PREPARATION OF COMPOUND 62:
TFA (1.5 mL) was added to a solution of intermediate 25 (300 mg, 0.571 mmol)
in DCM
(15 mL) and the reaction mixture was stirred for 18h. The reaction mixture was
poured
onto a 10% aqueous solution of K2CO3 and extracted with DCM. The organic layer
was
decanted, filtered over chromabond and evaporated to dryness. The residue was
purified
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by chromatography over silica gel (irregular SiOH, 10g; mobile phase:
NH4OH/Me0H/DCM: gradient from 0.3/3/97 to 1.5/15/85). The pure fractions were
collected and evaporated to dryness. The residue was crystallized from DIPE
and dried
yielding 118 mg of compound 82 (48% yield).
The compounds in the Table below were prepared by using an analogous method as
described for the preparation of compound 82, starting from the respective
starting
materials.
Quantity Yield (%)
Compound number Structure
(mg)
)
140 52
S
( )N H
N
Compound 315
1-----b
(from intermediate
109) N
C\I
F
S------N
F
F
H 200
N
Compound 318 RS
(from intermediate 11 N
112 and T = 0 C)
N
TFA salt
HN
/ I j
/
F3C S----N
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EXAMPLE B29
F
fi
RS
ILlip
N
/ 1 y
F3C/
S
PREPARATION OF COMPOUND 84:
Intermediate 1 lb (198 mg) and 4-fluorophenylacetone (68 L, 0.51 mmol) in THF
(5
mL) were stirred at rt overnight. Then NaBH(OAc)3 (161 mg, 0.76 mmol) was
added
portionwise. The mixture was stirred at rt for 24h. The solution was poured
out into
cooled water and basified with a solution of 3N NaOH, Et0Ac was added. The
organic
layer was separated, dried over MgSO4, filtered and evaporated to dryness. The
residue
was purified by chromatography over silica gel (stationary phase: irregular
bare silica
40g, mobile phase: NH4OH/DCM/MeOH: 0.1/97/3). The pure fractions were
collected
and the solvent was evaporated under vacuum. The residue was freeze-dried with
acetonitrile/water 20/80 yielding 30 mg of compound 84.
The compounds in the Table below were prepared by using an analogous method as
described for the preparation of compound 84, starting from the respective
starting
materials.
Quantity Yield
(%)
Compound number Structure
(mg)
F
46
fi
Compound 307 RS
Nib
(from intermediate 1 lb
and 1-(4-fluoropheny1)-3-
N
metylbutan-2-one)
/ 1 nil
F3c s
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EXAMPLE B32
N
ijN RS
, õ F3C/ SN
PREPARATION OF COMPOUND
N
\
*R
N
33
(n\
C S COMPOUND 137 N
_N
\
N
C)\
F3C/
and COMPOUND 138
Ti(OEt)4 (2514; 1.2mmo1) was added at room temperature to a solution of
intermediate
11 (200mg; 0.6mmo1) and 2,5,6,7-tetrahydro-2-methy1-4H-Indazo1-4-one (120mg;
0.8mm01) in dichloroethane (5mL) and Me0H (1.5mL). The mixture was stirred at
rt for
2 hours then NaBH3CN (127mg; 2mm01) was added portionwise. The mixture was
.. stirred at room temperature for 2 days. The solution was poured out into
cooled water
and DCM was added. The mixture was basified with K2CO3 powder, filtered
through a
pad of celite . The product was extracted with DCM. The organic layer was
dried over
MgSO4, filtered and evaporated to dryness.
The residue (361 mg) was purified by silica gel chromatography (Stationary
phase:
irregular bare silica 40g, Mobile phase: 0.5% NH4OH, 95% DCM, 5% Me0H). The
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fraction containing the product were mixed and concentrated to afford 120mg
(43%) of
compound 133.
Chiral separation of compound 133 was performed via chiral SFC (Stationary
phase:
Chiralpak AD-H 5 m 250*30mm , Mobile phase: 70% CO2, 30% Me0H(0.3%
iPrNH2)). The fractions containing the products were mixed and concentrated to
afford:
- 45mg of fraction 1 which was freeze-dried with acetonitrile/water 20/80
to give
40mg (43%) of compound 137 as a white powder.
- 46mg of fraction 2 which was freeze-dried with acetonitrile/water 20/80
to give
42mg (46%) of compound 138 as a white powder.
EXAMPLE B33
NRS 0
63
N
/ I
S-----\ N
, F3C/
PREPARATION OF COMPOUND 145 ,
6.3N *R C
N
/ I
F3 C/ S"----N
COMPOUND 154
0
oN
N)
CL)\1
F3C/ SN
AND COMPOUND 155
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A solution of 1-tetrahydro-2H-pyran-4-ylethanone (351 mg; 2.74 mmol),
intermediate
11(600 mg; 1.83 mmol), Ti(OiPr)4 (870 L; 2.92 mmol) in Et0H (3 mL) was
stirred for
2 hours at 45 C. Additional Et0H (18 mL) and NaBH4 (138 mg; 3.65 mmol) were
added.
The reaction mixture was stirred at room temperature for 5 hours. The reaction
mixture
was diluted with DCM and poured onto a 10% aqueous solution of K2CO3. The
insoluble
material was removed by filtration over celite . The organic layer was
separated, washed
with water, dried over MgSO4, filtered and evaporated to dryness. The residue
was
purified by chromatography over silica gel (irregular SiOH, 24g; mobile phase
: gradient
from 0% NH4OH, 0% Me0H, 100% DCM to 1% NH4OH, 10% Me0H, 90% DCM).
The fractions containing the product were collected and evaporated to dryness
yielding
487 mg (60%) of compound 145. The enantiomers of compound 145 were separated
by
chiral SFC (CHIRALPAK AD-H 5 m 250*30mm; mobile phase: 70% CO2, 30%
mixture of Et0H/iPrOH 50/50 v/v). The fractions containing the products were
collected
and evaporated to dryness. The residues were freeze dried from from water/ACN
(80/20;
10 mL) yielding 171 mg (21%) of compound 154 and 178 mg (22%) of compound 155.
The compounds in the Table below were prepared by using an analogous method as
described for the preparation of compound 145, starting from the respective
starting
materials. The most relevant minor deviations are indicated in the column
'Compound
number'.
Compound number Structure Quantity Yield
Compound 279 (from
( I 243 mg 37%
c.1Rs o
intermediate 11 and
1 -(3 -methylo xetan-3 -
(N5
yl)ethenone [1363381-
04-7]
e--)
/
With 6eq. of Ti(iPrO)4 F3o S -------N
and 2eq. of NaBH4.
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Compound number Structure Quantity Yield
77 mg 12%
aN)*R LI
0
N
Compound 158 and F3 C /
S
compound 159 From
SFC purification of
Compound 158
intermediate
158CHIRACEL OJ-H 5
(
gm 250*30 mm, mobile
phase: 80% CO2, 20%
80 mg 12%
Et0H(0.3% iPrNH2))
F3c/
SN
Compound 159
14%
Compound 192 (from ( 76 mg
intermediate 11 and 1-
(tetrahydro-2,6-
N5
dimethy1-2H-pyran-4-
y1)-1-Propanone
With 1.6eq. of Ti(iPrO)4 F3
SN
and 2eq. of NaBH4
195 mg
Compound 280 (from
N
intermediate 1 lb and 3-
RS
methyl-1-(tetrahydro -
2H-pyran-4-y1)-1-
Butanone With 6eq. of
Ti(iPrO)4 and 2eq. of F3
NaBH4
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Compound number Structure Quantity Yield
70 mg 10%
/0
Compound 179 and
compound 180 From /
F30 s---Nr
SFC purification of
compound Compound 179
280CHIRALPAK AD-
H 5 gm 250*30 mm;
69 mg 10%
mobile phase: 75% CO2,
25% Me0H(0.3%
iPrNH2))
Cf)2F3C/ S
Compound 180
Compound 250
,o 369 mg 62%
With 6eq. of Ti(iPrO)4
LbN
and 2eq. of NaBH4
/ I N
F3C
Compound 250
From intermediate 11 and 1-
(1,1-dioxidotetrahydro-2H-
thiopyran-4-yl)ethenone
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Compound number Structure Quantity Yield
o
\\ o
s%
compound 163
) 123 mg 21%
1.....bN
and
N
e\j
F3C/ S----N1
Compound 163
compound 164
0
\\ 0
) 123 mg 21%
---.s
D
N
Cn\I
F31 S----N
Compound 164
From compound 250 chiral
SFC: CHIRACEL OJ-H
5gm 250*30mm; mobile
phase: 70% CO2, 30%
Me0H (0.3% iPrNH2)
N
Compound 314 // 105 mg 15%
(From intermediate 11
and 4-
Cyanophenylacetone
RS
ciN
N5
e--r
F3c/ s---,N_
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PREPARATION OF COMPOUND 312 (Diastereoisomer A (mixture of 2 compounds (RR
and SS) or (RS and SR)) and
COMPOUND 313 (Diastereoisomer B (mixture of 2 compounds (RS and SR) or (RR and
SS)):
Co 312: Diastereoisomer A (RR and SS) or (RS and SR)
Co 313: Diastereoisomer B (RS and SR) or (RR and SS)
1 0
( i Nil H
N
/ 1 1
F3c/ s--"N
Reaction mixture 1: A solution of 3-methyl-1-(6-oxaspiro[4.5]dec-9-y1)-1-
Butanone
(1.5 eq.), intermediate 11(100 mg; 0.285 mmol), Ti(OiPr)4 (1.6 eq.) in ethanol
(0.25
mL) was stirred for 2 hours at 45 C. Ethanol (3 mL) was added and NaBH4 (2
eq.) was
added. The reaction mixture was stirred at room temperature for 18 hours.
Reaction mixture 2 : A solution of 3-methyl-1-(6-oxaspiro[4.5]dec-9-y1)-1-
Butanone
(546 mg; 2.436 mmol; 2eq), intermediate 11(400 mg; 1.22 mmol), Ti(OiPr)4 (580
L;
1.95 mmol) in ethanol (1 mL) was stirred for 2 hours at 45 C. Ethanol (12 mL)
was added
and NaBH4 (92 mg; 2.436 mmol) was added. The reaction mixture was stirred at
room
temperature for 18 hours.
The two reaction mixtures were gathered and diluted with Et0Ac, poured onto a
10%
aqueous solution of K2CO3 and filtered through a pad of Celite . The organic
layer was
decanted, washed with brine, dried over MgSO4, filtered and evaporated to
dryness. The
residue was purified by chromatography over silica gel (irregular SiOH, 24g;
mobile
phase: gradient from 0% NH4OH, 0% Me0H, 100% DCM to 0.9% NH4OH, 9% Me0H,
91% DCM). The desired fractions were collected and evaporated to dryness.
The residue (280 mg) was further separated by chromatography over silica gel
(irregular
SiOH, 24g; mobile phase: 67% heptane, 33% Et0Ac (+ 5% Me0H containing 10%
NH4OH)). The desired fractions were collected and evaporated to dryness
yielding:
- 100mg (12%) of compound 312 (eluted first; not pure enough) which was
further
purified by chromatography over silica gel (irregular SiOH, 24g; mobile phase:
gradient
from 0% NH4OH, 0% Me0H, 100% DCM to 0.5% NH4OH, 5% Me0H, 95% DCM).
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The pure fractions were collected and evaporated to dryness. The pure
fractions were
freeze dried from water/ACN (80/20; 10 mL) yielding 80 mg (10%) of compound
312.
- 110mg (13%) of compound 313 (eluted second; not pure enough) which was
further
purified by chromatography over silica gel (irregular SiOH, 24g; mobile phase:
gradient
from 0% NH4OH, 0% Me0H, 100% DCM to 0.5% NH4OH, 5% Me0H, 95% DCM).
The pure fractions were collected and evaporated to dryness. The pure
fractions were
freeze dried from water/ACN (80/20; 10 mL) yielding 70 mg (9%) of compound
313.
EXAMPLE B34
\ NH
N/ *R ______________________________________________ /
N
C*---)N
/ I j
/
F3C S----N
PREPARATION OF COMPOUND 147:
A mixture of intermediate 60a (216 mg; 0.4 mmol) and TFA (1 mL; 13.067 mmol)
in
DCM (10 mL) was stirred at rt for 4h. The reaction mixture was diluted with
DCM and
basified with a 10% aqueous solution of K2CO3. The organic layer was decanted,
washed
with water, filtered through Chromabond0 and evaporated to dryness. The
residue (200
mg) was purified by chromatography over silica gel (irregular SiOH, 10 g;
mobile phase:
85% DCM, 14% Me0H, 1% NH4OH). The pure fractions were collected and evaporated
to dryness. The residue was freeze-dried from water/ACN (80/20; 10 mL)
yielding 136
mg of compound 147 (77%).
The compounds in the Table below were prepared by using an analogous method as
described for the preparation of compound 147, starting from the respective
starting
materials.
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Compound number Structure Quantity Yield
130 mg 71%
CN H
Compound 146 (from
intermediate 60b
C)\1
F3C
N H 29 mg 45%
N/ 'R
Compound 150 (fromN5
intermediate 63a
F3c s
50 mg
CN H
RS
Compound 148 (from
intermediate 68
TFA salt
F3/ C)\I
SN
45 mg 50%
*R( NH
Compound 168 (from
intermediate 68a (N)-
Cn\I
F3C S
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Compound number Structure Quantity Yield
*s CN H 50 mg 58%
Compound 169 (from
intermediate 68b (N
/ (TN
F3C S ----N--.-
EXAMPLE B35
RS
N 0
&N)
e--riN
,
F3c _______________________________ s¨"--N-,
PREPARATION OF COMPOUND 156
c3N/ -IR ___________________________ /
&N)
e¨nN
,
F3c __________________ S ----"",N
COMPOUND 193 ,
*s co
N __
&N ----3
e¨nN
,
F3c __________________ S ----"",N
COMPOUND 162
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RS 0
&N)-
, F3C
AND COMPOUND 157:
A solution o f 2-methyl-1 -(tetrahydro -2H-pyran-4-y1)-1 -Prop anone (1.66 g;
10.63 mmol)
in THF (30 mL) was added to a solution of intermediate 11(2.33 g; 7.08 mmol)
and TFA
(3.3 mL; 42.5 mmol) in THF (45 mL). The reaction mixture was stirred at rt
overnight.
Then NaBH(OAc)3 (4.5 g; 21.25 mmol) was added portionwise. The reaction
mixture
was stirred at rt for 7 days. The reaction mixture was stirred at rt for 3
days. The solution
was poured out into a 10% aqueous solution of K2CO3, Et0Ac was added. The
mixture
was extracted with Et0Ac (3x). The organics layers were combined, washed with
brine,
dried over MgSO4, filtered and the solvent was evaporated. The residue (3.9 g;
yellow
oil) was purified by chromatography over silica gel (SiO2; 40 g; eluent: from
97% DCM,
3% Me0H, 0.3% NH4OH to 90% DCM, 10% Me0H, 1% NH4OH). The pure fractions
were collected and the solvent was evaporated to give 666 mg of pale brown
solid
compound 156.
Compound 156 was purified by reverse phase (YMC-actus Triart C18 10 gm 30*150
.. mm, mobile phase: gradient from 50% NH4HCO3 0.2%, 50% ACN to 0% NH4HCO3
0.2%, 100% ACN). The pure fractions were collected and the solvent was
evaporated to
give 66 mg of compound 157 (colourless oil) and 264 mg of compound 156 (8%;
colourless oil).
50 mg of compound 156 was freeze-dried with water-ACN to give 47 mg of
compound
156 ( white solid).
Compound 157 was freeze-dried with water-ACN to give 53 mg of compound 157
(2%,
white solid).
Compound 156 (214 mg) was purified by chiral SFC (CHIRALPAK AD-H 5 gm 250*30
mm, mobile phase: 75% CO2, 25% Et0H (0.3% iPrNH2)). The pure fractions were
collected and the solvent was evaporated to give 82 mg of compound 193
(colourless oil)
and 82 mg of compound 162 ( colourless oil).
Compound 193 was freeze-dried with water-ACN to give 72 mg of compound 193
(2%,
white solid).
Compound 162 was freeze-dried with water-ACN to give 77 mg of compound 162
(2%,
white solid).
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The compounds in the Table below were prepared by using an analogous method as
described for the preparation of compound 193, starting from the respective
starting
materials.
Compound number Structure Quantity Yield
mg 7%
Rsc:Compound 309 as a
mixture of 4
diasteoisomers (ratio:
5
65/35) N
(from intermediate 11
F3/
and 105)
5
EXAMPLE B36
Rs
F3C
PREPARATION OF COMPOUND 161
"R
÷÷c
F3C
COMPOUND 166:
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ccy
*s
cI31
F3C
AND COMPOUND 1197: -
In a sealed tube, under N2, 1,3-dimethy1-1H-pyrazole-4-carbaldehyde (284 mg;
2.28
mmol) and Ti(iPrO)4 (727 L; 3.05 mmol) were added to a solution of
intermediate 11
(500 mg; 1.52 mmol) in THF (10 mL). The solution was stirred at 50 C for 2h.
The
reaction mixture was cooled to 5 C and iPrMgC1 (3.8 mL; 7.61 mmol) was added
dropwise. The reaction mixture was allowed to rise slowly to rt and stirred
overnight.
The reaction mixture was poured onto a 10% aqueous solution of K2CO3 and
Et0Ac.
The insoluble was filtered through a pad of celite0 then, the organic layer
was decanted,
dried over MgSO4, filtered and the solvent was evaporated. The residue (866
mg, brown
oil) was purified by chromatography over silica gel (SiO2; 40 g; eluent: from
96% DCM,
4% Me0H, 0.4% NH4OH to 93% DCM, 7% Me0H, 0.7% NH4OH). The pure fractions
were collected and the solvent was evaporated to dryness. The residue (496 mg,
yellow
oil) was recrystallized with diethylether. The precipitate was filtered and
dried to give
324 mg of compound 161( (45%, white solid).
270 mg of compound 161( was purified by chiral SFC (CHIRALPAK AD-H Sum
250*30 mm, mobile phase: 70% CO2, 30% iPOH (0.3% iPrNH2)). The pure fractions
were collected and the solvent was evaporated to give 128 mg of compound 166
(18%,
colourless oil) and 131 mg of compound 167 (18%, colourless oil).
Compound 166 was freeze-dried with water-ACN to give 110 mg of compound 166
(15%, white solid).
Compound 167 was freeze-dried with water-ACN to give 115 mg of compound 167
(16%, white solid).
The compounds in the Table below were prepared by using an analogous method as
described for the preparation of compound 161, starting from the respective
starting
materials. The most relevant minor deviations are indicated in the column
Yield.
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Compound number Structure Quantity Yield
26 mg 4%
RS ---"N
Compound 172 (from
intermediate 11 and 1,5-
dimety1-1H-pyrazole-4-
carbaldehyde
C¨r)
/
F3c SN
109 mg 15%
"R N
Compound 173 and
compound 174 From
SFC purification of F3
compound
172CHIRALCEL OD-H Compound 173
gm 250*30 mm,
mobile phase: 80% CO2, cej
*s
20% Et0H (0.3%
iPrNH2)) N5
cnN 105 mg 14%
F3c SN
Compound 174
31 mg 9%
Compound 186 (from
*s
intermediate 70b and 1-
methyl-1H-Pyrazole-4-
/
carboxaldehyde (nN
F3C
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Compound number Structure Quantity Yield
N-)---- 53 mg 15%
.
*S C---\-/N---
C9N
N
Compound 188 and
compound 189 /
F3c s----,N--
From SFC purification
Compound 188
of compound 186(
CHIRALPAK AD-H 5
gm 250*30 mm, mobile c9:-.-_\_,N_____
*s 74 mg 21%
phase: 85% CO2, 15%
N
Et0H (0.3% iPrNH2)) N
F3C S----'",N".'
Compound 189
Compound 281 250 71%
Ti(OEt)4
From intermediate 70a .. ,7 71_ was used
and 1-methyl-1H- *R N in the
Pyrazole-4- N synthesis.
carboxaldehyde. /
F3c s-----,N"--.
77 mg 22%
Compound 190 and
N
compound 191
/
F3C S----Nr
From SFC purification
of compound Compound 190
281(CHIRALPAK AD-
H 5 gm 250*30 mm, )----- 61 mg 17%
mobile phase: 85% CO2,
( *R -',N/ -----
15% Et0H (0.3% N
iPrNH2))
Cril
/
F3c s----,N,
Compound 191
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EXAMPLE B38
F
F
RS
N
/ 1
/
N
S----
PREPARATION OF COMPOUND 170: F3C
Intermediate 1 lb (198 mg) and 1-(3,5-difluorophenyl)propan-2-one) (86 mg;
0.51 mmol)
in THF (5 mL). The mixture was stirred at rt overnight. Then NaBH(OAc)3 (161
mg;
0.76 mmol) was added portionwise. The mixture was stirred at rt for 24h. The
solution
was poured out into cooled water, basified with a solution of NaOH 3N, Et0Ac
was
added. The organic layer was separated, dried over MgSO4, filtered and
evaporated to
dryness. The residue (141 mg) was purified by chromatography over silica gel
(irregular
bare silica 40 g, mobile phase: 0.1% NH4OH, 97% DCM, 3% Me0H). The pure
fractions
were collected and the solvent was evaporated. The residue (50 mg) was freeze-
dried
with ACN/water 20/80 to give 16 mg of compound 170.
The compounds in the Table below were prepared by using an analogous method as
described for the preparation of compound 170, starting from the respective
starting
materials.
Compound number Structure Quantity Yield
35 mg
¨
RS
\ I
Compound 171 (from
intermediate 1 lb and 2- N5
methoxyphenylacetone
C¨rfõ,
/
F3C s-----N
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Compound number Structure Quantity Yield
F
38 mg
Compound 183 (from ciN RS
intermediate 1 lb (and 3-
fluorophenylacetone N)-
HCI salt
(fl
F3c/ S----N
55 mg
RS
Compound 182 (from c-3
intermediate lib and 3-
N)-
methyl-1-pheny12-
butanone e--)
/
F3c S-----N
0
/ \ 143 mg 56%
\./
Compound 185 (from N
intermediate 70a and
Tetrahydropyran-4- & *R
N
carbaldehyde [50675-
18-8) / I
/
F3c s---N-
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Compound number Structure Quantity Yield
124 mg 48%
Compound 187 (from c9N
intermediate 70b and
Tetrahydropyran-4-
carb aldehyde [50675-
18-8) / I '3
F3 C'
EXAMPLE B39
"R
HCI salt
/
F3C/
PREPARATION OF COMPOUND 184:
A mixture of intermediate 70a (250 mg; 0.73 mmol), isobutyraldehyde (200 L;
2.19
mmol) and AcOH (42 L; 0.73 mmol) in DCE (8 mL) was stirred at 50 C for 3h.
The
reaction mixture was cooled to rt and NaBH(OAc)3 (464 mg; 2.19 mmol) was
added.
The reaction mixture was stirred at rt overnight, poured onto a 10% aqueous
solution of
K2CO3 and extracted with DCM. The organic layer was decanted, dried over
MgSO4,
filtered and evaporated to dryness. The residue was purified by chromatography
over
silica gel (irregular SiOH, 24 g; mobile phase: gradient from 100% DCM to 10%
Me0H
(+10%NH4OH), 90% DCM). The pure fractions were collected and evaporated to
dryness. The hydrochloride salt was prepared: The residue (110 mg, 38%) was
dissolved
in ACN and HC1 4N in 1,4-dioxane (2eq.) was added. The solution was evaporated
to
dryness and taken up several times with ACN. The residue was crystallized from
Et20
yielding 120 mg of compound 184 (HC1 salt).
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The compound in the Table below were prepared by using an analogous method as
described for the preparation of compound 184, starting from the respective
starting
materials.
Compound number Structure Quantity Yield
35 mg 15%
Compound 288 (from
intermediate 70b and
isobutyraldehyde
(nN
F3c/ SN
EXAMPLE B40
I.
&N
/F eN1
PREPARATION OF COMPOUND 289 F
A solution of intermediate 70a (100 mg; 0.29 mmol), p-tolualdehyde (50 L;
0.35 mmol)
in dichloroethane (3 mL) was stirred at rt for 3h. NaBH(OAc)3 (124 mg; 0.58
mmol) was
added and the mixture was stirred at rt overnight. A 10% aqueous solution of
K2CO3 and
DCM were added. The organic layer was decanted, filtered through chromabond
and
evaporated to dryness. The residue was purified by chromatography over silica
gel
(irregular SiOH, 10g; mobile phase: gradient from 0% NH4OH, 0% Me0H, 100% DCM
to 0.5% NH4OH, 5% Me0H, 95% DCM). The pure fractions were collected and
evaporated to dryness. The residue was freeze dried from water/ACN (80/20; 10
mL)
yielding 73 mg (56%) of compound 289.
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The compounds in the Table below were prepared by using an analogous method as
described for the preparation of compound 289, starting from the respective
starting
materials. The most relevant minor deviations are indicated in the column
Yield.
Compound number Structure Quantity Yield
85 mg 65
Compound 290 (from
intermediate 70b and p-
IN
tolualdehyde) &N
F
S-----\N
F
F
89 mg 68%
I.
Compound 291 (from THF as
intermediate 70b and 4-
.N/Ni
solvent
fluorobenzaldehyde)
&N
/ 1 N
F
S---"N
F
F
F
86 mg 65%
I.
Compound 292 (from THF as
intermediate 70a and 4-
i/N1 solvent
fluorobenzaldehyde)
&N
/ 1 N
F
S---"N
F
F
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Compound number Structure Quantity Yield
68 mg 56%
Compound 293 (from / 'Zil'i
THF as
intermediate 70a and 3- CN
solvent
methylbutyraldehyde)
F , /
F/ F ( nN
S = - " - " \ N
72 mg 60%
7
Compound 294 (from
THF as
intermediate 70b and 3- (N
solvent
methylbutyraldehyde)
F , /
F/ F ( nl
S = - " - " \ N
\ 50 mg 39%
N-N
Q
Compound 295 (from N.-) THF as
intermediate 70a and 1- 1 T.Hi
methyl-1H-pyrazo le-4- CN solvent
carbaldehyde)
FAF I ,i
S---"\N%
F
0 50 mg 38%
Compound 296 (from
,:/N
intermediate 70a and N THF as
solvent
phenylacetaldehyde)
eir \ I
F .,./...
S N
F ----- \
F
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Compound number Structure Quantity Yield
lei 18 mg 11%
Compound 297 (from 1 s* iN 1 8 HCO2H
intermediate 70b and CN THF as
solvent
phenylacetaldehyde)
JN
F.... S N
/...
F -----\
7F
O Compound 298 (from F 20 mg 14%
intermediate 70a and 2- N
(4-
N
fluorophenyl)acetaldehy c----'LN
de) F/
1F
\ 74 mg 58%
N¨N
Compound 299 ((from S,
NJ THF as
intermediate 70b and 1-
methy1-1H-pyrazo le-4- (N solvent
carbaldehyde)
e------) NJ
F
F I 1
S---"\N%
F
EXAMPLE B41
H N R
N
1----b
N
F3C/
S-------
PREPARATION OF COMPOUND 273
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\¨
H NO ___________________ R.
*S
1---7-3NN
C)\1
S-'- F3C/
compound 200 NI
H NORS
L....7_3N
N
eY\I
F3C/
S'r\j-
and compound 222
A mixture of intermediate 71(546 mg; 1.01 mmol) and TFA (1.5 mL; 19.6 mmol) in
DCM (15 mL) was stirred at RT for 4 hours. The mixture was evaporated to
dryness.
The residue was taken up with DCM and H20 then, basified with aqueous NaOH 3N.
The organic layer was decanted, dried over MgSO4, filtered and evaporated to
dryness
yielding 400 mg (90%) of compound 222.
The enantiomers were separated by chiral SFC (Chiralpak AD-H 5 m 250*30mm;
mobile phase: 50% CO2, 50% Et0H (0.3% iPrNH2)). The pure fractions were
collected
and evaporated to dryness yielding 106 mg (24%) of compound 273 and 130 mg
(29%)
of compound 200.
Compound 273 can also be prepared from intermediate 71a using the same
procedure.
Compound 200 can also be prepared from intermediate 71b using the same
procedure.
The compounds in the table below were prepared using an analogous method as
described for the preparation of compound 273 starting from the respective
starting
materials.
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compound number Structure Quantity Yield
compound 211 125 mg 56%
(
(nN
N
From intermediate 77a
compound 213 123 mg 55%
F3C
From intermediate 77b
compound 259 96 mg 49%
Cr,j1
F3C
From intermediate 80
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compound number Structure Quantity Yield
H
compound 269 N 18 mg 55%
) ?F
ILlb
N
s---N
F3c/ From intermediate 82
H
compound 269a N 170 mg 99%
Q
) R* p( '
LbN
N
/ I
F3/ S-----\ N-
From intermediate 82a
H
compound 269b N 193 mg Quant.
) s?F
ILlb
N
/c fl)
/
F3c S-----N
From intermediate 82b
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EXAMPLE B42
N ___________________________________
/\NZ) *R EbN
(N11
F3C/
PREPARATION OF COMPOUND 274
\N ______________________ *S
N
(N11
F3C/
and compound 203
Under N2 at RT, 1-methyl-1H-pyrazole-4-carbaldehyde (100 mg; 0.913 mmol) and
Ti(OEt)4 (0.25 mL; 1.233 mmol) were added to a solution of intermediate 11(200
mg;
0.609 mmol) in THF (3 mL). The solution was stirred at room temperature for 20
hours.
The reaction mixture was cooled to 5 C and iPrMgC12M in THF (1.5 mL; 3.045
mmol)
was added dropwise. The reaction mixture was stirred for 30 min at 5 C,
allowed to rise
slowly RT over 6 hours and poured onto a cold aqueous solution of K2CO3. DCM
was
added and the reaction mixture was filtered through a pad of Celite0. The
insoluble
material was washed several times with DCM. The organic layer was decanted,
filtered
over chromabond and evaporated to dryness. The residue was purified by
chromatography over silica gel (irregular SiOH, 24g; mobile phase: gradient
from 0%
.. NH4OH, 0% Me0H, 100% DCM to 0.8% NH4OH, 8% Me0H, 92% DCM). The pure
fractions were collected and evaporated to dryness yielding 195 mg (69%) of
racemic
compound. The enantiomers were separated by chiral SFC (Lux Cellulose-2 5 m
250*30mm; mobile phase: 55% CO2, 45% Me0H (0.3% iPrNH2)). The two fractions
were freeze dried from water/ACN (80/20; 12 mL) yielding 80 mg (28%) of
compound
274 and 82 mg (29%) of compound 203.
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EXAMPLE B43
/NI
Nn
I NEb
F3C/ S
PREPARATION OF COMPOUND 197
/NI if NEb
F3C/ S
compound 198
Nn,,s;
F3C/ S
and compound 199
Under N2 at RT, 1-methyl-1H-pyrazole-4-carbaldehyde (99 mg; 0.9 mmol) and
Ti(OEt)4
(0.25 mL; 1.2 mmol) were added to a solution of intermediate 11 (197mg; 0.6
mmol) in
THF (3 mL), the solution was stirred at RT for 20 hours. The reaction mixture
was cooled
to 0 C and CH3MgBr (3M in Et20; 1 mL; 3 mmol) was added dropwise. The solution
was stirred for 30 min at 0 C and allowed to slowly rise RT for 6 hours. The
solution
was poured onto a mixture of cold water and aqueous saturated NH4C1 then Et0Ac
was
added. The mixture was filtered through a pad of celite and extracted with
Et0Ac. The
organic layer was dried over MgSO4, filtered and evaporated to dryness. The
residue was
purified by chromatography over silica gel (irregular SiOH, 24g; mobile phase:
gradient
from 0% NH4OH, 0% Me0H, 100% DCM to 0.5% NH4OH, 5% Me0H, 95% DCM).
The pure fractions were collected and evaporated to dryness yielding 130 mg
(50%) of
compound 197. The enantiomers were separated by chiral SFC (Lux Cellulose-4 5
m
250*21.2mm; mobile phase: 60% CO2, 40% Me0H (+0.3% iPrNH2)). The pure
fractions
were collected, evaporated and freeze dried from water/ACN (80/20) yielding 46
mg
(17%) of compound 198 and 45 mg (17%) of compound 199.
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The compounds in the table below were prepared using an analogous method as
described for the preparation of compound 274 starting from the respective
starting
materials.
Compound Structure Quantity Yield
number
compound 214 19 mg 13%
Nn s
N
K
1---bN
N
/ 1
F3C/ From intermediate 11, 1-Isopropyl- 1H-
pyrazo le-4- carb aldehyde and iPrMgC12M in
THF
compound 283 ND 143 mg 52%
I \ RS
N----- LbN
N
/ 1
/
F3C S--"N
compound 283
From intermediate 11, 1 -methyl-1H-
pyrazo le-4-carbaldehyde and EtMgBr 1M in
THF
ip- 58 mg 21%
compound 220 L.....b
N
F3C
/
S-----N-
and compound 220
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Compound Structure Quantity
Yield
number
0......0
N ---..
-s
compound 221
1----bNN 61 mg 22%
/ / I
F3C S
compound 221
From purification of compound 283 by Chiral
SFC: CHIRALPAK IC 5ium 250*21.2mm;
mobile phase: 60% CO2, 40% Et0H (0.3%
iPrNH2)
H
compound 223 N 28 mg 20%
R S
oN
0
N
e\I
F31 S----Nr
From intermediate 11, tert-butyl 4-formyl-
1H-pyrazo le-l-carbo xylate and MeMgBr 3M
in Et20
s
compound 228 . 77 mg 57%
/7
R S
oN
0
N
e\I
F31 S-----Nr
From intermediate 11, isothiazo le-4-
carbaldehyde and MeMgBr 3M in Et20
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Compound Structure Quantity
Yield
number
compound 230 343 mg 78%
NO RS
F3C/ SN
Compound 230
From intermediate 11, 1-methy1-1H-
pyrazo le-4-carb aldehyde and
isobutylmagnesium bromide 2M in Et20
compound 247 N
'R
L.7DN 118 mg 27%
F3C/ S
Compound 247
N---
and
114 mg 26%
compound 248
F3C/
Compound 248
From purificationof compound 230 by chiral
SFC: Lux Cellulose-2 5nm 250*30mm;
mobile phase: 55% CO2, 45% Me0H (0.3%
iPrNH2).
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Compound Structure Quantity Yield
number
0
compound 231 25 mg 19%
RS
(r)F3C S
From intermediate 11, isoxazo le-4-
carbaldehyde and MeMgBr 3M in Et20
compound 233 26 mg 19%
N
F3C SN
From intermediate 11, pyridine-3-
carboxaldehyde and MeMgBr 3M in Et20
compound 235 23 mg 8%
N
*R
LbN
e)2 F3C/
Compound 235
and
N--- 17 mg 6%
compound 236 ij*s
Cn\I
F3C/
Compound 236
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Compound Structure Quantity Yield
number
and N-D õ
. ---'s
compound 237 N /
27 mg 9%
, 4_,
()
F3c S"---N*
Compound 237
N--
I / *R
and N
compound 238 o
0
N
29 mg 10%
F3C S"----N
Compound 238
From intermediate 11, 1-methy1-1H-
pyrazole-4-carbaldehyde and sec-butyl
magnesium chloride 25%wt in THF
chiral SFC purification: CHIRALPAK AD-H
5gm 250*30mm; mobile phase: 80% CO2,
20% iPrOH (0.6% iPrNH2).
Compounds 237 and 238 were obtained after
an additional chiral SFC:
CHIRALPAK AD-H 5gm 250*30mm;
mobile phase: 80% CO2, 20% iPrOH (0.6%
iPrNH2) then chiral SFC: Chiralcel OD-H
5gm 250x21.2mm; mobile phase: 82% CO2,
18% iPrOH (0.3% iPrNH2).
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Compound Structure Quantity
Yield
number
compound 284 174 mg 50%
y-D RS
/IN
F3C/ SN
Compound 284
From intermediate 11, 1-methy1-1H-
pyrazole-4-carbaldehyde and n-butyl
magnesium chloride 2M in THF
58 mg 20%
compound 243
R
LbN
F3C/ SN
Compound 243
and
*S N 64 mg 22%
compound 244 o
(n\I
F3C
Compound 244
From chiral SFC separation of compound
384: CHIRALPAK IC 5ium 250*30mm;
mobile phase: 60% CO2, 40% Et0H (0.3%
iPrNH2).
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Compound Structure Quantity Yield
number
compound 285 N 75 mg 22%
H,---) lrtS
F3C/ SN
Compound 285
From intermediate 11, tert-butyl 4-formyl-
1H-pyrazo le-l-carboxylate and iPrMgC1 2M
in THF
compound 245 H N / 25 mg 7%
and F3 (nN
/ S
compound 245
N
compound 246 H *S
24 mg 7%
eni
F3/ SN
compound 246
From Chiral SFC separation of compound
285: CHIRALPAK AD-H 5ium 250*30mm;
mobile phase: 70% CO2, 30% Et0H (0.3%
iPrNH2)
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Compound Structure Quantity
Yield
number
compound 249
S N 25 mg 19%
¨/
s
1.....bN
N
Cn\I
F31 S----NI
From intermediate 11, thiazo le-5-
carbaldehyde and MeMgBr 3M in Et20
compound 286 / N 94 mg 71%
0 6s
LbN
N
C)1
F3C/ S----N
Compound 286
From intermediate 11, 1-methy1-1H-
imidazole-5-carbaldehyde [CAS39021-62-0]
and MeMgBr 3M in Et20
/
compound 253 N = 0
NI----N3 22 mg 16%
e--)F3c/ s-----N
and compound 253
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Compound Structure Quantity Yield
number
compound 254
22 mg 15%
e)\i
F3c/ S
compound 254
From Chiral SFC separation of compound
286: Chiralcel OD-H 5 m 250x21.2mm;
mobile phase: 70% CO2, 30% Me0H (0.3%
iPrNH2))
EXAMPLE B44
H2N
(N)
C = = N
_____________________________________ / I
F3C S
PREPARATION OF COMPOUND 195
Hydrazine monohydrate (344; 0.86 mmol) was added to a solution of intermediate
85
(100 mg; 0.17 mmol) in Et0H (4 mL). The solution was heated at 50 C for 2h30.
The
reaction mixture was poured into ice water and extracted with DCM. The organic
layer
was separated, dried over MgSO4, filtered and evaporated till dryness. The
residue was
purified by chromatography over silica gel (irregular SiOH, 12g; mobile phase:
90%
DCM, 10% Me0H (+10% NH4OH)). The pure fractions were collected and evaporated
to dryness. The residue was taken up with Et20 and evaporated to dryness
yielding 35
mg (45%) of compound 195.
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EXAMPLE B45
1
N
\ IN
H 0
RS
LbNN
C)\1
F3C/ S-----Nr
Preparation of compound 287 ,
I
N
17
/ :
H \ ) *_
R
1---7-3NN
C----)N
_______________________ / I
F3 C/
'
compound 201 N
1
H ND,,/Nini
-
1-bNN
e)\1
F3C/ ______________________ S ------'-
and compound 202 N----
A mixture of intermediate 87 (145 mg; 0.24 mmol) and TFA (0.7 mL; 9.15 mmol)
in
DCM (7 mL) was stirred at RT overnight. The reaction mixture was evaporated to
dryness. The residue was diluted with DCM and H20 then, basified with aqueous
NaOH
3N. The organic layer was decanted, dried over MgSO4, filtered and evaporated
to
dryness yielding 100 mg (83%) of compound 287.
The enantiomers were separated by chiral SFC (CHIRALPAK IC 5 m 250x20mm;
mobile phase: 50% CO2, 50% Me0H (+2% iPrNH2)). The fractions containing each
enantiomer were collected, evaporated to dryness and purified by reverse phase
chromatography (YMC-actus Triart-C18 10 m 30*150mm; mobile phase: gradient
from
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75% NH4HCO3 0.2%, 25% ACN to 35% NH4HCO3 0.2%, 65% ACN). The pure
fractions were collected, evaporated to dryness and freeze dried from
ACN/water (20/80)
yielding 21 mg (17%) of compound 201 and 23 mg (19%) of compound 202.
EXAMPLE B46
RS \
Lib
/ I
F3C/
PREPARATION OF COMPOUND 224
"R
______________________ / I
F3C
compound 251
N
*S
(N)
/ I j
S
and compound 252 F3C
Under N2, a mixture of intermediate 1 lb (1 g) and 5-acetyl-1-methylpyrazole
(168
mg1.35 mmol) in THF (15 mL) was stirred at rt overnight. Then, NaBH(OAc)3 (718
mg;
3.4 mmol) was added portion wise. The reaction mixture was stirred at room
temperature
for 72h, poured into cold water, basified with K2CO3 powder and extracted with
Et0Ac.
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The organic layer was dried over MgSO4, filtered and evaporated to dryness.
The residue
was purified by chromatography over silica gel (irregular SiOH, 12g; mobile
phase:
gradient from 100% DCM, 0% Me0H to 95% DCM, 5% Me0H, 0.3% NH4OH). The
fractions were collected and evaporated to dryness. The residue (300 mg) was
purified a
second time by reverse phase chromatography (YMC-actus Triart C18 10 m
30*150mm; mobile phase: gradient from 65% NH4HCO3 0.2% aq, 35% ACN to 25%
NH4HCO3 0.2% aq, 75% ACN). The pure fractions were collected, evaporated to
dryness
yielding 104 mg of compound 224.
Compound 224 was submitted to chiral SFC separation ((Stationary phase:
Chiralcel
OD-H 5 m 250x21.2mm, Mobile phase: 70% CO2, 30% Et0H(0.3% iPrNH2)).
The fractions containing the product were mixed, concentrated and freeze-dried
(ACN/water: 80/20) to afford 48 mg of compound 251 and 46 mg of compound 252.
The compounds in the table below were prepared using an analogous method as
described for the preparation of compound 224 starting from the respective
starting
materials.
compound Structure Quantity Yield
number
compound 232 I/ % 63 mg
RS \ ¨/
111---N-)
(\j
F3C/ S----N
From intermediate 1 lb and 4-acetylpyridine
compound 258 N \ 45 mg
> RS
N- N
1------b
N
e)\1
/
F3C SN
From intermediate 1 lb and 5-acetylpyrimidine
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EXAMPLE B47
rj1 RS 61
C)\1 HCI salt
PREPARATION OF COMPOUND 234 F3/ s
A mixture of intermediate 88 (130 mg; 0.225 mmol) and HC1 4M in 1,4-dioxane
(0.7
mL; 2.8 mmol) in Me0H (7 mL) was stirred at rt for 24 hours. The solution was
cooled
at 5 C and Et20 was added. The precipitate was filtered and dried yielding 111
mg of
compound 234 (HC1 salt).
EXAMPLE B48
HO
oN
N
F3C S
PREPARATION OF COMPOUND 270
OH
63N
/ I
F3C S
and compound 271
A mixture of intermediate 11(300 mg; 0.854 mmol) and (R)-Styrene oxide (293
L;
2.563 mmol) in Et0H (6 mL) was stirred at 60 C for 4 hours. The reaction
mixture was
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evaporated to dryness and the residue was purified by chromatography over
silica gel
(irregular SiOH, 12g; mobile phase: gradient from 0% NH4OH, 0% Me0H, 100% DCM
to 1% NH4OH, 10% Me0H, 90% DCM). The fractions containing the products were
collected and evaporated to dryness yielding 157 mg of an intermediate residue
which
was purified by reverse phase chromatography (YMC-actus Triart C18 10 m
30*150mm; mobile phase: gradient from 60% NH4HCO3 0.2% aq, 40% ACN to 40%
NH4HCO3 0.2% aq, 60% ACN). The fractions containing the products were
collected,
evaporated to dryness and freeze dried from water/ACN (80/20; 10 mL) yielding
58 mg
(15%) of compound 270 and 62 mg (16%) of compound 271.
EXAMPLE B50
0
s=== s
eni
F3C
PREPARATION OF COMPOUND 105
TFA (2.5 mL) was added to a solution of intermediate 93 (500 mg; 0.95 mmol) in
DCM
(25 mL) and the reaction mixture was stirred for 18 hours. The reaction
mixture was
poured onto a 10% aqueous solution of K2CO3 and extracted with DCM. The
organic
layer was separated, filtered over Chromabond and evaporated to dryness. The
residue
was purified by chromatography over silica gel (irregular SiOH, 10g; mobile
phase: 0.7%
NH4OH, 7% Me0H, 93% DCM). The pure fractions were collected and evaporated to
dryness. The residue was crystallized from diisopropyl ethyl ether and dried
yielding 120
mg (29%) of compound 105.
The compounds in the table below were prepared using an analogous method as
described for the preparation of compound 105 starting from the respective
starting
materials.
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compound or number Structure Quantity Yield
compound 110 0 78 mg 34%
0
/
F3C
From intermediate 96
compound 109 117 mg 41%
R EN1
Cni
F3C
From intermediate 98
compound 111 0 47 mg 15%
0 S.
iõ.=
/
F3C
From intermediate 99
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compound or number Structure Quantity Yield
compound 90 58 mg 71%
0 ..
Cni
F3C
From intermediate 100
EXAMPLE B51
0 .
="µ S
Cni
F3C
PREPARATION OF COMPOUND ov
HC14N in 1,4-dioxane (0.708 mL; 2.833 mmol) was added to a solution of
intermediate
.. 101 (150 mg; 0.283 mmol) in ACN (7.5 mL) and the reaction mixture was
stirred at room
temperature overnight. The reaction mixture was poured onto a 10% aqueous
solution of
K2CO3 and extracted with DCM. The organic layer was decanted, filtered over
chromabond and evaporated to dryness. The residue was purified by
chromatography
over silica gel (irregular SiOH, 12g; mobile phase: gradient from 0% Me0H,
100% DCM
.. to 15% Me0H, 85% DCM). The pure fractions were collected and evaporated to
dryness.
The residue was taken up several times with Et20 yielding 30 mg (25%) of
compound
89.
The compounds in the table below were prepared using an analogous method as
described for the preparation of compound 89 starting from the respective
starting
.. materials.
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compound number Structure Quantity Yield
compound 87 47 mg 29%
1,== s ,F1
(T)\1
F3I
From intermediate 102
compound 86 55 mg 42%
N N
/ I jF3C/
From intermediate 102
Compound 311 181 mg 58%
HN
/ I
From intermediate 107
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EXAMPLE B56
RS . F
HCI salt
(N)
(¨)\'
, s,NV
F3C,
PREPARATION OF COMPOUND 317:
Intermediate 11(266 mg, 0.812 mmol), intermediate 110 (377 mg, 1.62 mmol) and
K2CO3 in ACN (8 mL) were stirred overnight at 90 C. The mixture was poured
into
water then extracted with Et0Ac. The organic layer was dried over MgSO4,
filtered and
evaporated to dryness. The residue was purified by chromatography over silica
gel
(Stationary phase: irregular SiOH 15-40gm 40g, mobile phase: DCM/MeOH:
gradient
from 100/0 to 92/8). The residue was purified by chromatography over silica
gel by
reverse phase (stationary phase: YMC-actus Triart-C18 10 gm 30*150mm, mobile
phase:
0.2% NH4HCO3/ACN: gradient from 60/40 to 0/100). The fractions containing the
product were collected and evaporated to dryness. The resulting residue was
solubilized
in ACN and 2 equivalents of a 4N solution of HC1 in dioxane was added. The
mixture
was concentrated and, then freeze-dried with acetonitrile/water 20/80 yielding
0.091 g of
compound 317 (HC1 salt).
EXAMPLE B57
0¨
L11)
N
TFA salt
N
/ I )
F S Nr
n F F
PREPARATION OF COMPOUND 320
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To a solution of intermediate 1 lb (200 mg) in ethanol (5 mL) was added 1- (3-
methoxyphenyl)propan-2-one (200 mg , 1.218 mmol), Pt02 (20 mg) and AcOH (2
drops). After stirring at 60 C overnight under H2, the reaction mixture was
concentrated
to give a residue which was purified by prep-HPLC (Column: SunFire 19*250mm
10um,
Mobile Phase A: 0.1%TFA/H20, B: ACN) to give 35 mg (12%) of compound 320 as
yellow solid.
The compounds in the table below were prepared using an analogous method as
described for the preparation of compound 320 starting from the respective
starting
materials.
compound Structure Quantity Yield
number
compound 321 / 44 mg
0
RS
N
Lb
N
/
F 1 1\1
S----\N)
F
F
TFA salt
From intermediate 1 lb and 1- (4-
methoxyphenyl)propan-2-one
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compound Structure Quantity Yield
number
Compound 322
11 F 54 mg
RS
N
FAF Cj\I
Si\I
F
TFA salt
From intermediate 1 lb and 2-
Fluorophenylacetone
Compound 323 HO 36 mg
I/
RS
NLb
N
FAF Cn\I
S------N
F
TFA salt
From intermediate 1 lb and 4-
hydroxyphenylacetone
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compound Structure Quantity Yield
number
Compound 324 40 mg
I/
RS
N
FAF Cj\I
Si\I
F
TFA salt
From intermediate 1 lb and 1- (p-
tolyl)propan-2-one
EXAMPLE B57
COMPOUND 61 was synthesized together with intermediate 68, 68a and 68b. See
synthesis
protocol for intermediate 68, 68a and 68b.
CONVERSION
CONVERSION Cl
*R
v-3
&N-
(n\I
F3C/ S----
Preparation of compound 196 N
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A solution of compound 273 (106 mg; 0.24 mmol), aqueous formaldehyde 37% w/w
(110 L; 1.48 mmol) and MgSO4 (580 mg; 4.83 mmol) in DCM (5 mL) was stirred at
RT for 1 hour. NaBH(OAc)3 (614 mg; 2.9 mmol) was added and the reaction
mixture
was stirred at RT for 15 hours. The solution was poured into iced water,
basified with
K2CO3 and extracted with DCM (x2). The organic layer was washed with brine,
dried
over MgSO4, filtered and evaporated to dryness. The residue was purified by
chromatography over silica gel (irregular SiOH, 10g, mobile phase: gradient
from 100%
DCM, 0% Me0H, 0% NH4OH to 90% DCM, 10% Me0H, 0.5% NH4OH). The pure
fractions were collected, evaporated to dryness and the residue was freeze
dried from
ACN/water (20/80) yielding 70 mg (64%) of compound 196.
The compounds in the table below were prepared using an analogous method as
described for the preparation of compound 196 starting from the respective
starting
materials.
compound number Structure Quantity Yield
compound 304a 50 mg 28%
(n2 F3/ SN
From compound 269a
Compound 304b 52 m 30%
(5-\
/ I j
F3C
From compound 269b
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CONVERSION C2
\¨
N _____________________________________
*S \ ¨ .1
/ I
S-----\
F3c/
PREPARATION OF COMPOUND 216
A solution of compound 200 (110 mg; 0.25 mmol) and aqueous formaldehyde 37%
w/w
(19 L; 0.25 mmol) in Me0H (5 mL) was stirred at RT for 3 hours. NaBH4 (19 mg;
0.5
mmol) was added and the reaction mixture was stirred at RT for 15 hours,
poured into
ice water, basified with K2CO3 and extracted with DCM (x2). The organic layer
was
washed with brine then dried over MgSO4, filtered and evaporated to dryness.
The
residue was purified by chromatography over silica gel (irregular SiOH 300g;
mobile
phase: gradient from 0.1% NH4OH, 5% Me0H, 95% DCM to 1% NH4OH, 10% Me0H,
90% DCM). The pure fractions were collected, evaporated to dryness and freeze-
dried
from ACN/water 20/80 yielding 15 mg (13%) of compound 216.
CONVERSION C3
N
1------b
N
F
SN
F
F
PREPARATION OF COMPOUND 303
Under N2 at 10 C, ethyl bromide (454; 0.6mmo1) was added to a solution of
compound
222 (150mg; 0.34mm01) and DIPEA (207 L; 1.2mm01) in THF (3mL). The solution
was
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stirred at rt overnight, then, poured onto cooled water. The product was
extracted with
Et0Ac. The organic layer was dried over MgSO4, filtered and evaporated to
dryness.
The residue (164mg) was purified by silica gel chromatography (Stationary
phase:
irregular bare silica 12g, Mobile phase: Gradient from 100% DCM, 0% Me0H (+10%
NH4OH) to 90% DCM, 10% Me0H (+10% NH4OH)). The fractions containing the
product were mixed and concentrated to afford 77 mg of an intermediate
fraction which
was further purified by via reverse phase (Stationary phase: YMC-actus Triart
C18 10 m
30*150mm, Mobile phase: Gradient from 65% NH4HCO3 0.2% , 35% ACN to 25%
NH4HCO3 0.2% , 75% ACN). The fractions containing the product were mixed and
concentrated to afford 40mg of a residue which was freeze-dried with
acetonitrile/water
20/80 to give 34mg (21%) of compound 303 as a white powder.
CONVERSION C4
( \I¨
) _______________________________________ (S
Ntb
N
F
S-----N
F
, , F
PREPARATION OF COMPOUND 31 o
A solution of compound 315 (114 mg; 0.237 mmol), formaldehyde, 37% in water
(106
L; 1.42 mmol) and MgSO4 (568 mg) in DCM (5 mL) was stirred at rt for 1 hour.
Then,
NaBH(OAc)3 (602 mg; 2.84 mmol) was added and the mixture was stirred at rt
overnight.
The reaction mixture was diluted with DCM and basified with a 10% aqueous
solution
of K2CO3. The organic layer was decanted, dried over MgSO4, filtered and
evaporated
to dryness. The residue was purified by chromatography over silica gel
(irregular SiOH,
12g; mobile phase: gradient from 0% NH4OH, 0% Me0H, 100% DCM to 1% NH4OH,
10% Me0H, 90% DCM). The fractions containing the product were collected,
evaporated to dryness and freeze dried from water/ACN (80/20; 10 mL) yielding
110 mg
(94%) of compound 316.
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ANALYTICAL PART
LCMS (LIQUID CHROMATOGRAPHY/MASS SPECTROMETRY)
General procedure
The High Performance Liquid Chromatography (HPLC) measurement was performed
using a LC pump, a diode-array (DAD) or a UV detector and a column as
specified in
the respective methods. If necessary, additional detectors were included (see
table of
methods below).
Flow from the column was brought to the Mass Spectrometer (MS) which was
configured with an atmospheric pressure ion source. It is within the knowledge
of the
skilled person to set the tune parameters (e.g. scanning range, dwell time...)
in order to
obtain ions allowing the identification of the compound's nominal monoisotopic
molecular weight (MW). Data acquisition was performed with appropriate
software.
Compounds are described by their experimental retention times (Rt) and ions.
If not
specified differently in the table of data, the reported molecular ion
corresponds to the
[M+H]+ (protonated molecule) and/or EM-Ht (deprotonated molecule). In case the
compound was not directly ionizable the type of adduct is specified (i.e.
[M+NH4]
[M+HCOO], etc...). For molecules with multiple isotopic patterns (Br, Cl..),
the
reported value is the one obtained for the lowest isotope mass. All results
were obtained
with experimental uncertainties that are commonly associated with the method
used.
Hereinafter, "SQD" means Single Quadrupole Detector, "RT" room temperature,
"BEH"
bridged ethylsiloxane/silica hybrid, "HSS" High Strength Silica, "DAD" Diode
Array
Detector. All other abbreviations used are as defined before.
Table la: LCMS Method codes (Flow expressed in mL/min; column temperature (T)
in
C; Run time in minutes).
Flow
Run
Method Mobile
Instrument Column Gradient tim
code phase Column
A: CI-3COOH 90% A for 0.8min,
Agilent: Phenomene
0.1% i = 0 8
in water, to 20% A in =
1200 - x: Luna-
1 B: CI-3COOH 3.7min, held for ---- 10
DAD and C18 (5 m,
0.05% in 3min, back to
MSD6110 2 x5Omm) 50
CH3CN 90% A in 2min.
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Flow
Run
Method Mobile
Instrument Column Gradient tim
code phase Column
A: CI-3COOH 100% A for lmin,
Agilent: Phenomene 0 8
=
0.1% in water, to 40% A in 4mm, =
1200 - x: Luna-
2 B: CI-3COOH to 15% A in ---- 10
DAD and C18 (5 m,
0.05% in 2.5min, back to
MSD6110 2 x5Omm) 50
CH3CN 100% A in 2min.
84.2% A for
Waters:
Waters: A: 95% 0.49min, to 10.5%
Acquity
BEH C18 CH3COONH4 A in 2.18min, held 0'343
UPLC -
3 (1.7 m, 7m1M / 5% for 1.94min, back ---- 6.2
DAD and
2.1x100m CH3CN, B: to 84.2% A in
Quattro 40
m) CH3CN 0.73min, held for
Micro TM
0.73min.
84.2% A to 10.5%
Waters: Waters: A: 95%
A in 2.18 min, 0.343
Acquity BEH C18 CH3COONH4
4 held for 1.96 min
H-Class - (1.7 m, 7m1M / 5% 6.1
back to 84.2% A
DAD and 2.1x100m CH3CN, B: 40
in 0.73 min, held
SQD2TM CH3CN
for 0.73 min.
Shimadzu: SunFire A: HCOOH 70% A for 2.0 2.6
LC- C18 5i,tm 0.1%inwater, 0.4min, to 5% A
MS2020 - 50*4.6mm B:HCOOH in 1.2 min, to 1 %
SPD- 0.1%in A in 1.0 min. 40
M20A and CH3CN
Alltech
3300ELS
6 Waters ACQUIT A: HCOOH 90% A for 0.6 2.0
UPLC- Y UPLC 0.1%inwater, 0.1min, to 5% A
QDa- BEH C18 B:HCOOH in 1.1 min, hold 5
PDA 1.7gm 0.1%in % A in 0.8 min. 50
Detector 2.1*50mm CH3CN
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Flow
Run
Method Mobile ----
Instrument Column Gradient tim
code phase Column
e
T
7 Waters ACQUITY A:HCOOH 80% A for 1.0 0.4 9.0
UPLC- UPLC BEH 0.1%inwater, min, to 5% A in
QDa- C18 1.7[un B:HCOOH 7.0 min, hold 5 %
PDA 2.1*100mm 0.1%in A in 1.0 min. 50
Detector CH3CN
Table lb: LCMS data. Co. No. means compound number; Rt means retention time in
min.
Co. Adduct/ LCMS
Rt (min) [M+H]+
No. [M-11]- Method
1 2.725 419.2 - 1
2 2.88 425.1 483.4 [M+CH3COO] 3
3 2.9 447.2 - 1
4 2.26 421.1 479.4 [M+CH3COO] 3
3.314 459 2
6 3.176 459 - 2
7 3.129 459 - 2
8 3.71 454.9 - 2
9 3.333 421 2
3.269 421 - 2
11 3.652 454.9 - 2
12 3.409 421 2
13 2.498 385 - 1
14 2.76 405 463.4 [M+CH3COO] 3
3.516 455 2
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Co. Adduct/ LCMS
Rt (min) [M+H]+
No. [M-11]- Method
16 2.88 419.5 463.3 [M+CH3COO] 4
17 3.074 482.9 - 1
18 2.94 447 - 1
19 3.92 461.2 519.7 [M+CH3COO] 3
20a 3.15 433.1 491.4 [M+CH3COO] 3
20b 3.15 433.1 491.4 [M+CH3COO] 3
21a 3.66 447.4 445.4 3
21b 3.65 447.3 445.3 3
22 3.104 437 - 1
23 2.11 423.5 481.3 [M+CH3COO] 4
24 2.46 452.5 510.3 [M+CH3COO] 4
25 2.43 - 466.3 [M+CH3COO] 3
26 3.28 420.1 - 3
28 2.08 409.1 - 3
29 2.09 409.1 - 3
30 2.17 423.2 481.4 [M+CH3COO] 3
31 2.67 461 519.3 [M+CH3COO] 3
34 2.16 409.1 467.4 [M+CH3COO] 3
35 2.26 423.2 481.4 [M+CH3COO] 3
36 2.39 451.2 509.5 [M+CH3COO] 3
37 2.375 492 - 1
40 1.98 395.1 - 3
41 1.95 395.4 - 4
44 2.31 371.5 429.2 [M+CH3COO] 4
45 2.936 433 - 1
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Co. Adduct/ LCMS
Rt (min) [M+H]+
No. [M-11]- Method
46 2.871 433 - 1
47 3.05 463 - 1
48 2.79 545.9 - 1
49 2.901 433 - 1
50 3.019 447 - 1
54 2.66 436.9 - 1
56 2.731 436.9 - 1
57 3.604 449 - 2
58 2.67 385 443.2 [M+CH3C00]- 3
59 2.2 423 481.2 [M+CH3C00]- 3
61 2.11 357 415.1 [M+CH3C00]- 3
62 2.45 495.2 - 3
63 2.55 455.2 513.4 [M+CH3C00]- 3
64 2.92 411 469.1 [M+CH3C00]- 3
65 2.46 413.1 471.2 [M+CH3C00]- 3
66 2.31 427.1 485.2 [M+CH3C00]- 3
67 2.25 441.5 499.4 [M+CH3C00]- 4
69 2.89 433.1 491.3 [M+CH3C00]- 3
71 2.75 509.2 - 3
74 2.54 547.4 605.7 [M+CH3C00]- 3
75 2.56 506.3 564.5 [M+CH3C00]- 3
76 2.23 462.2 520.4 [M+CH3C00]- 3
77 2.01 452.1 520.5 [M+CH3C00]- 3
82 1.95 426.4 484.2 [M+CH3C00]- 4
84 3.05 465.1 523.3 [M+CH3C00]- 3
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Co. Adduct/ LCMS
Rt (min) [M+H]+
No. [M-11]- Method
88 2.13 488.4 546.3 [M+CH3COO] 3
112 2.24 452.1 510.4 [M+CH3COO] 3
86 2.20 452.2 510.5[M+CH3C00-] 3
87 2.13 438.2 496.6[M+CH3C00-] 3
89 2.14 430.1 488.3[M+CH3C00-] 3
90 2.32 448.1 506.4[M+CH3C00-] 3
105 1.96 426.4 484.3[M+CH3C00-] 4
108 2.34 452.2 510.4[M+CH3C00-] 3
109 2.13 438.1 496.4[M+CH3C00-] 3
110 1.93 442.5 500.2[M+CH3C00-] 4
111 1.90 442.5 500.3[M+CH3C00-] 4
163 2.42 489.1 547.4[M+CH3C00-] 3
164 2.43 489.2 547.4[M+CH3C00-] 3
195 1.90 452.5 510.3[M+CH3C00-] 4
196 2.35 454.2 512.4[M+CH3C00-] 3
197 2.29 437.2 495.5[M+CH3C00-] 3
198 2.25 437.2 495.4[M+CH3C00-] 3
199 2.25 437.2 495.5[M+CH3C00-] 3
200 2.28 440.1 498.3[M+CH3C00-] 3
201 1.92 506.5 564.4[M+CH3C00-] 4
202 1.89 506.5 564.5[M+CH3C00-] 4
203 2.70 465.2 523.4[M+CH3C00-] 3
211 2.40 454.1 512.3[M+CH3C00-] 3
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Co. Adduct/ LCMS
Rt (min) [M+H]+
No. [M-11]- Method
213 2.38 454.1 512.3[M+CH3C00-] 3
214 2.98 493.3 551.5[M+CH3C00-] 3
220 2.43 451.1 509.3[M+CH3C00-] 3
221 2.44 451.1 509.3[M+CH3C00-] 3
222 2.31 440.1 498.3[M+CH3C00-] 3
223 2.20 423.0 421.1 3
224 2.51 437.1 495.3[M+CH3C00-] 3
228 2.68 440.0 498.2[M+CH3C00-] 3
230 2.70 479.2 537.4[M+CH3C00-] 3
231 2.59 424.1 482.3[M+CH3C00-] 3
232 2.58 434.1 492.3[M+CH3C00-] 3
233 2.56 434.1 492.3[M+CH3C00-] 3
234 2.03 478.2 536.4[M+CH3C00-] 3
235 2.91 479.2 537.4[M+CH3C00-] 3
236 2.91 479.2 537.4[M+CH3C00-] 3
237 2.89 479.3 537.5[M+CH3C00-] 3
238 2.89 479.2 537.5[M+CH3C00-] 3
243 2.75 479.2 537.4[M+CH3C00-] 3
244 2.75 479.2 537.4[M+CH3C00-] 3
245 2.54 451.1 449.2 3
246 2.53 451.1 449.2 3
247 2.69 479.2 537.4[M+CH3C00-] 3
248 2.69 479.2 537.4[M+CH3C00-] 3
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Co. Adduct/ LCMS
Rt (min) [M+H]+
No. [M-11]- Method
249 2.59 440.0 498.2[M+CH3C00-] 3
250 2.42 489.1 547.3 [M+CH3C00-] 3
251 2.50 437.1 495.3 [M+CH3C00-] 3
252 2.50 437.1 495.3 [M+CH3C00-] 3
253 2.34 437.1 495.4[M+CH3C00-] 3
254 2.34 437.1 495.3 [M+CH3C00-] 3
258 2.42 435.1 493.2[M+CH3C00-] 3
259 2.16 426.1 484.3 [M+CH3C00-] 3
269 2.34 458.4 516.4[M+CH3C00-] 4
270 2.52 449.4 507.4[M+CH3C00-] 4
271 2.48 449.4 507.4[M+CH3C00-] 4
273 2.29 440.2 498.5 [M+CH3C00-] 3
274 2.71 465.2 523.4[M+CH3C00-] 3
133 2.36 463.2 521.5 [M+CH3C00]- 3
137 2.26 463.5 521.4 [M+CH3C00]- 4
138 2.27 463.5 521.4 [M+CH3C00]- 4
145 2.42 441.1 499.4 [M+CH3C00]- 3
147 2.11 440.1 498.2 [M+CH3C00]- 3
146 2.10 440.1 498.2 [M+CH3C00]- 3
148 2.35 468.2 466.4 3
154 2.46 441.1 499.4 [M+CH3C00]- 3
155 2.44 441.1 499.3 [M+CH3C00]- 3
470.2 [M+CH3C00]-;
150 2.05 412.0 3
410.0
527.5 [M+CH3C00]-;
193 2.96 469.2 3
467.8
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Co. Adduct/ LCMS
Rt (min) [M+H]+
No. [M-11]- Method
527.5 [M+CH3C00]-;
162 2.96 469.2 3
467.8
527.4 [M+CH3C00]-;
156 2.96 469.2 3
467.2
527.4 [M+CH3C00]-;
157 2.81 469.2 3
467.2
158 2.60 427.1 485.3 [M+CH3C00]- 3
159 2.60 427.1 485.3 [M+CH3C00]- 3
537.4 [M+CH3C00]-;
161 2.76 479.2 3
477.2
537.3 [M+CH3C00]-;
166 2.76 479.2 3
477.2
537.3 [M+CH3C00]-;
167 2.75 479.2 3
477.1
537.4 [M+CH3C00]-;
172 2.75 479.2 3
477.4
537.4 [M+CH3C00]-;
173 2.76 479.2 3
477.3
537.4 [M+CH3C00]-;
174 2.75 479.2 3
477.2
526.4 [M+CH3C00]-;
168 2.40 468.2 3
466.3
526.4 [M+CH3C00]-;
169 2.39 468.2 3
466.3
170 3.20 483.1 541.3 [M+CH3C00]- 3
171 2.93 477.2 535.4 [M+CH3C00]- 3
183 3.09 465.1 523.4 [M+CH3C00]- 3
182 3.48 475.2 533.4 [M+CH3C00]- 3
192 2.90 483.5 541.4 [M+CH3C00]- 4
179 3.03 483.2 541.5 [M+CH3C00]- 3
180 3.04 483.3 541.5 [M+CH3C00]- 3
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Co. Adduct/ LCMS
Rt (min) [M+H]+
No. [M-11]- Method
184 2.73 399.4 457.3 [M+CH3COOt 4
457.2 [M+CH3C00]-;
288 2.74 399.1 3
397.1
185 2.48 441.5 499.4 [M+CH3COO] 4
499.3 [M+CH3C00]-;
187 2.42 441.1 3
439.4
537.5 186 2.87 479.2 [M+CH3C00]-;3
477.2
537.4 [M+CH3C00]-;
188 2.80 479.5 4
523.4 [M+HCOO]
537.4 1 [M+CH3C00]-;
89 2.78 479.5 4
523.2 [M+HCOO]
190 2.80 479.5 537.4 [M+CH3COO] 4
191 2.79 479.5 537.4 [M+CH3COO] 4
289 3.32 447 505 [M+CH3COO] 3
290 3.33 447 505 [M+CH3COO] 3
291 3.25 451 509 [M+CH3COO] 3
292 3.25 451 509 [M+CH3COO] 3
293 2.78 413 471 [M+CH3COO] 3
294 2.77 413 471 [M+CH3COO] 3
295 2.24 437 495 [M+CH3COO] 3
296 3.07 447 505 [M+CH3COO] 3
297 3.07 447 505 [M+CH3COO] 3
298 3.11 465 523 [M+CH3COO] 3
299 2.25 437 495 [M+CH3COO] 3
303 2.42 468 526 [M+CH3COO] 3
304a 2.69 472 530 [M+CH3COO] 3
304b 2.70 472 530 [M+CH3COO] 3
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Co. Adduct/ LCMS
Rt (min) [M+H]+
No. [M-11]- Method
307 3.51 493 551 [M+CH3COO]- 3
2.92 &
309 455 513 [M+CH3COO]- 3
3.02
311 1.99 426 484 [M+CH3COO]- 3
312 3.76 537 595 [M+CH3COO]- 3
313 3.76 537 595 [M+CH3COO]- 3
314 2.90 472 530 [M+CH3COO]- 3
315 2.22 482 540 [M+CH3COO]- 3
316 2.30 496 554 [M+CH3COO]- 3
317 3.14 465 - 3
318 2.57 502 - 3
320 0.893 477.2 - 5
321 1.300 477.4 - 6
322 6.213 465.3 - 7
323 1.203 463.3 - 6
324 0.692 461.2 - 5
SFCMS-METHODS
General procedure for SFC-MS methods
The SFC measurement was performed using an Analytical Supercritical fluid
chromatography (SFC) system composed by a binary pump for delivering carbon
dioxide
(CO2) and modifier, an autosampler, a column oven, a diode array detector
equipped with
a high-pressure flow cell standing up to 400 bars. If configured with a Mass
Spectrometer
(MS) the flow from the column was brought to the (MS). It is within the
knowledge of
the skilled person to set the tune parameters (e.g. scanning range, dwell
time...) in order
to obtain ions allowing the identification of the compound's nominal
monoisotopic
molecular weight (MW). Data acquisition was performed with appropriate
software.
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Table 2a. Analytical SFC-MS Methods (Flow expressed in mL/min; column
temperature
(T) in C; Run time in minutes; Backpressure (BPR) in bars; all other
abbreviations used
in the table below are as defined before)
Flow Run time
Method code column mobile phase gradient
Col T BPR
1 Phenomenex 3
A: CO2 3.5
Luxcellulose-4 30% B
B: Me0H
column (3 [tm, hold 3 min' (0.3% iPrNH2) 35
100 x 4.6 mm) 103
2 Phenomenex 3
A: CO2 3.5
Luxcellulose-2 25% B
B: Me0H
column (3 [tm, hold 3 min' (0.3% iPrNH2) 35
100 x 4.6 mm) 103
Daicel A: CO2
3
3 Chiralpak0 B:
30%B 3.5
Et0H/iPrOH
hold 3min' 35
AD-3 (3 lina, 50/50 (0.3% 103
100 x 4.6 mm) iPrNH2)
Daicel 3
4 A: CO2 3.5
Chiralpak0 25%B
B: Et0H (0.3%
AD-3 (3 [tm, hold 3 min' 35 iPrNH2)
100 x 4.6 mm) 103
Daicel 3
A: CO2 3.5
Chiralpak0 30%B
B: Et0H (0.3%
AD-3 (3 [tm, hold 3 min' 35 iPrNH2)
100 x 4.6 mm) 103
Daicel A: CO2 4
7 3.5
Chiralpak AD- B: Et0H 50% B
3(3 [tm, 100 x hold 4 min' 35
4.6 mm) (0.3% iPrNH2) 103
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Flow Run time
Method code column mobile phase gradient
Col T BPR
Daicel A: CO2 6
8 3.5
Chiralpak AD- B: iPrOH 20% B
3 (3 [tm, 100 x hold 6min' 35
4.6 mm) (0.3% iPrNH2) 103
Daicel 3
9 A: CO2 3.5
Chiralpak AD- 30% B
B: iPrOH
3 (3 [tm, 100 x hold 3 min' 35 (0.3% iPrNH2)
4.6 mm) 103
Daicel 3
3.5
Chiralpale A: CO2 AD- 40%B
B: iPrOH
3 (3 [tm, 100 x hold 3 min' 35 (0.3% iPrNH2)
4.6 mm) 103
Daicel 3
11 A: CO2 3.5
Chiralpak0 25%B
B: Me0H
AD-3 (3 [tm, hold 3 min' 35 (0.3% iPrNH2)
100 x 4.6 mm) 103
12 A: CO2
Daicel 3
3.5
Chiralpak0 B: Me0H 30% B
AD-3 (3 [tm, (+0.3% hold 3 min' 35 103
100 x 4.6 mm) iPrNH2)
Phenomenex
A: CO2 6
16 3.5
Lux cellulose 2 B: Me0H 40% B
hold 6 min' 35
(3 [tm, 100 x
(0.3% iPrNH2) 103
4.6 mm)
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Flow Run time
Method code column mobile phase gradient
Col T BPR
Phenomenex
A: CO2 10
50%B 3.5
17 Lux cellulose 2 B: Me0H hold 10
(3 1..tm, 100 x 35
(0.3% iPrNH2) mill, 103
4.6 mm)
Daicel 3
18 A: CO2 3.5
Chiralpak0 IC- 40% B
B: Et0H (0.3%
3 (3 i.tm, 100 x hold 3 min,
iPrNH2) 35
4.6 mm) 103
Daicel 3
20 A: CO2 3.5
Chiralce10 OD- 20% B
B: Et0H (0.3%
3 (3 iAm, 100 x hold 3 mill,iPrNH2) 35
4.6 mm) 103
21 Daicel 3
A: CO2 3.5
Chiralce10 OD- 30% B
B: Et0H (0.3%
3 (3 i_tm, 100 x hold 3 min,
iPrNH2) 35
4.6 mm) 103
22 Daicel 3
A: CO2 3.5
Chiralce10 OD- 20% B
B: iPrOH
3 (3 1..tm, 100 x hold 3 min,
(0.3% iPrNH2) 35
4.6 mm 103
23 Daicel 3
A: CO2 3.5
Chiralce10 OD- 30% B
B: Me0H
3 (3 1..tm, 100 x hold 3 min,
(0.3% iPrNH2) 35
4.6 mm 103
24 Daicel 3
A: CO2 3.5
Chiralce10 OJ- 15% B
B: Et0H (0.3%
3 (3 i_tm, 100 x hold 3 min,
iPrNH2) 35
4.6 mm 103
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Flow Run time
Method code column mobile phase gradient
Col T BPR
25 Daicel 3
A: CO2 3.5
Chiralce10 OJ- 10% B
B: Me0H
3 (3 [Lm, 100 x hold 3 min' 35 (0.3% iPrNH2)
4.6 mm 103
26 Daicel 3
A: CO2 3.5
Chiralce10 OJ- 30% B
B: Me0H
3 (3 [Lm, 100 x hold 3 min' 35 (0.3% iPrNH2)
4.6 mm 103
Daicel 3
37 A: CO2 3.5
Chiralpak0 IC- 40% B
B: iPrOH
3 (3 [tm, 100 x hold 3 min' 35 (0.3% iPrNH2)
4.6 mm) 103
Daicel 6
38 A: CO2 3.5
Chiralpak0 IC- 20% B
B: Me0H
3 (3 [tm, 100 x hold 6 min' 35 (0.3% iPrNH2)
4.6 mm) 103
39 Daicel 3
A: CO2 3.5
Chiralce10 OD- 15% B
B: Me0H
3 (3 [tm, 100 x hold 3 min' 35 (0.3% iPrNH2)
4.6 mm 103
Phenomenex
A: CO2
40 3.5 3
40%B
Lux cellulose 2
B: Et0H hold 3 min'
(3 [tm, 100 x
35 103
(0.3% iPrNH2)
4.6 mm)
Table 2b. SFC-MS data. (Isomer elution order 'A' elutes before 'IT under the
described
conditions)
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Isomer
Co. Rt SFCMS
UV% Area elution
No. (min) Method
order
20a 2.62 100 A 1
20b 3.00 99.71 B 1
21a 2.66 98.09 A 2
21b 3.01 96.93 B 2
82 1.49 99.72 A 5
105 1.65 97.6 B 5
110 1.32 97.6 A 12
111 2.27 100 B 12
163 0.94 100 A 26
164 1.44 100 B 26
198 1.54 100 A 1
199 2.29 100 B 1
273 0.75 99.6 A 7
200 1.19 99.6 B 7
274 1.29 100 A 2
203 2.04 100 B 2
211 1.58 100 A 12
213 2.22 99.05 B 12
220 1.29 100 A 18
221 1.90 100 B 18
235* 2.81 100 A 8
236* 3.46 100 B 8
237* 1.97 100 A 22
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Isomer
Co. Rt SFCMS
UV% Area elution
No. (min) Method
order
238* 2.43 100 B 22
243 1.26 100 A 18
244 1.80 99.6 B 18
245 2.02 100 A 4
246 2.54 99.5 B 4
247 1.11 99.95 A 16
248 3.20 100 B 16
251 0.78 100 A 21
252 1.23 99.95 B 21
253 0.99 100 A 23
254 1.82 99.2 B 23
137 0.98 100 A 12
138 1.33 99.81 B 12
147 1.99 100 B 5
146 1.46 99.18 A 5
154 1.15 100 A 3
155 1.35 99.14 B 3
193 1.16 100 A 4
162 1.44 99.04 B 4
158 1.16 100 A 24
159 1.66 100 B 24
166 1.07 100 A 9
167 1.31 99.03 B 9
173 1.22 100 A 20
174 1.81 100 B 20
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Isomer
Co. Rt SFCMS
UV% Area elution
No. (min) Method
order
168 2.15 99.17 B 10
169 1.21 100 A 10
179 0.90 100 A 11
180 1.29 99.58 B 11
184 1.20 100 A 37
288 1.40 98.50 B 37
185 1.34 100 A 25
187 1.58 100 B 25
295 1.04 100 A 5
299 1.29 98.11 B 5
296 1.40 100 A 9
297 1.88 100 B 9
290 1.42 100 A 37
289 1.57 100 B 37
291 3.50 99.92 A 38
292 3.79 100 B 38
293 1.24 100 A 39
294 1.58 100 B 39
304a 1.40 100 A 40
304b 1.92 99.02 B 40
* Isomer elution order Co. No. 235 vs Co. No. 236; Co. No. 237 vs Co. No. 238.
SFC-MS was also measured for compounds 188, 189, 190 and 191 under the same
SFCMS conditions. The results are shown in Table 2c. (Isomer elution order 'A'
before
13', 13' before 'C', 'C' before 13')
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Co. Rt IsomerSFCMS
UV% Area elution
No. (min) Method
order
188 2.78 100 B 8
189 3.02 96.66 C 8
190 2.40 100 A 8
191 3.21 99.15 D 8
OPTICAL ROTATION (OR)
Optical Rotation is measured with a polarimeter 341 Perkin Elmer. The
polarized light
is passed through a sample with a path length of 1 decimeter and a sample
concentration
of 0.2 to 0.4 gram per 100 milliliters. 2 to 4 mg of the product in vial are
weight, then
dissolved with 1 to 1.2 ml of spectroscopy solvent (DMF for example). The cell
is filled
with the solution and put into the polarimeter at a temperature of 20 C. The
OR is read
with 0.004 of precision.
Calculation of the concentration: weight in gram x 100/ volume in ml
Specific rotation (OR): [a] d20
: (read rotation x 100) / (1.000 dm x concentration).
d is sodium D line (589 nanometer).
Table 3. OR data: wavelength: 589 nm (specicied if different); solvent: DMF
(specicied
if different); temperature: 20 C; 'cone' means concentration (g/100 mL); 'OR'
means
optical rotation.
Co. Wavelength Solvent
OR ( ) Conc.
No. (nm)
20a -31.43 0.28 589 DMF
20b +26.25 0.32 589 DMF
21a -38.48 0.33 589 DMF
21b +42.02 0.326 589 DMF
112 +23.11 0.251 589 DMF
86 -29.55 0.264 589 DMF
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Co. Wavelength Solvent
OR ( ) Conc.
No. (nm)
87 -26.22 0.267 589 DMF
108 -29.61 0.206 589 DMF
109 +25.37 0.272 589 DMF
198 -28.25 0.308 589 DMF
199 +20.91 0.263 589 DMF
196 +8.92 0.269 589 DMF
200 -11.54 0.26 589 DMF
201 +7.81 0.269 589 DMF
202 -5.38 0.26 589 DMF
203 -6.22 0.225 365 DMF
211 +5.65 0.248 589 DMF
213 -9.89 0.263 589 DMF
216 -19.57 0.281 589 DMF
220 -22.63 0.243 589 DMF
221 +11.58 0.259 589 DMF
235 -12.02 0.258 589 DMF
236 +8.58 0.233 589 DMF
237 +31.58 0.228 365 DMF
238 -32.09 0.215 365 DMF
243 -30.12 0.332 365 DMF
244 +20.94 0.277 365 DMF
245 -17.12 0.292 589 DMF
247 -17.25 0.255 365 DMF
248 +5.86 0.239 365 DMF
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Co. Wavelength Solvent
OR ( ) Conc.
No. (nm)
251 -20.6 0.267 589 DMF
252 +12 0.275 589 DMF
253 -25.4 0.252 589 DMF
254 +17.92 0.318 589 DMF
273 +10.38 0.26 589 DMF
274 -5.41 0.222 589 DMF
137 +29.32 0.249 589 DMF
138 -24.9 0.261 589 DMF
147 +8.06 0.273 589 DMF
146 -12.65 0.245 589 DMF
154 -8.59 0.256 589 DMF
155 +6.64 0.241 589 DMF
150 +14.1 0.234 365 DMF
193 -8.62 0.232 365 DMF
162 -5.45 0.275 589 DMF
158 +16.54 0.26 365 DMF
159 -22.31 0.251 365 DMF
166 -12.6 0.254 589 DMF
167 +7.97 0.276 589 DMF
173 +17.92 0.279 365 DMF
174 -21.99 0.282 365 DMF
168 -4.59 0.283 365 DMF
179 +20.19 0.208 365 DMF
180 -93.02 0.215 365 DMF
184 +17.31 0.219 589 DMF
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Co. Wavelength Solvent
OR ( ) Conc.
No. (nm)
288 -26.63 0.338 589 Me0H
185 +32.45 0.256 589 DMF
187 -32.45 0.228 589 DMF
289 +36.73 0.245 589 DMF
290 -41.87 0.246 589 DMF
291 -37.05 0.278 589 DMF
292 +32.39 0.247 589 DMF
293 +29.15 0.295 589 DMF
294 -30.83 0.253 589 DMF
295 +27.38 0.263 589 DMF
296 +50.44 0.113 589 DMF
297 -32.45 0.265 589 DMF
299 -33.79 0.29 589 DMF
304a +8.42 0.285 589 DMF
304b -6.49 0.385 589 DMF
NMR
NMR experiments were carried out using a Bruker Avance 500 spectrometer
equipped
with a Bruker 5mm BBFO probe head with z gradients and operating at 500 MHz
for the
proton and 125 MHz for carbon, or using a Bruker Avance DRX 400 spectrometer
using
internal deuterium lock and equipped with reverse double-resonance (1H, 13C,
SEI) probe
head with z gradients and operating at 400 MHz for the proton and 100MHz for
carbon.
Chemical shifts (6) are reported in parts per million (ppm). J values are
expressed in Hz.
Alternatively, some NMR experiments were carried out using a Bruker Avance III
400
.. spectrometer at ambient temperature (298.6 K), using internal deuterium
lock and
equipped with 5 mm PABBO BB- probe head with z gradients and operating at 400
MHz for the proton and 100MHz for carbon. Chemical shifts (6) are reported in
parts
per million (ppm). J values are expressed in Hz.
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Compound 13:
1H NMR (400MHz, CHLOROFORM-d) 6 ppm 8.42 (s, 1H), 7.38 (s, 1H), 3.95 (br s,
2H), 3.83 (br t, J=6.6 Hz, 2H), 3.63 (q, J=10.1 Hz, 2H), 3.27 (br s, 4H), 2.32
(d, J=6.8
Hz, 2H), 2.28 - 2.18 (m, 2H), 1.69 - 1.57 (m, 1H), 0.90 (d, J=6.6 Hz, 6H)
Compound 20b:
1H NMR (500 MHz, DMSO-d6) 6 ppm 11.64 (br s, 1H) 8.49 (s, 1H) 7.77 (s, 1H)
7.67
(br d, J=3.8 Hz, 2H) 7.35 - 7.51 (m, 3H) 4.32 - 4.48 (m, 2H) 4.03 - 4.18 (m,
2H) 3.91
(br s, 4H) 3.17 - 3.61 (m, 4H) 2.00 - 2.32 (m, 4H)
Compound 82:
1H NMR (500 MHz, DMSO-d6) 6 ppm 8.35 (s, 1H) 7.39 (s, 1H) 4.28 - 4.77 (m, 6H)
4.01 - 4.19 (m, 4H) 3.62 (br t, J=7.9 Hz, 1H) 2.88 (dd, J=9.9, 6.8 Hz, 1H)
2.45 (br dd,
J=9.9, 7.7 Hz, 1H) 2.00 - 2.21 (m, 2H) 1.17 - 1.28 (m, 1H) 0.95 (d, J=6.3 Hz,
3H)
Compound 84:
1H NMR (500 MHz, DMSO-d6) 6 ppm 8.33 (s, 1H) 7.72 (s, 1H) 7.22 (dd, J=8.4, 5.8
Hz, 2H) 7.09 (t, J=9.0 Hz, 2H) 4.08 (q, J=11.0 Hz, 2H) 3.66 - 3.96 (m, 4H)
3.17 (s, 4H)
2.65 - 2.73 (m, 1H) 2.38 - 2.45 (m, 1H) 2.27 (dd, J=12.9, 8.5 Hz, 1H) 2.14 (br
s, 2H)
0.72 (d, J=6.3 Hz, 3H)
Compound 193:
1H NMR (400 MHz, DMSO-d6) 6 ppm 8.32 (s, 1H) 7.70 (s, 1H) 4.07 (q, J=11.1 Hz,
2H) 3.65 -3.91 (m, 6H) 3.07 -3.30 (m, 6H) 2.14 (br s, 2H) 1.85 (br s, 1H) 1.70
- 1.82
(m, 1H) 1.63 (br s, 1H) 1.28 - 1.52 (m, 4H) 0.88 (dd, J=9.6, 7.1 Hz, 6H)
Compound 196:
1H NMR (500 MHz, DMSO-d6) 6 ppm 8.32 (s, 1H) 7.70 (s, 1H) 4.07 (q, J=10.9 Hz,
2H) 3.52 - 3.97 (m, 4H) 3.34 - 3.43 (m, 2H) 3.02 - 3.22 (m, 4H) 2.72 (dd,
J=8.5, 6.3
Hz, 1H) 2.58 (dd, J=8.8, 6.6 Hz, 1H) 2.51 - 2.53 (m, 1H) 2.28 - 2.35 (m, 1H)
2.19 (dd,
J=9.5, 2.2 Hz, 1H) 2.12 (s, 4H) 1.58 (td, J=7.1, 2.2 Hz, 1H) 0.79 (dd, J=14.8,
6.9 Hz,
6H)
Compound 273:
1H NMR (500 MHz, DMSO-d6) 6 ppm 8.32 (s, 1H) 7.67 (s, 1H) 4.03 (q, J=11.0 Hz,
2H) 3.87 (s, 2H) 3.79 (t, J=6.9 Hz, 2H) 3.61 (br t, J=7.6 Hz, 1H) 3.46 (br t,
J=7.9 Hz,
1H) 3.12 - 3.34 (m, 7H) 2.66 - 2.77 (m, 1H) 2.28 (dd, J=9.1, 2.2 Hz, 1H) 2.15
(t, J=6.9
Hz, 2H) 1.62 (td, J=6.9, 2.2 Hz, 1H) 0.82 (dd, J=14.2, 6.9 Hz, 6H)
Compound 274:
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1H NMR (500 MHz, DMSO-d6) 6 ppm 8.31 (s, 1H) 7.69 (s, 1H) 7.45 (s, 1H) 7.20
(s,
1H) 4.07 (q, J=10.9 Hz, 2H) 3.58 - 3.93 (m, 7H) 2.97 - 3.08 (m, 4H) 2.52 -
2.55 (m,
1H) 2.12 (br s, 2H) 1.7 (td, J=6.6, 4.4 Hz, 1H) 0.70 (dd, J=12.6, 6.6 Hz, 6H)
Compound 320:
1HNMR (400MHz , CD30D) 6 8.37 (s,1H), 7.63 (s,1H),7.28 (t, J= 8.8 Hz, 1H),
6.87-
6.86 (m, 3H), 4.30-4.18 (m, 5H), 4.08-3.93 (m, 3H), 3.91-3.88 (m, 2H), 3.88-
3.85 (s,
3H), 3.72-3.70 (m, 1H), 3.06 (dd, J= 5.2Hz, J= 13.6 Hz, 1H), 2.64 (dd, J= 9.2
Hz, J
= 13.6 Hz, 1H), 2.47-2.385 (m, 2H), 1.19-1.18 (d, J = 6.8 Hz 3H)
Compound 321:
1H NMR (400 MHz, DMSO) 6 8.38 (d, J= 5.2 Hz 1H), 7.65 (s, 1H), 7.21 (d, J= 8.4
Hz, 2H), 6.92 (d, J= 8.4 Hz, 2H), 3.75-4.33 (m, 11H), 3.74 (s, 3H), 3.61 (s,
1H), 2.93
(dd, J= 3.6 Hz, 12.8 Hz, 1H), 2.27-2.36 (m, 2H,), 1.03 (d, J=5.6 Hz, 3H).
Compound 322:
1H NMR (400 MHz, CD30D) 6 8.40 (s, 1H), 7.66 (s, 1H), 7.38-7.34 (m, 2H), 7.22-
1.13
(m, 2H), 4.37-4.26 (m, 5H), 4.13 (s, 1H), 4.01 (s, 2H), 3.91 (q, J=10.4Hz,
2H), 3.74-
3.69 (m, 1H), 2.98 (dd, J= 3.6 Hz, 13.2 Hz 1H), 2.68 (dd, J= 9.6 Hz, 13.2 Hz
1H),
2.47 (s, 2H), 1.20 (d, J= 6.4 Hz 3H).
Compound 323:
1FINMR (400 MHz, CD30D) 6 8.38 (s, 1H), 7.64 (s, 1H), 7.10 (d, J= 8.4 Hz, 2H),
6.78(d, J=8.4 Hz, 2H), 3.86-7.27 (m, 10H), 3.60-6.62 (m, 1H), 2.96 (dd, J=4.8
Hz,14.0Hz, 1H), 2.60 (dd, J = 4.8 Hz, 14.0Hz 1H), 2.37-2.46 (m, 2H), 1.18 (d,
J=
6.8Hz, 3H).
Compound 324:
1H NMR (400 MHz, CD30D) 6 8.36 (s,1H),7.62 (s,1H),7.19-7.15 (m, 4H), 4.28-4.16
(m, 5H), 4.14-3.98 (m, 4H), 3.93-3.87 (m, 2H), 3.66-3.64 (m, 1H), 3.04 (dd, J=
4.8
Hz, J= 13.6 Hz, 1H), 2.63 (dd, J= 9.2 Hz, J= 13.6 Hz 1H), 2.46-2.32 (m, 5H),
1.18
(d, J= 6.4Hz, 3H).
PHARMACOLOGICAL PART
1) Menin/MLL fluorescence polarization assay
To a non-surface binding, black 384-well microtiter plate was added 50 nL 160X
test
compound in DMSO and 4 iut 2X menin in assay buffer (40 mM Tris=FIC1, pH 7.5,
50
mM NaCl, 1 mM DTT and 0.001% Tween 20). After incubation of test compound and
menin for 10 min at ambient temperature, 4 iut 2X FITC-MBM1 peptide (FITC-13-
alanine-SARWRFPARPGT-NH2) in assay buffer was added, the microtiter plate
centrifuged at 1000 rpm for 1 min and the assay mixtures incubated for 15 min
at
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ambient temperature. The relative amount of menin=FITC-MBM1 complex present in
an assay mixture is determined by measuring the fluorescence polarization (FP)
of the
FITC label with a BMG Pherastar plate reader (ex. 485 nm/em. 520 nm) at
ambient
temperature. The final concentrations of reagents in the binding assay are 100
nM
.. menin, 5 nM FITC-MBM1 peptide and 0.625% DMSO in assay buffer. Dose-
response
titrations of test compounds are conducted using an 11 point, three-fold
serial dilution
scheme, starting at 31 M.
Compound potencies were determined by first calculating % inhibition at each
compound concentration according to equation 1:
% inhibition = ((HC - LC) - (FPcompound _ LC)) /(HC - LC)) *100 (Eqn 1)
Where LC and HC are the FP values of the assay in the presence or absence of a
saturating concentration of a compound that competes with FITC-MBM1 for
binding to
menin, and FP'mP und is the measured FP value in the presence of the test
compound.
HC and LC FP values represent an average of at least 16 replicates per plate.
For each
.. test compound, % inhibition values were plotted vs. the logarithm of the
test compound
concentration, and the /C50 value derived from fitting these data to equation
2:
% inhibition = Bottom + (Top-Bottom)/(1+10^((log/C50-log[cmpd])*h)) (Eqn 2)
Where Bottom and Top are the lower and upper asymptotes of the dose-response
curve,
respectively, /C50 is the concentration of compound that yields 50% inhibition
of signal
and h is the Hill coefficient.
2) Proliferation assay
The anti-proliferative effect of menin/MLL protein/protein interaction
inhibitor test
compounds was assessed in human leukemia cell lines. The cell lines MV-4-11
and
MOLM14 harbor MLL translocations and express the MLL fusion proteins MLL-AF4
and MLL-AF9, respectively, as well as the wildtype protein from the second
allele.
Therefore, the MLL rearranged cell lines MV-4-11 and MOLM14 exhibit stem cell-
like
HOXAIMEIS1 gene expression signatures. K562 and KG1 were used as a control
cell
lines containing two MLL wildtype alleles in order to exclude compounds that
display
general cytotoxic effects.
MV-4-11 and MOLM14 were cultured in RPMI-1640 (Sigma Aldrich) supplemented
with 10% fetal bovine serum (HyClone), 2 mM L-glutamine (Sigma Aldrich) and
50 g/m1 gentamycin (Gibco). K562 were propagated in RPMI-1640 (Sigma Aldrich)
supplemented with 20% fetal bovine serum (HyClone), 2 mM L-glutamine (Sigma
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Aldrich) and 50 g/m1 gentamycin (Gibco). KG1 were cultured in Iscove's MDM
(Gibco) supplemented with 20% fetal bovine serum (HyClone), 2 mM L-glutamine
(Sigma Aldrich) and 50 g/m1 gentamycin (Gibco). Cells were kept at 0.3 ¨ 2.5
million
cells per ml during culturing and passage numbers did not exceed 25.
.. In order to assess the anti-proliferative effects, 1,500 MV-4-11, 300
MOLM14, 750
K562 or 1,300 KG1 cells were seeded in 200 1 media per well in 96-well round
bottom, ultra-low attachment plates (Costar, catalogue number 7007). Cell
seeding
numbers were chosen based on growth curves to ensure linear growth throughout
the
experiment. Test compounds were added at different concentrations and the DMSO
content was normalized to 0.3%. Cells were incubated for 8d at 37 C and 5%
CO2.
Spheroid like growth was monitored in real-time by live-cell imaging
(IncuCyteZOOM, Essenbio, 4x objective) acquiring one image every four hours
for 8d.
Confluence (%) as a measure of spheroid size was determined using an
integrated
analysis tool.
In order to determine the cumulative effect of the test compounds over time,
the area
under the curve (AUC) in a plot of confluence against time was calculated.
Confluence
at the beginning of the experiment (t=0) was used as baseline for the AUC
calculation.
Absolute IC50 values were calculated according to the following procedure:
%Control = (AUC sample/AUC control)*100
AUC control = mean AUC of control values (cells without compound/DMSO as
vehicle control)
A non-linear curve fit was applied using the least squares (ordinary) fit
method to the
plot of % control versus compound concentration. Based on this, the absolute
ICso
value (half maximal inhibitory concentration of the test compound causing an
anti-
.. proliferative effect of 50% relative to the vehicle control) was
calculated.
3) Menin/MLL homogenous time-resolved fluorescence (HTRF) assay
To an untreated, white 384-well microtiter plate was added 40 nL 200X test
compound
in DMSO and 4 iut 2X terbium chelate-labeled menin (vide infra for
preparation) in
assay buffer (40 mM Tris=HC1, pH 7.5, 50 mM NaCl, 1 mM DTT and 0.05% Pluronic
F-127). After incubation of test compound and terbium chelate-labeled menin
for 5
min at ambient temperature, 4 iut 2X FITC-MBM1 peptide (FITC-13-alanine-
SARWRFPARPGT-NH2) in assay buffer was added, the microtiter plate centrifuged
at
1000 rpm for 1 min and the assay mixtures incubated for 15 min at ambient
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temperature. The relative amount of menin=FITC-MBM1 complex present in an
assay
mixture is determined by measuring the homogenous time-resolved fluorescence
(HTRF) of the terbium/FITC donor /acceptor fluorphore pair using a BMG
Pherastar
plate reader (ex. 337 nm/terbium em. 490 nm/FITC em. 520 nm) at ambient
temperature. The degree of fluorescence resonance energy transfer (the HTRF
value) is
expressed as the ratio of the fluorescence emission intensities of the FITC
and terbium
fluorophores (Pm 520 nm/Fm 490 nm). The final concentrations of reagents in
the
binding assay are 100 pM terbium chelate-labeled menin, 75 nM FITC-MBM1
peptide
and 0.5% DMSO in assay buffer. Dose-response titrations of test compounds are
conducted using an 11 point, three-fold serial dilution scheme, starting at 31
M.
Compound potencies were determined by first calculating % inhibition at each
compound concentration according to equation 1:
% inhibition = ((HC - LC) _ (HTRpompound _ LC)) / (HC - LC)) *100 (Eqn 1)
Where LC and HC are the HTRF values of the assay in the presence or absence of
a
saturating concentration of a compound that competes with FITC-MBM1 for
binding to
menin, and HTRFc 111P 1111d is the measured HTRF value in the presence of the
test
compound. HC and LC HTRF values represent an average of at least 16 replicates
per
plate. For each test compound, % inhibition values were plotted vs. the
logarithm of
the test compound concentration, and the /C50 value derived from fitting these
data to
equation 2:
% inhibition = Bottom + (Top-Bottom)/(1+10^((log/C50-log[cmpd])*h)) (Eqn 2)
Where Bottom and Top are the lower and upper asymptotes of the dose-response
curve,
respectively, /C50 is the concentration of compound that yields 50% inhibition
of signal
and h is the Hill coefficient.
Preparation of Terbium cryptate labeling of Menin: Menin (a.a. 1-610-6xhis
tag) was
labeled with terbium cryptate as follows. 2mg of Menin was buffer exchanged
into lx
phosphate buffered saline. 16uM Menin was incubated with 4-fold molar excess
NHS-
terbium cryptate (Cisbio Bioassays, Bedford, MA) for 2 hours at room
temperature.
The labeled protein was purified away from free label by running the reaction
over a
Superdex 200 Increase 10/300 GL column at 0.75m1/min. Peak fractions were
collected, aliquoted and frozen at -80 C.
MENIN Protein Sequence (SEQ ID NO: 1):
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MGLKAAQKTLFPLRSIDDVVRLFAAELGREEPDLVLLSLVLGFVEHFLAVNRVI
PTNVPELTFQPSPAPDPPGGLTYFPVADL SIIAALYARFTAQIRGAVDLSLYPRE
GGVS SRELVKKVSDVIWNSLSRSYFKDRAHIQSLFSFITGTKLDS SGVAFAVVG
AC QALGLRDVHLAL SEDHAWVVFGPNGEQTAEVTWHGKGNEDRRGQTVNA
GVAERSWLYLKGSYMRCDRKMEVAFMVCAINPSIDLHTDSLELLQLQQKLLW
LLYDLGHLERYPMALGNLADLEELEPTPGRPDPLTLYHKGIASAKTYYRDEHI
YPYMYLAGYHCRNRNVREALQAWADTATVIQDYNYCREDEEIYKEFFEVAN
DVIPNLLKEAASLLEAGEERPGEQSQGTQSQGSALQDPECFAHLLRFYDGICKW
EEG S PTPVLHVGWATFLVQ S LGRFE GQVRQKVRIV S REAEAAEAEEPWGEEAR
EGRRRGPRRESKPEEPPPPKKPALDKGLGTGQGAVSGPPRKPPGTVAGTARGPE
GGSTAQVPAPAASPPPEGPVLTFQSEKMKGMKELLVATKINS SAIKLQLTAQSQ
VQMKKQKVSTPSDYTLSFLKRQRKGLHHHHHH
Table 4. Biological data in the Menin fluorescence polarization (FP) assay
(1),
Menin/MLL homogenous time-resolved fluorescence (HTRF) assay (3) and
proliferation
assay (2). Co. No. means compound number. The values in table 4 are averaged
values
over all measurements.
(2) (2) Spheroid (2) (2)
(1) (3) Spheroid assay Spheroid Spheroid
Menin Menin
assay MOLM14 assay K562 assay KG!
Co. FP HTRF
MV-4-11 (IC50 ( M)) (IC50( M)) (IC5o( M))
No. assay assay
(IC50
(IC50 (IC50 (104))
(104)) (nM))
18 0.14 227 3.9 7.7 >15
1 0.056 144 3.7 4.6 >15
3 0.10 216 3.8 5.6 ¨15
14 0.11 178 2.2 4.9
20a 0.076 13 0.96 5 >15
21a 0.49 745 6.9 12.1 >15
21b 0.13 389 4.2 4.6 >15
19 0.81 1960
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(2) (2) Spheroid (2) (2)
(1) (3) Spheroid assay Spheroid Spheroid
Menin Menin
assay MOLM14 assay K562 assay KG!
Co. FP HTRF
MV-4-11 (IC50 ( M)) (IC50( M)) (IC5o( M))
No. assay assay (IC50
(IC50 (IC50 (I1M))
(I1M)) (104))
0.67 600 6.4 30.6 >15
12 0.63 2247 >15 >15 >15
13 0.042 39 0.73 4.2 >15
11 0.76 2374
2.31
9 0.84 1579
8 0.79 2525
6 0.24 339 2.4 10.1 >15
7 0.73 1216
4 0.73 3164
2 0.39 706 4.9 14.6 >15
0.57 471 14.0 >15 >15
22 0.069 12 0.55 2.0 >15
23 0.95 1208
24 0.49 1486 9.1 >15
0.31 1465 3.8 7.2
26 0.16 143 10.4 >15
197 0.30 541 7.3 >15
86 0.092 102 2.6 11.3 >15
133 0.12 385 2.1 >15
0.53 839 9.2 >15
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(2) (2) Spheroid (2) (2)
(1) (3) Spheroid assay Spheroid Spheroid
Menin Menin
assay MOLM14 assay K562 assay KG!
Co. FP HTRF
MV-4-11 (IC50 ( M)) (IC50( M)) (IC5o( M))
No. assay assay (IC50
(IC50 (IC50 (I1M))
(I1M)) (nM))
88 0.32 486 >15 >15
89 0.045 31 1.3 10.5
90 0.11 288 11.1 >15
74 0.12 211 2.0 6.1
75 2.36
34 0.72 902
198 0.14 178 5.1 12.4
199 0.41 1196 12.9 >15
35 0.99 2375
36 0.4 777
37 0.17 177 1.5 3.1
76 0.68 901
40 0.84 1465
195 1.07 1536
41 0.63 950
273 0.021 8 0.08 0.86 10.3 12
200 0.11 190 3.4 10.3
201 1075 12.7 >15
202 490 >15
137 1097 10.1 >15
44 344 4.1 >15
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(2) (2) Spheroid (2) (2)
(1) (3) Spheroid assay Spheroid Spheroid
Menin Menin
assay MOLM14 assay K562 assay KG!
Co. FP HTRF
MV-4-11 (IC50 ( M)) (IC50( M)) (IC5o( M))
No. assay assay (IC50
(IC50 (IC50 (I1M))
(I1M)) (nM))
82 9 0.38 1.8 >15 >15
105 15 0.67 3.4 >15
108 753 >15
109 571 8.1
45 330 3.4
46 448 4.1
47 669
48 483 2.0
49 310 2.1
50 665
110 236 4.0
111 394 9.3
77 191 12.1
54 342 3.7
56 304 4.9
112 1540
274 7 0.36 2.3 >15 >15
203 422 7.2
145 37 1.7 7.3
146 46 2.4
147 341 8.2
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(2) (2) Spheroid (2) (2)
(1) (3) Spheroid assay Spheroid Spheroid
Menin Menin
assay MOLM14 assay K562 assay KG!
Co. FP HTRF
MV-4-11 (IC50 ( M)) (IC50( M)) (IC5o( M))
No. assay assay (IC50
(IC50 (IC50 (I1M))
(I1M)) (nM))
57 179 4.2 8.6
148 5 0.33
211 13 0.68 3.5 12.5
213 149 1.9
214 17 0.76 1.4 >15
150 60 2.0
220 75 0.65 4.7 >15 >15
221 1157
222 4 0.2
223 496 6.1
228 238
58 185 2.7
59 1245
230 44
231 107
232 914
233 782
234 1045
235 23 0.71
236 1041
237 52 1.2
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(2) (2) Spheroid (2) (2)
(1) (3) Spheroid assay Spheroid Spheroid
Menin Menin
assay MOLM14 assay K562 assay KG!
Co. FP HTRF
MV-4-11 (IC50 ( M)) (IC50( M)) (IC5o( M))
No. assay assay (IC50
(IC50 (IC50 (I1M))
(I1M)) (nM))
238 659
154 328
155 69 0.7 1.2 >15
243 249
244 848
245 10 0.32 2 >15
246 284
247 19 0.45 2.1 >15 >15
248 549
61 513
249 912
250 190
251 1660
252 1007
253 323
254 861
156 21 0.79
157 101
158 281
159 1174
62 28 0.64 4 >15
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(2) (2) Spheroid (2) (2)
(1) (3) Spheroid assay Spheroid Spheroid
Menin Menin
assay MOLM14 assay K562 assay KG!
Co. FP HTRF
MV-4-11 (IC50 ( M)) (IC50( M)) (IC5o( M))
No. assay assay (IC50
(IC50 (IC50 (I1M))
(I1M)) (nM))
161 187 4.2
258 3083
193 25 0.65 1.7 >15 >15
162 77 3.1
63 201
196 4 0.3 1.1 >15 >15
163 277
164 1125
259 20 1.3
64 1139
65 65 3.2
166 79 2.4
167 1126
168 3 0.19 0.86 11.9 15
169 176 3.5
170 138 4.3
84 20 0.58 1.5 >15
171 159 3.1
66 26 0.95 0.69 >15
172 124 2.9
173 96 3.2
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(2) (2) Spheroid (2) (2)
(1) (3) Spheroid assay Spheroid Spheroid
Menin Menin
assay MOLM14 assay K562 assay KG!
Co. FP HTRF
MV-4-11 (IC50 ( M)) (IC50( M)) (IC5o( M))
No. assay assay (IC50
(IC50 (IC50 (I1M))
(I1M)) (nM))
174 1386
67 108 4.9
179 95 1.3
180 130 1.9
69 186 1.2
182 61 0.78 1.9 >15
183 103 1.3
71 48 1.1
184 73 0.97 >15
186 91 1.3
187 72 1.4
269 8 0.67 4.6 >15
188 115 1.5
189 108 0.92 >15
270 499
271 623
190 524
191 1263
192 619
312 339
313 159 3.6
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(2) (2) Spheroid (2) (2)
(1) (3) Spheroid assay Spheroid Spheroid
Menin Menin
assay MOLM14 assay K562 assay KG!
Co. FP HTRF
MV-4-11 (IC50 ( M)) (IC50( M)) (IC5o( M))
No. assay assay (IC50
(IC50 (IC50 (I1M))
(I1M)) (nM))
315 21 0.32 1.5 >15
316 20 0.4 0.96 >3.7
288 68 1.4
289 6 0.21 1.3
290 25 0.47 1.2
291 13 0.66 1.2 >15
292 12 0.29 1.5 >15
294 82 1.83
293 81 1.67
307 11 0.28 1.5 10.7
299 57 3.18
295 34 1.16 6.7
303 6 0.32 1.02
296 200 4.89
309 16 0.74 1.6
297 242
304b 440
304a 15 0.44 1.3 >15
317 248 0.77 >15
318 72 1.09
298 122 4.23
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(2) (2) Spheroid (2) (2)
(1) (3) Spheroid assay Spheroid Spheroid
Menin Menin
assay MOLM14 assay K562 assay KG!
Co. FP HTRF
MV-4-11 (IC50 ( M)) (IC50( M)) (IC5o( M))
No. assay assay
(ICso
(IC50 (IC50 (104))
(I1M)) (04))
314 25 0.73
320 259 1.37
321 210 1.1
322 200 1.7
323 47 0.75
324 143 1.9
31 0.087 116 1.3 9.8
224 2365
185 161 1.3
311 85 2.4
Table 5. Biological data in the Menin fluorescence polarization (FP) assay
(1),
Menin/MLL homogenous time-resolved fluorescence (HTRF) assay (3) and
proliferation
assay (2). Co. No. means compound number. The values in table 5 are values for
individual measurements (not averaged): in case a value was determined more
than 1
time, each value is reported individually in Table 5.
(2) (2) Spheroid (2) (2)
(1) (3) Spheroid assay Spheroid Spheroid
Menin Menin
assay MOLM14 assay K562 assay KG!
Co. FP HTRF
MV-4-11 (IC50 ( M)) (IC50( M)) (IC5o( M))
No. assay assay
(ICso
(IC50 (IC50 (104))
(I1M)) (04))
20b 0.038 7 0.51 >15 >15 >15
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(2) (2) Spheroid (2) (2)
(1) (3) Spheroid assay Spheroid Spheroid
Menin Menin
assay MOLM14 assay K562 assay KG!
Co. FP HTRF
MV-4-11 (IC50 ( M)) (IC50( M)) (IC5o( M))
No. assay assay (IC50
(IC50 (IC50 (I1M))
(I1M)) (nM))
2.5
4.7 9.3
16
0.134 63 >15 8.7 >15
4.9
17 0.76 3.4
0.054 14 0.84 2.3 11
0.5
28 10.8 3.2
0.517 1262 10.9 >15
1.8
29 >15 10.8
0.346 1438 9.9 >15
4.1
0.63
0.58
87
0.63 8.0
0.68 2.8
0.048 28 0.52 4.6 >15
¨15.4
138 1.0
62 1.4 7
