Note: Descriptions are shown in the official language in which they were submitted.
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Substituted oxopyridine derivatives
The invention relates to substituted oxopyridine derivatives and to processes
for their preparation,
and also to their use for preparing medicaments for the treatment and/or
prophylaxis of diseases, in
particular vascular disorders, preferably thrombotic or thromboembolic
disorders and/or thrombotic
or thromboembolic complications.
Haemostasis is a protective mechanism of the organism, which helps to "seal"
leaking damages in
the blood vessel wall quickly and reliably. Thus, excessive loss of blood can
often be avoided or kept
to a minimum. After injury of a blood vessel, hemostasis is conducted mainly
by activation and
aggregation of platelets and activation the coagulation system, which consists
of an enzymatic
"waterfall" cascade leading one after another to the activation of the next
coagulation factor until
thrombin is formed, which leads to the generation of insoluble fibrin, which
is an important part of
the clot.
In the more recent past, the traditional theory of two separate starting
points of the coagulation
cascade (extrinsic and intrinsic path) has been modified owing to new
findings: In these models,
coagulation is initiated by binding of activated factor VIIa to tissue factor
(TF). The resulting
complex activates factor X, which in turn leads to generation of thrombin with
subsequent production
of fibrin and platelet activation (via PAR-1) as injury-sealing end products
of haemostasis. Compared
to the subsequent amplification/propagation phase, the thrombin production
rate in this first phase is
low and as a result of the occurrence of TFPI as inhibitor of the TF-FVIIa-FX
complex is limited in
time. A central component of the transition from initiation to amplification
of coagulation and
thereby thrombus propagation is factor XIa: in positive feedback loops,
thrombin activates not only
factor V and factor VIII, but also factor XI to factor XIa, which in turn
converts factor IX into factor
IXa, which in turn in a factor IXa/factor Villa complex generates factor Xa
and finally to large
amounts of thrombin, resulting in strong thrombus growth and stabilization of
the thrombus. This is
supported by TAFIa and FXIIIa, which are activated by thrombin as well and
lead to inhibition of
clot lysis and further clot stabilisation.
In addition to the stimulation via tissue factor, the coagulation system can
be activated particularly
on negatively charged surfaces, which include not only surface structures of
foreign cells (e.g.
bacteria) but also artificial surfaces such as vascular prostheses, stents and
extracoporeal circulation.
On these surfaces, factor XII (FXII) is activated to factor XIIa, which
subsequently activates factor
XI to factor XIa. This leads to further activation of the coagulation cascade
as described above. In
addition, factor XIIa also activates bound plasma prokallikrein to plasma
kallikrein (PK) which, in a
potentiation loop, firstly leads to further factor XII activation, overall
resulting in amplification of
the initiation of this intrinsic part of the coagulation cascade.
Uncontrolled activation of the coagulation system or defective inhibition of
the activation processes
may lead to the formation of local thrombi or emboli in vessels (e.g.
arteries, veins, lymph vessels)
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or in organ cavities (e.g. cardiac atrium). In addition, systemic
hypercoagulability may lead to
system-wide formation of microthrombi and finally to a consumption
coagulopathy in the context of
a disseminated intravasal coagulation. Thromboembolic complications may also
occur in
extracorporeal circulatory systems, such as haemodialysis, and also in
vascular prostheses or
prosthetic heart valves and stents.
In the course of many cardiovascular and metabolic disorders, increased
tendency for coagulation and
platelet activation occur owing to either systemic factors such as
hyperlipidaemia, diabetes,
inflammation, infection or smoking, or to changes in blood flow with stasis,
for example in in diseased
leg veins or in atrial fibrillation, or owing to pathological changes in
vessel walls, for example endothelial
dysfunctions or atherosclerosis. This unwanted and excessive activation of
coagulation may, by
formation of fibrin- and platelet-rich thrombi, lead to thromboembolic
disorders and thrombotic
complications with often life-threatening events. Inflammation processes may
also be involved by
triggering the coagulation system. On the other hand, thrombin is known to
activate inflammatory
pathways, as well.
Accordingly, thromboembolic disorders are still the most frequent cause of
morbidity and mortality in
most industrialized countries.
The anticoagulants known from the prior art, that is to say substances for
inhibiting or preventing
blood coagulation, have various disadvantages. Accordingly, in practice,
efficient treatment methods
or the prophylaxis of thrombotic/thromboembolic disorders is found to be
difficult and
unsatisfactory.
In the therapy and prophylaxis of thromboembolic disorders, use is made,
firstly, of heparin which
is administered parenterally or subcutaneously. Because of more favourable
pharmacokinetic
properties, preference is these days increasingly given to low-molecular-
weight heparin; however,
the known disadvantages described herein below encountered in heparin therapy
cannot be avoided
either in this manner. Thus, heparin is orally ineffective and has only a
comparatively short half-life.
In addition, there is a high risk of bleeding, there may in particular be
cerebral haemorrhages and
bleeding in the gastrointestinal tract, and there may be thrombopaenia,
alopecia medicomentosa or
osteoporosis. Low-molecular-weight heparins do have a lower probability of
leading to the
development of heparin-induced thrombocytopaenia; however, they can also only
be administered
subcutaneously. This also applies to fondaparinux, a synthetically produced
selective factor Xa
inhibitor having a long half-life.
A second class of anticoagulants are the vitamin K antagonists. These include,
for example, 1,3-
indanediones and in particular compounds such as warfarin, phenprocoumon,
dicumarol and other
coumarin derivatives which non-selectively inhibit the synthesis of various
products of vitamin K-
dependent coagulation factors in the liver. Owing to the mechanism of action,
the onset of action is
only very slow (latency to the onset of action 36 to 48 hours). The compounds
can be administered
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orally; however, owing to the high risk of bleeding and the narrow therapeutic
index complicated
individual adjustment and monitoring of the patient are required. In addition,
other side-effects such
as gastrointestinal problems, hair loss and skin necroses have been described.
Today, approaches for Non-vitamin K dependent oral anticoagulantion (NOACs)
are in clinical use,
and have demonstrated their effectiveness in various studies. However, taking
these medicaments
can also lead to bleeding complications, particularly in predisposed patients.
Thus, for antithrombotic medicaments, the therapeutic window is of central
importance: The interval
between the therapeutically active dose for coagulation inhibition and the
dose where bleeding may
occur should be as large as possible so that maximum therapeutic activity is
achieved at a minimum
risk profile.
In various in vitro and in vivo models with, for example, antibodies as factor
XIa inhibitors, but also
in factor XIa knock-out animal models, the antithrombotic effect with small/no
prolongation of
bleeding time or extension of blood volume was confirmed. In clinical studies,
elevated factor XIa
concentrations were associated with an increased thrombotic event rate. In
contrast, factor XI
deficiency (haemophilia C) did not lead to spontaneous bleeding and was
apparent only in the course
of surgical operations and traumata, but did show protection with respect to
certain thromboembolic
events.
Furthermore, for many disorders the combination of antithrombotic and
antiinflammtory principles
may also be particularly attractive to prevent the mutual enhancement of
coagulation and
inflammation.
It is therefore an object of the present invention to provide novel compounds
for the treatment of
cardiovascular disorders, in particular of thrombotic or thromboembolic
disorders and/or thrombotic
or thromboembolic complications, in humans and animals, which compounds have a
wide
therapeutic window.
WO 2006/030032 describes inter alia substituted pyridinones as allosteric
modulators of the mGluR2
receptor, and WO 2008/079787 describes substituted pyridin-2-ones and their
use as glucokinase
activators. WO 2014/154794, WO 2014/160592, WO 2015/011087, WO 2015/063093, WO
2016/046158, WO 2016/046157, WO 2016/046159, WO 2016/046164, WO 2016/046166,
WO
2016/046156, WO 2017/005725 and WO 2017/037051 describe substituted pyridin-2-
ones and their
use as factor XIa inhibitors.
The invention provides compounds of the formula
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R2
R3
R4
R 0
N R5
0
0
R8
R6
(I),
R7
CI
in which
RI represents methyl, ethyl, difluoromethyl or trifluoromethyl,
R2 represents hydrogen, methyl, difluoromethyl or trifluoromethyl,
or
RI and R2 together with the carbon atoms to which they are attached form a
cyclobutyl ring,
IV represents methyl, ethyl or n-propyl,
where methyl may be substituted with one substituent selected from the group
consisting of
cyclopropyl, cyclobutyl, oxetan-2-yl, oxetan-3-yl, tetrahydrofuran-2-yl,
tetrahydro-2H-
pyran-2-yl, tetrahydro-2H-pyran-4-y1 and 1,4-dioxan-2-yl,
where oxetan-2-yl, tetrahydrofuran-2-yl, tetrahydro-2H-pyran-2-y1 and 1,4-
dioxan-
2-y1 may be substituted by 1 to 2 substituents independently of one another
selected
from the group consisting of fluorine and methyl,
or
where methyl may be substituted with one sub stituent of the group of the
formula
R10
R10 OH
0' 0'
*R9 or 9 or
L'R
where
is the attachment site to the methyl group,
R9 represents methyl, ethyl, iso-propyl, cyclopropyl,
difluoromethyl or
trifluoromethyl,
RI represents methyl or difluoromethyl,
and
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where ethyl may be substituted with one substituent selected from the group
consisting of
methoxy, ethoxy, iso-propoxy, tert-butoxy, difluoromethoxy, trifluoromethoxy,
2,2-
difluoroethoxy, 2,2,2-trifluoroethoxy, cyclopropyloxy and cyclobutyloxy,
where cyclopropyloxy and cyclobutyloxy may be substituted with one substituent
selected from the group consisting of fluorine and methyl,
R4 represents hydrogen,
R5 represents a group of the formula
R11
# 0 #r),,r N
N H 2 or / N H 2
or #
---. /
N R-12
N
0 0
# N # N
or
D,13 ../...."...sy....--- \ N or # \..,.....,N
N
n,15
or rx
0 ..., / N4
N¨rx
\ 14
R
R17
#.7N #............Nµ #N-4
r\V. or )_R16
N
or or N,...\
N /LNI/
0
0
#
N=sj.4
or I NCH 3
N1/
where
10 # is the attachment site to the nitrogen atom,
RH represents hydrogen or fluorine,
R12 represents methyl, difluoromethyl or trifluoromethyl,
RI' represents methyl, difluoromethyl or trifluoromethyl,
R14 represents hydrogen or methyl,
R15 represents hydrogen or methyl,
R16 represents hydrogen or methyl,
R17 represents hydrogen or methyl,
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R6, R7 and R8 represent the following:
R6 represents hydrogen, fluorine or chlorine,
R7 represents hydrogen,
R8 represents hydrogen,
or
R6 represents hydrogen,
R7 represents fluorine or chlorine,
R8 represents hydrogen,
or
R6 represents hydrogen,
R7 represents hydrogen,
R8 represents fluorine,
and the salts thereof, the solvates thereof and the solvates of the salts
thereof
Compounds according to the invention are the compounds of the formula (I) and
the salts, solvates
and solvates of the salts thereof, and also the compounds encompassed by
formula (I) and specified
hereinafter as working example(s), and the salts, solvates and solvates of the
salts thereof, to the
extent that the compounds encompassed by formula (I) and specified hereinafter
are not already salts,
solvates and solvates of the salts.
The inventive compounds may, depending on their structure, exist in different
stereoisomeric forms,
i.e. in the form of configurational isomers or else, if appropriate, of
conformational isomers
(enantiomers and/or diastereomers, including those in the case of rotamers and
atropisomers). The
present invention therefore encompasses the enantiomers and diastereomers, and
the respective
mixtures thereof The stereoisomerically uniform constituents can be isolated
from such mixtures of
enantiomers and/or diastereomers in a known manner; chromatography processes
are preferably used
.. for this, especially HPLC chromatography on an achiral or chiral phase.
If the compounds according to the invention can occur in tautomeric forms, the
present invention
encompasses all the tautomeric forms.
In the context of the present invention, the term "enantiomerically pure" is
understood to mean that
the compound in question with respect to the absolute configuration of the
chiral centre is present in
an enantiomeric excess of more than 95%, preferably more than 97%. The
enantiomeric excess (ee
value) is calculated in this case by evaluation of the corresponding HPLC
chromatogram on a chiral
phase with the aid of the formula below:
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ee = [EA (area%) - EB (area%)] x 100% / [EA (area%) + EB (area%)]
(EA: enantiomer in excess, EB: enantiomer in deficiency)
The present invention also encompasses all suitable isotopic variants of the
compounds according to
the invention. An isotopic variant of an inventive compound is understood here
as meaning a
.. compound in which at least one atom within the inventive compound has been
exchanged for another
atom of the same atomic number, but with a different atomic mass than the
atomic mass which
usually or predominantly occurs in nature. Examples of isotopes which can be
incorporated into a
compound according to the invention are those of hydrogen, carbon, nitrogen,
oxygen, phosphorus,
sulfur, fluorine, chlorine, bromine and iodine, such as 2H (deuterium), 3H
(tritium), 13C, 14C, 15N, 170,
180, 32F, 33F, 33s, 34s, 35s, 36s, 18F, 36C1, 82Br, 1231, 1241, 1291 and 131J
a I. Particular isotopic variants of a
compound according to the invention, especially those in which one or more
radioactive isotopes
have been incorporated, may be beneficial, for example, for the examination of
the mechanism of
action or of the active ingredient distribution in the body; due to
comparatively easy preparability
and detectability, especially compounds labelled with 3H or 14C isotopes are
suitable for this purpose.
.. In addition, the incorporation of isotopes, for example of deuterium, may
lead to particular
therapeutic benefits as a consequence of greater metabolic stability of the
compound, for example an
extension of the half-life in the body or a reduction in the active dose
required; such modifications
of the inventive compounds may therefore in some cases also constitute a
preferred embodiment of
the present invention. Isotopic variants of the compounds according to the
invention can be prepared
by the processes known to those skilled in the art, for example by the methods
described further
below and the procedures described in the working examples, by using
corresponding isotopic
modifications of the respective reagents and/or starting compounds.
Preferred salts in the context of the present invention are physiologically
acceptable salts of the
compounds according to the invention. However, the invention also encompasses
salts which themselves
.. are unsuitable for pharmaceutical applications but which can be used, for
example, for the isolation or
purification of the compounds according to the invention.
Physiologically acceptable salts of the compounds according to the invention
include acid addition salts
of mineral acids, carboxylic acids and sulfonic acids, for example salts of
hydrochloric acid,
hydrobromic acid, sulfuric acid, phosphoric acid, methanesulfonic acid,
ethanesulfonic acid,
.. toluenesulfonic acid, benzenesulfonic acid, naphthalenedisulfonic acid,
acetic acid, trifluoroacetic acid,
propionic acid, lactic acid, tartaric acid, malic acid, citric acid, fumaric
acid, maleic acid and benzoic
acid.
Physiologically acceptable salts of the compounds according to the invention
also include salts of
conventional bases, by way of example and with preference alkali metal salts
(e.g. sodium and potassium
salts), alkaline earth metal salts (e.g. calcium and magnesium salts) and
ammonium salts derived from
ammonia or organic amines having 1 to 16 carbon atoms, by way of example and
with preference
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ethylamine, diethylamine, triethylamine, ethyldiisopropylamine,
monoethanolamine, diethanolamine,
triethanolamine, dicyclohexylamine, dimethylaminoethanol, procaine,
dibenzylamine, N-
methylmorpholine, arginine, lysine, ethylenediamine, N-methylpiperidine and
choline.
Solvates in the context of the invention are described as those forms of the
inventive compounds which
form a complex in the solid or liquid state by coordination with solvent
molecules. Hydrates are a
specific form of the solvates in which the coordination is with water.
The present invention additionally also encompasses prodrugs of the inventive
compounds. The term
"prodrugs" encompasses compounds which for their part may be biologically
active or inactive but are
converted during their residence time in the body into compounds according to
the invention (for
example by metabolism or hydrolysis).
In the context of the present invention, the term "treatment" or "treating"
includes inhibition,
retardation, checking, alleviating, attenuating, restricting, reducing,
suppressing, repelling or healing
of a disease, a condition, a disorder, an injury or a health problem, or the
development, the course or
the progression of such states and/or the symptoms of such states. The term
"therapy" is understood
here to be synonymous with the term "treatment".
The terms "prevention", "prophylaxis" and "preclusion" are used synonymously
in the context of the
present invention and refer to the avoidance or reduction of the risk of
contracting, experiencing,
suffering from or having a disease, a condition, a disorder, an injury or a
health problem, or a
development or advancement of such states and/or the symptoms of such states.
The treatment or prevention of a disease, a condition, a disorder, an injury
or a health problem may
be partial or complete.
In the formulae of the group which may represent a substituent to the methyl
group in R3, the end point
of the line marked by * in each case does not represent a carbon atom or a CH2
group, but is part of the
bond to the atom to which the group is attached.
In the formulae of the group which may represent R5, the end point of the line
marked by # in each case
does not represent a carbon atom or a CH2 group, but is part of the bond to
the atom to which R5 is
attached.
Preference is given to compounds of the formula (I) in which
represents methyl, ethyl or trifluoromethyl,
R2 represents hydrogen or methyl,
or
RI and R2 together with the carbon atoms to which they are attached form a
cyclobutyl ring,
R3 represents methyl, ethyl or n-propyl,
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where methyl may be substituted with one substituent selected from the group
consisting of
cyclobutyl, tetrahydrofuran-2-yl, tetrahydro-2H-pyran-2-y1 and 1,4-dioxan-2-
yl,
where tetrahydrofuran-2-y1 may be substituted by 1 to 2 substituents methyl,
or
where methyl may be substituted with one substituent of the group of the
formula
R10
R10 OH
0' 0'
*R9 or or
L *R9
where
is the attachment site to the methyl group,
R9 represents methyl, cyclopropyl, difluoromethyl or
trifluoromethyl,
represents methyl or difluoromethyl,
and
where ethyl may be substituted with one substituent selected from the group
consisting of
methoxy, iso-propoxy, tert-butoxy, difluoromethoxy, trifluoromethoxy, 2,2-
difluoroethoxy,
cyclopropyloxy and cyclobutyloxy,
where cyclopropyloxy and cyclobutyloxy may be substituted with one substituent
selected from the group consisting of fluorine and methyl,
R4 represents hydrogen,
R5 represents a group of the formula
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R11
#
0 N H2 or N
l.rN H 2
or #
0 /N¨R12
#
N
0 0
# N # N #
;NI N
..."- --.-
or N¨R13
rx
m15
or
N /( or
le -..... i N-_____
R14
R17
# # #...............NN
N'N N--4
or R16
N N¨C H 3
or or
N Nil
0
0
#
N'I'4
or N¨C H 3
NI
where
# is the attachment site to the nitrogen atom,
RH represents hydrogen or fluorine,
R12 represents difluoromethyl or trifluoromethyl,
RI' represents methyl,
R14 represents hydrogen or methyl,
R15 represents hydrogen or methyl,
R16 represents hydrogen,
R17 represents hydrogen,
R6, R7 and R8 represent the following:
R6 represents hydrogen, fluorine or chlorine,
R7 represents hydrogen,
R8 represents hydrogen,
and the salts thereof, the solvates thereof and the solvates of the salts
thereof
Preference is also given to compounds of the formula (I) in which
RI represents methyl, ethyl or trifluoromethyl,
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R2 represents hydrogen or methyl,
or
RI and R2 together with the carbon atoms to which they are attached form a
cyclobutyl ring,
R3 represents methyl, ethyl or n-propyl,
where methyl may be substituted with one substituent selected from the group
consisting of
cyclobutyl, tetrahydrofuran-2-yl, tetrahydro-2H-pyran-2-y1 and 1,4-dioxan-2-
yl,
where tetrahydrofuran-2-y1 may be substituted by 1 to 2 substituents methyl,
or
where methyl may be substituted with one substituent of the group of the
formula
R10
R10 OH
0' 0'
or or
*LR9 *R9
where
is the attachment site to the methyl group,
R9 represents methyl, cyclopropyl, difluoromethyl or
trifluoromethyl,
represents methyl or difluoromethyl,
and
where ethyl may be substituted with one substituent selected from the group
consisting of
methoxy, iso-propoxy, tert-butoxy, difluoromethoxy, trifluoromethoxy, 2,2-
difluoroethoxy,
cyclopropyloxy and cyclobutyloxy,
where cyclopropyloxy and cyclobutyloxy may be substituted with one substituent
selected from the group consisting of fluorine and methyl,
R4 represents hydrogen,
R5 represents a group of the formula
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NH2 or
H 2
or --.
N R-12
0 0
..======= 15
or N¨R13
or
0
N
¨17Z16
or
or N¨C H 3
Is1/
where
is the attachment site to the nitrogen atom,
RH represents hydrogen or fluorine,
R12 represents difluoromethyl or trifluoromethyl,
represents methyl,
R15 represents hydrogen or methyl,
R16 represents hydrogen,
R6, R7 and R8 represent the following:
R6 represents hydrogen or fluorine,
R7 represents hydrogen,
R8 represents hydrogen,
and the salts thereof, the solvates thereof and the solvates of the salts
thereof
Preference is also given to compounds of the formula (I) in which
RI represents methyl or trifluoromethyl,
R2 represents hydrogen,
or
RI and R2 together with the carbon atoms to which they are attached form a
cyclobutyl ring,
R3 represents methyl or ethyl,
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where methyl is substituted with one substituent selected from the group
consisting of
tetrahydro-2H-pyran-2-y1 and 1,4-dioxan-2-yl,
or
where methyl is substituted with one substituent of the group of the formula
R10
0'
"LR9
where
* is the attachment site to the methyl group,
R9 represents methyl,
Rio represents methyl or difluoromethyl,
and
where ethyl is substituted with one substituent selected from the group
consisting of
methoxy, iso-propoxy, tert-butoxy, difluoromethoxy and cyclopropyloxy,
where cyclopropyloxy may be substituted with one substituent selected from the
group consisting of fluorine and methyl,
R4 represents hydrogen,
R5 represents a group of the formula
# R11
#
0 N H2 or N
N H 2
0 0
# N or or # N
...=- .4....."".....)----....--
N-R13
-R15
N N,N
#
N---N
or )¨ Fil6
N
where
# is the attachment site to the nitrogen atom,
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RH represents hydrogen or fluorine,
represents methyl,
R15 represents hydrogen or methyl,
R16 represents hydrogen,
R6, R7 and R8 represent the following:
R6 represents hydrogen or fluorine,
R7 represents hydrogen,
R8 represents hydrogen,
and the salts thereof, the solvates thereof and the solvates of the salts
thereof
Preference is also given to compounds of the formula (I) in which
represents methyl or trifluoromethyl,
R2 represents hydrogen,
R3 represents methyl,
where methyl is substituted with one substituent of the group of the formula
R10
*) R9
where
is the attachment site to the methyl group,
R9 represents methyl,
Rlo represents methyl or difluoromethyl,
R4 represents hydrogen,
R5 represents a group of the formula
R
N H 2
0
where
is the attachment site to the nitrogen atom,
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RH represents hydrogen,
R6, R7 and R8 represent the following:
R6 represents hydrogen or fluorine,
R7 represents hydrogen,
R8 represents hydrogen,
and the salts thereof, the solvates thereof and the solvates of the salts
thereof
Preference is also given to compounds of the formula (I) in which
R6, R7 and R8 represent the following:
R6 represents hydrogen or fluorine,
R7 represents hydrogen,
R8 represents hydrogen.
Preference is also given to compounds of the formula (I) in which
R6, R7 and R8 represent the following:
R6 represents hydrogen,
R7 represents hydrogen,
R8 represents hydrogen.
Also preferred are compounds having the formula (Ia)
R2
R3
R4
R 0 5
N
0
0
R8
R6
(Ia),
R7
CI
in which RI, R2, R3, R4, R5, R6, R7 and R8 are as defined above.
Preference is also given to the compound 4-(1(2S,45)-2-[(7R)-11-Chloro-2-oxo-7-
(trifluoromethyl)-
2,6,7,8-tetrahydro-3H-[3]benzoxocino[2,1-clpyridin-3-y1]-4-methoxy-
pentanoyl}amino)benzamide
(single stereoisomer) of the formula below
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0C H 3
C H 3
0
N
0 N H 2
0
Ci
or one of the salts thereof, solvates thereof or solvates of the salts thereof
Preference is also given to the compound 4-({(2S,45)-2-[(7R)-11-Chloro-12-
fluoro-2-oxo-7-
(trifluoromethyl)-2,6,7,8-tetrahydro-3H-[3]benzoxocino[2,1-clpyridin-3-yll -4-
methoxy-
pentanoyl}amino)benzamide (single stereoisomer) of the formula below
Cr C H 3
C H 3
0
N
00 N H 2
F 0
CI
or one of the salts thereof, solvates thereof or solvates of the salts thereof
The invention further provides a method for preparing compounds of the formula
(I), or salts thereof,
solvates thereof or solvates of the salts thereof, wherein
[A] the compounds of the formula
R3
R
R12 0 ),.r 0 H
N
0
0
R8
R7
R6
(II),
Ci
in which
RI, R2, R3, R6, R7 and R8 are as defined above,
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are reacted with compounds of the formula
R4
HF,L.5 (III),
in which
R4 and R5 are as defined above,
in the presence of a dehydrating agent to give compounds of the formula (I)
or
[B] the compounds of the formula (II) are converted in a one-pot reaction to
the acid chloride of
compounds of the formula (II) and then the acid chlorides are reacted with
compounds of the formula
(III) to give compounds of the formula (I)
or
[C] the compounds of the formula
R2
R1 0
N H
0
R8
R6
(IV),
R7
CI
in which
RI, R2, R6, R7 and R8 are as defined above,
are reacted with compounds of the formula
R3
R4
X1)-r(V),
0
in which
XI represents bromine, iodine or trifluoromethane-sulfonyloxy,
in the presence of a base to give compounds of the formula (I).
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The reaction according to process [A] is generally carried out in inert
solvents, if appropriate in the
presence of a base, preferably in a temperature range from -20 C to 80 C at
atmospheric pressure.
Alternatively, the reaction can also be carried out without a solvent only in
one base if the base is a
liquid at RT.
Suitable dehydrating agents here are, for example, carbodiimides such as NN'-
diethyl-, NN'-
dipropyl-, NN '-diisopropyl-, NN'-dicyclohexylcarbodiimide, N-(3-
dimethylaminoisopropy1)-N'-
ethylcarbodiimide hydrochloride (EDC) (optionally in the presence of
pentafluorophenol (PFP)), N-
cyclohexylcarbodiimide-N`-propyloxymethyl-polystyrene (PS-carbodiimide) or
carbonyl
compounds such as carbonyldiimidazole, or 1,2-oxazolium compounds such as 2-
ethyl-5-phenyl-
1,2-oxazolium 3-sulfate or 2-tert-butyl-5-methyl-isoxazolium perchlorate, or
acylamino compounds
such as 2-ethoxy-1-ethoxycarbony1-1,2-dihydroquinoline, or isobutyl
chloroformate, or bis-(2-oxo-
3-oxazolidinyl)phosphoryl chloride or benzotriazolyloxytri(dimethylamino)-
phosphonium
hexafluorophosphate, or 0-(benzotriazol-1-y1)-NNN' , N'-tetramethyluronium
hexafluorophosphate
(HBTU), 2 -(2-oxo -1 -(2H)-pyridy1)-1, 1,3,3 -tetramethyluronium
tetrafluoroborate (TPTU),
(benzotriazol-1-yloxy)bisdimethylaminomethylium fluoroborate (TBTU) or 0-(7-
azabenzotriazol-
1-y1)-NNN',Ni-tetramethyluronium hexafluoro-phosphate (HATU), or 1-
hydroxybenzotriazole
(HOBt), or benzotriazol-1-yloxytris(dimethyl-amino)phosphonium
hexafluorophosphate (BOP), or
ethyl cyano(hydroxyimino)acetate (Oxyma), or
(1 -cyano -2-ethoxy-2 -
oxoethylidenaminooxy)dimethylamino -morpholino -carbenium hexafluorophosphate
(COMU), or
N- [(dimethylamino)(3H- [1,2,3] triazolo [4,5 -1)] pyridin-3 -
yloxy)methylidene] -N-
methylmethanaminium hexafluorophosphate, or 2,4,6-tripropy1-1,3,5,2,4,6-
trioxatriphosphinane-
2,4,6-trioxide (T3P), or mixtures of these with bases, the condensation with
HATU or with T3P being
preferred.
Bases are, for example, organic bases such as trialkylamines, for example
triethylamine, N-
methylmorpholine, N-methylpiperidine, 4-dimethylaminopyridine or
diisopropylethylamin, or
pyridine, preference is given to condensation with diisopropylethylamine or
pyridine.
Inert solvents are, for example, halogenated hydrocarbons such as
dichloromethane or
trichloromethane, hydrocarbons such as benzene or toluene, or other solvents
such as 1,4-dioxane,
diethyl ether, tetrahydrofuran, ethyl acetate, N,N-dimethylformamide, dimethyl
sulfoxide or
acetonitrile, or mixtures of the solvents, preference being given to N,N-
dimethylformamide or
tetrahydrofuran.
The reaction according to process [B] is generally carried out in inert
solvents, in the presence of a
chlorination agent, preferably in a temperature range from -20 C to 80 C at
atmospheric pressure.
Inert solvents are, for example, halogenated hydrocarbons such as
dichloromethane or
trichloromethane, hydrocarbons such as benzene or toluene, or other solvents
such as 1,4-dioxane,
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diethyl ether, tetrahydrofuran, ethyl acetate or N,N-dimethylformamide, or
mixtures of the solvents,
preference being given to dichloromethane.
Chlorination agents are, for example, 1-chloro-N,N,2-trimethylprop-1-en- 1 -
amine, oxalyl chloride,
sulfurous dichloride, preference being given to 1-chloro-N,N,2-trimethylprop-1-
en-l-amine.
The reaction according to process [C] is generally carried out in inert
solvents, preferably in a
temperature range from room temperature to reflux of the solvents at
atmospheric pressure.
Bases are, for example, alkali metal hydroxides such as sodium hydroxide,
lithium hydroxide or
potassium hydroxide, or alkali metal carbonates such as caesium carbonate,
sodium carbonate or
potassium carbonate, or potassium tert-butoxide or sodium tert-butoxide,
sodium hydride or a
mixture of these bases or a mixture of sodium hydride and lithium bromide, or
organic bases such as
1,1,3,3 -tetramethylguanidine or 2 -te rt-butylimino -2-diethylamino -1,3 -
dimethylperhydro -1,3,2 -
diazaphosphorine (BEMP), preference is given to potassium carbonate or sodium
hydride or 1,1,3,3-
tetramethylguanidine
Inert solvents are, for example, halogenated hydrocarbons such as
dichloromethane,
trichloromethane, carbon tetrachloride or 1,2-dichloroethane, alcohols such as
methanol, ethanol or
2-propanol, ethers such as diethyl ether, methyl tert-butyl ether, 1,2-
dimethoxyethane, 1,4-dioxane
or tetrahydrofuran, or other solvents such as N,N-dimethylformamide, N,N-
dimethylacetamide,
acetonitrile, pyridine or acetone, or mixtures of solvents, or mixtures of
solvents with water,
preference is given to N,N-dimethylformamide or to a mixture of acetone and 2-
propanol.
The compounds of the formula (III) are known or can be synthesized from the
corresponding starting
compounds by known processes.
The compounds of the formula (V) are known, can be synthesized from the
corresponding starting
compounds by known processes or can be prepared analogously to the processes
described in the
Examples section.
The compounds of the formula (II) are known or can be prepared by reacting
[D] compounds of the formula
R2
R3
R 0
N)-rC)18
0
0
R8
R6
R7
CI
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in which
, R2, R3, R6, R7 and R8 are each as defined above and
R18 represents tert-butyl,
with an acid to give compounds of the formula (II)
or
[E] compounds of the formula
R2
R3
R 0
N 0 R18
0
0
(VIb),
R8
R6
R7
CI
in which
RI, R2, R3, R6, R7 and R8 are each as defined above and
R18 represents methyl, ethyl, tert-butyl or benzyl,
with a base to give compounds of the formula (II).
The reaction according to process [D] is generally carried out in inert
solvents, preferably in a
temperature range from 0 C to 60 C at atmospheric pressure.
Inert solvents are, for example, halogenated hydrocarbons such as
dichloromethane,
trichloromethane, carbon tetrachloride or 1,2-dichloroethane, or ethers such
as tetrahydrofuran or
1,4-dioxane, preference being given to dichloromethane.
Acids are, for example, trifluoroacetic acid or hydrogen chloride in 1,4-
dioxane, preference being
given to trifluoroacetic acid.
The reaction according to process [E] is generally carried out in solvents,
preferably in a temperature
range from room temperature up to reflux of the solvents at atmospheric
pressure.
Inert solvents are, for example, alcohols such as methanol or ethanol, ethers
such as diethyl ether,
methyl tert-butyl ether, 1,2-dimethoxyethane, 1,4-dioxane or tetrahydrofuran,
or mixtures of
solvents, or mixtures of solvents with water, preference being given to a
mixture of tetrahydrofuran
and water.
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Bases are, for example, alkali metal hydroxides such as sodium hydroxide,
lithium hydroxide or
potassium hydroxide, or alkali metal carbonates such as caesium carbonate,
sodium carbonate or
potassium carbonate, preference being given to lithium hydroxide.
The compounds of the formulae (VIa) and (VIb) together form the group of the
compounds of the
formula (VI).
The compounds of the formula (VIa) are known or can be prepared by reacting
compounds of the
formula
R2
R 0
N 0 R18
0
0
R8
R6
R7
CI
in which
RI, R2, R6, R7 and R8 are each as defined above and
R18 represents tert-butyl,
with compounds of the formula
R3¨ X2 (VIII),
in which
R3 is as defined above and
X2 represents chlorine, bromine, iodine or trifluoromethanesulfonyloxy.
The reaction is generally carried out in inert solvents, in the presence of a
base, preferably in a
temperature range from -78 C to room temperature at atmospheric pressure.
Inert solvents are, for example, ethers such as diethyl ether, methyl tert-
butyl ether, 1,2-
dimethoxyethane, 1,4-dioxane or tetrahydrofuran, or mixtures of solvents, or
mixtures of solvent
with water, preference is given to tetrahydrofuran.
Bases are, for example, potassium tert-butoxide or sodium tert-butoxide,
sodium hydride, n-
butyllithium, lithium bis(trimethylsilyl)amide or sodium
bis(trimethylsilyl)amide, preference is
given to lithium bis(trimethylsilyl)amide or sodium bis(trimethylsilyl)amide.
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The compounds of the formula (VIII) are known or can be synthesized from the
corresponding
starting compounds by known processes.
The compounds of the formula (VII) are known or can be prepared by reacting
compounds of the
formula
R2
R 0
N H
0
R8
R6
(IV),
R7
CI
in which
RI, R2, R6, R7 and R8 are as defined above,
with compounds of the formula
x3/ys 8
(IX),
0
in which
X' represents chlorine, bromine, iodine, methanesulfonyloxy or
trifluoromethane-sulfonyloxy
and
R18 represents tert-butyl.
The reaction is carried out as described for process [C].
The compounds of the formula (IX) are known or can be synthesized from the
corresponding starting
compounds by known processes.
In an alternative process, the compounds of the formula (VI) can be prepared
by reacting compounds
of the formula (IV) with compounds of the formula
R3
X
4 -R
18
(X),
0
in which
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R3 is as defined above,
X4 represents chlorine, bromine, iodine, methanesulfonyloxy or
trifluoromethane-sulfonyloxy
and
R18 represents methyl, ethyl, tert-butyl or benzyl.
The reaction is carried out as described for process [C].
The compounds of the formula (X) are known, can be synthesized from the
corresponding starting
compounds by known processes or can be prepared analogously to the processes
described in the
Examples section.
The compounds of the formula (IV) are known or can be prepared by reacting
compounds of the
formula
R2
R1 0
N
C H 3
0
R8
R6
(XI),
R7
Ci
in which
RI, R2, R6, R7 and R8 are as defined above,
with pyridinium hydrochloride or pyridinium hydrobromide or sodium iodide and
an acid or lithium
iodide and an acid or with a thiol, such as ethanethiol, and a base.
The reaction is generally carried out in inert solvents or without solvents,
preferably in a temperature
range of from 80 C to 120 C at atmospheric pressure.
Inert solvents are, for example, hydrocarbons such as benzene, or alcohols
such as methanol, ethanol
or 1-butanol, or other solvents such as nitromethane, 1,4-dioxane, N,N-
dimethylformamide, dimethyl
sulfoxide or acetonitrile, or a mixture of the solvents, preference is given
to N,N-dimethylformamide
or 1-butanol.
Acids are, for example, 4-toluenesulfonic acid monohydrate, formic acid,
acetic acid, trifluoroacetic
acid, preference is given to acetic acid and 4-toluenesulfonic acid
monohydrate.
Bases are, for example, potassium tert-butoxide or sodium tert-butoxide or
sodium hydride,
preference is given to sodium hydride.
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The compounds of the formula (XI) are known or can be prepared by reacting
compounds of the
formula
R7
CI R8
0
R6
N (XII),
R2 I0'CH3
in which
RI, R2, R6, R7 and R8 are as defined above,
with a base in the presence of a catalyst.
The reaction is generally carried out in inert and degassed solvents,
preferably within a temperature
range from 80 C to 150 C at atmospheric pressure.
Bases are, for example, alkali metal carbonates such as sodium carbonate or
potassium carbonate,
organic bases such as sodium pivalate, potassium pivalate, caesium pivalate,
sodium acetate,
potassium acetate, caesium acetate, preference is given to potassium pivalate
or potassium acetate.
Catalysts are, for example, palladium catalysts customary for CH-activation
conditions, such as
dichlorobis(triphenylphosphine)palladium,
tetrakis(triphenylphosphine)palladium(0), palladium(II)
acetate/triscyclohexylphosphine, bis(tri-tert-butylphosphine)palladium(0),
tris(dibenzylidene-
acetone)dipalladium, bis(diphenylphosphaneferrocenyl)palladium(II) chloride,
1,3-bis(2,6-
diisopropylphenypimidazol-2-ylidene(1,4-naphthoquinone)palladium dime r, allyl
(chloro) (1,3 -
dime sityl -1,3 -dihydro -2H-imidazol-2-ylidene)palladium,
palladium(II) acetate/
dicyclohexyl(2',4',6'-triisopropyl-biphenyl-2-yl)phosphine, XPhos precatalyst
[(2'-aminobipheny1-2-
yl)(chloro)palladium dicyclohexyl(2',4',6'-triisopropylbipheny1-2-yl)phosphane
(1:1)1, DavePhos
precatalyst [Methane sulfonato 2-dicyclohexylphosphino-2-(N,N-
dimethylamino)bipheny1(21-amino-
1,11-bipheny1-2-y1) palladium(II)1, PEPPSIO-catalysts
such as [1,3-bis(2,6-di-3-
pentylphenyl)imidazol-2-ylidenel (3 -chloropyridyl)palladium(II) dichloride
or [1,3 -bi s (2,6-
diisopropylphenypimidazol-2-ylidenel (3-chloropyridyl)palladium(II)
dichloride, preference being
given to tetrakis(triphenylphosphine)palladium(0) or [1,3-bis(2,6-di-3-
pentylphenyl)imidazol-2-
ylidene] (3 -chloropyridyl)palladium(II) dichloride.
Inert solvents are, for example, ethers such as 1,4-dioxane, tetrahydrofuran
or 1,2-dimethoxyethane,
hydrocarbons such as benzene, xylene or toluene, or carboxamides such as N,N-
dimethylformamide
or N,N-dimethylacetamide, alkyl sulfoxides such as dimethyl sulfoxide, or
mixtures of the solvents
with water, preference is given to N,N-dimethylformamide or N,N-
dimethylacetamide.
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In an alternative process, the compounds of the formula (XI) can be prepared
by reacting compounds
of the formula
R7
C I R8
0
R6
N (XIII),
Br R2
H 3
0
in which
RI, R2, R6, R7 and R8 are as defined above,
with bis(pinacolato)diboran in the presence of a base and a catalyst.
The reaction is generally carried out in inert solvents, preferably within a
temperature range from
room temperature to 150 C at atmospheric pressure.
Catalysts are, for example, palladium catalysts customary for Suzuki reaction
conditions, preference
being given to catalysts such as dichlorobis(triphenylphosphine)palladium,
tetrakistriphenylphosphinepalladium(0), bis(tri-tert-
butylphosphine)palladium(0), palladium(II)
acetate, palladium(II) acetate/triscyclohexylphosphine,
tris(dibenzylideneacetone)dipalladium,
bis(diphenylphosphaneferrocenyl)palladium(II) chloride, 1,3 -bi s (2,6-dii
sopropylphenypimidazol -2 -
ylidene (1,4-naphthoquinone)palladium
dimer, allyl (chloro)(1,3 -dime sityl-1,3 -dihydro -2H-
imidazol-2-ylidene)palladium, palladium(II) acetate/ dicyclohexyl(2',4',6'-
triisopropyl-bipheny1-2-
yl)phosphine, [1,1-bis(diphenylphosphino)-ferrocene]palladium(II) chloride
monodichloromethane
adduct or XPhos precatalyst [(2'-aminobipheny1-2-y1)(chloro)palladium
dicyclohexyl(2',4',6'-
triisopropylbipheny1-2-yl)phosphane (1:1)1, preference being given to
palladium(II) acetate or
palladium(II) acetate/triscyclohexylphosphine or
tetrakistriphenylphosphinepalladium(0).
Bases are, for example, alkali metal carbonates such as caesium carbonate,
potassium carbonate or
sodium carbonate, alkali metal bicarbonates such as, sodium bicarbonate,
alkali or alkaline earth
metal hydroxides such as sodium hydroxide, potassium hydroxide, barium
hydroxide or potassium
acetate, potassium tert-butoxide or potassium phosphate, where these may be
present in aqueous
solution, preference being given to an aqueous sodium bicarbonate solution or
potassium acetate.
Inert solvents are, for example, ethers such as 1,4-dioxane, tetrahydrofuran
or 1,2-dimethoxyethane,
hydrocarbons such as benzene, xylene or toluene, or carboxamides such as N,N-
dimethylformamide
or N,N-dimethylacetamide, alkyl sulfoxides such as dimethyl sulfoxide, or N-
methylpyrrolidone or
acetonitrile, or mixtures of the solvents with alcohols such as methanol or
ethanol and/or water,
preference is given to N,N-dimethylformamide.
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The compounds of the formula (XII) are known or can be prepared by reacting
compounds of the
formula
R7
C I R8 1
R6 0 H (XIV)'
R2
in which
RI, R2, R6, R7 and R8 are as defined above,
with a compound of the formula
H 0
C H 3 (XV)
I C)
The reaction is carried out in the presence of (tributylphosphoranylidene)-
acetonitrile, in inert
solvents such as benzene, xylene or toluene, preference is given to toluene,
optionally in a
microwave, preferably within a temperature range from 80 C to 160 C at
atmospheric pressure to 3
bar or higher than 3 bar using a microwave,
or
the reaction is carried out in the presence of an ester of the azodicarboxylic
acid such as diisopropyl
azodicarboxylate (DIAD) and organophosphorus compounds such as
triphenylphosphine, in inert
solvents such as tetrahydrofuran, 1,4-dioxane or dichloromethane, within a
temperature range of 0 C
to room temperature at atmospheric pressure.
The compounds of the formulae (XIV) and (XV) are known, can be synthesized
from the
corresponding starting compounds by known processes or can be prepared
analogously to the
processes described in the Examples section.
The compounds of the formula (XIII) are known or can be prepared by reacting
compounds of the
formula
R7
C I R8
0 H (XVI),
R6
Br R2
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in which
RI, R2, 12_6, R7 and R8 are as defined above,
with a compound of the formula (XV).
The reaction is carried out as described for the reaction of compounds of the
formula (XIV) with
compounds of the formula (XV) .
The compounds of the formula (XVI) are known, can be synthesized from the
corresponding starting
compounds by known processes or can be prepared analogously to the processes
described in the
Examples section.
The preparation of the starting compounds and of the compounds of the formula
(I) can be illustrated
by the synthesis scheme which follows.
Scheme:
HO
R7
CI ReRI I -"......**-"J'...
. lC H3 R7
CI R8
R6 1
R
0 H ______________________ ..
R2
R6 0 N
Br R2
R1 0
Br R2 .....õ-Ø.... C F3". ...,- N
CH.,
0'
R7 HO
R8 R1 .../....¶N Re R6
'
R7
CI 'C H3 CI R8 1
7 R7 CI
R
Re
R20 H
R6 R2 -.... I Orj
'C H 3
I
I 0
R2 R2
0 0 C H3 0
Ri õ.õ.. N.........y,0,,,,C H3 )(3r )< CH3
R1 ...".. N H
r-O H3 0 C H 3
H 3
0 0
R8 R6 0 C
Re
R8
R7 CI R3
H3 R7 CI X X4
....,....õC H3
0 C H3 R3 R4
R3¨X2 i I
l)yN 5
R
0
R2 R3
,
R2 R3
0 R2 R3 R4
0 ....,, 0 C H
R1 ,,,3 l
R ),,,r,0 H I
...," N ...," N R4 0
r- c H3 I Ri .."'"
elyN'sR6
R8 Re
0 0 C H3 R8 0 H N
0 ***'126 _,..
R
Re ________________________________________________________ .. R8 0
0
6
R7 CI R7 CI
R7 CI
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The compounds according to the invention have an unforeseeable useful
pharmacological activity
spectrum and good pharmacokinetic properties. They are compounds that
influence the proteolytic
activity of the serine protease factor XIa (FXIa). The compounds according to
the invention inhibit
the enzymatic cleavage of FXIa-substrates, such as factor IX (FIX), which have
essential roles in the
activation of blood coagulation, in the aggregation of blood platelets via PAR-
1 activation of the
platelets, and in inflammatory processes, which particularly involve an
increase in vascular
permeability.
They are therefore suitable for use as medicaments for the treatment and/or
prophylaxis of diseases
in humans and animals.
The present invention further provides for the use of the compounds according
to the invention for
the treatment and/or prophylaxis of disorders, in particular vascular
disorders, preferably thrombotic
or thromboembolic disorders and/or thrombotic or thromboembolic complications.
Factor XIa (FXIa) is an important enzyme in the context of coagulation, which
can be activated by
both thrombin and factor XIIa (FXIIa), and is therefore involved in two
essential processes of
coagulation. It is a central component of the transition from initiation to
amplification of the
coagulation and propagation of the clot: in positive feedback loops, thrombin
activates, in addition
to factor V and factor VIII, also factor XI to factor XIa, whereby factor IX
is converted into factor
IXa, and, via the factor IXa/factor Villa complex generated in this manner,
factor Xa and
subsequently thrombin are formed, leading to strong thrombus growth and
stabilization of the
thrombus.
Moreover, factor XIa is an important component for the intrinsic initiation of
coagulation: In addition
to the stimulation via tissue factor (TF) in the extrinsic pathway, the
coagulation system can be
activated also particularly on negatively charged surfaces, which include not
only surface structures
of foreign cells (e.g. bacteria) but also artificial surfaces such as vascular
prostheses, stents and parts
of extracorporeal circulation systems. On these surfaces, factor XII (FXII) is
activated to factor XIIa
(FXIIa) which subsequently activates FXI to FXIa. This leads to further
activation of the coagulation
cascade as described above.
In contrast, thrombin generation triggered by TF/factor VIIa via factor X
activation and finally
thrombin formation, which represents the early physiological reaction to
vascular wall injuries,
remains uninfluenced. This could explain why no prolongations of bleeding
times were found in
FXIa knockout mice, as in rabbits and other species, with administration of
FXIa inhibitor. This low
bleeding tendency caused by the substance is of great advantage for use in
humans, particularly in
patients with increased risk of bleeding.
Accordingly, the compounds according to the invention are suitable for the
treatment and/or
prophylaxis of disorders or complications which may arise from the formation
of clots.
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For the purpose of the present invention, the "thrombotic or thromboembolic
disorders and/or
thrombotic or thromboembolic complications" include disorders and
complications, which occur in
the arterial, the venous vascular system and the lymphatic system, which can
be treated with the
compounds according to the invention. This includes in particular disorders in
the coronary arteries
of the heart, such as acute coronary syndrome (ACS), myocardial infarction
with ST segment
elevation (STEMI) and without ST segment elevation (non-STEMI), stable angina
pectoris, unstable
angina pectoris, stent thrombosis, reocclusions and restenoses after coronary
interventions such as
angioplasty, stent implantation or aortocoronary bypass, disorders in the
cerebrovascular arteries,
such as transitory ischaemic attacks (TIA), ischemic strokes including
cardioembolic strokes, such
as strokes due to atrial fibrillation, non-cardioembolic strokes, such as
lacunar stroke, strokes due to
large or small artery diseases, or strokes due to undetermined cause,
cryptogenic strokes, embolic
strokes, embolic strokes of undetermined source, or events of thrombotic
and/or thromboembolic
origin leading to stroke or TIA, and disorders of peripheral arteries, leading
to peripheral artery
disease, including peripheral artery occlusion, acute limb ischemia,
amputation, reocclusions and
restenoses after interventions such as angioplasty, stent implantation or
surgery and bypass.
In addition, this includes thrombotic or thromboembolic disorders in
particular in veins of the
extremities, kidneys, mesenterium, liver, brain and eye, leading to pulmonary
embolisms, venous
thromboembolisms and/or venous thrombosis.
Stimulation of the coagulation system may occur by various causes or
associated disorders. In the
context of surgical interventions, immobility, confinement to bed, infections,
inflammation or cancer
or cancer therapy, inter alia, the coagulation system can be highly activated,
and there may be
thrombotic complications, in particular venous thromboses. The compounds
according to the
invention are therefore suitable for the prophylaxis of thrombosis in the
context of surgical
interventions in patients suffering from cancer. The compounds according to
the invention are
therefore also suitable for the prophylaxis of thrombosis in patients having
an activated coagulation
system, for example in the situations described above.
The inventive compounds are therefore also suitable for the prevention and
treatment of cardiogenic
thromboembolisms, for example brain ischaemias, stroke and systemic
thromboembolisms and
ischaemias, in patients with acute, intermittent or persistent cardiac
arrhythmias, for example atrial
fibrillation, and in patients undergoing cardioversion, and also in patients
with heart valve disorders
or with artificial heart valves.
In addition, the inventive compounds are suitable for the treatment and
prevention of disseminated
intravascular coagulation (DIC) which may occur in connection with sepsis
inter alia, but also owing
to surgical interventions, neoplastic disorders, burns or other injuries and
may lead to severe organ
damage through microthrombosis.
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Thromboembolic complications furthermore occur in microangiopathic
haemolytical anaemias and
by blood coming into contact with artificial surfaces in the context of
extracorporeal circulation such
as, for example, haemodialysis and ECM ("extracorporeal membrane
oxygenation"), LVAD ("left
ventricular assist device") and similar devices, AV fistulas, vascular and
heart valve prostheses.
Moreover, the compounds according to the invention are suitable for the
treatment and/or
prophylaxis of disorders involving microclot formation or fibrin deposits in
cerebral blood vessels
or asymptomatic, covert strokes, which may lead to dementia disorders such as
vascular dementia or
Alzheimer's disease. Here, the clot may contribute to the disorder both via
occlusions and by binding
disease-relevant factors.
Moreover, the compounds according to the invention are suitable for the
treatment and/or
prophylaxis of disorders where, in addition to the pro-coagulant component,
the pro-inflammatory
component also plays an essential role. Mutual enhancement of coagulation and
inflammation in
particular can be prevented by the compounds according to the invention, thus
decisively lowering
the probability of thrombotic complications. Therefore, the compounds
according to the invention
are suitable for the treatment and/or prophylaxis in the context of
atherosclerotic vascular disorders,
inflammatory diseases, such as rheumatic disorders of the locomotor system,
inflammatory disorders
of the lung, such as pulmonary fibroses, inflammatory disorders of the kidney,
such as
glomerulonephritides, inflammatory disorders of the intestine, such as Crohn's
disease or ulcerative
colitis, or disorders, which may be present in the context of an underlying
diabetic disease, such as
diabetic retinopathy or nephropathy.
Moreover, the compounds according to the invention can be used for inhibiting
tumor growth and
the formation of metastases, and also for the prophylaxis and/or treatment of
thromboembolic
complications, such as, for example, venous thromboembolisms, for cancer
patients, in particular
those undergoing major surgical interventions or chemo- or radiotherapy.
In addition, the inventive compounds are also suitable for the prophylaxis
and/or treatment of
pulmonary hypertension.
In the context of the present invention, the term "pulmonary hypertension"
includes pulmonary
arterial hypertension, pulmonary hypertension associated with disorders of the
left heart, pulmonary
hypertension associated with pulmonary disorders and/or hypoxia and pulmonary
hypertension
owing to chronic thromboembolisms (CTEPH).
"Pulmonary arterial hypertension" includes idiopathic pulmonary arterial
hypertension (IPAH,
formerly also referred to as primary pulmonary hypertension), familial
pulmonary arterial
hypertension (FPAH) and associated pulmonary arterial hypertension (APAH),
which is associated
with collagenoses, congenital systemic-pulmonary shunt vitia, portal
hypertension, HIV infections,
the ingestion of certain drugs and medicaments, with other disorders (thyroid
disorders, glycogen
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storage disorders, Morbus Gauche r, hereditary teleangiectasia,
haemoglobinopathie s,
myeloproliferative disorders, splenectomy), with disorders having a
significant venous/capillary
contribution, such as pulmonary-venoocclusive disorder and pulmonary-capillary
haemangiomatosis, and also persisting pulmonary hypertension of neonatants.
.. Pulmonary hypertension associated with disorders of the left heart includes
a diseased left atrium or
ventricle and mitral or aorta valve defects.
Pulmonary hypertension owing to chronic thromboembolisms (CTEPH) comprises the
thromboembolic occlusion of proximal pulmonary arteries, the thromboembolic
occlusion of distal
pulmonary arteries and non-thrombotic pulmonary embolisms (tumour, parasites,
foreign bodies).
.. The present invention further provides for the use of the inventive
compounds for production of
medicaments for the treatment and/or prophylaxis of pulmonary hypertension
associated with
sarcoidosis, histiocytosis X and lymphangiomatosis.
In addition, the compounds according to the invention may also be useful for
the treatment of lung,
liver and kidney fibrosis.
The compounds according to the invention are also suitable for the primary
prophylaxis of
thrombotic or thromboembolic disorders and/or thrombo-inflammatory disorders
and/or disorders
with increased vascular permeability in patients, in which gene mutations lead
to enhanced activity
of the enzymes or increased levels of the zymogens - and these are established
by relevant
tests/measurements of the enzyme activity or zymogen concentrations.
The present invention further provides for the use of the compounds according
to the invention for
the treatment and/or prophylaxis of disorders, especially the disorders
mentioned above.
The present invention further provides for the use of the compounds according
to the invention for
production of a medicament for the treatment and/or prophylaxis of disorders,
especially the
disorders mentioned above.
The present invention further provides a method for the treatment and/or
prophylaxis of disorders,
especially the disorders mentioned above, using a therapeutically effective
amount of a compound
according to the invention.
The present invention further provides the compounds according to the
invention for use in a method
for the treatment and/or prophylaxis of disorders, especially the disorders
mentioned above, using a
therapeutically effective amount of a compound according to the invention.
The present invention further provides medicaments comprising a compound
according to the
invention and one or more further active compounds.
In addition, the compounds according to the invention can also be used for
preventing coagulation
ex vivo, for example for the protection of organ transplants against damage
caused by formation of
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clots and for protecting the organ recipient against thromboemboli from the
transplanted organ, for
preserving blood and plasma products, for cleaning/pretreating catheters and
other medical
auxiliaries and instruments, for coating synthetic surfaces of medical
auxiliaries and instruments used
in vivo or ex vivo or for biological samples which may contain factor XIa.
The present invention furthermore provides a method for preventing the
coagulation of blood in vitro,
in particular in banked blood or biological samples which may comprise factor
XIa, which method
is characterized in that an anticoagulatory effective amount of the compound
according to the
invention is added.
The present invention further provides medicaments comprising a compound
according to the
invention and one or more further active compounds, in particular for the
treatment and/or
prophylaxis of the disorders mentioned above. Preferred examples of active
compounds suitable for
combinations include:
= lipid-lowering substances, especially HMG-CoA (3-hydroxy-3-methylglutaryl-
coenzyme A)
reductase inhibitors, for example lovastatin (Mevacor), simvastatin (Zocor),
pravastatin
(Pravachol), fluvastatin (Lescol) and atorvastatin (Lipitor);
= coronary therapeutics/vasodilators, especially ACE (angiotensin
converting enzyme) inhibitors,
for example captopril, lisinopril, enalapril, ramipril, cilazapril,
benazepril, fosinopril, quinapril
and perindopril, or All (angiotensin II) receptor antagonists, for example
embusartan, losartan,
valsartan, irbesartan, candesartan, eprosartan and temisartan, or fl-
adrenoceptor antagonists, for
example carvedilol, alprenolol, bisoprolol, acebutolol, atenolol, betaxolol,
carteolol, metoprolol,
nadolol, penbutolol, pindolol, propanolol and timolol, or alpha- 1 -
adrenoceptor antagonists, for
example prazosine, bunazosine, doxazosine and terazosine, or diuretics, for
example
hydrochlorothiazide, furosemide, bumetanide, piretanide, torasemide, amiloride
and
dihydralazine, or calcium channel blockers, for example verapamil and
diltiazem, or
dihydropyridine derivatives, for example nifedipin (Adalat) and nitrendipine
(Bayotensin), or
nitro preparations, for example isosorbide 5 -mononitrate, isosorbide
dinitrate and glycerol
trinitrate, or substances causing an increase in cyclic guanosine
monophosphate (cGMP), for
example stimulators of soluble guanylate cyclase, for example riociguat;
= plasminogen activators (thrombolytics/fibrinolytics) and compounds which
promote
thrombolysis/fibrinolysis such as inhibitors of the plasminogen activator
inhibitor (PAT
inhibitors) or inhibitors of the thrombin-activated fibrinolysis inhibitor
(TAFI inhibitors) such
as, for example, tissue plasminogen activator (t-PA, for example Actilyse),
streptokinase,
reteplase and urokinase or plasminogen-modulating substances causing increased
formation of
plasmin;
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= anticoagulatory substances (anticoagulants), for example heparin (UFH),
low-molecular-weight
heparins (LMW), for example tinzaparin, certoparin, parnaparin, nadroparin,
ardeparin,
enoxaparin, reviparin, dalteparin, danaparoid, semuloparin (AVE 5026),
adomiparin (M118)
and EP-42675/0RG42675;
= direct thrombin inhibitors (DTI) such as, for example, Pradaxa
(dabigatran), atecegatran (AZD-
0837), DP-4088, SSR-182289A, argatroban, bivalirudin and tanogitran (BIBT-986
and prodrug
BIBT-1011), hirudin;
= direct factor Xa inhibitors, for example, rivaroxaban, apixaban, edoxaban
(DU-176b),
betrixaban (PRT-54021), R-1663, darexaban (YM-150), otamixaban (FXV-673/RPR-
130673),
letaxaban (TAK-442), razaxaban (DPC-906), DX-9065a, LY-517717, tanogitran
(BIBT-986,
prodrug: BIBT-1011), idraparinux and fondaparinux,
= substances which inhibit the aggregation of platelets (platelet
aggregation inhibitors,
thrombocyte aggregation inhibitors), such as, for example, acetylsalicylic
acid (such as, for
example, aspirin), P2Y12 antagonists such as, for example, ticlopidine
(Ticlid), clopidogrel
(Plavix), prasugrel, ticagrelor, cangrelor, elinogrel, PAR-1 antagonists such
as, for example,
vorapaxar, PAR-4 antagonists, EP3 antagonists such as, for example, DG041;
= platelet adhesion inhibitors such as GPVI and/or GPIb antagonists such
as, for example,
Revacept or caplacizumab;
= fibrinogen receptor antagonists (glycoprotein-IIWIlla antagonists), for
example abciximab,
eptifibatide, tirofiban, lamifiban, lefradafiban and fradafiban;
= recombinant human activated protein C such as, for example, Xigris or
recombinant
thrombomudulin;
= and also antiarrhythmics.
"Combinations" for the purpose of the invention mean not only dosage forms
which contain all the
components (so-called fixed combinations) and combination packs which contain
the components
separate from one another, but also components which are administered
simultaneously or
sequentially, provided that they are used for prophylaxis and/or treatment of
the same disease. It is
likewise possible to combine two or more active ingredients with one another,
meaning that they are
thus each in two-component or multicomponent combinations.
The inventive compounds can act systemically and/or locally. For this purpose,
they can be
administered in a suitable manner, for example by the oral, parenteral,
pulmonal, nasal, sublingual,
lingual, buccal, rectal, dermal, transdermal, conjunctival or otic route, or
as an implant or stent.
The inventive compounds can be administered in administration forms suitable
for these
administration routes.
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Suitable administration forms for oral administration are those which function
according to the prior
art and deliver the inventive compounds rapidly and/or in modified fashion,
and which contain the
inventive compounds in crystalline and/or amorphized and/or dissolved form,
for example tablets
(uncoated or coated tablets, for example having enteric coatings or coatings
which are insoluble or
dissolve with a delay, which control the release of the compound according to
the invention), tablets
which disintegrate rapidly in the mouth, or films/wafers, films/lyophilisates,
capsules (for example
hard or soft gelatin capsules), sugar-coated tablets, granules, pellets,
powders, emulsions,
suspensions, aerosols or solutions.
Parenteral administration can be accomplished with avoidance of a resorption
step (for example by
an intravenous, intraarterial, intracardiac, intraspinal or intralumbar route)
or with inclusion of a
resorption (for example by an intramuscular, subcutaneous, intracutaneous,
percutaneous or
intraperitoneal route). Administration forms suitable for parenteral
administration include
preparations for injection and infusion in the form of solutions, suspensions,
emulsions, lyophilizates
or sterile powders.
Preference is given to oral administration.
Suitable administration forms for the other administration routes are, for
example, pharmaceutical
forms for inhalation (including powder inhalers, nebulizers), nasal drops,
solutions or sprays; tablets
for lingual, sublingual or buccal administration, films/wafers or capsules,
suppositories, preparations
for the ears or eyes, vaginal capsules, aqueous suspensions (lotions, shaking
mixtures), lipophilic
suspensions, ointments, creams, transdermal therapeutic systems (for example
patches), milk, pastes,
foams, dusting powders, implants or stents.
The inventive compounds can be converted to the administration forms
mentioned. This can be
accomplished in a manner known per se by mixing with inert, nontoxic,
pharmaceutically suitable
excipients. These excipients include carriers (for example microcrystalline
cellulose, lactose,
mannitol), solvents (e.g. liquid polyethylene glycols), emulsifiers and
dispersing or wetting agents
(for example sodium dodecylsulfate, polyoxysorbitan oleate), binders (for
example
polyvinylpyrrolidone), synthetic and natural polymers (for example albumin),
stabilizers (e.g.
antioxidants, for example ascorbic acid), colourants (e.g. inorganic pigments,
for example iron
oxides) and flavour and/or odour correctants.
The present invention further provides medicaments comprising at least one
inventive compound,
preferably together with one or more inert nontoxic pharmaceutically suitable
excipients, and the use
thereof for the purposes mentioned above.
In the case of parenteral administration, it has generally been found to be
advantageous to administer
amounts of about 5 to 250 mg every 24 hours to achieve effective results. In
the case of oral
administration, the amount is about 5 to 500 mg every 24 hours.
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In spite of this, it may be necessary, if appropriate, to deviate from the
amounts specified, specifically
depending on body weight, administration route, individual behaviour towards
the active ingredient,
type of formulation, and time or interval of administration.
Unless stated otherwise, the percentages in the tests and examples which
follow are percentages by
weight; parts are parts by weight. Solvent ratios, dilution ratios and
concentration data for the
liquid/liquid solutions are based in each case on volume. "w/v" means
"weight/volume". For
example, "10% w/v" means: 100 ml of solution or suspension comprise 10 g of
substance.
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A) Examples
Abbreviations:
Boc tert-Butyloxycarbonyl
br s broad singlet (in NMR)
br d broad doublet (in NMR)
br t broad triplet (in NMR)
cv column volume
d day(s), doublet (in NMR)
TLC thin-layer chromatography
DCI direct chemical ionization (in MS)
dd doublet of doublets (in NMR)
ddd doublet of doublets of doublets (in NMR)
DMSO dimethyl sulfoxide
eq. equivalent(s)
ESI electrospray ionization (in MS)
h hour(s)
HATU 0-(7-azabenzotriazol-1-y1)-/V,/V,M,M-tetramethyluronium
hexafluorophosphate
HPLC high-pressure, high-performance liquid chromatography
LC/MS liquid chromatography-coupled mass spectroscopy
m multiplet (in NMR)
min minute(s)
MS mass spectroscopy
NMR nuclear magnetic resonance spectroscopy
q quartet or quadruplet (in NMR)
quin quintet (in NMR)
RP reverse phase (in HPLC)
RT room temperature
Rt retention time (in HPLC)
s singlet (in NMR)
sxt sextet (in NMR)
SFC supercritical fluid chromatography (with supercritical
carbon dioxide as a
eluent)
t triplet (in NMR)
TFA trifluoroacetic acid
T3P 2,4,6-tripropy1-1,3,5,2,4,6-trioxatriphosphinane 2,4,6-
trioxide
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HPLC, LC-MS and GC methods:
Method 1: Instrument: Waters ACQUITY SQD UPLC system; column: Waters Acquity
UPLC HSS
T3 C18 1.8 [tm, 50 mm x 1.0 mm; eluent A: water + 0.025% formic acid, eluent
B: acetonitrile +
0.025% formic acid; gradient: 0.0 min 10% B -> 1.2 min 95% B -> 2.0 min 95% B;
oven: 50 C;
flow rate: 0.40 ml/min; UV detection: 210-400 nm.
Method 2: Instrument: Thermo Scientific DSQII; GC: Thermo Scientific Trace GC
Ultra; column:
Restek RTX-35M5, 15 m x 200 [tm x 0.33 [tm; constant helium flow rate: 1.20
ml/min; oven: 60 C;
inlet: 220 C; gradient: 60 C, 30 C/min -> 300 C (maintained for 3.33 min).
Method 3: Instrument: Waters ACQUITY SQD UPLC system; column: Waters Acquity
UPLC HSS
T3 C18 1.8 [tm, 50 mm x 1.0 mm; eluent A: water + 0.025% formic acid, eluent
B: acetonitrile +
0.025% formic acid; gradient: 0.0 min 5% B -> 6.0 min 95% B -> 7.5 min 95% B;
oven: 50 C; flow
rate: 0.35 ml/min; UV detection: 210-400 nm.
Method 4: Instrument: Thermo Scientific FT-MS; UHPLC: Thermo Scientific
UltiMate 3000;
column: Waters HSS T3 C18 1.8 [tm, 75 mm x 2.1 mm; eluent A: water + 0.01%
formic acid; eluent
.. B: acetonitrile + 0.01% formic acid; gradient: 0.0 min 10% B -> 2.5 min 95%
B -> 3.5 min 95% B;
oven: 50 C; flow rate: 0.90 ml/min; UV detection: 210-400 nm.
Method 5: Instrument: Agilent MS Quad 6150; HPLC: Agilent 1290; column: Waters
Acquity UPLC
HSS T3 1.8 [tm, 50 mm x 2.1 mm; eluent A: water + 0.025% formic acid, eluent
B: acetonitrile +
0.025% formic acid; gradient: 0.0 min 10% B -> 0.3 min 10% B -> 1.7 min 95% B -
> 3.0 min 95%
B; oven: 50 C; flow rate: 1.20 ml/min; UV detection: 205-305 nm.
Method 6: Instrument: Waters MS SQ detector 2; GC: Agilent A7890; column:
Restek RTX-35 MS,
15 m x 200 [tm x 0.33 [tm, gas: helium; oven: 60 C; flow rate: 1.20 ml/min;
inlet: 240 C; gradient:
C/min -> 300 C.
Method 7: Instrument: Shimadzu LCMS-2020; column: CORTECS C18 2.7 [tm, 50 mm x
2.1 mm;
25 eluent A: water + 0.1% formic acid, eluent B: acetonitrile + 0.1% formic
acid, gradient: 0.0 min 5%
B -> 2.0 min 95% B -> 3.0 min 95% B; oven: 40 C; flow rate: 1.0 ml/min; UV
detection: 210-400
nm.
Method 8: Instrument: Shimadzu LCMS-2020; column: CORTECS C18 2.7 [tm, 2.1 mm
x 50 mm;
eluent A: water + 0.1% formic acid, eluent B: acetonitrile + 0.1% formic acid,
gradient: 0.0 min 5%
30 B -> 1.2 min 100% B -> 2.0 min 100% B; oven: 40 C; flow rate: 1.0
ml/min; UV detection: 210-
400 nm.
Method 9: Instrument: Shimadzu LCMS-2020; column: Kinetex EVO-C18 2.6 [tm, 3.0
mm x
50 mm; eluent A: water + 0.2% ammonium hydroxide, eluent B: acetonitrile,
gradient: 0.0 min 10%
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B -> 2.0 min 95% B -> 3.0 min 95% B; oven: 45 C; flow rate: 1.2 ml/min; UV
detection: 210-400
nm.
Method 10: Instrument: Shimadzu LCMS-2020; column: CORTECS C18 2.7 pm, 50 mm x
2.1 mm;
eluent A: water + 0.09% formic acid, eluent B: acetonitrile + 0.1% formic
acid, gradient: 0.0 min 5%
B -> 1.2 min 100% B -> 2.0 min 100% B; oven: 40 C; flow rate: 1.0 ml/min; UV
detection: 210-
400 nm.
Method 11: Instrument: Shimadzu LC-MS-2020; column: Ascentis Express C18 2.7
[tm, 50 mm x
2.1 mm; eluent A: water with 0.05% trifluoroacetic acid, eluent B:
acetonitrile with 0.05%
trifluoroacetic acid; gradient: 0.0 min 5% B -> 1.2 min 100% B -> 1.7 min 100%
B -> 1.75 min 5%
B -> 2.0 min 5% B; oven: 40 C; flow rate: 1.5 ml/min; UV detection: 210-400
nm.
Method 12: Instrument: Shimadzu LC-MS-2020; column: Ascentis Express C18 2.7
[tm, 50 mm x
3.0 mm; eluent A: water with 0.05% trifluoroacetic acid, eluent B:
acetonitrile with 0.05%
trifluoroacetic acid; gradient: 0.0 min 5% B -> 1.2 min 95% B -> 1.7 min 95% B
-> 1.8 min 5% B
-> 2.0 min 5% B; oven: 40 C; flow rate: 1.5 ml/min; UV detection: 210-400 nm.
Method 13: Instrument: Shimadzu LCMS-2020; column: Kinetex EVO C18 2.6 [tm, 50
mm x
3.0 mm; eluent A: water + 0.03% ammonium hydroxide, eluent B: acetonitrile,
gradient: 0.0 min
10% B -> 1.1 min 95% B -*2.0 min 95% B; oven: 40 C; flow rate: 1.2 ml/min; UV
detection: 210-
400 nm.
Method 14: Instrument: Shimadzu LCMS-2020; column: Kinetex EVO C18 2.6 [tm, 50
mm x
3.0 mm; eluent A: water + 0.03% ammonium hydroxide, eluent B: acetonitrile,
gradient: 0.0 min
10% B -> 2.0 min 95% B -> 3.0 min 95% B; oven: 40 C; flow rate: 1.2 ml/min; UV
detection: 210-
400 nm.
Method 15: Instrument: Waters Single Quad MS; HPLC: Waters UPLC Acquity;
column: Waters
BEH C18 1.7 [tm, 50 mm x 2.1 mm; eluent A: water + 0.025% ammonia, eluent B:
acetonitrile;
gradient: 0.0 min 8% B -> 0.1 min 8% B -> 1.8 min 95% B -> 3.5 min 95% B;
oven: 50 C; flow
rate: 0.45 ml/min; UV detection: 210-400 nm.
Method 16: Instrument: Waters TOF instrument; UPLC: Waters Acquity I-CLASS;
column: Waters
HSST3 C18 1.8 [tm, 50 mm x 2.1 mm; eluent A: water + 0.01% formic acid; eluent
B: acetonitrile
+ 0.01% formic acid; gradient: 0.0 min 2% B -> 0.5 min 2% B -> 7.5 min 95% B -
> 10.0 min 95%
B; oven: 50 C; flow rate: 1.00 ml/min; UV detection: 210-400 nm.
Method 17: Instrument: Waters TOF instrument; UPLC: Waters Acquity I-CLASS;
column: Waters
Acquity UPLC Peptide BEH C18, 300 A, 1.7 [tm; 150 mm x 2.1 mm; eluent A: water
+ 0.01% formic
acid, eluent B: acetonitrile + 0.01% formic acid; gradient: 0.0 min 90% A ->
0.25 min 90% A ->
8.0 min 45% A -> 10.0 min 2% A -> 12.0 min 2% A; oven: 50 C; flow rate: 0.475
ml/min; UV
detection: 210 nm.
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Method 18: Instrument: Waters TOF instrument; UPLC: Waters Acquity I-CLASS;
column: Waters
Acquity UPLC HSS T3, 1.8 [tm, 50 mm x 1 mm; eluent A: water + 0.01% formic
acid, eluent B:
acetonitrile + 0.01% formic acid; gradient: 0.0 min 95% A ¨> 6.0 min 5% A ¨>
7.5 min 5% A; oven:
50 C; flow rate: 0.35 ml/min; UV detection: 210 nm.
Method 19: Instrument: Shimadzu LCMS-2020; column: Poroshell HPH C18 2.7 [tm,
50 mm x
3.0 mm; eluent A: water 6.5 mM ammonium carbonate, eluent B: acetonitrile,
gradient: 0.0 min 10%
B ¨*2.1 min 95% B ¨> 2.7 min 95% B ¨> 2.75 min 10% B; oven: 40 C; flow rate:
1.2 ml/min; UV
detection: 190-400 nm.
Microwave: The microwave reactor used was a "single-mode" instrument of the
EmrysTm Optimizer
type.
When compounds according to the invention are purified by preparative HPLC by
the above -
described methods in which the eluents contain additives, for example
trifluoroacetic acid, formic
acid or ammonia, the compounds according to the invention may be obtained in
salt form, for
example as trifluoroacetate, formate or ammonium salt, if the compounds
according to the invention
contain a sufficiently basic or acidic functionality. Such a salt can be
converted to the corresponding
free base or acid by various methods known to the person skilled in the art.
In the case of the synthesis intermediates and working examples of the
invention described
hereinafter, any compound specified in the form of a salt of the corresponding
base or acid is
generally a salt of unknown exact stoichiometric composition, as obtained by
the respective
preparation and/or purification process. Unless specified in more detail,
additions to names and
structural formulae, such as "hydrochloride", "trifluoroacetate", "sodium
salt" or "x HC1", "x
CF3COOH", "x Nat" should not therefore be understood in a stoichiometric sense
in the case of such
salts, but have merely descriptive character with regard to the salt-forming
components present
therein.
This applies correspondingly if synthesis intermediates or working examples or
salts thereof were
obtained in the form of solvates, for example hydrates, of unknown
stoichiometric composition (if
they are of a defined type) by the preparation and/or purification processes
described.
In NMR spectra of mixtures of stereoisomers, numbers mentioned with "/"
indicate that the
stereoisomers show separate signals for the respective hydrogen atom, i.e.
"..../ (2s, 1H)" means
that one hydrogen atom is represented by 2 singlets, each singlet from one or
more different
stereoisomer(s).
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General Synthesis Methods
General Method 1: Etherification using sodium hydride
The respective alcohol (1.0 eq.) and the respective bromide (1.0-1.25 eq.)
were dissolved in
tetrahydrofuran (0.2-0.4 M) and cooled to 0 C. Then sodium hydride (60%
dispersion in mineral oil,
1.5-3.0 eq.) was added and stirring was continued for 1-3 hat 0 C and
overnight at RT. The reaction
mixture was diluted with ethyl acetate and saturated aqueous ammonium chloride
solution,
subsequently washed with water and saturated aqueous sodium chloride solution,
dried over
anhydrous sodium sulfate and filtered. The filtrate was concentrated under
reduced pressure and the
crude product was either purified by column chromatography (silica gel,
eluent: cyclohexane / ethyl
acetate mixtures or dichloromethane! methanol mixtures) or preparative HPLC
(reversed phase,
eluent: water! acetonitrile gradients or water! methanol gradients).
General Method 2: Mitsunobu reaction using
(tributylphosphoranylidene)acetonitrile
(Tributylphosphoranylidene)acetonitrile (1.5-3.0 eq.) was added to a solution
of the respective
primary or secondary alcohol (1.0 eq.) and the respective phenol (1.0-1.5 eq.)
in toluene (0.1-0.5 M)
in a microwave vessel. The reaction vessel was then sealed, placed into a
microwave reactor,
irradiated at 100 to 160 C and stirred at this temperature for 1-8 h. The
reaction mixture was then
cooled to RT and concentrated under reduced pressure. The crude product was
either purified by
column chromatography (silica gel, eluent: cyclohexane / ethyl acetate
mixtures or dichloromethane
/ methanol mixtures) or preparative HPLC (reversed phase, eluent: water!
acetonitrile gradients or
water / methanol gradients).
General Method 3a: Ring closure using Miyaura borylation reaction and Suzuki
reaction
The respective ether (1.0 eq.), bis(pinacolato)diboron (1.25-2.0 eq.),
palladium(II) acetate (0.03-0.1
eq.), tricyclohexylphosphine (0.1-0.15 eq.) and potassium acetate (3.0-4.5
eq.) were dissolved in
N,N-dimethylformamide (0.25-0.4 M) and argon was passed through the resulting
suspension for 10
min. The reaction mixture was heated to 80-100 C and stirred at this
temperature overnight, followed
by the addition of saturated aqueous sodium bicarbonate solution (5-7 ml/mmol
of the respective
ether) and tetrakis(triphenylphosphine)palladium(0) (0.03-0.05 eq.). Stirring
was then continued at
80-100 C for additional 2-72 h and the reaction mixture was either filtered
over silica gel and eluted
with dichloromethane or extracted with ethyl acetate, dried over anhydrous
sodium sulfate and
filtered. The filtrate was concentrated under reduced pressure and the crude
product was either
purified by column chromatography (silica gel, eluent: cyclohexane / ethyl
acetate mixtures or
dichloromethane / methanol mixtures) or preparative HPLC (reversed phase,
eluent: water /
acetonitrile gradients or water / methanol gradients).
General Method 3b: Ring closure using CH-activation
The respective ether (1.0 eq.) and potassium pivalate or potassium acetate
(2.0-6.0 eq.) were
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dissolved in N,N-dimethylacetamide (0.01-0.1 M) and argon was passed through
the mixture for 10
min. Subsequently, tetrakis(triphenylphosphine)palladium(0) (0.05-0.2 eq.) was
added and the
mixture was stirred for 2-48 h at 100-150 C. The residue was diluted with
water and extracted with
diethyl ether. The combined organic phases were washed with brine, dried over
anhydrous
magnesium sulfate, filtered and concentrated under reduced pressure. The crude
product was either
purified by column chromatography (silica gel, eluent: cyclohexane / ethyl
acetate mixtures or
dichloromethane / methanol mixtures) or preparative HPLC (reversed phase,
eluent: water /
acetonitrile gradients or water / methanol gradients).
General Method 4: Pyridinone formation using acetic acid and sodium iodine
The respective 2-methoxypyridine derivative (1.0 eq.) was dissolved in acetic
acid (0.05-0.2 M),
sodium iodide (2.0-4.0 eq.) was added and the resulting mixture was stirred at
80-100 C for 2-24 h.
Then dichloromethane and saturated aqueous sodium bicarbonate solution were
added and the
aqueous phase was extracted with dichloromethane. The combined organic phases
were washed with
saturated aqueous sodium chloride solution, dried over anhydrous sodium
sulfate, filtered and
concentrated under reduced pressure. The crude product was then purified
either by column
chromatography (silica gel, eluent: cyclohexane / ethyl acetate mixtures or
dichloromethane /
methanol mixtures) or preparative HPLC (reversed phase, eluent: water /
acetonitrile gradients or
water / methanol gradients) or used in the subsequent reaction without further
purification.
General Method 5: Pyridinone formation using 4-toluenesulfonic acid
monohydrate and
.. lithium iodine
The respective 2-methoxypyridine derivative (1.0 eq.) was dissolved in 1-
butanol (0.1-0.25 M), 4-
toluenesulfonic acid monohydrate (1.5-2.5 eq.) and lithium iodide (5.0-10.0
eq.) were added and the
resulting mixture was stirred at 80-100 C for 2-6 h. The reaction mixture was
then concentrated
under reduced pressure and the residue was partitioned between ethyl acetate
and water. The organic
layer was separated, washed with saturated aqueous sodium chloride solution,
dried over anhydrous
sodium sulfate, filtered and concentrated under reduced pressure. The crude
product was purified
either by column chromatography (silica gel, eluent: cyclohexane / ethyl
acetate mixtures or
dichloromethane / methanol mixtures) or preparative HPLC (reversed phase,
eluent: water /
acetonitrile gradients or water / methanol gradients) or used in the
subsequent reaction without
.. further purification.
General Method 6: Pyridinone formation using ethanethiol and sodium hydride
The respective 2-methoxypyridine derivative (1.0 eq.) was dissolved in N,N-
dimethylformamide
(0.1-0.25 M) followed by the addition of ethanethiol (6.0-10.0 eq.). The
mixture was cooled to 0 C,
sodium hydride (60% dispersion in mineral oil, 3.0-5.0 eq.) was added,
stirring was continued at 0 C
for 10-20 min and then heated to 80-100 C overnight. Subsequently, water was
added, the mixture
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was neutralized to pH 7 by slow addition of aqueous hydrochloric acid (1 N)
and extracted with ethyl
acetate. The combined organic phases were dried over anhydrous magnesium
sulfate, filtered and
concentrated under reduced pressure. The crude product was purified by
preparative HPLC (reversed
phase, eluent: water / acetonitrile gradients or water / methanol gradients).
General Method 7: Alkylation of pyridinone derivatives with a-bromo-ester and
a-bromo-
amide derivatives
The respective pyridinone derivative (1.0 eq.) was dissolved in a 4:1 mixture
of 2-propanol and
acetone (0.05-0.15 M) and 1,1,3,3-tetramethylguanidine (3.0-5.0 eq.) was added
at RT. After stirring
or shaking for 15 min, the respective a-bromo-ester or a-bromo-amide
derivative (1.0-2.5 eq.) was
added and stirring or shaking was continued overnight at RT. The crude mixture
was then directly
purified by preparative HPLC or concentrated under reduced pressure and the
crude mixture was
purified by preparative HPLC (reversed phase, eluent: water / acetonitrile
gradients or water /
methanol gradients).
General Method 8: Synthesis of triflate derivatives
A solution of trifluoromethanesulfonic anhydride (1.1-1.3 eq.) in
dichloromethane (0.1-0.25 M)
under argon atmosphere was cooled to -78 C and a solution of the corresponding
alcohol (1.0 eq.)
and trimethylamine or pyridine (1.1-1.5 eq.) in dichloromethane (0.75-1.0 M)
was added slowly.
Stirring was continued at -78 C for 0.5-2 h before the mixture was warmed to
RT, diluted with methyl
tert-butyl ether and washed with a 3:1 mixture of a saturated aqueous sodium
chloride solution and
aqueous hydrochloric acid (1 N). The organic phase was then dried over
anhydrous sodium sulfate,
filtered and concentrated under reduced pressure to obtain the crude product,
which was used in the
subsequent reaction without further purification.
General Method 9: a-Alkylation of N-substituted pyridinone derivatives with
triflate or
bromide derivatives
Under argon atmosphere, the respective N-substituted pyridinone derivative
(1.0 eq.) was dissolved
in tetrahydrofuran (0.1-0.2 M). At -78 C, a solution of sodium
bis(trimethylsilyl)amide in
tetrahydrofuran (1 M, 1.15-1.5 eq.) was added dropwise. After 20-30 min, the
respective triflate or
bromide derivative (1.15-1.5 eq.) dissolved in tetrahydrofuran (0.2-0.4 M) was
added slowly. The
mixture was stirred at -78 C for 15-30 min and for 0.5-1.0 h at RT. Then the
mixture was cooled to
-78 C before glacial acetic acid (1.5-3.0 eq.) was added. The reaction mixture
was concentrated
under reduced pressure and the crude product was either purified by column
chromatography (silica
gel, eluent: cyclohexane / ethyl acetate mixtures or dichloromethane /
methanol mixtures) or
preparative HPLC (reversed phase, eluent: water / acetonitrile gradients or
water / methanol
gradients) or used in the subsequent reaction without further purification.
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General Method 10: Ester cleavage using lithium hydroxide
The respective ester (1.0 eq.) was dissolved in tetrahydrofuran (0.05-0.15 M)
and an aqueous solution
of lithium hydroxide or sodium hydroxide (0.3-1.0 M, 5.0-10.0 eq.) was added.
The reaction mixture
was stirred at RT for 2 h up to overnight and acidified by addition of aqueous
solution of hydrochloric
acid (1 N). The resulting crude product was filtered and subsequently washed
with water and
cyclohexane or directly concentrated under reduced pressure. The residue was
then purified by
preparative HPLC (reversed phase, eluent: water / acetonitrile gradients or
water / methanol
gradients) or used in the subsequent reaction without further purification.
General Method 11: Amide coupling using T3P/pyridine
To a solution of the respective carboxylic acid (1 eq.) and the respective
amine (1.1-1.5 eq.) in
pyridine (about 0.1 M) was added T3P (50% solution in N,N-dimethylformamide or
in ethyl acetate,
1.5-4.0 eq.) at RT and the mixture was then stirred at RT or heated to 50-80
C. Alternatively, to a
solution of the respective carboxylic acid (1.0 eq.) in pyridine (about 0.1 M)
was added T3P (50%
solution in N,N-dimethylformamide or in ethyl acetate, 1.5-4 eq.) and the
solution was stirred for 1
to 10 min at RT. The respective amine (1.1-1.5 eq.) was then added and the
reaction mixture was
stirred at RT or heated to 50-80 C. After stirring at the respective
temperature for 1-48 h, the reaction
mixture was cooled to RT and either directly concentrated under reduced
pressure or diluted with
water, extracted with ethyl acetate, dried over anhydrous sodium sulfate,
filtered and concentrated
under reduced pressure. The crude product was purified either by column
chromatography
(cyclohexane / ethyl acetate mixtures or dichloromethane / methanol mixtures)
or preparative HPLC
(reversed phase, eluent: water / acetonitrile gradients or water / methanol
gradients).
Starting compounds
Example 1.1A
4-{ R2R)-2 -Bromobutanoyl] amino -2 -fluorobenzamide (single stere oi some r)
H 3
.rN Br F
0 0
N H 2
(2R)-2-Bromobutanoic acid (single stereoisomer) (3.5 g, 21.2 mmol, 1.1 eq.),
pyridine (1.7 ml,
21.2 mmol, 1.1 eq.) and T3P (17.2 ml, 50% solution in ethyl acetate, 28.9
mmol, 1.5 eq.) were added
under argon atmosphere at 0-5 C to a suspension of 4-amino-2-fluorobenzamide
(3.0 g, 19.3 mmol)
in tetrahydrofuran (30 m1). The reaction mixture was allowed to warm to RT and
stirred for 30 min.
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The reaction mixture was then cooled to 10 C, mixed dropwise with water (35
ml), stirred for 15
min, followed by the addition of further water (25 ml), and stirred for 30
min. The forming precipitate
was filtered, washed with water and dried in vacuo. Yield: 5.23 g (90% of
theory). From the
combined filtrates, further precipitate formed which was filtered, washed with
water and dried in
vacuo. Yield: 0.5 g (9% of theory).
both batches: LC-MS (method 4): Rt = 1.25 min; MS (ESIpos): m/z = 303 [M+I-11+
Example 1.2A
4- [(2R)-2 -Bromobutanoyl] amino I benzamide (single stereoisomer)
H 3
H
Br" yN
0 0
N H 2
(2R)-2-Bromobutanoic acid (single stereoisomer) (3.97 g, 23.8 mmol, 1.1 eq.)
was added slowly
under argon atmosphere to an ice-cooled suspension of 4-aminobenzamide (3.00
g, 21.6 mmol) in
tetrahydrofuran (22 ml), followed by the addition of pyridine (1.9 ml, 23.8
mmol, 1.1 eq.) and T3P
(19 ml, 50% solution in ethyl acetate, 32.4 mmol, 1.5 eq.) at a temperature of
0-5 C. The reaction
mixture was allowed to warm to RT and stirred at RT for 30 min. Water (35 ml)
was added dropwise
at 10 C. The reaction mixture was stirred for 15 min, followed by addition of
further water (25 ml),
and stirred for another 30 min. The forming precipitate was filtered, washed
with water and dried in
vacuo. The precipitate was suspended in a mixture of dichloromethane / ethyl
acetate, filtered (the
filtrate was discarded), suspended in tetrahydrofuran, filtered and dried in
vacuo. Yield: 5.72 g (93%
of theory). Further crystallization from the second filtrate yielded another
210 mg (3% of theory).
main batch: LC-MS (method 4): R1= 1.12 min; MS (ESIpos): m/z = 285 [M+I-11+
main batch: 'H-NMR (400 MHz, DM50-c/6): 6 [ppm] = 10.52 (s, 1H), 7.94-7.81 (m,
3H), 7.66 (d,
2H), 7.26 (br s, 1H), 4.48 (t, 1H), 2.17-1.88 (m, 2H), 0.96 (t, 3H).
Example 1.3A
(2R)-2-Bromo-N-(2-methyl-2H-benzotriazol-5-y1)butanamide (single stereoisomer)
H 3
H
N
Br -r
N¨C H 3
0
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(2R)-2-Bromobutanoic acid (single stereoisomer) (695 mg, 4.16 mmol, 1.1 eq.)
was added slowly
under argon atmosphere to an ice-cooled suspension of 2-methyl-2H-benzotriazol-
5-amine (590 mg,
3.78 mmol) in tetrahydrofuran (3.8 ml), followed by the addition of pyridine
(337 IA, 4.16 mmol,
1.1 eq.) and T3P (3.38 ml, 50% solution in ethyl acetate, 5.67 mmol, 1.5 eq.)
at a temperature of 0-
5 C. The reaction mixture was allowed to warm to RT and stirred at RT for 30
min. Water (25 ml)
was added dropwise at 10 C. The reaction mixture was stirred for 15 min,
followed by addition of
ethyl acetate (15 m1). After phase separation, the aqueous phase was extracted
with ethyl acetate.
The combined organic phases were washed with brine (15 ml), dried over
anhydrous magnesium
sulfate, filtered and concentrated under reduced pressure. Yield: 980 mg (85%
of theory).
.. LC-MS (method 1): Rt = 0.82 min; MS (ESIpos): m/z = 297 [M+H1+
'H-NMR (400 MHz, DM50-d6): 6 [ppm] = 10.55 (s, 1H), 8.36 (d, 1H), 7.89 (d,
1H), 7.43 (dd, 1H),
4.51 (t, 1H), 4.46 (s, 3H), 2.19-1.91 (m, 2H), 0.98 (t, 3H).
Example 1.4A
(2R)-2-Bromo-N{2-(difluoromethyl)-2H-indazol-5-yllbutanamide (single
stereoisomer)
/C H 3
H
N
Br
0
F H
General Method 11 was carried out with 2-(difluoromethyl)-2H-indazol-5-amine
hydrochloride
(24.0 mg, 109 umol, 1.0 eq.), (2R)-2-bromobutanoic acid (single stereoisomer)
(20.1 mg, 120 umol,
1.1 eq.), pyridine (9.7 IA, 120 umol, 1.1 eq.) and T3P (98 IA, 50% solution in
ethyl acetate, 160 umol,
1.5 eq.) in tetrahydrofuran (300 ul) including the following variations of the
procedure: The mixture
was stirred at 0 C for the addition of the reagents and at RT for 30 min.
Water was added at 10 C to
the reaction mixture and stirring was continued for 15 min. The crude mixture
was extracted with
dichloromethane, dried over anhydrous sodium sulfate, filtered, concentrated
under reduced pressure
and used in the subsequent reaction without further purification. Yield: 59.0
mg (quantitative of
theory).
LC-MS (method 5): R1= 1.12 min; MS (ESIpos): m/z = 332 [M+H1+
Example 1.5A
(2R)-2-Bromo-N{2-(trifluoromethyl)-2H-indazol-5-yllbutanamide (single
stereoisomer)
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H 3
E H
Br N
General Method 11 was carried out with 2-(trifluoromethyl)-2H-indazol-5-amine
hydrochloride
(65.3 mg, 272 lama 1.0 eq.), (2R)-2-bromobutanoic acid (single stereoisomer)
(50.0 mg, 299 lama
1.1 eq.), pyridine (24 [11, 300 lama 1.1 eq.) and T3P (240 [11, 50% solution
in ethyl acetate, 410
lama 1.5 eq.) in tetrahydrofuran (500 [I1) including the following variations
of the procedure: The
mixture was stirred at 0 C for the addition of the reagents and at RT for 30
min. Subsequently,
additional amounts of (2R)-2-bromobutanoic acid (22.7 mg, 136 lama 0.5 eq.)
and T3P (81 [11, 50%
solution in ethyl acetate, 136 lama 0.5 eq.) were added and stirring was
continued for 30 min. Water
was added at 10 C to the reaction mixture and stirring was continued for 15
min. The crude mixture
was extracted with dichloromethane, dried over anhydrous sodium sulfate,
filtered, concentrated
under reduced pressure and used in the subsequent reaction without further
purification. Yield: 181
mg (quantitative of theory).
LC-MS (method 5): R1= 1.25 min; MS (ESIpos): m/z = 350 [M+F11+
Example 1.6A
Ethyl 3-cyclobutylpropanoate
c(30 H 3
0
To a solution of 3-cyclobutylpropanoic acid (1.00 g, 7.80 mmol, 1.0 eq.) in
ethanol (15 ml) was
added thionyl chloride (850 IA, 12 mmol, 1.5 eq.) at 0 C. The resulting
mixture was then warmed to
RT and stirred overnight. The crude mixture was diluted with ethyl acetate,
washed with saturated
aqueous solution of sodium bicarbonate and extracted with ethyl acetate. The
combined organic
phases were dried over anhydrous sodium sulfate, filtered and concentrated
under reduced pressure.
The crude product was used in the subsequent reaction without further
purification. Yield: 1.02 g
(84% of theory).
GC-MS (method 6): R1= 3.15 min; MS (APCIpos): m/z = 157 [M+I-11+
Example 1.6B
Ethyl 2-bromo-3-cyclobutylpropanoate (racemate)
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c1:::70 C H
Br 3
0
To a solution of diisopropylamine (1.3 ml, 9.0 mmol, 1.4 eq.) in
tetrahydrofuran (20 ml) was added
a solution of n-butyllithium (3.1 ml, 2.5 M in hexane, 7.7 mmol, 1.2 eq.) at 0
C. After stirring at this
temperature for 30 min, the mixture was cooled to -78 C and a solution of
ethyl 3-
cyclobutylpropanoate (1.00 g, 6.40 mmol, 1.0 eq.) in tetrahydrofuran (20 ml)
was added dropwise.
The mixture was stirred at -78 C for 45 min, followed by the addition of 1,2-
dibromo-1,1,2,2-
tetrachloroethane (2.50 g, 7.68 mmol, 1.2 eq.) in tetrahydrofuran (10 m1). The
resulting mixture was
allowed to warm to RT over a period of 1 h, quenched by the addition of
saturated aqueous solution
of ammonium chloride and extracted with dichloromethane. The combined organic
layers were dried
over anhydrous sodium sulfate, filtered and concentrated under reduced
pressure. The residue was
purified by column chromatography (silica gel, eluent: cyclohexane /ethyl
acetate 98:2 to 9:1). Yield:
540 mg (41% purity, 15% of theory).
GC-MS (method 6): Rt = 4.33 min; MS (APCIpos): m/z = 235 [M+I-11+
Example 1.6C
2-Bromo-3-cyclobutylpropanoic acid (racemate)
c Br H
0
General Method 10 was carried out with ethyl 2-bromo-3-cyclobutylpropanoate
(racemate) (530 mg,
41% purity, 924 [Imo', 1.0 eq.) and lithium hydroxide (111 mg, 4.62 mmol, 5.0
eq.) in a mixture of
tetrahydrofuran / water (2:1, 9 ml) including the following variations of the
procedure: The reaction
mixture was stirred at RT for 2 h, then acidified by the addition of aqueous
hydrochloric acid (1 N)
and extracted with ethyl acetate. The combined organic phases were dried over
anhydrous sodium
sulfate, filtered and concentrated under reduced pressure. The crude product
was used in the
subsequent reaction without further purification.
Example 1.6D
4-(2-Bromo-3-cyclobutylpropanamido)-2-fluorobenzamide (racemate)
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Br 1 1 1 I 7IN
0 N H 2
0
General Method 11 was carried out two times with crude product of 2-bromo-3-
cyclobutylpropanoic
acid (racemate) (50.0 mg, 0.24 mmol, 1.0 eq.), 4-amino-2-fluorobenzamide (55.8
mg, 0.36 mmol,
1.5 eq.) and T3P (0.56 ml, 50% solution in ethyl acetate, 0.97 mmol, 4.0 eq.)
in pyridine (3 ml) and
crude product of 2-bromo-3-cyclobutylpropanoic acid (racemate) (410 mg, 1.98
mmol, 1.0 eq.),
4-amino-2-fluorobenzamide (458 mg, 2.97 mmol, 1.5 eq.) and T3P (4.6 ml, 50%
solution in ethyl
acetate, 7.9 mmol, 4.0 eq.) in pyridine (20 ml) including the following
variations of the procedure:
The two reaction mixtures were combined, washed with saturated aqueous
solution of ammonium
chloride and water, dried over anhydrous sodium sulfate, filtered and
concentrated under reduced
pressure. The crude mixture was purified by preparative HPLC (reversed phase,
eluent: water with
0.05% formic acid! acetonitrile 90:10 to 5:95). Yield: 33.0 mg (91% purity, 4%
of theory).
LC-MS (method 5): R1= 1.16 min; MS (ESIpos): m/z = 343 [M+F11+
Example 1.7A
tert-Butyl 3-(tetrahydro-2H-pyran-2-yl)propanoate (racemate)
H 3
hC H3
0 C H 3
Tetrahydro-2H-pyran-2-ylmethanol (racemate) (20.0 g, 172 mmol, 1.0 eq.), tert-
butyl acetate
(230 ml, 1.7 mol, 10.0 eq.), bis(1,5-cyclooctadiene)diiridium(I) dichloride
(5.78 g, 8.61 mmol,
0.1 eq.), triphenylphosphine (6.77 g, 25.8 mmol, 0.2 eq.) and potassium tert-
butanolate (38.6 g,
344 mmol, 2.0 eq.) were dissolved in degassed tert-butanol (200 ml) and
stirred at 100 C for 6 days.
The reaction mixture was filtered and washed repeatedly with ethyl acetate.
After removal of the
volatiles under reduced pressure, the crude mixture was purified by column
chromatography (silica
gel, eluent: cyclohexane / ethyl acetate with 0.1% N,N-diisopropylethylamine:
100:0 to 98:2 to 96:4).
Yield: 8.40 g (23% of theory).
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1H-NMR (400 MHz, DMSO-d6): 6 [ppm] = 3.88-3.80 (m, 1H), 3.32-3.24 (m, 1H),
3.20-3.13 (m,
1H), 2.29-2.13 (m, 2H), 1.78-1.67 (m, 1H), 1.63-1.49 (m, 3H), 1.47-1.35 (m,
3H), 1.39 (s, 9H), 1.18-
1.06 (m, 1H).
Example 1.7B
tert-Butyl 2-bromo-3-(tetrahydro-2H-pyran-2-yl)propanoate (mixture of
stereoisomers)
B r hC
C H 3
H3
0 CH3
N,N-Diisopropylamine (7.7 ml, 55 mmol, 1.4 eq.) was dissolved in
tetrahydrofuran (120 ml) and
cooled to 0 C before a solution of n-butyllithium (19 ml, 2.5 M in hexane, 47
mmol, 1.2 eq.) was
added. The mixture was stirred at 0 C for 30 min, then cooled to -78 C,
followed by the dropwise
addition of tert-butyl 3-(tetrahydro-2H-pyran-2-yl)propanoate (racemate) (8.40
g, 39.2 mmol, 1.0
eq.) in tetrahydrofuran (62 m1). Stirring was continued at -78 C for 45 min,
1,2-dibromo-1,1,2,2-
tetrafluoroethane (12.2 g, 47.0 mmol, 1.2 eq.) in tetrahydrofuran (62 ml) was
added dropwise and
the mixture was warmed to RT over 1 h. After addition of saturated aqueous
solution of ammonium
chloride, the aqueous phase was extracted with dichloromethane. The combined
organic phases were
dried over sodium sulfate, filtered and concentrated under reduced pressure.
The crude product was
purified by column chromatography (silica gel, eluent: cyclohexane / ethyl
acetate with 0.1% N,N-
diisopropylethylamine: 100:0 to 96:4 to 70:30). Yield: 7.00 g (61% of theory).
GC-MS (method 6): Rt = 5.38 / 5.50 min; MS (APCIpos): m/z = 239 [M-tBu+H1+
Example 1.7C
2-Bromo-3-(tetrahydro-2H-pyran-2-yl)propanoic acid (mixture of stereoisomers)
0
0H
Br
0
tert-Butyl 2-bromo-3-(tetrahydro-2H-pyran-2-yl)propanoate (mixture of
stereoisomers) (7.00 g,
23.9 mmol, 1.0 eq.) was dissolved in trifluoroacetic acid (140 ml) and stirred
at RT for 2 h. The
reaction mixture was concentrated under reduced pressure and coevaporated
three times with toluene.
The crude product was used in the subsequent reaction without further
purification. Yield: 5.65 g
(quantitative of theory).
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GC-MS (method 6): Rt = 5.53 min; MS (APCIpos): m/z = 239 [M+1-11+
Example 1.7D
4-( (2R)-2-Bromo-34(25)-tetrahydro-2H-pyran-2-yllpropanoyl amino)-2-
fluorobenzamide (single
stereoisomer)
H
N
Br(
0 NH 2
General Method 11 was carried out with 4-amino-2-fluorobenzamide (3.68 g, 23.9
mmol, 1.0 eq.),
2-bromo-3-(tetrahydro-2H-pyran-2-yl)propanoic acid (mixture of stereoisomers)
(5.66 g, 23.9 mmol,
1.0 eq.), pyridine (2.1 ml, 26 mmol, 1.1 eq.) and T3P (21 ml, 50% solution in
ethyl acetate, 36 mmol,
1.5 eq.) in tetrahydrofuran (43 ml) including the following variations of the
procedure: After stirring
at RT overnight, the reaction mixture was worked up by addition of water,
extracted with ethyl
acetate, washed with brine, dried and concentrated under reduced pressure.
Stereoisomer separation
of the crude mixture gave:
single stereoisomer 1 (chiral SFC: R1= 1.52 min): 1.87 g,
single stereoisomer 2 (the title compound 1.7D) (chiral SFC: Rt = 1.66 min,
99% de): 1.90 g (21%
of theory),
single stereoisomer 3 (chiral SFC: R1= 1.30 min): 1.18 g,
single stereoisomer 4 (chiral SFC: R1= 1.34 min): 1.15 g.
Separation method 1: SFC: column: Daicel Chiralpak AD 20 um, 450 mm x 50 mm;
eluent: 75%
carbon dioxide /25% 2-propanol; temperature: 40 C; flow rate: 400 ml/min; UV
detection: 210 nm.
Separation method 2: Single stereoisomer 3 and single stereoisomer 4 eluted as
a mixture in the first
separation. This mixture was then separated according to the following
conditions: SFC: column:
Daicel Chiralpak AD 20 um, 450 mm x 50 mm; eluent: 20% carbon dioxide / 80% 2-
propanol;
temperature: 40 C; flow rate: 400 ml/min; UV detection: 210 nm.
Analysis method: SFC: column: Daicel Chiralpak AD-H 5 um, 250 mm x 4.6 mm;
eluent: 80%
carbon dioxide / 20% methanol; flow rate: 3.0 ml/min; UV detection: 210 nm.
LC-MS (method 1): Rt = 0.82 min; MS (ESIpos): m/z = 373 [M+1-11+
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1H-NMR (500 MHz, DMSO-d6): 6 [ppm] = 10.71 (s, 1H), 7.72-7.66 (m, 1H), 7.65-
7.59 (m, 1H),
7.57-7.46 (m, 2H), 7.43-7.26 (m, 1H), 4.80-4.66 (m, 1H), 3.88 (d, 1H), 3.54-
3.41 (m, 1H), 3.40-3.33
(m, 1H), 2.11-1.95 (m, 2H), 1.88-1.69 (m, 1H), 1.61 (d, 1H), 1.53-1.38 (m,
3H), 1.31-1.21 (m, 1H).
Example 1.8A
4-( (2R)-2-Bromo-3-[(25)-tetrahydro-2H-pyran-2-yllpropanoyl}amino)benzamide
(single
stereoisomer)
H
Br N
-r
0 NH 2
0
General Method 11 was carried out with 4-aminobenzamide (5.95 g, 43.7 mmol,
1.0 eq.), 2-bromo-
3-(tetrahydro-2H-pyran-2-yl)propanoic acid (mixture of stereoisomers) (11.4 g,
48.1 mmol, 1.1 eq.),
pyridine (3.9 ml, 48.1 mmol, 1.1 eq.) and T3P (39 ml, 50% solution in ethyl
acetate, 66 mmol, 1.5 eq.)
in tetrahydrofuran (79 ml) including the following variations of the
procedure: After stirring at RT
overnight, the reaction mixture was worked up by addition of water, extracted
with ethyl acetate,
washed with brine, dried and concentrated under reduced pressure. Stereoisomer
separation of the
crude mixture gave:
single stereoisomer 1 (the title compound 1.8A) (chiral SFC: Rt = 1.52 min,
99% de): 2.71 g (17%
of theory),
single stereoisomer 2 (chiral SFC: R1= 1.66 min): 3.29 g,
single stereoisomer 3 (chiral SFC: R1= 1.30 min): 1.43 g,
single stereoisomer 4 (chiral SFC: R1= 1.34 min): 1.45 g.
Separation method 1: SFC: column: Daicel Chiralpak AD 20 um, 450 mm x 50 mm;
eluent: 75%
carbon dioxide /25% 2-propanol; temperature: 40 C; flow rate: 400 ml/min; UV
detection: 210 nm.
Separation method 2: Single stereoisomer 3 and single stereoisomer 4 eluted as
a mixture in the first
separation. This mixture was then separated according to the following
conditions: SFC: column:
Daicel Chiralpak AD 20 um, 450 mm x 50 mm; eluent: 20% carbon dioxide / 80% 2-
propanol;
temperature: 40 C; flow rate: 400 ml/min; UV detection: 210 nm.
Analysis method: SFC: column: Daicel Chiralpak AD-H 5 um, 250 mm x 4.6 mm;
eluent: 80%
carbon dioxide / 20% methanol; flow rate: 3.0 ml/min; UV detection: 210 nm.
LC-MS (method 1): Rt = 0.75 min; MS (ESIpos): m/z = 355 [M+H1+
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1H-NMR (600 MHz, DMSO-d6): 6 [ppm] = 10.58 (s, 1H), 7.95-7.82 (m, 3H), 7.71-
7.63 (m, 2H),
7.29 (br s, 1H), 4.81 (dd, 1H), 3.92 (d, 1H), 3.58-3.43 (m, 1H), 3.42-3.37 (m,
1H), 2.12-1.99 (m, 2H),
1.80 (d, 1H), 1.64 (d, 1H), 1.57-1.41 (m, 4H), 1.37-1.22 (m, 1H).
Example 1.9A
tert-Butyl (4R)-4-hydroxypentanoate (single stereoisomer)
OH
cC H 3
0 H3
IC H3
o C H3
To a stirred solution of lithium diisopropylamide (86 ml, 2.0 M in
tetrahydrofuran / n-hexane,
170 mmol, 2.0 eq.) in tetrahydrofuran (250 ml) was added tert-butyl acetate
(20.0 g, 172 mmol,
2.0 eq.) dropwise at -78 C. After stirring for 0.5 h, the mixture was warmed
to -40 C,
diethylaluminum chloride (170 ml, 1.0 M in hexane, 170 mmol, 2.0 eq.) was
added over a period of
min and stirring was continued for further 15 min. Subsequently, (2R)-2-
methyloxirane (single
stereoisomer) (5.00 g, 86.1 mmol, 1.0 eq.) was added and the mixture was
stirred at -40 C for 5 h,
followed by the addition of saturated aqueous solution of ammonium chloride
and ice in aqueous
hydrochloric acid (6 N) at -20 C. The mixture was extracted with diethyl ether
and the combined
15 organic layers were washed with saturated aqueous solution of sodium
bicarbonate and brine, dried
over anhydrous sodium sulfate, filtered and concentrated under reduced
pressure. The residue was
purified by column chromatography (silica gel, eluent: n-hexane / ethyl
acetate 4:1). Yield: 8.0 g
(90% purity, 48% of theory).
1H-NMR (400 MHz, DM50-d6): 6 [ppm] = 4.43 (d, 1H), 3.62-3.49 (m, 1H), 2.33-
2.14 (m, 2H), 1.61-
20 1.46 (m, 2H), 1.39 (s, 9H), 1.03 (d, 3H).
Example 1.9B
tert-Butyl (4R)-4-methoxypentanoate (single stereoisomer)
C H 3
cCH3
0C H3
hC H3
0 CH3
To a suspension of tert-butyl (4R)-4-hydroxypentanoate (single stereoisomer)
(9.00 g, 95% purity,
49.1 mmol, 1.0 eq.) and freshly prepared silver(I) oxide (34.1 g, 147 mmol,
3.0 eq.) in
1,2-dichloroethane (150 ml) was added iodomethane (31 ml, 490 mmol, 10.0 eq.)
at 0 C. After
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stirring at 40 C for 72 h, the reaction mixture was filtered through a pad of
Celite and the filtrate
was concentrated under reduced pressure. The residue was purified by column
chromatography
(silica gel, eluent: n-hexane / ethyl acetate 8:1 to 1:1). Yield: 1.90 g (20%
of theory).
1H-NMR (400 MHz, DMSO-d6): 6 [ppm] = 3.28-3.21 (m, 1H), 3.18 (s, 3H), 2.20 (t,
2H), 1.71-1.50
(m, 2H), 1.39 (s, 9H), 1.04 (d, 3H).
Example 1.9C
tert-Butyl (4R)-2-bromo-4-methoxypentanoate (mixture of two diastereomers)
C H3
.../c Ci C H3
CH
Br
hCH3
0 CH3
To a solution of tert-butyl (4R)-4-methoxypentanoate (single stereoisomer)
(1.70 g, 8.58 mmol, 1.0
eq.) in tetrahydrofuran (70 ml) was added lithium diisopropylamide (6.4 ml,
2.0 M in
tetrahydrofuran, 13 mmol, 1.5 eq.) at -78 C. After stirring at -78 C for 15
min, a solution of 1,2-
dibromo-1,1,2,2-tetrachloroethane (6.98 g, 21.4 mmol, 2.5 eq.) in
tetrahydrofuran (10 ml) was added
dropwise. The resulting mixture was slowly allowed to warm to RT over a period
of 1 h and stirred
for further 2 h before water was added. The aqueous phase was extracted with
ethyl acetate and the
combined organic layers were washed with water and brine, dried over anhydrous
sodium sulfate,
filtered and concentrated under reduced pressure. The residue was purified by
preparative thin-layer
chromatography (silica gel, eluent: petroleum ether / ethyl acetate 8:1).
Yield: 0.95 g (90% purity,
37% of theory).
1H-NMR (400 MHz, DMSO-d6): 6 [ppm] = 4.40-4.28 (m, 1H), 3.47-3.36 / 3.34-3.26
(2m, 1H), 3.21 /
3.18 (2s, 3H), 2.21-1.86 (m, 2H), 1.43 / 1.42 (2s, 9H), 1.15-1.05 (m, 3H).
Example 1.10A
tert-Butyl (45)-4-hydroxypentanoate (single stereoisomer)
OH
H 3
0 H 3
H 3
C H 3
tert-Butyl acetate (4.00 g, 34.4 mmol, 2.0 eq.) was added dropwise at -78 C to
a stirred solution of
lithium diisopropylamide (17.2 ml, 34.4 mmol, 2 M in tetrahydrofuran / n-
hexane, 2.0 eq.) in
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tetrahydrofuran (50 m1). After stirring for 0.5 h, the mixture was warmed to -
40 C. Diethylaluminum
chloride (34.4 ml, 34.4 mmol, 1 Mm n-hexane, 2.0 eq.) was added over a period
of 5 min and stirring
was continued for further 15 min. Subsequently, (25)-2-methyloxirane (single
stereoisomer) (1.00 g,
17.2 mmol, 1.0 eq.) was added and the mixture was stirred at -40 C for 5 h,
followed by the addition
of saturated aqueous solution of ammonium chloride and ice in aqueous
hydrochloric acid (6 N) at -
20 C. The mixture was extracted with diethyl ether. The combined organic
layers were washed with
saturated aqueous solution of sodium bicarbonate and brine, dried over
anhydrous sodium sulfate,
filtered and concentrated under reduced pressure. The residue was purified by
column
chromatography (silica gel, eluent: n-hexane / ethyl acetate 4:1). Yield: 1.50
g (90% purity, 45% of
theory).
1H-NMR (400 MHz, DM50-d6): 6 [ppm] = 4.45 (d, 1H), 3.64-3.46 (m, 1H), 2.34-
2.12 (m, 2H), 1.61-
1.45 (m, 2H), 1.40 (s, 9H), 1.04 (d, 3H).
Example 1.10B
tert-Butyl (45)-4-methoxypentanoate (single stereoisomer)
C H 3
C'JC H 3
hC H 3
0 CH3
Iodomethane (61.3 g, 432 mmol, 10.0 eq.) was added at 0 C to a suspension of
tert-butyl (45)-4-
hydroxypentanoate (single stereoisomer) (8.00 g, 43.2 mmol, 1.0 eq.) and
freshly prepared silver(I)
oxide (30.0 g, 130 mmol, 3.0 eq.) in 1,2-dichloroethane (150 m1). After
stirring at 40 C for 72 h, the
reaction mixture was filtered through a pad of Celite and the filtrate was
concentrated under reduced
pressure. The residue was purified by column chromatography (silica gel,
eluent: n-hexane / ethyl
acetate 8:1 to 1:1). Yield: 2.20 g (26% of theory).
1H-NMR (400 MHz, DM50-d6): 6 [ppm] = 3.28-3.21 (m, 1H), 3.18 (s, 3H), 2.20 (t,
2H), 1.68-1.54
(m, 2H), 1.39 (s, 9H), 1.04 (d, 3H).
Alternative synthetic route:
NNN',AP-Tetramethylnaphthalene-1,8-diamine (14.36 g, 66.99 mmol, 1.1 eq.,
weighed out in glove
box!) was added under argon atmosphere at 0-4 C to a solution of tert-butyl
(4S)-4-
hydroxypentanoate (single stereoisomer) (13.10 g, 60.90 mmol) in
dichloromethane (260 m1). The
reaction mixture was stirred at RT for 30 min, cooled again to 0-4 C, mixed
with trimethyloxonium
tetrafluoroborate (19.91 g, 127.89 mmol, 2.1 eq., weighed out in glove box!),
stirred for 30 min at
0-4 C and then for 60 min while allowing to warm to RT. The reaction mixture
was quenched with
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water (250 ml) and diluted with dichloromethane (150 m1). The precipitate was
filtered off and
discarded. After phase separation, the aqueous phase was extracted two times
with dichloromethane.
The combined organic phases were washed with brine, dried over anhydrous
sodium sulfate, filtered
and concentrated under reduced pressure (at >100 mbar and <25 C). The residue
was purified by
column chromatography (silica gel, eluent: cyclohexane / ethyl acetate
gradient, eluent evaporation
at >80 mbar and <30 C). Yield: 10.13 g (88% of theory).
1H-NMR (600 MHz, DMSO-d6): 6 [ppm] = 3.28-3.22 (m, 1H), 3.19 (s, 3H), 2.20 (t,
2H), 1.64-1.57
(m, 2H), 1.39 (s, 9H), 1.05 (d, 3H).
Example 1.10C
tert-Butyl (45)-2-bromo-4-methoxypentanoate (mixture of two diastereomers)
0"
C Hq
H3
Br OC H3
hCH3
0 C H3
Lithium diisopropylamide (5.7 ml, 11.4 mmol, 2.0 M in tetrahydrofuran, 1.5
eq.) was added at -78 C
to a solution of tert-butyl (45)-4-methoxypentanoate (single stereoisomer)
(1.50 g, 7.6 mmol, 1.0
eq.) in tetrahydrofuran (60 m1). After stirring at -78 C for 15 min, a
solution of 1,2-dibromo-1,1,2,2-
tetrachloroethane (6.16 g, 18.9 mmol, 2.5 eq.) in tetrahydrofuran (8 ml) was
added dropwise. The
resulting mixture was slowly allowed to warm to RT over a period of 1 h and
stirred for further 2 h
before water was added. The aqueous phase was extracted with ethyl acetate.
The combined organic
layers were washed with water and brine, dried over anhydrous sodium sulfate,
filtered and
concentrated under reduced pressure. The residue was purified by preparative
thin-layer
chromatography (silica gel, eluent: petroleum ether / ethyl acetate 8:1).
Yield: 0.94 g (85% purity,
40% of theory).
1H-NMR (400 MHz, DMSO-d6): 6 [ppm] = 4.41-4.28 (m, 1H), 3.47-3.36 / 3.35-3.25
(2m, 1H), 3.21 /
3.18 (2s, 3H), 2.21-1.85 (m, 2H), 1.43 / 1.42 (2s, 9H), 1.15-1.05 (m, 3H).
Example 1.10D
.. (4S)-2-Bromo-4-methoxypentanoic acid (mixture of two diastereomers)
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CYCH3
Br OH
0
Trifluoroacetic acid (13.3 ml, 172.6 mmol, 20 eq.) was added dropwise under
argon atmosphere to
an ice-cooled solution of tert-butyl (45)-2-bromo-4-methoxypentanoate (mixture
of two
diastereomers) (2.7 g, 85% purity, 8.6 mmol) in dichloromethane (50 m1). The
reaction mixture was
.. stirred at RT for 2 h, followed by the addition of further trifluoroacetic
acid (3.3 ml, 43.2 mmol, 5.0
eq.). After stirring for another 1 h, the reaction mixture was concentrated in
vacuo and coevaporated
with dichloromethane. The residue was purified by column chromatography
(silica gel, eluent:
cyclohexane / ethyl acetate gradient). Yield: 1.37 g (75% of theory).
'H-NMR (600 MHz, DMSO-d6): 6 [ppm] = 4.41-4.36 (m, 1H), 3.48-3.40 / 3.35-3.28
(2m, 1H), 3.23 /
3.19 (2s, 3H), 2.17-2.02 / 1.96-1.89 (2m, 2H), 1.12 / 1.10 (2d, 3H).
Alternative synthetic route:
Lithium hydroxide monohydrate (233 g, 5.55 mol, 2.1 eq.) was added at RT to a
solution of methyl
(45)-2-bromo-4-methoxypentanoate (mixture of two diastereomers) (625 g, 94%
purity, 2.61 mol)
in a mixture of tetrahydrofuran / water (3:1, 6.11). The reaction mixture was
stirred at RT overnight
and mixed with 1 N aqueous hydrochloric acid (5 1). The aqueous phase was
extracted with 2-
methyltetrahydrofuran. The organic phase was dried and evaporated under
reduced pressure. The
crude material was used without further purification. Yield: 595 g (94%
purity, quantitative of
theory).
1H-NMR (600 MHz, DMSO-d6): 6 [ppm] = 12.9 (br s, 1H, two diastereomers), 4.44-
4.30 (m, 1H,
two diastereomers), 3.47-3.40 (m, 1H, major diastereomer), 3.37-3.27 (m, 1H,
minor diastereomer),
3.22 (s, 3H, major diastereomer), 3.18 (s, 3H, minor diastereomer), 2.17-2.00
(m, 2H, two
diastereomers), 1.11 (d, 3H, major diastereomer), 1.09 (d, 3H, minor
diastereomer).
Example 1.10E
4-{[(2R,45)-2-Bromo-4-methoxypentanoyllaminolbenzamide (single stereoisomer)
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CYCH3
)-CFi3
H
N
Brr
0 NH2
0
(45)-2-Bromo-4-methoxypentanoic acid (mixture of two diastereomers) (1.37 g,
6.5 mmol), pyridine
(579 [11, 7.2 mmol, 1.1 eq.) and T3P (5.7 ml, 50% solution in ethyl acetate,
9.8 mmol, 1.5 eq.) were
added under argon atmosphere at RT to a solution of 4-aminobenzamide (975 mg,
7.2 mmol, 1.1 eq.)
in tetrahydrofuran (25 m1). The reaction mixture was stirred at RT for 2 h,
quenched with water and
diluted with ethyl acetate. After phase separation, the aqueous phase was
extracted with ethyl acetate.
The combined organic phases were washed with brine, dried over anhydrous
sodium sulfate, filtered
and concentrated under reduced pressure. The residue was purified by column
chromatography
(silica gel, eluent: dichloromethane / methanol gradient). Yield: 1.91 g (86%
of theory).
LC-MS (method 4): R1= 1.18 min; MS (ESIpos): m/z = 329 [M+F11+
Diastereomer separation of 1910 mg of 4-{[(45)-2-bromo-4-
methoxypentanoyllamino}benzamide
(mixture of two diastereomers) gave
single stereoisomer 1 (the title compound Example 1.10E) (chiral HPLC: Rt =
11.7 min, >99% de):
901 mg,
single stereoisomer 2 (chiral HPLC: Rt = 14.0 min, 89% de): 576 mg.
Separation method: HPLC: column: Daicel Chiralpak IE 5 lam, 250 mm x 20 mm;
eluent: 80%
n-heptane / 20% ethanol; temperature: 40 C; flow rate: 15 ml/min; UV
detection: 220 nm.
Analysis method: HPLC: column: Daicel Chiralpak IE 5 lam, 250 mm x 4.6 mm;
eluent: 80%
iso-hexane / 20% ethanol; temperature: 40 C; flow rate: 1 ml/min; UV
detection: 220 nm.
'H-NMR (400 MHz, DM50-d6): 6 [ppm] = 10.56 (s, 1H), 7.92-7.81 (m, 3H), 7.65
(d, 2H), 7.25 (br
s, 1H), 4.75 (t, 1H), 3.54-3.44 (m, 1H), 3.26 (s, 3H), 2.10-2.02 (m, 2H), 1.15
(d, 3H).
Alternative synthetic route:
Under nitrogen atmosphere, 4-aminobenzamide (419 g, 3.08 mol, 1.17 eq.) in
tetrahydrofuran (6.11)
was treated at RT with (45)-2-bromo-4-methoxypentanoic acid (mixture of two
diastereomers)
(591 g, 94% purity, 2.63 mol), pyridine (249 ml, 3.08 mol, 1.17 eq.) and T3P
(50% in ethyl acetate,
2.51 1, 4.21 mol). After 4 h stirring at RT, the reaction mixture was diluted
with ethyl acetate (7.0 1)
and afterwards with water (8.0 1). After extraction and phase separation, the
aqueous phase was
reextracted with ethyl acetate (7.0 1). The combined organic phases were
washed with saturated
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aqueous sodium chloride solution (6.0 1). The residue was suspended in
diisopropyl ether (7.5 1) and
ethyl acetate (550 ml) and stirred for 2.5 h at RT. The mixture was filtered
under reduced pressure.
Methanol was added under stirring to allow for a good filtration. The obtained
solid was washed with
diisopropyl ether and dried under air atmosphere. This material was triturated
with acetonitrile at
80 C for 20 min. The suspension was slowly allowed to reach RT and then cooled
down to 0 C.
After stirring for 1 h, the suspension was filtered under reduced pressure,
the residue was washed
with cold acetonitrile and then dried under air atmosphere (yield: 81 g). The
filtrate from the first
filtration was concentrated under reduced pressure and the obtained residue
was triturated with
acetonitrile (5.0 1) at 80 C for 20 min. The suspension was slowly allowed to
reach RT and then
cooled down to 0 C. After stirring for 1 h, the suspension was filtered under
reduced pressure, the
residue was washed with cold acetonitrile and then dried under air atmosphere
(yield: 451 g). The
combined filtrates were concentrated under reduced pressure and the obtained
residue was dissolved
with acetonitrile (1.0 1) at 60 C. The suspension was slowly allowed to reach
RT and then cooled
down to 0 C. After stirring for 1 h, the suspension was filtered under reduced
pressure, the residue
was washed with cold acetonitrile and then dried under air atmosphere. This
material was dissolved
in dichloromethane and methanol and loaded onto Isolute . This material was
purified via column
chromatography (Biotage Isolera LS, 1.5 kg SNAP-Ultra cartridge, eluent:
dichloromethane / 20%
acetone (1 cv), 20% - 60% acetone gradient (10 cv), 60% acetone (2 cv) to
yield 63 g. Diastereomer
separation of all crude products (81 g, 451 g and 63 g) via SFC gave the
product as single
stereoisomer. Combined yield: 266 g (31% of theory, >99% de).
Separation method: SFC: column: Chiralpak AY, 20 um, 250 mm x 50 mm, eluent:
isocratic 82%
carbon dioxide / 18% methanol, 5 min, temperature: 30 C; flow rate: 350
ml/min; UV detection:
210 nm; backpressure 100 bar and SFC: column: Chiralpak AZ 20 um, 350 mm x 50
mm, eluent:
isocratic 65% carbon dioxide / 35% methanol; 5 min, temperature: 30 C; flow
rate: 350 ml/min; UV
detection: 210 nm; backpressure: 100 bar.
Chiral SFC: R1= 1.56 min, >99% de
Analysis method: SFC: column: Chiralcel AY-3, 3 um, 100 mm x 4.6 mm, eluent:
isocratic 80%
carbon dioxide / 20% methanol; 10 min, temperature: 40 C; flow rate: 3.0
ml/min; UV detection:
210 nm; backpressure: 130 bar.
LC-MS (method 4): R1= 1.16 min; MS (ESIpos): m/z = 329, 331 [M+I-11+
1H-NMR (600 MHz, DMSO-d6): 6 [ppm] = 10.5 (br s, 1H), 7.85 (br d, 3H), 7.65
(br d, 2H), 7.23 (br
s, 1H), 4.75 (br t, 1H), 3.57-3.43 (m, 1H), 3.26 (s, 3H), 2.07 (br t, 2H),
1.15 (br d, 3H).
Example 1.11A
tert-Butyl 4-oxobutanoate
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0
OC H3
hCH3
0 CH3
3,3,3-Triacetoxy-3-iodophthalide (123.7 g, 291.6 mmol, 2.0 eq.) was added in
portions at 0 C to a
mixture of tert-butyl 4-hydroxybutanoate (23.4 g, 145.8 mmol, 1.0 eq.) and
sodium bicarbonate
(24.5 g, 291.6 mmol, 2.0 eq.) in dichloromethane (500 m1). After stirring at
RT for 2 h, the reaction
mixture was quenched by the addition of a mixture of saturated aqueous
solution of sodium carbonate
and sodium thio sulfate (1:1), stirred for further 30 min and extracted with
dichloromethane. The
combined organic phases were washed with water, brine, dried over anhydrous
sodium sulfate,
filtered and concentrated under reduced pressure. The residue was purified by
column
chromatography (silica gel, eluent: petroleum ether / ethyl acetate 95:5).
Yield: 15.4 g (63% of
theory).
1H-NMR (300 MHz, CDC13): 6 [ppm] = 9.79 (s, 1H), 2.76-2.68 (m, 2H), 2.59-2.50
(m, 2H), 1.43 (s,
9H).
Example 1.11B
tert-Butyl 4-cyclopropy1-4-hydroxybutanoate (racemate)
0 H
c1C7 C H 3
hCH3
0 CH3
Cyclopropylmagnesium bromide (185.0 ml, 1.0 M in tetrahydrofuran, 185.0 mmol,
2.0 eq.) was
added under argon atmosphere at -10 C to a stirred solution of tert-butyl 4-
oxobutanoate (15.4 g,
92.5 mmol) in tetrahydrofuran (300 m1). The reaction mixture was allowed to
warm to RT within a
period of 1 h, stirred for another 1 h, quenched with aqueous solution of
aqueous hydrochloric acid
(2 N), diluted with ethyl acetate and washed with water and brine. The organic
phase was dried over
anhydrous sodium sulfate, filtered and concentrated under reduced pressure.
The residue was purified
by column chromatography (silica gel, eluent: petroleum ether / ethyl acetate
90:10). Yield: 5.88 g
(85% purity, 27% of theory).
1H-NMR (300 MHz, DMSO-d6): 6 [ppm] = 4.55-4.30 (m, 1H), 2.89-2.72 (m, 1H),
2.39-2.13 (m,
2H), 1.82-1.52 (m, 2H), 1.39 (s, 9H), 0.88-0.67 (m, 1H), 0.45-0.05 (m, 4H).
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Example 1.11C
tert-Butyl 4-cyclopropy1-4-methoxybutanoate (racemate)
CYCH3
cIC7LC H 3
hCH3
0 C H 3
Iodomethane (34.82 g, 245.3 mmol, 10.0 eq.) was added under argon atmosphere
at RT to a mixture
of tert-butyl 4-cyclopropy1-4-hydroxybutanoate (racemate) (5.78 g, 85% purity,
24.5 mmol) and
freshly prepared silver(I) oxide (17.05 g, 73.6 mmol, 3.0 eq.) in 1,2-
dichloroethane (100 m1). After
stirring at 45 C for 24 h, the resulting mixture was filtered through Celite .
The filtrate was
concentrated under reduced pressure. The residue was re-dissolved in 1,2-
dichloroethane (100 ml),
followed by the addition of silver(I) oxide (8.53 g, 36.8 mmol, 1.5 eq.) and
iodomethane (17.41 g,
.. 122.7 mmol, 5.0 eq.) under argon atmosphere at RT. The resulting mixture
was stirred at 45 C for
48 h and filtered through Celite . The filtrate was concentrated under reduced
pressure. The residue
was purified by column chromatography (silica gel, eluent: petroleum ether /
ethyl acetate 99:1 to
80:20). Yield: 1.44 g (27% of theory).
1H-NMR (300 MHz, CDC13): 6 [ppm] = 3.38 (s, 3H), 2.53-2.42 (m, 1H), 2.42-2.28
(m, 2H), 1.97-
1.77 (m, 2H), 1.44 (s, 9H), 0.84-0.69 (m, 1H), 0.68-0.54 (m, 1H), 0.53-0.31
(m, 2H), 0.14-0.02 (m,
1H).
Example 1.11D
tert-Butyl 2-bromo-4-cyclopropy1-4-methoxybutanoate (mixture of stereoisomers)
o-C H 3
c1C'WC H 3
Br
hCH3
0 CH3
Lithium diisopropylamide (4.9 ml, 2.0 M in tetrahydrofuran, 9.9 mmol, 1.5 eq.)
was added under
argon atmosphere at -78 C to a solution of tert-butyl 4-cyclopropy1-4-
methoxybutanoate (racemate)
(1.44 g, 6.6 mmol) in tetrahydrofuran (20 m1). The resulting mixture was
stirred at -78 C for 1 h,
followed by the dropwise addition of a solution of 1,2-dibromo-1,1,2,2-
tetrachloroethane (2.57 g,
7.9 mmol, 1.2 eq.) in tetrahydrofuran (10 ml) at -78 C. The resulting mixture
was allowed to warm
to RT, stirred further for 2 h, quenched with saturated aqueous solution of
ammonium chloride and
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extracted with ethyl acetate. The combined organic phases were washed with
water and brine, dried
over anhydrous sodium sulfate, filtered and concentrated in vacuo. The residue
was purified by
preparative thin-layer chromatography (silica gel, eluent: petroleum ether /
ethyl acetate 80:20).
Yield: 1.52 g(80% purity, 63% of theory).
1H-NMR (400 MHz, CDC13): 6 [ppm] = 4.42-4.33 (m, 1H), 3.43 / 3.36 (2s, 3H),
2.74-2.61 (m, 1H),
2.57-2.49 / 2.44-2.34 (2m, 1H), 2.29-2.19 (m, 1H), 1.47 (s, 9H), 0.87-0.72 (m,
1H), 0.71-0.60 (m,
1H), 0.53-0.36 (m, 2H), 0.19-0.03 (m, 1H).
Example 1.11E
2-Bromo-4-cyclopropy1-4-methoxybutanoic acid (mixture of stereoisomers)
o-C H3
BrC;7'
0 H
Trifluoroacetic acid (6.2 ml, 80.2 mmol, 20 eq.) was added dropwise under
argon atmosphere to an
ice-cooled solution of tert-butyl 2-bromo-4-cyclopropy1-4-methoxybutanoate
(mixture of
stereoisomers) (1.47 g, 80% purity, 4.01 mmol) in dichloromethane (40 m1). The
reaction mixture
was stirred at RT for 1.5 h, concentrated invacuo and coevaporated two times
with dichloromethane.
The residue was purified by column chromatography (silica gel, eluent:
dichloromethane / methanol
gradient). Yield: 641 mg (67% of theory).
'H-NMR (600 MHz, DMSO-d6): 6 [ppm] = 13.15 (br s, 1H), 4.44-4.34 (m, 1H), 3.32
/ 3.27 (2s, 3H),
3.04-2.97 / 2.85-2.79 / 2.72-2.62 (3m, 1H), 2.35-2.28 / 2.28-2.21 (2m, 1H),
2.19-2.11 / 2.07-2.00
(2m, 1H), 0.88-0.73 (m, 1H), 0.64-0.53 (m, 1H), 0.50-0.36 (m, 2H), 0.12-0.0
(m, 1H).
Example 1.11F
4-{[(2R,4R)-2-Bromo-4-cyclopropy1-4-methoxybutanoyllaminolbenzamide (single
stereoisomer)
C H3
)-
f H
BrrN
0 NH2
2-Bromo-4-cyclopropy1-4-methoxybutanoic acid (mixture of stereoisomers) (641
mg, 2.70 mmol),
pyridine (241 [11, 2.97 mmol, 1.1 eq.) and T3P (2.37 ml, 50% solution in ethyl
acetate, 4.06 mmol,
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1.5 eq.) were added under argon atmosphere at RT to a solution of 4-
aminobenzamide (368 mg,
2.70 mmol, 1.0 eq.) in tetrahydrofuran (15 m1). The reaction mixture was
stirred at RT for 1.5 h
before additional 4-aminobenzamide (110 mg, 0.81 mmol, 0.3 eq.) and T3P (316
IA, 50% solution
in ethyl acetate, 0.54 mmol, 0.2 eq.) were added and stirred for another 1 h.
The reaction mixture
was quenched with water and diluted with ethyl acetate. After phase
separation, the aqueous phase
was extracted with ethyl acetate. The combined organic phases were washed with
brine, dried over
anhydrous sodium sulfate, filtered and concentrated under reduced pressure.
Yield: 941 mg.
Stereoisomer separation of 941 mg
of 4-(2-bromo-4-cyclopropy1-4-methoxybutan-
amido)benzamide (mixture of stereoisomers) gave
mixture of two stereoisomers 1+2 (chiral SFC: Rt = 2.81 min): 197 mg,
single stereoisomer 3 (chiral SFC: Rt = 3.25 min, 98% ee): 134 mg,
single stereoisomer 4 (the title compound Example 1.11F) (chiral SFC: Rt =
3.58 min, 98% ee):
123 mg.
Separation method: SFC: column: Daicel OJ-H, 250 mm x 20 mm; eluent: 89%
carbon dioxide /
11% ethanol; temperature: 40 C; flow rate: 80 ml/min; UV detection: 210 nm.
Analysis method: SFC: column: Daicel OJ-3, 50 mm x 4.6 mm; eluent: 90% carbon
dioxide / 10%
methanol; temperature: 40 C; flow rate: 3 ml/min; UV detection: 210 nm.
LC-MS (method 4): R1= 1.38 min; MS (ESIneg): m/z = 353 EM-1-11-
'H-NMR (600 MHz, DM50-d6): 6 [ppm] = 10.55 (s, 1H), 7.89-7.81 (m, 3H), 7.65
(d, 2H), 7.23 (br
s, 1H), 4.74 (dd, 1H), 3.36 (s, 3H), 2.70 (dt, 1H), 2.26-2.13 (m, 2H), 0.84-
0.76 (m, 1H), 0.65-0.57
(m, 1H), 0.49-0.41 (m, 2H), 0.06-0.0 (m, 1H).
Example 1.12A
tert-Butyl 4-(difluoromethoxy)butanoate
)<H
0 F
.r() CH3
o H3
C H 3
[Bromo(difluoro)methyllftrimethyOsilane (5.28 ml, 29.6 mmol, 2.0 eq.) was
added dropwise under
argon atmosphere at RT to a solution of tert-butyl 4-hydroxybutanoate (2.50 g,
14.8 mmol) and
potassium acetate (5.82 g, 59.3 mmol, 4.0 eq.) in a mixture of dichloromethane
(8.7 ml) and water
(8.7 m1). The reaction mixture was stirred overnight and diluted with
dichloromethane and water.
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After phase separation, the aqueous phase was extracted with dichloromethane.
The combined
organic phases were dried over anhydrous sodium sulfate, filtered and
concentrated under reduced
pressure. The crude product was used without further purification. Yield: 3.82
g.
'H-NMR (600 MHz, DMSO-d6): 6 [ppm] = 6.63 (t, 1H), 3.82 (t, 2H), 2.27 (t, 2H),
1.83-1.75 (m, 2H),
1.40 (s, 9H).
Example 1.12B
tert-Butyl 2-bromo-4-(difluoromethoxy)butanoate (racemate)
H
0 F
0 C H 3
B r
hCH3
0 CH 3
A solution of n-butyllithium (12.2 ml, 1.6 M in hexane, 19.5 mmol, 1.2 eq.)
was added dropwise
under argon atmosphere at 0 C to a solution of diisopropylamine (3.2 ml, 22.8
mmol, 1.4 eq.) in
tetrahydrofuran (25 m1). The reaction mixture was stirred at 0 C for 30 min
and cooled to -78 C. A
solution of tert-butyl 4-(difluoromethoxy)butanoate (3.8 g, 90% purity, 16.3
mmol) in
tetrahydrofuran (12.5 ml) was added dropwise and stirred at -78 C for 30 min.
A solution of 1,2-
dibromo-1,1,2,2-tetrafluoroethane (5.1 g, 19.5 mmol, 1.2 eq.) was added
dropwise and stirred at -
78 C for 10 min. The reaction mixture was allowed to warm to RT, stirred for
another 1 h and
quenched with saturated aqueous solution of ammonium chloride. After phase
separation, the
aqueous phase was extracted with ethyl acetate. The combined organic phases
were washed with
brine, dried over anhydrous sodium sulfate, filtered and concentrated under
reduced pressure. The
residue was purified by column chromatography (silica gel, eluent: cyclohexane
/ ethyl acetate
gradient). Yield: 1.7 g (36% of theory).
'H-NMR (600 MHz, DMSO-d6): 6 [ppm] = 6.59 (t, 1H), 4.35-4.30 (m, 1H), 3.90-
3.80 (m, 2H), 2.30-
2.22 (m, 1H), 2.10-2.02 (m, 1H).
Example 1.13A
tert-Butyl (4R)-4-(difluoromethoxy)pentanoate (single stereoisomer)
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F
)<H
0 F
cC H3
0 H3
hC H3
o C H3
[Bromo(difluoro)methyll(trimethyl)silane (6.64 g, 32.7 mmol, 3.0 eq.) and
potassium hydrogen
difluoride (5.11 g, 65.4 mmol, 6.0 eq.) were added at RT to a mixture of tert-
butyl (4R)-4-
hydroxypentanoate (single stereoisomer) (2.00 g, 10.9 mmol) in dichloromethane
(7 ml) and water
(7 ml) in a plastic bottle. After stirring at RT for 10 h, the reaction
mixture was diluted with
dichloromethane and washed with water and brine. The organic phase was dried
over anhydrous
magnesium sulfate, filtered and evaporated to dryness under reduced pressure.
Yield: 2.17 g (85%
of theory).
1H-NMR (400 MHz, CDC13): 6 [ppm] = 6.22 (t, 1H), 4.37-4.26 (m, 1H), 2.41-2.31
(m, 2H), 1.89-
1.77 (m, 2H), 1.46 (s, 9H), 1.31 (d, 3H).
19F-NMR (376 MHz, CDC13): 6 [ppm] = -80.81 (q, 2F).
Example 1.13B
tert-Butyl (4R)-2-bromo-4-(difluoromethoxy)pentanoate (mixture of two
diastereomers)
)<H
0 F
OCH 3
Br
II hCH3
0 CH3
Lithium diisopropylamide (3.9 ml, 2.0 M in tetrahydrofuran, 7.7 mmol, 1.2 eq.)
was added at -78 C
under argon atmosphere to a solution of tert-butyl (4R)-4-(difluoromethoxy)
pentanoate (single
stereoisomer) (1.50 g, 6.4 mmol) in tetrahydrofuran (20 m1). The resulting
mixture was stirred
at -78 C for 1 h, followed by the dropwise addition of a solution of 1,2-
dibromo-1,1,2,2-
tetrachloroethane (2.30 g, 7.1 mmol, 1.1 eq.) in tetrahydrofuran (8 ml) at -78
C. The resulting
mixture was allowed to warm to RT, stirred further for 4 h, quenched with
saturated aqueous solution
of ammonium chloride and extracted with ethyl acetate. The combined organic
phases were washed
with water and brine, dried over anhydrous sodium sulfate, filtered and
evaporated to dryness in
vacuo. The crude product was purified by preparative thin layer chromatography
(silica gel, eluent:
petroleum ether! ethyl acetate 83:17). Yield: 0.85 g (85% purity, 37% of
theory).
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1H-NMR (400 MHz, CDC13): 6 [ppm] = 6.44-5.96 (m, 1H), 4.59-4.40 (m, 1H), 4.38-
4.21 (m, 1H),
2.39-2.01 (m, 2H), 1.51 /1.44 (2s, 9H), 1.36 / 1.32 (2d, 3H).
19F-NMR (376 MHz, CDC13): 6 [ppm] = -84.64 - -79.29 (m, 2F).
Example 1.14A
tert-Butyl (45)-4-(difluoromethoxy)pentanoate (single stereoisomer)
)<H
0 F
CH3
OC H3
II hC H3
0 CH3
A mixture of tert-butyl (45)-4-hydroxypentanoate (single stereoisomer) (3.00
g, 16.2 mmol, 1.0 eq.)
in dichloromethane (10 ml) and water (10 ml) was added into a plastic bottle,
followed by the
addition of [bromo(difluoro)methyllitrimethypsilane (9.86 g, 48.6 mmol, 3.0
eq.) and potassium
hydrogen difluoride (7.58 g, 97.1 mmol, 6.0 eq.) at RT. After stirring at RT
for 10 h, the reaction
mixture was diluted with dichloromethane, washed with water, dried over
anhydrous magnesium
sulfate and filtered. The filtrate was concentrated under reduced pressure and
used in the subsequent
reaction without further purification. Yield: 3.50 g (92% of theory).
1H-NMR (400 MHz, CDC13): 6 [ppm] = 6.47-5.97 (m, 1H), 4.39-4.18 (m, 1H), 2.39-
2.25 (m, 2H),
1.90-1.72 (m, 2H), 1.45 (s, 9H), 1.30 (d, 3H).
Example 1.14B
tert-Butyl (45)-2-bromo-4-(difluoromethoxy)pentanoate (mixture of two
diastereomers)
)<H
0 F
OCH 3
Br
II hCH3
0 CH3
Lithium diisopropylamide (8.9 ml, 2.0 M in tetrahydrofuran, 17.8 mmol, 1.2
eq.) was added at -78 C
to a solution of tert-butyl (45)-4-(difluoromethoxy)pentanoate (single
stereoisomer) (3.50 g,
14.8 mmol, 1.0 eq.) in tetrahydrofuran (40 m1). After stirring at -78 C for 1
h, 1,2-dibromo-1,1,2,2-
tetrachloroethane (5.79 g, 17.8 mmol, 1.2 eq.) in tetrahydrofuran (15 ml) was
added. The resulting
mixture was allowed to warm to RT and stirred for further 4 h. The reaction
mixture was quenched
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with saturated aqueous solution of ammonium chloride and extracted with ethyl
acetate. The
combined organic layers were washed with water and brine, dried over anhydrous
sodium sulfate,
filtered and concentrated under reduced pressure. The crude product was
purified preparative thin-
layer chromatography (silica gel, eluent: petroleum ether! ethyl acetate
91:9). Yield: 2.09 g (45% of
theory).
1H-NMR (400 MHz, CDC13): 6 [ppm] = 6.48-5.92 (m, 1H), 4.52-4.44 (m, 1H), 4.36-
4.22 (m, 1H),
2.44-1.98 (m, 2H), 1.49! 1.49 (2s, 9H), 1.36! 1.32 (2d, 3H).
Example 1.15A
Cyclobutyl 4-methylbenzene-1-sulfonate
(R.
H 3C 1IS-0
0
A solution of cyclobutanol (13 ml, 170 mmol, 1.0 eq.) in pyridine (86 ml) was
cooled to 0 C, then
4-methylbenzene-1-sulfonyl chloride (34.9 g, 183 mmol, 1.1. eq.) was added in
portions and stirring
was continued for 80 min. Subsequently, the reaction mixture was poured to a
mixture of ice and
water, extracted with dichloromethane, washed with saturated aqueous solution
of sodium
bicarbonate and water, dried over anhydrous sodium sulfate, filtered and
concentrated under reduced
pressure. The residue was then taken up in n-heptane and concentrated again
for three times. The
crude product was used in the subsequent step without further purification.
Yield: 33.9 g (90% of
theory).
GC-MS (method 6): Rt = 6.65 min; MS (APCIpos): m/z = 227 [M+I-11+
Example 1.15B
4-(Cyclobutyloxy)butan-1-ol
1:117
0
H
A solution of butane-1,4-diol (39.8 g, 442 mmol, 10.0 eq.) and potassium
hydroxide (4.96 g, 88.4
mmol, 2.0 eq.) in 1,4-dioxane (220 ml) was heated to 105 C before a solution
of cyclobutyl 4-
methylbenzene-l-sulfonate (10.0 g, 44.2 mmol, 1.0 eq.) in 1,4-dioxane (46 ml)
was added. The
mixture was stirred at 105 C for 4 h, then diatomaceous earth was added and
the suspension was
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concentrated under reduced pressure. The residue was purified by column
chromatography (silica
gel, eluent: cyclohexane / ethyl acetate 1:1 to 3:7 to 1:9). Yield: 7.1 g (71%
purity, 79% of theory).
GC-MS (method 6): Rt = 3.61 min; MS (APCIpos): m/z = 145 [WHY'
Example 1.15C
4-(Cyclobutyloxy)butanoic acid
0
H
0
To a solution of chromium trioxide (11.1 g, 111 mmol, 8.0 eq.) in water (30
ml) at 0 C was added
sulfuric acid (9.7 ml) and water (60 m1). The resulting mixture was stirred at
0 C for 15 min and then
transferred to a solution of 4-(cyclobutyloxy)butan-1-ol (2.00 g, 13.9 mmol,
1.0 eq.) in acetone
(40 ml) at RT. Stirring was continued at RT for 16 h, followed by the addition
of water and extraction
with dichloromethane. The combined organic phases were washed with brine,
dried over anhydrous
sodium sulfate, filtered and concentrated under reduced pressure. The crude
product was used in the
subsequent step without further purification. Yield: 1.30 g of crude product.
GC-MS (method 6): Rt = 4.05 min; MS (APCIpos): m/z = 159 [M+I-11+
Example 1.15D
Ethyl 4-(cyclobutyloxy)butanoate
X-117
OC
0
H3
To a crude mixture of 4-(cyclobutyloxy)butanoic acid (1.30 g, 8.22 mmol, 1.0
eq.) in ethanol (29 ml)
at 0 C was added dropwise thionyl chloride (1.2 ml, 16 mmol, 2.0 eq.). The
resulting mixture was
allowed to warm to RT and stirring was continued for 4 h before the mixture
was concentrated under
reduced pressure and coevaporated with toluene. The residue was purified by
column
chromatography (silica gel, eluent: cyclohexane / ethyl acetate 9:1 to 4:1 to
7:3 to 3:1 to 1:1). Yield:
100 mg (76% purity, 5% of theory) and 270 mg (86% purity, 15% of theory).
GC-MS (method 6): R1= 4.15 min; MS (APCIpos): m/z = 187 [M+I-11+
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Example 1.15E
Ethyl 2-bromo-4-(cyclobutyloxy)butanoate (racemate)
0
H 3
Br
0
To a solution of diisopropylamine (280 IA, 2.0 mmol, 1.4 eq.) in
tetrahydrofuran (4.6 ml) was added
a solution of n-butyllithium (700 IA, 2.5 M in hexane, 1.7 mmol, 1.2 eq.) at 0
C. After stirring at this
temperature for 30 min, the mixture was cooled to -78 C and a solution of 1,2-
dibromo-1,1,2,2-
tetrachloroethane (566 mg, 1.74 mmol, 1.2 eq.) in tetrahydrofuran (4.6 ml) was
added dropwise. The
mixture was stirred at -78 C for 45 min, followed by the addition of further
1,2-dibromo-1,1,2,2-
tetrachloroethane (566 mg, 1.74 mmol, 1.2 eq.) in portions. The resulting
mixture was allowed to
warm to RT over a period of 1 h, quenched by the addition of saturated aqueous
solution of
ammonium chloride and extracted with dichloromethane. The combined organic
layers were dried
over anhydrous sodium sulfate, filtered and concentrated under reduced
pressure. The residue was
purified by column chromatography (silica gel, eluent: cyclohexane / ethyl
acetate 98:2 to 96:4 to
94:6 to 92:8 to 9:1). Yield: 120 mg (75% purity, 23% of theory).
GC-MS (method 6): R1= 5.13 min; MS (APCIpos): m/z = 266 [M+I-11+
Example 1.16A
Ethyl 5,5,5-trifluoro-4-hydroxypentanoate (racemate)
0
F>l'OC H3
0 H
To a solution of ethyl 5,5,5-trifluoro-4-oxopentanoate (4.00 g, 20.2 mmol, 1.0
eq.) in ethanol (40 ml)
was added sodium borohydride (3.82 g, 100.9 mmol, 5.0 eq.) in portions at 0 C.
After stirring at RT
for 5 h, the reaction mixture was poured into a mixture of aqueous
hydrochloric acid (0.5 N) and
ethyl acetate and stirred for 10 min. The aqueous solution was extracted with
ethyl acetate, the
combined organic layers were washed with water and brine, dried over anhydrous
sodium sulfate,
filtered and concentrated under reduced pressure. The residue was purified by
column
chromatography (silica gel, eluent: petroleum ether / ethyl acetate 92:8).
Yield: 1.65 g (39% of
theory).
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1H-NMR (400 MHz, DMSO-d6): 6 [ppm] = 6.22 (d, 1H), 4.06 (q, 2H), 4.01-3.89 (m,
1H), 2.48-2.41
(m, 2H), 1.90-1.79 (m, 1H), 1.73-1.60 (m, 1H), 1.18 (t, 3H).
Example 1.16B
Ethyl 4-{ [tert-butyl(dimethypsilyll oxy} -5,5,5 -trifluoropentanoate
(racemate)
0
CH3
0 /L' 113
/SiC H3
H3C fl'CH
CH3
To a solution of ethyl 5,5,5-trifluoro-4-hydroxypentanoate (racemate) (1.10 g,
5.3 mmol, 1.0 eq.) in
acetonitrile (15 ml) were added imidazole (1.40 g, 21.1 mmol, 4.0 eq.) and
tert-butyldimethylsilyl
chloride (2.40 g, 15.8 mmol, 3.0 eq.) at RT. The mixture was then heated to
reflux and stirred at this
temperature for 7 h. After concentrating under reduced pressure, the residue
was dissolved in ethyl
acetate and washed with water and brine. The organic phase was then dried over
anhydrous sodium
sulfate, filtered and concentrated under reduced pressure. The residue was
purified by column
chromatography (silica gel, eluent: petroleum ether). Yield: 1.30 g (75% of
theory).
1H-NMR (400 MHz, CDC13): 6 [ppm] = 4.16 (q, 2H), 4.11-4.01 (m, 1H), 2.55-2.37
(m, 2H), 2.09-
1.84 (m, 2H), 1.27 (t, 3H), 0.91 (s, 9H), 0.11 (s, 3H), 0.10 (s, 3H).
Example 1.16C
Ethyl 2-bromo-4-{[tert-butyl(dimethypsilylloxy}-5,5,5-trifluoropentanoate
(mixture of
stereoisomers)
0
F>ly.Y.00H3
H3C
H3C:0 Br
Si"
H3C1 µCH3
CH3
To a solution of ethyl 4-{[tert-butyl(dimethyl)silylloxy} -5,5,5-
trifluoropentanoate (racemate)
(1.30 g, 4.0 mmol, 1.0 eq.) in tetrahydrofuran (20 ml) was added a solution of
lithium
diisopropylamide (2.0 ml, 2.0 M in tetrahydrofuran, 4.0 mmol, 1.0 eq.) at -78
C. After stirring at -
78 C for 1 h, a solution of 1,2-dibromo-1,1,2,2-tetrachloroethane (1.42 g, 4.4
mmol, 1.1 eq.) in
tetrahydrofuran (6 ml) was added. The resulting mixture was allowed to warm to
RT and stirred
further for 4 h. Water was then added and the mixture was extracted with ethyl
acetate. The combined
organic layers were washed with water and brine, dried over anhydrous sodium
sulfate, filtered and
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concentrated under reduced pressure. The residue was purified by column
chromatography (silica
gel, eluent: petroleum ether! ethyl acetate 98:2). Yield: 1.10 g (69% purity,
49% of theory).
1H-NMR (300 MHz, CDC13): 6 [ppm] = 4.39-4.04 (m, 3H), 2.72-2.41 (m, 1H), 2.37-
2.21 (m, 1H),
2.13-1.79 (m, 1H), 1.45-1.17 (m, 3H), 0.94-0.88 (m, 9H), 0.23-0.00 (m, 6H).
Example 1.17A
Ethyl 5,5,5-trifluoro-4-methoxypentanoate (racemate)
0
H3
0
H
To a solution of ethyl 5,5,5-trifluoro-4-hydroxypentanoate (racemate) (3.0 g,
14.4 mmol, 1.0 eq.) in
1,2-dichloroethane (40 ml) were added freshly prepared silver(I) oxide (10.0
g, 43.2 mmol, 3.0 eq.)
and iodomethane (9.0 ml, 140 mmol, 10.0 eq.) at RT and the mixture was heated
to 35 C. After
stirring for 54 h at this temperature, the reaction mixture was cooled to RT,
filtered through Celite
and concentrated under reduced pressure. The residue was dissolved in ethyl
acetate, washed with
water, dried over anhydrous sodium sulfate, filtered and concentrated under
reduced pressure. The
crude product was purified by column chromatography (silica gel, eluent:
petroleum ether! ethyl
acetate 96:4). Yield: 600 mg (20% of theory).
1H-NMR (300 MHz, DMSO-d6): 6 [ppm] = 4.07 (q, 2H), 3.97-3.80 (m, 1H), 3.45 (s,
3H), 2.48-2.40
(m, 2H), 1.96-1.82 (m, 1H), 1.81-1.62 (m, 1H), 1.19 (t, 3H).
Example 1.17B
Ethyl 2-bromo-5,5,5-trifluoro-4-methoxypentanoate (mixture of stereoisomers)
0
F>IL
OC H3
0 Br
H3C'
To a solution of ethyl 5,5,5-trifluoro-4-methoxypentanoate (racemate) (2.20 g,
10.1 mmol, 1.0 eq.)
in tetrahydrofuran (40 ml) was added a solution of lithium diisopropylamide
(6.0 ml, 2.0 M in
tetrahydrofuran, 12.1 mmol, 1.0 eq.) at -78 C. After stirring at -78 C for 1
h, 1,2-dibromo-1,1,2,2-
tetrachloroethane (3.93 g, 12.1 mmol, 1.2 eq.) in tetrahydrofuran (15 ml) was
added, the resulting
mixture was allowed to warm to RT and stirring was continued for 2 h. The
reaction mixture was
treated with saturated aqueous solution of ammonium chloride and extracted
with ethyl acetate. The
combined organic layers were washed with water and brine, dried over anhydrous
sodium sulfate,
filtered and concentrated under reduced pressure. The crude product was
purified by preparative thin-
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layer chromatography (silica gel, eluent: petroleum ether / ethyl acetate
91:9). Yield: 1.46 g (84%
purity, 42% of theory).
1H-NMR (400 MHz, CDC13): 6 [ppm] = 4.55-4.42 (m, 1H), 4.34-4.21 (m, 2H), 4.00-
3.74 (m, 1H),
3.63 (d, 2H), 3.51 (s, 1H), 2.38 (s, 1H), 2.34-2.22 (m, 1H), 1.41-1.24 (m,
3H). Additional signals of
minor diastereomer visible.
Example 1.17C
2-Bromo-5,5,5-trifluoro-4-methoxypentanoic acid (mixture of stereoisomers)
0
H
0 Br
H3C'
General Method 10 was carried out with ethyl 2-bromo-5,5,5-trifluoro-4-
methoxypentanoate
.. (mixture of stereoisomers) (1.26 g, 84% purity, 3.61 mmol, 1.0 eq.) and
lithium hydroxide (432 mg,
18.1 mmol, 5.0 eq.) in a mixture of tetrahydrofuran / water (3.6:1, 115 m1).
The crude mixture was
used in the subsequent reaction without further purification. Yield: 1.10 g
(85% purity, 99% of
theory).
GC-MS (method 6): R1= 3.51 /3.56 min; MS (APCIpos): m/z = 265 [M+I-11+
Example 1.17D
4-{ [(2R,4R)-2-B romo -5 ,5 ,5 -trifluoro-4-methoxypentanoyl] amino }
benzamide (single stere oi some r)
0
N H 2
H3C' r
F . N
H
0 B
General Method 11 was carried out with 4-aminobenzamide (518 mg, 3.81 mmol,
1.0 eq.), 2-bromo-
5,5,5-trifluoro-4-methoxypentanoic acid (mixture of stereoisomers) (1.13 g,
85% purity, 4.19 mmol,
0.95 eq.), T3P (3.4 ml, 50% solution in ethyl acetate, 5.71 mmol, 1.5 eq.) and
pyridine (339 [11,
4.19 mmol, 1.1 eq.) in tetrahydrofuran (15 ml) for 1 h including the following
variations of the
procedure: Iced water was added to the reaction mixture which was then
extracted with
dichloromethane and concentrated under reduced pressure. The crude mixture was
purified by
column chromatography (silica gel, eluent: dichloromethane / methanol 98:2 to
80:20). Yield: 910
mg (91% purity, 57% of theory).
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Stereoisomer separation of 910 mg of 4-[(2-bromo-5,5,5-trifluoro-4-
methoxypentanoyl)aminol-
benzamide (mixture of stereoisomers) gave:
single stereoisomer 1 (the title compound 1.17D) (chiral SFC: R1= 2.44 min,
99% de): 53.1 mg (4%
of theory),
single stereoisomer 2 (chiral SFC: Rt = 2.72 min): 46.6 mg,
single stereoisomer 3 (chiral SFC: Rt = 3.46 min): 215 mg,
single stereoisomer 4 (chiral SFC: Rt = 7.83 min): 259 mg.
Separation method 1: SFC: column: Daicel Chiralpak IE 5 um, 250 mm x 20 mm;
eluent: 80%
carbon dioxide /20% methanol; temperature: 35 C; flow rate: 80 ml/min; UV
detection: 210 nm.
Separation method 2: Single stereoisomer 1 and single stereoisomer 2 eluted as
a mixture in the first
separation, this mixture was then separated according to the following
conditions: SFC: column:
Daicel Chiralpak IF 5 um, 250 mm x 20 mm; eluent: 88% carbon dioxide / 12%
methanol;
temperature: 35 C; flow rate: 80 ml/min; UV detection: 210 nm.
Analysis method: SFC: column: Daicel Chiralpak 1E-3 3 um, 250 mm x 4.6 mm;
eluent: 90% carbon
dioxide / 10% methanol; flow rate: 3.0 ml/min; UV detection: 210 nm.
LC-MS (method 5): R1= 1.04 min; MS (ESIpos): m/z = 383 [M+Hl+
1H-NMR (400 MHz, DM50-d6): 6 [ppm] = 10.73 (s, 1H), 7.92-7.83 (m, 3H), 7.67
(d, 2H), 7.27 (br
s, 1H), 4.72 (dd, 1H), 3.90-3.82 (m, 1H), 3.44 (s, 3H), 2.64-2.56 (m, 1H),
2.22-2.14 (m, 1H).
Example 1.18A
Ethyl 5,5-difluoro-4-oxopentanoate
0 F
________________________________________ H
0 C H3
0
[Bromo(difluoro)methyll(trimethyl)silane (74.04 g, 364.5 mmol, 1.5 eq.),
triphenylphosphine (70.12
g, 267.3 mmol, 1.1 eq.) and 1,3-dimethy1-3,4,5,6-tetrahydro-2(1H)-pyrimidinone
(62.30 g, 486.1
mmol, 2.0 eq.) were added under argon atmosphere at RT to a solution of ethyl
4-chloro-4-
oxobutanoate (40.0 g, 243.0 mmol, 1.0 eq.) in acetonitrile (240 m1). After
stirring for 5 h at RT, the
resulting mixture was quenched with water (200 ml) and pyridine (76.90 g,
972.1 mmol, 4.0 eq.).
After stirring for further 1.5 h at 80 C, the reaction mixture was cooled to
RT and concentrated under
reduced pressure. The residue was diluted with water and extracted with methyl
tert-butyl ether. The
combined organic phases were washed with water and brine, dried over anhydrous
sodium sulfate,
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filtered and concentrated under reduced pressure. The residue was purified by
column
chromatography (silica, gel, eluent: petroleum ether! ethyl acetate 95:5).
Yield: 15.20 g (70% purity,
22% of theory).
1H-NMR (400 MHz, CDC13): 6 [ppm] = 5.94-5.60 (m, 1H), 4.23-4.07 (m, 2H), 2.98-
2.94 (m, 1H),
2.71-2.64 (m, 2H), 2.14-2.06 (m, 1H), 1.30-1.22 (m, 3H).
19F-NMR (376 MHz, CDC13): 6 [ppm] = -127.60 (s, 2F).
Example 1.18B
Ethyl 5,5-difluoro-4-hydroxypentanoate (racemate)
OH F
cH
OCH 3
0
Sodium borohydride (2.23 g, 59.1 mmol, 1.0 eq.) was added at 0 C to a solution
of ethyl 5,5-difluoro-
4-oxopentanoate (15.20 g, 70% purity, 59.1 mmol) in tetrahydrofuran (140 ml)
and ethanol (20 m1).
After stirring at 0 C for 2 h, the reaction mixture was quenched with aqueous
hydrochloric acid (3
N) at 0 C and concentrated under reduced pressure. The residue was dissolved
in ethyl acetate and
washed with water and brine. The organic phase was dried over anhydrous sodium
sulfate, filtered
and concentrated under reduced pressure. The residue was purified by column
chromatography
(silica gel, eluent: petroleum ether! ethyl acetate 90:10). Yield: 5.40 g (48%
of theory).
1H-NMR (400 MHz, DMSO-d6): 6 [ppm] = 5.96-5.67 (m, 1H), 5.53 (d, 1H), 4.08-
4.00 (m, 2H), 3.69-
3.53 (m, 1H), 2.48-2.32 (m, 2H), 1.82-1.69 (m, 1H), 1.64-1.51 (m, 1H), 1.18
(t, 3H).
19F-NMR (376 MHz, DMSO-d6): 6 [ppm] = -128.80 (q, 2F).
Example 1.18C
Ethyl 5,5-difluoro-4-methoxypentanoate (racemate)
H3C,
0 F
_________________________________________ H
OCH 3
0
Iodomethane (40.39 g, 284.6 mmol, 10.0 eq.) was added under argon atmosphere
at RT to a mixture
of ethyl 5,5-difluoro-4-hydroxypentanoate (racemate) (5.40 g, 28.5 mmol, 1.0
eq.) and freshly
prepared silver(I) oxide (19.78 g, 85.4 mmol, 3.0 eq.) in dichloroethane (80
m1). After stirring at
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45 C for 72 h, the resulting mixture was filtered through Celite . The
filtrate was concentrated under
reduced pressure. The residue was purified by column chromatography (silica
gel, eluent: petroleum
ether! ethyl acetate 95:5). Yield: 2.40 g (90% purity, 39% of theory).
1H-NMR (300 MHz, CDC13): 6 [ppm] = 5.84-5.45 (m, 1H), 4.17-4.09 (m, 2H), 3.47
(s, 3H), 3.42-
3.36 (m, 1H), 2.54-2.40 (m, 2H), 2.03-1.89 (m, 1H), 1.88-1.74 (m, 1H), 1.28
(t, 3H).
19F-NMR (282 MHz, CDC13): 6 [ppm] = -126.70 (d, 2F).
Example 1.18D
Ethyl 2-bromo-5,5-difluoro-4-methoxypentanoate (mixture of stereoisomers)
H3C,
0 F
( H
0 C H3
B r
0
Lithium diisopropylamide solution (8.0 ml, 2.0 M in tetrahydrofuran, 16.0
mmol, 1.2 eq.) was added
under argon atmosphere at -78 C to a solution of ethyl 5, 5-difluoro-4-
methoxypentanoate (racemate)
(2.90 g, 90% purity, 13.3 mmol, 1.0 eq.) in tetrahydrofuran (70 m1). The
resulting mixture was stirred
at -78 C for 1 h, followed by the addition of a solution of 1,2-dibromo-
1,1,2,2-tetrachloroethane
(5.20 g, 16.0 mmol, 1.2 eq.) in tetrahydrofuran (20 ml) dropwise at the same
temperature. After
warming to RT and stirring for further 2 h, the reaction mixture was quenched
with saturated aqueous
solution of ammonium chloride and extracted with ethyl acetate. The combined
organic phases were
washed with water and brine, dried over anhydrous sodium sulfate, filtered and
concentrated under
reduced pressure. The residue was purified by preparative thin-layer
chromatography (silica gel,
eluent: petroleum ether! ethyl acetate 80:20). Yield: 1.85 g (85% purity, 43%
of theory).
1H-NMR (400 MHz, CDC13): 6 [ppm] = 5.87-5.70 (m, 1H), 4.50-4.22 (m, 3H), 3.66-
3.42 (m, 4H),
2.23-2.16 (m, 2H), 1.35-1.26 (m, 3H).
19F-NMR (376 MHz, CDC13): 6 [ppm] = -126.60 (s, 2F).
Example 1.18E
2-Bromo-5,5-difluoro-4-methoxypentanoic acid (mixture of stereoisomers)
H 3C, 0 F
H
OH
B r
0
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Lithium hydroxide (255 mg, 10.6 mmol, 2.0 eq.) was added at RT to a solution
of ethyl 2-bromo-
5,5-difluoro-4-methoxypentanoate (mixture of stereoisomers) (1.72 g, 85%
purity, 5.3 mmol) in a
mixture of tetrahydrofuran and water (3:1, 20 m1). The reaction mixture was
stirred at RT for 100 min
and then acidified with aqueous hydrochloric acid (1 N). After removing all
volatiles under reduced
.. pressure, the resulting mixture was extracted with ethyl acetate. The
combined organic phases were
washed with brine, dried over anhydrous sodium sulfate, filtered and
concentrated under reduced
pressure (at <30 C and >100 mbar). The crude product was used without further
purification. Yield:
1.65 g.
Example 1.18F
4-{[(2R,4R)-2-Bromo-5,5-difluoro-4-methoxypentanoyllaminolbenzamide (single
stereoisomer)
H3C,
0 F
______________________________________ H
H N
Br "'fl'
0 N H 2
0
2-Bromo-5,5-difluoro-4-methoxypentanoic acid (mixture of stereoisomers) (1.65
g, 90% assumed
purity of crude material, 6.0 mmol), pyridine (0.53 ml, 6.6 mmol, 1.1 eq.) and
T3P (5.3 ml, 50%
solution in ethyl acetate, 9.0 mmol, 1.5 eq.) were added under argon
atmosphere at RT to a mixture
of 4-aminobenzamide (817 mg, 6.0 mmol, 1.0 eq.) in tetrahydrofuran (20 m1).
The reaction mixture
was stirred at RT for 1 h, mixed with water, stirred for additional 15 min and
mixed with additional
water. After addition of ethyl acetate and phase separation, the aqueous phase
was extracted with
ethyl acetate. The combined organic phases were washed with brine, dried over
anhydrous sodium
sulfate, filtered and concentrated under reduced pressure. The residue was
crystallized with
dichloromethane, filtered and dried in vacuo. Yield: 993 mg (94% purity, 43%
of theory). The
combined mother liquids were concentrated under reduced pressure. The residue
was purified by
column chromatography (silica gel, eluent: dichloromethane / methanol
gradient). Yield: 297 mg
(13% of theory).
LC-MS (method 4): R1= 1.26 min; MS (ESIpos): m/z = 365 [M+1-11+
.. Stereoisomer separation of 1410 mg of 4-[(2-bromo-5,5-difluoro-4-
methoxypentanoyl)aminol-
benzamide (mixture of stereoisomers) gave
mixture of two stereoisomers 1+2 (chiral SFC 1: Rt = 1.34 / 1.45 min): 290 mg,
single stereoisomer 3 (the title compound Example 1.18F) (chiral SFC 1: Rt =
1.18 min, >98% ee):
330 mg,
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single stereoisomer: (chiral SFC 2: Rt = 1.40 min): 410 mg.
Separation method: SFC: column: Chiralpak AD-H 5 um, 250 mm x 20 mm; eluent:
carbon dioxide
/ ethanol, gradient: 0 min 80% carbon dioxide, 5.00-8.43 min 70% carbon
dioxide, 8.53-11.31 min
80% carbon dioxide; temperature: 40 C; flow rate: 100 ml/min; UV detection:
210 nm.
Analysis method: SFC 1: column: Chiralpak AD-H, 50 mm x 4.6 mm; eluent: 80%
carbon dioxide /
20% ethanol; temperature: 40 C; flow rate: 3 ml/min; UV detection: 210 nm. SFC
2: column:
Chiralpak AD-H, 50 mm x 4.6 mm; eluent: 60% carbon dioxide / 40% ethanol;
temperature: 40 C;
flow rate: 3 ml/min; UV detection: 210 nm.
'H-NMR (600 MHz, DM50-d6): 6 [ppm] = 10.61 (s, 1H), 7.90-7.81 (m, 3H), 7.65
(d, 2H), 7.24 (br
s, 1H), 6.17 (dt, 1H), 4.76 (dd, 1H), 3.71-3.62 (m, 1H), 3.49 (s, 3H), 2.21-
2.11 (m, 2H).
Example 1.19A
4-{[(2R)-2-Bromopropanoyllaminol-2-fluorobenzamide (single stereoisomer)
C H 3
= H
BrrN
0 N H 2
0
(2R)-2-Bromopropanoic acid (single stereoisomer) (4.0 g, 26.1 mmol), pyridine
(2.3 ml, 28.8 mmol,
1.1 eq.) and T3P (22.9 ml, 50% solution in ethyl acetate, 39.2 mmol, 1.5 eq.)
were added under argon
atmosphere at RT to a mixture of 4-amino-2-fluorobenzamide (4.0 g, 26.1 mmol,
1.0 eq.) in
tetrahydrofuran (60 m1). The reaction mixture was stirred at RT for 1 h, mixed
with water and stirred
for additional 1 h. The forming precipitate was filtered, washed with water
and dried in vacuo . Yield:
5.7 g (75% of theory). The combined filtrates were extracted with ethyl
acetate. The combined
organic phases were washed with brine, dried over anhydrous sodium sulfate,
filtered and
concentrated under reduced pressure. The residue was crystallized from water
and the precipitate
dried in vacuo . Yield: 1.5 g (20% of theory).
main batch: LC-MS (method 1): Rt = 0.61 min; MS (ESIpos): m/z = 289 [M+I-11+
main batch: 'H-NMR (600 MHz, DM50-d6): 6 [ppm] = 10.68 (s, 1H), 7.70 (t, 1H),
7.64 (dd, 1H),
7.57-7.46 (m, 2H), 7.36 (dd, 1H), 4.69 (q, 1H), 1.76 (d, 3H).
Example 1.20A
4-{[(2R)-2-Bromopropanoyllaminolbenzamide (single stereoisomer)
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C H3
z H
Br N
-r
0 J NH 2
(2R)-2-Bromopropanoic acid (single stereoisomer) (1.00 g, 6.54 mmol), pyridine
(0.58 ml,
7.19 mmol, 1.1 eq.) and T3P (5.73 ml, 50% solution in ethyl acetate, 9.81
mmol, 1.5 eq.) were added
under argon atmosphere at RT to a mixture of 4-aminobenzamide (890 mg, 6.54
mmol, 1.0 eq.) in
tetrahydrofuran (15 m1). The reaction mixture was stirred at RT for 30 min.
After removing all
volatiles under reduced pressure, the residue was mixed with additional water.
The forming
precipitate was filtered, washed with water and dried in vacuo . Yield: 1.77 g
(quantitative of theory).
LC-MS (method 15): Rt = 0.97 min; MS (ESIpos): m/z = 271 [M+I-11+
'H-NMR (600 MHz, DM50-d6): 6 [ppm] = 10.52 (s, 1H), 7.87 (br s, 1H), 7.86 (d,
2H), 7.66 (d, 2H),
7.26 (br s, 1H), 4.71 (q, 1H), 1.76 (d, 3H).
Example 1.21A
Trimethyl R5R)-5-methy1-4,5-dihydrofuran-2-yll oxy} si lane (single
stereoisomer)
C H 3
.40
C
H3
C H3
Under argon atmosphere, n-butyllithium solution (3.34 1, 1.6 M in hexanes,
5.35 mol, 1.1 eq.) was
added at 0 C within 1.5 h to a solution of diisopropylethylamine (818 ml, 5.84
mol, 1.2 eq.) in
tetrahydrofuran (5.0 1) and stirred for 30 min at 0 C before cooled to -78 C.
A solution of (5R)-5-
methyldihydrofuran-2(3H)-one (single stereoisomer) (487 g, 4.86 mol) in
tetrahydrofuran (500 ml)
was added at -78 C within 1 h and the reaction mixture stirred at -78 C for 90
min, followed by the
dropwise addition of chloro(trimethyl)silane (803 ml, 6.32 mol, 1.3 eq.). The
reaction mixture was
stirred at -78 C for 1 h and allowed to warm to RT overnight while stirring
was continued. The
suspension was decanted and the filtrate evaporated under reduced pressure and
again suspended in
n-pentane. The mixture was stirred for 30 min at -10 C and filtered. The
filtrate was evaporated
under reduced pressure. The crude material was used without further
purification. Yield: 759 g (85%
purity, 79% of theory).
1H-NMR (600 MHz, CDC13): 6 [ppm] = 4.53-4.46 (m, 1H), 3.45 (t, 1H), 2.63 (ddd,
1H), 2.10 (ddd,
1H), 1.20 (d, 3H), 0.12 (s, 9H).
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Example 1.21B
(5R)-3-Bromo-5-methyldihydrofuran-2(3H)-one (mixture of two diastereomers)
C H 3
Br4
0
Bromine (197 ml, 3.83 mol, 1.0 eq.) was added slowly at -78 C to a solution of
trimethyl {[(5R)-5-
methyl-4,5-dihydrofuran-2-ylloxy}silane (single stereoisomer) (759 g, 87%
purity, 3.83 mol) in
dichloromethane (7.6 1). The reaction mixture was allowed to warm to RT
overnight while stirring
was continued and then concentrated under reduced pressure. The residue was
purified with a thin-
film vacuum evaporator (dosing of crude material: 50 ml/min, 107 C, 0.5 mbar).
Yield: 563 g
(translcis: about 2:1, about 80% purity, about 66% of theory).
1H-NMR (600 MHz, CDC13): 6 [ppm] = 4.80-4.67 (m, 1H, trans-diastereomer), 4.59-
4.47 (m, 2H,
cis-diastereomer), 4.35 (dd, 1H, trans-diastereomer), 2.93 (ddd, 1H, cis-
diastereomer), 2.46 (ddd,
1H, trans-diastereomer), 2.42-2.35 (m, 1H, cis-diastereomer), 2.30-2.20 (m,
1H, trans-
diastereomer), 2.15-2.04 (m, 1H, cis-diastereomer), 1.38 (d, 3H, cis-
diastereomer), 1.34 (d, 3H,
trans-diastereomer).
Example 1.21C
Methyl (45)-2-bromo-4-methoxypentanoate (mixture of two diastereomers)
CYCH3
4CH3
Br CH3
0
Trimethoxymethane (688 ml, 6.28 mol, 2.5 eq.) followed by sulfuric acid (16.7
ml, 314 mmol,
0.125 eq.) were added at RT to a solution of (5R)-3-bromo-5-methyldihydrofuran-
2(3H)-one
(mixture of two diastereomers) (563 g, about 80% purity, 2.51 mol) in methanol
(5.11). The reaction
mixture was stirred at 50 C for 24 h and concentrated under reduced pressure.
Afterwards,
trimethoxymethane (688 ml, 6.28 mol, 2.5 eq.) followed by sulfuric acid (16.7
ml, 314 mmol,
0.125 eq.) were again added at RT to a solution of the crude material in
methanol (5.1 1). The reaction
mixture was stirred at 50 C for 24 h and concentrated under reduced pressure.
This procedure was
repeated one more time to allow for complete conversion of the reaction. The
resulting residue was
mixed with ethyl acetate and the mixture washed with 0.5 N aqueous sodium
hydroxide solution.
The organic phase was dried over sodium sulfate, filtered and concentrated
under reduced pressure.
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The crude material was used without further purification. Yield: 625 g (94%
purity, quantitative of
theory).
1H-NMR (400 MHz, DMSO-d6): 6 [ppm] = 4.55-4.43 (m, 1H, two diastereomers),
3.68 (s, 3H, minor
diastereomer), 3.67 (s, 3H, major diastereomer), 3.46-3.37 (m, 1H, major
diastereomer), 3.34-3.25
(m, 1H, minor diastereomer), 3.19 (s, 3H, major diastereomer), 3.15 (s, 3H,
minor diastereomer),
2.22-2.05 (m, 1H, major diastereomer), 1.99-1.90 (m, 1H, minor diastereomer),
1.11-1.05 (m, 3H,
two diastereomers).
Example 2.1A
3 -(2-Bromo -4 -chloropheny1)-2 -methylprop-2-enal (E/Z mixture)
CI Br
CH3
0
Propionaldehyde (3.6 ml, 50.1 mmol, 1.1 eq.) was added dropwise at RT to a
mixture of 2-bromo-4-
chlorobenzaldehyde (10.0 g, 45.6 mmol) and sodium hydroxide (0.18 g, 4.6 mmol,
0.1 eq.) in
methanol (100 m1). The mixture was stirred at RT for 1 h, followed by the
addition of acetic acid to
adjust the pH value to 6. The resulting mixture was stirred further 16 h and
then concentrated under
reduced pressure. The residue was diluted with water and extracted with ethyl
acetate. The combined
organic layers were washed with water and brine, dried over anhydrous sodium
sulfate, filtered and
concentrated under reduced pressure. The residue was purified by column
chromatography (silica
gel, eluent: petroleum ether! ethyl acetate 10:1). Yield: 6.0 g (50% of
theory).
LC-MS (method 10): Rt = 1.28 min; MS (ESIpos): m/z = 259 [M+I-11+
1H-NMR (400 MHz, DM50-d6): 6 [ppm] = 9.70 (s, 1H), 7.93-7.91 (m, 1H), 7.59-
7.56 (m, 2H), 7.52-
7.50 (m, 1H), 1.81! 1.81 (2s, 3H).
Example 2.1B
3 -(2-Bromo -4 -chloropheny1)-2 -methylprop-2-en-1 -ol (E/Z mixture)
CI Br
C H3
OH
A mixture of 3-(2-bromo-4-chloropheny1)-2-methylprop-2-enal (E/Z mixture)
(10.0 g, 38.5 mmol,
1.0 eq.) in tetrahydrofuran (90 ml) was added at 0 C to a suspension of sodium
borohydride (7.29 g,
192.7 mmol, 5.0 eq.) in tetrahydrofuran (10 ml) and methanol (10 m1). The
reaction mixture was
stirred at RT for 3 h, followed by addition of aqueous hydrochloric acid to
adjust the pH value to 6.
The volatiles were removed under reduced pressure and the aqueous phase was
extracted with ethyl
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acetate. The combined organic layers were washed with brine, dried over
anhydrous sodium sulfate,
filtered and concentrated under reduced pressure. The residue was purified by
column
chromatography (silica gel, eluent: petroleum ether! ethyl acetate 10:1).
Yield: 5.7 g (55% of theory).
1H-NMR (300 MHz, DMSO-d6): 6 [ppm] = 7.78 (d, 1H), 7.47 (dd, 1H), 7.37 (d,
1H), 6.44 (br s, 1H),
5.12 (t, 1H), 4.05-3.98 (m, 2H), 1.67 (s, 3H).
Example 2.1C
3 -(2-Bromo -4 -chloropheny1)-2 -methylpropan-1 -ol (racemate)
CI Br
CH 3
OH
Tris(triphenylphosphine)rhodium(I) chloride (3.2 g, 3.4 mmol, 0.3 eq.) was
added at RT under
nitrogen atmosphere to a mixture of 3-(2-bromo-4-chloropheny1)-2-methylprop-2-
en-1-ol (E/Z
mixture) (3.0 g, 11.5 mmol, 1.0 eq.) in ethanol (30 m1). The resulting mixture
was purged with
hydrogen gas and stirred at RT for 48 h under hydrogen gas atmosphere (2 bar).
The reaction mixture
was filtered through Celite and the filtrate concentrated under reduced
pressure. The residue was
purified by column chromatography (silica gel, eluent: petroleum ether! ethyl
acetate 20:1). Yield:
1.90 g (59% of theory).
1H-NMR (300 MHz, DMSO-d6): 6 [ppm] = 7.71 (d, 1H), 7.41 (dd, 1H), 7.33 (d,
1H), 4.57 (t, 1H),
3.32-3.26 (m, 2H, partially concealed), 2.85-2.76 (m, 1H), 2.49-2.39 (m, 1H),
1.93-1.79 (m, 1H),
0.82 (d, 3H).
Example 2.2A
(2-Bromo -4 -chloro-3 -fluorophenyl)methanol
Br HO
CI 1101
2-Bromo-4-chloro-3-fluorobenzoic acid (5.00 g, 19.7 mmol, 1.0 eq.) was
dissolved in
tetrahydrofuran (150 ml), cooled to 0 C and a solution of borane
tetrahydrofuran complex (59 ml,
1.0 M in tetrahydrofuran, 59 mmol, 3.0 eq.) was added dropwise. The mixture
was stirred at 0 C for
1 h and at RT for 2 days. Further amounts of borane tetrahydrofuran complex
solution (30 ml, 1.0 M
in tetrahydrofuran, 30 mmol, 1.5 eq.) were added at 0 C and stirring was
continued at RT overnight.
The reaction mixture was diluted with ethyl acetate and saturated aqueous
sodium bicarbonate
solution, extracted with ethyl acetate, washed with brine, dried over
anhydrous sodium sulfate and
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filtered. The filtrate was concentrated under reduced pressure and the crude
product was used in the
subsequent step without further purification. Yield: 4.49 g (95% of theory).
1H-NMR (400 MHz, DMSO-d6): 6 [ppm] = 7.71-7.58 (m, 1H), 7.44-7.32 (m, 1H),
5.67-5.56 (t, 1H),
4.58-4.44 (d, 2H).
Example 2.2B
2-Bromo-1-(bromomethyl)-4-chloro-3 -fluorobenzene
Br Br
CI 1.1
To a solution of (2-bromo-4-chloro-3-fluorophenyl)methanol (9.16 g, 66%
purity, 25.2 mmol, 1.0
eq.) in diethyl ether (120 ml) at 0-5 C was added a solution of phosphorous
tribromide (3.6 ml, 38
mmol, 1.5 eq.) in diethyl ether (40 m1). The mixture was stirred at RT for 4
days, followed by the
addition of further amounts of phosphorous tribromide (12 ml, 130 mmol, 5.0
eq.) at 0 C. Stirring
was continued at RT for 0.5 h, followed by the addition of saturated aqueous
sodium bicarbonate
solution at 0 C. The mixture was extracted with dichloromethane and ethyl
acetate. The combined
organic layers were washed with brine, dried over anhydrous sodium sulfate and
filtered. The filtrate
was concentrated under reduced pressure and the crude product was used in the
subsequent step
without further purification. Yield: 10.26 g (quantitative of theory).
1H-NMR (400 MHz, DMSO-d6): 6 [ppm] = 7.74-7.59 (m, 1H), 7.57-7.47 (m, 1H),
4.81-4.73 (s, 2H).
Example 2.2C
Diethyl R2-bromo-4-chloro-3-fluorophenyl)methyl] (methyl)propanedioate
Br
C H
C 113o_ =-=
CI 0 0
(C H 3
2-Bromo-1-(bromomethyl)-4-chloro-3-fluorobenzene (10.3 g, 33.9 mmol, 1.0 eq.),
diethyl
methylmalonate (6.21 g, 35.6 mmol, 1.1 eq.) and caesium carbonate (13.3 g,
40.7 mmol, 1.2 eq.)
were dissolved in N,N-dimethylformamide (30 ml) and stirred at 70 C overnight.
The mixture was
diluted with water, extracted with ethyl acetate, washed with brine, dried
over anhydrous sodium
sulfate and filtered. The filtrate was concentrated under reduced pressure and
the crude product was
used in the subsequent step without further purification. Yield: 11.81 g (54%
purity, 48% of theory).
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GC-MS (method 6): Rt = 7.41 min; MS (APCIpos): m/z = 397 [M-411+
Example 2.2D
R2-Bromo-4-chloro-3-fluorophenyl)methyll(methyl)propanedioic acid
Br
C H30 H
CI 0 0
0 H
Diethyl R2-bromo-4-chloro-3-fluorophenyl)methyll(methyl)propanedioate (11.8 g,
54% purity,
16.1 mmol, 1.0 eq.) was dissolved in ethanol (100 ml), an aqueous solution of
sodium hydroxide
(97 ml, 1.0 M, 97 mmol, 6.0 eq.) was added and the mixture was stirred at RT
overnight and at 80 C
for 3 days. The mixture was concentrated under reduced pressure, the residue
was treated with
aqueous hydrogen chloride (1 N) and extracted with dichloromethane and ethyl
acetate. The
combined organic phases were dried over anhydrous sodium sulfate, filtered and
concentrated under
reduced pressure. The crude product was used in the subsequent step without
further purification.
Yield: 7.01 g (50% purity, 64% of theory).
LC-MS (method 1): Rt = 0.85 min; MS (ESIneg): m/z = 339 EM-Ht
Example 2.2E
3-(2-Bromo-4-chloro-3-fluoropheny1)-2-methylpropanoic acid (racemate)
Br 0
0 H
C H 3
C
R2-Bromo-4-chloro-3-fluorophenyl)methyll(methyl)propanedioic acid (7.00 g, 50%
purity,
10.3 mmol, 1.0 eq.) was dissolved in acetic acid (100 ml) and stirred at 120 C
overnight. The mixture
was then concentrated under reduced pressure, dissolved in acetonitrile and
water, filtered and the
filtrate was purified by preparative HPLC (reversed phase, eluent: water with
0.05% formic acid /
acetonitrile 90:10 to 5:95). Yield: 2.52 g (77% purity, 64% of theory).
LC-MS (method 1): R1= 1.03 min; MS (ESIneg): m/z = 293 EM-Ht
Example 2.2F
3 -(2-Bromo -4 -chloro -3 -fluoropheny1)-2-methylpropan-1-ol (racemate)
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Br
OH
CH 3
C I
3-(2-Bromo-4-chloro-3-fluoropheny1)-2-methylpropanoic acid (racemate) (2.40 g,
77% purity,
6.25 mmol, 1.0 eq.) was dissolved in tetrahydrofuran (60 ml), cooled to 0 C
and a solution of borane
tetrahydrofuran complex (19 ml, 1.0 M in tetrahydrofuran, 19 mmol, 3.0 eq.)
was added dropwise.
The mixture was stirred at 0 C for 1 h and at RT overnight. The reaction
mixture was diluted with
ethyl acetate and saturated aqueous sodium bicarbonate solution, extracted
with ethyl acetate, washed
with brine, dried over anhydrous magnesium sulfate and filtered. The filtrate
was concentrated under
reduced pressure and the crude product was purified by column chromatography
(silica gel, eluent:
cyclohexane / ethyl acetate 99:1 to 40:60). Yield: 1.65 g (87% purity, 82% of
theory).
1H-NMR (500 MHz, DMSO-d6): 6 [ppm] = 7.59-7.47 (m, 1H), 7.24-7.11 (m, 1H),
4.58 (t, 1H), 3.31-
3.21 (m, 2H), 2.90-2.82 (m, 1H), 2.49-2.46 (m, 1H), 1.96-1.79 (m, 1H), 0.82
(d, 3H).
Example 2.3A
2-[(2-Bromo-4-chlorophenyl)methylidenelbutanal (E/Z mixture)
CI Br CH3
Lcro
To a solution of 2-bromo-4-chlorobenzaldehyde (10.0 g, 45.6 mmol, 1.0 eq.) and
sodium hydroxide
(182 mg, 4.56 mmol, 0.1 eq.) in methanol (40 ml) was added butanal (4.5 ml, 50
mmol, 1.1 eq.)
dropwise at RT. The mixture was stirred at RT overnight, followed by addition
of water. The mixture
was partially concentrated under reduced pressure, dried over anhydrous sodium
sulfate, filtered and
washed with ethyl acetate. The filtrate was concentrated under reduced
pressure. The crude mixture
was used in the subsequent step without further purification. Yield: 12.8 g
(86% purity, 78% of
theory).
1H-NMR (500 MHz, DMSO-d6): 6 [ppm] = 9.67 (s, 1H), 7.93 (d, 1H), 7.59 (dd,
1H), 7.54-7.42 (m,
2H), 2.26 (q, 2H), 0.96 (t, 3H).
Example 2.3B
2-[(2-Bromo-4-chlorophenyl)methylidenelbutan-1-ol (E/Z mixture)
CI Br CH3
(101 OH
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To a suspension of sodium borohydride (2.68 g, 70.9 mmol, 2.0 eq.) in
tetrahydrofuran (10 ml) and
methanol (10 ml) was added a solution of 2-[(2-bromo-4-
chlorophenyl)methylidenelbutanal (E/Z
mixture) (11.3 g, 86% purity, 35.5 mmol, 1.0 eq.) in tetrahydrofuran (90 ml)
dropwise at 0 C. The
resulting mixture was stirred at RT for 3 h and cooled to 0 C, followed by the
addition of aqueous
hydrogen chloride (1 N) to adjust the pH value to 6. The aqueous phase was
extracted with ethyl
acetate and the combined organic layers were washed with brine, dried over
anhydrous sodium
sulfate and filtered. The filtrate was concentrated under reduced pressure and
the crude product was
used in the subsequent step without further purification. Yield: 10.58 g (86%
purity, 93% of theory).
1H-NMR (500 MHz, DMSO-d6): 6 [ppm] = 7.73 (d, 1H), 7.43 (dd, 1H), 7.28 (d,
1H), 6.38 (s, 1H),
4.09-3.94 (m, 2H), 2.06 (q, 2H), 1.98 (s, 1H), 0.96 (t, 3H).
Example 2.3C
2-(2-Bromo-4-chlorobenzyl)butan-1-ol (racemate)
CI Br C H 3
OH
To a solution of 2-[(2-bromo-4-chlorophenyl)methylidenelbutan-1-ol (E/Z
mixture) (10.6 g, 86%
purity, 33.0 mmol, 1.0 eq.) in dichloromethane (200 ml) was added
tris(triphenylphosphine)rhodium(I) chloride (1.53 g, 1.65 mmol, 0.1 eq.) under
argon atmosphere.
The resulting mixture was purged with hydrogen gas and stirred at 22 C for 48
h under a hydrogen
atmosphere at 50 bar. The reaction mixture was filtered through Celite and
concentrated under
reduced pressure. The residue was purified by column chromatography (silica
gel, eluent:
cyclohexane / ethyl acetate 100:0 to 65:35). Yield: 8.84 g (77% purity, 74% of
theory).
1H-NMR (600 MHz, DMSO-d6): 6 [ppm] = 7.68 (d, 1H), 7.39 (dd, 1H), 7.35-7.28
(m, 1H), 4.46 (t,
1H), 3.32-3.24 (m, 2H), 2.70-2.65 (m, 1H), 2.63-2.57 (m, 1H), 1.69-1.56 (m,
1H), 1.44-1.35 (m, 1H),
1.24 (s, 1H), 0.86 (t, 3H).
Example 2.4A
3-(2-Bromo-4-chloropheny1)-2-methylpropanal (racemate)
Br
0
C H 3
CI
To a solution of 3-(2-bromo-4-chloropheny1)-2-methylpropan-1-ol (racemate)
(5.0 g, 18.4 mmol,
1.0 eq.) in dichloromethane (100 ml) were added 3,3,3-triacetoxy-3-
iodophthalide (19.5 g, 46.0
mmol, 2.5 eq.) and sodium hydrogen carbonate (11.6 g, 138.0 mmol, 7.5 eq.) at
0 C. After stirring
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at RT for 3 h, the reaction mixture was quenched with a mixture (1:1) of a
saturated aqueous solution
of sodium thiosulfate and a saturated aqueous solution of sodium carbonate and
extracted with
dichloromethane. The combined organic layers were washed with water and brine,
dried over
anhydrous magnesium sulfate, filtered and concentrated under reduced pressure.
The residue was
purified by column chromatography (silica gel, eluent: petroleum ether! ethyl
acetate 97:3). Yield:
3.90 g (85% purity, 68% of theory).
1H-NMR (400 MHz, DMSO-d6): 6 [ppm] = 9.66 (s, 1H), 7.74 (d, 1H), 7.44-7.37 (m,
2H), 3.16-3.11
(m, 1H), 2.78-2.64 (m, 2H), 1.01 (d, 3H).
Example 2.4B
4-(2-Bromo-4-chloropheny1)-3-methylbutan-2-ol (mixture of stereoisomers)
Br OH
C H 3
HC
CI
To a solution of 3-(2-bromo-4-chloropheny1)-2-methylpropanal (racemate) (4.2
g, 90% purity,
14.4 mmol, 1.0 eq.) in tetrahydrofuran (80 ml) was added methylmagnesium
bromide (14.5 ml, 3.0
M in 2-methyl tetrahydrofuran, 43.3 mmol, 3.0 eq.) at 0 C under nitrogen
atmosphere. After stirring
at 0 C for 2 h, the reaction mixture was quenched with water and stirred for
10 min. The mixture
was then extracted with ethyl acetate, washed with water and brine, dried over
anhydrous sodium
sulfate, filtered and concentrated under reduced pressure. The residue was
purified by column
chromatography (silica gel, eluent: petroleum ether! ethyl acetate 87:13) to
give 2.8 g of 4-(2-bromo-
4-chloropheny1)-3-methylbutan-2-ol (mixture of stereoisomers) in 63% purity.
In order to remove
side products, the crude product was dissolved in dichloromethane (15 ml),
treated with imidazole
(0.66 g, 9.7 mmol, 0.7 eq.) and tert-butyldimethylsilyl chloride (0.73 g, 4.8
mmol, 0.35 eq.), stirred
at RT for 2 h and filtered through Celite . The filtrate was concentrated
under reduced pressure. The
residue was purified by column chromatography (silica gel, eluent: n-hexane /
ethyl acetate 94:6).
Yield: 1.47 g (83% purity, 30% of theory).
1H-NMR (400 MHz, DMSO-d6): 6 [ppm] = 7.71-7.68 (m, 1H), 7.41-7.37 (m, 1H),
7.35-7.28 (m,
1H), 4.51-4.45 (m, 1H), 3.57-3.50 (m, 1H), 2.93-2.79 (m, 1H), 2.49-2.34 (m,
1H), 1.77-1.68 (m, 1H),
1.16-1.05 (m, 3H), 0.75-0.67 (m, 3H).
Example 2.5A
2-Cyclobutylidene -1,1 -dimethylhydrazine
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µ9
N C H
1µ1' 3
C H 3
A solution of cyclobutanone (1.00 g, 14.3 mmol) and 1,1-dimethylhydrazine (1.1
ml, 14 mmol, 1.0
eq.) in tetrahydrofuran (6.0 ml) was stirred at 60 C for 2.5 h under microwave
irradiation. All
volatiles were removed at 30 C in vacuo (>50 mbar). The residue was purified
by Kugelrohr
distillation (75 C, 20 mbar). Yield: 1.08 g (67% of theory).
GC-MS (method 2): R1= 1.79 min; m/z = 112 [M]
1H-NMR (400 MHz, CDC13): 6 [ppm] = 3.02-2.87 (m, 4H), 2.58 (s, 6H), 1.98
(quin, 2H).
Example 2.5B
2-[(2 -B romo -4-chlorophenyl)methyl] cyclobutan-1 -one (racemate)
Br
0
C
I
A solution of n-butyllithium (6.1 ml, 1.6 M in hexane, 9.8 mmol, 1.1 eq.) was
added under argon
atmosphere at -10 C to a solution of 2-cyclobutylidene-1,1-dimethylhydrazine
(1.00 g, 8.92 mmol)
in tetrahydrofuran (40 m1). The reaction mixture was stirred at -10 C for 1 h,
followed by the addition
of a solution of 2-bromo-1-(bromomethyl)-4-chlorobenzene (3.13 g, 81% purity,
8.92 mmol, 1.0 eq.)
in tetrahydrofuran (10 m1). The reaction mixture was stirred at RT for 20 h,
diluted with aqueous
hydrochloric acid (2 N) and stirred at RT for 1 h. After addition of ethyl
acetate and phase separation,
the aqueous phase was extracted with ethyl acetate. The combined organic
phases were washed with
brine, dried over anhydrous sodium sulfate, filtered and concentrated under
reduced pressure. The
crude product was purified by column chromatography (silica gel, eluent:
cyclohexane / ethyl acetate
mixtures). Yield: 1.89 g (77% of theory).
1H-NMR (400 MHz, DMSO-d6): 6 [ppm] = 7.72 (d, 1H), 7.43 (dd, 1H), 7.38 (d,
1H), 3.73-3.61 (m,
1H), 3.15-2.97 (m, 2H), 2.95-2.81 (m, 2H), 2.06 (qd, 1H), 1.77-1.64 (m, 1H).
Example 2.5C
trans-2- [(2-Bromo -4 -chlorophenyl)methyll cyclobutan-l-ol (racemate)
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Br
OH
CI
Sodium borohydride (505 mg, 13.3 mmol, 1.0 eq.) was added at 0 C to a solution
of 2-[(2-bromo-4-
chlorophenyl)methyllcyclobutan-1-one (racemate) (3.65 g, 13.3 mmol) in a
mixture of
dichloromethane / methanol (2:1, 67 m1). The reaction mixture was stirred at 5
C for 1 h and diluted
with water. After phase separation, the aqueous phase was extracted with
dichloromethane. The
combined organic phases were dried over anhydrous sodium sulfate, filtered and
concentrated under
reduced pressure. The crude product was purified by column chromatography
(silica gel, eluent:
cyclohexane / ethyl acetate mixtures). Yield: 1.03 g (27% of theory) of the
cis-product and 2.67 g
(72% of theory) of the desired trans-product (the title compound Example
2.5C).
LC-MS (method 4): Rt = 2.00 min; MS (ESIpos): m/z = 299 [M+Nal+
1H-NMR (400 MHz, DM50-d6): 6 [ppm] = 7.68 (d, 1H), 7.39 (dd, 1H), 7.36 (d,
1H), 5.03 (d, 1H),
3.71 (quin, 1H), 2.91 (dd, 1H), 2.69 (dd, 1H), 2.35-2.20 (m, 1H), 2.08-1.99
(m, 1H), 1.65-1.51 (m,
2H), 1.15-1.01 (m, 1H).
Example 2.6A
Ethyl 3-(2-bromo-4-chlorophenyl)prop-2-enoate (E/Z mixture)
CI
OC H 3
Br 0
While cooling in an iced water bath,
ethoxycarbonylmethylenetriphenylphosphorane (47.6 g,
136.7 mmol, 1.5 eq.) was added dropwise to a solution of 2-bromo-4-
chlorobenzaldehyde (20.0 g,
91.1 mmol) in dichloromethane (200 m1). After stirring at RT for 2 h, the
reaction mixture was
concentrated under reduced pressure. The residue was purified by column
chromatography (silica
gel, eluent: petroleum ether! ethyl acetate 6:1). Yield: 26.0 g (98% of
theory).
1H-NMR (400 MHz, DMSO-d6): 6 [ppm] = 7.94 (d, 0.8H), 7.84 (d, 0.8 H), 7.80 (d,
1H), 7.50 (dd,
0.8H), 7.45 (dd, 0.2H), 7.42 (d, 0.2H), 7.06 (d, 0.2H), 6.68 (d, 0.8H), 6.18
(d, 0.2H), 4.21 (q, 1.6H),
4.04 (q, 0.4H), 1.26 (t, 2.4H), 1.10 (t, 0.6H).
.. Example 2.6B
Ethyl 3-(2-bromo-4-chlorophenyl)propanoate
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CI
0 H 3
Br 0
Chlorotris(triphenylphosphine)rhodium(I) (12.3 g, 13.3 mmol, 0.15 eq.) was
added under nitrogen
atmosphere to a solution of ethyl 3-(2-bromo-4-chlorophenyl)prop-2-enoate (E/Z
mixture) (26.0 g,
88.9 mmol) in ethanol (250 m1). The resulting mixture was purged with hydrogen
for 5 min and
stirred at RT for 12 h under hydrogen atmosphere (2 bar). The mixture was
filtered through Celite
and the filtrate concentrated under reduced pressure. The residue was re-
dissolved in ethanol
(100 ml), followed by the addition of chlorotris(triphenylphosphine)rhodium(I)
(6.2 g, 6.7 mmol,
0.075 eq.) under nitrogen atmosphere. The resulting mixture was purged with
hydrogen for 5 min
and stirred at RT for 15 h under hydrogen atmosphere (2 bar). The reaction
mixture was filtered
through Celite and the filtrate concentrated under reduced pressure. The
residue was purified by
column chromatography (silica gel, eluent: 100% petroleum ether). Yield: 25.6
g (97% of theory).
1H-NMR (400 MHz, DMSO-d6): 6 [ppm] = 7.73 (d, 1H), 7.43 (dd, 1H), 7.38 (d,
1H), 4.06 (m, 2H),
2.95 (t, 2H), 2.62 (t, 2H), 1.17 (t, 3H).
Example 2.6C
3-(2-Bromo-4-chlorophenyl)propanoic acid
CI
OH
Br 0
A solution of sodium hydroxide (31.9 g, 798.4 mmol, 4.0 eq.) in water (250 ml)
was added to a stirred
solution of ethyl 3-(2-bromo-4-chlorophenyl) propanoate (60.0 g, 199.6 mmol)
in ethanol (400 m1).
After stirring at RT for 15 h, the reaction mixture was concentrated under
reduced pressure. The
mixture was adjusted to pH 3 with aqueous hydrochloric acid (4 N) and
extracted with ethyl acetate.
The combined organic layers were dried over anhydrous sodium sulfate, filtered
and evaporated to
dryness. Yield: 54.0 g (98% of theory).
1H-NMR (300 MHz, DMSO-d6): 6 [ppm] = 12.25 (s, 1H), 7.71 (s, 1H), 7.45-7.34
(m, 2H), 2.91 (t,
2H), 2.53 (t, 2H).
Example 2.6D
(4R)-3 43 -(2 -B romo -4-chlorophenyl)propanoyll -4-pheny1-1,3 -oxazolidin-2 -
one (single
stereoisomer)
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CI 0
0
Br 0 *
NN-Dimethylformamide (0.62 g, 8.5 mmol, 0.1 eq.) was added to a suspension of
3-(2-bromo-4-
chlorophenyl)propanoic acid (22.5 g, 85.4 mmol) in thionyl chloride (60 m1).
After stirring at 80 C
for 4 h, the reaction mixture was evaporated to dryness to give 24.0 g of 3-(2-
bromo-4-chlorophenyl)
propanoyl chloride.
A solution of n-butyllithium (35.7 ml, 2.5 M in hexane, 89.4 mmol, 1.05 eq.)
was added at -78 C
under nitrogen atmosphere to a stirred solution of (4R)-4-phenyl-1,3-
oxazolidin-2-one (single
stereoisomer) (13.9 g, 85.1 mmol, 1.0 eq.) in tetrahydrofuran (300 m1). The
resulting mixture was
stirred at -78 C for 30 min, followed by the addition of a solution of 3-(2-
bromo-4-
chlorophenyl)propanoyl chloride (24.0 g, 85.1 mmol) in tetrahydrofuran (200
ml) at -78 C. After
stirring at RT for 5 h, the reaction mixture was quenched with saturated
aqueous solution of sodium
bicarbonate and extracted with ethyl acetate. The combined organic layers were
washed with brine,
dried over anhydrous sodium sulfate, filtered and concentrated under reduced
pressure. The residue
was purified by column chromatography (silica gel, eluent: petroleum ether /
ethyl acetate 10:1).
Yield: 29.5 g (85% of theory).
1H-NMR (400 MHz, DMSO-d6): 6 [ppm] = 7.71 (d, 1H), 7.44-7.25 (m, 7H), 5.46
(dd, 1H), 4.74 (t,
1H), 4.17 (dd, 1H), 3.28-3.11 (m, 2H), 2.99-2.85 (m, 2H).
Example 2.6E
(4R)-3-1(2R)-24(2-Bromo-4-chlorophenyl)methyll -3,3,3 -trifluoropropanoyl} -4 -
phenyl-1,3 -
oxazolidin-2-one (single stereoisomer)
CI FO
FO
Br 0 *
A solution of lithium bis(trimethylsilyl)amide (75.8 ml, 1.0 M in
tetrahydrofuran, 75.8 mmol,
1.05 eq.) was added at -78 C under nitrogen atmosphere to a stirred solution
of (4R)-343-(2-bromo-
4-chlorophenyl)propanoy11-4-pheny1-1,3-oxazolidin-2-one (single
stereoisomer) (29.5 g,
72.2 mmol) in tetrahydrofuran (300 m1). After stirring at -78 C for 30 min, 1-
trifluoromethy1-1,2-
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benziodoxo1-3(1H)-one (24.0 g, 75.8 mmol, 1.05 eq.) was added to the mixture
at -78 C. The
resulting mixture was allowed to warm to RT during 1 h and stirred further 4
h. The reaction mixture
was quenched with saturated aqueous solution of sodium carbonate and extracted
with ethyl acetate.
The combined organic layers were washed with water and brine, dried over
anhydrous sodium
sulfate, filtered and concentrated under reduced pressure to give 22.5 g of
crude product, which was
triturated with methanol and stirred for 30 min. The solid was collected by
filtration, washed with
methanol and dried under air atmosphere. Yield: 16.0 g (46% of theory).
LC-MS (method 10): Rt = 1.26 min; MS (ESIpos): m/z = 478 [M+I-11+
1H-NMR (400 MHz, DMSO-d6): 6 [ppm] = 7.80 (d, 1H), 7.48 (dd, 1H), 7.44-7.37
(m, 2H), 7.37-7.31
(m, 2H), 7.30-7.24 (m, 2H), 5.50-5.37 (m, 2H), 4.66 (t, 1H), 4.20 (dd, 1H),
3.34-3.16 (m, 2H).
19F-NMR (376 MHz, DMSO-d6): 6 [ppm] = -65.71 (s, 3F).
Example 2.6F
(2R)-2-[(2-Bromo-4-chlorophenyl)methyll-3,3,3-trifluoropropan-1-ol (single
stereoisomer)
CI F F
411 OH
Br
Sodium borohydride (3.4 g, 90.4 mmol, 5.0 eq.) was added at 0 C to a stirred
solution of (4R)-3-
[(2R)-2 -(2-bromo -4-chlorobenzy1)-3 ,3 ,3 -trifluoropropanoyl] -4 -phenyl -
1,3 -oxazol idin-2 -one (single
stereoisomer) (8.9 g, 18.1 mmol) in tetrahydrofuran (60 ml) and water (20 m1).
After stirring at RT
for 5 h, the reaction mixture was quenched with saturated aqueous solution of
ammonium chloride
and extracted with ethyl acetate. The combined organic layers were washed with
water and brine,
dried over anhydrous sodium sulfate and filtered. The filtrate was
concentrated under reduced
pressure. The residue was purified by column chromatography (silica gel,
eluent: petroleum ether /
ethyl acetate 5:1). Yield: 4.5 g (75% of theory).
Chiral SFC: Rt = 1.11 min, >99% ee
Analysis method: SFC: column: Chiralpak AD-3 3 um, 100 mm x 3 mm, eluent:
carbon dioxide (A),
methanol with 0.1% DEA (B), gradient: 5% to 20% B in 2 min, hold 1 min at 20%
B; temperature:
C; flow rate: 2.0 ml/min; UV detection: 254 nm.
1H-NMR (400 MHz, DMSO-d6): 6 [ppm] = 7.75 (d, 1H), 7.50-7.40 (m, 2H), 5.02 (t,
1H), 3.64-3.44
(m, 2H), 3.03-2.93 (m, 2H), 2.75-2.59 (m, 1H).
19F-NMR (376 MHz, DMSO-d6): 6 [ppm] = -67.19 (s, 3F).
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Example 2.7A
tert-Butyl (4-chloro-3-fluorophenyl)carbamate
CI
401 3)<C H 3
N 0 CH 3
To a mixture of 4-chloro-3-fluoroaniline (15.0 g, 103 mmol, 1.0 eq.) in water
(150 ml) was added
di-tert-butyl dicarbonate (26 ml, 110 mmol, 1.1 eq.). The resulting mixture
was stirred at RT for
24 h, extracted with diethyl ether, dried over anhydrous sodium sulfate,
filtered and concentrated
under reduced pressure. The crude product was used in the subsequent reaction
without further
purification. Yield: 21.6 g (85% of theory).
LC-MS (method 1): R1= 1.12 min; MS (ESIneg): m/z = 244 EM-Hr
Example 2.7B
tert-Butyl (2-bromo-4-chloro-3-fluorophenyl)carbamate
CI
0 H3C
)<CH3
N 0 CH 3
Br
A solution of tert-butyllithium (88 ml, 1.7 M in pentane, 150 mmol, 1.5 eq.)
in tetrahydrofuran (350
ml) was cooled to -78 C, a solution of tert-butyl (4-chloro-3-
fluorophenyl)carbamate (24.4 g, 99.4
mmol, 1.0 eq.) in tetrahydrofuran (150 ml) was added dropwise over a period of
90 min. Stirring was
continued at this temperature for 60 min, before 1,2-dibromoethane (10 ml, 120
mmol, 1.2 eq.) was
added dropwise. The resulting mixture was stirred at -78 C for 1.5 h, at RT
overnight and then treated
with a saturated aqueous solution of ammonium chloride, extracted with ethyl
acetate, dried over
anhydrous sodium sulfate, filtered and concentrated under reduced pressure.
The crude mixture was
purified by column chromatography (silica gel, eluent: cyclohexane / ethyl
acetate 20:1 to 10:1).
Yield: 15.9 g (49% of theory).
LC-MS (method 3): Rt = 4.20 min; MS (ESIneg): m/z = 323 EM-Hr
Example 2.7C
2-Bromo-4-chloro-3-fluoroaniline trifluoroacetate
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CI
1.1 N H 2 X TFA
Br
To a solution of tert-butyl (2-bromo-4-chloro-3-fluorophenyl)carbamate (15.9
g, 49.0 mmol, 1.0 eq.)
in dichloromethane (200 ml) was added trifluoroacetic acid (19 ml, 240 mmol,
5.0 eq.) and the
resulting mixture was stirred at RT overnight. The mixture was then
concentrated under reduced
pressure and the remaining residual was dissolved for two times in
dichloromethane followed by
evaporation. The crude product was used in the subsequent reaction without
further purification.
Yield: 15.1 g (87% of theory).
LC-MS (method 15): Rt = 1.62 min; MS (ESIneg): m/z = 222 [M-H-TFAI
Example 2.7D
2-Bromo-4-chloro-3-fluoro-1-iodobenzene
CI
F 1.1 I
Br
A solution of 2-bromo-4-chloro-3-fluoroaniline trifluoroacetate in water (320
ml) was cooled to 0 C,
followed by the dropwise addition of sulfuric acid (60 m1). A solution of
sodium nitrite (3.27 g,
47.4 mmol, 1.1 eq.) in water (30 ml) was added to the mixture and the
resulting suspension was
stirred for 1 h while the temperature was kept at 5-10 C. Subsequently, a
solution of potassium iodide
(9.38 g, 56.5 mmol, 1.3 eq.) in water (50 ml) was added under vigorous
stirring. The reaction mixture
was allowed to warm to RT and extracted with ethyl acetate. The combined
organic phases were
washed with aqueous solutions of sodium hydroxide (1 N), sodium thiosulfate (1
N), hydrogen
chloride (1 N) and a saturated aqueous sodium bicarbonate solution. The washed
organic phases were
dried over anhydrous sodium sulfate, filtered and concentrated under reduced
pressure. The crude
mixture was purified by column chromatography (silica gel, eluent: cyclohexane
/ ethyl acetate 50:1).
Yield: 12.1 g (82% of theory).
1H-NMR (400 MHz, DM50-c/6): 6 [ppm] = 7.81 (dd, 1H), 7.43 (dd, 1H).
Example 2.7E
3 -(2-Bromo -4 -chloro -3 -fluorophenyl)propanal
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CI
0
Br
2-Bromo-4-chloro-3-fluoro-1-iodobenzene (9.00 g, 26.3 mmol, 1.0 eq.),
palladium(II) acetate
(118 mg, 526 umol, 0.02 eq.), tetra-N-butylammonium chloride (7.31 g, 26.3
mmol, 1.0 eq.) and
sodium bicarbonate (5.52 g, 65.8 mmol, 2.5 eq.) were dissolved in N,N-
dimethylformamide (60 ml)
.. and argon was passed through the resulting suspension for 10 min. After
addition of prop-2-en-1-ol
(2.7 ml, 39 mmol, 1.5 eq.), the mixture was heated to 40 C and stirred at this
temperature overnight,
subsequently followed by the addition diethyl ether and water. The aqueous
phase was extracted with
diethyl ether and the combined organic phases were washed with water and
brine, dried over
anhydrous sodium sulfate, filtered and concentrated under reduced pressure.
The crude mixture was
.. used in the subsequent step without further purification. Yield: 6.29 g
(82% purity, 74% of theory).
1H-NMR (400 MHz, DMSO-d6): 6 [ppm] = 9.72 (s, 1H), 7.56 (dd, 1H), 7.25 (dd,
1H), 2.98 (t, 2H),
2.81 (t, 2H).
Example 2.7F
(2R)-2-(2-Bromo-4-chloro-3-fluorobenzy1)-3,3,3-trifluoropropan-1-ol (single
stereoisomer)
CI F F
0 H
Br
Argon was passed through a mixture of 3,3-dimethy1-1-(trifluoromethyl)-1,2-
benziodoxole (3.53 g,
10.7 mmol, 1.0 eq.), (5R)-5-benzy1-2,2,3-trimethy1-4-oxoimidazolidin-1-ium
trifluoroacetate (single
stereoisomer) (711 mg, 2.14 mmol, 0.2 eq.) and copper(I) chloride (52.9 mg,
535 umol, 0.05 eq.) for
10 min. The mixture was cooled to -20 C before a solution of 3-(2-bromo-4-
chloro-3-
.. fluorophenyl)propanal (4.50 g, 82% purity, 13.9 mmol, 1.3 eq.) in
chloroform (30 ml) was added.
The resulting mixture was stirred at -20 C for 5 h and kept in the freezer at -
18 C overnight.
Additional amounts of 3,3-dimethy1-1-(trifluoromethyl)-1,2-benziodoxole (2.12
g, 6.41 mmol, 0.6
eq.), (5R)-5-benzy1-2,2,3-trimethy1-4-oxoimidazolidin-1-ium trifluoroacetate
(single stereoisomer)
(355 mg, 1.07 mmol, 0.1 eq.) and copper(I) chloride (265 mg, 2.67 mmol, 0.25
eq.) were added and
.. the mixture was stirred at -20 C for additional 5.5 h. After cooling to -78
C, precooled
dichloromethane (45 ml, -78 C) was added and the mixture was treated with
sodium borohydride
(4.04 g, 107 mmol, 10.0 eq.) and precooled methanol (21 ml, -78 C). Stirring
was continued at -78 C
for 1 h, followed by the addition of a saturated aqueous solution of ammonium
chloride and warming
of the reaction mixture to RT. The aqueous phase was then extracted with
dichloromethane and the
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combined organic phases were dried over anhydrous sodium sulfate, filtered and
concentrated under
reduced pressure. The crude mixture was purified by preparative HPLC (reversed
phase, eluent:
water with 0.05% formic acid / acetonitrile 90:10 to 5:95). Yield: 3.68 g (76%
of theory).
1H-NMR (400 MHz, DMSO-d6): 6 [ppm] = 7.61 (dd, 1H), 7.33 (dd, 1H), 5.03 (t,
1H), 3.62-3.46 (m,
2H), 3.10-2.99 (m, 2H), 2.76-2.64 (m, 1H).
Example 2.8A
3 -(4-Chloro -3 -fluorophenyl)propanal
CI
F 401
0
1-Chloro-2-fluoro-4-iodobenzene (10.0 g, 39.0 mmol, 1.0 eq.), tetra-N-
butylammonium chloride
(10.8 g, 39.0 mmol, 1.0 eq.) and sodium bicarbonate (8.19 g, 97.5 mmol, 2.5
eq.) were dissolved in
N,N-dimethylformamide (80 ml) and argon was passed through the resulting
suspension for 10 min.
After addition of palladium(II) acetate (175 mg, 780 lama 0.02 eq.) and prop-2-
en-1-ol (4.0 ml, 58
mmol, 1.5 eq.), the mixture was heated to 50 C and stirred at this temperature
for 4 h, followed by
the addition of methyl tert-butyl ether and water. After phase separation, the
aqueous phase was
extracted with methyl tert-butyl ether. The combined organic phases were
washed with water and
brine, dried over anhydrous sodium sulfate, filtered and concentrated under
reduced pressure. The
crude mixture was used in the subsequent step without further purification.
Yield: 6.55 g (90% of
theory).
1H-NMR (600 MHz, DMSO-d6): 6 [ppm] = 9.70 (t, 1H), 7.51-7.40 (m, 1H), 7.31-
7.27 (m, 1H), 7.10
(dd, 1H), 2.89-2.84 (m, 2H), 2.83-2.77 (m, 2H).
Example 2.8B
(2R)-2-(4-Chloro-3-fluorobenzy1)-3,3,3-trifluoropropanal (single stereoisomer)
CI F F
0
Argon was passed through a mixture of 3,3-dimethy1-1-(trifluoromethyl)-1,2-
benziodoxole (10.6 g,
32.2 mmol, 1.0 eq.), (5R)-5-benzy1-2,2,3-trimethy1-4-oxoimidazolidin-1-ium
trifluoroacetate (single
stereoisomer) (2.14 g, 6.43 mmol, 0.2 eq.) and copper(I) chloride (239 mg,
2.41 mmol, 0.075 eq.)
for 10 min. The mixture was cooled to -20 C before a solution of 3-(4-chloro-3-
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fluorophenyl)propanal (6.00 g, 32.2 mmol, 1.0 eq.) in chloroform (70 ml) was
added. The resulting
mixture was kept in the freezer at -18 C for 72 h and stirred at -20 C for
additional 4 h. The mixture
was then immediately purified by column chromatography (silica gel, eluent:
dichloromethane /
methanol 98:2 to 80:20). Yield: 5.94 g (73% of theory).
1H-NMR (600 MHz, DMSO-d6): 6 [ppm] = 9.69 (t, 1H), 7.52 (dd, 1H), 7.42 (dd,
1H), 7.19 (dd, 1H),
4.21-4.12 (m, 1H), 3.26 (dd, 1H), 3.04 (dd, 1H).
Example 2.8C
(2R)-2 -(4-Chloro -3 -fluorobenzyl) -3,3,3 -trifluoropropan-1 -ol (single
stereoisomer)
CI F F
0 H
(2R)-2-(4-Chloro-3-fluorobenzy1)-3,3,3-trifluoropropanal (single stereoisomer)
(3.00 g, 11.8 mmol,
1.0 eq.) was dissolved in dichloromethane (21 ml) and cooled to -78 C.
Subsequently, the mixture
was treated with sodium borohydride (5.83 g, 58.9 mmol, 5.0 eq.) and precooled
methanol (10 ml, -
78 C). Stirring was continued at -78 C for 1 h, followed by the addition of a
saturated aqueous
ammonium chloride solution and warming to RT. The aqueous phase was then
extracted with
.. dichloromethane. The combined organic phases were dried over anhydrous
sodium sulfate, filtered
and concentrated under reduced pressure. The crude mixture was purified by
preparative HPLC
(reversed phase, eluent: water with 0.05% formic acid! acetonitrile 90:10 to
5:95). Yield: 1.86 g
(60% of theory).
1H-NMR (600 MHz, DM50-d6): 6 [ppm] = 7.51 (dd, 1H), 7.37 (dd, 1H), 7.17 (dd,
1H), 4.96 (t, 1H),
3.57-3.52 (m, 1H), 3.50-3.40 (m, 1H), 2.92-2.84 (m, 2H), 2.77-2.64 (m, 1H).
Example 2.9A
3 -(3 ,4 -Dichlorophenyl)propanal
CI
CI
0
1,2-Dichloro-4-iodobenzene (5.00 g, 18.0 mmol, 1.0 eq.), tetra-N-butylammonium
chloride (4.99 g,
18.0 mmol, 1.0 eq.) and sodium bicarbonate (3.77 g, 44.9 mmol, 2.5 eq.) were
dissolved in N,N-
dimethylformamide (30 ml) and argon was passed through the resulting
suspension for 10 min. After
addition of palladium(II) acetate (80.6 mg, 359 umol, 0.02 eq.) and prop-2-en-
1-ol (1.8 ml, 27 mmol,
1.5 eq.), the mixture was heated to 50 C and stirred at this temperature for 5
h, followed by the
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addition of methyl tert-butyl ether and water. After phase separation, the
aqueous phase was extracted
with methyl tert-butyl ether. The combined organic phases were washed with
water and brine, dried
over anhydrous sodium sulfate, filtered and concentrated under reduced
pressure. The residue was
purified by column chromatography (silica gel, eluent: cyclohexane / ethyl
acetate 80:20). Yield:
2.56 g (69% of theory).
1H-NMR (600 MHz, DMSO-d6): 6 [ppm] = 9.69 (t, 1H), 7.53-7.50 (m, 2H), 7.23
(dd, 1H), 2.86-2.79
(m, 4H).
Example 2.9B
(2R)-2-(3 ,4-Dichlorobenzy1)-3 ,3 ,3 -trifluoropropanal (single stereoisomer)
CI F F
CI
0
Argon was passed through a mixture of 3,3-dimethy1-1-(trifluoromethyl)-1,2-
benziodoxole (4.06 g,
12.3 mmol, 1.0 eq.), (5 R)-5-benzy1-2,2,3-trimethy1-4-oxoimidazolidin-l-ium
trifluoroacetate (single
stereoisomer) (818 mg, 2.46 mmol, 0.2 eq.) and copper(I) chloride (239 mg,
2.41 mmol, 0.075 eq.)
for 10 min. The mixture was cooled to -20 C before a solution of 3-(3,4-
dichlorophenyl)propanal
(2.55 g, 12.3 mmol, 1.0 eq.) in chloroform (35 ml) was added. The resulting
mixture was stirred
at -20 C for 5 h, kept in the freezer at -18 C overnight and stirred at -20 C
for additional 3 h. The
mixture was then immediately purified by column chromatography (silica gel,
eluent:
dichloromethane / methanol 98:2 to 80:20). Yield: 2.20 g (68% of theory).
Example 2.9C
(2R)-2-(3 ,4-Dichlorobenzy1)-3 ,3 ,3 -trifluoropropan-1 -ol (single
stereoisomer)
CI F F
0 H
CI
(2R)-2-(3,4-Dichlorobenzy1)-3,3,3-trifluoropropanal (single stereoisomer)
(2.20 g, 8.12 mmol,
1.0 eq.) was dissolved in dichloromethane (20 ml) and cooled to -78 C.
Subsequently, the mixture
was treated with sodium borohydride (4.02 g, 40.6 mmol, 5.0 eq.) and precooled
methanol (5.0 ml,
-78 C). Stirring was continued at -78 C for 1.5 h, followed by the addition of
saturated aqueous
ammonium chloride solution and warming to RT. The aqueous phase was then
extracted with
dichloromethane. The combined organic phases were dried over anhydrous sodium
sulfate, filtered
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and concentrated under reduced pressure. The crude mixture was used in the
subsequent step without
further purification. Yield: 1.82 g (62% purity, 51% of theory).
11-1-NMR (400 MHz, DMSO-d6): 6 [ppm] = 7.62-7.55 (m, 2H), 7.33-7.26 (m, 1H),
4.98 (t, 1H), 3.58-
3.43 (m, 2H), 2.91-2.84 (m, 2H), 2.78-2.66 (m, 1H).
Example 2.10A
(2Z)-3-(4-Chloropheny1)-2-(trifluoromethyl)prop-2-enoic acid
F 0
F OH
CI
Under nitrogen atmosphere, titanium tetrachloride (470 g, 2.48 mol) in
dichloromethane (200 ml)
was added dropwise at 0 C to a stirred mixture of 3,3,3-trifluoropropanoic
acid (210 g, 1.64 mol)
and 4-chlorobenzaldehyde (251 g, 1.79 mol) in tetrahydrofuran (11). The
resulting mixture was
stirred for 1 h at 0 C under nitrogen atmosphere, followed by the dropwise
addition of triethylamine
(684 ml, 4.92 mol) over 1 h. The resulting mixture was stirred for additional
12 h at RT. The reaction
was repeated in parallel for another 4 batches. The reaction mixtures were
separately quenched with
ice/water (5 1) and the resulting mixtures were combined and extracted three
times with
dichloromethane (5 1). The combined organic layers were washed two times with
saturated aqueous
ammonium chloride solution (5 1), dried over anhydrous sodium sulfate,
filtered and concentrated
under reduced pressure. The residue was slurried with petroleum ether (500
m1). The precipitated
solids were collected by filtration, washed with petroleum ether (500 ml) and
dried in vacuo . Yield:
1070 g (52% of theory).
LC-MS (method 19): Rt = 1.01 min; MS (ES): m/z = 249 [M-E1]
11-1-NMR (400 MHz, CDC13): 6 [ppm] = 11.92 (br s, 1H), 8.22 (s, 1H), 7.47-7.39
(m, 4H).
19F-NMR (376 MHz, CDC13): 6 [ppm] = -58.2 (s, 3F).
Example 2.10B
(2R)-2-(4-Chlorobenzy1)-3,3,3-trifluoropropanoic acid (single stereoisomer)
F 0
OH
CI
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(S)-2,6-Diethyldinaphtho [2,1-d: 1',2'-f] [1,3,21 dioxapho sphepine 4-oxide
(20.9 g, 53.9 mmol,
0.010 eq.) [Y. Li etal., Angew. Chem. Int. Ed. 2013, 10], 6748-67521 in 2-
propanol / water (4:1,
4.5 1) was treated at RT with bis(1,5-cyclooctadiene)rhodium(I)
tetrafluoroborate (10.9 g,
26.9 mmol, 0.015 eq.) and stirred for 2 min. Afterwards, triphenyl phosphine
(4.71 g, 18.0 mmol,
0.010 eq.) was added and stirring was continued for another 10 min. (2Z)-3-(4-
chloropheny1)-2-
(trifluoromethyl)prop-2-enoic acid (450 g, 1.80 mol) and morpholine (15.7 ml,
180 mmol, 0.10 eq.)
were added and the reaction mixture was transferred to a 7 1 autoclave.
Stirring was continued at a
hydrogen pressure of 20 bar for 24 h at RT. The reaction mixture was filtered
over diatomaceous
earth and the 2-propanol was mostly removed under reduced pressure. The
residue was treated with
1 N aqueous hydrogen chloride solution (1.0 1) and water (1.0 1). The mixture
was extracted with
ethyl acetate (4.5 1) and after phase separation the aqueous phase was
extracted with ethyl acetate (3
x 2.5 1). The combined organic phases were dried over sodium sulfate, filtered
and concentrated
under reduced pressure. The crude material was used in the subsequent reaction
without further
purification. Yield: 482 g (94% purity, quantitative of theory, >97% ee).
LC-MS (method 17): Rt = 6.82 min; MS (ESIpos): m/z = 253 [M+I-11+
Chiral SFC: Rt = 0.87 min, >97% ee
Analysis method: SFC: column: Chiralpak AD-3, 3 um, 100 mm x 4.6 mm, eluent:
isocratic 95%
carbon dioxide / 5% methanol, 10 min, temperature: 40 C; flow rate: 3.0
ml/min; UV detection:
210 nm, backpressure: 130 bar.
1H-NMR (400 MHz, DM50-d6): 6 [ppm] = 13.34 (br s, 1H), 7.46-7.19 (m, 4H), 3.79
(td, 1H), 3.14-
2.97 (m, 2H).
Example 2.10C
Methyl (2R)-2-(4-chlorobenzy1)-3,3,3-trifluoropropanoate (single stereoisomer)
F 0
CH3
0'
CI
Methanol (3.6 1) was treated dropwise with thionyl chloride (346 ml, 4.75 mol,
2.0 eq.) at -10 C.
After stirring for another 10 min at 0 C, (2R)-2-(4-chlorobenzy1)-3,3,3-
trifluoropropanoic acid
(single stereoisomer) (600 g, 2.38 mol) was added and the reaction mixture was
stirred for 3 h under
reflux. The reaction mixture was concentrated under reduced pressure and the
residue was
redissolved in ethyl acetate (4.0 1). The organic phase was washed with
saturated aqueous sodium
bicarbonate (3.0 1) and 10% aqueous sodium chloride solution (2.5 1), dried
over anhydrous sodium
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sulfate, filtered and concentrated under reduced pressure. The residue was
redissolved in
dichloromethane, treated with silica gel (3.0 kg) and the mixture was
concentrated under reduced
pressure. This material was purified by flash column chromatography (9.0 kg
silica gel, eluent:
petroleum ether! dichloromethane 8:2). Yield: 493 g (78% of theory).
GC-MS (method 2): Rt = 3.91 min; MS (EIpos): m/z = 266 [M]
1H-NMR (600 MHz, DM50-c/6): 6 [ppm] = 7.40-7.34 (m, 2H), 7.33-7.28 (m, 2H),
4.07-3.96 (m,
1H), 3.61 (s, 3H), 3.18-3.05 (m, 2H).
Example 2.10D
(2R)-2-(4-Chlorobenzy1)-3,3,3-trifluoropropan-1-ol (single stere oi some r)
OH
C
I
Two experiments of the same scale were run in parallel; the purification was
carried out with the
combined reaction mixtures.
Methyl (2R)-2-(4-chlorobenzy1)-3,3,3-trifluoropropanoate (single stereoisomer)
(171 g, 641 mmol)
in tetrahydrofuran (3.2 1) was treated under argon with lithium aluminium
hydride (673 ml, 2.4 M
solution in tetrahydrofuran, 673 ml, 1.62 mol, 2.52 eq.) at -10 C within 10
min. Afterwards, the
reaction mixture was stirred while allowing to warm up to RT. The reaction
mixture was diluted with
tert-butylmethyl ether (700 ml), cooled to 0 C and carefully treated dropwise
with water (61.4 ml)
under stirring. Afterwards, the reaction mixture was carefully treated
dropwise with 15% aqueous
sodium hydroxide solution (61.4 ml) and finally again with water (184 m1). The
reaction mixture was
warmed to RT, stirred for 15 min and then treated with anhydrous magnesium
sulfate (1.4 kg). After
stirring for another 15 min, the reaction mixture was filtered under reduced
pressure and the filter
cake was washed with tert-butylmethyl ether (1.7 1). The organic phase was
concentrated under
reduced pressure to yield the crude material. The combined crude material from
two batches was
subjected to column chromatography (8.0 kg silica gel, eluent:
dichloromethane). Yield for the
combined two batches: 271.4 g (89% of theory, >95% ee).
GC-MS (method 2): Rt = 4.23 min; MS (EIpos): m/z = 238 [M]
Chiral SFC: Rt = 0.85 min, >95% ee
Analysis method: SFC: column: Chiralcel OD-3, 3 lam, 100 mm x 4.6 mm, eluent:
isocratic 90%
carbon dioxide! 10% isopropanol, 10 min, temperature: 40 C; flow rate: 3.0
ml/min; UV detection:
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210 nm, backpressure: 130 bar.
1H-NMR (600 MHz, DMSO-d6): 6 [ppm] = 7.39-7.34 (m, 2H), 7.33-7.29 (m, 2H),
5.00 (t, 1H), 3.61-
3.45 (m, 2H), 2.93-2.81 (m, 2H), 2.63 (ddt, 1H).
Example 2.11A
2-(4-Chlorobenzy1)-3-hydroxypropanenitrile (racemate)
CI
CN
OH
Lithium bis(trimethylsilyl)amide (1.41 1, 1.0 M in tetrahydrofuran, 1.41 mol,
2.0 eq.) was added
at -78 C to solution of 3-hydroxypropanenitrile (50.0 g, 704 mmol) in
tetrahydrofuran (500 ml) and
the reaction mixture was stirred at -78 C for 1 h, followed by addition of 1-
(bromomethyl)-4-
chlorobenzene (145 g, 704 mmol, 1.0 eq.) in tetrahydrofuran (200 ml) at -78 C.
The reaction mixture
was stirred at -78 C for 1 h and then at 0 C for 1 h. The mixture was diluted
with saturated aqueous
solution of ammonium chloride and extracted with ethyl acetate. The combined
organic layers were
washed with brine, dried over anhydrous sodium sulfate, filtered and
concentrated under reduced
pressure. The residue was purified by column chromatography (silica gel,
eluent: petroleum ether /
ethyl acetate 20:1 to 10:1). Yield: 55.0 g (90% purity, 36% of theory).
1H-NMR (400 MHz, CDC13): 6 [ppm] = 7.16 (d, 2H), 7.07-7.02 (m, 2H), 3.61 (q,
2H), 2.79 (s, 3H),
2.49-2.42 (m, 1H).
Example 2.11B
3 -{ [tert-Butyl(diphenyOsilyll oxy} -2-(4-chlorobenzyl)propanenitrile
(racemate)
CI
NC
0
'Si CH3
CHC3H3
tert-Butyl(chloro)diphenylsilane (72.2 ml, 77.3 g, 281 mmol, 1.0 eq.) was
added at RT to a solution
of 2-(4-chlorobenzy1)-3-hydroxypropanenitrile (racemate) (55.0 g, 90% purity,
252 mmol), 4-
dimethylaminopyridine (1.72 g, 14.1 mmol, 0.055 eq.) and imidazole (57.4 g,
843.4 mmol, 3.3 eq.)
in dichloromethane (750 m1). The reaction mixture was stirred at RT for 12 h.
Subsequently, the
mixture was diluted with water and extracted with dichloromethane. The
combined organic layers
were washed with brine, dried over anhydrous sodium sulfate, filtered and
concentrated under
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reduced pressure. The residue was purified by column chromatography (silica
gel, eluent: petroleum
ether! ethyl acetate 20:1 to 10:1). Yield: 75.0 g (87% purity, 59% of theory).
1H-NMR (400 MHz, CDC13): 6 [ppm] = 7.60-7.53 (m, 4H), 7.37-7.27 (m, 6H), 7.18
(d, 2H), 7.04 (d,
2H), 3.66 (d, 2H), 2.92-2.83 (m, 2H), 2.83-2.73 (m, 1H), 1.02 (s, 9H).
Example 2.11C
3 -{ [tert-Butyl(diphenyOsilyll oxy} -2-(4-chlorobenzyl)propanal (racemate)
CI 0 H
0
C H3
* hCH3
C H3
A solution of diisobutylaluminium hydride (618 ml, 1.0 M in toluene, 4.6 eq.)
was added at -70 C to
a solution of 3- { [tert-butyl(diphenyl)silylloxy} -2-(4-
chlorobenzyl)propanenitrile (racemate) (67.0 g,
87% purity, 133 mmol) in toluene (1.0 1) and the reaction mixture was stirred
at 0 C for 2 h. The
mixture was diluted with saturated potassium sodium tartrate solution and
extracted with ethyl
acetate. The combined organic layers were washed with brine, dried over
anhydrous sodium sulfate,
filtered and concentrated under reduced pressure. The crude product was used
in the subsequent
reaction without further purification. Yield: 80.0 g.
Example 2.11D
tert-Butyl [2 -(4-chlorobenzy1)-3 ,3 -difluoropropoxy] diphenylsilane
(racemate)
CI F F
0
C H3
= hC H3
CH3
(Diethylamino)sulfur trifluoride (72.6 ml, 88.5 g, 549 mmol, 3.0 eq.) was
added at -70 C to a solution
of crude 3-{[tert-butyl(diphenyl)silylloxy}-2-(4-chlorobenzyl)propanal
(racemate) (80.0 g, 183
mmol) in dichloromethane (1.0 1) and the reaction mixture was stirred at RT
for 2 h. The mixture
was diluted with water and extracted with dichloromethane. The combined
organic layers were
washed with brine, dried over anhydrous sodium sulfate, filtered and
concentrated under reduced
pressure. The residue was purified by column chromatography (silica gel,
eluent: petroleum ether!
ethyl acetate 100:0 to 20:1). Yield: 17.0 g (65% purity, 18% of theory over
two steps).
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1H-NMR (400 MHz, CDC13): 6 [ppm] = 7.63-7.55 (m, 4H), 7.48-7.37 (m, 6H), 7.22-
7.17 (m, 2H),
7.02 (d, 2H), 6.05 (d, 1H), 3.74-3.67 (m, 1H), 3.64-3.58 (m, 1H), 2.98-2.84
(m, 1H), 2.78-2.69 (m,
1H), 2.32-2.11 (m, 1H), 1.11-1.08 (m, 9H).
Example 2.11E
2-(4-Chlorobenzy1)-3,3-difluoropropan-1-ol (racemate)
CL F F
0 H
A solution of tetra-N-butylammonium fluoride (74.1 ml, 1.0 M in
tetrahydrofuran, 74.1 mmol,
3.1 eq.) was added at -30 C
to solution of te rt-butyl [2-(4-chlorobenzy1)-3,3-
difluoropropoxyldiphenylsilane (racemate) (17.0 g, 65% purity, 24.1 mmol) in
tetrahydrofuran (180
ml) and the reaction mixture was stirred at RT for 2 h. The mixture was
filtered and concentrated
under reduced pressure to give a residue which was dissolved in ethyl acetate.
The combined organic
layers were washed with brine, dried over anhydrous sodium sulfate, filtered
and concentrated under
reduced pressure. The crude mixture was purified by preparative HPLC. Yield:
5.50 g (99% of
theory).
1H-NMR (400 MHz, CDC13): 6 [ppm] = 7.34-7.28 (m, 2H), 7.17 (d, 2H), 6.35-5.56
(m, 1H), 3.89-
3.59 (m, 2H), 3.13-2.56 (m, 2H), 2.45-2.13 (m, 1H), 1.55 (br s, 1H).
Example 3.1A
2-Methoxy-5 -(methoxymethoxy)-4-(4,4,5,5-tetramethy1-1,3,2-dioxaborolan-2-
yl)pyridine
CH3
0
3C 0 CL0 CH3
'
H3C 0
H3C
CH3
A solution of tert-butyllithium (22.2 ml, 1.6 M in pentane, 35.4 mmol, 2.0
eq.) was added at -78 C
under nitrogen atmosphere to a solution of 2-methoxy-5-
(methoxymethoxy)pyridine (3.0 g, 17.7
mmol, 1.0 eq.) in tetrahydrofuran (60 m1). After stirring at -78 C for 0.5 h,
2-isopropoxy-4,4,5,5-
tetramethy1-1,3,2-dioxaborolane (3.96 g, 21.3 mmol, 1.2 eq.) was added
dropwise. The reaction
mixture was stirred at -78 C for 2 h, quenched with methanol at -78 C and then
concentrated under
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reduced pressure. The crude product was used in the subsequent step without
further purification.
Yield: 5.25 g.
Example 3.1B
3 - {4 -Chloro -242-methoxy-5 -(methoxymethoxy)pyridin-4-yll phenyl} -2 -
methylpropan-1 -ol
(racemate)
0
H 3C'
0
N
CI
0 H
H 3C
2-Methoxy-5 -(methoxymethoxy)-4-(4,4,5,5-tetramethy1-1,3,2-dioxaborolan-2-
yl)pyridine (crude
product from previous step, 7.39 g, 25.0 mmol, 2.0 eq.), 1,1'-
bis(diphenylphosphino)ferrocene-
palladium(II) chloride (0.92 g, 1.3 mmol, 0.1 eq.) and sodium carbonate (3.98
g, 37.6 mmol, 3.0 eq.)
were added under nitrogen atmosphere to a solution of 3-(2-bromo-4-
chloropheny1)-2-
methylpropan-1-ol (racemate) (3.3 g, 12.5 mmol, 1.0 eq.) in 1,4-dioxane (100
ml) and water (15 m1).
The resulting mixture was stirred at 90 C for 2 h, cooled to RT and
concentrated under reduced
pressure. The residue was purified by column chromatography (silica gel,
eluent: petroleum ether!
ethyl acetate 1:1). Yield: 3.6 g (87% purity, 72% of theory).
LC-MS (method 11): Rt = 1.14 min; MS (ESIpos): m/z = 352 [M+F11+
Example 3.1C
445 -Chloro-2 -(3 -hydroxy-2-methylpropyl)phenyl] -6 -methoxypyridin-3 -ol
(racemate)
HO
N
CI
o-C H 3
H 3C OH
Aqueous hydrochloric acid (6.0 ml, 3 N) was added to a solution of 3-{4-chloro-
242-methoxy-5-
(methoxymethoxy)pyridin-4-yllpheny11-2-methylpropan-1-ol (racemate) (2.2 g,
87% purity,
5.4 mmol, 1.0 eq.) in tetrahydrofuran (30 m1). The reaction mixture was
stirred at 60 C for 3 h and
concentrated under reduced pressure. The residue was purified by column
chromatography (silica
gel, eluent: petroleum ether! ethyl acetate 1:1). Yield: 1.60 g (83% purity,
80% of theory).
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LC-MS (method 12): Rt = 0.79 min; MS (ESIpos): m/z = 308 [M+I-11+
1H-NMR (300 MHz, DM50-d6): 6 [ppm] = 9.33 (s, 1H), 7.78 (s, 1H), 7.39-7.35 (m,
1H), 7.30-7.27
(m, 1H), 7.13 (d, 1H), 6.54 (s, 1H), 4.48-4.31 (br s, 1H), 3.77 (s, 3H), 3.15-
2.98 (m, 2H), 2.63-2.56
(m, 1H), 2.19-2.11 (m, 1H), 1.61-1.52 (m, 1H), 0.60 (d, 3H).
Example 3.1D
543 -(2 -B romo -4-chloropheny1)-2 -methylpropoxy] -4 -iodo-2-methoxypyridine
(racemate)
CI
C H 3
0;a
Br I ,CH3
I 0
3-(2-Bromo-4-chloropheny1)-2-methylpropan-1-ol (racemate) (2.0 g, 7.2 mmol,
1.0 eq.) and
(tributylphosphoranylidene)acetonitrile (2.62 g, 10.8 mmol, 1.5 eq.) were
added at RT to a
suspension of 4-iodo-6-methoxypyridin-3-ol (1.82 g, 7.2 mmol, 1.0 eq.) in
toluene (10 m1). The
reaction mixture was irradiated at 160 C for 2 h in a microwave reactor and
then cooled to RT. The
above reaction procedure was repeated for three more batches (altogether 8.02
g (28.9 mmol) 3-(2-
Bromo-4-chloropheny1)-2-methylpropan-1-ol (racemate)). The four batches were
combined and
concentrated under reduced pressure. The residue was diluted with ethyl
acetate and washed with
water and brine. The organic layer was dried over anhydrous sodium sulfate,
filtered and
concentrated under reduced pressure. The residue was purified by column
chromatography (silica
gel, eluent: petroleum ether! ethyl acetate 19:1). Yield: 11.87 g (81% of
theory).
LC-MS (method 13): Rt = 1.63 min; MS (ESIpos): m/z = 496 [M+I-11+
1H-NMR (400 MHz, DMSO-d6): 6 [ppm] = 7.77-7.70 (m, 2H), 7.42 (dd, 1H), 7.38-
7.30 (m, 2H),
3.98-3.86 (m, 2H), 3.78 (s, 3H), 3.08-2.99 (m, 1H), 2.73-2.63 (m, 1H), 2.35-
2.20 (m, 1H), 1.02 (d,
3H).
Example 3.1E
11-Chloro-2-methoxy-7-methyl-7, 8-dihydro-6H- [3] benzoxocino [2,1-c] pyridine
(racemate)
H 3C 0
N
C H 3
=
C
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Diisopropyl diazene-1,2-dicarboxylate (2.1 g, 10.4 mmol, 2.4 eq.) was added
dropwise at 0 C to a
suspension of 445 -chloro-2-(3 -hydroxy-2-methylpropyl)phenyl] -6 -
methoxypyridin-3 -ol (racemate)
(1.6 g, 83% purity, 4.4 mmol) and triphenylphosphine (2.7 g, 10.4 mmol, 2.4
eq.) in tetrahydrofuran
(80 m1). The resulting mixture was stirred under nitrogen atmosphere at RT for
3 h and then
.. concentrated under reduced pressure. The residue was purified by column
chromatography (silica
gel, eluent: petroleum ether! ethyl acetate 10:1). Yield: 0.6 g (91% purity,
43% of theory).
LC-MS (method 12): Rt = 1.27 min; MS (ESIpos): m/z = 290 [M+I-11+
1H-NMR (300 MHz, DM50-d6): 6 [ppm] = 8.05 / 8.03 (2s, 1H), 7.47-7.31 (m, 3H),
6.78 / 6.73 (2s,
1H), 4.49-4.43 (m, 1H), 3.86 (s, 3H), 3.45-3.38 (m, 1H), 2.70-2.57 (m, 1H),
2.21-2.13 (m, 1H), 2.03-
1.96 (m, 1H), 0.90-0.83 (d, 3H). Additional signals of minor rotamers were
also detected.
Alternative synthetic route:
Bis(pinacolato)diboron (6.14 g, 24.2 mmol, 2.0 eq.), palladium(II) acetate
(0.27 g, 1.2 mmol, 0.1
eq.), tricyclohexylphosphine (0.51 g, 1.8 mmol, 0.15 eq.) and potassium
acetate (3.56 g, 36.2 mmol,
3.0 eq.) were added to a solution of 443-(2-bromo-4-chloropheny1)-2-
methylpropoxy1-5-iodo-2-
methoxypyridine (racemate) (6.00 g, 12.1 mmol, 1.0 eq.) in /V,N-
dimethylformamide (60 m1). The
resulting mixture was purged with nitrogen for 5 min, stirred at 80 C for 15 h
under nitrogen
atmosphere, treated with 5 ml of saturated aqueous solution of sodium
bicarbonate and stirred at
80 C for further 2 h. After cooling to RT, the reaction mixture was diluted
with ethyl acetate and
filtered through Celite . The filtrate was washed with water and brine, dried
over anhydrous sodium
sulfate, filtered and concentrated under reduced pressure. The residue was
purified by column
chromatography (silica gel, eluent: petroleum ether! ethyl acetate 19:1).
Yield: 1.2 g (34% of theory).
LC-MS (method 13): Rt = 1.38 min; MS (ESIpos): m/z = 290 [M+I-11+
Example 3.1F
11-Chloro-7-methyl-7,8-dihydro-3H-[3]benzoxocino [2,1-c] pyridin-2 (6H)-one
(racemate)
0
H 3C NH
0
C
I
4-Toluenesulfonic acid monohydrate (2.72 g, 14.3 mmol, 1.8 eq.) and lithium
iodide (5.32 g, 39.7
mmol, 5.0 eq.) were added at RT to a solution of 11-chloro-2-methoxy-7-methy1-
7,8-dihydro-6H-
[3]benzoxocino[2,1-clpyridine (racemate) (2.30 g, 7.9 mmol) in 1-butanol (20
m1). After stirring at
85 C for 12 h, the mixture was concentrated under reduced pressure. The
residue was dissolved in
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dichloromethane, washed with water and brine. The organic layer was dried over
anhydrous sodium
sulfate, filtered and concentrated under reduced pressure. The residue was
purified by column
chromatography (silica gel, eluent: dichloromethane / methane 19:1). Yield:
1.78 g (79% of theory).
LC-MS (method 14): Rt = 1.21 min; MS (ESIpos): m/z = 276 [M+I-11+
1H-NMR (400 MHz, DMSO-d6): 6 [ppm] = 11.56-11.25 (br s, 1H), 7.50-7.36 (m,
3H), 7.34-7.27 (m,
1H), 6.33-6.25 (m, 1H), 4.45-4.35 / 4.23-4.16 (2m, 1H), 3.86-3.78 / 3.3-3.25
(2m, 1H, partially
concealed), 2.86-2.78 / 2.72-2.59 (2m, 1H), 2.29 (dd, 1H), 2.07-1.90 (m, 1H),
0.94-0.80 (m, 3H).
Additional signals of minor rotamers were also detected.
Example 3.1G
(7 R) - 11 -Chloro-7-methy1-7,8-dihydro-3H- [3] benzoxocino [2,1-c] pyridin-2
(6H)-one (single
stereoisomer)
0
H 3C NH
0
CI
Enantiomer separation of 1.74 g of 11-chloro-7-methy1-7,8-dihydro-3H-
[3]benzoxocino[2,1-
c]pyridin-2(6H)-one (racemate), Example 3.1F gave
single stereoisomer 1 (the title compound Example 3.1G) (chiral HPLC: R1 =
3.00 min, >99% ee):
603 mg (86% purity),
single stereoisomer 2 (chiral HPLC: R1 = 5.79 min): 552 mg.
Separation method: HPLC: column: Daicel Chiralpak AS-H 5 lam, 250 mm x 20 mm;
eluent: 15%
n-heptane / 85% ethanol; temperature: 40 C; flow rate: 20 ml/min; UV
detection: 220 nm.
Analysis method: HPLC: column: Daicel Chiralpak AS-3 3 lam, 50 mm x 4.6 mm;
eluent: 50%
n-heptane / 50% ethanol; temperature: 40 C; flow rate: 1 ml/min; UV detection:
220 nm.
LC-MS (method 4): R1= 1.52 min; MS (ESIpos): m/z = 276 [M+I-11+
Example 3.2A
543 -(2 -B romo -4-chloro -3 -fluoropheny1)-2-methylpropoxy] -4-iodo-2 -
methoxypyridine (racemate)
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CI
C H 3
o;oNL
Br I C H 3
General Method 2 was carried out three times with 3-(2-bromo-4-chloro-3-
fluoropheny1)-2-
methylpropan-1-ol (racemate) (500 mg, 1.78 mmol, 1.0 eq.), 4-iodo-6-
methoxypyridin-3-ol (446 mg,
1.78 mmol, 1.0 eq.) and (tributylphosphoranylidene)acetonitrile (643 mg, 2.66
mmol, 1.5 eq.) in
toluene (14 ml) at 120 C including the following variations of the procedure:
The three reaction
mixtures were combined, concentrated under reduced pressure and the crude
mixture was purified
by column chromatography (silica gel, eluent: cyclohexane / ethyl acetate 96:4
to 70:30). Yield: 2.01
g (92% purity, 68% of theory).
LC-MS (method 3): R1= 5.21 min; MS (ESIpos): m/z = 516 [M+1-11+
Example 3.2B
11 -Chloro-12-fluoro -2 -methoxy-7 -methy1-7,8 -dihydro -6H- [3] benzoxocino
[2,1 -c] pyridine
(racemate)
0
H 3C N
C H3
* F
CI
General Method 3a was carried out two times with 5 43-(2-bromo-4-chloro-3-
fluoropheny1)-2-
methylpropoxy1-4-iodo-2-methoxypyridine (racemate) (1.00 g, 92% purity, 1.79
mmol, 1.0 eq.),
bis(pinacolato)diboron (636 mg, 2.50 mmol, 1.4 eq.), potassium acetate (526
mg, 5.36 mmol, 3.0
eq.), palladium(II) acetate (20.0 mg, 89.0 umol, 0.05 eq.) and
tricyclohexylphosphine (50.1 mg, 179
umol, 0.1 eq.) in N,N-dimethylformamide (26 ml) including the following
variations of the
procedure: After stirring at 80 C overnight,
tetrakis(triphenylphosphine)palladium(0) (103.7 mg,
89.0 umol, 0.05 eq.) and saturated aqueous sodium bicarbonate solution (12.5
ml) were added to
each of the two reaction mixtures. Stirring was then continued at 85 C for 1
h, the two reaction
mixtures were combined, water was added and the combined mixtures were
extracted with ethyl
acetate, dried over anhydrous sodium sulfate, filtered and concentrated under
reduced pressure. The
crude mixture was purified by preparative HPLC (reversed phase, eluent: water
with 0.05% formic
acid! acetonitrile 90:10 to 5:95). Yield: 485 mg (84% purity, 37% of theory).
LC-MS (method 3): Rt = 3.89 min; MS (ESIpos): m/z = 308 [M+1-11+
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Example 3.2C
11-Chloro-12-fluoro -7 -methy1-7, 8-dihydro-3H- [3] benzoxocino [2,1-c]
pyridin-2 (6H)-one (racemate)
0
H 3C NH
0
* F
CI
General Method 4 was carried with 11-chloro-12-fluoro-2-methoxy-7-methy1-7,8-
dihydro-6H-
[31benzoxocino[2,1-clpyridine (racemate) (480 mg, 84% purity, 1.31 mmol, 1.0
eq.) and sodium
iodine (393 mg, 2.62 mmol, 2.0 eq.) in acetic acid (15 ml) at 100 C for 4 h
including the following
variations of the procedure: The crude mixture was purified by preparative
column chromatography
(silica gel, eluent: dichloromethane / methanol 100:0 to 90:10), followed by
purification by
preparative HPLC (reversed phase, eluent: water with 0.05% formic acid /
acetonitrile 90:10 to 5:95).
Yield: 200 mg (92% purity, 48% of theory).
LC-MS (method 3): Rt = 2.37 min; MS (ESIpos): m/z = 294 [M+H1+
1H-NMR (500 MHz, DM50-c/6): 6 [ppm] = 11.49 (br s, 1H), 7.70-7.47 (m, 2H),
7.30-7.17 (m, 1H),
6.40 / 6.37 (2d, 1H), 4.44-4.36 / 3.84-3.79 (2m, 1H), 4.24-4.19 / 3.33-3.28
(2m, 1H), 2.74-2.66 /
2.72-2.67 (2m, 1H), 2.31-2.18 (m, 1H), 2.05-1.88 (m, 1H), 0.88 / 0.81 (2d,
3H).
Example 3.3A
5-[2-(2-Bromo-4-chlorobenzyl)butoxy]-4-iodo-2-methoxypyridine (racemate)
CI *I 3C
0
I "
Br CH3
I 0'
General Method 2 was carried out two times with 2-(2-bromo-4-
chlorobenzyl)butan-1-ol (racemate)
(500 mg, 77% purity, 1.39 mmol, 1.0 eq.), 4-iodo-6-methoxypyridin-3-ol (453
mg, 1.80 mmol, 1.3
eq.) and (tributylphosphoranylidene)acetonitrile (502 mg, 2.08 mmol, 1.5 eq.)
in toluene (15 ml) at
120 C including the following variations of the procedure: The two reaction
mixtures were combined
and purified by column chromatography (silica gel, eluent: cyclohexane / ethyl
acetate 100:0 to
65:35). Yield: 750 mg (63% purity, 33% of theory).
LC-MS (method 1): R1= 1.62 min; MS (ESIpos): m/z = 510 [M+H1+
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Example 3.3B
11 -Chloro-7-ethy1-2 -methoxy-7,8 -dihydro -6H-[3] benzoxocino [2,1 -c]
pyridine (racemate)
0
N
H3C
CH-2
CI
General Method 3a was carried out with 542-(2-bromo-4-chlorobenzyl)butoxy1-4-
iodo-2-
methoxypyridine (racemate) (750 mg, 63% purity, 925 ma 1.0 eq.),
bis(pinacolato)diboron (352
mg, 1.39 mmol, 1.5 eq.), palladium(II) acetate (10.4 mg, 46.3 ma 0.1 eq.),
tricyclohexylphosphine
(26.0 mg, 92.5 urnol, 0.1 eq.) and potassium acetate (272 mg, 2.78 mmol, 3.0
eq.) in N,N-
dimethylformamide (14 ml) including the following variations of the procedure:
After stirring at
80 C overnight, tetrakis(triphenylphosphine)palladium(0) (53.7 mg, 46.3 urnol,
0.1 eq.) and
saturated aqueous sodium bicarbonate solution (6.5 ml) were added to the
reaction mixture. Stirring
was then continued at 85 C for 3 days. The reaction mixture was diluted with
water and extracted
with ethyl acetate. The combined organic phases were dried over anhydrous
sodium sulfate, filtered
over silica gel (eluent: ethyl acetate) and used in the subsequent step
without further purification.
Yield: 600 mg (52% purity).
LC-MS (method 1): R1= 1.31 min; MS (ESIpos): m/z = 304 [M-411+
Example 3.3C
11 -Chloro-7-ethy1-7, 8-dihydro-3H- [3] benzoxocino [2, 1 -c] pyridin-2 (6H)-
one (racemate)
0
H 3C NH
CI
General Method 4 was carried out with 11-chloro-7-ethy1-2-methoxy-7,8-dihydro-
6H-
[3]benzoxocino[2,1-clpyridine (racemate) (600 mg, 52% purity, 1.03 mmol, 1.0
eq.) and sodium
iodide (462 mg, 3.08 mmol, 3.0 eq.) in acetic acid (15 ml) at 100 C for 2 h
including the following
variations of the procedure: The crude mixture was purified by preparative
HPLC (reversed phase,
eluent: water with 0.05% formic acid / acetonitrile 90:10 to 5:95). Yield: 57
mg (66% purity, 13%
of theory) and 15 mg (86% purity, 4% of theory).
LC-MS (method 1): R1 = 0.89 min; MS (ESIpos): m/z = 290 [M-411+
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Example 3.4A
-{[4 -(2 -B romo -4-chloropheny1)-3 -methylbutan-2 -yll oxy} -4 -iodo -2 -
methoxypyridine (mixture of
stereoisomers)
CI
CH 3
0 N
Br CH1a)ao-CH3
5 General Method 2 was carried out with 4-(2-bromo-4-chloropheny1)-3-
methylbutan-2-ol (mixture of
stereoisomers) (500 mg, 91% purity, 1.64 mmol, 1.0 eq.), 4-iodo-6-
methoxypyridin-3-ol (416 mg,
1.64 mmol, 1.0 eq.) and (tributylphosphoranylidene)acetonitrile (475 mg, 1.97
mmol, 1.2 eq.) in
toluene (15 ml) at 120 C including the following variations of the procedure:
After stirring for 1 h,
additional amounts of (tributylphosphoranylidene)acetonitrile (198 mg, 820
umol, 0.5 eq.) were
added and stirring was continued at 120 C for 1 h. The crude mixture was
purified by column
chromatography (silica gel, eluent: cyclohexane / ethyl acetate 98:2 to
80:20). Yield: 670 mg (78%
of theory).
LC-MS (method 1): R1= 1.61 min; MS (ESIpos): m/z = 510 1M+H1+
Example 3.4B
11 -Chloro-2-methoxy-6,7 -dimethy1-7, 8-dihydro-6H- [3] benzoxocino [2,1 -c]
pyridine (mixture of
stereoisomers)
H 3C
0
H 3C
N
o-CH 3
CI
General Method 3a was carried out with 54[4-(2-bromo-4-chloropheny1)-3-
methylbutan-2-ylloxyl-
4-iodo-2-methoxypyridine (mixture of stereoisomers) (670 mg, 1.29 mmol, 1.0
eq.),
bis(pinacolato)diboron (457 mg, 1.80 mmol, 1.4 eq.), palladium(II) acetate
(14.4 mg, 64.3 umol,
0.05 eq.) and potassium acetate (379 mg, 3.86 mmol, 3.0 eq.) in N,N-
dimethylformamide (16 ml)
including the following variations of the procedure: After stirring at 80 C
overnight,
tetrakis(triphenylphosphine)palladium(0) (74.6 mg, 64.3 umol, 0.05 eq.) and a
saturated aqueous
sodium bicarbonate solution (7.7 ml) were added to the reaction mixture.
Stirring was then continued
at 85 C overnight. The reaction mixture was filtered over a mixture of silica
gel and anhydrous
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sodium sulfate and eluted with a mixture (1:1) of dichloromethane and ethyl
acetate. The filtrate was
concentrated under reduced pressure, water was added and the combined mixture
was extracted with
ethyl acetate, dried over anhydrous sodium sulfate, filtered, concentrated
under reduced pressure and
used in the next step without further purification. Yield: 552 mg (63% purity,
89% of theory).
LC-MS (method 1): R1= 1.34 min; MS (ESIpos): m/z = 304 [M+F11+
Example 3.4C
11-Chloro-6,7-dimethy1-7,8-dihydro-3H431benzoxocinop,1-clpyridin-2(6H)-one
(mixture of
stereoisomers)
H 3C
0
H 3C
NH
0
CI
General Method 4 was carried out with 11-chloro-2-methoxy-6,7-dimethy1-7,8-
dihydro-6H-
Plbenzoxocino[2,1-clpyridine (mixture of stereoisomers) (550 mg, 63% purity,
1.14 mmol, 1.0 eq.)
and sodium iodide (513 mg, 3.42 mmol, 3.0 eq.) in acetic acid (20 ml) at 100 C
for 2 h including the
following variations of the procedure: The crude mixture was purified by
preparative HPLC
(reversed phase, eluent: water with 0.05% formic acid / acetonitrile 90:10 to
5:95). Yield: 157 mg
(75% purity, 36% of theory).
LC-MS (method 3): Rt = 2.52 min; MS (ESIpos): m/z = 290 [M-411+
Example 3.5A
5 -( { cis-24(2-Bromo -4 -chlorophenyl)methyll cyclobutylloxy)-4-iodo-2-
methoxypyridine
(racemate)
Br
C I C H
4-Iodo-6-methoxypyridin-3-ol (2.41 g, 9.62 mmol,
1.0 eq.), trans-24(2-bromo -4-
chlorophenyl)methyll -cyclobutan-l-ol (racemate) (2.65 g,
9.62 mmol) and
(tributylphosphoranylidene)-acetonitrile (2.52 ml, 9.62 mmol. 1.0 eq.) were
dissolved in toluene (53
ml) and equally distributed into three microwave vials which were sealed
afterwards. The reaction
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mixtures were heated at 120 C for 45 min under microwave irradiation. The
three mixtures were
combined and all volatiles removed in vacuo. The residue was dissolved in
ethyl acetate and washed
with water, dried over anhydrous sodium sulfate, filtered and concentrated
under reduced pressure.
The crude product was purified by column chromatography (silica gel, eluent:
cyclohexane / ethyl
acetate mixtures). Yield: 2.69 g (54% of theory).
LC-MS (method 4): Rt = 2.90 / 2.98 min; MS (ESIpos): m/z = 508 [M+F11+
Example 3.5B
cis-6-Chloro -3 -methoxy-9a,10,11,11a-tetrahydro-9H-cyc10but44,5] [3]
benzoxocino [2,1-c] pyridine
(racemate)
N
I C H 3
C
i
Bis(pinacolato)diboron (5.17 g, 20.35 mmol, 1.5 eq.), potassium acetate (3.99
g, 40.70 mmol,
3.0 eq.), palladium(II) acetate (305 mg, 1.36 mmol, 0.1 eq.) and
tricyclohexylphosphine (380 mg,
1.36 mmol, 0.1 eq.) were added to
a solution of 5-({cis-2-[(2-bromo-4-
chlorophenyl)methyllcyclobutyl}oxy)-4-iodo-2-methoxypyridine (racemate) (6.90
g, 13.57 mmol)
in NN-dimethylformamide (136 ml, degassed before in a flame-dried flask). The
reaction mixture
was stirred at 100 C overnight. Tetrakis(triphenylphosphine)palladium(0) (784
mg, 0.68 mmol, 0.05
eq.), solid sodium bicarbonate (5.70 g, 67.84 mmol, 5.0 eq.) and water (0.93
ml) were added. The
reaction mixture was stirred at 100 C overnight, allowed to come to RT,
diluted with ethyl acetate
and filtered through Celite . The filter cake was washed with ethyl acetate.
The combined filtrates
were concentrated under reduced pressure. The residue was dissolved ethyl
acetate, washed with
water, dried over anhydrous sodium sulfate, filtered and concentrated in vacuo
. The crude product
was used without further purification in the next step. Yield: 8.50 g.
LC-MS (method 4): Rt = 2.42 min; MS (ESIpos): m/z = 302 [M+I-11+
Example 3.5C
cis-6-Chloro-9a,10,11,11a-tetrahydro-2H-cyc10but44,5] [3] benzoxocino [2,1-c]
pyridin-3 (911)-one
(racemate)
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= 0
NH
0
CI
Sodium iodide (10.3 g, 68.8 mmol) was added to a solution of cis-6-chloro-3-
methoxy-9a,10,11,11a-
tetrahydro-9H-cyc1obut44,51[31benzoxocino[2,1-c]pyridine (racemate) (crude
product from
previous step, calculated with 50% purity: 8.3 g, 13.8 mmol) in acetic acid
(208 m1). The resulting
mixture was stirred at 80 C overnight. All volatiles were removed in vacuo.
The residue was
dissolved in ethyl acetate, washed with phosphate buffer (pH 7) and filtered.
After phase separation,
the organic phase was dried over anhydrous sodium sulfate, filtered and
concentrated under reduced
pressure. The crude product was purified by column chromatography (silica gel,
eluent: cyclohexane
/ ethyl acetate mixtures). The residue was suspended in acetonitrile, filtered
and dried invacuo. Yield:
1.14 g (91% purity, 26% of theory).
LC-MS (method 4): R1= 1.58 min; MS (ESIpos): m/z = 288 [M+H1+
Example 3.5D
(9aR,11aR)-6-Chloro-9a,10,11,11a-tetrahydro-2H-cyc10but44,5] [3] benzoxocino
[2, 1 -c] pyridin-
3(91-1)-one (single stereoisomer)
+NH
0
=
C
I
Enantiomer separation of 1.13 g of cis-6-chloro-9a,10,11,11a-tetrahydro-2H-
cyc10but44,51[3]benz-
oxocino[2,1-clpyridin-3(911)-one (racemate), Example 3.5C gave
single stereoisomer 1 (chiral SFC: Rt = 2.08 min, 85% ee): 404 mg,
single stereoisomer 2 (the title compound Example 3.5D) (chiral SFC: Rt = 2.23
min, 93% ee): 406
mg.
Separation method: SFC: column: Daicel Chiralpak OZ-H 5 um, 250 mm x 30 mm;
eluent: 60%
carbon dioxide /40% methanol; temperature: 40 C; flow rate: 100 ml/min; UV
detection: 210 nm.
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Analysis method: SFC: column: Daicel Chiralpak OZ-H 3 um, 100 mm x 4.6 mm;
eluent: 70%
carbon dioxide / 30% methanol; temperature: 40 C; flow rate: 3 ml/min; UV
detection: 210 nm.
LC-MS (method 4): R1= 1.59 min; MS (ESIpos): m/z = 288 [M+H1+
Example 3.6A1
5 -{ (2R)-2 -[(2 -B romo -4-chlorophenyl)methyl] -3,3,3 -trifluoropropoxy} -4 -
iodo -2 -methoxypyridine
(single stereoisomer)
CI F F
0
"
Br 1LLCH3
i
(Tributylphosphoranylidene)acetonitrile (3.2 g, 13.3 mmol, 1.5 eq.) and (2R)-2-
(2-bromo-4-
chlorobenzy1)-3,3,3-trifluoropropan-1-ol (single stereoisomer) (3.0 g, 8.8
mmol) were added at RT
to a solution of 4-iodo-6-methoxypyridin-3-ol (2.2 g, 8.8 mmol, 1.0 eq.) in
toluene (15 m1). The
resulting mixture was irradiated at 160 C for 2 h in a microwave reactor and
then cooled to RT. The
above reaction operation was repeated analogously for ten further batches. The
eleven batches,
altogether 33.0 g (97.3 mmol) of (2R)-2-(2-bromo-4-chlorobenzy1)-3,3,3-
trifluoropropan-1-ol)
(single stereoisomer), were combined, diluted with ethyl acetate and washed
with water and brine.
The organic layer was dried over anhydrous sodium sulfate, filtered and
concentrated under reduced
pressure. The residue was purified by column chromatography (silica gel,
eluent: petroleum ether /
ethyl acetate 9:1). Yield: 23.4 g (43% of theory).
1H-NMR (300 MHz, CDC13): 6 [ppm] = 7.60 (d, 1H), 7.50 (s, 1H), 7.30-7.17 (m,
3H), 4.18 (dd, 1H),
3.94-3.86 (m, 1H), 3.84 (s, 3H), 3.37-3.23 (m, 2H), 3.07-2.93 (m, 1H).
19F-NMR (282 MHz, CDC13): 6 [ppm] = -68.71 (s, 3F).
Example 3.6A2
5 -[(2R)-2 -(4-Chlorobenzy1)-3 ,3 ,3 -trifluoropropoxy] -4 -iodo-2-
methoxypyridine (single
stereoisomer)
CI F F
0
I "
C H3
i
Under argon atmosphere, a mixture of (2R)-2-(4-chlorobenzy1)-3,3,3-
trifluoropropan-1-ol (single
stereoisomer) (558 g, 2.34 mol) and 4-iodo-6-methoxypyridin-3-ol (1.76 kg,
7.01 mol, 3.0 eq.) in
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toluene (5.6 1) was treated dropwise (addition time: 1 h) under reflux with
(tributylphosphoranylidene)acetonitrile (847 g, 3.51 mol, 1.5 eq.). The
reaction mixture was stirred
for 2 h under reflux, cooled to 0 C and then methanol (366 ml) was added. The
reaction mixture was
diluted with toluene to a volume of 14.2 1 and then washed with 2 N aqueous
sodium hydroxide
solution (3 x 5.3 1). The organic phase was washed with 10% aqueous sodium
chloride solution (7.11)
and then concentrated under reduced pressure. The crude material was divided
into two equal
portions which were purified by column chromatography (12.0 kg silica gel,
eluent: dichloromethane
/ petroleum ether 3:2, then dichloromethane). Yield: 811 g (74% of theory).
Recovery of excess 4-
iodo-6-methoxypyridin-3-ol: The combined basic water phases were washed with
toluene (7.0 1).
The water phase was neutralized by addition of concentrated aqueous hydrogen
chloride solution and
the precipitate was then collected by filtration under reduced pressure. The
filter cake was washed
with water and then dried over phosphorus pentoxide under reduced pressure.
The crude material
was further purified by column chromatography (silica gel, eluent:
dichloromethane / acetone 4:1)
to yield pure 4-iodo-6-methoxypyridin-3-ol.
LC-MS (method 4): Rt = 2.64 min; MS (ESIpos): m/z = 471 [M+I-11+
1H-NMR (600 MHz, DM50-c/6): 6 [ppm] = 7.70 (s, 1H), 7.39-7.31 (m, 5H), 4.22
(dd, 1H), 3.89 (dd,
1H), 3.77 (s, 3H), 3.19 (dt, 1H), 3.14-3.04 (m, 2H).
Example 3.6B
(7 R) - 11 -Chloro-2-methoxy-7 -(trifluoromethyl)-7,8 -dihydro -6H-
[3]benzoxocino [2,1-c] pyridine
(single stereoisomer)
0
N
CH=
0' s'
441
CI
Bis(pinacolato)diboron (4.6 g, 18.2 mmol, 2.0 eq.), palladium(II) acetate
(0.20 g, 0.9 mmol, 0.1 eq.),
tricyclohexylphosphine (0.38 g, 1.4 mmol, 0.15 eq.) and potassium acetate
(2.67 g, 27.2 mmol, 3.0
eq.) were added to a solution of 5-{ [(2R)-2-(2-bromo-4-chlorobenzy1)-3,3,3-
trifluoropropylloxyl -4-
iodo-2-methoxypyridine (single stereoisomer) (5.0 g, 9.1 mmol) in NN-
dimethylformamide (100
m1). The resulting mixture was purged with nitrogen for 5 min and stirred at
80 C for 15 h under
nitrogen atmosphere. After cooling to RT, the reaction mixture was diluted
with ethyl acetate and
filtered through Celite . The filtrate was washed with water and brine, dried
over anhydrous sodium
sulfate, filtered and concentrated under reduced pressure. The residue was
purified by column
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chromatography (silica gel, eluent: petroleum ether / ethyl acetate 19:1).
Yield: 0.86 g (28% of
theory).
LC-MS (method 7): R1= 1.79 min; MS (ESIpos): m/z = 346 [M+I-11+
1H-NMR (400 MHz, CDC13): 6 [ppm] = 8.08 (s, 1H), 7.42-7.39 (m, 1H), 7.33-7.24
(m, 2H), 6.69 (s,
1H), 4.90-4.75 (m, 1H), 3.98 (s, 3H), 3.96-3.89 (m, 1H), 3.06 (d, 1H), 2.88-
2.63 (m, 1H), 2.60-2.51
(m, 1H).
19F-NMR (376 MHz, CDC13): 6 [ppm] = -70.92 (s, 3F).
Alternative synthetic route:
Two experiments of the same scale were run in parallel; the purification was
carried out with the
combined reaction mixtures.
Under argon atmosphere, a 10 1 round bottom flask was charged with potassium
pivalate (44.6 g,
318 mmol, 1.5 eq.) in N,N-dimethylacetamide (600 ml) and then the flask was
evacuated and back-
filled with argon three times. The mixture was heated to 150 C (oil bath
temperature: 180 C). A
second 5 1 round bottom flask was charged with 5-[(2R)-2-(4-chlorobenzy1)-
3,3,3-trifluoropropoxyl-
4-iodo-2-methoxypyridine (single stereoisomer) (100 g, 212 mmol) in N,N-
dimethylacetamide
(3.2 1) and the flask was evacuated and back-filled with argon three times.
Tetrakis(triphenylphosphine)palladium(0) (24.5 g, 21.2 mmol, 0.10 eq.) was
added and then the
mixture was added via pump addition (flow rate: 60 ml/min) to the preheated
potassium pivalate in
N,N-dimethylacetamide (throughout the addition the temperature was always kept
above 140 C).
After the addition, the reaction mixture was stirred for another 3.5 h at 150
C. The reaction mixture
was diluted with tert-butylmethyl ether (20 1) and 10% aqueous sodium chloride
solution (20 1) was
added. After stirring, the two phases were filtered over diatomaceous earth.
After phase separation,
the organic phase was washed with 10% aqueous sodium chloride solution (12 1),
dried over sodium
sulfate, filtered and concentrated under reduced pressure. The combined crude
material from two
batches was dissolved in dichloromethane and purified by column chromatography
(8.0 kg silica gel,
eluent: petroleum ether / ethyl acetate 19:1). The obtained material (65.5 g)
was triturated with
methanol (350 ml), the solid was filtered und reduced pressure and washed with
cold methanol (2 x
50 m1). The combined mother liquors were concentrated under reduced pressure
and the residue was
triturated with cold methanol (100 m1). The solid was washed with cold
methanol (2 x 20 ml) and
dried. Yield for the combined two batches: 41.6 g (29% of theory). The
combined mother liquors
were concentrated under reduced pressure and the residue was triturated with
cold methanol
(100 m1). The solid was washed with cold methanol (2 x 20 ml) and dried under
air atmosphere.
Yield for the combined two batches: 2.5 g (94% purity, 2% of theory). Combined
overall yield for
the two batches: 44.1 g (31% of theory).
LC-MS (method 18): Rt = 3.96 min; MS (ESIpos): m/z = 344 [M+I-11+
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1H-NMR (600 MHz, DMSO-d6): 6 [ppm] = 8.15 (s, 1H), 7.58-7.30 (m, 3H), 6.81 (s,
1H), 4.68 (br s,
1H), 4.22-4.03 /4.02-3.91 (2m, 1H), 3.88 (s, 3H), 3.25-3.15 /3.11-2.82 (2m,
2H), 2.74-2.57 / 2.48-
2.34 (2m, 1H, partially concealed). Additional signals of minor rotamers were
also detected.
Example 3.6C
(7 R) - 11 -Chloro-7-(trifluoromethyl)-7,8 -dihydro-3H-[3]benzoxocino [2, 1-c]
pyridin-2 (6H)-one
(single stereoisomer)
0
NH
0
=
CI
4-Toluenesulfonic acid monohydrate (0.75 g, 3.9 mmol, 1.8 eq.) and lithium
iodide (1.46 g,
10.9 mmol, 5.0 eq.) were added at RT to a solution of (7R)-11-chloro-2-methoxy-
7-
(trifluoromethyl)-7,8-dihydro-6H-[3]benzoxocino[2,1-c]pyridine (single
stereoisomer) (0.75 g, 2.2
mmol) in 1-butanol (40 m1). After stirring at 85 C for 15 h, the reaction
mixture was concentrated
under reduced pressure. The residue was dissolved in ethyl acetate and
saturated aqueous solution of
sodium carbonate and stirred for 10 min. The organic layer was washed with
water and brine, dried
over anhydrous sodium sulfate, filtered and concentrated under reduced
pressure. The residue was
purified by column chromatography (silica gel, eluent: petroleum ether / ethyl
acetate / methanol
5:19:1) to give 0.44 g of the crude product, which was triturated with
methanol and stirred for 30
min. The solid was collected by filtration, washed with methanol and dried
under air atmosphere.
Yield: 0.35 g (48% of theory).
LC-MS (method 9): R1= 1.34 min; MS (ESIpos): m/z = 330 [M+H1+
1H-NMR (400 MHz, DMSO-d6): 6 [ppm] = 11.55 (s, 1H), 7.54-7.25 (m, 4H), 6.35
(s, 1H), 4.75-4.55
(m, 1H), 4.11-3.78 (m, 1H), 3.12-2.86 (m, 2H), 2.69-2.55 (m, 1H).
19F-NMR (376 MHz, DMSO-d6): 6 [ppm] = -69.67 (s, 3F).
Alternative synthetic route:
Under argon atmosphere, 4-toluenesulfonic acid monohydrate (97.3 g, 511 mmol,
1.85 eq.) and
lithium chloride (58.6 g, 42.4 mmol, 5.0 eq.) were added at RT to a solution
of (7R)-11-chloro-2-
methoxy-7-(trifluoromethyl)-7,8 -dihydro -6H-[3]benzoxocino [2,1 -c] pyridine
(single stereoisomer)
(95.0 g, 276 mmol) in 2-propanol (1.2 1). After stirring under reflux for 16
h, the reaction mixture
was cooled to RT and then concentrated under reduced pressure to a volume of
about 250 ml. Water
(2.0 1) was added to the mixture and the resulting solution was extracted with
ethyl acetate (1.5 1and
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500 m1). The precipitate between the two layers was filtered off under reduced
pressure, washed with
ethyl acetate (200 ml) and dried under air atmosphere (yield: 53.2 g). After
phase separation, the
combined organic phases were concentrated under reduced pressure to a
remaining volume of about
100 ml. The precipitate was filtered off under reduced pressure, washed with
ethyl acetate and dried
under air atmosphere (yield: 23.5 g). The combined filtrates were washed with
saturated aqueous
sodium bicarbonate solution (1.11) and the precipitate, formed between the two
layers, was filtered
off under reduced pressure, washed with ethyl acetate and dried under air
atmosphere (yield: 9.2 g).
Combined yield: 85.9 g (94% of theory).
LC-MS (method 3): Rt = 2.54 min; MS (ESIpos): m/z = 330 [M+H1+
1H-NMR (600 MHz, DM50-d6): 6 [ppm] = 11.46 (br s, 1H), 7.59 (br s, 1H), 7.50
(br d, 1H), 7.42-
7.26 (m, 2H), 6.33 (br s, 1H), 4.64 (br d, 1H), 4.09-3.97 / 3.84 (m / br t,
1H), 3.26-3.14 / 3.08-2.89
(m, 2H), 2.81-2.69 / 2.58-2.51 (2m, 1H, partially concealed). Additional
signals of minor rotamers
were also detected.
Example 3.7A1
5 -{ (2R)-2-[(2-Bromo-4-chloro-3-fluorophenyl)methyl] -3,3,3 -
trifluoropropoxy} -4-iodo-2-
methoxypyridine (single stereoisomer)
C I F F
0
Br I C H 3
General Method 2 was carried out four times with (2R)-2-[(2-bromo-4-chloro-3-
fluorophenyl)methy11-3,3,3-trifluoropropan-l-ol (single stereoisomer) (920 mg,
2.60 mmol, 1.0 eq.),
4-iodo -6 -methoxypyridin-3 -ol (850 mg, 3.39 mmol, 1.3
eq.) and
(tributylphosphoranylidene)acetonitrile (1.0 ml, 3.9 mmol, 1.5 eq.) in toluene
(15 ml) at 120 C
including the following variations of the procedure: The crude mixtures of the
four reactions were
combined, concentrated under reduced pressure and purified by column
chromatography (silica gel,
eluent: cyclohexane / ethyl acetate 100:0 to 75:25). Yield: 2.71 g (46% of
theory).
LC-MS (method 5): R1= 1.78 min; MS (ESIpos): m/z = 569 [M+H1+
Example 3.7A2
5 -[(2R)-2 -(4-Chloro-3 -fluorobenzy1)-3 ,3 ,3 -trifluoropropoxy] -4 -iodo -2 -
methoxypyridine (single
stereoisomer)
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F
C I F F
OrNLI
I CH3
General Method 2 was carried out six times at 120 C for 3.0 h: one time with
(2R)-24(2-bromo-4-
chloro-3-fluorophenyl)methyl] -3,3,3 -trifluoropropan-1 -ol (single
stereoisomer) (50 mg, 189 umol,
1.0 eq.), 4-iodo-6-methoxypyridin-3-ol (77 mg, 92% purity, 283 umol, 1.5 eq.)
and (tributyl-
phosphoranylidene)acetonitrile (74 IA, 283 umol, 1.5 eq.) in toluene (2.5 ml),
four times with (2R)-
24(2 -bromo-4-chloro-3 -fluorophenyl)methyll -3,3,3 -trifluoropropan-1 -ol
(single stereoisomer) (360
mg, 1.36 mmol, 1.0 eq.), 4-iodo-6-methoxypyridin-3-ol (408 mg, 92% purity,
1.50 mmol, 1.1 eq.)
and (tributyl-phosphoranylidene)acetonitrile (530
2.0 mmol, 1.5 eq.) in toluene (17 ml) and one
time with (2R)-2{(2-bromo-4-chloro-3-fluorophenyl)methyll -3,3,3 -
trifluoropropan-1 -ol (single
stereoisomer) (360 mg, 1.36 mmol, 1.0 eq.), 4-iodo-6-methoxy-pyridin-3-ol (376
mg, 1.49 mmol,
1.5 eq.) and (tributylphosphoranylidene)-acetonitrile (530 IA, 2.0 mmol, 1.5
eq.) in toluene (17 ml)
including the following variations of the procedure: The crude mixtures of the
six reactions were
combined, concentrated under reduced pressure and purified by column
chromatography (silica gel,
eluent: cyclohexane / ethyl acetate 100:0 to 80:20). Yield: 1.35 g (40% of
theory).
LC-MS (method 4): Rt = 2.65 min; MS (ESIpos): m/z = 490 [M+F11+
Example 3.7B
(7 R) - 11 -Chloro-12-fluoro -2-methoxy-7-(trifluoromethyl)-7,8 -dihydro-6H-
[3] benzoxocino [2,1 -
c] pyridine (single stereoisomer)
0
N
H3
OC
F
C I
General Method 3a was carried with 5-{(2R)-24(2-bromo-4-chloro-3-
fluorophenyl)methyll -3,3,3-
trifluoropropoxy} -4-iodo-2-methoxypyridine (single stereoisomer) (2.71 g,
4.77 mmol, 1.0 eq.),
bis(pinacolato)diboron (1.94 g, 7.63 mmol, 1.6 eq.), palladium(II) acetate
(107 mg, 477 umol, 0.1
eq.), potassium acetate (1.40 g, 14.3 mmol, 3.0 eq.) in N,N-dimethylformamide
(70 ml) including the
following variations of the procedure: After stirring of the reaction mixture
at 80 C for 3 days, water
was added and the aqueous phase was extracted with ethyl acetate. The combined
organic phases
were dried over anhydrous sodium sulfate, filtered and concentrated under
reduced pressure. The
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crude mixture was purified by column chromatography (silica gel, eluent:
cyclohexane / ethyl acetate
95:0 to 20:80). Yield: 342 mg (99% purity, 20% of theory) and 1.33g (64%
purity, 49% of theory).
LC-MS (method 15): R1 = 2.03 min; MS (ESIpos): m/z = 362 [M+F11+
Alternative synthetic route:
5 -{(2R)-2 -(4-Chloro-3 -fluorobenzy1)-3 ,3 ,3 -trifluoropropoxy] -4 -iodo -2 -
methoxypyridine (single
stereoisomer) (1.25 g, 2.50 mmol, 1.0 eq.) and potassium pivalate (1.40 g,
10.0 mmol, 4.0 eq.) were
dissolved in N,N-dimethylacetamide (100 ml) and argon was passed through the
mixture for 10 min.
Subsequently, tetrakis(triphenylphosphine)palladium(0) (289 mg, 250 [tmol, 0.1
eq.) was added and
the mixture was stirred at 150 C for 2.5 h. The mixture was concentrated under
reduced pressure,
the residue was diluted with water and extracted with ethyl acetate. The
combined organic phases
were washed with brine, dried over anhydrous sodium sulfate, filtered and
concentrated under
reduced pressure. The crude product was purified by column chromatography
(silica gel, eluent:
cyclohexane / ethyl acetate 95:5 to 60:40). Yield: 420 mg (43% of theory).
LC-MS (method 5): R1= 1.55 min; MS (ESIpos): m/z = 362 [M+F11+
Example 3.7C
(7 R) - 11 -Chloro-12-fluoro -7-(trifluoromethyl)-7, 8-dihydro-3H-
Plbenzoxocino [2,1-c] pyridin-
2(611)-one (single stereoisomer)
0
NH
0
F
CI
General Method 4 was carried out with (7R)-11-chloro-12-fluoro-2-methoxy-7-
(trifluoromethyl)-
7,8-dihydro-6H-Mbenzoxocino[2,1-c]pyridine (single stereoisomer) (1.62 g, 71%
purity, 3.18
mmol) and sodium iodide (954 mg, 6.37 mmol, 2.0 eq.) in acetic acid (70 ml) at
100 C overnight
including the following variations of the procedure: Additional amounts of
sodium iodide (954 mg,
6.37 mmol, 2.0 eq.) were added to the reaction mixture and stirring was
continued at 100 C for 4 h.
The crude mixture was purified by column chromatography (silica gel, eluent:
dichloromethane /
methanol 98:2 to 80:20), followed by additional purification by column
chromatography (silica gel,
eluent: dichloromethane / methanol 98:2 to 80:20). Yield: 1.11 g (76% purity,
76% of theory).
LC-MS (method 5): R1= 1.15 min; MS (ESIpos): m/z = 348 [M+I-11+
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Example 3.8A
-{(2R)-2-(3 ,4 -Dichlorobenzy1)-3 ,3 ,3 -trifluoropropoxy] -4 -iodo -2 -
methoxypyridine (single
stereoisomer)
CI Ff F
CI
CH3
5 General Method 2 was carried out four times at 120 C for 6.0 h with (2R)-
2-(3,4-dichlorobenzy1)-
3,3,3-trifluoropropan-1-ol (single stereoisomer) (420 mg, 62% purity, 954
umol, 1.0 eq.), 4-iodo-6-
methoxypyridin-3-ol (390 mg, 92% purity, 1.43 mmol, 1.5 eq.) and (tributyl-
phosphoranylidene)-
acetonitrile (500 jil, 1.9 mmol, 2.0 eq.) in toluene (10 ml) including the
following variations of the
procedure: The crude mixtures of the four reactions were combined,
concentrated under reduced
pressure and purified twice by column chromatography (silica gel, eluent first
column: cyclohexane /
ethyl acetate 100:0 to 85:15, eluent second column: pure dichloromethane).
Yield: 0.48 g (25% of
theory).
LC-MS (method 4): Rt = 2.72 min; MS (ESIpos): m/z = 506 [M+1-11+
Example 3.8B
(7 R) - 11,12-Dichloro-2-methoxy-7 -(trifluoromethyl)-7, 8-dihydro-6H- [3]
benzoxocino [2,1-
c] pyridine (single stereoisomer)
0
N
O/C H3
CI
CI
5 -{(2R)-2-(3 ,4 -Dichlorobenzy1)-3 ,3 ,3 -trifluoropropoxy] -4 -iodo -2 -
methoxypyridine (single
stereoisomer) (378 mg, 747 umol, 1.0 eq.) and potassium pivalate (1.05 g, 7.47
mmol, 10.0 eq.) were
dissolved in N,N-dimethylacetamide (25 ml) and argon was passed through the
mixture for 10 min.
Subsequently, tetrakis(triphenylphosphine)palladium(0) (86.3 mg, 74.7 umol,
0.1 eq.) was added and
the mixture was stirred at 120 C overnight. The mixture was concentrated under
reduced pressure,
diluted with water and extracted with ethyl acetate. The combined organic
phases were dried over
anhydrous sodium sulfate, filtered and concentrated under reduced pressure.
The crude product was
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purified by column chromatography (silica gel, eluent: cyclohexane / ethyl
acetate 95:5 to 60:40).
Yield: 420 mg (47% purity, 16% of theory).
LC-MS (method 3): Rt = 4.22 min; MS (ESIpos): m/z = 378 [M+F11+
Example 3.8C
(7 R) - 11,12-Dichloro-7-(trifluoromethyl)-7, 8-dihydro -3H- [3] benzoxocino
[2,1 -c]pyridin-2 (6H)-one
(single stereoisomer)
0
NH
0
CI
CI
General Method 4 was carried out with (7R)-11,12-dichloro-2-methoxy-7-
(trifluoromethyl)-7,8-
dihydro-6H-[3]benzoxocino[2,1-clpyridine (single stereoisomer) (95.0 mg, 47%
purity, 118 umol,
1.0 eq.) and sodium iodide (88.5 mg, 590 umol, 5.0 eq.) in acetic acid (4.0
ml) at 100 C for 2.0 h
including the following variations of the procedure: The crude mixture was
purified by preparative
HPLC (reversed phase, eluent: water with 0.05% formic acid! acetonitrile 90:10
to 5:95). Yield: 21.0
mg (49% of theory).
LC-MS (method 3): Rt = 2.73; MS (ESIpos): m/z = 364 [M+H1+
1H-NMR (600 MHz, DM50-c/6): 6 [ppm] = 7.75-7.69 (m, 2H), 7.41 (d, 1H), 6.41
(s, 1H), 4.62 (dd,
1H), 3.82 (t, 1H), 3.07 (d, 2H), 3.02-2.86 (m, 1H).
Example 3.9A
5 4244 -Chlorobenzy1)-3 ,3 -difluoropropoxy] -4-iodo-2-methoxypyridine
(racemate)
CI F F
0
I
CH3
CY
Under argon atmosphere, a mixture of 2-(4-chlorobenzy1)-3,3-difluoropropan-1-
ol (racemate)
(3.50 g, 15.9 mmol) and 4-iodo-6-methoxypyridin-3-ol (3.98 g, 15.9 mmol, 1.0
eq.) in toluene
(50 ml) was treated with (tributylphosphoranylidene)acetonitrile (1.50 g, 23.8
mmol, 1.5 eq.). The
reaction mixture was stirred for 16 h under reflux, cooled to RT and purified
by column
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chromatography (silica gel, eluent: cyclohexane / ethyl acetate 100:0 to 95:5
to 9:1). Yield: 6.74 g
(94% of theory).
LC-MS (method 3): Rt = 4.48 min; MS (ESIpos): m/z = 454 [M+1-11+
Example 3.9B
11 -Chloro-7-(difluoromethyl)-2-methoxy-7,8 -dihydro -6H- [3] benzoxocino [2,1
-c] pyridine
(racemate)
0
N
=
CI
Under argon atmosphere, a round bottom flask was charged with cesium pivalate
(3.48 g, 14.9 mmol,
1.5 eq.) in N,N-dimethylacetamide (150 ml) and then the flask was frozen,
evacuated and back-filled
with argon three times. The mixture was then heated to 150 C. A second round
bottom flask was
charged with 542-(4-chlorobenzy1)-3,3-difluoropropoxy]-4-iodo-2-
methoxypyridine (racemate)
(4.5 g, 9.91 mmol) in N,N-dimethylacetamide (30 ml) and the flask was frozen,
evacuated and back-
filled with argon three times. Tetrakis(triphenylphosphine)palladium(0) (1.15
g, 0.99 mmol,
0.10 eq.) was added and the mixture was added via pump addition to the
preheated potassium pivalate
solution at 150 C within 1.0 h. After the addition, the reaction mixture was
stirred for another 5.5 h
at 150 C. The reaction mixture was diluted with saturated aqueous ammonium
chloride solution and
extracted with tert-butylmethyl ether. The combined organic layers were washed
with brine, dried
over anhydrous sodium sulfate, filtered and concentrated under reduced
pressure. The residue was
purified by column chromatography (silica gel, eluent: cyclohexane /toluene
100:0 to 3:1 to 1:1 to
1:3 to 0:100). Yield: 600 mg (17% of theory).
LC-MS (method 3): Rt = 3.68 min; MS (ESIpos): m/z = 326 [M+F11+
Example 3.9C
11 -Chloro-7-(difluoromethyl)-7,8 -dihydro -31/431benzoxocino [2,1 -c] pyridin-
2 (6H)-one (racemate)
0
NH
0
CI
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Under argon atmosphere, 4-toluenesulfonic acid monohydrate (596 mg, 3.13 mmol,
1.85 eq.) and
lithium chloride (359 mg, 8.47 mmol, 5.0 eq.) were added at RT to a solution
of 11-chloro-7-
(difluoromethyl)-2-methoxy-7,8-dihydro-6H-[3]benzoxocinop,1-c]pyridine
(racemate) (600 mg,
1.69 mmol) in 2-propanol (7.1 m1). After stirring under reflux for 16 h, the
reaction mixture was
cooled to RT and the resulting precipitate was filtered, washed with water and
dried. Yield: 515 mg
(98% of theory).
LC-MS (method 3): Rt = 2.26 min; MS (ESIpos): m/z = 312 [M+H1+
Example 4.1A
2-(Cyclopropyloxy)ethyl trifluoromethanesulfonate
0
0=Sv'
F*F
2,6-Dimethylpyridine (171 1.47 mmol, 1.5 eq.) and trifluoromethanesulfonic
anhydride (249
1.47 mmol, 1.5 eq.) were added dropwise under argon atmosphere at 0 C to a
solution of 2-
(cyclopropyloxy)ethan-1 -ol (100 mg, 0.98 mmol) in dichloromethane (4 m1). The
reaction mixture
was stirred at 0 C for 45 min, diluted with tert-butyl methyl ether and
extracted with a mixture (3:1)
of saturated aqueous solution of sodium chloride and aqueous hydrochloric acid
(1N). The combined
organic phases were dried over anhydrous sodium sulfate, filtered and
concentrated under reduced
pressure. The crude product was used without further purification. Yield: 262
mg.
'H-NMR (600 MHz, DM50-d6): 6 [ppm] = 4.44-4.37 (m, 2H), 3.72-3.65 (m, 2H),
3.45-3.38 (m, 1H),
0.55-0.45 (m, 4H).
Example 4.2A
2-[(1-Methylcyclopropyl)oxy] ethan-l-ol
C H 3
0 )
0 H
Magnesium turnings (474 mg, 19.5 mmol, 6.5 eq.) were placed into a round-
bottom flask and heated,
followed by addition of diethyl ether (5 ml) and some iodine. A solution of
1,2-dibromoethane (1.1
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ml, 13 mmol) in diethyl ether (5 ml) was added dropwise under reflux. After
stirring for additional 5
min, a solution of 2-(2-bromoethyl)-2-methyl-1,3-dioxolane (585 mg, 3.00 mmol)
in diethyl ether (5
ml) was added dropwise. The reaction mixture was stirred under reflux
overnight, cooled to RT,
diluted with diethyl ether, quenched cautiously with saturated aqueous
solution of ammonium
chloride and diluted with ethyl acetate and water. After phase separation, the
aqueous phase was
extracted with dichloromethane. The combined organic phases were washed with
brine, dried over
anhydrous sodium sulfate, filtered and concentrated under reduced pressure.
The crude product was
used without further purification. Yield: 211 mg (61% of theory).
'H-NMR (500 MHz, CDC13): 6 [ppm] = 3.70-3.63 (m, 2H), 3.60-3.54 (m, 2H), 1.40
(s, 3H), 0.81-
0.75 (m, 2H), 0.45-0.40 (m, 2H).
Example 4.2B
2-[(1-Methylcyclopropyl)oxy] ethyl trifluoromethanesulfonate
C H 3
0 )IV
0
0 V
0=S'
F*F
2,6-Dimethylpyridine (317 2.73 mmol, 1.5 eq.) and trifluoromethanesulfonic
anhydride (461
2.73 mmol, 1.5 eq.) were added dropwise under argon atmosphere at 0 C to a
solution of 24(1-
methylcyclopropyl)oxylethan-1-ol (211 mg, 1.82 mmol) in dichloromethane (3
m1). The reaction
mixture was stirred at 0 C for 1.5 h, diluted with tert-butyl methyl ether and
extracted with a mixture
(3:1) of saturated aqueous solution of sodium chloride and aqueous
hydrochloric acid (1 N). The
combined organic phases were dried over anhydrous sodium sulfate, filtered and
concentrated under
reduced pressure. The crude product was used without further purification.
Yield: 509 mg.
'H-NMR (600 MHz, CDC13): 6 [ppm] = 4.58-4.53 (m, 2H), 3.80-3.75 (m, 2H), 1.39
(s, 3H), 0.84-
0.78 (m, 2H), 0.46-0.40 (m, 2H).
Example 4.3A
[(25)-5 ,5 -Dimethyloxolan-2-yll methyl trifluoromethanesulfonate (single
stereoisomer)
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H 3C
H6-C H 3
0
0
0 0
0=S'
FAF
2,6-Dimethylpyridine (161 1.38 mmol, 1.5 eq.) and trifluoromethanesulfonic
anhydride (234
1.38 mmol, 1.5 eq.) were added dropwise under argon atmosphere at 0 C to a
solution of [(25)-5,5-
dimethyloxolan-2-yllmethanol (single stereoisomer) (120 mg, 0.92 mmol) in
dichloromethane (5
m1). The reaction mixture was stirred at 0 C for 45 min, diluted with tert-
butyl methyl ether and
extracted with a mixture (3:1) of saturated aqueous solution of sodium
chloride and aqueous
hydrochloric acid (1 N). The combined organic phases were dried over anhydrous
sodium sulfate,
filtered and concentrated under reduced pressure. The crude product was used
without further
purification. Yield: 265 mg.
'H-NMR (400 MHz, CDC13): 6 [ppm] = 4.53-4.46 (m, 1H), 4.44-4.37 (m, 1H), 4.33-
4.24 (m, 1H),
2.21-2.10 (m, 1H), 1.92-1.76 (m, 3H), 1.28 (s, 3H), 1.25 (s, 3H).
Example 5.1A
tert-Butyl (11-chloro-7-methyl-2-oxo-2,6,7,8-tetrahydro-3H-[3]benzoxocino [2,1-
c] pyridin-3 -
ypacetate (racemate)
H 3C N.r0C H3
I CH00 3
3
C
i
Potassium carbonate (226 mg, 1.63 mmol, 1.5 eq.) was added to a solution of 11-
chloro-7-methy1-
7,8-dihydro-3H-[3]benzoxocinop,1-c]pyridin-2(6H)-one (racemate) (300 mg, 1.09
mmol) and tert-
butyl bromoacetate (193
255 mg, 1.31 mmol, 1.2 eq.) in /V,N-dimethylformamide (3.0 m1). The
reaction mixture was stirred at 50 C overnight and then concentrated under
reduced pressure. The
residue was purified by column chromatography (silica gel, eluent: 0-95% ethyl
acetate /
cyclohexane). Yield: 281 mg (66% of theory).
LC-MS (method 4): R1= 2.10 min; MS (ESIpos): m/z = 390 [M+H1+
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Example 5.1B
tert-Butyl R7 R) - 11-chloro-7-methyl-2-oxo-2,6,7,8-tetrahydro-3H-
[3]benzoxocino [2,1-c] pyridin-3 -
yll acetate (single stereoisomer)
H 3C 0 N,ThrOi<C H 3
0 CF-C' 3
Ci
Enantiomer separation of 830 mg of tert-butyl (11-chloro-7-methy1-2-oxo-
2,6,7,8-tetrahydro-3H-
[3]benzoxocino [2,1-c]pyridin-3 -yl)acetate (racemate), Example 5.1A gave
single stereoisomer 1 (the title compound Example 5.1B) (chiral SFC: R1= 0.74
min, >99% ee): 414
mg,
single stereoisomer 2 (chiral SFC: R1= 1.39 min, >99% ee): 413 mg.
Separation method: SFC: column: Daicel Chiralpak AS-H 5 lam, 250 mm x 20 mm;
eluent: 65%
carbon dioxide / 35% methanol; temperature: 40 C; flow rate: 80 ml/min; UV
detection: 210 nm.
Analysis method: SFC: column: Daicel Chiralpak AS 5 lam, 100 mm x 4.6 mm;
eluent: 70% carbon
dioxide / 30% methanol; temperature: 40 C; flow rate: 3 ml/min; UV detection:
210 nm.
LC-MS (method 4): Rt = 2.08 min; MS (ESIpos): m/z = 390 [M+H1+
1H-NMR (400 MHz, DM50-d6): 6 [ppm] = 7.74 (s, 1H), 7.47-7.38 (m, 2H), 7.33-
7.30 (m, 1H), 6.35
(s, 1H), 4.65-4.50 (m, 2H), 4.48-4.41 (m, 1H), 3.30-3.25 (m, 1H), 2.70 (d,
1H), 2.33-2.24 (m, 1H),
2.09-1.94 (m, 1H), 1.44 (s, 9H), 0.89 (d, 3H). Additional signals of minor
rotamers were also
detected.
Example 5.2A
tert-Butyl 2 -(11-chloro -7-methy1-2-oxo-2,6,7,8 -tetrahydro -3H-
[3]benzoxocino [2,1-c] pyridin-3 -y1)-
3-[(25)-tetrahydro-2H-pyran-2-yllpropanoate (mixture of stereoisomers)
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0
0
H 3C
cC(:1C H3
N
H
0 C H 3
CI
A solution of sodium bis(trimethylsilyl)amide (0.8 ml, 1 M in tetrahydrofuran,
0.8 mmol, 1.25 eq.)
was added under argon atmosphere at -78 C dropwise to a solution of tert-butyl
(11-chloro-7-methyl-
2-oxo -2,6,7, 8-tetrahydro-3H- [3] benzoxocino [2,1-c] pyridin-3 -yl)acetate
(racemate) (250 mg, 0.641
mmol) in tetrahydrofuran (9.3 m1). After stirring at -78 C for 20 min, (25)-
tetrahydro-2H-pyran-2-
ylmethyl trifluoromethanesulfonate (single stereoisomer) (205 mg, 0.802 mmol,
1.25 eq.) (prepared
according to WO 2017/005725, example 3.3D) was added. The resulting reaction
mixture was stirred
at -78 C for 30 min, then slowly warmed to RT and stirred for 1 h. Glacial
acetic acid (55 [11, 58 mg,
0.96 mmol, 1.5 eq.) was added and the reaction mixture was concentrated under
reduced pressure.
The crude product was purified by preparative HPLC (reversed phase, eluent: 10-
90% acetonitrile /
water with 0.05% formic acid). Yield: 144 mg (46% of theory).
LC-MS (method 1): R1= 1.27 min; MS (ESIpos): m/z = 488 1M+H1+
Example 5.2B
2-(11-Chloro -7 -methy1-2 -oxo-2,6,7, 8-tetrahydro-3H- [3] benzoxocino [2,1 -
c] pyridin-3 -y1)-34(25)-
tetrahydro-2H-pyran-2-yllpropanoic acid (mixture of stereoisomers)
Ce3
0
H 3C
N
0
0
CI
Trifluoroacetic acid (3.1 ml) was added at RT to a solution of tert-butyl 2-
(11-chloro-7-methy1-2-
oxo-2,6,7, 8-tetrahydro -3H- [3] benzoxocino [2,1-c] pyridin-3 -y1)-3 - [(25)-
tetrahydro-2H-pyran-2 -
yllpropanoate (mixture of stereoisomers) (140 mg, 0.287 mmol) in
dichloromethane (3 m1). The
reaction mixture was stirred for 45 min and then concentrated under reduced
pressure. The residue
was dissolved in toluene and concentrated under reduced pressure. The residue
was dissolved in a
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mixture of acetonitrile and water and lyophilized. The crude product was used
without further
purification. Yield: 152 mg.
LC-MS (method 1): R1= 1.01 min; MS (ESIpos): m/z = 432 [M+F11+
Example 5.3A
tert-Butyl 2 - [(7R)-11-chloro -7 -methy1-2-oxo -2,6,7, 8-tetrahydro-3H-[3]
benzoxocino [2,1-c] pyridin-
3-yll -3-[(25)-tetrahydro-2H-pyran-2-yllpropanoate (mixture of two
diastereomers)
0
0
H 3 C cLi:3=C H3
N
H 3
0 C H3
Ci
A solution of sodium bis(trimethylsilyl)amide (0.51 ml, 1 M in
tetrahydrofuran, 0.51 mmol, 1.25 eq.)
was added dropwise under argon atmosphere at -78 C to a solution of tert-butyl
[(7R)-11-chloro-7-
methy1-2-oxo-2,6,7,8-tetrahydro-3H-[3]benz0x0cin0 [2,1-c] pyridin-3 -yl]
acetate (single
stereoisomer) (158 mg, 0.406 mmol) in tetrahydrofuran (12 m1). After stirring
at -78 C for 20 min,
a solution of (25)-tetrahydro-2H-pyran-2-ylmethyl trifluoromethanesulfonate
(single stereoisomer)
(130 mg, 0.508 mmol, 1.25 eq.) (prepared according to WO 2017/005725, example
3.3D) in
tetrahydrofuran (4 ml) was added. The resulting reaction mixture was stirred
at -78 C for 30 min,
then slowly warmed to RT and stirred for 1 h. Glacial acetic acid (35 [11, 37
mg, 0.61 mmol, 1.5 eq.)
was added and the reaction mixture was concentrated under reduced pressure.
The crude product was
purified by column chromatography (silica gel, eluent: 0-100% ethyl acetate /
cyclohexane). Yield:
112 mg (87% purity, 49% of theory).
LC-MS (method 4): Rt = 2.47 min; MS (ESIpos): m/z = 488 [M+1-11+
Example 5.3B
2-[(7R)-11 -Chloro -7 -methy1-2-oxo-2,6,7,8 -tetrahydro-3H-[3] benzoxocino
[2,1-c] pyridin-3 -yl] -3 -
[(25)-tetrahydro-2H-pyran-2-yllpropanoic acid (mixture of two diastereomers)
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Ce3
0
H 3C
N
0
0
CI
Lithium hydroxide (109 mg, 4.58 mmol, 23 eq.) was added to a solution of tert-
butyl 2-[(7R)-11-
chloro-7-methy1-2 -oxo -2,6,7,8-tetrahydro-3H- [3] benzoxocino [2,1-c] pyridin-
3 -yll -3 - [(25)-
tetrahydro -2H-pyran-2-yllpropanoate (mixture of two diastereomers) (111 mg,
87% purity, 0.197
mmol) in tetrahydrofuran (4.6 ml) and water (2.3 m1). The resulting reaction
mixture was shaken at
40 C for 22 h, then acidified with aqueous hydrochloric acid (1 N) and diluted
with ethyl acetate.
After phase separation, the aqueous phase was extracted with ethyl acetate.
The combined organic
phases were washed with brine, dried over anhydrous sodium sulfate, filtered
and concentrated under
reduced pressure. The crude product was used without further purification.
Yield: 22 mg (85% purity,
22% of theory).
LC-MS (method 4): R1= 1.86 / 1.89 min; MS (ESIpos): m/z = 432 [M+1-11+
Example 5.4A
tert-Butyl 2 - [(7R)-11-chloro -7 -methy1-2-oxo -2,6,7, 8-tetrahydro-3H-
[3]benzoxocino [2,1-c] pyridin-
3 -yll -3 - [(2R)-1,4 -dioxan-2 -yll propanoate (mixture of two diastereomers)
0
0
0
H 3C NcCCC H 3
H 3
00 C H 3
C
i
A solution of sodium bis(trimethylsilyl)amide (0.80 ml, 1 M in
tetrahydrofuran, 0.80 mmol, 1.25 eq.)
was added dropwise under argon atmosphere at -78 C to a solution of tert-butyl
[(7R)-11-chloro-7-
methy1-2-oxo-2,6,7,8-tetrahydro-3H-[3]benzoxocino [2,1-c] pyridin-3 -yl]
acetate (single
stereoisomer) (250 mg, 0.641 mmol) in tetrahydrofuran (18 m1). After stirring
at -78 C for 20 min,
.. a solution of (25)-1,4-dioxan-2-ylmethyl trifluoromethanesulfonate (single
stereoisomer) (201 mg,
0.802 mmol, 1.25 eq.) (prepared according to WO 2017/005725, example 3.5A) in
tetrahydrofuran
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(6 ml) was added. The resulting reaction mixture was stirred at -78 C for 30
min, then slowly warmed
to RT and stirred for 1 h. Glacial acetic acid (55 jil, 58 mg, 0.96 mmol, 1.5
eq.) was added and the
reaction mixture was concentrated under reduced pressure. The crude product
was purified by
column chromatography (silica gel, eluent: 0-100% ethyl acetate /
cyclohexane). Yield: 150 mg
(47% of theory).
LC-MS (method 4): R1= 2.18 min; MS (ESIpos): m/z = 490 [M+F11+
Example 5.4B
2-[(7R)-11 -Chloro -7-methy1-2-oxo-2,6,7,8 -tetrahydro-3H-[3]benzoxocino [2,1-
c] pyridin-3 -yl] -3 -
[(2R)-1,4-dioxan-2-yllpropanoic acid (mixture of two diastereomers)
C)
0
0
H 3C
N
0
0
C
I
Lithium hydroxide (147 mg, 6.12 mmol, 20 eq.) was added to a solution of tert-
butyl 2-[(7R)-11-
chloro-7-methy1-2 -oxo -2,6,7,8-tetrahydro-3H- [3] benzoxocino [2,1-c] pyridin-
3 -yll -3 - [(2R)-1,4 -
dioxan-2-yllpropanoate (mixture of two diastereomers) (150 mg, 0.306 mmol) in
tetrahydrofuran
(6.2 ml) and water (3.1 m1). The resulting reaction mixture was shaken at 40 C
overnight, then
acidified with aqueous hydrochloric acid (1 N) and diluted with ethyl acetate.
After phase separation,
the aqueous phase was extracted with ethyl acetate. The combined organic
phases were washed with
brine, dried over anhydrous sodium sulfate, filtered and concentrated under
reduced pressure. The
product was dried in vacuo and used in the next step without further
purification. Yield: 105 mg
(78% of theory).
LC-MS (method 4): R1= 1.61 min; MS (ESIpos): m/z = 434 [M+1-11+
Example 5.5A
tert-Butyl 2- [(7R)-11-chloro-7-methy1-2-oxo-2,6,7,8 -tetrahydro-3H- [3]
benzoxocino [2,1-c] pyridin-
3-y1]-4-(cyclopropyloxy)butanoate (mixture of two diastereomers)
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t.)
0
H 3C H 3
N
H3
0 C H3
Ci
A solution of lithium bis(trimethylsilyl)amide (0.31 ml, 1 M in
tetrahydrofuran, 0.31 mmol, 1.2 eq.)
was added under argon atmosphere at -78 C to a solution of tert-butyl [(7R)-11-
chloro-7-methy1-2-
oxo-2,6,7,8-tetrahydro-3H-[3]benzoxocino [2,1 -c] pyridin-3 -yl] acetate
(single stereoisomer)
(101 mg, 0.26 mmol) in tetrahydrofuran (3 m1). After stirring at -78 C for 15
min, 2-
(cyclopropyloxy)ethyl trifluoromethanesulfonate (91 mg, 0.39 mmol, 1.5 eq.)
was added dropwise.
The resulting reaction mixture was stirred at -78 C for 15 min and then
allowed to warm to RT. The
reaction mixture was quenched with water. After addition of ethyl acetate and
phase separation, the
aqueous phase was extracted with ethyl acetate. The combined organic phases
were washed with
brine, dried over anhydrous sodium sulfate, filtered and concentrated under
reduced pressure. The
residue was purified by preparative HPLC (reversed phase, eluent: acetonitrile
/ water gradient).
Yield: 12 mg (10% of theory).
LC-MS (method 4): Rt = 2.41 min; MS (ESIpos): m/z = 474 [M+H1+
Example 5.5B
2-[(7R)-11 -Chloro -7 -methy1-2-oxo-2,6,7,8 -tetrahydro-3H-[3] benzoxocino
[2,1 -c] pyridin-3 -yl] -4 -
(cyclopropyloxy)butanoic acid (mixture of two diastereomers)
0
H 3C H
N
0
0
CI
Lithium hydroxide (3 mg, 0.12 mmol, 4.5 eq.) was added at RT to a solution of
tert-butyl 2-[(7R)-
11 -chloro-7-methy1-2 -oxo-2,6,7, 8-tetrahydro-3H- [3] benzoxocino [2, 1 -c]
pyridin-3 -yll -4-
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(cyclopropyloxy)butanoate (mixture of two diastereomers) (12 mg, 0.03 mmol) in
a mixture of
tetrahydrofuran and water (3:1, 1 m1). The reaction mixture was stirred at RT
overnight and then
acidified with aqueous hydrochloric acid (1 N). After addition of ethyl
acetate and phase separation,
the aqueous phase was extracted with ethyl acetate. The combined organic
phases were dried over
.. anhydrous sodium sulfate, filtered and concentrated under reduced pressure.
The crude product was
used without further purification. Yield: 9 mg (82% of theory).
LC-MS (method 1): R1= 1.01 min; MS (ESIpos): m/z = 418 [M+F11+
Example 5.6A
tert-Butyl 2- [(7R)-11 -chloro -7-methy1-2-oxo -2,6,7,8 -tetrahydro -3H- [3]
benzoxocino [2,1 -c] pyridin-
3-y1]-4-(cyclobutyloxy)butanoate (mixture of two diastereomers)
0
0
H 3 c N 0 c H 3
hCH3
00 CH3
Ci
A solution of lithium bis(trimethylsilyl)amide (0.31 ml, 1 M in
tetrahydrofuran, 0.31 mmol, 1.2 eq.)
was added under argon atmosphere at -78 C to a solution of tert-butyl [(7R)-11-
chloro-7-methy1-2-
oxo-2,6,7,8-tetrahydro-3H-[3]benzoxocino [2,1 -c] pyridin-3 -yl] acetate
(single stereoisomer)
(101 mg, 0.26 mmol) in tetrahydrofuran (3 m1). After stirring at -78 C for 15
min, 2-
(cyclobutyloxy)ethyl trifluoromethanesulfonate (124 mg, 78% assumed purity of
crude product, 0.39
mmol, 1.5 eq.) (prepared according to WO 2016/146606, example 2.2A) was added
dropwise. The
resulting reaction mixture was stirred at -78 C for 15 min, then allowed to
warm to RT and stirred at
RT for 3 h. The reaction mixture was quenched with water. After addition of
ethyl acetate and phase
separation, the aqueous phase was extracted with ethyl acetate. The combined
organic phases were
washed with brine, dried over anhydrous sodium sulfate, filtered and
concentrated under reduced
pressure. The residue was purified by preparative HPLC (reversed phase,
eluent: acetonitrile / water
gradient). Yield: 80 mg (63% of theory).
LC-MS (method 3): Rt = 4.46 min; MS (ESIpos): m/z = 488 [M+H1+
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Example 5.6B
2-[(7R)-11 -Chloro -7-methy1-2-oxo-2,6,7,8 -tetrahydro-3H-[3]benzoxocino [2,1-
c] pyridin-3 -yl] -4 -
(cyclobutyloxy)butanoic acid (mixture of two diastereomers)
0
0
H 3C 0 H
N
0
0
CI
Trifluoroacetic acid (341 [11, 4.43 mmol, 20 eq.) was added at 0 C to a
solution of tert-butyl 2-[(7R)-
11-chloro-7-methy1-2 -oxo-2,6,7,8-tetrahydro-3H- [3] benzoxocino [2, 1-c]
pyridin-3 -yll -4 -
(cyclobutyloxy)-butanoate (mixture of two diastereomers) (108 mg, 0.22 mmol)
in dichloromethane
(3 m1). The reaction mixture was stirred for 3.5 hand then concentrated under
reduced pressure. The
residue was dissolved in toluene and concentrated under reduced pressure. The
residue was dissolved
in dichloromethane and concentrated under reduced pressure. Yield: 106 mg (83%
purity, 92% of
theory).
LC-MS (method 4): R1= 1.96 min; MS (ESIpos): miz = 432 [M+F11+
Example 5.7A
tert-Butyl R7 R) - 11-chloro -2-oxo-7-(trifluoromethyl)-2,6,7,8-tetrahydro-3H-
[3] benzoxocino [2,1-
clpyridin-3-yllacetate (single stereoisomer)
0
NrCIC H3
H 0 C H3 3
0
=
CI
Potassium carbonate (365 mg, 2.64 mmol, 1.5 eq.) and tert-butyl bromoacetate
(318 [IL 420 mg,
2.11 mmol, 1.2 eq.) were added to a solution of (7R)-11-chloro-7-
(trifluoromethyl)-7,8-dihydro-3H-
[3]benzoxocino[2,1-clpyridin-2(6H)-one (single stereoisomer) (580 mg, 1.76
mmol) in /V,N-
dimethylformamide (8 m1). The reaction mixture was stirred at 100 C for 50
min, cooled to RT and
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concentrated under reduced pressure. The residue was purified by column
chromatography (silica
gel, eluent: cyclohexane / ethyl acetate gradient). Yield: 635 mg (81% of
theory).
LC-MS (method 1): R1= 1.15 min; MS (ESIpos): m/z = 444 [M+H1+
Example 5.8A
tert-Butyl 2- [(7R)-11-chloro-2-oxo -7 -(trifluoromethyl)-2,6,7,8 -tetrahydro-
3H-[3]benzoxocino [2,1-
c] pyridin-3 -yll -3-(25)-5,5 -dimethyloxolan-2-yl]propanoate (mixture of two
diastereomers)
H 3C
CH 3
0
0
N OCH 3
hCH3
N. 0 CH3
=
CI
A solution of lithium bis(trimethylsilyl)amide (0.27 ml, 1 M in
tetrahydrofuran, 0.27 mmol, 1.2 eq.)
was added under argon atmosphere at -78 C to a solution of tert-butyl [(7R)-11-
chloro-2-oxo-7-
(trifluoromethyl)-2,6,7,8-tetrahydro-3H431benzoxocino [2,1-c] pyridin-3 -yll
acetate (single
stereoisomer) (100 mg, 0.23 mmol) in tetrahydrofuran (3 m1). After stirring at
-78 C for 15 min,
11(25)-5,5-dimethyloxolan-2-yllmethyl trifluoromethanesulfonate (single
stereoisomer) (100 mg,
0.36 mmol, 1.6 eq.) was added dropwise. The reaction mixture was stirred at -
78 C for 45 min, then
allowed to warm to 0 C and stirred at 0 C for another 80 min. The reaction
mixture was quenched
with saturated aqueous solution of ammonium chloride. After addition of ethyl
acetate and phase
separation, the aqueous phase was extracted with ethyl acetate. The combined
organic phases were
washed with brine, dried over anhydrous sodium sulfate, filtered and
concentrated under reduced
pressure. The residue was purified by preparative HPLC (reversed phase,
eluent: acetonitrile / water
gradient). Yield: 65 mg (52% of theory).
LC-MS (method 4): Rt = 2.55 / 2.58 min; MS (ESIpos): m/z = 556 [M+H1+
Example 5.8B
2-11(7R)-11 -Chloro -2 -oxo -7 -(trifluoromethyl)-2,6,7,8-tetrahydro-
3H431benzoxocino 112, 1-clpyridin-
3 -yll -3-R25)-5,5 -dimethyloxolan-2-yl]propanoic acid (mixture of two
diastereomers)
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H 3C
H 3
0
OH
0
0
=
CI
Lithium hydroxide (10 mg, 0.42 mmol, 2.0 eq.) was added at RT to a solution of
tert-butyl 2-[(7R)-
11-chloro-2-oxo-7-(trifluoromethyl) -2,6,7, 8-tetrahydro-3H-[3]benzoxocino
[2,1-c] pyridin-3 -yl] -3 -
[(25)-5,5-dimethyloxolan-2-yllpropanoate (mixture of two diastereomers) (117
mg, 0.21 mmol) in a
mixture of tetrahydrofuran and water (3:1, 4 m1). The reaction mixture was
stirred at RT overnight
and then acidified with aqueous hydrochloric acid (1 N). After removing all
volatiles under reduced
pressure, the residue was diluted with water. The forming precipitate was
filtered, washed with water
and dried in vacuo . Yield: 98 mg (87% purity, 81% of theory).
LC-MS (method 3): Rt = 3.56 / 3.61 min; MS (ESIpos): m/z = 500 [M+1-11+
Example 5.9A
tert-Butyl (2) -2- [(7R)-11-chloro-2 -oxo -7 -(trifluoromethyl)-2,6,7,8-
tetrahydro-3H-
[3] benzoxocino [2,1-c] pyridin-3 -yl] -2,3 ,5 -trideoxy-4 -0-methyl-L -glyce
ro -pentonate (mixture of
two diastereomers)
0'CH3
CH3
0
N OCH3
II hCH3
00 CH3
CI
A solution of lithium bis(trimethylsilyl)amide (0.46 ml, 1 M in
tetrahydrofuran, 0.46 mmol, 1.2 eq.)
was added under argon atmosphere at -78 C to a solution of tert-butyl [(7R)-11-
chloro-2-oxo-7-
(trifluoromethyl)-2,6,7,8-tetrahydro-3H-[3]benzoxocino [2,1-c] pyridin-3 -yll
acetate (single
stereoisomer) (170 mg, 0.38 mmol) in tetrahydrofuran (5 m1). After stirring at
-78 C for 15 min,
(25)-2-methoxypropyl trifluoromethanesulfonate (single stereoisomer) (128 mg,
0.58 mmol, 1.5 eq.)
(prepared according to WO 2014/154794, example 28.1A) was added dropwise. The
reaction mixture
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was stirred at -78 C for 15 min, then allowed to warm to RT and stirred at RT
for 1 h. The reaction
mixture was quenched with saturated aqueous solution of ammonium chloride.
After addition of
ethyl acetate and phase separation, the aqueous phase was extracted with ethyl
acetate. The combined
organic phases were washed with brine, dried over anhydrous sodium sulfate,
filtered and
.. concentrated under reduced pressure. The crude product was used in the next
step without further
purification. Yield: 235 mg.
LC-MS (method 4): Rt = 2.36 min; MS (ESIpos): m/z = 516 [M+H1+
Example 5.9B
(2 )-2- [(7R)-11-Chloro -2 -oxo-7-(trifluoromethyl)-2,6,7,8-tetrahydro-3H- [3]
benzoxocino [2,1-
clpyridin-3-yll -2,3 ,5 -trideoxy-4-0-methyl-L -glycero-p entonic acid
(mixture of two diastereomers)
C H3
H 3
0
0 H
N
0
0
=
CI
Trifluoroacetic acid (0.60 ml, 7.74 mmol, 20.0 eq.) was added at RT to a
solution of tert-butyl (2)-
2-[(7R)-11-chloro-2-oxo-7-(trifluoromethyl)-2,6,7,8-tetrahydro-3H-
[3]benzoxocino [2,1-c] pyridin-
3-yll -2,3,5-trideoxy-4-0-methyl-L-glycero-pentonate (mixture of two
diastereomers) (235 mg, 85%
assumed purity of crude product, 0.39 mmol) in dichloromethane (5 m1). The
reaction mixture was
stirred at RT for 4.5 h, concentrated under reduced pressure and coevaporated
two times with
dichloromethane. The crude product was used without further purification.
Yield: 255 mg (86%
purity).
LC-MS (method 4): R1= 1.82 / 1.84 min; MS (ESIpos): m/z = 460 [M+I-11+
Example 5.10A
tert-Butyl 2 - [(7R)-11-chloro -2 -oxo -7-(trifluoromethyl)-2,6,7,8-tetrahydro-
3H-[3]benzoxocino [2,1-
c] pyridin-3 -yll -4-(difluoromethoxy)butanoate (mixture of two diastereomers)
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)<H
0 F
0 .r0,CH 3
N
hCH3
00 CH3
Ci
1,1,3,3-Tetramethylguanidine (149 1.19 mmol, 3.0 eq.) was added under argon
atmosphere at RT
to a solution of (7 R) - 11-chloro-7-(trifluoromethyl)-7,8-dihydro-3H-
[3]benzoxocinop,1-clpyridin-
2(611)-one (single stereoisomer) (132 mg, 0.40 mmol) in 2-propanol / acetone
(4:1, 1.75 m1). The
mixture was stirred at RT for 15 min, followed by addition of tert-butyl 2-
bromo-4-
(difluoromethoxy)butanoate (racemate) (157 mg, 80% purity, 0.44 mmol, 1.1 eq.)
and of further 2-
propanol / acetone (4.1, 1.75 m1). The reaction mixture was stirred at RT for
3 days, pooled with a
preceding, analogously performed test campaign using (7R)-11-chloro-7-
(trifluoromethyl)-7,8-
dihydro-3H-[3]benzoxocino[2,1-clpyridin-2(6H)-one (single stereoisomer) (28
mg, 0.09 mmol) and
concentrated under reduced pressure. The residue was purified by column
chromatography (silica
gel, eluent: cyclohexane / ethyl acetate gradient). Yield: 188 mg (73% of
theory for both reaction
campaigns).
LC-MS (method 4): Rt = 2.43 min; MS (ESIpos): m/z = 538 [M+H1+
Example 5.10B
2-[(7R)-11 -Chloro -2 -oxo -7 -(trifluoromethyl)-2,6,7,8-tetrahydro-3H-
[3]benzoxocino [2,1 -clpyridin-
3-y1]-4-(difluoromethoxy)butanoic acid (mixture of two diastereomers)
)<H
0 F
0 N.r0H
0
0
CI
Lithium hydroxide (17 mg, 0.70 mmol, 2.0 eq.) was added at RT to a solution of
tert-butyl 2-[(7R)-
11 -chloro-2-oxo-7-(trifluoromethyl) -2,6,7, 8-tetrahydro-3H-[3]benzoxocino
[2,1 -c]pyridin-3 -yl] -4-
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(difluoro-methoxy)butanoate (mixture of two diastereomers) (188 mg, 0.35 mmol)
in a mixture of
tetrahydrofuran and water (3:1, 7 m1). The reaction mixture was stirred at RT
overnight and then
acidified with aqueous hydrochloric acid (1 N). After removing all volatiles
under reduced pressure,
the residue was diluted with water. The forming precipitate was filtered,
washed with water and dried
in vacuo . Yield: 140 mg (83% of theory).
LC-MS (method 1): Rt = 0.96 min; MS (ESIpos): m/z = 482 [M+1-11+
Example 5.11A
tert-Butyl
(2) -2 -[(7R)-11 -chloro-2-oxo-7-(trifluoromethyl)-2,6,7, 8-tetrahydro-3H-
[3] benzoxocino [2,1 -c] pyridin-3 -yl] -2,3 ,5 -trideoxy-4 -0-
(difluoromethyl)-D-glyce ro-pentonate
(mixture of two diastereomers)
)<H
0 F
H 3
0 I 0 CH 3
N
H 0 3
C H 3
"CI
1,1,3,3-Tetramethylguanidine (205
1.63 mmol, 3.0 eq.) was added under argon atmosphere at RT
to a solution of (7 R) - 11-chloro-7-(trifluoromethyl)-7,8-dihydro-3H-
[3]benzoxocinop,1-clpyridin-
2(611)-one (single stereoisomer) (181 mg, 0.55 mmol) in 2-propanol / acetone
(4:1, 2.5 m1). The
mixture was stirred at RT for 15 min, followed by addition of tert-butyl (4R)-
2-bromo-4-
(difluoromethoxy)-pentanoate (mixture of two diastereomers) (182 mg, 0.60
mmol, 1.1 eq.) and of
further 2-propanol / acetone (4:1, 2.5 m1). The reaction mixture was stirred
at RT for 2 days and
concentrated under reduced pressure. The residue was purified by column
chromatography (silica
gel, eluent: cyclohexane / ethyl acetate gradient). Yield: 224 mg (75% of
theory).
LC-MS (method 1): R1= 1.23 min; MS (ESIpos): m/z = 552 [M+1-11+
Example 5.11B
(2 )-2- [(7R)-11 -Chloro-2 -oxo-7 -(trifluoromethyl)-2,6,7,8-tetrahydro-3H-
[3] benzoxocino [2,1 -
clpyridin-3-yll -2,3,5-trideoxy-4-0-(difluoromethyl)-D-glycero-pentonic acid
(mixture of two
diastereomers)
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)<H
0 F
H 3
0 0 H
N
0
0
=
CI
Lithium hydroxide (19 mg, 0.80 mmol, 2.0 eq.) was added at RT to a solution of
tert-butyl
[ (7 R) - 11 -chloro-2-oxo-7-(trifluoromethyl)-2,6,7, 8-tetrahydro-3H-
[3]benzoxocino [2,1 -c] pyridin-3 -
y1]-2,3,5-trideoxy-4-0-(difluoromethyl)-D-glycero-pentonate (mixture of two
diastereomers) (220
mg, 0.40 mmol) in a mixture of tetrahydrofuran and water (3:1, 5 m1). The
reaction mixture was
stirred at RT overnight and then acidified with aqueous hydrochloric acid (1
N). After removing all
volatiles under reduced pressure, the residue was diluted with water. The
forming precipitate was
filtered, washed with water and dried in vacuo . Yield: 176 mg (89% of
theory).
LC-MS (method 4): Rt = 2.04 min; MS (ESIpos): m/z = 496 [M+1-11+
Example 5.12A
tert-Butyl (2) -2 -[(7R)-11 -chloro-2-oxo-7-(trifluoromethyl)-
2,6,7, 8-tetrahydro-3H-
[3] benzoxocino [2,1 -c] pyridin-3 -yl] -2,3 ,5 -trideoxy-4 -0-
(difluoromethyl)-L -glycero-pentonate
(mixture of two diastereomers)
)<H
0 F
H 3
0 I 0 H 3
II I N
H3
0 C H3
Ci
1,1,3,3-Tetramethylguanidine (151 [11, 1.20 mmol, 3.0 eq.) was added under
argon atmosphere at RT
to a solution of (7 R) - 11-chloro-7-(trifluoromethyl)-7,8-dihydro-3H-
[3]benzoxocinop,1-clpyridin-
2(611)-one (single stereoisomer) (133 mg, 0.40 mmol) in 2-propanol / acetone
(4:1, 2.5 m1). The
mixture was stirred at RT for 15 min, followed by addition of tert-butyl (45)-
2-bromo-4-
(difluoromethoxy)-pentanoate (mixture of two diastereomers) (138 mg, 0.44
mmol, 1.1 eq.) and of
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further 2-propanol / acetone (4:1, 2.5 m1). The reaction mixture was stirred
at RT for 3 days and
concentrated under reduced pressure. The residue was purified by preparative
HPLC (reversed phase,
eluent: acetonitrile / water gradient). Yield: 128 mg (58% of theory).
LC-MS (method 1): R1= 1.23 min; MS (ESIpos): m/z = 552 [M+1-11+
Example 5.12B
(2 )-2- [(7R)-11-Chloro -2 -oxo-7 -(trifluoromethyl)-2,6,7,8-tetrahydro-3H-
[3] benzoxocino [2,1-
c] pyridin-3 -yll -2,3 ,5 -trideoxy-4-0-(difluoromethyl)-L -glycero-pentonic
acid (mixture of two
diastereomers)
)<H
0 F
H3
0 1 OH
N
0
0
CI
Lithium hydroxide (11 mg, 0.46 mmol, 2.0 eq.) was added at RT to a solution of
tert-butyl
[(7 R) - 11 -chloro-2-oxo-7-(trifluoromethyl)-2,6,7, 8-tetrahydro-3H-
[3]benzoxocino [2,1-c] pyridin-3 -
y11-2,3,5-trideoxy-4-0-(difluoromethyl)-L-glycero-pentonate (mixture of two
diastereomers) (128
mg, 0.23 mmol) in a mixture of tetrahydrofuran and water (3:1, 5 m1). The
reaction mixture was
stirred at RT overnight and then acidified with aqueous hydrochloric acid (1
N). After removing all
volatiles under reduced pressure, the residue was diluted with water. The
forming precipitate was
filtered, washed with water and dried in vacuo . Yield: 93 mg (89% purity, 72%
of theory).
LC-MS (method 4): R1= 1.95 min; MS (ESIpos): m/z = 496 [M+H1+
Example 5.13A
tert-Butyl 2- [(7R)-11-chloro-2-oxo -7 -(trifluoromethyl)-2,6,7,8 -tetrahydro-
3H-[3]benzoxocino [2,1-
clpyridin-3-y11-4-(2,2-difluoroethoxy)butanoate (mixture of two diastereomers)
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F.,. F
0
H 3
N
hC H3
0 C H 3
0
Ci
A solution of lithium bis(trimethylsilyl)amide (0.41 ml, 1 M in
tetrahydrofuran, 0.41 mmol, 1.2 eq.)
was added under argon atmosphere at -78 C to a solution of tert-butyl [(7R)-11-
chloro-2-oxo-7-
(trifluoromethyl)-2,6,7, 8-tetrahydro-3H-[3] benzoxocino [2,1-c] pyridin-3 -
yll acetate (single
stereoisomer) (150 mg, 0.34 mmol) in tetrahydrofuran (5 m1). After stirring at
-78 C for 15 min, 2-
(2,2-difluoroethoxy)ethyl trifluoromethanesulfonate (131 mg, 0.51 mmol, 1.5
eq.) (prepared
according to WO 2016/046164, example 24.1A) was added dropwise. The reaction
mixture was
stirred at -78 C for 30 min and then allowed to warm to 0 C. The reaction
mixture was quenched
with saturated aqueous solution of ammonium chloride. After addition of ethyl
acetate and phase
separation, the aqueous phase was extracted with ethyl acetate. The combined
organic phases were
washed with brine, dried over anhydrous sodium sulfate, filtered and
concentrated under reduced
pressure. The residue was purified by column chromatography (silica gel,
eluent: cyclohexane / ethyl
acetate gradient). Yield: 126 mg (66% of theory).
LC-MS (method 1): R1= 1.21 min; MS (ESIpos): m/z = 552 [M+1-11+
Example 5.13B
2-[(7R)-11-Chloro -2 -oxo -7 -(trifluoromethyl)-2,6,7,8-tetrahydro-
31/431benzoxocino [2,1-c] pyridin-
3-y1]-4-(2,2-difluoroethoxy)butanoic acid (mixture of two diastereomers)
F F
0
j.r0 H
N
0
0
=
CI
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Lithium hydroxide (11 mg, 0.44 mmol, 2.0 eq.) was added at RT to a solution of
tert-butyl 2-[(7R)-
11-chloro-2-oxo-7-(trifluoromethyl) -2,6,7, 8-tetrahydro-3H-[3]benzoxocino
[2,1-c] pyridin-3 -yl] -4 -
(2,2-difluoroethoxy)butanoate (mixture of two diastereomers) (125 mg, 0.22
mmol) in a mixture of
tetrahydrofuran and water (3:1, 5 m1). The reaction mixture was stirred at RT
overnight, then
acidified with aqueous hydrochloric acid (1 N) and diluted with water. After
removing all volatiles
under reduced pressure, the forming precipitate was filtered, washed with
water and dried in vacuo.
Yield: 85 mg (78% of theory).
LC-MS (method 4): R1= 1.85 min; MS (ESIpos): m/z = 496 [M+H1+
Example 5.14A
tert-Butyl 2- [(7R)-11-chloro-2-oxo -7 -(trifluoromethyl)-2,6,7,8 -tetrahydro-
3H-[3]benzoxocino [2,1-
c] pyridin-3 -y11-4- Rpropan-2-yl)oxy] butanoate (mixture of two
diastereomers)
C H 3
0 H 3
0
H 3
N
H3
C H3
0
=
Ci
A solution of lithium bis(trimethylsilyl)amide (0.46 ml, 1 M in
tetrahydrofuran, 0.46 mmol, 1.2 eq.)
was added under argon atmosphere at -78 C to a solution of tert-butyl [(7R)-11-
chloro-2-oxo-7-
(trifluoromethyl)-2,6,7,8-tetrahydro-3H-[3]benzoxocino [2,1-c] pyridin-3 -yll
acetate (single
stereoisomer) (170 mg, 0.38 mmol) in tetrahydrofuran (5 m1). After stirring at
-78 C for 15 min, 2-
Rpropan-2-yl)oxylethyl trifluoromethanesulfonate (136 mg, 0.58 mmol, 1.5 eq.)
(prepared according
to WO 2016/146606, example 2.1A) was added dropwise. The resulting reaction
mixture was stirred
at -78 C for 15 min, then allowed to warm to RT and stirred at RT for 1.5 h.
The reaction mixture
was quenched with saturated aqueous solution of ammonium chloride. After
addition of ethyl acetate
and phase separation, the aqueous phase was extracted with ethyl acetate. The
combined organic
phases were washed with brine, dried over anhydrous sodium sulfate, filtered
and concentrated under
reduced pressure. The crude product was used in the next step without further
purification. Yield:
260 mg.
LC-MS (method 1): R1= 1.29 min; MS (ESIpos): m/z = 530 [M+H] +.
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Example 5.14B
2-[(7R)-11-Chloro-2-oxo-7-(trifluoromethyl)-2,6,7,8-tetrahydro-3H-
[3]benzoxocinop,1-clpyridin-
3-yll-4-[(propan-2-ypoxylbutanoic acid (mixture of two diastereomers)
C H3
OLC H 3
0
0 H
N
0
0
CI
Lithium hydroxide (19 mg, 0.79 mmol, 2.0 eq.) was added at RT to a solution of
tert-butyl 2-[(7R)-
11-chloro-2-oxo-7-(trifluoromethyl) -2,6,7, 8-tetrahydro-3H-[3] benzoxocino
[2,1-c] pyridin-3 -yl] -4 -
Rpropan-2-yl)oxylbutanoate (mixture of two diastereomers) (260 mg, 80% assumed
purity of crude
product, 0.39 mmol) in a mixture of tetrahydrofuran and water (3:1, 8 m1). The
reaction mixture was
stirred at RT overnight, then acidified with aqueous hydrochloric acid (1 N)
and diluted with water.
After removing all volatiles under reduced pressure, the aqueous phase was
extracted with ethyl
acetate. The combined organic phases were washed with brine, dried over
anhydrous sodium sulfate,
filtered and concentrated under reduced pressure. The crude product was used
without further
purification. Yield: 166 mg (92% purity, 82% of theory).
LC-MS (method 4): R1= 1.97 min; MS (ESIpos): m/z = 474 [M+1-11+
Example 5.15A
tert-Butyl 4-tert-butoxy-2-[(7R)-11-chloro-2-oxo-7-(trifluoromethyl)-2,6,7,8-
tetrahydro-3H-
[3]benzoxocino[2,1-clpyridin-3-yllbutanoate (mixture of two diastereomers)
C H3
)<C H3
0 C H3
0
.rC)C H3
N
hC H3
0 C H3
0
"CI
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A solution of lithium bis(trimethylsilyl)amide (0.41 ml, 1 M in
tetrahydrofuran, 0.41 mmol, 1.2 eq.)
was added under argon atmosphere at -78 C to a solution of tert-butyl [(7R)-11-
chloro-2-oxo-7-
(trifluoromethyl)-2,6,7, 8-tetrahydro-3H-[3]benzoxocino [2,1-c] pyridin-3 -yll
acetate (single
stereoisomer) (150 mg, 0.34 mmol) in tetrahydrofuran (5 m1). After stirring at
-78 C for 15 min, 2-
tert-butoxyethyl trifluoromethanesulfonate (190 mg, 80% assumed purity of
crude product, 0.61
mmol, 1.8 eq.) (prepared according to WO 2016/146606, example 2.3A) was added
dropwise. The
reaction mixture was stirred at -78 C for 15 min, then allowed to warm to RT
and stirred at RT for
20 min. The reaction mixture was quenched with saturated aqueous solution of
ammonium chloride.
After addition of ethyl acetate and phase separation, the aqueous phase was
extracted with ethyl
acetate. The combined organic phases were washed with brine, dried over
anhydrous sodium sulfate,
filtered and concentrated under reduced pressure. The residue was dissolved in
dichloromethane and
filtered. The filtrate was purified by column chromatography (silica gel,
eluent: cyclohexane / ethyl
acetate gradient), the precipitate was purified by preparative HPLC (reversed
phase, eluent:
acetonitrile / water gradient), and both batches were combined. Yield: 81 mg
(44% of theory).
LC-MS (method 4): Rt = 2.56 min; MS (ESIpos): m/z = 544 [M+F11+
Example 5.15B
4-tert-Butoxy-2-{(7R)-11-chloro-2-oxo -7 -(trifluoromethyl)-2,6,7, 8-
tetrahydro-3H-
[31benzoxocino[2,1-clpyridin-3-yllbutanoic acid (mixture of two diastereomers)
C H 3
)c-C H 3
0 CH 3
0
H
N
0
0
CI
Lithium hydroxide (7 mg, 0.29 mmol, 2.0 eq.) was added at RT to a solution of
tert-butyl 4-tert-
butoxy-2-[(7 R)-11-chloro -2 -oxo -7 -(trifluoromethyl)-2,6,7,8-tetrahydro-3H-
[3] benzoxocino [2,1-
c] pyridin-3 -yllbutanoate (mixture of two diastereomers) (80 mg, 0.15 mmol)
in a mixture of
tetrahydrofuran and water (3:1, 4 m1). The reaction mixture was stirred at RT
overnight, then
acidified with aqueous hydrochloric acid (1 N) and diluted with water. After
removing all volatiles
under reduced pressure, the aqueous phase was cooled in an ice bath. The
forming precipitate was
filtered, washed with water and dried in vacuo . Yield: 182 mg (72% of
theory).
LC-MS (method 4): Rt = 2.06 min; MS (ESIpos): m/z = 488 [M+H1+
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Example 5.16A
tert-Butyl 2- [(7R)-11-chloro-2-oxo -7-(trifluoromethyl)-2,6,7,8 -tetrahydro-
3H431benzoxocino [2,1-
clpyridin-3-y11-4-(cyclopropyloxy)butanoate (mixture of two diastereomers)
0
0
N 0 H 3
H 0 3
0 C H 3
=
Ci
A solution of lithium bis(trimethylsilyl)amide (0.41 ml, 1 Mm tetrahydrofuran,
0.41 mmol, 1.2 eq.)
was added under argon atmosphere at -78 C to a solution of tert-butyl [(7R)-11-
chloro-2-oxo-7-
(trifluoromethyl)-2,6,7,8-tetrahydro-3H431benzoxocino [2,1-c] pyridin-3 -yll
acetate (single
stereoisomer) (150 mg, 0.34 mmol) in tetrahydrofuran (5 m1). After stirring at
-78 C for 15 min,
2-(cyclopropyloxy)ethyl trifluoromethanesulfonate (127 mg, 0.54 mmol, 1.6 eq.)
was added
dropwise. The reaction mixture was stirred at -78 C for 30 min and then
allowed to warm to RT. The
reaction mixture was quenched with saturated aqueous solution of ammonium
chloride. After
addition of ethyl acetate and phase separation, the aqueous phase was
extracted with ethyl acetate.
The combined organic phases were washed with brine, dried over anhydrous
sodium sulfate, filtered
and concentrated under reduced pressure. The residue was purified by column
chromatography
(silica gel, eluent: cyclohexane / ethyl acetate gradient). Yield: 94 mg (53%
of theory).
LC-MS (method 3): Rt = 4.34 min; MS (ESIpos): m/z = 528 [M+H1+
Example 5.16B
2-[(7R)-11 -Chloro -2 -oxo -7-(trifluoromethyl)-2,6,7,8-tetrahydro-
31/431benzoxocino [2,1-clpyridin-
3-y11-4-(cyclopropyloxy)butanoic acid (mixture of two diastereomers)
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0
0
H
N
0
0
CI
Lithium hydroxide (9 mg, 0.36 mmol, 2.0 eq.) was added at RT to a solution of
tert-butyl 2-[(7R)-
11-chloro-2-oxo-7-(trifluoromethyl) -2,6,7, 8-tetrahydro-3H-[3]benzoxocino
[2,1-c] pyridin-3 -yl] -4 -
(cyclopropyloxy)butanoate (mixture of two diastereomers) (94 mg, 0.18 mmol) in
a mixture of
tetrahydrofuran and water (3:1, 5 m1). The reaction mixture was stirred at RT
for 2 days, then
acidified with aqueous hydrochloric acid (1 N) and diluted with water. After
removing all volatiles
under reduced pressure, the forming precipitate was filtered, washed with
water and dried in vacuo.
Yield: 66 mg (79% of theory).
LC-MS (method 4): R1= 1.91 min; MS (ESIpos): m/z = 472 [M+1-11+
Example 5.17A
tert-Butyl 2- [(7R)-11-chloro-2-oxo -7 -(trifluoromethyl)-2,6,7,8 -tetrahydro-
3H-[3]benzoxocino [2,1-
c] pyridin-3 -yll -4- [(1-methylcyclopropyl)oxy] butanoate (mixture of two
diastereomers)
CH3
0)V
0
.rOCH 3
N
ICH3
0 CH3
0
CI
A solution of lithium bis(trimethylsilyl)amide (0.84 ml, 1 M in
tetrahydrofuran, 0.84 mmol, 1.2 eq.)
was added under argon atmosphere at -78 C to a solution of tert-butyl [(7R)-11-
chloro-2-oxo-7-
(trifluoromethyl)-2,6,7,8-tetrahydro-3H-[3]benzoxocino [2,1-c] pyridin-3 -yll
acetate (single
stereoisomer) (310 mg, 0.70 mmol) in tetrahydrofuran (10 m1). After stirring
at -78 C for 15 min, 2-
[(1-methylcyclopropyl)oxylethyl trifluoromethanesulfonate (260 mg, 1.05 mmol,
1.5 eq.) was added
dropwise. The reaction mixture was stirred at -78 C for 15 min, then allowed
to warm to RT and
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stirred at RT for 2 h. The reaction mixture was quenched with saturated
aqueous solution of
ammonium chloride. After addition of ethyl acetate and phase separation, the
aqueous phase was
extracted with ethyl acetate. The combined organic phases were washed with
brine, dried over
anhydrous sodium sulfate, filtered and concentrated under reduced pressure.
The residue was
combined with crude product from a preceding, analogously performed test
campaign using 30 mg
(0.07 mmol) of tert-butyl R7 R) - 11-chloro-2-oxo-7-(trifluoromethyl)-
2,6,7,8-tetrahydro-3H-
[3]benzoxocino[2,1-c]pyridin-3-yl]acetate (single stereoisomer) and purified
by column
chromatography (silica gel, eluent: cyclohexane / ethyl acetate gradient).
Yield: 260 mg (67% of
theory over both batches).
LC-MS (method 4): Rt = 2.51 min; MS (ESIpos): m/z = 542 [M+I-11+
Example 5.17B
2-[(7R)-11 -Chloro -2 -oxo -7-(trifluoromethyl)-2,6,7,8-tetrahydro-3H-
[3]benzoxocino [2,1-c] pyridin-
3-y1]-4-[(1-methylcyclopropyl)oxy]butanoic acid (mixture of two diastereomers)
C H3
O)V
0
H
N
0
0
CI
Lithium hydroxide (23 mg, 0.94 mmol, 2.0 eq.) was added at RT to a solution of
tert-butyl 2-[(7R)-
11-chloro-2-oxo-7-(trifluoromethyl) -2,6,7,8-tetrahydro-3H-[3]benzoxocino [2,1-
c] pyridin-3 -yl] -4 -
[(1-methylcyclopropyl)oxy]butanoate (mixture of two diastereomers) (260 mg,
0.47 mmol) in a
mixture of tetrahydrofuran and water (3:1, 5 m1). The reaction mixture was
stirred at RT for 26 h,
then acidified with aqueous hydrochloric acid (1 N) and diluted with water.
After removing all
volatiles under reduced pressure, the aqueous phase was extracted with ethyl
acetate. The combined
organic phases were washed with brine, dried over anhydrous sodium sulfate,
filtered and
concentrated under reduced pressure. The crude product was used without
further purification. Yield:
182 mg (93% purity, 74% of theory).
LC-MS (method 4): Rt = 2.02 min; MS (ESIpos): m/z = 486 [M+I-11+
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Example 5.18A
Ethyl 2- [(7R)-11 -chloro -2 -oxo -7 -(trifluoromethyl)-2,6,7, 8-
tetrahydro-3H431benzoxocino [2,1-
clpyridin-3-y11-4-(cyclobutyloxy)butanoate (mixture of two diastereomers)
oJ11:17
0 r() H3
N
0
0
CI
General Method 7 was carried out with (7R)-11-chloro-7-(trifluoromethyl)-7,8-
dihydro-3H-
[3]benzoxocino[2,1-clpyridin-2(6H)-one (single stereoisomer) (99.5 mg, 302
umol, 1.0 eq.), ethyl
2-bromo-4-(cyclobutyloxy)butanoate (racemate) (120 mg, 75% purity, 453 [Imo',
1.125 eq.) and
1,1,3,3-tetramethylguanidine (110 jil, 910 umol, 3.0 eq.) in a mixture of 2-
propanol / acetone (4:1,
3.75 ml) including the following variations of the procedure: After stirring
for 3 days, additional
amounts of 1,1,3,3-tetramethylguanidine (100 jil, 800 umol, 2.65 eq.) were
added and the mixture
was stirred overnight. The crude mixture was purified by preparative HPLC
(reversed phase, eluent:
water with 0.05% formic acid / acetonitrile 80:20 to 5:95). Yield: 85 mg (54%
of theory).
LC-MS (method 1): Rt = 1.22 min; MS (ESIpos): m/z = 514 [M+H1+
Example 5.18B
2-[(7R)-11 -Chloro -2 -oxo -7 -(trifluoromethyl)-2,6,7,8-tetrahydro-
31/431benzoxocino [2,1 -c] pyridin-
3-y1]-4-(cyclobutyloxy)butanoic acid (mixture of two diastereomers)
,17,7
0 H
N
0
0
CI
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General Method 10 was carried out with ethyl 2-[(7R)-11-chloro-2-oxo-7-
(trifluoromethyl)-2,6,7,8-
tetrahydro-3H-[3]benzoxocino 112, 1 -c] pyridin-3 -yl] -4 -
(cyclobutyloxy)butanoate (mixture of two
diastereomers) (85.0 mg, 165 umol, 1.0 eq.) and lithium hydroxide (19.8 mg,
827 umol, 5.0 eq.) in
a mixture of ethanol / tetrahydrofuran (2:1, 3 m1). Yield: 76.2 mg (95% of
theory).
LC-MS (method 1): R1= 1.06 min; MS (ESIpos): m/z = 486 [M+H1+
Example 5.19A
tert-Butyl 2 - [(7R)-11 -chloro -2 -oxo -7 -(trifluoromethyl)-2,6,7, 8-
tetrahydro-3H-[3] benzoxocino 112, 1 -
c] pyridin-3 -yll -4-[(1-methylcyclobutypoxylbutanoate (mixture of two
diastereomers)
\ '
CH 3
0
N OCH 3
hCH3
00 CH 3
=
Ci
General Method 9 was carried out with tert-butyl (7 R) - 11-chloro-2-oxo-7-
(trifluoromethyl)-2,6,7,8-
tetrahydro-3H-[3]benzoxocino [2, 1 -c] pyridin-3 -yl] acetate (single stere oi
some r) (150 mg, 338 umol),
2-[(1-methylcyclobutypoxylethyl trifluoromethanesulfonate (148 mg, 90% purity,
507 umol,
1.5 eq., preparation according to WO 2017/037051, example 3.10C) and lithium
bis(trimethylsilyl)amide (410 jil, 1.0 M, 410 umol, 1.2 eq.) in
tetrahydrofuran (6.1 ml) including the
following variations of the procedure: The reaction mixture was worked up by
addition of saturated
aqueous ammonium chloride solution, water and ethyl acetate, extracted with
ethyl acetate, dried
over anhydrous sodium sulfate, filtered and concentrated under reduced
pressure. The crude mixture
was purified by preparative HPLC (reversed phase, eluent: water with 0.05%
formic acid /
acetonitrile 80:20 to 5:95). Yield: 49.7 mg (27% of theory).
LC-MS (method 1): R1= 1.38 min; MS (ESIpos): m/z = 556 [M+F11+
Example 5.19B
2-11(7R)-11 -Chloro -2 -oxo -7 -(trifluoromethyl)-2,6,7,8-tetrahydro-3H-[3]
benzoxocino [2,1 -c] pyridin-
3-yll -4-[(1-methylcyclobutypoxylbutanoic acid (mixture of two diastereomers)
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CH 3
0
OH
N
0
0
CI
General Method 10 was carried out with tert-butyl 2-R7R)-11-chloro-2-oxo-7-
(trifluoromethyl)-
2,6,7,8-tetrahydro-3H-[3]benzoxocino [2,1 -c] pyridin-3 -yl] -4-R 1 -
methylcyclobutypoxy] butanoate
(mixture of two diastereomers) (49.5 mg, 89.0 umol, 1.0 eq.) and lithium
hydroxide (10.7 mg, 445
umol, 5.0 eq.) in a mixture of ethanol / tetrahydrofuran (2:1, 3 m1). Yield:
50.0 mg (quantitative of
theory).
LC-MS (method 1): R1= 1.10 min; MS (ESIpos): m/z = 500 [M+1-11+
Example 5.20A
Mixture of ethyl 4-0-[tert-butyl(dimethypsilyll -2-[(7R)-11-chloro-2-oxo-7-
(trifluoromethyl)-
2,6,7,8-tetrahydro -3H-[3] benzoxocino [2,1 -c] pyridin-3 -yl] -2,3 ,5 -tride
oxy-5 ,5 ,5 -trifluoropentonate
(mixture of stereoisomers) and isopropyl 4-0-[tert-butyl(dimethyl)silyll -2-
[(7R)-11-chloro-2-oxo-7-
(trifluoromethyl)-2,6,7,8-tetrahydro-3H-[3]benzoxocino [2,1 -c] pyridin-3 -yll
-2,3,5 -trideoxy-5,5,5 -
trifluoropentonate (mixture of stereoisomers)
F F3 cH3 F F3 cH3
Si CH3
0'Si)<CH3
0 N 0 CHC3H3 0 N 0 CH3
00 00 CH3 y
cH3
cH3
ci ci
General Method 7 was carried out with (7R)-11-chloro-7-(trifluoromethyl)-7,8-
dihydro-3H-
[3]benzoxocino[2,1-clpyridin-2(6H)-one (single stereoisomer) (250 mg, 758
umol, 1.0 eq.), ethyl 2-
bromo -4 -{[tert-butyl(dimethypsilyll oxy} -5,5,5 -trifluoropentanoate
(mixture of stereoisomers) (519
mg, 69% purity, 910 umol, 1.2 eq.) and 1,1,3,3-tetramethylguanidine (270 jil,
2.1 mmol, 2.8 eq.) in
a mixture of 2-propanol / acetone (4:1, 8.5 ml) including the following
variations of the procedure:
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The mixture was stirred at RT for 3 h and at 50 C overnight and then used in
the subsequent reaction
without further purification as a mixture (due to partial transesterification
during the reaction) of
ethyl
4-0- [tert-butyl (dimethypsilyll -2 -[(7R)-11 -chloro-2-oxo-7-
(trifluoromethyl) -2,6,7,8-
tetrahydro -3H-[3] benzoxocino [2, 1 -c] pyridin-3 -yl] -2,3 ,5 -trideoxy-5 ,5
,5 -trifluoropentonate (mixture
of stereoisomers):
LC-MS (method 1): R1= 1.65 / 1.64 / 1.63 min; MS (ESIpos): m/z = 642 [M+I-11+
and isopropyl 4-0- [tert-butyl (dimethypsilyll -2 -[(7R)-11 -chloro-2-oxo-7-
(trifluoromethyl) -2,6,7,8-
tetrahydro -3H-[3] benzoxocino [2, 1 -c] pyridin-3 -yl] -2,3 ,5 -trideoxy-5 ,5
,5 -trifluoropentonate (mixture
of stereoisomers):
LC-MS (method 1): R1= 1.53; MS (ESIpos): m/z = 656 [M+H1+
Example 5.20B
4-0- [tert-Butyl (dimethyl) silyll -2- [(7R)-11 -chloro-2 -oxo-7 -
(trifluoromethyl)-2,6,7,8-tetrahydro-
3H- [3] benzoxocino [2,1 -c] pyridin-3 -yll -2,3 ,5 -tride oxy-5 ,5 ,5 -
trifluoro-D-erythro-pentonic acid
(mixture of stereoisomers)
FIFj'H3 cH3
Si CH3
0'C H 3
0k 0 C H 3
N
00H
C
I
General Method 10 was carried out with a mixture of ethyl 4-0-[tert-
butyl(dimethypsily11-2-[(7R)-
11 -chloro-2-oxo-7-(trifluoromethyl) -2,6,7, 8-tetrahydro-3H-[3] benzoxocino
[2,1 -c] pyridin-3 -yl] -
2,3,5 -tride oxy-5,5,5 -trifluoropentonate (mixture of stereoisomers) and
isopropyl 4-0-[tert-
butyl (dimethyl)silyll -2- [(7R)-11 -chloro-2 -oxo -7 -(trifluoromethyl)-
2,6,7,8-tetrahydro-3H-
[3] benzoxocino [2,1 -c] pyridin-3 -yl] -2,3 ,5 -trideoxy-5 ,5 ,5 -
trifluoropentonate (mixture of
stereoisomers) (combined 350 mg, 545 lama 1.0 eq.) and lithium hydroxide (131
mg, 5.45 mmol,
10.0 eq.) including the following variations of the procedure: the crude
mixture was purified by
preparative HPLC (reversed phase, eluent: water with 0.05% formic acid /
acetonitrile 90:10 to 5:95)
leading to two separated mixtures of diastereomers.
Diastereomeric mixture 1 (desired product): 109 mg (33% of theory):
LC-MS (method 3): Rt = 4.71 / 4.73 min ; MS (ESIpos): m/z = 614 [M+I-11+ and
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Diastereomeric mixture 2: 50.0 mg (15% of theory):
LC-MS (method 3): Rt = 5.24 / 5.28 / 5.35 min ; MS (ESIpos): m/z = 614 [M+I-
11+
Example 5.20C
4-[(4-{ [tert-Butyl (dimethyl) silyll oxy } -2- [(7R)-11-chloro-2 -oxo -7 -
(trifluoromethyl)-2,6,7, 8-
tetrahydro-3H-[3]benzoxocino [2, 1 -c] pyridin-3 -yl] -5 ,5 ,5 -
trifluoropentanoyDaminolbenzamide
(mixture of stereoisomers)
F 3FC CH3
Si CH3
0' )<CH3
0 0 CH3
N
HN
0
N H2
CI 0
To a solution of 4-0-[tert-butyl(dimethypsilyll -2-[(7R)-11-chloro-2-oxo-
7-(trifluoromethyl)-
2,6,7,8-tetrahydro-3H-[3]benzoxocino [2,1 -c] pyridin-3 -yl] -2,3 ,5 -tride
oxy-5 ,5 ,5 -trifluoropentonic
acid (diastereomeric mixture 1) (15.0 mg, 24.4 umol, 1.0 eq.) in
dichloromethane (1.0 ml) was added
1-chloro-N,N,2-trimethylprop-1-en-1-amine (4.2 mg, 31.8 umol, 1.3 eq.) and the
resulting mixture
was stirred for 10 min before 4-aminobenzamide (4.3 mg, 31.8 umol, 1.3 eq.)
was added and stirring
was continued at RT for 3 h. The same procedure was repeated with 4-0-[tert-
butyl(dimethypsily11-
2-[(7R)-11-chloro-2-oxo-7-(trifluoromethyl)-2,6,7,8-tetrahydro-3H-
[3]benzoxocino [2,1 -c] pyridin-
3-y1]-2,3,5-trideoxy-5,5,5-trifluoropentonic acid (diastereomeric mixture 1)
(60.0 mg, 97.7 umol, 1.0
eq.), 1-chloro-N,N,2-trimethylprop-1-en-1-amine (17 jil, 17.0 mg, 130 umol,
1.3 eq.) and 4-
aminobenzamide (20.0 mg, 147 umol, 1.5 eq.) in dichloromethane (4.0 m1). The
two reaction
mixtures were then combined, diluted with dichloromethane, subsequently washed
with a saturated
aqueous solution of ammonium chloride and a saturated aqueous solution of
sodium bicarbonate.
The organic phase was then dried over anhydrous sodium sulfate, filtered and
concentrated under
reduced pressure. The crude mixture was used in the subsequent step without
further purification.
Yield: 88.0 mg (57% purity, 70% of theory).
LC-MS (method 3): Rt = 4.70 min; MS (ESIpos): m/z = 732 [M+I-11+
Example 5.21A
tert-Butyl (2) -2- [(7R)-11 -chloro-12 -fluoro-2-oxo-7-(trifluoromethyl)-
2,6,7,8-tetrahydro-3H-
[3] benzoxocino [2,1 -c] pyridin-3 -yl] -2,3 ,5 -trideoxy-4 -0-methyl-D-glyce
ro-pentonate (mixture of
two diastereomers)
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0,C H3
H 3
0 0 H 3
N
hC H3
0 C H 3
F
CI
General Method 7 was carried two times with (7 R) - 11-chloro-12-fluoro-7-
(trifluoromethyl)-7,8-
dihydro-3H-[3]benzoxocino[2,1-clpyridin-2(6H)-one (single stereoisomer) (130
mg, 76% purity,
284 umol, 1.0 eq.), tert-butyl (4R)-2-bromo-4-methoxypentanoate (mixture of
two diastereomers)
(98.2 mg, 85% purity, 313 umol, 1.1 eq.) and 1,1,3,3-tetramethylguanidine (107
jil, 852 umol, 3.0
eq.) in a mixture of 2-propanol / acetone (4:1, 4.5 ml) and, in a second
batch, (7R)-11-chloro-12-
fluoro-7-(trifluoromethyl)-7, 8-dihydro -3H-[3] benzoxocino [2,1 -c] pyridin-2
(6H)-one (single
stereoisomer) (100 mg, 76% purity, 219 umol, 1.0 eq.), tert-butyl (4R)-2-bromo-
4-
methoxypentanoate (mixture of two diastereomers) (75.6 mg, 85% purity, 240
umol, 1.1 eq.) and
1,1,3,3-tetramethylguanidine (82 jil, 660 umol, 3.0 eq.) in a mixture of 2-
propanol / acetone (4:1, 3.5
ml) including the following variations of the procedure: The crude mixtures of
the two reactions were
combined and purified by preparative HPLC (reversed phase, eluent: water with
0.05% formic acid
/ acetonitrile 80:20 to 5:95). Yield: 196 mg (74% of theory).
LC-MS (method 1): R1= 1.25 min; MS (ESIpos): m/z = 534 [M+H1+
Example 5.21B
(2 )-2- [(7R)-11 -Chloro -12 -fluoro-2-oxo-7-(trifluoromethyl)-2,6,7,8-
tetrahydro-3H-
[3] benzoxocino [2,1 -c] pyridin-3 -yl] -2,3 ,5 -trideoxy-4 -0-methyl-D-glyce
ro-pentonic acid (mixture
of two diastereomers)
H 3
0
H
N 0
0
0
F
CI
General Method 10 was carried out with tert-butyl (2)-2-[(7R)-11-chloro-12-
fluoro-2-oxo-7-
(trifluoromethyl)-2,6,7,8-tetrahydro-3H-[3]benzoxocino [2,1 -c] pyridin-3 -yll
-2,3,5 -trideoxy-4 -0-
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methyl-D-glycero-pentonate (mixture of two diastereomers) (192 mg, 345 umol,
1.0 eq.) and lithium
hydroxide (41.3 mg, 1.73 mmol, 5.0 eq.) in a mixture of tetrahydrofuran /
water (2:1, 13 ml) at 40 C.
Yield: 152 mg (92% of theory).
LC-MS (method 3): R1= 3.10/ 3.13 min; MS (ESIpos): m/z = 478 [M-411+
Example 5.22A
tert-Butyl (2 )-2- [(7R)-11 -chloro-12 -fluoro-2-oxo-7-(trifluoromethyl)-
2,6,7,8-tetrahydro-3H-
[3] benzoxocino [2,1 -c] pyridin-3 -yl] -2,3 ,5 -trideoxy-4 -0-methyl-L -glyce
ro -pentonate (mixture of
two diastereomers)
0'CH3
H3
0 0 H3
N
H3
00 CH3
= F
CI
General Method 7 was carried out with (7R)-11-chloro-12-fluoro-7-
(trifluoromethyl)-7,8-dihydro-
3H-[3]benzoxocino[2,1-clpyridin-2(6H)-one (single stereoisomer) (250 mg, 76%
purity, 546 umol,
1.0 eq.), tert-butyl (45)-2-bromo-4-methoxypentanoate (mixture of two
diastereomers) (161 mg, 601
umol, 1.1 eq.) and 1,1,3,3-tetramethylguanidine (210 IA, 1.6 mmol, 3.0 eq.) in
a mixture of 2-
propanol / acetone (4:1, 5 ml) including the following variations of the
procedure: The crude mixture
was purified by preparative HPLC (reversed phase, eluent: water with 0.05%
formic acid /
acetonitrile 80:20 to 5:95). Yield: 155 mg (53% of theory).
LC-MS (method 1): R1= 1.24 min; MS (ESIpos): m/z = 534 [M-411+
Example 5.22B
(2 )-2- [(7R)-11 -Chloro -12 -fluoro-2-oxo-7-(trifluoromethyl)-2,6,7,8-
tetrahydro-3H-
[3] benzoxocino [2,1 -c] pyridin-3 -yl] -2,3 ,5 -trideoxy-4 -0-methyl-L -glyce
ro -pentonic acid (mixture of
two diastereomers)
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0C H3
40C H 3
0
H
N
0
0
F
CI
General Method 10 was carried out with tert-butyl (2)-2-[(7R)-11-chloro-12-
fluoro-2-oxo-7-
(trifluoromethyl)-2,6,7,8-tetrahydro-3H-[3]benzoxocino [2,1 -c] pyridin-3 -yll
-2,3,5 -trideoxy-4 -0-
methyl-L-glycero-pentonate (mixture of two diastereomers) (155 mg, 290 umol,
1.0 eq.) and lithium
hydroxide (34.8 mg, 1.45 mmol, 5.0 eq.) in a mixture of tetrahydrofuran /
water (2:1, 4.5 m1). Yield:
185 mg (quantitative of theory).
LC-MS (method 1): Rt = 0.97 min; MS (ESIpos): m/z = 478 [M+H1+
Example 5.23A
tert-Butyl (2) -2- [(7R)-11 -chloro-12 -fluoro-2-oxo-7-(trifluoromethyl)-
2,6,7,8-tetrahydro-3H-
[3] benzoxocino [2,1 -c] pyridin-3 -yl] -2,3 ,5 -trideoxy-4 -0-
(difluoromethyl)-L -glycero-pentonate
(mixture of two diastereomers)
0 F
4 C H 3
0 0 H 3
N
H3
00 C H 3
F
CI
General Method 7 was carried out with (7 R) - 11-chloro-12-fluoro-7-
(trifluoromethyl)-7,8-dihydro-
3H-[3]benzoxocino[2,1-clpyridin-2(6H)-one (single stereoisomer) (120 mg, 76%
purity, 262 umol,
1.0 eq.), tert-butyl (4S)-2-bromo-4-(difluoromethoxy)pentanoate (mixture of
two diastereomers)
(95.4 mg, 315 umol, 1.2 eq.) and 1,1,3,3-tetramethylguanidine (99 jil, 790
umol, 3.0 eq.) in a mixture
of 2-propanol / acetone (4:1, 2.4 ml) including the following variations of
the procedure: The reaction
mixture was stirred for 4 days and the crude mixture was purified by
preparative HPLC (reversed
phase, eluent: water with 0.05% formic acid / acetonitrile 80:20 to 5:95).
Yield: 85.0 mg (53% of
theory).
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LC-MS (method 3): Rt = 4.29 min; MS (ESIpos): m/z = 570 [M+H1+
Example 5.23B
(2 )-2- [(7R)-11 -Chloro -12 -fluoro-2-oxo-7-(trifluoromethyl)-2,6,7,8-
tetrahydro-3H-
[3] benzoxocino [2,1 -c] pyridin-3 -yl] -2,3,5 -trideoxy-4 -0-(difluoromethyl)-
L -glycero-pentonic acid
(mixture of two diastereomers)
0(F
H3
0 0 H
N
0
0
F
CI
General Method 10 was carried out with tert-butyl (2)-2-[(7R)-11-chloro-12-
fluoro-2-oxo-7-
(trifluoromethyl)-2,6,7,8-tetrahydro-3H-[3]benzoxocino [2,1 -c] pyridin-3 -yl]
-2,3,5 -trideoxy-4 -0-
(difluoromethyl)-L -glycero-pentonate (mixture of two diastereomers) (83.0 mg,
94% purity, 137
lama 1.0 eq.) and lithium hydroxide (16.4 mg, 684 lama 5.0 eq.) in a mixture
of tetrahydrofuran /
water (3:1, 4 m1). Yield: 116 mg (99% of theory).
LC-MS (method 1): R1= 1.01 min; MS (ESIpos): m/z = 514 [M+I-11+
Example 6.1A
tert-Butyl [1,2,4]triazolo [1,5 -a] pyridin-7-ylcarbamate
H3C1 II
C H3 0 L m"....õ
A mixture of 7-bromo[1,2,41triazolo[1,5-alpyridine (18.5 g, 93.4 mmol, 1.0
eq.), tert-butyl
carbamate (16.42 g, 140.1 mmol, 1.5 eq.), palladium(II) acetate (2.10 g, 9.3
mmol, 0.1 eq.), 2-
dicyclohexyl-phosphino-2',4',6'-triisopropy1-1,1'-biphenyl (13.36 g, 28.0
mmol, 0.3 eq.) and caesium
carbonate (60.88 g, 186.8 mmol, 2.0 eq.) in 1,4-dioxane (930 ml) was purged
with nitrogen gas for
10 min. The reaction mixture was heated to 100 C for 15 h, cooled to RT and
filtered through Celite .
The filtrate was concentrated. The residue was purified by column
chromatography (silica gel, eluent:
petroleum ether! ethyl acetate 1:3). Yield: 17.0 g (77% of theory).
1H-NMR (400 MHz, DM50-c/6): 6 [ppm] = 9.99 (s, 1H), 8.77 (d, 1H), 8.30 (s,
1H), 7.87 (s, 1H),
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7.19-7.12 (m, 1H), 1.49 (s, 9H).
Example 6.1B
[1,2,41Triazolo[1,5-alpyridin-7-amine hydrochloride
x HCI
Hydrogen chloride solution (362.8 ml, 4 M in 1,4-dioxane) was added to a
stirred solution of tert-
butyl [1,2,41triazolo[1,5-alpyridin-7-ylcarbamate (17.00 g, 72.6 mmol) in
dichloromethane (11). The
reaction mixture was stirred at RT for 15 h and diluted with diethyl ether.
The precipitate was
collected by filtration, washed with diethyl ether and dried under reduced
pressure. Yield: 10.45 g
(84% of theory).
1H-NMR (400 MHz, DMSO-d6): 6 [ppm] = 8.97-8.90 (m, 1H), 8.67 (d, 1H), 7.40 (br
s, 2H), 6.90-
6.81 (m, 1H), 6.73-6.68 (m, 1H).
Example 6.2A
tert-Butyl (2-methyl[1,2,41triazolo[1,5-alpyridin-7-yl)carbamate
H3C1 II -CH3
CH3 0 LN..N
A mixture of 7-bromo-2-methyl[1,2,41triazolo[1,5-alpyridine (8.00 g, 37.7
mmol, 1.0 eq.), tert-butyl
carbamate (6.63 g, 56.6 mmol, 1.5 eq.), palladium(II) acetate (0.85 g, 3.8
mmol, 0.1 eq.), 2-
dicyclohexylphosphino-2',4',6'-triisopropy1-1,1'-biphenyl (5.40 g, 11.3 mmol,
0.3 eq.) and caesium
carbonate (24.58 g, 75.4 mmol, 2.0 eq.) in 1,4-dioxane (400 ml) was purged
with nitrogen for 5 min.
The reaction mixture was stirred at 100 C for 15 h, cooled to RT and filtered
through Celite . The
filtrate was concentrated under reduced pressure. The residue was purified by
column
chromatography (silica gel, eluent: petroleum ether! ethyl acetate 2:3).
Yield: 5.50 g (58% oftheory).
'H-NMR (400 MHz, DMSO-d6): 6 [ppm] = 9.92 (s, 1H), 8.63 (d, 1H), 7.74 (s, 1H),
7.12-7.03 (m,
1H), 2.37 (s, 3H), 1.49 (s, 9H).
Example 6.2B
2-Methyl[1,2,41triazolo[1,5-alpyridin-7-amine hydrochloride
H2 Nt.r.N
-CH3 x HCI
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Hydrogen chloride solution (125 ml, 4 M in 1,4-dioxane) was added to a
solution of tert-butyl (2-
methyl[1,2,41triazolo [1,5-alpyridin-7-yl)carbamate (6.20 g, 25.0 mmol) in
dichloromethane
(125 m1). The reaction mixture was stirred at RT for 15 h and diluted with
diethyl ether. The
precipitate was collected by filtration, washed with diethyl ether and dried
under reduced pressure.
Yield: 3.69 g (78% of theory).
'H-NMR (400 MHz, DMSO-d6): 6 [ppm] = 8.58 (d, 1H), 7.45 (br s, 2H), 6.82 (dd,
1H), 6.66 (d, 1H),
2.49 (s, 3H).
Example 6.3A
1,1-Diphenyl-N-( [1,2,4]triazolo [1,5 -alpyridin-6-yl)methanimine
1101 N
kN
A mixture of 6-bromo[1,2,41triazolo[1,5-alpyridine (3.00 g, 15.2 mmol), 1,1-
diphenylmethanimine
(5.49 g, 30.3 mmol, 2.0 eq.), tris(dibenzylideneacetone)dipalladium (1.11 g,
1.2 mmol, 0.08 eq.),
2,2'-bis(diphenylphosphino)-1,1'-binaphthyl (1.89 g, 3.0 mmol, 0.2 eq.) and
sodium tert-butoxide
(4.37 g, 45.5 mmol, 3.0 eq.) in toluene (100 ml) was heated at 80 C under
nitrogen atmosphere
overnight. The reaction mixture was cooled to RT and concentrated under
reduced pressure. The
residue was poured into water and extracted with dichloromethane. The combined
organic phases
were dried over anhydrous sodium sulfate, filtered and concentrated under
reduced pressure. The
residue was purified by column chromatography (silica gel, eluent: petroleum
ether / ethyl acetate
3:1). Yield: 3.00 g (64% of theory).
'H-NMR (300 MHz, DMSO-d6): 6 [ppm] = 8.34 (d, 1H), 7.74-7.41 (m, 6H), 7.38-
7.13 (m, 6H).
Example 6.3B
[1,2,4] Triazolo [1,5 -a] pyridin-6-amine
H2N
NN
Aqueous hydrochloric acid (50 ml, 2 N, 101 mmol, 10 eq.) was added to a
solution of 1,1-diphenyl-
N-([1,2,41triazolo[1,5-alpyridin-6-yl)methanimine (3.00 g, 10.1 mmol) in
tetrahydrofuran (100 ml).
The reaction mixture was stirred at RT for 2 h and washed with ethyl acetate.
The aqueous phase was
adjusted to neutral with aqueous solution of sodium hydroxide (0.1 M) and
concentrated under
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reduced pressure. The residue was purified by column chromatography (reversed
phase, eluent:
water! acetonitrile: 10% acetonitrile to 35% acetonitrile in 15 min, flow
rate: 80 ml/min). Yield: 1.10
g (81% of theory).
'H-NMR (400 MHz, DMSO-d6): 6 [ppm] = 8.17 (s, 1H), 8.03 (d, 1H), 7.56 (d, 1H),
7.20 (d, 1H),
5.25 (br s, 2H).
Example 6.4A
6-Bromo [1,2,4]triazolo [4,3 -a] pyridin-3 (2H)-one
0
Br
NH
5-Bromo-2-hydrazinopyridine (15.00 g, 79.78 mmol, 1.0 eq.) was added in
several portions over 0.5
h at 0 C to a solution of bis(trichloromethyl) carbonate (71.02 g, 239.3 mmol,
3.0 eq.) in
tetrahydrofuran (11). After stirring for 15 h at RT, the reaction mixture was
diluted with water,
adjusted to pH 8 with sodium carbonate and extracted with ethyl acetate. The
combined organic
phases were washed with brine, dried over anhydrous sodium sulfate, filtered
and evaporated to
dryness. Yield: 11.30 g (62% of theory).
'H-NMR (400 MHz, DMSO-d6): 6 [ppm] = 12.60 (s, 1H), 8.06 (t, 1H), 7.25-7.19
(m, 2H).
Example 6.4B
6-Bromo-2-methyl [1,2,4]triazolo [4,3 -a] pyridin-3 (211)-one
0
Br k
N - C H 3
Iodomethane (5.73 g, 40.4 mmol, 1.2 eq.) was added dropwise at RT to a mixture
of 6-
bromo[1,2,41triazolo[4,3-alpyridin-3(211)-one (7.20 g, 33.6 mmol, 1.0 eq.) and
caesium carbonate
(12.06 g, 37.0 mmol, 1.1 eq.) in /V,N-dimethylformamide (144 m1). After
stirring for 2 h, the reaction
mixture was concentrated under reduced pressure. The residual mixture was
diluted with water and
extracted with ethyl acetate. The combined organic phases were washed with
brine, dried over
anhydrous sodium sulfate, filtered and concentrated under reduced pressure.
The residue was purified
by column chromatography (silica gel, eluent: petroleum ether / ethyl acetate
1:1). Yield: 7.10 g
(92% of theory).
'H-NMR (400 MHz, DMSO-d6): 6 [ppm] = 8.13-8.09 (m, 1H), 7.28 (dd, 1H), 7.22
(dd, 1H), 3.52 (s,
3H).
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Example 6.4C
tert-Butyl (2-methyl-3-oxo-2,3-dihydro [1,2,4]triazolo [4,3 -a] pyridin-6 -
yl)carbamate
0
HH:CC>rOY N H3
CH3 0 L,LN\
A mixture of 6-bromo-2-methyl[1,2,41triazolo[4,3-alpyridin-3(2H)-one (3.00 g,
13.2 mmol, 1.0 eq.),
tert-butyl carbamate (2.31 g, 19.7 mmol, 1.5 eq.),
tris(dibenzylideneacetone)dipalladium chloroform
complex (0.68 g, 0.7 mmol, 0.05 eq.), 9,9-dimethy1-4,5-
bis(diphenylphosphino)xanthene (1.14 g,
2.0 mmol, 0.15 eq.) and caesium carbonate (8.57 g, 26.3 mmol, 2.0 eq.). in 1,4-
dioxane (130 ml) was
purged with nitrogen for 5 minutes. The reaction was stirred at 110 C for 4 h,
cooled to RT and
filtered through Celite . The filtrate was concentrated under reduced
pressure. The residue was
purified by column chromatography (silica gel, eluent: petroleum ether / ethyl
acetate 1:4). Yield:
3.00 g (82% of theory).
'H-NMR (300 MHz, DMSO-d6): 6 [ppm] = 9.43 (br s, 1H), 8.08 (br s, 1H), 7.20
(dd, 1H), 7.13 (dd,
1H), 3.48 (s, 3H), 1.44 (s, 9H).
Example 6.4D
6-Amino -2 -methyl [1,2,4]triazolo [4,3 -a] pyridin-3 (211)-one hydrochloride
0
H2Nr.Ncl4
N-CH3 x HCI
Hydrogen chloride solution (100 ml, 4 M in 1,4-dioxane) was added to a
solution of tert-butyl (2-
methy1-3-oxo-2,3-dihydro [1,2,41triazolo[4,3-alpyridin-6-yl)carbamate (6.50 g,
22.4 mmol) in
dichloromethane (100 m1). The reaction mixture was stirred for 6 h at RT and
diluted with
dichloromethane. The precipitate was collected by filtration, washed with
acetonitrile and dried in
vacuo. Yield: 3.26 g (70% of theory).
'H-NMR (300 MHz, DMSO-d6): 6 [ppm] = 9.05 (br s, 3H), 7.71 (s, 1H), 7.27 (dd,
1H), 7.09 (dd,
1H), 3.50 (s, 3H).
Example 6.5A
4-Bromo-2-hydrazinopyridine
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N H 2
Br NH
N
Hydrazine hydrate (80.0 g, 80% in water, 1.3 mol, 15.0 eq.) was added to a
solution of 4-bromo-2-
fluoropyridine (15.0 g, 85.2 mmol) in ethanol (200 m1). The reaction mixture
was stirred at RT for
15 h and concentrated under reduced pressure. The residual mixture was diluted
with water and
.. extracted with ethyl acetate. The combined organic phases were washed with
brine, dried over
anhydrous sodium sulfate, filtered and evaporated to dryness. Yield: 15.0 g
(93% of theory).
1H-NMR (400 MHz, DMSO-d6): 6 [ppm] = 7.85 (d, 1H), 7.72 (s, 1H), 6.93 (d, 1H),
6.72-6.70 (m,
1H), 4.21 (s, 2H).
Example 6.5B
.. 7-Bromo [1,2,4]triazolo [4,3 -a] pyridin-3 (211)-one
Br
N H
0
4-Bromo-2-hydrazinopyridine (10.0 g, 53.2 mmol) was added in several portions
over 0.5 h at 0 C
to a solution of 1,1'-carbonyldiimidazole (17.25 g, 106.4 mmol, 2.0 eq.) in
tetrahydrofuran (600 m1).
After stirring for 15 h at RT, the reaction mixture was diluted with saturated
sodium bicarbonate
solution and extracted with ethyl acetate. The combined organic phases were
washed with brine,
dried over anhydrous sodium sulfate, filtered and concentrated under reduced
pressure. The residue
was purified by column chromatography (silica gel, eluent: dichloromethane /
methanol 50:1). Yield:
7.3 g (63% of theory).
1H-NMR (400 MHz, DMSO-d6): 6 [ppm] = 12.55 (s, 1H), 7.79-7.76 (m, 1H), 7.63
(d, 1H), 6.68-6.66
(m, 1H).
Example 6.5C
7-Bromo-2-methyl [1,2,4]triazolo [4,3 -a] pyridin-3 (211)-one
Br
N-C H 3
0
Iodomethane (4.77 g, 33.6 mmol, 1.2 eq.) was added dropwise at RT to a mixture
of 7-
bromo[1,2,41triazolo[4,3-alpyridin-3(211)-one (6.00 g, 28.0 mmol) and caesium
carbonate (13.70 g,
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42.0 mmol, 1.5 eq.) in NN-dimethylformamide (100 m1). After stirring for 3 h
at RT, the reaction
mixture was concentrated under reduced pressure. The residual mixture was
diluted with water and
extracted with ethyl acetate. The combined organic layers were washed with
brine, dried over
anhydrous sodium sulfate, filtered and concentrated under reduced pressure.
The residue was purified
by column chromatography (silica gel, eluent: petroleum ether / ethyl acetate
1:1). Yield: 3.60 g
(56% of theory).
1H-NMR (300 MHz, DMSO-d6): 6 [ppm] = 7.83-7.80 (m, 1H), 7.66-7.65 (m, 1H),
6.73-6.70 (m,
1H), 3.52 (s, 3H).
Example 6.5D
tert-Butyl (2-methyl-3-oxo-2,3-dihydro [1,2,4]triazolo [4,3 -a] pyridin-7 -
yl)carbamate
H3C0 N
II
Cr-- =
H3Ci N-C H3
CH3 0 \
0
tert-Butyl carbamate (2.77 g, 23.7 mmol, 1.5 eq.),
tris(dibenzylideneacetone)dipalladium chloroform
complex (0.82 g, 0.8 mmol, 0.05 eq.), 9,9-dimethy1-4,5-bis(diphenylphosphino)-
xanthene (1.37 g,
2.4 mmol, 0.15 eq.) and caesium carbonate (10.29 g, 31.6 mmol, 2.0 eq.) were
added to a solution of
7-bromo-2-methyl[1,2,41triazolo[4,3-alpyridin-3(211)-one (3.60 g, 15.8 mmol)
in 1,4-dioxane (72
m1). The resulting mixture was purged with nitrogen for 5 minutes, stirred at
110 C for 4 h, cooled
to RT and filtered through Celite . The filtrate was concentrated under
reduced pressure. The residue
was purified by chromatography (silica gel, eluent: petroleum ether / ethyl
acetate 1:4). Yield: 3.10 g
(68% of theory).
1H-NMR (300 MHz, DMSO-d6): 6 [ppm] = 9.82 (m, 1H), 7.79-7.76 (m, 1H), 7.28-
7.27 (m, 1H),
6.58-6.55 (m, 1H), 3.45 (s, 3H), 1.48 (s, 9H).
Example 6.5E
7-Amino -2 -methyl [1,2,4]triazolo [4,3 -a] pyridin-3 (211)-one hydrochloride
H2 N N
N-CH3 x HCI
N,\<
0
Hydrogen chloride solution (100 ml, 4 M in 1,4-dioxane) was added to a
solution of tert-butyl (2-
methy1-3-oxo-2,3-dihydro [1,2,41triazolo[4,3-alpyridin-7-yl)carbamate (5.70 g,
19.8 mmol) in
dichloromethane (100 m1). The reaction mixture was stirred for 6 h at RT and
diluted with
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dichloromethane. The precipitate was collected by filtration, washed with
acetonitrile and dried in
vacuo. Yield: 3.06 g (76% of theory).
1H-NMR (300 MHz, DMSO-d6): 6 [ppm] = 7.59-7.56 (m, 1H), 6.28-6.25 (m, 2H),
6.20-6.17 (m,
2H), 5.76 (d, 1H), 3.36 (s, 3H).
Example 6.6A
tert-Butyl [1,2,4]triazolo [4,3 -alpyridin-7-ylcarbamate
H 3C 0 N
H3C1 Y = C H3 0
tert-Butyl carbamate (8.87 g, 75.7 mmol, 3.0 eq.),
tris(dibenzylideneacetone)dipalladium chloroform
complex (2.61 g, 2.5 mmol, 0.1 eq.), 9,9-dimethy1-4,5-
bis(diphenylphosphino)xanthene (4.38 g, 7.6
mmol, 0.3 eq.) and caesium carbonate (16.4 g, 50.5 mmol, 2.0 eq.) were added
to a solution of 7-
bromo[1,2,41triazolo[4,3-alpyridine (5.00 g, 25.2 mmol) in 1,4-dioxane (250
m1). The resulting
mixture was purged with nitrogen for 5 minutes, stirred at 110 C for 4 h,
cooled to RT and filtered
through Celite . The filtrate was concentrated under reduced pressure. The
residue was purified by
chromatography (silica gel, eluent: petroleum ether / ethyl acetate 1:9).
Yield: 3.50 g (49% of
theory).
1H-NMR (300 MHz, DMSO-d6): 6 [ppm] = 9.88 (s, 1H), 9.06 (s, 1H), 8.42 (d, 1H),
7.80 (s, 1H),
6.97-6.94 (m, 1H), 1.51 (s, 9H).
Example 6.6B
[1,2,41Triazolo[4,3-al pyridin-7-amine hydrochloride
H2Nor...N
N x HCI
Hydrogen chloride solution (107 ml, 4 M in 1,4-dioxane) was added to a stirred
solution of tert-butyl
[1,2,41triazolo[4,3-alpyridin-7-ylcarbamate (5.00 g, 21.3 mmol) in
dichloromethane (120 ml). The
reaction mixture was stirred for 15 h at RT and diluted with dichloromethane.
The precipitate was
collected by filtration, washed with dichloromethane and dried in vacuo. The
crude product was
purified by flash chromatography (330 g C18-bonded silica gel, 100% water).
Yield: 2.40 g (58% of
theory).
1H-NMR (300 MHz, DMSO-d6): 6 [ppm] = 9.15 (s, 1H), 8.47-8.44 (m, 1H), 7.62 (s,
2H), 6.92-6.70
(m, 1H), 6.58 (d, 1H).
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Example 6.7A
tert-Butyl (3-methyl [1,2,4] triazolo [4,3 -a] pyridin-7-yl)carbamate
H
H3COIINrr...N
H3C1
CH3
CH3
A mixture of 7-bromo-3-methyl[1,2,41triazolo[4,3-a]pyridine (210 mg, 0.99
mmol), tert-butyl
carbamate (174 mg, 1.49 mmol, 1.5 eq.), palladium(II) acetate (22 mg, 0.1
mmol, 0.1 eq.), 2-
dicylohexylphosphino-2' ,4' ,6' -triisopropylbiphenyl (142 mg, 0.3 mmol, 0.3
eq.) and caesium
carbonate (647 mg, 1.98 mmol, 2.0 eq.). in 1,4-dioxane (10 ml) was stirred at
100 C overnight.
Tris(dibenzylideneacetone)dipalladium (18 mg, 0.02 mmol, 0.02 eq.) and 9,9-
dimethy1-4,5-
bis(diphenylphosphino)xanthene (46 mg, 0.08 mmol, 0.08 eq.) were added. The
reaction mixture
was stirred at 100 C for additional 3 h, cooled to RT and filtered through
Celite . The filtrate was
concentrated under reduced pressure. The residue was purified by preparative
HPLC (reversed phase,
eluent: acetonitrile / water gradient). Yield: 72 mg (29% of theory).
LC-MS (method 4): Rt = 0.86 min; MS (ESIpos): m/z = 249 [M+H1+
Example 6.7B
3-Methyl [1,2,4]triazolo [4,3-al pyridin-7 -amine hydrochloride
....crH2N .....õ N,
N x HCI
N.....j(
CH3
A solution of tert-butyl (3-methyl[1,2,41triazolo[4,3-alpyridin-7-yl)carbamate
(72 mg, 0.29 mmol)
in hydrogen chloride solution (1.44 ml, 4 M in 1,4-dioxane, 5.77 mmol, 20 eq.)
was stirred at RT
overnight. Additional hydrogen chloride solution (1.44 ml, 4 M in 1,4-dioxane,
5.77 mmol, 20 eq.)
was added. The reaction mixture was stirred again at RT overnight,
concentrated in vacuo,
coevaporated two times with dichloromethane and used in the subsequent
reaction without further
purification. Yield: 59 mg (quantitative of theory).
LC-MS (method 15): Rt = 0.43 min; MS (ESIpos): m/z = 149 [M+I-11+
'H-NMR (500 MHz, DM50-d6): 6 [ppm] = 8.37 (d, 1H), 7.55 (br s, 2H), 6.89 (dd,
1H), 6.53 (d, 1H),
2.61 (s, 3H).
Example 6.8A
tert-Butyl [1,2,4]triazolo [4,3 -alpyridin-6-ylcarbamate
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H
H 3C0.(NONI...7
H3C" I II
C H3 0 \
A mixture of 6-bromo[1,2,41triazolo[4,3-alpyridine (1.00 g, 5.05 mmol), tert-
butyl carbamate
(887 mg, 7.58 mmol, 1.5 eq.), tris(dibenzylideneacetone)dipalladium (93 mg,
0.10 mmol, 0.02 eq.),
9,9-dimethy1-4,5-bis(diphenylphosphino)xanthene (234 mg, 0.40 mmol, 0.08 eq.)
and caesium
carbonate (3.29 g, 10.10 mmol, 2.0 eq.). in 1,4-dioxane (20 ml) was stirred at
100 C overnight.
Tris(dibenzylideneacetone)dipalladium (46 mg, 0.05 mmol, 0.01 eq.) and 9,9-
dimethy1-4,5-
bis(diphenylphosphino)xanthene (117 mg, 0.20 mmol, 0.04 eq.) were added. The
reaction mixture
was stirred again at 100 C overnight, cooled to RT and filtered through Celite
. The filtrate was
concentrated under reduced pressure. The residue was purified by column
chromatography (silica
gel, eluent: dichloromethane / methanol gradient). Yield: 167 mg (14% of
theory).
LC-MS (method 4): R1= 1.09 min; MS (ESIpos): m/z = 235 [M+H1+
'H-NMR (600 MHz, DM50-d6): 6 [ppm] = 9.68 (br s, 1H), 9.23 (d, 1H), 8.97 (br
s, 1H), 7.73 (d,
1H), 7.26 (dd, 1H), 1.50 (s, 9H).
Example 6.8B
[1,2,4]Triazolo [4,3 -a] pyridin-6-amine hydrochloride
H2NoNo
x HCI
A solution of tert-butyl [1,2,41triazolo[4,3-alpyridin-6-ylcarbamate (180 mg,
0.75 mmol) in
hydrogen chloride solution (3.77 ml, 4 M in 1,4-dioxane, 15.09 mmol, 20 eq.)
was stirred at RT
overnight, concentrated in vacuo, coevaporated two times with dichloromethane
and used in the
subsequent reaction without further purification. Yield: 171 mg (80% purity).
'H-NMR (600 MHz, DM50-d6): 6 [ppm] = 9.53 (s, 1H), 8.00 (d, 1H), 7.90 (d, 1H),
7.66 (dd, 1H),
6.0 (br s, 3H).
Workin2 examples
Example 1
4-{ R25)-2 -(11-Chloro -7 -methy1-2 -oxo -2,6,7, 8-tetrahydro-3H-
[3]benzoxocino [2,1-c] pyridin-3 -
yl)butanoyllamino}-2-fluorobenzamide (mixture of two diastereomers)
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C H 3
0
HH 3C
0 0
0
N H 2
CI
1,1,3,3-Tetramethylguanidine (52
0.42 mmol, 3.0 eq.) was added under argon atmosphere at RT
to a solution of 11-chloro-7-methyl-7,8-dihydro-3H-[3]benzoxocino[2,1-
c]pyridin-2(6H)-one
(racemate) (40 mg, 0.14 mmol) in 2-propanol / acetone (4:1, 0.5 m1). The
mixture was stirred at RT
for 15 min, followed by addition of 4-{[(2R)-2-bromobutanoyllamino}-2-
fluorobenzamide (single
stereoisomer) (63 mg, 0.21 mmol, 1.5 eq.) and further 2-propanol / acetone
(4:1, 0.6 m1). The
reaction mixture was stirred at RT overnight and concentrated under reduced
pressure. The residue
was purified by preparative HPLC (column: Chromatorex C18 10 pm, 125 mm x 30
mm; eluent:
acetonitrile / 0.05% formic acid; gradient: 0-3 min 10% acetonitrile, to 35
min 90% acetonitrile and
further 3 min 90% acetonitrile). Yield: 58 mg (83% of theory).
LC-MS (method 4): R1= 1.88 min; MS (ESIpos): m/z = 498 [M-411+
'H-NMR (500 MHz, DM50-d6): 6 [ppm] = 10.91-10.81 (m, 1H), 7.83-7.75 (m, 1H),
7.73-7.62 (m,
2H), 7.59-7.48 (m, 2H), 7.48-7.30 (m, 4H), 6.42-6.32 (m, 1H), 5.61-5.51 (m,
1H), 4.54-4.44 / 4.34-
4.25 / 3.91-3.81 / 3.39-3.26 (4m, 2H, partially concealed), 2.91-2.80 /2.75-
2.60 (2m, 1H), 2.35-2.25
/ 2.22-1.95 (2m, 4H), 0.98-0.83 (m, 6H). Additional signals of minor rotamers
were also detected.
Example 2
4-(1(25)-24(7R)-11-Chloro-7-methyl-2-oxo-2,6,7,8-tetrahydro-3H-[3]benzoxocino
[2,1-clpyridin-
3 -yll butanoyl } amino)-2-fluorobenzamide (single stereoisomer)
C H3
0
EN-11
H 3C
0 1401 0
0
N H
CI
Diastereomer separation of 58 mg of 4-{ [(2S)-2-(11-chloro-7-methy1-2-oxo-
2,6,7,8-tetrahydro-3H-
[3] benzoxocino [2,1-c] pyridin-3 -yl)butanoyll amino -2-fluorobenzamide
(mixture of two
diastereomers), Example 1 gave
single stereoisomer 1 (chiral SFC: Rt = 0.97 min, >99% de): 20 mg,
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single stereoisomer 2 (chiral SFC: R1= 1.98 min, >99% de): 21 mg.
Separation method: SFC: column: Daicel Chiralpak AD, 250 mm x 20 mm; eluent:
72% carbon
dioxide /28% 2-propanol; temperature: 40 C; flow rate: 80 ml/min; UV
detection: 210 nm.
Analysis method: SFC: column: Daicel Chiralpak AD, 50 mm x 4.6 mm; eluent: 70%
carbon
.. dioxide / 30% 2-propanol; temperature: 40 C; flow rate: 3 ml/min; UV
detection: 210 nm.
20 mg of single stereoisomer 1 were further purified by preparative HPLC
(column: Chromatorex
C18 10 um, 125 mm x 30 mm; eluent: acetonitrile / 0.05% formic acid; gradient:
0-3 min 10%
acetonitrile, to 35 min 90% acetonitrile and further 3 min 90% acetonitrile)
to give 18 mg of the title
compound Example 2.
LC-MS (method 4): R1= 1.88 min; MS (ESIpos): m/z = 498 [M+H1+
'H-NMR (500 MHz, DM50-d6): 6 [ppm] = 10.90-10.82 (m, 1H), 7.78 (s, 1H), 7.70
(t, 1H), 7.68-
7.61 (m, 1H), 7.58-7.49 (m, 2H), 7.48-7.36 (m, 3H), 7.36-7.30 (m, 1H), 6.41-
6.32 (m, 1H), 5.60-5.52
(m, 1H), 4.54-4.46 / 4.32-4.25 / 3.90-3.83 / 3.3-3.24 (4m, 2H, partially
concealed), 2.90-2.81 / 2.76-
2.62 (2m, 1H, partially concealed), 2.34-2.26 / 2.22-1.96 (2m, 4H), 0.97-0.83
(m, 6H). Additional
signals of minor rotamers were also detected.
Example 3
4-( 1(25)-2 -R7R)- 11 -Chloro-7-methy1-2-oxo-2,6,7,8 -tetrahydro-3H-
[3]benzoxocino [2,1-c] pyridin-
3 -yll butanoyl} amino)benzamide (single stereoisomer)
C H 3
0 EI
H 3C
N N
0
NH
2
0
Ci
1,1,3,3-Tetramethylguanidine (38 jil, 0.30 mmol, 3.0 eq.) was added under
argon atmosphere at RT
to a solution of (7 R) - 11-chloro-7-methyl-7,8-dihydro-3H-[3]benzoxocino[2,1-
c]pyridin-2(6H)-one
(single stereoisomer) (32 mg, 86% purity, 0.10 mmol) in 2-propanol / acetone
(4:1, 1.25 m1). The
mixture was stirred at RT for 15 min, followed by addition of 4-{[(2R)-2-
bromobutanoyllamino}benzamide (single stereoisomer) (31 mg, 0.11 mmol, 1.1
eq.). The reaction
mixture was stirred at RT overnight and concentrated under reduced pressure.
The residue was
purified by preparative HPLC (reversed phase, eluent: acetonitrile / water
gradient). Yield: 26 mg
(54% of theory).
LC-MS (method 4): R1= 1.79 min; MS (ESIpos): m/z = 480 [M+H1+
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'H-NMR (400 MHz, DMSO-d6): 6 [ppm] = 10.69 (br s, 1H), 7.92-7.77 (m, 4H), 7.73-
7.63 (m, 2H),
7.50-7.30 (m, 3H), 7.30-7.20 (m, 1H), 6.42-6.32 (m, 1H), 5.66-5.55 (m, 1H),
4.55-4.45 /4.33-4.24 /
3.90-3.82 / 3.3-3.23 (4m, 2H, partially concealed), 2.91-2.81 / 2.76-2.63 (2m,
1H), 2.35-2.25 / 2.22-
1.96 (2m, 4H), 0.98-0.83 (m, 6H). Additional signals of minor rotamers were
also detected.
Example 4
(25)-2 -[(7R)-11 -Chloro-7-methyl-2 -oxo -2,6,7, 8-tetrahydro-3H- [3]
benzoxocino [2,1 -c] pyridin-3 -yll -
N-(2-methy1-2H-benzotriazol -5-yl)butanamide (single stereoisomer)
C H 3
ENI
H3 C
N
0 Th\l/NI¨C H 3
0
=
C I
1,1,3,3-Tetramethylguanidine (38 0.30 mmol, 3.0 eq.) was added under argon
atmosphere at RT
to a solution of (7 R) - 11-chloro-7-methyl-7,8-dihydro-3H-[3]benzoxocino[2,1-
c]pyridin-2(6H)-one
(single stereoisomer) (32 mg, 86% purity, 0.10 mmol) in 2-propanol / acetone
(4:1, 1.25 m1). The
mixture was stirred at RT for 15 min, followed by addition of (2R)-2-bromo-N-
(2-methy1-2H-
benzotriazol-5-yl)butanamide (single stereoisomer) (31 mg, 0.10 mmol, 1.0
eq.). The reaction
mixture was stirred at RT overnight and concentrated under reduced pressure.
The residue was
purified by preparative HPLC (reversed phase, eluent: acetonitrile / water
gradient). Yield: 38 mg
(77% of theory).
LC-MS (method 4): Rt = 2.06 min; MS (ESIpos): m/z = 492 [M+H1+
'H-NMR (600 MHz, DMSO-d6): 6 [ppm] = 10.76-10.69 (m, 1H), 8.38-8.33 (m, 1H),
7.89 (d, 1H),
7.85-7.80 (m, 1H), 7.51-7.31 (m, 4H), 6.42-6.33 (m, 1H), 5.68-5.60(m, 1H),
4.54-4.48 / 4.32-4.27 /
3.91-3.84 / 3.37-3.3 (4m, 2H, partially concealed), 4.45 (s, 3H), 2.89-2.82
/2.75-2.64 (2m, 1H), 2.35-
2.27 / 2.23-1.98 (2m, 4H), 0.98-0.84 (m, 6H). Additional signals of minor
rotamers were also
detected.
Example 5
4-( { (25)-2 -R7R)- 11 -Chloro-7-methy1-2-oxo-2,6,7,8 -tetrahydro-3H-
[3]benzoxocino [2,1-clpyridin-
3 -yl] -3 - [(25)-tetrahydro-2H-pyran-2 -yll propanoyllamino)-2 -
fluorobenzamide (single stereoisomer)
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0
H 3C
N
00 .1
NH
2
0
CI
1,1,3,3-Tetramethylguanidine (38 0.30 mmol, 3.0 eq.) was added under argon
atmosphere at RT
to a solution of (7 R) - 11-chloro-7-methyl-7,8-dihydro-3H-[3]benzoxocino[2,1-
c]pyridin-2(6H)-one
(single stereoisomer) (32 mg, 86% purity, 0.10 mmol) in 2-propanol / acetone
(4:1, 1.25 m1). The
mixture was stirred at RT for 15 min, followed by addition of 4-({(2R)-2-bromo-
3-[(25)-tetrahydro-
2H-pyran-2-yllpropanoyl}amino)-2-fluorobenzamide (single stereoisomer) (37 mg,
0.10 mmol, 1.0
eq.). The reaction mixture was stirred at RT overnight and concentrated under
reduced pressure. The
residue was purified by preparative HPLC (reversed phase, eluent: acetonitrile
/ water gradient).
Yield: 27 mg (47% of theory).
LC-MS (method 4): Rt = 2.05 min; MS (ESIpos): m/z = 568 [M+H1+
'H-NMR (400 MHz, DM50-d6): 6 [ppm] = 10.85-10.66 (m, 1H), 7.84-7.76 (m, 1H),
7.72-7.62 (m,
2H), 7.58-7.48 (m, 2H), 7.48-7.37 (m, 3H), 7.36-7.29 (m, 1H), 6.38-6.30 (m,
1H), 5.81-5.65 (m, 1H),
4.54-4.44 /4.31-4.23 / 3.92-3.79 /3.3-3.16 (4m, 5H, partially concealed), 2.90-
2.80 / 2.75-2.65 (2m,
1H), 2.35-2.19 / 2.17-1.95 (2m, 4H), 1.82-1.70 (m, 1H), 1.67-1.57 (m, 1H),
1.49-1.34 (m, 3H), 1.33-
1.20 (m, 1H), 0.96-0.82 (m, 3H). Additional signals of minor rotamers were
also detected.
Example 6
4-( 1(25)-2 -R7R)- 11 -Chloro-7-methy1-2-oxo-2,6,7,8 -tetrahydro-3H-
[3]benzoxocino [2,1-c] pyridin-
3 -yll -3 - [(25)-tetrahydro-2H-pyran-2 -yll propanoyllamino)benzamide (single
stereoisomer)
0
0 c111(1
HG
N
00 N H 2
0
CI
General Method 7 was carried out with (7 R) - 11-chloro-7-methy1-7,8 -dihydro -
3H-
[3]benzoxocino [2,1-c] pyridin-2(6H)-one (single stereoisomer) (50.0 mg, 181
lama 1.0 eq.), 4-
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({(2R)-2-bromo-3-[(25)-tetrahydro-2H-pyran-2-yllpropanoyl}amino)benzamide
(single
stereoisomer) (96.6 mg, 272 lama 1.5 eq.) and 1,1,3,3-tetramethylguanidine (68
1.1.1, 540 lama 3.0
eq.) in a mixture of 2-propanol / acetone (4:1, 2.6 m1). The crude mixture was
purified by preparative
HPLC (reversed phase, eluent: water with 0.05% formic acid! acetonitrile 80:20
to 1:99). Yield: 50.0
mg (50% of theory).
LC-MS (method 1): R1= 1.04 min; MS (ESIpos): m/z = 550 [M+H1+
1H-NMR (500 MHz, DM50-c/6): 6 [ppm] = 10.70! 10.63 (2s, 1H), 7.90 (br s, 1H),
7.87-7.80 (m,
3H), 7.73-7.66 (m, 2H), 7.49-7.38 (m, 2H), 7.35-7.31 (m, 1H), 7.29 (br s, 1H),
6.37! 6.32 (2s, 1H),
5.81 / 5.75 (2t, 1H), 4.54-4.44 (m, 1H), 3.91-3.80 (m, 1H), 3.31-3.18 (m, 3H),
2.70 (d, 1H), 2.36-
2.17 (m, 2H), 2.17-2.07 (m, 1H), 2.06-1.95 (m, 1H), 1.80-1.71 (m, 1H), 1.67-
1.56 (m, 1H), 1.46-1.34
(m, 3H), 1.32-1.17 (m, 1H), 0.91! 0.86 (2d, 3H). Additional signals of minor
rotamers were also
detected.
Example 7
4-{2-[(7R)-11-Chloro -7-methy1-2-oxo-2,6,7,8 -tetrahydro-3H-[3]benzoxocino
[2,1-clpyridin-3 -yl] -
3-{(2R)-1,4-dioxan-2-yllpropanamido}benzamide (mixture of two diastereomers)
C)
0
0
H 3C
N
00 N H 2
CI
2-[(7R)-11 -Chloro -7-methy1-2-oxo-2,6,7,8 -tetrahydro-3H-[3]benzoxocino [2,1-
c] pyridin-3 -yl] -3 -
[(2R)-1,4-dioxan-2-yllpropanoic acid (mixture of two diastereomers) (51 mg,
0.12 mmol) and 4-
aminobenzamide (24 mg, 0.18 mmol, 1.5 eq.) were reacted according to General
Method 11. Yield:
57 mg (86% of theory).
LC-MS (method 4): R1= 1.65 / 1.68 min; MS (ESIpos): m/z = 552 [M+H1+
1H-NMR (400 MHz, DM50-c/6): 6 [ppm] = 10.73-10.56 (m, 1H), 7.91-7.77 (m, 4H),
7.73-7.64 (m,
2H), 7.48-7.30 (m, 3H), 7.29-7.20 (m, 1H), 6.40-6.31 (m, 1H), 5.81-5.66 (m,
1H), 4.53-4.44 / 4.34-
4.24 / 3.92-3.81 / 3.40-3.19 (4m, 3H, partially concealed), 3.77-3.57 (m, 3H),
3.55-3.40 (m, 3H),
2.90-2.82 / 2.76-2.65 (2m, 1H), 2.35-1.95 (m, 4H), 0.96-0.83 (m, 3H).
Additional signals of minor
rotamers were also detected.
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Example 8
4-( (25)-2 -R7R)- 11 -Chloro-7-methy1-2-oxo-2,6,7,8 -tetrahydro-3H-
[3]benzoxocino [2,1-clpyridin-
3 -yll -3 - [(2R)-1,4 -dioxan-2 -yll propanoyllamino)benzamide (single
stereoisomer)
IH
H 3C
N
00J NH 2
0
CI
Diastereomer separation of 50 mg of 4-{2-[(7R)-11-chloro-7-methy1-2-oxo-
2,6,7,8-tetrahydro-3H-
[3] benzoxocino [2,1-c] pyridin-3 -yl] -3 -{(2R)-1,4 -dioxan-2 -yll
propanamidolbenzamide (mixture of
two diastereomers), Example 7 gave
single stereoisomer 1 (chiral HPLC: Rt = 3.58 min, 99% de): 18 mg,
single stereoisomer 2 (chiral HPLC: Rt = 4.60 min, 99% de): 37 mg.
Separation method: HPLC: column: Daicel Chiralcel OX-H 5 um, 250 mm x 20 mm;
eluent: 50%
n-heptane / 50% ethanol; temperature: 40 C; flow rate: 15 ml/min; UV
detection: 210 nm.
Analysis method: HPLC: column: Daicel Chiralpak OX-3 3 um, 50 mm x 4.6 mm;
eluent: 50%
n-heptane / 50% ethanol; temperature: 40 C; flow rate: 1 ml/min; UV detection:
220 nm.
18 mg of single stereoisomer 1 were further purified by preparative HPLC
(reversed phase, eluent:
acetonitrile / 0.05% formic acid gradient) to give 15 mg of the title compound
Example 8.
LC-MS (method 4): R1= 1.69 min; MS (ESIpos): m/z = 552 [M-411+
'H-NMR (500 MHz, DM50-d6): 6 [ppm] = 10.69-10.55 (m, 1H), 7.91-7.76 (m, 4H),
7.72-7.66 (m,
2H), 7.48-7.38 / 7.36-7.30 (2m, 3H), 7.29-7.22 (m, 1H), 6.40-6.32 (m, 1H),
5.82-5.66 (m, 1H), 4.53-
4.46 / 4.30-4.23 / 3.92-3.86 / 3.37-3.22 (4m, 3H, partially concealed), 3.77-
3.65 (m, 2H), 3.65-3.59
(m, 1H), 3.55-3.42 (m, 3H), 2.90-2.81 /2.73-2.65 (2m, 1H), 2.33-2.26 / 2.26-
2.17 (2m, 2H), 2.15-
1.95 (m, 2H), 0.94-0.83 (m, 3H). Additional signals of minor rotamers were
also detected.
Example 9
5 - {2 -R7R)-11-Chloro -7-methy1-2-oxo-2,6,7,8 -tetrahydro-3H-[3]benzoxocino
[2,1-clpyridin-3 -yl] -
3 -{(2R)-1,4-dioxan-2-yll propanamido}pyridine -2 -carboxamide (mixture of two
diastereomers)
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0
IH
HG
00 I NH2
0
CI
2-[(7R)-11 -Chloro -7 -methy1-2-oxo-2,6,7,8 -tetrahydro-3H-[3]benzoxocino [2,1
-c] pyridin-3 -yl] -3 -
[(2R)-1,4-dioxan-2-yllpropanoic acid (mixture of two diastereomers) (51 mg,
0.12 mmol) and 5-
aminopyridine-2-carboxamide (24 mg, 0.18 mmol, 1.5 eq.) (prepared according to
WO 2017/005725,
example 1.3B) were reacted according to General Method 11. Yield: 50 mg (75%
of theory).
LC-MS (method 4): R1= 1.69 / 1.72 min; MS (ESIpos): m/z = 553 [M+HI
'H-NMR (500 MHz, DM50-d6): 6 [ppm] = 10.99-10.75 (m, 1H), 8.89-8.80 (m, 1H),
8.27-8.15 (m,
1H), 8.05-7.97 (m, 2H), 7.88-7.76 (m, 1H), 7.56-7.49 (m, 1H), 7.49-7.29 (m,
3H), 6.41-6.32 (m, 1H),
5.83-5.62 (m, 1H), 4.53-4.45 / 4.34-4.24 / 3.92-3.81 / 3.39-3.21 (4m, 3H,
partially concealed), 3.78-
3.57 (m, 3H), 3.55-3.41 (m, 3H), 2.90-2.81 / 2.76-2.65 (2m, 1H), 2.38-1.96 (m,
4H), 0.96-0.83 (m,
3H). Additional signals of minor rotamers were also detected.
Example 10
541(25)-2-R7R)- 11 -Chloro-7-methy1-2-oxo-2,6,7,8 -tetrahydro-3H-
[3]benzoxocino [2,1 -clpyridin-
3 -yll -3 - [(2R)-1,4 -dioxan-2 -yll propanoyllamino)pyridine -2 -carboxamide
(single stereoisomer)
*
00 0
0
HC
Nr,Nir
NH2
0
CI
,
Diastereomer separation of 45 mg of 5-{2-[(7R)-11-chloro-7-methy1-2-oxo-
2,6,7,8-tetrahydro-3H-
[3] benzoxocino [2,1 -c] pyridin-3 -yl] -3 -{(2R)-1,4 -dioxan-2 -yll
propanamidolpyridine -2 -carboxamide
(mixture of two diastereomers), Example 9 gave
single stereoisomer 1 (chiral HPLC: Rt = 3.36 min, 97% de): 14 mg,
single stereoisomer 2 (chiral HPLC: Rt = 5.43 min, 93% de): 24 mg.
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Separation method: HPLC: column: Daicel Chiralpak ID 5 um, 250 mm x 20 mm;
eluent: 25%
n-heptane / 75% ethanol; temperature: 50 C; flow rate: 15 ml/min; UV
detection: 210 nm.
Analysis method: HPLC: column: Daicel Chiralpak OX-3 3 pm, 50 mm x 4.6 mm;
eluent: 50%
n-heptane / 50% ethanol; temperature: 40 C; flow rate: 1 ml/min; UV detection:
220 nm.
.. 14 mg of single stereoisomer 1 were further purified by preparative HPLC
(reversed phase, eluent:
acetonitrile / 0.05% formic acid gradient) to give 9 mg of the title compound
Example 10.
LC-MS (method 4): R1= 1.72 min; MS (ESIpos): m/z = 553 [M+I-11+
'H-NMR (500 MHz, DM50-d6): 6 [ppm] = 10.96-10.76 (m, 1H), 8.90-8.80 (m, 1H),
8.27-8.17 (m,
1H), 8.05-7.98 (m, 2H), 7.86-7.77 (m, 1H), 7.55-7.50 (m, 1H), 7.48-7.30 (m,
3H), 6.40-6.32 (m, 1H),
5.83-5.76 / 5.68-5.62 (2m, 1H), 4.53-4.46 / 4.30-4.23 / 3.93-3.85 / 3.39-3.23
(4m, 3H, partially
concealed), 3.78-3.65 (m, 2H), 3.65-3.59 (m, 1H), 3.55-3.42 (m, 3H), 2.89-2.81
/ 2.74-2.65 (2m,
1H), 2.33-2.22 (m, 2H), 2.17-1.95 (m, 2H), 0.94-0.83 (m, 3H). Additional
signals of minor rotamers
were also detected.
Example 11
4-(1(2S,45)-24(7R)-11-Chl oro-7-methy1-2 -oxo -2,6,7, 8-tetrahydro-3H- [3]
benzoxocino [2,1-
c] pyridin-3 -yll -4-methoxypentanoyl}amino)benzamide (single stereoisomer)
C H3
C H 3
0
H 3C
N
00 N H 2
0
Ci
1,1,3,3-Tetramethylguanidine (22
178 umol, 3.0 eq.) was added under argon atmosphere at RT
to a solution of (7 R) - 11-chloro-7-methyl-7,8-dihydro-3H-[3]benzoxocino[2,1-
c]pyridin-2(6H)-one
(single stereoisomer) (17 mg, 59 umol) in 2-propanol /acetone (4:1, 0.6 m1).
The mixture was stirred
at RT for 15 min, followed by addition of 4-{[(2R,45)-2-bromo-4-
methoxypentanoyllamino}benzamide (single stereoisomer) (22 mg, 65 umol, 1.1
eq.) and of further
2-propanol / acetone (4:1, 0.6 m1). The reaction mixture was stirred at RT for
2 days and concentrated
under reduced pressure. The residue was purified by preparative HPLC (reversed
phase, eluent:
acetonitrile / water gradient). Yield: 22 mg (69% of theory).
LC-MS (method 1): Rt = 0.97 min; MS (ESIpos): m/z = 524 [M+I-11+
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'H-NMR (500 MHz, DMSO-d6): 6 [ppm] = 10.66 (br s, 1H), 7.92-7.80 (m, 4H), 7.75-
7.66 (m, 2H),
7.49-7.29 (m, 3H), 7.25 (br s, 1H), 6.40-6.31 (m, 1H), 5.87-5.76 (m, 1H), 4.54-
4.44 / 4.31-4.24 /
3.90-3.82 / 3.3-3.20 (4m, 2H, partially concealed), 3.20-3.10 (m, 3H), 2.90-
2.80 / 2.75-2.64 (2m,
2H), 2.34-2.13 / 2.10-1.96 (2m, 4H), 1.16 (d, 3H), 0.97-0.82 (m, 3H).
Additional signals of minor
rotamers were also detected.
Example 12
4-(1(2S,4R)-2- R7R)-11-Chloro-7-methy1-2 -oxo -2,6,7, 8-tetrahydro-3H- [3]
benzoxocino [2,1-
c] pyridin-3 -yll -4-cyclopropy1-4-methoxybutanoyl}amino)benzamide (single
stereoisomer)
C H 3
HC 0
N
00
NH
2
0
CI
1,1,3,3-Tetramethylguanidine (19 149 lama 3.0 eq.) was added under argon
atmosphere at RT
to a solution of (7 R) - 11-chloro-7-methyl-7,8-dihydro-3H-[3]benzoxocino[2,1-
c]pyridin-2(6H)-one
(single stereoisomer) (14 mg, 50 mop in 2-propanol / acetone (4:1, 0.5 m1).
The mixture was stirred
at RT for 15 min, followed by addition of 4-{[(2R,4R)-2-bromo-4-cyclopropy1-4-
methoxybutanoyll-
aminolbenzamide (single stereoisomer) (19 mg, 55 lama 1.1 eq.) and of further
2-propanol / acetone
(4:1, 0.5 m1). The reaction mixture was stirred at RT overnight and
concentrated under reduced
pressure. The residue was purified by preparative HPLC (reversed phase,
eluent: acetonitrile / water
gradient). Yield: 16 mg (59% of theory).
LC-MS (method 4): R1= 1.95 min; MS (ESIpos): m/z = 550 [M+H1+
'H-NMR (600 MHz, DM50-d6): 6 [ppm] = 10.69-10.54 (m, 1H), 7.90-7.74 (m, 4H),
7.73-7.66 (m,
2H), 7.48-7.28 (m, 3H), 7.22 (br s, 1H), 6.38-6.30 (m, 1H), 5.88-5.75 (m, 1H),
4.54-4.45 /4.33-4.26
/3.36-3.76 / 3.3-3.26 (4m, 2H, partially concealed), 3.23 / 3.21 (2s, 3H),
2.89-2.81 /2.79-2.67 (2m,
1H), 2.45-2.21 (m, 4H), 2.11-1.97 (m, 1H), 0.96-0.77 (m, 4H), 0.63-0.51 (m,
1H), 0.45-0.33 (m, 2H),
0.10-0.0 (m, 1H). Additional signals of minor rotamers were also detected.
Example 13
4- {2 -[(7R)-11 -Chloro -7-methy1-2-oxo-2,6,7,8 -tetrahydro-3H-[3]benzoxocino
[2,1 -clpyridin-3 -yl] -
4-(cyclopropyloxy)butanamido}benzamide (mixture of two diastereomers)
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,j\
0 \11
H 3C
N"(
00 N H 2
0
"CI
4-Aminobenzamide (3 mg, 25 ma 1.2 eq.), pyridine (2 [11, 25 ma 1.2 eq.) and
T3P (19 pi, 50%
solution in ethyl acetate, 31 ma 1.5 eq.) were added under argon atmosphere
at RT to a solution
of 2-[(7R)-11 -chloro-7-methyl-2 -oxo -2,6,7,8-tetrahydro-3H- [3] benzoxocino
[2,1-c] pyridin-3 -yll -4-
(cyclo-propyloxy)butanoic acid (mixture of two diastereomers) (9 mg, 21 [tmol)
in tetrahydrofuran
(1 m1). The reaction mixture was stirred at RT for 1.5 h before water was
added. After phase
separation, the aqueous phase was extracted with ethyl acetate. The combined
organic phases were
dried over anhydrous sodium sulfate, filtered and concentrated under reduced
pressure. The residue
was purified by preparative HPLC (reversed phase, eluent: acetonitrile / water
gradient). Yield: 5 mg
(44% of theory).
LC-MS (method 4): R1= 1.90 min; MS (ESIpos): m/z = 536 [M+H1+
'H-NMR (500 MHz, DM50-d6): 6 [ppm] = 10.73-10.61 (m, 1H), 7.91-7.76 (m, 4H),
7.73-7.64 (m,
2H), 7.49-7.21 (m, 4H), 6.41-6.31 (m, 1H), 5.78-5.69 (m, 1H), 4.54-4.45 / 4.32-
4.26 / 3.91-3.84 /
3.58-3.06 (4m, 4H, partially concealed), 2.91-2.82 / 2.77-2.69 (2m, 1H), 2.4-
2.26 (m, 4H, partially
concealed), 2.11-1.97 (m, 1H), 0.97-0.85 (m, 3H), 0.45-0.28 (m, 4H).
Additional signals of minor
rotamers were also detected.
Example 14
4-{ [2 - [(7R)-11-Chloro-7-methy1-2 -oxo -2,6,7,8-tetrahydro-3H-[3]benzoxocino
[2,1-c] pyridin-3 -yl] -
4-(cyclobutyloxy)butanoyl] amino}benzamide (mixture of two diastereomers)
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a/0
0
H 3C
N
0 1411 N H 2
0
"CI
4-Aminobenzamide (42 mg, 0.31 mmol, 1.5 eq.) and /V,N-diisopropylethylamine
(107 [11, 0.61 mmol,
3.0 eq.) were added at RT to a solution of 2-[(7R)-11-chloro-7-methy1-2-oxo-
2,6,7,8-tetrahydro-3H-
[3]benzoxocino[2,1-clpyridin-3-y1]-4-(cyclobutyloxy)butanoic acid (mixture of
two diastereomers)
(106 mg, 83% purity, 0.20 mmol) in /V,N-dimethylformamide (1 m1). A solution
of HATU (116 mg,
0.31 mmol, 1.5 eq.) in /V,N-dimethylformamide (2 ml) was added dropwise and
the reaction mixture
stirred at RT for 2 days. Additional HATU (39 mg, 0.10 mmol, 0.5 eq.) and /V,N-
diisopropylethylamine (36 [11, 0.20 mmol, 1.0 eq.) were added. The reaction
mixture was stirred for
3 h before water was added. After addition of ethyl acetate and phase
separation, the aqueous phase
was extracted ethyl acetate. The combined organic phases were dried over
anhydrous sodium sulfate,
filtered and concentrated under reduced pressure. The residue was purified by
preparative HPLC
(reversed phase, eluent: acetonitrile / water gradient). Yield: 31 mg (27% of
theory).
LC-MS (method 4): R1= 1.99 min; MS (ESIpos): m/z = 550 [M+H1+
'H-NMR (600 MHz, DM50-d6): 6 [ppm] = 10.75-10.60 (m, 1H), 7.93-7.77 (m, 4H),
7.74-7.65 (m,
2H), 7.49-7.20 (m, 4H), 6.41-6.31 (m, 1H), 5.81-5.71 (m, 1H), 4.53-4.44 / 4.32-
4.25 / 3.89-3.78 /
3.39-3.17 (4m, 5H, partially concealed), 2.89-2.81 / 2.75-2.68 (2m, 1H), 2.41-
2.24 (m, 3H), 2.12-
1.96 (m, 3H), 1.30-1.49 / 1.47-1.35 (2m, 4H), 0.96-0.83 (m, 3H). Additional
signals of minor
rotamers were also detected.
Example 15
4-{ R25)-2 -(11-Chloro -12 -fluoro-7-methyl-2-oxo-2,6,7,8-tetrahydro-3H- [3]
benzoxocino [2,1-
c] pyridin-3 -yl)butanoyll amino -2 -fluorobenzamide (mixture of two diaste re
omers)
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C H 3
H
H 3C
N
0 N H 2
100 F 0
CI
General Method 7 was carried out with 11-chloro-12-fluoro-7-methy1-7,8-dihydro-
3H-
[3]benzoxocino[2,1-clpyridin-2(6H)-one (racemate) (30.0 mg, 92% purity, 94.0
umol, 1.0 eq.), 4-
(2R)-2-bromobutanoyllaminol -2-fluorobenzamide (single stereoisomer) (42.7 mg,
141 umol, 1.5
eq.) and 1,1,3,3-tetramethylguanidine (35 IA, 280 umol, 3.0 eq.) in a mixture
of 2-propanol / acetone
(4:1, 1.0 ml) including the following variations of the procedure: The crude
mixture was purified by
preparative HPLC (reversed phase, eluent: water with 0.05% formic acid /
acetonitrile 90:10 to 5:95).
Yield: 35.0 mg (72% of theory).
LC-MS (method 3): R1= 3.09 / 3.16 min; MS (ESIpos): m/z = 516 11M+I-11+
Example 16
4-(1(25)-2-R7R)- 11 -Chloro-12-fluoro-7 -methy1-2 -oxo -2,6,7,8-tetrahydro-3H-
[3] benzoxocino 112, 1 -
c] pyridin-3 -yll butanoyl amino)-2-fluorobenzamide (single stereoisomer)
C H 3
0 H
H 3C
N
00 N H 2
F 0
CI
Diastereomer separation of 30 mg of 4-{ R25)-2-(11-chloro-12-fluoro-7-methy1-2-
oxo-2,6,7,8-
tetrahydro-3H-[3]benzoxocino 112, 1 -c] pyridin-3 -yl)butanoyll amino} -2 -
fluorobenzamide (mixture of
two diastereomers), Example 15 gave
single stereoisomer 1 (the title compound 16) (chiral HPLC: R1= 11.5 min, >98%
de): 11.0 mg (23%
of theory),
single stereoisomer 2 (chiral HPLC: R1 = 15.3 min): 11.0 mg.
Separation method: HPLC: column: Daicel Chiralpak IA 5 um, 250 mm x 20 mm;
eluent: ethanol;
temperature: 20 C; flow rate: 7 ml/min; UV detection: 240 nm.
Analysis method: HPLC: column: Daicel Chiralpak IA 5 um, 250 mm x 4.6 mm;
eluent: ethanol;
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temperature: 20 C; flow rate: 0.4 ml/min; UV detection: 240 nm.
LC-MS (method 3): R1= 3.36 min; MS (ESIpos): m/z = 516 [M+H1+
1H-NMR (400 MHz, DM50-d6): 6 [ppm] = 10.89 /10.86 (2s, 1H), 7.89 (s, 1H), 7.73-
7.57 (m, 3H),
7.56-7.51 (m, 2H), 7.43-7.37 (m, 1H), 7.30 / 7.22 (2d, 1H), 6.49 / 6.45 (2d,
1H), 5.66-5.50 (m, 1H),
4.50 (dd, 1H), 3.33-3.27 (m, 1H), 2.77 (d, 1H), 2.32-2.24 (m, 1H), 2.31-1.99
(m, 3H), 0.96-0.82 (m,
6H). Additional signals of minor rotamers were also detected.
Example 17
4-(1(25)-2411-Chloro -12 -fluoro-7-methyl-2-oxo-2,6,7,8-tetrahydro-3H- [3]
benzoxocino [2,1-
c] pyridin-3 -y1)-3 - [(25)-tetrahydro-2H-pyran-2-yll propanoyllamino)-2-
fluorobenzamide (mixture
of two diastereomers)
0
0
H 3C
N
00 N H 2
= F 0
CI
General Method 7 was carried out with 11-chloro-12-fluoro-7-methy1-7,8-dihydro-
3H-
[3]benzoxocino[2,1-clpyridin-2(6H)-one (racemate) (50.0 mg, 92% purity, 157
umol, 1.0 eq.), 4-
( (2R)-2 -bromo -3 -[(25)-tetrahydro-2H-pyran-2-yll propanoyl}amino)-2 -
fluorobenzamide (single
stereoisomer) (87.7 mg, 235 umol, 1.5 eq.) and 1,1,3,3-tetramethylguanidine
(59 IA, 470 umol, 3.0
eq.) in a mixture of 2-propanol / acetone (4:1, 1.5 ml) including the
following variations of the
procedure: The crude mixture was purified by preparative HPLC (reversed phase,
eluent: water with
0.05% formic acid / acetonitrile 90:10 to 5:95). Yield: 65.0 mg (71% of
theory).
LC-MS (method 4): Rt = 2.00 min; MS (ESIpos): m/z = 586 [M+H1+
Example 18
4-(1(25)-2-R7R)- 11 -Chloro-12-fluoro-7-methy1-2 -oxo -2,6,7,8-tetrahydro-3H-
[3]benzoxocino [2,1-
c] pyridin-3 -yll -3 - [(25)-tetrahydro-2H-pyran-2-yll propanoyllamino)-2-
fluorobenzamide (single
stereoisomer)
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0 Y, 1
H 3C
N,c
0 10
NH
2
F 0
CI
Diastereomer separation of 60 mg of 4-({(25)-2-(11-chloro-12-fluoro-7-methyl-2-
oxo-2,6,7,8-
tetrahydro-3H-[3]benzoxocino [2, 1-c] pyridin-3 -y1)-3 -[(25)-tetrahydro-2H-
pyran-2 -
yllpropanoyl}amino)-2-fluorobenzamide (mixture of two diastereomers), Example
17 gave
single stereoisomer 1 (the title compound 18) (chiral HPLC: R1= 36.3 min, >98%
de): 12.0 mg (13%
of theory),
single stereoisomer 2 (chiral HPLC: Rt = 48.6 min): 15.0 mg.
Separation method: HPLC: column: two coupled Daicel Chiralpak OD-H 5 pm, 250
mm x 20 mm;
eluent: 90% n-heptane / 5% 2-propanol / 5% ethanol; temperature: 20 C; flow
rate: 8.0 ml/min; UV
detection: 240 nm.
Analysis method: HPLC: column: two coupled Daicel Chiralpak OD-H 5 pm, 250 mm
x 4.6 mm;
eluent: 90% n-heptane / 5% methanol / 5% ethanol; temperature: 20 C; flow
rate: 1.0 ml/min; UV
detection: 240 nm.
LC-MS (method 3): Rt = 3.71 min; MS (ESIpos): m/z = 586 [M-411+
1H-NMR (400 MHz, DM50-d6): 6 [ppm] = 10.84 / 10.75 (2s, 1H), 7.90 / 7.87 (2s,
1H), 7.74-7.57
(m, 3H), 7.56-7.50 (m, 2H), 7.47-7.42 (m, 1H), 7.29 (d, 1H), 6.49-6.41 (m,
1H), 5.79 / 5.72 (2t, 1H),
4.49 (dd, 1H), 3.85-3.79 (m, 1H), 3.29-3.21 (m, 2H), 2.75 (d, 1H), 2.33-2.21
(m, 2H), 2.20-1.95 (m,
2H), 1.81-1.70 (m, 1H), 1.61 (d, 1H), 1.48-1.39 (m, 3H), 1.32-1.21 (m, 2H),
0.90 / 0.84 (2t, 3H).
Additional signals of minor rotamers were also detected.
Example 19
4-(1(25)-2411-Chloro -12 -fluoro-7-methyl-2-oxo-2,6,7,8-tetrahydro-3H- [3]
benzoxocino [2,1-
c] pyridin-3 -y1)-3 - [(25)-tetrahydro-2H-pyran-2-yll
propanoyllamino)benzamide (mixture of two
diastereomers)
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0
0
H 3C
N
00
N H 2
= F 0
CI
General Method 7 was carried out with 11-chloro-12-fluoro-7-methy1-7,8-dihydro-
3H-
[3]benzoxocino[2,1-clpyridin-2(6H)-one (racemate) (50.0 mg, 92% purity, 157
umol, 1.0 eq.), 4-
( (2R)-2 -bromo -3 -[(25)-tetrahydro-2H-pyran-2-yll propanoyl}amino)benzamide
(single
stereoisomer) (83.4 mg, 235 umol, 1.5 eq.) and 1,1,3,3-tetramethylguanidine
(59 IA, 470 umol, 3.0
eq.) in a mixture of 2-propanol / acetone (4:1, 1.5 ml) including the
following variations of the
procedure: The crude mixture was purified by preparative HPLC (reversed phase,
eluent: water with
0.05% formic acid! acetonitrile 90:10 to 5:95). Yield: 57 mg (64% of theory).
LC-MS (method 3): Rt = 3.36! 3.42 min; MS (ESIpos): m/z = 568 [M-411+
Example 20
4-( (25)-2 -{(7R)- 11 -Chloro-12-fluoro-7-methy1-2 -oxo -2,6,7,8-tetrahydro-3H-
[3]benzoxocino [2,1-
c] pyridin-3 -yll -3 - [(25)-tetrahydro-2H-pyran-2-yll
propanoyllamino)benzamide (single
stereoisomer)
0
cLF'r
H 3C
N
00
N H 2
F 0
CI
Diastereomer separation of 55 mg of 4-({(25)-2-(11-chloro-12-fluoro-7-methyl-2-
oxo-2,6,7,8-
tetrahydro-3H-[3]benzoxocino [2, 1 -c] pyridin-3 -y1)-3 -[(25)-tetrahydro-2H-
pyran-2-
yllpropanoyl}amino)-benzamide (mixture of two diastereomers), Example 19 gave
single stereoisomer 1 (the title compound 20) (chiral HPLC: R1= 43.7 min, >95%
de): 11.0 mg (11%
of theory),
single stereoisomer 2 (chiral HPLC: Rt = 62.6 min): 13.0 mg.
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Separation method: HPLC: column: two coupled Daicel Chiralpak OD-H 5 pm, 250
mm x 20 mm;
eluent: 90% n-heptane / 5% 2-propanol / 5% ethanol; temperature: 20 C; flow
rate: 15 ml/min; UV
detection: 240 nm.
Analysis method: HPLC: column: two coupled Daicel Chiralpak OD-H 5 pm, 250 mm
x 4.6 mm;
eluent: 90% n-heptane / 5% 2-propanol / 5% ethanol; temperature: 20 C; flow
rate: 1.0 ml/min; UV
detection: 240 nm.
LC-MS (method 3): Rt = 3.50 min; MS (ESIpos): m/z = 568 [M-411+
1H-NMR (400 MHz, DM50-c/6): 6 [ppm] = 10.68 /10.61 (2s, 1H), 7.93-7.80 (m,
4H), 7.74-7.67 (m,
2H), 7.62-7.57 (m, 1H), 7.33-7.19 (m, 2H), 6.49-6.41 (m, 1H), 5.82 / 5.77 (2t,
1H), 4.51-4.47 (m,
1H), 3.85-3.79 (m, 1H), 3.29-3.21 (m, 2H), 2.75 (d, 1H), 2.33-2.24 (m, 2H),
2.25-2.17 (m, 1H), 2.14-
1.99 (m, 1H), 1.81-1.70 (m, 1H), 1.62 (d, 1H), 1.49-1.35 (m, 3H), 1.35-1.14
(m, 2H), 0.91 / 0.84 (2t,
3H). Additional signals of minor rotamers were also detected.
Example 21
4-{ [(2S)-2 -(11-Chloro -7 -ethy1-2-oxo-2,6,7,8 -tetrahydro-3H- [3]
benzoxocino [2,1-c] pyridin-3 -
yl)butanoyllamino}-2-fluorobenzamide (mixture of two diastereomers)
C H 3
0 -I
el
H 3 C N 0
NH 2
= 0
C I
General Method 7 was carried out with 11-chloro-7-ethy1-7,8-dihydro-3H-
[3]benzoxocinop,1-
clpyridin-2(6H)-one (racemate) (25.0 mg, 78% purity, 67.3 umol, 1.0 eq.), 4-
{[(2R)-2-
bromobutanoyllamino}-2-fluorobenzamide (single stereoisomer) (40.8 mg, 135
umol, 2.0 eq.) and
1,1,3,3-tetramethylguanidine (25 jil, 200 umol, 3.0 eq.) in a mixture of 2-
propanol / acetone (4:1,
0.9 ml) including the following variations of the procedure: The crude mixture
was purified by
preparative HPLC (reversed phase, eluent: water with 0.05% formic acid /
acetonitrile 90:10 to 5:95).
Yield: 42.0 mg (quantitative of theory).
LC-MS (method 1): R1 = 1.09 min; MS (ESIpos): m/z = 512 [M-411+
Example 22
4-(1(25)-2-R7R)- 11 -Chloro-7-ethy1-2 -oxo -2,6,7, 8-tetrahydro-3H- [3]
benzoxocino [2,1-clpyridin-3-
yllbutanoyl}amino)-2-fluorobenzamide (single stereoisomer)
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C H 3
0 N(
o.,(1-1\11
N
H 3C
N H 2
= 0
CI
Diastereomer separation of 37 mg of 4-{ [(2S)-2-(11-chloro-7-ethy1-2-oxo-
2,6,7,8-tetrahydro-3H-
[3] benzoxocino [2,1-c] pyridin-3 -yl)butanoyll amino -2-fluorobenzamide
(mixture of two
diastereomers), Example 21 gave
single stereoisomer 1 (the title compound 22) (chiral HPLC: R1 = 1.26 min, 99%
de): 10.8 mg (31%
of theory),
single stereoisomer 2 (chiral HPLC: R1 = 2.70 min): 10.2 mg.
Separation method: HPLC: column: Daicel Chiralpak ID 5 [tm, 250 mm x 20 mm;
eluent: 50%
n-heptane / 50% ethanol; temperature: 40 C; flow rate: 20 ml/min; UV
detection: 220 nm.
Analysis method: HPLC: column: Daicel Chiralpak ID-3 3 [tm, 50 mm x 4.6 mm;
eluent: 50%
n-heptane / 50% ethanol; flow rate: 1.0 ml/min; UV detection: 220 nm.
LC-MS (method 3): R1= 3.57 min; MS (ESIpos): m/z = 512 [M+I-11+
1H-NMR (400 MHz, DM50-d6): 6 [ppm] = 10.87 / 10.84 (2s, 1H), 7.78 (s, 1H),
7.73-7.62 (m, 2H),
7.58-7.50 (m, 2H), 7.49-7.44 (m, 1H), 7.42-7.32 (m, 3H), 6.39 / 6.34 (2s, 1H),
5.61-5.52 (m, 1H),
4.58-4.47 (m, 1H), 3.45-3.35 (m, 1H), 2.77 (br d, 1H), 2.28-2.02 (m, 3H), 1.86-
1.68 (m, 1H), 1.36-
1.16 (m, 2H), 0.99 (t, 3H), 0.89 (t, 3H). Additional signals of minor rotamers
were also detected.
Example 23
4-{ R25)-2 -(11-Chloro -6,7-dimethy1-2 -oxo-2,6,7,8-tetrahydro-3H- [3]
benzoxocino [2,1-c] pyridin-3 -
yl)butanoyllamino}-2-fluorobenzamide (mixture of stereoisomers)
HC C H3
0
H 3C N
o N H2
0
C
I
General Method 7 was carried out with 11-chloro-6,7-dimethy1-7,8-dihydro-3H-
[3]benzoxocino [2,1-
clpyridin-2(6H)-one (mixture of stereoisomers) (65.0 mg, 224 lama 1.0 eq.), 4-
{[(2R)-2-
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bromobutanoyllamino}-2-fluorobenzamide (single stereoisomer) (68.0 mg, 224
umol, 1.0 eq.) and
1,1,3,3-tetramethylguanidine (84 jil, 670 umol, 3.0 eq.) in a mixture of 2-
propanol / acetone (4:1,
3.75 ml) including the following variations of the procedure: After stirring
overnight, additional
amounts of 4-{(2R)-2-bromobutanoyllamino}-2-fluorobenzamide (single
stereoisomer) (20.4 mg,
67.2 umol, 0.3 eq.) and 1,1,3,3-tetramethylguanidine (21 jil, 168 umol, 0.75
eq.) were added and
stirring was continued at RT for 3 h. The crude mixture was purified by
preparative HPLC (reversed
phase, eluent: water with 0.05% formic acid / acetonitrile 80:20 to 5:95).
Yield: 55.0 mg (48% of
theory).
LC-MS (method 1): R1= 1.03 / 1.05 min; MS (ESIpos): m/z = 512 [M+I-11+
Example 24
4-( (25)-2 -R6R,7 R)-11-Chloro-6,7-dimethy1-2 -oxo -2,6,7,8-tetrahydro-3H-[3]
benzoxocino [2,1-
c] pyridin-3 -yll butanoyl amino)-2-fluorobenzamide (single stereoisomer)
C H 3
H 3C
0 1\11
H 3C N
0 N H
0
CI
Stereoisomer separation of 50 mg of 4-{ R25)-2-(11-chloro-6,7-dimethy1-2-oxo-
2,6,7,8-tetrahydro-
3H-[3]benzoxocino [2,1 -clpyridin-3 -yl)butanoyll amino} -2 -fluorobenzamide
(mixture of
stereoisomers), Example 23 gave
single stereoisomer 1 (the title compound 24) (chiral HPLC: Rt = 5.25 min, 99%
de): 5.60 mg (5%
of theory),
single stereoisomer 2 (chiral HPLC: Rt = 6.01 min): 1.70 mg,
single stereoisomer 3 (chiral HPLC: Rt = 7.18 min): 17.3 mg,
single stereoisomer 4 (chiral HPLC: Rt = 8.65 min): 18.3 mg.
Separation method: HPLC: column: Daicel Chiralpak IF 5 um, 250 mm x 20 mm;
eluent: 60%
n-heptane / 40% ethanol; temperature: 40 C; flow rate: 15 ml/min; UV
detection: 220 nm.
Analysis method: HPLC: column: Daicel Chiralpak IF 5 um, 250 mm x 4.6 mm;
eluent: 60%
iso-hexane / 40% ethanol; flow rate: 1.0 ml/min; UV detection: 220 nm.
LC-MS (method 1): Rt = 1.07 min; MS (ESIpos): m/z = 512 [M+I-11+
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1H-NMR (500 MHz, DMSO-d6): 6 [ppm] = 10.82 (s, 1H), 7.77 (s, 1H), 7.73-7.60
(m, 2H), 7.57-7.47
(m, 2H), 7.46-7.38 (m, 3H), 7.37-7.30 (m, 1H), 6.40 / 6.35 (2s, 1H), 5.59 (dd,
1H), 3.58-3.45 (m,
1H), 2.70-2.59 (m, 1H), 2.44-2.33 (m, 1H), 2.23-2.03 (m, 2H), 1.78-1.62 (m,
1H), 1.38 /1.32 (2d,
3H), 1.01 (d, 3H), 0.89/ 0.77 (2t, 3H). Additional signals of minor rotamers
were also detected.
Example 25
4-(1(25)-2411-Chloro -6,7-dimethy1-2 -oxo-2,6,7,8-tetrahydro-3H- [3]
benzoxocino [2,1-c] pyridin-3 -
y1)-3 -[(25)-tetrahydro -2H-pyran-2-yll propanoyl}amino)benzamide (mixture of
ste reoi some rs)
0,-
H 3C
0 11.16
H 3C
N'(
00 .1
NH
2
0
CI
General Method 7 was carried out with 11-chloro-6,7-dimethy1-7,8-dihydro-3H-
[3]benzoxocino [2,1-
c]pyridin-2(6H)-one (mixture of stereoisomers) (25.0 mg, 86.3 lama 1.0 eq.), 4-
({(2R)-2-bromo-3-
[(25)-tetrahydro-2H-pyran-2-yllpropanoyllamino)benzamide (single stereoisomer)
(46.0 mg, 129
lama 1.5 eq.) and 1,1,3,3-tetramethylguanidine (320, 260 lama 3.0 eq.) in a
mixture of 2-propanol
/ acetone (4:1, 0.5 ml) and the crude mixture was purified by preparative HPLC
(reversed phase,
eluent: water with 0.05% formic acid / acetonitrile 80:20 to 5:95). Yield:
38.7 mg (79% of theory).
LC-MS (method 4): Rt = 2.02 min; MS (ESIpos): m/z = 564 [M+H1+
Example 26
4-(1(25)-24(6R,7R)-11-Chloro-6,7-dimethyl-2 -oxo -2,6,7,8-tetrahydro-3H- [3]
benzoxocino [2,1-
c] pyridin-3 -yll -3 - [(25)-tetrahydro-2H-pyran-2-yll
propanoyllamino)benzamide (single
stereoisomer)
H 3C
0
H 3C
N
00 .1 N H 2
0
CI
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Stereoisomer separation of 35 mg of 4-({(25)-2-(11-chloro-6,7-dimethyl-2-oxo-
2,6,7,8-tetrahydro-
3H-[3]benzoxocino [2,1 -c] pyridin-3 -y1)-3 - [(25)-tetrahydro-2H-pyran-2-
yllpropanoyllamino)benzamide (mixture of stereoisomers), Example 25 gave
single stereoisomer 1 (the title compound 26) (chiral HPLC: Rt = 6.41 min, 99%
de): 6.0 mg (12%
of theory),
single stereoisomer 2 (chiral HPLC: Rt = 9.80 min): 7.0 mg,
mixture of stereoisomers (chiral HPLC: R1= 6.43 /8.10 min): 3.0 mg.
Separation method: HPLC: column: Daicel Chiralpak IE 5 um, 250 mm x 20 mm;
eluent: 50%
n-heptane / 50% ethanol; temperature: 40 C; flow rate: 15 ml/min; UV
detection: 220 nm.
Analysis method: HPLC: column: Daicel Chiralpak IE 5 um, 250 mm x 4.6 mm;
eluent: 50%
iso-hexane / 50% ethanol; flow rate: 1.0 ml/min; UV detection: 220 nm.
LC-MS (method 1): R1= 1.07 min; MS (ESIpos): m/z = 564 [M-411+
1H-NMR (500 MHz, CDC13): 6 [ppm] = 9.80 (s, 1H), 7.84-7.75 (m, 2H), 7.74-7.65
(m, 2H), 7.36-
7.31 (m, 2H), 7.23-7.17 (m, 2H), 6.56 (s, 1H), 5.93 (dd, 1H), 6.20-5.57 (m,
2H), 4.09-3.94 (m, 1H),
3.60-3.50 (m, 1H), 3.45-3.36 (m, 2H), 2.63-2.46 (m, 3H), 1.97-1.47 (m, 8H),
1.40 (d, 3H), 1.00 (d,
3H). Additional signals of minor rotamers were also detected.
Example 27
4-{ R25)-2 -(cis-6-Chloro-3 -oxo-3 ,9,9a, 10,11, lla-hexahydro-2H-
cyclobuta[4,51 [3]benzoxocino [2,1-
clpyridin-2-yl)butanoyll amino -2 -fluorobenzamide (mixture of two
diastereomers)
C H 3
=0 N,rN F
00I 0
N H 2
C
I
1,1,3,3-Tetramethylguanidine (46
0.37 mmol, 3.0 eq.) was added under argon atmosphere at RT
to
a solution of cis-6-chloro-9a,10, 11,11a-tetrahydro-2H-cyc10but44,5] [3]
benzoxocino [2,1 -
c] pyridin-3 (9H)-one (racemate) (35 mg, 0.12 mmol) in 2-propanol / acetone
(4:1, 3.75 m1). The
mixture was stirred at RT for 15 min, followed by addition of 4-{ [(2R)-2-
bromobutanoyllamino}-2-
fluorobenzamide (single stereoisomer) (55 mg, 0.18 mmol, 1.5 eq.). The
reaction mixture was stirred
at RT overnight and concentrated under reduced pressure. The residue was
purified by preparative
HPLC (reversed phase, eluent: acetonitrile / water gradient). Yield: 43 mg
(69% of theory).
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LC-MS (method 4): R1= 1.93 min; MS (ESIpos): m/z = 510 [M+I-11+
Example 28
4-( { (25)-2 -{(9aR,11aR)-6-Chloro-3 -oxo-3 ,9,9a,10,11,11a-hexahydro-2H-
cyc10but44,5] [3] benz-
oxocino [2,1-c] pyridin-2 -yll butanoyl}amino)-2-fluorobenzamide (single
stereoisomer)
C H 3
=0 NrN F
o
= N H 2
CI
Diastereomer and atropisomer separation of 32 mg of 4-{ [(25)-2-(cis-6-chloro-
3-oxo-
,9,9a, 10, 11,11a-hexahydro -2H-cyc10but44,5] [3]benzoxocino [2,1 -c] pyridin-
2-
yl)butanoyll amino -2-fluorobenzamide (mixture of two diastereomers), Example
27 gave
single stereoisomer 1 (the title compound Example 28), (chiral HPLC: R1 = 2.81
min, >99% de): 5
mg,
single stereoisomer 2 (chiral HPLC: R1 = 3.37 min): 4 mg,
single stereoisomer 3 (chiral HPLC: R1 = 4.57min): 5 mg,
single stereoisomer 4 (chiral HPLC: R1 = 5.35 min): 3 mg.
Separation method: HPLC: column: Daicel Chiralpak ID 5 pm, 250 mm x 20 mm;
eluent: 50%
n-heptane / 50% ethanol; temperature: 30 C; flow rate: 20 ml/min; UV
detection: 265 nm.
Analysis method: HPLC: column: Daicel Chiralpak ID 5 pm, 250 mm x 4.6 mm;
eluent: 50%
n-heptane / 50% ethanol; temperature: 50 C; flow rate: 1 ml/min; UV detection:
220 nm.
LC-MS (method 4): R1= 1.91 min; MS (ESIpos): m/z = 510 [M+I-11+
1H-NMR (500 MHz, DM50-d6): 6 [ppm] = 10.93-10.74 (m, 1H), 7.70 (br t, 1H),
7.64 (br d, 1H),
7.53 (br d, 2H), 7.48-7.38 (m, 4H), 7.36 (br s, 1H), 6.39 (s, 1H), 5.66-5.53
(m, 1H), 4.65-4.56 (m,
1H), 2.78-2.65 (m, 2H), 2.65-2.54 (m, 2H), 2.21-1.99 (m, 3H), 1.82-1.69 (m,
1H), 1.40-1.20 (m, 1H),
0.92-0.83 (m, 3H).
Example 29
4-(1(25)-2 -kis-6-Chloro-3 -oxo-3 ,9,9a, 10,11, lla-hexahydro-2H-
cyclobuta[4,51 [3]benzoxocino [2,1-
clpyridin-2-yllbutanoyllamino)-benzamide (mixture of two diastereomers)
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C H 3
= 0 N.rN
00 0
= N H 2
Ci
1,1,3,3-Tetramethylguanidine (66 [11, 0.53 mmol, 3.0 eq.) was added under
argon atmosphere at RT
to a solution of cis-6-chloro-9a,10, 11,11a-tetrahydro-2H-cyc10but44,5]
[3] benzoxocino [2,1 -
c] pyridin-3 (911)-one (racemate) (51 mg, 0.18 mmol) in 2-propanol / acetone
(4:1, 5.5 m1). The
mixture was stirred at RT for 15 min, followed by addition of 4-{[(2R)-2-
bromobutanoyllamino}benzamide (single stereoisomer) (76 mg, 0.27 mmol, 1.5
eq.). The reaction
mixture was stirred at RT overnight and concentrated under reduced pressure.
The residue was
purified by preparative HPLC (reversed phase, eluent: acetonitrile / water
gradient). Yield: 39 mg
(43% of theory).
LC-MS (method 4): R1= 1.81 min; MS (ESIpos): m/z = 492 [M+141+
Example 30
4-( { (25)-2 -{(9aR,11aR)-6-Chloro-3 -oxo-3 ,9,9a, 10,11,11a-hexahydro-2H-
cyclobuta[4,5] [3] benz -
oxocino [2,1 -c] pyridin-2 -yll butanoyl}amino)benzamide (single stereoisomer)
C H3
00I 0
N H 2
Ci
Diastereomer and atropisomer separation of 35 mg of 4-({(25)-2-{cis-6-chloro-3-
oxo-
3 ,9,9a, 10, 11,11a-hexahydro -2H-cyc10but44,5] [3] benzoxocino [2,1 -c]
pyridin-2-
yllbutanoyl}amino)benzamide (mixture of two diastereomers), Example 29 gave
single stereoisomer 1 (the title compound Example 30) (chiral HPLC: Rt = 7.09
min, >99% de): 5
mg,
single stereoisomer 4 (chiral HPLC: Rt = 8.30 min): 3 mg,
single stereoisomer 3 (chiral HPLC: Rt = 10.81min): 1 mg,
single stereoisomer 2 (chiral HPLC: Rt = 12.50 min): 9 mg.
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Separation method: HPLC: column: Daicel Chiralpak IE 5 lam, 250 mm x 20 mm;
eluent: 50%
n-heptane / 50% ethanol; temperature: 60 C; flow rate: 15 ml/min; UV
detection: 220 nm.
Analysis method: HPLC: column: Daicel Chiralpak IE 5 lam, 250 mm x 4.6 mm;
eluent: 50%
n-heptane / 50% ethanol; temperature: 50 C; flow rate: 1 ml/min; UV detection:
220 nm.
LC-MS (method 4): R1= 1.83 min; MS (ESIpos): m/z = 492 [M+F11+
1H-NMR (600 MHz, DM50-d6): 6 [ppm] = 10.72-10.63 (m, 1H), 7.94-7.81 (m, 3H),
7.67 (d, 2H),
7.49-7.33 (m, 4H), 7.26 (br s, 1H), 6.38 (s, 1H), 5.70-5.58 (m, 1H), 4.64-4.57
(m, 1H), 2.79-2.66 (m,
2H), 2.63-2.56 (m, 2H), 2.21-2.01 (m, 3H), 1.81-1.65 (m, 1H), 1.37-1.21 (m,
1H), 0.89 (t, 3H).
Example 31
4-( { (25)-2 -[(9aR,11aR)-6-Chloro-3 -oxo-3 ,9,9a, 10,11,11a-hexahydro-2H-
cyclobuta[4,5] [3]benz-
oxocino [2,1-c] pyridin-2 -yll -3 -[(25)-tetrahydro-2H-pyran-2 -yll
propanoyl}amino)benzamide (single
stereoisomer)
0
NX'
+ 0
0 VI 0
0
= N H 2
CI
1,1,3,3-Tetramethylguanidine (33 [11, 0.26 mmol, 3.0 eq.) was added under
argon atmosphere at RT
to a solution of .. (9aR,
11aR)-6-chloro-9a, 10,11, lla-tetrahydro-2H-
cyclobuta[4,51[3]benzoxocino[2,1-clpyridin-3(91-1)-one (single stereoisomer)
(25 mg, 0.09 mmol) in
2-propanol / acetone (4:1, 2.7 m1). The mixture was stirred at RT for 15 min,
followed by addition
of 4-({ (2R)-2-bromo -3 - [(25)-tetrahydro-2H-pyran-2-yll
propanoyllamino)benzamide (single
stereoisomer) (46 mg, 0.13 mmol, 1.5 eq.). The reaction mixture was stirred at
RT overnight, at 40 C
for further 22 h and concentrated under reduced pressure. The residue was
purified by preparative
HPLC (reversed phase, eluent: acetonitrile / water gradient). Yield: 30 mg
(61% of theory).
LC-MS (method 4): R1= 1.99 min; MS (ESIpos): m/z = 562 [M+F11+
'H-NMR (600 MHz, DM50-d6): 6 [ppm] = 10.75-10.57 (m, 1H), 7.92-7.78 (m, 3H),
7.70 (d, 2H),
7.54-7.30 (m, 4H), 7.26 (br s, 1H), 6.37 (s, 1H), 5.81 (br t, 1H), 4.60 (q,
1H), 3.85 (br d, 1H), 3.3-
3.16 (m, 2H, partially concealed), 2.82-2.63 (m, 2H), 2.63-2.5 (m, 2H,
partially concealed), 2.30-
2.01 (m, 3H), 1.94-1.57 (m, 3H), 1.48-1.20 (m, 5H). Additional signals of
minor rotamers were also
detected.
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Example 32
4-(1(2S,45)-24(9aR,11aR)-6-Chloro-3-oxo-3,9,9a,10,11,11a-hexahydro-2H-
cyc10but44,51 [3] benz -
oxocino [2,1-c] pyridin-2 -yll -4 -methoxypentanoyllamino)benzamide (single
stereoisomer)
0.0 H3
C H 3
+ 0
00J N H 2
= 0
Ci
1,1,3,3-Tetramethylguanidine (21 0.17 mmol, 3.0 eq.) was added under argon
atmosphere at RT
to a solution of
(9aR,11aR)-6-chloro-9a,10,11,11a-tetrahydro-2H-
cyclobuta[4,51[3]benzoxocino[2,1-clpyridin-3(91-1)-one (single stereoisomer)
(16 mg, 0.06 mmol) in
2-propanol / acetone (4:1, 0.6 m1). The mixture was stirred at RT for 15 min,
followed by addition
of 4-{[(2R,45)-2-bromo-4-methoxypentanoyllaminolbenzamide (single
stereoisomer) (18 mg, 0.06
mmol, 1.0 eq.) and 2-propanol / acetone (4:1, 0.6 m1). The reaction mixture
was stirred at RT for 3
days, followed by the addition of 1,1,3,3-tetramethylguanidine (7 jil, 0.06
mmol, 1.0 eq.), stirred at
RT for 1 additional day and concentrated under reduced pressure. The residue
was purified by
preparative HPLC (reversed phase, eluent: acetonitrile / water gradient).
Yield: 19 mg (61% of
theory).
LC-MS (method 4): R1= 1.86 min; MS (ESIpos): m/z = 536 [M+H1+
'H-NMR (600 MHz, DM50-d6): 6 [ppm] = 10.74-10.58 (m, 1H), 7.88-7.80 (m, 3H),
7.77-7.66 (m,
2H), 7.56-7.31 (m, 3H), 7.23 (br s, 1H), 6.40-6.35 (m, 1H), 5.84 (br t, 1H),
4.60 (q, 1H), 3.3-3.20 (m,
2H, partially concealed), 3.17 / 3.15 (2s, 3H), 2.79-2.66 (m, 2H), 2.64-2.56
(m, 2H), 2.32-2.04 (m,
3H), 1.82-1.66 (m, 1H), 1.40-1.22 (m, 1H), 1.19-1.10 (m, 3H). Additional
signals of minor rotamers
were also detected.
Example 33
4-(1(25)-24(7R)-11-Chloro-2-oxo-7-(trifluoromethyl)-2,6,7,8-tetrahydro-3H-
[3]benzoxocino [2,1-
c] pyridin-3 -yll butanoyl}amino)-2-fluorobenzamide (single stereoisomer)
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C H 3
FF
0
N \-11
0
N H 2
= 0
CI
1,1,3,3-Tetramethylguanidine (75 0.60 mmol, 3.0 eq.) was added under argon
atmosphere at RT
to a solution of (7 R) - 11-chloro-7-(trifluoromethyl)-7,8-dihydro-3H-
[3]benzoxocinop,1-clpyridin-
2(611)-one (single stereoisomer) (69 mg, 0.20 mmol) in 2-propanol / acetone
(4:1, 1.05 m1). The
mixture was stirred at RT for 15 min, followed by addition of 4-{[(2R)-2-
bromobutanoyllamino}-2-
fluorobenzamide (single stereoisomer) (79 mg, 0.26 mmol, 1.3 eq.) and of
further 2-propanol /
acetone (4:1, 1.05 m1). The reaction mixture was stirred at RT overnight and
concentrated under
reduced pressure. The residue was suspended in a mixture of NN-
dimethylformamide / acetonitrile
/ water. Material not dissolved was filtered and dried in vacuo . Yield: 52 mg
(47% of theory).
The filtrate was purified by preparative HPLC (reversed phase, eluent:
acetonitrile / water gradient).
Yield: 40 mg (36% of theory).
52 mg-batch: LC-MS (method 4): Rt = 1.93 min; MS (ESIpos): m/z = 552 [M+H1+
40 mg-batch: LC-MS (method 1): Rt = 1.04 min; MS (ESIpos): m/z = 552 [M+H1+
40 mg-batch: 'H-NMR (400 MHz, DM50-d6): 6 [ppm] = 10.93-10.77 (m, 1H), 8.00-
7.90 (m, 1H),
7.69 (t, 1H), 7.64 (dd, 1H), 7.59-7.47 (m, 3H), 7.47-7.32 (m, 3H), 6.47-6.35
(m, 1H), 5.56 (dd, 1H),
4.80-4.64 (m, 1H), 4.12-3.99 / 3.85-3.72 (2m, 1H), 3.27-3.19 / 3.15-2.99 (2m,
2H), 2.85-2.75 /2.61-
2.5 (2m, 1H, partially concealed), 2.25-2.08 (m, 2H), 0.99-0.84 (m, 3H).
Additional signals of minor
rotamers were also detected.
Example 34
4-( { (25)-2 -R7R)- 11-Chloro-2-oxo-7-(trifluoromethyl)-2,6,7,8-tetrahydro-3H-
[3]benzoxocino [2,1-
c] pyridin-3 -yll butanoyl}amino)benzamide (single stereoisomer)
C H 3
0 ENI
N
0 0
0
N H 2
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1,1,3,3-Tetramethylguanidine (38 0.30 mmol, 3.0 eq.) was added under argon
atmosphere at RT
to a solution of (7 R) - 11-chloro-7-(trifluoromethyl)-7,8-dihydro-3H-
[3]benzoxocinop,1-clpyridin-
2(611)-one (single stereoisomer) (34 mg, 0.10 mmol) in 2-propanol / acetone
(3:1, 0.8 m1). The
mixture was stirred at RT for 15 min, followed by addition of 4-{ [(2R)-2-
bromobutanoyllamino}benzamide (single stereoisomer) (31 mg, 0.11 mmol, 1.1
eq.) and of further
2-propanol / acetone (3:1, 0.8 m1). The reaction mixture was stirred at RT
overnight and concentrated
under reduced pressure. The residue was purified by preparative HPLC (reversed
phase, eluent:
acetonitrile / water gradient). Yield: 41 mg (76% of theory).
LC-MS (method 4): R1= 1.85 min; MS (ESIpos): m/z = 534 [M+H1+
'H-NMR (400 MHz, DM50-d6): 6 [ppm] = 10.74-10.62 (m, 1H), 8.00-7.92 (m, 1H),
7.91-7.81 (m,
3H), 7.68 (d, 2H), 7.56-7.32 (m, 3H), 7.30-7.19 (m, 1H), 6.47-6.34 (m, 1H),
5.60 (dd, 1H), 4.82-4.65
(m, 1H), 4.12-4.00 / 3.92-3.72 (2m, 1H), 3.3-3.19 / 3.16-2.96 (2m, 2H), 2.85-
2.75 / 2.62-2.5 (2m,
1H, partially concealed), 2.25-2.08 (m, 2H), 0.99-0.83 (m, 3H). Additional
signals of minor rotamers
were also detected.
Example 35
(25)-2 -[(7R)-11 -Chloro-2-oxo-7-(trifluoromethyl)-2,6,7, 8-tetrahydro-3H-
[3]benzoxocino [2,1-
clpyridin-3-y11-N-(2-methyl-2H-benzotriazol-5-yl)butanamide (single
stereoisomer)
C H3
0
N-- =
N¨C H 3
0
0
CI
1,1,3,3-Tetramethylguanidine (38 0.30 mmol, 3.0 eq.) was added under argon
atmosphere at RT
to a solution of (7 R) - 11-chloro-7-(trifluoromethyl)-7,8-dihydro-3H-
[3]benzoxocinop,1-clpyridin-
2(611)-one (single stereoisomer) (34 mg, 0.10 mmol) in 2-propanol / acetone
(3:1, 0.8 m1). The
mixture was stirred at RT for 15 min, followed by addition of (2R)-2-bromo-N-
(2-methy1-2H-
benzotriazol-5-yl)butanamide (single stereoisomer) (34 mg, 0.11 mmol, 1.1 eq.)
and of further 2-
propanol / acetone (3:1, 0.8 m1). The reaction mixture was stirred at RT
overnight and concentrated
under reduced pressure. The residue was purified by preparative HPLC (reversed
phase, eluent:
acetonitrile / water gradient). Yield: 33 mg (60% of theory).
LC-MS (method 4): Rt = 2.06 min; MS (ESIpos): m/z = 546 [M+HI
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'H-NMR (600 MHz, DMSO-d6): 6 [ppm] = 10.77-10.66 (m, 1H), 8.37-8.31 (m, 1H),
8.05-7.96 (m,
1H), 7.89 (d, 1H), 7.57-7.32 (m, 4H), 6.48-6.35 (m, 1H), 5.63 (dd, 1H), 4.81-
4.66 (m, 1H), 4.45 (s,
3H), 4.10-4.02 / 3.85-3.76 (2m, 1H), 3.14-2.96 (m, 2H), 2.64-2.52 (m, 1H,
partially concealed), 2.26-
2.08 (m, 2H), 0.99-0.84 (m, 3H). Additional signals of minor rotamers were
also detected.
Example 36
(25)-2 -[(7R)-11 -Chloro-2-oxo-7-(trifluoromethyl)-2,6,7, 8-tetrahydro-3H-
[3]benzoxocino [2,1-
clpyridin-3-y11-N-P-(difluoromethyl)-2H-indazol-5-yllbutanamide (single
stereoisomer)
C H 3
0
N
0
0
CI
General Method 7 was carried out with (7R)-11-chloro-7-(trifluoromethyl)-7,8-
dihydro-3H-
.. [3]benzoxocino[2,1-c]pyridin-2(6H)-one (single stereoisomer) (33.9 mg, 103
umol, 1.0 eq.), (2R)-
2-bromo-N42-(difluoromethyl)-2H-indazol-5-yllbutanamide (single stereoisomer)
(57.0 mg, 90%
purity, 154 umol, 1.5 eq.) and 1,1,3,3-tetramethylguanidine (37 IA, 290 umol,
2.84 eq.) in a mixture
of 2-propanol / acetone (4:1, 1.25 ml) including the following variations of
the procedure: After
stirring of the reaction mixture at RT overnight, further amounts of 1,1,3,3-
tetramethylguanidine (19
150 umol, 1.5 eq.) were added and the mixture was stirred again overnight. The
crude mixture
was purified by preparative HPLC (reversed phase, eluent: water with 0.05%
formic acid /
acetonitrile 90:10 to 1:95). Yield: 23.0 mg (38% of theory).
LC-MS (method 3): R1= 3.80 min; MS (ESIpos): m/z = 581 [M+H1+
1H-NMR (500 MHz, DM50-d6): 6 [ppm] = 10.61 / 10.57 (2br s, 1H), 8.80 (s, 1H),
8.33-8.23 (m,
1H), 8.22 / 8.10 (2s, 1H), 8.04-7.94 (m, 1H), 7.72 (d, 1H), 7.57-7.47 (m, 1H),
7.49-7.35 (m, 3H),
6.44 / 6.39 (2s, 1H), 5.72-5.52 (m, 1H), 4.85-4.64 (m, 1H), 3.88 / 3.79 (2t,
1H), 3.27-2.93 (m, 2H),
2.63-2.53 (m, 1H), 2.24-2.15 (m, 1H), 2.15-2.03 (m, 1H), 0.94 / 0.91 (2t, 3H).
Additional signals of
minor rotamers were also detected.
Example 37
(25)-2 -[(7R)-11 -Chloro-2-oxo-7-(trifluoromethyl)-2,6,7, 8-tetrahydro-3H-
[3]benzoxocino [2,1-
clpyridin-3-y11-N-P-(trifluoromethyl)-2H-indazol-5-yllbutanamide (single
stereoisomer)
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C H 3
0 1-1\11
N
F
0
0 N F
=
C I
General Method 7 was carried out with (7R)-11-chloro-7-(trifluoromethyl)-7,8-
dihydro-3H-
[3]benzoxocino[2,1-clpyridin-2(6H)-one (single stereoisomer) (71.7 mg, 217
umol, 1.0 eq.), (2R)-
2-bromo-N42-(trifluoromethyl)-2H-indazol-5-yllbutanamide (single stereoisomer)
(177 mg, 86%
purity, 435 umol, 2.0 eq.) and 1,1,3,3-tetramethylguanidine (77 IA, 620 umol,
2.83 eq.) in a mixture
of 2-propanol / acetone (4:1, 3.8 ml) including the following variations of
the procedure: After
stirring of the reaction mixture at RT overnight, further amounts of 1,1,3,3-
tetramethylguanidine (41
IA, 330 umol, 1.5 eq.) were added and the mixture was stirred again overnight.
The crude mixture
was purified by preparative HPLC (reversed phase, eluent: water with 0.05%
formic acid /
acetonitrile 90:10 to 1:95). Yield: 86.0 mg (66% of theory).
LC-MS (method 3): R1= 4.13 min; MS (ESIpos): m/z = 599 [M+H1+
1H-NMR (500 MHz, DM50-d6): 6 [ppm] = 10.67 / 10.63 (2s, 1H), 9.07 / 9.06 (2s,
1H), 8.30 / 8.27
(2s, 1H), 7.99 / 7.96 (2s, 1H), 7.75 (d, 1H), 7.57-7.30 (m, 4H), 6.44 / 6.38
(2s, 1H), 5.67-5.57 (m,
1H), 4.81-4.64 (m, 1H), 3.79 (t, 1H), 3.17-2.92 (m, 2H), 2.60-2.52 (m, 1H),
2.26-2.14 (m, 1H), 2.14-
.. 2.03 (m, 1H), 0.94 / 0.91 (2t, 3H). Additional signals of minor rotamers
were also detected.
Example 38
4-( {2- [(7R)-11-Chloro-2-oxo-7-(trifluoromethyl)-2,6,7,8-tetrahydro-3H-
[3]benzoxocino [2,1-
c] pyridin-3 -yll pentanoyl}amino)benzamide (mixture of two diastereomers)
C H 3
0 1-N1
N
0 0
0
N H 2
CI
1,1,3,3-Tetramethylguanidine (35 IA, 0.28 mmol, 3.0 eq.) was added under argon
atmosphere at RT
to a solution of (7 R) - 11-chloro-7-(trifluoromethyl)-7,8-dihydro-3H-
[3]benzoxocinop,1-clpyridin-
2(611)-one (single stereoisomer) (31 mg, 0.09 mmol) in 2-propanol / acetone
(3:1, 0.8 m1). The
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mixture was stirred at RT for 15 min, followed by addition of 44(2-
bromopentanoyl)aminolbenzamide (racemate) (33 mg, 92% purity, 0.10 mmol, 1.1
eq.) and of
further 2-propanol / acetone (3:1, 0.8 m1). The reaction mixture was stirred
at RT overnight and
concentrated under reduced pressure. The residue was purified by preparative
HPLC (reversed phase,
eluent: acetonitrile / water gradient). Yield: 37 mg (73% of theory).
LC-MS (method 4): R1= 1.92 min; MS (ESIpos): m/z = 548 [M+H1+
'H-NMR (400 MHz, DM50-c/6): 6 [ppm] = 10.76-10.65 (m, 1H), 7.99 (d, 1H), 7.90-
7.80 (m, 3H),
7.68 (d, 2H), 7.57-7.32 (m, 3H), 7.30-7.20 (m, 1H), 6.46-6.35 (m, 1H), 5.75-
5.63 (m, 1H), 4.81-4.66
(m, 1H), 4.10-3.73 (m, 1H), 3.28-2.98 (m, 2H), 2.62-2.5 (m, 1H, partially
concealed), 2.21-2.02 (m,
2H), 1.43-1.19 (m, 2H), 0.94 (t, 3H). Additional signals of minor rotamers
were also detected.
Example 39
4- {2 -[(7R)-11-Chloro -2 -oxo-7-(trifluoromethyl)-2,6,7,8-tetrahydro-3H- [3]
benzoxocino [2,1-
c] pyridin-3 -yll -3 -cyclobutylpropanamido} -2-fluorobenzamide (mixture of
two diastereomers)
0
N
00 N H 2
0
CI
General Method 7 was carried out with (7R)-11-chloro-7-(trifluoromethyl)-7,8-
dihydro-3H-
[31benz-oxocino[2,1-clpyridin-2(6H)-one (single stereoisomer) (29.0 mg, 78%
purity, 68.5 umol,
1.0 eq.), 4-(2-bromo-3-cyclobutylpropanamido)-2-fluorobenzamide (racemate)
(31.0 mg, 91%
purity, 82.2 umol, 1.2 eq.), and 1,1,3,3-tetramethylguanidine (26
210 umol, 3.0 eq.) in a mixture
of 2-propanol / acetone (4:1, 1.25 ml) including the following variations of
the procedure: The crude
mixture was purified by preparative HPLC (reversed phase, eluent: water with
0.05% formic acid /
acetonitrile 90:10 to 5:95). Yield: 20.0 mg (49% of theory).
LC-MS (method 3): Rt = 3.89 / 3.94 min; MS (ESIpos): m/z = 592 [M+H1+
Example 40
4-( (25)-2 -[ (7 R) - 11-Chloro-2-oxo-7-(trifluoromethyl)-2,6,7,8-
tetrahydro-3H-[3]benzoxocino [2,1-
clpyridin-3-yll -3 -cyclobutylpropanoyl}amino)-2-fluorobenzamide (single
stereoisomer)
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F
0 c(l?1
N
00 10 N H 2
= 0
CI
Diastereomer separation of 15 mg of 4-{2-[(7R)-11-chloro-2-oxo-7-
(trifluoromethyl)-2,6,7,8-
tetrahydro-3H-[3]benzoxocino [2,1-c] pyridin-3 -yl] -3 -cyclobutylpropanamido}
-2 -fluorobenzamide
(mixture of two diastereomers), Example 39 gave
single stereoisomer 1 (the title compound 40) (chiral HPLC: R1= 4.66 min, 99%
de): 6.5mg (14% of
theory),
single stereoisomer 2 (chiral HPLC: R1 = 7.07 min): 8.8 mg.
Separation method: HPLC: column: Daicel Chiralpak ID 5 pm, 250 mm x 20 mm;
eluent: 55%
n-heptane / 45% 2-propanol; temperature: 40 C; flow rate: 20 ml/min; UV
detection: 220 nm.
Analysis method: HPLC: column: Daicel Chiralpak ID 5 pm, 250 mm x 4.6 mm;
eluent: 55%
iso-hexane / 45% 2-propanol; flow rate: 1.0 ml/min; UV detection: 220 nm.
LC-MS (method 5): R1= 1.45 min; MS (ESIneg): m/z = 590 EM-F11-
1H-NMR (400 MHz, DM50-d6): 6 [ppm] = 10.87 / 10.80 (2s, 1H), 7.96 (s, 1H),
7.72-7.67 (m, 1H),
7.67-7.62 (m, 1H), 7.57-7.35 (m, 6H), 6.41 / 6.37 (2s, 1H), 5.66-5.54 (m, 1H),
4.81-4.62 (m, 1H),
3.78 (t, 1H), 3.07 (br d, 2H), 2.60-2.55 (m, 1H), 2.27-2.15 (m, 3H), 2.02-1.88
(m, 2H), 1.84-1.73 (m,
3H), 1.67-1.58 (m, 1H). Additional signals of minor rotamers were also
detected.
Example 41
4- {2 -R7R)-11-Chloro -2 -oxo-7-(trifluoromethyl)-2,6,7,8-tetrahydro-3H- [3]
benzoxocino [2,1-
c] pyridin-3 -yll -3 4(25)-S ,5 -dimethyltetrahydrofuran-2 -yll
propanamidolbenzamide (mixture of two
diastereomers)
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H 3C
O2çCH3
0
N
0 N H 2
0
Ci
2-[(7R)-11 -Chloro -2 -oxo -7 -(trifluoromethyl)-2,6,7,8-tetrahydro-3H-
[3]benzoxocino [2,1-c] pyridin-
3-yll -3-[(2S)-5,5-dimethyloxolan-2-yllpropanoic acid (mixture of two
diastereomers) (98 mg, 87%
purity, 0.17 mmol), pyridine (15 0.19 mmol, 1.1 eq.) and T3P (152 IA, 50%
solution in ethyl
acetate, 0.26 mmol, 1.5 eq.) were added under argon atmosphere at RT to a
solution of 4-
aminobenzamide (26 mg, 0.19 mmol, 1.1 eq.) in tetrahydrofuran (5 m1). The
reaction mixture was
stirred at RT for 1.5 h and put into iced water. After phase separation, the
organic phase was extracted
with ethyl acetate. The combined organic phases were washed with brine, dried
over anhydrous
sodium sulfate, filtered and concentrated under reduced pressure. The residue
was purified by
preparative HPLC (reversed phase, eluent: acetonitrile / water gradient).
Yield: 67 mg (64% of
theory).
LC-MS (method 1): R1= 1.08 min; MS (ESIpos): m/z = 618 [M+H1+
'H-NMR (600 MHz, DM50-c/6): 6 [ppm] = 10.72-10.52 (m, 1H), 8.02-7.95 (m, 1H),
7.87 (br s, 1H),
7.84 (d, 2H), 7.69(d, 2H), 7.56-7.31 (m, 3H), 7.25 (br s, 1H), 6.44-6.32 (m,
1H), 5.85-5.60 (m, 1H),
4.80-4.64 (m, 1H), 4.09-3.70 (m, 2H), 3.14-2.99 (m, 2H), 2.79-2.70 / 2.60-2.5
(2m, 1H, partially
concealed), 2.42-2.19 (m, 2H), 2.04-1.94 (m, 1H), 1.74-1.55 (m, 3H), 1.15 /
1.09 (2s, 6H). Additional
signals of minor rotamers were also detected.
Example 42
4-( { (25)-2 -R7R)- 11-Chloro-2-oxo-7-(trifluoromethyl)-2,6,7,8-tetrahydro-3H-
[3]benzoxocino [2,1-
clpyridin-3-yll -34(25)-5,5 -dimethyloxolan-2-yllpropanoyl}amino)benzamide
(single stere oi some r)
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H 3C
es.r\.1.1 C H 3
0
0
N'(
00 N H 2
CI
Diastereomer separation of 64 mg of 4-{2-[(7R)-11-chloro-2-oxo-7-
(trifluoromethyl)-2,6,7,8-
tetrahydro-3H-[3]benzoxocino [2, 1-c] pyridin-3 -yl] -3 -{(25)-5 ,5 -
dimethyltetrahydrofuran-2 -
yllpropanamido}-benzamide (mixture of two diastereomers), Example 41 gave
single stereoisomer 1 (the title compound Example 42) (chiral HPLC: Rt = 2.50
min, >99% de): 47
mg,
single stereoisomer 2 (chiral HPLC: Rt = 3.90 min, >99% de): 10 mg.
Separation method: HPLC: column: Daicel Chiralpak OZ-H 5 pm, 250 mm x 20 mm;
eluent: 70%
n-heptane / 30% ethanol; temperature: 40 C; flow rate: 25 ml/min; UV
detection: 220 nm.
Analysis method: HPLC: column: Daicel Chiralpak OZ-3 3 pm, 50 mm x 4.6 mm;
eluent: 80%
n-hexane / 20% ethanol; temperature: 40 C; flow rate: 1 ml/min; UV detection:
220 nm.
'H-NMR (500 MHz, DM50-d6): 6 [ppm] = 10.72-10.50 (m, 1H), 7.97 (s, 1H), 7.91-
7.80 (m, 3H),
7.68 (d, 2H), 7.57-7.32 (m, 3H), 7.24 (br s, 1H), 6.45-6.32 (m, 1H), 5.78-5.60
(m, 1H), 4.81-4.64 (m,
1H), 4.09-3.71 (m, 2H), 3.14-2.99 (m, 2H), 2.81-2.69 / 2.60-2.5 (2m, 1H,
partially concealed), 2.42-
2.18 (m, 2H), 2.04-1.93 (m, 1H), 1.75-1.55 (m, 3H), 1.15 / 1.09 (2s, 6H).
Additional signals of minor
rotamers were also detected.
Example 43
4-( (25)-2 -[(7R)-11 -Chloro-2-oxo -7-(trifluoromethyl)-2,6,7,8-tetrahydro-3H-
[3]benzoxocino [2,1-
c] pyridin-3 -yll -3 - [(25)-tetrahydro-2H-pyran-2-yll propanoyllamino)-2-
fluorobenzamide (single
stereoisomer)
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s .110
0
N
0 el
0
I N H 2
CI
1,1,3,3-Tetramethylguanidine (45
0.36 mmol, 3.0 eq.) was added under argon atmosphere at RT
to a solution of (7 R) - 11-chloro-7-(trifluoromethyl)-7,8-dihydro-3H-
[3]benzoxocinop,1-clpyridin-
2(611)-one (single stereoisomer) (40 mg, 0.12 mmol) in 2-propanol / acetone
(3:1, 0.8 m1). The
mixture was stirred at RT for 15 min, followed by addition of 4-({(2R)-2-bromo-
3-[(25)-tetrahydro-
2H-pyran-2-yllpropanoyl}amino)-2-fluorobenzamide (single stereoisomer) (49 mg,
0.13 mmol, 1.1
eq.) and of further 2-propanol / acetone (3:1, 0.8 m1). The reaction mixture
was stirred at RT
overnight. Further 1,1,3,3-Tetramethylguanidine (15
0.12 mmol, 1.0 eq.) was added. The reaction
mixture was stirred for 4 h and concentrated under reduced pressure. The
residue was purified by
preparative HPLC (reversed phase, eluent: acetonitrile / water gradient).
Yield: 41 mg (55% of
theory).
LC-MS (method 4): Rt = 2.08 min; MS (ESIpos): m/z = 622 [M+H1+
'H-NMR (400 MHz, DM50-d6): 6 [ppm] = 10.89-10.62 (m, 1H), 8.03-7.87 (m, 1H),
7.72-7.60 (m,
2H), 7.59-7.32 (m, 6H), 6.44-6.32 (m, 1H), 5.82-5.61 (m, 1H), 4.80-4.64 (m,
1H), 4.13-4.00 / 3.91-
3.72 (2m, 2H), 3.3-3.18 (m, 2H, partially concealed), 3.18-2.97 (m, 2H), 2.62-
2.5 (m, 1H, partially
concealed), 2.31-2.08 (m, 2H), 1.83-1.70 (m, 1H), 1.67-1.56 (m, 1H), 1.49-1.19
(m, 4H). Additional
signals of minor rotamers were also detected.
Example 44
4-( { (25)-2 -R7R)- 11-Chloro-2-oxo-7-(trifluoromethyl)-2,6,7,8-tetrahydro-3H-
[3]benzoxocino [2,1-
clpyridin-3-yll -3 - [(25)-tetrahydro-2H-pyran-2-yll propanoyllamino)benzamide
(single
stereoisomer)
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0
N
0J NH 2
0
"CI
1,1,3,3-Tetramethylguanidine (75 0.60 mmol, 3.0 eq.) was added under argon
atmosphere at RT
to a solution of (7 R) - 11-chloro-7-(trifluoromethyl)-7,8-dihydro-3H-
[3]benzoxocinop,1-clpyridin-
2(611)-one (single stereoisomer) (69 mg, 0.20 mmol) in 2-propanol / acetone
(4:1, 2.5 m1). The
mixture was stirred at RT for 15 min, followed by addition of 4-({(2R)-2-bromo-
3-[(25)-tetrahydro-
2H-pyran-2-yllpropanoyl}amino)benzamide (single stereoisomer) (78 mg, 0.22
mmol, 1.1 eq.) and
of further 2-propanol / acetone (4:1, 2.5 m1). The reaction mixture was
stirred at RT for 4 days and
concentrated under reduced pressure. The residue was purified by preparative
HPLC (reversed phase,
eluent: acetonitrile / water gradient). Yield: 77 mg (64% of theory).
LC-MS (method 4): R1= 1.97 min; MS (ESIpos): m/z = 604 [M+H1+
'H-NMR (500 MHz, DM50-d6): 6 [ppm] = 10.72-10.52 (m, 1H), 8.05-7.80 (m, 4H),
7.73-7.66 (m,
2H), 7.56-7.32 (m, 3H), 7.31-7.22 (m, 1H), 6.45-6.32 (m, 1H), 5.85-5.66 (m,
1H), 4.79-4.66 (m, 1H),
4.10-4.01 / 3.90-3.73 (2m, 2H), 3.3-3.20 (m, 2H), 3.17-2.98 (m, 2H), 2.62-2.5
(m, 1H, partially
concealed), 2.32-2.10 (m, 2H), 1.81-1.71 (m, 1H), 1.67-1.57 (m, 1H), 1.48-1.20
(m, 4H). Additional
signals of minor rotamers were also detected.
Example 45
4-( { (45)-2 -[(7R)-11 -Chloro-2-oxo -7-(trifluoromethyl)-2,6,7,8-tetrahydro-
3H-[3]benzoxocino [2,1-
c] pyridin-3 -yll -4-methoxypentanoyl}amino)-2-fluorobenzamide (mixture of two
diastereomers)
C H3
C H 3
0
N
00 el N H 2
CI
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- 201 -4-Amino-2-fluorobenzamide (38 mg, 0.25 mmol, 1.1 eq.), /V,N-
diisopropylethylamine (97 IA,
0.56 mmol, 2.5 eq.) and a solution of HATU (110 mg, 0.29 mmol, 1.3 eq.) in
/V,N-
dimethylformamide (2 ml) were added under argon atmosphere at RT to a solution
of (2)-2-R7R)-
11 -chloro-2-oxo-7-(trifluoromethyl) -2,6,7, 8-tetrahydro-3H-[3] benzoxocino
[2,1 -c] pyridin-3 -yl] -
2,3,5-trideoxy-4-0-methyl-L-glycero-pentonic acid (mixture of two
diastereomers) (128 mg, 80%
assumed purity of crude product, 0.22 mmol) in NN-dimethylformamide (4 m1).
The reaction
mixture was stirred at RT for 5.5 h and concentrated under reduced pressure.
The residue was
crystallized with water, filtered, washed with water and dried in vacuo. This
residue was dissolved
in dichloromethane and purified by column chromatography (silica gel, eluent:
dichloromethane /
methanol gradient). Yield: 52 mg (39% of theory).
LC-MS (method 4): R1= 1.95 min; MS (ESIpos): m/z = 596 [M+H1+
'H-NMR (600 MHz, DM50-d6): 6 [ppm] = 10.92-10.73 (m, 1H), 8.06-7.97 (m, 1H),
7.72-7.61 (m,
2H), 7.59-7.33 (m, 6H), 6.46-6.34 (m, 1H), 5.84-5.71 (m, 1H), 4.79-4.66 (m,
1H), 4.08-3.95 / 3.92-
3.74 (2m, 1H), 3.3-3.00 (m, 6H, partially concealed), 2.83-2.74 / 2.62-2.5
(2m, 1H, partially
concealed), 2.40-2.16 (m, 2H), 1.19-1.08 (m, 3H). Additional signals of minor
rotamers were also
detected.
Example 46
4-( { (2S,45)-2-[(7R)-11-Chl oro-2-oxo-7-(trifluoromethyl)-2,6,7,8 -tetrahydro-
3H-
[3] benzoxocino [2,1 -c] pyridin-3 -yl] -4-methoxypentanoyllamino) -2 -
fluorobenzamide (single
stereoisomer)
CH3
0
N
00 NH 2
0
CI
Diastereomer separation of 50 mg of 4-({(45)-2-R7R)- 11-chloro-2-oxo-7-
(trifluoromethyl)-2,6,7,8-
tetrahydro-3H-[3]benzoxocino 112, 1 -c] pyridin-3 -yl] -4 -
methoxypentanoyl}amino)-2 -
fluorobenzamide (mixture of two diastereomers), Example 45 gave
single stereoisomer 1 (the title compound Example 46) (chiral SFC: Rt = 0.61
min, >99% de): 17
mg,
single stereoisomer 2 (chiral SFC: Rt = 0.86 min, >99% de): 21 mg.
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Separation method: SFC: column: Daicel Chiralpak IG 5 um, 250 mm x 20 mm;
eluent: 70% carbon
dioxide / 30% methanol; temperature: 40 C; flow rate: 80 ml/min; UV detection:
210 nm.
Analysis method: SFC: column: Daicel Chiralpak IG 3 um, 50 mm x 4.6 mm;
eluent: 70% carbon
dioxide / 30% 2-propanol; temperature: 40 C; flow rate: 3 ml/min; UV
detection: 220 nm.
'H-NMR (400 MHz, DMSO-d6): 6 [ppm] = 10.92-10.71 (m, 1H), 8.05-7.96 (m, 1H),
7.73-7.60 (m,
2H), 7.58-7.32 (m, 6H), 6.46-6.33 (m, 1H), 5.84-5.68 (m, 1H), 4.80-4.64 (m,
1H), 4.10-3.97 / 3.84-
3.72 (2m, 1H), 3.3-2.98 (m, 6H, partially concealed), 2.84-2.73 / 2.63-2.5
(2m, 1H, partially
concealed), 2.31-2.16 (m, 2H), 1.15 (d, 3H). Additional signals of minor
rotamers were also detected.
Example 47
4-( { (45)-2 -R7R)- 11-Chloro-2-oxo-7-(trifluoromethyl)-2,6,7,8-tetrahydro-3H-
[3]benzoxocino [2,1-
c] pyridin-3 -yll -4-methoxypentanoyl}amino)benzamide (mixture of two
diastereomers)
C H3
C H3
0
N
00 N H 2
=
CI
4-Aminobenzamide (33 mg, 0.25 mmol, 1.1 eq.), NN-diisopropylethylamine (97 IA,
0.56 mmol, 2.5
eq.) and a solution of HATU (110 mg, 0.29 mmol, 1.3 eq.) in NN-
dimethylformamide (2 ml) were
added under argon atmosphere at RT to a solution of (2)-2-[(7R)-11-chloro-2-
oxo-7-
(trifluoromethyl)-2,6,7,8-tetrahydro-3H-[3]benzoxocino [2,1-c] pyridin-3 -yll -
2,3,5 -trideoxy-4 -0-
methyl-L-glycero-pentonic acid (mixture of two diastereomers) (128 mg, 80%
assumed purity of
crude product, 0.22 mmol) in NN-dimethylformamide (4 m1). The reaction mixture
was stirred at
RT for 2 h and concentrated under reduced pressure. The residue was
crystallized with water, filtered,
washed with water and dried in vacuo. This residue was purified by column
chromatography (silica
gel, eluent: cyclohexane / ethyl acetate gradient). Yield: two batches
isolated, 67 mg and 16 mg (64%
of theory for both batches together).
67 mg-batch: LC-MS (method 4): Rt = 1.88 min; MS (ESIpos): m/z = 578 [M+F11+
Example 48
4-( {(2S,45)-2-R7R)- 11-Chl oro-2-oxo-7-(trifluoromethyl)-2,6,7,8 -tetrahydro-
3H-
[3] benzoxocino [2,1-c] pyridin-3 -yl] -4-methoxypentanoyllamino)benzamide
(single stereoi some r)
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- 203 -0'C H3
C H3
0
N
00 N H 2
0
Ci
Diastereomer separation of 82 mg of 4-({(45)-24(7R)- 11-chloro-2-oxo-7-
(trifluoromethyl)-2,6,7,8-
tetrahydro-3H-[3]benzoxocino [2, 1 -c] pyridin-3 -yl] -4 -
methoxypentanoyl}amino)benzamide
(mixture of two diastereomers), Example 47 gave
single stereoisomer 1 (the title compound Example 48) (chiral SFC: Rt = 4.44
min, >99% de): 19
mg,
single stereoisomer 2 (chiral SFC: Rt = 6.31 min, >99% de): 22 mg.
Separation method: SFC: column: Daicel Chiralpak IE 5 um, 250 mm x 20 mm;
eluent: 65% carbon
dioxide / 35% methanol; temperature: 40 C; flow rate: 80 ml/min; UV detection:
210 nm.
Analysis method: SFC: column: Daicel Chiralpak IE, 50 mm x 4.6 mm; eluent: 70%
carbon dioxide /
30% methanol; temperature: 40 C; flow rate: 3 ml/min; UV detection: 210 nm.
'H-NMR (400 MHz, DM50-d6): 6 [ppm] = 10.75-10.58 (m, 1H), 8.05-7.97 (m, 1H),
7.91-7.79 (m,
3H), 7.70 (d, 2H), 7.57-7.32 (m, 3H), 7.25 (br s, 1H), 6.46-6.33 (m, 1H), 5.86-
5.73 (m, 1H), 4.80-
4.64 (m, 1H), 4.10-3.95 /3.84-3.71 (2m, 1H), 3.3-2.97 (m, 6H, partially
concealed), 2.85-2.73 /2.63-
2.5 (2m, 1H, partially concealed), 2.30-2.15 (m, 2H), 1.16 (d, 3H). Additional
signals of minor
rotamers were also detected.
Alternative synthetic route:
1,1,3,3-Tetramethylguanidine (93 0.74 mmol, 3.0 eq.) was added under argon
atmosphere at RT
to a solution of (7 R) - 11-chloro-7-(trifluoromethyl)-7,8-dihydro-3H-
[3]benzoxocinop,1-clpyridin-
2(611)-one (single stereoisomer) (82 mg, 0.25 mmol) in 2-propanol / acetone
(4:1, 2 m1). The mixture
was stirred at RT for 15 min, followed by addition of 4-{[(2R,45)-2-bromo-4-
methoxypentanoyllamino}benzamide (single stereoisomer) (90 mg, 0.27 mmol, 1.1
eq.) and of
further 2-propanol / acetone (4:1, 2 m1). The reaction mixture was stirred at
RT for 5 days and
concentrated under reduced pressure. The residue was purified by preparative
HPLC (reversed phase,
eluent: acetonitrile / water gradient). Yield: 106 mg (74% of theory).
LC-MS (method 1): R1= 1.00 min; MS (ESIpos): m/z = 578 [M+F11+
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'FI-NMR (400 MHz, DMSO-d6): 6 [ppm] = 10.75-10.56 (m, 1H), 8.07-7.97 (m, 1H),
7.92-7.80 (m,
3H), 7.70 (d, 2H), 7.57-7.32 (m, 3H), 7.25 (br s, 1H), 6.46-6.33 (m, 1H), 5.88-
5.73 (m, 1H), 4.81-
4.64 (m, 1H), 4.10-3.97 / 3.92-3.72 (2m, 1H), 3.3-2.97 (m, 6H, partially
concealed), 2.87-2.73 /2.63-
2.5 (2m, 1H, partially concealed), 2.30-2.15 (m, 2H), 1.16 (d, 3H). Additional
signals of minor
rotamers were also detected.
Further alternative synthetic route:
1,1,3,3-Tetramethylguanidine (2.19 ml, 17.5 mmol, 3.0 eq.) was added under
argon atmosphere at
RT to a solution of (7 R) - 11-chloro-7-(trifluoromethyl)-7,8-dihydro-3H-
[3]benzoxocino [2,1-
clpyridin-2(6H)-one (single stereoisomer) (2.00 g, 96% purity, 5.82 mmol) in 2-
propanol / acetone
(4:1, 136 m1). The mixture was stirred at RT for 15 min, followed by addition
of 4-{[(2R,45)-2-
bromo-4-methoxypentanoyllamino}benzamide (single stereoisomer) (3.26 g, 9.90
mmol, 1.7 eq.).
The reaction mixture was stirred at RT for 3 days and then poured into 0.05 N
aqueous hydrogen
chloride solution (700 m1). After extraction with ethyl acetate (2 x 100 ml),
the combined organic
phases were washed with saturated aqueous sodium chloride solution (100 ml),
dried over anhydrous
sodium sulfate, filtered and concentrated under reduced pressure. The residue
was purified by column
chromatography (Biotage Isolera, 100 g SNAP-Ultra cartridge, eluent:
dichloromethane (5 cv),
dichloromethane / acetonitrile 75:25, 50:50, 25:75 (5 cv each), acetonitrile
(20 cv) to yield two
fractions. Yield: 2.00 g (59% of theory, >98% de), 1.00 g (29% of theory, >49%
de).
Chiral SFC: Rt = 4.03 min, >98% de
Analysis method: SFC: column: Chiralcel AZ-3, 3 um, 100 mm x 4.6 mm, eluent:
isocratic 80%
carbon dioxide / 20% isopropanol, 10 min, temperature: 40 C; flow rate: 3.0
ml/min; UV detection:
210 nm, backpressure: 130 bar.
Further alternative synthetic route:
1,1,3,3-Tetramethylguanidine (68.5 ml, 546 mmol, 3.0 eq.) was added under
argon atmosphere at
RT to a solution of (7 R) - 11-chloro-7-(trifluoromethyl)-7,8-dihydro-3H-
[3]benzoxocino [2,1-
c]pyridin-2(6H)-one (single stereoisomer) (60.0 g, 182 mmol) in 2-propanol /
acetone (4:1, 2.1 1).
The mixture was stirred at RT for 15 min, followed by addition of 4-{[(2R,45)-
2-bromo-4-
methoxypentanoyllamino}benzamide (single stereoisomer) (89.9 g, 273 mmol, 1.5
eq.). The reaction
mixture was stirred at RT for 3 days and then neutralized with acidic acid
(31.3 ml, 546 mmol,
3.0 eq.). The reaction was concentrated to 1/4 of the original volume and then
poured into 0.02 N
aqueous hydrogen chloride solution (4.5 1). The precipitate was filtered off
under reduced pressure,
washed with water and dried under air atmosphere (yield: 119 g). The crude
material was purified
via SFC to obtain the product as single stereoisomer. Yield: 77.8 g (73% of
theory, >99% de).
Separation method: SFC: column: Chiralpak AZ 20 um, 350 mm x 50 mm, eluent:
isocratic 70%
carbon dioxide / 30% isopropanol, 4 min, temperature: 30 C; flow rate: 300
ml/min; UV detection:
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210 nm; backpressure: 100 bar.
Chiral SFC: R1= 1.39 min, >99% de
Analysis method: SFC: column: Chiralcel AZ-3, 3 p.m, 100 mm x 4.6 mm, eluent:
isocratic 70%
carbon dioxide / 30% isopropanol, 10 min, temperature: 40 C; flow rate: 3.0
ml/min; UV detection:
210 nm, backpressure: 130 bar.
Chiral HPLC: Rt = 3.15 min, >99% de
Analysis method: HPLC: column: Chiralcel OZ-3, 3 p.m, 50 mm x 4,6 mm, eluent:
isocratic 80%
heptane /20% ethanol, 10 min, temperature: 50 C; flow rate: 1.0 ml/min; UV
detection: 210 nm.
LC-MS (method 3): R1= 3.12 min; MS (ESIpos): m/z = 578 [M+H1+
1H-NMR (600 MHz, DM50-c/6): 6 [ppm] = 10.70-10.53 (m, 1H), 8.05-7.94 (m, 1H),
7.85 (br d, 3H),
7.70 (d, 2H), 7.58-7.32 (m, 3H), 7.21 (br s, 1H), 6.48-6.31 (m, 1H), 5.91-5.71
(m, 1H), 4.83-4.64 (m,
1H), 4.09-3.97 / 3.87-3.68 (2m, 1H), 3.39-2.99 (m, 6H, partially concealed),
2.85-2.74 / 2.65-2.50
(2m, 1H, partially concealed), 2.34-2.15 (m, 2H), 1.16 (d, 3H). Additional
signals of minor rotamers
were also detected.
13C-NMR (126 MHz, DM50-c/6): 6 [ppm] = 168.03 (s, 1C), 167.17 (s, 1C), 159.71
(s, 1C), 146.90
(s, 1C), 140.97 (2 overlapped s, 2C), 136.48 (s, 1C), 135.92 (s, 1C), 131.89
(s, 1C), 131.62 (d, 1CH),
129.74 (d, 1CH), 129.33 (overlapped d, 1CH), 129.33 (overlapped s, 1C), 128.27
(d, 2CH), 128.02
(d, 1CH), 126.24 [s, 1CF3 (J=280.8 Hz)], 118.78 (d, 2CH), 116.67 (d, 1CH),
75.08 (t, 1CH2), 73.54
(d, 1CH), 55.45 (q, 1CH30), 55.37 (d, 1CH), 44.23 (d, 1CH), 37.33 (t, 1CH2),
29.72 (t, 1CH2), 18.55
(q, 1CH3).
Example 49
(45)-2 -[(7R)-11 -Chloro-2-oxo-7-(trifluoromethyl)-2,6,7,8-tetrahydro-3H-
[3]benzoxocino [2,1-
clpyridin-3-y11-4-methoxy-N-([1,2,41triazolo[1,5-alpyridin-7-yl)pentanamide
(mixture of two
diastereomers)
WC H3
C H3
0
0 N,Na
0
CI
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[1,2,41Triazolo[1,5-alpyridin-7-amine hydrochloride (23 mg, 0.13 mmol, 1.1
eq.), pyridine (28
0.35 mmol, 3.0 eq.) and T3P (102 jil, 50% solution in ethyl acetate, 0.17
mmol, 1.5 eq.) were added
under argon atmosphere at RT to a solution of (2)-2-[(7R)-11-chloro-2-oxo-7-
(trifluoromethyl)-
2,6,7,8-tetrahydro-3H-[3]benzoxocino [2,1 -c] pyridin-3 -yl] -2,3 ,5 -tride
oxy-4-0-methyl-L -glyce ro -
__ pentonic acid (mixture of two diastereomers) (57 mg, 93% purity, 0.12 mmol)
in tetrahydrofuran
(4 m1). The reaction mixture was stirred at RT overnight before all volatiles
were removed under
reduced pressure. The residue was purified by preparative HPLC (reversed
phase, eluent: acetonitrile
/ water gradient). Yield: 44 mg (66% of theory).
LC-MS (method 1): Rt = 0.99 min; MS (ESIpos): m/z = 576 [M+I-11+
__ 'H-NMR (600 MHz, DM50-c/6): 6 [ppm] = 11.06-10.87 (m, 1H), 8.87 (d, 1H),
8.39 / 8.38 (2s, 1H),
8.22-8.16 (m, 1H), 8.09-7.91 (m, 1H), 7.56-7.29 (m, 4H), 6.46-6.36 (m, 1H),
5.87-5.74 (m, 1H),
4.80-4.69 (m, 1H), 4.11-3.75 (m, 1H), 3.3-2.98 (m, 3H, partially concealed),
3.19 / 3.12 (2s, 3H),
2.84-2.72 / 2.60-2.46 (2m, 1H, partially concealed), 2.43-2.20 (m, 2H), 1.20-
1.11 (m, 3H). Additional
signals of minor rotamers were also detected.
Example 50
(2S,4S)-2 -[(7R)-11 -Chloro -2-oxo -7-(trifluoromethyl)-2,6,7,8-tetrahydro-3H-
[3]benzoxocino [2,1-
clpyridin-3-y11-4-methoxy-N-([1,2,41triazolo [1,5 -alpyridin-7-yl)pentanamide
(single stereoisomer)
C1-1-4
00'
C H 3
0
Njtr-N
00 NJ,Na
CI
Diastereomer separation of 41 mg of (4S)-2-[(7R)-11-chloro-2-oxo-7-
(trifluoromethyl)-2,6,7,8-
__ tetrahydro-3H-[3]benzoxocino [2,1-c] pyridin-3 -yl] -4 -methoxy-N-
([1,2,4]triazolo [1,5 -alpyridin-7-
yl)pentanamide (mixture of two diastereomers), Example 49 gave
single stereoisomer 1 (the title compound Example 50) (chiral HPLC: Rt = 2.31
min, 99% de): 10
mg,
single stereoisomer 2 (chiral HPLC: Rt = 3.41 min, 99% de): 21 mg.
Separation method: HPLC: column: Daicel Chiralpak IC 5 um, 250 mm x 20 mm;
eluent: 60%
n-heptane / 40% 2-propanol; temperature: 40 C; flow rate: 20 ml/min; UV
detection: 220 nm.
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Analysis method: HPLC: column: Daicel Chiralpak IC 3 pm, 50 mm x 4.6 mm;
eluent: 60%
iso-heptane / 40% ethanol; temperature: 40 C; flow rate: 1 ml/min; UV
detection: 220 nm.
'H-NMR (400 MHz, DMSO-d6): 6 [ppm] = 11.09-10.88 (m, 1H), 8.87 (d, 1H), 8.39
(s, 1H), 8.23-
8.17 (m, 1H), 8.08-7.99 (m, 1H), 7.57-7.30 (m, 4H), 6.47-6.35 (m, 1H), 5.89-
5.72 (m, 1H), 4.82-4.66
(m, 1H), 4.13-3.98 / 3.82-3.74 (2m, 1H), 3.3-2.97 (m, 3H, partially
concealed), 3.19 (s, 3H), 2.85-
2.73 /2.63-2.5 (2m, 1H, partially concealed), 2.32-2.20 (m, 2H), 1.17 (d, 3H).
Additional signals of
minor rotamers were also detected.
Example 51
(45)-2 -[(7R)-11 -Chloro-2-oxo-7-(trifluoromethyl)-2,6,7, 8-tetrahydro-3H-
[3]benzoxocino [2,1-
lo clpyridin-3-y11-4-methoxy-N-(2-methyl[1,2,41triazolo[1,5-alpyridin-7-
yl)pentanamide (mixture of
two diastereomers)
CYCH3
CH3
0
H3
0 \ N
0
CI
2-Methyl[1,2,41triazolo[1,5-alpyridin-7-amine hydrochloride (24 mg, 0.13 mmol,
1.1 eq.), pyridine
(10 jil, 0.13 mmol, 1.1 eq.) and T3P (102 jil, 50% solution in ethyl acetate,
0.17 mmol, 1.5 eq.) were
added under argon atmosphere at RT to a solution of (2)-2-[(7R)-11-chloro-2-
oxo-7-
(trifluoromethyl)-2,6,7,8-tetrahydro-3H-[3]benzoxocino [2,1 -c] pyridin-3 -yll
-2,3 ,5 -trideoxy-4 -0-
methyl-L-glycero-pentonic acid (mixture of two diastereomers) (57 mg, 93%
purity, 0.12 mmol) in
tetrahydrofuran (4 m1). The reaction mixture was stirred at RT for 1 h before
all volatiles were
removed under reduced pressure. The residue was dissolved in pyridine (1 m1).
T3P (68 [11, 50%
solution in ethyl acetate, 0.12 mmol, 1.0 eq.) was added under argon
atmosphere at RT. The reaction
mixture was stirred at RT for 1 h, then all volatiles were removed under
reduced pressure. The residue
was purified by preparative HPLC (reversed phase, eluent: acetonitrile / water
gradient). Yield:
43 mg (63% of theory).
LC-MS (method 1): Rt = 0.99 min; MS (ESIpos): m/z = 590 [M+I-11+
'H-NMR (600 MHz, DM50-d6): 6 [ppm] = 11.01-10.82 (m, 1H), 8.73 (d, 1H), 8.09-
7.90 (m, 2H),
7.56-7.32 (m, 3H), 7.28-7.19 (m, 1H), 6.46-6.34 (m, 1H), 5.86-5.72 (m, 1H),
4.80-4.67 (m, 1H),
4.08-3.75 (m, 1H), 3.25-3.00 (m, 3H), 3.18 / 3.11 (2s, 3H), 2.84-2.73 /2.59-
2.46 (2m, 1H, partially
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concealed), 2.44-2.20 (m, 2H), 2.42 /2.41 (2s, 3H), 1.19-1.12 (m, 3H).
Additional signals of minor
rotamers were also detected.
Example 52
(2S,4S)-2-[(7R)-11-Chloro-2-oxo-7-(trifluoromethyl)-2,6,7,8-tetrahydro-3H-
[3]benzoxocino [2,1-
c] pyridin-3 -y1]-4-methoxy-N-(2 -methyl [1,2,4]triazolo [1,5 -a] pyridin-7-
yl)pentanamide (single
stereoisomer)
C H
C H 3
0
H3
0 N,N
0
CI
Diastereomer separation of 40 mg of (4S)-2-[(7R)-11-chloro-2-oxo-7-
(trifluoromethyl)-2,6,7,8-
tetrahydro-3H-[3]benzoxocino [2, 1-c] pyridin-3 -yl] -4 -methoxy-N-(2 -methyl
[1,2,4]triazolo [1,5 -
alpyridin-7-yl)pentanamide (mixture of two diastereomers), Example 51 gave
single stereoisomer 1 (the title compound Example 52) (chiral HPLC: R1 = 1.64
min, 99% de): 11
mg,
single stereoisomer 2 (chiral HPLC: R1 = 2.51 min, 99% de): 25 mg.
Separation method: HPLC: column: Daicel Chiralpak IC 5 p.m, 250 mm x 20 mm;
eluent: 50%
n-heptane / 50% 2-propanol; temperature: 40 C; flow rate: 20 ml/min; UV
detection: 220 nm.
Analysis method: HPLC: column: Daicel Chiralpak IC 3 pm, 50 mm x 4.6 mm;
eluent: 50% n-
heptane / 50% 2-propanol; temperature: 40 C; flow rate: 1 ml/min; UV
detection: 220 nm.
'El-NMR (400 MHz, DM50-d6): 6 [ppm] = 11.02-10.82 (m, 1H), 8.74 (d, 1H), 8.10-
7.99 (m, 2H),
7.57-7.31 (m, 3H), 7.29-7.21 (m, 1H), 6.46-6.34 (m, 1H), 5.89-5.69 (m, 1H),
4.81-4.66 (m, 1H),
.. 4.12-3.94 / 3.87-3.74 (2m, 1H), 3.3-2.98 (m, 3H, partially concealed), 3.18
(s, 3H), 2.84-2.74 / 2.63-
2.5 (2m, 1H, partially concealed), 2.42 (s, 3H), 2.31-2.20 (m, 2H), 1.19-1.12
(m, 3H). Additional
signals of minor rotamers were also detected.
Example 53
(45)-2-[(7R)-11-Chloro-2-oxo-7-(trifluoromethyl)-2,6,7,8-tetrahydro-3H-
[3]benzoxocino [2,1-
clpyridin-3-y11-4-methoxy-N-([1,2,41triazolo[1,5-alpyridin-6-yl)pentanamide
(mixture of two
diastereomers)
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(YC H3
)_C H3
0
NN
0
0
CI
[1,2,41Triazolo[1,5-alpyridin-6-amine (17 mg, 0.13 mmol, 1.1 eq.), NN-
diisopropylethylamine
(51 1.1.1, 0.29 mmol, 2.5 eq.) and a solution of HATU (53 mg, 0.14 mmol, 1.2
eq.) in NN-
dimethylformamide (1 ml) were added under argon atmosphere at RT to a solution
of (2)-2-{(7R)-
11 -chloro-2-oxo-7-(trifluoromethyl) -2,6,7, 8-tetrahydro-3H-[3]benzoxocino
[2,1 -c] pyridin-3 -yl] -
2,3,5-trideoxy-4-0-methyl-L-glycero-pentonic acid (mixture of two
diastereomers) (57 mg, 93%
purity, 0.12 mmol) in NN-dimethylformamide (3 m1). The reaction mixture was
stirred at RT for
30 min and concentrated under reduced pressure. The residue was purified by
preparative HPLC
(reversed phase, eluent: acetonitrile / water gradient). Yield: 49 mg (72% of
theory).
LC-MS (method 4): R1= 1.92 / 1.93 min; MS (ESIpos): m/z = 576 [M+H1+
'H-NMR (400 MHz, DM50-d6): 6 [ppm] = 10.95-10.75 (m, 1H), 9.42 (s, 1H), 8.46 /
8.44 (2s, 1H),
8.09-7.92 (m, 1H), 7.86 (d, 1H), 7.76-7.68 (m, 1H), 7.56-7.32 (m, 3H), 6.47-
6.35 (m, 1H), 5.89-5.74
(m, 1H), 4.80-4.65 (m, 1H), 4.14-3.74 (m, 1H), 3.3-2.98 (m, 3H, partially
concealed), 3.19 / 3.12 (2s,
3H), 2.85-2.73 / 2.64-2.5 (2m, 1H, partially concealed), 2.45-2.18 (m, 2H),
1.21-1.11 (m, 3H).
.. Additional signals of minor rotamers were also detected.
Example 54
(2S,4S)-2 -[(7R)-11 -Chloro -2-oxo -7 -(trifluoromethyl)-2,6,7,8-tetrahydro-3H-
[3]benzoxocino [2,1-
clpyridin-3-y11-4-methoxy-N-([1,2,41triazolo [1,5 -alpyridin-6-yl)pentanamide
(single stereoisomer)
(YC H3
C H 3
0
NN
0
0
CI
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Diastereomer separation of 45 mg of (4S)-2-[(7R)-11-chloro-2-oxo-7-
(trifluoromethyl)-2,6,7,8-
tetrahydro-3H-[3]benzoxocino [2,1-c] pyridin-3 -yl] -4 -methoxy-N-
(j1,2,4]triazolo [1,5 -alpyridin-6-
yl)pentanamide (mixture of two diastereomers), Example 53 gave
single stereoisomer 1 (the title compound Example 54) (chiral HPLC: Rt = 2.38
min, 99% de): 16
mg,
single stereoisomer 2 (chiral HPLC: Rt = 3.25 min, 99% de): 22 mg.
Separation method: HPLC: column: Daicel Chiralpak IE 3 um, 50 mm x 4.6 mm;
eluent: 50%
n-heptane / 50% 2-propanol; temperature: 60 C; flow rate: 25 ml/min; UV
detection: 220 nm.
Analysis method: HPLC: column: Daicel Chiralpak IE 3 pm, 50 mm x 4.6 mm;
eluent: 50% n-
heptane / 50% 2-propanol; temperature: 40 C; flow rate: 1 ml/min; UV
detection: 220 nm.
'H-NMR (400 MHz, DM50-d6): 6 [ppm] = 10.97-10.73 (m, 1H), 9.43 (s, 1H), 8.46
(s, 1H), 8.04 (br
s, 1H), 7.86 (d, 1H), 7.73 (dd, 1H), 7.57-7.33 (m, 3H), 6.48-6.35 (m, 1H),
5.90-5.73 (m, 1H), 4.82-
4.65 (m, 1H), 4.14-3.99 / 3.87-3.72 (2m, 1H), 3.3-2.97 (m, 3H, partially
concealed), 3.19 (s, 3H),
2.86-2.73 / 2.64-2.5 (2m, 1H, partially concealed), 2.32-2.15 (m, 2H), 1.16
(d, 3H). Additional
signals of minor rotamers were also detected.
Example 55
(4S)-2-[(7R)-11-Chloro-2-oxo-7-(trifluoromethyl)-2,6,7,8-tetrahydro-3H-
[3]benzoxocino [2,1-
clpyridin-3-y11-4-methoxy-N-(2-methy1-2H-benzotriazol-5-yOpentanamide (mixture
of two
diastereomers)
CH=2
0"
0
N
ci
2-Methyl-2H-benzotriazol-5-amine (19 mg, 0.13 mmol, 1.1 eq.), pyridine (11
0.13 mmol, 1.1 eq.)
and T3P (104
50% solution in ethyl acetate, 0.18 mmol, 1.5 eq.) were added under argon
atmosphere at RT to a solution of (2)-2-[(7R)-11-chloro-2-oxo-7-
(trifluoromethyl)-2,6,7,8-
tetrahydro-3H-[3]benzoxocino [2, 1-c] pyridin-3 -yl] -2,3 ,5 -trideoxy-4 -0-
methyl-L -glyce ro-pentonic
acid (mixture of two diastereomers) (57 mg, 95% purity, 0.12 mmol) in
tetrahydrofuran (4 m1). The
reaction mixture was stirred at RT for 30 min before all volatiles were
removed under reduced
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pressure. The residue was purified by preparative HPLC (reversed phase,
eluent: acetonitrile / water
gradient). Yield: 58 mg (80% of theory).
LC-MS (method 4): R1= 2.10 / 2.12 min; MS (ESIpos): m/z = 590 11M+I-11+
'H-NMR (600 MHz, DM50-d6): 6 [ppm] = 10.75-10.59 (m, 1H), 8.36-8.30 (m, 1H),
8.10-7.93 (m,
1H), 7.90-7.84 (m, 1H), 7.58-7.32 (m, 4H), 6.46-6.34 (m, 1H), 5.91-5.79 (m,
1H), 4.80-4.66 (m, 1H),
4.45 / 4.45 (2s, 3H), 4.08-3.75 (m, 1H), 3.25-2.99 (m, 3H), 3.19 / 3.12 (2s,
3H), 2.84-2.73 / 2.61-2.5
(2m, 1H, partially concealed), 2.43-2.19 (m, 2H), 1.20-1.11 (m, 3H).
Additional signals of minor
rotamers were also detected.
Example 56
(2S,4S)-2-[(7R)-11-Chloro-2-oxo-7-(trifluoromethyl)-2,6,7,8-tetrahydro-3H-
[3]benzoxocino 112, 1 -
c] pyridin-3 -y1]-4-methoxy-N-(2 -methyl-2H-benzotriazol-5 -yl)pentanamide
(single stereoisomer)
CYCH3
H 3
0
N
0
0
CI
Diastereomer separation of 55 mg of (45)-2-R7R)-11-chloro-2-oxo-7-
(trifluoromethyl)-2,6,7,8-
tetrahydro-3H-[3]benzoxocino 112, 1 -c] pyridin-3 -yl] -4 -methoxy-N-(2 -
methy1-2H-benzotriazol-5 -
yl)pentanamide (mixture of two diastereomers), Example 55 gave
single stereoisomer 1 (the title compound Example 56) (chiral HPLC: R1 = 1.18
min, 99% de): 14
mg,
single stereoisomer 2 (chiral HPLC: R1 = 1.76 min, 99% de): 29 mg.
Separation method: HPLC: column: Daicel Chiralpak IC 3 lam, 50 mm x 4.6 mm;
eluent: 50%
n-heptane / 50% 2-propanol; temperature: 60 C; flow rate: 20 ml/min; UV
detection: 220 nm.
Analysis method: HPLC: column: Daicel Chiralpak IC 3 pm, 50 mm x 4.6 mm;
eluent: 50% n-
heptane / 50% 2-propanol; temperature: 40 C; flow rate: 1 ml/min; UV
detection: 220 nm.
'H-NMR (400 MHz, DMSO-d6): 6 [ppm] = 10.77-10.60 (m, 1H), 8.33 (br s, 1H),
8.05 (br s, 1H),
7.88 (d, 1H), 7.58-7.30 (m, 4H), 6.46-6.34 (m, 1H), 5.91-5.77 (m, 1H), 4.81-
4.65 (m, 1H), 4.45 (s,
3H), 4.12-3.97 / 3.87-3.72 (2m, 1H), 3.3-2.97 (m, 3H, partially concealed),
3.19 (s, 3H), 2.86-2.73 /
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2.64-2.5 (2m, 1H, partially concealed), 2.35-2.18 (m, 2H), 1.16 (d, 3H).
Additional signals of minor
rotamers were also detected.
Example 57
(45)-2 -[(7R)-11 -Chloro-2-oxo-7-(trifluoromethyl)-2,6,7, 8-tetrahydro-3H-
[3]benzoxocino [2,1-
c] pyridin-3 -y1]-4-methoxy-N-(2 -methyl-3 -oxo-2,3 -dihydro [1,2,4]triazolo
[4,3 -a] pyridin-6-
yl)pentanamide (mixture of two diastereomers)
C 1-1,2
4C H 3
0
1-N1 0
N
N¨C H 3
\
00
CI
6-Amino-2-methyl[1,2,41triazolo[4,3-a]pyridin-3(2H)-one hydrogen chloride (26
mg, 0.13 mmol,
1.1 eq.), pyridine (28 jil, 0.35 mmol, 3.0 eq.) and T3P (102 jil, 50% solution
in ethyl acetate, 0.17
mmol, 1.5 eq.) were added under argon atmosphere at RT to a solution of (g)-2-
[(7R)-11-chloro-2-
oxo-7-(trifluoromethyl)-2,6,7,8-tetrahydro-3H- [3] benzoxocino [2,1 -c]
pyridin-3 -yll -2,3 ,5 -tride oxy-
4-0-methyl-L-glycero-pentonic acid (mixture of two diastereomers) (57 mg, 93%
purity, 0.12 mmol)
in tetrahydrofuran (4 m1). The reaction mixture was stirred at RT for 3 days
and filtered through
Celite . The filtrate removed under reduced pressure. The residue was purified
by preparative HPLC
(reversed phase, eluent: acetonitrile / water gradient). Yield: 57 mg (81% of
theory).
LC-MS (method 4): R1= 1.88 min; MS (ESIpos): m/z = 606 [M+H1+
'H-NMR (600 MHz, DM50-d6): 6 [ppm] = 10.65-10.48 (m, 1H), 8.54-8.41 (m, 1H),
8.06-7.90 (m,
1H), 7.56-7.33 (m, 3H), 7.31-7.24 (m, 2H), 6.46-6.34 (m, 1H), 5.87-5.70 (m,
1H), 4.79-4.66 (m, 1H),
4.08-3.74 (m, 1H), 3.53 / 3.53 (2s, 3H), 3.3-2.98 (m, 3H, partially
concealed), 3.18 /3.12 (2s, 3H),
2.85-2.71 / 2.60-2.5 (2m, 1H, partially concealed), 2.40-2.14 (m, 2H), 1.18-
1.10 (m, 3H). Additional
signals of minor rotamers were also detected.
Example 58
(2S,4S)-2-[(7R)-11-Chloro-2-oxo-7-(trifluoromethyl)-2,6,7,8-tetrahydro-3H-
[3]benzoxocino [2,1-
clpyridin-3-y11-4-methoxy-N-(2-methy1-3-oxo-2,3 -dihydro [1,2,4]triazolo [4,3 -
a] pyridin-6-
yl)pentanamide (single stereoisomer)
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WC H3
C H3
0
0
N
II N¨C H 3
\
00
CI
Diastereomer separation of 54 mg of (4S)-2-[(7R)-11-chloro-2-oxo-7-
(trifluoromethyl)-2,6,7,8-
tetrahydro-3H-[3]benzoxocino [2, 1-c] pyridin-3 -yl] -4 -methoxy-N-(2 -methy1-
3 -oxo -2,3 -
dihydro[1,2,41triazolo[4,3-alpyridin-6-yl)pentanamide (mixture of two
diastereomers), Example 57
gave
single stereoisomer 1 (the title compound Example 58) (chiral HPLC: Rt = 6.45
min, >99% de): 17
mg,
single stereoisomer 2 (chiral HPLC: Rt = 10.03 min, >99% de): 32 mg.
Separation method: HPLC: column: Daicel Chiralcel OX-H 5 p.m, 250 mm x 20 mm;
eluent: 60%
n-heptane / 40% ethanol; temperature: 40 C; flow rate: 20 ml/min; UV
detection: 220 nm.
Analysis method: HPLC: column: Daicel Chiralcel OX-H 5 p.m, 250 mm x 4.6 mm;
eluent: 60%
iso-hexane / 40% ethanol; temperature: 30 C; flow rate: 1 ml/min; UV
detection: 220 nm.
'H-NMR (600 MHz, DM50-d6): 6 [ppm] = 10.66-10.46 (m, 1H), 8.46 (s, 1H), 8.04-
7.94 (m, 1H),
7.56-7.32 (m, 3H), 7.31-7.25 (m, 2H), 6.46-6.33 (m, 1H), 5.83-5.69 (m, 1H),
4.79-4.65 (m, 1H),
4.10-3.98 / 3.83-3.74 (2m, 1H), 3.53 (s, 3H), 3.3-2.99 (m, 3H, partially
concealed), 3.18 (s, 3H),
2.85-2.73 / 2.63-2.5 (2m, 1H, partially concealed), 2.27-2.14 (m, 2H), 1.15
(m, 3H). Additional
signals of minor rotamers were also detected.
Example 59
4-(1(2S,4R)-2- R7R)-11-Chloro-2-oxo-7-(trifluoromethyl)-2,6,7,8 -tetrahydro-3H-
[3] benzoxocino [2,1-c] pyridin-3 -yl] -4-cyclopropy1-4-
methoxybutanoyllamino)benzamide (single
stereoisomer)
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CYC H3
FF
0
N
0
NH
2
0
1,1,3,3-Tetramethylguanidine (30
0.24 mmol, 3.0 eq.) was added under argon atmosphere at RT
to a solution of (7 R) - 11-chloro-7-(trifluoromethyl)-7,8-dihydro-3H-
[3]benzoxocinop,1-clpyridin-
2(611)-one (single stereoisomer) (27 mg, 0.08 mmol) in 2-propanol / acetone
(4:1, 1.0 m1). The
mixture was stirred at RT for 15 min, followed by addition of 4-{[(2R,4R)-2-
bromo-4-cyclopropy1-
4-methoxybutanoyll-amino}-benzamide (single stereoisomer) (31 mg, 0.09 mmol,
1.1 eq.) and of
further 2-propanol / acetone (4:1, 1.0 m1). The reaction mixture was stirred
at RT for 7 days and
concentrated under reduced pressure. The residue was purified by preparative
HPLC (reversed phase,
eluent: acetonitrile / water gradient). Yield: 28 mg (58% of theory).
LC-MS (method 16): Rt = 4.51 min; MS (ESIpos): m/z = 604 [M+I-11+
'H-NMR (600 MHz, DM50-d6): 6 [ppm] = 10.76-10.47 (m, 1H), 8.11-7.93 (m, 1H),
7.92-7.78 (m,
3H), 7.70 (d, 2H), 7.58-7.31 (m, 3H), 7.22 (br s, 1H), 6.46-6.32 (m, 1H), 5.91-
5.73 (m, 1H), 4.81-
4.65 (m, 1H), 4.11-4.00 / 3.84-3.71 (2m, 1H), 3.29-3.17 (m, 3H), 3.13-2.96 (m,
2H), 2.82-2.72 /2.63-
2.5 (2m, 2H, partially concealed), 2.44-2.20 (m, 2H), 0.90-0.77 (m, 1H), 0.63-
0.52 (m, 1H), 0.46-
0.27 (m, 2H), 0.11-0.0 (m, 1H). Additional signals of minor rotamers were also
detected.
Example 60
4- {2 -[(7R)-11 -Chloro -2 -oxo-7 -(trifluoromethyl)-2,6,7,8-tetrahydro-3H-
[3] benzoxocino [2,1 -
c] pyridin-3 -yll -4-(difluoromethoxy)butanamido -2-fluorobenzamide
(mixture of two
diastereomers)
)<H
0 F
0
N
0 1. N H 2
CI
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2-[(7R)-11 -Chloro -2 -oxo -7 -(trifluoromethyl)-2,6,7,8-tetrahydro-3H-[3]
benzoxocino 112, 1 -c] pyridin-
3-y1]-4-(difluoromethoxy)butanoic acid (mixture of two diastereomers) (63 mg,
0.13 mmol),
pyridine (12 jil, 0.14 mmol, 1.1 eq.) and T3P (114 jil, 50% solution in ethyl
acetate, 0.20 mmol, 1.5
eq.) were added under argon atmosphere at RT to a solution of 4-amino-2-
fluorobenzamide (22 mg,
0.14 mmol, 1.1 eq.) in tetrahydrofuran (4 m1). The reaction mixture was
stirred at RT for 0.5 h and
concentrated under reduced pressure. The residue was purified by preparative
HPLC (reversed phase,
eluent: acetonitrile / water gradient). Yield: 74 mg (92% of theory).
LC-MS (method 4): R1= 1.94 min; MS (ESIpos): m/z = 618 [M+I-11+
'H-NMR (600 MHz, DM50-d6): 6 [ppm] = 10.94-10.63 (m, 1H), 8.03-7.91 (m, 1H),
7.73-7.60 (m,
2H), 7.59-7.32 (m, 6H), 6.64 / 6.63 (2t, 1H), 6.47-6.35 (m, 1H), 5.81-5.58 (m,
1H), 4.78-4.63 (m,
1H), 4.17-3.73 (m, 3H), 3.28-2.96 (m, 2H), 2.84-2.72 / 2.64-2.4 (2m, 3H,
partially concealed).
Additional signals of minor rotamers were also detected.
Example 61
4-{ R25)-2 -R7R)-11 -Chloro-2-oxo -7 -(trifluoromethyl)-2,6,7,8-tetrahydro-3H-
[3] benzoxocino 112, 1 -
clpyridin-3-yll -4-(difluoromethoxy)butanoyl] amino} -2-fluorobenzamide
(single stereoisomer)
H
0 F
0 \-11
N r
00 N H 2
CI
Diastereomer separation of 72 mg of 4-{2-R7R)-11-chloro-2-oxo-7-
(trifluoromethyl)-2,6,7,8-
tetrahydro-3H-[3]benzoxocino 112, 1 -c] pyridin-3 -yl] -4 -
(difluoromethoxy)butanamido} -2-
fluorobenzamide (mixture of two diastereomers), Example 60 gave
single stereoisomer 1 (the title compound Example 61) (chiral SFC: Rt = 1.41
min, >99% de): 26
mg,
single stereoisomer 2 (chiral SFC: Rt = 2.01 min, 95% de): 23 mg.
Separation method: SFC: column: Daicel Chiralpak ID 5 um, 250 mm x 20 mm;
eluent: 82% carbon
dioxide /18% methanol; temperature: 40 C; flow rate: 80 ml/min; UV detection:
210 nm.
Analysis method: SFC: column: Daicel Chiralpak ID, 50 mm x 4.6 mm; eluent: 80%
carbon dioxide /
20% methanol; temperature: 40 C; flow rate: 3 ml/min; UV detection: 210 nm.
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'H-NMR (400 MHz, DMSO-d6): 6 [ppm] = 10.93-10.63 (m, 1H), 7.96 (br s, 1H),
7.74-7.32 (m, 8H),
6.63 (t, 1H), 6.46-6.33 (m, 1H), 5.82-5.56 (m, 1H), 4.81-4.62 (m, 1H), 4.18-
3.71 (m, 3H), 3.16-2.96
(m, 2H), 2.86-2.74 / 2.65-2.4 (2m, 3H, partially concealed). Additional
signals of minor rotamers
were also detected.
Example 62
4- { 2 -[(7R)-11 -Chloro -2 -oxo-7 -(trifluoromethyl)-2,6,7,8-tetrahydro-3H-
[3] benzoxocino [2,1 -
c] pyridin-3 -yll -4-(difluoromethoxy)butanamido}benzamide (mixture of two
diaste re omers)
)<H
0 F
0 r
N
0 *
NH
2
0
"CI
2-[(7R)-11 -Chloro -2 -oxo -7 -(trifluoromethyl)-2,6,7,8-tetrahydro-3H-
[3]benzoxocino [2,1 -clpyridin-
3-y1]-4-(difluoromethoxy)butanoic acid (mixture of two diastereomers) (77 mg,
0.16 mmol),
pyridine (14 jil, 0.18 mmol, 1.1 eq.) and T3P (140 jil, 50% solution in ethyl
acetate, 0.24 mmol, 1.5
eq.) were added under argon atmosphere at RT to a solution of 4-aminobenzamide
(24 mg, 0.18
mmol, 1.1 eq.) in tetrahydrofuran (5 m1). The reaction mixture was stirred at
RT for 0.5 h and
concentrated under reduced pressure. The residue was purified by preparative
HPLC (reversed phase,
eluent: acetonitrile / water gradient). Yield: 83 mg (84% of theory).
LC-MS (method 4): R1= 1.87 min; MS (ESIpos): m/z = 600 [M+H1+
'H-NMR (600 MHz, DM50-d6): 6 [ppm] = 10.77-10.51 (m, 1H), 8.04-7.93 (m, 1H),
7.92-7.81 (m,
3H), 7.68 (d, 2H), 7.57-7.32 (m, 3H), 7.26 (br s, 1H), 6.64 / 6.62 (2t, 1H),
6.46-6.34 (m, 1H), 5.84-
5.63 (m, 1H), 4.79-4.64 (m, 1H), 4.13-3.73 (m, 3H), 3.28-2.97 (m, 2H), 2.84-
2.72 / 2.63-2.4 (2m,
3H, partially concealed). Additional signals of minor rotamers were also
detected.
Example 63
4-{ R25)-2 -R7R)-11 -Chloro-2-oxo -7 -(trifluoromethyl)-2,6,7,8-tetrahydro-3H-
[3]benzoxocino [2,1-
c] pyridin-3 -yll -4-(difluoromethoxy)butanoyl] amino}benzamide (single
stereoisomer)
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)<H
0 F
N
0 * N H 2
0
CI
Diastereomer separation of 80 mg of 4-{2-[(7R)-11-chloro-2-oxo-7-
(trifluoromethyl)-2,6,7,8-
tetrahydro-3H-[3]benzoxocino [2, 1-c] pyridin-3 -yl] -4 -
(difluoromethoxy)butanamido}benzamide
(mixture of two diastereomers), Example 62 gave
single stereoisomer 1 (the title compound Example 63) (chiral HPLC: R1 = 1.41
min, >99% de): 20
mg,
single stereoisomer 2 (chiral HPLC: R1 = 1.74 min, >99% de): 17 mg.
Separation method: HPLC: column: Daicel Chiralpak ID 5 pm, 250 mm x 20 mm;
eluent: 70%
n-heptane / 30% ethanol; temperature: 40 C; flow rate: 20 ml/min; UV
detection: 220 nm.
Analysis method: HPLC: column: Daicel Chiralpak ID-3 3 pm, 50 mm x 4.6 mm;
eluent: 70%
n-hexane / 30% ethanol; temperature: 30 C; flow rate: 1 ml/min; UV detection:
220 nm.
'H-NMR (400 MHz, DM50-d6): 6 [ppm] = 10.77-10.48 (m, 1H), 7.97 (br s, 1H),
7.92-7.80 (m, 3H),
7.68 (d, 2H), 7.59-7.31 (m, 3H), 7.25 (br s, 1H), 6.64 (t, 1H), 6.48-6.34 (m,
1H), 5.85-5.61 (m, 1H),
4.82-4.61 (m, 1H), 4.15-3.69 (m, 3H), 3.17-2.96 (m, 2H), 2.86-2.73 / 2.65-2.4
(2m, 3H, partially
concealed). Additional signals of minor rotamers were also detected.
Example 64
4-{ [(4R)-2-[(7R)-11-Chloro-2-oxo-7-(trifluoromethyl)-2,6,7,8-tetrahydro-3H-
[3]benzoxocino [2,1-
c]pyridin-3 -y11-4-(difluoromethoxy)pentanoyll amino benzamide (mixture of two
diastereomers)
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)<H
0 F
H3
0
N
0 * N H 2
0
"CI
(2 )-2- [(7R)-11 -Chloro -2 -oxo-7 -(trifluoromethyl)-2,6,7,8-tetrahydro-3H-
[3] benzoxocino [2,1 -
clpyridin-3-y11-2,3,5-trideoxy-4-0-(difluoromethyl)-D-glycero-pentonic acid
(mixture of two
diastereomers) (99 mg, 0.20 mmol), pyridine (18 jil, 0.22 mmol, 1.1 eq.) and
T3P (175 jil, 50%
solution in ethyl acetate, 0.30 mmol, 1.5 eq.) were added under argon
atmosphere at RT to a solution
of 4-aminobenzamide (30 mg, 0.22 mmol, 1.1 eq.) in tetrahydrofuran (4 m1). The
reaction mixture
was stirred at RT for 0.5 h and concentrated under reduced pressure. The
residue was purified by
preparative HPLC (reversed phase, eluent: acetonitrile / water gradient).
Yield: 101 mg (82% of
theory).
LC-MS (method 4): R1= 1.93 min; MS (ESIpos): m/z = 614 [M+H1+
'H-NMR (400 MHz, DM50-d6): 6 [ppm] = 10.82-10.56 (m, 1H), 8.05-7.93 (m, 1H),
7.92-7.80 (m,
3H), 7.72-7.63 (m, 2H), 7.58-7.32 (m, 3H), 7.25 (br s, 1H), 6.62 / 6.59 (2t,
1H), 6.47-6.33 (m, 1H),
5.90-5.67 (m, 1H), 4.81-4.64 (m, 1H), 4.20-3.99 / 3.92-3.72 (2m, 2H), 3.16-
2.97 (m, 2H), 2.84-2.70
/ 2.65-2.5 (2m, 1H, partially concealed), 2.5-2.28 (m, 2H, partially
concealed), 1.36-1.25 (m, 3H).
Additional signals of minor rotamers were also detected.
Example 65
4-{ [(2S,4R)-2-[(7R)- 11-Chloro-2-oxo-7-(trifluoromethyl)-2,6,7,8 -tetrahydro-
3H-
[3] benzoxocino [2,1 -c] pyridin-3 -yl] -4-(difluoromethoxy)pentanoyl]
aminolbenzamide (single
stereoisomer)
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)<H
0 F
H3
0
N
0 *
NH
2
0
"CI
Diastereomer separation of 99 mg of 4-{[(4R)-2-[(7R)- 11-chloro-2-oxo-7-
(trifluoromethyl)-2,6,7,8-
tetrahydro-3H-[3]benzoxocino [2, 1-c] pyridin-3 -yl] -4 -
(difluoromethoxy)pentanoyllamino}benzamide (mixture of two diastereomers),
Example 64 gave
single stereoisomer 1 (the title compound Example 65) (chiral HPLC: Rt = 4.38
min, 99% de): 58
mg,
single stereoisomer 2 (chiral HPLC: Rt = 6.11 min, 99% de): 25 mg.
Separation method: HPLC: column: Daicel Chiralpak OZ-H 5 pm, 250 mm x 20 mm;
eluent: 55%
n-heptane / 45% ethanol; temperature: 55 C; flow rate: 15 ml/min; UV
detection: 220 nm.
Analysis method: HPLC: column: Daicel Chiralpak OZ-H 5 pm, 250 mm x 4.6 mm;
eluent: 50%
iso-hexane / 50% ethanol; temperature: 50 C; flow rate: 1 ml/min; UV
detection: 220 nm.
'H-NMR (400 MHz, DM50-d6): 6 [ppm] = 10.80-10.57 (m, 1H), 7.97 (br s, 1H),
7.91-7.80 (m, 3H),
7.68 (d, 2H), 7.58-7.32 (m, 3H), 7.24 (br s, 1H), 6.59 (t, 1H), 6.47-6.33 (m,
1H), 5.86-5.68 (m, 1H),
4.81-4.63 (m, 1H), 4.23-3.99 / 3.85-3.72 (2m, 2H), 3.17-2.97 (m, 2H), 2.83-
2.73 /2.65-2.5 (2m, 1H,
partially concealed), 2.5-2.29 (m, 2H, partially concealed), 1.30 (d, 3H).
Additional signals of minor
rotamers were also detected.
Example 66
4-{ [(45)-21 (7 R) - 11 -Chloro-2-oxo -7-(trifluoromethyl)-2,6,7,8-tetrahydro-
3H-[3]benzoxocino [2,1-
c]pyridin-3 -y11-4-(difluoromethoxy)pentanoyll amino benzamide (mixture of two
diastereomers)
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)<H
0 F
H3
0
N
00 * N H 2
0
CI
(2 )-2- [(7R)-11 -Chloro -2 -oxo-7 -(trifluoromethyl)-2,6,7,8-tetrahydro-3H-
[3] benzoxocino [2,1 -
clpyridin-3-y11-2,3,5-trideoxy-4-0-(difluoromethyl)-L-glycero-pentonic acid
(mixture of two
diastereomers) (93 mg, 89% purity, 0.17 mmol), pyridine (15 jil, 0.18 mmol,
1.1 eq.) and T3P (146
50% solution in ethyl acetate, 0.25 mmol, 1.5 eq.) were added under argon
atmosphere at RT to
a solution of 4-aminobenzamide (25 mg, 0.18 mmol, 1.1 eq.) in tetrahydrofuran
(3.6 m1). The
reaction mixture was stirred at RT for 0.5 h and concentrated under reduced
pressure. The residue
was purified by preparative HPLC (reversed phase, eluent: acetonitrile / water
gradient). Yield: 89
mg (87% of theory).
LC-MS (method 4): R1= 1.97 min; MS (ESIneg): m/z = 612 EM-H1-
'H-NMR (600 MHz, DM50-d6): 6 [ppm] = 10.81-10.53 (m, 1H), 8.07-7.94 (m, 1H),
7.92-7.80 (m,
3H), 7.72-7.64 (m, 2H), 7.57-7.31 (m, 3H), 7.26 (br s, 1H), 6.64 / 6.59 (2t,
1H), 6.45-6.35 (m, 1H),
5.88-5.70 (m, 1H), 4.80-4.65 (m, 1H), 4.25-3.73 (m, 2H), 3.13-2.96 (m, 2H),
2.83-2.72 / 2.64-2.5
(2m, 1H), 2.5-2.30 (m, 2H, partially concealed), 1.36-1.25 (m, 3H). Additional
signals of minor
rotamers were also detected.
Example 67
4-{ [(2S,45)-2- [(7R)-11 -Chl oro-2-oxo-7-(trifluoromethyl)-2,6,7,8 -
tetrahydro-3H-
[3] benzoxocino [2,1 -c] pyridin-3 -yl] -4-(difluoromethoxy)pentanoyl]
aminolbenzamide (single
stereoisomer)
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)<H
0 F
H3
0
N
L.)0 * N H 2
0
CI
Diastereomer separation of 89 mg of 4-{[(4S)-2-[(7R)-11-chloro-2-oxo-7-
(trifluoromethyl)-2,6,7,8-
tetrahydro-3H-[3]benzoxocino [2, 1-c] pyridin-3 -yl] -4 -
(difluoromethoxy)pentanoyllamino}benzamide (mixture of two diastereomers),
Example 66 gave
single stereoisomer 1 (the title compound Example 67) (chiral HPLC: R1 = 4.18
min, 99% de): 29
mg,
single stereoisomer 2 (chiral HPLC: R1 = 4.85 min, 99% de): 44 mg.
Separation method: HPLC: column: Daicel Chiralpak ID 5 lam, 250 mm x 20 mm;
eluent: 50%
n-heptane / 50% ethanol; temperature: 30 C; flow rate: 15 ml/min; UV
detection: 220 nm.
Analysis method: HPLC: column: Daicel Chiralpak ID 5 lam, 250 mm x 4.6 mm;
eluent: 50%
iso-hexane / 50% ethanol; temperature: 30 C; flow rate: 1 ml/min; UV
detection: 220 nm.
'H-NMR (400 MHz, DM50-d6): 6 [ppm] = 10.75-10.50 (m, 1H), 7.98 (br s, 1H),
7.92-7.80 (m, 3H),
7.67 (d, 2H), 7.57-7.31 (m, 3H), 7.25 (br s, 1H), 6.64 (t, 1H), 6.47-6.35 (m,
1H), 5.89-5.70 (m, 1H),
4.80-4.64 (m, 1H), 4.27-4.15 (m, 1H), 4.11-3.98 / 3.84-3.71 (2m, 1H), 3.16-
2.96 (m, 2H), 2.85-2.72
/ 2.64-2.5 (2m, 1H), 2.5-2.30 (m, 2H, partially concealed), 1.31 (d, 3H).
Additional signals of minor
rotamers were also detected.
Example 68
4- {2 -R7R)-11-Chloro -2 -oxo-7-(trifluoromethyl)-2,6,7,8-tetrahydro-3H- [3]
benzoxocino [2,1-
c] pyridin-3 -yll -4-(trifluoromethoxy)butanamido}-2-fluorobenzamide
(mixture of two
diastereomers)
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)<F
0 F
F OF
00 *
NH
2
0
CI
4-Amino-2-fluorobenzamide (12 mg, 76 umol, 1.1 eq.), /V,N-
diisopropylethylamine (30
170 umol, 2.5 eq.) and a solution of HATU (34 mg, 89 umol, 1.3 eq.) in NN-
dimethylformamide
(1.0 ml) were added under argon atmosphere at RT to a solution of 2-[(7R)-11-
chloro-2-oxo-7-
(trifluoromethyl)-2,6,7,8-tetrahydro-3H-[3]benzoxocino [2,1 -c] pyridin-3 -yll
-4 -
(trifluoromethoxy)butanoic acid (mixture of two diastereomers) (35 mg, 69
umol) in NN-
dimethylformamide (3.0 m1). The reaction mixture was stirred at RT for 1.5 h,
followed by addition
of further 4-amino-2-fluorobenzamide (3 mg, 21 umol, 0.3 eq.), NN-
diisopropylethylamine (6 IA,
34 umol, 0.5 eq.) and HATU (13 mg, 34 umol, 0.5 eq.). The reaction mixture was
stirred at RT
overnight and concentrated under reduced pressure. The residue was purified by
preparative HPLC
(reversed phase, eluent: acetonitrile / water gradient). Yield: 35 mg (80% of
theory).
LC-MS (method 1): R1= 1.08 min; MS (ESIpos): m/z = 636 [M+H1+
'H-NMR (600 MHz, DM50-d6): 6 [ppm] = 10.94-10.86 /10.65-10.59 (2m, 1H), 8.01
(d, 1H), 7.72-
7.60 (m, 2H), 7.60-7.46 (m, 3H), 7.46-7.33 (m, 3H), 6.47-6.36 (m, 1H), 5.87-
5.79 / 5.63-5.54 (2m,
1H), 4.79-4.64 (m, 1H), 4.24-4.16 (m, 1H), 4.16-4.01 / 3.94-3.73 (2m, 2H),
3.15-3.00 (m, 2H), 2.64-
2.5 (m, 3H, partially concealed). Additional signals of minor rotamers were
also detected.
Example 69
4-{ [(25)-24 (7 R) - 11 -Chloro-2-oxo -7 -(trifluoromethyl)-2,6,7,8-tetrahydro-
3H-[3]benzoxocino [2,1-
c] pyridin-3 -yll -4-(trifluoromethoxy)butanoyl] amino} -2-fluorobenzamide
(single stere oi some r)
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F
0 F
0
N
* N H 2
0
CI
Diastereomer separation of 24 mg of 4-{2-[(7R)-11-chloro-2-oxo-7-
(trifluoromethyl)-2,6,7,8-
tetrahydro-3H-[3]benzoxocino [2,1-c] pyridin-3 -yl] -4 -
(trifluoromethoxy)butanamido} -2 -
fluorobenzamide (mixture of two diastereomers), Example 68 gave
single stereoisomer 1 (the title compound Example 69) (chiral HPLC: R1 = 1.55
min, 94% de): 13
mg,
single stereoisomer 2 (chiral HPLC: R1 = 2.31 min, 99% de): 9 mg.
Separation method: HPLC: column: Daicel Chiralpak ID 5 pm, 250 mm x 20 mm;
eluent: 80%
n-heptane /20% ethanol; temperature: 40 C; flow rate: 20 ml/min; UV detection:
210 nm.
Analysis method: HPLC: column: Daicel Chiralpak ID-3 3 pm, 50 mm x 4.6 mm;
eluent: 80%
iso-hexane / 20% ethanol; temperature: 40 C; flow rate: 1 ml/min; UV
detection: 220 nm.
'H-NMR (600 MHz, DM50-d6): 6 [ppm] = 10.94-10.86 / 10.67-10.58 (m / br s, 1H),
7.99 (s, 1H),
7.73-7.60 (m, 2H), 7.60-7.46 (m, 3H), 7.46-7.23 (m, 3H), 6.48-6.36 (m, 1H),
5.86-5.78 / 5.64-5.54
(2m, 1H), 4.80-4.63 (m, 1H), 4.24-4.16 (m, 1H), 4.15-4.00 / 3.82-3.72 (2m,
2H), 3.15-2.98 (m, 2H),
2.67-2.5 (m, 3H, partially concealed). Additional signals of minor rotamers
were also detected.
Example 70
4- {2 -R7R)-11-Chloro -2 -oxo-7-(trifluoromethyl)-2,6,7,8-tetrahydro-3H- [3]
benzoxocino [2,1-
clpyridin-3-y11-4-(trifluoromethoxy)butanamidolbenzamide (mixture of two
diastereomers)
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)<F
0 F
0 ENI
N
0 *
0 NH 2
0
CI
4-Aminobenzamide (9 mg, 63 umol, 1.1 eq.), NN-diisopropylethylamine (25
140 umol, 2.5 eq.)
and a solution of HATU (28 mg, 74 umol, 1.3 eq.) in NN-dimethylformamide (1.0
ml) were added
under argon atmosphere at RT to a solution of 2-[(7R)-11-chloro-2-oxo-7-
(trifluoromethyl)-2,6,7,8-
tetrahydro-3H-[3]benzoxocino[2,1-c]pyridin-3-y1]-4-(trifluoromethoxy)butanoic
acid (mixture of
two diastereomers) (29 mg, 57 umol) in NN-dimethylformamide (2.0 m1). The
reaction mixture was
stirred at RT for 4 h, followed by addition of further 4-aminobenzamide (4 mg,
28 umol, 0.5 eq.),
NN-diisopropylethylamine (5 IA, 28 umol, 0.5 eq.) and HATU (11 mg, 28 umol,
0.5 eq.). The
reaction mixture was stirred at RT overnight and concentrated under reduced
pressure. The residue
was purified by preparative HPLC (reversed phase, eluent: acetonitrile / water
gradient). Yield: 20
mg (56% of theory).
LC-MS (method 3): Rt = 3.46 min; MS (ESIpos): m/z = 618 [M+H1+
'H-NMR (500 MHz, DM50-d6): 6 [ppm] = 10.76-10.70 /10.54-10.47 (2m, 1H), 8.01
(d, 1H), 7.91-
7.81 (m, 3H), 7.67 (d, 2H), 7.56-7.46 (m, 1H), 7.46-7.32 (m, 2H), 7.30-7.20
(m, 1H), 6.47-6.36 (m,
1H), 5.90-5.79 / 5.70-5.60 (2m, 1H), 4.80-4.65 (m, 1H), 4.23-4.15 (m, 1H),
4.15-3.96 / 3.94-3.71
(2m, 2H), 3.13-2.98 (m, 2H), 2.72-2.52 (m, 3H, partially concealed).
Additional signals of minor
rotamers were also detected.
Example 71
4-{ [2 - R7R)-11-Chloro-2-oxo-7-(trifluoromethyl)-2,6,7,8-tetrahydro-3H-
[3]benzoxocino [2,1-
clpyridin-3-yll -4-(2,2-difluoroethoxy)butanoyl] amino}benzamide (mixture of
two diastereomers)
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F.,. F
N
00
101 N H 2
0
Ci
2-[(7R)-11 -Chloro -2 -oxo -7 -(trifluoromethyl)-2,6,7,8-tetrahydro-3H-
[3]benzoxocino [2,1-c] pyridin-
3-y1]-4-(2,2-difluoroethoxy)butanoic acid (mixture of two diastereomers) (85
mg, 0.17 mmol),
pyridine (15 [11, 0.19 mmol, 1.1 eq.) and T3P (150 [11, 50% solution in ethyl
acetate, 0.26 mmol, 1.5
eq.) were added under argon atmosphere at RT to a solution of 4-aminobenzamide
(26 mg, 0.19
mmol, 1.1 eq.) in tetrahydrofuran (5 m1). The reaction mixture was stirred at
RT for 2.5 h before
water was added. The forming precipitate was filtered, washed with water and
dried in vacuo . Yield:
91 mg (86% of theory).
LC-MS (method 3): Rt = 3.37 min; MS (ESIpos): m/z = 614 [M+H1+
'H-NMR (500 MHz, DM50-d6): 6 [ppm] = 10.74-10.55 (m, 1H), 8.03-7.93 (m, 1H),
7.91-7.79 (m,
3H), 7.68 (d, 2H), 7.56-7.32 (m, 3H), 7.26 (br s, 1H), 6.45-6.32 (m, 1H), 6.21-
5.90 (m, 1H), 5.80-
5.67 (m, 1H), 4.79-4.64 (m, 1H), 4.12-3.46 (m, 5H), 3.15-2.95 (m, 2H), 2.87-
2.71 /2.65-2.5 (m, 1H,
partially concealed), 2.48-2.32 (m, 2H). Additional signals of minor rotamers
were also detected.
Example 72
4-{ R25)-2 -R7R)-11 -Chloro-2-oxo -7 -(trifluoromethyl)-2,6,7,8-tetrahydro-3H-
[3]benzoxocino [2,1-
c] pyridin-3 -yll -4-(2,2-difluoroethoxy)butanoyl] amino}benzamide (single
stereoisomer)
F F
0
N
00 10 N H 2
= 0
Ci
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Diastereomer separation of 89 mg of 4-{ [24 (7 R) - 11-chloro-2-oxo-7-
(trifluoromethyl)-2,6,7,8-
tetrahydro-3H-[3]benzoxocino [2, 1-c] pyridin-3 -yl]
difluoroethoxy)butanoyllamino}benzamide (mixture of two diastereomers),
Example 71 gave
single stereoisomer 1 (chiral HPLC: Rt = 6.40 min, 99% de): 36 mg,
single stereoisomer 2 (the title compound Example 72) (chiral HPLC: Rt = 8.95
min, 99% de): 35
mg.
Separation method: HPLC: column: Daicel Chiralpak IG 5 um, 250 mm x 20 mm;
eluent: 40%
n-heptane / 60% ethanol; temperature: 60 C; flow rate: 15 ml/min; UV
detection: 220 nm.
Analysis method: HPLC: column: Daicel Chiralpak IG 5 um, 250 mm x 4.6 mm;
eluent: 50%
iso-hexane / 50% ethanol; temperature: 60 C; flow rate: 1 ml/min; UV
detection: 220 nm.
'H-NMR (400 MHz, DM50-d6): 6 [ppm] = 10.73-10.52 (m, 1H), 7.98 (br s, 1H),
7.91-7.80 (m, 3H),
7.68 (d, 2H), 7.56-7.33 (m, 3H), 7.25 (br s, 1H), 6.46-6.33 (m, 1H), 6.07 (tt,
1H), 5.80-5.67 (m, 1H),
4.80-4.64 (m, 1H), 4.13-3.43 (m, 5H), 3.16-2.95 (m, 2H), 2.87-2.76 / 2.64-2.5
(2m, 1H, partially
concealed), 2.48-2.34 (m, 2H). Additional signals of minor rotamers were also
detected.
Example 73
4-( {2- R7R)-11-Chloro-2-oxo-7-(trifluoromethyl)-2,6,7,8-tetrahydro-3H-
[3]benzoxocino [2,1-
clpyridin-3-y11-4-Rpropan-2-yDoxylbutanoyll amino)-2-fluorobenzamide (mixture
of two
diastereomers)
C H 3
0 )C H 3
0
1
-
N N
0 I N H 2
0
CI
4-Amino-2-fluorobenzamide (27 mg, 0.17 mmol, 1.1 eq.), NN-
diisopropylethylamine (68 IA,
0.39 mmol, 2.5 eq.) and a solution of HATU (78 mg, 0.20 mmol, 1.3 eq.) in NN-
dimethylformamide
(1 ml) were added under argon atmosphere at RT to a solution of 2-[(7R)-11-
chloro-2-oxo-7-
(trifluoromethyl)-2,6,7,8-tetrahydro-3H-[3]benzoxocino [2,1-c] pyridin-3 -yll -
4 -Rpropan-2-
yl)oxylbutanoic acid (mixture of two diastereomers) (81 mg, 92% purity, 0.16
mmol) in NN-
dimethylformamide (4 m1). The reaction mixture was stirred at RT for 3 h and
concentrated under
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reduced pressure. The residue was purified by preparative HPLC (reversed
phase, eluent: acetonitrile
/ water gradient). Yield: 64 mg (66% of theory).
LC-MS (method 4): Rt = 2.05 min; MS (ESIpos): m/z = 610 [M+I-11+
'H-NMR (500 MHz, DM50-d6): 6 [ppm] = 10.92-10.67 (m, 1H), 8.04-7.93 (m, 1H),
7.72-7.61 (m,
2H), 7.60-7.30 (m, 6H), 6.46-6.33 (m, 1H), 5.79-5.64 (m, 1H), 4.79-4.64 (m,
1H), 4.10-3.94 / 3.92-
3.72 (2m, 1H), 3.53-3.38 (m, 2H), 3.15-2.98 (m, 2H), 2.81-2.69 / 2.63-2.4 (2m,
1H, partially
concealed), 2.43-2.25 (m, 2H), 1.05-0.90 (m, 6H). Additional signals of minor
rotamers were also
detected.
Example 74
4-( (25)-2 -[(7R)-11 -Chloro-2-oxo -7-(trifluoromethyl)-2,6,7,8-tetrahydro-3H-
[3]benzoxocino [2,1-
c] pyridin-3 -y11-4-[(propan-2-yl)oxy] butanoyl} amino)-2-fluorobenzamide
(single stereoisomer)
C H 3
0 )C H 3
0
N
00 N H 2
CI
Diastereomer separation of 60 mg of 4-({2-[(7R)-11-chloro-2-oxo-7-
(trifluoromethyl)-2,6,7,8-
tetrahydro-3H-[3]benzoxocino 112, 1 -c] pyridin-3 -yl] -4 -Rpropan-2-
yl)oxylbutanoyllamino)-2 -
fluorobenzamide (mixture of two diastereomers), Example 73 gave
single stereoisomer 1 (the title compound Example 74) (chiral HPLC: R1 = 1.29
min, >99% de): 16
mg,
single stereoisomer 2 (chiral HPLC: R1 = 2.03 min, >99% de): 14 mg.
Separation method: HPLC: column: Daicel Chiralpak ID 5 lam, 250 mm x 20 mm;
eluent: 70%
n-heptane / 30% ethanol; temperature: 40 C; flow rate: 20 ml/min; UV
detection: 220 nm.
Analysis method: HPLC: column: Daicel Chiralpak ID 3 pm, 50 mm x 4.6 mm;
eluent: 70% n-
heptane / 30% ethanol; temperature: 40 C; flow rate: 1 ml/min; UV detection:
220 nm.
'H-NMR (400 MHz, DM50-d6): 6 [ppm] = 10.87-10.67 (m, 1H), 7.97 (s, 1H), 7.75-
7.60 (m, 2H),
7.58-7.31 (m, 6H), 6.47-6.32 (m, 1H), 5.81-5.63 (m, 1H), 4.81-4.62 (m, 1H),
4.13-3.99 / 3.83-3.70
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(2m, 1H), 3.55-3.39 (m, 2H), 3.16-2.97 (m, 2H), 2.82-2.69 / 2.63-2.4 (2m, 1H,
partially concealed),
2.43-2.25 (m, 2H), 1.05-0.93 (m, 6H). Additional signals of minor rotamers
were also detected.
Example 75
4-(1(2-R7R)-11 -Chloro-2-oxo-7-(trifluoromethyl)-2,6,7, 8-tetrahydro-3H-
[3]benzoxocino [2,1-
clpyridin-3-y11-4-[(propan-2-yl)oxylbutanoyl}amino)benzamide (mixture of two
diastereomers)
C H 3
0 )C H 3
0
ENI
N
00 N H 2
0
CI
4-Aminobenzamide (24 mg, 0.17 mmol, 1.1 eq.), NN-diisopropylethylamine (68
0.39 mmol, 2.5
eq.) and a solution of HATU (78 mg, 0.20 mmol, 1.3 eq.) in NN-
dimethylformamide (1 ml) were
added under argon atmosphere at RT to a solution of 2-[(7R)-11-chloro-2-oxo-7-
(trifluoromethyl)-
2,6,7,8-tetrahydro -3H-[3]benzoxocino [2,1 -c] pyridin-3 -yl] -4- Rpropan-2 -
yl)oxy] butanoic acid
(mixture of two diastereomers) (81 mg, 92% purity, 0.16 mmol) in NN-
dimethylformamide (4 m1).
The reaction mixture was stirred at RT for 2 h and concentrated under reduced
pressure. The residue
was crystallized with water, filtered, washed with water and dissolved in
acetonitrile. This solution
was used in purification by preparative HPLC (reversed phase, eluent:
acetonitrile / water gradient).
Yield: 55 mg (59% of theory).
LC-MS (method 4): R1= 1.98 min; MS (ESIpos): m/z = 592 [M+H1+
'H-NMR (500 MHz, DM50-c/6): 6 [ppm] = 10.75-10.55 (m, 1H), 8.05-7.94 (m, 1H),
7.88 (br s, 1H),
7.85 (d, 2H), 7.70 (d, 2H), 7.57-7.32 (m, 3H), 7.27 (br s, 1H), 6.45-6.33 (m,
1H), 5.82-5.69 (m, 1H),
4.80-4.65 (m, 1H), 4.10-3.95 / 3.92-3.72 (2m, 1H), 3.54-3.39 (m, 2H), 3.16-
2.97 (m, 2H), 2.80-2.70
/ 2.62-2.45 (2m, 1H, partially concealed), 2.44-2.26 (m, 2H), 1.05-0.91 (m,
6H). Additional signals
of minor rotamers were also detected.
Example 76
4-(1(25)-2-R7R)- 11 -Chloro-2-oxo -7-(trifluoromethyl)-2,6,7,8-tetrahydro-3H-
[3]benzoxocino [2,1-
clpyridin-3-y11-4-Rpropan-2-yDoxylbutanoyll amino)benzamide (single
stereoisomer)
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C H 3
0 )C H 3
0
ENI
N
00 N H 2
0
CI
Diastereomer separation of 55 mg of 4-({(2-[(7R)-11-chloro-2-oxo-7-
(trifluoromethyl)-2,6,7,8-
tetrahydro-3H-[3]benzoxocino [2, 1 -c] pyridin-3 -yl] -4 -Rpropan-2-
yl)oxylbutanoyl amino)benzamide (mixture of two diastereomers), Example 75
gave
single stereoisomer 1 (the title compound Example 76) (chiral SFC: R1 = 4.13
min, >99% de): 19
mg,
single stereoisomer 2 (chiral SFC: R1= 6.13 min, >99% de): 15 mg.
Separation method: SFC: column: Daicel Chiralpak IE 5 p.m, 250 mm x 20 mm;
eluent: 74% carbon
dioxide /26% methanol; temperature: 40 C; flow rate: 80 ml/min; UV detection:
210 nm.
Analysis method: SFC: column: Daicel Chiralpak IE, 50 mm x 4.6 mm; eluent: 70%
carbon dioxide /
30% methanol; temperature: 40 C; flow rate: 3 ml/min; UV detection: 220 nm.
'H-NMR (400 MHz, DM50-d6): 6 [ppm] = 10.72-10.53 (m, 1H), 7.97 (s, 1H), 7.92-
7.79 (m, 3H),
7.69 (d, 2H), 7.57-7.33 (m, 3H), 7.24 (br s, 1H), 6.46-6.33 (m, 1H), 5.80-5.68
(m, 1H), 4.81-4.64 (m,
1H), 4.12-3.99 / 3.84-3.71 (2m, 1H), 3.54-3.39 (m, 2H), 3.15-2.97 (m, 2H),
2.80-2.71 /2.64-2.5 (2m,
1H, partially concealed), 2.44-2.25 (m, 2H), 1.06-0.94 (m, 6H). Additional
signals of minor rotamers
were also detected.
Example 77
4-( {4-tert-Butoxy-2-[(7R)-11-chloro-2-oxo-7-(trifluoromethyl) -2,6,7,8-
tetrahydro-3H-
[3] benzoxocino 112,1 -c] pyridin-3 -yllbutanoyllamino)benzamide (mixture of
two diastereomers)
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H 3C
*CH3
0 C H 3
0
ENI
N
00 el N H 2
0
Ci
4-tert-Butoxy-2-{(7R)-11 -chloro-2-oxo -7 -(trifluoromethyl)-2,6,7, 8-
tetrahydro-3H-
[3]benzoxocino [2,1-c] pyridin-3 -yll butanoic acid (mixture of two
diastereomers) (53 mg, 0.11
mmol), pyridine (9 jil, 0.12 mmol, 1.1 eq.) and T3P (92 jil, 50% solution in
ethyl acetate, 0.16 mmol,
1.5 eq.) were added under argon atmosphere at RT to a solution of 4-
aminobenzamide (16 mg, 0.12
mmol, 1.1 eq.) in tetrahydrofuran (3 m1). The reaction mixture was stirred at
RT for 2.5 h before
water was added. The forming precipitate was filtered, washed with water and
dried in vacuo . Yield:
51 mg (80% of theory).
LC-MS (method 3): Rt = 3.62 min; MS (ESIpos): m/z = 606 [M+H1+
'H-NMR (600 MHz, DM50-d6): 6 [ppm] = 10.75-10.55 (m, 1H), 8.02 / 7.97 (2s,
1H), 7.86 (br s,
1H), 7.85 (d, 2H), 7.70 (d, 2H), 7.57-7.31 (m, 3H), 7.25 (br s, 1H), 6.46-6.33
(m, 1H), 5.83-5.70 (m,
1H), 4.79-4.66 (m, 1H), 4.09-3.94 / 3.91-3.72 (2m, 1H), 3.63-3.57 / 3.47-3.3
(2m, 2H, partially
concealed), 3.15-3.01 (m, 2H), 2.76-2.67 /2.62-2.5 (2m, 1H, partially
concealed), 2.42-2.25 (m, 2H),
1.04 / 1.01 (2s, 9H). Additional signals of minor rotamers were also detected.
Example 78
4-(1(25)-4 -tert-Butoxy-2-R7R)-11 -chloro-2 -oxo -7-(trifluoromethyl)-2,6,7,8-
tetrahydro -3H-
[3] benzoxocino [2,1 -c] pyridin-3 -yll butanoyllamino)benzamide (single stere
oi some r)
H 3C
*CH3
0 C H3
0
ENI
N
00 el N H
0
Ci
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Diastereomer separation of 50 mg of 4 -( {4-tert-butoxy-2 -R7R)-11-chloro-2-
oxo-7-
(trifluoromethyl)-2,6,7,8-tetrahydro-3H-[3]benzoxocino [2,1-c] pyridin-3 -
yllbutanoyl}amino)benzamide (mixture of two diastereomers), Example 77 gave
single stereoisomer 1 (the title compound Example 78) (chiral HPLC: Rt = 1.18
min, >99% de): 18
mg,
single stereoisomer 2 (chiral HPLC: Rt = 1.71 min, 99% de): 17 mg.
Separation method: HPLC: column: Daicel Chiralpak ID 5 p.m, 250 mm x 20 mm;
eluent: 70%
n-heptane / 30% ethanol; temperature: 40 C; flow rate: 25 ml/min; UV
detection: 220 nm.
Analysis method: HPLC: column: Daicel Chiralpak ID 3 pm, 50 mm x 4.6 mm;
eluent: 70% n-
hexane / 30% ethanol; temperature: 30 C; flow rate: 1 ml/min; UV detection:
220 nm.
'El-NMR (400 MHz, DM50-d6): 6 [ppm] = 10.70-10.55 (m, 1H), 7.97 (s, 1H), 7.89-
7.81 (m, 3H),
7.70 (d, 2H), 7.57-7.31 (m, 3H), 7.24 (br s, 1H), 6.45-6.34 (m, 1H), 5.83-5.70
(m, 1H), 4.80-4.66 (m,
1H), 4.09-3.96 / 3.85-3.72 (2m, 1H), 3.48-3.3 (m, 2H, partially concealed),
3.15-3.00 (m, 2H), 2.76-
2.67 / 2.62-2.5 (2m, 1H, partially concealed), 2.40-2.26 (m, 2H), 1.04 (s,
9H). Additional signals of
minor rotamers were also detected.
Example 79
4-{ [2 - R7R)-11-Chloro-2-oxo-7-(trifluoromethyl)-2,6,7, 8-tetrahydro-3H-
[3]benzoxocino [2,1-
c] pyridin-3 -yll -4-(cyclopropyloxy)butanoyl] amino}benzamide (mixture of two
diastereomers)
o/A
0
EN1
N
0 N H 2
0
0
CI
2-[(7R)-11 -Chloro -2 -oxo -7-(trifluoromethyl)-2,6,7,8-tetrahydro-3H-
[3]benzoxocino [2,1-clpyridin-
3-y11-4-(cyclopropyloxy)butanoic acid (mixture of two diastereomers) (66 mg,
0.14 mmol), pyridine
(12 jil, 0.15 mmol, 1.1 eq.) and T3P (123 jil, 50% solution in ethyl acetate,
0.21 mmol, 1.5 eq.) were
added under argon atmosphere at RT to a solution of 4-aminobenzamide (21 mg,
0.15 mmol, 1.1 eq.)
in tetrahydrofuran (3 m1). The reaction mixture was stirred at RT for 2.5 h
before water was added.
The forming precipitate was filtered, washed with water and dried in vacuo.
Yield: 33 mg (40% of
theory).
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The combined filtrates were extracted with ethyl acetate. The combined organic
phases were washed
with brine, dried over anhydrous sodium sulfate, filtered and concentrated
under reduced pressure.
Yield: 24 mg (28% of theory).
24 mg-batch: LC-MS (method 4): Rt = 1.93 min; MS (ESIpos): m/z = 590 [M-411+
33 mg-batch: 'H-NMR (500 MHz, DM50-d6): 6 [ppm] = 10.74-10.57 (m, 1H), 8.04-
7.93 (m, 1H),
7.87 (br s, 1H), 7.85 (d, 2H), 7.69 (d, 2H), 7.57-7.32 (m, 3H), 7.26 (br s,
1H), 6.46-6.33 (m, 1H),
5.80-5.66 (m, 1H), 4.80-4.66 (m, 1H), 4.10-3.95 / 3.93-3.72 (2m, 1H), 3.57-3.3
(m, 2H, partially
concealed), 3.27-3.20 (m, 1H), 3.14-2.99 (m, 2H), 2.82-2.71 /2.63-2.5 (2m, 1H,
partially concealed),
2.44-2.25 (m, 2H), 0.45-0.23 (m, 4H). Additional signals of minor rotamers
were also detected.
Example 80
4-{ R2S)-2-R7R)-11-Chloro-2-oxo-7-(trifluoromethyl)-2,6,7,8-tetrahydro-3H-
[3]benzoxocino 112, 1-
clpyridin-3-y11-4-(cyclopropyloxy)butanoyllamino}benzamide (single
stereoisomer)
0
ENI
N
00 10 N H 2
0
Ci
Diastereomer separation of 55 mg of 4-{ [2-R7R)-11-chloro-2-oxo-7-
(trifluoromethyl)-2,6,7,8-
tetrahydro-3H-[3]benzoxocino 112, 1-c] pyridin-3 -yl] -4 -
(cyclopropyloxy)butanoyl] amino}benzamide
(mixture of two diastereomers), Example 79 gave
single stereoisomer 1 (chiral HPLC: Rt = 7.13 min, 99% de): 24 mg,
single stereoisomer 2 (the title compound Example 80) (chiral HPLC: Rt = 11.69
min, 99% de): 26
mg.
Separation method: HPLC: column: Daicel Chiralpak IG 5 lam, 250 mm x 20 mm;
eluent: 40%
n-heptane / 60% ethanol; temperature: 60 C; flow rate: 15 ml/min; UV
detection: 220 nm.
Analysis method: HPLC: column: Daicel Chiralpak IG 5 lam, 250 mm x 4.6 mm;
eluent: 50%
iso-hexane / 50% ethanol; temperature: 60 C; flow rate: 1 ml/min; UV
detection: 220 nm.
'H-NMR (400 MHz, DM50-d6): 6 [ppm] = 10.72-10.55 (m, 1H), 7.97 (br s, 1H),
7.91-7.80 (m, 3H),
7.68 (d, 2H), 7.57-7.33 (m, 3H), 7.24 (br s, 1H), 6.46-6.33 (m, 1H), 5.80-5.66
(m, 1H), 4.81-4.65 (m,
1H), 4.10-3.99 / 3.83-3.72 (2m, 1H), 3.58-3.3 (m, 2H, partially concealed),
3.27-3.20 (m, 1H), 3.14-
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2.99 (m, 2H), 2.82-2.74 / 2.63-2.5 (2m, 1H, partially concealed), 2.44-2.25
(m, 2H), 0.46-0.29 (m,
4H). Additional signals of minor rotamers were also detected.
Example 81
4-( {2-[(7R)-11-Chloro-2-oxo-7-(trifluoromethyl)-2,6,7,8-tetrahydro-3H-
[3]benzoxocino [2,1-
clpyridin-3-yll -4-[(1-methylcyclopropyl)oxy] butanoyl}amino)-2-
fluorobenzamide (mixture of two
diastereomers)
C H 3
0 )7
0
ENI
N
0 10 N H 2
= 0
C I
4-Amino-2-fluorobenzamide (15 mg, 0.10 mmol, 1.1 eq.), /V,N-
diisopropylethylamine (38
0.22 mmol, 2.5 eq.) and a solution of HATU (43 mg, 0.11 mmol, 1.3 eq.) in /V,N-
dimethylformamide
(1 ml) were added under argon atmosphere at RT to a solution of 2-[(7R)-11-
chloro-2-oxo-7-
(trifluoromethyl)-2,6,7,8-tetrahydro-3H-[3]benzoxocino [2,1-c] pyridin-3 -yll -
4 -[(1 -
methylcyclopropy1)-oxylbutanoic acid (mixture of two diastereomers) (45 mg,
93% purity, 0.09
mmol) in NN-dimethylformamide (2 m1). After stirring at RT for 4.5 h,
additionally 4-amino-2-
fluorobenzamide (7 mg, 0.04 mmol, 0.5 eq.) and HATU (16 mg, 0.04 mmol, 0.5
eq.) were added.
The reaction mixture was stirred for 2 days and concentrated under reduced
pressure. The residue
was purified by preparative HPLC (reversed phase, eluent: acetonitrile / water
gradient). Yield: 10
mg (19% of theory).
LC-MS (method 4): R1= 2.10 min; MS (ESIpos): m/z = 622 [M+H1+
'H-NMR (600 MHz, DM50-c/6): 6 [ppm] = 10.90-10.70 (m, 1H), 8.01 /7.96 (2s,
1H), 7.72-7.61 (m,
2H), 7.59-7.47 (m, 3H), 7.46-7.33 (m, 3H), 6.46-6.33 (m, 1H), 5.76-5.63 (m,
1H), 4.78-4.66 (m, 1H),
4.10-3.94 / 3.91-3.73 (2m, 1H), 3.57-3.45 (m, 1H), 3.15-3.01 (m, 2H), 2.80-
2.70 / 2.62-2.5 (2m, 1H,
partially concealed), 2.41-2.27 (m, 2H), 1.27-1.16 (m, 3H), 0.64-0.56 (m, 1H),
0.54-0.38 (m, 1H),
0.35-0.22 (m, 2H). One proton is concealed. Additional signals of minor
rotamers were also detected.
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Example 82
4-( {2-[(7R)-11-Chloro-2-oxo-7-(trifluoromethyl)-2,6,7,8-tetrahydro-3H-
[3]benzoxocino [2,1-
c] pyridin-3 -yll -4- R1-methylcyclopropyl)oxy] butanoyllamino)benzamide
(mixture of two
diastereomers)
C H 3
0
0
\
N
0 N H2
0
= 0
CI
4-Aminobenzamide (39 mg, 0.29 mmol, 1.1 eq.), NN-diisopropylethylamine (113
0.65 mmol,
2.5 eq.) and a solution of HATU (129 mg, 0.34 mmol, 1.3 eq.) in NN-
dimethylformamide (1 ml)
were added under argon atmosphere at RT to a solution of 2-[(7R)-11-chloro-2-
oxo-7-
(trifluoromethyl)-2,6,7,8-tetrahydro-3H-[3]benzoxocino [2,1-c] pyridin-3 -yll -
4 -[(1-
methylcyclopropyl)oxylbutanoic acid (mixture of two diastereomers) (136 mg,
93% purity, 0.26
mmol) in NN-dimethylformamide (4 m1). The reaction mixture was stirred at RT
for 3 h and
concentrated under reduced pressure at 50 C water bath temperature. The
residue was crystallized
with water, filtered, washed with water and dried in vacuo. This residue was
purified by column
chromatography (silica gel, eluent: dichloromethane / methanol gradient).
Yield: two batches
isolated, 83 mg and 48 mg (82% of theory for both batches together).
both batches: LC-MS (method 4): Rt = 2.03 min; MS (ESIpos): m/z = 604 [M+H1+
48mg-batch: 'H-NMR (500 MHz, DM50-d6): 6 [ppm] = 10.72-10.56 (m, 1H), 8.01 /
7.96 (2s, 1H),
7.86 (br s, 1H), 7.84 (d, 2H), 7.69 (d, 2H), 7.56-7.33 (m, 3H), 7.25 (br s,
1H), 6.46-6.34 (m, 1H),
5.78-5.66 (m, 1H), 4.80-4.66 (m, 1H), 4.09-3.96 / 3.92-3.72 (2m, 1H), 3.66-
3.45 (m, 1H), 3.3-3.20
(m, 1H, partially concealed), 3.17-3.01 (m, 2H), 2.78-2.71 /2.60-2.5 (2m, 1H,
partially concealed),
2.42-2.25 (m, 2H), 1.29-1.18 (m, 3H), 0.66-0.56 (m, 1H), 0.54-0.39 (m, 1H),
0.36-0.22 (m, 2H).
Additional signals of minor rotamers were also detected.
Example 83
4-(1(25)-2-R7R)- 11 -Chloro-2-oxo -7-(trifluoromethyl)-2,6,7,8-tetrahydro-3H-
[3]benzoxocino [2,1-
clpyridin-3-yll -4-[(1-methylcyclopropyl)oxy] butanoyl}amino)benzamide (single
stereoisomer)
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C H 3
0 )7
0
E
N NI
0 N H 2
0
Ci
Diastereomer separation of 122 mg of 4-({2-[(7R)-11-chloro-2-oxo-7-
(trifluoromethyl)-2,6,7,8-
tetrahydro-3H-[3]benzoxocino [2, 1-c] pyridin-3 -yl] -4 -R1-
methylcyclopropyl)oxylbutanoyllamino)-
benzamide (mixture of two diastereomers), Example 82 gave
single stereoisomer 1 (the title compound Example 83) (chiral HPLC: Rt = 2.49
min, 99% de): 75
mg,
single stereoisomer 2 (chiral HPLC: Rt = 3.43 min, 98% de): 45 mg.
Separation method: HPLC: column: Daicel Chiralpak OZ-H 5 pm, 250 mm x 20 mm;
eluent: 50%
n-heptane / 50% ethanol; temperature: 40 C; flow rate: 25 ml/min; UV
detection: 220 nm.
Analysis method: HPLC: column: Daicel Chiralpak OZ-H 3 pm, 50 mm x 4.6 mm;
eluent: 50%
iso-hexane / 50% ethanol; temperature: 40 C; flow rate: 1 ml/min; UV
detection: 220 nm.
'H-NMR (400 MHz, DM50-d6): 6 [ppm] = 10.71-10.55 (m, 1H), 7.96 (s, 1H), 7.90-
7.80 (m, 3H),
7.69 (d, 2H), 7.57-7.33 (m, 3H), 7.24 (br s, 1H), 6.46-6.33 (m, 1H), 5.79-5.66
(m, 1H), 4.80-4.65 (m,
1H), 4.10-4.00 / 3.83-3.72 (2m, 1H), 3.67-3.44 (m, 1H), 3.37-3.3 (m, 1H,
partially concealed), 3.18-
3.00 (m, 2H), 2.79-2.69 / 2.63-2.5 (2m, 1H, partially concealed), 2.89-2.25
(m, 2H), 1.28-1.18 (m,
3H), 0.66-0.58 (m, 1H), 0.55-0.44 (m, 1H), 0.37-0.25 (m, 2H). Additional
signals of minor rotamers
were also detected.
Example 84
4- {2 -R7R)-11-Chloro -2 -oxo-7-(trifluoromethyl)-2,6,7,8-tetrahydro-3H- [3]
benzoxocino [2,1-
clpyridin-3-y11-4-(cyclobutyloxy)butanamido}benzamide (mixture of two
diastereomers)
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o
0 H
N
0
NH
2
0
0
"CI
General Method 11 was carried out with 2-[(7R)-11-chloro-2-oxo-7-
(trifluoromethyl)-2,6,7,8-
tetrahydro-3H-[3]benzoxocino[2,1-c]pyridin-3-y1]-4-(cyclobutyloxy)butanoic
acid (mixture of two
diastereomers) (8.0 mg, 16 lama 1.0 eq.), 4-aminobenzamide (3.36 mg, 25 lama
1.5 eq.) and T3P
(39 [11, 50% solution in ethyl acetate, 66 lama 4.0 eq.) in pyridine (0.5 ml)
including the following
variations of the procedure: The reaction mixture was stirred at 50 C for 2 h,
concentrated under
reduced pressure and purified by preparative HPLC (reversed phase, eluent:
water with 0.05% formic
acid! acetonitrile 80:20 to 5:95). Yield: 2.3 mg (22% of theory).
LC-MS (method 3): Rt = 3.58 min; MS (ESIpos): m/z = 604 [M+H1+
Example 85
4-{ R25)-2 -R7R)-11 -Chloro-2-oxo -7 -(trifluoromethyl)-2,6,7,8-tetrahydro-3H-
[3] benzoxocino [2,1-
c] pyridin-3 -yll -4-(cyclobutyloxy)butanoyl] amino}benzamide (single stere oi
some r)
0 IN
N
0
NH
0
Ci
General Method 11 was carried out two times: the first time with 2-[(7R)-11-
chloro-2-oxo-7-
(trifluoromethyl)-2,6,7,8-tetrahydro-3H-[3]benzoxocino [2,1-c] pyridin-3 -yll -
4 -
(cyclobutyloxy)butanoic acid (mixture of two diastereomers) (74.0 mg, 152 lama
1.0 eq.), 4-
aminobenzamide (31.1 mg, 228 lama 1.5 eq.) and T3P (360 [11, 50% solution in
ethyl acetate, 610
lama 4.0 eq.) in pyridine (4.6 ml) and the second time with 2-[(7R)-11-chloro-
2-oxo-7-
(trifluoromethyl)-2,6,7,8-tetrahydro-3H-[3]benzoxocino [2,1-c] pyridin-3 -yll -
4-
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(cyclobutyloxy)butanoic acid (mixture of two diastereomers) (30.0 mg, 62 umol,
1.0 eq.), 4-
aminobenzamide (12.6 mg, 93 umol, 1.5 eq.) and T3P (147 IA, 50% solution in
ethyl acetate, 247
umol, 4.0 eq.) in pyridine (1.9 ml) including the following variations of the
procedure: The reaction
mixtures were stirred for 4 h at 50 C, concentrated under reduced pressure and
purified by
preparative HPLC (reversed phase, eluent: water with 0.05% formic acid /
acetonitrile 80:20 to 5:95).
The two product batches were combined and 50 mg of 4-{2-[(7R)-11-chloro-2-oxo-
7-
(trifluoromethyl)-2,6,7,8-tetrahydro-3H-[3]benzoxocino [2,1-c] pyridin-3 -yll -
4 -
(cyclobutyloxy)butanamido}benzamide (mixture of two diastereomers) were
further purified by
diastereomer separation to provide:
single stereoisomer 1 (the title compound 85) (chiral HPLC: R1= 1.00 min, 92%
de): 12.8 mg (10%
of theory),
single stereoisomer 2 (chiral HPLC: R1 = 1.03 min): 13.2 mg.
Separation method: HPLC: column: Daicel Chiralpak IF 5 um, 250 mm x 20 mm;
eluent: 40%
n-heptane / 60% ethanol; temperature: 40 C; flow rate: 20 ml/min; UV
detection: 220 nm.
Analysis method: HPLC: column: Daicel Chiraltek IC 3 um, 50 mm x 4.6 mm;
eluent: 50% n-
heptane / 50% ethanol; flow rate: 1.0 ml/min; UV detection: 220 nm.
LC-MS (method 1): R1 = 1.11 min; MS (ESIpos): m/z = 604 [M+I-11+
1H-NMR (500 MHz, DM50-d6): 6 [ppm] = 10.73 (s, 1H), 8.03 (s, 1H), 7.90-7.80
(m, 3H), 7.73-7.60
(m, 2H), 7.53 (br d, 1H), 7.47-7.33 (m, 2H), 7.24 (br s, 1H), 6.44 / 6.40 (2s,
1H), 5.84-5.78 (m, 1H),
4.77-4.69 (m, 1H), 3.86 (t, 1H), 3.27-3.14 (m, 1H), 3.14-2.99 (m, 2H), 2.48-
2.34 (m, 2H), 2.13-1.90
(m, 1H), 1.85-1.72 (m, 1H), 1.72-1.58 (m, 2H), 1.57-1.38 (m, 2H), 1.30-1.14
(m, 3H). Additional
signals of minor rotamers were also detected.
Example 86
4- {2 -R7R)-11-Chloro -2 -oxo-7 -(trifluoromethyl)-2,6,7,8-tetrahydro-3H- [3]
benzoxocino [2,1-
clpyridin-3-yll -44(1-methylcyclobutypoxylbutanamidolbenzamide (mixture of two
diastereomers)
0 r, u
0
N
0 N H 2
Ci
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General Method 11 was carried out with 2-[(7R)-11-chloro-2-oxo-7-
(trifluoromethyl)-2,6,7,8-
tetrahydro-3H-[3]benzoxocino [2,1-c] pyridin-3 -yl] -4 -R1-
methylcyclobutypoxy] butanoic acid
(mixture of two diastereomers) (44.5 mg, 89.0 umol, 1.0 eq.), 4-aminobenzamide
(18.2 mg, 134
umol, 1.5 eq.) and T3P (210 jil, 50% solution in ethyl acetate, 360 umol, 4.0
eq.) in pyridine (2.7
ml) including the following variations of the procedure: The crude mixture was
purified by
preparative HPLC (reversed phase, eluent: water with 0.05% formic acid /
acetonitrile 80:20 to 5:95).
Yield: 35 mg (64% of theory).
LC-MS (method 1): R1= 1.12 min; MS (ESIpos): m/z = 618 [M+H1+
Example 87
4-( {(2,S) -2 -R7R)- 11 -Chloro-2-oxo -7-(trifluoromethyl)-2,6,7,8-tetrahydro-
3H-[3]benzoxocino [2,1-
c] pyridin-3 -yll -4- R1-methylcyclobutypoxylbutanoyllamino)benzamide (single
stereoisomer)
0417
C H 3
0
N
0
NH
2
0
= 0
CI
Diastereomer separation of 28 mg of 4-{2-[(7R)-11-chloro-2-oxo-7-
(trifluoromethyl)-2,6,7,8-
tetrahydro-3H-[3]benzoxocino [2, 1-c] pyridin-3 -yl] -4 -{(1-
methylcyclobutypoxylbutanamido}benzamide (mixture of two diastereomers),
Example 86 gave
single stereoisomer 1 (the title compound 87) (chiral HPLC: R1 = 5.44 min, 99%
de): 10.6 mg (19%
of theory),
single stereoisomer 2 (chiral HPLC: R1 = 6.52 min): 6.2 mg.
Separation method: HPLC: column: Daicel Chiralpak IE 5 um, 250 mm x 20 mm;
eluent: 50%
n-heptane / 50% 2-propanol; temperature: 40 C; flow rate: 15 ml/min; UV
detection: 235 nm.
Analysis method: HPLC: column: Daicel Chiraltek IE 5 um, 250 mm x 4.6 mm;
eluent: 50% n-
heptane /50% 2-propanol; flow rate: 1.0 ml/min; UV detection: 220 nm.
LC-MS (method 1): R1= 1.08 min; MS (ESIpos): m/z = 618 [M-411+
1H-NMR (500 MHz, DM50-c/6): 6 [ppm] = 10.70 / 10.62 (2br s, 1H), 7.98 (br s,
1H), 7.84 (br d, 3H),
7.70 (br d, 2H), 7.52 (br s, 1H), 7.48-7.32 (m, 2H), 7.24 (br s, 1H), 6.44 /
6.40 (2s, 1H), 5.84-5.72
(m, 1H), 4.82-4.64 (m, 1H), 3.82-3.72 (m, 1H), 3.23-3.17 (m, 1H), 3.13-3.02
(m, 2H), 2.61-2.24 (m,
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1H), 2.04-1.92 (m, 1H), 1.90-1.80 (m, 1H), 1.77-1.29 (m, 6H), 1.28-1.21 (m,
1H), 1.18 (s, 3H).
Additional signals of minor rotamers were also detected.
Example 88
4-(1(2S,4R)-2-R7R)- 11-Chloro-2-oxo-7-(trifluoromethyl)-2,6,7,8 -tetrahydro-3H-
[3] benzoxocino [2,1-c] pyridin-3 -yl] -5,5,5 -trifluoro-4-
hydroxypentanoyllamino)benzamide (single
stereoisomer)
F
FF
0 0
N
H N
0
N H
CI 0
To a solution of
4-[(4-{ [tert-Butyl(dimethypsilylloxy} -2-[(7R)-11-chloro-2-oxo-7-
(trifluoromethyl)-2,6,7,8-tetrahydro-3H-[3]benzoxocino [2,1-c] pyridin-3 -yl] -
5 ,5 ,5 -
trifluoropentanoyl)aminolbenzamide (mixture of two diastereomers) (87.0 mg,
57% purity, 67.7
lama 1.0 eq.) in tetrahydrofuran (4.0 ml) was added a solution of tetra-n-
butylammoniumfluoride
(200
1.0 M in tetrahydrofuran, 200 lama 3.0 eq.) at RT. The mixture was stirred for
3 h and then
concentrated under reduced pressure and further purified by diastereomer
separation to provide:
single stereoisomer 1 (LC-MS (method 5): Rt = 1.27 min): 2.4 mg
single stereoisomer 2 (the title compound 88) (LC-MS (method 5): Rt = 1.28
min, 99% de): 2.1 mg
(5% of theory).
Separation method: HPLC: column: Phenomenex Kinetex C18 5 p.m, 100 x 30 mm;
eluent: 80%
water with 2.0% formic acid /20% acetonitrile; temperature: RT; flow rate: 80
ml/min; UV detection:
220 nm.
.. Analysis method: LC-MS: method 5
LC-MS (method 5): R1= 1.28 min; MS (ESIneg): m/z = 616 EM-H]-
1H-NMR (400 MHz, DM50-c/6): 6 [ppm] = 10.63 (br s, 1H), 7.93 (s, 1H), 7.89-
7.81 (m, 3H), 7.68
(d, 2H), 7.59-7.46 (m, 1H), 7.46-7.34 (m, 2H), 7.26 (br s, 1H), 6.52-6.34 (m,
2H), 5.86-5.66 (m, 1H),
4.75-4.62 (m, 1H), 4.18-4.04 (m, 1H), 4.03-3.87 (m, 1H), 3.18-2.96 (m, 2H),
2.71-2.49 (m, 2H),
.. 2.25-2.09 (m, 1H). Additional signals of minor rotamers were also detected.
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Example 89
4-( { (25)-2 - 1(7 R) - 11 -Chloro-12-fluoro-2 -oxo -7-(trifluoromethyl)-
2,6,7,8-tetrahydro-3H-
[3] benzoxocino [2,1-c] pyridin-3 -yll butanoyllamino)-2-fluorobenzamide
(single stereoisomer)
C H 3
0
N
00 el 0
F N H 2
Ci
General Method 7 was carried out with (7R)-11-chloro-12-fluoro-7-
(trifluoromethyl)-7,8-dihydro-
3H431benzoxocino[2,1-clpyridin-2(6H)-one (single stereoisomer) (60.0 mg, 76%
purity, 131 umol,
1.0 eq.), 4-{[(2R)-2-bromobutanoyllamino}-2-fluorobenzamide (single
stereoisomer) (47.7 mg,
157 umol, 1.2 eq.) and 1,1,3,3-tetramethylguanidine (49 IA, 390 umol, 3.0 eq.)
in a mixture of
2-propanol / acetone (4:1, 1.2 ml) including the following variations of the
procedure: The crude
mixture was purified by preparative HPLC (reversed phase, eluent: water with
0.05% formic acid /
acetonitrile 90:10 to 5:95). Yield: 34.0 mg (41% of theory).
LC-MS (method 1): R1= 1.69 min; MS (ESIpos): m/z = 570 [M+H1+
1H-NMR (400 MHz, DM50-d6): 6 [ppm] = 10.90 / 10.83 (2s, 1H), 8.06 / 8.02 (2s,
1H), 7.74-7.59
(m, 3H), 7.58-7.47 (m, 2H), 7.44-7.36 (m, 1H), 7.34-7.28 (m, 1H), 6.55 / 6.49
(2d, 1H), 5.60-5.50
(m, 1H), 4.79-4.67 (m, 1H), 3.86 / 3.78 (2t, 1H), 3.19-2.97 (m, 2H), 2.60-2.46
(m, 1H), 2.25-2.04
(m, 2H), 0.94-0.91 (2t, 3H). Additional signals of minor rotamers were also
detected.
Example 90
4-( { (25)-2 - 1(7 R) - 11 -Chloro-12-fluoro-2 -oxo -7-(trifluoromethyl)-
2,6,7,8-tetrahydro-3H-
[3] benzoxocino [2,1-c] pyridin-3 -yll butanoyllamino)benzamide (single
stereoisomer)
C H 3
0
N
00 el 0
= F N H 2
C
i
General Method 7 was carried out with (7 R) - 11-chloro-12-fluoro-7-
(trifluoromethyl)-7,8-dihydro-
3H431benzoxocino[2,1-clpyridin-2(6H)-one (single stereoisomer) (60.0 mg, 76%
purity, 131 umol,
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1.0 eq.), 4-{[(2R)-2-bromobutanoyllamino}benzamide (single stereoisomer) (44.9
mg, 157 umol,
1.2 eq.) and 1,1,3,3-tetramethylguanidine (49 jil, 390 umol, 3.0 eq.) in a
mixture of 2-propanol /
acetone (4:1, 1.2 ml) including the following variations of the procedure: The
crude mixture was
purified by preparative HPLC (reversed phase, eluent: water with 0.05% formic
acid / acetonitrile
90:10 to 5:95). Yield: 42.0 mg (57% of theory).
LC-MS (method 3): Rt = 3.22 min; MS (ESIpos): m/z = 552 [M+H1+
1H-NMR (400 MHz, DM50-d6): 6 [ppm] = 10.73 / 10.67 (2s, 1H), 8.07 / 8.03 (2s,
1H), 7.93-7.79
(m, 3H), 7.74-7.60 (m, 3H), 7.31 (d, 1H), 7.27-7.18 (m, 1H), 6.54 /6.48 (2d,
1H), 5.64-5.55 (m, 1H),
4.79-4.67 (m, 1H), 3.78 (t, 1H), 3.30-3.24 (m, 1H), 3.16-2.96 (m, 2H), 2.24-
2.05 (m, 2H), 0.97-0.86
(m, 3H). Additional signals of minor rotamers were also detected.
Example 91
4-{ [(4S)-2 -[(7R)-11 -Chloro-12-fluoro-2 -oxo -7-(trifluoromethyl)-2,6,7,8-
tetrahydro-3H-
[3] benzoxocino [2,1 -c] pyridin-3 -yl] -4-(difluoromethoxy)pentanoyl]
aminolbenzamide (mixture of
two diastereomers)
)<H
0 F
CH3
0
N"(
N-1
00 NH 2
11)1 F 0
CI
(2 )-2- [(7R)-11 -Chloro -12 -fluoro-2-oxo-7-(trifluoromethyl)-2,6,7,8-
tetrahydro-3H-
[3] benzoxocino [2,1 -c] pyridin-3 -yl] -2,3 ,5 -trideoxy-4 -0-
(difluoromethyl)-L -glycero-pentonic acid
(mixture of two diastereomers) (116 mg, 60% purity, 135 umol, 1.0 eq.) was
dissolved in
dichloromethane (5.0 ml), 1-chloro-N,N,2-trimethylprop-1-en-1-amine (23
180 umol, 1.3 eq.)
was added and the reaction mixture was stirred for 10 min. Then 4-
aminobenzamide (24.0 mg, 176
umol, 1.3 eq.) was added and the reaction mixture was stirred for 3 h. All
volatiles were removed
under reduced pressure and the residue was dissolved in NN-dimethylformamide
(4.0 m1). Then
HATU (77.3 mg, 203 umol, 1.5 eq.) and NN-diisopropylethylamine (71
410 umol, 3.0 eq.) were
added and the mixture was stirred at RT overnight and at 50 C for 1 h. The
crude mixture was purified
by preparative HPLC (reversed phase, eluent: water with 0.05% formic acid /
acetonitrile 90:10 to
5:95). Yield: 43.0 mg (49% of theory).
LC-MS (method 3): Rt = 3.40 min; MS (ESIpos): m/z = 632 [M+H1+
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Example 92
4-{[(2S,45)-2-[(7R)- 11-Chloro-12-fluoro-2-oxo-7-(trifluoromethyl)-2,6,7,8-
tetrahydro-3H-
[3] benzoxocino [2,1-c] pyridin-3 -yl] -4-(difluoromethoxy)pentanoyl] amino }
benzamide (single
stereoisomer)
)<H
0 F
C H 3
0
E N
N
00J N H 2
= F 0
CI
Diastereomer separation of 37 mg of 4- { [(4S)-2-[(7R)-11-chloro-12-fluoro-2-
oxo-7-
(trifluoromethyl)-2,6,7,8-tetrahydro-3H-[3]benzoxocino [2,1-c] pyridin-3 -yll -
4 -
(difluoromethoxy)pentanoyllamino}-benzamide (mixture of two diastereomers),
Example 91 gave
single stereoisomer 1 (the title compound 92) (chiral HPLC: Rt = 7.95 min, 97%
de): 9.0 mg (10%
of theory),
single stereoisomer 2 (chiral HPLC: Rt = 10.31 min): 22.0 mg.
Separation method: HPLC: column: Daicel Chiralpak ID 5 p.m, 250 mm x 20 mm;
eluent: 80%
n-heptane / 20% ethanol; temperature: RT; flow rate: 15 ml/min; UV detection:
220 nm.
Analysis method: HPLC: column: Daicel Chiralpak ID 5 p.m, 250 mm x 4.6 mm;
eluent: 80%
iso-hexane /20% ethanol; temperature: 50 C, flow rate: 1.0 ml/min; UV
detection: 220 nm.
LC-MS (method 1): R1= 1.04 min; MS (ESIpos): m/z = 632 [M+I-11+
1H-NMR (500 MHz, DM50-d6): 6 [ppm] = 10.73 / 10.56 (2s, 1H), 8.08 / 8.04 (2s,
1H), 7.92-7.79
(m, 3H), 7.74-7.60 (m, 3H), 7.35-7.27 (m, 1H), 7.27-7.18 (m, 1H), 6.80-6.77 /
6.66-6.60 (2m, 1H),
6.56-6.47 (m, 1H), 5.85 / 5.77 (dd, 1H), 4.78-4.67 (m, 1H), 4.30-4.13 (m, 1H),
3.85-3.71 (m, 1H),
3.14-2.90 (m, 2H), 2.68-2.53 (m, 1H), 2.48-2.33 (m, 2H), 1.38-1.09 (m, 3H).
Additional signals of
minor rotamers were also detected.
Example 93
4-( {(4R)-2 - R7 R) - 11-Chloro-12-fluoro-2-oxo-7-(trifluoromethyl)-2,6,7,8-
tetrahydro-3H-[3]benz-
oxocino [2,1-c] pyridin-3 -yl] -4 -methoxypentanoyl } amino)benzamide (mixture
of two diastereomers)
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H3
0
N
00 N H2
= F 0
CI
(2 )-2- [(7R)-11-Chloro -12 -fluoro-2-oxo-7-(trifluoromethyl)-2,6,7,8-
tetrahydro-3H-
[3] benzoxocino [2,1-c] pyridin-3 -yl] -2,3,5 -trideoxy-4 -0-methyl-D-glyce ro-
pentonic acid (mixture of
two diastereomers) (90.0 mg, 188 umol, 1.0 eq.) was dissolved in
dichloromethane (5.0 ml), 1-
chloro-NN,2-trimethylprop-1-en-1-amine (32 IA, 240 umol, 1.3 eq.) was added
and the reaction
mixture was stirred for 10 min. Then 4-aminobenzamide (33.3 mg, 245 umol, 1.3
eq.) was added
and the reaction mixture was stirred overnight. All volatiles were removed
under reduced pressure
and General Method 11 was carried out with the residue using T3P (170 IA, 50%
solution in ethyl
acetate, 280 umol, 1.5 eq.), pyridine (18 IA, 226 umol, 1.2 eq.) in
tetrahydrofuran (4.0 m1). The crude
mixture was purified by preparative HPLC (reversed phase, eluent: water with
0.05% formic acid /
acetonitrile 90:10 to 5:95). Yield: 58.0 mg (49% of theory).
LC-MS (method 3): Rt = 3.24 min; MS (ESIpos): m/z = 596 [M-411+
Example 94
4-( (2S,4R)-2-R7R)- 11-Chloro-12-fluoro-2 -oxo -7-(trifluoromethyl)-2,6,7,8-
tetrahydro-3H-
[3] benzoxocino [2,1-c] pyridin-3 -yl] -4-methoxypentanoyllamino)benzamide
(single stereoi some r)
Co'CH3
H3
0
N
00 N H2
= F 0
CI
Diastereomer separation of 52 mg of 4-( (4R)-2- [(7R)-11-chloro-12-fluoro-2-
oxo-7-
(trifluoromethyl)-2,6,7, 8-tetrahydro-3H-[3]benz-oxocino [2,1-c] pyridin-3 -
yl] -4-
methoxypentanoyl}amino)benzamide (mixture of two diastereomers), Example 93
gave
single stereoisomer 1 (the title compound 94) (chiral HPLC: Rt = 3.39 min,
>99% de): 13.1 mg (12%
of theory),
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single stereoisomer 2 (chiral HPLC: Rt = 6.91 min): 15.9 mg.
Separation method: HPLC: column: Daicel OZ 5 um, 250 mm x 20 mm; eluent: 50% n-
heptane /
50% 2-propanol; temperature: RT; flow rate: 20 ml/min; UV detection: 220 nm.
Analysis method: HPLC: column: Daicel Chiralpak OX-3 3 pm, 50 mm x 4.6 mm;
eluent: 80%
n-heptane /20% ethanol; flow rate: 1.0 ml/min; UV detection: 220 nm.
LC-MS (method 5): R1= 1.30 min; MS (ESIneg): m/z = 594 EM-H]-
1H-NMR (400 MHz, DM50-d6): 6 [ppm] = 10.72 / 10.62 (2s, 1H), 8.08 / 8.07 (2s,
1H), 7.90-7.81
(m, 3H), 7.73-7.60 (m, 3H), 7.31 (d, 1H), 7.27-7.20 (m, 1H), 6.53 / 6.47 (2d,
1H), 5.86 / 5.80 (2dd,
1H), 4.80-4.67 (m, 1H), 3.77 (t, 1H), 3.20-3.04 (m, 6H), 2.62-2.55 (m, 1H),
2.38-2.19 (m, 2H), 1.20-
1.08 (m, 3H). Additional signals of minor rotamers were also detected.
Example 95
4-(1(2S,45)-2-R7R)- 11-Chl oro-12-fluoro-2 -oxo -7-(trifluoromethyl)-2,6,7,8-
tetrahydro-3H-
[3] benzoxocino [2,1-c] pyridin-3 -yl] -4-methoxypentanoyllamino)benzamide
(single stereoisomer)
C H3
C H3
0
N
00 el N H 2
F 0
C I
General Method 11 was carried out with (2)-2-[(7R)-11-chloro-12-fluoro-2-oxo-7-
(trifluoromethyl)-2,6,7,8-tetrahydro-3H-[3]benzoxocino [2,1-c] pyridin-3 -yll -
2,3 ,5 -trideoxy-4 -0-
methyl-L-glycero-pentonic acid (mixture of two diastereomers) (139 mg, 290
umol, 1.0 eq.), 4-
aminobenzamide (51.3 mg, 377 umol, 1.3 eq.), T3P (259 IA, 50% solution in
ethyl acetate, 435 umol,
1.5 eq.) and pyridine (28 IA, 350 umol, 1.2 eq.) in tetrahydrofuran (8.0 ml)
at RT including the
following variations of the procedure: After 3 h, additional amounts of 4-
aminobenzamide (11.8 mg,
87 umol, 0.3 eq.), pyridine (8.2 IA, 102 umol, 0.35 eq.) and T3P (48 IA, 50%
solution in ethyl acetate,
81 umol, 0.28 eq.) were added and stirring was continued overnight. The crude
mixture was purified
by preparative HPLC (reversed phase, eluent: water with 0.05% formic acid /
acetonitrile 90:10 to
5:95).
Diastereomer separation of 13 mg of 4-( { (45)-2- [(7R)-11 -chloro-12-fluoro-2-
oxo-7-
(trifluoromethyl)-2,6,7, 8-tetrahydro-3H-[3]benzoxocino [2,1-c] pyridin-3 -yll
-4 -
methoxypentanoyl}amino)benzamide (mixture of two diastereomers) gave
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single stereoisomer 1 (the title compound 95) (chiral HPLC: Rt = 5.81 min, 99%
de): 5.2 mg (3% of
theory),
single stereoisomer 2 (chiral HPLC: Rt = 8.47 min): 1.6 mg.
Separation method: HPLC: column: Daicel Chiralpak ID 5 um, 250 mm x 20 mm;
eluent: 80%
n-heptane / 20% ethanol; temperature: 40 C; flow rate: 25 ml/min; UV
detection: 265 nm.
Analysis method: HPLC: column: Daicel Chiraltek ID 3 um, 250 mm x 4.6 mm;
eluent: 80%
n-heptane /20% ethanol; flow rate: 1.0 ml/min; UV detection: 220 nm.
LC-MS (method 3): R1= 1.29 min; MS (ESIneg): m/z = 594 EM-H]-
1H-NMR (500 MHz, CDC13): 6 [ppm] = 9.75 / 9.63 (2br s, 1H), 7.80 (br d, 2H),
7.74-7.67 (m, 2H),
7.60 / 7.59 (2s, 1H), 7.48-7.41 (m, 1H), 7.08 (d, 1H), 6.66 (d, 1H), 5.93 (br
dd, 1H), 6.20-5.58 (m,
2H), 4.75 (dd, 1H), 3.78 (t, 1H), 3.53-3.43 (m, 1H), 3.37-3.27 (m, 3H), 3.06
(br d, 1H), 2.79-2.68
(m, 1H), 2.65-2.53 (m, 2H), 1.97-1.89 (m, 1H), 1.27 (d, 3H). Additional
signals of minor rotamers
were also detected.
Alternatively, General Method 7 was carried out with (7R)-11-chloro-12-fluoro-
7-(trifluoromethyl)-
7,8-dihydro-3H-[3]benzoxocino[2,1-clpyridin-2(6H)-one (single stereoisomer)
(43.0 mg, 124 umol,
1.0 eq.), 4- { [(2R,45)-2 -b romo -4 -methoxypentanoyl] aminolbenzamide
(single stereoisomer)
(44.9 mg, 124 umol, 1.0 eq.) and 1,1,3,3-tetramethylguanidine (47 jil, 371
umol, 3.0 eq.) in a mixture
of 2-propanol / acetone (4:1, 2.0 ml) including the following variations of
the procedure: The crude
mixture was purified by preparative HPLC (reversed phase, eluent: water with
0.05% formic acid /
acetonitrile 80:20 to 5:95). Yield: 42.0 mg (57% of theory).
LC-MS (method 1): Rt = 0.98 min; MS (ESIpos): m/z = 596 [M+I-11+
1H-NMR (500 MHz, CDC13): 6 [ppm] = 9.77 (br s, 1H), 7.80 (br d, 2H), 7.74-7.67
(m, 2H), 7.61 (s,
1H), 7.48-7.41 (m, 1H), 7.08 (d, 1H), 6.66 (d, 1H), 5.93 (br dd, 1H), 6.20-
5.63 (m, 2H), 4.75 (dd,
1H), 3.78 (t, 1H), 3.53-3.44 (m, 1H), 3.37-3.27 (m, 3H), 3.06 (br d, 1H), 2.80-
2.68 (m, 1H), 2.65-
2.53 (m, 2H), 1.97-1.89 (m, 1H), 1.27 (d, 3H). Additional signals of minor
rotamers were also
detected.
Example 96
4-(1(2S,4R)-2-R7R)- 11-Chloro-2-oxo-7-(trifluoromethyl)-2,6,7,8 -tetrahydro-3H-
[3] benzoxocino [2,1-c] pyridin-3 -yl] -5 ,5 ,5 -trifluoro-4-
methoxypentanoyllamino)benzamide (single
stereoisomer)
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F
F.,, F
H 3C'0
0
fee'N)r N
00J N H 2
0
"CI
General Method 7 was carried out with (7R)-11-chloro-7-(trifluoromethyl)-7,8-
dihydro-3H-
[3]benzoxocino[2,1-clpyridin-2(6H)-one (single stereoisomer) (15.0 mg, 45.5
umol, 1.0 eq.),
4-{ [(2R,4R)-2-bromo -5 ,5 ,5 -trifluoro -4 -methoxypentanoyl] aminolbenzamide
(single stereoisomer)
(20.9 mg, 54.6 umol, 1.2 eq.) and 1,1,3,3-tetramethylguanidine (17 jil, 140
umol, 3.0 eq.) in a
mixture of 2-propanol / acetone (4:1, 0.8 ml) including the following
variations of the procedure:
The crude mixture was purified by preparative HPLC (reversed phase, eluent:
water with 0.05%
formic acid / acetonitrile 90:10 to 5:95). Due to epimerization, a mixture of
stereoisomers was
obtained (d.r. 3.6:1). Yield: 16.0 mg (55% of theory).
.. LC-MS (method 3): Rt = 3.62 / 3.67 min; MS (ESIpos): m/z = 632 [M+Hl+
Stereoisomer separation of 16.0 mg of this epimerized mixture provides:
single stereoisomer 1 (chiral HPLC: R1 = 1.25 min): 2.5 mg,
single stereoisomer 2 (the title compound 96) (chiral HPLC: R1 = 2.59 min, 99%
de): 6.5 mg (23%
of theory).
.. Separation method: HPLC: column: Daicel Chiralpak IE 5 um, 250 mm x 20 mm;
eluent: 50%
n-heptane / 50% ethanol; temperature: 40 C; flow rate: 20 ml/min; UV
detection: 210 nm.
Analysis method: HPLC: column: Daicel Chiralpak 1E-3 3 um, 50 mm x 4.6 mm;
eluent: 50%
n-heptane / 50% ethanol; flow rate: 1.0 ml/min; UV detection: 220 nm.
LC-MS (method 3): R1= 3.51; MS (ESIpos): m/z = 632 [M+Hl+
.. 1H-NMR (500 MHz, DM50-d6): 6 [ppm] = 10.69 / 10.48 (2br s, 1H), 8.00 (br s,
1H), 7.93-7.81 (m,
3H), 7.69 (br d, 2H), 7.58-7.44 (m, 1H), 7.44-7.33 (m, 2H), 7.26 (br s, 1H),
6.45 / 6.40 (2br s, 1H),
5.90 / 5.76 (2br s, 1H), 4.79-4.65 (m, 1H), 4.13-3.92 (m, 1H), 3.86-3.71 (m,
1H), 3.40 (s, 3H), 3.06
(br d, 2H), 2.69-2.54 (m, 2H), 2.38-2.16 (m, 1H). Additional signals of minor
rotamers were also
detected.
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Example 97
4-(1(2S,4R)-24(7R)- 11-Chloro-7-methyl-2 -oxo -2,6,7, 8-tetrahydro-3H- [3]
benzoxocino [2,1-
c] pyridin-3 -yll -5 ,5 -difluoro-4 -methoxypentanoyllamino)benzamide (single
stereoisomer)
F.,.. F
H 3C,0
HC 0
NlerN
00J N H
0
CI
1,1,3,3-Tetramethylguanidine (19 0.15 mmol, 3.0 eq.) was added under argon
atmosphere at RT
to a solution of (7 R) - 11-chloro-7-methyl-7,8-dihydro-3H-[3]benzoxocino[2,1-
c]pyridin-2(6H)-one
(single stereoisomer) (14 mg, 0.05 mmol) in 2-propanol / acetone (4:1, 0.5
m1). The mixture was
stirred at RT for 15 min, followed by addition of 4-{[(2R,4R)-2-bromo-5,5-
difluoro-4-
methoxypentanoyllamino}benzamide (single stereoisomer) (21 mg, 0.06 mmol, 1.1
eq.) and further
2-propanol / acetone (4:1, 0.5 m1). The reaction mixture was stirred at RT for
7 days and concentrated
under reduced pressure. The residue was purified by preparative HPLC (reversed
phase, eluent:
acetonitrile / water gradient). Yield: 22 mg (79% of theory).
LC-MS (method 4): R1= 1.86 min; MS (ESIpos): m/z = 560 [M+H1+
'H-NMR (600 MHz, DM50-c/6): 6 [ppm] = 10.84-10.46 (br s, 1H), 7.90-7.77 (m,
4H), 7.73-7.66 (m,
2H), 7.48-7.38 (m, 2H), 7.37-7.28 (m, 1H), 7.23 (br s, 1H), 6.43-6.34 (m, 1H),
6.25-6.00 (m, 1H),
5.91-5.76 (m, 1H), 4.52-4.45 /4.32-4.24 / 3.91-3.81 / 3.59-3.47 (4m, 2H), 3.33
(s, 3H), 2.89-2.82 /
2.75-2.64 (2m, 2H), 2.47-2.15 (m, 2H), 2.14-1.96 (m, 2H), 0.95-0.83 (m, 3H).
Additional signals of
minor rotamers were also detected.
Example 98
4-( {(2S,4R)-2-R7R)- 11-Chloro-2-oxo-7-(trifluoromethyl)-2,6,7,8 -tetrahydro-
3H-
[3] benzoxocino [2,1-c] pyridin-3 -yl] -5 ,5 -difluoro-4 -
methoxypentanoyllamino)benzamide (single
stereoisomer)
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H
F.,. F
H 3C,0
0
1.9.N=rN
00 N H 2
0
Ci
1,1,3,3-Tetramethylguanidine (30 0.24 mmol, 3.0 eq.) was added under argon
atmosphere at RT
to a solution of (7 R) - 11-chloro-7-(trifluoromethyl)-7,8-dihydro-3H-
[3]benzoxocinop,1-clpyridin-
2(611)-one (single stereoisomer) (26 mg, 0.08 mmol) in 2-propanol / acetone
(4:1, 1.0 m1). The
mixture was stirred at RT for 15 min, followed by addition of 4-{[(2R,4R)-2-
bromo-5,5-difluoro-4-
methoxypentanoyllamino}benzamide (single stereoisomer) (33 mg, 0.09 mmol, 1.1
eq.) and further
2-propanol / acetone (4:1, 1.0 m1). The reaction mixture was stirred at RT for
2 days and concentrated
under reduced pressure. The residue was purified by preparative HPLC (reversed
phase, eluent:
acetonitrile / water gradient). Yield: 36 mg (73% of theory).
LC-MS (method 1): R1= 1.00 min; MS (ESIpos): m/z = 614 [M+H1+
'H-NMR (500 MHz, DM50-d6): 6 [ppm] = 10.89-10.38 (m, 1H), 8.07-7.92 (m, 1H),
7.91-7.80 (m,
3H), 7.73-7.65 (m, 2H), 7.56-7.32 (m, 3H), 7.25 (br s, 1H), 6.50-6.36 (m, 1H),
6.26-6.00 (m, 1H),
5.92-5.71 (m, 1H), 4.80-4.64 / 4.11-3.97 / 3.92-3.84 / 3.82-3.72 / 3.64-3.49 /
3.3-3.18 (6m, 3H,
partially concealed), 3.33 (s, 3H), 3.14-2.98 (m, 2H), 2.84-2.74 / 2.65-2.55
(2m, 1H), 2.5-2.40 / 2.38-
2.26 / 2.22-2.08 (3m, 2H, partially concealed). Additional signals of minor
rotamers were also
detected.
Example 99
4-(1(25)-24(7R)- 11 -Chloro-2-oxo -7-(trifluoromethyl)-2,6,7,8-tetrahydro-3H-
[3]benzoxocino [2,1-
c] pyridin-3 -yll propanoyl}amino)-2-fluorobenzamide (single stereoisomer)
FF C H 3 H
0
N.rN
0 F NH2
0
0
CI
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argon atmosphere at RT
to a solution of (7 R) - 11-chloro-7-(trifluoromethyl)-7,8-dihydro-3H-
[3]benzoxocinop,1-clpyridin-
2(611)-one (single stereoisomer) (20 mg, 0.06 mmol) in 2-propanol / acetone
(4:1, 1.0 m1). The
mixture was stirred at RT for 15 min, followed by addition of 4-{[(2R)-2-
bromopropanoyllamino}-
2-fluorobenzamide (single stereoisomer) (19 mg, 0.07 mmol, 1.1 eq.) and of
further 2-propanol /
acetone (4:1, 1.0 m1). The reaction mixture was stirred at RT overnight and
concentrated under
reduced pressure. The residue was purified by preparative HPLC (reversed
phase, eluent: acetonitrile
/ water gradient). Yield: 27 mg (85% of theory).
LC-MS (method 4): R1= 1.82 min; MS (ESIpos): m/z = 538 [M+H1+
'H-NMR (600 MHz, DM50-d6): 6 [ppm] = 10.92-10.36 (br s, 1H), 7.97-7.88 (m,
1H), 7.73-7.66 (m,
1H), 7.66-7.60 (m, 1H), 7.56-7.32 (m, 6H), 6.45-6.32 (m, 1H), 5.57-5.47 (m,
1H), 4.78-4.64 (m, 1H),
4.15-4.05 /3.98-3.77 (2m, 1H), 3.13-2.95 (m, 2H), 2.84-2.73 /2.60-2.5 (2m, 1H,
partially concealed),
1.75-1.64 (m, 3H). Additional signals of minor rotamers were also detected.
Example 100
4-(1(25)-24(7R)- 11 -Chloro-2-oxo -7 -(trifluoromethyl)-2,6,7,8-tetrahydro-3H-
[3]benzoxocino [2,1-
c] pyridin-3 -yll propanoyl}amino)benzamide (single stereoisomer)
C H 3 H
0
0
N
N H 2
0
= 0
Ci
1,1,3,3-Tetramethylguanidine (23 0.18 mmol, 3.0 eq.) was added under argon
atmosphere at RT
to a solution of (7 R) - 11-chloro-7-(trifluoromethyl)-7,8-dihydro-3H-
[3]benzoxocinop,1-clpyridin-
2(61/)-one (single stereoisomer) (20 mg, 0.06 mmol) in 2-propanol / acetone
(4:1, 1.0 m1). The
mixture was stirred at RT for 15 min, followed by addition of 4-{[(2R)-2-
bromopropanoyllamino}benzamide (single stereoisomer) (18 mg, 0.07 mmol, 1.1
eq.) and of further
2-propanol / acetone (4:1, 1.0 m1). The reaction mixture was stirred at RT
overnight and concentrated
under reduced pressure. The residue was purified by preparative HPLC (reversed
phase, eluent:
acetonitrile / water gradient). Yield: 21 mg (68% of theory).
LC-MS (method 4): R1= 1.75 min; MS (ESIpos): m/z = 520 [M+H1+
'H-NMR (400 MHz, DM50-d6): 6 [ppm] = 10.64-10.54 (m, 1H), 7.97-7.90 (m, 1H),
7.90-7.82 (m,
3H), 7.67 (d, 2H), 7.57-7.38 (m, 3H), 7.24 (br s, 1H), 6.44-6.35 (m, 1H), 5.61-
5.52 (m, 1H), 4.80-
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4.64 / 4.16-4.04 / 3.88-3.76 (3m, 2H), 3.14-2.99 (m, 2H), 2.84-2.74 / 2.64-2.5
(2m, 1H, partially
concealed), 1.75-1.64 (m, 3H). Additional signals of minor rotamers were also
detected.
Example 101
(45)-2 -[(7R)-11 -Chloro-2-oxo-7-(trifluoromethyl)-2,6,7, 8-tetrahydro-3H-
[3]benzoxocino [2,1-c] -
pyridin-3-yl] -4 -methoxy-N-(2-methy1-3 -oxo-2,3 -dihydro [1,2,4]triazolo [4,3
-alpyridin-7-y1)-
pentanamide (mixture of two diastereomers)
CYCH3
CH3
0
N
N¨C H 3
0 LN,µ
0
= 0
CI
7-Amino-2-methyl[1,2,41triazolo[4,3-alpyridin-3(2H)-one hydrochloride (25 mg,
0.12 mmol,
1.1 eq.), pyridine (27 jil, 0.34 mmol, 3.0 eq.) and T3P (98 jil, 50% solution
in ethyl acetate,
0.17 mmol, 1.5 eq.) were added under argon atmosphere at RT to a solution of
(2)-2-[(7R)-11-
chloro-2-oxo-7-(trifluoromethyl)-2,6,7,8-tetrahydro-3H-[3]benzoxocino [2,1 -c]
pyridin-3 -yl] -2,3,5 -
trideoxy-4 -0-methyl -L-glycero-pentonic acid (mixture of two diastereomers)
(55 mg, 93% purity,
0.11 mmol) in tetrahydrofuran (4 m1). The reaction mixture was stirred at RT
for 45 min and
concentrated in vacuo. The residue was purified by preparative HPLC (reversed
phase, eluent:
acetonitrile / water gradient). Yield: 59 mg (87% of theory).
LC-MS (method 1): Rt = 0.99 min; MS (ESIpos): m/z = 606 [M+H1+
'H-NMR (600 MHz, DM50-d6): 6 [ppm] = 10.83-10.66 (m, 1H), 8.06-7.96 (m, 1H),
7.86-7.80 (m,
1H), 7.64-7.59 (m, 1H), 7.56-7.32 (m, 3H), 6.75-6.65 (m, 1H), 6.45-6.35 (m,
1H), 5.82-5.67 (m, 1H),
4.78-4.66 (m, 1H), 4.08-3.74 (m, 1H), 3.48 / 3.48 (2s, 3H), 3.3-3.30 (m, 3H,
partially concealed),
3.18 / 3.11 (2s, 3H), 2.84-2.71 /2.60-2.5 (2m, 1H, partially concealed), 2.41-
2.17 (m, 2H), 1.18-1.10
(m, 3H). Additional signals of minor rotamers were also detected.
Example 102
(2S,4S)-2 -[(7R)-11 -Chloro -2-oxo -7 -(trifluoromethyl)-2,6,7,8-tetrahydro-3H-
[3]benzoxocino [2,1-
clpyridin-3-y11-4-methoxy-N-(2-methy1-3-oxo-2,3 -dihydro [1,2,4]triazolo [4,3 -
alpyridin-7 -y1)-
pentanamide (single stereoisomer)
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CYC H3
)_C H3
0
N
N¨C H 3
0
0
= 0
CI
Diastereomer separation of 56 mg of (4S)-2-[(7R)-11-chloro-2-oxo-7-
(trifluoromethyl)-2,6,7,8-
tetrahydro-3H-[3]benzoxocino [2, 1-c] pyridin-3 -yl] -4 -methoxy-N-(2 -methy1-
3 -oxo -2,3 -
dihydro[1,2,41triazolo[4,3-alpyridin-7-yl)pentanamide (mixture of two
diastereomers), Example 101
gave
single stereoisomer 1 (the title compound Example 102) (chiral HPLC: Rt = 8.43
min, >99% de):
16 mg,
single stereoisomer 2 (chiral HPLC: Rt = 10.76 min, >99% de): 30 mg.
Separation method: HPLC: column: Daicel Chiralcel OX-H 5 pm, 250 mm x 20 mm;
eluent: 70%
n-heptane / 30% ethanol; temperature: 40 C; flow rate: 20 ml/min; UV
detection: 220 nm.
Analysis method: HPLC: column: Daicel Chiralcel OX-H 5 pm, 250 mm x 4.6 mm;
eluent: 70%
iso-hexane / 30% ethanol; temperature: 40 C; flow rate: 1 ml/min; UV
detection: 220 nm.
'H-NMR (600 MHz, DM50-d6): 6 [ppm] = 10.89-10.71 (m, 1H), 8.06-7.98 (m, 1H),
7.89-7.83 (m,
1H), 7.68-7.61 (m, 1H), 7.57-7.32 (m, 3H), 6.74-6.67 (m, 1H), 6.46-6.35 (m,
1H), 5.83-5.66 (m, 1H),
4.79-4.66 (m, 1H), 4.08-3.98 / 3.83-3.74 (2m, 1H), 3.48 (s, 3H), 3.30-2.98 (m,
3H), 3.17 (s, 3H),
2.82-2.76 / 2.60-2.5 (2m, 1H, partially concealed), 2.34-2.18 (m, 2H), 1.15
(d, 3H). Additional
signals of minor rotamers were also detected.
Example 103
(45)-2 -[(7R)-11 -Chloro-2-oxo-7-(trifluoromethyl)-2,6,7, 8-tetrahydro-3H-
[3]benzoxocino [2,1-c] -
pyridin-3-yl] -4 -methoxy-N-( [1,2,4]triazolo [4,3 -a] pyridin-7-
yl)pentanamide (mixture of two
diastereomers)
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C H3
C H3
0
N ENlitr-Ns
0
0
=
CI
[1,2,41Triazolo[4,3-alpyridin-7-amine hydrochloride (22 mg, 0.13 mmol, 1.1
eq.), pyridine (28
0.35 mmol, 3.0 eq.) and T3P (102 jil, 50% solution in ethyl acetate, 0.17
mmol, 1.5 eq.) were added
under argon atmosphere at RT to a solution of (2)-2-[(7R)-11-chloro-2-oxo-7-
(trifluoromethyl)-
2,6,7,8-tetrahydro -3H-[3]benzoxocino [2,1 -c] pyridin-3 -yl] -2,3 ,5 -tride
oxy-4-0-methyl-L-glyce ro -
pentonic acid (mixture of two diastereomers) (57 mg, 94% purity, 0.12 mmol) in
tetrahydrofuran
(4 m1). The reaction mixture was stirred at RT overnight. Further
[1,2,41triazolo[4,3-alpyridin-7-
amine hydrochloride (10 mg, 0.06 mmol, 0.5 eq.), pyridine (9 IA, 0.12 mmol,
1.0 eq.) and T3P (102
50% solution in ethyl acetate, 0.17 mmol, 1.5 eq.) were added. The reaction
mixture was stirred
at RT for additional 3 days and concentrated invacuo. The residue was purified
by preparative HPLC
(reversed phase, eluent: acetonitrile / water gradient). Yield: 31 mg (46% of
theory).
LC-MS (method 4): R1= 1.76 min; MS (ESIpos): m/z = 576 [M+H1+
'H-NMR (600 MHz, DM50-c/6): 6 [ppm] = 10.95-10.77 (m, 1H), 9.14/9.14 (2s, 1H),
8.53-8.47 (m,
1H), 8.17-8.10 (m, 1H), 8.08-8.00 (m, 1H), 7.62-7.32 (m, 3H), 7.14-7.05 (m,
1H), 6.46-6.30 (m, 1H),
5.87-5.73 (m, 1H), 4.80-4.60 (m, 1H), 4.10-3.76 (m, 1H), 3.3-2.95 (m, 3H,
partially concealed), 3.19
/3.11 (2s, 3H), 2.84-2.72 / 2.60-2.46 (2m, 1H, partially concealed), 2.43-2.20
(m, 2H), 1.20-1.11 (m,
3H). Additional signals of minor rotamers were also detected.
Example 104
(2S,4S)-2 -[(7R)-11 -Chloro -2-oxo -7 -(trifluoromethyl)-2,6,7,8-tetrahydro-3H-
[3]benzoxocino [2,1-
c] pyridin-3 -y11-4-methoxy-N-([1,2,41triazolo [4,3 -alpyridin-7-
yl)pentanamide (single stereoisomer)
C H3
C H3
0
N ENlirr>--NsN
0
0
CI
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Diastereomer separation of 29 mg of (4S)-2-[(7R)-11-chloro-2-oxo-7-
(trifluoromethyl)-2,6,7,8-
tetrahydro-3H-[3]benzoxocino [2,1-c] pyridin-3 -yl] -4 -methoxy-N-
(j1,2,4]triazolo [4,3 -alpyridin-7-
yl)pentanamide (mixture of two diastereomers), Example 103 gave
single stereoisomer 1 (the title compound Example 104) (chiral HPLC: Rt = 3.11
min, 96% de): 4 mg,
single stereoisomer 2 (chiral HPLC: Rt = 4.15 min, 97% de): 8 mg.
Separation method: HPLC: column: Daicel Chiralcel OZ-H 5 lam, 250 mm x 20 mm;
eluent: 80%
n-heptane / 20% ethanol; temperature: 50 C; flow rate: 30 ml/min; UV
detection: 220 nm.
Analysis method: HPLC: column: Daicel Chiralcel OZ-H 3 lam, 50 mm x 4.6 mm;
eluent: 70%
n-heptane / 30% ethanol; temperature: 40 C; flow rate: 1 ml/min; UV detection:
220 nm.
'H-NMR (400 MHz, DM50-d6): 6 [ppm] = 11.23-11.00 (m, 1H), 9.33 (br s, 1H),
8.69-8.59 (m, 1H),
8.30-8.22 (m, 1H), 8.08-7.97 (m, 1H), 7.57-7.33 (m, 3H), 7.32-7.22 (m, 1H),
6.48-6.33 (m, 1H),
5.86-5.67 (m, 1H), 4.83-4.63 (m, 1H), 4.15-3.76 (m, 1H), 3.3-2.90 (m, 3H,
partially concealed), 3.18
(s, 3H), 2.86-2.74 / 2.61-2.5 (2m, 1H, partially concealed), 2.31-2.21 (m,
2H), 1.17 (m, 3H).
Additional signals of minor rotamers were also detected.
Example 105
(4S)-2-[(7R)-11-Chloro-2-oxo-7-(trifluoromethyl)-2,6,7,8-tetrahydro-3H-
[3]benzoxocino [2,1-ci-
pyridin-3-yl] -4 -methoxy-N-(3 -methyl [1,2,4]triazolo [4,3 -a] pyridin-7-
yl)pentanamide (mixture of
two diastereomers)
Co'CH3
C H 3
0
N
0
0
C H 3
CI
NN-Diisopropylethylamine (54 [11, 0.31 mmol, 3.5 eq.) and a solution of HATU
(51 mg, 0.13 mmol,
1.5 eq.) in NN-dimethylformamide (1 ml) were added under argon atmosphere at
RT to a solution
of (2 )-2- [(7R)-11-chloro -2-oxo -7-(trifluoromethyl)-2,6,7,8-tetrahydro-3H-
[3]benzoxocino [2,1-ci-
pyridin-3-y1]-2,3,5-trideoxy-4-0-methyl-L-glycero-pentonic acid (mixture of
two diastereomers)
(44 mg, 93% purity, 0.09 mmol) and 3-methyl[1,2,41triazolo[4,3-a]pyridin-7-
amine hydrochloride
(21 mg, 0.11 mmol, 1.2 eq.) in NN-dimethylformamide (4 m1). The reaction
mixture was stirred at
RT for 4 days. Further 3-methyl[1,2,41triazolo[4,3-a]pyridin-7-amine
hydrochloride (10 mg, 0.05
mmol, 0.6 eq.), NN-diisopropylethylamine (22 [11, 0.13 mmol, 1.4 eq.) and HATU
(27 mg,
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0.05 mmol, 0.8 eq.) were added. The reaction mixture was stirred at RT
overnight and concentrated
in vacuo. The residue was purified by preparative HPLC (reversed phase,
eluent: acetonitrile / water
gradient). Yield: 11 mg (21% of theory).
'H-NMR (400 MHz, DMSO-d6): 6 [ppm] = 10.92-10.72 (m, 1H), 8.30 (d, 1H), 8.10-
7.98 (m, 2H),
7.57-7.32 (m, 3H), 7.12-7.03 (m, 1H), 6.47-6.35 (m, 1H), 5.82-5.71 (m, 1H),
4.81-4.67 (m, 1H),
4.10-3.75 (m, 1H), 3.3-3.00 (m, 3H), 3.18 / 3.11 (2s, 3H), 2.85-2.73 /2.65-2.5
(2m, 1H, partially
concealed), 2.45-2.18 (m, 2H), 2.42 / 2.41 (2s, 3H), 1.21-1.10 (m, 3H).
Additional signals of minor
rotamers were also detected.
Example 106
(4S)-2-[(7R)-11-Chloro-2-oxo-7-(trifluoromethyl)-2,6,7,8-tetrahydro-3H-
[3]benzoxocino [2,1-ci-
pyridin-3-yl] -4 -methoxy-N-( [1,2,4]triazolo [4,3 -a] pyridin-6 -
yl)pentanamide (mixture of two
diastereomers)
C H3
)_C H3
0
N"'fl NrvrN
0 N'
0
CI
NN-Diisopropylethylamine (30
0.17 mmol, 4.0 eq.) and a solution of HATU (20 mg, 0.05 mmol,
1.2 eq.) in NN-dimethylformamide (1 ml) were added under argon atmosphere at
RT to a solution
of
(2) -2- [(7R)-11 -chloro -2 -oxo -7 -(trifluoromethyl)-2,6,7, 8-tetrahydro-3H-
[3]benzoxocino [2,1-
clpyridin-3-y11-2,3,5-trideoxy-4-0-methyl-L-glycero-pentonic acid (mixture of
two diastereomers)
(20 mg, 0.04 mmol) and [1,2,4]triazolo[4,3-a]pyridin-6-amine hydrochloride (10
mg, 80% purity,
0.05 mmol, 1.1 eq.) in NN-dimethylformamide (1 m1). The reaction mixture was
stirred at RT for
1 h and concentrated in vacuo. The residue was purified by preparative HPLC
(reversed phase,
eluent: acetonitrile / water gradient). Yield: 14 mg (58% of theory).
LC-MS (method 1): Rt = 0.93 min; MS (ESIpos): m/z = 576 [M+H1+
'H-NMR (400 MHz, DM50-d6): 6 [ppm] = 10.84-10.67 (m, 1H), 9.33-9.20 (m, 2H),
8.08-7.95 (m,
1H), 7.79 (d, 1H), 7.57-7.33 (m, 4H), 6.47-6.35 (m, 1H), 5.89-5.75 (m, 1H),
4.81-4.66 (m, 1H), 4.12-
3.71 (m, 1H), 3.3-2.98 (m, 3H, partially concealed), 3.19 / 3.12 (2s, 3H),
2.85-2.72 / 2.64-2.5 (2m,
1H, partially concealed), 2.43-2.17 (m, 2H), 1.21-1.10 (m, 3H). Additional
signals of minor rotamers
were also detected.
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Example 107
4-{2 -[(7R)-11-Chloro -12 -fluoro-2-oxo-7-(trifluoromethyl)-2,6,7,8 -
tetrahydro-3H-[3] benz-
oxocino [2,1 -c] pyridin-3 -yllpropanamido}benzamide (mixture of two
diastereomers)
C H 3 H
0
N.rN
00J N H2
F 0
CI
General Method 7 was carried out with (7R)-11-chloro-12-fluoro-7-
(trifluoromethyl)-7,8-dihydro-
3H-[3]benzoxocino[2,1-clpyridin-2(6H)-one (single stereoisomer) (35.0 mg, 95%
purity, 95.6 [tmol,
1.0 eq.), 4-{[(2R)-2-bromopropanoyllaminolbenzamide (single stereoisomer)
(31.1 mg, 115 lama
1.2 eq.), 1,1,3,3-tetramethylguanidine (36 jil, 290 lama 3.0 eq.) in a mixture
of isopropanol / acetone
(4:1, 1.4 ml) overnight including the following variations of the procedure:
The crude mixture was
purified by preparative HPLC (reversed phase, eluent: water with 0.05% formic
acid / acetonitrile
80:20 to 5:95). Yield: 41.0 mg (80% of theory, mixture of two diastereomers
due to epimerization).
LC-MS (method 3): Rt = 2.78 min; MS (ESIpos): m/z = 538 [M+H1+
1H-NMR (400 MHz, DM50-d6): 6 [ppm] = 10.65 /10.58 (2s, 1H), 8.05 / 8.03 (2s,
1H), 7.90-7.81
(m, 3H), 7.71-7.60 (m, 3H), 7.35-7.28 (m, 1H), 7.24 (br s, 1H), 6.53 / 6.51
(2d, 1H), 5.62-5.52 (m,
1H), 4.77-4.65 (m, 1H), 3.93 / 3.81 (2dd, 1H), 3.17-2.98 (m, 2H), 2.67-2.56
(m, 1H), 1.73 / 1.71 (2d,
3H).
Example 108
4-{ [(2S,45)-2-(11 -Chloro-7-ethyl-2-oxo -2,6,7, 8-tetrahydro -3H-
[3]benzoxocino [2,1 -c] pyridin-3 -y1)-
4-methoxypentanoyllaminolbenzamide (mixture of two diastereomers)
CYCH3
CH3
0
N
H 3C
00J NH2
0
C
i
General Method 7 was carried out with 11-chloro-7-ethy1-7,8-dihydro-3H-
[3]benzoxocinop,1-
clpyridin-2(6H)-one (racemate) (47.0 mg, 66% purity, 107 lama 1.0 eq.), 4-
{[(2R,4S)-2-bromo-4-
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methoxypentanoyllaminolbenzamide (single stereoisomer) (32.5 mg, 98.9 umol,
1.2 eq.), 1,1,3,3-
tetramethylguanidine (40 jil, 320 umol, 3.0 eq.) in a mixture of isopropanol /
acetone (4:1, 1.6 ml)
overnight including the following variations of the procedure: The crude
mixture was purified by
preparative HPLC (reversed phase, eluent: water with 0.05% formic acid /
acetonitrile 90:10 to 5:95).
Yield: 37.0 mg (64% of theory).
LC-MS (method 1): R1= 1.01 min; MS (ESIpos): m/z = 538 [M+I-11+
Example 109
4-(1(2S,45)-24(7R)-11-Chl oro-7-ethy1-2-oxo-2,6,7,8 -tetrahydro-3H- [3]
benzoxocino [2,1-c] pyridin-
3 -yll -4-methoxypentanoyllamino)benzamide (single stereoisomer)
Co'CH3
)_C H3
0
H 3C N
00J N H 2
= 0
C
i
Stereoisomer separation of 37 mg of 4-{ R2S,45)-2-(11-chloro-7-ethyl-2-oxo-
2,6,7,8-tetrahydro-
3H-[3]benzoxocino [2,1 -c] pyridin-3 -y1)-4-methoxypentanoyl] amino benzamide
(mixture of two
diastereomers), Example 108 provided:
single stereoisomer 1 (chiral HPLC: R1 = 2.89 min): 7.5 mg,
single stereoisomer 2 (the title compound Example 109) (chiral HPLC: R1 = 3.90
min): 6.0 mg (10%
of theory, 99% de).
Separation method: column: Daicel Chiralpak IC 3 um, 50 mm x 4.6 mm; eluent:
50% n-heptane /
50% ethanol; temperature: 30 C; flow rate: 10 ml/min; UV detection: 220 nm.
Analysis method: column: Daicel Chiralpak IC 3 um, 50 mm x 4.6 mm; eluent: 50%
n-heptane /
50% ethanol; temperature: 30 C; flow rate: 10 ml/min; UV detection: 220 nm.
LC-MS (method 1): R1= 1.01 min; MS (ESIpos): m/z = 538 [M+I-11+
1H-NMR (500 MHz, DM50-d6): 6 [ppm] = 10.69/ 10.65 (2s, 1H), 7.90-7.80 (m, 4H),
7.70 (d, 2H),
7.46 (d, 1H), 7.40-7.30 (m, 2H), 7.25 (br s, 1H), 6.38 / 6.33 (s, 1H), 5.80
(dd, 1H), 4.53 (dd, 1H),
4.37-4.30 / 3.48-3.35 (2m, 1H), 3.28-3.19 (m, 1H), 3.17 (s, 3H), 2.75 (d, 1H),
2.30-2.12 (m, 3H),
1.84-1.67 (m, 1H), 1.35-1.21 (m, 2H), 1.16 (d, 3H), 1.06 / 0.98 (2t, 3H).
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Example 110
4-( (45)-2 -R7R)-11,12-Dichloro-2-oxo -7-(trifluoromethyl)-2,6,7,8-tetrahydro-
3H-[3]benz-
oxocino[2,1-clpyridin-3-y1]-4-methoxypentanoyl}amino)benzamide (mixture of
stereoisomers)
C H3
H3
0
N
00J N H 2
CI 0
CI
General Method 7 was carried out with (7R)-11,12-dichloro-7-(trifluoromethyl)-
7,8-dihydro-3H-
[3]benzoxocino[2,1-clpyridin-2(6H)-one (single stereoisomer) (20.0 mg, 54.9
umol, 1.0 eq.), 4-
R2R,45)-2-bromo-4-methoxypentanoyllaminolbenzamide (single stereoisomer) (21.7
mg,
65.9 umol, 1.2 eq.), 1,1,3,3-tetramethylguanidine (21 jil, 160 umol, 3.0 eq.)
in a mixture of
isopropanol / acetone (4:1, 0.8 ml) overnight including the following
variations of the procedure:
The crude mixture was purified by preparative HPLC (reversed phase, eluent:
water with 0.05%
formic acid! acetonitrile 90:10 to 5:95). Yield: 37.0 mg (64% of theory).
LC-MS (method 3): Rt = 3.26! 3.33 min; MS (ESIpos): m/z = 612 [M-411+
Example 111
4-(1(2S,45)-2-R7R)-11,12-Dichloro-2-oxo-7-(trifluoromethyl)-2,6,7,8-tetrahydro-
3H-[3]benz-
oxocino[2,1-clpyridin-3-y1]-4-methoxypentanoyl}amino)benzamide (single
stereoisomer)
C H3
)_C H3
0
N
00 N H 2
101 CI 0
CI
Stereoisomer separation of 22.9 mg of 4-({(45)-2-R7R)-11,12-dichloro-2-oxo-7-
(trifluoromethyl)-
2,6,7,8-tetrahydro-3H-[3]benzoxocino[2,1-clpyridin-3-y1]-4-
methoxypentanoyl}amino)benzamide
(mixture of stereoisomers), Example 110 provides:
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single stereoisomer 1 (the title compound Example 111) (chiral HPLC: R1= 1.99
min): 3.7 mg (11%
of theory, 99% de),
single stereoisomer 2 (chiral HPLC: Rt = 2.96 min): 1.8 mg.
Separation method: column: Daicel Chiralpak IB 5 um, 250 mm x 20 mm; eluent:
80% n-heptane /
.. 20% ethanol; temperature: 30 C; flow rate: 20 ml/min; UV detection: 220 nm.
Analysis method: column: Daicel Chiralpak IB 3 um, 50 mm x 4.6 mm; eluent: 80%
n-heptane /
20% ethanol; temperature: 30 C; flow rate: 1.0 ml/min; UV detection: 220 nm.
LC-MS (method 3): Rt = 3.33 min; MS (ESIpos): m/z = 612 [M+H1+
1H-NMR (500 MHz, DM50-d6): 6 [ppm] = 10.62 (s, 1H), 8.11 (s, 1H), 7.89-7.81
(m, 3H), 7.75-7.67
.. (m, 3H), 7.43 (d, 1H), 7.25 (br s, 1H), 6.48 (s, 1H), 5.83 (s, 1H), 4.78-
4.65 (m, 1H), 3.76 (s, 1H),
3.29-3.24 (m, 1H), 3.16 (s, 3H), 3.13-3.00 (m, 2H), 2.63-2.54 (m, 1H), 2.28-
2.19 (m, 2H), 1.15 (d,
3H).
Example 112
4-(1(2S,45)-2-R7R)- 11-Chl oro-7-(difluoromethyl)-2-oxo-2,6,7,8 -tetrahydro-3H-
[3] benzoxocino [2,1-c] pyridin-3 -yl] -4-methoxypentanoyllamino)benzamide
(single stereoisomer)
o-C H 3
CH3
0
N
0 N H 2
0
CI
General Method 7 was carried out with 11-chloro-7-(difluoromethyl)-7,8-dihydro-
3H-
[31benzoxocino[2,1-clpyridin-2(6H)-one (racemate) (550 mg, 1.76 mmol), 4-
{[(2R,45)-2-bromo-4-
methoxypentanoyllaminolbenzamide (single stereoisomer) (987 mg, 3.0 mmol, 1.7
eq.) and 1,1,3,3-
tetramethylguanidine (0.66 ml, 5.29 mmol, 3.0 eq.) in a mixture of 2-propanol
/ acetone (4:1, 20 ml)
for 16 h including the following variations of the procedure: The crude
mixture was treated with
acetic acid (0.3 ml, 5.29 mmol, 3.0 eq.), concentrated under reduced pressure
to 1/3 of the original
volume, treated with 0.02 N aqueous hydrochloric acid (43 ml) and stirred for
0.5 h. The resulting
precipitate was filtered, washed with water and dried.
Diastereomer separation of 1.32 g of the crude product gave:
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single stereoisomer 1 (the title compound Example 112) (chiral SFC: R1 = 4.53
min, >99% de):
309 mg (88% purity, 22% of theory),
single stereoisomer 2 (chiral SFC: R1 = 7.21 min, >99% de): 246 mg (19% of
theory).
Separation method: SFC: column: Daicel Chiralpak IE 5 lam, 250 mm x 20 mm;
eluent: 58% carbon
dioxide / 42% ethanol; temperature: 40 C; flow rate: 85 ml/min; UV detection:
210 nm.
Analysis method: SFC: column: Daicel Chiralpak IE 3 lam, 100 mm x 4.6 mm;
eluent: 70% carbon
dioxide / 30% ethanol; temperature: 40 C; flow rate: 3 ml/min; UV detection:
210 nm.
LC-MS (method 4): R1= 1.76 min; MS (ESIpos): m/z = 560 [M-411+
1H-NMR (500 MHz, DM50-c/6): 6 [ppm] = 10.70 /10.65 (2s, 1H), 7.97 (s, 1H),
7.85 (br d, 3H), 7.70
(br d, 2H), 7.53-7.44 (m, 1H), 7.44-7.31 (m, 2H), 7.25 (br s, 1H), 6.40 / 6.39
(2s, 1H), 6.34-6.06 (m,
1H), 5.86-5.75 (m, 1H), 4.76-4.58 (2m, 1H), 3.72 (t, 1H), 3.28-3.19 (m, 2H),
3.17 (s, 3H), 2.96 (d,
1H), 2.43-2.34 (m, 1H), 2.29-2.15 (m, 2H), 1.15 (br d, 3H). Additional signals
of minor rotamers
were also detected.
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B) Assessment of physio1o2ica1 efficacy
The suitability of the compounds according to the invention for treating
thromboembolic disorders
can be demonstrated in the following assay systems:
a) Test descriptions (in vitro)
a.1) Measurement of FXIa inhibition
The factor XIa inhibition of the substances according to the invention is
determined using a
biochemical test system which utilizes the reaction of a peptidic factor XIa
substrate to determine the
enzymatic activity of human factor XIa. Here, factor XIa cleaves from the
peptidic factor XIa
substrate the C-terminal aminomethylcoumarin (AMC), the fluorescence of which
is measured. The
determinations are carried out in microtitre plates.
Test substances are dissolved in dimethyl sulfoxide and serially diluted in
dimethyl sulfoxide (3000
uM to 0.0078 uM; resulting final concentrations in the test: 50 uM to 0.00013
uM). In each case 1
ul of the diluted substance solutions is placed into the wells of white
microtitre plates from Greiner
(384 wells). 20 pi of assay buffer (50 mM of Tris/HC1 pH 7.4; 100 mM of sodium
chloride; 5 mM
of calcium chloride; 0.1% of bovine serum albumin) and 20 ul of factor XIa
from Kordia (0.45 nM
in assay buffer) are then added successively. After 15 min of incubation, the
enzyme reaction is
started by addition of 20 ul of the factor XIa substrate Boc-Glu(OBz1)-Ala-Arg-
AMC dissolved in
assay buffer (10 uM in assay buffer) from Bachem, the mixture is incubated at
room temperature
(22 C) for 30 min and fluorescence is then measured (excitation: 360 nm,
emission: 460 nm). The
measured emissions of the test batches with test substance are compared to
those of control batches
without test substance (only dimethyl sulfoxide instead of test substance in
dimethyl sulfoxide), and
IC50 values are calculated from the concentration/activity relationships.
Activity data from this test
are listed in Table A below (some as mean values from multiple independent
individual
determinations):
Table A
Example No. I C50 [nIVII Example No. IC51)
1 36 2 25
3 11 4 66
5 3.5 6 2.0
7 2.3 8 0.8
9 4.5 10 1.4
11 2.8 12 3.4
13 6.2 14 14
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Example No. ICs o InMl Example No. IC 50 InIVII
15 50 16 11
17 13 18 3.4
19 3.0 20 1.7
21 81 22 44
23 480 24 81
25 10 26 6.9
27 44 28 15
29 13 30 5.9
31 1.2 32 1.8
33 20 34 7.6
35 55 36 20
37 42 38 32
39 100 40 38
41 14 42 9.8
43 5.8 44 1.8
45 19 46 3.8
47 6.8 48 1.3
49 17 50 5.5
51 10 52 3.5
53 25 54 5.6
55 26 56 8.5
57 45 58 13
59 6.2 60 10
61 4.7 62 5.9
63 0.9 64 100
65 13 66 4.1
67 2.8 68 34
69 25 70 13
71 28 72 6.6
73 31 74 8.1
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Example No. ICso In111l Example No. IC51, InMI
75 4.4 76 2.7
77 5.4 78 2.7
79 6.5 80 3.0
81 11 82 7.4
83 4.9 84 8.9
85 5.4 86 6.5
87 3.1 88 8.3
89 8.9 90 3.0
91 15 92 4.5
93 9.1 94 4.7
95 1.7 96 2.5
97 3.2 98 1.8
99 13 100 4.7
101 42 102 11
103 15 104 14
105 14 106 17
107 6.2 108 7.4
109 5.4 110 21
111 19 112 1.5
a.2) Determination of the selectivity
To demonstrate the selectivity of the substances with respect to FXIa
inhibition, the test substances
are examined for their potential to inhibit other human serine proteases, such
as factor Xa, trypsin
and plasmin. To determine the enzymatic activity of factor Xa (1.3 nmo1/1 from
Kordia), trypsin (83
mU/m1 from Sigma) and plasmin (0.1 ug/m1 from Kordia), these enzymes are
dissolved (50 mmo1/1
of Tris buffer [C,C,C-tris(hydroxymethypaminomethanel, 100 mmo1/1 of NaCl,
0.1% BSA [bovine
serum albumin], 5 mmo1/1 of calcium chloride, pH 7.4) and incubated for 15 min
with test substance
in various concentrations in dimethyl sulfoxide and also with dimethyl
sulfoxide without test
substance. The enzymatic reaction is then started by addition of the
appropriate substrates (5 umo1/1
of Boc-Ile-Glu-Gly-Arg-AMC from Bachem for factor Xa and trypsin, 50 umo1/1 of
Me0Suc-Ala-
Phe-Lys-AMC from Bachem for plasmin). After an incubation time of 30 min at 22
C, fluorescence
is measured (excitation: 360 nm, emission: 460 nm). The measured emissions of
the test mixtures
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with test substance are compared to the control mixtures without test
substance (only dimethyl
sulfoxide instead of test substance in dimethyl sulfoxide) and IC50 values are
calculated from the
concentration/activity relationships.
a.3) Thrombin generation assay (thrombogram)
The effect of the test substances in the thrombin generation assay according
to Hemker is determined
in vitro in human plasma (Octaplas0 from Octapharma).
In the thrombin generation assay according to Hemker, the activity of thrombin
plasma is determined
by measuring the fluorescent cleavage products of the substrate 1-1140 (Z-Gly-
Gly-Arg-AMC,
Bachem). The reactions are carried out in the presence of varying
concentrations of test substance or
the corresponding solvent. To start the reaction, reagents from Thrombinoscope
(30 pM to 0.1 pM
recombinant tissue factor, 24 [IM phospholipids in HEPES) are used. In
addition, a thrombin
calibrator from Thrombinoscope is used, of which the amidolytic activity is
required for calculating
the thrombin activity in a sample containing an unknown amount of thrombin.
The test is carried out
according to the manufacturer's instructions (Thrombinoscope BV): 4 [11 of
test substance or of the
solvent, 76 [11 of plasma and 20 [11 of PPP reagent or thrombin calibrator are
incubated at 37 C for 5
min. After addition of 20 [11 of 2.5 mM thrombin substrate in 20 mM Hepes, 60
mg/ml of BSA, 102
mM of calcium chloride, the thrombin generation is measured every 20 s over a
period of 120 min.
Measurement is carried out using a fluorometer (Fluoroskan Ascent) from Thermo
Electron fitted
with a 390/460 nm filter pair and a dispenser.
Using the Thrombinoscope software, the thrombogram is calculated and
represented graphically. The
following parameters are calculated: lag time, time to peak, peak, ETP
(endogenous thrombin
potential) and start tail.
a.4) Determination of anticoagulatory activity
The anticoagulatory activity of the test substances is determined in vitro in
human plasma and rat
plasma. Fresh whole blood is drawn directly into a mixing ratio of sodium
citrate/blood of 1:9 using
a 0.11 molar sodium citrate solution as receiver. Immediately after the blood
has been drawn, it is
mixed thoroughly and centrifuged at about 4000 g for 15 minutes. The
supernatant is collected as
(platelet-poor) plasma.
The prothrombin time (PT, synonyms: thromboplastin time, quick test) is
determined in the presence
of varying concentrations of test substance or the corresponding solvent using
a commercial test kit
(Neoplastin0 from Boehringer Mannheim or Hemoliance0 RecombiPlastin from
Instrumentation
Laboratory). The test compounds are incubated with plasma at 37 C for 3
minutes. Coagulation is
then started by addition of thromboplastin, and the timepoint, at which
clotting of the sample occurs
is determined. The concentration of test substance which effects a doubling of
the prothrombin time
is determined.
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The activated partial thromboplastin time (APTT) is determined in the presence
of varying
concentrations of test substance or the corresponding solvent using a
commercial test kit (PTT
reagent from Roche). The test compounds are incubated with the plasma and the
PTT reagent
(cephalin, kaolin) at 37 C for 3 minutes. Coagulation is then started by
addition of 25 mM calcium
chloride, and the time when coagulation occurs is determined. The
concentration of test substance
which leads to an extension by 50% or a doubling of the APTT is determined.
a.5) Determination of the plasma kallikrein activity
To determine the plasma kallikrein inhibition of the substances according to
the invention, a
biochemical test system is used which utilizes the reaction of a peptidic
plasma kallikrein substrate
to determine the enzymatic activity of human plasma kallikrein. Here, plasma
kallikrein cleaves from
the peptidic plasma kallikrein substrate the C-terminal aminomethylcoumarin
(AMC), the
fluorescence of which is measured. The determinations are carried out in
microtitre plates.
Test substances are dissolved in dimethyl sulfoxide and serially diluted in
dimethyl sulfoxide (3000
uM to 0.0078 uM; resulting final concentrations in the test: 50 uM to 0.00013
uM). In each case 1
ul of the diluted substance solutions is placed into the wells of white
microtitre plates from Greiner
(384 wells). 20 ul of assay buffer (50 mM Tris/HC1 pH 7.4; 100 mM sodium
chloride solution; 5
mM of calcium chloride solution; 0.1% of bovine serum albumin) and 20 pi of
plasma kallikrein
from Kordia (0.6 nM in assay buffer) are then added successively. After 15 min
of incubation, the
enzyme reaction is started by addition of 20 ul of the substrate H-Pro-Phe-Arg-
AMC dissolved in
assay buffer (10 uM in assay buffer) from Bachem, the mixture is incubated at
room temperature
(22 C) for 30 min and fluorescence is then measured (excitation: 360 nm,
emission: 460 nm). The
measured emissions of the test batches with test substance are compared to
those of control batches
without test substance (only dimethyl sulfoxide instead of test substance in
dimethyl sulfoxide), and
IC50 values are calculated from the concentration/activity relationships.
Activity data from this test
.. are listed in Table B below (some as mean values from multiple independent
individual
determinations):
Table B
Example No. IC50 InM] Example No. IC5Il in1141
1 760 2 440
3 250 4 190
5 200 6 170
7 150 8 51
9 330 10 110
11 120 12 210
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Example No. ICs o InMl Example No. IG,I, InINII
13 93 14 180
15 850 16 210
17 460 18 130
19 110 20 60
21 8700 22 1500
23 >10000 24 3900
25 2000 26 1300
27 540 28 170
29 190 30 91
31 47 32 40
33 660 34 280
35 300 36 380
37 510 38 780
39 >10000 40 950
41 520 42 340
43 340 44 170
45 890 46 220
47 430 48 120
49 330 50 130
51 160 52 59
53 370 54 110
55 250 56 130
57 250 58 89
59 630 60 230
61 160 62 150
63 35 64 850
65 180 66 190
67 130 68 550
69 430 70 310
71 390 72 110
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Example No. ICso InMi Example No. IC51, InMI
73 520 74 190
75 120 76 95
77 210 78 140
79 130 80 65
81 330 82 310
83 190 84 180
85 150 86 250
87 160 88 460
89 380 90 130
91 190 92 71
93 200 94 110
95 44 96 220
97 110 98 110
99 480 100 190
101 430 102 130
103 190 104 170
105 720 106 490
107 200 108 660
109 350 110 1000
111 870 112 98
b) Determination of antithrombotic activity (in vivo)
b.1) Arterial thrombosis model (iron(II) chloride-induced thrombosis) in
combination with ear
bleeding time in rabbits
The antithrombotic activity of the FXIa inhibitors is tested in an arterial
thrombosis model. Thrombus
formation is triggered here by causing chemical injury to a region in the
carotid artery in rabbits.
Simultaneously, the ear bleeding time is determined.
Male rabbits (Crl:KBL (NZW)BR, Charles River) receiving a normal diet and
having a body weight
of 2.2 ¨ 2.5 kg are anaesthetized by intramuscular administration of xylazine
and ketamine (Rompun,
Bayer, 5 mg/kg and Ketavet, Pharmacia & Upjohn GmbH, 40 mg/kg body weight).
Anaesthesia is
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maintained by intravenous administration of the same preparations (continuous
infusion) via the right
auricular vein.
The right carotid artery is exposed and the vessel injury is caused by
wrapping a piece of filter paper
(10 mm x 10 mm) on a Parafilm0 strip (25 mm x 12 mm) around the carotid artery
without disturbing
the blood flow. The filter paper contains 100 iaL of a 13% strength solution
of iron(II) chloride
(Sigma) in water. After 5 min, the filter paper is removed and the vessel is
rinsed twice with aqueous
0.9% strength sodium chloride solution. 30 min after the injury the injured
region of the carotid artery
is extracted surgically and any thrombotic material is removed and weighed.
The test substances are administered either intravenously to the anaesthetized
animals via the femoral
vein or orally to the awake animals via gavage, in each case 5 min and 2 h,
respectively, before the
injury.
Ear bleeding time is determined 2 min after injury to the carotid artery. To
this end, the left ear is
shaved and a defined 3-mm-long incision (blade Art. Number 10-150-10, Martin,
Tuttlingen,
Germany) is made parallel to the longitudinal axis of the ear. Care is taken
not to damage any visible
vessels. Any blood that extravasates is taken up in 15 second intervals using
accurately weighed filter
paper pieces, without touching the wound directly. Bleeding time is calculated
as the time from
making the incision to the point in time when no more blood can be detected on
the filter paper. The
volume of the extravasated blood is calculated after weighing of the filter
paper pieces.
c) Determination of permeability (Caco assay)
The Caco cells (obtained from the Deutsche Sammlung fur Mikroorganismen and
Zellkulturen,
DSMZ) are cultivated in 24-well Transwell plates for 15 or 16 days. The test
is carried out using a
Hamilton robot. The density of the cell monolayers is ensured by measuring the
Lucifer yellow
permeability. The test compounds are dissolved in DMSO and then diluted with
assay buffer to a
concentration of 2 1.1M (final DMSO concentration 1%). The permeability is
examined in both
directions by addition of the substance solutions to the apical or basolateral
compartment. The
covered plates are incubated at 37 C for 2 hours. The concentrations in the
two compartments are
determined by LC-MS/MS and the Papp values are calculated according to
Artursson and Karlsson
(PMID: 1673839).
d) Determination of pharmacokinetic parameters f0110win2 intravenous
administration
To examine the pharmacokinetic properties of a test substance, the respective
test substances are
administered to animals as a bolus injection, infusion or via oral
administration. In the case of rats,
the preferred formulation for intravenous administration of the test
substances is plasma/dimethyl
sulfoxide in a ratio of 99:1. The infusion solution of the test substance in
the case of dogs and
monkeys consists of polyethylene glycol/ethanol/water in a ratio of 50/10/40.
Formulations for oral
administration can be polyethylene glycol/ethanol/water or
solutol/ethanol/water in a ratio of
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50/10/40, or other formulations as appropriate (e.g. water, tylose, self-
emulsifying drug dispering
systems, etc.). The administration volume for rats is 2-10 ml/kg, for dogs and
monkeys 0.5-5 ml/kg.
Blood samples are removed from the test animals into sodium EDTA (or other
anticoagulant)-
containing tubes: in the case of bolus administration, blood samples are
usually taken at 0.033, 0.083,
0.167, 0.25, 0.283, 0.333, 0.5, 0.75, 1, 2, 3, 5, 7, 24 hours after
administration of the test substance.
In the case of infusions, blood samples are usually taken at 0.083, 0.167,
0.25, 0.283, 0.333, 0.5,
0.75, 1, 2, 3, 5, 7, 24 hours after administration of the test substance. In
the case of oral administration,
blood samples are usually taken at 0.083, 0.25, 0.5, 0.75, 1, 2, 3, 5, 7, 24
hours after administration
of the test substance. Other time points might be chosen as appropriate.
After removal, the blood samples are centrifuged at 1280 g for 10 minutes. The
supernatant (plasma)
is taken off and either directly processed further or frozen for later sample
preparation. For sample
preparation, 50 pi of plasma are mixed with 250 pi of acetonitrile (the
precipitating agent acetonitrile
also contains the internal standard ISTD for later analytical determination)
and then allowed to stand
at room temperature for 5 minutes. The mixture is then centrifuged at 16 000 g
for 3 minutes. The
supernatant is taken off, and 500 pi of a buffer suitable for the mobile phase
are added. The samples
are then examined by LC-MS/MS analysis (e.g. liquid chromatography using a
Gemini 5 [tM C18
110A 50 mm x 3 mm (or 150 mm x 3 mm) column from Phenomenex; by mass
spectrometry using
an API 5500 or API 6500; SCIEX, Canada) to determine the concentration of the
test substance in
the individual samples.
In addition to the the plasma concentrations, the concentration ratio whole
blood to plasma for the
test substance in question is determined. To this end, the test substance is
incubated at a certain
concentration in whole blood for 20 minutes. The samples are then processed as
described above to
determine the concentration of the test substance in the plasma. The
concentration set divided by the
concentration measured in the plasma gives the parameter Cb/Cp.
The pharmacokinetic parameters are calculated by non-compartmental analysis
(NCA). The
algorithms for calculating the parameters are defined in an internal process
description and are based
on rules published in general textbooks of pharmacokinetics.
The primary pharmacokinetic parameters clearance (CL) and distribution volume
(Vss) are
calculated as follows:
Parameter Formula
CLplasma (plasma clearance) CLplasma = dose / AUC (AUC = area under the
curve)
CLblood (blood clearance) CLblood = CLplasma / (Cb/Cp)
Vss Vss = CLplasma * MRTiv
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Parameter Formula
MRTiv MRTiv = AUMC/AUC
AUMC AUMC = AUMC(0 t t -. Last, + 4ast*
Clast,calculatedaz Clast,calculatedi
Rate constant for the terminal phase; calculated from the
logarithmic-linear regression of unweighted data from the
terminal phase with data points above the detection limit
AUC AUC = AUC(0-tlast) + Clast,cakulatedaz
AUCnorm AUC divided by dose (mg) per kg body weight
C) Workin2 examples of pharmaceutical compositions
The substances according to the invention can be converted to pharmaceutical
preparations as
follows:
Tablet:
Composition:
100 mg of the compound of Example 1, 50 mg of lactose (monohydrate), 50 mg of
maize starch, 10
mg of polyvinylpyrrolidone (PVP 25) (from BASF, Germany) and 2 mg of magnesium
stearate.
Tablet weight 212 mg. Diameter 8 mm, radius of curvature 12 mm.
Production:
The mixture of the compound of Example 1, lactose and starch is granulated
with a 5% strength
solution (m/m) of the PVP in water. After drying, the granules are mixed with
the magnesium stearate
for 5 min. This mixture is compressed in a conventional tabletting press (see
above for format of the
tablet).
Oral suspension:
Composition:
1000 mg of the compound of Example 1, 1000 mg of ethanol (96%), 400 mg of
Rhodigel (xanthan
gum) (from FMC, USA) and 99 g of water.
10 ml of oral suspension correspond to a single dose of 100 mg of the compound
of the invention.
Production:
The Rhodigel is suspended in ethanol, and the compound of Example 1 is added
to the suspension.
The water is added while stirring. The mixture is stirred for about 6 h until
swelling of the Rhodigel
is complete.
