U.S. patent application number 15/523061 was filed with the patent office on 2017-11-02 for hydroxyalkyl-substituted phenyltriazole derivatives and uses thereof.
The applicant listed for this patent is BAYER PHARMA AKTIENGESELLSCHAFT. Invention is credited to Hartmut BECK, Anna ENGELEN, Michael GERISCH, Nils GRIEBENOW, Peter KOLKHOF, Florian KOLLING, Axel KRETSCHMER, Dieter LANG, Klemens LUSTIG, Thomas MONDRITZKI, Elisabeth POOK, Carsten SCHMECK, Frank SU MEIER, Sonja VOLLMER, Pierre WASNAIRE.
Application Number | 20170313665 15/523061 |
Document ID | / |
Family ID | 51897095 |
Filed Date | 2017-11-02 |
United States Patent
Application |
20170313665 |
Kind Code |
A1 |
SCHMECK; Carsten ; et
al. |
November 2, 2017 |
HYDROXYALKYL-SUBSTITUTED PHENYLTRIAZOLE DERIVATIVES AND USES
THEREOF
Abstract
The present invention relates to novel
5-(hydroxyalkyl)-1-phenyl-1,2,4-triazole derivatives, to processes
for the preparation of such compounds, to pharmaceutical
compositions containing such compounds, and to the use of such
compounds or compositions for the treatment and/or prevention of
diseases, in particular for the treatment and/or prevention of
cardiovascular and renal diseases.
Inventors: |
SCHMECK; Carsten; (Mulheim,
DE) ; GERISCH; Michael; (Wuppertal, DE) ;
GRIEBENOW; Nils; (Dormagen, DE) ; KOLKHOF; Peter;
(Wuppertal, DE) ; KOLLING; Florian; (Wuppertal,
DE) ; ENGELEN; Anna; (Essen, DE) ; KRETSCHMER;
Axel; (Wuppertal, DE) ; LANG; Dieter;
(Velbert, DE) ; LUSTIG; Klemens; (Wuppertal,
DE) ; MONDRITZKI; Thomas; (Velbert, DE) ;
POOK; Elisabeth; (Wuppertal, DE) ; BECK; Hartmut;
(Wuppertal, DE) ; SU MEIER; Frank; (Munchen,
DE) ; VOLLMER; Sonja; (Kleinmachnow, DE) ;
WASNAIRE; Pierre; (Dusseldorf, DE) |
|
Applicant: |
Name |
City |
State |
Country |
Type |
BAYER PHARMA AKTIENGESELLSCHAFT |
Berlin |
|
DE |
|
|
Family ID: |
51897095 |
Appl. No.: |
15/523061 |
Filed: |
October 30, 2015 |
PCT Filed: |
October 30, 2015 |
PCT NO: |
PCT/EP2015/075200 |
371 Date: |
April 28, 2017 |
Current U.S.
Class: |
1/1 |
Current CPC
Class: |
A61K 31/4196 20130101;
A61P 9/00 20180101; C07D 403/06 20130101; A61K 45/06 20130101; A61P
9/04 20180101; A61P 13/12 20180101; A61P 43/00 20180101; C07D
249/12 20130101; A61P 1/16 20180101; A61P 3/12 20180101; A61P 3/00
20180101; A61P 7/10 20180101; A61P 13/00 20180101; A61P 7/00
20180101 |
International
Class: |
C07D 249/12 20060101
C07D249/12; A61K 45/06 20060101 A61K045/06; A61K 31/4196 20060101
A61K031/4196 |
Foreign Application Data
Date |
Code |
Application Number |
Nov 3, 2014 |
EP |
14191491.1 |
Claims
1. A compound of formula (I) ##STR00059## wherein R.sup.1 is
hydrogen or methyl, and R.sup.2A and R.sup.2B are independently
selected from the group consisting of hydrogen, fluoro, chloro,
cyano, methyl, fluoromethyl, difluoromethyl, trifluoromethyl,
ethyl, methoxy, difluoromethoxy, and trifluoromethoxy; or a
pharmaceutically acceptable salt, hydrate, and/or solvate
thereof.
2. The compound of formula (I) according to claim 1, wherein
R.sup.1 is hydrogen or methyl, and R.sup.2A and R.sup.2B are
independently selected from the group consisting of hydrogen,
fluoro, chloro, methyl, and methoxy, wherein at least one of
R.sup.2A and R.sup.2B is other than hydrogen; or a pharmaceutically
acceptable salt, hydrate, and/or solvate thereof.
3. The compound of formula (I) according to claim 1, wherein the
compound is selected from the group consisting of
5-(4-chlorophenyl)-2-{[1-(3-chlorophenyl)-5-(hydroxymethyl)-1H-1,2,4-tria-
zol-3-yl]methyl}-4-[(2S)-3,3,3-trifluoro-2-hydroxypropyl]-2,4-dihydro-3H-1-
,2,4-triazol-3-one;
5-(4-chlorophenyl)-2-{[1-(3-fluorophenyl)-5-(hydroxymethyl)-1H-1,2,4-tria-
zol-3-yl]methyl}-4-[(2S)-3,3,3-trifluoro-2-hydroxypropyl]-2,4-dihydro-3H-1-
,2,4-triazol-3-one;
5-(4-chlorophenyl)-2-{[5-(hydroxymethyl)-1-(2-methylphenyl)-1H-1,2,4-tria-
zol-3-yl]methyl}-4-[(2S)-3,3,3-trifluoro-2-hydroxypropyl]-2,4-dihydro-3H-1-
,2,4-triazol-3-one;
2-({1-(2-chloro-4-fluorophenyl)-5-[(1RS)-1-hydroxyethyl]-1H-1,2,4-triazol-
-3-yl}methyl)-5-(4-chlorophenyl)-4-[(2S)-3,3,3-trifluoro-2-hydroxypropyl]--
2,4-dihydro-3H-1,2,4-triazol-3-one;
2-{[1-(2-chloro-4-fluorophenyl)-5-(1-hydroxyethyl)-1H-1,2,4-triazol-3-yl]-
methyl}-5-(4-chlorophenyl)-4-[(2S)-3,3,3-trifluoro-2-hydroxypropyl]-2,4-di-
hydro-3H-1,2,4-triazol-3-one (diastereomer 1);
2-{[1-(2-chloro-4-fluorophenyl)-5-(1-hydroxyethyl)-1H-1,2,4-triazol-3-yl]-
methyl}-5-(4-chlorophenyl)-4-[(2S)-3,3,3-trifluoro-2-hydroxypropyl]-2,4-di-
hydro-3H-1,2,4-triazol-3-one (diastereomer 2);
2-({1-(2-chloro-5-fluorophenyl)-5-[(1RS)-1-hydroxyethyl]-1H-1,2,4-triazol-
-3-yl}methyl)-5-(4-chlorophenyl)-4-[(2S)-3,3,3-trifluoro-2-hydroxypropyl]--
2,4-dihydro-3H-1,2,4-triazol-3-one;
2-{[1-(2-chloro-5-fluorophenyl)-5-(1-hydroxyethyl)-1H-1,2,4-triazol-3-yl]-
methyl}-5-(4-chlorophenyl)-4-[(2S)-3,3,3-trifluoro-2-hydroxypropyl]-2,4-di-
hydro-3H-1,2,4-triazol-3-one (diastereomer 1);
2-{[1-(2-chloro-5-fluorophenyl)-5-(1-hydroxyethyl)-1H-1,2,4-triazol-3-yl]-
methyl}-5-(4-chlorophenyl)-4-[(2S)-3,3,3-trifluoro-2-hydroxypropyl]-2,4-di-
hydro-3H-1,2,4-triazol-3-one (diastereomer 2);
5-(4-chlorophenyl)-2-({1-(3-fluorophenyl)-5-[(1RS)-1-hydroxyethyl]-1H-1,2-
,4-triazol-3-yl}methyl)-4-[(2S)-3,3,3-trifluoro-2-hydroxypropyl]-2,4-dihyd-
ro-3H-1,2,4-triazol-3-one;
5-(4-chlorophenyl)-2-({1-(3-fluorophenyl)-5-[(1R)-1-hydroxyethyl]-1H-1,2,-
4-triazol-3-yl}methyl)-4-[(2S)-3,3,3-trifluoro-2-hydroxypropyl]-2,4-dihydr-
o-3H-1,2,4-triazol-3-one;
5-(4-chlorophenyl)-2-({1-(3-fluorophenyl)-5-[(1S)-1-hydroxyethyl]-1H-1,2,-
4-triazol-3-yl}methyl)-4-[(2S)-3,3,3-trifluoro-2-hydroxypropyl]-2,4-dihydr-
o-3H-1,2,4-triazol-3-one;
5-(4-chlorophenyl)-2-({1-(3-chlorophenyl)-5-[(1RS)-1-hydroxyethyl]-1H-1,2-
,4-triazol-3-yl}methyl)-4-[(2S)-3,3,3-trifluoro-2-hydroxypropyl]-2,4-dihyd-
ro-3H-1,2,4-triazol-3-one;
5-(4-chlorophenyl)-2-({1-(3-chlorophenyl)-5-[(1R)-1-hydroxyethyl]-1H-1,2,-
4-triazol-3-yl}methyl)-4-[(2S)-3,3,3-trifluoro-2-hydroxypropyl]-2,4-dihydr-
o-3H-1,2,4-triazol-3-one;
5-(4-chlorophenyl)-2-({1-(3-chlorophenyl)-5-[(1S)-1-hydroxyethyl]-1H-1,2,-
4-triazol-3-yl}methyl)-4-[(2S)-3,3,3-trifluoro-2-hydroxypropyl]-2,4-dihydr-
o-3H-1,2,4-triazol-3-one;
5-(4-chlorophenyl)-2-({1-(2-chlorophenyl)-5-[(1RS)-1-hydroxyethyl]-1H-1,2-
,4-triazol-3-yl}methyl)-4-[(2S)-3,3,3-trifluoro-2-hydroxypropyl]-2,4-dihyd-
ro-3H-1,2,4-triazol-3-one;
5-(4-chlorophenyl)-2-({1-(2-chlorophenyl)-5-[(1R)-1-hydroxyethyl]-1H-1,2,-
4-triazol-3-yl}methyl)-4-[(2S)-3,3,3-trifluoro-2-hydroxypropyl]-2,4-dihydr-
o-3H-1,2,4-triazol-3-one; and
5-(4-chlorophenyl)-2-({1-(2-chlorophenyl)-5-[(1S)-1-hydroxyethyl]-1H-1,2,-
4-triazol-3-yl}methyl)-4-[(2S)-3,3,3-trifluoro-2-hydroxypropyl]-2,4-dihydr-
o-3H-1,2,4-triazol-3-one; or a pharmaceutically acceptable salt,
hydrate, and/or solvate thereof.
4. The compound of formula (I) according to claim 1, wherein the
compound is selected from the group consisting of
5-(4-chlorophenyl)-2-({1-(3-fluorophenyl)-5-[(1RS)-1-hydroxyethyl]-1H-1,2-
,4-triazol-3-yl}methyl)-4-[(2S)-3,3,3-trifluoro-2-hydroxypropyl]-2,4-dihyd-
ro-3H-1,2,4-triazol-3-one;
5-(4-chlorophenyl)-2-({1-(3-fluorophenyl)-5-[(1S)-1-hydroxyethyl]-1H-1,2,-
4-triazol-3-yl}methyl)-4-[(2S)-3,3,3-trifluoro-2-hydroxypropyl]-2,4-dihydr-
o-3H-1,2,4-triazol-3-one;
5-(4-chlorophenyl)-2-({1-(3-chlorophenyl)-5-[(1RS)-1-hydroxyethyl]-1H-1,2-
,4-triazol-3-yl}methyl)-4-[(2S)-3,3,3-trifluoro-2-hydroxypropyl]-2,4-dihyd-
ro-3H-1,2,4-triazol-3-one;
5-(4-chlorophenyl)-2-({1-(3-chlorophenyl)-5-[(1S)-1-hydroxyethyl]-1H-1,2,-
4-triazol-3-yl}methyl)-4-[(2S)-3,3,3-trifluoro-2-hydroxypropyl]-2,4-dihydr-
o-3H-1,2,4-triazol-3-one;
5-(4-chlorophenyl)-2-({1-(2-chlorophenyl)-5-[(1RS)-1-hydroxyethyl]-1H-1,2-
,4-triazol-3-yl}methyl)-4-[(2S)-3,3,3-trifluoro-2-hydroxypropyl]-2,4-dihyd-
ro-3H-1,2,4-triazol-3-one; and
5-(4-chlorophenyl)-2-({1-(2-chlorophenyl)-5-[(1S)-1-hydroxyethyl]-1H-1,2,-
4-triazol-3-yl}methyl)-4-[(2S)-3,3,3-trifluoro-2-hydroxypropyl]-2,4-dihydr-
o-3H-1,2,4-triazol-3-one; or a pharmaceutically acceptable salt,
hydrate, and/or solvate thereof.
5. Process for preparing a compound of formula (I) as defined in
claim 1, wherein a compound of formula (II) ##STR00060## is first
reacted with hydrazine to give the hydrazide of formula (III)
##STR00061## then condensed with an amidine of formula (IV)
##STR00062## or a salt thereof, in the presence of a base to give a
1,2,4-triazole derivative of formula (V) ##STR00063## and/or a
tautomer thereof, and subsequently coupled with a phenylboronic
acid of formula (VI) ##STR00064## in the presence of a copper
catalyst and an amine base to yield the target compound of formula
(I) ##STR00065## optionally followed, where appropriate, by (i)
separating the compounds of formula (I) thus obtained into their
respective diastereomers, and/or (ii) converting the compounds of
formula (I) into their respective hydrates, solvates, salts, and/or
hydrates or solvates of the salts by treatment with the
corresponding solvents and/or acids or bases.
6. Compound as defined in claim 1 for the treatment and/or
prevention of diseases.
7. Compound as defined in claim 1 for use in a method for the
treatment and/or prevention of acute and chronic heart failure,
cardiorenal syndrome, hypervolemic and euvolemic hyponatremia,
liver cirrhosis, ascites, edema, or the syndrome of inadequate ADH
secretion (SIADH).
8. Use of a compound as defined in claim 1 for the manufacture of a
pharmaceutical composition for the treatment and/or prevention of
acute and chronic heart failure, cardiorenal syndrome, hypervolemic
and euvolemic hyponatremia, liver cirrhosis, ascites, edema, or the
syndrome of inadequate ADH secretion (SIADH).
9. Pharmaceutical composition comprising a compound as defined in
claim 1, and one or more pharmaceutically acceptable
excipients.
10. The pharmaceutical composition of claim 9 further comprising
one or more additional therapeutic agents selected from the group
consisting of diuretics, angiotensin AII antagonists, ACE
inhibitors, beta-receptor blockers, mineralocorticoid receptor
antagonists, organic nitrates, NO donors, activators of the soluble
guanylate cyclase, stimulators of the soluble guanylate cyclase and
positive-inotropic agents.
11. The pharmaceutical composition as defined in claim 9 for the
treatment and/or prevention of acute and chronic heart failure,
cardiorenal syndrome, hypervolemic and euvolemic hyponatremia,
liver cirrhosis, ascites, edema, or the syndrome of inadequate ADH
secretion (SIADH).
12. Method for the treatment and/or prevention of acute and chronic
heart failure, cardiorenal syndrome, hypervolemic and euvolemic
hyponatremia, liver cirrhosis, ascites, edema, or the syndrome of
inadequate ADH secretion (SIADH) in a human or other mammal;
comprising administering to the human or other mammal in need
thereof a therapeutically effective amount of one or more compounds
as defined in claim 1; or of a pharmaceutical composition
comprising the compound, and one or more pharmaceutically
acceptable excipients.
Description
[0001] The present invention relates to novel
5-(hydroxyalkyl)-1-phenyl-1,2,4-triazole derivatives, to processes
for the preparation of such compounds, to pharmaceutical
compositions containing such compounds, and to the use of such
compounds or compositions for the treatment and/or prevention of
diseases, in particular for the treatment and/or prevention of
cardiovascular and renal diseases.
[0002] The liquid content of the human body is subject to various
physiological control mechanisms, the purpose of which is to keep
it constant (volume homeostasis). In the process, both the volume
filling of the vascular system and also the osmolarity of the
plasma are continuously recorded by appropriate sensors
(baroreceptors and osmoreceptors). The information which these
sensors supply to the relevant centers in the brain regulates
drinking behaviour and controls fluid excretion via the kidneys by
means of humoral and neural signals. The peptide hormone
vasopressin is of central importance in this [Schrier R. W.,
Abraham W. T., New Engl. J. Med. 341, 577-585 (1999)].
[0003] Vasopressin is produced in specialized endocrine neurons in
the Nucleus supraopticus and N. paraventricularis in the wall of
the third ventricle (hypothalamus) and is transported from there
along the neural processes into the posterior lobes of the
hypophysis (neurohypophysis). There the hormone is released into
the bloodstream in response to stimulus. A loss of volume, e.g. as
a result of acute bleeding, heavy sweating, prolonged thirst or
diarrhoea, is a stimulus for intensified release of the hormone.
Conversely, the secretion of vasopressin is inhibited by an
increase in the intravascular volume, e.g. as a result of increased
fluid intake.
[0004] Vasopressin exerts its action mainly via binding to three
receptors, which are classified as V1a, V1b and V2 receptors and
which belong to the family of G protein-coupled receptors. V1a
receptors are mainly located on the cells of the vascular smooth
musculature. Their activation gives rise to vasoconstriction, as a
result of which the peripheral resistance and blood pressure rise.
Apart from this, V1a receptors are also detectable in the liver.
V1b receptors (also named V3 receptors) are detectable in the
central nervous system. Together with corticotropin-releasing
hormone (CRH), vasopressin regulates the basal and stress-induced
secretion of adrenocorticotropic hormone (ACTH) via the V1b
receptor. V2 receptors are located in the distal tubular epithelium
and the epithelium of the collecting tubules in the kidney. Their
activation renders these epithelia permeable to water. This
phenomenon is due to the incorporation of aquaporins (special water
channels) in the luminal membrane of the epithelial cells.
[0005] The importance of vasopressin for the reabsorption of water
from the urine in the kidney becomes clear from the clinical
picture of diabetes insipidus, which is caused by a deficiency of
the hormone, e.g. owing to hypophysis damage. Patients who suffer
from this disease excrete up to 20 liters of urine per 24 hours if
they are not given replacement hormone. This volume corresponds to
about 10% of the primary urine. Because of its great importance for
the reabsorption of water from the urine, vasopressin is also
synonymously referred to as antidiuretic hormone (ADH).
Consequently, pharmacological inhibition of the action of
vasopressin/ADH on the V2 receptor results in increased urine
excretion. In contrast to the action of other diuretics (thiazides
and loop diuretics), however, V2 receptor antagonists cause
increased water excretion, without substantially increasing the
excretion of electrolytes. This means that with V2 antagonist
drugs, volume homeostasis can be restored without affecting
electrolyte homeostasis. Hence, drugs with V2 antagonistic activity
appear particularly suitable for the treatment of all disease
conditions which are associated with an overloading of the body
with water, without the electrolytes being adequately increased in
parallel.
[0006] A significant electrolyte abnormality is measurable in
clinical chemistry as hyponatremia (sodium concentration <135
mmol/L); it is the most important electrolyte abnormality in
hospital patients, with an incidence of about 5% or 250 000 cases
per year in the US alone. If the plasma sodium concentration falls
below 115 mmol/L, comatose states and death are imminent. Depending
on the underlying cause, a distinction is made between hypovolemic,
euvolemic and hypervolemic hyponatremia. The forms of hypervolemia
with edema formation are clinically significant. Typical examples
of these are the syndrome of inappropriate ADH/vasopressin
secretion (SIADH) (e.g. after craniocerebral trauma or as
paraneoplasia in carcinomas) and hypervolemic hyponatremia in liver
cirrhosis, various renal diseases and heart failure [De Luca L. et
al., Am. J. Cardiol. 96 (suppl.), 19L-23L (2005)]. In particular,
patients with heart failure, in spite of their relative
hyponatremia and hypervolemia, often display elevated vasopressin
levels, which are seen as the consequence of a generally disturbed
neurohumoral regulation in heart failure [Francis G. S. et al.,
Circulation 82, 1724-1729 (1990)].
[0007] The disturbed neurohormonal regulation essentially manifests
itself in an elevation of the sympathetic tone and inappropriate
activation of the renin-angiotensin-aldosterone system. While the
inhibition of these components by beta-receptor blockers on the one
hand and by ACE inhibitors or angiotensin-receptor blockers on the
other is now an inherent part of the pharmacological treatment of
heart failure, the inappropriate elevation of vasopressin secretion
in advanced heart failure is at present still not adequately
treatable. Apart from the retention of water mediated by V2
receptors and the unfavourable hemodynamic consequences associated
therewith in terms of increased backload, the emptying of the left
ventricle, the pressure in the pulmonary blood vessels and cardiac
output are also adversely affected by V1a-mediated
vasoconstriction. Furthermore, on the basis of experimental data in
animals, a direct hypertrophy-promoting action on the heart muscle
is also attributed to vasopressin. In contrast to the renal effect
of volume expansion, which is mediated by activation of V2
receptors, the direct action on the heart muscle is triggered by
activation of V1a receptors.
[0008] For these reasons, agents which inhibit the action of
vasopressin on the V2 and/or the V1a receptor appear suitable for
the treatment of heart failure. In particular, compounds with
combined activity on both vasopressin receptors (V1a and V2) should
have both desirable renal as well as hemodynamic effects and thus
offer an especially ideal profile for the treatment of patients
with heart failure. The provision of such combined vasopressin
antagonists also appears to make sense inasmuch as a volume
diminution mediated solely via V2 receptor blockade can entail the
stimulation of osmoreceptors and, as a result, may lead to a
further compensatory increase in vasopressin release. Through this,
in the absence of a component simultaneously blocking the V1a
receptor, the harmful effects of vasopressin, such as for example
vasoconstriction and heart muscle hypertrophy, could be further
intensified [Saghi P. et al., Europ. Heart J. 26, 538-543
(2005)].
[0009] Certain 4-phenyl-1,2,4-triazol-3-yl derivatives have been
described in WO 2005/063754-A1 and WO 2005/105779-A1 to act as
vasopressin V1a receptor antagonists that are useful for the
treatment of gynecological disorders, notably menstrual disorders
such as dysmenorrhea.
[0010] In WO 2011/104322-A1, a particular group of bis-aryl-bonded
1,2,4-triazol-3-ones, including 5-phenyl-1,2,4-triazol-3-yl and
1-phenyl-1,2,3-triazol-4-yl derivatives thereof, has been disclosed
as antagonists of vasopressin V1a and/or V2 receptors being useful
for the treatment and/or prevention of cardiovascular diseases.
During further investigation of this structural class it emerged,
however, that candidate compounds were frequently compromised by an
unsatisfactory aquaretic potency when evaluated in vivo following
peroral administration to conscious rats. Yet, as outlined above, a
robust aquaretic efficacy is a desirable prerequisite for the
treatment of disease conditions that are associated with an
overloading of the body with water, such as, for example, in
congestive heart failure.
[0011] A significant increase in aquaretic potency would also help
towards reducing the amount of substance which is going to be
required to achieve and maintain the desired therapeutic effect,
thus limiting the potential for unacceptable side effects and/or
unwanted drug-drug interactions during the treatment of patients
which might already be at high risk, such as, for example, in acute
or chronic heart failure or renal failure.
[0012] The technical problem to be solved according to the present
invention may therefore be seen in identifying and providing new
compounds that act as potent antagonists of both vasopressin V1a
and V2 receptors and, in addition, exhibit a substantial increase
in aquaretic potency in vivo.
[0013] Surprisingly, it has now been found that certain
5-(hydroxyalkyl)-1-phenyl-1,2,4-triazole derivatives represent
highly potent dual antagonists of vasopressin V1a and V2 receptors
exhibiting significantly enhanced aquaretic potency in vivo after
oral application. This improved activity profile renders the
compounds of the present invention particularly useful for the
treatment and/or prevention of cardiovascular and renal
diseases.
[0014] In one aspect, the present invention relates to
5-(hydroxyalkyl)-1-phenyl-1,2,4-triazole derivatives of the general
formula (I)
##STR00001##
wherein [0015] R.sup.1 is hydrogen or methyl, [0016] and [0017]
R.sup.2A and R.sup.2B are independently selected from the group
consisting of hydrogen, fluoro, chloro, cyano, methyl,
fluoromethyl, difluoromethyl, trifluoromethyl, ethyl, methoxy,
difluoromethoxy and trifluoromethoxy.
[0018] The compounds according to this invention can also be
present in the form of their salts, solvates and/or solvates of the
salts.
[0019] Compounds according to the invention are the compounds of
the formula (I) and their salts, solvates and solvates of the
salts, the compounds included in the formula (I) of the formulae
mentioned in the following and their salts, solvates and solvates
of the salts, and the compounds included in the formula (I) and
mentioned in the following as process products and/or embodiment
examples and their salts, solvates and solvates of the salts, where
the compounds included in the formula (I) and mentioned in the
following are not already salts, solvates and solvates of the
salts.
[0020] Salts for the purposes of the present invention are
preferably pharmaceutically acceptable salts of the compounds
according to the invention (for example, see S. M. Berge et al.,
"Pharmaceutical Salts", J. Pharm. Sci. 1977, 66, 1-19). Salts which
are not themselves suitable for pharmaceutical uses but can be
used, for example, for isolation, purification or storage of the
compounds according to the invention are also included.
[0021] Pharmaceutically acceptable salts 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,
benzenesulfonic acid, toluenesulfonic acid, naphthalenedisulfonic
acid, formic acid, acetic acid, trifluoroacetic acid, propionic
acid, lactic acid, tartaric acid, malic acid, citric acid, fumaric
acid, maleic acid, and benzoic acid.
[0022] Pharmaceutically acceptable salts also include salts of
customary bases, such as for example alkali metal salts (for
example sodium and potassium salts), alkaline earth metal salts
(for example calcium and magnesium salts), and ammonium salts
derived from ammonia or organic amines, such as illustratively and
preferably ethylamine, diethylamine, triethylamine,
N,N-diisopropylethylamine, monoethanolamine, diethanolamine,
triethanolamine, dimethylaminoethanol, diethylaminoethanol,
procaine, dicyclohexylamine, dibenzylamine, N-methylmorpholine,
N-methylpiperidine, arginine, lysine, and 1,2-ethylenediamine.
[0023] Solvates in the context of the invention are designated as
those forms of the compounds according to the invention which form
a complex in the solid or liquid state by stoichiometric
coordination with solvent molecules. Hydrates are a specific form
of solvates, in which the coordination takes place with water.
Hydrates are preferred solvates in the context of the present
invention.
[0024] The compounds of this invention may, either by nature of
asymmetric centers or by restricted rotation, be present in the
form of isomers (enantiomers, diastereomers). Any isomer may be
present in which the asymmetric center is in the (R)-, (S)-, or
(R,S)-configuration.
[0025] It will also be appreciated that when two or more asymmetric
centers are present in the compounds of the invention, several
diastereomers and enantiomers of the exemplified structures will
often be possible, and that pure diastereomers and pure enantiomers
represent preferred embodiments. It is intended that pure
stereoisomers, pure diastereomers, pure enantiomers, and mixtures
thereof, are within the scope of the invention.
[0026] All isomers, whether separated, pure, partially pure, or in
racemic mixture, of the compounds of this invention are encompassed
within the scope of this invention. The purification of said
isomers and the separation of said isomeric mixtures may be
accomplished by standard techniques known in the art. For example,
diastereomeric mixtures can be separated into the individual
isomers by chromatographic processes or crystallization, and
racemates can be separated into the respective enantiomers either
by chromatographic processes on chiral phases or by resolution.
[0027] In addition, all possible tautomeric forms of the compounds
described above are included according to the present
invention.
[0028] The present invention also encompasses all suitable isotopic
variants of the compounds according to the invention. An isotopic
variant of a compound according to the invention is understood to
mean a compound in which at least one atom within the compound
according to the invention 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, fluorine, chlorine, bromine and iodine, such as .sup.2H
(deuterium), .sup.3H (tritium), .sup.13C, .sup.14C, .sup.15N,
.sup.17O, .sup.18O, .sup.18F, .sup.36Cl, .sup.82Br, .sup.123I,
.sup.124I, .sup.129I and .sup.131I. 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 compound distribution in the body. Due to
comparatively easy preparability and detectability, especially
compounds labelled with .sup.3H, .sup.14C and/or .sup.18F isotopes
are suitable for this purpose. In addition, the incorporation of
isotopes, for example of deuterium, can 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 compounds according to the invention 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 processes known to those
skilled in the art, for example by the methods described below and
the methods described in the working examples, by using
corresponding isotopic modifications of the particular reagents
and/or starting compounds therein.
[0029] In a distinct embodiment, the present invention relates to
compounds of formula (I), wherein R.sup.1 is methyl.
[0030] In a further distinct embodiment, the present invention
relates to compounds of formula (I), wherein at least one of
R.sup.2A and R.sup.2B is other than hydrogen.
[0031] In another distinct embodiment, the present invention
relates to compounds of formula (I), wherein [0032] R.sup.1 is
hydrogen or methyl, [0033] and [0034] R.sup.2A and R.sup.2B are
independently selected from the group consisting of hydrogen,
fluoro, chloro, methyl and methoxy, wherein at least one of
R.sup.2A and R.sup.2B is other than hydrogen.
[0035] In a preferred embodiment, the present invention relates to
compounds according to formula (I) selected from the group
consisting of the following compounds [0036]
5-(4-chlorophenyl)-2-{[1-(3-chlorophenyl)-5-(hydroxymethyl)-1H-1,2,4-tria-
zol-3-yl]methyl}-4-[(2S)-3,3,3-trifluoro-2-hydroxypropyl]-2,4-dihydro-3H-1-
,2,4-triazol-3-one; [0037]
5-(4-chlorophenyl)-2-{[1-(3-fluorophenyl)-5-(hydroxymethyl)-1H-1,2,4-tria-
zol-3-yl]methyl}-4-[(2S)-3,3,3-trifluoro-2-hydroxypropyl]-2,4-dihydro-3H-1-
,2,4-triazol-3-one; [0038]
5-(4-chlorophenyl)-2-{[5-(hydroxymethyl)-1-(2-methylphenyl)-1H-1,2,4-tria-
zol-3-yl]methyl}-4-[(2S)-3,3,3-trifluoro-2-hydroxypropyl]-2,4-dihydro-3H-1-
,2,4-triazol-3-one; [0039]
2-({1-(2-chloro-4-fluorophenyl)-5-[(1RS)-1-hydroxyethyl]-1H-1,2,4-triazol-
-3-yl}methyl)-5-(4-chlorophenyl)-4-[(2S)-3,3,3-trifluoro-2-hydroxypropyl]--
2,4-dihydro-3H-1,2,4-triazol-3-one; [0040]
2-{[1-(2-chloro-4-fluorophenyl)-5-(1-hydroxyethyl)-1H-1,2,4-triazol-3-yl]-
methyl}-5-(4-chlorophenyl)-4-[(2S)-3,3,3-trifluoro-2-hydroxypropyl]-2,4-di-
hydro-3H-1,2,4-triazol-3-one (diastereomer 1); [0041]
2-{[1-(2-chloro-4-fluorophenyl)-5-(1-hydroxyethyl)-1H-1,2,4-triazol-3-yl]-
methyl}-5-(4-chlorophenyl)-4-[(2S)-3,3,3-trifluoro-2-hydroxypropyl]-2,4-di-
hydro-3H-1,2,4-triazol-3-one (diastereomer 2); [0042]
2-({1-(2-chloro-5-fluorophenyl)-5-[(1RS)-1-hydroxyethyl]-1H-1,2,4-triazol-
-3-yl}methyl)-5-(4-chlorophenyl)-4-[(2S)-3,3,3-trifluoro-2-hydroxypropyl]--
2,4-dihydro-3H-1,2,4-triazol-3-one; [0043]
2-{[1-(2-chloro-5-fluorophenyl)-5-(1-hydroxyethyl)-1H-1,2,4-triazol-3-yl]-
methyl}-5-(4-chlorophenyl)-4-[(2S)-3,3,3-trifluoro-2-hydroxypropyl]-2,4-di-
hydro-3H-1,2,4-triazol-3-one (diastereomer 1); [0044]
2-{[1-(2-chloro-5-fluorophenyl)-5-(1-hydroxyethyl)-1H-1,2,4-triazol-3-yl]-
methyl}-5-(4-chlorophenyl)-4-[(2S)-3,3,3-trifluoro-2-hydroxypropyl]-2,4-di-
hydro-3H-1,2,4-triazol-3-one (diastereomer 2); [0045]
5-(4-chlorophenyl)-2-({1-(3-fluorophenyl)-5-[(1RS)-1-hydroxyethyl]-1H-1,2-
,4-triazol-3-yl}-methyl)-4-[(2S)-3,3,3-trifluoro-2-hydroxypropyl]-2,4-dihy-
dro-3H-1,2,4-triazol-3-one; [0046]
5-(4-chlorophenyl)-2-({1-(3-fluorophenyl)-5-[(1R)-1-hydroxyethyl]-1H-1,2,-
4-triazol-3-yl}methyl)-4-[(2S)-3,3,3-trifluoro-2-hydroxypropyl]-2,4-dihydr-
o-3H-1,2,4-triazol-3-one; [0047]
5-(4-chlorophenyl)-2-({1-(3-fluorophenyl)-5-[(1S)-1-hydroxyethyl]-1H-1,2,-
4-triazol-3-yl}methyl)-4-[(2S)-3,3,3-trifluoro-2-hydroxypropyl]-2,4-dihydr-
o-3H-1,2,4-triazol-3-one; [0048]
5-(4-chlorophenyl)-2-({1-(3-chlorophenyl)-5-[(1RS)-1-hydroxyethyl]-1H-1,2-
,4-triazol-3-yl}-methyl)-4-[(2S)-3,3,3-trifluoro-2-hydroxypropyl]-2,4-dihy-
dro-3H-1,2,4-triazol-3-one; [0049]
5-(4-chlorophenyl)-2-({1-(3-chlorophenyl)-5-[(1R)-1-hydroxyethyl]-1H-1,2,-
4-triazol-3-yl}methyl)-4-[(2S)-3,3,3-trifluoro-2-hydroxypropyl]-2,4-dihydr-
o-3H-1,2,4-triazol-3-one; [0050]
5-(4-chlorophenyl)-2-({1-(3-chlorophenyl)-5-[(1S)-1-hydroxyethyl]-1H-1,2,-
4-triazol-3-yl}methyl)-4-[(2S)-3,3,3-trifluoro-2-hydroxypropyl]-2,4-dihydr-
o-3H-1,2,4-triazol-3-one; [0051]
5-(4-chlorophenyl)-2-({1-(2-chlorophenyl)-5-[(1RS)-1-hydroxyethyl]-1H-1,2-
,4-triazol-3-yl}-methyl)-4-[(2S)-3,3,3-trifluoro-2-hydroxypropyl]-2,4-dihy-
dro-3H-1,2,4-triazol-3-one; [0052]
5-(4-chlorophenyl)-2-({1-(2-chlorophenyl)-5-[(1R)-1-hydroxyethyl]-1H-1,2,-
4-triazol-3-yl}methyl)-4-[(2S)-3,3,3-trifluoro-2-hydroxypropyl]-2,4-dihydr-
o-3H-1,2,4-triazol-3-one; [0053] and [0054]
5-(4-chlorophenyl)-2-({1-(2-chlorophenyl)-5-[(1S)-1-hydroxyethyl]-1H-1,2,-
4-triazol-3-yl}methyl)-4-[(2S)-3,3,3-trifluoro-2-hydroxypropyl]-2,4-dihydr-
o-3H-1,2,4-triazol-3-one.
[0055] In a particularly preferred embodiment, the present
invention relates to compounds according to formula (I) selected
from the group consisting of the following compounds [0056]
5-(4-chlorophenyl)-2-({1-(3-fluorophenyl)-5-[(1RS)-1-hydroxyethyl]-1H-1,2-
,4-triazol-3-yl}-methyl)-4-[(2S)-3,3,3-trifluoro-2-hydroxypropyl]-2,4-dihy-
dro-3H-1,2,4-triazol-3-one; [0057]
5-(4-chlorophenyl)-2-({1-(3-fluorophenyl)-5-[(1S)-1-hydroxyethyl]-1H-1,2,-
4-triazol-3-yl}methyl)-4-[(2S)-3,3,3-trifluoro-2-hydroxypropyl]-2,4-dihydr-
o-3H-1,2,4-triazol-3-one; [0058]
5-(4-chlorophenyl)-2-({1-(3-chlorophenyl)-5-[(1RS)-1-hydroxyethyl]-1H-1,2-
,4-triazol-3-yl}-methyl)-4-[(2S)-3,3,3-trifluoro-2-hydroxypropyl]-2,4-dihy-
dro-3H-1,2,4-triazol-3-one; [0059]
5-(4-chlorophenyl)-2-({1-(3-chlorophenyl)-5-[(1S)-1-hydroxyethyl]-1H-1,2,-
4-triazol-3-yl}methyl)-4-[(2S)-3,3,3-trifluoro-2-hydroxypropyl]-2,4-dihydr-
o-3H-1,2,4-triazol-3-one; [0060]
5-(4-chlorophenyl)-2-({1-(2-chlorophenyl)-5-[(1RS)-1-hydroxyethyl]-1H-1,2-
,4-triazol-3-yl}-methyl)-4-[(2S)-3,3,3-trifluoro-2-hydroxypropyl]-2,4-dihy-
dro-3H-1,2,4-triazol-3-one; [0061] and [0062]
5-(4-chlorophenyl)-2-({1-(2-chlorophenyl)-5-[(1S)-1-hydroxyethyl]-1H-1,2,-
4-triazol-3-yl}methyl)-4-[(2S)-3,3,3-trifluoro-2-hydroxypropyl]-2,4-dihydr-
o-3H-1,2,4-triazol-3-one.
[0063] In a further embodiment, the present invention relates to a
process for preparing the compounds of the general formula (I),
characterized in that a compound of formula (II)
##STR00002##
is first reacted with hydrazine to give the hydrazide of formula
(III)
##STR00003##
then condensed with an amidine of formula (IV)
##STR00004##
or a salt thereof, wherein R.sup.1 has the meaning described above,
in the presence of a base to give a 1,2,4-triazole derivative of
formula (V)
##STR00005##
and/or a tautomer thereof, wherein R.sup.1 has the meaning
described above, and subsequently coupled with a phenylboronic acid
of formula (VI)
##STR00006##
wherein R.sup.2A and R.sup.2B have the meanings described above, in
the presence of a copper catalyst and an amine base to yield the
target compound of formula (I)
##STR00007##
wherein R.sup.1, R.sup.2A and R.sup.2B have the meanings described
above, optionally followed, where appropriate, by (i) separating
the compounds of formula (I) thus obtained into their respective
diastereomers, preferably using chromatographic methods, and/or
(ii) converting the compounds of formula (I) into their respective
hydrates, solvates, salts and/or hydrates or solvates of the salts
by treatment with the corresponding solvents and/or acids or
bases.
[0064] Compounds of formula (I), wherein R.sup.1 represents methyl,
can also be obtained in diastereomerically pure form by employing
the appropriate enantiomer of amidine (IV) [R.sup.1=methyl], i.e.
(IV-A) or (IV-B)
##STR00008##
or a salt thereof, in the condensation reaction described
above.
[0065] The transformation (II).fwdarw.(III) is carried out in the
usual way by treating methyl ester (II) with hydrazine or hydrazine
hydrate in an alcoholic solvent, such as methanol, ethanol,
n-propanol, isopropanol or n-butanol, at a temperature in the range
of +20.degree. C. to +100.degree. C.
[0066] The condensation reaction (III)+(IV).fwdarw.(V) is usually
carried out in an inert dipolar-aprotic solvent, such as
N,N-dimethylformamide (DMF), N,N-dimethylacetamide (DMA),
dimethylsulfoxide (DMSO), N-methylpyrrolidinone (NMP) or
N,N'-dimethylpropylene urea (DMPU), in the presence of a
sufficiently strong base, such as sodium hydride or a sodium or
potassium alkoxide, for example sodium or potassium methoxide,
sodium or potassium ethoxide, or sodium or potassium tert-butoxide.
The amidine (IV) may be employed as such in this reaction or in
salt form, e.g. as the hydrochloride salt. In the latter case, a
proportional excess of base is used. The reaction is generally
performed at a temperature between +80.degree. C. and +150.degree.
C. Heating by means of a microwave reactor device may have a
beneficial effect for this condensation reaction.
[0067] The 1,2,4-triazole derivative of formula (V) produced by
this reaction may also be present in other tautomeric forms, such
as (V-A) or (V-B)
##STR00009##
or as a mixture of tautomers.
[0068] The coupling reaction (V)+(VI).fwdarw.(I) is typically
carried out with the aid of a copper catalyst and an amine base
["Chan-Lam coupling" conditions; see, for instance, D. M. T. Chan
et al., Tetrahedron Lett. 44 (19), 3863-3865 (2003); J. X. Qiao and
P. Y. S. Lam, Synthesis, 829-856 (2011); K. S. Rao and T.-S. Wu,
Tetrahedron 68, 7735-7754 (2012)]. Copper catalysts suitable for
this process are in particular copper(II) salts, such as copper(II)
acetate, copper(II) trifluoromethanesulfonate or copper(II)
bromide. Practical amine bases include, for example, triethylamine,
N,N-diisopropylethylamine, pyridine and
4-(N,N-dimethylamino)pyridine. The reaction is performed in an
inert organic solvent, such as dichloromethane, 1,2-dichloroethane,
methyl tert-butyl ether, tetrahydrofuran, 1,4-dioxane,
1,2-dimethoxyethane, toluene, pyridine, ethyl acetate, acetonitrile
or N,N-dimethylformamide, or in a mixture of these solvents.
Preferably, pyridine is used both as solvent and base. The coupling
is generally carried out at a temperature in the range of
+20.degree. C. to +120.degree. C., preferably at +20.degree. C. to
+70.degree. C. Concomitant microwave irradiation may have a
beneficial effect in this reaction as well.
[0069] Regioisomeric phenyltriazole derivatives which may arise
from a coupling reaction occurring at other triazole nitrogen atoms
[cf tautomers (V-A), (V-B)] can, in the event, be readily separated
from the target product (I) by conventional HPLC
chromatography.
[0070] The compound of formula (II) can be synthesized by the
procedures described in Int. Pat. Appl. WO 2011/104322-A1 (see also
synthesis schemes 1a and 1b below).
[0071] The compounds of the formulae (IV), (IV-A), (IV-B) and (VI)
are either commercially available, known from the literature, or
can be prepared from readily available starting materials by
adaptation of standard methods described in the literature.
Detailed procedures and literature references for preparing the
starting materials can also be found in the Experimental Part in
the section on the preparation of the starting materials and
intermediates.
[0072] The preparation of the compounds of the invention may be
illustrated by means of the following synthesis schemes:
##STR00010##
##STR00011##
##STR00012##
[0073] The compounds of the present invention have valuable
pharmacological properties and can be used for the prevention
and/or treatment of various diseases and disease-induced states in
humans and other mammals.
[0074] In the context of the present invention, the term
"treatment" or "treating" includes inhibiting, delaying, relieving,
mitigating, arresting, reducing, or causing the regression of a
disease, disorder, condition, or state, the development and/or
progression thereof, and/or the symptoms thereof. The term
"prevention" or "preventing" includes reducing the risk of having,
contracting, or experiencing a disease, disorder, condition, or
state, the development and/or progression thereof, and/or the
symptoms thereof. The term prevention includes prophylaxis.
Treatment or prevention of a disorder, disease, condition, or state
may be partial or complete.
[0075] Throughout this document, for the sake of simplicity, the
use of singular language is given preference over plural language,
but is generally meant to include the plural language if not
otherwise stated. For example, the expression "A method of treating
a disease in a patient, comprising administering to a patient an
effective amount of a compound of formula (I)" is meant to include
the simultaneous treatment of more than one disease as well as the
administration of more than one compound of formula (I).
[0076] The compounds of the present invention are highly potent
dual antagonists of vasopressin V1a and V2 receptors. In addition,
the compounds of the invention exhibit a pronounced aquaretic
effect in vivo after oral application. The compounds of the
invention are therefore expected to be highly valuable as
therapeutic agents for the treatment and/or prevention of diseases,
in particular for the treatment and/or prevention of cardiovascular
and renal diseases.
[0077] Cardiovascular diseases in this context that may be treated
and/or prevented with the compounds of the invention include, but
are not limited to, the following: acute and chronic heart failure
including worsening chronic heart failure (or hospitalization for
heart failure) and congestive heart failure, arterial hypertension,
resistant hypertension, arterial pulmonary hypertension, coronary
heart disease, stable and unstable angina pectoris, atrial and
ventricular arrhythmias, disturbances of atrial and ventricular
rhythm and conduction disturbances, for example atrioventricular
blocks of degree I-III (AVB I-III), supraventricular
tachyarrhythmia, atrial fibrillation, atrial flutter, ventricular
fibrillation, ventricular flutter, ventricular tachyarrhythmia,
torsade-de-pointes tachycardia, atrial and ventricular
extrasystoles, AV-junction extrasystoles, sick-sinus syndrome,
syncopes, AV-node re-entry tachycardia and Wolff-Parkinson-White
syndrome, acute coronary syndrome (ACS), autoimmune heart diseases
(pericarditis, endocarditis, valvulitis, aortitis,
cardiomyopathies), shock such as cardiogenic shock, septic shock
and anaphylactic shock, aneurysms, Boxer cardiomyopathy (premature
ventricular contraction), furthermore thromboembolic diseases and
ischaemias such as peripheral perfusion disturbances, reperfusion
injury, arterial and venous thromboses, myocardial insufficiency,
endothelial dysfunction, micro- and macrovascular damage
(vasculitis) and for preventing restenoses such as after
thrombolysis therapies, percutaneous transluminal angioplasty
(PTA), percutaneous transluminal coronary angioplasty (PTCA), heart
transplantation and bypass operations, arteriosclerosis,
disturbances of lipid metabolism, hypolipoproteinaemias,
dyslipidaemias, hypertriglyceridaemias, hyperlipidaemias and
combined hyperlipidaemias, hypercholesterolaemias,
abetalipoproteinaemia, sitosterolaemia, xanthomatosis, Tangier
disease, adipositas, obesity, metabolic syndrome, transitory and
ischaemic attacks, stroke, inflammatory cardiovascular diseases,
peripheral and cardiac vascular diseases, peripheral circulation
disorders, spasms of the coronary arteries and peripheral arteries,
and edema such as, for example, pulmonary edema, cerebral edema,
renal edema and heart failure-related edema.
[0078] In the sense of the present invention, the term heart
failure also includes more specific or related disease forms such
as right heart failure, left heart failure, global insufficiency,
ischaemic cardiomyopathy, dilatative cardiomyopathy, congenital
heart defects, heart valve defects, heart failure with heart valve
defects, mitral valve stenosis, mitral valve insufficiency, aortic
valve stenosis, aortic valve insufficiency, tricuspidal stenosis,
tricuspidal insufficiency, pulmonary valve stenosis, pulmonary
valve insufficiency, combined heart valve defects, heart muscle
inflammation (myocarditis), chronic myocarditis, acute myocarditis,
viral myocarditis, diabetic heart failure, alcohol-toxic
cardiomyopathy, cardiac storage diseases, heart failure with
preserved ejection fraction (HFpEF or diastolic heart failure), and
heart failure with reduced ejection fraction (HFrEF or systolic
heart failure).
[0079] The compounds according to the invention are also suitable
for the treatment and/or prevention of renal diseases, in
particular of acute and chronic renal insufficiency, and of acute
and chronic renal failure. In the sense of the present invention,
the term renal insufficiency comprises both acute and chronic
manifestations of renal insufficiency, as well as underlying or
related kidney diseases such as renal hypoperfusion, intradialytic
hypotension, obstructive uropathy, glomerulopathies,
glomerulonephritis, acute glomerulonephritis, glomerulosclerosis,
tubulointerstitial diseases, nephropathic diseases such as primary
and congenital kidney disease, nephritis, immunological kidney
diseases such as kidney transplant rejection, immune
complex-induced kidney diseases, nephropathy induced by toxic
substances, contrast medium-induced nephropathy, diabetic and
non-diabetic nephropathy, pyelonephritis, renal cysts,
nephrosclerosis, hypertensive nephrosclerosis and nephrotic
syndrome, which can be characterized diagnostically, for example,
by abnormally reduced creatinine and/or water excretion, abnormally
increased blood concentrations of urea, nitrogen, potassium and/or
creatinine, altered activity of renal enzymes such as, for example,
glutamyl synthetase, altered urine osmolarity or urine volume,
increased microalbuminuria, macroalbuminuria, lesions of glomeruli
and arterioles, tubular dilatation, hyperphosphataemia and/or the
need for dialysis. The present invention also comprises the use of
the compounds according to the invention for the treatment and/or
prevention of sequelae of renal insufficiency, for example
pulmonary edema, heart failure, uraemia, anaemia, electrolyte
disturbances (e.g. hyperkalaemia, hyponatraemia) and disturbances
in bone and carbohydrate metabolism.
[0080] The compounds of the present invention may be particularly
useful for the treatment and/or prevention of the cardiorenal
syndrome (CRS) and its various subtypes. This term embraces certain
disorders of the heart and kidneys whereby acute or chronic
dysfunction in one organ may induce acute or chronic dysfunction of
the other. CRS has been sub-classified into five types based upon
the organ that initiated the insult as well as the acuity and
chronicity of the disease (type 1: development of renal
insufficiency resulting from acute decompensated heart failure;
type 2: chronic congestive heart failure resulting in progressive
renal dysfunction; type 3: acute cardiac dysfunction resulting from
an abrupt fall in renal function; type 4: chronic kidney disease
leading to cardiac remodeling; type 5: systemic disease involving
both the heart and the kidneys) [see, for example, M. R. Kahn et
al., Nature Rev. Cardiol. 10, 261-273 (2013)].
[0081] The compounds according to the invention are also suitable
for the treatment and/or prevention of polycystic kidney disease
(PCKD) and of the syndrome of inadequate ADH secretion (SIADH).
Furthermore, the compounds of the invention are suitable for use as
a diuretic for the treatment of edemas and in electrolyte
disorders, in particular in hypervolemic and euvolemic
hyponatremia.
[0082] Moreover, the compounds according to the invention may be
used for the treatment and/or prevention of primary and secondary
Raynaud's phenomenon, microcirculation disturbances, claudication,
peripheral and autonomic neuropathies, diabetic microangiopathies,
diabetic retinopathy, diabetic limb ulcers, gangrene, CREST
syndrome, erythematous disorders, onychomycosis, rheumatic diseases
and for promoting wound healing.
[0083] Furthermore, the compounds of the invention are suitable for
treating urological diseases and diseases of the male and female
urogenital system such as, for example, benign prostatic syndrome
(BPS), benign prostatic hyperplasia (BPH), benign prostatic
enlargement (BPE), bladder outlet obstruction (BOO), lower urinary
tract syndromes (LUTS), neurogenic overactive bladder (OAB),
interstitial cystitis (IC), urinary incontinence (UI), for example
mixed, urge, stress and overflow incontinence (MUI, UUI, SUI, OUI),
pelvic pains, erectile dysfunction and female sexual
dysfunction.
[0084] The compounds according to the invention may also be used
for the treatment and/or prevention of inflammatory diseases,
asthmatic diseases, chronic obstructive pulmonary disease (COPD),
acute respiratory distress syndrome (ARDS), acute lung injury
(ALI), alpha-1-antitrypsin deficiency (AATD), pulmonary fibrosis,
pulmonary emphysema (e.g. smoking-induced pulmonary emphysema) and
cystic fibrosis (CF). In addition, the compounds of the invention
may be used for the treatment and/or prevention of pulmonary
arterial hypertension (PAH) and other forms of pulmonary
hypertension (PH), including pulmonary hypertension associated with
left ventricular disease, HIV infection, sickle cell anaemia,
thromboembolism (CTEPH), sarcoidosis, chronic obstructive pulmonary
disease (COPD) or pulmonary fibrosis.
[0085] Additionally, the compounds according to the invention may
be used for the treatment and/or prevention of liver cirrhosis,
ascites, diabetes mellitus and diabetic complications such as, for
example, neuropathy and nephropathy. Further, the compounds of the
invention are suitable for the treatment and/or prevention of
central nervous disorders such as anxiety states and depression, of
glaucoma and of cancer, in particular of pulmonary tumors, and for
the management of circadian rhythm misalignment such as jet lag and
shift work.
[0086] Furthermore, the compounds according to the invention may be
useful for the treatment and/or prevention of pain conditions,
diseases of the adrenals such as, for example, pheochromocytoma and
adrenal apoplexy, diseases of the intestine such as, for example,
Crohn's disease and diarrhea, menstrual disorders such as, for
example, dysmenorrhea, or of endometriosis, preterm labour and for
tocolysis.
[0087] Due to their activity and selectivity profile, the compounds
of the present invention are believed to be particularly suitable
for the treatment and/or prevention of acute and chronic heart
failure, cardiorenal syndrome (type 1-5), hypervolemic and
euvolemic hyponatremia, liver cirrhosis, ascites, edema and the
syndrome of inadequate ADH secretion (SIADH).
[0088] The diseases mentioned above have been well characterized in
humans, but also exist with a comparable aetiology in other
mammals, and may be treated in those with the compounds and methods
of the present invention.
[0089] Thus, the present invention further relates to the use of
the compounds according to the invention for the treatment and/or
prevention of diseases, especially of the aforementioned
diseases.
[0090] The present invention further relates to the use of the
compounds according to the invention for preparing a pharmaceutical
composition for the treatment and/or prevention of diseases,
especially of the aforementioned diseases.
[0091] The present invention further relates to the use of the
compounds according to the invention in a method for the treatment
and/or prevention of diseases, especially of the aforementioned
diseases.
[0092] The present invention further relates to a method for the
treatment and/or prevention of diseases, especially of the
aforementioned diseases, by using an effective amount of at least
one of the compounds according to the invention.
[0093] Compounds of the present invention may be administered as
the sole pharmaceutical agent or in combination with one or more
additional therapeutic agents as long as this combination does not
lead to undesirable and/or unacceptable side effects. Such
combination therapy includes administration of a single
pharmaceutical dosage formulation which contains a compound of
formula (I), as defined above, and one or more additional
therapeutic agents, as well as administration of a compound of
formula (I) and each additional therapeutic agent in its own
separate pharmaceutical dosage formulation. For example, a compound
of formula (I) and a therapeutic agent may be administered to the
patient together in a single (fixed) oral dosage composition such
as a tablet or capsule, or each agent may be administered in
separate dosage formulations.
[0094] Where separate dosage formulations are used, the compound of
formula (I) and one or more additional therapeutic agents may be
administered at essentially the same time (i.e., concurrently) or
at separately staggered times (i.e., sequentially).
[0095] In particular, the compounds of the present invention may be
used in fixed or separate combination with [0096] organic nitrates
and NO-donors, for example sodium nitroprusside, nitroglycerin,
isosorbide mononitrate, isosorbide dinitrate, molsidomine or SIN-1,
and inhalational NO; [0097] compounds that inhibit the degradation
of cyclic guanosine monophosphate (cGMP), for example inhibitors of
phosphodiesterases (PDE) 1, 2 and/or 5, in particular PDE-5
inhibitors such as sildenafil, vardenafil, tadalafil, udenafil,
dasantafil, avanafil, mirodenafil or lodenafil; [0098]
positive-inotropic agents, such as for example cardiac glycosides
(digoxin) and beta-adrenergic and dopaminergic agonists such as
isoproterenol, adrenalin, noradrenalin, dopamine or dobutamine;
[0099] natriuretic peptides, such as for example atrial natriuretic
peptide (ANP, anaritide), B-type natriuretic peptide or brain
natriuretic peptide (BNP, nesiritide), C-type natriuretic peptide
(CNP) or urodilatin; [0100] calcium sensitizers, such as for
example and preferably levosimendan; [0101] NO- and
heme-independent activators of soluble guanylate cyclase (sGC),
such as in particular cinaciguat and also the compounds described
in WO 01/19355, WO 01/19776, WO 01/19778, WO 01/19780, WO 02/070462
and WO 02/070510; [0102] NO-independent, but heme-dependent
stimulators of guanylate cyclase (sGC), such as in particular
riociguat, vericiguat and also the compounds described in WO
00/06568, WO 00/06569, WO 02/42301, WO 03/095451, WO 2011/147809,
WO 2012/004258, WO 2012/028647 and WO 2012/059549; [0103]
inhibitors of human neutrophil elastase (HNE), such as for example
sivelestat or DX-890 (reltran); [0104] compounds inhibiting the
signal transduction cascade, in particular tyrosine and/or
serine/threonine kinase inhibitors, such as for example nintedanib,
dasatinib, nilotinib, bosutinib, regorafenib, sorafenib, sunitinib,
cediranib, axitinib, telatinib, imatinib, brivanib, pazopanib,
vatalanib, gefitinib, erlotinib, lapatinib, canertinib,
lestaurtinib, pelitinib, semaxanib or tandutinib; [0105] compounds
influencing the energy metabolism of the heart, such as for example
and preferably etomoxir, dichloroacetate, ranolazine or
trimetazidine, or full or partial adenosine A1 receptor agonists;
[0106] compounds influencing the heart rate, such as for example
and preferably ivabradine; [0107] cardiac myosin activators, such
as for example and preferably omecamtiv mecarbil (CK-1827452);
[0108] antithrombotic agents, for example and preferably from the
group of platelet aggregation inhibitors, anticoagulants and
profibrinolytic substances; [0109] blood pressure lowering agents,
for example and preferably from the group of calcium antagonists,
angiotensin AII antagonists, ACE inhibitors, vasopeptidase
inhibitors, endothelin antagonists, renin inhibitors,
alpha-blockers, beta-blockers, mineralocorticoid receptor
antagonists and diuretics; and/or [0110] fat metabolism altering
agents, for example and preferably from the group of thyroid
receptor agonists, cholesterol synthesis inhibitors, such as for
example and preferably HMG-CoA-reductase or squalene synthesis
inhibitors, ACAT inhibitors, CETP inhibitors, MTP inhibitors,
PPAR-alpha, PPAR-gamma and/or PPAR-delta agonists, cholesterol
absorption inhibitors, lipase inhibitors, polymeric bile acid
adsorbers, bile acid reabsorption inhibitors and lipoprotein(a)
antagonists.
[0111] Antithrombotic agents are preferably to be understood as
compounds from the group of platelet aggregation inhibitors,
anticoagulants and profibrinolytic substances.
[0112] In a preferred embodiment of the invention, the compounds
according to the invention are administered in combination with a
platelet aggregation inhibitor, for example and preferably aspirin,
clopidogrel, ticlopidine or dipyridamole.
[0113] In a preferred embodiment of the invention, the compounds
according to the invention are administered in combination with a
thrombin inhibitor, for example and preferably ximelagatran,
dabigatran, melagatran, bivalirudin or enoxaparin.
[0114] In a preferred embodiment of the invention, the compounds
according to the invention are administered in combination with a
GPIIb/IIIa antagonist, for example and preferably tirofiban or
abciximab.
[0115] In a preferred embodiment of the invention, the compounds
according to the invention are administered in combination with a
factor Xa inhibitor, for example and preferably rivaroxaban,
apixaban, otamixaban, fidexaban, razaxaban, fondaparinux,
idraparinux, DU-176b, PMD-3112, YM-150, KFA-1982, EMD-503982,
MCM-17, MLN-1021, DX 9065a, DPC 906, JTV 803, SSR-126512 or
SSR-128428.
[0116] In a preferred embodiment of the invention, the compounds
according to the invention are administered in combination with
heparin or a low molecular weight (LMW) heparin derivative.
[0117] In a preferred embodiment of the invention, the compounds
according to the invention are administered in combination with a
vitamin K antagonist, for example and preferably coumarin.
[0118] Blood pressure lowering agents are preferably to be
understood as compounds from the group of calcium antagonists,
angiotensin AII antagonists, ACE inhibitors, vasopeptidase
inhibitors, endothelin antagonists, renin inhibitors,
alpha-blockers, beta-blockers, mineralocorticoid receptor
antagonists and diuretics.
[0119] In a preferred embodiment of the invention, the compounds
according to the invention are administered in combination with a
calcium antagonist, for example and preferably nifedipine,
amlodipine, verapamil or diltiazem.
[0120] In a preferred embodiment of the invention, the compounds
according to the invention are administered in combination with an
alpha-1-receptor blocker, for example and preferably prazosin or
tamsulosin.
[0121] In a preferred embodiment of the invention, the compounds
according to the invention are administered in combination with a
beta-blocker, for example and preferably propranolol, atenolol,
timolol, pindolol, alprenolol, oxprenolol, penbutolol, bupranolol,
metipranolol, nadolol, mepindolol, carazolol, sotalol, metoprolol,
betaxolol, celiprolol, bisoprolol, carteolol, esmolol, labetalol,
carvedilol, adaprolol, landiolol, nebivolol, epanolol or
bucindolol.
[0122] In a preferred embodiment of the invention, the compounds
according to the invention are administered in combination with an
angiotensin AII receptor antagonist, for example and preferably
losartan, candesartan, valsartan, telmisartan, irbesartan,
olmesartan, eprosartan or azilsartan.
[0123] In a preferred embodiment of the invention, the compounds
according to the invention are administered in combination with a
vasopeptidase inhibitor or inhibitor of neutral endopeptidase
(NEP), such as for example and preferably sacubitril, omapatrilat
or AVE-7688.
[0124] In a preferred embodiment of the invention, the compounds
according to the invention are administered in combination with a
dual angiotensin AII receptor antagonist/NEP inhibitor (ARNI), for
example and preferably LCZ696.
[0125] In a preferred embodiment of the invention, the compounds
according to the invention are administered in combination with an
ACE inhibitor, for example and preferably enalapril, captopril,
lisinopril, ramipril, delapril, fosinopril, quinopril, perindopril
or trandopril.
[0126] In a preferred embodiment of the invention, the compounds
according to the invention are administered in combination with an
endothelin antagonist, for example and preferably bosentan,
darusentan, ambrisentan, tezosentan or sitaxsentan.
[0127] In a preferred embodiment of the invention, the compounds
according to the invention are administered in combination with a
renin inhibitor, for example and preferably aliskiren, SPP-600 or
SPP-800.
[0128] In a preferred embodiment of the invention, the compounds
according to the invention are administered in combination with a
mineralocorticoid receptor antagonist, for example and preferably
finerenone, spironolactone, canrenone, potassium canrenoate or
eplerenone.
[0129] In a preferred embodiment of the invention, the compounds
according to the invention are administered in combination with a
diuretic, such as for example and preferably furosemide,
bumetanide, piretanide, torsemide, bendroflumethiazide,
chlorothiazide, hydrochlorothiazide, xipamide, indapamide,
hydroflumethiazide, methyclothiazide, polythiazide,
trichloromethiazide, chlorothalidone, metolazone, quinethazone,
acetazolamide, dichlorophenamide, methazolamide, glycerine,
isosorbide, mannitol, amiloride or triamterene.
[0130] Fat metabolism altering agents are preferably to be
understood as compounds from the group of CETP inhibitors, thyroid
receptor agonists, cholesterol synthesis inhibitors such as
HMG-CoA-reductase or squalene synthesis inhibitors, ACAT
inhibitors, MTP inhibitors, PPAR-alpha, PPAR-gamma and/or
PPAR-delta agonists, cholesterol absorption inhibitors, polymeric
bile acid adsorbers, bile acid reabsorption inhibitors, lipase
inhibitors and lipoprotein(a) antagonists.
[0131] In a preferred embodiment of the invention, the compounds
according to the invention are administered in combination with a
CETP inhibitor, for example and preferably dalcetrapib,
anacetrapib, BAY 60-5521 or CETP-vaccine (Avant).
[0132] In a preferred embodiment of the invention, the compounds
according to the invention are administered in combination with a
thyroid receptor agonist, for example and preferably D-thyroxin,
3,5,3'-triiodothyronin (T3), CGS 23425 or axitirome (CGS
26214).
[0133] In a preferred embodiment of the invention, the compounds
according to the invention are administered in combination with an
HMG-CoA-reductase inhibitor from the class of statins, for example
and preferably lovastatin, simvastatin, pravastatin, fluvastatin,
atorvastatin, rosuvastatin or pitavastatin.
[0134] In a preferred embodiment of the invention, the compounds
according to the invention are administered in combination with a
squalene synthesis inhibitor, for example and preferably BMS-188494
or TAK-475.
[0135] In a preferred embodiment of the invention, the compounds
according to the invention are administered in combination with an
ACAT inhibitor, for example and preferably avasimibe, melinamide,
pactimibe, eflucimibe or SMP-797.
[0136] In a preferred embodiment of the invention, the compounds
according to the invention are administered in combination with an
MTP inhibitor, for example and preferably implitapide, R-103757,
BMS-201038 or JTT-130.
[0137] In a preferred embodiment of the invention, the compounds
according to the invention are administered in combination with a
PPAR-gamma agonist, for example and preferably pioglitazone or
rosiglitazone.
[0138] In a preferred embodiment of the invention, the compounds
according to the invention are administered in combination with a
PPAR-delta agonist, for example and preferably GW 501516 or BAY
68-5042.
[0139] In a preferred embodiment of the invention, the compounds
according to the invention are administered in combination with a
cholesterol absorption inhibitor, for example and preferably
ezetimibe, tiqueside or pamaqueside.
[0140] In a preferred embodiment of the invention, the compounds
according to the invention are administered in combination with a
lipase inhibitor, for example and preferably orlistat.
[0141] In a preferred embodiment of the invention, the compounds
according to the invention are administered in combination with a
polymeric bile acid adsorber, for example and preferably
cholestyramine, colestipol, colesolvam, CholestaGel or
colestimide.
[0142] In a preferred embodiment of the invention, the compounds
according to the invention are administered in combination with a
bile acid reabsorption inhibitor, for example and preferably ASBT
(=IBAT) inhibitors such as AZD-7806, S-8921, AK-105, BARI-1741,
SC-435 or SC-635.
[0143] In a preferred embodiment of the invention, the compounds
according to the invention are administered in combination with a
lipoprotein(a) antagonist, for example and preferably gemcabene
calcium (CI-1027) or nicotinic acid.
[0144] In a particularly preferred embodiment, the compounds of the
present invention are administered in combination with one or more
additional therapeutic agents selected from the group consisting of
diuretics, angiotensin AII antagonists, ACE inhibitors,
beta-receptor blockers, mineralocorticoid receptor antagonists,
organic nitrates, NO donors, activators of the soluble guanylate
cyclase (sGC), stimulators of the soluble guanylate cyclase and
positive-inotropic agents.
[0145] Thus, in a further embodiment, the present invention relates
to pharmaceutical compositions comprising at least one of the
compounds according to the invention and one or more additional
therapeutic agents for the treatment and/or prevention of diseases,
especially of the aforementioned diseases.
[0146] Furthermore, the compounds of the present invention may be
utilized, as such or in compositions, in research and diagnostics,
or as analytical reference standards and the like, which are well
known in the art.
[0147] When the compounds of the present invention are administered
as pharmaceuticals, to humans and other mammals, they can be given
per se or as a pharmaceutical composition containing, for example,
0.1% to 99.5% (more preferably, 0.5% to 90%) of active ingredient
in combination with one or more pharmaceutically acceptable
excipients.
[0148] Thus, in another aspect, the present invention relates to
pharmaceutical compositions comprising at least one of the
compounds according to the invention, conventionally together with
one or more inert, non-toxic, pharmaceutically suitable excipients,
and to the use thereof for the treatment and/or prevention of
diseases, especially of the aforementioned diseases.
[0149] The compounds according to the invention can act
systemically and/or locally. For this purpose, they can be
administered in a suitable way such as, for example, by the oral,
parenteral, pulmonary, nasal, lingual, sublingual, buccal, rectal,
dermal, transdermal, conjunctival, otic or topical route, or as an
implant or stent.
[0150] For these administration routes, the compounds of the
invention can be administered in suitable application forms.
[0151] Suitable for oral administration are application forms which
function according to the state of the art and deliver the
compounds according to the invention rapidly and/or in modified
fashion, and which contain the compounds according to the invention
in crystalline, amorphous and/or dissolved form, such as, for
example, tablets (uncoated or coated tablets, for example having
enteric coatings or coatings which are insoluble or dissolve with a
delay and control the release of the compound according to the
invention), tablets which disintegrate rapidly in the mouth, or
films/wafers, films/lyophilisates, capsules (e.g. hard or soft
gelatin capsules), sugar-coated tablets, granules, pellets,
powders, emulsions, suspensions, aerosols or solutions.
[0152] Parenteral application can be carried out with avoidance of
an absorption step (intravenously, intraarterially, intracardially,
intraspinally or intralumbarly) or with inclusion of an absorption
(intramuscularly, subcutaneously, intracutaneously, percutaneously
or intraperitoneally). Suitable parenteral application forms
include injection and infusion preparations in the form of
solutions, suspensions, emulsions, lyophilisates and sterile
powders.
[0153] Forms suitable for other application routes include, for
example, inhalatory pharmaceutical forms (e.g. powder inhalers,
nebulizers), nasal drops, solutions or sprays, tablets or capsules
to be administered lingually, sublingually or buccally (e.g.
troches, lozenges), suppositories, ear and eye preparations (e.g.
drops, ointments), vaginal capsules, aqueous suspensions (lotions,
shaking mixtures), lipophilic suspensions, ointments, creams,
milks, pastes, foams, dusting powders, transdermal therapeutic
systems (e.g. patches), implants and stents.
[0154] In a preferred embodiment, the pharmaceutical composition
comprising a compound of formula (I) as defined above is provided
in a form suitable for oral administration. In another preferred
embodiment, the pharmaceutical composition comprising a compound of
formula (I) as defined above is provided in a form suitable for
intravenous administration.
[0155] The compounds according to the invention can be converted
into the recited application forms in a manner known per se by
mixing with inert, non-toxic, pharmaceutically suitable excipients.
These excipients include, inter alia, carriers (e.g.
microcrystalline cellulose, lactose, mannitol), solvents (e.g.
liquid polyethylene glycols), emulsifiers (e.g. sodium dodecyl
sulfate), surfactants (e.g. polyoxysorbitan oleate), dispersants
(e.g. polyvinylpyrrolidone), synthetic and natural polymers (e.g.
albumin), stabilizers (e.g. antioxidants such as, for example,
ascorbic acid), colorants (e.g. inorganic pigments such as, for
example, iron oxides), and flavour and/or odour masking agents.
[0156] A preferred dose of the compound of the present invention is
the maximum that a patient can tolerate and not develop serious
side effects. Illustratively, the compound of the present invention
may be administered parenterally at a dose of about 0.001 mg/kg to
about 10 mg/kg, preferably of about 0.01 mg/kg to about 1 mg/kg of
body weight. In oral administration, an exemplary dose range is
about 0.01 to 100 mg/kg, preferably about 0.01 to 20 mg/kg, and
more preferably about 0.1 to 10 mg/kg of body weight. Ranges
intermediate to the above-recited values are also intended to be
part of the invention.
[0157] Nevertheless, actual dosage levels and time course of
administration of the active ingredients in the pharmaceutical
compositions of the invention may be varied so as to obtain an
amount of the active ingredient which is effective to achieve the
desired therapeutic response for a particular patient, composition
and mode of administration, without being toxic to the patient. It
may therefore be necessary where appropriate to deviate from the
stated amounts, in particular as a function of age, gender, body
weight, diet and general health status of the patient, the
bioavailability and pharmacodynamic characteristics of the
particular compound and its mode and route of administration, the
time or interval over which administration takes place, the dose
regimen selected, the response of the individual patient to the
active ingredient, the specific disease involved, the degree of or
the involvement or severity of the disease, the kind of concurrent
treatment (i.e., the interaction of the compound of the invention
with other co-administered therapeutics), and other relevant
circumstances.
[0158] Thus, it may be satisfactory in some cases to manage with
less than the aforementioned minimum amount, whereas in other cases
the stated upper limit must be exceeded. Treatment can be initiated
with smaller dosages, which are less than the optimum dose of the
compound. Thereafter, the dosage may be increased by small
increments until the optimum effect under the circumstances is
reached. For convenience, the total daily dosage may be divided and
administered in individual portions spread over the day.
[0159] The following exemplary embodiments illustrate the
invention. The invention is not restricted to the examples.
[0160] The percentages in the following tests and examples are,
unless stated otherwise, by weight; parts are by weight. Solvent
ratios, dilution ratios and concentrations reported for
liquid/liquid solutions are each based on volume.
A. EXAMPLES
Abbreviations and Acronyms
[0161] Ac acetyl [0162] aq. aqueous (solution) [0163] br. broad
(.sup.1H NMR signal) [0164] cat. catalytic [0165] conc.
concentrated [0166] d doublet (.sup.1H NMR signal) [0167] DCI
direct chemical ionization (MS) [0168] d.e. diastereomeric excess
[0169] DMF N,N-dimethylformamide [0170] DMSO dimethylsulfoxide
[0171] EI electron impact ionization (MS) [0172] eq. equivalent(s)
[0173] ESI electro-spray ionization (MS) [0174] Et ethyl [0175] h
hour(s) [0176] .sup.1H NMR proton nuclear magnetic resonance
spectroscopy [0177] HPLC high performance liquid chromatography
[0178] LC/MS liquid chromatography-coupled mass spectroscopy [0179]
m multiplet (.sup.1H NMR signal) [0180] Me methyl [0181] min
minute(s) [0182] MS mass spectroscopy [0183] MTBE methyl tert-butyl
ether [0184] m/z mass-to-charge ratio (MS) [0185] of th. of theory
(chemical yield) [0186] q quartet (.sup.1H NMR signal) [0187]
quant. quantitative (yield) [0188] rac racemic [0189] R.sub.f TLC
retention factor [0190] RP reverse phase (HPLC) [0191] rt room
temperature [0192] R.sub.t retention time (HPLC) [0193] s singlet
(.sup.1H NMR signal) [0194] sat. saturated (solution) [0195] SFC
supercritical fluid chromatography [0196] t triplet (.sup.1H NMR
signal) [0197] tBu tert-butyl [0198] tert tertiary [0199] TFA
trifluoroacetic acid [0200] THF tetrahydrofuran [0201] TLC thin
layer chromatography [0202] UV ultraviolet
LC/MS and HPLC Methods:
Method 1 (LC/MS):
[0203] Instrument: Waters Acquity SQD UPLC System; column: Waters
Acquity UPLC HSS T3 1.8.mu., 50 mm.times.1 mm; eluent A: 1 L
water+0.25 mL 99% formic acid, eluent B: 1 L acetonitrile+0.25 mL
99% formic acid; gradient: 0.0 min 90% A.fwdarw.1.2 min 5%
A.fwdarw.2.0 min 5% A; oven: 50.degree. C.; flow rate: 0.40 mL/min;
UV detection: 208-400 nm.
Method 2 (LC/MS):
[0204] Instrument: Waters Acquity SQD UPLC System; column: Waters
Acquity UPLC HSS T3 1.8.mu., 50 mm.times.1 mm; eluent A: 1 L
water+0.25 mL 99% formic acid, eluent B: 1 L acetonitrile+0.25 mL
99% formic acid; gradient: 0.0 min 95% A.fwdarw.6.0 min 5%
A.fwdarw.7.5 min 5% A; oven: 50.degree. C.; flow rate: 0.35 mL/min;
UV detection: 210-400 nm.
Method 3 (LC/MS):
[0205] Instrument MS: Agilent MS Quad 6150; Instrument HPLC:
Agilent 1290; column: Waters Acquity UPLC HSS T3 1.8.mu., 50
mm.times.2.1 mm; eluent A: 1 L water+0.25 mL 99% formic acid,
eluent B: 1 L acetonitrile+0.25 mL 99% formic acid; gradient: 0.0
min 90% A.fwdarw.0.3 min 90% A.fwdarw.1.7 min 5% A.fwdarw.3.0 min
5% A; oven: 50.degree. C.; flow rate: 1.20 mL/min; UV detection:
205-305 nm.
Method 4 (Preparative HPLC):
[0206] Column: Chromatorex C18 10 .mu.m, 125 mm.times.30 mm; eluent
A: water+0.05% TFA, eluent B: acetonitrile+0.05% TFA; gradient: 20%
B.fwdarw.45% B, 45% B isocratic, 45% B.fwdarw.80% B; column
temperature: room temperature; flow rate: 50 mL/min; UV detection:
210 nm.
Starting Materials and Intermediates:
Example 1A
Methyl
{3-(4-chlorophenyl)-5-oxo-4-[(2S)-3,3,3-trifluoro-2-hydroxypropyl]--
4,5-dihydro-1H-1,2,4-triazol-1-yl}acetate
##STR00013##
[0208] Under argon, potassium tert-butoxide (9.118 g, 81.26 mmol)
was added portionwise at room temperature to a solution of
5-(4-chlorophenyl)-4-[(2S)-3,3,3-trifluoro-2-hydroxypropyl]-2,4-dihydro-3-
H-1,2,4-triazol-3-one (Example 5A in WO 2011/104322-A1; 20 g, 65.01
mmol) in THF (40 ml). To this solution was added methyl
bromoacetate (10.939 g, 71.51 mmol), and the mixture was stirred at
room temperature overnight. The reaction mixture was then diluted
with water and extracted with ethyl acetate. The combined organic
phases were dried over sodium sulfate, filtered, and concentrated
in vacuo. 16.4 g (30.23 mmol) of the desired compound were obtained
(46.5% yield, 70% purity).
[0209] LC/MS [method 1]: R.sub.t=0.90 min; MS [ESIpos]: m/z=380
(M+H).sup.+
[0210] .sup.1H NMR (400 MHz, DMSO-d.sub.6): .delta. [ppm] 3.70 (s,
3H), 3.85 (dd, 1H), 4.00 (dd, 1H), 4.19-4.33 (m, 1H), 4.72 (s, 2H),
6.92 (d, 1H), 7.60-7.69 (m, 2H), 7.73-7.81 (m, 2H).
[0211] The title compound can also be synthesized via the procedure
described in WO 2011/104322-A1 (Example 7A).
Example 2A
2-{3-(4-Chlorophenyl)-5-oxo-4-[(2S)-3,3,3-trifluoro-2-hydroxypropyl]-4,5-d-
ihydro-1H-1,2,4-triazol-1-yl}acetohydrazide
##STR00014##
[0213] 7.2 g (18.96 mmol) of methyl
{3-(4-chlorophenyl)-5-oxo-4-[(2S)-3,3,3-trifluoro-2-hydroxypropyl]-4,5-di-
hydro-1H-1,2,4-triazol-1-yl}acetate were dissolved in 60 ml of
absolute ethanol. To this solution were added 2.088 g (41.71 mmol)
of hydrazine hydrate, and the mixture was stirred under reflux for
5 h and then at room temperature overnight. The resulting mixture
was partially concentrated in vacuo and then diluted with water and
extracted with ethyl acetate. The combined organic phases were
dried over sodium sulfate, filtered, and concentrated in vacuo. The
residue was dissolved in dichloromethane, and after crystallization
the white solid was filtered off and dried under high vacuum. 7.02
g (18.49 mmol) of the desired compound were obtained (97.5%
yield).
[0214] LC/MS [method 1]: R.sub.t=0.73 min; MS [ESIpos]: m/z=380
(M+H).sup.+
[0215] .sup.1H NMR (400 MHz, DMSO-d.sub.6): .delta. [ppm] 3.82 (dd,
1H), 3.96 (dd, 1H), 4.24-4.34 (m, 3H), 4.38 (d, 2H), 6.90 (d, 1H),
7.61-7.66 (m, 2H), 7.73-7.78 (m, 2H), 9.23 (t, 1H).
Example 3A
5-(4-Chlorophenyl)-2-{[5-(hydroxymethyl)-1H-1,2,4-triazol-3-yl]methyl}-4-[-
(2S)-3,3,3-trifluoro-2-hydroxypropyl]-2,4-dihydro-3H-1,2,4-triazol-3-one
##STR00015##
[0217] Under argon, sodium ethoxide (2.987 g, 42.14 mmol, 96%
purity) was added portionwise at room temperature to a solution of
2-{3-(4-chlorophenyl)-5-oxo-4-[(2S)-3,3,3-trifluoro-2-hydroxypropyl]-4,5--
dihydro-1H-1,2,4-triazol-1-yl}acetohydrazide (8.0 g, 21.07 mmol)
and 2-hydroxyacetamidine hydrochloride (2.329 g, 21.07 mmol) in DMF
(200 ml). The reaction mixture was stirred at 100.degree. C.
overnight. After cooling, the reaction mixture was partially
concentrated in vacuo and then diluted with ethyl acetate. The
resulting mixture was washed with water, and after phase
separation, the aqueous phase was extracted twice with ethyl
acetate. The combined organic phases were dried over sodium
sulfate, filtered, and concentrated under reduced pressure. The
resulting solid was dried under high vacuum to give 8.69 g (89%
purity, 18.47 mmol) of the desired compound which was used without
further purification (.about.88% yield).
[0218] LC/MS [method 1]: R.sub.t=0.74 min; MS [ESIpos]: m/z=419
(M+H).sup.+
[0219] .sup.1H NMR (400 MHz, DMSO-d.sub.6): .delta. [ppm] 3.83 (dd,
1H), 3.98 (dd, 1H), 4.24-4.36 (m, 1H), 4.53 (br. s, 2H), 4.96 (br.
s, 2H), 5.64 (br. s, 1H), 6.91 (d, 1H), 7.58-7.67 (m, 2H),
7.72-7.78 (m, 2H), 13.75 (br. s, 1H).
Example 4A
5-(4-Chlorophenyl)-2-({5-[(1RS)-1-hydroxyethyl]-1H-1,2,4-triazol-3-yl}meth-
yl)-4-[(2S)-3,3,3-trifluoro-2-hydroxypropyl]-2,4-dihydro-3H-1,2,4-triazol--
3-one (diastereomeric mixture)
##STR00016##
[0221] Under argon, sodium ethoxide (1.531 g, 21.59 mmol, 96%
purity) was added portionwise at room temperature to a solution of
2-{3-(4-chlorophenyl)-5-oxo-4-[(2S)-3,3,3-trifluoro-2-hydroxypropyl]-4,5--
dihydro-1H-1,2,4-triazol-1-yl}acetohydrazide (4.1 g, 10.80 mmol)
and 2-hydroxypropanimidamide hydrochloride (1.480 g, 11.88 mmol) in
DMF (110 ml). The reaction mixture was stirred at 120.degree. C.
for 4.5 h. After cooling, the reaction mixture was partially
concentrated in vacuo and then diluted with ethyl acetate. The
resulting mixture was washed with water, and after phase
separation, the aqueous phase was extracted twice with ethyl
acetate. The combined organic phases were dried over sodium
sulfate, filtered, and concentrated under reduced pressure. The
resulting solid was dried under high vacuum to give 4.90 g (92%
purity, 10.42 mmol) of the desired compound as a mixture of
diastereomers which was used without further purification.
[0222] LC/MS [method 1]: R.sub.t=0.82 min; MS [ESIpos]: m/z=433
(M+H).sup.+
[0223] .sup.1H NMR (400 MHz, DMSO-d.sub.6): .delta. [ppm] 1.39 (d,
3H), 3.79-3.88 (m, 1H), 3.93-4.02 (m, 1H), 4.24-4.36 (m, 1H), 4.80
(quin, 1H), 4.89-5.00 (m, 2H), 5.73 (d, 1H), 6.93 (d, 1H),
7.58-7.65 (m, 2H), 7.70-7.77 (d, 2H), 13.68 (s, 1H).
Example 5A
5-(4-Chlorophenyl)-2-({5-[(1
S)-1-hydroxyethyl]-1H-1,2,4-triazol-3-yl}methyl)-4-[(2S)-3,3,3-trifluoro--
2-hydroxypropyl]-2,4-dihydro-3H-1,2,4-triazol-3-one
##STR00017##
[0225] The title compound was synthesized analogously to Example 4A
starting from (2S)-2-hydroxypropanimidamide hydrochloride (1.1
eq.), using only 1.1 eq. of sodium ethoxide as base (reaction
temperature: 100.degree. C.).
[0226] LC/MS [method 1]: R.sub.t=0.81 min; MS [ESIpos]: m/z=433
(M+H).sup.+.
[0227] (2S)-2-hydroxypropanimidamide hydrochloride, which is
commercially available, was also synthesized from
(2S)-2-hydroxypropanamide [L-(-)-lactamide] via the procedure
described in WO 00/59510-A1 (Preparation 11, Step A) and WO
2013/138860-A1 (Intermediate to Precursor 82, p. 101-102).
Example 6A
5-(4-Chlorophenyl)-2-({5-[(1R)-1-hydroxyethyl]-1H-1,2,4-triazol-3-yl}methy-
l)-4-[(2S)-3,3,3-trifluoro-2-hydroxypropyl]-2,4-dihydro-3H-1,2,4-triazol-3-
-one
##STR00018##
[0229] The title compound was synthesized analogously to Example 4A
starting from (2R)-2-hydroxypropanimidamide hydrochloride (1.1
eq.), using only 1.1 eq. of sodium ethoxide as base (reaction
temperature: 100.degree. C.).
[0230] LC/MS [method 3]: R.sub.t=1.00 min; MS [ESIpos]: m/z=433
(M+H).sup.+.
[0231] (2R)-2-hydroxypropanimidamide hydrochloride, which is
commercially available, was also synthesized from
(2R)-2-hydroxypropanamide [R-(+)-lactamide] via the procedure
described in WO 00/59510-A1 (Preparation 11, Step A) and WO
2013/138860-A1 (Intermediate to Precursor 82, p. 101-102).
Preparation Examples
[0232] Example compounds bearing a 1-hydroxyethyl substituent
[R.sup.1 in formula (I)=CH.sub.3] which in the following are termed
"diastereomer 1" and "diastereomer 2", respectively, represent
pairs of separated diastereomers whose absolute configuration with
regard to the 1-hydroxyethyl moiety (1R or 1S) had not been
determined.
[0233] Diastereomeric excess (d.e.) values were determined in the
usual way by analysis of HPLC peak areas according to the following
formula:
d . e . = diastereomer 1 ( % peak area ) - diastereomer 2 ( % peak
area ) diastereomer 1 ( % peak area ) + diastereomer 2 ( % peak
area ) .times. 100 % . ##EQU00001##
Example 1
5-(4-Chlorophenyl)-2-{[1-(3-chlorophenyl)-5-(hydroxymethyl)-1H-1,2,4-triaz-
ol-3-yl]methyl}-4-[(2S)-3,3,3-trifluoro-2-hydroxypropyl]-2,4-dihydro-3H-1,-
2,4-triazol-3-one
##STR00019##
[0235] To a solution of
5-(4-chlorophenyl)-2-{[5-(hydroxymethyl)-1H-1,2,4-triazol-3-yl]methyl}-4--
[(2S)-3,3,3-trifluoro-2-hydroxypropyl]-2,4-dihydro-3H-1,2,4-triazol-3-one
(400 mg, 0.96 mmol) in pyridine (12 ml) were added
(3-chlorophenyl)boronic acid (298.74 mg, 1.91 mmol) and copper(II)
acetate (347 mg, 1.91 mmol). The reaction mixture was stirred at
room temperature for 5 days, after which extra boronic acid (74.7
mg, 0.48 mmol, 0.5 eq.) was added due to incomplete conversion.
After stirring for two additional days, the reaction mixture was
diluted with MTBE and then quenched with aqueous hydrochloric acid
(0.5 M). After phase separation, the aqueous phase was extracted
twice with MTBE. The combined organic phases were dried over sodium
sulfate, filtered, and concentrated in vacuo. The crude product was
purified by preparative HPLC [method 4], and the desired compound
(113 mg, 0.21 mmol) was obtained (yield 22.4%).
[0236] LC/MS [method 2]: R.sub.t=3.07 min; MS [ESIpos]: m/z=529
(M+H).sup.+
[0237] .sup.1H NMR (400 MHz, DMSO-d.sub.6): .delta. [ppm] 3.85 (dd,
1H), 4.01 (dd, 1H), 4.30 (br. s, 1H), 4.58 (s, 2H), 5.08 (s, 2H),
6.90 (br. d, 1H), 7.55-7.64 (m, 4H), 7.65-7.69 (m, 1H), 7.73-7.78
(m, 2H), 7.78-7.81 (m, 1H).
Example 2
5-(4-Chlorophenyl)-2-{[1-(3-fluorophenyl)-5-(hydroxymethyl)-1H-1,2,4-triaz-
ol-3-yl]methyl}-4-[(2S)-3,3,3-trifluoro-2-hydroxypropyl]-2,4-dihydro-3H-1,-
2,4-triazol-3-one
##STR00020##
[0239] To a solution of
5-(4-chlorophenyl)-2-{[5-(hydroxymethyl)-1H-1,2,4-triazol-3-yl]methyl}-4--
[(2S)-3,3,3-trifluoro-2-hydroxypropyl]-2,4-dihydro-3H-1,2,4-triazol-3-one
(100 mg, 0.24 mmol) in pyridine (3 ml) were added
(3-fluorophenyl)boronic acid (66.83 mg, 0.48 mmol) and copper(II)
acetate (86.75 mg, 0.48 mmol). The reaction mixture was stirred at
room temperature for 5 days, after which extra boronic acid (16.72
mg, 0.12 mmol, 0.5 eq.) was added due to incomplete conversion.
After stirring for two additional days, the reaction mixture was
diluted with MTBE and then quenched with aqueous hydrochloric acid
(0.5 M). After phase separation, the aqueous phase was extracted
twice with MTBE. The combined organic phases were dried over sodium
sulfate, filtered, and concentrated in vacuo. The crude product was
purified by preparative HPLC [method 4], and the desired compound
(25 mg, 0.05 mmol) was obtained (yield 20.2%).
[0240] LC/MS [method 2]: R.sub.t=2.87 min; MS [ESIpos]: m/z=513
(M+H).sup.+
[0241] .sup.1H NMR (400 MHz, DMSO-d.sub.6): .delta. [ppm] 3.85 (dd,
1H), 4.01 (dd, 1H), 4.30 (br. s, 1H), 4.59 (s, 2H), 5.08 (s, 2H),
6.90 (br. d, 1H), 7.37 (td, 1H), 7.51-7.66 (m, 5H), 7.72-7.79 (m,
2H).
Example 3
5-(4-Chlorophenyl)-2-{[5-(hydroxymethyl)-1-(2-methoxyphenyl)-1H-1,2,4-tria-
zol-3-yl]methyl}-4-[(2S)-3,3,3-trifluoro-2-hydroxypropyl]-2,4-dihydro-3H-1-
,2,4-triazol-3-one
##STR00021##
[0243] To a solution of
5-(4-chlorophenyl)-2-{[5-(hydroxymethyl)-1H-1,2,4-triazol-3-yl]methyl}-4--
[(2S)-3,3,3-trifluoro-2-hydroxypropyl]-2,4-dihydro-3H-1,2,4-triazol-3-one
(300 mg, 0.72 mmol) in pyridine (9 ml) were added
(2-methoxyphenyl)boronic acid (217.72 mg, 1.43 mmol) and copper(II)
acetate (260.25 mg, 1.43 mmol). The reaction mixture was stirred at
room temperature for 5 days, after which extra boronic acid (54.4
mg, 0.36 mmol, 0.5 eq.) was added due to incomplete conversion.
After stirring for two additional days, the reaction mixture was
diluted with MTBE and then quenched with aqueous hydrochloric acid
(0.5 M). After phase separation, the aqueous phase was extracted
twice with MTBE. The combined organic phases were dried over sodium
sulfate, filtered, and concentrated in vacuo. The crude product was
purified by preparative HPLC [method 4], and the desired compound
(73 mg, 0.14 mmol) was obtained (yield 22.4%, 98% purity).
[0244] LC/MS [method 3]: R.sub.t=1.18 min; MS [ESIpos]: m/z=525
(M+H).sup.+
[0245] .sup.1H NMR (400 MHz, DMSO-d.sub.6): .delta. [ppm] 3.77 (s,
3H), 3.85 (dd, 1H), 4.01 (dd, 1H), 4.31 (br. s, 1H), 4.35 (s, 2H),
5.04 (s, 2H), 6.91 (br. s, 1H), 7.09 (t, 1H), 7.25 (d, 1H), 7.37
(dd, 1H), 7.52 (td, 1H), 7.63 (d, 2H), 7.76 (d, 2H).
Example 4
5-(4-Chlorophenyl)-2-{[1-(2-chlorophenyl)-5-(hydroxymethyl)-1H-1,2,4-triaz-
ol-3-yl]methyl}-4-[(2S)-3,3,3-trifluoro-2-hydroxypropyl]-2,4-dihydro-3H-1,-
2,4-triazol-3-one
##STR00022##
[0247] To a solution of
5-(4-chlorophenyl)-2-{[5-(hydroxymethyl)-1H-1,2,4-triazol-3-yl]methyl}-4--
[(2S)-3,3,3-trifluoro-2-hydroxypropyl]-2,4-dihydro-3H-1,2,4-triazol-3-one
(3 g, 5.946 mmol, 83% purity) in pyridine (75 ml) were added
(2-chlorophenyl)boronic acid (930 mg, 5.946 mmol) and copper(II)
acetate (2.16 g, 11.89 mmol). The reaction mixture was stirred at
room temperature overnight, after which extra boronic acid (500 mg,
3.20 mmol) was added due to incomplete conversion. The reaction
mixture was further stirred at room temperature for 9 days. Over
this time, three additional portions of boronic acid (1.5 g in
total, 9.6 mmol) were added. After this, the reaction mixture was
diluted with MTBE and then quenched with aqueous hydrochloric acid
(0.5 M). After phase separation, the aqueous phase was extracted
twice with MTBE. The combined organic phases were dried over sodium
sulfate, filtered, and concentrated in vacuo. The crude product was
purified by preparative HPLC [method 4], and the desired compound
(1.44 g, 2.72 mmol) was obtained (yield 45.7%).
[0248] LC/MS [method 2]: R.sub.t=2.79 min; MS [ESIpos]: m/z=529
(M+H).sup.+
[0249] .sup.1H NMR (400 MHz, DMSO-d.sub.6): .delta. [ppm] 3.85 (dd,
1H), 4.01 (dd, 1H), 4.31 (br. s, 1H), 4.41 (s, 2H), 5.07 (s, 2H),
6.91 (d, 1H), 7.49-7.67 (m, 5H), 7.68-7.80 (m, 3H).
Example 5
5-(4-Chlorophenyl)-2-{[5-(hydroxymethyl)-1-(2-methylphenyl)-1H-1,2,4-triaz-
ol-3-yl]methyl}-4-[(2S)-3,3,3-trifluoro-2-hydroxypropyl]-2,4-dihydro-3H-1,-
2,4-triazol-3-one
##STR00023##
[0251] To a solution of
5-(4-chlorophenyl)-2-{[5-(hydroxymethyl)-1H-1,2,4-triazol-3-yl]methyl}-4--
[(2S)-3,3,3-trifluoro-2-hydroxypropyl]-2,4-dihydro-3H-1,2,4-triazol-3-one
(400 mg, 0.96 mmol) in pyridine (12 ml) were added
(2-methylphenyl)boronic acid (259.7 mg, 1.91 mmol) and copper(II)
acetate (347 mg, 1.91 mmol). The reaction mixture was stirred at
room temperature for 5 days, after which extra boronic acid (64.9
mg, 0.48 mmol, 0.5 eq.) was added due to incomplete conversion.
After stirring for two additional days, the reaction mixture was
diluted with MTBE and then quenched with aqueous hydrochloric acid
(0.5 M). After phase separation, the aqueous phase was extracted
twice with MTBE. The combined organic phases were dried over sodium
sulfate, filtered, and concentrated in vacuo. The crude product was
purified by preparative HPLC [method 4], and the desired compound
(58 mg, 0.11 mmol) was obtained (yield 11.9%).
[0252] LC/MS [method 2]: R.sub.t=2.85 min; MS [ESIpos]: m/z=509
(M+H).sup.+
[0253] .sup.1H NMR (400 MHz, DMSO-d.sub.6): .delta. [ppm] 2.01 (s,
3H), 3.85 (dd, 1H), 4.00 (dd, 1H), 4.30 (br. s, 1H), 4.34 (s, 2H),
5.07 (s, 2H), 6.90 (br. s, 1H), 7.32-7.50 (m, 4H), 7.62 (br. d,
2H), 7.75 (br. d, 2H).
Example 6
2-{[1-(3-Chloro-5-fluorophenyl)-5-(hydroxymethyl)-1H-1,2,4-triazol-3-yl]me-
thyl}-5-(4-chlorophenyl)-4-[(2S)-3,3,3-trifluoro-2-hydroxypropyl]-2,4-dihy-
dro-3H-1,2,4-triazol-3-one
##STR00024##
[0255] To a solution of
5-(4-chlorophenyl)-2-{[5-(hydroxymethyl)-1H-1,2,4-triazol-3-yl]methyl}-4--
[(2S)-3,3,3-trifluoro-2-hydroxypropyl]-2,4-dihydro-3H-1,2,4-triazol-3-one
(400 mg, 0.96 mmol) in pyridine (12 ml) were added
(3-chloro-5-fluorophenyl)boronic acid (333.1 mg, 1.91 mmol) and
copper(II) acetate (347 mg, 1.91 mmol). The reaction mixture was
stirred at room temperature for 5 days, after which extra boronic
acid (83.2 mg, 0.48 mmol, 0.5 eq.) was added due to incomplete
conversion. After stirring for two additional days, the reaction
mixture was diluted with MTBE and then quenched with aqueous
hydrochloric acid (0.5 M). After phase separation, the aqueous
phase was extracted twice with MTBE. The combined organic phases
were dried over sodium sulfate, filtered, and concentrated in
vacuo. The crude product was purified by preparative HPLC [method
4], and the desired compound (91 mg, 0.17 mmol) was obtained (yield
17.4%).
[0256] LC/MS [method 3]: R.sub.t=1.31 min; MS [ESIpos]: m/z=547
(M+H).sup.+
[0257] .sup.1H NMR (400 MHz, DMSO-d.sub.6): .delta. [ppm] 3.85 (dd,
1H), 4.01 (dd, 1H), 4.30 (br. s, 1H), 4.63 (s, 2H), 5.08 (s, 2H),
6.90 (br. s, 1H), 7.59-7.66 (m, 4H), 7.67-7.71 (m, 1H), 7.72-7.78
(m, 2H).
Example 7
5-(4-Chlorophenyl)-2-{[1-(3,5-difluorophenyl)-5-(hydroxymethyl)-1H-1,2,4-t-
riazol-3-yl]methyl}-4-[(2S)-3,3,3-trifluoro-2-hydroxypropyl]-2,4-dihydro-3-
H-1,2,4-triazol-3-one
##STR00025##
[0259] The title compound was prepared analogously to Example 1
starting from 360 mg (0.86 mmol) of
5-(4-chlorophenyl)-2-{[5-(hydroxymethyl)-1H-1,2,4-triazol-3-yl]methyl}-4--
[(2S)-3,3,3-trifluoro-2-hydroxypropyl]-2,4-dihydro-3H-1,2,4-triazol-3-one.
61 mg (0.11 mmol) of the desired compound were obtained (13.4%
yield).
[0260] LC/MS [method 3]: R.sub.t=1.25 min; MS [ESIpos]: m/z=531
(M+H).sup.+
[0261] .sup.1H NMR (500 MHz, DMSO-d.sub.6): .delta. [ppm] 3.86 (dd,
1H), 4.01 (dd, 1H), 4.25-4.38 (m, 1H), 4.64 (d, 2H), 5.08 (s, 2H),
5.94 (t, 1H), 6.92 (d, 1H), 7.46 (tt, 1H), 7.49-7.55 (m, 2H), 7.62
(br. d, 2H), 7.76 (br. d, 2H).
Example 8
5-(4-Chlorophenyl)-2-({5-(hydroxymethyl)-1-[2-(trifluoromethyl)phenyl]-H-1-
,2,4-triazol-3-yl}-methyl)-4-[(2S)-3,3,3-trifluoro-2-hydroxypropyl]-2,4-di-
hydro-3H-1,2,4-triazol-3-one
##STR00026##
[0263] The title compound was prepared analogously to Example 1
starting from 500 mg (1.19 mmol) of
5-(4-chlorophenyl)-2-{[5-(hydroxymethyl)-1H-1,2,4-triazol-3-yl]methyl}-4--
[(2S)-3,3,3-trifluoro-2-hydroxypropyl]-2,4-dihydro-3H-1,2,4-triazol-3-one.
104 mg (0.18 mmol) of the desired compound were obtained (15.5%
yield).
[0264] LC/MS [method 3]: R.sub.t=1.24 min; MS [ESIpos]: m/z=563
(M+H).sup.+
[0265] .sup.1H NMR (400 MHz, DMSO-d.sub.6): .delta. [ppm] 3.85 (dd,
1H), 4.00 (dd, 1H), 4.30 (br. s, 1H), 4.39 (s, 2H), 5.01-5.12 (m,
2H), 5.53 (br. s, 1H), 6.92 (d, 1H), 7.63 (d, 2H), 7.70 (d, 1H),
7.75 (d, 2H), 7.78-7.91 (m, 2H), 7.97 (d, 1H).
Example 9
2-{[1-(2-Chloro-5-fluorophenyl)-5-(hydroxymethyl)-1H-1,2,4-triazol-3-yl]me-
thyl}-5-(4-chlorophenyl)-4-[(2S)-3,3,3-trifluoro-2-hydroxypropyl]-2,4-dihy-
dro-3H-1,2,4-triazol-3-one
##STR00027##
[0267] The title compound was prepared analogously to Example 1
starting from 500 mg (1.19 mmol) of
5-(4-chlorophenyl)-2-{[5-(hydroxymethyl)-1H-1,2,4-triazol-3-yl]methyl}-4--
[(2S)-3,3,3-trifluoro-2-hydroxypropyl]-2,4-dihydro-3H-1,2,4-triazol-3-one.
8.7 mg (0.02 mmol) of the desired compound were obtained (1.3%
yield, 95% purity).
[0268] LC/MS [method 3]: R.sub.t=1.23 min; MS [ESIpos]: m/z=547
(M+H).sup.+
[0269] .sup.1H NMR (500 MHz, DMSO-d.sub.6): .delta. [ppm] 3.86 (dd,
1H), 4.01 (dd, 1H), 4.30 (br. s, 1H), 4.46 (s, 2H), 5.03-5.11 (m,
2H), 6.92 (br. d, 1H), 7.53 (td, 1H), 7.61-7.65 (m, 2H), 7.68 (dd,
1H), 7.73-7.80 (m, 3H).
Example 10
5-(4-Chlorophenyl)-2-({5-[(1RS)-1-hydroxyethyl]-1-(2-methoxyphenyl)-1H-1,2-
,4-triazol-3-yl}-methyl)-4-[(2S)-3,3,3-trifluoro-2-hydroxypropyl]-2,4-dihy-
dro-3H-1,2,4-triazol-3-one (diastereomeric mixture)
##STR00028##
[0271] To a solution of
5-(4-chlorophenyl)-2-({5-[(1RS)-1-hydroxyethyl]-1H-1,2,4-triazol-3-yl}met-
hyl)-4-[(2S)-3,3,3-trifluoro-2-hydroxypropyl]-2,4-dihydro-3H-1,2,4-triazol-
-3-one (500 mg, 1.16 mmol) in pyridine (15 ml) were added
(2-methoxyphenyl)boronic acid (351.1 mg, 2.31 mmol) and copper(II)
acetate (419.7 mg, 2.31 mmol). The reaction mixture was stirred at
room temperature for 5 days, after which extra boronic acid (87 mg,
0.58 mmol, 0.5 eq.) was added due to incomplete conversion. After
stirring for two additional days, the reaction mixture was
concentrated in vacuo, then diluted with MTBE and quenched with
aqueous hydrochloric acid (0.5 M). After phase separation, the
aqueous phase was extracted twice with MTBE. The combined organic
phases were dried over sodium sulfate, filtered, and concentrated
in vacuo. The crude product was purified by preparative HPLC
[method 4], and the desired compound (132 mg, 0.22 mmol) was
obtained as a mixture of diastereomers (yield 19.1%, 90%
purity).
[0272] LC/MS [method 2]: R.sub.t=2.87 min; MS [ESIpos]: m/z=539
(M+H).sup.+
[0273] .sup.1H NMR (400 MHz, DMSO-d.sub.6): .delta. [ppm] 1.36 (d,
3H), 3.76 (s, 3H), 3.85 (dd, 1H), 4.00 (dd, 1H), 4.30 (br. s, 1H),
4.47-4.61 (m, 1H), 4.98-5.10 (m, 2H), 6.90 (br. s, 1H), 7.09 (td,
1H), 7.24 (dd, 1H), 7.35 (dd, 1H), 7.49-7.55 (m, 1H), 7.60-7.65 (m,
2H), 7.73-7.79 (m, 2H).
[0274] The two diastereomers were separated by preparative chiral
HPLC [sample preparation: 128 mg dissolved in 10 ml methanol;
injection volume: 0.3 ml; column: Daicel Chiralcel.RTM. OX-H 5
.mu.m, 250.times.20 mm; eluent: isohexane/methanol 70:30; flow
rate: 80 ml/min; temperature: 40.degree. C.; UV detection: 210 nm].
After separation, 43.6 mg of diastereomer 1 (Example 11), which
eluted first, and 45.1 mg of diastereomer 2 (Example 12), which
eluted later, were isolated.
Example 11
5-(4-Chlorophenyl)-2-{[5-(1-hydroxyethyl)-1-(2-methoxyphenyl)-1H-1,2,4-tri-
azol-3-yl]methyl}-4-[(2S)-3,3,3-trifluoro-2-hydroxypropyl]-2,4-dihydro-3H--
1,2,4-triazol-3-one (diastereomer 1)
[0275] Analytical chiral HPLC (SFC): R.sub.t=3.08 min, d.e.=100%
[column: Daicel Chiralcel.RTM. OX-3 250.times.4 mm; eluent: carbon
dioxide/methanol (5%.fwdarw.60%); flow rate: 3 ml/min; UV
detection: 220 nm].
[0276] .sup.1H NMR (400 MHz, DMSO-d.sub.6): .delta. [ppm] 1.36 (d,
3H), 3.76 (s, 3H), 3.85 (dd, 1H), 4.00 (dd, 1H), 4.30 (br. s, 1H),
4.52 (quin, 1H), 4.96-5.11 (m, 2H), 5.37 (d, 1H), 6.90 (d, 1H),
7.09 (td, 1H), 7.24 (d, 1H), 7.35 (dd, 1H), 7.52 (td, 1H),
7.60-7.65 (m, 2H), 7.73-7.79 (m, 2H).
Example 12
5-(4-Chlorophenyl)-2-{[5-(1-hydroxyethyl)-1-(2-methoxyphenyl)-1H-1,2,4-tri-
azol-3-yl]methyl}-4-[(2S)-3,3,3-trifluoro-2-hydroxypropyl]-2,4-dihydro-3H--
1,2,4-triazol-3-one (diastereomer 2)
[0277] Analytical chiral HPLC (SFC): R.sub.t=3.38 min, d.e.=91.1%
[column: Daicel Chiralcel.RTM. OX-3 250.times.4 mm; eluent: carbon
dioxide/methanol (5%.fwdarw.60%); flow rate: 3 ml/min; UV
detection: 220 nm].
[0278] .sup.1H NMR (400 MHz, DMSO-d.sub.6): .delta. [ppm] 1.36 (d,
3H), 3.76 (s, 3H), 3.84 (dd, 1H), 4.00 (dd, 1H), 4.31 (br. s, 1H),
4.52 (quin, 1H), 4.99-5.09 (m, 2H), 5.37 (d, 1H), 6.91 (d, 1H),
7.09 (td, 1H), 7.24 (d, 1H), 7.35 (dd, 1H), 7.52 (td, 1H),
7.60-7.65 (m, 2H), 7.74-7.79 (m, 2H).
Example 13
2-({1-(3-Chloro-5-fluorophenyl)-5-[(1RS)-1-hydroxyethyl]-1H-1,2,4-triazol--
3-yl}methyl)-5-(4-chlorophenyl)-4-[(2S)-3,3,3-trifluoro-2-hydroxypropyl]-2-
,4-dihydro-3H-1,2,4-triazol-3-one (diastereomeric mixture)
##STR00029##
[0280] To a solution of
5-(4-chlorophenyl)-2-({5-[(1RS)-1-hydroxyethyl]-1H-1,2,4-triazol-3-yl}met-
hyl)-4-[(2S)-3,3,3-trifluoro-2-hydroxypropyl]-2,4-dihydro-3H-1,2,4-triazol-
-3-one (430 mg, 0.99 mmol) in pyridine (12.5 ml) were added
(3-chloro-5-fluorophenyl)boronic acid (346.49 mg, 1.99 mmol) and
copper(II) acetate (360.9 mg, 1.99 mmol). The reaction mixture was
stirred at room temperature for 5 days, after which extra boronic
acid (86.7 mg, 0.497 mmol, 0.5 eq.) was added due to incomplete
conversion. After stirring for two additional days, the reaction
mixture was concentrated in vacuo, then diluted with MTBE and
quenched with aqueous hydrochloric acid (0.5 M). After phase
separation, the aqueous phase was extracted twice with MTBE. The
combined organic phases were dried over sodium sulfate, filtered,
and concentrated in vacuo. The crude product was purified by
preparative HPLC [method 4], and the desired compound (148 mg, 0.26
mmol) was obtained as a mixture of diastereomers (yield 26.5%).
[0281] LC/MS [method 2]: R.sub.t=3.28 min; MS [ESIpos]: m/z=561
(M+H).sup.+
[0282] .sup.1H NMR (400 MHz, DMSO-d.sub.6): .delta. [ppm] 1.48 (d,
3H), 3.85 (dd, 1H), 4.01 (dd, 1H), 4.29 (br. s, 1H), 4.83-4.91 (m,
1H), 5.01-5.13 (m, 2H), 6.89 (br. s, 1H), 7.55-7.70 (m, 5H),
7.72-7.78 (m, 2H).
[0283] The two diastereomers were separated by preparative chiral
HPLC (SFC) [sample preparation: 143 mg dissolved in 15 ml methanol;
injection volume: 0.5 ml; column: Daicel Chiralcel.RTM. OX-H 5
.mu.m, 250.times.20 mm; eluent: carbon dioxide/methanol 80:20; flow
rate: 80 ml/min; temperature: 40.degree. C.; UV detection: 210 nm].
After separation, 70 mg of diastereomer 1 (Example 14), which
eluted first, and 60 mg of diastereomer 2 (Example 15), which
eluted later, were isolated.
Example 14
2-{[1-(3-Chloro-5-fluorophenyl)-5-(1-hydroxyethyl)-1H-H-1,2,4-triazol-3-yl-
]methyl}-5-(4-chlorophenyl)-4-[(2S)-3,3,3-trifluoro-2-hydroxypropyl]-2,4-d-
ihydro-3H-1,2,4-triazol-3-one (diastereomer 1)
[0284] Analytical chiral HPLC (SFC): R.sub.t=4.45 min, d.e.=100%
[column: Daicel Chiralcel.RTM. OX-3 250.times.4 mm; eluent: carbon
dioxide/methanol (5%.fwdarw.60%); flow rate: 3 ml/min; UV
detection: 220 nm].
[0285] .sup.1H NMR (400 MHz, DMSO-d.sub.6): .delta. [ppm] 1.48 (d,
3H), 3.85 (dd, 1H), 4.01 (dd, 1H), 4.24-4.36 (m, 1H), 4.87 (quin,
1H), 5.07 (s, 2H), 5.83 (d, 1H), 6.89 (d, 1H), 7.58-7.69 (m, 5H),
7.71-7.79 (m, 2H).
Example 15
2-{[1-(3-Chloro-5-fluorophenyl)-5-(1-hydroxyethyl)-1H-H-1,2,4-triazol-3-yl-
]methyl}-5-(4-chlorophenyl)-4-[(2S)-3,3,3-trifluoro-2-hydroxypropyl]-2,4-d-
ihydro-3H-1,2,4-triazol-3-one (diastereomer 2)
[0286] Analytical chiral HPLC (SFC): R.sub.t=4.80 min, d.e.=100%
[column: Daicel Chiralcel.RTM. OX-3 250.times.4 mm; eluent: carbon
dioxide/methanol (5%.fwdarw.60%); flow rate: 3 ml/min; UV
detection: 220 nm].
[0287] .sup.1H NMR (400 MHz, DMSO-d.sub.6): .delta. [ppm] 1.48 (d,
3H), 3.85 (dd, 1H), 4.01 (dd, 1H), 4.23-4.36 (m, 1H), 4.87 (quin,
1H), 5.02-5.12 (m, 2H), 5.83 (d, 1H), 6.89 (d, 1H), 7.57-7.70 (m,
5H), 7.71-7.79 (m, 2H).
Example 16
5-(4-Chlorophenyl)-2-({5-[(1RS)-1-hydroxyethyl]-1-phenyl-1H-1,2,4-triazol--
3-yl}methyl)-4-[(2S)-3,3,3-trifluoro-2-hydroxypropyl]-2,4-dihydro-3H-1,2,4-
-triazol-3-one (diastereomeric mixture)
##STR00030##
[0289] To a solution of
5-(4-chlorophenyl)-2-({5-[(1RS)-1-hydroxyethyl]-1H-1,2,4-triazol-3-yl}met-
hyl)-4-[(2S)-3,3,3-trifluoro-2-hydroxypropyl]-2,4-dihydro-3H-1,2,4-triazol-
-3-one (250 mg, 0.462 mmol, 80% purity) in pyridine (6 ml) were
added phenylboronic acid (112.69 mg, 0.92 mmol) and copper(II)
acetate (167.9 mg, 0.92 mmol). The reaction mixture was heated to
60.degree. C. for 2 h and then stirred at room temperature for 4
days, after which extra boronic acid (28.2 mg, 0.23 mmol, 0.5 eq.)
was added due to incomplete conversion. After further 4 h at
60.degree. C. followed by stirring at room temperature for two
additional days, the reaction mixture was concentrated in vacuo,
then diluted with ethyl acetate and quenched with aqueous
hydrochloric acid (0.5 M). After phase separation, the aqueous
phase was extracted twice with ethyl acetate. The combined organic
phases were dried over sodium sulfate, filtered, and concentrated
in vacuo. The crude product was purified by preparative HPLC
[method 4], and the desired compound (42.5 mg, 0.08 mmol) was
obtained as a mixture of diastereomers (yield 18.1%).
[0290] LC/MS [method 3]: R.sub.t=1.21 min; MS [ESIpos]: m/z=509
(M+H).sup.+
[0291] .sup.1H NMR (400 MHz, DMSO-d.sub.6): .delta. [ppm] 1.45 (d,
3H), 3.85 (dd, 1H), 4.00 (dd, 1H), 4.30 (br. s, 1H), 4.77 (q, 1H),
4.99-5.13 (m, 2H), 6.90 (br. s, 1H), 7.47-7.66 (m, 7H), 7.72-7.79
(m, 2H).
[0292] The two diastereomers were separated by preparative chiral
HPLC [sample preparation: 38.9 mg dissolved in 1 ml
ethanol/isohexane (1:1); injection volume: 1 ml; column: Daicel
Chiralcel.RTM. OX-H 5 .mu.m, 250.times.20 mm; eluent:
isohexane/ethanol 75:25; flow rate: 15 ml/min; temperature:
30.degree. C.; UV detection: 220 nm]. After separation, 13 mg of
diastereomer 1 (Example 17), which eluted first, and 14 mg of
diastereomer 2 (Example 18), which eluted later, were isolated.
Example 17
5-(4-Chlorophenyl)-2-{[5-(1-hydroxyethyl)-1-phenyl-1H-1,2,4-triazol-3-yl]m-
ethyl}-4-[(2S)-3,3,3-trifluoro-2-hydroxypropyl]-2,4-dihydro-3H-1,2,4-triaz-
ol-3-one (diastereomer 1)
[0293] Analytical chiral HPLC: R.sub.t=8.18 min, d.e.=100% [column:
LUX Cellulose-4, 5 .mu.m, 250.times.4.6 mm; eluent:
isohexane/ethanol 70:30; flow rate: 1 ml/min; temperature:
40.degree. C.; UV detection: 220 nm].
[0294] .sup.1H NMR (400 MHz, DMSO-d.sub.6): .delta. [ppm] 1.45 (d,
3H), 3.85 (dd, 1H), 4.00 (dd, 1H), 4.30 (br. s, 1H), 4.77 (quin,
1H), 4.98-5.14 (m, 2H), 5.68 (d, 1H), 6.89 (d, 1H), 7.48-7.65 (m,
7H), 7.72-7.80 (m, 2H).
Example 18
5-(4-Chlorophenyl)-2-{[5-(1-hydroxyethyl)-1-phenyl-1H-1,2,4-triazol-3-yl]m-
ethyl}-4-[(2S)-3,3,3-trifluoro-2-hydroxypropyl]-2,4-dihydro-3H-1,2,4-triaz-
ol-3-one (diastereomer 2)
[0295] Analytical chiral HPLC: R.sub.t=11.40 min, d.e.=100%
[column: LUX Cellulose-4, 5 .mu.m, 250.times.4.6 mm; eluent:
isohexane/ethanol 70:30; flow rate: 1 ml/min; temperature:
40.degree. C.; UV detection: 220 nm].
[0296] .sup.1H NMR (400 MHz, DMSO-d.sub.6): .delta. [ppm] 1.45 (d,
3H), 3.85 (dd, 1H), 4.00 (dd, 1H), 4.30 (br. s, 1H), 4.77 (quin,
1H), 5.07 (s, 2H), 5.68 (d, 1H), 6.90 (d, 1H), 7.47-7.65 (m, 7H),
7.72-7.79 (m, 2H).
Example 19
5-(4-Chlorophenyl)-2-({1-[3-(difluoromethyl)phenyl]-5-[(1RS)-1-hydroxyethy-
l]-1H-1,2,4-triazol-3-yl}methyl)-4-[(2S)-3,3,3-trifluoro-2-hydroxypropyl]--
2,4-dihydro-3H-1,2,4-triazol-3-one (diastereomeric mixture)
##STR00031##
[0298] To a solution of
5-(4-chlorophenyl)-2-({5-[(1RS)-1-hydroxyethyl]-1H-1,2,4-triazol-3-yl}met-
hyl)-4-[(2S)-3,3,3-trifluoro-2-hydroxypropyl]-2,4-dihydro-3H-1,2,4-triazol-
-3-one (400 mg, 0.74 mmol, 80% purity) in pyridine (9.6 ml) were
added [3-(difluoromethyl)phenyl]boronic acid (254.26 mg, 1.48 mmol)
and copper(II) acetate (268.6 mg, 1.48 mmol). The reaction mixture
was heated to 60.degree. C. for 2 h and then stirred at room
temperature for 5 days. The resulting reaction mixture was
concentrated in vacuo, then diluted with ethyl acetate and quenched
with aqueous hydrochloric acid (0.5 M). After phase separation, the
aqueous phase was extracted twice with ethyl acetate. The combined
organic phases were dried over sodium sulfate, filtered, and
concentrated in vacuo. The crude product was purified by
preparative HPLC [method 4], and the desired compound (47 mg, 0.08
mmol) was obtained as a mixture of diastereomers (yield 11.3%).
[0299] LC/MS [method 1]: R.sub.t=1.04 min; MS [ESIpos]: m/z=559
(M+H).sup.+
[0300] .sup.1H NMR (400 MHz, DMSO-d.sub.6): .delta. [ppm] 1.47 (d,
3H), 3.85 (dd, 1H), 4.01 (dd, 1H), 4.24-4.36 (m, 1H), 4.81 (q, 1H),
5.02-5.13 (m, 2H), 5.74 (br. s, 1H), 6.89 (br. s, 1H), 7.14 (t,
1H), 7.59-7.65 (m, 2H), 7.69-7.78 (m, 4H), 7.81-7.87 (m, 2H).
[0301] The two diastereomers were separated by preparative chiral
HPLC [sample preparation: 45 mg dissolved in 1 ml ethanol/isohexane
(1:1); injection volume: 1 ml; column: Daicel Chiralcel.RTM. OX-H 5
.mu.m, 250.times.20 mm; eluent: isohexane/ethanol 75:25; flow rate:
15 ml/min; temperature: 30.degree. C.; UV detection: 220 nm]. After
separation, 20 mg of diastereomer 1 (Example 20), which eluted
first, and 20 mg of diastereomer 2 (Example 21), which eluted
later, were isolated.
Example 20
5-(4-Chlorophenyl)-2-({1-[3-(difluoromethyl)phenyl]-5-(1-hydroxyethyl)-1H--
1,2,4-triazol-3-yl}-methyl)-4-[(2S)-3,3,3-trifluoro-2-hydroxypropyl]-2,4-d-
ihydro-3H-1,2,4-triazol-3-one (diastereomer 1)
[0302] Analytical chiral HPLC: R.sub.t=6.72 min, d.e.=99% [column:
LUX Cellulose-4, 5 .mu.m, 250.times.4.6 mm; eluent:
isohexane/ethanol 70:30; flow rate: 1 ml/min; temperature:
40.degree. C.; UV detection: 220 nm].
[0303] .sup.1H NMR (400 MHz, DMSO-d.sub.6): .delta. [ppm] 1.47 (d,
3H), 3.85 (dd, 1H), 4.01 (dd, 1H), 4.23-4.36 (m, 1H), 4.81 (quin,
1H), 5.02-5.14 (m, 2H), 5.74 (d, 1H), 6.88 (d, 1H), 7.14 (t, 1H),
7.59-7.64 (m, 2H), 7.69-7.79 (m, 4H), 7.80-7.87 (m, 2H).
Example 21
5-(4-Chlorophenyl)-2-({1-[3-(difluoromethyl)phenyl]-5-(1-hydroxyethyl)-1H--
1,2,4-triazol-3-yl}-methyl)-4-[(2S)-3,3,3-trifluoro-2-hydroxypropyl]-2,4-d-
ihydro-3H-1,2,4-triazol-3-one (diastereomer 2)
[0304] Analytical chiral HPLC: R.sub.t=9.36 min, d.e.=100% [column:
LUX Cellulose-4, 5 .mu.m, 250.times.4.6 mm; eluent:
isohexane/ethanol 70:30; flow rate: 1 ml/min; temperature:
40.degree. C.; UV detection: 220 nm].
[0305] .sup.1H NMR (400 MHz, DMSO-d.sub.6): .delta. [ppm] 1.47 (d,
3H), 3.85 (dd, 1H), 4.01 (dd, 1H), 4.24-4.37 (m, 1H), 4.81 (quin,
1H), 5.08 (s, 2H), 5.74 (d, 1H), 6.89 (d, 1H), 7.13 (t, 1H),
7.58-7.66 (m, 2H), 7.69-7.79 (m, 4H), 7.81-7.87 (m, 2H).
Example 22
5-(4-Chlorophenyl)-2-({5-[(RS)-1-hydroxyethyl]-1-[3-(trifluoromethyl)pheny-
l]-H-1,2,4-triazol-3-yl}methyl)-4-[(2S)-3,3,3-trifluoro-2-hydroxypropyl]-2-
,4-dihydro-3H-1,2,4-triazol-3-one (diastereomeric mixture)
##STR00032##
[0307] To a solution of
5-(4-chlorophenyl)-2-({5-[(1RS)-1-hydroxyethyl]-1H-1,2,4-triazol-3-yl}met-
hyl)-4-[(2S)-3,3,3-trifluoro-2-hydroxypropyl]-2,4-dihydro-3H-1,2,4-triazol-
-3-one (400 mg, 0.74 mmol, 80% purity) in pyridine (9.6 ml) were
added [3-(trifluoromethyl)phenyl]boronic acid (280.86 mg, 1.48
mmol) and copper(II) acetate (268.6 mg, 1.48 mmol). The reaction
mixture was heated to 60.degree. C. for 2 h and then stirred at
room temperature for 5 days, after which extra boronic acid (70.2
mg, 0.37 mmol, 0.5 eq.) was added due to incomplete conversion.
After further 4 h at 60.degree. C. followed by stirring at room
temperature overnight, the reaction mixture was concentrated in
vacuo, then diluted with MTBE and quenched with aqueous
hydrochloric acid (0.5 M). After phase separation, the aqueous
phase was extracted twice with MTBE. The combined organic phases
were dried over sodium sulfate, filtered, and concentrated in
vacuo. The crude product was purified by preparative HPLC [method
4], and the desired compound (58.2 mg, 0.10 mmol) was obtained as a
mixture of diastereomers (yield 13.6%).
[0308] LC/MS [method 1]: R.sub.t=1.06 min; MS [ESIpos]: m/z=577
(M+H).sup.+
[0309] .sup.1H NMR (400 MHz, DMSO-d.sub.6): .delta. [ppm] 1.48 (d,
3H), 3.85 (dd, 1H), 4.01 (dd, 1H), 4.30 (br. s, 1H), 4.83 (q, 1H),
5.02-5.15 (m, 2H), 5.79 (br. s, 1H), 6.85-6.94 (m, 1H), 7.58-7.66
(m, 2H), 7.71-7.85 (m, 3H), 7.86-7.92 (m, 1H), 7.94-8.01 (m, 1H),
8.04 (s, 1H).
[0310] The two diastereomers were separated by preparative chiral
HPLC [sample preparation: 58 mg dissolved in 2 ml ethanol/isohexane
(1:1); injection volume: 1 ml; column: Daicel Chiralcel.RTM. OX-H
m, 250.times.20 mm; eluent: isohexane/ethanol 75:25; flow rate: 15
ml/min; temperature: 30.degree. C.; UV detection: 220 nm]. After
separation, 19.5 mg of diastereomer 1 (Example 23), which eluted
first, and 19.2 mg of diastereomer 2 (Example 24), which eluted
later, were isolated.
Example 23
5-(4-Chlorophenyl)-2-({5-(1-hydroxyethyl)-1-[3-(trifluoromethyl)phenyl]-H--
1,2,4-triazol-3-yl}-methyl)-4-[(2S)-3,3,3-trifluoro-2-hydroxypropyl]-2,4-d-
ihydro-3H-1,2,4-triazol-3-one (diastereomer 1)
[0311] Analytical chiral HPLC: R.sub.t=4.94 min, d.e.=100% [column:
LUX Cellulose-4, 5 .mu.m, 250.times.4.6 mm; eluent:
isohexane/ethanol 70:30; flow rate: 1 ml/min; temperature:
40.degree. C.; UV detection: 220 nm].
[0312] .sup.1H NMR (400 MHz, DMSO-d.sub.6): .delta. [ppm] 1.48 (d,
3H), 3.85 (dd, 1H), 4.01 (dd, 1H), 4.30 (br. s, 1H), 4.83 (q, 1H),
5.03-5.13 (m, 2H), 5.79 (br. s, 1H), 6.88 (d, 1H), 7.59-7.65 (m,
2H), 7.72-7.85 (m, 3H), 7.86-7.92 (m, 1H), 7.95-8.01 (m, 1H), 8.04
(br. s, 1H).
Example 24
5-(4-Chlorophenyl)-2-({5-(1-hydroxyethyl)-1-[3-(trifluoromethyl)phenyl]-H--
1,2,4-triazol-3-yl}-methyl)-4-[(2S)-3,3,3-trifluoro-2-hydroxypropyl]-2,4-d-
ihydro-3H-1,2,4-triazol-3-one (diastereomer 2)
[0313] Analytical chiral HPLC: R.sub.t=6.13 min, d.e.=98.6%
[column: LUX Cellulose-4, 5 .mu.m, 250.times.4.6 mm; eluent:
isohexane/ethanol 70:30; flow rate: 1 ml/min; temperature:
40.degree. C.; UV detection: 220 nm].
[0314] .sup.1H NMR (400 MHz, DMSO-d.sub.6): .delta. [ppm] 1.48 (d,
3H), 3.85 (dd, 1H), 4.01 (dd, 1H), 4.24-4.36 (m, 1H), 4.83 (quin,
1H), 5.09 (s, 2H), 5.78 (d, 1H), 6.89 (d, 1H), 7.62 (d, 2H),
7.71-7.77 (m, 2H), 7.78-7.85 (m, 1H), 7.86-7.92 (m, 1H), 7.94-8.01
(m, 1H), 8.04 (br. s, 1H).
Example 25
5-(4-Chlorophenyl)-2-({1-(3,5-difluorophenyl)-5-[(1RS)-1-hydroxyethyl]-1H--
1,2,4-triazol-3-yl}-methyl)-4-[(2S)-3,3,3-trifluoro-2-hydroxypropyl]-2,4-d-
ihydro-3H-1,2,4-triazol-3-one (diastereomeric mixture)
##STR00033##
[0316] To a solution of
5-(4-chlorophenyl)-2-({5-[(1RS)-1-hydroxyethyl]-1H-1,2,4-triazol-3-yl}met-
hyl)-4-[(2S)-3,3,3-trifluoro-2-hydroxypropyl]-2,4-dihydro-3H-1,2,4-triazol-
-3-one (400 mg, 0.92 mmol) in pyridine (12 ml) were added
(3,5-difluorophenyl)boronic acid (291.9 mg, 1.85 mmol) and
copper(II) acetate (335.7 mg, 1.85 mmol). The reaction mixture was
heated to 60.degree. C. for 2 h and then stirred at room
temperature for 4 days, after which extra boronic acid (72.98 mg,
0.46 mmol, 0.5 eq.) was added due to incomplete conversion. After
further 2 h at 60.degree. C. followed by stirring at room
temperature for three additional days, the reaction mixture was
concentrated in vacuo, then diluted with ethyl acetate and quenched
with aqueous hydrochloric acid (0.5 M). After phase separation, the
aqueous phase was extracted twice with ethyl acetate. The combined
organic phases were dried over sodium sulfate, filtered, and
concentrated in vacuo. The crude product was purified by
preparative HPLC [method 4], and the desired compound (114.3 mg,
0.21 mmol) was obtained as a mixture of diastereomers (yield
22.7%).
[0317] LC/MS [method 3]: R.sub.t=1.28 min; MS [ESIpos]: m/z=545
(M+H).sup.+
[0318] .sup.1H NMR (400 MHz, DMSO-d.sub.6): .delta. [ppm] 1.49 (d,
3H), 3.85 (dd, 1H), 4.01 (dd, 1H), 4.30 (br. s, 1H), 4.88 (q, 1H),
5.02-5.13 (m, 2H), 6.89 (br. s, 1H), 7.41-7.54 (m, 3H), 7.62 (d,
2H), 7.75 (d, 2H).
[0319] The two diastereomers were separated by preparative chiral
HPLC [sample preparation: 110 mg dissolved in 7 ml
ethanol/isohexane (1:1); injection volume: 0.6 ml; column: Daicel
Chiralcel.RTM. OX-H 5 .mu.m, 250.times.20 mm; eluent:
isohexane/ethanol 70:30; flow rate: 20 ml/min; temperature:
40.degree. C.; UV detection: 220 nm]. After separation, 42 mg of
diastereomer 1 (Example 26), which eluted first, and 44 mg of
diastereomer 2 (Example 27), which eluted later, were isolated.
Example 26
5-(4-Chlorophenyl)-2-{[1-(3,5-difluorophenyl)-5-(1-hydroxyethyl)-1H-1,2,4--
triazol-3-yl]methyl}-4-[(2S)-3,3,3-trifluoro-2-hydroxypropyl]-2,4-dihydro--
3H-1,2,4-triazol-3-one (diastereomer 1)
[0320] LC/MS [method 2]: R.sub.t=3.10 min; MS [ESIpos]: m/z=545
(M+H).sup.+
[0321] Analytical chiral HPLC: R.sub.t=1.09 min, d.e.=100% [column:
Daicel Chiralpack OX-3 3 .mu.m, 50.times.4.6 mm; eluent:
isohexane/ethanol 70:30; flow rate: 1 ml/min; UV detection: 220
nm].
[0322] .sup.1H NMR (400 MHz, DMSO-d.sub.6): .delta. [ppm] 1.49 (d,
3H), 3.85 (dd, 1H), 4.01 (dd, 1H), 4.23-4.36 (m, 1H), 4.88 (quin,
1H), 5.02-5.13 (m, 2H), 5.84 (d, 1H), 6.89 (d, 1H), 7.41-7.54 (m,
3H), 7.59-7.65 (m, 2H), 7.73-7.76 (m, 2H).
Example 27
5-(4-Chlorophenyl)-2-{[1-(3,5-difluorophenyl)-5-(1-hydroxyethyl)-1H-1,2,4--
triazol-3-yl]methyl}-4-[(2S)-3,3,3-trifluoro-2-hydroxypropyl]-2,4-dihydro--
3H-1,2,4-triazol-3-one (diastereomer 2)
[0323] LC/MS [method 2]: R.sub.t=3.09 min; MS [ESIpos]: m/z=545
(M+H).sup.+
[0324] Analytical chiral HPLC: R.sub.t=1.28 min, d.e.=99% [column:
Daicel Chiralpack OX-3 3 .mu.m, 50.times.4.6 mm; eluent:
isohexane/ethanol 70:30; flow rate: 1 ml/min; UV detection: 220
nm].
[0325] .sup.1H NMR (400 MHz, DMSO-d.sub.6): .delta. [ppm] 1.48 (d,
3H), 3.85 (dd, 1H), 4.01 (dd, 1H), 4.24-4.36 (m, 1H), 4.88 (quin,
1H), 5.07 (s, 2H), 5.83 (d, 1H), 6.90 (d, 1H), 7.42-7.54 (m, 3H),
7.59-7.65 (m, 2H), 7.72-7.79 (m, 2H).
Example 28
5-(4-Chlorophenyl)-2-({5-[(1RS)-1-hydroxyethyl]-1-(3-methylphenyl)-1H-1,2,-
4-triazol-3-yl}-methyl)-4-[(2S)-3,3,3-trifluoro-2-hydroxypropyl]-2,4-dihyd-
ro-3H-1,2,4-triazol-3-one (diastereomeric mixture)
##STR00034##
[0327] To a solution of
5-(4-chlorophenyl)-2-({5-[(1RS)-1-hydroxyethyl]-1H-1,2,4-triazol-3-yl}met-
hyl)-4-[(2S)-3,3,3-trifluoro-2-hydroxypropyl]-2,4-dihydro-3H-1,2,4-triazol-
-3-one (400 mg, 0.92 mmol) in pyridine (12 ml) were added
(3-methylphenyl)boronic acid (251.32 mg, 1.85 mmol) and copper(II)
acetate (335.7 mg, 1.85 mmol). The reaction mixture was heated to
60.degree. C. for 2 h and then stirred at room temperature for
three days. The resulting reaction mixture was concentrated in
vacuo, then diluted with ethyl acetate and quenched with aqueous
hydrochloric acid (0.5 M). After phase separation, the aqueous
phase was extracted twice with ethyl acetate. The combined organic
phases were dried over sodium sulfate, filtered, and concentrated
in vacuo. The crude product was purified by preparative HPLC
[method 4], and the desired compound (59.6 mg, 0.11 mmol) was
obtained as a mixture of diastereomers (yield 12.3%).
[0328] .sup.1H NMR (400 MHz, DMSO-d.sub.6): .delta. [ppm] 1.44 (d,
3H), 2.38 (s, 3H), 3.85 (dd, 1H), 4.00 (dd, 1H), 4.29 (br. s, 1H),
4.76 (q, 1H), 5.01-5.11 (m, 2H), 5.66 (br. s, 1H), 6.90 (t, 1H),
7.29-7.35 (m, 1H), 7.38-7.47 (m, 3H), 7.59-7.65 (m, 2H), 7.72-7.78
(m, 2H).
[0329] The two diastereomers were separated by preparative chiral
HPLC [sample preparation: 56 mg dissolved in 2 ml ethanol/isohexane
(1:1); injection volume: 0.5 ml; column: Daicel Chiralcel.RTM. OX-H
5 .mu.m, 250.times.20 mm; eluent: isohexane/ethanol 70:30; flow
rate: 15 ml/min; temperature: 25.degree. C.; UV detection: 220 nm].
After separation, 22 mg of diastereomer 1 (Example 29), which
eluted first, and 24 mg of diastereomer 2 (Example 30), which
eluted later, were isolated.
Example 29
5-(4-Chlorophenyl)-2-{[5-(1-hydroxyethyl)-1-(3-methylphenyl)-1H-1,2,4-tria-
zol-3-yl]methyl}-4-[(2S)-3,3,3-trifluoro-2-hydroxypropyl]-2,4-dihydro-3H-1-
,2,4-triazol-3-one (diastereomer 1)
[0330] Analytical chiral HPLC: R.sub.t=7.97 min, d.e.=100% [column:
LUX Cellulose-4, 5 .mu.m, 250.times.4.6 mm; eluent:
isohexane/ethanol 70:30; flow rate: 1 ml/min; temperature:
40.degree. C.; UV detection: 220 nm].
[0331] .sup.1H NMR (400 MHz, DMSO-d.sub.6): .delta. [ppm] 1.44 (d,
3H), 2.38 (s, 3H), 3.85 (dd, 1H), 4.01 (dd, 1H), 4.24-4.36 (m, 1H),
4.76 (quin, 1H), 5.00-5.11 (m, 2H), 5.67 (d, 1H), 6.89 (d, 1H),
7.30-7.35 (m, 1H), 7.38-7.47 (m, 3H), 7.59-7.65 (m, 2H), 7.72-7.78
(m, 2H).
Example 30
5-(4-Chlorophenyl)-2-{[5-(1-hydroxyethyl)-1-(3-methylphenyl)-1H-1,2,4-tria-
zol-3-yl]methyl}-4-[(2S)-3,3,3-trifluoro-2-hydroxypropyl]-2,4-dihydro-3H-1-
,2,4-triazol-3-one (diastereomer 2)
[0332] Analytical chiral HPLC: R.sub.t=11.44 min, d.e.=99.1%
[column: LUX Cellulose-4, 5 .mu.m, 250.times.4.6 mm; eluent:
isohexane/ethanol 70:30; flow rate: 1 ml/min; temperature:
40.degree. C.; UV detection: 220 nm].
[0333] .sup.1H NMR (400 MHz, DMSO-d.sub.6): .delta. [ppm] 1.44 (d,
3H), 2.38 (s, 3H), 3.85 (dd, 1H), 4.00 (dd, 1H), 4.24-4.36 (m, 1H),
4.77 (quin, 1H), 5.01-5.10 (m, 2H), 5.67 (d, 1H), 6.90 (d, 1H),
7.29-7.35 (m, 1H), 7.38-7.47 (m, 3H), 7.59-7.65 (m, 2H), 7.73-7.78
(m, 2H).
Example 31
5-(4-Chlorophenyl)-2-({1-(2-ethylphenyl)-5-[(1RS)-1-hydroxyethyl]-1H-1,2,4-
-triazol-3-yl}-methyl)-4-[(2S)-3,3,3-trifluoro-2-hydroxypropyl]-2,4-dihydr-
o-3H-1,2,4-triazol-3-one (diastereomeric mixture)
##STR00035##
[0335] To a solution of
5-(4-chlorophenyl)-2-({5-[(1RS)-1-hydroxyethyl]-1H-1,2,4-triazol-3-yl}met-
hyl)-4-[(2S)-3,3,3-trifluoro-2-hydroxypropyl]-2,4-dihydro-3H-1,2,4-triazol-
-3-one (600 mg, 1.39 mmol) in pyridine (18 ml) were added
(2-ethylphenyl)boronic acid (415.87 mg, 2.77 mmol) and copper(II)
acetate (503.6 mg, 2.77 mmol). The reaction mixture was heated to
60.degree. C. for 2 h and then stirred at room temperature for
three days. The resulting reaction mixture was concentrated in
vacuo, then diluted with ethyl acetate and quenched with aqueous
hydrochloric acid (0.5 M). After phase separation, the aqueous
phase was extracted twice with ethyl acetate. The combined organic
phases were dried over sodium sulfate, filtered, and concentrated
in vacuo. The crude product was purified by preparative HPLC
[method 4], and the desired compound (69.4 mg, 0.13 mmol) was
obtained as a mixture of diastereomers (yield 9.1%).
[0336] LC/MS [method 2]: R.sub.t=3.16 min; MS [ESIpos]: m/z=537
(M+H).sup.+
[0337] .sup.1H NMR (400 MHz, DMSO-d.sub.6): .delta. [ppm] 0.98 (t,
3H), 1.37 (d, 3H), 2.27 (qd, 2H), 3.84 (dd, 1H), 4.00 (dd, 1H),
4.21-4.37 (m, 1H), 4.52 (q, 1H), 5.00-5.13 (m, 2H), 5.48 (br. s,
1H), 6.90 (dd, 1H), 7.32-7.40 (m, 2H), 7.41-7.54 (m, 2H), 7.58-7.65
(m, 2H), 7.70-7.77 (m, 2H).
[0338] The two diastereomers were separated by preparative chiral
HPLC [sample preparation: 65 mg dissolved in 4 ml ethanol/isohexane
(1:1); injection volume: 0.5 ml; column: Daicel Chiralcel.RTM. OX-H
5 .mu.m, 250.times.20 mm; eluent: isohexane/ethanol 50:50; flow
rate: 20 ml/min; temperature: 40.degree. C.; UV detection: 220 nm].
After separation, 25 mg of diastereomer 1 (Example 32), which
eluted first, and 25 mg of diastereomer 2 (Example 33), which
eluted later, were isolated.
Example 32
5-(4-Chlorophenyl)-2-{[1-(2-ethylphenyl)-5-(1-hydroxyethyl)-1H-1,2,4-triaz-
ol-3-yl]methyl}-4-[(2S)-3,3,3-trifluoro-2-hydroxypropyl]-2,4-dihydro-3H-1,-
2,4-triazol-3-one (diastereomer 1)
[0339] LC/MS [method 2]: R.sub.t=3.15 min; MS [ESIpos]: m/z=537
(M+H).sup.+
[0340] Analytical chiral HPLC: R.sub.t=0.96 min, d.e.=100% [column:
Daicel Chiralpack OX-3 3 .mu.m, 50.times.4.6 mm; eluent:
isohexane/ethanol 50:50; flow rate: 1 ml/min; UV detection: 220
nm].
[0341] .sup.1H NMR (400 MHz, DMSO-d.sub.6): .delta. [ppm] 0.98 (t,
3H), 1.37 (d, 3H), 2.27 (qd, 2H), 3.84 (dd, 1H), 4.00 (dd, 1H),
4.23-4.35 (m, 1H), 4.52 (quin, 1H), 5.00-5.12 (m, 2H), 5.48 (d,
1H), 6.89 (d, 1H), 7.32-7.40 (m, 2H), 7.42-7.53 (m, 2H), 7.59-7.66
(m, 2H), 7.71-7.78 (m, 2H).
Example 33
5-(4-Chlorophenyl)-2-{[1-(2-ethylphenyl)-5-(1-hydroxyethyl)-1H-1,2,4-triaz-
ol-3-yl]methyl}-4-[(2S)-3,3,3-trifluoro-2-hydroxypropyl]-2,4-dihydro-3H-1,-
2,4-triazol-3-one (diastereomer 2)
[0342] LC/MS [method 2]: R.sub.t=3.15 min; MS [ESIpos]: m/z=537
(M+H).sup.+
[0343] Analytical chiral HPLC: R.sub.t=1.09 min, d.e.=100% [column:
Daicel Chiralpack OX-3 3 .mu.m, 50.times.4.6 mm; eluent:
isohexane/ethanol 50:50; flow rate: 1 ml/min; UV detection: 220
nm].
[0344] .sup.1H NMR (400 MHz, DMSO-d.sub.6): .delta. [ppm] 0.98 (t,
3H), 1.37 (d, 3H), 2.27 (qd, 2H), 3.84 (dd, 1H), 4.00 (dd, 1H),
4.23-4.36 (m, 1H), 4.52 (quin, 1H), 5.07 (s, 2H), 5.48 (d, 1H),
6.90 (d, 1H), 7.32-7.40 (m, 2H), 7.42-7.54 (m, 2H), 7.59-7.66 (m,
2H), 7.71-7.78 (m, 2H).
Example 34
2-({1-(2-Chloro-4-fluorophenyl)-5-[(1RS)-1-hydroxyethyl]-1H-1,2,4-triazol--
3-yl}methyl)-5-(4-chlorophenyl)-4-[(2S)-3,3,3-trifluoro-2-hydroxypropyl]-2-
,4-dihydro-3H-1,2,4-triazol-3-one (diastereomeric mixture)
##STR00036##
[0346] To a solution of
5-(4-chlorophenyl)-2-({5-[(1RS)-1-hydroxyethyl]-1H-1,2,4-triazol-3-yl}met-
hyl)-4-[(2S)-3,3,3-trifluoro-2-hydroxypropyl]-2,4-dihydro-3H-1,2,4-triazol-
-3-one (600 mg, 1.39 mmol) in pyridine (18 ml) were added
(2-chloro-4-fluorophenyl)boronic acid (483 mg, 2.77 mmol) and
copper(II) acetate (503.6 mg, 2.77 mmol). The reaction mixture was
heated to 60.degree. C. for 2 h and then stirred at room
temperature for 5 days, after which extra boronic acid (242 mg,
1.39 mmol) was added due to incomplete conversion. The reaction
mixture was further stirred at room temperature for 4 days. Over
this time, two additional portions of boronic acid (483 mg in
total, 2.77 mmol) were added. After this, the reaction mixture was
concentrated in vacuo, then diluted with MTBE and quenched with
aqueous hydrochloric acid (0.5 M). After phase separation, the
aqueous phase was extracted twice with MTBE. The combined organic
phases were dried over sodium sulfate, filtered, and concentrated
in vacuo. The crude product was purified by preparative HPLC
[method 4], and the desired compound (107 mg, 0.19 mmol) was
obtained as a mixture of diastereomers (yield 13.7%).
[0347] LC/MS [method 2]: R.sub.t=3.02 min; MS [ESIpos]: m/z=561
(M+H).sup.+
[0348] .sup.1H NMR (400 MHz, DMSO-d.sub.6): .delta. [ppm] 1.38 (d,
3H), 3.85 (dd, 1H), 4.00 (dd, 1H), 4.23-4.36 (m, 1H), 4.57-4.67 (m,
1H), 5.00-5.12 (m, 2H), 6.90 (br. s, 1H), 7.42 (td, 1H), 7.57-7.79
(m, 6H).
[0349] The two diastereomers were separated by preparative chiral
HPLC [sample preparation: 104 mg dissolved in 5 ml
ethanol/isohexane (1:1); injection volume: 0.5 ml; column: Daicel
Chiralcel.RTM. OX-H 5 .mu.m, 250.times.20 mm; eluent:
isohexane/ethanol 70:30; flow rate: 20 ml/min; temperature:
40.degree. C.; UV detection: 220 nm]. After separation, 56 mg of
diastereomer 1 (Example 35), which eluted first, and 29 mg of
diastereomer 2 (Example 36), which eluted later, were isolated.
Example 35
2-{[1-(2-Chloro-4-fluorophenyl)-5-(1-hydroxyethyl)-1H-H-1,2,4-triazol-3-yl-
]methyl}-5-(4-chlorophenyl)-4-[(2S)-3,3,3-trifluoro-2-hydroxypropyl]-2,4-d-
ihydro-3H-1,2,4-triazol-3-one (diastereomer 1)
[0350] Analytical chiral HPLC: R.sub.t=1.36 min, d.e.=100% [column:
Daicel Chiralpack OX-3 3 .mu.m, 50.times.4.6 mm; eluent:
isohexane/ethanol 70:30; flow rate: 1 ml/min; UV detection: 220
nm].
[0351] .sup.1H NMR (400 MHz, DMSO-d.sub.6): .delta. [ppm] 1.38 (d,
3H), 3.85 (dd, 1H), 4.00 (dd, 1H), 4.23-4.36 (m, 1H), 4.61 (quin,
1H), 5.01-5.11 (m, 2H), 5.51 (d, 1H), 6.89 (d, 1H), 7.42 (td, 1H),
7.60-7.65 (m, 2H), 7.66-7.72 (m, 1H), 7.72-7.78 (m, 3H).
Example 36
2-{[1-(2-Chloro-4-fluorophenyl)-5-(1-hydroxyethyl)-1H-1,2,4-triazol-3-yl]m-
ethyl}-5-(4-chlorophenyl)-4-[(2S)-3,3,3-trifluoro-2-hydroxypropyl]-2,4-dih-
ydro-3H-1,2,4-triazol-3-one (diastereomer 2)
[0352] Analytical chiral HPLC: R.sub.t=1.72 min, d.e.=100% [column:
Daicel Chiralpack OX-3 3 .mu.m, 50.times.4.6 mm; eluent:
isohexane/ethanol 70:30; flow rate: 1 ml/min; UV detection: 220
nm].
[0353] .sup.1H NMR (400 MHz, DMSO-d.sub.6): .delta. [ppm] 1.38 (d,
3H), 3.85 (dd, 1H), 4.00 (dd, 1H), 4.23-4.37 (m, 1H), 4.61 (quin,
1H), 5.06 (s, 2H), 5.51 (d, 1H), 6.90 (d, 1H), 7.42 (td, 1H),
7.60-7.65 (m, 2H), 7.66-7.72 (m, 1H), 7.72-7.78 (m, 3H).
Example 37
5-(4-Chlorophenyl)-2-({1-(5-fluoro-2-methoxyphenyl)-5-[(1RS)-1-hydroxyethy-
l]-1H-1,2,4-triazol-3-yl}methyl)-4-[(2S)-3,3,3-trifluoro-2-hydroxypropyl]--
2,4-dihydro-3H-1,2,4-triazol-3-one (diastereomeric mixture)
##STR00037##
[0355] To a solution of
5-(4-chlorophenyl)-2-({5-[(1RS)-1-hydroxyethyl]-1H-1,2,4-triazol-3-yl}met-
hyl)-4-[(2S)-3,3,3-trifluoro-2-hydroxypropyl]-2,4-dihydro-3H-1,2,4-triazol-
-3-one (600 mg, 1.39 mmol) in pyridine (18 ml) were added
(5-fluoro-2-methoxyphenyl)boronic acid (471.22 mg, 2.77 mmol) and
copper(II) acetate (503.6 mg, 2.77 mmol). The reaction mixture was
heated to 60.degree. C. for 2 h and then stirred at room
temperature for three days. The resulting reaction mixture was
concentrated in vacuo, then diluted with ethyl acetate and quenched
with aqueous hydrochloric acid (0.5 M). After phase separation, the
aqueous phase was extracted twice with ethyl acetate. The combined
organic phases were dried over sodium sulfate, filtered, and
concentrated in vacuo. The crude product was purified by
preparative HPLC [method 4], and the desired compound (62.2 mg,
0.11 mmol) was obtained as a mixture of diastereomers (yield
8.1%).
[0356] LC/MS [method 2]: R.sub.t=2.93 min; MS [ESIpos]: m/z=557
(M+H).sup.+
[0357] .sup.1H NMR (400 MHz, DMSO-d.sub.6): .delta. [ppm] 1.37 (d,
3H), 3.75 (s, 3H), 3.80-3.89 (m, 1H), 4.00 (dd, 1H), 4.24-4.36 (m,
1H), 4.52-4.62 (m, 1H), 4.99-5.04 (m, 2H), 6.90 (t, 1H), 7.26 (dd,
1H), 7.33 (dd, 1H), 7.37-7.44 (m, 1H), 7.60-7.65 (m, 2H), 7.73-7.78
(m, 2H).
[0358] The two diastereomers were separated by preparative chiral
HPLC [sample preparation: 55.4 mg dissolved in 6 ml
ethanol/isohexane (1:1); injection volume: 2 ml; column: Daicel
Chiralcel.RTM. OX-H 5 .mu.m, 250.times.20 mm; eluent:
isohexane/ethanol 80:20; flow rate: 20 ml/min; temperature:
40.degree. C.; UV detection: 220 nm]. After separation, 23 mg of
diastereomer 1 (Example 38), which eluted first, and 21 mg of
diastereomer 2 (Example 39), which eluted later, were isolated.
Example 38
5-(4-Chlorophenyl)-2-{[1-(5-fluoro-2-methoxyphenyl)-5-(1-hydroxyethyl)-1H--
1,2,4-triazol-3-yl]-methyl}-4-[(2S)-3,3,3-trifluoro-2-hydroxypropyl]-2,4-d-
ihydro-3H-1,2,4-triazol-3-one (diastereomer 1)
[0359] LC/MS [method 2]: R.sub.t=2.91 min; MS [ESIpos]: m/z=557
(M+H).sup.+
[0360] Analytical chiral HPLC: R.sub.t=2.13 min, d.e.=100% [column:
Daicel Chiralpack OX-3 3 .mu.m, 50.times.4.6 mm; eluent:
isohexane/ethanol 80:20; flow rate: 1 ml/min; temperature:
30.degree. C.; UV detection: 220 nm].
[0361] .sup.1H NMR (400 MHz, DMSO-d.sub.6): .delta. [ppm] 1.37 (d,
3H), 3.75 (s, 3H), 3.81-3.89 (m, 1H), 4.00 (dd, 1H), 4.25-4.35 (m,
1H), 4.53-4.62 (m, 1H), 4.99-5.10 (m, 2H), 5.40 (d, 1H), 6.89 (d,
1H), 7.26 (dd, 1H), 7.33 (dd, 1H), 7.41 (td, 1H), 7.60-7.66 (m,
2H), 7.73-7.79 (m, 2H).
Example 39
5-(4-Chlorophenyl)-2-{[1-(5-fluoro-2-methoxyphenyl)-5-(1-hydroxyethyl)-1H--
1,2,4-triazol-3-yl]-methyl}-4-[(2S)-3,3,3-trifluoro-2-hydroxypropyl]-2,4-d-
ihydro-3H-1,2,4-triazol-3-one (diastereomer 2)
[0362] LC/MS [method 2]: R.sub.t=2.90 min; MS [ESIpos]: m/z=557
(M+H).sup.+
[0363] Analytical chiral HPLC: R.sub.t=2.75 min, d.e.=100% [column:
Daicel Chiralpack OX-3 3 .mu.m, 50.times.4.6 mm; eluent:
isohexane/ethanol 80:20; flow rate: 1 ml/min; temperature:
30.degree. C.; UV detection: 220 nm].
[0364] .sup.1H NMR (400 MHz, DMSO-d.sub.6): .delta. [ppm] 1.37 (d,
3H), 3.75 (s, 3H), 3.85 (dd, 1H), 4.00 (dd, 1H), 4.24-4.36 (m, 1H),
4.57 (quin, 1H), 4.99-5.10 (m, 2H), 5.40 (d, 1H), 6.91 (d, 1H),
7.26 (dd, 1H), 7.33 (dd, 1H), 7.41 (td, 1H), 7.60-7.65 (m, 2H),
7.73-7.78 (m, 2H).
Example 40
5-(4-Chlorophenyl)-2-({1-(2-fluorophenyl)-5-[(1RS)-1-hydroxyethyl]-1H-1,2,-
4-triazol-3-yl}-methyl)-4-[(2S)-3,3,3-trifluoro-2-hydroxypropyl]-2,4-dihyd-
ro-3H-1,2,4-triazol-3-one (diastereomeric mixture)
##STR00038##
[0366] To a solution of
5-(4-chlorophenyl)-2-({5-[(1RS)-1-hydroxyethyl]-1H-1,2,4-triazol-3-yl}met-
hyl)-4-[(2S)-3,3,3-trifluoro-2-hydroxypropyl]-2,4-dihydro-3H-1,2,4-triazol-
-3-one (600 mg, 1.39 mmol) in pyridine (18 ml) were added
(2-fluorophenyl)boronic acid (387.96 mg, 2.77 mmol) and copper(II)
acetate (503.6 mg, 2.77 mmol). The reaction mixture was heated to
60.degree. C. for 2 h and then stirred at room temperature for 3
days. Over this time, two additional portions of boronic acid
(387.96 mg in total, 2.77 mmol) were added. After this, the
resulting reaction mixture was concentrated in vacuo, then diluted
with ethyl acetate and quenched with aqueous hydrochloric acid (0.5
M). After phase separation, the aqueous phase was extracted twice
with ethyl acetate. The combined organic phases were dried over
sodium sulfate, filtered, and concentrated in vacuo. The crude
product was purified by preparative HPLC [method 4], and the
desired compound (30.1 mg, 0.06 mmol) was obtained as a mixture of
diastereomers (yield 4.1%).
[0367] LC/MS [method 2]: R.sub.t=2.84 min; MS [ESIpos]: m/z=527
(M+H).sup.+
[0368] .sup.1H NMR (400 MHz, DMSO-d.sub.6): .delta. [ppm] 1.40 (d,
3H), 3.85 (dd, 1H), 4.00 (dd, 1H), 4.29 (br. s, 1H), 4.69 (q, 1H),
5.01-5.12 (m, 2H), 6.89 (br. s, 1H), 7.38 (t, 1H), 7.48 (t, 1H),
7.57-7.66 (m, 4H), 7.71-7.80 (m, 2H).
[0369] The two diastereomers were separated by preparative chiral
HPLC [sample preparation: 26 mg dissolved in 4 ml ethanol/isohexane
(1:1); injection volume: 2 ml; column: Daicel Chiralcel.RTM. OX-H 5
.mu.m, 250.times.20 mm; eluent: isohexane/ethanol 80:20; flow rate:
25 ml/min; temperature: 40.degree. C.; UV detection: 220 nm]. After
separation, 11 mg of diastereomer 1 (Example 41), which eluted
first, and 9 mg of diastereomer 2 (Example 42), which eluted later,
were isolated.
Example 41
5-(4-Chlorophenyl)-2-{[1-(2-fluorophenyl)-5-(1-hydroxyethyl)-1H-1,2,4-tria-
zol-3-yl]methyl}-4-[(2S)-3,3,3-trifluoro-2-hydroxypropyl]-2,4-dihydro-3H-1-
,2,4-triazol-3-one (diastereomer 1)
[0370] LC/MS [method 2]: R.sub.t=2.83 min; MS [ESIpos]: m/z=527
(M+H).sup.+
[0371] Analytical chiral HPLC: R.sub.t=2.32 min, d.e.=100% [column:
Daicel Chiralpack OX-3 3 .mu.m, 50.times.4.6 mm; eluent:
isohexane/ethanol 80:20; flow rate: 1 ml/min; temperature:
30.degree. C.; UV detection: 220 nm].
[0372] .sup.1H NMR (400 MHz, DMSO-d.sub.6): .delta. [ppm] 1.40 (d,
3H), 3.85 (dd, 1H), 4.00 (dd, 1H), 4.24-4.36 (m, 1H), 4.69 (quin,
1H), 5.01-5.12 (m, 2H), 5.53 (d, 1H), 6.89 (d, 1H), 7.38 (t, 1H),
7.48 (t, 1H), 7.57-7.66 (m, 4H), 7.72-7.78 (m, 2H).
Example 42
5-(4-Chlorophenyl)-2-{[1-(2-fluorophenyl)-5-(1-hydroxyethyl)-1H-1,2,4-tria-
zol-3-yl]methyl}-4-[(2S)-3,3,3-trifluoro-2-hydroxypropyl]-2,4-dihydro-3H-1-
,2,4-triazol-3-one (diastereomer 2)
[0373] LC/MS [method 2]: R.sub.t=2.82 min; MS [ESIpos]: m/z=527
(M+H).sup.+
[0374] Analytical chiral HPLC: R.sub.t=3.23 min, d.e.=100% [column:
Daicel Chiralpack OX-3 3 .mu.m, 50.times.4.6 mm; eluent:
isohexane/ethanol 80:20; flow rate: 1 ml/min; temperature:
30.degree. C.; UV detection: 220 nm].
[0375] .sup.1H NMR (400 MHz, DMSO-d.sub.6): .delta. [ppm] 1.40 (d,
3H), 3.84 (dd, 1H), 4.00 (dd, 1H), 4.24-4.35 (m, 1H), 4.69 (quin,
1H), 5.07 (s, 2H), 5.53 (d, 1H), 6.90 (d, 1H), 7.38 (t, 1H), 7.48
(t, 1H), 7.57-7.67 (m, 4H), 7.72-7.79 (m, 2H).
Example 43
2-({1-(3-Chloro-4-fluorophenyl)-5-[(1RS)-1-hydroxyethyl]-1H-1,2,4-triazol--
3-yl}methyl)-5-(4-chlorophenyl)-4-[(2S)-3,3,3-trifluoro-2-hydroxypropyl]-2-
,4-dihydro-3H-1,2,4-triazol-3-one (diastereomeric mixture)
##STR00039##
[0377] To a solution of
5-(4-chlorophenyl)-2-({5-[(1RS)-1-hydroxyethyl]-1H-1,2,4-triazol-3-yl}met-
hyl)-4-[(2S)-3,3,3-trifluoro-2-hydroxypropyl]-2,4-dihydro-3H-1,2,4-triazol-
-3-one (400 mg, 0.92 mmol) in pyridine (12 ml) were added
(3-chloro-4-fluorophenyl)boronic acid (322.6 mg, 1.85 mmol) and
copper(II) acetate (335.7 mg, 1.85 mmol). The reaction mixture was
heated to 60.degree. C. for 2 h and then stirred at room
temperature for 6 days. The resulting reaction mixture was
concentrated in vacuo, then diluted with ethyl acetate and quenched
with aqueous hydrochloric acid (0.5 M). After phase separation, the
aqueous phase was extracted twice with ethyl acetate. The combined
organic phases were dried over sodium sulfate, filtered, and
concentrated in vacuo. The crude product was purified by
preparative HPLC [method 4], and the desired compound (99.1 mg,
0.18 mmol) was obtained as a mixture of diastereomers (yield
19.1%).
[0378] LC/MS [method 3]: R.sub.t=1.31 min; MS [ESIpos]: m/z=561
(M+H).sup.+
[0379] .sup.1H NMR (400 MHz, DMSO-d.sub.6): .delta. [ppm] 1.46 (d,
3H), 3.85 (dd, 1H), 4.01 (dd, 1H), 4.30 (br. s, 1H), 4.80 (q, 1H),
5.01-5.13 (m, 2H), 6.90 (br. s, 1H), 7.59-7.70 (m, 4H), 7.72-7.78
(m, 2H), 7.93 (dd, 1H).
[0380] The two diastereomers were separated by preparative chiral
HPLC [sample preparation: 97.1 mg dissolved in 3 ml ethanol;
injection volume: 0.3 ml; column: Daicel Chiralcel.RTM. OX-H 5
.mu.m, 250.times.20 mm; eluent: isohexane/ethanol 80:20; flow rate:
15 ml/min; temperature: 25.degree. C.; UV detection: 220 nm]. After
separation, 40 mg of diastereomer 1 (Example 44), which eluted
first, and 42 mg of diastereomer 2 (Example 45), which eluted
later, were isolated.
Example 44
2-{[1-(3-Chloro-4-fluorophenyl)-5-(1-hydroxyethyl)-1H-H-1,2,4-triazol-3-yl-
]methyl}-5-(4-chlorophenyl)-4-[(2S)-3,3,3-trifluoro-2-hydroxypropyl]-2,4-d-
ihydro-3H-1,2,4-triazol-3-one (diastereomer 1)
[0381] LC/MS [method 2]: R.sub.t=3.22 min; MS [ESIpos]: m/z=561
(M+H).sup.+
[0382] Preparative chiral HPLC: R.sub.t=9.97 min, d.e.=100%
[column: Daicel Chiralcel.RTM. OX-H 5 .mu.m, 250.times.20 mm;
eluent: isohexane/ethanol 80:20; flow rate: 15 ml/min; temperature:
25.degree. C.; UV detection: 220 nm].
[0383] .sup.1H NMR (400 MHz, DMSO-d.sub.6): .delta. [ppm] 1.46 (d,
3H), 3.85 (dd, 1H), 4.01 (dd, 1H), 4.22-4.36 (m, 1H), 4.80 (quin,
1H), 5.01-5.12 (m, 2H), 5.75 (d, 1H), 6.89 (d, 1H), 7.58-7.70 (m,
4H), 7.71-7.78 (m, 2H), 7.93 (dd, 1H).
Example 45
2-{[1-(3-Chloro-4-fluorophenyl)-5-(1-hydroxyethyl)-1H-H-1,2,4-triazol-3-yl-
]methyl}-5-(4-chlorophenyl)-4-[(2S)-3,3,3-trifluoro-2-hydroxypropyl]-2,4-d-
ihydro-3H-1,2,4-triazol-3-one (diastereomer 2)
[0384] LC/MS [method 2]: R.sub.t=3.22 min; MS [ESIpos]: m/z=561
(M+H).sup.+
[0385] Preparative chiral HPLC: R.sub.t=11.35 min, d.e.=100%
[column: Daicel Chiralcel.RTM. OX-H 5 .mu.m, 250.times.20 mm;
eluent: isohexane/ethanol 80:20; flow rate: 15 ml/min; temperature:
25.degree. C.; UV detection: 220 nm].
[0386] .sup.1H NMR (400 MHz, DMSO-d.sub.6): .delta. [ppm] 1.46 (d,
3H), 3.85 (dd, 1H), 4.00 (dd, 1H), 4.25-4.35 (m, 1H), 4.81 (quin,
1H), 5.06 (s, 2H), 5.74 (d, 1H), 6.90 (d, 1H), 7.59-7.70 (m, 4H),
7.73-7.78 (m, 2H), 7.93 (dd, 1H).
Example 46
5-(4-Chlorophenyl)-2-({1-(3,5-dichlorophenyl)-5-[(1RS)-1-hydroxyethyl]-1H--
1,2,4-triazol-3-yl}-methyl)-4-[(2S)-3,3,3-trifluoro-2-hydroxypropyl]-2,4-d-
ihydro-3H-1,2,4-triazol-3-one (diastereomeric mixture)
##STR00040##
[0388] To a solution of
5-(4-chlorophenyl)-2-({5-[(1RS)-1-hydroxyethyl]-1H-1,2,4-triazol-3-yl}met-
hyl)-4-[(2S)-3,3,3-trifluoro-2-hydroxypropyl]-2,4-dihydro-3H-1,2,4-triazol-
-3-one (400 mg, 0.92 mmol) in pyridine (12 ml) were added
(3,5-dichlorophenyl)boronic acid (352.73 mg, 1.85 mmol) and
copper(II) acetate (335.7 mg, 1.85 mmol). The reaction mixture was
heated to 60.degree. C. for 2 h and then stirred at room
temperature for 6 days. The resulting reaction mixture was
concentrated in vacuo, then diluted with ethyl acetate and quenched
with aqueous hydrochloric acid (0.5 M). After phase separation, the
aqueous phase was extracted twice with ethyl acetate. The combined
organic phases were dried over sodium sulfate, filtered, and
concentrated in vacuo. The crude product was purified by
preparative HPLC [method 4], and the desired compound (105.5 mg,
0.18 mmol) was obtained as a mixture of diastereomers (yield
19.8%).
[0389] LC/MS [method 3]: R.sub.t=1.39 min; MS [ESIpos]: m/z=577
(M+H).sup.+
[0390] .sup.1H NMR (400 MHz, DMSO-d.sub.6): .delta. [ppm] 1.48 (d,
3H), 3.85 (dd, 1H), 4.01 (dd, 1H), 4.23-4.36 (m, 1H), 4.82-4.90 (m,
1H), 5.02-5.12 (m, 2H), 6.84-6.94 (m, 1H), 7.59-7.65 (m, 2H),
7.72-7.82 (m, 5H).
[0391] The two diastereomers were separated by preparative chiral
HPLC [sample preparation: 103.5 mg dissolved in 14 ml
ethanol/isohexane (1:1); injection volume: 2 ml; column: Daicel
Chiralcel.RTM. OX-H 5 .mu.m, 250.times.20 mm; eluent:
isohexane/ethanol 80:20; flow rate: 20 ml/min; temperature:
30.degree. C.; UV detection: 220 nm]. After separation, 29.2 mg of
diastereomer 1 (Example 47), which eluted first, and 28.9 mg of
diastereomer 2 (Example 48), which eluted later, were isolated.
Example 47
5-(4-Chlorophenyl)-2-{[1-(3,5-dichlorophenyl)-5-(1-hydroxyethyl)-1H-1,2,4--
triazol-3-yl]methyl}-4-[(2S)-3,3,3-trifluoro-2-hydroxypropyl]-2,4-dihydro--
3H-1,2,4-triazol-3-one (diastereomer 1)
[0392] Analytical chiral HPLC: R.sub.t=1.49 min, d.e.=100% [column:
Daicel Chiralpack OX-3 3 .mu.m, 50.times.4.6 mm; eluent:
isohexane/ethanol 80:20; flow rate: 1 ml/min; temperature:
30.degree. C.; UV detection: 220 nm].
[0393] .sup.1H NMR (400 MHz, DMSO-d.sub.6): .delta. [ppm] 1.48 (d,
3H), 3.85 (dd, 1H), 4.01 (dd, 1H), 4.24-4.35 (m, 1H), 4.86 (quin,
1H), 5.01-5.12 (m, 2H), 5.82 (d, 1H), 6.88 (d, 1H), 7.58-7.66 (m,
2H), 7.72-7.82 (m, 5H).
Example 48
5-(4-Chlorophenyl)-2-{[1-(3,5-dichlorophenyl)-5-(1-hydroxyethyl)-1H-1,2,4--
triazol-3-yl]methyl}-4-[(2S)-3,3,3-trifluoro-2-hydroxypropyl]-2,4-dihydro--
3H-1,2,4-triazol-3-one (diastereomer 2)
[0394] Analytical chiral HPLC: R.sub.t=2.02 min, d.e.=99.8%
[column: Daicel Chiralpack OX-3 3 .mu.m, 50.times.4.6 mm; eluent:
isohexane/ethanol 80:20; flow rate: 1 ml/min; temperature:
30.degree. C.; UV detection: 220 nm].
[0395] .sup.1H NMR (400 MHz, DMSO-d.sub.6): .delta. [ppm] 1.48 (d,
3H), 3.85 (dd, 1H), 4.01 (dd, 1H), 4.23-4.36 (m, 1H), 4.86 (quin,
1H), 5.07 (s, 2H), 5.82 (d, 1H), 6.89 (d, 1H), 7.59-7.65 (m, 2H),
7.72-7.81 (m, 5H).
Example 49
5-(4-Chlorophenyl)-2-({1-(2,5-dichlorophenyl)-5-[(1RS)-1-hydroxyethyl]-1H--
1,2,4-triazol-3-yl}-methyl)-4-[(2S)-3,3,3-trifluoro-2-hydroxypropyl]-2,4-d-
ihydro-3H-1,2,4-triazol-3-one (diastereomeric mixture)
##STR00041##
[0397] To a solution of
5-(4-chlorophenyl)-2-({5-[(1RS)-1-hydroxyethyl]-1H-1,2,4-triazol-3-yl}met-
hyl)-4-[(2S)-3,3,3-trifluoro-2-hydroxypropyl]-2,4-dihydro-3H-1,2,4-triazol-
-3-one (400 mg, 0.92 mmol) in pyridine (12 ml) were added
(2,5-dichlorophenyl)boronic acid (352.73 mg, 1.85 mmol) and
copper(II) acetate (335.75 mg, 1.85 mmol). The reaction mixture was
heated to 60.degree. C. for 2 h and then stirred at room
temperature for three days, after which extra boronic acid (100 mg,
0.52 mmol) was added due to incomplete conversion. The reaction
mixture was further stirred at room temperature for 6 days. Over
this time, another portion of boronic acid (100 mg, 0.52 mmol) was
added. After this, the resulting reaction mixture was concentrated
in vacuo, then diluted with ethyl acetate and quenched with aqueous
hydrochloric acid (0.5 M). After phase separation, the aqueous
phase was extracted twice with ethyl acetate. The combined organic
phases were dried over sodium sulfate, filtered, and concentrated
in vacuo. The crude product was purified by preparative HPLC
[method 4], and the desired compound (52.8 mg, 0.09 mmol, 97%
purity) was obtained as a mixture of diastereomers (yield
9.6%).
[0398] LC/MS [method 3]: R.sub.t=1.33 min; MS [ESIpos]: m/z=577
(M+H).sup.+
[0399] .sup.1H NMR (400 MHz, DMSO-d.sub.6): .delta. [ppm] 1.40 (d,
3H), 3.85 (dd, 1H), 4.00 (dd, 1H), 4.23-4.35 (m, 1H), 4.63-4.72 (m,
1H), 5.01-5.12 (m, 2H), 6.89 (br. s, 1H), 7.60-7.65 (m, 2H),
7.67-7.78 (m, 4H), 7.81 (br. d, 1H).
[0400] The two diastereomers were separated by preparative chiral
HPLC (SFC) [sample preparation: 50 mg dissolved in 10 ml methanol;
injection volume: 0.5 ml; column: Daicel Chiralcel.RTM. OX-H 5
.mu.m, 250.times.20 mm; eluent: carbon dioxide/methanol 82:18; flow
rate: 80 ml/min; temperature: 40.degree. C.; UV detection: 210 nm].
After separation, 20.3 mg of diastereomer 1 (Example 50), which
eluted first, and 24.1 mg of diastereomer 2 (Example 51), which
eluted later, were isolated.
Example 50
5-(4-Chlorophenyl)-2-{[1-(2,5-dichlorophenyl)-5-(1-hydroxyethyl)-1H-1,2,4--
triazol-3-yl]methyl}-4-[(2S)-3,3,3-trifluoro-2-hydroxypropyl]-2,4-dihydro--
3H-1,2,4-triazol-3-one (diastereomer 1)
[0401] LC/MS [method 3]: R.sub.t=1.30 min; MS [ESIpos]: m/z=577
(M+H).sup.+
[0402] Analytical chiral HPLC (SFC): R.sub.t=2.87 min, d.e.=100%
[column: Daicel Chiralcel.RTM. OX-3, 250.times.4 mm; eluent: carbon
dioxide/methanol (5%.fwdarw.60%); flow rate: 3 ml/min; UV
detection: 220 nm].
[0403] .sup.1H NMR (400 MHz, DMSO-d.sub.6): .delta. [ppm] 1.40 (d,
3H), 3.85 (dd, 1H), 4.00 (dd, 1H), 4.23-4.36 (m, 1H), 4.67 (quin,
1H), 5.01-5.12 (m, 2H), 5.53 (d, 1H), 6.89 (d, 1H), 7.60-7.65 (m,
2H), 7.67-7.78 (m, 4H), 7.81 (br. d, 1H).
Example 51
5-(4-Chlorophenyl)-2-{[1-(2,5-dichlorophenyl)-5-(1-hydroxyethyl)-1H-1,2,4--
triazol-3-yl]methyl}-4-[(2S)-3,3,3-trifluoro-2-hydroxypropyl]-2,4-dihydro--
3H-1,2,4-triazol-3-one (diastereomer 2)
[0404] LC/MS [method 3]: R.sub.t=1.30 min; MS [ESIpos]: m/z=577
(M+H).sup.+
[0405] Analytical chiral HPLC (SFC): R.sub.t=3.11 min, d.e.=100%
[column: Daicel Chiralcel.RTM. OX-3, 250.times.4 mm; eluent: carbon
dioxide/methanol (5%.fwdarw.60%); flow rate: 3 ml/min; UV
detection: 220 nm].
[0406] .sup.1H NMR (400 MHz, DMSO-d.sub.6): .delta. [ppm] 1.40 (d,
3H), 3.85 (dd, 1H), 4.00 (dd, 1H), 4.23-4.35 (m, 1H), 4.67 (quin,
1H), 5.01-5.12 (m, 2H), 5.53 (d, 1H), 6.89 (d, 1H), 7.59-7.65 (m,
2H), 7.66-7.78 (m, 4H), 7.81 (br. d, 1H).
Example 52
2-({1-(3-Chloro-2-fluorophenyl)-5-[(1RS)-1-hydroxyethyl]-1H-1,2,4-triazol--
3-yl}methyl)-5-(4-chlorophenyl)-4-[(2S)-3,3,3-trifluoro-2-hydroxypropyl]-2-
,4-dihydro-3H-1,2,4-triazol-3-one (diastereomeric mixture)
##STR00042##
[0408] To a solution of
5-(4-chlorophenyl)-2-({5-[(1RS)-1-hydroxyethyl]-1H-1,2,4-triazol-3-yl}met-
hyl)-4-[(2S)-3,3,3-trifluoro-2-hydroxypropyl]-2,4-dihydro-3H-1,2,4-triazol-
-3-one (400 mg, 0.92 mmol) in pyridine (12 ml) were added
(3-chloro-2-fluorophenyl)boronic acid (322.31 mg, 1.85 mmol) and
copper(II) acetate (335.75 mg, 1.85 mmol). The reaction mixture was
heated to 60.degree. C. for 2 h and then stirred at room
temperature for 12 days. Over this time, extra boronic acid (322.31
mg in total, 1.85 mmol) was added portionwise in a daily fashion.
After this, the resulting reaction mixture was concentrated in
vacuo, then diluted with ethyl acetate and quenched with aqueous
hydrochloric acid (0.5 M). After phase separation, the aqueous
phase was extracted twice with ethyl acetate. The combined organic
phases were dried over sodium sulfate, filtered, and concentrated
in vacuo. The crude product was purified by preparative HPLC
[method 4], and the desired compound (15.9 mg, 0.03 mmol, 97%
purity) was obtained as a mixture of diastereomers (yield 3%).
[0409] LC/MS [method 2]: R.sub.t=3.12 min; MS [ESIpos]: m/z=561
(M+H).sup.+
[0410] .sup.1H NMR (500 MHz, DMSO-d.sub.6): .delta. [ppm] 1.42 (d,
3H), 3.85 (dd, 1H), 4.00 (dd, 1H), 4.23-4.35 (m, 1H), 4.75 (q, 1H),
5.02-5.12 (m, 2H), 5.57 (br. s, 1H), 6.89 (br. d, 1H), 7.40 (td,
1H), 7.60-7.65 (m, 3H), 7.72-7.77 (m, 2H), 7.81 (ddd, 1H).
[0411] The two diastereomers were separated by preparative chiral
HPLC [sample preparation: 14 mg dissolved in 1 ml ethanol/isohexane
(1:1); injection volume: 1 ml; column: Daicel Chiralcel.RTM. OX-H 5
.mu.m, 250.times.20 mm; eluent: isohexane/ethanol 80:20; flow rate:
15 ml/min; temperature: 25.degree. C.; UV detection: 220 nm]. After
separation, 6 mg of diastereomer 1 (Example 53), which eluted
first, and 6 mg of diastereomer 2 (Example 54), which eluted later,
were isolated.
Example 53
2-{[1-(3-Chloro-2-fluorophenyl)-5-(1-hydroxyethyl)-1H-H-1,2,4-triazol-3-yl-
]methyl}-5-(4-chlorophenyl)-4-[(2S)-3,3,3-trifluoro-2-hydroxypropyl]-2,4-d-
ihydro-3H-1,2,4-triazol-3-one (diastereomer 1)
[0412] LC/MS [method 2]: R.sub.t=3.14 min; MS [ESIpos]: m/z=561
(M+H).sup.+
[0413] Analytical chiral HPLC: R.sub.t=5.49 min, d.e.=100% [column:
Daicel Chiralcel.RTM. OX-H 5 .mu.m, 250.times.4.6 mm; eluent:
isohexane/ethanol 70:30+0.2% TFA and 1% water; flow rate: 1 ml/min;
temperature: 40.degree. C.; UV detection: 220 nm].
[0414] .sup.1H NMR (500 MHz, DMSO-d.sub.6): .delta. [ppm] 1.42 (d,
3H), 3.85 (dd, 1H), 4.00 (dd, 1H), 4.24-4.34 (m, 1H), 4.75 (quin,
1H), 5.03-5.11 (m, 2H), 5.57 (d, 1H), 6.89 (d, 1H), 7.40 (td, 1H),
7.60-7.65 (m, 3H), 7.73-7.77 (m, 2H), 7.81 (ddd, 1H).
Example 54
2-{[1-(3-Chloro-2-fluorophenyl)-5-(1-hydroxyethyl)-1H-H-1,2,4-triazol-3-yl-
]methyl}-5-(4-chlorophenyl)-4-[(2S)-3,3,3-trifluoro-2-hydroxypropyl]-2,4-d-
ihydro-3H-1,2,4-triazol-3-one (diastereomer 2)
[0415] LC/MS [method 2]: R.sub.t=3.13 min; MS [ESIpos]: m/z=561
(M+H).sup.+
[0416] Analytical chiral HPLC: R.sub.t=6.16 min, d.e.=100% [column:
Daicel Chiralcel.RTM. OX-H 5 .mu.m, 250.times.4.6 mm; eluent:
isohexane/ethanol 70:30+0.2% TFA and 1% water; flow rate: 1 ml/min;
temperature: 40.degree. C.; UV detection: 220 nm].
[0417] .sup.1H NMR (500 MHz, DMSO-d.sub.6): .delta. [ppm] 1.42 (d,
3H), 3.85 (dd, 1H), 4.00 (dd, 1H), 4.25-4.34 (m, 1H), 4.75 (quin,
1H), 5.07 (s, 2H), 5.57 (d, 1H), 6.89 (d, 1H), 7.40 (td, 1H),
7.60-7.65 (m, 3H), 7.73-7.77 (m, 2H), 7.81 (ddd, 1H).
Example 55
5-(4-Chlorophenyl)-2-({1-[3-(difluoromethoxy)phenyl]-5-[(1RS)-1-hydroxyeth-
yl]-1H-1,2,4-triazol-3-yl}methyl)-4-[(2S)-3,3,3-trifluoro-2-hydroxypropyl]-
-2,4-dihydro-3H-1,2,4-triazol-3-one (diastereomeric mixture)
##STR00043##
[0419] To a solution of
5-(4-chlorophenyl)-2-({5-[(1RS)-1-hydroxyethyl]-1H-1,2,4-triazol-3-yl}met-
hyl)-4-[(2S)-3,3,3-trifluoro-2-hydroxypropyl]-2,4-dihydro-3H-1,2,4-triazol-
-3-one (400 mg, 0.92 mmol) in pyridine (12 ml) were added
[3-(difluoromethoxy)phenyl]boronic acid (347.40 mg, 1.85 mmol) and
copper(II) acetate (335.75 mg, 1.85 mmol). The reaction mixture was
heated to 60.degree. C. for 2 h and then stirred at room
temperature for 6 days, after which extra boronic acid (100 mg,
0.53 mmol) was added due to incomplete conversion. The reaction
mixture was stirred at room temperature for two additional days.
The resulting reaction mixture was concentrated in vacuo, then
diluted with ethyl acetate and quenched with aqueous hydrochloric
acid (0.5 M). After phase separation, the aqueous phase was
extracted twice with ethyl acetate. The combined organic phases
were dried over sodium sulfate, filtered, and concentrated in
vacuo. The crude product was purified by preparative HPLC [method
4], and the desired compound (60.3 mg, 0.10 mmol) was obtained as a
mixture of diastereomers (yield 11.4%).
[0420] LC/MS [method 3]: R.sub.t=1.28 min; MS [ESIpos]: m/z=575
(M+H).sup.+
[0421] .sup.1H NMR (400 MHz, DMSO-d.sub.6): .delta. [ppm] 1.47 (d,
3H), 3.85 (dd, 1H), 4.01 (dd, 1H), 4.25-4.35 (m, 1H), 4.78-4.85 (m,
1H), 5.03-5.12 (m, 2H), 6.89 (br. s, 1H), 7.33 (t, 1H), 7.31-7.35
(m, 1H), 7.48-7.56 (m, 2H), 7.59-7.65 (m, 3H), 7.72-7.78 (m,
2H).
[0422] The two diastereomers were separated by preparative chiral
HPLC [sample preparation: 58 mg dissolved in 2 ml ethanol;
injection volume: 0.7 ml; column: Daicel Chiralcel.RTM. OX-H 5
.mu.m, 250.times.20 mm; eluent: isohexane/ethanol 80:20; flow rate:
15 ml/min; temperature: 35.degree. C.; UV detection: 220 nm]. After
separation, 20.7 mg of diastereomer 1 (Example 56), which eluted
first, and 17.7 mg of diastereomer 2 (Example 57), which eluted
later, were isolated.
Example 56
5-(4-Chlorophenyl)-2-({1-[3-(difluoromethoxy)phenyl]-5-(1-hydroxyethyl)-1H-
-1,2,4-triazol-3-yl}-methyl)-4-[(2S)-3,3,3-trifluoro-2-hydroxypropyl]-2,4--
dihydro-3H-1,2,4-triazol-3-one (diastereomer 1)
[0423] Analytical chiral HPLC: R.sub.t=5.57 min, d.e.=98.7%
[column: Daicel Chiralcel.RTM. OX-H 5, 250.times.4.6 mm; eluent:
isohexane/ethanol 70:30+0.2% TFA and 1% water; flow rate: 1 ml/min;
temperature: 35.degree. C.; UV detection: 220 nm].
[0424] .sup.1H NMR (400 MHz, DMSO-d.sub.6): .delta. [ppm] 1.47 (d,
3H), 3.85 (dd, 1H), 4.01 (dd, 1H), 4.30 (br. s, 1H), 4.81 (q, 1H),
5.02-5.13 (m, 2H), 6.88 (br. s, 1H), 7.33 (t, 1H), 7.30-7.35 (m,
1H), 7.48-7.56 (m, 2H), 7.59-7.65 (m, 3H), 7.72-7.78 (m, 2H).
Example 57
5-(4-Chlorophenyl)-2-({1-[3-(difluoromethoxy)phenyl]-5-(1-hydroxyethyl)-1H-
-1,2,4-triazol-3-yl}-methyl)-4-[(2S)-3,3,3-trifluoro-2-hydroxypropyl]-2,4--
dihydro-3H-1,2,4-triazol-3-one (diastereomer 2)
[0425] Analytical chiral HPLC: R.sub.t=6.70 min, d.e.=100% [column:
Daicel Chiralcel.RTM. OX-H 5, 250.times.4.6 mm; eluent:
isohexane/ethanol 70:30+0.2% TFA and 1% water; flow rate: 1 ml/min;
temperature: 35.degree. C.; UV detection: 220 nm].
[0426] .sup.1H NMR (400 MHz, DMSO-d.sub.6): .delta. [ppm] 1.47 (d,
3H), 3.85 (dd, 1H), 4.01 (dd, 1H), 4.23-4.35 (m, 1H), 4.82 (quin,
1H), 5.07 (s, 2H), 5.75 (d, 1H), 6.90 (d, 1H), 7.33 (t, 1H),
7.30-7.35 (m, 1H), 7.48-7.56 (m, 2H), 7.59-7.65 (m, 3H), 7.72-7.78
(m, 2H).
Example 58
2-({1-(2-Chloro-5-fluorophenyl)-5-[(1RS)-1-hydroxyethyl]-1H-1,2,4-triazol--
3-yl}methyl)-5-(4-chlorophenyl)-4-[(2S)-3,3,3-trifluoro-2-hydroxypropyl]-2-
,4-dihydro-3H-1,2,4-triazol-3-one (diastereomeric mixture)
##STR00044##
[0428] To a solution of
5-(4-chlorophenyl)-2-({5-[(1RS)-1-hydroxyethyl]-1H-1,2,4-triazol-3-yl}met-
hyl)-4-[(2S)-3,3,3-trifluoro-2-hydroxypropyl]-2,4-dihydro-3H-1,2,4-triazol-
-3-one (400 mg, 0.92 mmol) in pyridine (12 ml) were added
(2-chloro-5-fluorophenyl)boronic acid (322.31 mg, 1.85 mmol) and
copper(II) acetate (335.75 mg, 1.85 mmol). The reaction mixture was
heated to 60.degree. C. for 2 h and then stirred at room
temperature for 10 days. Over this time, extra boronic acid (322.31
mg in total, 1.85 mmol) was added portionwise in a daily fashion.
The resulting reaction mixture was concentrated in vacuo, then
diluted with ethyl acetate and quenched with aqueous hydrochloric
acid (0.5 M). After phase separation, the aqueous phase was
extracted twice with ethyl acetate. The combined organic phases
were dried over sodium sulfate, filtered, and concentrated in
vacuo. The crude product was purified by preparative HPLC [method
4], and the desired compound (61 mg, 0.11 mmol, 98% purity) was
obtained as a mixture of diastereomers (yield 11.5%).
[0429] LC/MS [method 2]: R.sub.t=3.02 min; MS [ESIpos]: m/z=561
(M+H).sup.+
[0430] .sup.1H NMR (400 MHz, DMSO-d.sub.6): .delta. [ppm] 1.40 (d,
3H), 3.85 (dd, 1H), 4.00 (dd, 1H), 4.24-4.36 (m, 1H), 4.63-4.72 (m,
1H), 5.01-5.13 (m, 2H), 5.52 (br. s, 1H), 6.90 (dd, 1H), 7.52 (td,
1H), 7.60-7.69 (m, 3H), 7.73-7.79 (m, 3H).
[0431] The two diastereomers were separated by preparative chiral
HPLC [sample preparation: 58 mg dissolved in 3 ml ethanol/isohexane
(2:1); injection volume: 1 ml; column: Daicel Chiralcel.RTM. OX-H
m, 250.times.20 mm; eluent: isohexane/ethanol 80:20; flow rate: 15
ml/min; temperature: 25.degree. C.; UV detection: 220 nm]. After
separation, 25 mg of diastereomer 1 (Example 59), which eluted
first, and 25 mg of diastereomer 2 (Example 60), which eluted
later, were isolated.
Example 59
2-{[1-(2-Chloro-5-fluorophenyl)-5-(1-hydroxyethyl)-1H-H-1,2,4-triazol-3-yl-
]methyl}-5-(4-chlorophenyl)-4-[(2S)-3,3,3-trifluoro-2-hydroxypropyl]-2,4-d-
ihydro-3H-1,2,4-triazol-3-one (diastereomer 1)
[0432] LC/MS [method 2]: R.sub.t=3.02 min; MS [ESIpos]: m/z=561
(M+H).sup.+
[0433] Analytical chiral HPLC: R.sub.t=5.43 min, d.e.=100% [column:
Daicel Chiralcel.RTM. OX-H 5 .mu.m, 250.times.4.6 mm; eluent:
isohexane/ethanol 70:30+0.2% TFA and 1% water; flow rate: 1 ml/min;
temperature: 40.degree. C.; UV detection: 220 nm].
[0434] .sup.1H NMR (400 MHz, DMSO-d.sub.6): .delta. [ppm] 1.40 (d,
3H), 3.85 (dd, 1H), 4.00 (dd, 1H), 4.23-4.35 (m, 1H), 4.67 (quin,
1H), 5.01-5.12 (m, 2H), 5.53 (d, 1H), 6.89 (d, 1H), 7.52 (td, 1H),
7.60-7.68 (m, 3H), 7.72-7.79 (m, 3H).
Example 60
2-{[1-(2-Chloro-5-fluorophenyl)-5-(1-hydroxyethyl)-1H-H-1,2,4-triazol-3-yl-
]methyl}-5-(4-chlorophenyl)-4-[(2S)-3,3,3-trifluoro-2-hydroxypropyl]-2,4-d-
ihydro-3H-1,2,4-triazol-3-one (diastereomer 2)
[0435] LC/MS [method 2]: R.sub.t=3.02 min; MS [ESIpos]: m/z=561
(M+H).sup.+
[0436] Analytical chiral HPLC: R.sub.t=6.11 min, d.e.=100% [column:
Daicel Chiralcel.RTM. OX-H 5 .mu.m, 250.times.4.6 mm; eluent:
isohexane/ethanol 70:30+0.2% TFA and 1% water; flow rate: 1 ml/min;
temperature: 40.degree. C.; UV detection: 220 nm].
[0437] .sup.1H NMR (400 MHz, DMSO-d.sub.6): .delta. [ppm] 1.40 (d,
3H), 3.85 (dd, 1H), 4.00 (dd, 1H), 4.30 (br. s, 1H), 4.67 (quin,
1H), 5.07 (s, 2H), 5.53 (d, 1H), 6.90 (d, 1H), 7.52 (td, 1H),
7.60-7.68 (m, 3H), 7.73-7.79 (m, 3H).
Example 61
5-(4-Chlorophenyl)-2-({1-(2,3-dichlorophenyl)-5-[(1RS)-1-hydroxyethyl]-1H--
1,2,4-triazol-3-yl}-methyl)-4-[(2S)-3,3,3-trifluoro-2-hydroxypropyl]-2,4-d-
ihydro-3H-1,2,4-triazol-3-one (diastereomeric mixture)
##STR00045##
[0439] To a solution of
5-(4-chlorophenyl)-2-({5-[(1RS)-1-hydroxyethyl]-1H-1,2,4-triazol-3-yl}met-
hyl)-4-[(2S)-3,3,3-trifluoro-2-hydroxypropyl]-2,4-dihydro-3H-1,2,4-triazol-
-3-one (500 mg, 0.92 mmol, 80% purity) in pyridine (12 ml) were
added (2,3-dichlorophenyl)boronic acid (176.36 mg, 0.92 mmol) and
copper(II) acetate (335.75 mg, 1.85 mmol). The reaction mixture was
heated to 60.degree. C. for 1 h and then stirred at room
temperature for 24 h, after which extra boronic acid (80 mg, 0.42
mmol) was added due to incomplete conversion. The reaction mixture
was further stirred at room temperature for 5 days. Over this time,
two additional portions of boronic acid (160 mg in total, 0.84
mmol) were added. After this, the resulting reaction mixture was
concentrated in vacuo, then diluted with MTBE and quenched with
aqueous hydrochloric acid (0.5 M). After phase separation, the
aqueous phase was extracted twice with MTBE. The combined organic
phases were dried over sodium sulfate, filtered, and concentrated
in vacuo. The crude product was purified by preparative HPLC
[method 4], and the desired compound (148 mg, 0.25 mmol, 97.3%
purity) was obtained as a mixture of diastereomers (yield 27%).
[0440] LC/MS [method 2]: R.sub.t=3.19 min; MS [ESIpos]: m/z=577
(M+H).sup.+
[0441] .sup.1H NMR (400 MHz, DMSO-d.sub.6): .delta. [ppm] 1.39 (d,
3H), 3.85 (dd, 1H), 4.00 (dd, 1H), 4.24-4.35 (m, 1H), 4.60-4.71 (m,
1H), 5.02-5.13 (m, 2H), 5.52 (br. s, 1H), 6.89 (dd, 1H), 7.55 (t,
1H), 7.59-7.66 (m, 3H), 7.73-7.78 (m, 2H), 7.87 (dd, 1H).
[0442] The two diastereomers were separated by preparative chiral
HPLC (SFC) [sample preparation: 141 mg dissolved in 18 ml methanol;
injection volume: 0.3 ml; column: Daicel Chiralcel.RTM. OX-H 5
.mu.m, 250.times.20 mm; eluent: carbon dioxide/methanol 70:30; flow
rate: 80 ml/min; temperature: 40.degree. C.; UV detection: 210 nm].
After separation, 58.5 mg of diastereomer 1 (Example 62), which
eluted first, and 53 mg of diastereomer 2 (Example 63), which
eluted later, were isolated.
Example 62
5-(4-Chlorophenyl)-2-{[1-(2,3-dichlorophenyl)-5-(1-hydroxyethyl)-1H-1,2,4--
triazol-3-yl]methyl}-4-[(2S)-3,3,3-trifluoro-2-hydroxypropyl]-2,4-dihydro--
3H-1,2,4-triazol-3-one (diastereomer 1)
[0443] LC/MS [method 2]: R.sub.t=3.21 min; MS [ESIpos]: m/z=577
(M+H).sup.+; 95% purity
[0444] Analytical chiral HPLC (SFC): R.sub.t=3.09 min, d.e.=100%
[column: Daicel Chiralcel.RTM. OX-3 250.times.4 mm; eluent: carbon
dioxide/methanol (5%.fwdarw.60%); flow rate: 3 ml/min; UV
detection: 220 nm].
[0445] .sup.1H NMR (400 MHz, DMSO-d.sub.6): .delta. [ppm] 1.39 (d,
3H), 3.85 (dd, 1H), 4.00 (dd, 1H), 4.24-4.36 (m, 1H), 4.65 (br. s,
1H), 5.01-5.13 (m, 2H), 5.52 (d, 1H), 6.89 (d, 1H), 7.55 (t, 1H),
7.59-7.66 (m, 3H), 7.72-7.78 (m, 2H), 7.87 (dd, 1H).
Example 63
5-(4-Chlorophenyl)-2-{[1-(2,3-dichlorophenyl)-5-(1-hydroxyethyl)-1H-1,2,4--
triazol-3-yl]methyl}-4-[(2S)-3,3,3-trifluoro-2-hydroxypropyl]-2,4-dihydro--
3H-1,2,4-triazol-3-one (diastereomer 2)
[0446] LC/MS [method 2]: R.sub.t=3.20 min; MS [ESIpos]: m/z=577
(M+H).sup.+; 95% purity
[0447] Analytical chiral HPLC (SFC): R.sub.t=3.38 min, d.e.=100%
[column: Daicel Chiralcel.RTM. OX-3 250.times.4 mm; eluent: carbon
dioxide/methanol (5%.fwdarw.60%); flow rate: 3 ml/min; UV
detection: 220 nm].
[0448] .sup.1H NMR (400 MHz, DMSO-d.sub.6): .delta. [ppm] 1.39 (d,
3H), 3.85 (dd, 1H), 4.00 (dd, 1H), 4.24-4.35 (m, 1H), 4.65 (br. s,
1H), 5.07 (s, 2H), 5.52 (d, 1H), 6.90 (d, 1H), 7.55 (t, 1H),
7.59-7.66 (m, 3H), 7.72-7.79 (m, 2H), 7.87 (dd, 1H).
Example 64
5-(4-Chlorophenyl)-2-({1-(2,3-difluorophenyl)-5-[(1RS)-1-hydroxyethyl]-1H--
1,2,4-triazol-3-yl}-methyl)-4-[(2S)-3,3,3-trifluoro-2-hydroxypropyl]-2,4-d-
ihydro-3H-1,2,4-triazol-3-one (diastereomeric mixture)
##STR00046##
[0450] To a solution of
5-(4-chlorophenyl)-2-({5-[(1RS)-1-hydroxyethyl]-1H-1,2,4-triazol-3-yl}met-
hyl)-4-[(2S)-3,3,3-trifluoro-2-hydroxypropyl]-2,4-dihydro-3H-1,2,4-triazol-
-3-one (430 mg, 0.99 mmol) in pyridine (12.5 ml) were added
(2,3-difluorophenyl)boronic acid (156.89 mg, 0.99 mmol) and
copper(II) acetate (360.94 mg, 1.99 mmol). The reaction mixture was
heated to 60.degree. C. for 1 h and then stirred at room
temperature for 24 h, after which extra boronic acid (80 mg, 0.51
mmol) was added due to incomplete conversion. The reaction mixture
was further stirred at room temperature for five days. Over this
time, five additional portions of boronic acid (400 mg in total,
2.54 mmol) were added. After this, the resulting reaction mixture
was concentrated in vacuo, then diluted with MTBE and quenched with
aqueous hydrochloric acid (0.5 M). After phase separation, the
aqueous phase was extracted twice with MTBE. The combined organic
phases were dried over sodium sulfate, filtered, and concentrated
in vacuo. The crude product was purified by preparative HPLC
[method 4], and the desired compound (44 mg, 0.08 mmol) was
obtained as a mixture of diastereomers (yield 8.1%).
[0451] LC/MS [method 2]: R.sub.t=2.97 min; MS [ESIpos]: m/z=545
(M+H).sup.+
[0452] .sup.1H NMR (400 MHz, DMSO-d.sub.6): .delta. [ppm] 1.42 (d,
3H), 3.85 (dd, 1H), 4.00 (dd, 1H), 4.30 (br. s, 1H), 4.76 (q, 1H),
5.02-5.13 (m, 2H), 6.89 (br. s, 1H), 7.35-7.43 (m, 1H), 7.45-7.51
(m, 1H), 7.59-7.71 (m, 3H), 7.72-7.79 (m, 2H).
[0453] The two diastereomers were separated by preparative chiral
HPLC [sample preparation: 40 mg dissolved in 1 ml ethanol;
injection volume: 0.5 ml; column: Daicel Chiralcel.RTM. OX-H 5
.mu.m, 250.times.20 mm; eluent: isohexane/ethanol 80:20; flow rate:
15 ml/min; temperature: 35.degree. C.; UV detection: 220 nm]. After
separation, 18 mg of diastereomer 1 (Example 65), which eluted
first, and 16 mg of diastereomer 2 (Example 66), which eluted
later, were isolated.
Example 65
5-(4-Chlorophenyl)-2-{[1-(2,3-difluorophenyl)-5-(1-hydroxyethyl)-1H-1,2,4--
triazol-3-yl]methyl}-4-[(2S)-3,3,3-trifluoro-2-hydroxypropyl]-2,4-dihydro--
3H-1,2,4-triazol-3-one (diastereomer 1)
[0454] Analytical chiral HPLC: R.sub.t=5.74 min, d.e.=100% [column:
Daicel Chiralcel.RTM. OX-H 5 .mu.m, 250.times.4.6 mm; eluent:
isohexane/ethanol 70:30+0.2% TFA and 1% water; flow rate: 1 ml/min;
temperature: 35.degree. C.; UV detection: 220 nm].
[0455] .sup.1H NMR (400 MHz, DMSO-d.sub.6): .delta. [ppm] 1.42 (d,
3H), 3.85 (dd, 1H), 4.00 (dd, 1H), 4.24-4.35 (m, 1H), 4.76 (quin,
1H), 5.02-5.13 (m, 2H), 5.58 (d, 1H), 6.89 (d, 1H), 7.35-7.44 (m,
1H), 7.45-7.52 (m, 1H), 7.59-7.72 (m, 3H), 7.72-7.78 (m, 2H).
Example 66
5-(4-Chlorophenyl)-2-{[1-(2,3-difluorophenyl)-5-(1-hydroxyethyl)-1H-1,2,4--
triazol-3-yl]methyl}-4-[(2S)-3,3,3-trifluoro-2-hydroxypropyl]-2,4-dihydro--
3H-1,2,4-triazol-3-one (diastereomer 2)
[0456] Analytical chiral HPLC: R.sub.t=6.59 min, d.e.=99.2%
[column: Daicel Chiralcel.RTM. OX-H 5 .mu.m, 250.times.4.6 mm;
eluent: isohexane/ethanol 70:30+0.2% TFA and 1% water; flow rate: 1
ml/min; temperature: 35.degree. C.; UV detection: 220 nm].
[0457] .sup.1H NMR (400 MHz, DMSO-d.sub.6): .delta. [ppm] 1.42 (d,
3H), 3.85 (dd, 1H), 4.00 (dd, 1H), 4.23-4.36 (m, 1H), 4.76 (quin,
1H), 5.07 (s, 2H), 5.58 (d, 1H), 6.90 (d, 1H), 7.35-7.43 (m, 1H),
7.44-7.51 (m, 1H), 7.59-7.72 (m, 3H), 7.72-7.78 (m, 2H).
Example 67
2-{[1-(2-Chloro-3-fluorophenyl)-5-(1-hydroxyethyl)-1H-H-1,2,4-triazol-3-yl-
]methyl}-5-(4-chlorophenyl)-4-[(2S)-3,3,3-trifluoro-2-hydroxypropyl]-2,4-d-
ihydro-3H-1,2,4-triazol-3-one (diastereomer 1)
##STR00047##
[0459] To a solution of
5-(4-chlorophenyl)-2-({5-[(1RS)-1-hydroxyethyl]-1H-1,2,4-triazol-3-yl}met-
hyl)-4-[(2S)-3,3,3-trifluoro-2-hydroxypropyl]-2,4-dihydro-3H-1,2,4-triazol-
-3-one (500 mg, 0.92 mmol, 80% purity) in pyridine (12 ml) were
added (2-chloro-3-fluorophenyl)boronic acid (161.15 mg, 0.92 mmol)
and copper(II) acetate (335.75 mg, 1.85 mmol). The reaction mixture
was heated to 60.degree. C. for 1 h and then stirred at room
temperature for 24 h, after which extra boronic acid (75 mg, 0.43
mmol) was added due to incomplete conversion. The reaction mixture
was further stirred at room temperature for six days. Over this
time, five additional portions of boronic acid (375 mg in total,
2.15 mmol) were added. After this, the resulting reaction mixture
was concentrated in vacuo, then diluted with MTBE and quenched with
aqueous hydrochloric acid (0.5 M). After phase separation, the
aqueous phase was extracted twice with MTBE. The combined organic
phases were dried over sodium sulfate, filtered, and concentrated
in vacuo. The crude product was purified by preparative HPLC
[method 4], and 91 mg of the desired compound as a diastereomeric
mixture still containing some impurities were isolated.
[0460] A further purification by preparative chiral HPLC yielded
the two pure, separated diastereomers [sample preparation: 90 mg
dissolved in 3 ml ethanol; injection volume: 0.3 ml; column: Daicel
Chiralcel.RTM. OX-H 5 .mu.m, 250.times.20 mm; eluent:
isohexane/ethanol 80:20; flow rate: 15 ml/min; temperature:
35.degree. C.; UV detection: 220 nm]. After separation, 20 mg of
diastereomer 1 (Example 67), which eluted first, and 21 mg of
diastereomer 2 (Example 68), which eluted later, were isolated.
[0461] LC/MS [method 2]: R.sub.t=3.01 min; MS [ESIpos]: m/z=561
(M+H).sup.+
[0462] Analytical chiral HPLC: R.sub.t=6.22 min, d.e.=100% [column:
Daicel Chiralcel.RTM. OX-H 5 .mu.m, 250.times.4.6 mm; eluent:
isohexane/ethanol 70:30+0.2% TFA and 1% water; flow rate: 1 ml/min;
temperature: 35.degree. C.; UV detection: 220 nm].
[0463] .sup.1H NMR (400 MHz, DMSO-d.sub.6): .delta. [ppm] 1.39 (d,
3H), 3.85 (dd, 1H), 4.00 (dd, 1H), 4.23-4.36 (m, 1H), 4.66 (quin,
1H), 5.01-5.14 (m, 2H), 5.53 (d, 1H), 6.89 (d, 1H), 7.48-7.70 (m,
5H), 7.72-7.78 (m, 2H).
Example 68
2-{[1-(2-Chloro-3-fluorophenyl)-5-(1-hydroxyethyl)-1H-H-1,2,4-triazol-3-yl-
]methyl}-5-(4-chlorophenyl)-4-[(2S)-3,3,3-trifluoro-2-hydroxypropyl]-2,4-d-
ihydro-3H-1,2,4-triazol-3-one (diastereomer 2)
[0464] LC/MS [method 2]: R.sub.t=3.01 min; MS [ESIpos]: m/z=561
(M+H).sup.+
[0465] Analytical chiral HPLC: R.sub.t=7.94 min, d.e.=100% [column:
Daicel Chiralcel.RTM. OX-H 5 .mu.m, 250.times.4.6 mm; eluent:
isohexane/ethanol 70:30+0.2% TFA and 1% water; flow rate: 1 ml/min;
temperature: 35.degree. C.; UV detection: 220 nm].
[0466] .sup.1H NMR (400 MHz, DMSO-d.sub.6): .delta. [ppm] 1.40 (d,
3H), 3.85 (dd, 1H), 4.00 (dd, 1H), 4.24-4.36 (m, 1H), 4.66 (quin,
1H), 5.07 (s, 2H), 5.53 (d, 1H), 6.90 (d, 1H), 7.49-7.70 (m, 5H),
7.72-7.78 (m, 2H).
Example 69
5-(4-Chlorophenyl)-2-({5-[(1RS)-1-hydroxyethyl]-1-(2-methylphenyl)-1H-1,2,-
4-triazol-3-yl}-methyl)-4-[(2S)-3,3,3-trifluoro-2-hydroxypropyl]-2,4-dihyd-
ro-3H-1,2,4-triazol-3-one (diastereomeric mixture)
##STR00048##
[0468] To a solution of
5-(4-chlorophenyl)-2-({5-[(1RS)-1-hydroxyethyl]-1H-1,2,4-triazol-3-yl}met-
hyl)-4-[(2S)-3,3,3-trifluoro-2-hydroxypropyl]-2,4-dihydro-3H-1,2,4-triazol-
-3-one (400 mg, 0.92 mmol) in pyridine (12 ml) were added
(2-methylphenyl)boronic acid (251.32 mg, 1.85 mmol) and copper(II)
acetate (335.75 mg, 1.85 mmol). The reaction mixture was stirred at
room temperature for 5 days, after which extra boronic acid (62.8
mg, 0.46 mmol, 0.5 eq.) was added due to incomplete conversion.
After stirring for two additional days, the reaction mixture was
concentrated in vacuo, then diluted with MTBE and quenched with
aqueous hydrochloric acid (0.5 M). After phase separation, the
aqueous phase was extracted twice with MTBE. The combined organic
phases were dried over sodium sulfate, filtered, and concentrated
in vacuo. The crude product was purified by preparative HPLC
[method 4], and the desired compound (100 mg, 0.17 mmol) was
obtained as a mixture of diastereomers (yield 17.2%, 90%
purity).
[0469] LC/MS [method 3]: R.sub.t=1.24 min; MS [ESIpos]: m/z=523
(M+H).sup.+
[0470] The two diastereomers were separated by preparative chiral
HPLC [sample preparation: 98 mg dissolved in 2 ml ethanol/isohexane
(1:1); injection volume: 1 ml; column: Daicel Chiralcel.RTM. OX-H 5
.mu.m, 250.times.20 mm; eluent: isohexane/ethanol 75:25; flow rate:
15 ml/min; temperature: 30.degree. C.; UV detection: 220 nm]. After
separation, 37 mg of diastereomer 1 (Example 70), which eluted
first, and 39 mg of diastereomer 2 (Example 71), which eluted
later, were isolated.
Example 70
5-(4-Chlorophenyl)-2-{[5-(1-hydroxyethyl)-1-(2-methylphenyl)-1H-1,2,4-tria-
zol-3-yl]methyl}-4-[(2S)-3,3,3-trifluoro-2-hydroxypropyl]-2,4-dihydro-3H-1-
,2,4-triazol-3-one (diastereomer 1)
[0471] LC/MS [method 3]: R.sub.t=1.24 min; MS [ESIpos]: m/z=523
(M+H).sup.+
[0472] Analytical chiral HPLC: R.sub.t=7.65 min, d.e.=100% [column:
LUX Cellulose-4, 5 .mu.m, 250.times.4.6 mm; eluent:
isohexane/ethanol 70:30; flow rate: 1 ml/min; temperature:
40.degree. C.; UV detection: 220 nm].
[0473] .sup.1H NMR (400 MHz, DMSO-d.sub.6): .delta. [ppm] 1.36 (d,
3H), 1.98 (s, 3H), 3.85 (dd, 1H), 4.00 (dd, 1H), 4.23-4.35 (m, 1H),
4.54 (quin, 1H), 5.00-5.12 (m, 2H), 5.48 (d, 1H), 6.89 (d, 1H),
7.31-7.49 (m, 4H), 7.59-7.65 (m, 2H), 7.71-7.77 (m, 2H).
Example 71
5-(4-Chlorophenyl)-2-{[5-(1-hydroxyethyl)-1-(2-methylphenyl)-1H-1,2,4-tria-
zol-3-yl]methyl}-4-[(2S)-3,3,3-trifluoro-2-hydroxypropyl]-2,4-dihydro-3H-1-
,2,4-triazol-3-one (diastereomer 2)
[0474] Analytical chiral HPLC: R.sub.t=10.27 min, d.e.=100%
[column: LUX Cellulose-4, 5 .mu.m, 250.times.4.6 mm; eluent:
isohexane/ethanol 70:30; flow rate: 1 ml/min; temperature:
40.degree. C.; UV detection: 220 nm].
[0475] .sup.1H NMR (400 MHz, DMSO-d.sub.6): .delta. [ppm] 1.37 (d,
3H), 1.98 (s, 3H), 3.84 (dd, 1H), 4.00 (dd, 1H), 4.24-4.35 (m, 1H),
4.54 (quin, 1H), 5.06 (s, 2H), 5.48 (d, 1H), 6.90 (d, 1H),
7.31-7.49 (m, 4H), 7.58-7.66 (m, 2H), 7.71-7.78 (m, 2H).
Example 72
5-(4-Chlorophenyl)-2-({5-[(RS)-1-hydroxyethyl]-1-[2-(trifluoromethyl)pheny-
l]-H-1,2,4-triazol-3-yl}methyl)-4-[(2S)-3,3,3-trifluoro-2-hydroxypropyl]-2-
,4-dihydro-3H-1,2,4-triazol-3-one (diastereomeric mixture)
##STR00049##
[0477] To a solution of
5-(4-chlorophenyl)-2-({5-[(1RS)-1-hydroxyethyl]-1H-1,2,4-triazol-3-yl}met-
hyl)-4-[(2S)-3,3,3-trifluoro-2-hydroxypropyl]-2,4-dihydro-3H-1,2,4-triazol-
-3-one (600 mg, 1.11 mmol, 80% purity) in pyridine (14.5 ml) were
added [2-(trifluoromethyl)phenyl]boronic acid (421.30 mg, 2.22
mmol) and copper(II) acetate (402.9 mg, 2.22 mmol). The reaction
mixture was heated to 60.degree. C. for 2 h and then stirred at
room temperature for 5 days, after which extra boronic acid (105
mg, 0.55 mmol, 0.5 eq.) was added due to incomplete conversion.
After further stirring at room temperature overnight, the resulting
reaction mixture was concentrated in vacuo, then diluted with MTBE
and quenched with aqueous hydrochloric acid (0.5 M). After phase
separation, the aqueous phase was extracted twice with MTBE. The
combined organic phases were dried over sodium sulfate, filtered,
and concentrated in vacuo. The crude product was purified by
preparative HPLC [method 4], and the desired compound (80 mg) was
obtained as a mixture of diastereomers (yield 12.4%).
[0478] LC/MS [method 2]: R.sub.t=3.19 min; MS [ESIpos]: m/z=577
(M+H).sup.+
[0479] The two diastereomers were separated by preparative chiral
HPLC [sample preparation: 78 mg dissolved in 2 ml ethanol/isohexane
(1:1); injection volume: 1 ml; column: Daicel Chiralcel.RTM. OX-H 5
.mu.m, 250.times.20 mm; eluent: isohexane/ethanol 75:25; flow rate:
15 ml/min; temperature: 30.degree. C.; UV detection: 220 nm]. After
separation, 34 mg of diastereomer 1 (Example 73), which eluted
first, and 30 mg of diastereomer 2 (Example 74), which eluted
later, were isolated.
Example 73
5-(4-Chlorophenyl)-2-({5-(1-hydroxyethyl)-1-[2-(trifluoromethyl)phenyl]-H--
1,2,4-triazol-3-yl}-methyl)-4-[(2S)-3,3,3-trifluoro-2-hydroxypropyl]-2,4-d-
ihydro-3H-1,2,4-triazol-3-one (diastereomer 1)
[0480] Analytical chiral HPLC: R.sub.t=6.16 min, d.e.=100% [column:
LUX Cellulose-4, 5 .mu.m, 250.times.4.6 mm; eluent:
isohexane/ethanol 70:30; flow rate: 1 ml/min; temperature:
40.degree. C.; UV detection: 220 nm].
[0481] .sup.1H NMR (400 MHz, DMSO-d.sub.6): .delta. [ppm] 1.36 (d,
3H), 3.84 (dd, 1H), 4.00 (dd, 1H), 4.24-4.35 (m, 1H), 4.57 (quin,
1H), 4.99-5.12 (m, 2H), 5.50 (d, 1H), 6.89 (d, 1H), 7.59-7.65 (m,
2H), 7.66-7.71 (m, 1H), 7.72-7.76 (m, 2H), 7.77-7.90 (m, 2H),
7.93-7.99 (m, 1H).
Example 74
5-(4-Chlorophenyl)-2-({5-(1-hydroxyethyl)-1-[2-(trifluoromethyl)phenyl]-H--
1,2,4-triazol-3-yl}-methyl)-4-[(2S)-3,3,3-trifluoro-2-hydroxypropyl]-2,4-d-
ihydro-3H-1,2,4-triazol-3-one (diastereomer 2)
[0482] Analytical chiral HPLC: R.sub.t=8.67 min, d.e.=100% [column:
LUX Cellulose-4, 5 .mu.m, 250.times.4.6 mm; eluent:
isohexane/ethanol 70:30; flow rate: 1 ml/min; temperature:
40.degree. C.; UV detection: 220 nm].
[0483] .sup.1H NMR (400 MHz, DMSO-d.sub.6): .delta. [ppm] 1.36 (d,
3H), 3.84 (dd, 1H), 3.99 (dd, 1H), 4.24-4.35 (m, 1H), 4.54-4.62 (m,
1H), 5.05 (s, 2H), 5.50 (d, 1H), 6.90 (d, 1H), 7.60-7.65 (m, 2H),
7.67-7.71 (m, 1H), 7.72-7.90 (m, 4H), 7.93-7.98 (m, 1H).
Example 75
5-(4-Chlorophenyl)-2-({1-(3-fluorophenyl)-5-[(1RS)-1-hydroxyethyl]-1H-1,2,-
4-triazol-3-yl}-methyl)-4-[(2S)-3,3,3-trifluoro-2-hydroxypropyl]-2,4-dihyd-
ro-3H-1,2,4-triazol-3-one (diastereomeric mixture)
##STR00050##
[0485] To a solution of
5-(4-chlorophenyl)-2-({5-[(1RS)-1-hydroxyethyl]-1H-1,2,4-triazol-3-yl}met-
hyl)-4-[(2S)-3,3,3-trifluoro-2-hydroxypropyl]-2,4-dihydro-3H-1,2,4-triazol-
-3-one (430 mg, 0.795 mmol, 80% purity) in pyridine (10 ml) were
added (3-fluorophenyl)boronic acid (222.432 mg, 1.59 mmol) and
copper(II) acetate (288.75 mg, 1.59 mmol). The reaction mixture was
heated to 60.degree. C. for 2 h and then stirred at room
temperature for 5 days, after which extra boronic acid (55.6 mg,
0.40 mmol) was added due to incomplete conversion. The reaction
mixture was again heated to 60.degree. C. for 2 h, followed by
stirring at room temperature overnight. The resulting reaction
mixture was concentrated in vacuo, then diluted with MTBE and
quenched with aqueous hydrochloric acid (0.5 M). After phase
separation, the aqueous phase was extracted twice with MTBE. The
combined organic phases were dried over sodium sulfate, filtered,
and concentrated in vacuo. The crude product was purified by
preparative HPLC [method 4], and the desired compound (100 mg, 0.19
mmol) was obtained as a mixture of diastereomers (yield 23.9%).
[0486] LC/MS [method 2]: R.sub.t=2.99 min; MS [ESIpos]: m/z=527
(M+H).sup.+
[0487] .sup.1H NMR (400 MHz, DMSO-d.sub.6): .delta. [ppm] 1.47 (d,
3H), 3.85 (dd, 1H), 4.01 (dd, 1H), 4.30 (br. s, 1H), 4.83 (q, 1H),
5.02-5.13 (m, 2H), 6.89 (br. s, 1H), 7.38 (td, 1H), 7.48-7.66 (m,
5H), 7.72-7.78 (m, 2H).
[0488] The two diastereomers were separated by preparative chiral
HPLC [sample preparation: 97 mg dissolved in 4 ml ethanol/isohexane
(1:1); injection volume: 1 ml; column: Daicel Chiralcel.RTM. OX-H 5
.mu.m, 250.times.20 mm; eluent: isohexane/ethanol 80:20; flow rate:
15 ml/min; temperature: 30.degree. C.; UV detection: 220 nm]. After
separation, 36 mg of (1S)-diastereomer (Example 76), which eluted
first, and 40 mg of (1R)-diastereomer (Example 77), which eluted
later, were isolated.
Example 76
5-(4-Chlorophenyl)-2-({1-(3-fluorophenyl)-5-[(1
S)-1-hydroxyethyl]-1H-1,2,4-triazol-3-yl}methyl)-4-[(2S)-3,3,3-trifluoro--
2-hydroxypropyl]-2,4-dihydro-3H-1,2,4-triazol-3-one
##STR00051##
[0490] LC/MS [method 3]: R.sub.t=1.24 min; MS [ESIpos]: m/z=527
(M+H).sup.+
[0491] Analytical chiral HPLC: R.sub.t=9.71 min, d.e.=100% [column:
LUX Cellulose-4, 5 .mu.m, 250.times.4.6 mm; eluent:
isohexane/ethanol 80:20; flow rate: 1 ml/min; temperature:
40.degree. C.; UV detection: 220 nm].
[0492] .sup.1H NMR (400 MHz, DMSO-d.sub.6): .delta. [ppm] 1.47 (d,
3H), 3.85 (dd, 1H), 4.00 (dd, 1H), 4.23-4.37 (m, 1H), 4.82 (quin,
1H), 5.01-5.13 (m, 2H), 5.76 (d, 1H), 6.89 (d, 1H), 7.38 (td, 1H),
7.48-7.66 (m, 5H), 7.72-7.79 (m, 2H).
[0493] The absolute stereochemistry of the compound was determined
by additionally performing the same reaction with enantiopure
diastereomer
5-(4-chlorophenyl)-2-({5-[(1S)-1-hydroxyethyl]-1H-1,2,4-triazol-3-yl}meth-
yl)-4-[(2S)-3,3,3-trifluoro-2-hydroxypropyl]-2,4-dihydro-3H-1,2,4-triazol--
3-one (Example 5A) as the starting material and comparison of the
two respective products by analytical chiral HPLC.
Example 77
5-(4-Chlorophenyl)-2-({1-(3-fluorophenyl)-5-[(1R)-1-hydroxyethyl]-1H-1,2,4-
-triazol-3-yl}methyl)-4-[(2S)-3,3,3-trifluoro-2-hydroxypropyl]-2,4-dihydro-
-3H-1,2,4-triazol-3-one
##STR00052##
[0495] LC/MS [method 2]: R.sub.t=2.93 min; MS [ESIpos]: m/z=527
(M+H).sup.+
[0496] Analytical chiral HPLC: R.sub.t=13.60 min, d.e.=100%
[column: LUX Cellulose-4, 5 .mu.m, 250.times.4.6 mm; eluent:
isohexane/ethanol 80:20; flow rate: 1 ml/min; temperature:
40.degree. C.; UV detection: 220 nm].
[0497] .sup.1H NMR (400 MHz, DMSO-d.sub.6): .delta. [ppm] 1.47 (d,
3H), 3.85 (dd, 1H), 4.01 (dd, 1H), 4.24-4.36 (m, 1H), 4.83 (quin,
1H), 5.07 (s, 2H), 5.76 (d, 1H), 6.90 (d, 1H), 7.38 (td, 1H),
7.48-7.65 (m, 5H), 7.72-7.78 (m, 2H).
[0498] The absolute stereochemistry of the compound was determined
by additionally performing the same reaction with enantiopure
diastereomer
5-(4-chlorophenyl)-2-({5-[(1R)-1-hydroxyethyl]-1H-1,2,4-triazol-3-yl}meth-
yl)-4-[(2S)-3,3,3-trifluoro-2-hydroxypropyl]-2,4-dihydro-3H-1,2,4-triazol--
3-one (Example 6A) as the starting material and comparison of the
two respective products by analytical chiral HPLC.
Example 78
5-(4-Chlorophenyl)-2-({1-(3-chlorophenyl)-5-[(1RS)-1-hydroxyethyl]-1H-1,2,-
4-triazol-3-yl}-methyl)-4-[(2S)-3,3,3-trifluoro-2-hydroxypropyl]-2,4-dihyd-
ro-3H-1,2,4-triazol-3-one (diastereomeric mixture)
##STR00053##
[0500] To a solution of
5-(4-chlorophenyl)-2-({5-[(1RS)-1-hydroxyethyl]-1H-1,2,4-triazol-3-yl}met-
hyl)-4-[(2S)-3,3,3-trifluoro-2-hydroxypropyl]-2,4-dihydro-3H-1,2,4-triazol-
-3-one (430 mg, 0.795 mmol, 80% purity) in pyridine (10 ml) were
added (3-chlorophenyl)boronic acid (248.59 mg, 1.59 mmol) and
copper(II) acetate (288.75 mg, 1.59 mmol). The reaction mixture was
heated to 60.degree. C. for 2 h and then stirred at room
temperature for 5 days, after which extra boronic acid (62.1 mg,
0.40 mmol) was added due to incomplete conversion. The reaction
mixture was again heated to 60.degree. C. for 2 h, followed by
stirring at room temperature overnight. The resulting reaction
mixture was concentrated in vacuo, then diluted with MTBE and
quenched with aqueous hydrochloric acid (0.5 M). After phase
separation, the aqueous phase was extracted twice with MTBE. The
combined organic phases were dried over sodium sulfate, filtered,
and concentrated in vacuo. The crude product was purified by
preparative HPLC [method 4], and the desired compound (130 mg, 0.24
mmol) was obtained as a mixture of diastereomers (yield 30.1%).
[0501] LC/MS [method 2]: R.sub.t=3.19 min; MS [ESIpos]: m/z=543
(M+H).sup.+
[0502] .sup.1H NMR (400 MHz, DMSO-d.sub.6): .delta. [ppm] 1.47 (d,
3H), 3.85 (dd, 1H), 4.01 (dd, 1H), 4.30 (br. s, 1H), 4.81 (q, 1H),
5.02-5.13 (m, 2H), 6.89 (br. s, 1H), 7.56-7.67 (m, 5H), 7.72-7.79
(m, 3H).
[0503] The two diastereomers were separated by preparative chiral
HPLC [sample preparation: 128 mg dissolved in 4 ml
ethanol/isohexane (1:1); injection volume: 1 ml; column: Daicel
Chiralcel.RTM. OX-H 5 .mu.m, 250.times.20 mm; eluent:
isohexane/ethanol 80:20; flow rate: 15 ml/min; temperature:
30.degree. C.; UV detection: 220 nm]. After separation, 52 mg of
(1S)-diastereomer (Example 79), which eluted first, and 49 mg of
(1R)-diastereomer (Example 80), which eluted later, were
isolated.
Example 79
5-(4-Chlorophenyl)-2-({1-(3-chlorophenyl)-5-[(1S)-1-hydroxyethyl]-1H-1,2,4-
-triazol-3-yl}methyl)-4-[(2S)-3,3,3-trifluoro-2-hydroxypropyl]-2,4-dihydro-
-3H-1,2,4-triazol-3-one
##STR00054##
[0505] LC/MS [method 2]: R.sub.t=3.14 min; MS [ESIpos]: m/z=543
(M+H).sup.+
[0506] Analytical chiral HPLC: R.sub.t=9.96 min, d.e.=100% [column:
LUX Cellulose-4, 5 .mu.m, 250.times.4.6 mm; eluent:
isohexane/ethanol 80:20; flow rate: 1 ml/min; temperature:
35.degree. C.; UV detection: 220 nm].
[0507] .sup.1H NMR (400 MHz, DMSO-d.sub.6): .delta. [ppm] 1.47 (d,
3H), 3.85 (dd, 1H), 4.01 (dd, 1H), 4.23-4.36 (m, 1H), 4.81 (quin,
1H), 5.01-5.13 (m, 2H), 5.76 (d, 1H), 6.89 (d, 1H), 7.56-7.66 (m,
5H), 7.71-7.79 (m, 3H).
[0508] .sup.13C NMR (125 MHz, DMSO-d.sub.6): .delta. [ppm] 21.3,
42.1, 42.2, 59.6, 65.5, 123.0, 124.5, 124.6, 125.3, 128.5, 128.9
(2.times.), 130.0 (2.times.), 130.7, 133.0, 135.2, 138.2, 144.8,
153.1, 157.8, 158.6.
[0509] The absolute stereochemistry of the compound was determined
by additionally performing the same reaction with enantiopure
diastereomer
5-(4-chlorophenyl)-2-({5-[(1S)-1-hydroxyethyl]-1H-1,2,4-triazol-3-yl}meth-
yl)-4-[(2S)-3,3,3-trifluoro-2-hydroxypropyl]-2,4-dihydro-3H-1,2,4-triazol--
3-one (Example 5A) as the starting material and comparison of the
two respective products by analytical chiral HPLC.
Example 80
5-(4-Chlorophenyl)-2-({1-(3-chlorophenyl)-5-[(1R)-1-hydroxyethyl]-1H-1,2,4-
-triazol-3-yl}-methyl)-4-[(2S)-3,3,3-trifluoro-2-hydroxypropyl]-2,4-dihydr-
o-3H-1,2,4-triazol-3-one
##STR00055##
[0511] LC/MS [method 2]: R.sub.t=3.15 min; MS [ESIpos]: m/z=543
(M+H).sup.+
[0512] Analytical chiral HPLC: R.sub.t=14.41 min, d.e.=100%
[column: LUX Cellulose-4, 5 .mu.m, 250.times.4.6 mm; eluent:
isohexane/ethanol 80:20; flow rate: 1 ml/min; temperature:
35.degree. C.; UV detection: 220 nm].
[0513] .sup.1H NMR (400 MHz, DMSO-d.sub.6): .delta. [ppm] 1.47 (d,
3H), 3.85 (dd, 1H), 4.01 (dd, 1H), 4.24-4.37 (m, 1H), 4.81 (quin,
1H), 5.07 (s, 2H), 5.76 (d, 1H), 6.90 (d, 1H), 7.56-7.66 (m, 5H),
7.71-7.79 (m, 3H).
[0514] The absolute stereochemistry of the compound was determined
by additionally performing the same reaction with enantiopure
diastereomer
5-(4-chlorophenyl)-2-({5-[(1R)-1-hydroxyethyl]-1H-1,2,4-triazol-3-yl
J}methyl)-4-[(2S)-3,3,3-trifluoro-2-hydroxypropyl]-2,4-dihydro-3H-1,2,4-t-
riazol-3-one (Example 6A) as the starting material and comparison
of the two respective products by analytical chiral HPLC.
Example 81
5-(4-Chlorophenyl)-2-({1-(2-chlorophenyl)-5-[(1RS)-1-hydroxyethyl]-1H-1,2,-
4-triazol-3-yl}-methyl)-4-[(2S)-3,3,3-trifluoro-2-hydroxypropyl]-2,4-dihyd-
ro-3H-1,2,4-triazol-3-one (diastereomeric mixture)
##STR00056##
[0516] To a solution of
5-(4-chlorophenyl)-2-({5-[(1RS)-1-hydroxyethyl]-1H-1,2,4-triazol-3-yl}met-
hyl)-4-[(2S)-3,3,3-trifluoro-2-hydroxypropyl]-2,4-dihydro-3H-1,2,4-triazol-
-3-one (2.10 g, 3.88 mmol, 80% purity) in pyridine (50 ml) were
added (2-chlorophenyl)boronic acid (1.214 g, 7.76 mmol) and
copper(II) acetate (1.410 g, 7.76 mmol). The reaction mixture was
heated to 60.degree. C. for 1 h and then stirred at room
temperature for 5 days, after which extra boronic acid (303 mg,
1.94 mmol) was added due to incomplete conversion. After stirring
at room temperature for two additional days, the resulting reaction
mixture was concentrated in vacuo, then diluted with MTBE and
quenched with aqueous hydrochloric acid (0.5 M). After phase
separation, the aqueous phase was extracted twice with MTBE. The
combined organic phases were dried over sodium sulfate, filtered,
and concentrated in vacuo. The crude product was purified by
preparative HPLC [method 4], and the desired compound (580 mg, 1.01
mmol, 95% purity) was obtained as a mixture of diastereomers (yield
26.1%).
[0517] LC/MS [method 3]: R.sub.t=1.24 min; MS [ESIpos]: m/z=543
(M+H).sup.+
[0518] .sup.1H NMR (400 MHz, DMSO-d.sub.6): .delta. [ppm] 1.38 (d,
3H), 3.85 (dd, 1H), 4.00 (dd, 1H), 4.30 (br. s, 1H), 4.55-4.64 (m,
1H), 5.01-5.13 (m, 2H), 6.85-6.94 (m, 1H), 7.50-7.65 (m, 5H),
7.67-7.78 (m, 3H).
[0519] The two diastereomers were separated by preparative chiral
HPLC (SFC) [sample preparation: 575 mg dissolved in 35 ml methanol;
injection volume: 0.4 ml; column: Daicel Chiralcel.RTM. OX-H 5
.mu.m, 250.times.20 mm; eluent: carbon dioxide/methanol 70:30; flow
rate: 80 ml/min; temperature: 40.degree. C.; UV detection: 210 nm].
After separation, 206 mg of (1S)-diastereomer (Example 82), which
eluted first, and 189 mg of (1R)-diastereomer (Example 83), which
eluted later, were isolated.
Example 82
5-(4-Chlorophenyl)-2-({1-(2-chlorophenyl)-5-[(1S)-1-hydroxyethyl]-1H-1,2,4-
-triazol-3-yl}methyl)-4-[(2S)-3,3,3-trifluoro-2-hydroxypropyl]-2,4-dihydro-
-3H-1,2,4-triazol-3-one
##STR00057##
[0521] LC/MS [method 3]: R.sub.t=1.24 min; MS [ESIpos]: m/z=543
(M+H).sup.+
[0522] Analytical chiral HPLC: R.sub.t=8.34 min, d.e.=100% [column:
LUX Cellulose-4, 5 .mu.m, 250.times.4.6 mm; eluent:
isohexane/ethanol 70:30; flow rate: 1 ml/min; temperature:
40.degree. C.; UV detection: 220 nm].
[0523] .sup.1H NMR (400 MHz, DMSO-d.sub.6): .delta. [ppm] 1.38 (d,
3H), 3.85 (dd, 1H), 4.00 (dd, 1H), 4.30 (br. s, 1H), 4.59 (q, 1H),
5.01-5.13 (m, 2H), 5.50 (br. s, 1H), 6.90 (d, 1H), 7.50-7.65 (m,
5H), 7.67-7.78 (m, 3H).
[0524] The absolute stereochemistry of the compound was determined
by additionally performing the same reaction with enantiopure
diastereomer
5-(4-chlorophenyl)-2-({5-[(1S)-1-hydroxyethyl]-1H-1,2,4-triazol-3-yl}meth-
yl)-4-[(2S)-3,3,3-trifluoro-2-hydroxypropyl]-2,4-dihydro-3H-1,2,4-triazol--
3-one (Example 5A) as the starting material and comparison of the
two respective products by analytical chiral HPLC.
Example 83
5-(4-Chlorophenyl)-2-({1-(2-chlorophenyl)-5-[(1R)-1-hydroxyethyl]-1H-1,2,4-
-triazol-3-yl}-methyl)-4-[(2S)-3,3,3-trifluoro-2-hydroxypropyl]-2,4-dihydr-
o-3H-1,2,4-triazol-3-one
##STR00058##
[0526] Analytical chiral HPLC: R.sub.t=11.88 min, d.e.=98.1%
[column: LUX Cellulose-4, 5 .mu.m, 250.times.4.6 mm; eluent:
isohexane/ethanol 70:30; flow rate: 1 ml/min; temperature:
40.degree. C.; UV detection: 220 nm].
[0527] .sup.1H NMR (400 MHz, DMSO-d.sub.6): .delta. [ppm] 1.38 (d,
3H), 3.85 (dd, 1H), 4.00 (dd, 1H), 4.24-4.36 (m, 1H), 4.54-4.65 (m,
1H), 5.07 (s, 2H), 5.51 (br. s, 1H), 6.90 (d, 1H), 7.50-7.65 (m,
5H), 7.68-7.79 (m, 3H).
[0528] The absolute stereochemistry of the compound was determined
by additionally performing the same reaction with enantiopure
diastereomer
5-(4-chlorophenyl)-2-({5-[(1R)-1-hydroxyethyl]-1H-1,2,4-triazol-3-yl
J}methyl)-4-[(2S)-3,3,3-trifluoro-2-hydroxypropyl]-2,4-dihydro-3H-1,2,4-t-
riazol-3-one (Example 6A) as the starting material and comparison
of the two respective products by analytical chiral HPLC.
B. EVALUATION OF BIOLOGICAL ACTIVITY
Abbreviations and Acronyms
[0529] Acc. No. accession number [0530] AVP arginine vasopressin
[0531] B.sub.max maximal ligand binding capacity [0532] BSA bovine
serum albumin [0533] cAMP cyclic adenosine monophosphate [0534]
Cat. No. catalogue number [0535] cDNA complementary
deoxyribonucleic acid [0536] CHO chinese hamster ovary [0537] CRE
cAMP response element [0538] Ct cycle threshold [0539] DMEM/F12
Dulbecco's modified Eagle's medium/Ham's F12 medium (1:1) [0540]
DNA deoxyribonucleic acid [0541] DTT dithiothreitol [0542]
EC.sub.50 half-maximal effective concentration [0543] EDTA
ethylenediamine-tetraacetic acid [0544] FAM carboxyfluorescein
succinimidyl ester [0545] f.c. final concentration [0546] FCS fetal
calf serum [0547] HEPES
4-(2-hydroxyethyl)piperazine-1-ethanesulfonic acid [0548] IC.sub.50
half-maximal inhibitory concentration [0549] K.sub.d dissociation
constant [0550] K.sub.i dissociation constant of an inhibitor
[0551] mRNA messenger ribonucleic acid [0552] PBS phosphate
buffered saline [0553] p.o. per os, peroral [0554] RNA ribonucleic
acid [0555] RTPCR real-time polymerase chain reaction [0556] SPA
scintillation proximity assay [0557] TAMRA
carboxytetramethylrhodamine [0558] TRIS
2-amino-2-hydroxymethylpropane-1,3-diol
[0559] Demonstration of the activity of the compounds of the
present invention may be accomplished through in vitro, ex vivo,
and in vivo assays that are well known in the art. For example, to
demonstrate the activity of the compounds of the present invention,
the following assays may be used.
B-1. Cellular In Vitro Assay for Determining Vasopressin Receptor
Activity
[0560] The identification of agonists and antagonists of the V1a
and V2 vasopressin receptors from humans, rats and dogs as well as
the quantification of the activity of the compounds of the
invention is carried out using recombinant cell lines. These cell
lines originally derive from a hamster's ovary epithelial cell
(Chinese Hamster Ovary, CHO K1, ATCC: American Type Culture
Collection, Manassas, Va. 20108, USA). The test cell lines
constitutively express the human, rat or dog V1a or V2 receptors.
In case of the G.sub..alpha.q-coupled V1a receptors, cells are also
stably transfected with a modified form of the calcium-sensitive
photoproteins aequorin (human and rat V1a) or obelin (dog V1a),
which, after reconstitution with the cofactor coelenterazine, emit
light when there are increases in free calcium concentrations
[Rizzuto R, Simpson A W, Brini M, Pozzan T, Nature 358, 325-327
(1992); Illarionov B A, Bondar V S, Illarionova V A, Vysotski E S,
Gene 153 (2), 273-274 (1995)]. The resulting vasopressin receptor
cells react to stimulation of the recombinantly expressed V1a
receptors by intracellular release of calcium ions, which can be
quantified by the resulting photoprotein luminescence. The
G.sub.s-coupled V2 receptors are stably transfected into cell lines
expressing the gene for firefly luciferase under control of a
CRE-responsible promoter. Activation of V2 receptors induces the
activation of the CRE-responsive promoter via cAMP increase,
thereby inducing the expression of firefly luciferase. The light
emitted by photoproteins of V1a cell lines as well as the light
emitted by firefly luciferase of V2 cell lines corresponds to the
activation or inhibition of the respective vasopressin receptor.
The bioluminescence of the cell lines is detected using a suitable
luminometer [Milligan G, Marshall F, Rees S, Trends in
Pharmacological Sciences 17, 235-237 (1996)].
Test Procedure:
Vasopressin V1a Receptor Cell Lines:
[0561] On the day before the assay, the cells are plated out in
culture medium (DMEM/F12, 2% FCS, 2 mM glutamine, 10 mM HEPES, 5
.mu.g/ml coelenterazine) in 384-well microtiter plates and kept in
a cell incubator (96% humidity, 5% v/v CO.sub.2, 37.degree. C.). On
the day of the assay, test compounds in various concentrations are
placed for 10 minutes in the wells of the microtiter plate before
the agonist [Arg.sup.8]-vasopressin at EC.sub.50 concentration is
added. The resulting light signal is measured immediately in the
luminometer.
Vasopressin V2 Receptor Cell Lines:
[0562] On the day before the assay, the cells are plated out in
culture medium (DMEM/F12, 2% FCS, 2 mM glutamine, 10 mM HEPES) in
384-well microtiter plates and kept in a cell incubator (96%
humidity, 5% v/v CO.sub.2, 37.degree. C.). On the day of the assay,
test compounds in various concentrations and the agonist
[Arg.sup.8]-vasopressin at EC.sub.50 concentration are added
together to the wells, and plates are incubated for 3 hours in a
cell incubator. Upon addition of the cell lysis reagent Triton.TM.
and the substrate luciferin, luminescence of firefly luciferase is
measured in a luminometer.
[0563] Table 1A below lists individual IC.sub.50 values for the
compounds of the invention (including diastereomeric mixtures as
well as separated, enantiopure diastereomers) that were obtained
from cell lines transfected with the human V1a or V2 receptor:
TABLE-US-00001 TABLE 1A Example IC.sub.50 hV1a IC.sub.50 hV2 No.
[.mu.M] [.mu.M] 1 0.0060 0.0025 2 0.0050 0.0087 3 0.0010 0.0056 4
0.0004 0.0053 5 0.0004 0.0018 6 0.0106 0.0017 7 0.0076 0.0026 8
0.0012 0.0107 9 0.0004 0.0023 10 0.0014 0.0014 11 0.0004 0.0003 12
0.0013 0.0011 13 0.0062 0.0014 14 0.0013 0.0004 15 0.0384 0.0041 16
0.0031 0.0060 17 0.0027 0.0034 18 0.0141 0.0086 19 0.0124 0.0014 20
0.0038 0.0008 21 0.0578 0.0022 22 0.0244 0.0023 23 0.0122 0.0009 24
0.1200 0.0020 25 0.0072 0.0036 26 0.0031 0.0030 27 0.0437 0.0077 28
0.0013 0.0002 29 0.0029 0.0002 30 0.0716 0.0004 31 0.0016 0.0009 32
0.0009 0.0010 33 0.0016 0.0023 34 0.0004 0.0012 35 0.0005 0.0016 36
0.0008 0.0028 37 0.0005 0.0007 38 0.0006 0.0009 39 0.0015 0.0035 40
0.0015 0.0072 41 0.0018 0.0079 42 0.0051 0.0127 43 0.0062 0.0012 44
0.0061 0.0012 45 0.0921 0.0033 46 0.0063 0.0021 47 0.0189 0.0037 48
0.0032 0.0024 49 0.0018 0.0126 50 0.0013 0.0100 51 0.0030 0.0223 52
0.0039 0.0004 53 0.0079 0.0018 54 0.0397 0.0016 55 0.0148 0.0042 56
0.0024 0.0014 57 0.0382 0.0082 58 0.0002 0.0015 59 0.0005 0.0024 60
0.0005 0.0052 61 0.0032 0.0002 62 0.0078 0.0009 63 0.0516 0.0019 64
0.0081 0.0051 65 0.0025 0.0033 66 0.0040 0.0019 67 0.0021 0.0027 68
0.0033 0.0013 70 0.0005 0.0011 71 0.0006 0.0021 73 0.0011 0.0070 74
0.0022 0.0247 75 0.0029 0.0066 76 0.0025 0.0051 77 0.0125 0.0135 78
0.0104 0.0031 79 0.0036 0.0017 80 0.0463 0.0051 82 0.0007 0.0023 83
0.0010 0.0067
[0564] The IC.sub.50 data listed in Table 1A demonstrate that the
compounds of the present invention are acting as highly potent dual
antagonists of vasopressin V1a and V2 receptors.
[0565] For comparative purposes, selected phenyl-triazole and
imidazole derivatives that were regarded to be representative of
closest prior art (cf. Int. Pat. Appl. WO 2011/104322-A1 and
example compounds described therein) were also tested in the
cellular V1a and V2 assays described above. IC.sub.50 values for
these compounds obtained from cell lines transfected with the human
V1a or V2 receptor are listed in Table 1B below:
TABLE-US-00002 TABLE 1B Example No. WO 2011/104322 IC.sub.50 hV1a
[.mu.M] IC.sub.50 hV2 [.mu.M] 54 0.0166 0.0564 56 0.0013 0.0067 60
0.0542 0.0326 68 0.0060 0.0083 101 0.0422 0.0238 110 0.0152
0.0043
B-2. Radioactive Binding Assay
[0566] IC.sub.50 and K.sub.i values were determined in a
radioactive binding competition SPA assay using membrane fractions
of recombinant CHO cell lines expressing the respective human, rat
or dog vasopressin V1a and V2 receptors. These cells derive from a
hamster's ovary epithelial cell (Chinese Hamster Ovary, CHO K1,
ATCC: American Type Culture Collection, Manassas, Va. 20108, USA).
In addition, the cells are stably transfected with the human, rat
or dog V1a or V2 receptor. The membrane preparations were subjected
to the radioactive receptor binding competition assay described
below.
[0567] The respective vasopressin receptor-transfected CHO cells
were grown in an appropriate quantity in T-175 flasks with
DMEM/F12, 10% FCS, 15 mM HEPES, 1 mg/ml G418 and kept in a cell
incubator (96% humidity, 5% v/v CO.sub.2, 37.degree. C.). After
reaching the appropriate confluency, cells were harvested for
membrane preparation. Cells were scraped into PBS and pelleted by
gentle centrifugation at 200.times.g for 5 min at room temperature.
Pellets were re-suspended in PBS and again centrifugated. After
repeating this step once more, the resulting pellets were
shock-frozen at -80.degree. C. for 30 min. Frozen pellets were
re-suspended in ice-cold preparation buffer (50 mM TRIS, 2 mM EDTA,
2 mM DTT, cOmplete Protease Inhibitor Cocktail) and homogenized at
2000 rpm for 35 seconds (Polytron PT3000, Kinematica). The
homogenate was cooled down for 2 min on ice, and the homogenization
was repeated twice. The resulting homogenate was centrifugated at
500.times.g for 10 min at 4.degree. C. Membranes were pelleted at
4500.times.g for 20 min at 4.degree. C., re-suspended in storing
buffer (7.5 mM TRIS, 12.5 mM MgCl.sub.2, 0.3 mM EDTA, 250 mM
sucrose, cOmplete Protease Inhibitor Cocktail) and homogenized at
2000 rpm for 2 seconds (Polytron PT3000, Kinematica). The protein
concentration was determined by using the BCA Protein Assay (Thermo
Scientific Pierce), and the membrane preparations were stored at
-80.degree. C. On the day of use, aliquots were thawed and briefly
vortexed.
[0568] For the determination of the receptor binding affinity of
test compounds, an SPA assay was set-up as follows. For each
membrane preparation, K.sub.d and B.sub.max values were determined.
From these data, the number of SPA beads (WGA PVT beads,
PerkinElmer, 200 .mu.g/well), the concentration of radioactive
ligand (.sup.3H-AVP, PerkinElmer, 2.431 TBq/mmol, f.c.
1-2.times.K.sub.d) and the amount of the respective membrane
preparation (10 .mu.g protein/well) were matched to the assay
volume (100 .mu.l) in binding buffer (50 mM TRIS, 0.2% BSA) in a
96-well plate. The test compounds were diluted in binding buffer
(f.c. 10.sup.-4 M to 10.sup.-12 M) and subjected to the assay.
Plates were gently shaken for 1-3 hours at room temperature and
further incubated for 1-2 hours. Signals generated by bound
.sup.3H-AVP were measured using a .beta.-counter (1450 Microbeta
Trilux). From these results, IC.sub.50 and K.sub.i values for the
tested compounds were calculated using GraphPad Prism.
B-3. Cellular In Vitro Assay for Detecting the Action of
Vasopressin V1a Receptor Antagonists on the Regulation of
Pro-Fibrotic Genes
[0569] The cell line H9C2 (American Type Culture Collection No.
CRL-1446), described as a cardiomyocyte type isolated from rat
cardiac tissue, endogenously expresses the vasopressin V1A receptor
AVPR1A in high copy number, whereas AVPR2 expression cannot be
detected. For cell assays for the inhibition of AVPR1A
receptor-dependent regulation of gene expression by receptor
antagonists, the procedure is as follows:
[0570] H9C2 cells are seeded in 6-well microtiter plates for cell
culture at a cell density of 50 000 cells/well in 2.0 ml of
Opti-MEM medium (Invitrogen Corp., Carlsbad, Calif., USA, Cat. No.
11058-021) and held in a cell incubator (96% humidity, 8% v/v
CO.sub.2, 37.degree. C.). After 24 hours, sets of three wells
(triplicate) are charged with vehicle solution (negative control)
and vasopressin solution ([Arg.sup.8]-vasopressin acetate, Sigma,
Cat. No. V9879), or test compound (dissolved in vehicle: water with
20% v/v ethanol) and vasopressin solution. In the cell culture, the
final vasopressin concentration is 1 nM. The test compound solution
is added to the cell culture in small volumes, so that a final
concentration of 0.03% of ethanol in the cell assay is not
exceeded. After an incubation time of 5 hours, the culture
supernatant is drawn off under suction, the adherent cells are
lysed in 350 .mu.l of RLT buffer (Qiagen, Cat. No. 79216), and the
RNA is isolated from the lysate using the RNeasy kit (Qiagen, Cat.
No. 74104). This is followed by DNAse digestion (Invitrogen, Cat.
No. 18068-015), cDNA synthesis (Promaga, ImProm-II Reverse
Transcription System, Cat. No. A3800) and RTPCR (pPCR MasterMix
RT-QP2X-03-075, Eurogentec, Seraing, Belgium). All procedures take
place in accordance with the working protocols of the test
reagents' manufacturers. The primer sets for the RTPCR are selected
on the basis of the mRNA gene sequences (NCBI GenBank Entrez
Nucleotide Data Base) using the Primer3Plus program with 6-FAM
TAMRA-labelled probes. The RTPCR for determining the relative mRNA
expression in the cells of the various assay batches is carried out
using the Applied Biosystems ABI Prism 7700 Sequence Detector in
384-well microtiter plate format in accordance with the instrument
operating instructions. The relative gene expression is represented
by the delta-delta Ct value [Applied Biosystems, User Bulletin No.
2 ABI Prism 7700 SDS, Dec. 11, 1997 (updated October 2001)] with
reference to the level of expression of the ribosomal protein L-32
gene (GenBank Acc. No. NM_013226) and the threshold Ct value of
Ct=35.
B-4. In Vivo Assay for Detecting Cardiovascular Effects: Blood
Pressure Measurement in Anaesthetized Rats (Vasopressin `Challenge`
Model)
[0571] In male Sprague-Dawley rats (250-350 g body weight) under
ketamine/xylazine/pentobarbital injection anaesthesia, polyethylene
tubes (PE-50, Intramedic.RTM.), which are prefilled with
heparin-containing (500 IU/ml) isotonic sodium chloride solution,
are introduced into the jugular vein and the femoral vein and then
tied in. Via one venous access, with the aid of a syringe,
Arg-vasopressin is injected; the test substance is administered via
the second venous access. For determination of the systolic blood
pressure, a pressure catheter (Millar SPR-320 2F) is tied into the
carotid artery. The arterial catheter is connected to a pressure
transducer which feeds its signals to a recording computer equipped
with suitable recording software. In a typical experiment, the
experimental animal is administered 3-4 successive bolus injections
at intervals of 10-15 min with a defined amount of Arg-vasopressin
(30 ng/kg) in isotonic sodium chloride solution. When the blood
pressure has reached initial levels again, the test substance is
administered as a bolus, with subsequent continuous infusion, in a
suitable solvent. After this, at defined intervals (10-15 min), the
same amount of Arg-vasopressin as at the start is administered
again. On the basis of the blood pressure values, a determination
is made of the extent to which the test substance counteracts the
hypertensive effect of Arg-vasopressin. Control animals only
receive solvent instead of the test substance.
[0572] Following intravenous administration, the compounds of the
invention, in comparison to the solvent controls, bring about an
inhibition of the blood pressure increase caused by
Arg-vasopressin.
B-5. In Vivo Assay for Detecting Cardiovascular Effects: Diuresis
Investigations in Conscious Rats Kept in Metabolism Cages
[0573] Wistar rats (220-450 g body weight) are kept with free
access to feed (Altromin) and drinking water. During the
experiment, the animals are kept with free access to drinking water
for 4 to 8 or up to 24 hours individually in metabolism cages
suitable for rats of this weight class (Tecniplast Deutschland
GmbH, D-82383 Hohenpei.beta.enberg). At the beginning of the
experiment, the animals are administered the test substance in a
volume of 1 to 3 ml/kg body weight of a suitable solvent by means
of gavage into the stomach. Control animals only receive solvent.
Controls and substance tests are carried out in parallel on the
same day. Control groups and substance-dose groups each consist of
4 to 8 animals. During the experiment, the urine excreted by the
animals is collected continuously in a receiver at the base of the
cage. The volume of urine per time unit is determined separately
for each animal, and the concentration of urinary electrolytes is
measured by standard methods of flame photometry. Before the
beginning of the experiment, the body weight of the individual
animals is determined.
[0574] Following oral administration, in comparison with the
solvent control applications, the compounds of the invention bring
about an increased excretion of urine, which is based essentially
on an increased excretion of water (aquaresis).
[0575] Table 2A below shows observed changes in urinary excretion
relative to solvent control (=100%) for exemplary compounds of the
invention at two different dosages:
TABLE-US-00003 TABLE 2A Urinary volume Dosage Urinary volume
Example Dosage p.o. [% vs. control = p.o. [% vs. control = No.
[mg/kg] 100%] [mg/kg] 100%] 1 0.3 194 3.0 588 2 0.3 194 3.0 588 3
0.3 89 3.0 450 4 0.3 91 1.0 190 5 0.3 166 1.0 438 6 0.3 132 1.0 439
11 0.3 159 3.0 443 12 0.3 96 3.0 323 14 -- -- 1.0 753 15 0.3 412
3.0 1085 17 0.3 404 1.0 819 29 0.3 404 1.0 983 70 0.3 139 3.0 595
76 0.3 350 3.0 1257 79 0.3 612 3.0 1312 82 0.3 220 3.0 828 83 0.3
279 3.0 1094
[0576] For comparative purposes, selected phenyl-triazole and
imidazole derivatives that were regarded to be representative of
closest prior art (cf. Int. Pat. Appl. WO 2011/104322-A1 and
example compounds described therein) were also tested for diuretic
effect in this assay. Observed changes in urinary excretion
relative to solvent control (=100%) at two different dosages are
shown in Table 2B below:
TABLE-US-00004 TABLE 2B Example No. Dosage Urinary volume Dosage
Urinary volume WO p.o. [% vs. control = p.o. [% vs. control =
2011/104322 [mg/kg] 100%] [mg/kg] 100%] 54 0.3 85 3.0 188 56 0.3
128 3.0 85 60 0.3 96 3.0 84 68 0.3 87 3.0 121 101 0.3 111 3.0 255
110 0.3 114 3.0 274
[0577] The results shown in Table 2A and 2B demonstrate that the
compounds of the present invention are significantly more potent in
vivo: Tested examples of the present invention gave rise to more
than a threefold, in some cases to more than a tenfold increase in
urinary volume versus the vehicle control group at a p.o. dose of 3
mg/kg, and most examples exhibited substantial aquaretic activity
already at p.o. doses of 0.3 mg/kg or 1 mg/kg. This is in contrast
to the phenyl-triazole and imidazole derivatives regarded to be
representative of closest prior art which were not active at p.o.
doses below 3 mg/kg and slightly active at 3 mg/kg.
B-6. In Vivo Assay for Detecting Cardiovascular Effects:
Hemodynamic Investigations in Anaesthetized Dogs
[0578] Male beagle dogs (Beagle, Marshall BioResources) with a
weight of between 10 and 15 kg are anaesthetized with pentobarbital
(30 mg/kg i.v., Narcoren.RTM., Merial, Germany) for the surgical
interventions and hemodynamic and functional examinations.
Pancuronium bromide (2 mg/animal i.v., Ratiopharm, Germany) serves
additionally as a muscle relaxant. The dogs are intubated and
ventilated with an oxygen/ambient air mixture (40/60%, about 3-4
L/min). Ventilation is carried out using a ventilator from GE
Healthcare (Avance) and is monitored using an analyzer
(Datex-Ohmeda, GE). Anaesthesia is maintained by continuous
infusion of pentobarbital (50 .mu.g/kg/min); fentanyl is used as an
analgesic (10-40 .mu.g/kg/h). An alternative to pentobarbital is to
use isoflurane (1-2% by volume).
[0579] In preparatory interventions, the dogs are fitted with a
cardiac pacemaker. At a time of 21 days before the first drug
testing (i.e. start of experiment), a cardiac pacemaker from
Biotronik (Logos.RTM.) is implanted into a subcutaneous skin pocket
and is contacted with the heart via a pacemaker electrode which is
advanced through the external jugular vein, with transillumination,
into the right ventricle. Thereafter all of the accesses are
removed, and the dog wakes spontaneously from the anaesthesia.
After a further 7 days (i.e. 14 days before the first drug
testing), the above-described pacemaker is activated, and the heart
is stimulated at a frequency of 220 beats per minute.
[0580] The actual substance testing experiments are carried out 14
and 28 days after the beginning of pacemaker stimulation, using the
following instrumentation: [0581] introduction of a bladder
catheter for bladder relief and for measuring the flow of urine;
[0582] attachment of ECG leads to the extremities for ECG
measurement; [0583] introduction of a Fluidmedic.RTM. PE 300 tube
filled with sodium chloride solution into the femoral artery; this
tube is connected to a pressure sensor (Braun Melsungen, Germany)
for measuring systemic blood pressure; [0584] introduction of a
Millar Tip catheter (type 350 PC, Millar Instruments, Houston, USA)
through the left atrium or through a port secured in the carotid
artery, for measuring cardiac hemodynamics; [0585] introduction of
a Swan-Ganz catheter (CCOmbo 7.5F, Edwards, Irvine, USA) via the
jugular vein into the pulmonary artery, for measuring cardiac
output, oxygen saturation, pulmonary arterial pressures and central
venous pressure; [0586] siting of a venous catheter in the cephalic
vein, for infusing pentobarbital, for liquid replacement and for
blood sampling (determination of plasma levels of the test
substance or of other clinical blood values); [0587] siting of a
venous catheter in the saphenous vein, for infusing fentanyl and
for administration of the test substance; [0588] continuous
infusion of vasopressin (Sigma, 4 mU/kg/min); test compounds are
then administered and evaluated at different dosages under this
vasopressin infusion.
[0589] The primary signals are amplified if necessary (ACQ 7700
amplifier, DataSciences Inc., Minneapolis, USA, or
Edwards-Vigilance-Monitor, Edwards, Irvine, USA) and subsequently
fed into the Ponemah system (DataSciences Inc., Minneapolis, USA)
for evaluation. The signals are recorded continuously throughout
the experimental period, and are further processed digitally by the
software and averaged over 30 seconds.
[0590] Although the invention has been disclosed with reference to
specific embodiments, it is apparent that other embodiments and
variations of the invention may be devised by others skilled in the
art without departing from the true spirit and scope of the
invention. The claims are intended to be construed to include all
such embodiments and equivalent variations.
C. EXAMPLES RELATING TO PHARMACEUTICAL COMPOSITIONS
[0591] Pharmaceutical compositions according to the present
invention can be illustrated as follows:
Sterile i.v. Solution:
[0592] A 5 mg/mL solution of the desired compound of the invention
can be made using sterile, injectable water, and the pH is adjusted
if necessary. The solution is diluted for administration to 1-2
mg/mL with sterile 5% dextrose and is administered as an i.v.
infusion over about 60 minutes.
Lyophilized Powder for i.v. Administration:
[0593] A sterile preparation can be prepared with (i) 100-1000 mg
of the desired compound of the invention as a lyophilized powder,
(ii) 32-327 mg/mL sodium citrate, and (iii) 300-3000 mg Dextran 40.
The formulation is reconstituted with sterile, injectable saline or
5% dextrose to a concentration of 10 to 20 mg/mL, which is further
diluted with saline or 5% dextrose to 0.2 to 0.4 mg/mL, and is
administered either as i.v. bolus or by i.v. infusion over 15-60
minutes.
Intramuscular Suspension:
[0594] The following solution or suspension can be prepared for
intramuscular injection:
[0595] 50 mg/mL of the desired, water-insoluble compound of the
invention; 5 mg/mL sodium carboxymethylcellulose; 4 mg/mL Tween 80;
9 mg/mL sodium chloride; 9 mg/mL benzyl alcohol.
Hard Shell Capsules:
[0596] A large number of unit capsules are prepared by filling
standard two-piece hard gelatin capsules each with 100 mg of the
desired, powdered compound of the invention, 150 mg of lactose, 50
mg of cellulose and 6 mg of magnesium stearate.
Soft Gelatin Capsules:
[0597] A mixture of the desired compound of the invention in a
digestible oil, such as soybean oil, cottonseed oil or olive oil,
is prepared and injected by means of a positive displacement pump
into molten gelatin to form soft gelatin capsules containing 100 mg
of the active ingredient. The capsules are washed and dried. The
desired compound of the invention can be dissolved in a mixture of
polyethylene glycol, glycerin and sorbitol to prepare a
water-miscible medicine mix.
Tablets:
[0598] A large number of tablets are prepared by conventional
procedures so that the dosage unit is 100 mg of the desired
compound of the invention, 0.2 mg of colloidal silicon dioxide, 5
mg of magnesium stearate, 275 mg of microcrystalline cellulose, 11
mg of starch, and 98.8 mg of lactose. Appropriate aqueous and
non-aqueous coatings may be applied to increase palatability,
improve elegance and stability, or delay absorption.
* * * * *