HK1240931B - Hydroxyalkyl-substituted phenyltriazole derivatives and uses thereof - Google Patents

Description

Hydroxyalkyl substituted phenyltriazole derivatives and uses thereof

The present invention relates to novel 5-(hydroxyalkyl)-1-phenyl-1,2,4-triazole derivatives, methods for preparing the compounds, pharmaceutical compositions containing the compounds, and the use of the compounds or compositions for treating and/or preventing diseases, in particular for treating and/or preventing cardiovascular diseases and kidney diseases.

The body's fluid content is determined by various physiological control mechanisms, the goal of which is to maintain a constant fluid level (volume homeostasis). In this process, the volumetric filling of the vascular system and the osmotic pressure of the plasma are continuously recorded by appropriate receptors (baroreceptors and osmoreceptors). These receptors provide information to relevant brain centers, which regulate drinking behavior and control fluid excretion via the kidneys through humoral and neural signals. The peptide hormone vasopressin plays a key role in this process [Schrier RW, Abraham WT, New Engl. J. Med. 341 , 577-585 (1999)].

Vasopressin is produced in specialized endocrine neurons in the supraoptic and paraventricular nuclei in the walls of the third ventricle (hypothalamus). From there, it is transported along the neurites of these neurons to the posterior pituitary gland (neurohypophysis). There, the hormone is released into the bloodstream in response to stimuli. Volume loss (e.g., due to acute hemorrhage, heavy sweating, prolonged thirst, or diarrhea) stimulates the release of this hormone in large quantities. Conversely, an increase in intravascular volume (e.g., due to increased fluid intake) inhibits vasopressin secretion.

Vasopressin exerts its effects primarily by binding to three receptors, classified as V1a, V1b, and V2, which belong to the G protein-coupled receptor family. V1a receptors are primarily located on cells of vascular smooth muscle tissue. Their activation leads to vasoconstriction, thereby increasing peripheral resistance and blood pressure. V1a receptors can also be detected in the liver. V1b receptors (also known as V3 receptors) can be detected in the central nervous system. Along with corticotropin-releasing hormone (CRH), vasopressin regulates basal and stress-induced secretion of adrenocorticotropic hormone (ACTH) via V1b receptors. V2 receptors are located in the epithelium of distal tubular epithelium and the renal collecting ducts. Their activation renders these epithelia permeable to water. This phenomenon is due to the presence of aquaporins (specialized water channels) embedded in the luminal membrane of epithelial cells.

The importance of vasopressin for reabsorbing water from the urine in the kidneys becomes clear from the clinical manifestations of diabetes insipidus, caused by hormone deficiency (e.g., due to pituitary gland damage). Without hormone replacement, patients suffering from this condition excrete up to 20 liters of urine per 24 hours. This volume corresponds to approximately 10% of the primary urine. Due to its crucial role in reabsorbing water from urine, vasopressin is also synonymously known as antidiuretic hormone (ADH). Therefore, pharmacological inhibition of the action of vasopressin/ADH at the V2 receptor leads to increased urinary excretion. However, in contrast to the effects of other diuretics (thiazides and loop diuretics), V2 receptor antagonists cause increased water excretion without significantly increasing electrolyte excretion. This means that V2 antagonists can restore volume homeostasis without affecting electrolyte homeostasis. Therefore, drugs with V2 antagonistic activity appear particularly suitable for treating all disease states associated with water overload without a simultaneous increase in electrolytes.

In clinical chemistry, significant electrolyte abnormalities can be measured as hyponatremia (sodium concentration < 135 mmol/L); it is the most important electrolyte abnormality in hospitalized patients, with an incidence of approximately 5% or 250,000 cases per year in the United States alone. If the plasma sodium concentration falls below 115 mmol/L, coma and death are imminent. Depending on the underlying cause, hypovolemic, euvolemic, and hypervolemic hyponatremia are distinguished. Clinically, the most important form of hypervolemia is that with edema formation. Typical examples include the syndrome of inappropriate ADH/vasopressin secretion (SIADH) (e.g., after head injury or as a symptom associated with cancer) and hypervolemic hyponatremia in cirrhosis, various renal diseases, and heart failure [DeLuca L. et al., Am. J. Cardiol. 96 (Suppl.), 19L-23L (2005)]. In particular, patients with heart failure, despite their relative hyponatremia and hypervolemia, often exhibit elevated vasopressin levels, which is considered a consequence of the pervasive perturbation of neurohumoral regulation in heart failure [Francis GS et al., Circulation 82 , 1724-1729 (1990)].

Disrupted neurohumoral regulation primarily manifests as increased sympathetic tone and inappropriate activation of the renin-angiotensin-aldosterone system. Currently, inhibition of these components through β-blockers and ACE inhibitors or angiotensin receptor blockers is an integral component of pharmacological treatment for heart failure. However, inadequate treatment of inappropriately elevated vasopressin secretion in advanced heart failure remains inadequate. In addition to water retention mediated by V2 receptors and the associated adverse hemodynamic consequences of increased afterload, left ventricular emptying, pulmonary vascular pressure, and cardiac output are also adversely affected by V1a-mediated vasoconstriction. Furthermore, based on animal data, a direct hypertrophic effect on the myocardium has been attributed to vasopressin. Unlike the renal volume expansion effect, which is mediated by V2 receptor activation, this direct effect on the myocardium is triggered by activation of V1a receptors.

For these reasons, agents that inhibit the effects of vasopressin on V2 and/or V1a receptors appear to be suitable for the treatment of heart failure. In particular, compounds with combined activity at both vasopressin receptors (V1a and V2) would have desirable renal and hemodynamic effects and thus offer a particularly desirable profile for treating patients with heart failure. Providing such combined vasopressin antagonists also seems reasonable, as volume reduction mediated solely by V2 receptor blockade could lead to stimulation of osmoreceptors and, therefore, a further compensatory increase in vasopressin release. Consequently, in the absence of a component that also blocks V1a receptors, the deleterious effects of vasopressin, such as vasoconstriction and myocardial hypertrophy, could be further potentiated [Saghi P. et al., Europ. Heart J. 26 , 538-543 (2005)].

Certain 4-phenyl-1,2,4-triazol-3-yl derivatives have been described in WO 2005/063754-A1 and WO 2005/105779-A1 as vasopressin V1a receptor antagonists which can be used for the treatment of gynecological disorders, in particular menstrual disorders such as dysmenorrhea.

WO 2011/104322-A1 discloses a specific group of bisaryl-linked 1,2,4-triazol-3-ones (including their 5-phenyl-1,2,4-triazol-3-yl and 1-phenyl-1,2,3-triazol-4-yl derivatives) as vasopressin V1a and/or V2 receptor antagonists useful for the treatment and/or prevention of cardiovascular disease. However, during further investigation of this structural class, candidate compounds were often hampered by unsatisfactory water-stimulating potency when evaluated in vivo following oral administration to conscious rats. However, as noted above, robust water-stimulating efficacy is a desirable prerequisite for treating conditions associated with water overload, such as congestive heart failure.

A significant increase in water-stimulating potency will also help reduce the amount of substance required to achieve and maintain the desired therapeutic effect, thereby limiting the possibility of unacceptable side effects and/or unwanted drug-drug interactions in treating patients who may already be at high risk for, for example, acute or chronic heart failure or renal failure.

The technical problem underlying the present invention can therefore be seen as the identification and provision of novel compounds that act as potent vasopressin V1a and V2 receptor antagonists and that additionally exhibit a significantly increased water-inducing capacity in vivo.

Surprisingly, it has now been discovered that certain 5-(hydroxyalkyl)-1-phenyl-1,2,4-triazole derivatives are very potent vasopressin V1a and V2 receptor antagonists which, after oral administration, exhibit a significantly enhanced water-promoting effect in vivo. This enhanced activity profile makes the compounds of the invention particularly useful for the treatment and/or prevention of cardiovascular and renal diseases.

In one aspect, the present invention relates to 5-(hydroxyalkyl)-1-phenyl-1,2,4-triazole derivatives of the general formula (I),

in

^R1 is hydrogen or methyl, and

R ^2A and R ^2B are each independently selected from hydrogen, fluoro, chloro, cyano, methyl, fluoromethyl, difluoromethyl, trifluoromethyl, ethyl, methoxy, difluoromethoxy, and trifluoromethoxy.

The compounds according to the invention may also be present in the form of their salts, solvates and/or solvates of the salts.

The compounds according to the present invention are compounds of formula (I) and their salts, solvates and solvates of said salts; compounds of formula (I) contained in the chemical formula mentioned below and their salts, solvates and solvates of said salts; and compounds contained in formula (I) and mentioned below as process products and/or embodiment examples and their salts, solvates and solvates of said salts; compounds contained in formula (I) and mentioned below that have not yet become salts, solvates and solvates of said salts.

The salts used in the present invention are preferably pharmaceutically acceptable salts of the compounds according to the invention (see, for example, SM Berge et al., "Pharmaceutical Salts", J. Pharm. Sci. 1977, 66, 1-19). These also include salts which, although not themselves suitable for pharmaceutical use, can be used, for example, to isolate, purify or store the compounds according to the invention.

Pharmaceutically acceptable salts include acid addition salts of inorganic acids, carboxylic acids, and sulfonic acids, such as hydrochloric acid, hydrobromic acid, sulfuric acid, phosphoric acid, methanesulfonic acid, ethanesulfonic acid, benzenesulfonic acid, toluenesulfonic acid, naphthalene disulfonic acid, formic acid, acetic acid, trifluoroacetic acid, propionic acid, lactic acid, tartaric acid, malic acid, citric acid, fumaric acid, maleic acid, and benzoic acid.

Pharmaceutically acceptable salts also include salts with conventional bases, such as alkali metal salts (e.g., sodium and potassium salts), alkaline earth metal salts (e.g., calcium and magnesium salts), and ammonium salts derived from ammonia or organic amines, such as, for example 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.

For the purposes of the present invention, solvates are those forms of the compounds according to the invention which form complexes in the solid or liquid state by stoichiometric coordination with solvent molecules. Hydrates are a specific form of solvates, in which the coordination occurs with water. Hydrates are the preferred solvates for the purposes of the present invention.

The compounds of the present invention may exist as isomers (enantiomers, diastereomers) due to asymmetric centers or due to hindered rotation. Any isomer may exist in which the asymmetric center is in ( R )-, ( S )- or ( R,S ) configuration.

It will also be appreciated that when two or more asymmetric centers are present in the compounds of the present invention, multiple diastereomers and enantiomers of the exemplified structures are often possible, and that pure diastereomers and pure enantiomers represent preferred embodiments. This means that pure stereoisomers, pure diastereomers, pure enantiomers, and mixtures thereof are all within the scope of the present invention.

All isomers of the compounds of the present invention, whether isolated, pure, partially purified or as racemic mixtures, are included within the scope of the present invention. The purification of the isomers and the separation of the isomer mixtures can be achieved by standard techniques known in the art. For example, diastereomeric mixtures can be separated into individual isomers by chromatography or crystallization, and racemates can be separated into individual enantiomers by chromatography on a chiral phase or by resolution.

Furthermore, all possible tautomeric forms of the above compounds are also included in the present invention.

The present invention also encompasses all suitable isotopic variations of the compounds according to the invention. Isotopic variations of the compounds according to the invention are understood herein to mean compounds in which at least one atom in the compounds according to the invention has been replaced by another atom of the same atomic number, but the atomic mass of the other atom differs from the atomic mass normally or predominantly found in nature. Examples of isotopes that can be incorporated into the compounds according to the invention are isotopes of hydrogen, carbon, nitrogen, oxygen, fluorine, chlorine, bromine, and iodine, such as ^2H (deuterium), ^3H (tritium), ^13C , ^14C , ^15N , ^17O , ^18O , ^18F , ^36Cl , ^82Br , 123I, ^124I , ^129I , and ^{131I .} Certain isotopic variations of the compounds according to the invention, particularly those into which one or more radioactive isotopes have been incorporated, may be useful, for example, for examining the mechanism of action or the distribution of the active compound in the body. Due to the relatively easy preparability and detectability, special compounds labeled with ³ H, ¹⁴ C and/or ¹⁸ F isotopes are suitable for this purpose. In addition, due to the greater metabolic stability of the compound, the incorporation of an isotope (e.g., deuterium) can lead to specific therapeutic benefits, such as an extension of the half-life in vivo or a reduction in the required active dose. Therefore, such modifications of the compounds according to the present invention may also constitute preferred embodiments of the present invention in some cases. Isotopic variants of the compounds according to the present invention can be prepared by methods known to those skilled in the art, such as by the methods described below and in the working examples, by using corresponding isotopic modifications of specific reagents and/or starting compounds therein.

In a particular embodiment, the invention relates to compounds of formula (I), wherein R ¹ is methyl.

In another embodiment, the invention relates to compounds of formula (I), wherein at least one of R ^2A and R ^2B is not hydrogen.

In yet another embodiment, the invention relates to compounds of formula (I), wherein R ¹ is hydrogen or methyl, and

R ^2A and R ^2B are independently selected from hydrogen, fluorine, chlorine, methyl and methoxy, wherein at least one of R ^2A and R ^2B is not hydrogen.

In a preferred embodiment, the present invention relates to compounds according to formula (I) selected from the group consisting of:

5-(4-chlorophenyl)-2-{[1-(3-chlorophenyl)-5-(hydroxymethyl)-1 H -1,2,4-triazol-3-yl]methyl}-4-[(2 S )-3,3,3-trifluoro-2-hydroxypropyl]-2,4-dihydro-3 H -1,2,4-triazol-3-one;

5-(4-chlorophenyl)-2-{[1-(3-fluorophenyl)-5-(hydroxymethyl)-1 H -1,2,4-triazol-3-yl]methyl}-4-[(2 S )-3,3,3-trifluoro-2-hydroxypropyl]-2,4-dihydro-3 H -1,2,4-triazol-3-one;

5-(4-chlorophenyl)-2-{[5-(hydroxymethyl)-1-(2-methylphenyl)-1 H -1,2,4-triazol-3-yl]methyl}-4-[(2 S )-3,3,3-trifluoro-2-hydroxypropyl]-2,4-dihydro-3 H -1,2,4-triazol-3-one;

2-({1-(2-chloro-4-fluorophenyl)-5-[(1 RS )-1-hydroxyethyl]-1 H -1,2,4-triazol-3-yl}methyl)-5-(4-chlorophenyl)-4-[(2 S )-3,3,3-trifluoro-2-hydroxypropyl]-2,4-dihydro-3 H -1,2,4-triazol-3-one;

2-{[1-(2-chloro-4-fluorophenyl)-5-(1-hydroxyethyl)-1 H -1,2,4-triazol-3-yl]methyl}-5-(4-chlorophenyl)-4-[(2 S )-3,3,3-trifluoro-2-hydroxypropyl]-2,4-dihydro-3 H -1,2,4-triazol-3-one ( diastereomer 1 );

2-{[1-(2-chloro-4-fluorophenyl)-5-(1-hydroxyethyl)-1 H -1,2,4-triazol-3-yl]methyl}-5-(4-chlorophenyl)-4-[(2 S )-3,3,3-trifluoro-2-hydroxypropyl]-2,4-dihydro-3 H -1,2,4-triazol-3-one ( diastereomer 2 );

2-({1-(2-chloro-5-fluorophenyl)-5-[(1 RS )-1-hydroxyethyl]-1 H -1,2,4-triazol-3-yl}methyl)-5-(4-chlorophenyl)-4-[(2 S )-3,3,3-trifluoro-2-hydroxypropyl]-2,4-dihydro-3 H -1,2,4-triazol-3-one;

2-{[1-(2-chloro-5-fluorophenyl)-5-(1-hydroxyethyl)-1 H -1,2,4-triazol-3-yl]methyl}-5-(4-chlorophenyl)-4-[(2 S )-3,3,3-trifluoro-2-hydroxypropyl]-2,4-dihydro-3 H -1,2,4-triazol-3-one ( diastereomer 1 );

2-{[1-(2-chloro-5-fluorophenyl)-5-(1-hydroxyethyl)-1 H -1,2,4-triazol-3-yl]methyl}-5-(4-chlorophenyl)-4-[(2 S )-3,3,3-trifluoro-2-hydroxypropyl]-2,4-dihydro-3 H -1,2,4-triazol-3-one ( diastereomer 2 );

5-(4-chlorophenyl)-2-({1-(3-fluorophenyl)-5-[(1 RS )-1-hydroxyethyl]-1 H -1,2,4-triazol-3-yl}methyl)-4-[(2 S )-3,3,3-trifluoro-2-hydroxypropyl]-2,4-dihydro-3 H -1,2,4-triazol-3-one;

5-(4-chlorophenyl)-2-({1-(3-fluorophenyl)-5-[(1 R )-1-hydroxyethyl]-1 H -1,2,4-triazol-3-yl}methyl)-4-[(2 S )-3,3,3-trifluoro-2-hydroxypropyl]-2,4-dihydro-3 H -1,2,4-triazol-3-one;

5-(4-chlorophenyl)-2-({1-(3-fluorophenyl)-5-[(1 S )-1-hydroxyethyl]-1 H -1,2,4-triazol-3-yl}methyl)-4-[(2 S )-3,3,3-trifluoro-2-hydroxypropyl]-2,4-dihydro-3 H -1,2,4-triazol-3-one;

5-(4-chlorophenyl)-2-({1-(3-chlorophenyl)-5-[(1 RS )-1-hydroxyethyl]-1 H -1,2,4-triazol-3-yl}methyl)-4-[(2 S )-3,3,3-trifluoro-2-hydroxypropyl]-2,4-dihydro-3 H -1,2,4-triazol-3-one;

5-(4-chlorophenyl)-2-({1-(3-chlorophenyl)-5-[(1 R )-1-hydroxyethyl]-1 H -1,2,4-triazol-3-yl}methyl)-4-[(2 S )-3,3,3-trifluoro-2-hydroxypropyl]-2,4-dihydro-3 H -1,2,4-triazol-3-one;

5-(4-chlorophenyl)-2-({1-(3-chlorophenyl)-5-[(1 S )-1-hydroxyethyl]-1 H -1,2,4-triazol-3-yl}methyl)-4-[(2 S )-3,3,3-trifluoro-2-hydroxypropyl]-2,4-dihydro-3 H -1,2,4-triazol-3-one;

5-(4-chlorophenyl)-2-({1-(2-chlorophenyl)-5-[(1 RS )-1-hydroxyethyl]-1 H -1,2,4-triazol-3-yl}methyl)-4-[(2 S )-3,3,3-trifluoro-2-hydroxypropyl]-2,4-dihydro-3 H -1,2,4-triazol-3-one;

5-(4-chlorophenyl)-2-({1-(2-chlorophenyl)-5-[(1 R )-1-hydroxyethyl]-1 H -1,2,4-triazol-3-yl}methyl)-4-[(2 S )-3,3,3-trifluoro-2-hydroxypropyl]-2,4-dihydro-3 H -1,2,4-triazol-3-one; and

5-(4-chlorophenyl)-2-({1-(2-chlorophenyl)-5-[(1 S )-1-hydroxyethyl]-1 H -1,2,4-triazol-3-yl}methyl)-4-[(2 S )-3,3,3-trifluoro-2-hydroxypropyl]-2,4-dihydro-3 H -1,2,4-triazol-3-one.

In a particularly preferred embodiment, the present invention relates to compounds according to formula (I) selected from the group consisting of:

5-(4-chlorophenyl)-2-({1-(3-fluorophenyl)-5-[(1 RS )-1-hydroxyethyl]-1 H -1,2,4-triazol-3-yl}methyl)-4-[(2 S )-3,3,3-trifluoro-2-hydroxypropyl]-2,4-dihydro-3 H -1,2,4-triazol-3-one;

5-(4-chlorophenyl)-2-({1-(3-fluorophenyl)-5-[(1 S )-1-hydroxyethyl]-1 H -1,2,4-triazol-3-yl}methyl)-4-[(2 S )-3,3,3-trifluoro-2-hydroxypropyl]-2,4-dihydro-3 H -1,2,4-triazol-3-one;

5-(4-chlorophenyl)-2-({1-(3-chlorophenyl)-5-[(1 RS )-1-hydroxyethyl]-1 H -1,2,4-triazol-3-yl}methyl)-4-[(2 S )-3,3,3-trifluoro-2-hydroxypropyl]-2,4-dihydro-3 H -1,2,4-triazol-3-one;

5-(4-chlorophenyl)-2-({1-(3-chlorophenyl)-5-[(1 S )-1-hydroxyethyl]-1 H -1,2,4-triazol-3-yl}methyl)-4-[(2 S )-3,3,3-trifluoro-2-hydroxypropyl]-2,4-dihydro-3 H -1,2,4-triazol-3-one;

5-(4-chlorophenyl)-2-({1-(2-chlorophenyl)-5-[(1 RS )-1-hydroxyethyl]-1 H -1,2,4-triazol-3-yl}methyl)-4-[(2 S )-3,3,3-trifluoro-2-hydroxypropyl]-2,4-dihydro-3 H -1,2,4-triazol-3-one; and

5-(4-chlorophenyl)-2-({1-(2-chlorophenyl)-5-[(1 S )-1-hydroxyethyl]-1 H -1,2,4-triazol-3-yl}methyl)-4-[(2 S )-3,3,3-trifluoro-2-hydroxypropyl]-2,4-dihydro-3 H -1,2,4-triazol-3-one.

In another embodiment, the present invention relates to a process for preparing a compound of formula (I), characterized in that firstly a compound of formula (II) is

Reaction with hydrazine to obtain hydrazide of formula (III),

Then, in the presence of a base, condense with an amidine of formula (IV) or a salt thereof

wherein R ¹ has the above meaning,

To obtain a 1,2,4-triazole derivative of formula (V) and/or its tautomer,

wherein R ¹ has the above meaning,

Then, in the presence of a copper catalyst and an amine base, the phenylboronic acid of formula (VI) is coupled

wherein R ^2A and R ^2B have the above-mentioned meanings,

And obtain the target compound of formula (I),

wherein R ¹ , R ^2A and R ^2B have the above-mentioned meanings,

Optionally, the compounds of formula (I) are then separated, where appropriate, by ( i ) preferably using chromatographic methods to thereby obtain their respective diastereomers, and/or ( ii ) converted into their respective hydrates, solvates, salts and/or hydrates or solvates of said salts by treatment with corresponding solvents and/or acids or bases.

Compounds of formula (I) (wherein R ¹ represents methyl) can also be obtained in diastereomerically pure form by using the appropriate enantiomer of amidine (IV) [R ¹ = methyl] (i.e. (IV-A) or (IV-B)) or a salt thereof in the above condensation reaction.

The conversion of (II)→(III) is carried out in a customary manner by treating the methyl ester (II) with hydrazine or hydrazine hydrate in an alcoholic solvent such as methanol, ethanol, n-propanol, isopropanol or n-butanol at temperatures from +20°C to +100°C.

The condensation reaction of (III) + (IV) → (V) is usually carried out in an inert dipolar aprotic solvent in the presence of a sufficiently strong base, such as N,N -dimethylformamide (DMF), N,N -dimethylacetamide (DMA), dimethyl sulfoxide (DMSO), N-methylpyrrolidone (NMP) or N ,N' -dimethylpropyleneurea (DMPU), wherein the sufficiently strong base is, for example, sodium hydride or a sodium or potassium alkoxide, such as sodium methoxide or potassium methoxide, sodium ethoxide or potassium ethoxide, or sodium tert-butoxide or potassium tert-butoxide. The amidine (IV) can be used in the reaction as is or in the form of a salt, such as a hydrochloride. In the latter case, a proportional excess of base is used. The reaction is usually carried out at a temperature of +80°C to +150°C. Heating with a microwave reactor apparatus may have a beneficial effect on the condensation reaction.

The 1,2,4-triazole derivative of formula (V) prepared by this reaction may also exist in other tautomeric forms, such as (V-A) or (V-B), or in the form of a tautomeric mixture.

The coupling reaction of (V) + (VI) → (I) is usually carried out with the aid of a copper catalyst and an amine base ["Chan-Lam coupling"conditions; see, for example, DMT Chan et al., Tetrahedron Lett. 44 (19), 3863-3865 (2003); JX Qiao and PYS Lam, Synthesis , 829-856 (2011); KS Rao and T.-S. Wu, Tetrahedron 68 , 7735-7754 (2012)]. Suitable copper catalysts for this process are, in particular, copper(II) salts, such as copper(II) acetate, copper(II) trifluoromethanesulfonate or copper(II) bromide. Useful amine bases include, for example, triethylamine, N,N -diisopropylethylamine, pyridine and 4-( N,N -dimethylamino)pyridine. The reaction is carried out 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 as the solvent and base. The coupling reaction is generally carried out at a temperature of +20°C to +120°C, preferably at a temperature of +20°C to +70°C. Accompanying microwave irradiation may also have a beneficial effect in this reaction.

If it occurs, the regioisomeric phenyltriazole derivatives [ cf. tautomers (VA), (VB)] resulting from the coupling reaction at the other triazole nitrogen atom can be easily separated from the target product (I) by conventional HPLC chromatography.

The compound of formula (II) can be synthesized by the procedure described in international patent application WO 2011/104322-A1 (see also Synthesis Schemes 1a and 1b below).

Compounds of formula (IV), (IV-A), (IV-B) and (VI) are commercially available, known from the literature, or can be prepared from readily available starting materials by adapting standard methods described in the literature. Detailed procedures and references for the preparation of the starting materials can also be found in the experimental section of the chapters on the preparation of starting materials and intermediates.

The preparation of the compounds of the present invention can be illustrated by the following synthetic scheme:

Option 1a

[ cf. International patent application WO 2011/104322-A1].

Option 1b

[ cf. International patent application WO 2011/104322-A1].

Option 2

.

The compounds of the present invention possess valuable pharmacological properties and can be used to prevent and/or treat a variety of diseases and disease-causing conditions in humans and other mammals.

In the context of the present invention, the terms "treatment" or "treating" include inhibiting, delaying, alleviating, relieving, preventing, reducing, or reversing a disease, disorder, condition, or state, its development and/or progression, and/or its symptoms. The terms "prevention" or "preventing" include reducing the risk of acquiring, contracting, or experiencing a disease, disorder, condition, or state, its development and/or progression, and/or its symptoms. The term prevention includes both prevention and treatment. Treatment or prevention of a disease, disorder, condition, or state may be partial or complete.

Throughout this document, for simplicity, singular language is preferred to plural language, but plural language is generally intended to be included unless otherwise indicated. For example, the expression "a method of treating a disease in a patient comprising administering to the patient an effective amount of a compound of formula (I)" is intended to include treating more than one disease simultaneously and administering more than one compound of formula (I).

The compounds of the present invention are highly potent dual antagonists of both vasopressin V1a and V2 receptors. Furthermore, following oral administration, the compounds of the present invention exhibit significant diuretic effects in vivo. Therefore, the compounds of the present invention are expected to be valuable as therapeutic agents for the treatment and/or prevention of diseases, particularly cardiovascular and renal diseases.

Cardiovascular diseases in the context of which the compounds of the present invention may be used to treat and/or prevent 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, pulmonary hypertension, coronary heart disease, stable and unstable angina, atrial and ventricular arrhythmias, atrial and ventricular rhythm disturbances and conduction disorders such as atrioventricular block (AVB) of grades I-III. I-III), supraventricular tachycardia, atrial fibrillation, atrial flutter, ventricular fibrillation, ventricular flutter, ventricular tachycardia, torsades de pointes, atrial and ventricular extrasystoles, AV nodal extrasystoles, sick sinus syndrome, syncope, AV-nodal reentrant tachycardia and Wolff-White syndrome, acute coronary syndrome (ACS), autoimmune heart diseases (pericarditis, endocarditis, valvulitis, aortitis, cardiomyopathy), shock such as cardiogenic shock, septic shock and anaphylactic shock, aneurysm, Boxer cardiomyopathy (ventricular premature beats), and thromboembolic diseases and ischemia such as peripheral perfusion disorders, reperfusion injury, arterial and venous thrombosis, myocardial insufficiency, endothelial dysfunction, microvascular and macrovascular injury (vasculitis) ), prevention of restenosis (e.g. after thrombolytic therapy, percutaneous transluminal angioplasty (PTA), percutaneous transluminal coronary angioplasty (PTCA), heart transplantation and bypass surgery), arteriosclerosis, lipid metabolism disorders, hypolipoproteinaemias, dyslipidemias, hypertriglyceridemias, hyperlipidemias and combined hyperlipidemias, hypercholesterolemias, abetalipoproteinemias, sitosterolemias, xanthomatosis, Tangier disease, excess fat, obesity, metabolic syndrome, transient ischemic attack, stroke, inflammatory cardiovascular disease, peripheral and heart vascular disease, peripheral circulatory disease, coronary and peripheral artery spasm, as well as edemas such as pulmonary edema, cerebral edema, renal edema and edema associated with heart failure.

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, total failure, ischemic cardiomyopathy, dilated cardiomyopathy, congenital heart defects, heart valve defects, heart failure associated with heart valve defects, mitral stenosis, mitral insufficiency, aortic stenosis, aortic insufficiency, tricuspid stenosis, tricuspid insufficiency, pulmonary stenosis, pulmonary insufficiency, complex heart valve defects, inflammation of the myocardium (myocarditis), chronic myocarditis, acute myocarditis, viral myocarditis, diabetic heart failure, alcoholic 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).

The compounds according to the invention are also suitable for the treatment and/or prevention of renal diseases, in particular acute and chronic renal insufficiency and acute and chronic renal failure. In the sense of the present invention, the term renal insufficiency includes acute and chronic clinical manifestations of renal insufficiency, as well as underlying or associated renal diseases, such as renal hypoperfusion, dialysis-related hypotension, urinary tract obstruction, glomerulopathies, glomerulonephritis, acute glomerulonephritis, glomerulosclerosis, tubulointerstitial diseases, renal diseases, such as primary and congenital renal diseases, nephritis, immune renal diseases, such as renal transplant rejection, immune complex-induced renal disease, toxic substance-induced nephropathy, contrast agent-induced nephropathy , diabetic and non-diabetic nephropathy, pyelonephritis, renal cysts, nephrosclerosis, hypertensive nephrosclerosis and nephrotic syndrome, which can be characterized in diagnosis by abnormally reduced creatinine and/or water excretion, abnormally increased blood concentrations of urea, nitrogen, potassium and/or creatinine, changed renal enzymes such as the activity of glutamine synthetase, changed urine osmotic pressure or urine volume, increased microalbuminuria, macroalbuminuria, lesions of glomeruli and arterioles, renal tubular dilatation, hyperphosphatemia and/or the need for dialysis. The present invention also includes the use of the compounds according to the invention for the treatment and/or prevention of sequelae of renal insufficiency, such as pulmonary edema, heart failure, uremia, anemia, electrolyte imbalances (such as hyperkalemia, hyponatremia) and bone and carbohydrate metabolism disorders.

The compounds of the present invention are particularly useful for treating and/or preventing cardiorenal syndrome (CRS) and its various subtypes. The term includes specific heart and kidney diseases, in which acute or chronic dysfunction of one organ may lead to acute or chronic dysfunction of another organ. CRS is subdivided into 5 types according to the organ that initiates the damage and the acuteness and chronicity of the disease (type Type 1: the development of renal insufficiency caused by acute decompensated heart failure; Type 2: caused by progressive renal dysfunction Chronic congestive heart failure; Type 3: acute cardiac insufficiency caused by a sudden decline in renal function; Type 4: Chronic kidney disease leads to cardiac remodeling; Type 5: systemic disease including heart and kidney) [see, for example, M.R.Kahn et al., Nature Rev.Cardiol.10, 261-273 (2013)].

Compound according to the present invention is also suitable for treating and/or preventing polycystic kidney disease (PCKD) and syndrome of inappropriate ADH secretion (SIADH). In addition, compound according to the present invention is suitable for being used as diuretic for the treatment of edema and electrolyte disturbances, especially in the situation of hypervolemic and normovolemic hyponatremia.

Furthermore, the compounds according to the invention can be used for the treatment and/or prevention of primary and secondary Raynaud's phenomenon, microcirculatory disorders, claudication, peripheral and autonomic neuropathies, diabetic microangiopathy, diabetic retinopathy, diabetic ulcerous limbs, gangrene, CREST syndrome, erythema, onychomycosis, rheumatic diseases and for promoting wound healing.

Furthermore, the compounds according to the invention are suitable for the treatment of urinary disorders and diseases of the urogenital system in men and women, for example, benign prostatic syndrome (BPS), benign prostatic hyperplasia (BPH), benign prostatic enlargement (BPE), bladder outlet obstruction (BOO), lower urinary tract syndrome (LUTS), neurogenic overactive bladder (OAB), interstitial cystitis (IC), urinary incontinence (UI), for example mixed, urge, stress and overflow urinary incontinence (MUI, UUI, SUI, OUI), pelvic pain, erectile dysfunction and female sexual dysfunction.

The compounds according to the present invention may also be used to treat and/or prevent 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, emphysema (e.g., emphysema caused by smoking), and cystic fibrosis (CF). In addition, the compounds of the present invention may be used to treat and/or prevent pulmonary arterial hypertension (PAH) and other forms of pulmonary hypertension (PH), including pulmonary hypertension associated with left ventricular disease, HIV infection, sickle cell anemia, thromboembolism (CTEPH), sarcoidosis, chronic obstructive pulmonary disease (COPD), or pulmonary fibrosis.

In addition, the compounds according to the invention can be used for the treatment and/or prevention of cirrhosis of the liver, ascites, diabetes and diabetic complications, such as neuropathy and nephropathy. In addition, the compounds according to the invention are suitable for the treatment and/or prevention of central nervous system disorders such as anxiety states and depression, glaucoma and cancer, in particular lung tumors, and for the management of circadian rhythm disorders, such as jet lag and shift work.

Furthermore, the compounds according to the invention can be used for the treatment and/or prevention of pain disorders, adrenal diseases such as pheochromocytoma and adrenal apoplexy, intestinal diseases such as Crohn's disease and diarrhea, menstrual disorders such as dysmenorrhea, or endometriosis, premature birth and for tocolysis.

Due to their activity and selectivity properties, 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 (types 1-5), hypervolemic and normovolemic hyponatremia, cirrhosis, ascites, edema and the syndrome of inappropriate ADH secretion (SIADH).

The above diseases have been well characterized in humans, but similar etiologies exist in other mammals, and these mammals can be treated using the compounds and methods of the present invention.

Therefore, the present invention also relates to the use of the compounds according to the invention for the treatment and/or prophylaxis of diseases, in particular the diseases mentioned above.

The present invention also relates to the use of the compounds according to the invention for the preparation of pharmaceutical compositions for the treatment and/or prevention of diseases, in particular the diseases mentioned above.

The present invention further relates to the use of the compounds according to the invention in a method for the treatment and/or prophylaxis of diseases, in particular the diseases mentioned above.

The present invention further relates to a method for treating and/or preventing diseases, in particular the diseases mentioned above, by using an effective amount of at least one compound according to the invention.

The compounds of the present invention can be administered in the form of a separate medicament, or can be administered in combination with one or more other therapeutic agents, as long as the administration of the combination does not result in undesirable and/or unacceptable side effects. Such combination therapy includes: administering a single pharmaceutical dosage form containing a compound of formula (I) as defined above and one or more other therapeutic agents, and administering the compound of formula (I) and each other preparation in its own separate pharmaceutical dosage form. For example, the compound of formula (I) and the therapeutic agent can be administered together to the patient in the form of a single (fixed) oral dosage composition such as a tablet or capsule, or each medicament can be administered in a separate dosage form.

When separate dosage forms are used, the compound of formula (I) and the one or more additional therapeutic agents may be administered at essentially the same time (ie, simultaneously) or at separately staggered times (ie, sequentially).

In particular, the compounds of the present invention can be used in fixed or separate combination with:

● Organic nitrates and NO donors, such as sodium nitroprusside, nitroglycerin, isosorbide mononitrate, isosorbide dinitrate, molsidomide, or SIN-1 and inhaled NO;

● compounds that inhibit the degradation of cyclic guanosine monophosphate (cGMP), for example inhibitors of phosphodiesterase (PDE) 1, 2 and/or 5, in particular PDE-5 inhibitors, such as sildenafil, vardenafil, tadalafil, udenafil, dasenafil, avanafil, milonafil or llodenafil;

● Positive inotropic drugs, such as cardiac glycosides (digoxin) and beta-adrenergic and dopaminergic agonists such as isoproterenol, epinephrine, norepinephrine, dopamine, or dobutamine;

● natriuretic peptides, such as atrial natriuretic peptide (ANP, anaritide), B-type natriuretic peptide or brain natriuretic peptide (BNP, nesiritide), C-type natriuretic peptide (CNP), or urodilidine;

● a calcium sensitizer, for example and preferably levosimendan;

• NO- and heme-independent soluble guanylate cyclase activators, such as, in particular, Cinaciguat and the compounds described in WO 01/19355, WO 01/19776, WO 01/19778, WO 01/19780, WO 02/070462 and WO 02/070510;

NO-independent, but heme-dependent guanylate cyclase stimulators, such as, in particular, riociguat, vericipirat and 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;

● Inhibitors of human neutrophil elastase (HNE), such as sivelestat or DX-890 (Reltran);

● compounds that inhibit signal transduction cascades, in particular tyrosine and/or serine/threonine kinase inhibitors, such as nintedanib, dasatinib, nilotinib, bosutinib, regorafenib, sorafenib, sunitinib, cediranib, axitinib, telatinib, imatinib, brivanib, pazopanib, vatalanib, gefitinib, erlotinib, lapatinib, canertinib, letutinib, pelitinib, semaxanib or tandutinib;

• compounds that influence cardiac energy metabolism, for example and preferably etomoxir, dichloroacetate, ranolazine or trimetazidine, or full or partial adenosine A1 receptor agonists;

• compounds that influence heart rate, for example and preferably ivabradine;

• a cardiac myosin activator, for example and preferably omecamtiv mecarbil (CK-1827452);

● an antithrombotic agent, for example and preferably selected from platelet aggregation inhibitors, anticoagulants and plasminogen substances;

● antihypertensive agents, for example and preferably selected from calcium antagonists, angiotensin AII antagonists, ACE inhibitors, vasopeptidase inhibitors, endothelin antagonists, renin inhibitors, alpha-blockers, beta-blockers, mineralocorticoid receptor antagonists and diuretics; and/or

● Agents that alter fat metabolism, for example and preferably selected from thyroid receptor agonists, cholesterol synthesis inhibitors (for example and preferably HMG-CoA-reductase or squalene synthesis inhibitors, ACAT inhibitors, CETP inhibitors, MTP inhibitors, PPAR-α, PPAR-γ and/or PPAR-δ agonists), cholesterol absorption inhibitors, lipase inhibitors, polymeric bile acid adsorbents, bile acid reabsorption inhibitors and lipoprotein (a) antagonists.

Antithrombotic agents are preferably understood to be compounds selected from the group consisting of platelet aggregation inhibitors, anticoagulants and plasminogen substances.

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.

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.

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.

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, fedexaban, razaxaban, fondaparinux, idroparux, 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.

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.

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.

Antihypertensive agents are preferably understood to mean compounds selected from the group consisting of calcium antagonists, angiotensin AII antagonists, ACE inhibitors, vasopeptidase inhibitors, endothelin antagonists, renin inhibitors, alpha-blockers, beta-blockers, mineralocorticoid receptor antagonists and diuretics.

In a preferred embodiment of the invention, the compounds according to the invention are administered in combination with a calcium antagonist, such as for example and preferably nifedipine, amlodipine, verapamil or diltiazem.

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.

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, adalol, landiolol, nebivolol, epanolol or bucindolol.

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.

In a preferred embodiment of the invention, the compounds according to the invention are administered in combination with a vasopeptidase inhibitor or neutral endopeptidase inhibitor (NEP), for example and preferably sacubitril, omapatrilat or AVE-7688.

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.

In a preferred embodiment of the invention, the compounds according to the invention are administered in combination with an ACE inhibitor, such as for example and preferably enalapril, captopril, lisinopril, ramipril, delapril, fosinopril, quinapril, perindopril or trandolapril.

In a preferred embodiment of the invention, the compounds according to the invention are administered in combination with an endothelin antagonist, such as for example and preferably bosentan, darusentan, ambrisentan, tezosentan or sitaxsentan.

In a preferred embodiment of the invention, the compounds according to the invention are administered in combination with a renin inhibitor, such as for example and preferably aliskiren, SPP-600 or SPP-800.

In a preferred embodiment of the invention, the compounds according to the invention are administered in combination with a mineralocorticoid receptor antagonist, such as for example and preferably finerenone, spironolactone, canrenone, potassium canrenoate or eplerenone.

In a preferred embodiment of the invention, the compounds according to the invention are administered in combination with a diuretic, for example and preferably furosemide, bumetanide, piretanide, torsemide, bendroflumethiazide, chlorothiazide, hydrochlorothiazide, xipamide, indapamide, hydroflumethiazide, methylclomethiazide, polythiazide, trichlorothiazide, chlorthalidone, metolazone, quinethazone, acetazolamide, dichlorbenzenesulfonamide, methazolamide, glycerol, isosorbide, mannitol, amiloride or triamterene.

Agents that alter fat metabolism are preferably understood to be compounds selected from the group consisting of CETP inhibitors, thyroid receptor agonists, cholesterol synthesis inhibitors (e.g. HMG-CoA reductase or squalene synthesis inhibitors, ACAT inhibitors, MTP inhibitors, PPAR-α, PPAR-γ and/or PPAR-δ agonists), cholesterol absorption inhibitors, polymeric bile acid adsorbents, bile acid reabsorption inhibitors, lipase inhibitors and lipoprotein (a) antagonists.

In a preferred embodiment of the invention, the compounds according to the invention are administered in combination with a CETP inhibitor, such as for example and preferably dalcetrapib, anacetrapib, BAY 60-5521 or CETP-vaccine (Avant).

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-thyroxine, 3,5,3'-triiodothyronine (T3), CGS 23425 or acitreol (CGS 26214).

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 group of the statins, for example and preferably lovastatin, simvastatin, pravastatin, fluvastatin, atorvastatin, rosuvastatin or pitavastatin.

In a preferred embodiment of the invention, the compounds according to the invention are administered in combination with a squalene synthesis inhibitor, such as for example and preferably BMS-188494 or TAK-475.

In a preferred embodiment of the invention, the compounds according to the invention are administered in combination with an ACAT inhibitor, such as for example and preferably avasimibe, melinamide, patiimibe, eflumimibe or SMP-797.

In a preferred embodiment of the invention, the compounds according to the invention are administered in combination with an MTP inhibitor, such as for example and preferably implitapide, R-103757, BMS-201038 or JTT-130.

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.

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.

In a preferred embodiment of the invention, the compounds according to the invention are administered in combination with a cholesterol absorption inhibitor, such as for example and preferably ezetimibe, tiqueside or pamaside.

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.

In a preferred embodiment of the invention, the compounds according to the invention are administered in combination with a polymeric bile acid adsorbent, for example and preferably cholestyramine, colestipol, colesolvam, cholesteat or colestilan.

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 an ASBT (= IBAT) inhibitor, such as, for example, AZD-7806, S-8921, AK-105, BARI-1741, SC-435 or SC-635.

In a preferred embodiment of the invention, the compounds according to the invention are administered in combination with a lipoprotein(a) antagonist, such as for example and preferably gemcabene calcium (CI-1027) or nicotinic acid.

In a particularly preferred embodiment, the compounds of the 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-blockers, mineralocorticoid receptor antagonists, organic nitrates, NO donors, soluble guanylate cyclase (SGC) activators, soluble guanylate cyclase stimulators and positive inotropic agents.

Therefore, in another embodiment, the present invention relates to pharmaceutical compositions comprising at least one compound according to the invention and one or more additional therapeutic agents for the treatment and/or prevention of diseases, in particular the aforementioned diseases.

Furthermore, the compounds of the present invention can be used as such, or in compositions, for research and diagnostics, or as analytical reference standards, etc., as is well known in the art.

When the compounds of the present invention are administered as medicines to humans or other mammals, they can be administered as such or in the form of pharmaceutical compositions containing, for example, 0.1% to 99.5% (more preferably 0.5% to 90%) of active ingredient and one or more pharmaceutically acceptable excipients.

Therefore, in a further aspect, the present invention relates to pharmaceutical compositions comprising at least one compound according to the invention and generally one or more inert, non-toxic, pharmaceutically suitable excipients, and to the use of such pharmaceutical compositions for the treatment and/or prevention of diseases, in particular the aforementioned diseases.

The compounds according to the present invention can act systemically and/or locally. For this purpose, they can be administered in a suitable manner, for example, orally, parenterally, through the lungs, through the nose, through the tongue, sublingually, through the buccal, rectal, dermal, transdermal, conjunctival, through the ear or local route, or as an implant or stent.

For these administration routes, the compounds according to the invention can be administered in suitable administration forms.

For oral administration, the following administration forms are suitable: the administration forms act according to the state of the art and deliver the compound according to the invention rapidly and/or in an improved manner and contain the compound according to the invention in crystalline form, amorphous form and/or dissolved form, such as tablets (uncoated or coated tablets, for example with an enteric coating or an insoluble or delayed dissolving coating which controls the release of the compound according to the invention), tablets which disintegrate rapidly in the mouth, or films/film tablets, films/lyophilizates, capsules (for example hard or soft gelatin capsules), sugar-coated tablets, granules, pills, powders, emulsions, suspensions, aerosols or solutions.

Parenteral administration can be carried out by bypassing the absorption step (intravenous, intraarterial, intracardial, intraspinal or intralumbar) or by including absorption (intramuscular, subcutaneous, intradermal, transdermal or intraperitoneal). Suitable parenteral administration forms include injection and infusion preparations in the form of solutions, suspensions, emulsions, lyophilizates or sterile powders.

Forms suitable for other routes of administration include, for example, inhalation forms (e.g., powder inhalers, nebulizers), nasal drops, solutions or sprays, tablets or capsules for translingual, sublingual or buccal administration (e.g., troches, lozenges), suppositories, otic and ophthalmic preparations (e.g., drops, ointments), vaginal capsules, aqueous suspensions (lotions, shake mixtures), lipophilic suspensions, ointments, creams, emulsions, pastes, foams, dusting powders, transdermal therapeutic systems (e.g., patches), implants and stents.

In a preferred embodiment, the pharmaceutical composition comprising a compound of formula (I) as described above is provided in a form suitable for oral administration. In another preferred embodiment, the pharmaceutical composition comprising a compound of formula (I) as described above is provided in a form suitable for intravenous administration.

The compounds of the invention can be converted into the administration forms in a manner known per se by mixing with inert, non-toxic, pharmaceutically suitable excipients, including in particular carriers (e.g., microcrystalline cellulose, lactose, mannitol), solvents (e.g., liquid polyethylene glycol), emulsifiers (e.g., sodium lauryl sulfate), surfactants (e.g., polyoxysorbitan oleate), dispersants (e.g., polyvinyl pyrrolidone), synthetic and natural polymers (e.g., albumin), stabilizers (e.g., antioxidants, e.g., ascorbic acid), colorants (e.g., inorganic pigments, e.g., iron oxide), and flavor and/or odor-masking agents.

The preferred dosage of the compounds of the present invention is the maximum amount that a patient can tolerate without experiencing serious side effects. Illustratively, the compounds of the present invention can be administered parenterally at a dosage of about 0.001 mg/kg body weight to about 10 mg/kg body weight, preferably about 0.01 mg/kg body weight to about 1 mg/kg body weight. When administered orally, exemplary dosage ranges are about 0.01 to 100 mg/kg body weight, preferably about 0.01 to 20 mg/kg body weight, and more preferably about 0.1 to 10 mg/kg body weight. Ranges intermediate to the above-recited values are also intended to be part of this invention.

However, actual dosage levels and administration schedules of the active ingredients in the pharmaceutical compositions of the present invention may be varied in order to obtain an amount of the active ingredient that is effective to achieve the desired therapeutic response for a particular patient, composition, and mode of administration without causing toxicity to the patient. Thus, it may be necessary to deviate from the stated amounts, where appropriate, depending, inter alia, on the age, sex, weight, diet, and general health of the patient; the bioavailability and pharmacokinetic properties of the particular compound and its mode and route of administration; the time or interval at which administration occurs; the dosing regimen selected; the individual patient's response to the active ingredient; the specific disease involved; the extent, involvement, or severity of the disease; the nature of concurrent therapy (i.e., the interaction of the compounds of the present invention with other concurrently administered therapeutic agents), and other relevant circumstances.

Thus, in some cases, less than the aforementioned minimum amount may be sufficient, while in other cases, the upper limit must be exceeded. Treatment can be started with a smaller dose that is less than the optimal dose of the compound. The dose can then be increased in small increments until the optimal effect in the circumstances is achieved. For convenience, the total daily dose can be divided and administered in separate portions spread over the day.

The following exemplary embodiments illustrate the present invention, but the present invention is not limited to these examples.

Unless otherwise indicated, percentages in the following tests and examples are percentages by weight and parts are parts by weight. Solvent ratios, dilution ratios and concentrations reported for liquid/liquid solutions are each based on volume.

A. Examples

Abbreviations and acronyms :

Ac acetyl

aq. aqueous (solution)

br. Broad peak ( ¹ H NMR signal)

cat. catalysis

conc. concentrated

d Doublet ( ¹ H NMR signal)

DCI Direct Chemical Ionization (MS)

d.e. diastereomeric excess

DMF N,N -dimethylformamide

DMSO dimethyl sulfoxide

EI Electron Impact Ionization (MS)

eq. equivalent

ESI Electrospray Ionization (MS)

Et ethyl

h hour

^1H NMR proton nuclear magnetic resonance spectroscopy

HPLC high-performance liquid chromatography

LC/MS liquid chromatography-mass spectrometry

m multiplet ( ^1H NMR signal)

Me methyl

min

MS

MTBE methyl tert-butyl ether

m/z mass-to-charge ratio (MS)

of th. Theoretical (chemical yield)

q quartet ( ¹ H NMR signal)

quant. quantitative (yield)

rac racemic

R _f TLC retention factor

RP Reversed Phase (HPLC)

rt room temperature

R _t retention time (HPLC)

s singlet ( ¹ H NMR signal)

sat. saturated (solution)

SFC Supercritical Fluid Chromatography

t triplet ( ¹ H NMR signal)

tBu tert-butyl

Uncle Tert

TFA trifluoroacetic acid

THF Tetrahydrofuran

TLC thin layer chromatography

UV ultraviolet rays.

LC/MS and HPLC methods :

Method 1 (LC/MS):

Instrument: Waters Acquity SQD UPLC system; Column: Waters Acquity UPLC HSS T3 1.8µ, 50 mm x 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 → 1.2 min 5% A → 2.0 min 5% A; Column oven: 50°C; Flow rate: 0.40 mL/min; UV detection: 208-400 nm.

Method 2 (LC/MS):

Instrument: Waters Acquity SQD UPLC system; Column: Waters Acquity UPLC HSS T3 1.8µ, 50 mm x 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 → 6.0 min 5% A → 7.5 min 5% A; Column oven: 50°C; Flow rate: 0.35 mL/min; UV detection: 210-400 nm.

Method 3 (LC/MS):

MS instrument: Agilent MS Quad 6150; HPLC instrument: Agilent 1290; column: Waters Acquity UPLC HSS T3 1.8 µ, 50 mm x 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 → 0.3 min 90% A →1.7 min 5% A → 3.0 min 5% A; column oven: 50°C; flow rate: 1.20 mL/min; UV detection: 205-305 nm.

Method 4 (preparative HPLC):

Column: Chromatorex C18 10 µm, 125 mm x 30 mm; Eluent A: water + 0.05% TFA, Eluent B: acetonitrile + 0.05% TFA; Gradient: 20% B → 45% B, 45% B isocratic, 45% B → 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

Under argon, potassium tert-butoxide (9.118 g, 81.26 mmol) was added portionwise to a solution of 5-(4-chlorophenyl)-4-[( 2S )-3,3,3-trifluoro-2-hydroxypropyl]-2,4-dihydro- 3H- 1,2,4-triazol-3-one (Example 5A in WO 2011/104322-A1; 20 g, 65.01 mmol) in THF (40 ml) at room temperature. Methyl bromoacetate (10.939 g, 71.51 mmol) was added to this solution, 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. This afforded 16.4 g (30.23 mmol) of the title compound (yield 46.5%, purity 70%).

The title compound can also be synthesized by 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-dihydro- 1H -1,2,4-triazol-1-yl}acetohydrazide

7.2 g (18.96 mmol) of 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 was dissolved in 60 ml of anhydrous ethanol. 2.088 g (41.71 mmol) of hydrazine hydrate were added to the solution, and the mixture was stirred under reflux for 5 h and then at room temperature overnight. The resulting mixture was partially concentrated under vacuum, then diluted with water and extracted with ethyl acetate. The combined organic phases were dried over sodium sulfate, filtered, and concentrated under vacuum. 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 target compound were obtained (yield 97.5%).

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

Under argon, sodium ethoxide (2.987 g, 42.14 mmol, 96% purity) was added portionwise 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) at room temperature. The reaction mixture was stirred at 100°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 afford 8.69 g (89% purity, 18.47 mmol) of the target compound, which was used without further purification (yield -88%).

Example 4A

5-(4-Chlorophenyl)-2-({5-[(1 RS )-1-hydroxyethyl]-1 H -1,2,4-triazol-3-yl}methyl)-4-[(2 S )-3,3,3-trifluoro-2-hydroxypropyl]-2,4-dihydro-3 H -1,2,4-triazol-3-one ( diastereoisomer mixture )

Under argon, sodium ethoxide (1.531 g, 21.59 mmol, 96% purity) was added portionwise 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-hydroxypropionamidine hydrochloride (1.480 g, 11.88 mmol) in DMF (110 ml) at room temperature. The reaction mixture was stirred at 120°C for 4.5 hours. 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 target compound (diastereoisomer mixture) which was used without further purification.

Example 5A

5-(4-Chlorophenyl)-2-({5-[(1 S )-1-hydroxyethyl]-1 H -1,2,4-triazol-3-yl}methyl)-4-[(2 S )-3,3,3-trifluoro-2-hydroxypropyl]-2,4-dihydro-3 H -1,2,4-triazol-3-one

The title compound was synthesized analogously to Example 4A starting from ( 2S )-2-hydroxypropionamidine hydrochloride (1.1 eq.) and using only 1.1 eq. of sodium ethoxide as base (reaction temperature: 100°C).

Commercially available ( 2S )-2-hydroxypropionamidine hydrochloride can also be synthesized from ( 2S )-2-hydroxypropionamide [1-(-)-lactamide] by the procedures described in WO 00/59510-A1 (Preparation 11, Step A) and WO 2013/138860-A1 (Intermediate of Precursor 82, p. 101-102).

Example 6A

5-(4-Chlorophenyl)-2-({5-[(1 R )-1-hydroxyethyl]-1 H -1,2,4-triazol-3-yl}methyl)-4-[(2 S )-3,3,3-trifluoro-2-hydroxypropyl]-2,4-dihydro-3 H -1,2,4-triazol-3-one

The title compound was synthesized analogously to Example 4A starting from ( 2R )-2-hydroxypropionamidine hydrochloride (1.1 eq.) and using only 1.1 eq. of sodium ethoxide as base (reaction temperature: 100°C).

Commercially available ( 2R )-2-hydroxypropionamidine hydrochloride can also be synthesized from ( 2R )-2-hydroxypropionamide [R-(+)-lactamide] by the procedures described in WO 00/59510-A1 (Preparation 11, Step A) and WO 2013/138860-A1 (Intermediate of Precursor 82, p. 101-102).

Preparation Example :

The example compounds with 1-hydroxyethyl substituents [R ¹ = CH ₃ in formula (I)], hereinafter referred to as "diastereomer 1" and "diastereomer 2", respectively, represent a pair of separated diastereomers, the absolute configuration (1 R or 1 S ) of which about the 1-hydroxyethyl moiety has not been determined.

The diastereomeric excess (d.e.) values were determined in a conventional manner by HPLC peak area analysis according to the following formula:

.

Example 1

5-(4-chlorophenyl)-2-{[1-(3-chlorophenyl)-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

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) was 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, then, due to incomplete conversion, additional boronic acid (74.7 mg, 0.48 mmol, 0.5 eq.) was added. After stirring for an additional 2 days, the reaction mixture was 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] to afford the title compound (113 mg, 0.21 mmol) (yield 22.4%).

Example 2

5-(4-chlorophenyl)-2-{[1-(3-fluorophenyl)-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

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) was 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, then, due to incomplete conversion, additional boronic acid (16.72 mg, 0.12 mmol, 0.5 eq.) was added. After stirring for an additional 2 days, the reaction mixture was 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] to afford the title compound (25 mg, 0.05 mmol) (yield 20.2%).

Example 3

5-(4-chlorophenyl)-2-{[5-(hydroxymethyl)-1-(2-methoxyphenyl) -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

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) was 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, then, due to incomplete conversion, additional boronic acid (54.4 mg, 0.36 mmol, 0.5 eq.) was added. After stirring for an additional 2 days, the reaction mixture was 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] to afford the title compound (73 mg, 0.14 mmol) (yield 22.4%, purity 98%).

Example 4

5-(4-chlorophenyl)-2-{[1-(2-chlorophenyl)-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

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) was 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, then, due to incomplete conversion, additional boronic acid (500 mg, 3.20 mmol) was added. The reaction mixture was stirred at room temperature for an additional 9 days. During this time, 3 more portions of boronic acid (total 1.5 g, 9.6 mmol) were added. The reaction mixture was 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] to obtain the target compound (1.44 g, 2.72 mmol) (yield 45.7%).

Example 5

5-(4-Chlorophenyl)-2-{[5-(hydroxymethyl)-1-(2-methylphenyl) -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

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) was 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, then, due to incomplete conversion, additional boronic acid (64.9 mg, 0.48 mmol, 0.5 eq.) was added. After stirring for an additional 2 days, the reaction mixture was 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] to afford the title compound (58 mg, 0.11 mmol) (yield 11.9%).

Example 6

2-{[1-(3-chloro-5-fluorophenyl)-5-(hydroxymethyl) -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

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) was 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, then, due to incomplete conversion, additional boronic acid (83.2 mg, 0.48 mmol, 0.5 eq.) was added. After stirring for an additional 2 days, the reaction mixture was 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] to afford the title compound (91 mg, 0.17 mmol) (yield 17.4%).

Example 7

5-(4-chlorophenyl)-2-{[1-(3,5-difluorophenyl)-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

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. This gave 61 mg (0.11 mmol) of the target compound (yield 13.4%).

Example 8

5-(4-chlorophenyl)-2-({5-(hydroxymethyl)-1-[2-(trifluoromethyl)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

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. This gave 104 mg (0.18 mmol) of the target compound (yield 15.5%).

Example 9

2-{[1-(2-chloro-5-fluorophenyl)-5-(hydroxymethyl) -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

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. This gave 8.7 mg (0.02 mmol) of the target compound (yield 1.3%, purity 95%).

Example 10

5-(4-Chlorophenyl)-2-({5-[(1 RS )-1-hydroxyethyl]-1-(2-methoxyphenyl)-1 H -1,2,4-triazol-3-yl}methyl)-4-[(2 S )-3,3,3-trifluoro-2-hydroxypropyl]-2,4-dihydro-3 H -1,2,4-triazol-3-one ( diastereoisomer mixture )

To a solution of 5-(4-chlorophenyl)-2-({5-[(1 RS )-1-hydroxyethyl]-1 H -1,2,4-triazol-3-yl}methyl)-4-[(2 S )-3,3,3-trifluoro-2-hydroxypropyl]-2,4-dihydro-3 H -1,2,4-triazol-3-one (500 mg, 1.16 mmol) in pyridine (15 ml) was 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, then, due to incomplete conversion, additional boronic acid (87 mg, 0.58 mmol, 0.5 eq.) was added. After stirring for an additional 2 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] to afford the title compound (132 mg, 0.22 mmol) as a mixture of diastereomers (yield 19.1%, purity 90%).

The two diastereomers were separated by preparative chiral HPLC [sample preparation: dissolve 128 mg in 10 ml of methanol; injection volume: 0.3 ml; column: Daicel ^Chiralcel® OX-H 5 µm, 250 x 20 mm; eluent: isohexane/methanol 70:30; flow rate: 80 ml/min; temperature: 40°C; UV detection: 210 nm]. After separation, 43.6 mg of the first-eluting diastereomer 1 (Example 11) and 45.1 mg of the second-eluting diastereomer 2 (Example 12) were isolated.

Example 11

5-(4-Chlorophenyl)-2-{[5-(1-hydroxyethyl)-1-(2-methoxyphenyl) -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 )

Analytical chiral HPLC (SFC): R _t = 3.08 min, de = 100% [column: Daicel Chiralcel ^® OX-3 250 x 4 mm; eluent: carbon dioxide/methanol (5% → 60%); flow rate: 3 ml/min; UV detection: 220 nm].

Example 12

5-(4-Chlorophenyl)-2-{[5-(1-hydroxyethyl)-1-(2-methoxyphenyl) -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 )

Analytical chiral HPLC (SFC): R _t = 3.38 min, de = 91.1% [column: Daicel Chiralcel ^® OX-3 250 x 4 mm; eluent: carbon dioxide/methanol (5% → 60%); flow rate: 3 ml/min; UV detection: 220 nm].

Example 13

2-({1-(3-chloro-5-fluorophenyl)-5-[(1 RS )-1-hydroxyethyl]-1 H -1,2,4-triazol-3-yl}methyl)-5-(4-chlorophenyl)-4-[(2 S )-3,3,3-trifluoro-2-hydroxypropyl]-2,4-dihydro-3 H -1,2,4-triazol-3-one ( diastereoisomer mixture)

To a solution of 5-(4-chlorophenyl)-2-({5-[(1 RS )-1-hydroxyethyl]-1 H -1,2,4-triazol-3-yl}methyl)-4-[(2 S )-3,3,3-trifluoro-2-hydroxypropyl]-2,4-dihydro-3 H -1,2,4-triazol-3-one (430 mg, 0.99 mmol) in pyridine (12.5 ml) was 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, then additional boronic acid (86.7 mg, 0.497 mmol, 0.5 eq.) was added due to incomplete conversion. After stirring for another 2 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] to obtain the target compound (148 mg, 0.26 mmol) as a mixture of diastereomers (yield 26.5%).

The two diastereomers were separated by preparative chiral HPLC (SFC) [sample preparation: dissolving 143 mg in 15 ml of methanol; injection volume: 0.5 ml; column: Daicel ^Chiralcel® OX-H 5 µm, 250 x 20 mm; eluent: carbon dioxide/methanol 80:20; flow rate: 80 ml/min; temperature: 40°C; UV detection: 210 nm]. After separation, 70 mg of the first-eluting diastereomer 1 (Example 14) and 60 mg of the second-eluting diastereomer 2 (Example 15) were isolated.

Example 14

2-{[1-(3-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-dihydro- 3H -1,2,4-triazol-3-one ( Diastereomer 1 )

Analytical chiral HPLC (SFC): R _t = 4.45 min, de = 100% [column: Daicel Chiralcel ^® OX-3 250 x 4 mm; eluent: carbon dioxide/methanol (5% → 60%); flow rate: 3 ml/min; UV detection: 220 nm].

Example 15

2-{[1-(3-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-dihydro- 3H -1,2,4-triazol-3-one ( diastereomer 2 )

Analytical chiral HPLC (SFC): R _t = 4.80 min, de = 100% [column: Daicel Chiralcel ^® OX-3 250 x 4 mm; eluent: carbon dioxide/methanol (5% → 60%); flow rate: 3 ml/min; UV detection: 220 nm].

Example 16

5-(4-Chlorophenyl)-2-({5-[(1 RS )-1-hydroxyethyl]-1-phenyl-1 H -1,2,4-triazol-3-yl}methyl)-4-[(2 S )-3,3,3-trifluoro-2-hydroxypropyl]-2,4-dihydro-3 H -1,2,4-triazol-3-one ( diastereoisomer mixture )

To a solution of 5-(4-chlorophenyl)-2-({5-[(1 RS )-1-hydroxyethyl]-1 H -1,2,4-triazol-3-yl}methyl)-4-[(2 S )-3,3,3-trifluoro-2-hydroxypropyl]-2,4-dihydro-3 H -1,2,4-triazol-3-one (250 mg, 0.462 mmol, 80% purity) in pyridine (6 ml) was 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° C. for 2 h and then stirred at room temperature for 4 days. Then, due to incomplete conversion, additional boronic acid (28.2 mg, 0.23 mmol, 0.5 eq.) was added. After stirring at 60°C for another 4 h and then at room temperature for another 2 days, the reaction mixture was concentrated in vacuo, 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] to obtain the target compound (42.5 mg, 0.08 mmol) as a mixture of diastereomers (yield 18.1%).

The two diastereomers were separated by preparative chiral HPLC [sample preparation: 38.9 mg dissolved in 1 ml of ethanol/isohexane (1:1); injection volume: 1 ml; column: Daicel ^Chiralcel® OX-H 5 µm, 250 x 20 mm; eluent: isohexane/ethanol 75:25; flow rate: 15 ml/min; temperature: 30°C; UV detection: 220 nm]. After separation, 13 mg of the first-eluting diastereomer 1 (Example 17) and 14 mg of the second-eluting diastereomer 2 (Example 18) were isolated.

Example 17

5-(4-Chlorophenyl)-2-{[5-(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 ( Diastereomer 1 )

Analytical chiral HPLC: R _t = 8.18 min, de = 100% [column: LUX Cellulose-4, 5 µm, 250 x 4.6 mm; eluent: isohexane/ethanol 70:30; flow rate: 1 ml/min; temperature: 40°C; UV detection: 220 nm].

Example 18

5-(4-Chlorophenyl)-2-{[5-(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 ( Diastereomer 2 )

Analytical chiral HPLC: R _t = 11.40 min, de = 100% [column: LUX Cellulose-4, 5 µm, 250 x 4.6 mm; eluent: isohexane/ethanol 70:30; flow rate: 1 ml/min; temperature: 40°C; UV detection: 220 nm].

Example 19

5-(4-Chlorophenyl)-2-({1-[3-(difluoromethyl)phenyl]-5-[(1 RS )-1-hydroxyethyl]-1 H -1,2,4-triazol-3-yl}methyl)-4-[(2 S )-3,3,3-trifluoro-2-hydroxypropyl]-2,4-dihydro-3 H -1,2,4-triazol-3-one ( diastereoisomer mixture )

To a solution of 5-(4-chlorophenyl)-2-({5-[(1 RS )-1-hydroxyethyl]-1 H -1,2,4-triazol-3-yl}methyl)-4-[(2 S )-3,3,3-trifluoro-2-hydroxypropyl]-2,4-dihydro-3 H -1,2,4-triazol-3-one (400 mg, 0.74 mmol, 80% purity) in pyridine (9.6 ml) was 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° 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] to afford the title compound (47 mg, 0.08 mmol) as a mixture of diastereomers (yield 11.3%).

The two diastereomers were separated by preparative chiral HPLC [sample preparation: 45 mg dissolved in 1 ml of ethanol/isohexane (1:1); injection volume: 1 ml; column: Daicel ^Chiralcel® OX-H 5 µm, 250 x 20 mm; eluent: isohexane/ethanol 75:25; flow rate: 15 ml/min; temperature: 30°C; UV detection: 220 nm]. After separation, 20 mg of the first-eluting diastereomer 1 (Example 20) and 20 mg of the second-eluting diastereomer 2 (Example 21) 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-dihydro- 3H -1,2,4-triazol-3-one ( Diastereomer 1 )

Analytical chiral HPLC: R _t = 6.72 min, de = 99% [column: LUX Cellulose-4, 5 µm, 250 x 4.6 mm; eluent: isohexane/ethanol 70:30; flow rate: 1 ml/min; temperature: 40°C; UV detection: 220 nm].

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-dihydro- 3H -1,2,4-triazol-3-one ( Diastereomer 2 )

Analytical chiral HPLC: R _t = 9.36 min, de = 100% [column: LUX Cellulose-4, 5 µm, 250 x 4.6 mm; eluent: isohexane/ethanol 70:30; flow rate: 1 ml/min; temperature: 40°C; UV detection: 220 nm].

Example 22

5-(4-Chlorophenyl)-2-({5-[(1 RS )-1-hydroxyethyl]-1-[3-(trifluoromethyl)phenyl]-1 H -1,2,4-triazol-3-yl}methyl)-4-[(2 S )-3,3,3-trifluoro-2-hydroxypropyl]-2,4-dihydro-3 H -1,2,4-triazol-3-one ( diastereoisomer mixture )

To a solution of 5-(4-chlorophenyl)-2-({5-[(1 RS )-1-hydroxyethyl]-1 H -1,2,4-triazol-3-yl}methyl)-4-[(2 S )-3,3,3-trifluoro-2-hydroxypropyl]-2,4-dihydro-3 H -1,2,4-triazol-3-one (400 mg, 0.74 mmol, 80% purity) in pyridine (9.6 ml) was 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° C. for 2 h and then stirred at room temperature for 5 days. Then, due to incomplete conversion, additional boronic acid (70.2 mg, 0.37 mmol, 0.5 eq.) was added. After stirring at 60°C for another 4 h and then at room temperature overnight, the reaction mixture was concentrated in vacuo, 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] to obtain the target compound (58.2 mg, 0.10 mmol) as a mixture of diastereomers (yield 13.6%).

The two diastereomers were separated by preparative chiral HPLC [sample preparation: 58 mg dissolved in 2 ml of ethanol/isohexane (1:1); injection volume: 1 ml; column: Daicel ^Chiralcel® OX-H 5 µm, 250 x 20 mm; eluent: isohexane/ethanol 75:25; flow rate: 15 ml/min; temperature: 30°C; UV detection: 220 nm]. After separation, 19.5 mg of the first-eluting diastereomer 1 (Example 23) and 19.2 mg of the second-eluting diastereomer 2 (Example 24) were isolated.

Example 23

5-(4-Chlorophenyl)-2-({5-(1-hydroxyethyl)-1-[3-(trifluoromethyl)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 ( Diastereomer 1 )

Analytical chiral HPLC: R _t = 4.94 min, de = 100% [column: LUX Cellulose-4, 5 µm, 250 x 4.6 mm; eluent: isohexane/ethanol 70:30; flow rate: 1 ml/min; temperature: 40°C; UV detection: 220 nm].

Example 24

5-(4-Chlorophenyl)-2-({5-(1-hydroxyethyl)-1-[3-(trifluoromethyl)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 ( Diastereomer 2 )

Analytical chiral HPLC: R _t = 6.13 min, de = 98.6% [column: LUX Cellulose-4, 5 µm, 250 x 4.6 mm; eluent: isohexane/ethanol 70:30; flow rate: 1 ml/min; temperature: 40°C; UV detection: 220 nm].

Example 25

5-(4-Chlorophenyl)-2-({1-(3,5-difluorophenyl)-5-[(1 RS )-1-hydroxyethyl]-1 H -1,2,4-triazol-3-yl}methyl)-4-[(2 S )-3,3,3-trifluoro-2-hydroxypropyl]-2,4-dihydro-3 H -1,2,4-triazol-3-one ( diastereoisomer mixture )

To a solution of 5-(4-chlorophenyl)-2-({5-[(1 RS )-1-hydroxyethyl]-1 H -1,2,4-triazol-3-yl}methyl)-4-[(2 S )-3,3,3-trifluoro-2-hydroxypropyl]-2,4-dihydro-3 H -1,2,4-triazol-3-one (400 mg, 0.92 mmol) in pyridine (12 ml) was 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° C. for 2 h and then stirred at room temperature for 4 days. Then, due to incomplete conversion, additional boronic acid (72.98 mg, 0.46 mmol, 0.5 eq.) was added. After stirring at 60°C for another 2 h and then at room temperature for another 3 days, the reaction mixture was concentrated in vacuo, 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] to obtain the target compound (114.3 mg, 0.21 mmol) as a mixture of diastereomers (yield 22.7%).

The two diastereomers were separated by preparative chiral HPLC [sample preparation: 110 mg dissolved in 7 ml of ethanol/isohexane (1:1); injection volume: 0.6 ml; column: Daicel ^Chiralcel® OX-H 5 µm, 250 x 20 mm; eluent: isohexane/ethanol 70:30; flow rate: 20 ml/min; temperature: 40°C; UV detection: 220 nm]. After separation, 42 mg of the first-eluting diastereomer 1 (Example 26) and 44 mg of the second-eluting diastereomer 2 (Example 27) 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 )

Analytical chiral HPLC: R _t = 1.09 min, de = 100% [column: Daicel Chiralpack OX-3 3µm, 50 x 4.6 mm; eluent: isohexane/ethanol 70:30; flow rate: 1 ml/min; UV detection: 220 nm].

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 )

Analytical chiral HPLC: R _t = 1.28 min, de = 99% [column: Daicel Chiralpack OX-3 3 µm, 50 x 4.6 mm; eluent: isohexane/ethanol 70:30; flow rate: 1 ml/min; UV detection: 220 nm].

Example 28

5-(4-Chlorophenyl)-2-({5-[(1 RS )-1-hydroxyethyl]-1-(3-methylphenyl)-1 H -1,2,4-triazol-3-yl}methyl)-4-[(2 S )-3,3,3-trifluoro-2-hydroxypropyl]-2,4-dihydro-3 H -1,2,4-triazol-3-one ( diastereoisomer mixture )

To a solution of 5-(4-chlorophenyl)-2-({5-[(1 RS )-1-hydroxyethyl]-1 H -1,2,4-triazol-3-yl}methyl)-4-[(2 S )-3,3,3-trifluoro-2-hydroxypropyl]-2,4-dihydro-3 H -1,2,4-triazol-3-one (400 mg, 0.92 mmol) in pyridine (12 ml) was 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° C. for 2 h and then stirred at room temperature for 3 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] to give the target compound (59.6 mg, 0.11 mmol) as a mixture of diastereomers (yield 12.3%).

The two diastereomers were separated by preparative chiral HPLC [sample preparation: 56 mg dissolved in 2 ml of ethanol/isohexane (1:1); injection volume: 0.5 ml; column: Daicel ^Chiralcel® OX-H 5 µm, 250 x 20 mm; eluent: isohexane/ethanol 70:30; flow rate: 15 ml/min; temperature: 25°C; UV detection: 220 nm]. After separation, 22 mg of the first-eluting diastereomer 1 (Example 29) and 24 mg of the second-eluting diastereomer 2 (Example 30) were isolated.

Example 29

5-(4-Chlorophenyl)-2-{[5-(1-hydroxyethyl)-1-(3-methylphenyl) -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 )

Analytical chiral HPLC: R _t = 7.97 min, de = 100% [column: LUX Cellulose-4, 5 µm, 250 x 4.6 mm; eluent: isohexane/ethanol 70:30; flow rate: 1 ml/min; temperature: 40°C; UV detection: 220 nm].

Example 30

5-(4-Chlorophenyl)-2-{[5-(1-hydroxyethyl)-1-(3-methylphenyl) -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 )

Analytical chiral HPLC: R _t = 11.44 min, de = 99.1% [column: LUX Cellulose-4, 5 µm, 250 x 4.6 mm; eluent: isohexane/ethanol 70:30; flow rate: 1 ml/min; temperature: 40°C; UV detection: 220 nm].

Example 31

5-(4-Chlorophenyl)-2-({1-(2-ethylphenyl)-5-[(1 RS )-1-hydroxyethyl]-1 H -1,2,4-triazol-3-yl}methyl)-4-[(2 S )-3,3,3-trifluoro-2-hydroxypropyl]-2,4-dihydro-3 H -1,2,4-triazol-3-one ( diastereoisomer mixture )

To a solution of 5-(4-chlorophenyl)-2-({5-[(1 RS )-1-hydroxyethyl]-1 H -1,2,4-triazol-3-yl}methyl)-4-[(2 S )-3,3,3-trifluoro-2-hydroxypropyl]-2,4-dihydro-3 H -1,2,4-triazol-3-one (600 mg, 1.39 mmol) in pyridine (18 ml) was 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° C. for 2 h and then stirred at room temperature for 3 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] to give the target compound (69.4 mg, 0.13 mmol) as a mixture of diastereomers (yield 9.1%).

The two diastereomers were separated by preparative chiral HPLC [sample preparation: 65 mg dissolved in 4 ml of ethanol/isohexane (1:1); injection volume: 0.5 ml; column: Daicel ^Chiralcel® OX-H 5 µm, 250 x 20 mm; eluent: isohexane/ethanol 50:50; flow rate: 20 ml/min; temperature: 40°C; UV detection: 220 nm]. After separation, 25 mg of the first-eluting diastereomer 1 (Example 32) and 25 mg of the second-eluting diastereomer 2 (Example 33) were isolated.

Example 32

5-(4-Chlorophenyl)-2-{[1-(2-ethylphenyl)-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 )

Analytical chiral HPLC: R _t = 0.96 min, de = 100% [column: Daicel Chiralpack OX-3 3µm, 50 x 4.6 mm; eluent: isohexane/ethanol 50:50; flow rate: 1 ml/min; UV detection: 220 nm].

Example 33

5-(4-Chlorophenyl)-2-{[1-(2-ethylphenyl)-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 )

Analytical chiral HPLC: R _t = 1.09 min, de = 100% [column: Daicel Chiralpack OX-3 3µm, 50 x 4.6 mm; eluent: isohexane/ethanol 50:50; flow rate: 1 ml/min; UV detection: 220 nm].

Example 34

2-({1-(2-chloro-4-fluorophenyl)-5-[(1 RS )-1-hydroxyethyl]-1 H -1,2,4-triazol-3-yl}methyl)-5-(4-chlorophenyl)-4-[(2 S )-3,3,3-trifluoro-2-hydroxypropyl]-2,4-dihydro-3 H -1,2,4-triazol-3-one ( diastereoisomer mixture)

To a solution of 5-(4-chlorophenyl)-2-({5-[(1 RS )-1-hydroxyethyl]-1 H -1,2,4-triazol-3-yl}methyl)-4-[(2 S )-3,3,3-trifluoro-2-hydroxypropyl]-2,4-dihydro-3 H -1,2,4-triazol-3-one (600 mg, 1.39 mmol) in pyridine (18 ml) was 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° C. for 2 h and then stirred at room temperature for 5 days. Then, due to incomplete conversion, additional boronic acid (242 mg, 1.39 mmol) was added. The reaction mixture was stirred at room temperature for an additional 4 days. During this time, two more portions of boronic acid were added (483 mg, 2.77 mmol in total). The reaction mixture was then concentrated in vacuo, 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 target compound (107 mg, 0.19 mmol) was obtained as a mixture of diastereomers (yield 13.7%).

The two diastereomers were separated by preparative chiral HPLC [sample preparation: 104 mg dissolved in 5 ml of ethanol/isohexane (1:1); injection volume: 0.5 ml; column: Daicel ^Chiralcel® OX-H 5 µm, 250 x 20 mm; eluent: isohexane/ethanol 70:30; flow rate: 20 ml/min; temperature: 40°C; UV detection: 220 nm]. After separation, 56 mg of the first-eluting diastereomer 1 (Example 35) and 29 mg of the second-eluting diastereomer 2 (Example 36) were isolated.

Example 35

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-dihydro- 3H -1,2,4-triazol-3-one ( Diastereomer 1 )

Analytical chiral HPLC: R _t = 1.36 min, de = 100% [column: Daicel Chiralpack OX-3 3µm, 50 x 4.6 mm; eluent: isohexane/ethanol 70:30; flow rate: 1 ml/min; UV detection: 220 nm].

Example 36

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-dihydro- 3H -1,2,4-triazol-3-one ( diastereomer 2 )

Analytical chiral HPLC: R _t = 1.72 min, de = 100% [column: Daicel Chiralpack OX-3 3µm, 50 x 4.6 mm; eluent: isohexane/ethanol 70:30; flow rate: 1 ml/min; UV detection: 220 nm].

Example 37

5-(4-Chlorophenyl)-2-({1-(5-fluoro-2-methoxyphenyl)-5-[(1 RS )-1-hydroxyethyl]-1 H -1,2,4-triazol-3-yl}methyl)-4-[(2 S )-3,3,3-trifluoro-2-hydroxypropyl]-2,4-dihydro-3 H -1,2,4-triazol-3-one ( diastereoisomer mixture )

To a solution of 5-(4-chlorophenyl)-2-({5-[(1 RS )-1-hydroxyethyl]-1 H -1,2,4-triazol-3-yl}methyl)-4-[(2 S )-3,3,3-trifluoro-2-hydroxypropyl]-2,4-dihydro-3 H -1,2,4-triazol-3-one (600 mg, 1.39 mmol) in pyridine (18 ml) was 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° C. for 2 h and then stirred at room temperature for 3 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] to give the target compound (62.2 mg, 0.11 mmol) as a mixture of diastereomers (yield 8.1%).

The two diastereomers were separated by preparative chiral HPLC [sample preparation: 55.4 mg dissolved in 6 ml of ethanol/isohexane (1:1); injection volume: 2 ml; column: Daicel ^Chiralcel® OX-H 5 µm, 250 x 20 mm; eluent: isohexane/ethanol 80:20; flow rate: 20 ml/min; temperature: 40°C; UV detection: 220 nm]. After separation, 23 mg of the first-eluting diastereomer 1 (Example 38) and 21 mg of the second-eluting diastereomer 2 (Example 39) 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-dihydro- 3H -1,2,4-triazol-3-one ( Diastereomer 1 )

Analytical chiral HPLC: R _t = 2.13 min, de = 100% [column: Daicel Chiralpack OX-3 3µm, 50 x 4.6 mm; eluent: isohexane/ethanol 80:20; flow rate: 1 ml/min; temperature: 30°C; UV detection: 220 nm].

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-dihydro- 3H -1,2,4-triazol-3-one ( Diastereomer 2 )

Analytical chiral HPLC: R _t = 2.75 min, de = 100% [column: Daicel Chiralpack OX-3 3µm, 50 x 4.6 mm; eluent: isohexane/ethanol 80:20; flow rate: 1 ml/min; temperature: 30°C; UV detection: 220 nm].

Example 40

5-(4-Chlorophenyl)-2-({1-(2-fluorophenyl)-5-[(1 RS )-1-hydroxyethyl]-1 H -1,2,4-triazol-3-yl}methyl)-4-[(2 S )-3,3,3-trifluoro-2-hydroxypropyl]-2,4-dihydro-3 H -1,2,4-triazol-3-one ( diastereoisomer mixture )

To a solution of 5-(4-chlorophenyl)-2-({5-[(1 RS )-1-hydroxyethyl]-1 H -1,2,4-triazol-3-yl}methyl)-4-[(2 S )-3,3,3-trifluoro-2-hydroxypropyl]-2,4-dihydro-3 H -1,2,4-triazol-3-one (600 mg, 1.39 mmol) in pyridine (18 ml) was 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° C. for 2 h and then stirred at room temperature for 3 days. During this time, two additional portions of boronic acid were added (total 387.96 mg, 2.77 mmol). The resulting reaction mixture was then concentrated in vacuo, 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] to afford the title compound (30.1 mg, 0.06 mmol) as a mixture of diastereomers (yield 4.1%).

The two diastereomers were separated by preparative chiral HPLC [sample preparation: 26 mg dissolved in 4 ml of ethanol/isohexane (1:1); injection volume: 2 ml; column: Daicel ^Chiralcel® OX-H 5 µm, 250 x 20 mm; eluent: isohexane/ethanol 80:20; flow rate: 25 ml/min; temperature: 40°C; UV detection: 220 nm]. After separation, 11 mg of the first-eluting diastereomer 1 (Example 41) and 9 mg of the second-eluting diastereomer 2 (Example 42) were isolated.

Example 41

5-(4-Chlorophenyl)-2-{[1-(2-fluorophenyl)-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 )

Analytical chiral HPLC: R _t = 2.32 min, de = 100% [column: Daicel Chiralpack OX-3 3µm, 50 x 4.6 mm; eluent: isohexane/ethanol 80:20; flow rate: 1 ml/min; temperature: 30°C; UV detection: 220 nm].

Example 42

5-(4-Chlorophenyl)-2-{[1-(2-fluorophenyl)-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 )

Analytical chiral HPLC: R _t = 3.23 min, de = 100% [column: Daicel Chiralpack OX-3 3µm, 50 x 4.6 mm; eluent: isohexane/ethanol 80:20; flow rate: 1 ml/min; temperature: 30°C; UV detection: 220 nm].

Example 43

2-({1-(3-chloro-4-fluorophenyl)-5-[(1 RS )-1-hydroxyethyl]-1 H -1,2,4-triazol-3-yl}methyl)-5-(4-chlorophenyl)-4-[(2 S )-3,3,3-trifluoro-2-hydroxypropyl]-2,4-dihydro-3 H -1,2,4-triazol-3-one ( diastereoisomer mixture)

To a solution of 5-(4-chlorophenyl)-2-({5-[(1 RS )-1-hydroxyethyl]-1 H -1,2,4-triazol-3-yl}methyl)-4-[(2 S )-3,3,3-trifluoro-2-hydroxypropyl]-2,4-dihydro-3 H -1,2,4-triazol-3-one (400 mg, 0.92 mmol) in pyridine (12 ml) was 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° 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] to give the target compound (99.1 mg, 0.18 mmol) as a mixture of diastereomers (yield 19.1%).

The two diastereomers were separated by preparative chiral HPLC [sample preparation: 97.1 mg dissolved in 3 ml of ethanol; injection volume: 0.3 ml; column: Daicel ^Chiralcel® OX-H 5 µm, 250 x 20 mm; eluent: isohexane/ethanol 80:20; flow rate: 15 ml/min; temperature: 25°C; UV detection: 220 nm]. After separation, 40 mg of the first-eluting diastereomer 1 (Example 44) and 42 mg of the second-eluting diastereomer 2 (Example 45) were isolated.

Example 44

2-{[1-(3-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-dihydro- 3H -1,2,4-triazol-3-one ( Diastereomer 1 )

Preparative chiral HPLC: R _t = 9.97 min, de = 100% [column: Daicel Chiralcel ^® OX-H 5µm, 250 x 20 mm; eluent: isohexane/ethanol 80:20; flow rate: 15 ml/min; temperature: 25°C; UV detection: 220 nm].

Example 45

2-{[1-(3-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-dihydro- 3H -1,2,4-triazol-3-one ( diastereomer 2 )

Preparative chiral HPLC: R _t = 11.35 min, de = 100% [column: Daicel Chiralcel ^® OX-H 5µm, 250 x 20 mm; eluent: isohexane/ethanol 80:20; flow rate: 15 ml/min; temperature: 25°C; UV detection: 220 nm].

Example 46

5-(4-Chlorophenyl)-2-({1-(3,5-dichlorophenyl)-5-[(1 RS )-1-hydroxyethyl]-1 H -1,2,4-triazol-3-yl}methyl)-4-[(2 S )-3,3,3-trifluoro-2-hydroxypropyl]-2,4-dihydro-3 H -1,2,4-triazol-3-one ( diastereoisomer mixture )

To a solution of 5-(4-chlorophenyl)-2-({5-[(1 RS )-1-hydroxyethyl]-1 H -1,2,4-triazol-3-yl}methyl)-4-[(2 S )-3,3,3-trifluoro-2-hydroxypropyl]-2,4-dihydro-3 H -1,2,4-triazol-3-one (400 mg, 0.92 mmol) in pyridine (12 ml) was 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° 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] to give the target compound (105.5 mg, 0.18 mmol) as a mixture of diastereomers (yield 19.8%).

The two diastereomers were separated by preparative chiral HPLC [sample preparation: 103.5 mg dissolved in 14 ml of ethanol/isohexane (1:1); injection volume: 2 ml; column: Daicel ^Chiralcel® OX-H 5 µm, 250 x 20 mm; eluent: isohexane/ethanol 80:20; flow rate: 20 ml/min; temperature: 30°C; UV detection: 220 nm]. After separation, 29.2 mg of the first-eluting diastereomer 1 (Example 47) and 28.9 mg of the second-eluting diastereomer 2 (Example 48) 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 )

Analytical chiral HPLC: R _t = 1.49 min, de = 100% [column: Daicel Chiralpack OX-3 3µm, 50 x 4.6 mm; eluent: isohexane/ethanol 80:20; flow rate: 1 ml/min; temperature: 30°C; UV detection: 220 nm].

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 )

Analytical chiral HPLC: R _t = 2.02 min, de = 99.8% [column: Daicel Chiralpack OX-3 3µm, 50 x 4.6 mm; eluent: isohexane/ethanol 80:20; flow rate: 1 ml/min; temperature: 30°C; UV detection: 220 nm].

Example 49

5-(4-Chlorophenyl)-2-({1-(2,5-dichlorophenyl)-5-[(1 RS )-1-hydroxyethyl]-1 H -1,2,4-triazol-3-yl}methyl)-4-[(2 S )-3,3,3-trifluoro-2-hydroxypropyl]-2,4-dihydro-3 H -1,2,4-triazol-3-one ( diastereoisomer mixture )

To a solution of 5-(4-chlorophenyl)-2-({5-[(1 RS )-1-hydroxyethyl]-1 H -1,2,4-triazol-3-yl}methyl)-4-[(2 S )-3,3,3-trifluoro-2-hydroxypropyl]-2,4-dihydro-3 H -1,2,4-triazol-3-one (400 mg, 0.92 mmol) in pyridine (12 ml) was 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° C. for 2 h and then stirred at room temperature for 3 days. Then, due to incomplete conversion, additional boronic acid (100 mg, 0.52 mmol) was added. The reaction mixture was stirred at room temperature for an additional 6 days. During this time, another portion of boric acid (100 mg, 0.52 mmol) was added. The resulting reaction mixture was then concentrated in vacuo, 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] to obtain the title compound (52.8 mg, 0.09 mmol, 97% purity) as a mixture of diastereomers (yield 9.6%).

The two diastereomers were separated by preparative chiral HPLC [sample preparation: 50 mg dissolved in 10 ml of methanol; injection volume: 0.5 ml; column: Daicel ^Chiralcel® OX-H 5 µm, 250 x 20 mm; eluent: carbon dioxide/methanol 82:18; flow rate: 80 ml/min; temperature: 40°C; UV detection: 210 nm]. After separation, 20.3 mg of the first-eluting diastereomer 1 (Example 50) and 24.1 mg of the second-eluting diastereomer 2 (Example 51) 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 )

Analytical chiral HPLC (SFC): R _t = 2.87 min, de = 100% [column: Daicel Chiralcel ^® OX-3, 250 x 4 mm; eluent: carbon dioxide/methanol (5% → 60%); flow rate: 3 ml/min; UV detection: 220 nm].

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 )

Analytical chiral HPLC (SFC): R _t = 3.11 min, de = 100% [column: Daicel Chiralcel ^® OX-3, 250 x 4 mm; eluent: carbon dioxide/methanol (5% → 60%); flow rate: 3 ml/min; UV detection: 220 nm].

Example 52

2-({1-(3-chloro-2-fluorophenyl)-5-[(1 RS )-1-hydroxyethyl]-1 H -1,2,4-triazol-3-yl}methyl)-5-(4-chlorophenyl)-4-[(2 S )-3,3,3-trifluoro-2-hydroxypropyl]-2,4-dihydro-3 H -1,2,4-triazol-3-one ( diastereoisomer mixture)

To a solution of 5-(4-chlorophenyl)-2-({5-[(1 RS )-1-hydroxyethyl]-1 H -1,2,4-triazol-3-yl}methyl)-4-[(2 S )-3,3,3-trifluoro-2-hydroxypropyl]-2,4-dihydro-3 H -1,2,4-triazol-3-one (400 mg, 0.92 mmol) in pyridine (12 ml) was 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°C for 2 h and then stirred at room temperature for 12 days, during which time additional boronic acid (322.31 mg, 1.85 mmol total) was added portionwise daily. The resulting reaction mixture was then concentrated in vacuo, 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] to obtain the title compound (15.9 mg, 0.03 mmol, 97% purity) as a mixture of diastereomers (yield 3%).

The two diastereomers were separated by preparative chiral HPLC [sample preparation: 14 mg dissolved in 1 ml of ethanol/isohexane (1:1); injection volume: 1 ml; column: Daicel ^Chiralcel® OX-H 5 µm, 250 x 20 mm; eluent: isohexane/ethanol 80:20; flow rate: 15 ml/min; temperature: 25°C; UV detection: 220 nm]. After separation, 6 mg of the first-eluting diastereomer 1 (Example 53) and 6 mg of the second-eluting diastereomer 2 (Example 54) were isolated.

Example 53

2-{[1-(3-chloro-2-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-dihydro- 3H -1,2,4-triazol-3-one ( Diastereomer 1 )

Analytical chiral HPLC: R _t = 5.49 min, de = 100% [column: Daicel Chiralcel ^® OX-H 5µm, 250 x 4.6 mm; eluent: isohexane/ethanol 70:30 + 0.2% TFA and 1% water; flow rate: 1 ml/min; temperature: 40°C; UV detection: 220 nm].

Example 54

2-{[1-(3-chloro-2-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-dihydro- 3H -1,2,4-triazol-3-one ( diastereomer 2 )

Analytical chiral HPLC: R _t = 6.16 min, de = 100% [column: Daicel Chiralcel ^® OX-H 5µm, 250 x 4.6 mm; eluent: isohexane/ethanol 70:30 + 0.2% TFA and 1% water; flow rate: 1 ml/min; temperature: 40°C; UV detection: 220 nm].

Example 55

5-(4-Chlorophenyl)-2-({1-[3-(difluoromethoxy)phenyl]-5-[(1 RS )-1-hydroxyethyl]-1 H -1,2,4-triazol-3-yl}methyl)-4-[(2 S )-3,3,3-trifluoro-2-hydroxypropyl]-2,4-dihydro-3 H -1,2,4-triazol-3-one ( diastereoisomer mixture )

To a solution of 5-(4-chlorophenyl)-2-({5-[(1 RS )-1-hydroxyethyl]-1 H -1,2,4-triazol-3-yl}methyl)-4-[(2 S )-3,3,3-trifluoro-2-hydroxypropyl]-2,4-dihydro-3 H -1,2,4-triazol-3-one (400 mg, 0.92 mmol) in pyridine (12 ml) was 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° C. for 2 h and then stirred at room temperature for 6 days. Then, due to incomplete conversion, additional boronic acid (100 mg, 0.53 mmol) was added. The reaction mixture was stirred at room temperature for an additional 2 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 target compound (60.3 mg, 0.10 mmol) was obtained as a mixture of diastereomers (yield 11.4%).

The two diastereomers were separated by preparative chiral HPLC [sample preparation: 58 mg dissolved in 2 ml of ethanol; injection volume: 0.7 ml; column: Daicel ^Chiralcel® OX-H 5 µm, 250 x 20 mm; eluent: isohexane/ethanol 80:20; flow rate: 15 ml/min; temperature: 35°C; UV detection: 220 nm]. After separation, 20.7 mg of the first-eluting diastereomer 1 (Example 56) and 17.7 mg of the second-eluting diastereomer 2 (Example 57) 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 )

Analytical chiral HPLC: R _t = 5.57 min, de = 98.7% [column: Daicel Chiralcel ^® OX-H5, 250 x 4.6 mm; eluent: isohexane/ethanol 70:30 + 0.2% TFA and 1% water; flow rate: 1 ml/min; temperature: 35°C; UV detection: 220 nm].

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 )

Analytical chiral HPLC: R _t = 6.70 min, de = 100% [column: Daicel Chiralcel ^® OX-H 5,250 x 4.6 mm; eluent: isohexane/ethanol 70:30 + 0.2% TFA and 1% water; flow rate: 1 ml/min; temperature: 35°C; UV detection: 220 nm].

Example 58

2-({1-(2-chloro-5-fluorophenyl)-5-[(1 RS )-1-hydroxyethyl]-1 H -1,2,4-triazol-3-yl}methyl)-5-(4-chlorophenyl)-4-[(2 S )-3,3,3-trifluoro-2-hydroxypropyl]-2,4-dihydro-3 H -1,2,4-triazol-3-one ( diastereoisomer mixture)

To a solution of 5-(4-chlorophenyl)-2-({5-[(1 RS )-1-hydroxyethyl]-1 H -1,2,4-triazol-3-yl}methyl)-4-[(2 S )-3,3,3-trifluoro-2-hydroxypropyl]-2,4-dihydro-3 H -1,2,4-triazol-3-one (400 mg, 0.92 mmol) in pyridine (12 ml) was 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°C for 2 h and then stirred at room temperature for 10 days. During this time, additional boronic acid was added portionwise daily (total 322.31 mg, 1.85 mmol). 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] to afford the title compound (61 mg, 0.11 mmol, 98% purity) as a mixture of diastereomers (yield 11.5%).

The two diastereomers were separated by preparative chiral HPLC [sample preparation: 58 mg dissolved in 3 ml of ethanol/isohexane (2:1); injection volume: 1 ml; column: Daicel ^Chiralcel® OX-H 5 µm, 250 x 20 mm; eluent: isohexane/ethanol 80:20; flow rate: 15 ml/min; temperature: 25°C; UV detection: 220 nm]. After separation, 25 mg of the first-eluting diastereomer 1 (Example 59) and 25 mg of the second-eluting diastereomer 2 (Example 60) were isolated.

Example 59

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-dihydro- 3H -1,2,4-triazol-3-one ( Diastereomer 1 )

Analytical chiral HPLC: R _t = 5.43 min, de = 100% [column: Daicel Chiralcel ^® OX-H 5µm, 250 x 4.6 mm; eluent: isohexane/ethanol 70:30 + 0.2% TFA and 1% water; flow rate: 1 ml/min; temperature: 40°C; UV detection: 220 nm].

Example 60

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-dihydro- 3H -1,2,4-triazol-3-one ( Diastereomer 2 )

Analytical chiral HPLC: R _t = 6.11 min, de = 100% [column: Daicel Chiralcel ^® OX-H 5µm, 250 x 4.6 mm; eluent: isohexane/ethanol 70:30 + 0.2% TFA and 1% water; flow rate: 1 ml/min; temperature: 40°C; UV detection: 220 nm].

Example 61

5-(4-Chlorophenyl)-2-({1-(2,3-dichlorophenyl)-5-[(1 RS )-1-hydroxyethyl]-1 H -1,2,4-triazol-3-yl}methyl)-4-[(2 S )-3,3,3-trifluoro-2-hydroxypropyl]-2,4-dihydro-3 H -1,2,4-triazol-3-one ( diastereoisomer mixture )

To a solution of 5-(4-chlorophenyl)-2-({5-[(1 RS )-1-hydroxyethyl]-1 H -1,2,4-triazol-3-yl}methyl)-4-[(2 S )-3,3,3-trifluoro-2-hydroxypropyl]-2,4-dihydro-3 H -1,2,4-triazol-3-one (500 mg, 0.92 mmol, 80% purity) in pyridine (12 ml) was 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° C. for 1 h and then stirred at room temperature for 24 h. Then, due to incomplete conversion, additional boronic acid (80 mg, 0.42 mmol) was added. The reaction mixture was stirred at room temperature for an additional 5 days. During this period, 2 more portions of boric acid (total 160 mg, 0.84 mmol) were added. The resulting reaction mixture was then concentrated in vacuo, 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] to obtain the target compound (148 mg, 0.25 mmol, purity 97.3%) (yield 27%) as a mixture of diastereomers.

The two diastereomers were separated by preparative chiral HPLC (SFC) [sample preparation: dissolving 141 mg in 18 ml of methanol; injection volume: 0.3 ml; column: Daicel ^Chiralcel® OX-H 5 µm, 250 x 20 mm; eluent: carbon dioxide/methanol 70:30; flow rate: 80 ml/min; temperature: 40°C; UV detection: 210 nm]. After separation, 58.5 mg of the first-eluting diastereomer 1 (Example 62) and 53 mg of the second-eluting diastereomer 2 (Example 63) 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 )

Analytical chiral HPLC (SFC): R _t = 3.09 min, de = 100% [column: Daicel Chiralcel ^® OX-3 250 x 4 mm; eluent: carbon dioxide/methanol (5% → 60%); flow rate: 3 ml/min; UV detection: 220 nm].

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 )

Analytical chiral HPLC (SFC): R _t = 3.38 min, de = 100% [column: Daicel Chiralcel ^® OX-3 250 x 4 mm; eluent: carbon dioxide/methanol (5% → 60%); flow rate: 3 ml/min; UV detection: 220 nm].

Example 64

5-(4-Chlorophenyl)-2-({1-(2,3-difluorophenyl)-5-[(1 RS )-1-hydroxyethyl]-1 H -1,2,4-triazol-3-yl}methyl)-4-[(2 S )-3,3,3-trifluoro-2-hydroxypropyl]-2,4-dihydro-3 H -1,2,4-triazol-3-one ( diastereoisomer mixture )

To a solution of 5-(4-chlorophenyl)-2-({5-[(1 RS )-1-hydroxyethyl] -1H- 1,2,4-triazol-3-yl}methyl)-4-[(2 S )-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) was 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° C. for 1 h and then stirred at room temperature for 24 h. Then, due to incomplete conversion, additional boronic acid (80 mg, 0.51 mmol) was added. The reaction mixture was stirred at room temperature for an additional 5 days. During this time, 5 more portions of boric acid (400 mg, 2.54 mmol in total) were added. The resulting reaction mixture was then concentrated in vacuo, 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] to obtain the title compound (44 mg, 0.08 mmol) as a mixture of diastereomers (yield 8.1%).

The two diastereomers were separated by preparative chiral HPLC [sample preparation: dissolve 40 mg in 1 ml of ethanol; injection volume: 0.5 ml; column: Daicel ^Chiralcel® OX-H 5 µm, 250 x 20 mm; eluent: isohexane/ethanol 80:20; flow rate: 15 ml/min; temperature: 35°C; UV detection: 220 nm]. After separation, 18 mg of the first-eluting diastereomer 1 (Example 65) and 16 mg of the second-eluting diastereomer 2 (Example 66) 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 )

Analytical chiral HPLC: R _t = 5.74 min, de = 100% [column: Daicel Chiralcel ^® OX-H 5µm, 250 x 4.6 mm; eluent: isohexane/ethanol 70:30 + 0.2% TFA and 1% water; flow rate: 1 ml/min; temperature: 35°C; UV detection: 220 nm].

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 )

Analytical chiral HPLC: R _t = 6.59 min, de = 99.2% [column: Daicel Chiralcel ^® OX-H 5 µm, 250 x 4.6 mm; eluent: isohexane/ethanol 70:30 + 0.2% TFA and 1% water; flow rate: 1 ml/min; temperature: 35°C; UV detection: 220 nm].

Example 67

2-{[1-(2-chloro-3-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-dihydro- 3H -1,2,4-triazol-3-one ( Diastereomer 1 )

To a solution of 5-(4-chlorophenyl)-2-({5-[(1 RS )-1-hydroxyethyl]-1 H -1,2,4-triazol-3-yl}methyl)-4-[(2 S )-3,3,3-trifluoro-2-hydroxypropyl]-2,4-dihydro-3 H -1,2,4-triazol-3-one (500 mg, 0.92 mmol, 80% purity) in pyridine (12 ml) was 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° C. for 1 h and then stirred at room temperature for 24 h. Then, due to incomplete conversion, additional boronic acid (75 mg, 0.43 mmol) was added. The reaction mixture was stirred at room temperature for an additional 6 days. During this time, 5 more portions of boric acid (375 mg, 2.15 mmol in total) were added. The resulting reaction mixture was then concentrated in vacuo, 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 target compound was isolated as a mixture of diastereomers still containing some impurities.

Further purification by preparative chiral HPLC afforded two pure, separated diastereomers [sample preparation: 90 mg dissolved in 3 ml ethanol; injection volume: 0.3 ml; column: Daicel ^Chiralcel® OX-H 5 µm, 250 x 20 mm; eluent: isohexane/ethanol 80:20; flow rate: 15 ml/min; temperature: 35°C; UV detection: 220 nm]. After separation, 20 mg of the first-eluting diastereomer 1 (Example 67) and 21 mg of the second-eluting diastereomer 2 (Example 68) were isolated.

Analytical chiral HPLC: R _t = 6.22 min, de = 100% [column: Daicel Chiralcel ^® OX-H 5µm, 250 x 4.6 mm; eluent: isohexane/ethanol 70:30 + 0.2% TFA and 1% water; flow rate: 1 ml/min; temperature: 35°C; UV detection: 220 nm].

Example 68

2-{[1-(2-chloro-3-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-dihydro- 3H -1,2,4-triazol-3-one ( diastereomer 2 )

Analytical chiral HPLC: R _t = 7.94 min, de = 100% [column: Daicel Chiralcel ^® OX-H 5µm, 250 x 4.6 mm; eluent: isohexane/ethanol 70:30 + 0.2% TFA and 1% water; flow rate: 1 ml/min; temperature: 35°C; UV detection: 220 nm].

Example 69

5-(4-Chlorophenyl)-2-({5-[(1 RS )-1-hydroxyethyl]-1-(2-methylphenyl)-1 H -1,2,4-triazol-3-yl}methyl)-4-[(2 S )-3,3,3-trifluoro-2-hydroxypropyl]-2,4-dihydro-3 H -1,2,4-triazol-3-one ( diastereoisomer mixture )

To a solution of 5-(4-chlorophenyl)-2-({5-[(1 RS )-1-hydroxyethyl]-1 H -1,2,4-triazol-3-yl}methyl)-4-[(2 S )-3,3,3-trifluoro-2-hydroxypropyl]-2,4-dihydro-3 H -1,2,4-triazol-3-one (400 mg, 0.92 mmol) in pyridine (12 ml) was 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, then, due to incomplete conversion, additional boronic acid (62.8 mg, 0.46 mmol, 0.5 eq.) was added. After stirring for an additional 2 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] to afford the title compound (100 mg, 0.17 mmol) as a mixture of diastereomers (yield 17.2%, purity 90%).

The two diastereomers were separated by preparative chiral HPLC [sample preparation: 98 mg dissolved in 2 ml of ethanol/isohexane (1:1); injection volume: 1 ml; column: Daicel ^Chiralcel® OX-H 5 µm, 250 x 20 mm; eluent: isohexane/ethanol 75:25; flow rate: 15 ml/min; temperature: 30°C; UV detection: 220 nm]. After separation, 37 mg of the first-eluting diastereomer 1 (Example 70) and 39 mg of the second-eluting diastereomer 2 (Example 71) were isolated.

Example 70

5-(4-Chlorophenyl)-2-{[5-(1-hydroxyethyl)-1-(2-methylphenyl) -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 )

Analytical chiral HPLC: R _t = 7.65 min, de = 100% [column: LUX Cellulose-4, 5 µm, 250 x 4.6 mm; eluent: isohexane/ethanol 70:30; flow rate: 1 ml/min; temperature: 40°C; UV detection: 220 nm].

Example 71

5-(4-Chlorophenyl)-2-{[5-(1-hydroxyethyl)-1-(2-methylphenyl) -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 )

Analytical chiral HPLC: R _t = 10.27 min, de = 100% [column: LUX Cellulose-4, 5 µm, 250 x 4.6 mm; eluent: isohexane/ethanol 70:30; flow rate: 1 ml/min; temperature: 40°C; UV detection: 220 nm].

Example 72

5-(4-Chlorophenyl)-2-({5-[(1 RS )-1-hydroxyethyl]-1-[2-(trifluoromethyl)phenyl]-1 H -1,2,4-triazol-3-yl}methyl)-4-[(2 S )-3,3,3-trifluoro-2-hydroxypropyl]-2,4-dihydro-3 H -1,2,4-triazol-3-one ( diastereoisomer mixture )

To a solution of 5-(4-chlorophenyl)-2-({5-[(1 RS )-1-hydroxyethyl]-1 H -1,2,4-triazol-3-yl}methyl)-4-[(2 S )-3,3,3-trifluoro-2-hydroxypropyl]-2,4-dihydro-3 H -1,2,4-triazol-3-one (600 mg, 1.11 mmol, 80% purity) in pyridine (14.5 ml) was 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° C. for 2 h and then stirred at room temperature for 5 days. Due to incomplete conversion, additional boronic acid (105 mg, 0.55 mmol, 0.5 eq.) was added. After stirring overnight at room temperature, 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 target compound (80 mg) was obtained as a mixture of diastereomers (yield 12.4%).

The two diastereomers were separated by preparative chiral HPLC [sample preparation: 78 mg dissolved in 2 ml of ethanol/isohexane (1:1); injection volume: 1 ml; column: Daicel ^Chiralcel® OX-H 5 µm, 250 x 20 mm; eluent: isohexane/ethanol 75:25; flow rate: 15 ml/min; temperature: 30°C; UV detection: 220 nm]. After separation, 34 mg of the first-eluting diastereomer 1 (Example 73) and 30 mg of the second-eluting diastereomer 2 (Example 74) were isolated.

Example 73

5-(4-Chlorophenyl)-2-({5-(1-hydroxyethyl)-1-[2-(trifluoromethyl)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 ( Diastereomer 1 )

Analytical chiral HPLC: R _t = 6.16 min, de = 100% [column: LUX Cellulose-4, 5 µm, 250 x 4.6 mm; eluent: isohexane/ethanol 70:30; flow rate: 1 ml/min; temperature: 40°C; UV detection: 220 nm].

Example 74

5-(4-Chlorophenyl)-2-({5-(1-hydroxyethyl)-1-[2-(trifluoromethyl)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 ( Diastereomer 2 )

Analytical chiral HPLC: R _t = 8.67 min, de = 100% [column: LUX Cellulose-4, 5 µm, 250 x 4.6 mm; eluent: isohexane/ethanol 70:30; flow rate: 1 ml/min; temperature: 40°C; UV detection: 220 nm].

Example 75

5-(4-Chlorophenyl)-2-({1-(3-fluorophenyl)-5-[(1 RS )-1-hydroxyethyl]-1 H -1,2,4-triazol-3-yl}methyl)-4-[(2 S )-3,3,3-trifluoro-2-hydroxypropyl]-2,4-dihydro-3 H -1,2,4-triazol-3-one ( diastereoisomer mixture )

To a solution of 5-(4-chlorophenyl)-2-({5-[(1 RS )-1-hydroxyethyl]-1 H -1,2,4-triazol-3-yl}methyl)-4-[(2 S )-3,3,3-trifluoro-2-hydroxypropyl]-2,4-dihydro-3 H -1,2,4-triazol-3-one (430 mg, 0.795 mmol, 80% purity) in pyridine (10 ml) was 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° C. for 2 h and then stirred at room temperature for 5 days. Then, due to incomplete conversion, additional boronic acid (55.6 mg, 0.40 mmol) was added. The reaction mixture was heated to 60° C. for 2 h and then stirred 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] to obtain the target compound (100 mg, 0.19 mmol) (yield 23.9%) as a mixture of diastereomers.

The two diastereomers were separated by preparative chiral HPLC [sample preparation: 97 mg dissolved in 4 ml of ethanol/isohexane (1:1); injection volume: 1 ml; column: Daicel ^Chiralcel® OX-H 5 µm, 250 x 20 mm; eluent: isohexane/ethanol 80:20; flow rate: 15 ml/min; temperature: 30°C; UV detection: 220 nm]. After separation, 36 mg of the first-eluting (1 S )-diastereomer (Example 76) and 40 mg of the second-eluting (1 R )-diastereomer (Example 77) were isolated.

Example 76

5-(4-chlorophenyl)-2-({1-(3-fluorophenyl)-5-[(1 S )-1-hydroxyethyl]-1 H -1,2,4-triazol-3-yl}methyl)-4-[(2 S )-3,3,3-trifluoro-2-hydroxypropyl]-2,4-dihydro-3 H -1,2,4-triazol-3-one

Analytical chiral HPLC: R _t = 9.71 min, de = 100% [column: LUX Cellulose-4, 5 µm, 250 x 4.6 mm; eluent: isohexane/ethanol 80:20; flow rate: 1 ml/min; temperature: 40°C; UV detection: 220 nm].

The absolute stereochemistry of this compound was determined by additionally performing the same reaction starting from the enantiomerically pure diastereoisomer 5-(4-chlorophenyl)-2-({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 (Example 5A) and comparing the respective products by analytical chiral HPLC.

Example 77

5-(4-chlorophenyl)-2-({1-(3-fluorophenyl)-5-[(1 R )-1-hydroxyethyl]-1 H -1,2,4-triazol-3-yl}methyl)-4-[(2 S )-3,3,3-trifluoro-2-hydroxypropyl]-2,4-dihydro-3 H -1,2,4-triazol-3-one

Analytical chiral HPLC: R _t = 13.60 min, de = 100% [column: LUX Cellulose-4, 5 µm, 250 x 4.6 mm; eluent: isohexane/ethanol 80:20; flow rate: 1 ml/min; temperature: 40°C; UV detection: 220 nm].

The absolute stereochemistry of this compound was determined by additionally performing the identical reaction starting from the enantiomerically pure diastereoisomer 5-(4-chlorophenyl)-2-({5-[(1 R )-1-hydroxyethyl]-1 H -1,2,4-triazol-3-yl}methyl)-4-[(2 S )-3,3,3-trifluoro-2-hydroxypropyl]-2,4-dihydro-3 H -1,2,4-triazol-3-one (Example 6A) and comparing the respective products by analytical chiral HPLC.

Example 78

5-(4-Chlorophenyl)-2-({1-(3-chlorophenyl)-5-[(1 RS )-1-hydroxyethyl]-1 H -1,2,4-triazol-3-yl}methyl)-4-[(2 S )-3,3,3-trifluoro-2-hydroxypropyl]-2,4-dihydro-3 H -1,2,4-triazol-3-one ( diastereoisomer mixture )

To a solution of 5-(4-chlorophenyl)-2-({5-[(1 RS )-1-hydroxyethyl]-1 H -1,2,4-triazol-3-yl}methyl)-4-[(2 S )-3,3,3-trifluoro-2-hydroxypropyl]-2,4-dihydro-3 H -1,2,4-triazol-3-one (430 mg, 0.795 mmol, 80% purity) in pyridine (10 ml) was 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° C. for 2 h and then stirred at room temperature for 5 days. Then, due to incomplete conversion, additional boronic acid (62.1 mg, 0.40 mmol) was added. The reaction mixture was heated to 60° C. for 2 h and then stirred 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 target compound (130 mg, 0.24 mmol) was obtained as a mixture of diastereomers (yield 30.1%).

The two diastereomers were separated by preparative chiral HPLC [sample preparation: 128 mg dissolved in 4 ml of ethanol/isohexane (1:1); injection volume: 1 ml; column: Daicel ^Chiralcel® OX-H 5 µm, 250 x 20 mm; eluent: isohexane/ethanol 80:20; flow rate: 15 ml/min; temperature: 30°C; UV detection: 220 nm]. After separation, 52 mg of the first-eluting (1 S )-diastereomer (Example 79) and 49 mg of the second-eluting (1 R )-diastereomer (Example 80) were isolated.

Example 79

5-(4-chlorophenyl)-2-({1-(3-chlorophenyl)-5-[(1 S )-1-hydroxyethyl]-1 H -1,2,4-triazol-3-yl}methyl)-4-[(2 S )-3,3,3-trifluoro-2-hydroxypropyl]-2,4-dihydro-3 H -1,2,4-triazol-3-one

Analytical chiral HPLC: R _t = 9.96 min, de = 100% [column: LUX Cellulose-4, 5 µm, 250 x 4.6 mm; eluent: isohexane/ethanol 80:20; flow rate: 1 ml/min; temperature: 35°C; UV detection: 220 nm].

The absolute stereochemistry of this compound was determined by additionally performing the same reaction starting from the enantiomerically pure diastereoisomer 5-(4-chlorophenyl)-2-({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 (Example 5A) and comparing the respective products by analytical chiral HPLC.

Example 80

5-(4-chlorophenyl)-2-({1-(3-chlorophenyl)-5-[(1 R )-1-hydroxyethyl]-1 H -1,2,4-triazol-3-yl}methyl)-4-[(2 S )-3,3,3-trifluoro-2-hydroxypropyl]-2,4-dihydro-3 H -1,2,4-triazol-3-one

Analytical chiral HPLC: R _t = 14.41 min, de = 100% [column: LUX Cellulose-4, 5 µm, 250 x 4.6 mm; eluent: isohexane/ethanol 80:20; flow rate: 1 ml/min; temperature: 35°C; UV detection: 220 nm].

The absolute stereochemistry of this compound was determined by additionally performing the identical reaction starting from the enantiomerically pure diastereoisomer 5-(4-chlorophenyl)-2-({5-[(1 R )-1-hydroxyethyl]-1 H -1,2,4-triazol-3-yl}methyl)-4-[(2 S )-3,3,3-trifluoro-2-hydroxypropyl]-2,4-dihydro-3 H -1,2,4-triazol-3-one (Example 6A) and comparing the respective products by analytical chiral HPLC.

Example 81

5-(4-Chlorophenyl)-2-({1-(2-chlorophenyl)-5-[(1 RS )-1-hydroxyethyl]-1 H -1,2,4-triazol-3-yl}methyl)-4-[(2 S )-3,3,3-trifluoro-2-hydroxypropyl]-2,4-dihydro-3 H -1,2,4-triazol-3-one ( diastereoisomer mixture )

To a solution of 5-(4-chlorophenyl)-2-({5-[(1 RS )-1-hydroxyethyl]-1 H -1,2,4-triazol-3-yl}methyl)-4-[(2 S )-3,3,3-trifluoro-2-hydroxypropyl]-2,4-dihydro-3 H -1,2,4-triazol-3-one (2.10 g, 3.88 mmol, 80% purity) in pyridine (50 ml) was 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° C. for 1 h and then stirred at room temperature for 5 days. Then, due to incomplete conversion, additional boronic acid (303 mg, 1.94 mmol) was added. After stirring at room temperature for another 2 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 target compound (580 mg, 1.01 mmol, purity 95%) was obtained as a mixture of diastereomers (yield 26.1%).

The two diastereomers were separated by preparative chiral HPLC (SFC) [sample preparation: 575 mg dissolved in 35 ml of methanol; injection volume: 0.4 ml; column: Daicel ^Chiralcel® OX-H 5 µm, 250 x 20 mm; eluent: carbon dioxide/methanol 70:30; flow rate: 80 ml/min; temperature: 40°C; UV detection: 210 nm]. After separation, 206 mg of the first-eluting (1 S )-diastereomer (Example 82) and 189 mg of the second-eluting (1 R )-diastereomer (Example 83) were isolated.

Example 82

5-(4-chlorophenyl)-2-({1-(2-chlorophenyl)-5-[(1 S )-1-hydroxyethyl]-1 H -1,2,4-triazol-3-yl}methyl)-4-[(2 S )-3,3,3-trifluoro-2-hydroxypropyl]-2,4-dihydro-3 H -1,2,4-triazol-3-one

Analytical chiral HPLC: R _t = 8.34 min, de = 100% [column: LUX Cellulose-4, 5 µm, 250 x 4.6 mm; eluent: isohexane/ethanol 70:30; flow rate: 1 ml/min; temperature: 40°C; UV detection: 220 nm].

The absolute stereochemistry of this compound was determined by additionally performing the same reaction starting from the enantiomerically pure diastereoisomer 5-(4-chlorophenyl)-2-({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 (Example 5A) and comparing the respective products by analytical chiral HPLC.

Example 83

5-(4-chlorophenyl)-2-({1-(2-chlorophenyl)-5-[(1 R )-1-hydroxyethyl]-1 H -1,2,4-triazol-3-yl}methyl)-4-[(2 S )-3,3,3-trifluoro-2-hydroxypropyl]-2,4-dihydro-3 H -1,2,4-triazol-3-one

Analytical chiral HPLC: R _t = 11.88 min, de = 98.1% [column: LUX Cellulose-4, 5 µm, 250 x 4.6 mm; eluent: isohexane/ethanol 70:30; flow rate: 1 ml/min; temperature: 40°C; UV detection: 220 nm].

The absolute stereochemistry of this compound was determined by additionally performing the identical reaction starting from the enantiomerically pure diastereoisomer 5-(4-chlorophenyl)-2-({5-[(1 R )-1-hydroxyethyl]-1 H -1,2,4-triazol-3-yl}methyl)-4-[(2 S )-3,3,3-trifluoro-2-hydroxypropyl]-2,4-dihydro-3 H -1,2,4-triazol-3-one (Example 6A) and comparing the respective products by analytical chiral HPLC.

B. Evaluation of biological activity

Abbreviations and Acronyms:

Acc. No.

AVP Arginine vasopressin

B _max maximum ligand binding capacity

BSA bovine serum albumin

cAMP cyclic adenosine monophosphate

Cat. No.

cDNA complementary deoxyribonucleic acid

CHO Chinese Hamster Ovary

CRE cAMP response element

Ct cycle threshold

DMEM/F12 Dulbecco's Modified Eagle's Medium/Ham's F12 Medium (1:1)

DNA deoxyribonucleic acid

DTT dithiothreitol

_EC50 half effective concentration

EDTA Ethylenediaminetetraacetic acid

FAM carboxyfluorescein succinimidyl ester

f.c. Final concentration

FCS fetal calf serum

HEPES 4-(2-Hydroxyethyl)piperazine-1-ethanesulfonic acid

_IC50 half-maximal inhibitory concentration

_Kd dissociation constant

Dissociation constant of K _i inhibitor

mRNA messenger ribonucleic acid

PBS Phosphate-buffered saline

p.o. Oral

RNA ribonucleic acid

RTPCR real-time polymerase chain reaction

SPA Scintillation Proximity Analysis

TAMRA Carboxytetramethylrhodamine

TRIS 2-amino-2-hydroxymethylpropane-1,3-diol.

The activity of the compounds of the present invention can be verified by in vitro, ex vivo and in vivo tests well known in the art. For example, in order to verify the activity of the compounds of the present invention, the following tests can be used.

B-1. In vitro cell assay for measuring vasopressin receptor activity

Recombinant cell lines were used to identify agonists and antagonists of the human, rat, and dog Via and V2 vasopressin receptors and to quantify the activity of the compounds of the invention. These cell lines were originally derived from hamster ovary epithelial cells (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 Via or V2 receptors. In the case of _Gαq -coupled V1a receptors, cells were also stably transfected with modified forms of the calcium-sensitive photoproteins aequorin (human and rat V1a) or obelin (dog V1a), which, after reconstitution with the cofactor coelenterazine, emit light when free calcium concentrations increase [Rizzuto R, Simpson AW, Brini M, Pozzan T, Nature 358 , 325-327 (1992); Illarionov BA, Bondar VS, Illarionova VA, Vysotski ES, Gene 153 (2), 273-274 (1995)]. The resulting vasopressin receptor cells responded to stimulation of the recombinantly expressed V1a receptor by intracellular release of calcium ions, which could be quantified by the luminescence of the resulting photoprotein. Gs _- coupled V2 receptors were stably transfected into a cell line expressing the gene for firefly luciferase under the control of a CRE-driven promoter. Activation of the V2 receptor induces activation of the CRE-driven promoter through an increase in cAMP, thereby inducing expression of firefly luciferase. Light emitted by the photoproteins of the V1a cell line and by the firefly luciferase of the V2 cell line corresponds to activation or inhibition of the respective vasopressin receptors. Bioluminescence of the cell lines can be measured 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:

One day before the experiment, the cells were plated in culture medium (DMEM/F12, 2% FCS, 2 mM glutamine, 10 mM HEPES, 5 µg/ml coelenterazine) in 384-well microtiter plates and maintained in a cell culture incubator (96% humidity, 5% v/v CO ₂ , 37°C). On the day of the experiment, the test compound was placed in the wells of the microtiter plate at various concentrations for 10 minutes, followed by the addition of the agonist [Arg ⁸ ]-vasopressin at the EC ₅₀ concentration. The resulting light signal was immediately measured in a photometer.

Vasopressin V2 receptor cell lines:

One day before the assay, cells were plated in 384-well microtiter plates in culture medium (DMEM/F12, 2% FCS, 2 mM glutamine, 10 mM HEPES) and maintained in a cell culture incubator (96% humidity, 5% v/v CO ₂ , 37°C). On the day of the assay, varying concentrations of the test compound were added to the wells along with the EC ₅₀ concentration of the agonist [Arg ⁸ ]-vasopressin, and the plates were incubated in a cell culture incubator for 3 hours. After the addition of the cell lysis reagent Triton ^™ and the substrate luciferin, luminescence of firefly luciferase was measured in a luminometer.

Table 1A below lists individual _IC50 values for compounds of the invention (including diastereomeric mixtures as well as isolated enantiomerically pure diastereomers) obtained from cell lines transfected with the human Via or V2 receptor.

The _IC50 data listed in Table 1A indicate that the compounds of the present invention act as very potent dual antagonists of the vasopressin V1a and V2 receptors.

For comparative purposes, selected phenyltriazole and imidazole derivatives considered to represent the closest prior art ( cf. International Patent Application WO 2011/104322-A1 and the example compounds described therein) were also tested in the cellular V1a and V2 assays described above. The _IC50 values of these compounds, obtained using cell lines transfected with the human V1a or V2 receptor, are listed in Table 1B below.

B-2. Radioactive incorporation assay

_IC50 and _K values were determined in radioactive binding competition SPA assays using membrane fractions from recombinant CHO cell lines expressing human, rat, or dog vasopressin V1a and V2 receptors, respectively. These cells were derived from hamster ovary epithelial cells (Chinese Hamster Ovary, CHO K1, ATCC: American Type Culture Collection, Manassas, VA 20108, USA). Furthermore, the cells were stably transfected with human, rat, or dog V1a or V2 receptors. The membrane preparations were subjected to the following radioreceptor binding competition assays.

CHO cells transfected with each vasopressin receptor were grown in appropriate amounts in T-175 flasks containing DMEM/F12, 10% FCS, 15 mM HEPES, and 1 mg/ml G418 and maintained in a cell culture incubator (96% humidity, 5% v/v _CO₂ , 37°C). After reaching appropriate cell confluency, cells were harvested from the membrane preparation. The cells were scraped into PBS and pelleted by gentle centrifugation at 200 x g for 5 minutes at room temperature. The cell pellet was resuspended in PBS and centrifuged again. This step was repeated once more, and the resulting cell pellet was snap-frozen at -80°C for 30 minutes. The frozen pellet was resuspended 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 chilled on ice for 2 minutes and the homogenization was repeated twice. The resulting homogenate was centrifuged at 500 x g for 10 minutes at 4°C. The membrane was pelleted at 4500 x g for 20 minutes at 4°C, resuspended in storage buffer (7.5 mM TRIS, 12.5 mM _MgCl₂ , 0.3 mM EDTA, 250 mM sucrose, cOmplete protease inhibitor cocktail), and homogenized at 2000 rpm for 2 seconds (Polytron PT3000, Kinematica). Protein concentration was determined using a BCA protein assay (Thermo Scientific Pierce), and the membrane preparation was stored at -80°C. On the day of use, an aliquot was thawed and briefly vortexed.

To determine the receptor binding affinity of test compounds, a SPA assay was set up as follows. _Kd and _Bmax values were determined for each membrane preparation. Based on these data, the amount of SPA beads (WGA PVT beads, PerkinElmer, 200 µg/well), the concentration of radioligand ( ^3H -AVP, PerkinElmer, 2.431 TBq/mmol, fc 1-2 x _Kd ), and the amount of each membrane preparation (10 µg protein/well) were matched to the assay volume (100 µl) in binding buffer (50 mM TRIS, 0.2% BSA) in a 96-well plate. Test compounds were diluted in binding buffer (fc ^10-4 M to ^10-12 M) and assayed. The plates were gently shaken at room temperature for 1-3 hours and incubated for a further 1-2 hours. The signal generated by the binding of 3H-AVP was measured using a beta counter (1450 MicrobetaTrilux). Based on these results, _IC50 and _K1 values of the test compounds were calculated using GraphPad Prism.

B-3. In vitro cell-based assays for detecting the regulatory effects of vasopressin V1a receptor antagonists on profibrotic genes

The H9C2 cell line (American Type Culture Collection No. CRL-1446), a cardiomyocyte type isolated from rat heart tissue, endogenously expresses the vasopressin V1A receptor AVPR1A at high copy numbers, while AVPR2 expression is undetectable. The following is a cellular assay to investigate AVPR1A receptor-dependent regulation of gene expression by inhibiting it with receptor antagonists.

H9C2 cells were seeded at a cell density of 50,000 cells/well in 2.0 ml of Opti-MEM medium (Invitrogen Corp., Carlsbad, CA, USA, catalog number 11058-021) in a 6-well microtiter plate for cell culture and maintained in a cell culture incubator (96% humidity, 8% v/v carbon dioxide, 37°C). After 24 hours, vehicle solution (negative control) and vasopressin solution ([Arg ⁸ ]-vasopressin acetate, Sigma, catalog number V9879) were added to three wells per group (in triplicate), or the test compound (dissolved in vehicle: water containing 20% v/v ethanol) and vasopressin solution. The final vasopressin concentration in the cell culture was 1 nM. The test compound solution was added to the cell culture in small volumes so that the final ethanol concentration in the cell assay did not exceed 0.03%. The 5-well plate of 100 μ l 4-well plate was used for the PCR reaction. The 5-well plate was used for the PCR reaction. The 5-well plate was used for the PCR reaction. The 5-well plate was used for the PCR reaction. The 5-well plate was used for the PCR reaction. The 5-well plate was used for the PCR reaction. The 5-well plate was used for the PCR reaction. The 5-well plate was used for the PCR reaction. The 5-well plate was used for the PCR reaction. The 5-well plate was used for the PCR reaction. The 5-well plate was used for the PCR reaction. The 5-well plate was used for the PCR reaction. The 5-well plate was used for the PCR reaction. The 5-well plate was used for the PCR reaction. RT-PCR to determine relative mRNA expression in cells from different experimental batches was performed using an Applied Biosystems ABI Prism 7700 sequence detector in 384-well microtiter plate format according to the instrument's operating instructions. Relative gene expression was expressed as delta-delta Ct values, with reference to the expression level of the ribosomal protein L-32 gene (Genbank accession number NM_013226) and a Ct threshold of 35 [Applied Biosystems, User Bulletin No. 2 ABI Prism 7700 SDS, December 11, 1997 (updated October 2001)].

B-4. In vivo test for cardiovascular effects: blood pressure measurement in anesthetized rats (vasopressin "challenge" model)

Under ketamine/xylazine/pentobarbital injection anesthesia, polyethylene tubing (PE-50; Intramedic®) pre-filled with isotonic sodium chloride solution containing heparin (500 IU/ml) was inserted into the jugular and femoral veins of male Sprague-Dawley rats (250-350 g body weight ⁾ and then fixed. Arg-vasopressin was injected via a syringe through one venous access port, and the test substance was administered via a second venous access port. To measure systolic blood pressure, a pressure catheter (Millar SPR-320 2F) was fixed in the carotid artery. The arterial catheter was connected to a pressure sensor, which transmitted its signal to a recording computer equipped with appropriate recording software. In a typical experiment, 3-4 continuous boluses of a defined amount of Arg-vasopressin (30 ng/kg) in isotonic sodium chloride solution were administered to the experimental animals at intervals of 10-15 min. When blood pressure reached the initial level again, the test substance was administered as a bolus followed by a continuous infusion in a suitable solvent. Thereafter, at defined intervals (10-15 min), the same amount of Arg-vasopressin as at the start is administered again. Based on the blood pressure values, the extent to which the test substance counteracts the pressor effect of Arg-vasopressin is determined. Control animals receive only the vehicle instead of the test substance.

After intravenous administration, the compounds of the present invention produce an inhibitory effect on the blood pressure increase caused by Arg-vasopressin, compared with a vehicle control.

B-5. In Vivo Assay for Cardiovascular Effects: Diuresis Studies in Conscious Rats Maintained in Metabolic Cages

Wistar rats (220-450 g body weight) are kept free to access feed (Altromin) and drinking water. During the experiment, in a metabolic cage suitable for these heavyweight rats, animals are individually kept free to access drinking water for 4-8 hours or until 24 hours (Tecniplast Deutschland GmbH, D-82383 Hohenpeißenberg). At the beginning of the experiment, the test substance is given to the animals using gavage with a suitable solvent volume of 1-3 ml/kg body weight. The control animals only receive the solvent. On the same day, control and substance tests are carried out in parallel. The control group and substance dosage group are each composed of 4-8 animals. During the experiment, the urine excreted by the animals is continuously collected in a receiver at the bottom of the cage. For each animal, the urine volume per unit time is measured individually, and the concentration of urine electrolytes is measured by the standard method of flame photometry. Before the experiment begins, the body weight of each animal is measured.

Following oral administration, the compounds of the invention cause increased urinary excretion compared to vehicle control administration, which is essentially based on increased water excretion (hydroprotection).

Table 2A below shows the changes in urinary excretion observed for the example compounds of the present invention at two different doses relative to the solvent control (=100%).

For comparative purposes, the diuretic effect of selected phenyltriazole and imidazole derivatives, considered to represent the closest prior art ( cf. International Patent Application WO 2011/104322-A1 and the example compounds described therein), was also tested in this study. The changes in urinary excretion observed at two different doses relative to the solvent control (= 100%) are shown in Table 2B below.

The results shown in Tables 2A and 2B indicate that the compounds of the present invention are significantly more effective in vivo: compared to the vehicle control group, at an oral dose of 3 mg/kg, the tested examples of the present invention caused an increase in urine volume of more than 3 times, and in some cases, more than 10 times, and most examples showed significant excretion-promoting activity at an oral dose of 0.3 mg/kg or 1 mg/kg. This is in contrast to the phenyltriazole and imidazole derivatives that are considered to represent the closest prior art, which are inactive at oral doses below 3 mg/kg and have low activity at an oral dose of 3 mg/kg.

B-6. In Vivo Assays for Cardiovascular Effects: Hemodynamic Studies in Anesthetized Dogs

Male beagle dogs (Marshall BioResources) weighing 10-15 kg were anesthetized with pentobarbital (30 mg/kg intravenously, Narcoren ^® , Merial, Germany) for surgical procedures and hemodynamic and functional testing. Pancuronium bromide (2 mg/animal, intravenously, Ratiopharm, Germany) was also used as a muscle relaxant. The dogs were intubated and ventilated with an oxygen/ambient air mixture (40/60%, approximately 3-4 L/min). Ventilation was maintained using a ventilator from GE Healthcare (Avance) and monitored using an analyzer (Datex-Ohmeda, GE). Anesthesia was maintained with a continuous infusion of pentobarbital (50 µg/kg/min); fentanyl was used as an analgesic (10-40 µg/kg/h). An alternative to pentobarbital is isoflurane (1-2% by volume).

In a preparatory intervention, a pacemaker was installed in the dog. 21 days before the first drug trial (i.e., the experiment began), a pacemaker (from Biotronik, ^Logos® ) was implanted in a subcutaneous pouch and contacted with the heart via a pacemaker electrode that extended into the right ventricle via the external jugular vein (using fluoroscopy). After this, all interfaces were removed and the dog was allowed to naturally wake up from anesthesia. After another 7 days (i.e., 14 days before the first drug trial), the pacemaker was activated and the heart was stimulated at a frequency of 220 beats per minute.

The actual substance testing experiments were conducted 14 and 28 days after the start of pacemaker stimulation using the following apparatus:

● Insertion of a bladder catheter, which is used to relieve the bladder and to measure urine flow;

● Connect electrocardiogram (ECG) leads to the limbs for ECG measurement;

● Fluidmedic ^® PE-300 tubing filled with sodium chloride solution was inserted into the femoral artery and connected to a pressure transducer (Braun Melsungen, Germany) for measuring systemic blood pressure;

● A Millar Tip catheter (model 350 PC, Millar Instruments, Houston, USA) was inserted via the left atrium or through a port installed in the carotid artery to measure cardiac hemodynamics;

● A Swan-Ganz catheter (CCOmbo 7.5F, Edwards, Irvine, USA) was inserted into the pulmonary artery via the jugular vein to measure cardiac output, oxygen saturation, pulmonary artery pressure, and central venous pressure;

● Placement of an intravenous catheter in the cephalic vein for infusion of pentobarbital, for fluid replacement, and for blood sampling (for determination of plasma levels of the test substance or other clinical blood values);

● Place an intravenous catheter in the saphenous vein for fentanyl infusion and for administration of the test substance;

• Vasopressin (Sigma, 4 mU/kg/min) was continuously infused, and the test compound was then administered and evaluated at various doses under this vasopressin infusion.

If necessary, the primary signal was amplified (ACQ 7700 amplifier, DataSciences Inc., Minneapolis, USA or Edwards-Vigilance-Monitor, Edwards, Irvine, USA) and subsequently input into the Ponemah system (DataSciences Inc, Minneapolis, USA) for evaluation. The signal was continuously recorded throughout the experiment and further digitally processed by the software and averaged over 30 seconds.

Although the present invention has been disclosed with reference to specific embodiments, it is apparent that those skilled in the art may design other embodiments and variations of the present invention without departing from the true spirit and scope of the invention. The claims are intended to be interpreted as including all such embodiments and equivalent variations.

C. Examples of Pharmaceutical Compositions

The pharmaceutical composition according to the present invention can be exemplified as follows.

Sterile intravenous solution:

A 5 mg/mL solution of the target compound of the present invention can be prepared using sterile water for injection and, if necessary, the pH adjusted. The solution is diluted with sterile 5% glucose to 1-2 mg/mL for administration and administered as an intravenous infusion over approximately 60 minutes.

Lyophilized powder for intravenous administration:

A sterile formulation can be prepared using ( i ) 100-1000 mg of the target compound of the present invention as a lyophilized powder, ( ii ) 32-327 mg/mL sodium citrate, and ( iii ) 300-3000 mg of dextran 40. The formulation is reconstituted with sterile injectable saline or 5% dextrose to a concentration of 10-20 mg/mL, further diluted with saline or 5% dextrose to 0.2-0.4 mg/mL, and administered as an intravenous bolus or by intravenous infusion over 15-60 minutes.

Intramuscular suspension:

The following solutions or suspensions can be prepared for intramuscular injection:

50 mg/mL desired water-insoluble compound of the present invention, 5 mg/mL sodium carboxymethylcellulose, 4 mg/mL Tween 80, 9 mg/mL sodium chloride, 9 mg/mL benzyl alcohol.

Hard shell capsules:

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 lactose, 50 mg cellulose and 6 mg magnesium stearate.

Soft gelatin capsules:

A mixture of the target compound of the present invention in a digestible oil such as soybean oil, cottonseed oil, or olive oil is prepared and injected into molten gelatin using a positive displacement pump to form soft gelatin capsules containing 100 mg of the active ingredient. The capsules are then washed and dried. A water-soluble pharmaceutical mixture can be prepared by dissolving the target compound of the present invention in a mixture of polyethylene glycol, glycerol, and sorbitol.

tablet:

A large number of tablets are prepared by conventional methods so that the dosage unit is 100 mg of the target compound of the present 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 enhance palatability, improve aesthetics and stability or delay absorption.

Claims (15)

1. A compound of formula (I) or a pharmaceutically acceptable salt thereof, in ^R1 is hydrogen or methyl, and R ^2A and R ^2B are independently selected from hydrogen, fluorine, chlorine, cyano, methyl, fluoromethyl, difluoromethyl, trifluoromethyl, ethyl, methoxy, difluoromethoxy, and trifluoromethoxy. 2. The compound of formula (I) according to claim 1, or a pharmaceutically acceptable salt thereof, wherein ^R1 is hydrogen or methyl, and ^R2A and ^R2B are independently selected from hydrogen, fluorine, chlorine, methyl and methoxy, wherein at least one of ^R2A and ^R2B is not hydrogen. 3. A compound of formula (I) according to claim 1 or 2, or a pharmaceutically acceptable salt thereof, wherein the compound is selected from: 5-(4-chlorophenyl)-2-{[1-(3-chlorophenyl)-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; 5-(4-chlorophenyl)-2-{[1-(3-fluorophenyl)-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; 5-(4-chlorophenyl)-2-{[5-(hydroxymethyl)-1-(2-methylphenyl)-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; 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-[(1R)-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-[(1S)-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-[(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-[(1R)-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-[(1S)-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; 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-dihydro-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-dihydro-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-dihydro-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-dihydro-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-dihydro-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-dihydro-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-dihydro-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-dihydro-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-dihydro-3H-1,2,4-triazol-3-one. 4. A compound of formula (I) according to claim 1, 2 or 3, or a pharmaceutically acceptable salt thereof, wherein the compound is selected from: 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-dihydro-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-dihydro-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-dihydro-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-dihydro-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-dihydro-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-dihydro-3H-1,2,4-triazol-3-one. 5. A method for preparing the compound of formula (I) according to any one of claims 1 to 4, characterized in that, firstly, the compound of formula (II) is prepared... It reacts with hydrazine to give the acylhydrazine of formula (III). Then, in the presence of a base, it condenses with amidine of formula (IV) or its salt. Wherein R ¹ has the meaning of any one of claims 1 to 4, This yields 1,2,4-triazole derivatives of formula (V) and/or their tautomers. Wherein R ¹ has the meaning of any one of claims 1 to 4, Then, in the presence of a copper catalyst and an amine base, it is coupled with phenylboronic acid of formula (VI). R ^2A and R ^2B have the meaning of any one of claims 1 to 4. The target compound of formula (I) was obtained. Wherein ^R1 , ^R2A , and ^R2B have the meaning of any one of claims 1 to 4. Then, optionally, under appropriate circumstances, the compounds of formula (I) are isolated to obtain their respective diastereomers, and/or the compounds of formula (I) are converted into their respective salts by treatment with the appropriate acid or base. 6. The compound of any one of claims 1 to 4, for the treatment and/or prevention of disease. 7. The compound of any one of claims 1 to 4, for the treatment and/or prevention of acute and chronic heart failure, cardiorenal syndrome, hypervolemic and normovolemic hyponatremia, cirrhosis, ascites, edema, and syndrome of inappropriate ADH secretion (SIADH). 8. Use of the compound of any one of claims 1 to 4 for the preparation of a pharmaceutical composition for the treatment and/or prevention of acute and chronic heart failure, cardiorenal syndrome, hypervolemic and normovolemic hyponatremia, cirrhosis, ascites, edema, and syndrome of inappropriate ADH secretion (SIADH). 9. A pharmaceutical composition comprising the compound of any one of claims 1 to 4 and one or more pharmaceutically acceptable excipients. 10. The pharmaceutical composition of claim 9 further comprises one or more additional therapeutic agents selected from diuretics, angiotensin II antagonists, ACE inhibitors, β-receptor blockers, mineralocorticoid receptor antagonists, NO donors, soluble guanylate cyclase stimulants, and positive inotropic agents. 11. The pharmaceutical composition of claim 9 further comprises an additional therapeutic agent selected from organic nitrates. 12. The pharmaceutical composition of claim 9 further comprises an additional therapeutic agent selected from soluble guanylate cyclase activators. 13. The pharmaceutical composition of any one of claims 9 to 12, for the treatment and/or prevention of acute and chronic heart failure, cardiorenal syndrome, hypervolemic and normovolemic hyponatremia, cirrhosis, ascites, edema, and syndrome of inappropriate ADH secretion (SIADH). 14. A compound 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 or a pharmaceutically acceptable salt thereof, said compound having the following formula: 15. The compound of claim 14, wherein the compound is 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, having the following formula: