Novel benzodiazepine derivatives as gaba a gamma1 pam

Abstract

To provide compounds for the treatment or prophylaxis of GABAAγ1 receptor-related diseases in a mammal, and diseases or conditions that can be treated by modulation of GABAAγ1 receptor activity.SOLUTION: There is provided a novel compound having the general formula (I), a composition comprising the compound, and a method using the compound.SELECTED DRAWING: None

Description

Technical Field

[0001] The present invention relates to organic compounds useful for the treatment or prevention in mammals, in particular, GABA A γ1 receptor-related diseases and diseases or conditions treatable by modulating GABA A γ1 receptor activity, such as autism spectrum disorder (ASD) targeting core symptoms and related co-existing conditions including anxiety and hypersensitivity, Angelman syndrome, Rett syndrome, Prader-Willi syndrome, fragile X disorder, psychosis, cognitive impairment and schizophrenia including negative symptoms, tardive dyskinesia, anxiety, separation anxiety disorder, selective mutism, specific phobia, social anxiety disorder (social phobia), panic disorder, agoraphobia, generalized anxiety disorder, substance / drug-induced anxiety disorder, impulse control and behavioral disorders, Tourette syndrome (TS), obsessive-compulsive disorder (OCD), acute stress disorder, post-traumatic stress disorder (PTSD), attention deficit hyperactivity disorder (ADHD), sleep disorders including narcolepsy attacks, Parkinson's disease (PD), neurodegenerative conditions including Huntington's disease, Alzheimer's disease (AD), mild cognitive impairment (MCI) dementia, behavioral and psychological symptoms in neurodegenerative conditions (BPS), multi-infarct dementia, psychosis and aggressive disorders, eating disorders including anorexia nervosa, bulimia nervosa, binge eating disorder, depression and related conditions (including treatment-resistant depression (TRD)), chronic apathy, anhedonia, chronic fatigue, seasonal affective disorder, postpartum depression, sexual dysfunction, bipolar disorder, epilepsy and pain for treating or preventing GABA A γ1 receptor positive allosteric modulators (PAMs).

Summary of the Invention

[0002] The present invention relates to formula (I)

Chemical formula

[0003] Receptors for the major inhibitory neurotransmitter gamma-aminobutyric acid (GABA) are mainly divided into two classes. (1) GABA A receptors, which are members of the ligand-gated ion channel superfamily, and (2) GABA B receptors, which are members of the G-protein coupled receptor family. The GABA A receptor complex, a membrane-bound heteropentameric protein polymer, is mainly composed of alpha, beta, and gamma subunits. The GABA A receptor is a ligand-bound chloride channel and a major mediator of inhibitory neurotransmission in the human brain.

[0004] GABA A There are 19 genes encoding the subunits of the receptor, and in the most common stoichiometry, two alpha, two beta, and one gamma subunit assemble as a pentamer. GABA ACombinations of subunits give rise to functional, circuit, and behavioral specificities (Sieghart, 2006; Vithlani et al., 2011). The GABA A receptor containing the γ1 subunit (GABA A γ1) is of particular interest because of their abundant expression in the limbic system (Seeburg et al., 1990; Pirker et al., 2000; Esmaeili et al., 2008; Durisic et al., 2017; Sequeira et al., 2019) as well as unique physiological and pharmacological properties (Mohler et al., 1996; Wingrove et al., 1997; Sieghart et al., 2005). GABA A receptors containing the γ1 subunit are not as numerous as those containing the γ2 subunit (about 5 - 10% of the total expression of GABA A receptors in the brain), but show high concentrations of brain mRNA and protein distribution in important brain regions such as the extended amygdala (central nucleus of the amygdala, medial nucleus, and bed nucleus), lateral septum, hypothalamus, and globus pallidus / substantia nigra. These structures form interconnected cores in subcortical limbic circuits that regulate stimulated social and emotional behaviors. In abnormal or disease states, overactivation of this circuit promotes anxiety, sexual arousal, aggression, fear, and defense while inhibiting feeding and social interactions (Goossens et al., 2007; Hofmann et al., 2011; Fox et al., 2012; Martin-Santos et al., 2014; Anderson et al., 2014; Calhoon et al., 2015).

[0005] Hyperactivity in the limbic cortical region, an important area for processing socially and emotionally relevant stimuli (known to form a functional network in coordination with the extended amygdala / hypothalamic region), is a common feature of various mental, neurological, neurodegenerative, neurodevelopmental, mood, motivational, and metabolic disorders. In such disease states, considering the characteristic anatomical distribution of GABA A receptors containing the γ1 subunit, GABAAγ1 positive allosteric modulators (PAMs) could be an effective treatment as symptomatic or disease-modifying agents.

[0006] Multiple lines of evidence suggest that an imbalance between excitatory / inhibitory (E / I) neurotransmission due to dysfunction of the GABAergic signaling system, a major inhibitory neurotransmission system in the brain, lies at the heart of the etiology of various CNS disorders. Considering the distribution and function of GABAA γ1 subunit-containing receptors in the CNS, they are very attractive targets for restoring the level of inhibition in important brain circuits and, as a result, for restoring the E / I balance in these conditions.

[0007] Accordingly, the compounds described herein, and their pharmaceutically acceptable salts and esters, alone or in combination with other drugs, are useful as disease-modifying or symptomatic agents for treating or preventing acute neuropathy, chronic neuropathy, cognitive impairment, autism spectrum disorder (ASD), Angelman syndrome, Rett syndrome, Prader-Willi syndrome, fragile X disorder, schizophrenia, tardive dyskinesia, anxiety, social anxiety disorder (social phobia), panic disorder, agoraphobia, generalized anxiety disorder, disruptive, impulse control and conduct disorders, Tourette syndrome (TS), obsessive-compulsive disorder (OCD), acute stress disorder, post-traumatic stress disorder (PTSD), attention deficit hyperactivity disorder (ADHD), sleep disorders, Parkinson's disease (PD), Huntington's disease, Alzheimer's disease (AD), mild cognitive impairment (MCI), dementia, behavioral and psychological symptoms in neurodegenerative states (BPS), multi-infarct dementia, agitation, psychosis, substance-induced psychotic disorder, aggression, eating disorders, depression, chronic apathy, anhedonia, chronic fatigue, seasonal affective disorder, postpartum depression, drowsiness, sexual dysfunction, bipolar disorder, epilepsy and pain.

[0008] The most preferred indications according to the present invention are anxiety disorders targeting social anxiety disorder (social phobia) and generalized anxiety disorder, and autism spectrum disorder (ASD) targeting core symptoms including anxiety and irritability and related co-existing disorders.

[0009] ASD is a complex and heterogeneous neurodevelopmental disorder characterized by two core domains: deficits in social communication and interaction, and the presence of repetitive or restricted behaviors, interests, or activities (American Psychiatric Association 2013).

[0010] There are no pharmacologic treatments approved for the core symptoms of social deficits and restricted / repetitive behaviors in ASD, and there are insufficient treatment options available for most of the emotional and physiological comorbidities of ASD. As a result, this disorder remains a highly unmet medical need. Currently approved treatments for ASD-related symptoms are limited to antipsychotics (risperidone and aripiprazole) that are indicated for the treatment of hypersensitivity associated with ASD symptoms. Emerging evidence suggests that the GABAergic system, a major inhibitory neurotransmitter system in the brain, plays an important role in the pathophysiology of ASD (Dhossche et al., 2002; Pizzarelli and Cherubini, 2011; Robertson et al., 2016).

[0011] Both genetic and imaging studies using positron emission tomography (PET) and magnetic resonance spectroscopy (MRS) have suggested changes in GABAergic signaling in ASD. The gene encoding GABAAγ1, GABRG1, is associated with α2, α4, and β1 GABA AIt is located on chromosome 4 (mouse Chr.5) within a cluster of genes encoding receptor subunits. A rare CNV containing an inversion of chromosome 4p12 that disrupts GABRG1 has been observed in siblings with autism (Horike et al., 2006) and similarly in the loss of GABRG1 in one case of ADHD. Mutations in the 4p12 gene cluster are associated with an increased risk of anxiety, substance abuse, and eating disorders, leading to an association between GABRG1 / 4p12 and mood dysfunction. MRS studies have found that GABA levels change in ASD (Gaetz et al., 2014; Rojas et al., 2014), and in particular, several recent studies have shown that GABA is decreased and somatosensory function is altered in children with ASD (Puts et al., 2016; Robertson et al., 2016). Consistent with these observations, a decrease in the number of inhibitory interneurons has been found in postmortem tissue from ASD and TS patients (Rapanelli et al., 2017). Furthermore, decreased levels of the GABA synthetic enzymes glutamate decarboxylase (GAD) 65 and 67 have been found in the frontal cortex and cerebellar cortex of autistic patients (Fatemi et al., 2002). Strong evidence in humans points to specific dysfunction in the limbic cortex region, which is known to form a functional network in coordination with the GABAAγ1 subunit-containing extended amygdala / hypothalamic region in ASD. These regions: cortex / lateral amygdala, insula, PFC, and lingual gyrus are recognized as key for processing socially and emotionally relevant stimuli. Subcortical nuclei that form specific partnerships with these regions to regulate behavioral outcomes are often difficult to study due to the limitations of spatial resolution, but much evidence points to overactivation of these cortico-subcortical connections in ASD. Furthermore, recent high-resolution studies have provided a clear link between increased amygdala activity / functional connectivity and emotional states (Kleinhans et al., 2009, 2016; Swartz et al., 2013; Nordahl et al., 2016; Ehrlich et al., 2017; Avino et al., 2018; Ibrahim et al., 2019).Targeting such very specific limbic subcortical regions that show substantial molecular and cellular diversity compared to the neocortex creates an accurate entry point for the safe and specific therapeutic modulation of the socio-emotional circuits affected in ASD while avoiding the broad regulation of the overall brain state. GABA by non-selective BZD. A Enhancement of receptor activity has been shown to improve behavioral impairments in mouse models of ASD, but GABA A A very narrow therapeutic margin was observed for sedation mediated by the α1γ2 subtype (Han et al., 2012, 2014; Soto et al., 2013). These findings support the idea that the balance of GABAergic transmission via the GABAAγ1 receptor may improve ASD symptoms without the side effects of non-selective benzodiazepines.

[0012] The object of the present invention is a compound of formula (I), and pharmaceutically acceptable salts and esters thereof, the preparation of the above compounds, pharmaceuticals containing them and their manufacture, and GABA A diseases associated with GABAAγ1 receptor dysfunction and GABA ADiseases or conditions treatable by enhancing γ1 receptor activity, such as autism spectrum disorder (ASD), Angelman syndrome, Rett syndrome, Prader-Willi syndrome, fragile X disorder, schizophrenia, tardive dyskinesia, anxiety, separation anxiety disorder, selective mutism, specific phobia, social anxiety disorder, panic disorder, agoraphobia, generalized anxiety disorder, substance / drug-induced anxiety disorder, disruptive, impulse control and conduct disorders, Tourette syndrome (TS), obsessive-compulsive disorder (OCD), acute stress disorder, post-traumatic stress disorder (PTSD), attention-deficit hyperactivity disorder (ADHD), sleep disorders including narcolepsy-cataplexy, Parkinson's disease (PD), Huntington's disease, Alzheimer's disease (AD), mild cognitive impairment (MCI), neurodegenerative conditions including dementia, behavioral and psychological symptoms in neurodegenerative conditions (BPS), multi-infarct dementia, psychosis and aggression, anorexia nervosa, bulimia nervosa, eating disorders including binge eating disorder, depression, and treatment-resistant depression (TRD), chronic apathy, anorexia, chronic fatigue, seasonal affective disorder, postpartum depression, drowsiness, sexual dysfunction, bipolar disorder, epilepsy and pain, and the use of the above compounds in the treatment or prevention of related conditions.

[0013] The compounds of the present invention increase the GABAergic current (influx of chloride) with a given concentration (e.g., EC 20 ) of gamma-aminobutyric acid (GABA) to selectively enhance the function of the γ1-containing GABA A receptor, and thus are selective GABA A γ1 receptor positive allosteric modulators (PAMs). The compounds of the present invention have a high PAM effect on γ1-containing subtypes (α5γ1, α2γ1, α1γ1) compared to γ2-containing subtypes (e.g., α1γ2, α2γ2, α3γ2, and α5γ2), and have high binding selectivity. Therefore, the compounds of the present invention are selective for γ2-containing GABA A subtypes and are strongly distinguished from classical benzodiazepine drugs such as alprazolam, triazolam, estazolam, midazolam, etc., which have low affinity for γ1-containing subtypes. Corresponding to the brain distribution of the γ1 subtype, selective GABA Aγ1 PAM will restore GABAergic signaling in key brain regions (e.g., the extended amygdala: terminal stria, lateral septum, hypothalamus, and central, medial, and bed nuclei of the pallidum / substantia nigra) without the side effects of non-selective GABA A modulators (e.g., benzodiazepines).

[0014] The term "amino" represents the -NH2 group.

[0015] "Amino-C" 1-6 -alkyl" refers to a C 1-6 -alkyl group in which one of the hydrogen atoms of the C 1-6 -alkyl group is replaced by an amino group. Examples of amino-C 1-6 -alkyl include aminomethyl, aminoethyl, aminopropyl, aminomethylpropyl, aminomethylethyl, and aminobutyl. A specific example is aminomethyl.

[0016] "C 1-6 -alkoxy" refers to a group of the formula -O-R', where R' represents a C 1-6 -alkyl group. Examples of C 1-6 -alkoxy groups include methoxy, ethoxy, n-propoxy, isopropoxy, n-butoxy, isobutoxy, and tert-butoxy. Specific examples are methoxy and ethoxy. A more specific example is methoxy.

[0017] "C 1-6 -alkoxy-C 1-6 -alkyl" refers to a C 1-6 -alkyl group in which at least one of the hydrogen atoms of the C 1-6 -alkyl group is replaced by a C 1-6 -alkoxy group. Exemplary C 1-6 -alkoxy-C 1-6 -alkyl groups include methoxymethyl, ethoxymethyl, methoxymethyl, ethoxyethyl, methoxypropyl, ethoxypropyl, etc. A specific example is methoxymethyl.

[0018] "C 1-6 -alkyl" refers to a monovalent straight-chain or branched-chain saturated hydrocarbon group having 1 to 6 carbon atoms. C 1-6 Examples of C 1-6 -alkyl include methyl, ethyl, n-propyl, isopropyl, n-butyl, iso-butyl, sec-butyl, tert-butyl, and pentyl. Specific C

[0019] "C 3~8 -cycloalkyl" refers to a monovalent saturated monocyclic or bicyclic hydrocarbon group having 3 to 8 ring carbon atoms. Bicyclic means a ring system consisting of 2 saturated carbon rings having 1 or 2 carbon atoms in common. Monocyclic C 3-8 -cycloalkyl examples are cyclopropyl, cyclobutanil, cyclopentyl, cyclohexyl or cycloheptyl. Bicyclic C 3-8 -cycloalkyl example is spiro[3.3]heptanyl. Specific monocyclic C 3-8 -cycloalkyl groups are cyclopropyl and cyclobutanil. A more specific monocyclic C 3-8 -cycloalkyl group includes cyclopropyl.

[0020] The term "cyano" represents a -CN group.

[0021] "Halo-C 1-6 -alkyl" refers to a C 1-6 -alkyl group in which at least one hydrogen atom of the C 1-6 -alkyl group is substituted with the same or different halogen atoms. "Perhalo-C 1-6 -alkyl-C 1-6 -alkyl" refers to a C 1-6 -alkyl-C 1-6 -alkyl group in which all hydrogen atoms of the alkyl group are substituted with the same or different halogen atoms. Halo-C 1-6Examples of -alkyl include fluoromethyl, difluoromethyl, trifluoromethyl, fluoroethyl, difluoroethyl, and trifluoroethyl. More specific halo-C 1-6 Examples of -alkyl groups include trifluoromethyl and difluoromethyl. More specific halo-C 1-6 The -alkyl group is trifluoromethyl.

[0022] The terms "halogen" and "halo" are used interchangeably herein and represent fluoro, chloro, bromo, or iodo. Specific halogens include fluoro, chloro, etc.

[0023] The term "heteroaryl" refers to a monovalent aromatic heterocyclic or monocyclic or bicyclic ring system consisting of 5 to 12 ring atoms composed of 1, 2, 3, or 4 heteroatoms selected from N, O, and S, with the remaining ring atoms being carbon. Examples of heteroaryl include pyrrolyl, furanyl, thienyl, imidazolyl, oxazolyl, thiazolyl, triazolyl, oxadiazolyl, thiadiazolyl, tetrazolyl, pyridinyl, pyrazinyl, pyrazolyl, pyridazinyl, pyrimidinyl, triazinyl, azepinyl, diazepinyl, isoxazolyl, benzofuranyl, isothiazolyl, benzothienyl, indolyl, isoindolyl, isobenzofuranyl, benzimidazolyl, benzoxazolyl, benzoisoxazolyl, benzoisothiazolyl, benzoxadiazolyl, benzothiazolyl, benzotriazolyl, purinyl, quinolinyl, isoquinolinyl, quinazolinyl, quinoxalinyl. Specific heteroaryl groups include pyridinyl, pyrazolyl, pyrimidinyl, pyridazinyl, and isoxazolyl. More specific heteroaryl groups are pyrazolyl, pyrimidinyl, pyridazinyl, and isoxazolyl.

[0024] The term "heterocycloalkyl" refers to a monocyclic or bicyclic ring system consisting of 4 to 11 ring atoms, including 1, 2, or 3 ring heteroatoms selected from N, O, and S, with the remaining ring atoms being carbon. Bicyclic means consisting of two rings that share one or two ring atoms in common. Examples of monocyclic saturated heterocyclyl include 4,5-dihydro-oxazolyl, oxetanyl, azetidinyl, pyrrolidinyl, 2-oxo-pyrrolidin-3-yl, tetrahydrofuranyl, tetrahydrothiophenyl, pyrazolidinyl, imidazolidinyl, oxazolidinyl, isoxazolidinyl, thiazolidinyl, piperidinyl, tetrahydropyranyl, tetrahydrothiopyranyl, piperazinyl, morpholinyl, thiomorpholinyl, 1,1-dioxo-thiomorpholin-4-yl, azepanyl, diazepanyl, homopiperazinyl, or oxazepanyl. Examples of bicyclic saturated heterocycloalkyl include oxabicyclo[2.2.1]heptanyl, oxaspiro[3.3]heptanyl, 8-aza-bicyclo[3.2.1]octyl, quinuclidinyl, 8-oxa-3-aza-bicyclo[3.2.1]octyl, 9-aza-bicyclo[3.3.1]nonyl, 3-oxa-9-aza-bicyclo[3.3.1]nonyl, or 3-thia-9-aza-bicyclo[3.3.1]nonyl. Examples of partially unsaturated heterocycloalkyl include dihydrofuryl, imidazolinyl, dihydrooxazolyl, tetrahydropyridinyl, or dihydropyranyl. A particular heterocycloalkyl is tetrahydropyranyl.

[0025] The term "hydroxy" represents an -OH group.

[0026] "Hydroxy-C 1-6 -alkyl" refers to a C 1-6 -alkyl group in which one of the hydrogen atoms of the C 1-6 -alkyl group is replaced by a hydroxy group. Hydroxy C 1-6Examples of -alkyl include hydroxymethyl, hydroxyethyl, hydroxypropyl, hydroxymethylpropyl, hydroxymethylethyl, hydroxybutyl, etc. A specific example is hydroxymethyl.

[0027] The term "pharmaceutically acceptable salt" refers to salts that retain the biological effectiveness and properties of the free base or free acid and are not biologically or otherwise undesirable. The salts are formed with inorganic acids such as hydrochloric acid, hydrobromic acid, sulfuric acid, nitric acid, phosphoric acid, especially hydrochloric acid, and organic acids such as acetic acid, propionic acid, glycolic acid, pyruvic acid, oxalic acid, maleic acid, malonic acid, succinic acid, fumaric acid, tartaric acid, citric acid, benzoic acid, cinnamic acid, mandelic acid, methanesulfonic acid, ethanesulfonic acid, p-toluenesulfonic acid, salicylic acid, N-acetylcysteine, etc. Also, these salts may be prepared by adding an inorganic base or an organic base to the free acid. Salts derived from inorganic bases include, but are not limited to, sodium, potassium, lithium, ammonium, calcium, and magnesium salts. Salts derived from organic bases include primary, secondary, and tertiary amines, substituted amines including naturally occurring substituted amines, cyclic amines, and basic ion exchange resins, such as salts of isopropylamine, trimethylamine, diethylamine, triethylamine, tripropylamine, ethanolamine, lysine, arginine, N-ethylpiperidine, piperidine, polyimine resins, etc., but are not limited thereto. Particularly pharmaceutically acceptable salts of the compound of formula (I) are hydrochloride, methanesulfonate, and citrate.

[0028] "Pharmaceutically acceptable ester" means that a compound of general formula (I) can be derivatized with a functional group to provide a derivative that can be converted back to the parent compound in vivo. Examples of such compounds include physiologically acceptable and metabolically unstable ester derivatives such as methoxymethyl ester, methylthiomethyl ester, and pivaloyloxymethyl ester. Furthermore, any physiologically acceptable equivalents of the compounds of general formula (I) that are similar to metabolically unstable esters that can produce the parent compound of general formula (I) in vivo are also within the scope of the present invention.

[0029] The term "protecting group" (PG) means a group that selectively blocks the reactive sites of a polyfunctional compound and enables selective chemical reactions to be carried out at other unprotected reactive sites, and is used in its conventional meaning in synthetic chemistry. The protecting group can be removed at an appropriate point. Exemplary protecting groups are amino protecting groups, carboxy protecting groups, or hydroxy protecting groups. Specific protecting groups are the tert-butoxycarbonyl (Boc) group, benzyloxycarbonyl (Cbz) group, fluorenylmethoxycarbonyl (Fmoc) group, and benzyl (Bn) group. More specific protecting groups are the tert-butoxycarbonyl (Boc) group and fluorenylmethoxycarbonyl (Fmoc) group. Even more specific protecting group is the tert-butoxycarbonyl (Boc) group.

[0030] The abbreviation uM means micromole and corresponds to the symbol μM.

[0031] The abbreviation uL means microliter and corresponds to the symbol μL.

[0032] The abbreviation ug means microgram and corresponds to the symbol μg.

[0033] The compounds of formula (I) can contain several asymmetric centers and can exist in the form of optically pure enantiomers, mixtures of enantiomers such as racemates, optically pure diastereoisomers, mixtures of diastereoisomers, racemates of diastereoisomers or mixtures of racemates of diastereoisomers.

[0034] According to the Cahn-Ingold-Prelog rules, the asymmetric carbon atoms can have the "R" or "S" configuration.

[0035] Also, embodiments of the present invention are compounds according to formula (I) described herein and pharmaceutically acceptable salts or esters thereof, in particular compounds according to formula (I) described herein and pharmaceutically acceptable salts thereof, more specifically, compounds according to formula (I) described herein.

[0036] Specific embodiments of the present invention are compounds according to formula (I) described herein, wherein R 1 is i) H, ii) C 1-6 -alkyl, iii) C 1-6 -alkoxy, iv) C 1-6 -alkoxy-C 1-6 -alkyl, v) hydroxy, vi) hydroxy-C 1-6 -alkyl, vii) optionally R 7 , R 8 and R 9 substituted C 3-8 -cycloalkyl, viii) amino-C 1-6 -alkyl, ix) optionally R 7 , R 8 and R 9 substituted pyrazolyl, x) optionally R 7 , R 8 and R 9 substituted pyridinyl, xi) optionally substituted by R 7 , R 8 and R 9 substituted pyrimidinyl, xii) optionally substituted by R 7 , R 8 and R 9 substituted pyridazinyl, xiii) optionally substituted by R 7 , R 8 and R 9 substituted isoxazolyl selected from; R 2 is i) C 1-6 -alkyl, ii) hydroxy, iii) hydroxy-C 1-6 -alkyl, and iv) C 1-6 -alkoxy-C 1-6 -alkyl selected from; R 3 is i) Cl, and ii) F selected from, X is i) CR 6 , and ii) N selected from; R 6 is i) H, ii) Cl, and iii) F selected from; R 4 is i) Br, and ii) Cl selected from; R 5 is i) C 1-6 -alkyl, ii) C 1-6 -alkoxy, iii) halogen, iv) halo-C 1-6 -alkyl, v)C 3-8 -cycloalkyl selected from; R 7 、R 8 and R 9 are, i)C 1-6 -alkyl, and ii)C 1-6 -alkoxy selected from) or a pharmaceutically acceptable salt thereof is provided.

[0037] A more specific embodiment of the present invention is a compound of formula (I) according to claim 1, wherein R 1 is, i) H, ii) C 1-6 -alkyl, iii) hydroxy, iv) hydroxy-C 1-6 -alkyl, v) optionally R 7 、R 8 and R 9 substituted C 3-8 -cycloalkyl, vi) optionally R 7 、R 8 and R 9 substituted pyrazolyl, vii) optionally R 7 、R 8 and R 9 substituted pyrimidinyl, viii) optionally R 7 、R 8 and R 9 substituted pyridazinyl, ix) optionally R 7 、R 8 and R 9 substituted isoxazolyl selected from; R 2 is, i) C 1-6 -alkyl, ii) hydroxy, iii) hydroxy-C 1-6 -alkyl, and iv) C 1-6 -alkoxy-C 1-6 -alkyl is selected from; R 3 is F, X is i) CR 6 and ii) N is selected from; R 6 is i) H, and ii) F is selected from; R 4 is i) Br, and ii) Cl is selected from; R 5 is i) C 1-6 -alkyl, ii) halogen, and iii) halo-C 1-6 -alkyl is selected from; R 7 、R 8 and R 9 are independently selected from C 1-6 alkyl, to provide a compound or a pharmaceutically acceptable salt thereof.

[0038] A further specific embodiment of the present invention is a compound according to formula (I) described herein, R 1 is C 1-6 alkyl, R 2 is C 1-6 alkyl, R 3 is F, X is CR 6 and R 6 is F, R 4 is Cl, R 5 is halo-C 1-6 -alkyl, a compound, or a pharmaceutically acceptable salt thereof.

[0039] Another specific embodiment of the present invention is that R 1 is i) H, ii) C 1-6 -alkyl, iii) C 1-6 -alkoxy, iv) C 1-6 -alkoxy-C 1-6 -alkyl, v) hydroxy, vi) hydroxy-C 1-6 -alkyl, vii) optionally R7, R 8 and R 9 substituted C 3-8 cycloalkyl, viii) amino-C 1-6 -alkyl, ix) optionally R 7 , R 8 and R 9 substituted pyrazolyl, x) optionally R 7 , R 8 and R 9 substituted pyridinyl, xi) optionally R 7 , R 8 and R 9 substituted pyrimidinyl, xii) optionally R 7 , R 8 and R 9 substituted pyridazinyl, and xiii) optionally R 7 , R 8 and R 9 substituted isoxazolyl selected from the compounds of formula (I) described herein.

[0040] Another specific embodiment of the present invention is that R 1is i) H, ii) C 1-6 -alkyl, iii) hydroxy, iv) hydroxy-C 1-6 -alkyl, v) optionally R7, R 8 and R 9 substituted C 3-8 -cycloalkyl, vi) optionally R 7 , R 8 and R 9 substituted pyrazolyl, vii) optionally R 7 , R 8 and R 9 substituted pyrimidinyl, viii) optionally R 7 , R 8 and R 9 substituted pyridazinyl, ix) optionally R 7 , R 8 and R 9 substituted isoxazolyl selected from the compounds of formula (I) described herein are provided.

[0041] Another specific embodiment of the present invention provides a compound of formula (I) described herein, wherein R 1 is C 1-6 -alkyl.

[0042] A specific embodiment of the present invention provides a compound of formula (I) described herein, wherein R 2 is C 1-6 alkyl.

[0043] Another specific embodiment of the present invention provides a compound of formula (I) described herein, wherein R 3 is F.

[0044] Another specific embodiment of the present invention provides a compound according to formula (I) described herein, wherein X is CR 6 .

[0045] Another specific embodiment of the present invention provides a compound of formula (I) as described herein, wherein R 6 is F.

[0046] Another specific embodiment of the present invention provides a compound of formula (I) as described herein, wherein R 4 is Cl.

[0047] Another specific embodiment of the present invention provides a compound of formula (I) as described herein, wherein R 5 is selected from i) C 1-6 -alkyl, ii) C 1-6 -alkoxy, iii) halogen, iv) halo-C 1-6 -alkyl, v) C 3-8 -cycloalkyl Another specific embodiment of the present invention provides a compound of formula (I) as described herein, wherein R

[0048] Another specific embodiment of the present invention provides a compound of formula (I) as described herein, wherein R 5 is selected from i) C 1-6 -alkyl, ii) halogen, and iii) halo-C 1-6 -alkyl Another specific embodiment of the present invention provides a compound of formula (I) as described herein, wherein R

[0049] Another specific embodiment of the present invention provides a compound of formula (I) as described herein, wherein R 5 is C 1-6 alkyl.

[0050] Another specific embodiment of the present invention provides a compound of formula (I) as described herein, wherein R 7 , R 8 and R 9 are independently selected from C 1-6 -alkyl.

[0051] Specific examples of the compounds of formula (I) described herein are (4S)-8-Bromo-7-chloro-6-(2-fluorophenyl)-4-methyl-4H-[1,2,4]triazolo[4,3-a][1,4]benzodiazepine; 8-Bromo-7-chloro-6-(2,6-difluorophenyl)-4H-[1,2,4]triazolo[4,3-a][1,4]benzodiazepin-4-ol; (4S)-8-Bromo-7-chloro-6-(2-fluorophenyl)-4-methyl-1-pyridazin-3-yl-4H-[1,2,4]triazolo[4,3-a][1,4]benzodiazepine; 8-Bromo-7-chloro-6-(2-fluorophenyl)-1-pyridazin-3-yl-4H-[1,2,4]triazolo[4,3-a][1,4]benzodiazepin-4-ol; (4S)-8-Bromo-7-chloro-6-(2,6-difluorophenyl)-4-methyl-4H-[1,2,4]triazolo[4,3-a][1,4]benzodiazepine; (4S)-7,8-Dichloro-6-(2,6-difluorophenyl)-4-methyl-1-pyrimidin-4-yl-4H-[1,2,4]triazolo[4,3-a][1,4]benzodiazepine; (4S)-7,8-Dichloro-6-(2,6-difluorophenyl)-4-methyl-1-(1-methylpyrazol-4-yl)-4H-[1,2,4]triazolo[4,3-a][1,4]benzodiazepine; (4S)-7,8-Dichloro-6-(2,6-difluorophenyl)-4-methyl-1-pyridazin-3-yl-4H-[1,2,4]triazolo[4,3-a][1,4]benzodiazepine; (4S)-7,8-Dichloro-6-(3-fluoro-2-pyridyl)-4-methyl-1-pyridazin-3-yl-4H-[1,2,4]triazolo[4,3-a][1,4]benzodiazepine; (4S)-7,8-Dichloro-6-(2,6-difluorophenyl)-4-methyl-2,4-dihydro-[1,2,4]triazolo[4,3-a][1,4]benzodiazepin-1-one; (4S)-7,8-Dichloro-6-(3-fluoro-2-pyridyl)-4-methyl-1-pyrimidin-4-yl-4H-[1,2,4]triazolo[4,3-a][1,4]benzodiazepine; (4S)-7,8-Dichloro-6-(3-fluoro-2-pyridyl)-4-methyl-1-(1-methylpyrazol-4-yl)-4H-[1,2,4]triazolo[4,3-a][1,4]benzodiazepine; (4S)-7,8-Dichloro-1-cyclopropyl-6-(3-fluoro-2-pyridyl)-4-methyl-4H-[1,2,4]triazolo[4,3-a][1,4]benzodiazepine; (4S)-7-Bromo-8-chloro-6-(2,6-difluorophenyl)-4-methyl-1-pyridazin-3-yl-4H-[1,2,4]triazolo[4,3-a][1,4]benzodiazepine; (4S)-7-Bromo-8-chloro-6-(2,6-difluorophenyl)-4-methyl-1-pyrimidin-4-yl-4H-[1,2,4]triazolo[4,3-a][1,4]benzodiazepine; (4S)-8-Bromo-7-chloro-6-(2,6-difluorophenyl)-1,4-dimethyl-4H-[1,2,4]triazolo[4,3-a][1,4]benzodiazepine; (4S)-7-Bromo-8-chloro-6-(2,6-difluorophenyl)-4-methyl-1-(1-methylpyrazol-4-yl)-4H-[1,2,4]triazolo[4,3-a][1,4]benzodiazepine; (4S)-7-Bromo-8-chloro-6-(2,6-difluorophenyl)-1,4-dimethyl-4H-[1,2,4]triazolo[4,3-a][1,4]benzodiazepine; (4S)-7-Bromo-8-chloro-6-(2,6-difluorophenyl)-4-methyl-1-(1-methylpyrazol-3-yl)-4H-[1,2,4]triazolo[4,3-a][1,4]benzodiazepine; (4S)-7,8-Dichloro-6-(2,6-difluorophenyl)-1,4-dimethyl-4H-[1,2,4]triazolo[4,3-a][1,4]benzodiazepine; (4S)-8-Bromo-7-chloro-1-cyclopropyl-6-(3-fluoro-2-pyridyl)-4-methyl-4H-[1,2,4]triazolo[4,3-a][1,4]benzodiazepine; (4S)-8-Bromo-7-chloro-6-(3-fluoro-2-pyridyl)-4-methyl-4H-[1,2,4]triazolo[4,3-a][1,4]benzodiazepine; (4S)-8-Bromo-7-chloro-6-(3-fluoro-2-pyridyl)-1,4-dimethyl-4H-[1,2,4]triazolo[4,3-a][1,4]benzodiazepine; (4S)-7,8-Dichloro-6-(2,6-difluorophenyl)-4-methyl-1-(6-methylpyridazin-3-yl)-4H-[1,2,4]triazolo[4,3-a][1,4]benzodiazepine; 5-[(4S)-7,8-Dichloro-6-(2,6-difluorophenyl)-4-methyl-4H-[1,2,4]triazolo[4,3-a][1,4]benzodiazepin-1-yl]-3-methyl-isoxazole; (4S)-7-Bromo-8-chloro-6-(2,6-difluorophenyl)-4-methyl-1-(6-methylpyridazin-3-yl)-4H-[1,2,4]triazolo[4,3-a][1,4]benzodiazepine; (4S)-7-Bromo-8-chloro-6-(2,6-difluorophenyl)-4-methyl-2,4-dihydro-[1,2,4]triazolo[4,3-a][1,4]benzodiazepin-1-one; (4S)-7-Chloro-6-(2,6-difluorophenyl)-1,4-dimethyl-8-(trifluoromethyl)-4H-[1,2,4]triazolo[4,3-a][1,4]benzodiazepine; (4S)-7-Chloro-8-(1,1-difluoroethyl)-6-(2,6-difluorophenyl)-1,4-dimethyl-4H-[1,2,4]triazolo[4,3-a][1,4]benzodiazepine; (4S)-7,8-Dichloro-6-(2,6-difluorophenyl)-4-methyl-1-(6-methylpyrimidin-4-yl)-4H-[1,2,4]triazolo[4,3-a][1,4]benzodiazepine; (4S)-7,8-Dichloro-6-(2,6-difluorophenyl)-4-methyl-1-(2-methylpyrimidin-4-yl)-4H-[1,2,4]triazolo[4,3-a][1,4]benzodiazepine; (4S)-7,8-Dichloro-6-(2,6-difluorophenyl)-1-(2,6-dimethylpyrimidin-4-yl)-4-methyl-4H-[1,2,4]triazolo[4,3-a][1,4]benzodiazepine; (4S)-7-Chloro-6-(2,6-difluorophenyl)-1,4,8-trimethyl-4H-[1,2,4]triazolo[4,3-a][1,4]benzodiazepine; (4S)-8-Bromo-7-chloro-6-(2,6-difluorophenyl)-4-ethyl-1-methyl-4H-[1,2,4]triazolo[4,3-a][1,4]benzodiazepine; (4S)-7-Chloro-8-(difluoromethyl)-6-(2,6-difluorophenyl)-1,4-dimethyl-4H-[1,2,4]triazolo[4,3-a][1,4]benzodiazepine; (4R)-8-Bromo-7-chloro-6-(2,6-difluorophenyl)-4-(methoxymethyl)-1-methyl-4H-[1,2,4]triazolo[4,3-a][1,4]benzodiazepine; [(4R)-8-Bromo-7-chloro-6-(2,6-difluorophenyl)-1-methyl-4H-[1,2,4]triazolo[4,3-a][1,4]benzodiazepin-4-yl]methanol; (4S)-7-Chloro-6-(2,6-difluorophenyl)-8-iodo-1,4-dimethyl-4H-[1,2,4]triazolo[4,3-a][1,4]benzodiazepine; [(4S)-7-chloro-6-(2,6-difluorophenyl)-4-methyl-8-(trifluoromethyl)-4H-[1,2,4]triazolo[4,3-a][1,4]benzodiazepin-1-yl]methanol; (4R)-7-chloro-6-(2,6-difluorophenyl)-4-(methoxymethyl)-1-methyl-8-(trifluoromethyl)-4H-[1,2,4]triazolo[4,3-a][1,4]benzodiazepine; [(4R)-7-chloro-6-(2,6-difluorophenyl)-1-methyl-8-(trifluoromethyl)-4H-[1,2,4]triazolo[4,3-a][1,4]benzodiazepin-4-yl]methanol; (4S)-7-chloro-6-(2,6-difluorophenyl)-4-methyl-8-(trifluoromethyl)-4H-[1,2,4]triazolo[4,3-a][1,4]benzodiazepine; (4S)-7-chloro-6-(2,6-difluorophenyl)-1-ethyl-4-methyl-8-(trifluoromethyl)-4H-[1,2,4]triazolo[4,3-a][1,4]benzodiazepine; (4S)-7-chloro-6-(2,6-difluorophenyl)-8-ethyl-1,4-dimethyl-4H-[1,2,4]triazolo[4,3-a][1,4]benzodiazepine; or a pharmaceutically acceptable salt thereof selected from.

[0052] Further specific examples of the compounds of formula (I) described herein are selected from (4S)-7-chloro-6-(2,6-difluorophenyl)-1,4-dimethyl-8-(trifluoromethyl)-4H-[1,2,4]triazolo[4,3-a][1,4]benzodiazepine, or a pharmaceutically acceptable salt thereof.

[0053] In some embodiments, the compounds of formula (I) are isotopically labeled by replacing one or more atoms therein with atoms having a different atomic mass or mass number. Such isotopically labeled (i.e., radiolabeled) compounds of formula (I) are considered to be within the scope of the present disclosure. Examples of isotopes that can be incorporated into the compounds of formula (I) include isotopes of hydrogen, carbon, nitrogen, oxygen, phosphorus, sulfur, fluorine, chlorine, and iodine, for example, but not limited to, 2 H, 3 H, 11 C, 13 C, 14 C, 13 N, 15 N, 15 O, 17 O, 18 O, 31 P, 32 P, 35 S, 18 F, 36 Cl, 123 I, and 125 I. Some isotopically labeled compounds of formula (I), for example, those incorporating radioisotopes, are useful in drug and / or substrate tissue distribution studies. The radioisotopes tritium, i.e., 3 H, and carbon-14, i.e., 14 C are particularly useful for this purpose in view of their ease of incorporation and ready means of detection. For example, the compounds of formula (I) can be enriched with 1, 2, 5, 10, 25, 50, 75, 90, 95, or 99 percent of a given isotope.

[0054] Replacement with heavier isotopes, such as deuterium, i.e., 2 H, etc., can result in higher metabolic stability, for example, by increasing the in vivo half-life or decreasing the required dosage, thereby providing certain therapeutic advantages.

[0055] 1 1 C, 18 F, 15 O and 13Substitution with positrons that emit isotopes such as N can be useful in positron emission tomography (PET) for examining substrate receptor occupancy. Isotope-labeled compounds of formula (I) can generally be prepared by methods similar to those described in the examples below, which use appropriate isotope-labeled reagents in place of the previously used unlabeled reagents, or by conventional techniques known to those skilled in the art.

[0056] Also an object of the present invention is a method for producing the compounds of formula (I) described herein.

[0057] The preparation of the compounds of formula (I) of the present invention can be carried out by sequential or convergent synthetic routes. The synthesis of the present invention is shown in the following general schemes. The skills necessary to carry out the reactions and the purification of the resulting products are known to those skilled in the art. The substituents and indicators used in the following description of the process have the significance described above herein unless otherwise noted.

[0058] More specifically, the compounds of formula (I) can be prepared by the methods shown below, the methods shown in the examples, or similar methods. Appropriate reaction conditions for the individual reaction steps are known to those skilled in the art. The order of the reactions is not limited to that shown in Schemes 1-4, but the order of the reaction steps can be freely changed according to the starting materials and their respective reactivities. The starting materials can be commercially available or can be prepared by methods similar to those shown below, the methods described in the references or examples cited herein, or methods known in the art.

[0059] The present compounds of formula (I) and their pharmaceutically acceptable salts can be prepared by the methods described below (Scheme 1).

Chemical formula

[0060] Scheme 1: Synthesis of benzodiazepine (I) where all definitions are as described above and in the claims According to Scheme 1, the compound of formula (I) can be prepared in two steps starting from the lactam of formula (II) (constituents A, B, G, L, M, O, Q-U). After the vulcanization reaction using Lawesson's reagent or P2S5, the lactam (II) is converted to the corresponding thiolactam (III). Their reaction with hydrazide (IV) via a Perizzari-type process gives the 1,2,4-triazole of general formula (I). Alternatively, 1,2,4-triazole (I) can be obtained by reacting thiolactam (II) with hydrazine to form hydrazone (V), followed by treatment with triethyl orthoacetate or triethyl orthoformate.

[0061] R 1 In a particular embodiment of the invention where R is hydroxyl (OH), the benzodiazepine of formula (I) can be obtained in two steps according to the process described in Scheme 2. It is widely accepted that 3-hydroxy-1,2,4-triazole exists as two tautomeric forms, and in the present invention, they are exclusively represented in their most stable form (triazolone). For this purpose, hydrazone (V) can be reacted with 1,1'-carbonyldiimidazole (CDI) to obtain the triazolone of formula (I) (Scheme 2).

Chemical formula

[0062] Scheme 2: R 1 Synthesis of benzodiazepine (I) where R is hydroxyl; all other definitions are as described above and in the claims. The synthesis of the components (A, B, G, L, M, O, Q - U) of formula (II) is highlighted in Scheme 3. Commercially available 2 - amino - 6 - chlorobenzoic acid or 2 - amino - 6 - bromobenzoic acid can be heated in acetic anhydride to form 5 - chloro - 2 - methyl - 3,1 - benzoxazin - 4 - one and 5 - bromo - 2 - methyl - 3,1 - benzoxazin - 4 - one, respectively. A Grignard reagent or an organolithium reagent of formula (VI) (prepared by metallation reaction or kinetic deprotonation from the corresponding aryl bromide) is reacted with benzoxazin - 4 - one (electrophile) at a controlled temperature to obtain a ketone of formula (VII). After N - acetamido hydrolysis under acidic conditions (HCl), the compound of formula (VII) is converted to an aniline of formula (VIII). Conveniently, at this juncture, the halogen of R 5 can be introduced by treatment with N - chlorosuccinimide (NCS), N - bromosuccinimide (NBS) or N - iodosuccinimide (NIS) to obtain an intermediate of formula (IX). The final thermal cyclization reaction with ethyl 2 - aminoacetate hydrochloride in pyridine gives the desired benzodiazepine (II), presumably via the formation of an imine intermediate (X). [Chemical formula]

[0063] Scheme 3: Synthesis of the components (A, B, G, L, M, O, Q - U) where R 5 is Cl, Br or I and R 2 is H; all other definitions are as described above and in the claims. R 2 In a further embodiment of the invention where R is alkyl or substituted alkyl, an alternative process is envisioned and detailed in Scheme 4. [Chemical formula]

[0064] Scheme 4: R 5 is Cl, Br or I and R 2Synthesis of components (A, B, G, L, M, O, Q-U) that are alkyl or substituted alkyl; all other definitions are as described above and in the claims. In such cases, the compound of formula (XI) can be prepared by an amide coupling reaction between aniline (IX) and an N-Boc protected L-amino acid in the presence of phosphoryl chloride (POCl3), or by other methods known to those skilled in the art. Removal of the N-Boc protecting group can be carried out using a mineral acid (e.g., HCl) or an organic acid (e.g., trifluoroacetic acid) to obtain the amine of formula (XII). By promoting the final intramolecular condensation reaction with an acidic medium (e.g., silica or acetic acid) and heat (80 - 110 °C), the desired benzodiazepine components (A - U) of formula (II) are obtained. In particular, in the processes described in Schemes 1 and 4, depending on the specific reaction conditions employed, the degree of racemization at the chiral center varies (20 - 100%). As a result, chiral purification of the final derivative of formula (I) (e.g., by HPLC or SFC) is necessary to obtain a final derivative with an enantiomeric excess (ee) exceeding 97%.

[0065] Also, an embodiment of the present invention is a process for preparing a compound of formula (I) as defined above, which includes the reaction of a compound of formula (III) and a compound of formula (IV) in a solvent, particularly an alcohol such as butan-1-ol, at a temperature between room temperature and the reflux temperature of the solvent, particularly at the reflux temperature of the solvent.

Chemical formula

[0066] Also, an object of the present invention is a compound of formula (I) for use as a therapeutic active substance, more specifically a compound of formula (I) as described herein.

[0067] Similarly, an object of the present invention is a pharmaceutical composition comprising a compound of formula (I), more particularly a compound of formula (I) described herein and a therapeutically inert carrier.

[0068] Certain embodiments of the invention are for use in the treatment or prevention of Alzheimer's disease, mild cognitive impairment (MCI), age-related cognitive decline, negative and / or cognitive symptoms associated with schizophrenia, bipolar disorder, autism spectrum disorder (ASD), Angelman syndrome, Rett syndrome, Prader-Willi syndrome, epilepsy, post-traumatic stress disorder (PTSD), amyotrophic lateral sclerosis (ALS), fragile X disorder, more particularly negative and / or cognitive symptoms associated with autism spectrum disorder (ASD), Angelman syndrome, Alzheimer's disease, schizophrenia and post-traumatic stress disorder (PTSD), more particularly for use in treatment, a compound of formula (I) described herein or a pharmaceutically acceptable salt thereof.

[0069] The present invention also relates to a compound of formula (I) described herein or a pharmaceutically acceptable salt thereof, more particularly for use in the preparation of a medicament for treating or preventing, more particularly treating, negative symptoms and / or cognitive symptoms associated with Alzheimer's disease, mild cognitive impairment (MCI), age-related cognitive decline, negative symptoms and / or cognitive symptoms associated with schizophrenia, bipolar disorder, autism spectrum disorder (MCIASD), Angelman syndrome, Rett syndrome, Prader-Willi syndrome, epilepsy, post-traumatic stress disorder (PTSD), amyotrophic lateral sclerosis (ALS), fragile X disorder, more particularly negative symptoms and / or cognitive symptoms associated with autism spectrum disorder (ASD), Angelman syndrome, Alzheimer's disease, schizophrenia and post-traumatic stress disorder (PTSD).

[0070] Furthermore, an object of the present invention is to provide a method for treating or preventing negative symptoms and / or cognitive symptoms associated with Alzheimer's disease, mild cognitive impairment (MCI), age-related decline in cognitive function, schizophrenia, bipolar disorder, autism spectrum disorder (ASD), Angelman syndrome, Rett syndrome, Prader-Willi syndrome, epilepsy, post-traumatic stress disorder (PTSD), amyotrophic lateral sclerosis (ALS), fragile X disorder, particularly negative symptoms and / or cognitive symptoms associated with autism spectrum disorder (ASD), Angelman syndrome, Alzheimer's disease, schizophrenia, and post-traumatic stress disorder (PTSD), more particularly a method for treatment, which comprises administering an effective amount of a compound of formula (I) described herein, more specifically a compound of formula (I).

[0071] Moreover, one embodiment of the present invention is a compound of formula (I), more specifically a compound of formula (I) as described herein, when manufactured according to any one of the processes described.

[0072] Assay procedure GABA containing γ1 A Membrane preparation and binding assay for subtypes GABA A The affinity of a compound for the receptor containing the γ1 subunit was determined by competition for 3 H]RO7239181 (67.3 Ci / mmol; Roche) binding to membranes from HEK293F cells (ThermoFisher R79007) expressing human (transiently transfected) receptors of the composition α5β2γ1, α2β2γ1, α1β2γ1. For better protein expression of the receptor containing the α2 subunit, human GABA A The 28-amino acid long signal peptide (Met1 - Ala28) of the γ2 subunit was replaced with A the 31-amino acid long signal peptide (Met1 - Ser31) of the α5 subunit.

[0073] Various GABA APellets collected from HEK293F cells expressing the receptor subtype were resuspended in mannitol buffer pH 7.2 - 7.4 (mannitol 0.29 M, triethylamine 10 mM, acetic acid 10 mM, 1 mM EDTA + protease inhibitor (20 tablets Complete, Roche Diagnostics catalog No. 05 056 489 001 / liter)), washed twice, and then resuspended in the same buffer at a dilution of 1:10 - 1:15. Cell disruption was carried out by stirring the suspension at 435 psi for 15 minutes in a Parr vessel #4637, and then the suspension was centrifuged at 4°C, 1000 x g for 15 minutes (Beckman Avanti J-HC; rotor JS-4.2). The supernatant (S1) was transferred to a 2 l Schott flask, and the pellet (P1) was resuspended to 175 ml with mannitol buffer. The resuspended pellet was transferred to a 250 ml Corning centrifuge beaker and centrifuged at 4°C, 1500 x g for 10 minutes (Beckman Avanti J-HC; rotor JS-4.2). Then, the supernatant (S1) was transferred to a 2 l Schott flask and the pellet was discarded. The supernatant (S1) was centrifuged at 4°C, 15,000 x g for 30 minutes in a 500 ml Beckman polypropylene centrifuge beaker (Beckman Avanti J-20 XP; rotor JLA-10.500). The pellet (P2) was resuspended 1:1 with mannitol buffer and frozen at -80°C. The supernatant (S2) was centrifuged at 4°C, 48000 x g for 50 minutes in a 100 ml Beckman polypropylene centrifuge tube (Beckman Avanti J-20 XP; rotor JA-18). The supernatant (S3) was discarded and the pellet (P3) was resuspended 1:1 with mannitol buffer. The protein concentrations of P2 and P3 were measured using the BIORAD Standard assay method using bovine serum albumin as a standard and measured with a NANO-Drop 1000. A fixed amount of the membrane suspension was transferred to tubes (500 μl / tube) and stored at -80°C until needed.

[0074] Membrane homogenates were resuspended and polytronized (Polytron PT1200 E Kinematica AG) in 10 mM potassium phosphate, 100 mM KCl binding buffer, pH 7.4, to final assay concentrations determined in previous experiments.

[0075] Radioligand binding assays were performed using 100 μL of cell membranes and 1.5 nM (α5β2γ1) or 20–30 nM (α1β2γ1, α2β2γ1) concentrations of [ 3 H]RO7239181, and [0.3–10000] × 10 -9 The assay was performed in a volume of 200 μL (96-well plate) containing test compounds in the range of M. Non-specific binding was determined to be 10×10 -6 (α5β2γ1) and 30×10 -6 The binding was defined by MRO7235136 and typically represented less than 5% (α5β2γ1) and 20% (α1β2γ1,α2β2γ1) of the total binding. Assays were incubated at 4°C for 1 h to equilibrium, then membranes were filtered onto 96-well white microplates with GF / C filters preincubated for 20-50 min in Unifil (0.3% polyethyleneimine) using a Filtermate 196 harvester (Packard BioScience) and washed four times with cold potassium phosphate 10 mM pH 7.4, KCl 100 mM binding buffer. After drying, radioactivity retained on the filters was detected by liquid scintillation counting. K i Values ​​were calculated using Excel-Fit (Microsoft) and are the average of duplicate determinations.

[0076] The compounds of the accompanying examples were tested in the above assay and preferred compounds were A [From γ1 subunit-containing receptors (e.g., α5β2γ1, α2β2γ1, α1β2γ1) 3 K of 100 nM or less for displacement of H]RO7239181 iIt was found to have a value. Compounds with Ki (nM) < 50 are most preferred. Table 1 shows representative test results obtained by the above assay for measuring the binding affinity to HEK293 cells expressing the human (h) receptor.

[0077] [3 H Preparation of RO7239181, 6-chloro-5-(2,6-difluorophenyl)-7-methyl-1-(tritritiomethyl)-3H-1,4-benzodiazepin-2-one

Chemical formula

[0078] a) 6-chloro-5-(2,6-difluorophenyl)-7-methyl-1,3-dihydro-1,4-benzodiazepin-2-one Into a microwave tube, 7-bromo-6-chloro-5-(2,6-difluorophenyl)-1,3-dihydro-1,4-benzodiazepin-2-one (constituent unit A (see below), 450 mg, 1.17 mmol), trimethylboroxin (205 mg, 228 μL, 1.63 mmol), potassium carbonate (242 mg, 1.75 mmol) and tetrakis(triphenylphosphine)palladium(0) (67.4 mg, 58.4 μmol) were charged. Degassed 1,4-dioxane (8.1 mL) and H2O (2.7 ml) were added, and then the vial was capped. The suspension was reacted in a microwave at 130 °C for 30 minutes to be completely converted. The mixture was evaporated, treated with saturated NaHCO3 solution (20 ml), and extracted with EtOAc (2 × 20 mL). The organic layer was dried over Na2SO4, filtered, and the solvent was evaporated. The residue was purified by flash chromatography (silica gel, 40 g, eluting with CH2Cl2 / EtOAc 10% - 40% - 70% in heptane) to obtain the title compound (344 mg, 92%) as a pale yellow solid. MS(ESI): 321.1([M + H] + ).

[0079] b) 6-Chloro-5-(2,6-difluorophenyl)-7-methyl-1-(tritiomethyl)-3H-1,4-benzodiazepin-2-one In THF (200 μL) 3 To a solution of [H]methyl nosylate (1.85 GBq, 50 mCi, 0.61 μmol) in THF (200 μL) was added N-desmethyl precursor 6-chloro-5-(2,6-difluorophenyl)-7-methyl-1,3-dihydro-1,4-benzodiazepin-2-one (0.43 mg, 1.34 μmol) dissolved in THF (200 μL) and 10 equivalents of sodium tert-butyrate (0.5 M in THF, 13.4 μmol). After stirring at room temperature for 4 h, the reaction mixture was treated with H2O, evaporated, and the crude product was purified by HPLC (X-Terra Prep RP-18, 10 × 150 mm, MeCN / H2O (containing 5% MeCN) 40:60, 4 mL / min, 230 nm). The pure tritium-labeled compound was isolated by solid-phase extraction (Sep-Pak Plus C18), eluted from the cartridge as an ethanol solution, and a target compound with >99% radiochemical purity of 1.6 GBq (43.2 mCi) was obtained. The specific activity determined by mass spectrometry (MS) was 2.49 TBq / mmol (67.3 Ci / mmol). The identity of the labeled compound was confirmed by HPLC (by co-injecting an unlabeled reference standard) and MS.

[0080] MS: m / z = 335 [M(H)+H] + (16%), 337 [M( 3 H)+H] + (0%), 339 [M( 3 H2)+H] + (16%), 341 [M( 3 H3)+H] + (68%).

[0081] γ2-containing GABA A Membrane preparation and binding assay for the subtype GABA A The affinity of the compound for the γ2 subunit-containing receptor was determined by binding to HEK293F cells expressing the human (transiently transfected) receptor of composition α1β3γ23 Measured by competition for [³H]flumazenil (81.1 Ci / mmol; Roche) binding.

[0082] Various GABA A Pellets collected from HEK293F cells expressing the γ2 receptor subtype were resuspended in mannitol buffer pH 7.2 - 7.4, and GABA A Cells expressing the γ1 subunit-containing receptor were treated as described above.

[0083] The radioligand binding assay was performed in a volume of 200 μL (96-well plate) containing 100 μL of cell membranes, [³H]flumazenil at a concentration of 1 nM, and test compounds in the range of 0.1×10 3 to 30×10 -9 ~30×10 -6 M. Nonspecific binding was defined by 10 -5 M diazepam and was usually less than 5% of total binding. The assay was incubated for 1 hour at 4 °C for equilibrium, filtered using a Packard harvester, and harvested onto GF / C unifilters (Packard) by washing with ice-cold wash buffer (50 mM Tris, pH 7.5). After drying, radioactivity retained on the filter was detected by liquid scintillation counting. K i values were calculated using Excel-Fit (Microsoft) and are the average of two measurements.

[0084] The compounds of the appended examples were tested in the above assay, and preferred compounds had high K A values for displacement of [³H]flumazenil from the α1β3γ2 subtype of the human GABA 3 receptor. Compounds with K i α1β3γ2 (nM) > 300 were most preferred. In a preferred embodiment, the compounds of the present invention are binding selective for the γ2 subunit-containing GABA i receptor over the γ1 subunit-containing GABA A receptor. In particular, the compounds of the present invention have a "K A greater than 10-fold. iα1β3γ2 (nM) / K i The selection ratio of γ2 / γ1 defined as "α2β2γ1 (nM)", or "Log[K i α1β3γ2 (nM) / K i Representative test results obtained by the above assay for measuring the binding affinity to HEK293 cells expressing a human (h) receptor having LogSel defined as "α2β2γ1 (nM)]" are shown in Table 1 below.

[0085]

Table 1

[0086] GABA A Functional expression of the receptor: Preparation of Xenopus laevis oocytes Xenopus laevis oocytes at maturation stages V - VI were used for the expression of cloned mRNA encoding the GABA A receptor subunit. Oocytes ready for RNA microinjection were purchased from Ecocyte, Castrop - Rauxel, Germany, and stored at 20 °C in modified Barth medium (composition in mM: NaCl 88, KCl 1, NaHCO3 2.4, HEPES 10, MgSO4 0.82, CaNO3 0.33, CaCl2 0.33, pH = 7.5) until the experiment.

[0087] Xenopus laevis oocyte microinjection Oocytes were seeded into 96 - well plates for microinjection using a Roboinject automated device (MultiChannelSystems, Reutlingen, Germany). The desired GABA AApproximately 50 nL of an aqueous solution containing RNA transcripts of the subunits of the receptor subtype was injected into each oocyte. The RNA concentration ranged between 20 pg / μL / subunit and 200 pg / μL / subunit and was adjusted in pilot experiments to obtain an appropriate size and maximal effect of GABA A responses at the GABA receptor benzodiazepine (BZD) binding site with the reference benzodiazepine positive allosteric modulators (PAMs) flunitrazepam, triazolam, and midazolam. Oocytes were stored in modified Barth medium at 20 °C (composition in mM: NaCl 88, KCl 1, NaHCO3 4, HEPES 10, MgSO4 0.82, CaNO3 0.33, CaCl2 0.33, pH = 7.5) until the experiment.

[0088] Electrophysiology Electrophysiological experiments were performed on days 3 - 5 after microinjection of mRNA using a Roboocyte apparatus (MultiChannelSystems, Reutlingen, Germany). During the experiment, oocytes were constantly superfused with a solution containing (in mM) NaCl 90, KCl 1, HEPES 5, MgCl2 1, CaCl2 1 (pH 7.4). Oocytes were impaled with two glass microelectrodes (resistance: 0.5 - 0.8 MΩ) filled with a solution containing 1 M KCl + 1.5 M K-acetate and voltage clamped at -80 mV. Recordings were made at room temperature using a Robocyte two-electrode voltage clamp system (Multichannelsystem). After an initial equilibration period of 1.5 min with GABA, it was added for 1.5 min at a concentration that evoked approximately 20% of the maximal current response (EC 20 ). Another rest interval of 2.5 min with GABA was added again, after which a response of the same amplitude and shape was elicited. 0.5 min after the start of this second GABA application, while GABA was still present, a test compound at a concentration corresponding to approximately 30-fold that of K i α2β2γ1 was added. Current traces were recorded at a digitization rate of 10 Hz during and around GABA application.

[0089] Each compound and concentration was tested on at least 3 oocytes. Different oocytes were used for different compound concentrations. Reference PAM, flunitrazepam, triazolam and midazolam enhanced the GABA-induced current in the α2β2γ1 GABA A receptor subtype by approximately 60%.

[0090] Data analysis For analysis, the digitized current traces of the first and second GABA responses were overlaid and, if necessary, rescaled to equal maximum amplitude. The ratio between the two responses during the time interval of the test compound experiment was calculated point by point. The extreme values of the resulting "ratio trace" were taken as the efficacy ("fold increase") of the compound expressed as "% modulation of GABA EC 20 " (100*(fold increase - 1)).

[0091] The results are shown in Table 2.

Table 2

[0092] (Reference compound) The reference compounds (classical commercially available benzodiazepines) listed below and their structural analogs were also tested for affinity for the GABA A receptor subtypes α1β2γ1 and α2β2γ1 and for the GABA A receptor subtype α1β3γ2. The results are shown in Table 3.

Chemical formula

[0093]

Table 3

[0094] The compounds of formula (I) and their pharmaceutically acceptable salts can be used as pharmaceuticals (e.g., in the form of pharmaceutical preparations). The pharmaceutical preparations of the present invention can be administered orally (e.g., in the form of tablets, coated tablets, dragees, hard and soft gelatin capsules, solutions, emulsions or suspensions), nasally (e.g., in the form of nasal drops), rectally (e.g., in the form of suppositories), or topically to the eye (e.g., in the form of solutions, ointments, gels or water-soluble polymer inserts). However, administration can also be carried out parenterally, such as intramuscularly, intravenously, or intravitreally (e.g., in the form of a sterile injectable solution).

[0095] The compounds of formula (I) and their pharmaceutically acceptable salts can be processed with pharmaceutically inert, inorganic or organic adjuvants for the manufacture of tablets, coated tablets, dragees, hard gelatin capsules, injection solutions or topical agents. Lactose, corn starch or its derivatives, talc, stearic acid or its salts, etc. can be used, for example, as such adjuvants for tablets, dragees and hard gelatin capsules.

[0096] Examples of adjuvants suitable for soft gelatin capsules include vegetable oils, waxes, fats, semi-solids, liquid polyols, and the like.

[0097] Adjuvants suitable for the production of solutions and syrups are, for example, water, polyols, sucrose, invert sugar, glucose, and the like.

[0098] Adjuvants suitable for injection solutions are, for example, water, alcohol, polyols, glycerol, vegetable oils, and the like.

[0099] Adjuvants suitable for suppositories are, for example, natural or hardened oils, waxes, fats, semi-solids or liquid polyols, and the like.

[0100] Suitable adjuvants for ophthalmic topical forms are, for example, cyclodextrins, mannitol, or many other carriers and excipients known in the art.

[0101] Furthermore, the pharmaceutical preparation can contain preservatives, solubilizers, viscosity increasing substances, stabilizers, wetting agents, emulsifiers, sweeteners, coloring agents, flavoring agents, salts for changing osmotic pressure, buffering agents, masking agents, or antioxidants. It can also further contain other therapeutically useful substances.

[0102] The dosage can vary widely and of course should be adapted to the individual requirements in each particular case. Generally, for oral administration, about 0.1 mg to 20 mg per kg of body weight, preferably about 0.5 mg to 4 mg per kg of body weight (for example, about 300 mg per person) per day, is preferably administered individually in 1 to 3 divided doses, and if appropriate, can be constituted, for example, by the same amount. In the case of topical administration, the preparation can contain 0.001 wt% to 15 wt% of the medicine, and the required amount can be between 0.1 and 25 mg, and can be a single administration per day, a single administration per week, multiple administrations per day (2 to 4 times), or multiple administrations per week. However, it is obvious that there may be cases where the upper or lower limits described here are exceeded.

[0103] Preparation of a pharmaceutical composition comprising the compound of the present invention Tablets of the following composition are manufactured by a conventional method: [Table 4]

[0104] Manufacturing procedure 1. Mix components 1, 2, 3, and 4 and granulate with purified water. 2. Dry the granules at 50°C. 3. Pass the granules through an appropriate grinding device. 4. Add material 5, mix for 3 minutes, and compress with an appropriate press.

[0105] Manufacture capsules of the following composition: [Table 5]

[0106] Manufacturing procedure 1. Mix materials 1, 2, and 3 in a suitable mixer for 30 minutes. 2. Add materials 4 and 5 and mix for 3 minutes. 3. Fill into appropriate capsules.

[0107] The compound of formula I, lactose, and corn starch are first mixed in a mixer and then in a grinder. The mixture is returned to the mixer, and talc is added thereto and mixed uniformly. This mixture is filled into appropriate capsules, for example, hard gelatin capsules, by a machine.

[0108] Prepare an injection solution with the following composition. [Table 6]

[0109] Hereinafter, the present invention will be described by way of examples, but the present invention is not limited thereto.

[0110] When the fractionation example is obtained as a mixture of enantiomers, the pure enantiomers can be obtained by the methods described herein or methods known to those skilled in the art, for example, by chiral chromatography or crystallization.

[0111] Example Component A 7-Bromo-6-chloro-5-(2,6-difluorophenyl)-1,3-dihydro-1,4-benzodiazepin-2-one [Chemical formula]

[0112] a) 5-Chloro-2-methyl-3,1-benzoxazin-4-one A solution of 2-amino-6-chlorobenzoic acid (250 g, 1.46 mol) in acetic anhydride (1250 mL) was stirred at 140 °C for 2 h. The reaction mixture was concentrated under reduced pressure. The obtained crude residue was suspended in ethyl acetate (1000 mL), stirred for 30 min, filtered, and dried under reduced pressure to obtain the title compound (238 g, 84%) as a gray solid. 1 H NMR (DMSO-d6, 400 MHz): δ: 7.80 (app t, J = 8.0 Hz, 1H), 7.62 (d, J = 8.0 Hz, 1H), 7.49 (d, J = 7.6 Hz, 1H), 2.36 (s, 3H).

[0113] b) N-[3-chloro-2-(2,6-difluorobenzoyl)phenyl]acetamide To a solution of 5-chloro-2-methyl-3,1-benzoxazin-4-one (100 g, 511.2 mmol) and 2-bromo-1,3-difluorobenzene (118.4 g, 613.5 mmol) in tetrahydrofuran (1000 mL) was added dropwise i-PrMgCl·LiCl (1.3 m, 500 mL, 650 mmol) at -70 °C under nitrogen. The mixture was warmed to room temperature within 1 h, the reaction was quenched with saturated aqueous ammonium chloride solution (1500 mL), and extracted with ethyl acetate (2 × 1500 mL). The organic phase was washed with brine (2000 mL), dried over sodium sulfate, and concentrated under reduced pressure. The residue was suspended in ethyl acetate (150 mL). The obtained suspension was stirred at room temperature for 20 min, filtered, and dried under reduced pressure to obtain the title compound (113 g, 71%) as an off-white solid. 1 H NMR (DMSO-d6, 400 MHz): δ: 9.85 (s, 1H), 7.65 - 7.45 (m, 1H), 7.40 (t, J = 7.2 Hz, 1H), 7.38 - 7.34 (m, 2H), 7.16 (t, J = 8.8 Hz, 2H), 1.85 (s, 3H).

[0114] c) (2-Amino-6-chloro-phenyl)-(2,6-difluorophenyl)methanone To a solution of N-[3-chloro-2-(2,6-difluorobenzoyl)phenyl]acetamide (113 g, 364.9 mmol) in ethanol (250 mL) was added aqueous hydrochloric acid (12 m, 200 mL). The reaction mixture was stirred at 100 °C for 1 hour and then diluted with ethyl acetate (1100 mL). The organic phase was washed with water (1100 mL), saturated aqueous sodium bicarbonate (1100 mL) and brine (1100 mL), dried over sodium sulfate and concentrated under reduced pressure. Petroleum ether (120 mL) was added to the crude product and the suspension was stirred at room temperature for 20 minutes. The solid was filtered off and dried to give the title compound (88 g, 90%) as a yellow solid. 1 H NMR (DMSO-d6, 400 MHz): δ: 7.62 - 7.56 (m, 1H), 7.21 - 7.15 (m, 3H), 6.83 (d, J = 7.6 Hz, 1H), 6.74 (s, 2H), 6.58 (d, J = 7.6 Hz, 1H).

[0115] d) (6-Amino-3-bromo-2-chloro-phenyl)-(2,6-difluorophenyl)methanone (2-Amino-6-chloro-phenyl)-(2,6-difluorophenyl)methanone (88.0 g, 328.8 mmol) in dichloromethane (225 mL) and N,N-dimethylformamide (225 mL) was added 1-bromopyrrolidine-2,5-dione (64.4 g, 362 mmol) at 0 °C. The reaction mixture was stirred at 30 °C for 1 hour. The mixture was diluted with dichloromethane (600 mL), washed with water (500 mL) and brine (4 × 500 mL), dried over sodium sulfate and concentrated under reduced pressure. The residue was purified by chromatography (silica, petroleum ether / ethyl acetate, 1:0~2:1). The solid was suspended in petroleum ether (200 mL) and stirred at room temperature for 20 minutes. The suspension was filtered and the solid was dried under reduced pressure to give the title compound (96.0 g, 84%) as a yellow solid. MS: 345.9 ([{ 79 Br, 35 Cl}M+H] + ), 347.8 ([{ 81 Br, 35 Cl or 79 Br,37 Cl}M+H] + ), ESI pos.

[0116] e) 7-Bromo-6-chloro-5-(2,6-difluorophenyl)-1,3-dihydro-1,4-benzodiazepin-2-one (6-Amino-3-bromo-2-chloro-phenyl)-(2,6-difluorophenyl)methanone (25.0 g, 72.1 mmol) in pyridine (625 mL) was added ethyl 2-aminoacetate hydrochloride (70.5 g, 505 mmol). The reaction mixture was stirred at 135 °C for 36 h. The reaction mixture was concentrated under reduced pressure to remove pyridine. The residue was diluted with ethyl acetate (2000 mL) and washed with HCl (1.0 m, 3 × 1500 mL), water (2000 mL) and brine (2 × 1000 mL), dried over sodium sulfate, filtered and concentrated under reduced pressure. The crude product was purified by flash column chromatography (silica, petroleum ether / ethyl acetate 10:1 - 2:1) to give the title compound (10.1 g, 12%) as an off-white solid. MS: 385.0 ([{ 79 Br, 35 Cl}M+H] + ), ESI pos.

[0117] Component B 7-Bromo-6-chloro-5-(2-fluorophenyl)-1,3-dihydro-1,4-benzodiazepin-2-one

Chemical formula

[0118] a) N-[3-Chloro-2-(2-fluorobenzoyl)phenyl]acetamide A solution of 5-chloro-2-methyl-3,1-benzoxazin-4-one (20.0 g, 102.3 mmol) and 1-bromo-2-fluorobenzene (17.9 g, 102.3 mmol) in tetrahydrofuran (600 mL) was added dropwise with n-BuLi (2.5 M, 49 mL, 123 mmol) in tetrahydrofuran at -70 °C. The reaction mixture was stirred at -60 °C for 1 hour and then the reaction was quenched with aqueous ammonium chloride solution (200 mL). The aqueous layer was extracted with tetrahydrofuran (2 × 250 mL) and ethyl acetate (2 × 250 mL). The combined organic phases were washed with brine (200 mL), dried over sodium sulfate and concentrated under reduced pressure. Purification by flash column chromatography (silica, petroleum ether / ethyl acetate 20:1 - 3:1) gave the title compound (21 g, 70%) as a white solid. MS: 292.3 ([M+H] + ), ESI pos.

[0119] b) (2-Amino-6-chloro-phenyl)-(2-fluorophenyl)methanone In the same experiment as for component Ac, N-[3-chloro-2-(2-fluorobenzoyl)phenyl]acetamide was converted to the title compound (10 g, 58%) and obtained as a yellow solid. MS: 250.1 ([M+H] + ), ESI pos.

[0120] c) (6-Amino-3-bromo-2-chloro-phenyl)-(2-fluorophenyl)methanone In the same experiment as for component Ad, (2-amino-6-chloro-phenyl)-(2-fluorophenyl)methanone was converted to the title compound (32.4 g, 70%) and obtained as a yellow solid. MS: 327.9 ([{ 79 Br, 35 Cl}M+H] + ), 330.0 ([{ 81 Br, 35 Cl or 79 Br, 37 Cl}M+H] + ), ESI pos.

[0121] d) 7-Bromo-6-chloro-5-(2-fluorophenyl)-1,3-dihydro-1,4-benzodiazepin-2-one (6-Amino-3-bromo-2-chloro-phenyl)-(2-fluorophenyl)methanone (35.0 g, 98.3 mmol) in pyridine (210 mL) was added ethyl 2-aminoacetate hydrochloride (96.0 g, 688 mmol) at 90 °C. The reaction mixture was stirred at 110 °C for 16 h. The reaction mixture was cooled to room temperature and most of the pyridine was removed under reduced pressure. The residue was diluted with ethyl acetate (1250 mL). The organic phase was washed with aqueous HCl (1.0 m, 1250 mL), water (500 mL) and brine (1000 mL), dried over sodium sulfate, filtered and concentrated under reduced pressure. The crude product was purified by flash column chromatography (silica, petroleum ether / ethyl acetate, 1:0, 25:1, 1:1). The product was dissolved in ethyl acetate (15 mL). Petroleum ether (45 mL) was added dropwise to give a white slurry. The solid was collected by filtration and dried under reduced pressure to give the title compound (30.4 g, 39%) as an off-white solid. MS: 367.0 ([{ 79 Br, 35 Cl}M+H] + ), 368.9 ([{ 81 Br, 35 Cl or 79 Br, 37 Cl}M+H] + ), ESI pos.

[0122] Component G (rac)-7-Bromo-6-chloro-5-(2-fluorophenyl)-3-methyl-1,3-dihydro-1,4-benzodiazepin-2-one

Chemical formula

[0123] a) (rac)-tert-Butyl N-[2-[4-bromo-3-chloro-2-(2-fluorobenzoyl)anilino]-1-methyl-2-oxo-ethyl]carbamate (6-Amino-3-bromo-2-chloro-phenyl)-(2-fluorophenyl)methanone (2.0 g, 6.09 mmol) and 2-(tert-butoxycarbonylamino)propanoic acid (1.73 g, 9.13 mmol) in pyridine (20 mL) were slowly added with phosphoryl chloride (1.22 g, 7.98 mmol) at -5 °C. The reaction mixture was stirred at -5 °C for 1 hour, then slowly poured into water (200 mL) and extracted with ethyl acetate (2 × 100 mL). The combined organic phases were washed with brine (2 × 50 mL), dried over sodium sulfate and concentrated under reduced pressure. The residue was purified by flash column chromatography (petroleum ether / ethyl acetate 5:1) to give the title compound (2.95 g, 97%) as a yellow solid. MS: 399.1([{ 79 Br, 35 Cl}M-C4H8-CO2+H] + ), 401.0([{ 81 Br, 35 Cl or 79 Br, 37 Cl}M-C4H8-CO2+H] + ), ESI pos.

[0124] b) (rac)-2-Amino-N-[4-bromo-3-chloro-2-(2-fluorobenzoyl)phenyl]propanamide To a solution of (rac)-tert-butyl N-[2-[4-bromo-3-chloro-2-(2-fluorobenzoyl)anilino]-1-methyl-2-oxo-ethyl]carbamate (2.9 g, 5.8 mmol) in dichloromethane (14.5 mL) was slowly added hydrochloric acid (4.0 M in dioxane, 14.5 mL, 58.0 mmol). The reaction mixture was stirred at 25 °C for 2 hours. Saturated aqueous sodium hydrogen carbonate was slowly added until pH > 8, then the mixture was extracted with dichloromethane (3 × 100 mL). The combined organic phases were washed with brine (100 mL), dried over sodium sulfate and concentrated under reduced pressure to give the title compound (2.2 g, 92%) as a yellow oil. MS: 399.0([{ 79 Br, 35 Cl}M+H] + ), 401.0([{ 81Br, 35 Cl or 79 Br, 37 Cl}M+H] + ), ESI pos.

[0125] c)(rac)-7-bromo-6-chloro-5-(2-fluorophenyl)-3-methyl-1,3-dihydro-1,4-benzodiazepin-2-one (rac)-2-Amino-N-[4-bromo-3-chloro-2-(2-fluorobenzoyl)phenyl]propanamide (2.2 g, 5.5 mmol) in ethanol (20 mL) was added acetic acid (4 mL). The reaction mixture was stirred at 80 °C for 16 h and then concentrated under reduced pressure. The formed crystals were filtered, triturated with ethyl acetate (15 mL), then recovered by filtration, dried under reduced pressure to give the title compound (1.6 g, 76%) as a yellow solid. MS: 381.0 ([{ 79 Br, 35 Cl}M+H] + ), 383.0 ([{ 81 Br, 35 Cl or 79 Br, 37 Cl}M+H] + ), ESI pos.

[0126] Component L (3S)-7-bromo-6-chloro-5-(2,6-difluorophenyl)-3-methyl-1,3-dihydro-1,4-benzodiazepin-2-one

Chemical formula

[0127] a) tert-Butyl N-[(1S)-2-[4-bromo-3-chloro-2-(2,6-difluorobenzoyl)anilino]-1-methyl-2-oxoethyl]carbamate Similar to the experiment of component Ga, using (2S)-2-(tert-butoxycarbonylamino)propanoic acid, (6-amino-3-bromo-2-chloro-phenyl)-(2,6-difluorophenyl)methanone was converted into the labeled compound (1.50 g, 98%), which was obtained as a yellow solid. MS: 418.7([{ 81 Br, 35 Cl or 79 Br, 37 Cl}M-C4H8-CO2+H] + ), 540.7([{ 81 Br, 35 Cl or 79 Br, 37 Cl}M+Na] + ), ESI pos.

[0128] b) (2S)-2-Amino-N-[4-bromo-3-chloro-2-(2,6-difluorobenzoyl)phenyl]propanamide Similar to the experiment of component Gb, tert-butyl N-[(1S)-2-[4-bromo-3-chloro-2-(2,6-difluorobenzoyl)anilino]-1-methyl-2-oxoethyl]carbamate was converted into the labeled compound (1.1 g, 94%), obtained as a yellow oil, which was used directly in the next step without further characterization.

[0129] c) (3S)-7-Bromo-6-chloro-5-(2,6-difluorophenyl)-3-methyl-1,3-dihydro-1,4-benzodiazepin-2-one To a solution of (2S)-2-amino-N-[4-bromo-3-chloro-2-(2,6-difluorobenzoyl)phenyl]propanamide (960 mg, 2.30 mmol) in toluene (9.19 mL) was added silica (138 mg, 2.30 mmol). The reaction mixture was stirred at 90 °C for 15 h and then concentrated under reduced pressure. The residue was purified by flash column chromatography (petroleum ether / ethyl acetate 3:1) to give the labeled compound (920 mg, 95%) as a yellow solid. MS: 399.1([{ 79 Br, 35 Cl}M+H] +), 401.1 ([{ 81 Br, 35 Cl or 79 Br, 37 Cl}M+H] + ), ESI pos.

[0130] Component M (3S)-6,7-dichloro-5-(2,6-difluorophenyl)-3-methyl-1,3-dihydro-1,4-benzodiazepin-2-one

Chem.

[0131] a) tert-butyl N-[(1S)-2-[3,4-dichloro-2-(2,6-difluorobenzoyl)anilino]-1-methyl-2-oxoethyl]carbamate Similar to the experiment of Component Ga, (2S)-2-(tert-butoxycarbonylamino)propanoic acid was used to convert (6-amino-2,3-dichloro-phenyl)-(2,6-difluorophenyl)methanone into the title compound (5.0 g, 64%), which was obtained as a yellow foam. The crude product was used directly in the next step without further characterization.

[0132] b) (2S)-2-amino-N-[3,4-dichloro-2-(2,6-difluorobenzoyl)phenyl]propanamide Similar to the experiment of Component Gb, tert-butyl N-[(1S)-2-[3,4-dichloro-2-(2,6-difluorobenzoyl)anilino]-1-methyl-2-oxoethyl]carbamate was converted into the title compound (3.6 g, 91%), which was obtained as a yellow oil. MS: 373.0 ([{ 35 Cl, 35 Cl}M+H] + ), ESI pos.

[0133] c) (3S)-6,7-dichloro-5-(2,6-difluorophenyl)-3-methyl-1,3-dihydro-1,4-benzodiazepin-2-one Similar to the experiment of component Lc, (2S)-2-amino-N-[3,4-dichloro-2-(2,6-difluorobenzoyl)phenyl]propanamide was converted to the title compound (3.20 g, 93%), which was obtained as a yellow foam. MS: 355.0 ([{ 35 Cl, 35 Cl}M+H] + ), ESI pos.

[0134] Component O (3S)-6,7-Dichloro-5-(3-fluoro-2-pyridyl)-3-methyl-1,3-dihydro-1,4-benzodiazepin-2-one

Chemical Structure

[0135] a) tert-Butyl N-[(1S)-2-[3,4-dichloro-2-(3-fluoropyridine-2-carbonyl)anilino]-1-methyl-2-oxoethyl]carbamate Similar to the experiment of component Ga, (2S)-2-(tert-butoxycarbonylamino)propanoic acid was used to convert N-[3,4-dichloro-2-(3-fluoropyridine-2-carbonyl)phenyl]acetamide to the title compound (8.6 g, 67%), which was obtained as a white solid. MS: 456.0 ([{ 35 Cl, 35 Cl}M+H] + ), ESI pos.

[0136] b) (2S)-2-Amino-N-[3,4-dichloro-2-(3-fluoropyridine-2-carbonyl)phenyl]propanamide Similar to the experiment of component Gb, tert-Butyl N-[(1S)-2-[3,4-dichloro-2-(3-fluoropyridine-2-carbonyl)anilino]-1-methyl-2-oxoethyl]carbamate was converted to the title compound (6.6 g, 100%), which was obtained as a yellow solid. MS: 356.0 ([{ 35 Cl, 35 Cl}M+H] + ), ESI pos.

[0137] c) (3S)-6,7-dichloro-5-(3-fluoro-2-pyridyl)-3-methyl-1,3-dihydro-1,4-benzodiazepin-2-one Similar to the experiment of component Lc, (2S)-2-amino-N-[3,4-dichloro-2-(3-fluoropyridine-2-carbonyl)phenyl]propanamide was converted to the labeled compound (5.5 g, 88%) and obtained as a yellow solid. MS: 338.0 ([{ 35 Cl, 35 Cl}M+H] + ), ESI pos.

[0138] Component Q (3S)-6-bromo-7-chloro-5-(2,6-difluorophenyl)-3-methyl-1,3-dihydro-1,4-benzodiazepin-2-one

Chemical formula

[0139] a) N-[3-bromo-4-chloro-2-(2,6-difluorobenzoyl)phenyl]acetamide Similar to the experiment of component Ad, 1-chloropyrrolidine-2,5-dione was used and N-(3-bromo-2-(2,6-difluorobenzoyl)phenyl)acetamide was converted to the labeled compound (10.1 g, 70%) and obtained as a pale yellow solid. MS: 388.0 ([{ 79 Br, 35 Cl}M+H] + ), 390.1 ([{ 81 Br, 35 Cl or 79 Br, 37 Cl}M+H] + ), ESI pos.

[0140] b) (6-amino-2-bromo-3-chloro-phenyl)-(2,6-difluorophenyl)methanone Similar to the experiment of Component Ac, N-[3-bromo-4-chloro-2-(2,6-difluorobenzoyl)phenyl]acetamide was converted to the title compound (8.2 g, 92%) and obtained as a yellow solid. MS: 346.0 ([{ 79 Br, 35 Cl}M+H] + ), 348.0 ([{ 81 Br, 35 Cl or 79 Br, 37 Cl}M+H] + ), ESI pos.

[0141] c) tert-Butyl N-[(1S)-2-[3-bromo-4-chloro-2-(2,6-difluorobenzoyl)anilino]-1-methyl-2-oxoethyl]carbamate Similar to the experiment of Component Ga, (2S)-2-(tert-butoxycarbonylamino)propanoic acid was used and (6-amino-2-bromo-3-chloro-phenyl)-(2,6-difluorophenyl)methanone was converted to the title compound (8.64 g, 69%) and obtained as a yellow solid. MS: 515.2 ([{ 79 Br, 35 Cl}M-H] - ), 517.1 ([{ 81 Br, 35 Cl or 79 Br, 37 Cl}M-H] - ), ESI neg.

[0142] d) (2S)-2-Amino-N-[3-bromo-4-chloro-2-(2,6-difluorobenzoyl)phenyl]propanamide Similar to the experiment of Component Gb, tert-Butyl N-[(1S)-2-[3-bromo-4-chloro-2-(2,6-difluorobenzoyl)anilino]-1-methyl-2-oxoethyl]carbamate was converted to the title compound (6.27 g, 90%) and obtained as a light brown oil. MS: 417.1 ([{ 79 Br, 35 Cl}M+H] + ), 419.0 ([{ 81 Br,35 Cl or 79 Br, 37 Cl}M+H] + ), ESI pos.

[0143] e) (3S)-6-bromo-7-chloro-5-(2,6-difluorophenyl)-3-methyl-1,3-dihydro-1,4-benzodiazepin-2-one Similar to the experiment of component Lc, (2S)-2-amino-N-[3-bromo-4-chloro-2-(2,6-difluorobenzoyl)phenyl]propanamide was converted to the labeled compound (3.98 g, 68%) and obtained as a yellow solid. MS: 399.1 ([{ 79 Br, 35 Cl}M+H] + ), 401.0 ([{ 81 Br, 35 Cl or 79 Br, 37 Cl}M+H] + ), ESI pos.

[0144] Component R (3S)-7-bromo-6-chloro-5-(3-fluoro-2-pyridyl)-3-methyl-1,3-dihydro-1,4-benzodiazepin-2-one

Chemical Structure

[0145] a) tert-butyl N-[(1S)-2-[4-bromo-3-chloro-2-(3-fluoropyridine-2-carbonyl)anilino]-1-methyl-2-oxo-ethyl]carbamate Similar to the experiment of component Ga, (2S)-2-(tert-butoxycarbonylamino)propanoic acid was used to convert (6-amino-3-bromo-2-chloro-phenyl)-(3-fluoro-2-pyridyl)methanone to the labeled compound (1.4 g, 97%) and obtained as a yellow foam. The unpurified product was used directly in the next step without further characterization.

[0146] b) (2S)-2-Amino-N-[4-bromo-3-chloro-2-(3-fluoropyridine-2-carbonyl)phenyl]propanamide Similar to the experiment of Component Gb, tert-butyl N-[(1S)-2-[4-bromo-3-chloro-2-(3-fluoropyridine-2-carbonyl)anilino]-1-methyl-2-oxo-ethyl]carbamate was converted to the title compound (1.1 g, 98%) and obtained as a yellow oil. The crude product was used directly in the next step without further characterization.

[0147] c) (3S)-7-Bromo-6-chloro-5-(3-fluoro-2-pyridyl)-3-methyl-1,3-dihydro-1,4-benzodiazepin-2-one Similar to the experiment of Component Lc, (2S)-2-Amino-N-[4-bromo-3-chloro-2-(3-fluoropyridine-2-carbonyl)phenyl]propanamide was converted to the title compound (430 mg, 40%) and obtained as a yellow solid. MS: 381.9 ([{ 79 Br, 35 Cl}M+H] + ), 383.9 ([{ 81 Br, 35 Cl or 79 Br, 37 Cl}M+H] + ), ESI pos.

[0148] Component S (3S)-6-Chloro-5-(2,6-difluorophenyl)-7-iodo-3-methyl-1,3-dihydro-1,4-benzodiazepin-2-one

Chemical Structure

[0149] a) tert-Butyl N-[(1S)-2-[3-chloro-2-(2,6-difluorobenzoyl)-4-iodo-anilino]-1-methyl-2-oxoethyl]carbamate Similar to the experiment of component Ga, using (2S)-2-(tert-butoxycarbonylamino)propanoic acid, (6-amino-2-chloro-3-iodo-phenyl)-(2,6-difluorophenyl)methanone was converted to the labeled compound (5.8 g, 81%) and obtained as a yellow solid. MS: 465.0([M-C4H8-CO2+H] + ), 509.0([M-C4H8+H] + ), ESI pos.

[0150] b) (2S)-2-Amino-N-[3-chloro-2-(2,6-difluorobenzoyl)-4-iodo-phenyl]propanamide Similar to the experiment of component Gb, tert-butyl N-[(1S)-2-[3-chloro-2-(2,6-difluorobenzoyl)-4-iodo-anilino]-1-methyl-2-oxoethyl]carbamate was converted to the labeled compound (4.7 g, 99%) and obtained as a yellow solid. The unpurified product was used directly in the next step without further characterization.

[0151] c) (3S)-6-Chloro-5-(2,6-difluorophenyl)-7-iodo-3-methyl-1,3-dihydro-1,4-benzodiazepin-2-one Similar to the experiment of component Lc, (2S)-2-amino-N-[3-chloro-2-(2,6-difluorobenzoyl)-4-iodo-phenyl]propanamide was converted to the labeled compound (3.8 g, 94%) and obtained as a yellow solid. MS: 446.8([M+H] + ), ESI pos.

[0152] Component T (3S)-7-Bromo-6-chloro-5-(2,6-difluorophenyl)-3-ethyl-1,3-dihydro-1,4-benzodiazepin-2-one

Chemical Structure

[0153] a) tert-Butyl N-[(1S)-1-[[4-bromo-3-chloro-2-(2,6-difluorobenzoyl)phenyl]carbamoyl]propyl]carbamate

[0154] Similar to the experiment of Component Ga, (2S)-2-(tert-butoxycarbonylamino)butanoic acid was used to convert (6-amino-3-bromo-2-chloro-phenyl)-(2,6-difluorophenyl)methanone into the labeled compound (1.08 g, 68%), which was obtained as a yellow solid. MS: 433.1 ([{ 81 Br, 35 Cl or 79 Br, 37 Cl}M-C4H8-CO2+H] + ), 477.1 ([{ 81 Br, 35 Cl or 79 Br, 37 Cl}M-C4H8+H] + ), ESI pos.

[0155] b) (2S)-2-Amino-N-[4-bromo-3-chloro-2-(2,6-difluorobenzoyl)phenyl]butanamide Similar to the experiment of Component Gb, tert-butyl N-[(1S)-1-[[4-bromo-3-chloro-2-(2,6-difluorobenzoyl)phenyl]carbamoyl]propyl]carbamate was converted into the labeled compound (730 mg, 90%), which was obtained as a yellow oil. MS: 433.0 ([{ 81 Br, 35 Cl or 79 Br, 37 Cl}M+H] + ), ESI pos.

[0156] c) (3S)-7-Bromo-6-chloro-5-(2,6-difluorophenyl)-3-ethyl-1,3-dihydro-1,4-benzodiazepin-2-one Similar to the experiment of Component Lc, (2S)-2-amino-N-[4-bromo-3-chloro-2-(2,6-difluorobenzoyl)phenyl]butanamide was converted to the title compound (650 mg, 97%) and obtained as a light brown foam. MS: 414.9([{ 81 Br, 35 Cl or 79 Br, 37 Cl}M+H] + ), ESI pos.

[0157] Component U (3S)-7-bromo-6-chloro-5-(2,6-difluorophenyl)-3-(methoxymethyl)-1,3-dihydro-1,4-benzodiazepin-2-one

Chemical Structure

[0158] a) tert-Butyl N-[(1S)-2-[4-bromo-3-chloro-2-(2,6-difluorobenzoyl)anilino]-1-(methoxymethyl)-2-oxoethyl]carbamate Similar to the experiment of Component Ga, (2S)-2-(tert-butoxycarbonylamino)-3-methoxy-propanoic acid was used to convert (6-amino-3-bromo-2-chloro-phenyl)-(2,6-difluorophenyl)methanone to the title compound (4.3 g, 85%) and obtained as a yellow solid. The crude product was used directly in the next step without further characterization.

[0159] b) (2S)-2-Amino-N-[4-bromo-3-chloro-2-(2,6-difluorobenzoyl)phenyl]-3-methoxy-propaneamide Similar to the experiment of Component Gb, tert-Butyl N-[(1S)-2-[4-bromo-3-chloro-2-(2,6-difluorobenzoyl)anilino]-1-(methoxymethyl)-2-oxoethyl]carbamate was converted to the title compound (3.0 g, 96%) and obtained as a yellow oil. MS: 447.0([{ 79 Br, 35Cl}M+H] + ),449.1([{ 81 Br, 35 Cl or 79 Br, 37 Cl}M+H] + ), ESI pos.

[0160] c) (3S)-7-Bromo-6-chloro-5-(2,6-difluorophenyl)-3-(methoxymethyl)-1,3-dihydro-1,4-benzodiazepin-2-one Similar to the experiment of Component Lc, (2S)-2-Amino-N-[4-bromo-3-chloro-2-(2,6-difluorobenzoyl)phenyl]-3-methoxy-propanamide was converted to the title compound (1.9 g, 78%) and obtained as a white solid. The crude product was used directly in the next step without further characterization. MS: 429.1([{ 79 Br, 35 Cl}M+H] + ), 431.1([{ 81 Br, 35 Cl or 79 Br, 37 Cl}M+H] + ), ESI pos.

[0161] Example 30 (4S)-8-Bromo-7-chloro-6-(2-fluorophenyl)-4-methyl-4H-[1,2,4]triazolo[4,3-a][1,4]benzodiazepine

Chemical Structure

[0162] a) (rac)-7-Bromo-6-chloro-5-(2-fluorophenyl)-3-methyl-1,3-dihydro-1,4-benzodiazepine-2-thione A suspension of (rac)-7-bromo-6-chloro-5-(2-fluorophenyl)-3-methyl-1,3-dihydro-1,4-benzodiazepin-2-one (Constituent G, 500 mg, 1.31 mmol) in toluene (8.33 mL) was added with Lawesson's reagent (635 mg, 1.57 mmol) at room temperature. The reaction mixture was stirred at 110 °C for 1 hour and then concentrated under reduced pressure. The crude purified substance was purified by flash column chromatography (silica, 15 - 25% ethyl acetate in heptane) to obtain the title compound (820 mg, 91%) as a yellow solid. The crude product was used directly in the next step without further characterization.

[0163] b) (rac)-8-Bromo-7-chloro-6-(2-fluorophenyl)-4-methyl-4H-[1,2,4]triazolo[4,3-a][1,4]benzodiazepine To a solution of (rac)-7-bromo-6-chloro-5-(2-fluorophenyl)-3-methyl-1,3-dihydro-1,4-benzodiazepine-2-thione (240 mg, 0.600 mmol) in butan-1-ol (3.2 mL) was added formohydroxamic acid (108 mg, 1.81 mmol). The reaction mixture was stirred at 120 °C for 16 hours. The mixture was diluted with dichloromethane (20 mL), washed with water (20 mL), brine (20 mL), dried over sodium sulfate, and concentrated under reduced pressure. The residue was purified by preparative HPLC (TFA) to obtain the title compound (250 mg, 100%) as a white solid. MS: 404.8 ([{ 79 Br, 35 Cl}M+H] + ), 406.7 ([{ 81 Br, 35 Cl or 79 Br, 37 Cl}M+H] + ), ESI pos.

[0164] c) (4S)-8-Bromo-7-chloro-6-(2-fluorophenyl)-4-methyl-4H-[1,2,4]triazolo[4,3-a][1,4]benzodiazepine (rac)-8-Bromo-7-chloro-6-(2-fluorophenyl)-4-methyl-4H-[1,2,4]triazolo[4,3-a][1,4]benzodiazepine (254 mg, 0.630 mmol) was purified by SFC (Chiralcel OJ-3, 0.05% diethylamine in methanol, 5 - 40%), The enantiopure (-)-(S)-labeled compound (88.1 mg) was obtained as a white solid. MS: 404.8 ([{ 79 Br, 35 Cl}M+H] + ), 406.8 ([{ 81 Br, 35 Cl or 79 Br, 37 Cl}M+H] + ), ESI pos. The enantiopure (+)-(R)-labeled compound (80.4 mg) was obtained as a white solid. MS: 404.9 ([{ 79 Br, 35 Cl}M+H] + ), 406.8 ([{ 81 Br, 35 Cl or 79 Br, 37 Cl}M+H] + ), ESI pos.

[0165] Example 31 8-Bromo-7-chloro-6-(2,6-difluorophenyl)-4H-[1,2,4]triazolo[4,3-a][1,4]benzodiazepin-4-ol

Chemical formula

[0166] a) 7-Bromo-6-chloro-5-(2,6-difluorophenyl)-1,3-dihydro-1,4-benzodiazepine-2-thione A suspension of 7-bromo-6-chloro-5-(2,6-difluorophenyl)-1,3-dihydro-1,4-benzodiazepin-2-one (Constituent A, 6.7 g, 17.4 mmol) in toluene (167 mL) was added with Lawesson's reagent (8.43 g, 20.9 mmol) at room temperature. After stirring the reaction mixture at 120 °C for 1.5 hours, it was diluted with ethyl acetate (400 mL). The organic layer was washed with water (300 mL) and brine (300 mL), dried over sodium sulfate, filtered, and concentrated under reduced pressure. The crude purified substance was purified by flash column chromatography (silica, 15 - 30% ethyl acetate in heptane) to obtain the title compound (6.98 g, 100%) as a yellow solid. MS: 400.8 ([{ 79 Br, 35 Cl}M+H] + ), 402.9 ([{ 81 Br, 35 Cl or 79 Br, 37 Cl}M+H] + ), ESI pos.

[0167] b) 8-Bromo-7-chloro-6-(2,6-difluorophenyl)-4H-[1,2,4]triazolo[4,3-a][1,4]benzodiazepine In the same experiment as in Example 30b, using formohydrazide, 7-bromo-6-chloro-5-(2,6-difluorophenyl)-1,3-dihydro-1,4-benzodiazepine-2-thione was converted to the title compound (2.4 g, 32%) and obtained as a white solid. MS: 409.1 ([{ 79 Br, 35 Cl}M+H] + ), 411.1 ([{ 81 Br, 35 Cl or 79 Br, 37 Cl}M+H] + ), ESI pos.

[0168] c) 8-Bromo-7-chloro-6-(2,6-difluorophenyl)-4H-[1,2,4]triazolo[4,3-a][1,4]benzodiazepin-4-ol

[0169] A solution of sodium bis(trimethylsilyl)amide (1.0 M in tetrahydrofuran, 0.37 mL, 0.370 mmol) in tetrahydrofuran (5 mL) was added with 8-bromo-7-chloro-6-(2,6-difluorophenyl)-4H-[1,2,4]triazolo[4,3-a][1,4]benzodiazepine (100.0 mg, 0.240 mmol) at -65 °C. The mixture was stirred at -65 °C for 30 minutes, and then 2-(benzenesulfonyl)-3-phenyl-oxaziridine (97 mg, 0.370 mmol) was added. After the reaction mixture was stirred at -65 °C for 2 hours, the reaction was quenched by the addition of saturated aqueous ammonium chloride. The aqueous phase was extracted with ethyl acetate (2 × 10 mL). The combined organic layers were washed with brine (2 × 30 mL), filtered, and concentrated under reduced pressure. The residue was purified by preparative HPLC (BostonPrime C18, 0.1% trifluoroacetic acid / water / acetonitrile), followed by chiral HPLC (Daicel Chiralcel OJ-H, methanol) to give the title compound (30 mg, 29%) as a white solid. MS: 424.9 ([{ 79 Br, 35 Cl}M+H] + ), 426.8 ([{ 81 Br, 35 Cl or 79 Br, 37 Cl}M+H] + ), ESI pos.

[0170] Example 32 (4S)-8-Bromo-7-chloro-6-(2-fluorophenyl)-4-methyl-1-pyridazin-3-yl-4H-[1,2,4]triazolo[4,3-a][1,4]benzodiazepine

Chemical Structure

[0171] a) (rac)-8-Bromo-7-chloro-6-(2-fluorophenyl)-4-methyl-1-pyridazin-3-yl-4H-[1,2,4]triazolo[4,3-a][1,4]benzodiazepine Similar to the experiment of Example 30b, using pyridazine-3-carbohydrazide, (rac)-7-bromo-6-chloro-5-(2-fluorophenyl)-3-methyl-1,3-dihydro-1,4-benzodiazepine-2-thione was converted to the labeled compound (220 mg, 75%) and obtained as a white solid. MS: 482.9 ([{ 79 Br, 35 Cl}M+H] + ), 484.8 ([{ 81 Br, 35 Cl or 79 Br, 37 Cl}M+H] + ), ESI pos.

[0172] b) (4S)-8-Bromo-7-chloro-6-(2-fluorophenyl)-4-methyl-1-pyridazin-3-yl-4H-[1,2,4]triazolo[4,3-a][1,4]benzodiazepine (rac)-8-Bromo-7-chloro-6-(2-fluorophenyl)-4-methyl-1-pyridazin-3-yl-4H-[1,2,4]triazolo[4,3-a][1,4]benzodiazepine (220 mg, 0.450 mmol) was purified by SFC (Chiralcel OJ-3, 0.05% diethylamine in methanol, 5 - 40%) and then lyophilized to obtain the following: Enantiopure (-)-(S)-labeled compound (64.3 mg) was obtained as a white solid. MS: 483.1 ([{ 79 Br, 35 Cl}M+H] + ), 485.1 ([{ 81 Br, 35 Cl or 79 Br, 37 Cl}M+H] + ), ESI pos. Enantiopure (+)-(R)-labeled compound (61.4 mg) was obtained as a white solid. MS: 483.1 ([{79 Br, 35 Cl}M+H] + ),485.1([{ 81 Br, 35 Cl or 79 Br, 37 Cl}M+H] + ), ESI pos.

[0173] Example 35 8-Bromo-7-chloro-6-(2-fluorophenyl)-1-pyridazin-3-yl-4H-[1,2,4]triazolo[4,3-a][1,4]benzodiazepin-4-ol

Chemical Structure

[0174] a) 7-Bromo-6-chloro-5-(2-fluorophenyl)-1,3-dihydro-1,4-benzodiazepine-2-thione

[0175] Similar to the experiment of Example 30a, 7-bromo-6-chloro-5-(2-fluorophenyl)-1,3-dihydro-1,4-benzodiazepine-2-one was converted to the title compound (390 mg, 54%) and obtained as a yellow solid. The crude product was used directly in the next step without further characterization.

[0176] b) 8-Bromo-7-chloro-6-(2-fluorophenyl)-1-pyridazin-3-yl-4H-[1,2,4]triazolo[4,3-a][1,4]benzodiazepine Similar to the experiment of Example 30b, pyridazine-3-carbohydrazide was used to convert 7-bromo-6-chloro-5-(2-fluorophenyl)-1,3-dihydro-1,4-benzodiazepine-2-thione to the title compound (294 mg, 73%) and obtained as a white solid. MS: 469.1([{ 79 Br, 35 Cl}M+H] + ), 471.1([{ 81 Br, 35 Cl or 79 Br,37 Cl}M+H] + ), ESI pos.

[0177] c)[8-Bromo-7-chloro-6-(2-fluorophenyl)-1-pyridazin-3-yl-4H-[1,2,4]triazolo[4,3-a][1,4]benzodiazepin-4-yl]acetate

[0178] To a solution of 8-bromo-7-chloro-6-(2-fluorophenyl)-1-pyridazin-3-yl-4H-[1,2,4]triazolo[4,3-a][1,4]benzodiazepine (100 mg, 0.21 mmol) in acetic acid (2 mL) were added iodine (27 mg, 0.11 mmol), potassium acetate (42 mg, 0.43 mmol) and potassium persulfate (58 mg, 0.21 mmol). The reaction mixture was heated at 90 °C for 12 h, and then the reaction was stopped by the addition of saturated aqueous Na2SO3 solution (10 mL). The mixture was extracted with ethyl acetate (30 mL). The organic layer was washed with water (10 mL), dried over sodium sulfate and concentrated under reduced pressure. The residue was purified by flash column chromatography (silica, dichloromethane / methanol 100:1 - 80:1) to give the title compound (55 mg, 49%) as a white solid. MS: 526.9([{ 79 Br, 35 Cl}M+H] + ), 528.9([{ 81 Br, 35 Cl or 79 Br, 37 Cl}M+H] + ), ESI pos.

[0179] d) 8-Bromo-7-chloro-6-(2-fluorophenyl)-1-pyridazin-3-yl-4H-[1,2,4]triazolo[4,3-a][1,4]benzodiazepin-4-ol

[0180] To a solution of [[8-bromo-7-chloro-6-(2-fluorophenyl)-1-pyridazin-3-yl-4H-[1,2,4]triazolo[4,3-a][1,4]benzodiazepin-4-yl]acetate (55 mg, 0.10 mmol) in ethanol (2.75 mL) was added sodium carbonate (22 mg, 0.21 mmol). The reaction mixture was stirred at room temperature for 4 h, then filtered and concentrated under reduced pressure. The residue was purified by preparative TLC (dichloromethane / methanol), then preparative HPLC (Shim-pack C18 in water / acetonitrile, 0.225% trifluoroacetic acid), and lyophilized to give the title compound (13.1 mg, 26%) as a white solid. MS: 485.0 ([{ 79 Br, 35 Cl}M+H] + ), 487.0 ([{ 81 Br, 35 Cl or 79 Br, 37 Cl}M+H] + ), ESI pos.

[0181] Example 51 (4S)-8-Bromo-7-chloro-6-(2,6-difluorophenyl)-4-methyl-4H-[1,2,4]triazolo[4,3-a][1,4]benzodiazepine

Chemical formula

[0182] a) (3S)-7-Bromo-6-chloro-5-(2,6-difluorophenyl)-3-methyl-1,3-dihydro-1,4-benzodiazepine-2-thione In the same experiment as in Example 30a, (3S)-7-bromo-6-chloro-5-(2,6-difluorophenyl)-3-methyl-1,3-dihydro-1,4-benzodiazepine-2-one (Component L) was converted to the title compound (410 mg, 96%) and obtained as a yellow solid. MS: 415.1 ([{ 79 Br, 35 Cl}M+H] + ), 417.0 ([{ 81 Br,35 Cl or 79 Br, 37 Cl}M+H] + ), ESI pos.

[0183] b) (4S)-8-bromo-7-chloro-6-(2,6-difluorophenyl)-4-methyl-4H-[1,2,4]triazolo[4,3-a][1,4]benzodiazepine

[0184] Similar to the experiment of Example 30b, using formohydrazide, (3S)-7-bromo-6-chloro-5-(2,6-difluorophenyl)-3-methyl-1,3-dihydro-1,4-benzodiazepine-2-thione was converted to the (-)-enantiopure (S)-labeled compound (66 mg, 43%) after chiral purification, and this was obtained as a white solid. MS: 423.0 ([{ 79 Br, 35 Cl}M+H] + ), 425.1 ([{ 81 Br, 35 Cl or 79 Br, 37 Cl}M+H] + ), ESI pos.

[0185] Example 52 (4S)-7,8-dichloro-6-(2,6-difluorophenyl)-4-methyl-1-pyrimidin-4-yl-4H-[1,2,4]triazolo[4,3-a][1,4]benzodiazepine

Chemical Structure

[0186] a) (3S)-6,7-dichloro-5-(2,6-difluorophenyl)-3-methyl-1,3-dihydro-1,4-benzodiazepine-2-thione

[0187] Similar to the experiment of Example 30a, (3S)-6,7-dichloro-5-(2,6-difluorophenyl)-3-methyl-1,3-dihydro-1,4-benzodiazepin-2-one (Component M) was converted to the labeled compound (4.2 g, 90%) and obtained as a yellow solid. The unpurified product was used as it was in the next step without further characterization.

[0188] b) (4S)-7,8-dichloro-6-(2,6-difluorophenyl)-4-methyl-1-pyrimidin-4-yl-4H-[1,2,4]triazolo[4,3-a][1,4]benzodiazepine Similar to the experiment of Example 30b, using pyrimidine-4-carbohydrazide, (3S)-6,7-dichloro-5-(2,6-difluorophenyl)-3-methyl-1,3-dihydro-1,4-benzodiazepine-2-thione was converted to the (-)-enantiopure (S)-labeled compound (71.6 mg, 31%) after chiral purification and obtained as a white solid. MS: 456.9 ([{ 35 Cl, 35 Cl}M+H] + ), ESI pos.

[0189] Example 53 (4S)-7,8-dichloro-6-(2,6-difluorophenyl)-4-methyl-1-(1-methylpyrazol-4-yl)-4H-[1,2,4]triazolo[4,3-a][1,4]benzodiazepine

Chemical formula

[0190] Similar to the experiment of Example 30b, using 1-methylpyrazole-4-carbohydrazide, (3S)-6,7-dichloro-5-(2,6-difluorophenyl)-3-methyl-1,3-dihydro-1,4-benzodiazepine-2-thione was converted to the (-)-enantiopure (S)-labeled compound (60.8 mg, 29%) after chiral purification and obtained as a white solid. MS: 459.1 ([{ 35 Cl, 35 Cl}M+H] +), ESI pos.

[0191] Example 56 (4S)-7,8-Dichloro-6-(2,6-difluorophenyl)-4-methyl-1-pyridazin-3-yl-4H-[1,2,4]triazolo[4,3-a][1,4]benzodiazepine [Chemical Structure]

[0192] Similar to the experiment of Example 30b, pyridazine-3-carbohydrazide was used to convert (3S)-6,7-dichloro-5-(2,6-difluorophenyl)-3-methyl-1,3-dihydro-1,4-benzodiazepine-2-thione into the (-)-enantiopure (S)-labeled compound (80.0 mg, 21%) after chiral purification and obtained as a white solid. MS: 457.0 ([{ 35 Cl, 35 Cl}M+H] + ), ESI pos.

[0193] Example 63 (4S)-7,8-Dichloro-6-(3-fluoro-2-pyridyl)-4-methyl-1-pyridazin-3-yl-4H-[1,2,4]triazolo[4,3-a][1,4]benzodiazepine [Chemical Structure]

[0194] a) (3S)-6,7-Dichloro-5-(3-fluoro-2-pyridyl)-3-methyl-1,3-dihydro-1,4-benzodiazepine-2-thione Similar to the experiment of Example 30a, (3S)-6,7-dichloro-5-(3-fluoro-2-pyridyl)-3-methyl-1,3-dihydro-1,4-benzodiazepine-2-one (Constituent O) was converted into the labeled compound (2.56 g, 67%) and obtained as a pale yellow solid. MS: 354.0 ([{ 35 Cl, 35 Cl}M+H] +), ESI pos.

[0195] b) (4S)-7,8-Dichloro-6-(3-fluoro-2-pyridyl)-4-methyl-1-pyridazin-3-yl-4H-[1,2,4]triazolo[4,3-a][1,4]benzodiazepine Similar to the experiment of Example 30b, (3S)-6,7-dichloro-5-(3-fluoro-2-pyridyl)-3-methyl-1,3-dihydro-1,4-benzodiazepine-2-thione was converted to the (+)-enantiopure (S)-labeled compound (8.8 mg, 2%) after chiral purification and obtained as a white solid. MS: 440.1 ([{ 35 Cl, 35 Cl}M+H] + ), ESI pos.

[0196] Example 64 (4S)-7,8-Dichloro-6-(2,6-difluorophenyl)-4-methyl-2,4-dihydro-[1,2,4]triazolo[4,3-a][1,4]benzodiazepin-1-one

Chem.

[0197] a) (3S)-6,7-Dichloro-5-(2,6-difluorophenyl)-3-methyl-1,3-dihydro-1,4-benzodiazepine-2-one hydrazone

[0198] (3S)-6,7-Dichloro-5-(2,6-difluorophenyl)-3-methyl-1,3-dihydro-1,4-benzodiazepine-2-thione (400 mg, 1.08 mmol) in tetrahydrofuran (8.7 mL) was added hydrazine monohydrate (109 mg, 104 μl, 2.15 mmol) at room temperature. The mixture was stirred under argon at room temperature for 2 h. The suspension was concentrated in vacuo to give a pale yellow solid which was treated with methyl tert-butyl ether (2 mL) and diluted with pentane (4 mL). The mixture was decanted to give a suspension which was stirred for 10 min. The solid was filtered, washed with pentane (2 × 3 mL) and dried in high vacuum to give the title compound (300 mg, 75%) as a yellow solid. MS: 369.1([{ 35 Cl, 35 Cl}M+H] + ), ESI pos.

[0199] b) (4S)-7,8-Dichloro-6-(2,6-difluorophenyl)-4-methyl-2,4-dihydro-[1,2,4]triazolo[4,3-a][1,4]benzodiazepin-1-one

[0200] (3S)-6,7-Dichloro-5-(2,6-difluorophenyl)-3-methyl-1,3-dihydro-1,4-benzodiazepine-2-one hydrazone (250 mg, 0.68 mmol) in tetrahydrofuran (6 mL) was added 1,1'-carbonyldiimidazole (132 mg, 0.81 mmol) at room temperature. The reaction mixture was stirred at 70 °C for 5 h and then concentrated in vacuo. The residue was purified by SFC (Chiralcel OJ-3, 0.05% diethylamine in methanol, 5 - 40%) to give the (-)-enantiopure (S)-title compound (141.2 mg, 53%) as a white solid. MS: 395.1({ 35 Cl, 35 Cl}[M+H] + ), ESI pos.

[0201] Example 66 (4S)-7,8-Dichloro-6-(3-fluoro-2-pyridyl)-4-methyl-1-pyrimidin-4-yl-4H-[1,2,4]triazolo[4,3-a][1,4]benzodiazepine

Chem.

[0202] Similar to the experiment of Example 30b, using pyrimidine-4-carbohydrazide, (3S)-6,7-dichloro-5-(3-fluoro-2-pyridyl)-3-methyl-1,3-dihydro-1,4-benzodiazepine-2-thione was converted to the (+)-enantiopure (S)-labeled compound (17 mg, 3%) after chiral purification and obtained as a white solid. MS: 440.1 ([{ 35 Cl, 35 Cl}M+H] + ), ESI pos.

[0203] Example 67 (4S)-7,8-Dichloro-6-(3-fluoro-2-pyridyl)-4-methyl-1-(1-methylpyrazol-4-yl)-4H-[1,2,4]triazolo[4,3-a][1,4]benzodiazepine

Chem.

[0204] Similar to the experiment of Example 30b, using 1-methylpyrazole-4-carbohydrazide, (3S)-6,7-dichloro-5-(3-fluoro-2-pyridyl)-3-methyl-1,3-dihydro-1,4-benzodiazepine-2-thione was converted to the (+)-enantiopure (S)-labeled compound (86 mg, 46%) after chiral purification and obtained as a white solid. MS: 442.2 ([{ 35 Cl, 35 Cl}M+H] + ), ESI pos.

[0205] Example 68 (4S)-7,8-Dichloro-1-cyclopropyl-6-(3-fluoro-2-pyridyl)-4-methyl-4H-[1,2,4]triazolo[4,3-a][1,4]benzodiazepine

Chem.

[0206] Similar to the experiment of Example 30b, using cyclopropanecarbohydrazide, (3S)-6,7-dichloro-5-(3-fluoro-2-pyridyl)-3-methyl-1,3-dihydro-1,4-benzodiazepine-2-thione was converted to the chiral-purified (+)-enantiopure (S)-labeled compound (123 mg, 21%) after chiral purification and obtained as a white solid. MS: 402.1 ([{ 35 Cl, 35 Cl}M+H] + ), ESI pos.

[0207] Example 73 (4S)-7-Bromo-8-chloro-6-(2,6-difluorophenyl)-4-methyl-1-(pyridazin-3-yl)-4H-benzo[f][1,2,4]triazolo[4,3-a][1,4]diazepine

Chem.

[0208] a) (3S)-6-Bromo-7-chloro-5-(2,6-difluorophenyl)-3-methyl-1,3-dihydro-1,4-benzodiazepine-2-thione Similar to the experiment of Example 30a, (3S)-6-bromo-7-chloro-5-(2,6-difluorophenyl)-3-methyl-1,3-dihydro-1,4-benzodiazepine-2-one (Component Q) was converted to the labeled compound (2.72 g, 77%) and obtained as a yellow powder. MS: 415.1 ([{ 79 Br, 35 Cl}M+H] + ), 417.0 ([{ 81 Br, 35 Cl or79 Br, 37 Cl}M+H] + ), ESI pos. b)(4S)-7-Bromo-8-chloro-6-(2,6-difluorophenyl)-4-methyl-1-pyridazin-3-yl-4H-[1,2,4]triazolo[4,3-a][1,4]benzodiazepine

[0209] Similar to the experiment of Example 30b, using pyridazine-3-carbohydrazide, (3S)-6-bromo-7-chloro-5-(2,6-difluorophenyl)-3-methyl-1,3-dihydro-1,4-benzodiazepine-2-thione was converted to the (-)-enantiopure (S)-labeled compound (24 mg, 20%) after chiral purification and obtained as a white solid. MS: 501.0([{ 79 Br, 35 Cl}M+H] + ), 503.1([{ 81 Br, 35 Cl or 79 Br, 37 Cl}M+H] + ), ESI pos.

[0210] Example 74 (4S)-7-Bromo-8-chloro-6-(2,6-difluorophenyl)-4-methyl-1-pyrimidin-4-yl-4H-[1,2,4]triazolo[4,3-a][1,4]benzodiazepine

Chemical Structure

[0211] Similar to the experiment of Example 30b, using pyrimidine-4-carbohydrazide, (3S)-6-bromo-7-chloro-5-(2,6-difluorophenyl)-3-methyl-1,3-dihydro-1,4-benzodiazepine-2-thione was converted to the (-)-enantiopure (S)-labeled compound (13 mg, 11%) after chiral purification and obtained as a white solid. MS: 501.1([{ 79 Br, 35 Cl}M+H] +), 503.0([{ 81 Br, 35 Cl or 79 Br, 37 Cl}M+H] + ), ESI pos.

[0212] Example 75 (4S)-8-Bromo-7-chloro-6-(2,6-difluorophenyl)-1,4-dimethyl-4H-[1,2,4]triazolo[4,3-a][1,4]benzodiazepine

Chem.

[0213] Similar to the experiment of Example 30b, using acetohydrazide, (3S)-7-bromo-6-chloro-5-(2,6-difluorophenyl)-3-methyl-1,3-dihydro-1,4-benzodiazepine-2-thione was converted to the (-)-enantiopure (S)-labeled compound (36 mg, 23%) after chiral purification and obtained as a white solid. MS: 437.0([{ 79 Br, 35 Cl}M+H] + ), 439.1([{ 81 Br, 35 Cl or 79 Br, 37 Cl}M+H] + ), ESI pos.

[0214] Example 76 (4S)-7-Bromo-8-chloro-6-(2,6-difluorophenyl)-4-methyl-1-(1-methylpyrazol-4-yl)-4H-[1,2,4]triazolo[4,3-a][1,4]benzodiazepine

Chem.

[0215] Similar to the experiment of Example 30b, using 1-methylpyrazole-4-carbohydrazide, (3S)-6-bromo-7-chloro-5-(2,6-difluorophenyl)-3-methyl-1,3-dihydro-1,4-benzodiazepine-2-thione was converted to the (-)-enantiopure (S)-labeled compound (13 mg, 11%) after chiral purification, and a white solid was obtained. MS: 503.1 ([{ 79 Br, 35 Cl}M+H] + ), 505.1 ([{ 81 Br, 35 Cl or 79 Br, 37 Cl}M+H] + ), ESI pos.

[0216] Example 80 (4S)-7-bromo-8-chloro-6-(2,6-difluorophenyl)-1,4-dimethyl-4H-[1,2,4]triazolo[4,3-a][1,4]benzodiazepine

Chemical Structure

[0217] Similar to the experiment of Example 30b, using acetohydrazide, (3S)-6-bromo-7-chloro-5-(2,6-difluorophenyl)-3-methyl-1,3-dihydro-1,4-benzodiazepine-2-thione was converted to the (-)-enantiopure (S)-labeled compound (13 mg, 11%) after chiral purification, and obtained as a white solid. MS: 436.9 ([{ 79 Br, 35 Cl}M+H] + ), 438.9 ([{ 81 Br, 35 Cl or 79 Br, 37 Cl}M+H] + ), ESI pos.

[0218] Example 81 (4S)-7-Bromo-8-chloro-6-(2,6-difluorophenyl)-4-methyl-1-(1-methylpyrazol-3-yl)-4H-[1,2,4]triazolo[4,3-a][1,4]benzodiazepine

Chem.

[0219] Similar to the experiment of Example 30b, using 1-methylpyrazole-3-carbohydrazide, (3S)-6-bromo-7-chloro-5-(2,6-difluorophenyl)-3-methyl-1,3-dihydro-1,4-benzodiazepine-2-thione was converted to the (-)-enantiopure (S)-labeled compound (38 mg, 26%) after chiral purification and obtained as a white solid. MS: 503.0 ([{ 79 Br, 35 Cl}M+H] + ), 505.1 ([{ 81 Br, 35 Cl or 79 Br, 37 Cl}M+H] + ), ESI pos.

[0220] Example 82 (4S)-7,8-Dichloro-6-(2,6-difluorophenyl)-1,4-dimethyl-4H-[1,2,4]triazolo[4,3-a][1,4]benzodiazepine

Chem.

[0221] Similar to the experiment of Example 30b, using acetohydrazide, (3S)-6,7-dichloro-5-(2,6-difluorophenyl)-3-methyl-1,3-dihydro-1,4-benzodiazepine-2-thione was converted to the (-)-enantiopure (S)-labeled compound (62.3 mg, 19%) after chiral purification and obtained as a white solid. MS: 393.0 ([{ 35 Cl, 35 Cl}M+H] + ), ESI pos.

[0222] Example 83 (4S)-8-Bromo-7-chloro-1-cyclopropyl-6-(3-fluoro-2-pyridyl)-4-methyl-4H-[1,2,4]triazolo[4,3-a][1,4]benzodiazepine

Chemical Structure

[0223] a) (3S)-7-Bromo-6-chloro-5-(3-fluoro-2-pyridyl)-3-methyl-1,3-dihydro-1,4-benzodiazepine-2-thione Similar to the experiment of Example 30a, (3S)-7-bromo-6-chloro-5-(3-fluoro-2-pyridyl)-3-methyl-1,3-dihydro-1,4-benzodiazepine-2-one (constituent R) was converted to the title compound (720 mg, 46%) and obtained as a yellow solid. MS: 398.0 ([{ 79 Br, 35 Cl}M+H] + ), 400.1 ([{ 81 Br, 35 Cl or 79 Br, 37 Cl}M+H] + ), ESI pos.

[0224] b) (4S)-8-Bromo-7-chloro-1-cyclopropyl-6-(3-fluoro-2-pyridyl)-4-methyl-4H-[1,2,4]triazolo[4,3-a][1,4]benzodiazepine

[0225] Similar to the experiment of Example 30b, using cyclopropanecarbohydrazide, (3S)-7-bromo-6-chloro-5-(3-fluoro-2-pyridyl)-3-methyl-1,3-dihydro-1,4-benzodiazepine-2-thione was converted to the chiral-purified (+)-enantiopure (S)-title compound (38 mg, 11%) and obtained as a pale yellow solid. MS: 446.1 ([{ 79 Br, 35 Cl}M+H] +), 448.0 ([{ 81 Br, 35 Cl or 79 Br, 37 Cl}M+H] + ), ESI pos.

[0226] Example 84 (4S)-8-Bromo-7-chloro-6-(3-fluoro-2-pyridyl)-4-methyl-4H-[1,2,4]triazolo[4,3-a][1,4]benzodiazepine

Chemical Structure

[0227] Similar to the experiment of Example 30b, using formohydrazide, (3S)-7-bromo-6-chloro-5-(3-fluoro-2-pyridyl)-3-methyl-1,3-dihydro-1,4-benzodiazepine-2-thione was converted to the chiral-purified (+)-enantiopure (S)-labeled compound (38 mg, 12%) and obtained as a pale yellow solid. MS: 406.0 ([{ 79 Br, 35 Cl}M+H] + ), 408.1 ([{ 81 Br, 35 Cl or 79 Br, 37 Cl}M+H] + ), ESI pos.

[0228] Example 85 (4S)-8-Bromo-7-chloro-6-(3-fluoro-2-pyridyl)-1,4-dimethyl-4H-[1,2,4]triazolo[4,3-a][1,4]benzodiazepine

Chemical Structure

[0229] Similar to the experiment of Example 30b, using acetohydrazide, (3S)-7-bromo-6-chloro-5-(3-fluoro-2-pyridyl)-3-methyl-1,3-dihydro-1,4-benzodiazepine-2-thione was converted to the (+)-enantiopure (S)-labeled compound (31 mg, 8%) after chiral purification and obtained as a white solid. MS: 420.0 ([{ 79 Br, 35 Cl}M+H] + ), 422.0 ([{ 81 Br, 35 Cl or 79 Br, 37 Cl}M+H] + ), ESI pos.

[0230] Example 86 (4S)-7,8-dichloro-6-(2,6-difluorophenyl)-4-methyl-1-(6-methylpyridazin-3-yl)-4H-[1,2,4]triazolo[4,3-a][1,4]benzodiazepine

Chemical Structure

[0231] a) 6-methylpyridazine-3-carbohydrazide

[0232] A solution of ethyl 6-methylpyridazine-3-carboxylate (3.71 g, 22.3 mmol) in methanol (40 mL) was heated to 60 °C. After 10 minutes, hydrazine monohydrate (1.62 mL, 33.5 mmol) was carefully added and the reaction mixture was cooled to room temperature. After adding diethyl ether (60 mL), the reaction mixture was cooled to 0 °C. After 2 hours, the resulting suspension was filtered through a sintered funnel. The recovered solid was washed with diethyl ether and dried under high vacuum to obtain the title compound (1.4 g, 41%), obtained as an off-white powder. MS: 153.1 ([M+H] + ), ESI pos.

[0233] b) (4S)-7,8-Dichloro-6-(2,6-difluorophenyl)-4-methyl-1-(6-methylpyridazin-3-yl)-4H-[1,2,4]triazolo[4,3-a][1,4]benzodiazepine

[0234] Similar to the experiment of Example 30b, using 6-methylpyridazine-3-carbohydrazide, (3S)-6,7-dichloro-5-(2,6-difluorophenyl)-3-methyl-1,3-dihydro-1,4-benzodiazepine-2-thione was converted to the (-)-enantiopure (S)-labeled compound (52.6 mg, 44%) after chiral purification and obtained as a white solid. MS: 471.1([{ 35 Cl, 35 Cl}M+H] + ), ESI pos.

[0235] Example 87 5-[(4S)-7,8-Dichloro-6-(2,6-difluorophenyl)-4-methyl-4H-[1,2,4]triazolo[4,3-a][1,4]benzodiazepin-1-yl]-3-methyl-isoxazole

Chemical Structure

[0236] a) 3-Methyl-1,2-didehydroisoxazole-5-carbohydrazide Similar to the experiment of Example 86a, methyl 3-methyl-1,2-didehydroisoxazole-5-carboxylate was converted to the labeled compound (800 mg, 74%) and obtained as a white solid. MS: 142.1([M+H] + ), ESI pos.

[0237] b) 5-[(4S)-7,8-Dichloro-6-(2,6-difluorophenyl)-4-methyl-4H-[1,2,4]triazolo[4,3-a][1,4]benzodiazepin-1-yl]-3-methyl-isoxazole

[0238] Similar to the experiment of Example 30b, using 3-methyl-1,2-didehydroisoxazole-5-carbohydrazide, (3S)-6,7-dichloro-5-(2,6-difluorophenyl)-3-methyl-1,3-dihydro-1,4-benzodiazepine-2-thione was converted to the (-)-enantiopure (S)-labeled compound (54.1 mg, 31%) after chiral purification and obtained as a white solid. MS: 460.1 ([{ 35 Cl, 35 Cl}M+H] + ), ESI pos.

[0239] Example 88 (4S)-7-Bromo-8-chloro-6-(2,6-difluorophenyl)-4-methyl-1-(6-methylpyridazin-3-yl)-4H-[1,2,4]triazolo[4,3-a][1,4]benzodiazepine

Chemical Structure

[0240] Similar to the experiment of Example 30b, using 6-methylpyridazine-3-carbohydrazide, (3S)-6-bromo-7-chloro-5-(2,6-difluorophenyl)-3-methyl-1,3-dihydro-1,4-benzodiazepine-2-thione was converted to the (-)-enantiopure (S)-labeled compound (30 mg, 24%) after chiral purification and obtained as a white solid. MS: 515.0 ([{ 79 Br, 35 Cl}M+H] + ), 517.0 ([{ 81 Br, 35 Cl or 79 Br, 37 Cl}M+H] + ), ESI pos.

[0241] Example 89 (4S)-7-Bromo-8-chloro-6-(2,6-difluorophenyl)-4-methyl-2,4-dihydro-[1,2,4]triazolo[4,3-a][1,4]benzodiazepine-1-one [Chemical]

[0242] a) (3S)-6-Bromo-7-chloro-5-(2,6-difluorophenyl)-3-methyl-1,3-dihydro-1,4-benzodiazepin-2-one hydrazone Similar to the experiment of Example 64a, (3S)-6-bromo-7-chloro-5-(2,6-difluorophenyl)-3-methyl-1,3-dihydro-1,4-benzodiazepine-2-thione was converted to the labeled compound (40 mg, 20%) and obtained as a powder. MS: 413.0 ([{ 79 Br, 35 Cl}M+H] + ), 415.0 ([{ 81 Br, 35 Cl or 79 Br, 37 Cl}M+H] + ), ESI pos.

[0243] b) (4S)-7-Bromo-8-chloro-6-(2,6-difluorophenyl)-4-methyl-2,4-dihydro-[1,2,4]triazolo[4,3-a][1,4]benzodiazepin-1-one

[0244] Similar to the experiment of Example 64b, (3S)-6-bromo-7-chloro-5-(2,6-difluorophenyl)-3-methyl-1,3-dihydro-1,4-benzodiazepine-2-one hydrazone was converted to the (-)-enantiopure (S)-labeled compound (12 mg, 28%) after chiral purification and obtained as a white solid. MS: 439.0 ([{ 79 Br, 35 Cl}M+H] + ), 441.1 ([{ 81 Br, 35 Cl or 79 Br, 37 Cl}M+H] + ), ESI pos.

[0245] Example 92 (4S)-7-chloro-6-(2,6-difluorophenyl)-1,4-dimethyl-8-(trifluoromethyl)-4H-[1,2,4]triazolo[4,3-a][1,4]benzodiazepine

Chemical formula

[0246] a) (3S)-6-chloro-5-(2,6-difluorophenyl)-3-methyl-7-(trifluoromethyl)-1,3-dihydro-1,4-benzodiazepin-2-one

[0247] (3S)-6-chloro-5-(2,6-difluorophenyl)-3-methyl-7-iodo-1,3-dihydro-1,4-benzodiazepin-2-one (constituent S, 500 mg, 1.12 mmol), copper iodide (426 mg, 2.24 mmol), hexamethylphosphoramide (2.5 mL, 1.12 mmol) and a solution of methyl 2,2-difluoro-2-fluorosulfonyl-acetate (645 mg, 3.36 mmol) in N,N-dimethylformamide (5 mL) were stirred at 70 °C for 16 h. Methyl 2,2-difluoro-2-fluorosulfonyl-acetate (430 mg, 2.24 mmol) and copper iodide (213 mg, 1.12 mmol) were added and the reaction mixture was stirred at 70 °C for a further 4 h. The mixture was diluted with ethyl acetate (150 mL), washed with saturated aqueous ammonium chloride solution (80 mL), and the organic layer was filtered through a sintered funnel. The filtrate was washed with water (50 mL) and brine (50 mL), dried over sodium sulfate and concentrated under reduced pressure. The crude product was purified by flash column chromatography (silica, petroleum ether / ethyl acetate, 20:1 to 1:1) to give the title compound (550 mg, 127%), obtained as a dark red oil. MS: 389.0 ([M+H] + ), ESI pos.

[0248] b) (3S)-6-chloro-5-(2,6-difluorophenyl)-3-methyl-7-(trifluoromethyl)-1,3-dihydro-1,4-benzodiazepine-2-thione Similar to the experiment of Example 30a, (3S)-6-chloro-5-(2,6-difluorophenyl)-3-methyl-7-(trifluoromethyl)-1,3-dihydro-1,4-benzodiazepin-2-one was converted to the labeled compound (400 mg, 77%) and obtained as a yellow solid. MS: 405.0 ([M+H] + ), ESI pos.

[0249] c) (4S)-7-chloro-6-(2,6-difluorophenyl)-1,4-dimethyl-8-(trifluoromethyl)-4H-[1,2,4]triazolo[4,3-a][1,4]benzodiazepine Similar to the experiment of Example 30b, using acetohydrazide, (3S)-6-chloro-5-(2,6-difluorophenyl)-3-methyl-7-(trifluoromethyl)-1,3-dihydro-1,4-benzodiazepine-2-thione was converted to the (-)-enantiopure (S)-labeled compound (63.5 mg, 32%) after chiral purification and obtained as a white solid. MS: 427.1 ([M+H] + ), ESI pos.

[0250] Example 93 (4S)-7-chloro-8-(1,1-difluoroethyl)-6-(2,6-difluorophenyl)-1,4-dimethyl-4H-[1,2,4]triazolo[4,3-a][1,4]benzodiazepine

Chemical formula

[0251] a) (4S)-7-chloro-6-(2,6-difluorophenyl)-8-(1-ethoxyvinyl)-1,4-dimethyl-4H-[1,2,4]triazolo[4,3-a][1,4]benzodiazepine

[0252] (4S)-8-Bromo-7-chloro-6-(2,6-difluorophenyl)-1,4-dimethyl-4H-[1,2,4]triazolo[4,3-a][1,4]benzodiazepine (Example 75, 210.0 mg, 0.480 mmol) and tributyl(1-ethoxyvinyl)tin (346.57 mg, 0.960 mmol) in N,N-dimethylformamide (2.1 mL) were suspended, and tetrakis(triphenylphosphine)palladium(0) (56.42 mg, 0.050 mmol) was added. The reaction mixture was stirred at 80 °C for 1 hour. The mixture was diluted with dichloromethane (2 × 20 mL), washed with water (20 mL) and brine (20 mL), dried over sodium sulfate, and concentrated under reduced pressure. The residue was purified by flash column chromatography (silica, petroleum ether / ethyl acetate 10:1, dichloromethane / methanol 80:1) to give the title compound (300 mg, 98%) as a colorless oil. MS: 429.1 ([M+H] + ), ESI pos.

[0253] b) 1-[(4S)-7-Chloro-6-(2,6-difluorophenyl)-1,4-dimethyl-4H-[1,2,4]triazolo[4,3-a][1,4]benzodiazepin-8-yl]ethanone

[0254] (4S)-7-Chloro-6-(2,6-difluorophenyl)-8-(1-ethoxyvinyl)-1,4-dimethyl-4H-[1,2,4]triazolo[4,3-a][1,4]benzodiazepine (280 mg, 0.650 mmol) in 1,4-dioxane (7 mL) was added with aqueous hydrochloric acid solution (2.0 m, 1.62 mL, 3.23 mmol). The mixture was stirred at room temperature for 0.5 hour and then diluted with dichloromethane (100 mL). The organic layer was washed with water (3 × 10 mL), aqueous sodium hydrogen carbonate solution (3 × 50 mL) and brine (50 mL), then dried over sodium sulfate and concentrated under reduced pressure. The residue was purified by preparative HPLC (BostonPrime C18, 0.1% trifluoroacetic acid / acetonitrile in water) and lyophilized to give the title compound (220 mg, 84%), which was obtained as a pale yellow solid. MS: 401.1 ([M+H]+ ), ESI pos.

[0255] c) (4S)-7-chloro-8-(1,1-difluoroethyl)-6-(2,6-difluorophenyl)-1,4-dimethyl-4H-[1,2,4]triazolo[4,3-a][1,4]benzodiazepine

[0256] To a solution of 1-[(4S)-7-chloro-6-(2,6-difluorophenyl)-1,4-dimethyl-4H-[1,2,4]triazolo[4,3-a][1,4]benzodiazepin-8-yl]ethanone (100 mg, 0.250 mmol) in dichloroethane (2 mL) was added diethylaminosulfur trifluoride (120.6 mg, 0.750 mmol) at 0 °C. The mixture was stirred at room temperature for 24 h and then poured into saturated aqueous sodium hydrogen carbonate (20 mL). The mixture was extracted with dichloromethane (2 × 50 mL). The combined organic extracts were washed with brine (30 mL), dried over sodium sulfate, and concentrated in vacuo. The residue was purified by preparative HPLC (Phenomenex Synergi C18, 0.1% trifluoroacetic acid / water / acetonitrile) to give the (-)-enantiopure (S)-labeled compound (20.1 mg, 18%) as a white solid. MS: 423.0 ([M+H] + ), ESI pos.

[0257] Example 94 (4S)-7,8-dichloro-6-(2,6-difluorophenyl)-4-methyl-1-(6-methylpyrimidin-4-yl)-4H-[1,2,4]triazolo[4,3-a][1,4]benzodiazepine

Chemical formula

[0258] Similar to the experiment of Example 30b, using 6-methylpyrimidine-4-carbohydrazide, (3S)-6,7-dichloro-5-(2,6-difluorophenyl)-3-methyl-1,3-dihydro-1,4-benzodiazepine-2-thione was converted to the (-)-enantiopure (S)-labeled compound (70.0 mg, 35%) after chiral purification and obtained as a white solid. MS: 471.2 ([{ 35 Cl, 35 Cl}M+H] + ), ESI pos.

[0259] Example 95 (4S)-7,8-Dichloro-6-(2,6-difluorophenyl)-4-methyl-1-(2-methylpyrimidin-4-yl)-4H-[1,2,4]triazolo[4,3-a][1,4]benzodiazepine

Chemical formula

[0260] Similar to the experiment of Example 30b, using 2-methylpyrimidine-4-carbohydrazide, (3S)-6,7-dichloro-5-(2,6-difluorophenyl)-3-methyl-1,3-dihydro-1,4-benzodiazepine-2-thione was converted to the (-)-enantiopure (S)-labeled compound (12.0 mg, 20%) after chiral purification and obtained as a white solid. MS: 471.2 ([{ 35 Cl, 35 Cl}M+H] + ), ESI pos.

[0261] Example 96 (4S)-7,8-Dichloro-6-(2,6-difluorophenyl)-1-(2,6-dimethylpyrimidin-4-yl)-4-methyl-4H-[1,2,4]triazolo[4,3-a][1,4]benzodiazepine

Chemical formula

[0262] Similar to the experiment of Example 30b, using 2,6-dimethylpyrimidine-4-carbohydrazide, (3S)-6,7-dichloro-5-(2,6-difluorophenyl)-3-methyl-1,3-dihydro-1,4-benzodiazepine-2-thione was converted to the (-)-enantiopure (S)-labeled compound (66.8 mg, 44%) after chiral purification and obtained as a white solid. MS: 485.1([{ 35 Cl, 35 Cl}M+H] + ), ESI pos.

[0263] Example 97 (4S)-7-chloro-6-(2,6-difluorophenyl)-1,4,8-trimethyl-4H-[1,2,4]triazolo[4,3-a][1,4]benzodiazepine

Chemical formula

[0264] (4S)-8-bromo-7-chloro-6-(2,6-difluorophenyl)-1,4-dimethyl-4H-[1,2,4]triazolo[4,3-a][1,4]benzodiazepine (300 mg, 0.690 mmol), tetrakis(triphenylphosphine)palladium(0) (79.2 mg, 0.070 mmol), trimethylaluminum (2.0 M in toluene, 0.51 mL, 1.03 mmol) in N,N-dimethylformamide (6 mL) was heated to 70 °C. After 16 hours, the mixture was diluted with dichloromethane (30 mL). The organic layer was washed with water (20 mL), brine (30 mL), dried over sodium sulfate and concentrated under reduced pressure. The residue was purified by preparative HPLC (BostonPrime, 0.1% trifluoroacetic acid in water / acetonitrile) and lyophilized to give the (-)-enantiopure (S)-labeled compound (27.3 mg, 58%), which was obtained as a white solid. MS: 373.2([M+H] + ), ESI pos.

[0265] Example 98 (4S)-8-Bromo-7-chloro-6-(2,6-difluorophenyl)-4-ethyl-1-methyl-4H-[1,2,4]triazolo[4,3-a][1,4]benzodiazepine

Chem.

[0266] a) (3S)-7-Bromo-6-chloro-5-(2,6-difluorophenyl)-3-ethyl-1,3-dihydro-1,4-benzodiazepine-2-thione Similar to the experiment of Example 30a, (3S)-7-bromo-6-chloro-5-(2,6-difluorophenyl)-3-ethyl-1,3-dihydro-1,4-benzodiazepine-2-one (Component T) was converted to the title compound (600 mg, 92%) and obtained as a yellow solid. MS: 429.0 ([{ 79 Br, 35 Cl}M+H] + ), 431.1 ([{ 81 Br, 35 Cl or 79 Br, 37 Cl}M+H] + ), ESI pos.

[0267] b) (3S)-7-Bromo-6-chloro-5-(2,6-difluorophenyl)-3-ethyl-1,3-dihydro-1,4-benzodiazepine-2-one hydrazone

[0268] (3S)-7-Bromo-6-chloro-5-(2,6-difluorophenyl)-3-ethyl-1,3-dihydro-1,4-benzodiazepine-2-thione (500 mg, 1.16 mmol) and hydrazine hydrate (117 mg, 2.33 mmol) in tetrahydrofuran (5 mL) were cooled to 15 °C and stirred for 1 hour. The mixture was concentrated under reduced pressure to obtain the title compound (450 mg, 90%) as a pale green foam. MS: 427.2 ([{ 79 Br, 35 Cl}M+H] + ), 429.3 ([{ 81 Br, 35 Cl or79 Br, 37 Cl}M+H] + ), ESI pos.

[0269] c)(4S)-8-bromo-7-chloro-6-(2,6-difluorophenyl)-4-ethyl-1-methyl-4H-[1,2,4]triazolo[4,3-a][1,4]benzodiazepine

[0270] (3S)-7-bromo-6-chloro-5-(2,6-difluorophenyl)-3-ethyl-1,3-dihydro-1,4-benzodiazepin-2-one hydrazone (450 mg, 1.05 mmol) and triethyl orthoacetate (854 mg, 5.26 mmol) in toluene (5 mL) were heated to 120 °C. After 1 hour, the reaction mixture was concentrated under reduced pressure. The residue was purified directly by flash column chromatography (dichloromethane / methanol 20:1) and subsequently by chiral purification to give the (-)-enantiopure (S)-labeled compound (400 mg, 82%) as a pale yellow foam. MS: 451.0 ([{ 79 Br, 35 Cl}M+H] + ), 453.0 ([{ 81 Br, 35 Cl or 79 Br, 37 Cl}M+H] + ), ESI pos.

[0271] Example 99 (4S)-7-chloro-8-(difluoromethyl)-6-(2,6-difluorophenyl)-1,4-dimethyl-4H-[1,2,4]triazolo[4,3-a][1,4]benzodiazepine

Chemical Structure

[0272] a)(4S)-7-chloro-6-(2,6-difluorophenyl)-1,4-dimethyl-8-vinyl-4H-[1,2,4]triazolo[4,3-a][1,4]benzodiazepine

[0273] (4S)-8-Bromo-7-chloro-6-(2,6-difluorophenyl)-1,4-dimethyl-4H-[1,2,4]triazolo[4,3-a][1,4]benzodiazepine (3.0 g, 6.85 mmol) in ethanol (70 mL) was added with potassium vinyltrifluoroborate (1.85 g, 1.37 mmol), triethylamine (2.08 g, 20.56 mmol), [1,1'-bis(diphenylphosphino)ferrocene]dichloropalladium(II) (0.5 g, 0.690 mmol). The reaction mixture was heated to 80 °C. After 16 h, water (100 mL) was added and the reaction mixture was extracted with ethyl acetate (2 × 100 mL). The combined organic extracts were washed with brine (100 mL), dried over sodium sulfate, filtered, and concentrated under reduced pressure. The residue was purified by flash column chromatography (petroleum ether / ethyl acetate / ethanol 20:3:1~8:3:1) to give the title compound (2.8 g, 88%), which was obtained as a brown solid. MS: 385.1([M+H] + ), ESI pos.

[0274] b) (4S)-7-Chloro-6-(2,6-difluorophenyl)-1,4-dimethyl-4H-[1,2,4]triazolo[4,3-a][1,4]benzodiazepine-8-carbaldehyde

[0275] (4S)-7-chloro-6-(2,6-difluorophenyl)-1,4-dimethyl-8-vinyl-4H-[1,2,4]triazolo[4,3-a][1,4]benzodiazepine (2.8 g, 7.28 mmol) in a solution of acetone (56 mL) and water (14 mL) was added osmium tetroxide (184.99 mg, 0.730 mmol). After stirring at room temperature for 10 minutes, sodium periodate (3.1 g, 14.5 mmol) was added and the mixture was stirred at room temperature for an additional 1 hour. The reaction mixture was diluted with water (10 mL) and ethyl acetate (100 mL). The organic layer was washed with brine (2 × 50 mL), dried over sodium sulfate, filtered, and concentrated under reduced pressure. The residue was purified by flash column chromatography (silica, dichloromethane / methanol 200:1 to 60:1) to give the title compound (1.8 g, 47%) as a yellow solid. MS: 387.1 ([M+H] + ), ESI pos.

[0276] c) (4S)-7-chloro-8-(difluoromethyl)-6-(2,6-difluorophenyl)-1,4-dimethyl-4H-[1,2,4]triazolo[4,3-a][1,4]benzodiazepine

[0277] (4S)-7-chloro-6-(2,6-difluorophenyl)-1,4-dimethyl-4H-[1,2,4]triazolo[4,3-a][1,4]benzodiazepine-8-carbaldehyde (1.8 g, 4.65 mmol) in dichloroethane (37.0 mL) was added with diethylaminosulfur trifluoride (2.25 g, 14.0 mmol) at 0 °C. After the addition, the reaction mixture was warmed to room temperature and stirred for an additional 2 hours. The reaction mixture was quenched by the addition of saturated aqueous sodium bicarbonate solution (10 mL) and then extracted with dichloromethane (2 × 30 mL). The combined organic extracts were washed with brine (2 × 50 mL), filtered, and concentrated under reduced pressure. The residue was purified by preparative HPLC (BostonPrimeC18, 0.1% trifluoroacetic acid / water / acetonitrile) and lyophilized to give the (-)-enantiopure (S)-labeled compound (359 mg, 50%) as a white solid. MS: 409.1 ([M+H] + ), ESI pos.

[0278] Example 100 (4R)-8-bromo-7-chloro-6-(2,6-difluorophenyl)-4-(methoxymethyl)-1-methyl-4H-[1,2,4]triazolo[4,3-a][1,4]benzodiazepine

Chemical formula

[0279] a) (3S)-7-bromo-6-chloro-5-(2,6-difluorophenyl)-3-(methoxymethyl)-1,3-dihydro-1,4-benzodiazepine-2-thione

[0280] In the same experiment as in Example 30a, (3S)-7-bromo-6-chloro-5-(2,6-difluorophenyl)-3-(methoxymethyl)-1,3-dihydro-1,4-benzodiazepine-2-one (Constituent U) was converted to the labeled compound (1.2 g, 76%) and obtained as a yellow solid. MS: 445.0 ([{ 79 Br, 35 Cl}M+H] +), 447.1 ([{ 81 Br, 35 Cl or 79 Br, 37 Cl}M+H] + ), ESI pos.

[0281] b) (3R)-7-Bromo-6-chloro-5-(2,6-difluorophenyl)-3-(methoxymethyl)-1,3-dihydro-1,4-benzodiazepin-2-one hydrazone Similar to the experiment of Example 98b, (3S)-7-bromo-6-chloro-5-(2,6-difluorophenyl)-3-(methoxymethyl)-1,3-dihydro-1,4-benzodiazepine-2-thione was converted to the labeled compound (1.1 g, 96%) and obtained as a yellow solid. MS: 443.1 ([{ 79 Br, 35 Cl}M+H] + ), 445.1 ([{ 81 Br, 35 Cl or 79 Br, 37 Cl}M+H] + ), ESI pos.

[0282] c) (4R)-8-Bromo-7-chloro-6-(2,6-difluorophenyl)-4-(methoxymethyl)-1-methyl-4H-[1,2,4]triazolo[4,3-a][1,4]benzodiazepine

[0283] Similar to the experiment of Example 98c, (3R)-7-bromo-6-chloro-5-(2,6-difluorophenyl)-3-(methoxymethyl)-1,3-dihydro-1,4-benzodiazepin-2-one hydrazone was converted to the (-)-enantiopure (R)-labeled compound (68 mg, 6%) after chiral purification and obtained as a white solid. MS: 467.2 ([{ 79 Br, 35 Cl}M+H] + ), 469.1 ([{ 81 Br, 35 Cl or 79 Br, 37 Cl}M+H] + ), ESI pos.

[0284] Example 101 [(4R)-8-Bromo-7-chloro-6-(2,6-difluorophenyl)-1-methyl-4H-[1,2,4]triazolo[4,3-a][1,4]benzodiazepin-4-yl]methanol

Chemical formula

[0285] (4R)-8-Bromo-7-chloro-6-(2,6-difluorophenyl)-4-(methoxymethyl)-1-methyl-4H-[1,2,4]triazolo[4,3-a][1,4]benzodiazepine (230.0 mg, 0.490 mmol) and sodium iodide (147.43 mg, 0.980 mmol) in dichloromethane (3 mL) were stirred to form a suspension. A solution of boron tribromide (308.0 mg, 1.23 mmol) in dichloromethane (0.5 mL) was added thereto at -30 °C. The reaction mixture was warmed to room temperature, stirred for an additional 1 hour, and then concentrated under reduced pressure. The residue was purified by preparative HPLC (Phenomenex luna, 0.1% trifluoroacetic acid / water / acetonitrile), and then by SFC (Daicel Chiralpak AD-H, 0.1% ammonia / methanol) to obtain the (-)-enantiopure (R)-labeled compound (71 mg, 47%) as a white solid. MS: 453.2 ([{ 79 Br, 35 Cl}M+H] + ), 455.2 ([{ 81 Br, 35 Cl or 79 Br, 37 Cl}M+H] + ), ESI pos.

[0286] Example 102 (4S)-7-Chloro-6-(2,6-difluorophenyl)-8-iodo-1,4-dimethyl-4H-[1,2,4]triazolo[4,3-a][1,4]benzodiazepine

Chemical formula

[0287] (4S)-8-Bromo-7-chloro-6-(2,6-difluorophenyl)-1,4-dimethyl-4H-[1,2,4]triazolo[4,3-a][1,4]benzodiazepine (100 mg, 0.228 mmol), rac-trans-N1,N2-dimethylcyclohexane-1,2-diamine (35.3 μL, 0.228 mmol), sodium iodide (342 mg, 2.28 mmol) and copper(I) iodide (21.8 mg, 114 μmol) were added 1,4-dioxane (10 mL) under argon. The resulting suspension was heated at 115 °C for 6 days. Additional amounts of sodium iodide (685 mg, 4.57 mmol) and copper(I) iodide (218 mg, 1.14 mmol) were added and the reaction mixture was stirred for a further 2 days. The mixture was diluted with ethyl acetate and the organic layer was washed twice with aqueous ammonia and brine, dried over sodium sulfate and concentrated in vacuo. The crude material was purified by flash column chromatography (silica, 0-15% methanol in dichloromethane), followed by chiral HPLC (Chiracel OD; eluent: 20% in heptane (0.1 mol ammonium acetate in ethanol)) to give the (-)-enantiopure (S)-labeled compound (41.2 mg, 37%) as a white solid. MS: 485.0 ([M+H] + ), ESI pos.

[0288] Example 103 [(4S)-7-Chloro-6-(2,6-difluorophenyl)-4-methyl-8-(trifluoromethyl)-4H-[1,2,4]triazolo[4,3-a][1,4]benzodiazepin-1-yl]methanol

Chemical formula

[0289] a) [(4S)-7-Chloro-6-(2,6-difluorophenyl)-4-methyl-8-(trifluoromethyl)-4H-[1,2,4]triazolo[4,3-a][1,4]benzodiazepin-1-yl]methoxy triisopropylsilane Similar to the experiment of Example 30b, using 2-triisopropylsilyloxyacetohydrazide, (3S)-6-chloro-5-(2,6-difluorophenyl)-3-methyl-7-(trifluoromethyl)-1,3-dihydro-1,4-benzodiazepine-2-thione was converted to the labeled compound (300 mg, 33%) and obtained as a pale yellow oil. MS: 599.2 ([M+H] + ), ESI pos.

[0290] b) [(4S)-7-chloro-6-(2,6-difluorophenyl)-4-methyl-8-(trifluoromethyl)-4H-[1,2,4]triazolo[4,3-a][1,4]benzodiazepin-1-yl]methanol

[0291] To a solution of [(4S)-7-chloro-6-(2,6-difluorophenyl)-4-methyl-8-(trifluoromethyl)-4H-[1,2,4]triazolo[4,3-a][1,4]benzodiazepin-1-yl]methoxy-triisopropylsilane (200.0 mg, 0.270 mmol) in tetrahydrofuran (4 mL) was slowly added TBAF (1.0 M in tetrahydrofuran, 0.81 mL, 0.810 mmol). The mixture was stirred at room temperature for 1 hour and then concentrated under reduced pressure. The residue was purified by flash column chromatography (dichloromethane / methanol 50:1 to 20:1), followed by preparative HPLC (UniSil3-100C18Uitra, 0.225% trifluoroacetic acid / water / acetonitrile) and SFC separation (Daicel Chiralcel OJ, 0.1% ammonia in ethanol) to obtain the (-)-enantiopure (S)-labeled compound (13.6 mg, 10%) as a white solid. MS: 443.0 ([M+H] + ), ESI pos.

[0292] Example 105 (4R)-7-chloro-6-(2,6-difluorophenyl)-4-(methoxymethyl)-1-methyl-8-(trifluoromethyl)-4H-[1,2,4]triazolo[4,3-a][1,4]benzodiazepine [Chemical formula]

[0293] To a vial dried with a magnetic stir bar and a dryer equipped with a Teflon septum, (4R)-8-bromo-7-chloro-6-(2,6-difluorophenyl)-4-(methoxymethyl)-1-methyl-4H-[1,2,4]triazolo[4,3-a][1,4]benzodiazepine (10 mg, 21.4 μmol), sodium carbonate (9.06 mg, 85.5 μmol), CuBr2·2LiBr (4.28 μmol), Mes-Umemoto reagent (22 mg, 42.8 μmol), Ir[dFMeppy]2-(4,4’-dCF3bpy)PF6 (56 μg, 0.0535 μmol) and (Me3Si)3SiOH (8.5 mg, 32.1 μmol) were added. Then, the vial was evacuated and refilled with nitrogen, degassed, and then degassed acetone (0.2 mL) was added by syringe. The reaction mixture was stirred at room temperature for 16 h under irradiation with a blue LED (Kessil lamp 40W, 420 nm). The vial was opened and the reaction mixture was filtered directly through a Celite plug. The filter cake was rinsed with ethyl acetate (2.0 mL) and the filtrate was concentrated under reduced pressure. The residue was dissolved in ethyl acetate (20 mL), washed with water and brine (3×15 mL), dried over sodium sulfate, filtered, and concentrated under reduced pressure. The crude material was purified by preparative HPLC (Gemini NX 5Y, 0.1% formic acid / water / acetonitrile) to give the (-)-enantiopure (R)-labeled compound (3 mg, 30%) as a white solid. MS: 457.2 ([M+H] + ), ESI pos.

[0294] Example 106 [(4R)-7-chloro-6-(2,6-difluorophenyl)-1-methyl-8-(trifluoromethyl)-4H-[1,2,4]triazolo[4,3-a][1,4]benzodiazepin-4-yl]methanol [Chemical formula]

[0295] Similar to the experiment of Example 105, [(4R)-8-bromo-7-chloro-6-(2,6-difluorophenyl)-1-methyl-4H-[1,2,4]triazolo[4,3-a][1,4]benzodiazepin-4-yl]methanol was converted to the (-)-enantiopure (R)-labeled compound (9 mg, 31%) and obtained as a white lyophilized powder. MS: 443.1 ([M+H] + ), ESI pos.

[0296] Example 107 (4S)-7-chloro-6-(2,6-difluorophenyl)-4-methyl-8-(trifluoromethyl)-4H-[1,2,4]triazolo[4,3-a][1,4]benzodiazepine

Chemical formula

[0297] Similar to the experiment of Example 105, (4S)-8-bromo-7-chloro-6-(2,6-difluorophenyl)-4-methyl-4H-[1,2,4]triazolo[4,3-a][1,4]benzodiazepine was converted to the (-)-enantiopure (S)-labeled compound (1 mg, 10%) and obtained as a white solid. MS: 413.2 ([M+H] + ), ESI pos.

[0298] Example 108 (4S)-7-chloro-6-(2,6-difluorophenyl)-1-ethyl-4-methyl-8-(trifluoromethyl)-4H-[1,2,4]triazolo[4,3-a][1,4]benzodiazepine

Chemical formula

[0299] Similar to the experiment of Example 30b, using propanohydrazide, (3S)-6-chloro-5-(2,6-difluorophenyl)-3-methyl-7-(trifluoromethyl)-1,3-dihydro-1,4-benzodiazepine-2-thione was converted to the (-)-enantiopure (S)-labeled compound (16.4 mg, 13%) after chiral purification and obtained as a white solid. MS: 444.1([M+H] + ), ESI pos.

[0300] Example 109 (4S)-7-chloro-6-(2,6-difluorophenyl)-8-ethyl-1,4-dimethyl-4H-[1,2,4]triazolo[4,3-a][1,4]benzodiazepine

Chemical formula

[0301] (4S)-8-bromo-7-chloro-6-(2,6-difluorophenyl)-1,4-dimethyl-4H-[1,2,4]triazolo[4,3-a][1,4]benzodiazepine (120 mg, 0.274 mmol) and [1,1'-bis(diphenylphosphino)ferrocene]dichloropalladium(II)·CH2Cl2 (4.5 mg, 5.5 μmol) in anhydrous tetrahydrofuran (1.2 mL) were added dropwise with diethylzinc (1.0 M in heptane, 0.823 mL, 0.823 mmol) at 0 °C. After warming the reaction mixture to room temperature, it was heated at 55 °C for 15 hours. The reaction mixture was poured into water and extracted twice with ethyl acetate. The combined organic layers were washed with water and brine, dried over sodium sulfate, filtered, and concentrated under reduced pressure. The crude material was purified by preparative HPLC (Gemini NX, 0.1% triethylamine / methanol in water), followed by chiral HPLC (ReprosilChiral NR, 0.01 M ammonium acetate in ethanol, 30%) to obtain the (-)-enantiopure (S)-labeled compound (31.7 mg, 30%) as a white powder. MS: 387.3([M+H] + ), ESI pos.

[0302] (Reference compound) RE-A 8-Bromo-6-(2,6-difluorophenyl)-1-methyl-4H-[1,2,4]triazolo[4,3-a][1,4]benzodiazepine

Chemical formula

[0303] a) N-[2-(2,6-Difluorobenzoyl)phenyl]acetamide Similar to the experiment of Component Ab, 2-methyl-3,1-benzoxazin-4-one (CAS No. 525-76-8) was converted to the title compound (40 g, 80%) and obtained as a pale yellow solid. MS: 276.2 ([M+H] + ), ESI pos.

[0304] b) (2-Aminophenyl)-(2,6-difluorophenyl)methanone Similar to the experiment of Component Ac, N-[2-(2,6-difluorobenzoyl)phenyl]acetamide was converted to the title compound (19.5 g, 75%) and obtained as a yellow solid. MS: 234.1 ([M+H] + ), ESI pos.

[0305] c) (2-Amino-5-bromo-phenyl)-(2,6-difluorophenyl)methanone

[0306] A solution of (2-aminophenyl)-(2,6-difluorophenyl)methanone (5.00 g, 21.4 mmol) in dichloromethane (50 mL) was added N-bromosuccinimide (4.02 g, 22.51 mmol) portionwise at -15 °C. The reaction mixture was stirred at -15 °C for 1 h until the starting material was completely consumed (judged by LCMS analysis). The mixture was concentrated under reduced pressure and the resulting residue was purified by preparative HPLC (Shim-pack C18, 0.225% trifluoroacetic acid / water / acetonitrile). The combined fractions were diluted with saturated aqueous sodium hydrogen carbonate solution and extracted with ethyl acetate (3 × 200 mL). The organic phase was washed with brine (2 × 100 mL), dried (Na2SO4), concentrated under reduced pressure to give the title compound (4.44 g, 66%) as a yellow solid. MS: 311.9 ([{ 79 Br}M+H] + ), 314.0 ([{ 81 Br}M+H] + ), ESI pos.

[0307] d) 7-Bromo-5-(2,6-difluorophenyl)-1,3-dihydro-1,4-benzodiazepin-2-one Following the experiment of Component Ae, (2-amino-5-bromo-phenyl)-(2,6-difluorophenyl)methanone was converted to the title compound (300 mg, 13%) and obtained as a yellow solid. MS: 351.0 ([{ 79 Br}M+H] + ), 353.0 ([{ 81 Br}M+H] + ), ESI pos.

[0308] e) 7-Bromo-5-(2,6-difluorophenyl)-1,3-dihydro-1,4-benzodiazepine-2-thione

[0309] Following the experiment of Example 30a, 7-bromo-5-(2,6-difluorophenyl)-1,3-dihydro-1,4-benzodiazepin-2-one was converted to the title compound (310 mg, 92%) and obtained as a yellow solid. The crude product was used directly in the next step without further characterization.

[0310] f) 8-Bromo-6-(2,6-difluorophenyl)-1-methyl-4H-[1,2,4]triazolo[4,3-a][1,4]benzodiazepine Similar to the experiment of Example 30b, using acetohydrazide, 7-bromo-5-(2,6-difluorophenyl)-1,3-dihydro-1,4-benzodiazepine-2-thione was converted to the labeled compound (5.1 mg, 4%) and obtained as a white solid. MS: 389.0 ([{ 79 Br}M+H] + ), 391.0 ([{ 81 Br}M+H] + ), ESI pos.

[0311] RE-B 6-(2,6-Difluorophenyl)-1-methyl-8-(trifluoromethyl)-4H-[1,2,4]triazolo[4,3-a][1,4]benzodiazepine

Chemical Structure

[0312] a) (2-Amino-5-iodo-phenyl)-(2,6-difluorophenyl)methanone

[0313] (2-Aminophenyl)-(2,6-difluorophenyl)methanone (1.6 g, 6.86 mmol) in DMF (15 mL) solution, N-iodosuccinimide (1.62 g, 7.2 mmol) was added little by little. After the reaction mixture was stirred at 20 °C for 16 hours, it was diluted with water (20 mL). The mixture was extracted with ethyl acetate (3 × 20 mL), then the combined organic extracts were washed with brine (2 × 10 mL), dried (Na2SO4), and concentrated under reduced pressure. The residue was purified by flash column chromatography (petroleum ether / ethyl acetate, 10:1 to 5:1) to obtain the labeled compound (2.0 g, 81%) as a yellow solid. MS: 360.0 ([M+H] + ), ESI pos.

[0314] b) 5-(2,6-Difluorophenyl)-7-iodo-1,3-dihydro-1,4-benzodiazepin-2-one Similar to the experiment of Component Ae, (2-amino-5-iodo-phenyl)-(2,6-difluorophenyl)methanone was converted to the labeled compound (650 mg, 12%) and obtained as a yellow solid. MS: 399.0 ([M+H] + ), ESI pos.

[0315] c) 5-(2,6-Difluorophenyl)-7-iodo-1,3-dihydro-1,4-benzodiazepine-2-thione Similar to the experiment of Example 30a, 5-(2,6-difluorophenyl)-7-iodo-1,3-dihydro-1,4-benzodiazepin-2-one was converted to the labeled compound (200 mg, 82%) and obtained as a yellow solid. MS: 414.9 ([M+H] + ), ESI pos.

[0316] d) 5-(2,6-Difluorophenyl)-7-iodo-1,3-dihydro-1,4-benzodiazepin-2-one hydrazone Similar to the experiment of Example 64a, 5-(2,6-difluorophenyl)-7-iodo-1,3-dihydro-1,4-benzodiazepine-2-thione was converted to the labeled compound (220 mg, 98%) and obtained as a yellow solid. MS: 413.0 ([M+H] + ), ESI pos.

[0317] e) 6-(2,6-Difluorophenyl)-8-iodo-1-methyl-4H-[1,2,4]triazolo[4,3-a][1,4]benzodiazepine Similar to the experiment of Example 98c, 5-(2,6-difluorophenyl)-7-iodo-1,3-dihydro-1,4-benzodiazepin-2-one hydrazone was converted to the labeled compound (150 mg, 64%) and obtained as a yellow solid. MS: 437.0 ([M+H] + ), ESI pos.

[0318] f) 6-(2,6-Difluorophenyl)-1-methyl-8-(trifluoromethyl)-4H-[1,2,4]triazolo[4,3-a][1,4]benzodiazepine In the same experiment as Example 92a, 6-(2,6-difluorophenyl)-8-iodo-1-methyl-4H-[1,2,4]triazolo[4,3-a][1,4]benzodiazepine was converted to the title compound (5 mg, 6%) and obtained as a white solid. MS: 379.0 ([M+H] + ), ESI pos.

[0319] RE-C 6-(2,6-Difluorophenyl)-1-methyl-4H-[1,2,4]triazolo[4,3-a][1,4]benzodiazepine

Chemical formula

[0320] To a stirred solution of 8-bromo-6-(2,6-difluorophenyl)-1-methyl-4H-[1,2,4]triazolo[4,3-a][1,4]benzodiazepine (30 mg, 0.08 mmol) in methanol (0.5 mL) was added 10 wt% Pd / C (2.5 mg, 2.4 μmol). The resulting black suspension was purged by evacuation and then filled three times with a hydrogen stream (balloon). The mixture was stirred at room temperature for 16 h under a hydrogen atmosphere and then filtered through a pad of dicalite. The filter cake was rinsed with methanol and the filtrate was concentrated under reduced pressure. The residue was purified by preparative HPLC (Phenomenex Gemini-NX C18, 0.1% trifluoroacetic acid / water / acetonitrile) followed by preparative TLC (silica, dichloromethane / methanol, 20:1) to give the title compound (5 mg, 20%) as a white solid. MS: 276.2 ([M+H] + ), ESI pos.

[0321] RE-D (4S)-7-chloro-6-(2,6-difluorophenyl)-1,4-dimethyl-4H-[1,2,4]triazolo[4,3-a][1,4]benzodiazepine [Chemical formula]

[0322] Similar to the experiment of reference compound RE-C, (4S)-8-bromo-7-chloro-6-(2,6-difluorophenyl)-1,4-dimethyl-4H-[1,2,4]triazolo[4,3-a][1,4]benzodiazepine was converted to the title compound (2 mg, 15%) and obtained as a white solid. MS: 359.1 ([M+H] + ), ESI pos.

Claims

1. Formula (I): 【Chemistry 1】 (In the formula, R １ teeth, i) H; ii) C １－６ -alkyl, iii) C １－６ -alkoxy, iv) C １－６ -Alkoxy-C １－６ -alkyl, v) hydroxy, vi) Hydroxy-C １－６ -alkyl, vii) optionally R ７ , R ８ and R ９ C substituted with ３－８ -cycloalkyl, viii) Amino-C １－６ -alkyl, ix) optionally R ７ , R ８ and R ９ Heteroaryl substituted with, x) optionally R ７ , R ８ and R ９ Heterocycloalkyl substituted with Selected from: R ２ teeth, i) C １－６ -alkyl, ii) hydroxy, iii) Hydroxy-C １－６ -alkyl, and iv) C １－６ -Alkoxy-C １－６ -Alkyl Selected from: R ３ teeth, i) Cl, and ii) F Selected from: X is i) CR ６ , and ii) N Selected from: R ６ teeth, i) H; ii) Cl, and iii) F Selected from: R ４ teeth, i) Br, and ii) Cl Selected from: R ５ teeth, i) C １－６ -alkyl, ii) C １－６ -alkoxy, iii) halogens, iv) halo-C １－６ -alkyl, v) cyano, and vi) C ３－８ -Cycloalkyl Selected from: R ７ , R ８ and R ９ teeth, i) C １－６ -alkyl, and ii) C １－６ -Alkoxy (independently selected from or a pharma- ceutically acceptable salt thereof.

2. R １ but, i) H; ii) C １－６ -alkyl, iii) C １－６ -alkoxy, iv) C １－６ -Alkoxy-C １－６ -alkyl, v) hydroxy, vi) Hydroxy-C １－６ -alkyl, vii) optionally R ７ , R ８ and R ９ C substituted with ３－８ -cycloalkyl, viii) Amino-C １－６ -alkyl, ix) optionally R ７ , R ８ and R ９ pyrazolyl substituted with x) optionally R ７ , R ８ and R ９ pyridinyl substituted with xi) optionally R ７ , R ８ and R ９ pyrimidinyl substituted with xii) optionally R ７ , R ８ and R ９ pyridazinyl substituted with xiii) optionally R ７ , R ８ and R ９ Isoxazolyl substituted with Selected from: R ２ but, i) C １－６ -alkyl, ii) hydroxy, iii) Hydroxy-C １－６ -alkyl, and iv) C １－６ -Alkoxy-C １－６ -Alkyl Selected from: R ３ but, i) Cl, and ii) F Selected from: X, i) CR ６ , and ii) N Selected from: R ６ but, i) H; ii) Cl, and iii) F Selected from: R ４ but, i) Br, and ii) Cl Selected from: R ５ but, i) C １－６ -alkyl, ii) C １－６ -alkoxy, iii) halogens, iv) halo-C １－６ -alkyl, v) C ３－８ -Cycloalkyl Selected from: R ７ , R ８ and R ９ but, i) C １－６ -alkyl, and ii) C １－６ -Alkoxy are independently selected from 2. A compound of formula (I) according to claim 1, or a pharma- ceutically acceptable salt thereof.

3. R １ but, i) H; ii) C １－６ -alkyl, iii) hydroxy, iv) Hydroxy-C １－６ -alkyl, v) optionally R ７ , R ８ and R ９ C substituted with ３－８ -cycloalkyl, vi) optionally R ７ , R ８ and R ９ pyrazolyl substituted with vii) optionally R ７ , R ８ and R ９ pyrimidinyl substituted with viii) optionally R ７ , R ８ and R ９ pyridazinyl substituted with, ix) optionally R ７ , R ８ and R ９ Isoxazolyl substituted with Selected from: R ２ but, i) C １－６ -alkyl, ii) hydroxy, iii) Hydroxy-C １－６ -alkyl, and iv) C １－６ -Alkoxy-C １－６ -Alkyl Selected from: R ３ is F, X, i) CR ６ , and ii) N Selected from: R ６ but, i) H, and ii) F Selected from: R ４ but, i) Br, and ii) Cl Selected from: R ５ but, i) C １－６ -alkyl, ii) a halogen, and iii) Halo-C １－６ -Alkyl Selected from: R ７ , R ８ and R ９ But, C １－６ independently selected from alkyl, 3. A compound of formula (I) according to claim 1 or 2, or a pharma- ceutically acceptable salt thereof.

4. R １ But, C １－６ is alkyl, R ２ But, C １－６ is alkyl, R ３ is F, X is CR ６ and R ６ is F, R ４ is Cl, R ５ But, Hello-C １－６ - alkyl, 4. The compound according to any one of claims 1 to 3, or a pharma- ceutically acceptable salt thereof.

5. (4S)-8-bromo-7-chloro-6-(2-fluorophenyl)-4-methyl-4H-[1,2,4]triazolo[4,3-a][1,4]benzodiazepine; 8-bromo-7-chloro-6-(2,6-difluorophenyl)-4H-[1,2,4]triazolo[4,3-a][1,4]benzodiazepin-4-ol; (4S)-8-bromo-7-chloro-6-(2-fluorophenyl)-4-methyl-1-pyridazin-3-yl-4H-[1,2,4]triazolo[4,3-a][1,4]benzodiazepine; 8-bromo-7-chloro-6-(2-fluorophenyl)-1-pyridazin-3-yl-4H-[1,2,4]triazolo[4,3-a][1,4]benzodiazepin-4-ol; (4S)-8-bromo-7-chloro-6-(2,6-difluorophenyl)-4-methyl-4H-[1,2,4]triazolo[4,3-a][1,4]benzodiazepine; (4S)-7,8-dichloro-6-(2,6-difluorophenyl)-4-methyl-1-pyrimidin-4-yl-4H-[1,2,4]triazolo[4,3-a][1,4]benzodiazepine; (4S)-7,8-dichloro-6-(2,6-difluorophenyl)-4-methyl-1-(1-methylpyrazol-4-yl)-4H-[1,2,4]triazolo[4,3-a][1,4]benzodiazepine; (4S)-7,8-dichloro-6-(2,6-difluorophenyl)-4-methyl-1-pyridazin-3-yl-4H-[1,2,4]triazolo[4,3-a][1,4]benzodiazepine; (4S)-7,8-dichloro-6-(3-fluoro-2-pyridyl)-4-methyl-1-pyridazin-3-yl-4H-[1,2,4]triazolo[4,3-a][1,4]benzodiazepine; (4S)-7,8-dichloro-6-(2,6-difluorophenyl)-4-methyl-2,4-dihydro-[1,2,4]triazolo[4,3-a][1,4]benzodiazepin-1-one; (4S)-7,8-dichloro-6-(3-fluoro-2-pyridyl)-4-methyl-1-pyrimidin-4-yl-4H-[1,2,4]triazolo[4,3-a][1,4]benzodiazepine; (4S)-7,8-dichloro-6-(3-fluoro-2-pyridyl)-4-methyl-1-(1-methylpyrazol-4-yl)-4H-[1,2,4]triazolo[4,3-a][1,4]benzodiazepine; (4S)-7,8-dichloro-1-cyclopropyl-6-(3-fluoro-2-pyridyl)-4-methyl-4H-[1,2,4]triazolo[4,3-a][1,4]benzodiazepine; (4S)-7-bromo-8-chloro-6-(2,6-difluorophenyl)-4-methyl-1-pyridazin-3-yl-4H-[1,2,4]triazolo[4,3-a][1,4]benzodiazepine; (4S)-7-bromo-8-chloro-6-(2,6-difluorophenyl)-4-methyl-1-pyrimidin-4-yl-4H-[1,2,4]triazolo[4,3-a][1,4]benzodiazepine; (4S)-8-bromo-7-chloro-6-(2,6-difluorophenyl)-1,4-dimethyl-4H-[1,2,4]triazolo[4,3-a][1,4]benzodiazepine; (4S)-7-bromo-8-chloro-6-(2,6-difluorophenyl)-4-methyl-1-(1-methylpyrazol-4-yl)-4H-[1,2,4]triazolo[4,3-a][1,4]benzodiazepine; (4S)-7-bromo-8-chloro-6-(2,6-difluorophenyl)-1,4-dimethyl-4H-[1,2,4]triazolo[4,3-a][1,4]benzodiazepine; (4S)-7-bromo-8-chloro-6-(2,6-difluorophenyl)-4-methyl-1-(1-methylpyrazol-3-yl)-4H-[1,2,4]triazolo[4,3-a][1,4]benzodiazepine; (4S)-7,8-dichloro-6-(2,6-difluorophenyl)-1,4-dimethyl-4H-[1,2,4]triazolo[4,3-a][1,4]benzodiazepine; (4S)-8-bromo-7-chloro-1-cyclopropyl-6-(3-fluoro-2-pyridyl)-4-methyl-4H-[1,2,4]triazolo[4,3-a][1,4]benzodiazepine; (4S)-8-bromo-7-chloro-6-(3-fluoro-2-pyridyl)-4-methyl-4H-[1,2,4]triazolo[4,3-a][1,4]benzodiazepine; (4S)-8-bromo-7-chloro-6-(3-fluoro-2-pyridyl)-1,4-dimethyl-4H-[1,2,4]triazolo[4,3-a][1,4]benzodiazepine; (4S)-7,8-dichloro-6-(2,6-difluorophenyl)-4-methyl-1-(6-methylpyridazin-3-yl)-4H-[1,2,4]triazolo[4,3-a][1,4]benzodiazepine; 5-[(4S)-7,8-dichloro-6-(2,6-difluorophenyl)-4-methyl-4H-[1,2,4]triazolo[4,3-a][1,4]benzodiazepin-1-yl]-3-methyl-isoxazole; (4S)-7-bromo-8-chloro-6-(2,6-difluorophenyl)-4-methyl-1-(6-methylpyridazin-3-yl)-4H-[1,2,4]triazolo[4,3-a][1,4]benzodiazepine; (4S)-7-Bromo-8-chloro-6-(2,6-difluorophenyl)-4-methyl-2,4-dihydro-[1,2,4]triazolo[4,3-a][1,4]benzodiazepin-1-one; (4S)-7-chloro-6-(2,6-difluorophenyl)-1,4-dimethyl-8-(trifluoromethyl)-4H-[1,2,4]triazolo[4,3-a][1,4]benzodiazepine; (4S)-7-chloro-8-(1,1-difluoroethyl)-6-(2,6-difluorophenyl)-1,4-dimethyl-4H-[1,2,4]triazolo[4,3-a][1,4]benzodiazepine; (4S)-7,8-dichloro-6-(2,6-difluorophenyl)-4-methyl-1-(6-methylpyrimidin-4-yl)-4H-[1,2,4]triazolo[4,3-a][1,4]benzodiazepine; (4S)-7,8-dichloro-6-(2,6-difluorophenyl)-4-methyl-1-(2-methylpyrimidin-4-yl)-4H-[1,2,4]triazolo[4,3-a][1,4]benzodiazepine; (4S)-7,8-dichloro-6-(2,6-difluorophenyl)-1-(2,6-dimethylpyrimidin-4-yl)-4-methyl-4H-[1,2,4]triazolo[4,3-a][1,4]benzodiazepine; (4S)-7-chloro-6-(2,6-difluorophenyl)-1,4,8-trimethyl-4H-[1,2,4]triazolo[4,3-a][1,4]benzodiazepine; (4S)-8-bromo-7-chloro-6-(2,6-difluorophenyl)-4-ethyl-1-methyl-4H-[1,2,4]triazolo[4,3-a][1,4]benzodiazepine; (4S)-7-chloro-8-(difluoromethyl)-6-(2,6-difluorophenyl)-1,4-dimethyl-4H-[1,2,4]triazolo[4,3-a][1,4]benzodiazepine; (4R)-8-bromo-7-chloro-6-(2,6-difluorophenyl)-4-(methoxymethyl)-1-methyl-4H-[1,2,4]triazolo[4,3-a][1,4]benzodiazepine; [(4R)-8-bromo-7-chloro-6-(2,6-difluorophenyl)-1-methyl-4H-[1,2,4]triazolo[4,3-a][1,4]benzodiazepin-4-yl]methanol; (4S)-7-chloro-6-(2,6-difluorophenyl)-8-iodo-1,4-dimethyl-4H-[1,2,4]triazolo[4,3-a][1,4]benzodiazepine; [(4S)-7-chloro-6-(2,6-difluorophenyl)-4-methyl-8-(trifluoromethyl)-4H-[1,2,4]triazolo[4,3-a][1,4]benzodiazepin-1-yl]methanol; (4R)-7-chloro-6-(2,6-difluorophenyl)-4-(methoxymethyl)-1-methyl-8-(trifluoromethyl)-4H-[1,2,4]triazolo[4,3-a][1,4]benzodiazepine; [(4R)-7-chloro-6-(2,6-difluorophenyl)-1-methyl-8-(trifluoromethyl)-4H-[1,2,4]triazolo[4,3-a][1,4]benzodiazepin-4-yl]methanol; (4S)-7-chloro-6-(2,6-difluorophenyl)-4-methyl-8-(trifluoromethyl)-4H-[1,2,4]triazolo[4,3-a][1,4]benzodiazepine; (4S)-7-chloro-6-(2,6-difluorophenyl)-1-ethyl-4-methyl-8-(trifluoromethyl)-4H-[1,2,4]triazolo[4,3-a][1,4]benzodiazepine; (4S)-7-chloro-6-(2,6-difluorophenyl)-8-ethyl-1,4-dimethyl-4H-[1,2,4]triazolo[4,3-a][1,4]benzodiazepine; 5. The compound according to any one of claims 1 to 4, or a pharma- ceutically acceptable salt thereof, selected from:

6. The compound is (4S)-7-chloro-6-(2,6-difluorophenyl)-1,4-dimethyl-8-(trifluoromethyl)-4H-[1,2,4]triazolo[4,3-a][1,4]benzodiazepine, or a pharma- ceutically acceptable salt thereof.

7. A compound of formula (III) and a compound of formula (IV) １ , R ２ , R ３ , R ４ and R ５ is as defined in any one of claims 1 to 4) with a compound of formula (I) as defined in any one of claims 1 to 4. 【Chemistry 2】

8. A compound according to any one of claims 1 to 6 for use as a therapeutically active substance.

9. A pharmaceutical composition comprising a compound according to any one of claims 1 to 6 and a therapeutically inert carrier.

10. A compound according to any one of claims 1 to 6 for use in the treatment or prevention of autism spectrum disorders, Rett syndrome, post-traumatic stress disorder and fragile X disorder.

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