RU2476220C2 - (2s,3r-)-n-(2-((3-pyridinyl)methyl)-1-azabicyclo[2,2,2]oct-3-yl)benzofurane-2-carboxamide, new salt forms and methods for preparing them - Google Patents

Abstract

FIELD: medicine, pharmaceutics.

SUBSTANCE: present invention refers to organic chemistry and medicine, and concerns salts of (2S,3R-)-N-(2-((3-pyridinyl)methyl)-1-azabicyclo[2.2.2]oct-3-yl)benzofurane-2-carboxamide, substantially free from (2S,3S)-N-(2-((3-pyridinyl)methyl)-1-azabicyclo[2.2.2]oct-3-yl)benzofurane-2-carboxamide, (2R,3S)-N-(2-((3-pyridinyl)methyl)-1-azabicyclo[2.2.2]oct-3-yl)benzofurane-2-carboxamide and (2R,3R)-N-(2-((3-pyridinyl)methyl)-1-azabicyclo[2.2.2]oct-3-yl)benzofurane-2-carboxamide.

EFFECT: preparing the compositions applicable for treating the conditions and disorders associated with central and vegetative nervous system dysfunction.

5 cl, 19 dwg, 7 tbl, 18 ex

Description

CROSS RELATIONSHIP WITH PRIORITY APPLICATIONS

The present invention claims priority based on provisional patent applications US No. 60/971654, filed September 12, 2007; 60/953610, filed August 2, 2007; 60/953613, filed August 2, 2007, and 60/953614, filed August 2, 2007, each of which is incorporated into this invention by reference in its entirety.

FIELD OF THE INVENTION

The present invention relates to (2S, 3R) -N- (2 - ((3-pyridinyl) methyl) -1-azabicyclo [2.2.2] oct-3-yl) benzofuran-2-carboxamide, its new salt forms, methods their preparation, new intermediate compounds and methods of treating a large number of conditions and disorders, including those associated with dysfunction of the central and autonomic nervous system.

BACKGROUND OF THE INVENTION

It has been shown that neuronal nicotinic receptors (NNRs), characteristic of the central nervous system (CNS), exist as several subtypes, the most common of which are the subtypes α4β2 and α7. See, for example, Schmitt, Current Med. Chem. 7: 749 (2000), incorporated herein by reference. It has been suggested that ligands that interact with the α7 NNR subtype can be used in the treatment of a variety of disease states and disorders. A basic understanding of the α7 subtype of the neuronal nicotinic receptor is disclosed in Mazurov et al., Curr. Med. Chem. 13: 1567-1584 (2006) and the references cited therein, incorporated herein by reference. Well-known analogues of such conditions and disorders are impaired consciousness, schizophrenia, inflammation, angiogenesis, neurotic pain and fibromyalgia.

A post-mortem examination of the brain tissue of patients with schizophrenia revealed a reduced number of hippocampal NNRs. There is also an improved psychological effect in smokers with schizophrenia compared to non-smokers. Nicotine improves sensory deficiency in animals and schizophrenics. Blockade of the α7 NNR subtype induces a selection deficiency similar to that observed in schizophrenia. See, for example, Leonard et al., Schizophrenia Bulletin 22 (3): 431 (1996), incorporated herein by reference. Biochemical, molecular, and genetic studies of sensory processing in patients with P50 hearing impaired potential selection deficits suggest that the α7 NNR subtype may function as an inhibitory neural pathway. See, for example, Freedman et al., Biological Psychiatry 38 (1): 22 (1995), incorporated herein by reference.

It was later suggested that α7 NNRs are mediators of angiogenesis, as described in Heeschen et al., J. Clin. Invest., 100: 527 (2002), incorporated by reference. These studies have shown that inhibition of the α7 subtype reduces inflammatory angiogenesis. Α7 NNR receptors have also been proposed as targets for controlled neurogenesis and tumor growth (Utsugisawa et al., Molecular Brain Research 106 (1-2): 88 (2002) and US Patent Application 2002/0016371, each of which is incorporated into this invention by links). Finally, the role of the α7 subtype in cognitive ability (Levin and Rezvani, Current Drug Targets: CNS and Neurological Disorders 1 (4): 423 (2002)), neuroprotection (O'Neill et al., Current Drug Targets: CNS and Neurological Disorders 1 (4): 399 (2002) and Jeyarasasingam et al., Neuroscience 109 (2): 275 (2002)) and neuropathic pain (Xiao et al., Proc. Nat. Acad. Sci. (US) 99 ( 12): 8360 (2002)), each citation is incorporated herein by reference.

Various compounds have been reported to interact with α7 NNR receptors and, on this basis, they have been proposed as a treatment. See, for example, PCT WO 99/62505, PCT WO 99/03859, PCT WO 97/30998, PCT WO 01/36417, PCT WO 02/15662, PCT WO 02/16355, PCT WO 02/16356, PCT WO 02 / 16357, PCT WO 02/16358, PCT WO 02/17358, Stevens et al., Psychopharm. 136: 320 (1998), Dolle et al., J. Labelled Comp. Radiopharm. 44: 785 (2001) and Macor et al., Bioorg. Med. Chem. Lett. 11: 319 (2001) and the references cited therein, these references are incorporated into this description by reference to the previous indications of α7 NNR and the proposed treatment. Among these compounds, a common structural motif is a substituted tertiary bicyclic amine (e.g., quinuclidine). It has also been reported that similar substituted quinuclidine compounds bind to muscarinic receptors. See, for example, US Pat. Nos. 5,712,270, Sabb, and PCT WO 02/00652 and WO 02/051841, each of which is incorporated herein by reference for such compounds.

A limitation of certain nicotinic compounds is that they are associated with various undesirable side effects, for example, stimulation of muscle and ganglion receptors. This leads to the continuing need for compounds, compositions and methods for preventing or treating various conditions or disorders, such as CNS disorders, including alleviating the symptoms of these disorders, where the compounds exhibit a pharmacological effect characteristic of nicotinic compounds along with a beneficial effect, namely, the functioning of the central nervous system but without significant related side effects. There is a need for compounds, compositions and methods that affect the function of the central nervous system without significantly affecting those subtypes of nicotinic receptors that are capable of causing unwanted side effects, such as marked activity at the cardiovascular sites and skeletal muscle sites. The present invention provides such compounds, compositions and methods.

SUMMARY OF THE INVENTION

In one aspect, the present invention relates to (2S, 3R) -N- (2 - ((3-pyridinyl) methyl) -1-azabicyclo [2.2.2] oct-3-yl) benzofuran-2-carboxamide or a pharmaceutically acceptable thereof salt. Another aspect relates to (2S, 3R) -N- (2 - ((3-pyridinyl) methyl) -1-azabicyclo [2.2.2] oct-3-yl) benzofuran-2-carboxamide essentially in pure form or to its pharmaceutically acceptable salt. A further aspect relates to (2S, 3R) -N- (2 - ((3-pyridinyl) methyl) -1-azabicyclo [2.2.2] oct-3-yl) benzofuran-2-carboxamide substantially free of (2S , 3S), (2R, 3S) or (2R, 3R) isomers, or a pharmaceutically acceptable salt thereof.

Additionally, another aspect relates to stereochemically enriched (2S, 3R) -N- (2 - ((3-pyridinyl) methyl) -1-azabicyclo [2.2.2] oct-3-yl) benzofuran-2-carboxamide or its pharmaceutically acceptable salt. In one embodiment, the enantiomeric and / or diastereomeric excess is 90% or more. In one embodiment, the enantiomeric and / or diastereomeric excess is 95% or more. In one embodiment, the enantiomeric and / or diastereomeric excess is 98% or more. In one embodiment, the enantiomeric and / or diastereomeric excess is 99% or more. In one embodiment, the enantiomeric and / or diastereomeric excess is 99.5% or more.

Another aspect of the present invention relates to a salt of (2S, 3R) -N- (2 - ((3-pyridinyl) methyl) -1-azabicyclo [2.2.2] oct-3-yl) benzofuran-2-carboxamide with an acid, where the acid is selected from hydrochloric acid, sulfuric acid, phosphoric acid, maleic acid, p-toluenesulfonic acid, galactaric (mucous) acid, D-mandelic acid, D-tartaric acid, methanesulfonic acid, R- and S-10-camphorsulfonic acid, ketoglutaric acid acid or hippuric acid. In one embodiment, the stoichiometric ratio of (2S, 3R) -N- (2 - ((3-pyridinyl) methyl) -1-azabicyclo [2.2.2] oct-3-yl) benzofuran-2-carboxamide to acid is 2: 1, 1: 1 or 1: 2. In one embodiment, the stoichiometric ratio is 1: 1. One embodiment of the present invention is (2S, 3R) -N- (2 - ((3-pyridinyl) methyl) -1-azabicyclo [2.2.2] oct-3-yl) benzofuran-2-carboxamide hydrochloride or a hydrate thereof or a solvate, including partial hydrates or solvates. A further embodiment is (2S, 3R) -N- (2 - ((3-pyridinyl) methyl) -1-azabicyclo [2.2.2] oct-3-yl) benzofuran-2-carboxamide monohydrochloride or a hydrate or solvate thereof including partial hydrates or solvates.

The present invention also relates to scalable syntheses of (2S, 3R) -N- (2 - ((3-pyridinyl) methyl) -1-azabicyclo [2.2.2] oct-3-yl) benzofuran-2-carboxamide and new intermediates .

The scope of the present invention includes all combinations of aspects, embodiments, and advantages described herein.

BRIEF DESCRIPTION OF THE FIGURES

1A1-1A4 illustrates the responses of rat α7 receptors expressed in mammalian GH4C1 cells to (2S, 3R) -N- (2 - ((3-pyridinyl) methyl) -1-azabicyclo [2.2.2] oct- 3-yl) benzofuran-2-carboxamide; racemate, i.e. a mixture of (2S, 3R), (2R, 3S), (2R, 3R) and (2S, 3S); individual stereoisomers and acetylcholine (ACh).

FIG. 1B shows a comparison of the functional responses of rat α7 receptors expressed in mammalian GH4C1 cells with (2S, 3R) -N- (2 - ((3-pyridinyl) methyl) -1-azabicyclo [2.2.2] oct- 3-yl) benzofuran-2-carboxamide; racemate, i.e. a mixture of (2S, 3R), (2R, 3S), (2R, 3R) and (2S, 3S); and individual stereoisomers within the effective range of plasma concentrations.

FIG. 2A illustrates the responses of human α7 receptors expressed in Xenopus oocytes to (2S, 3R) -N- (2 - ((3-pyridinyl) methyl) -1-azabicyclo [2.2.2] oct-3-yl ) benzofuran-2-carboxamide.

2B illustrates control responses of human α7 receptors after administration of the compound at the indicated concentrations. The data were normalized to the resulting charge of control responses to 300 μM Ach obtained 5 minutes before the experimental agonist-induced responses. Each point represents the mean ± statistical error (SEM) of the normalized response for at least 4 oocytes.

Figure 3 illustrates the assessment of cognitive ability in the paradigm of a recognition object (OR), demonstrating that (2S, 3R) -N- (2 - ((3-pyridinyl) methyl) -1-azabicyclo [2.2.2] oct-3 -yl) benzofuran-2-carboxamide has a positive effect at a dose of 0.3 and 1 mg / kg, introduced ip (intraperitoneally), p * <0.5.

Figure 4 illustrates the cognitive assessment in the OR paradigm, demonstrating that (2S, 3R) -N- (2 - ((3-pyridinyl) methyl) -1-azabicyclo [2.2.2] oct-3-yl) benzofuran -2-carboxamide, administered orally, has a positive effect in a wide range of concentrations (0.3-10 mg / kg), p * <0.5.

Figure 5 illustrates the effect of (2S, 3R) -N- (2 - ((3-pyridinyl) methyl) -1-azabicyclo [2.2.2] oct-3-yl) benzofuran-2-carboxamide, administered intraperitoneally, prevention of cognitive deficiency caused by MK-801, also known as disocylpin, a commercially available non-competitive NMDA receptor antagonist, in the OR task.

Figure 6 illustrates that the average time spent on object A relative to object B in the OR task in the test for the vehicle-treated group 30 minutes, 6 hours or 24 hours after the final subacute administration (oral) did not differ significantly (p = 0.17, p = 0.35 and p = 0.12, respectively). Alternatively, 30 minutes, 2 hours, 6 hours or 18 hours after the final subacute administration of (2S, 3R) -N- (2 - ((3-pyridinyl) methyl) -1-azabicyclo [2.2.2] oct-3- il) benzofuran-2-carboxamide at a dose of 0.3 mg / kg, subjects spent significantly more time (P <0.05), examining object B (new) than object A (familiar). Moreover, at time 2 hours (75%) and 6 hours (71%), the recognition index was significantly better in animals treated with (2S, 3R) -N- (2 - ((3-pyridinyl) methyl) -1- azabicyclo [2.2.2] oct-3-yl) benzofuran-2-carboxamide at a dose of 0.3 mg / kg, compared with the recognition index (54%) in the vehicle-treated group 30 minutes after the final administration.

Figure 7 illustrates the assessment of cognitive ability in the paradigm of a labyrinth with an angular lever (RAM). (2S, 3R) -N- (2 - ((3-pyridinyl) methyl) -1-azabicyclo [2.2.2] oct-3-yl) benzofuran-2-carboxamide (0.1, 0.3 and 1, 0 mg / kg) was administered orally 30 minutes before daytime classes. An improvement in the solution of the problem was evident in the group receiving 0.3 mg / kg (2S, 3R) -N- (2 - ((3-pyridinyl) methyl) -1-azabicyclo [2.2.2] oct-3-yl) benzofuran -2-carboxamide during the second week of administration.

FIG. 8 illustrates a study of antipsychotic activity, measured as hyperactive behavior caused by dopamine overstimulation, showing that (2S, 3R) -N- (2 - ((3-pyridinyl) methyl) -1-azabicyclo [2.2.2] oct -3-yl) benzofuran-2-carboxamide (0.3 and 1.0 mg / kg; subcutaneously) reduces locomotor hyperactivity caused by apomorphine (1.0 mg / kg) after subcutaneous administration to rats.

Figure 9 illustrates an antipsychotic assessment, prepulse inhibition, indicating that apomorphine deficiency is reversible in pretreatment of (2S, 3R) -N- (2 - ((3-pyridinyl) methyl) -1-azabicyclo [2.2.2] oct-3-yl) benzofuran-2-carboxamide after subcutaneous administration.

10A illustrates the results of an X-ray diffraction crystallographic analysis of (2S, 3R) -N- (2 - ((3-pyridinyl) methyl) -1-azabicyclo [2.2.2] oct-3-yl) benzofuran-2-carboxamide, established absolute stereochemistry of a substance. The compound shown is a partially hydrated hydrochloride salt, as shown by a completely disordered chloride anion and partial placement of a water molecule in an asymmetric cell.

10B illustrates the results of an X-ray diffraction crystallographic analysis of (2S, 3R) -N- (2 - ((3-pyridinyl) methyl) -1-azabicyclo [2.2.2] oct-3-yl) benzofuran-2-carboxamide monohydrochloride, which established the absolute stereochemistry of the substance depicted with the atom numbering scheme for reference. A downward view of the crystallographic b-axis of the unit cell is presented. Intermolecular hydrogen bonds are shown by dashed lines.

On figa illustrates the results of x-ray crystallographic analysis of p-chlorobenzoate (2R, 3R) -N- (2 - ((3-pyridinyl) methyl) -1-azabicyclo [2.2.2] oct-3-yl) benzofuran-2- carboxamide, which established the absolute stereochemistry of a substance.

11B illustrates the results of an X-ray diffraction crystallographic analysis of p-chlorobenzoate (2R, 3R) -N- (2 - ((3-pyridinyl) methyl) -1-azabicyclo [2.2.2] oct-3-yl) benzofuran-2- carboxamide, which established the absolute stereochemistry of a substance, depicted with an atom numbering scheme for reference.

12 illustrates a complete chromatogram characterizing the four stereoisomers of N- (2 - ((3-pyridinyl) methyl) -1-azabicyclo [2.2.2] oct-3-yl) benzofuran-2-carboxamide, where 2S, 3R- the isomer shows a peak with a retention time of 5.3 minutes, 2R, 3S-isomer shows a peak with a retention time of 7.3 minutes, 2R, 3R-isomer shows a peak with a retention time of 8.2 minutes and the 2S, 3S-isomer shows a peak with time retention of 12.4 minutes. As described in this invention, the mobile phase required in the analysis to ensure adequate separation leads to a hexane: ethanol: di-n-butylamine composition in a ratio of 60: 40: 0.2, at a flow rate of 1.0 ml / min, at column temperature 20 ° C and UV detection at 270 nm.

Figure 13 shows a powder X-ray diffraction pattern of (2S, 3R) -N- (2 - ((3-pyridinyl) methyl) -1-azabicyclo [2.2.2] oct-3-yl) benzofuran-2-carboxamide monohydrochloride, illustrating how observed (lighter) and calculated peaks (darker). In both cases, the nature of the distribution of the peaks is consistent with respect to the values of 2θ, and the minimal difference in the intensity and width of the peaks may be due to the resolution of the devices and preferred orientation effects. As described in this invention, an additional minimal difference may be due to the temperature shift, since the observed data were obtained at room temperature, and the calculation data were obtained for the structure at 120 K.

On Fig shows a powder x-ray of monotosylate (2S, 3R) -N- (2 - ((3-pyridinyl) methyl) -1-azabicyclo [2.2.2] oct-3-yl) benzofuran-2-carboxamide.

DETAILED DESCRIPTION OF THE INVENTION

Particular compound, (2S, 3R) -N- (2 - ((3-pyridinyl) methyl) -1-azabicyclo [2.2.2] oct-3-yl) benzofuran-2-carboxamide having affinity (Ki value <1 nM) and selectivity for the α7 NNR subtype, is effective in animal models of consciousness (increased consciousness) and psychosis (antipsychotic effect).

In one aspect, the present invention relates to (2S, 3R) -N- (2 - ((3-pyridinyl) methyl) -1-azabicyclo [2.2.2] oct-3-yl) benzofuran-2-carboxamide or a pharmaceutically acceptable thereof salt. Another aspect relates to (2S, 3R) -N- (2 - ((3-pyridinyl) methyl) -1-azabicyclo [2.2.2] oct-3-yl) benzofuran-2-carboxamide essentially in pure form or to its pharmaceutically acceptable salt. A further aspect relates to (2S, 3R) -N- (2 - ((3-pyridinyl) methyl) -1-azabicyclo [2.2.2] oct-3-yl) benzofuran-2-carboxamide substantially free of (2S , 3S), (2R, 3S) or (2R, 3R) isomers, or a pharmaceutically acceptable salt thereof.

Additionally, another aspect relates to stereochemically enriched (2S, 3R) -N- (2 - ((3-pyridinyl) methyl) -1-azabicyclo [2.2.2] oct-3-yl) benzofuran-2-carboxamide or its pharmaceutically acceptable salt. In one embodiment, the enantiomeric and / or diastereomeric excess is 90% or more. In one embodiment, the enantiomeric and / or diastereomeric excess is 95% or more. In one embodiment, the enantiomeric and / or diastereomeric excess is 98% or more. In one embodiment, the enantiomeric and / or diastereomeric excess is 99% or more. In one embodiment, the enantiomeric and / or diastereomeric excess is 99.5% or more.

Another aspect of the present invention relates to a salt of (2S, 3R) -N- (2 - ((3-pyridinyl) methyl) -1-azabicyclo [2.2.2] oct-3-yl) benzofuran-2-carboxamide with an acid, where the acid is selected from hydrochloric acid, sulfuric acid, phosphoric acid, maleic acid, p-toluenesulfonic acid, galactaric (mucous) acid, D-mandelic acid, D-tartaric acid, methanesulfonic acid, R- and S-10-camphorsulfonic acid, ketoglutaric acid acid or hippuric acid. In one embodiment, the stoichiometric ratio of (2S, 3R) -N- (2 - ((3-pyridinyl) methyl) -1-azabicyclo [2.2.2] oct-3-yl) benzofuran-2-carboxamide to acid is 2: 1, 1: 1 or 1: 2. In one embodiment, the stoichiometric ratio is 1: 1. One embodiment of the present invention is (2S, 3R) -N- (2 - ((3-pyridinyl) methyl) -1-azabicyclo [2.2.2] oct-3-yl) benzofuran-2-carboxamide hydrochloride or a hydrate thereof or a solvate, including partial hydrates or solvates. A further embodiment is (2S, 3R) -N- (2 - ((3-pyridinyl) methyl) -1-azabicyclo [2.2.2] oct-3-yl) benzofuran-2-carboxamide monohydrochloride or a hydrate or solvate thereof including partial hydrates or solvates.

Another aspect of the present invention relates to (2S, 3R) -N- (2 - ((3-pyridinyl) methyl) -3-amino-1-azabicyclo [2.2.2] octane.

Another aspect of the present invention relates to a pharmaceutical composition comprising a compound of the present invention and one or more pharmaceutically acceptable carriers.

Another aspect of the present invention relates to a method for treating or preventing a central nervous system disorder, inflammation, pain or neovascularization, comprising administering a compound of the present invention. In one embodiment, a violation of the central nervous system is characterized by a change in the normal release of the neurotransmitter. In one embodiment, a central nervous system disorder is selected from mild cognitive impairment associated with age-related memory impairment, preschool dementia, early onset of Alzheimer's disease, senile dementia, Alzheimer's type of dementia, Alzheimer's disease, dementia with Levi bodies, dementia due to microinfarction, with AIDS dementia, HIV dementia, multiple cerebral infarction, parkinsonism, Parkinson's disease, Peak's disease, progressive supranuclear palsy, chorus Huntington, tardive dyskinesia, hyperkinesia, mania, disorders associated with attention deficit hyperactivity disorder, attention deficit, anxiety, depression, dyslexia, schizophrenia, cognitive dysfunction in schizophrenia, depression, obsessive-compulsive disorders and Tourette's syndrome. In one embodiment, a central nervous system disorder is selected from Alzheimer's disease, mania, attention deficit disorder, attention deficit hyperactivity disorder, anxiety, dyslexia, schizophrenia, cognitive dysfunction in schizophrenia, depression, obsessive-compulsive disorder, or Tourette’s syndrome.

Another aspect of the present invention includes the use of a compound of the present invention for the manufacture of a medicament for treating or preventing a central nervous system disorder, inflammation, pain or neovascularization. In one embodiment, a violation of the central nervous system is characterized by a change in the normal release of the neurotransmitter. In one embodiment, a central nervous system disorder is selected from mild cognitive impairment associated with age-related memory impairment, preschool dementia, early onset of Alzheimer's disease, senile dementia, Alzheimer's type of dementia, Alzheimer's disease, dementia with Levi bodies, dementia due to microinfarction, with AIDS dementia, HIV dementia, multiple cerebral infarction, parkinsonism, Parkinson's disease, Peak's disease, progressive supranuclear palsy, chorus Huntington, tardive dyskinesia, hyperkinesia, mania, disorders associated with attention deficit hyperactivity disorder, attention deficit, anxiety, depression, dyslexia, schizophrenia, cognitive dysfunction in schizophrenia, depression, obsessive-compulsive disorders and Tourette's syndrome. In one embodiment, a central nervous system disorder is selected from Alzheimer's disease, mania, attention deficit disorder, attention deficit hyperactivity disorder, anxiety, dyslexia, schizophrenia, cognitive dysfunction in schizophrenia, depression, obsessive-compulsive disorder, or Tourette’s syndrome.

Another aspect of the present invention relates to a compound of the present invention for use in the treatment or prevention of central nervous system disorder, inflammation, pain or neovascularization. In one embodiment, a violation of the central nervous system is characterized by a change in the normal release of the neurotransmitter. In one embodiment, a central nervous system disorder is selected from mild cognitive impairment associated with age-related memory impairment, preschool dementia, early onset of Alzheimer's disease, senile dementia, Alzheimer's type of dementia, Alzheimer's disease, dementia with Levi bodies, dementia due to microinfarction, with AIDS dementia, HIV dementia, multiple cerebral infarction, parkinsonism, Parkinson's disease, Peak's disease, progressive supranuclear palsy, chorus Huntington, tardive dyskinesia, hyperkinesia, mania, disorders associated with attention deficit hyperactivity disorder, attention deficit, anxiety, depression, dyslexia, schizophrenia, cognitive dysfunction in schizophrenia, depression, obsessive-compulsive disorders and Tourette's syndrome. In one embodiment, a central nervous system disorder is selected from Alzheimer's disease, mania, attention deficit disorder, attention deficit hyperactivity disorder, anxiety, dyslexia, schizophrenia, cognitive dysfunction in schizophrenia, depression, obsessive-compulsive disorder, or Tourette’s syndrome.

In the above methods and applications in one embodiment of the invention, effective doses are between 1 mg and 10 mg over a 24 hour period.

Another aspect of the present invention relates to a method for producing (2S, 3R) -N- (2 - ((3-pyridinyl) methyl) -1-azabicyclo [2.2.2] oct-3-yl) benzofuran-2-carboxamide or its pharmaceutically acceptable salt by sequential dynamic optical separation and stereoselective reductive amination of (2 - ((3-pyridinyl) methyl) -1-azabicyclo [2.2.2] oct-3-one.

The scope of the present invention includes all combinations of aspects, embodiments, and advantages described herein.

Commercial development of potential drugs involves many stages, including scaling of chemical synthesis and purification, identification of optimal salt forms and the like. In the formulation of drug compositions, the drug substance is preferably in a form that is convenient to work with and handle. The analysis includes commercial viability as well as stability upon receipt. In addition, when receiving medicinal compositions, it is important that a reliable, reproducible and constant profile of the concentration of the drug after its administration to the patient is provided in the blood plasma.

Chemical stability, solid stability, and shelf life of active ingredients are also very important factors. The drug substance and the composition containing it should preferably be able to be stored efficiently for a substantial period of time without significantly changing the physicochemical characteristics of the active component (for example, its chemical composition, density, hygroscopicity and solubility). In addition, it is also important to be able to obtain the drug in a form that is as pure as possible. These features of the invention are discussed in more detail below.

I. Compounds

The compound of the present invention is (2S, 3R) -N- (2 - ((3-pyridinyl) methyl) -1-azabicyclo [2.2.2] oct-3-yl) benzofuran-2-carboxamide, shown below as a compound A, or pharmaceutically acceptable salt forms of compound A.

Compound A

The racemic compound - N- (2 - ((3-pyridinyl) methyl) -1-azabicyclo [2.2.2] oct-3-yl) benzofuran-2-carboxamide, its synthesis and use in medical applications are described in US Pat. No. 6953855 , Mazurova and others included in this description by reference.

The racemic N- (2 - ((3-pyridinyl) methyl) -1-azabicyclo [2.2.2] oct-3-yl) benzofuran-2-carboxamide is a high affinity ligand for the α7 subtype of the neuronal nicotinic receptor (NNR). Racemic N- (2 - ((3-pyridinyl) methyl) -1-azabicyclo [2.2.2] oct-3-yl) benzofuran-2-carboxamide contains two asymmetrically substituted carbon atoms. Thus, the racemic N- (2 - ((3-pyridinyl) methyl) -1-azabicyclo [2.2.2] oct-3-yl) benzofuran-2-carboxamide exists in the form of four stereoisomeric forms, namely, (S, S), (S, R), (R, R) and (R, S). The isomer (S, R), namely (2S, 3R), is a compound A.

It was previously assumed that the dominant stereoisomeric forms obtained in the reported syntheses, including US Patent No. 6953855, are characterized by a relative cis configuration at the 2 and 3 positions of the 1-azabicyclo [2.2.2] octane (quinuclidine) ring. In other words, it was believed that the cis diastereomer ((2R, 3R) and (2S, 3S), a pair of enantiomers) are predominant forms that are formed upon receipt of the reported methods. This definition of cis synthesis predominance was based on (I) comparing the ¹ H constants ^{of the} spin-spin interaction of hydrogen nuclei at the 2 and 3 positions of the quinuclidine ring and the isolated diastereomeric (cis and trans) intermediates with the spin-spin coupling constants reported in the literature ; and in (II) the expected stereochemical yield of synthetic chemistry used to obtain a mixture of compounds by analogy with literature, with reference to Warawa et al., J. Med. Chem. 18 (6): 587-593 (1975); and Viti et al., Tetrahedron Lett., 35 (32): 5939-5942 (1994), both of which are incorporated herein by reference. Thus, it was expected that a cis configuration would form. As such, biological testing using the racemate led to results that were attributed as inherent in the dominant cis configuration.

It has now been established, using X-ray diffraction analysis of crystalline salt forms and analogs, that the dominant diastereomer formed in the initial synthesis was, in fact, a trans-diastereomer. In addition, it was found that two enantiomers with relative trans stereochemistry, namely (2S, 3R) and (2R, 3S), differ significantly in their ability to interact with the α7 NNR subtype. Compound (2S, 3R) Compound A has greater activity.

Using additional analysis, it was found that compound A has pharmacological properties that differ from (i) the properties of each of the other three stereoisomers taken separately; (ii) the properties of the mixture of all four stereoisomers, namely, the racemate; and (iii) properties of other ligands of the NNR α7 receptor reported in the literature.

(2S, 3R) -N- (2 - ((3-pyridinyl) methyl) -1-azabicyclo [2.2.2] oct-3-yl) benzofuran-2-carboxamide (compound A) is a highly selective complete NNR receptor agonist α7 with a remarkably low EC ₅₀ (for activation) and a good difference between the EC ₅₀ and IC ₅₀ (for residual inhibition), providing functional agonism in a wide range of therapeutically useful concentrations.

II. Scalable Synthesis of (2S, 3R) -N- (2 - ((3-pyridinyl) methyl) -1-azabicyclo [2.2.2] oct-3-yl) benzofuran-2-carboxamide

Specific synthetic stages vary in their ability to scale. It has been found that reactions are difficult to scale for a variety of reasons, including safety considerations, the high cost of reagents, difficult processing or purification, energy parameters of the reaction (thermodynamics or kinetics), and reaction yield.

The synthesis of (2S, 3R) -N- (2 - ((3-pyridinyl) methyl) -1-azabicyclo [2.2.2] oct-3-yl) benzofuran-2-carboxamide described in this invention was used to obtain kilogram amounts substances, and component reactions were carried out on a multi-kilogram scale in high yield.

The scalable synthesis used both dynamic optical separation of the racemizable ketone (2 - ((3-pyridinyl) methyl) -1-azabicyclo [2.2.2] octan-3-one) and stereoselective reduction of the imine derivative (R) -α-methylbenzylamine (reductive amination) of isomeric ketone. The synthetic sequence described herein is easily scalable and does not include chromatographic purification steps.

III. Obtaining new salt forms of (2S, 3R) -N- (2 - ((3-pyridinyl) methyl) -1-azabicyclo [2.2.2] oct-3-yl) benzofuran-2-carboxamide

(2S, 3R) -N- (2 - ((3-pyridinyl) methyl) -1-azabicyclo [2.2.2] oct-3-yl) benzofuran-2-carboxamide in the form of a free base is an amorphous powder with very limited solubility in water. The free base will react with both inorganic and organic acids to produce certain acid addition salts that have physical and chemical properties that are beneficial for pharmaceutical compositions, including, but not limited to, crystallinity, solubility in water, and stability . The stoichiometry of the salts of the present invention may vary.

Depending on the method by which the salts described in this invention are formed, they may have a crystalline structure in which the solvents that are present during the preparation of the salt are occluded. Thus, salts can exist as hydrates and other solvates with variable solvent stoichiometry relative to (2S, 3R) -N- (2 - ((3-pyridinyl) methyl) -1-azabicyclo [2.2.2] oct-3-yl) benzofuran-2-carboxamide.

The method of obtaining salt forms may vary. The preparation of the salt forms of (2S, 3R) -N- (2 - ((3-pyridinyl) methyl) -1-azabicyclo [2.2.2] oct-3-yl) benzofuran-2-carboxamide usually includes

(i) mixing a free base or a solution of a free base, namely (2S, 3R) -N- (2 - ((3-pyridinyl) methyl) -1-azabicyclo [2.2.2] oct-3-yl) benzofuran-2 β-carboxamide, in a suitable solvent with an acid without solvent, or as a solution of an acid in a suitable solvent;

(iia) cooling the resulting salt solution, if necessary, in order to cause precipitation; or

(iib) adding a suitable “anti” solvent to cause precipitation; or

(iic) evaporating the first solvent and adding a new solvent and repeating either step (iia) or step (iib); and

(iii) filtering and collecting the resulting salt.

In one embodiment, (2S, 3R) -N- (2 - ((3-pyridinyl) methyl) -1-azabicyclo [2.2.2] oct-3-yl) benzofuran-2-carboxamide is stereochemically enriched. In one embodiment, the enantiomeric and / or diastereomeric excess is 90% or more. In one embodiment, the enantiomeric and / or diastereomeric excess is 95% or more. In one embodiment, the enantiomeric and / or diastereomeric excess is 98% or more. In one embodiment, the enantiomeric and / or diastereomeric excess is 99% or more. In one embodiment, the enantiomeric and / or diastereomeric excess is 99.5% or more.

The stoichiometry, solvent mixture, solute concentration, and temperature used may vary. Illustrative solvents that can be used to prepare or recrystallize salt forms include, but are not limited to, ethanol, methanol, isopropyl alcohol, acetone, ethyl acetate, and acetonitrile.

Examples of suitable pharmaceutically acceptable salts include inorganic acid addition salts such as chloride, bromide, sulfate, phosphate and nitrate; organic acid addition salts such as acetate, galactarate, propionate, succinate, lactate, glycolate, malate, tartrate, citrate, maleate, fumarate, methanesulfonate, p-toluenesulfonate and ascorbate; and salts with amino acids such as aspartate and glutamate. Salts in some cases may be hydrates or solvates with ethanol. Illustrative salts are provided as described in US Pat. Nos. 5,597,919, Dull et al., 5616716, Dull et al., And 5663356, Ruecroft et al., Each of which is incorporated herein by reference.

Screening of the free base salts of (2S, 3R) -N- (2 - ((3-pyridinyl) methyl) -1-azabicyclo [2.2.2] oct-3-yl) benzofuran-2-carboxamide showed that although they can be obtained many salts with pharmaceutically acceptable acids, only some of these salts have acceptable properties for commercial production. The ability to predict characteristics illustrating a commercially viable salt, respectively, does not exist. Acids that produce salts that are crystalline, namely salts that exhibit some degree of crystallinity depending on the process by which they are obtained, include hydrochloric acid, sulfuric acid, phosphoric acid, p-toluenesulfonic acid, galactaric (mucus) acid , D-mandelic acid, D-tartaric acid, methanesulfonic acid, R- and S-10-camphorsulfonic acid, maleic acid, ketoglutaric acid and hippuric acid. Among these salts, each of the salts of hydrochloric acid, phosphoric acid, maleic acid, and p-toluenesulfonic acid exhibited additional desirable properties, including high melting points, good solubility in water, and low hygroscopicity. Such characteristics for these salts were unexpected.

IV. Pharmaceutical Compositions

The pharmaceutical compositions of the present invention include the salts described herein, in a pure state or as a composition in which the compounds are combined with any other pharmaceutically compatible product that may be inert or physiologically active. The resulting pharmaceutical compositions can be used to prevent a condition or disorder in a subject subject to such a condition or disorder, and / or to treat a subject suffering from a given condition or disorder. The pharmaceutical compositions described herein include a compound of the present and / or pharmaceutically acceptable salts thereof.

The manner in which the compounds are administered may vary. The compositions are preferably administered orally (for example, in liquid form in a solvent, such as an aqueous or non-aqueous liquid, or in a solid carrier). Preferred compositions for oral administration include pills, tablets, capsules, caplets, syrups and solutions, including hard gelatine capsules and time-release capsules. Standard excipients include binders, fillers, colorants, solubilizers and the like. Formulation of the compositions may be in the form of a single preparative dosage form or in the form of multiple doses or subunits. Preferred compositions are in liquid or semi-solid form. Compositions comprising a liquid pharmaceutically inert carrier, such as water or other pharmaceutically compatible liquids, or semi-solids, can be used. The use of such liquids and semi-solids is well known to those skilled in the art.

The compositions can also be administered by injection, i.e. intravenously, intramuscularly, subcutaneously, intraperitoneally, intraarterially, intrathecal and intracerebroventricular. Intravenous administration is the preferred method of injection. Suitable injection vehicles are well known to those skilled in the art and include 5% dextrose solutions, saline and phosphate buffered saline. The drug product can also be administered as an infusion or injection (for example, as a suspension or as an emulsion in a pharmaceutically acceptable liquid or mixture of liquids).

Drugs can also be administered using other agents, for example, by rectal administration. Preparations used for rectal administration, such as suppositories, are well known to those skilled in the art. A drug product can also be administered by inhalation (for example, as an aerosol, either nasally or using delivery devices of the type described in US Pat. No. 4,922,901 to Brooks et al., The disclosure of which is incorporated herein in its entirety); externally (for example, in the form of a lotion); transdermally (for example, using a transdermal patch) or iontophoretic; or by sublingual or buccal administration. Although it is possible to administer the compound as a bulk of the active chemical, it is preferred that the drug product is in the form of a pharmaceutical composition or preparation for an effective and efficient administration.

Illustrative methods for administering the compounds will be apparent to those skilled in the art. The applicability of these formulations may depend on the particular composition used and the particular subject receiving the treatment. These preparations may contain a liquid carrier, which may be oily, aqueous, emulsified, or contain certain solvents suitable for a given route of administration.

The compositions can be administered by fractional administration or at a gradual, continuous, constant or controlled speed to a warm-blooded animal (e.g., a mammal, such as a mouse, rat, cat, rabbit, dog, pig, cow or monkey), but they are mainly administered to humans. In addition, the time during the day and the number of times per day when the pharmaceutical composition is administered may vary.

Other suitable methods for administering the compounds of the present invention are described in US Pat. No. 5,604,231 to Smith et al., The contents of which are incorporated herein by reference.

In an embodiment of the present invention and as will be appreciated by those skilled in the art, the compound of the present invention can be administered in combination with other drug compounds. For example, the compound of this invention can be used in combination with other NNR ligands (such as varenicline), antioxidants (such as free radical scavengers), antimicrobials (such as penicillin antibiotics), and antiviral agents (such as zidovudine nucleoside analogues and acyclovir), anticoagulants (such as warfarin), anti-inflammatory drugs (such as NSAIDs), antipyretic drugs, analgesics, anesthetics (such as those used by the surgeon ii), acetylcholinesterase inhibitors (such as donepezil and galantamine), antipsychotic drugs (such as haloperidol, clozapine, olanzapine and quetipine), immunosuppressants (such as cyclosporine and methotrexate), neuroprotective agents such as steroids (steroids, steroids (steroids, steroids) like dexamethasone, predisone and hydrocortisone), vitamins, minerals, dietary supplements, antidepressants (such as imipramine, fluoxetine, paroxetine, escitalopram, sertraline, venlafaxine and duloxetine), anxiolytic antidepressants (such as alprazolam and baspirone), anticonvulsants (such as phenytoin and gabapentin), vasodilators (such as prazosin and sildenafil), mood stabilizers (such as valproate and aripiprazole), anti-cancer drugs (such as antiproliferative drugs) drugs (such as atenolol, clonidine, amlopidine, verapamil and olmesartan), laxatives, stool thinners, diuretics (such as furosemide), antispasmodics (such like dicyclomine), antidiscinetic drugs, and antiulcer drugs (such as esomeprazole).

The compounds of the present invention can be used alone or in combination with other therapeutic agents. Such a combination of pharmaceutically active agents can be administered together or separately and, when administered separately, the administration can be carried out simultaneously or sequentially in any order. The amounts of compounds or agents and the relative distribution of administration over time will be selected to achieve the desired therapeutic effect. Administration in combination can be carried out as concomitant administration in the form of (1) a single pharmaceutical composition comprising many compounds; or (2) individual pharmaceutical compositions, each of which includes one of the compounds. Alternatively, the combination can be administered individually in a sequential manner, where one drug is administered first and the other second, or vice versa. Such sequential administration can be close in time or separated in time. The compounds of the present invention can be used to treat many disorders and conditions and, as such, the compounds of the present invention can be used in combination with many other suitable therapeutic agents used to treat or prevent these disorders or conditions.

A suitable dose of the compound is an amount effective to prevent the onset of symptoms of the disorder or to treat certain symptoms of the disorder that the patient suffers from. As noted, the terms "effective amount", "therapeutic amount" or "effective dose" means that the amount is sufficient to provide the desired pharmacological or therapeutic effect, thus leading to an effective prevention or treatment of the disorder.

In the treatment of central nervous system disorder, an effective amount of a compound is an amount sufficient to pass through the subject’s blood-brain barrier to bind to relevant receptor sites in the subject’s brain and to modulate the activity of the relevant NNR subtypes (for example, by secreting a neurotransmitter, thereby effectively preventing or treating the disorder ) An example of preventing a disorder is manifested in a temporary delay in the onset of the onset of symptoms of the disease. An example of the treatment of a disorder is manifested in reducing the symptoms associated with the disorder, or alleviating the relapse of the symptoms of the disorder. Preferably, an effective amount is sufficient to obtain the desired result, but not sufficient to cause tangible side effects.

The effective dose may vary depending on factors such as the condition of the patient, the severity of the symptoms of the disease, and the manner in which the pharmaceutical composition is administered. For human patients, an effective dose of typical compounds usually requires the administration of a compound in an amount sufficient to modulate the activity of the relevant NNR, but the amount should be insufficient to cause any significant effect in the skeletal muscle or ganglion. The effective dose, of course, will vary from patient to patient, but, in general, includes amounts starting from the moment the action on the central nervous system or other desired therapeutic actions appears, but lower than the amount at which muscle effects are observed.

The compounds described herein, when used in effective amounts in accordance with the methods described herein, can provide a degree of prevention of progression or amelioration of symptoms or treatment, and to some extent, relapse of CNS disorders or other disorders. Effective amounts of these compounds are usually below the concentration threshold required to cause any noticeable side effects, such as those related to skeletal muscle or the ganglion. Compounds can be administered in a therapeutic window in which some CNS disorders and other disorders are treated, and certain similar actions are excluded. Ideally, an effective dose of the compounds described herein is sufficient to provide the desired effect on the disorder, but insufficient (i.e., not at such a high enough level) to cause unwanted side effects. Preferably, the compounds are administered in a dosage that is effective for treating CNS disorders or other disorders, but less than, often less than 1/5, and often less than 1/10, the amount necessary to cause certain similar actions in what any significant degree.

Most preferably, effective dosages are at very low concentrations at which maximum action occurs with minimal side effects. Typically, an effective dose of such compounds usually requires administration of the compound in an amount of less than 5 mg / kg of patient weight. Often, the compounds of the present invention are administered in an amount of less than about 1 mg / kg of patient weight and usually less than about 100 μg / kg of patient weight, but more often in the range of about 10 μg to less than 100 μg / kg of patient weight. The above effective doses usually represent the amount administered as a single dose or as one or more doses administered over a 24-hour period.

For human patients, an effective dose of conventional compounds usually requires administration of the compound in an amount of at least about 1, often at least about 10, and more often at least about 100 mg / 24 hours / patient. For human patients, an effective dose of conventional compounds usually requires the administration of a compound in an amount that usually does not exceed about 500, often does not exceed about 400, and more often does not exceed about 300 mg / 24 hours / patient. Additionally, the compositions are preferably administered in such an effective dose that the concentration of the compound in the patient’s blood plasma usually does not exceed 50 ng / ml, often does not exceed 30 ng / ml, and more often does not exceed 10 ng / ml. In one embodiment of the present invention, an effective dose is between about 1 and 10 mg over a 24-hour period.

IV. Method for the use of pharmaceutical compositions

As used herein, the term “agonist” refers to a substance that stimulates its binding partner, usually a receptor. Stimulation is defined in the context of a particular analysis, or it may be apparent from the literature discussed here, which compares a factor or substance that is accepted as an “agonist” or “antagonist” of a particular binding partner under essentially the same circumstances as recognized by those skilled in the art. . Stimulation can be determined by increasing a specific effect or function that is caused by the interaction of an agonist or partial agonist with a binding partner and may include allosteric effects.

As used herein, the term “antagonist” refers to a substance that inhibits its binding partner, usually a receptor. Inhibition is defined in the context of a specific assay, or it can be apparent from the literature discussed here, which compares a factor or substance that is accepted as an “agonist” or “antagonist” of a particular binding partner under essentially the same circumstances as recognized by those skilled in the art . Inhibition can be determined by reducing a specific effect or function that is caused by the interaction of an antagonist with a binding partner and may include allosteric effects.

As used herein, the term "partial agonist" or "partial antagonist" refers to a substance that provides a level of stimulation or inhibition, respectively, of its binding partner, which is not completely or completely agonistic or antagonistic, respectively. It will be understood that stimulation and, correspondingly, inhibition are defined essentially for any substance or category of substances that will be defined as agonists, antagonists or partial agonists.

As used herein, the term “internal activity” or “effectiveness” refers to a certain measure of the biological effectiveness of the partner binding complex. With respect to receptor pharmacology, the context in which intrinsic activity or potency will be determined will depend on the context of the partner binding complex (e.g., receptor / ligand) and consideration of activity related to a particular biological result. For example, under certain circumstances, internal activity may vary depending on the secondary messenger system involved. See Hoyer D and Boddeke H., Trends Pharmacol. Sci. 14 (7): 270-5 (1993), incorporated herein by reference for such indications. Where such contextually specific evaluations are significant, and to what extent they may be significant in the context of the present invention will be apparent to one of ordinary skill in the art.

As used herein, receptor modulation includes agonism, partial agonism, antagonism, partial antagonism, or inverse agonism of the receptor.

As used herein, neurotransmitters whose release is mediated by the compounds described herein include, but are not limited to, acetylcholine, dopamine, norepinephrine, serotonin, and glutamate, and the compounds described herein function as modulators of the CNS subtype α7 NNR.

As used herein, the term “prevention” or “prophylaxis” includes any degree of reducing the progression or delaying the onset of a disease, disorder or condition. The term includes providing a protective effect in relation to a particular disease, disorder or condition, as well as alleviating the recurrence of the disease, disorder or condition. Thus, in another aspect, the invention relates to a method for treating a subject at risk of developing or recurring a disorder mediated by NNR or nAChR. The compounds and pharmaceutical compositions of the invention can be used to achieve a beneficial therapeutic or prophylactic effect, for example, in a subject with central nervous system dysfunction.

As noted above, the compounds of the present invention in the form of a free base and salt modulate the subtype of the α7 NNR characteristic of the central nervous system and can be used to prevent or treat various conditions or disorders, including conditions or disorders of the central nervous system, in subjects who have or are at risk the occurrence of such conditions or disorders by modulating the α7 NNR. These compounds have the ability to selectively bind to α7 NNR and exhibit nicotine pharmacology, for example, act as agonists, partial agonists, antagonists, as described. For example, the compounds of the present invention, when administered in effective amounts to patients in need thereof, provide some degree of prevention of the development of a central nervous system disorder, namely, provide a protective effect, alleviate the symptoms of a central nervous system disorder or ameliorate the relapse of a central nervous system disorder, or a combination of these.

The compounds of the present invention can be used to treat or prevent those types of conditions and disorders for which other types of nicotine compounds have been proposed or have been shown to be useful as drugs. See, for example, the references listed earlier in this specification, as well as Williams et al., Drug News Perspec. 7 (4): 205 (1994), Arneric et al., CNS Drug Rev. 1 (1): 1-26 (1995), Arneric et al., Exp. Opin. Invest. Drugs 5 (1): 79-100 (1996); Bencherif et al., J. Pharmacol. Exp. Ther. 279: 1413 (1996), Lippiello et al., J. Pharmacol. Exp. Ther. 279: 1422 (1996), Damaj et al., J. Pharmacol. Exp. Ther. 291: 390 (1999); Chiari et al., Anesthesiology 91: 1447 (1999), Lavand'homme and Eisenbach, Anesthesiology 91: 1455 (1999), Holladay et al., J. Med. Chem. 40 (28): 4169-94 (1997), Bannon et al., Science 279: 77 (1998), PCT WO 94/08992, PCT WO 96/31475, PCT WO 96/40682 and US Pat. Nos. 5,583,140, Bencherif et al., 5597919, Dull et al., 5604231, Smith et al., and 5852041, Cosford et al., the description of which regarding such therapeutic indications is incorporated herein by reference.

The compound and their pharmaceutical compositions can be used to treat or prevent a variety of central nervous system disorders, including neurodegenerative disorders, neuropsychiatric disorders, neurological disorders and addictions. The compounds and their pharmaceutical compositions can be used to treat or prevent cognitive deficits and dysfunctions associated with age and other factors; impaired attention and dementia, including those that occur under the influence of infectious agents or metabolic disorders; to provide neuroprotection; for the treatment of convulsions and multiple cerebral infarction; for the treatment of mood disorders, manic and bad habits-related behavioral reactions; to provide pain relief; for controlling inflammation, such as mediated by cytokines and nuclear factor kappa B; for the treatment of inflammatory disorders; to provide pain relief; for the treatment of metabolic disorders, such as diabetes or metabolic syndrome; and for the treatment of infections as anti-infective agents for the treatment of bacterial, fungal and viral infections.

CNS disorders

Disorders, diseases, and conditions for the treatment or prophylaxis of which the compounds and pharmaceutical compositions of the present invention can be used include age-related memory impairment (AAMI), mild cognitive impairment (MCI), age-related cognitive decline (ARCD), preschool dementia, early onset of Alzheimer's disease, senile dementia, Alzheimer's type dementia, Alzheimer's disease, cognitive impairment without dementia (CIND), dementia with Lewy bodies (DTL), HIV dementia, complex connection AIDS-related dementia, multi-infarct dementia, Down syndrome, head trauma, traumatic brain injury (TBI), dementia due to trauma, Creutzfeldt-Jakob disease and prion diseases, stroke, ischemia, attention deficit disorder, attention deficit hyperactivity disorder, dyslexia, schizophrenia, disorder of the schizophrenic form, schizophrenia-like disorder of the affective type, cognitive dysfunction in schizophrenia, cognitive deficiency in schizophrenia, such as memory deficiency, including working memory, organization functions, extra possessions, vigilance, information processing and learning related to schizophrenia dementia (any of mild, moderate or severe), parkinsonism, including Parkinson's disease, Parkinson's dementia parkinsonism, Gaum’s dementia, frontotemporal dementia Parkinson’s type (FTDP), H Peak (Niemann-Pick), Huntington’s disease, Huntington’s chorea, tardive dyskinesia, hyperkinesia, progressive supranuclear palsy, progressive supranuclear paresis, tired leg syndrome, multiple sclerosis, amyotrophic lateral sclerosis (ALS), motor neuron disease (MND), multiple systemic atrophy (MSA), corticobasal degeneration, Guillaume-Barré syndrome (GBS), chronic inflammatory demyelinating polyneuropathy (CIDP), epilepsy, autosomal dominant fronto-temporal temporalis , anxiety, depression, premenstrual dysphoria, panic disorders, bulimia, anorexia, narcolepsy, excessive drowsiness in the daytime, bipolar disorders, generalized anxiety, obsessive-compulsive disorder Ie, outbursts of rage, violation of opposition disobedience, Tourette’s syndrome, autism, drug addiction and chronic alcoholism, smoking addiction, obesity, cachexia, psoriasis, lupus, acute angiocholitis, aphthous stomatitis, ulcers, asthma, ulcerative colitis, inflammatory bowel disease, K disease , postoperative intestinal obstruction, spasmodic dystonia, diarrhea, constipation, reservoir ileitis, pancreatitis, viral pneumonia, arthritis, including rheumatoid arthritis and osteoarthritis, endotoxemia, sepsis, atherosclerosis, idiopathic fib Lake lung, acute pain, chronic pain, neuropathy, incontinence, diabetes and neoplasia.

Cognitive impairment or dysfunction may be associated with psychiatric disorders or conditions, such as schizophrenia and other psychotic disorders, including, but not limited to, psychotic disorder, schizophrenic disorder, schizoaffective disorder, delusional disorder, short-term psychotic disorder, acute psychotic disorder, and psychotic disorder due to one or more general medical conditions, various types of dementia and other cognitive impairment, including but not limited to With this, mild cognitive impairment, pre-dementia, Alzheimer's disease, senile dementia, Alzheimer's dementia, age-related memory impairment, dementia with Levi bodies (DTL), vascular dementia, AIDS-related dementia complex, dyslexia, parkinson's disease Parkinson's, cognitive impairment and dementia in Parkinson's disease, cognitive impairment in multiple sclerosis, cognitive impairment caused by traumatic brain damage, dementia due to other common m medical conditions, anxiety conditions, including, but not limited to, panic state without agoraphobia, panic state with concomitant agoraphobia, agoraphobia without a history of panic disorder, specific phobias, social phobia, obsessive-compulsive syndrome, post-traumatic stress disorder, acute stress disorder, generalized stress condition and generalized anxiety due to general medical condition, mood disorders, including, but not limited to, profound depression, mental depression, bipolar depression, bipolar mania, type I bipolar disorder, depression related to mania, depressive or mixed episodes, type II bipolar disorder, cyclothymic disorder and mood disorders due to general medical condition, sleep disturbances, including but not limited to , insomnia, primary insomnia, primary increased drowsiness, narcolepsy, parasomnia, disturbances associated with nightmares, disturbances in the horror of sleep and disturbances associated with walking during sleep, mental extinction, learning disabilities, impaired motor skills, communication disorders, impairment of growth, attention deficit and impairment of destructive behavior, impairment of attention deficit hyperactivity disorder associated with impaired attention, feeding and nutritional disorders infants, children or adults, tics, elimination disorders, substance-related disorders, including but not limited to drug abuse, drug intoxication, discontinuation drug addiction alcohol-related disorders amphetamine-related or amphetamine-like compounds caffeine-related disorders marijuana-related disorders cocaine-related disorders inhalant-related disorders nicotine-related disorders nicotine-related disorders opioid-related disorders disorders associated with phencyclidine or phencyclidine-like disorders; and disorders associated with sedative, hypnotic or anxiolytic agents, n Rushen person, including but not limited to, obsessive-compulsive personality disorder and impulse-control disorders.

Symptoms of schizophrenia are usually divided into three categories: positive, negative and cognitive. Positive symptoms may also be referred to as “psychotic” symptoms and include mania and hallucinations. Positive symptoms are referred to as apparent symptoms. Negative symptoms include emotional depression or lack of expression, inability to start and continue active activity, speech that is brief and devoid of content, lack of pleasure and interest in active activity. Negative symptoms relate to a lack of certain characteristics that would otherwise be present in a healthy person. Cognitive symptoms relate to thinking processes. Cognitive symptoms include a lack of cognitive abilities such as memory, including working memory, organizing function, attention, vigilance, information processing and learning, with reference to Sharma et al., Cognitive Function in Scizophrenia: Deficits, Functional Consequences and Future Treatment, Psychiatr . Clin. N. Am. 26 (2003) 25-40, incorporated herein by reference. Schizophrenia also affects mood. Although many individuals affected by schizophrenia are prone to depression, some also have marked mood swings and even bipolar-like conditions.

The above conditions and abnormalities are discussed in more detail, for example, in the American Psychiatric Association: Diagnostic and Statistical Manual of Mental Disorders, fourth edition, revised, Washington, DC, American Psychiatric Association, 2000; included in this description by reference to identify such conditions and disorders. This guide can also be mentioned for a more detailed description of symptoms and diagnostic features associated with the use, abuse and dependence on various substances.

Preferably, the treatment or prevention of diseases, disorders and conditions occurs without noticeable side effects, including, for example, a significant increase in blood pressure and heart rate, a significant negative effect on the gastrointestinal tract and a significant effect on the muscles of the skeleton.

It is believed that the compounds of the present invention, when used in effective amounts, modulate the activity of α7 NNR without noticeable interaction with other subtypes of nicotinic receptors that are characteristic of the human ganglion, as demonstrated by a lack of ability to induce nicotine function in adrenaline chromaffin tissue or skeleton muscle, further demonstrated by a lack of ability induce nicotine function in preparations of cells expressing nicotinic receptors of muscle ty a. Thus, it can be assumed that these compounds are able to treat or prevent diseases, disorders and conditions without causing noticeable side effects associated with the activity of ganglion and neuromuscular sites. Thus, it is believed that the administration of the compounds will provide a therapeutic window providing treatment for certain diseases, disorders and conditions in which some side effects are excluded. That is, it is believed that an effective dose of the compound is sufficient to provide the desired effect on the disease, disorder or condition, but it is believed to be insufficient, namely not at such a high level to cause unwanted side effects.

Thus, the present invention relates to the use of a compound of the present invention or a pharmaceutically acceptable salt thereof for use in therapy, such as the therapy described above.

In another aspect, the present invention relates to the use of a compound of the present invention or a pharmaceutically acceptable salt thereof for the manufacture of a medicament for use in the treatment of a central nervous system disorder, such as the disorder, disease or condition described above.

Inflammation

It is known that the nervous system, mainly through the vagus nerve, regulates the inherent immune response by inhibiting the release of macrophage tumor necrosis factor (TNF). This physiological mechanism is known as the “cholinergic anti-inflammatory pathway” (see, for example, Tracey, “The inflammatory reflex”, Nature 420: 853-9 (2002), incorporated herein by reference). Excessive inflammation and the synthesis of tumor necrosis factor cause morbidity and even mortality in many diseases. These diseases include, but are not limited to, endotoxemia, rheumatoid arthritis, osteoarthritis, psoriasis, asthma, atherosclerosis, idiopathic pulmonary fibrosis, and inflammatory bowel disease.

Inflammatory conditions that can be treated or prevented by the administration of the compounds described in this invention include, but are not limited to, chronic and acute inflammation, psoriasis, endotoxemia, gout, acute pseudogout, acute gouty arthritis, arthritis, rheumatoid arthritis, osteoarthritis , polymyositis, dermatomyositis, ankylosing spondylitis, Stille disease, the onset of adult Stille disease, allograft rejection, chronic transplant rejection, asthma, atherosclerosis h, mononuclear-phagocytic dependent lung damage, idiopathic pulmonary fibrosis, atopic dermatitis, chronic obstructive pulmonary disease, adult respiratory distress syndrome, acute pulmonary syndrome with sickle cell disease, inflammatory bowel disease, Crohn's disease, ulcerative colitis, acute angiocholitis, aphthosis (thrush), reservoir ileitis, glomerulonephritis, lupus nephritis, thrombosis, and graft-versus-host reaction.

Bacterial inflammatory responses

and / or viral infection

Many bacterial and / or viral infections (e.g., meningitis, hepatitis, and nephritis) are associated with side effects from the formation of toxins and the body's natural response to bacteria or viruses and / or toxins. As discussed above, the body's response to infections often involves the formation of a significant amount of TNF and / or other cytokines. Overexpression of such cytokines can cause significant harm, such as septic shock (when the bacterium is sepsis), endotoxic shock, urosepsis and toxic shock syndrome.

Cytokine expression is mediated by NNR and can be inhibited by the administration of agonists or partial agonists of these receptors. Those compounds described herein are agonists or partial agonists of these receptors, and therefore, can be used to minimize the inflammatory response associated with bacterial infection, as well as viral and fungal infections. Examples of such bacterial infections include carbuncles, botulism, and sepsis. Some of these compounds may also have antibacterial properties.

These compounds can also be used as adjuvant therapy in combination with existing methods of treating bacterial, viral and fungal infections, such as antibiotics, antiviral drugs and antifungal drugs. Antitoxins can also be used to bind toxins produced by infectious agents, and to pass through the body as bound toxins without causing an inflammatory response. Examples of antitoxins are described, for example, in US patent No. 6310043, Bundle et al., Incorporated herein by reference. Other agents effective against bacteria and other toxins may be effective, and their therapeutic effect may be supplemented by co-administration with the compounds described herein.

Pain

The compounds can be used to treat and / or prevent pain, including acute, neurological, inflammatory, neuropathic and chronic pain. The analgesic activity of the compounds described herein can be demonstrated on a model of persistent inflammatory pain and neuropathic pain, as described in published US patent application No. 20010056084 A1 (Allgeier et al.), Incorporated herein by reference, which demonstrates rat hyperalgesia inflammatory pain models with Freund's complete adjuvant; and mechanical hyperalgesia of neuropathic pain in a mouse model of partial ligation of the sciatic nerve.

An analgesic effect is suitable for the treatment of pain of various origins or etiologies, in particular for the treatment of inflammatory pain and related hyperalgesia, neuropathic pain and related hyperalgesia, chronic pain (e.g. severe chronic pain, postoperative pain and pain associated with various conditions including cancer, angina pectoris, renal or biliary colic, menstruation, migraine and gout). Inflammatory pain can be of various origins, including arthritis and rheumatoid disease, synovitis of tendon sheaths and vasculitis. Neuropathic pain includes trigeminal or herpetic neuralgia, diabetic neuropathic pain, burning pain, lower back pain and deafferentation syndromes such as rupture of the brachial plexus.

Neovascularization

The α7 NNR receptor is associated with neovascularization. Inhibition of neovascularization, for example, by administration of antagonists (or at certain dosages of partial agonists), α7 NNR can treat or prevent conditions characterized by unwanted neovascularization or angiogenesis. Such conditions may include those characterized by inflammatory angiogenesis and / or ischemia-induced angiogenesis. Neovascularization associated with tumor growth can also be inhibited by administering compounds such as those described in this invention that function as antagonists or partial α7 NNR agonists.

Specific antagonism of the specific activity of α7 NNR reduces the angiogenic response to inflammation, ischemia and neoplasia. Guidance on suitable animal model systems for evaluating the compounds described herein can be found, for example, in Heeschen C. et al., “A novel angiogenic pathway mediated by non-neuronal nicotinic acetylcholine receptors”, J. Clin. Invest. 110 (4): 527-36 (2002), incorporated herein by reference to describe α7-specific inhibition of angiogenesis and cellular (in vitro) and animal models of modeling angiogenic activity significant for human disease, especially a model of a Levi tumor of the lung human (in vivo, in mice see, in particular, pages 529 and 532-533).

Illustrative types of tumors that can be treated using the compounds described herein include NSCLC (non-small cell lung carcinoma), ovarian cancer, pancreatic cancer, breast carcinoma, colon carcinoma, rectal carcinoma, lung carcinoma, oropharyngeal carcinoma, carcinoma of the throat , carcinoma of the esophagus, carcinoma of the stomach, carcinoma of the pancreas, carcinoma of the liver, carcinoma of the gallbladder, carcinoma of the bile duct, carcinoma of the small intestine, carcinoma of the urinary tract, renal cell carcinoma, bladder carcinoma, urothelial carcinoma, female genital tract carcinoma, cervical carcinoma, uterine carcinoma, ovarian carcinoma, trophoblastic tumor, gestational trophoblastic disease (related to pregnancy), male genital tract carcinoma, prostate carcinoma, seminal carcinoma testicular carcinoma, embryo tumor, endocrine carcinoma, thyroid carcinoma, adrenal carcinoma, pituitary carcinoma, skin carcinoma, hemanglioma, melanoma, sarco we, bone and soft tissue sarcoma, Kaposi’s sarcoma, brain tumors, nerve tumors, eye tumors, brain lining tumors, astrocytomas, gliomas, glioblastomas, retinoblastomas, neuromas, neuroblastomas, schwannomas, meningiomas, dense tumors arising from hematopoietic malignancies ( such as leukemia, chlorleukemia, plasmacytomas and mushroom-like plaques and tumors of the mucosa and lymphoma / leukemia of skin T cells), and dense tumors arising from lymphoma.

The compounds can also be administered in combination with other forms of anticancer therapy, including co-administration with antineoplastic antitumor agents such as cisplatin, adriamycin, daunomycin and the like and / or anti-VEGF (vascular endothelial growth factor) agents, such as are known in this area.

Compounds can be administered in such a way that they will target the tumor site. For example, the compounds can be administered in microspheres, microparticles, or liposomes conjugated to various antibodies that direct particles to the tumor. In addition, the compounds may be present in microspheres, microparticles, or liposomes that are sized to fit through arteries and veins, but which get stuck in the capillary bed surrounding the tumor, and inject the compounds into the tumor locally. Such drug delivery devices are known in the art.

Other violations

In addition to treating CNS disorders, inflammation, neovascularization and pain, the compounds of the present invention can also be used to prevent or treat certain other conditions, diseases and disorders, in which NNRs play a role. Examples include autoimmune disorders such as lupus, disorders associated with the release of cytokines, secondary cachexia after infection (for example, as observed in AIDS, AIDS-related complex and neoplasia), metabolic disorders, including type I diabetes, type II diabetes, metabolic syndrome , obesity or hypercalcemia, pemphigus, urinary incontinence, retinal diseases, infectious diseases, myasthenia gravis, Eaton-Lambert syndrome, high blood pressure, osteoporosis, narrowing of blood vessels, dilatation of blood vessels, heart arrhythmia, bulimia, anorexia, as well as the indications indicated in published PCT application WO 98/25619, which is incorporated into this description by reference on the subject of such violations. The compounds of this invention can also be used to treat convulsions, such as those symptomatic of epilepsy, and to treat conditions such as syphilis and Creutzfeldt-Jakob disease.

Diagnostic use

The compounds can be used in diagnostic compositions, such as probes, in particular if they are modified in such a way as to include suitable labels. Probes can be used, for example, to determine the relative number and / or function of specific receptors, especially a subtype of α7 receptors. For this purpose, a label of a radioactive isotope, such as ¹¹ C, ¹⁸ F, ⁷⁶ Br, ¹²³ I or ¹²⁵ I, is preferably introduced into the compounds of the present invention.

Introduced compounds can be detected using known detection methods suitable for the label used. Examples of detection methods include positron emission tomography (PET) and single photon emission computed tomography (SPECT). The radioactive labels described above can be used with PET (e.g. ¹¹ C, ¹⁸ F or ⁷⁶ Br) and SPECT (e.g. ¹²³ I) imaging with half-lives of approximately 20.4 minutes for ¹¹ C, approximately 109 minutes for ¹⁸ F , approximately 13 hours for ¹²³ I, and approximately 16 hours for ⁷⁶ Br. To visualize selected receptor subtypes at concentrations not reaching saturation, high specific activity is desirable. Doses administered are usually below the toxic range and provide high contrast images. It is expected that the compounds can be used at non-toxic levels. Dose determination is carried out by a method known to the person skilled in the art of imaging using radioactive labels. See, for the introduction of such compounds, for example, US patent No. 5969144, London et al., Incorporated into this invention by reference.

Compounds can be administered using known methods. See, for example, US patent No. 5969144, London et al., As noted, included in this invention by reference to questions regarding such an introduction. The compounds can be administered in prescription compositions that include other ingredients, such as, for example, those types of ingredients that are used in formulating diagnostic compositions. The compounds used in accordance with the implementation of the present invention, most preferably used in forms with a high degree of purity. See, for such an analysis, US Pat. No. 5,853,696 to Elmalch et al., Incorporated herein by reference.

After the administration of the compounds to a subject (eg, a human), the presence of such a compound in the subject's body can be visually displayed and quantified using suitable methods to detect the presence, amount and functionality of the selected NNR subtypes. In addition to humans, the compounds can also be administered to animals such as mice, rats, dogs, and monkeys. SPECT and PET imaging can be performed using any suitable method and instrument. For a description of illustrative visualization techniques, see Villemagne et al., In Arneric et al. (Eds.) Neuronal Nicotinic Receptors: Pharmacology and Therapeutic Opportunities, 235-250 (1998) and US Patent No. 5853696, Elmalch et al., Each of which is incorporated herein by reference.

Radiolabeled compounds bind with high affinity to selective NNR subtypes (e.g., α7) and preferably exhibit negligible non-specific binding to other subtypes of nicotinic cholinergic receptors (e.g., those subtypes of receptors that are associated with the musculature and nerve nodes (ganglia)). As such, the compounds can be used as agents for non-invasive imaging of subtypes of nicotinic cholinergic receptors in the body of a subject, in particular in the brain, for diagnosis associated with a variety of diseases and disorders of the central nervous system.

In one aspect, diagnostic compositions can be used in a method for diagnosing a disease in a subject, such as a human. The method comprises administering to such a patient a detectable label compound as described herein and detecting binding of such a compound to selected NNR subtypes (e.g., α7 receptor subtype). Specialists in the use of diagnostic tools such as PET and SPECT can use the radiolabeled compounds described herein to diagnose a large number of conditions and disorders, including conditions and disorders associated with dysfunction of the central and autonomic nervous system. Such disorders include many diseases and disorders of the central nervous system, including Alzheimer's disease, Parkinson's disease and schizophrenia. You can evaluate these and other illustrative diseases and disorders, including those indicated in US patent No. 5952339, Bencherif et al., Incorporated into this description by reference.

In another aspect, the diagnostic compositions can be used in a method for controlling selective nicotinic receptor subtypes in a subject, such as a human patient. The method comprises administering to such a patient a detectable label compound as described herein, and detecting the binding of such a compound to selected nicotinic receptor subtypes, namely the α7 receptor subtype.

Receptor binding

The compounds of this invention can be used as comparison ligands in binding assays for compounds that bind to NNR subtypes, in particular the α7 receptor subtype. For this purpose, a radioactive label is preferably introduced into the compounds of the invention using a radioactive isotope fragment, such as ³ N or ¹⁴ C.

V. Synthetic examples

The following synthetic examples are provided to illustrate the present invention and should not be construed as limiting the scope of the invention. In these experiments, all parts and percentages are by weight unless otherwise indicated. All solutions are aqueous solutions, unless otherwise indicated.

Example 1

Small-scale synthesis of (2S, 3R) -N- (2 - ((3-pyridinyl) methyl) -1-azabicyclo [2.2.2] octan-3-yl) benzofuran-2-carboxamide (compound A) and its enantiomer (2R, 3S) -N- (2 - ((3-pyridinyl) methyl) -1-azabicyclo [2.2.2] octan-3-yl) benzofuran-2-carboxamide

2 - ((3-Pyridinyl) methylene) -1-azabicyclo [2.2.2] octan-3-one

Potassium hydroxide (56 g, 0.54 mol) was dissolved in methanol (420 ml). 3-Quinuclidinone hydrochloride (75 g, 0.49 mol) was added and the mixture was stirred for 30 minutes at ambient temperature. 3-Pyridinecarboxaldehyde (58 g, 0.54 mol) was added and the mixture was stirred for 16 hours at ambient temperature. During this time, the reaction mixture turned yellow, and a solid precipitate formed on the walls of the flask. Solids were scraped off the walls and large pieces were destroyed. With rapid stirring, water (390 ml) was added. When the solids were dissolved, the mixture was cooled to 4 ° C and kept overnight. The crystals were collected by filtration, washed with water and dried in air, obtaining 80 g of a yellow solid. A second portion (8 g) was obtained by concentrating the filtrate to ~ 10% of its original volume and keeping it under cooling to 4 ° C overnight. Both portions were clean enough for subsequent conversion (88 g, 82% yield).

2 - ((3-Pyridinyl) methyl) -1-azabicyclo [2.2.2] octan-3-one

2 - ((3-Pyridinyl) methylene) -1-azabicyclo [2.2.2] octan-3-one (20 g, 93 mmol) was suspended in methanol (200 ml) and treated with 46 ml of 6M hydrochloric acid. 10% palladium-on-charcoal (1.6 g) was added and the mixture was shaken at a hydrogen pressure of 25 psi for 16 hours. The mixture was filtered through diatomaceous earth and the solvent was removed from the filtrate by evaporation on a rotary evaporator. This gave crude 2 - ((3-pyridinyl) methyl) -1-azabicyclo [2.2.2] octan-3-one hydrochloride as a white gum (20 g), which was subsequently treated with 2M sodium hydroxide (50 ml) and chloroform ( 50 ml) and stirred for one hour. The chloroform layer was separated and the aqueous phase was treated with 2M sodium hydroxide (~ 5 ml, enough to raise the pH to 10) and a saturated aqueous solution of sodium chloride (25 ml). The aqueous mixture was extracted with chloroform (3 × 10 ml) and the combined chloroform extract was dried (anhydrous magnesium sulfate) and concentrated by evaporation on a rotary evaporator. The residue (18 g) was dissolved in warm ether (320 ml) and cooled to 4 ° C. The white solid was filtered off, washed with a small portion of cold ether and dried in air. Concentration of the filtrate to ~ 10% of its initial volume and cooling to 4 ° C gave a second portion. A combined yield of 16 g (79%) was obtained.

3-amino-2 - ((3-pyridinyl) methyl) -1-azabicyclo [2.2.2] octane

To a stirred solution of 2 - ((3-pyridinyl) methyl) -1-azabicyclo [2.2.2] octan-3-one (3.00 g, 13.9 mmol) in dry methanol (20 ml) in a nitrogen atmosphere was added 1M a solution of zinc chloride in ether (2.78 ml, 2.78 mmol). After stirring at ambient temperature for 30 minutes, this mixture was treated with solid ammonium formate (10.4 g, 167 mmol). After stirring for another hour at ambient temperature, solid sodium cyanoborohydride (1.75 g, 27.8 mmol) was added portionwise. The reaction mixture was then stirred at ambient temperature overnight and the reaction was stopped by adding water (~ 5 ml). After quenching the reaction mixture was distributed between 5M sodium hydroxide (10 ml) and chloroform (20 ml). The aqueous layer was extracted with chloroform (20 ml) and the combined organic layers were dried (sodium sulfate), filtered and concentrated. This resulted in a residue of 2.97 g of yellow gum. GLC mass spectrometric (GCMS) analysis showed that the product was a 1: 9 mixture of cis and trans amines along with traces of the corresponding alcohol (98% recovery by total weight).

(2R, 3S) and (2S, 3R) -3-amino-2 - ((3-pyridinyl) methyl) -1-azabicyclo [2.2.2] octane

Di-p-toluoyl-D-tartaric acid (5.33 g, 13.8 mmol) was added to a stirred solution of the crude 3-amino-2 - ((3-pyridinyl) methyl) -1-azabicyclo [2.2.2] octane (6.00 g, 27.6 mmol cis / trans in a 1: 9 ratio) in methanol (20 ml). After completion of dissolution, the clear solution was then concentrated to obtain a solid mass by evaporation on a rotary evaporator. The solid was then dissolved in a minimum amount of boiling methanol (~ 5 ml). The solution was slowly cooled, initially to ambient temperature (1 h), then kept for ~ 4 hours at 5 ° C and, finally, at -5 ° C overnight. The precipitated salt was collected by filtration with suction of the solvent in vacuo and recrystallized from 5 ml of methanol. Air drying gave 1.4 g of a white solid, which was partitioned between chloroform (5 ml) and 2M sodium hydroxide (5 ml). The chloroform layer and 5 ml of the chloroform extract of the aqueous layer were combined, dried (anhydrous sodium sulfate) and concentrated to give a colorless oil (0.434 g). Enantiomeric purity for a given free base was determined by converting part of it to the corresponding N- (tert-butoxycarbonyl) -L-prolinamide, for which diastereomeric purity (98%) was then analyzed using LCMS.

The mother liquor after initial crystallization was made basic (~ pH 11) with 2 M sodium hydroxide and extracted twice with chloroform (10 ml). The chloroform extracts were dried (anhydrous sodium sulfate) and concentrated to give an oil. This amine (3.00 g, 13.8 mmol) was dissolved in methanol (10 ml) and treated with di-p-toluoyl-L-tartaric acid (2.76 g, 6.90 mmol). The mixture was heated in order to facilitate dissolution and then slowly cooled to -5 ° C, leaving at this temperature overnight. The precipitate was collected by filtration with suction of the solvent in vacuo, recrystallized from methanol and dried. This gave a residue of 1.05 g of a white solid. This salt was converted to the free base (yield = 0.364 g) and enantiomeric purity (97%) was evaluated using the proline method, as described above for the other enantiomer.

Trans Enantiomer A N- (2 - ((3-pyridinyl) methyl) -1-azabicyclo [2.2.2] octan-3-yl) benzofuran-2-carboxamide

Diphenylchlorophosphate (0.35 ml, 0.46 g, 1.7 mmol) was added dropwise to a solution of benzofuran-2-carboxylic acid (0.28 g, 1.7 mmol) and triethylamine (0.24 ml, 0.17 g, 1.7 mmol) in dry dichloromethane (5 ml). After stirring at ambient temperature for 30 minutes, a solution of (2S, 3R) -3-amino-2 - ((3-pyridinyl) methyl) -1-azabicyclo [2.2.2] octane (0.337 g, 1.55 mmol) was added ) (which was obtained from the salt of di-p-toluolyl-D-tartaric acid) and triethylamine (0.24 ml, 0.17 g, 1.7 mmol) in dry dichloromethane (5 ml). The reaction mixture was stirred overnight at ambient temperature and then treated with 10% sodium hydroxide solution (1 ml). The biphasic mixture was separated and the organic layer was concentrated on a Genevac centrifugal evaporator. The residue was dissolved in methanol (6 ml) and purified by HPLC on a C18 silica gel column using an acetonitrile / water gradient containing 0.05% trifluoroacetic acid as an eluent. Concentration of the selected fractions, the distribution of the obtained residue between chloroform and saturated aqueous sodium bicarbonate and evaporation of chloroform gave 0.310 g (42% yield) of white powder (95% purity according to GLC mass spectrometry). ¹ H NMR (300 MHz, CDCl ₃ ) δ 8.51 (d, 1H), 8.34 (dd, 1H), 7.66 (d, 1H), 7.58 (dt, 1H), 7.49 (d, 1H), 7.44 (s, 1H), 7.40 (dd, 1H), 7.29 (t, 1H), 7.13 (dd, 1H), 6.63 (d, 1H) 3.95 (t, 1H), 3.08 (m, 1H), 2.95 (m, 4H), 2.78 (m, 2H), 2.03 (m, 1H), 1.72 ( m, 3H); 1.52 (m, 1H).

It was later determined using chiral chromatographic analysis that this substance (trans-enantiomer A) is identical to a substance whose absolute configuration is 2S, 3R (determined by X-ray diffraction analysis).

Trans Enantiomer B N- (2 - ((3-pyridinyl) methyl) -1-azabicyclo [2.2.2] octan-3-yl) benzofuran-2-carboxamide

Diphenylchlorophosphate (96 μl, 124 mg, 0.46 mmol) was added dropwise to a solution of benzofuran-2-carboxylic acid (75 mg, 0.46 mmol) and triethylamine (64 μl, 46 mg, 0.46 mmol) in dry dichloromethane (1 ml). After stirring at ambient temperature for 45 minutes, a solution of (2R, 3S) -3-amino-2 - ((3-pyridinyl) methyl) -1-azabicyclo [2.2.2] octane (0.10 g, 0, 46 mmol) (which was obtained from the salt of di-p-toluolyl-L-tartaric acid) and triethylamine (64 μl, 46 mg, 0.46 mmol) in dry dichloromethane (1 ml). The reaction mixture was stirred overnight at ambient temperature and then treated with 10% sodium hydroxide solution (1 ml). The biphasic mixture was separated and the organic layer and the chloroform extract (2 ml) of the aqueous layer were concentrated using a rotary evaporator. The residue was dissolved in methanol and purified by HPLC on a C18 silica gel column using an acetonitrile / water gradient containing 0.05% trifluoroacetic acid as an eluent. Concentration of the selected fractions, distribution of the obtained residue between chloroform and saturated aqueous sodium bicarbonate and evaporation of chloroform gave 82.5 mg (50% yield) of white powder. The NMR spectrum was identical to the spectrum obtained for the 2S, 3R isomer. Since the direct precursor of this substance (trans-enantiomer B) is enantiomeric to the direct precursor of the 2S, 3R compound (trans-enantiomer A), the absolute configuration of trans-enantiomer B was assumed to be 2R, 3S.

Example 2. Large-scale synthesis of (2S, 3R) -N- (2 - ((3-pyridinyl) methyl) -1-azabicyclo [2.2.2] octan-3-yl) benzofuran-2-carboxamide and p-toluenesulfonate salt ( 2S, 3R) -N- (2 - ((3-pyridinyl) methyl) -1-azabicyclo [2.2.2] oct-3-yl) -1-benzofuran-2-carboxamide.

2 - ((3-Pyridinyl) methylene) -1-azabicyclo [2.2.2] octan-3-one

3-Quinuclidinone hydrochloride (8.25 kg, 51.0 mol) and methanol (49.5 L) were added, under a nitrogen atmosphere, to a 100 L glass reaction flask equipped with a mechanical stirrer, temperature sensor and reflux condenser. Potassium hydroxide (5.55 kg, 99.0 mol) was added through a funnel for the powder over about 30 minutes, which led to an increase in the temperature of the reaction mixture from 50 ° C to 56 ° C. 3-pyridinecarboxaldehyde (4.80 kg, 44.9 mol) was added to the reaction mixture over about 2 hours. The resulting mixture was stirred at 20 ° C ± 5 ° C for at least 12 hours as monitoring the progress of the reaction using thin layer chromatography (TLC). After completion of the reaction, the reaction mixture was filtered through a sintered glass filter, and the filter cake was washed with methanol (74.2 L). The filtrate was concentrated, transferred to a reaction flask, and water (66.0 L) was added. The suspension was stirred for at least 30 minutes, filtered and the filter cake was washed with water (90 L) until the pH of the solution after washing reached 7-9. The solid was dried in vacuo at 50 ° C ± 5 ° C for at least 12 hours to give 8.58 kg (89.3%) of 2 - ((3-pyridinyl) methylene) -1-azabicyclo [2.2.2] octan-3-one.

(2S) -2-p-Toluoyl-D-tartrate salt - ((3-pyridinyl) methyl) -1-azabicyclo [2.2.2] octan-3-one

2 - ((3-Pyridinyl) methylene) -1-azabicyclo [2.2.2] octan-3-one (5.40 kg, 25.2 mol) and methanol (40.5 L) in a 72 L s reaction flask inert atmosphere, equipped with a mechanical stirrer, a temperature sensor, a regulating low-pressure gas system and a pressure measuring device. The space above the reaction mixture was filled with nitrogen and the mixture was stirred until a clear yellow solution was obtained. 10% palladium-on-charcoal (50% wet) (270 g) was added to the flask. The atmosphere from the reactor was removed using a vacuum pump and the space above the reaction mixture was replaced with hydrogen to a pressure of 10-20 inches of water. The removal of the gas atmosphere from the flask and the supply of hydrogen under pressure were repeated 2 more times, leaving the reactor under pressure of hydrogen gas in 20 inches of water after the third supply of gas under pressure. The reaction mixture was stirred at 20 ° C ± 5 ° C for at least 12 hours and the progress of the reaction was monitored by TLC. After completion of the reaction, the suspension was filtered through a layer of Celite®545 (1.9 kg) on a porous glass filter and the filter cake was washed with methanol (10.1 L). The filtrate was concentrated to obtain a semi-solid, which was transferred, under nitrogen atmosphere, into a 100 L reaction flask equipped with a mechanical stirrer, reflux condenser and temperature sensor. The semi-solid was dissolved in ethanol (57.2 L) and di-p-toluoyl-D-tartaric acid (DTTA) (9.74 kg, 25.2 mol) was added. The stirred reaction mixture was heated at the boil under reflux for at least 1 hour and stirred additionally for at least 12 hours as the reaction mixture was cooled to a temperature between 15 ° C and 30 ° C. The suspension was filtered through a bench top filter and the filter cake was washed with ethanol (11.4 L). The product was dried in vacuo at ambient temperature to give 11.6 kg (72% yield, 59.5% pure factorizable) of the di-p-toluoyl-D-tartrate salt (2S) -2 - ((3-pyridinyl) methyl ) -1-azabicyclo [2.2.2] octan-3-one.

(2S, 3R) -3-amino-2 - ((3-pyridinyl) methyl) -1-azabicyclo [2.2.2] octane di-p-Toluoyl-D-tartrate salt

Water (46.25 l) and sodium bicarbonate (4.35 kg, 51.8 mol) were added to a 200 l flask. After complete dissolution, dichloromethane (69.4 L) was added. (2S) -2 - ((3-pyridinyl) methyl) -1-azabicyclo [2.2.2] octan-3-one (11.56 kg, 19.19 mol) di-p-toluoyl-D-tartrate salt was added. and the reaction mixture was stirred for 2 minutes to 10 minutes. The layers were allowed to separate for at least 2 minutes (additional water (20 L) was added if necessary for layer distribution). The organic phase was removed and dried over anhydrous sodium sulfate. Dichloromethane (34.7 L) was added to the remaining aqueous phase, and the suspension was stirred for 2 minutes to 10 minutes. The layers were allowed to separate for 2 minutes to 10 minutes. Again, the organic phase was removed and dried over anhydrous sodium sulfate. The extraction of the aqueous phase with dichloromethane (34.7 L) was repeated one more time as described above. Samples obtained in each extraction were analyzed using chiral HPLC analysis. Sodium sulfate was removed by filtration, and the filtrates were concentrated to obtain (2S) -2 - ((3-pyridinyl) methyl) -1-azabicyclo [2.2.2] octan-3-one (4.0 kg) as a solid.

(2S) -2 - ((3-Pyridinyl) methyl) -1-azabicyclo [2.2.2] octan-3-one (3.8 kg) was transferred under nitrogen atmosphere into a 100 L clean glass reaction flask equipped with a mechanical stirrer and temperature sensor. Anhydrous tetrahydrofuran (7.24 L) and (+) - (R) -α-methylbenzylamine (2.55 L, 20.1 mol) were added. Titanium (IV) isopropoxide (6.47 L, 21.8 mol) was added to the stirred reaction mixture over 1 hour. The reaction mixture was stirred under nitrogen for at least 12 hours. Ethanol (36.17 L) was added to the reaction mixture. The reaction mixture was cooled to a temperature below -5 ° C and sodium borohydride (1.53 kg, 40.5 mol) was added portionwise, keeping the temperature of the reaction mixture below 15 ° C (this addition took several hours). The reaction mixture was then stirred at 15 ° C ± 10 ° C for a minimum of 1 hour. The progress of the reaction was monitored by HPLC and after completion of the reaction (as indicated by the presence of less than 0.5% of the remaining (2S) -2 - ((3-pyridinyl) methyl) -1-azabicyclo [2.2.2] octane-3- she) 2M sodium hydroxide (15.99 L) was added and the mixture was stirred for at least 10 minutes. The reaction mixture was filtered through a pad of Celite®545 on a tabletop funnel. The filter cake was washed with ethanol (15.23 L) and the filtrate was concentrated to give an oil.

The concentrate was transferred to a clean glass reaction flask with a capacity of 100 l, equipped with a mechanical stirrer and temperature sensor, in an inert atmosphere. Water (1 L) was added and the mixture was cooled to 0 ° C ± 5 ° C. To the mixture was added 2M hydrochloric acid (24 L) to adjust the pH of the mixture to pH 1. Then the mixture was stirred for at least 10 minutes and the aqueous phase was extracted with dichloromethane (3 × 15.23 L). The organic phases were dried over anhydrous sodium sulfate (2.0 kg), filtered and concentrated to give (2S, 3R) -N - ((1R) -phenylethyl) -3-amino-2- ((3-pyridinyl) methyl) - 1-azabicyclo [2.2.2] octane (4.80 kg, 84.7% yield).

(2S, 3R) -N - ((1R) -Phenylethyl) -3-amino-2 - ((3-pyridinyl) methyl) -1-azabicyclo [2.2.2] octane was transferred into a 22 L glass flask equipped with a mechanical stirrer and temperature sensor in an inert atmosphere. Water (4.8 L) was added and the stirred mixture was cooled to 5 ° C ± 5 ° C. Concentrated hydrochloric acid (2.97 L) was slowly added to the reaction flask, keeping the temperature of the mixture below 25 ° C. The resulting solution was transferred to a 72 L reaction flask containing ethanol (18 L), equipped with a mechanical stirrer, temperature sensor and reflux condenser, in an inert atmosphere. 10% palladium-on-charcoal (50% wet) (311.1 g) and cyclohexene (14.36 L) were added to the flask. The reaction mixture was heated at a temperature close to the reflux temperature for at least 12 hours and the progress of the reaction was monitored by TLC. After completion of the reaction, the reaction mixture was cooled to a temperature below 45 ° C and filtered through a pad of Celite® 545 (1.2 kg) on a funnel with a glass porous filter. The filter cake was washed with ethanol (3 L) and the filtrate was concentrated to give an aqueous phase. Water (500 ml) was added to the concentrated filtrate, and the combined aqueous layer was washed with methyl tert-butyl ether (MTBE) (2 × 4.79 L). 2M sodium hydroxide (19.5 L) was added to the aqueous phase to adjust the pH of the mixture to pH 14. The mixture was then stirred for at least 10 minutes. The aqueous phase was extracted with chloroform (4 × 11.96 L) and the combined organic phases were dried over anhydrous sodium sulfate (2.34 kg). The filtrate was filtered and concentrated to give (2S, 3R) -3-amino-2 - ((3-pyridinyl) methyl) -1-azabicyclo [2.2.2] octane (3.49 kg> than quantitative yield) as an oil.

(2S, 3R) -3-amino-2 - ((3-pyridinyl) methyl) -1-azabicyclo [2.2.2] octane was transferred into a clean 100-liter reaction flask equipped with a mechanical stirrer, reflux condenser and temperature sensor, in inert atmosphere. Ethanol (38.4 L) and di-p-toluoyl-D-tartaric acid (3.58 kg, 9.27 mol) were added. The reaction mixture was heated at mild reflux for at least 1 hour. The reaction mixture was then stirred for at least 12 hours as it cooled to a temperature between 15 ° C and 30 ° C. The resulting suspension was filtered and the filter cake washed with ethanol (5.76 L). The filter cake was transferred to a clean glass reaction flask with a capacity of 100 l, equipped with a mechanical stirrer, reflux condenser and temperature sensor, in an inert atmosphere. An ethanol / water solution was added in a ratio of 9: 1 (30.7 L) and the resulting suspension was heated at mild reflux for at least 1 hour. The reaction mixture was then stirred for at least 12 hours as it cooled to a temperature between 15 ° C and 30 ° C. The mixture was filtered and the filter cake washed with ethanol (5.76 L). The product was collected and dried in vacuo at 50 ° C ± 5 ° C for at least 12 hours to obtain 5.63 kg (58.1% yield) of dip-toluoyl-D-tartrate salt (2S, 3R) -3 amino-2 - ((3-pyridinyl) methyl) -1-azabicyclo [2.2.2] octane.

(2S, 3R) -N- (2 - ((3-pyridinyl) methyl) -1-azabicyclo [2.2.2] octan-3-yl) benzofuran-2-carboxamide

(2S, 3R) -3-amino-2 - ((3-pyridinyl) methyl) -1-azabicyclo [2.2.2] octane di-p-Toluoyl-D-tartrate salt (3.64 kg, 5.96 mol ) and a 10% aqueous solution of sodium chloride (14.4 L, 46.4 mol) were added to a 72 L glass reaction flask equipped with a mechanical stirrer in an inert atmosphere. 5M sodium hydroxide (5.09 L) was added to the stirred mixture to adjust the pH of the mixture to pH 14. The mixture was then stirred for at least 10 minutes. The aqueous solution was extracted with chloroform (4 × 12.0 L) and the combined organic layers were dried over anhydrous sodium sulfate (1.72 kg). The combined organic layers were filtered and the filtrate was concentrated to give (2S, 3R) -3-amino-2 - ((3-pyridinyl) methyl) -1-azabicyclo [2.2.2] octane (1.27 kg) as an oil.

(2S, 3R) -3-amino-2 - ((3-pyridinyl) methyl) -1-azabicyclo [2.2.2] octane was transferred into a 50 L glass reaction flask equipped with a mechanical stirrer in an inert atmosphere. Dichloromethane (16.5 L), triethylamine (847 ml, 6.08 mol), benzofuran-2-carboxylic acid (948 g, 5.85 mol) and O- hexafluorophosphate O- (benzotriazol-1-yl) - were added to the reaction mixture. N, N, N, 1-tetramethyluronium (HBTU) (2.17 kg, 5.85 mol). The mixture was stirred for at least 4 hours at ambient temperature and the progress of the reaction was monitored by HPLC. After completion of the reaction, a 10% aqueous potassium carbonate solution (12.7 L, 17.1 mol) was added to the reaction mixture, and the mixture was stirred for at least 5 minutes. The layers were separated and the organic phase was washed with 10% sodium chloride solution (12.7 L). The layers were separated and the organic phase was cooled to 15 ° C ± 10 ° C. 3M hydrochloric acid (8.0 L) was slowly added to the reaction mixture to adjust the pH of the mixture to pH 1. The mixture was then stirred for at least 5 minutes and the layers were allowed to distribute for at least 5 minutes. Solids were filtered using a bench top filter. The layers of the filtrate were separated and the aqueous phase and solids from the funnel were transferred to a reaction flask. 3M sodium hydroxide (9.0 L) was slowly added portionwise to the flask to adjust the pH of the mixture to pH 14. The aqueous phase was extracted with dichloromethane (2 × 16.5 L). The combined organic phases were dried over anhydrous sodium sulfate (1.71 kg). The mixture was filtered and the filtrate concentrated to give (2S, 3R) -N- (2 - ((3-pyridinyl) methyl) -1-azabicyclo [2.2.2] octan-3-yl) benzofuran-2-carboxamide (1.63 kg, 77.0% yield) as a yellow solid.

(2S, 3R) -N- (2 - ((3-pyridinyl) methyl) -1-azabicyclo [2.2.2] oct-3-yl) benzofuran-2-carboxamide p-toluenesulfonate

(2S, 3R) -N- (2 - ((3-pyridinyl) methyl) -1-azabicyclo [2.2.2] octan-3-yl) benzofuran-2-carboxamide (1.62 kg, 4.48 mol) and dichloromethane (8.60 kg) was added to the braided bottle. The percent weight / weight of the substance in the solution was determined using HPLC analysis. The solution was concentrated to an oil, acetone (4 L) was added and the mixture was concentrated to give an oily solid. Additionally, acetone (12 L) was added to the oily substance in the rotary evaporator flask, and the resulting suspension was transferred to a 50 L glass reaction flask with a mechanical stirrer, reflux condenser, temperature sensor and reflux condenser in an inert atmosphere. The reaction mixture was heated to 50 ° C ± 5 ° C. Water (80.7 g) was added to the solution and stirred for at least 10 minutes. P-Toluenesulfonic acid (853 g, 4.44 mol) was added portionwise to the reaction mixture over about 15 minutes. The reaction mixture was heated to boiling under reflux and kept at this temperature for at least 30 minutes to obtain a solution. The reaction mixture was cooled to 40 ° C ± 5 ° C for approximately 2 hours. Isopropyl acetate (14.1 L) was added over about 1.5 hours. The reaction mixture was slowly cooled to ambient temperature for at least 10 hours. The mixture was filtered and the filter cake washed with isopropyl acetate (3.5 L). The isolated product was dried in vacuum at 105 ° C ± 5 ° C for 2 to 9 hours, obtaining 2.19 kg (88.5% yield) of p-toluenesulfonate (2S, 3R) -N- (2 - ((3 -pyridinyl) methyl) -1-azabicyclo [2.2.2] oct-3-yl) benzofuran-2-carboxamide, so pl. 226-228 ° C. ¹ H NMR (500 MHz, D ₂ O) δ 8.29 (s, 1H), 7.78 (m, J = 5.1, 1H), 7.63 (d, J = 7.9, 1H) 7.54 (d, J = 7.8, 1H), 7.49 (d, J = 8.1, 2H), 7.37 (m, J = 8.3, 1H), 7.33 ( m, J = 8.3, 6.9, 1.0, 1H), 7.18 (m, J = 7.8, 6.9, 1.0, 1H), 7.14 (d, J = 8.1, 2H), 7.09 (s, 1H), 6.99 (dd, J = 7.9, 5.1, 1H), 4.05 (m, J = 7.7, 1H), 3.74 (m, 1H), 3.47 (m, 2H), 3.28 (m, 1H), 3.22 (m, 1H), 3.15 (dd, J = 13.2, 4, 7, 1H), 3.02 (dd, J = 13.2, 11.5, 1H), 2.19 (s, 3H), 2.02 (m, 2H), 1.93 (m, 2H) 1.79 (m, 1H). ¹³ C NMR (126 MHz, D ₂ O) δ 157.2, 154.1, 150.1, 148.2, 146.4, 145.2, 138.0, 137.0, 130.9, 128, 2 (2), 126.9, 126.8, 125.5 (2), 123.7, 123.3, 122.7, 111.7, 100.7, 61.3, 50.2, 48, 0, 40.9, 33.1, 26.9, 21.5, 20.8, 17.0.

Samples of this substance were converted to the free base of compound A (for use in salt selection studies) by treatment with an aqueous solution of sodium hydroxide and extraction with chloroform. As the chloroform was evaporated, an off-white powder remained, mp. 167-170 ° C, having the following spectral characteristics: electrospray in the mode of positive ions, mass spectrum [M + H] ⁺ ion m / z = 362. ¹ H NMR (500 MHz, DMSO-d ₆ ) δ 8.53 (d, J = 7.6 Hz, 1H), 8.43 (d, J = 1.7 Hz, 1H), 8.28 (dd , J = 1.6, 4.7 Hz, 1H), 7.77 (d, J = 7.7 Hz, 1H), 7.66 (d, J = 8.5 Hz, 1H), 7.63 (dt, J = 1.7, 7.7 Hz, 1H), 7.52 (s, 1H), 7.46 (m, J = 8.5, 7.5 Hz, 1H), 7.33 ( m, J = 7.7, 7.5 Hz, 1H), 7.21 (dd, J = 4.7, 7.7 Hz, 1H), 3.71 (m, J = 7.6 Hz, 1H ), 3.11 (m, 1H), 3.02 (m, 1H), 2.80 (m, 2H), 2.69 (m, 2H), 2.55 (m, 1H), 1.80 (m, 1H), 1.77 (m, 1H), 1.62 (m, 1H), 1.56 (m, 1H), 1.26 (m, 1H). ¹³ C NMR (126 MHz, DMSO-d ₆ ) δ 158.1, 154.1, 150.1, 149.1, 146.8, 136.4, 135.4, 127.1, 126.7, 123 6, 122.9, 122.6, 111.8, 109.3, 61.9, 53.4, 49.9, 40.3, 35.0, 28.1, 26.1, 19.6 .

The monohydrochloride salt of compound A (see Example 5) was subjected to X-ray diffraction crystallographic analysis. The resulting crystal structure (shown in FIGS. 10A and 10B, respectively) allows us to establish the 2S, 3R absolute configuration of compound A.

Example 3. Synthesis of the phosphate salt of (2S, 3R) -N- (2 - ((3-pyridinyl) methyl) -1-azabicyclo [2.2.2] oct-3-yl) benzofuran-2-carboxamide

(2S, 3R) -N- (2 - ((3-pyridinyl) methyl) -1-azabicyclo [2.2.2] oct-3-yl) benzofuran-2-carboxamide (8.18 g, 22) was added to the round bottom flask 6 mmol) and 2-propanol (180 ml). The mixture was stirred and heated at 65-70 ° C until all solids were dissolved. The solution was vigorously stirred at 65-70 ° C. and phosphoric acid (1.65 ml, 24.3 mmol) was added dropwise with a pipette. A white granular solid immediately formed. The mixture was stirred at 65-70 ° C for 30 minutes, cooled to ambient temperature (23 ° C) and further stirred for 24 hours. A white solid was collected by suction filtration by suction and the filter cake was washed with 2-propanol (20 ml) and the solid was dried in air for at least 1 hour. The solid was further dried in a vacuum oven at 75 ° C. overnight (16 hours) to obtain 10.7 g of product (> quantitative yield), mp 265-273 ° С with decomposition, with a change in crystallinity observed at ~ 180 ° С. The ¹ H NMR spectrum (DMSO-d ₆ ) showed the presence of 2-propanol (strong solvate), which may explain a yield exceeding quantitative. Chiral LC analysis showed a purity of 97.1% (270 nm).

Example 4. Synthesis of the maleate salt of (2S, 3R) -N- (2 - ((3-pyridinyl) methyl) -1-azabicyclo [2.2.2] oct-3-yl) benzofuran-2-carboxamide

Maleic acid (0.067 g, 0.630 mmol) was added to a hot suspension of (2S, 3R) -N- (2 - ((3-pyridinyl) methyl) -1-azabicyclo [2.2.2] oct-3-yl) benzofuran-2 -carboxamide (0.203 g, 0.560 mmol) in isopropyl acetate (2 ml), with the precipitation of a fine white solid together with a resinous precipitate. Additionally, isopropyl acetate (3 ml) and maleic acid (0.006 g) were added and the mixture was heated to boiling under reflux. Isopropanol (5 ml) was added under reflux. The resulting mixture of white solids was cooled to ambient temperature, filtered and the solids washed with isopropyl acetate (2 ml). The product was dried in vacuum at 60 ° C for 18 hours to obtain 0.228 g of an off-white flocculent solid (yield 84.7%), mp 180-182 ° C. The ¹ H NMR spectrum (DMSO-d ₆ ) indicated the stoichiometric composition of the monosalt. Calculated for C ₂₂ H ₂₃ N ₃ O ₂ ^. C ₄ H ₄ O ₄ : C, 65.40; H, 5.70; N, 8.80. Found: C, 65.35, 65.29; H, 5.86; 5.68; N, 8.69, 8.78.

Example 5. Synthesis of hydrochloride salts of (2S, 3R) -N- (2 - ((3-pyridinyl) methyl) -1-azabicyclo [2.2.2] oct-3-yl) benzofuran-2-carboxamide

Monohydrochloride: A solution of hydrochloric acid in THF was prepared by adding dropwise concentrated hydrochloric acid (1.93 ml of a 12M solution, 23.2 mmol) to 8.5 ml of chilled THF. The solution was heated to ambient temperature. (2S, 3R) -N- (2 - ((3-pyridinyl) methyl) -1-azabicyclo [2.2.2] oct-3-yl) benzofuran-2-carboxamide (8.49 g, 23) was added to a round bottom flask 5 mmol) and acetone (85 ml). The mixture was stirred and heated at 45-50 ° C until an absolute solution was obtained. The resulting yield. A solution of hydrochloric acid in THF was added dropwise over 5 minutes using an additional amount of THF (1.5 ml) for transfer. During the addition of the acid solution, a granular white solid began to form. The mixture was cooled to ambient temperature and stirred overnight (16 hours). The solid was collected by suction filtration under vacuum, the filter cake was washed with acetone (10 ml) and the solid was dried in air with suction under vacuum for 30 minutes. The solid was additionally dried in a vacuum oven at 75 ° C for 2 hours, obtaining 8.79 g of fine white crystals (yield 94%), so pl. 255-262 ° C. Chiral LC analysis gave a purity of 98.8% (270 nm). The ¹ H NMR spectrum (DMSO-d ₆ ) showed no residual solvents and confirmed a monostoichiometric composition. ¹ H NMR (300 MHz, DMSO-d ₆ ) δ 10.7 (broad s, 1H - quaternary ammonium), 8.80 (wide s, 1H - amide H), 8.54 (s, 1H), 8, 23 (d, 1H), 7.78 (d, 1H), 7.74 (d, 1H), 7.60 (d, 1H), 7.47 (m, 2H), 7.33 (m, 1H ), 7.19 (m, 1H), 4.19 (m, 1H), 4.08 (m, 1H), 3.05-3.55 (m, 6H), 2.00-2.10 ( m, 3H), 1.90 (m, 1H), 1.70 (m, 1H). X-ray crystallographic analysis of this salt established stereochemical assignment and stoichiometry (see figa and 10B).

Dihydrochloride: Hydrogen chloride gas was slowly bubbled into an ice-cooled solution of (2S, 3R) -N- (2 - ((3-pyridinyl) methyl) -1-azabicyclo [2.2.2] oct-3-yl) benzofuran-2-carboxamide (1.9 g, 5.3 mmol) in anhydrous ether (25 ml). Volatile components were removed, initially in a stream of nitrogen and then in high vacuum (a gas scrubber with sodium hydroxide was built into the high-vacuum line). The residue was triturated several times with small portions of anhydrous ether (discarded) and the remaining solid was dried in high vacuum. This gave 2.17 g (94% yield) of an off-white powder, mp. 210-212 ° C (hygroscopic). Chiral LC analysis gave a purity of 93.7% (270 nm). The mass spectrum by the method of electrospray in the mode of positive ions gave the ion [M + H] ⁺ m / z = 362. ¹ H NMR (300 MHz, CD ₃ OD) δ 9.15 (s, 1H), 8.84 (d, 1H), 8.63 (d, 1H), 7.97 (t, 1H), 7, 75 (d, 1H), 7.61 (d, 1H), 7.52 (m, 2H), 7.35 (t, 1H), 4.50 (m, 1H), 4.32 (m, 1H ), 3.40-3.85 (m, 6H), 1.95-2.40 (m, 5H).

Example 6. Synthesis of the hemigalactarate salt of (2S, 3R) -N- (2 - ((3-pyridinyl) methyl) -1-azabicyclo [2.2.2] oct-3-yl) benzofuran-2-carboxamide

Galactar (mucus) acid (36.3 mg, 0.173 mmol) was added to a solution of (2S, 3R) -N- (2 - ((3-pyridinyl) methyl) -1-azabicyclo [2.2.2] oct-3-yl ) benzofuran-2-carboxamide (125 mg, 0.346 mmol) in hot ethanol (1 ml). The mixture was heated at the boil under reflux after adding water (8 drops); then the hot mixture was filtered through a cotton plug, which was subsequently washed with ethanol (1 ml). Cooling did not precipitate. Volatile components were removed by evaporation using a rotary evaporator and the residue (white foam) was triturated with isopropanol (discarded) and the remaining solid was dissolved in an acetone / water mixture (4 ml in a 7: 1 ratio). Slow cooling to 5 ° C led to the formation of a white solid, which was filtered off, washed with isopropanol (3 × 1 ml) and dried in high vacuum. In the residue received 118 mg (yield 73%) of thin white plates, so pl. 134-139 ° C. ¹ H NMR (300 MHz, D ₂ O) δ 8.29 (s, 1H), 7.78 (d, 1H), 7.62 (d, 1H), 7.54 (d, 1H), 7, 35 (m, 2H), 7.18 (t, 1H), 7.10 (s, 1H), 6.98 (m, 1H), 4.08 (s, 1H, galactaric acid), 3.98 ( d, 1H), 3.77 (s, 1H, galactaric acid), 3.66 (m, 1H), 3.35 (m, 1H), 2.95-3.30 (m, 4H), 1, 65-2.05 (m, 5H).

Example 7. Synthesis of the D-tartrate salt of (2S, 3R) -N- (2 - ((3-pyridinyl) methyl) -1-azabicyclo [2.2.2] oct-3-yl) benzofuran-2-carboxamide

Tartaric acid (25.6 mg, 0.173 mmol) was added (2S, 3R) -N- (2 - ((3-pyridinyl) methyl) -1-azabicyclo [2.2.2] oct-3-yl) benzofuran-2- carboxamide (125 mg, 0.346 mmol) in hot ethanol (1 ml). The resulting solution was slowly cooled to ambient temperature. The solid did not precipitate, so the solution was concentrated to give a white foam. Attempts to crystallize in isopropanol were unsuccessful. The foam was dissolved in methanol and a second portion of half the equivalent of tartaric acid (25.6 mg, 0.173 mmol) was added. The mixture was concentrated to give a white foam that did not crystallize from mixtures of methanol and isopropanol. The concentrated substance (a mixture of a solid and a gummy liquid) was then suspended in ethyl acetate (1 ml) to obtain a white solid. It was isolated by filtration (washed with ethyl acetate) and dried in a vacuum oven (18 hours at 40 ° C) to obtain 141 mg (79.7% yield) of salt with a monostoichiometric composition (NMR), mp 136-140 ° C. Chiral LC analysis gave 98.1% purity (270 nm). ¹ H NMR (300 MHz, D ₂ O) δ 8.50 (s, 1H), 8.01 (d, 1H), 7.86 (d, 1H), 7.75 (d, 1H), 7, 56 (m, 2H), 7.38 (t, 1H), 7.32 (s, 1H), 7.21 (t, 1H), 4.34 (s, 2H, tartaric acid), 4.26 ( d, 1H), 3.95 (m, 1H), 3.64 (m, 2H), 3.15-3.55 (m, 4H), 1.90-2.30 (m, 5H).

Example 8. Synthesis of the methanesulfonate salt of (2S, 3R) -N- (2 - ((3-pyridinyl) methyl) -1-azabicyclo [2.2.2] oct-3-yl) benzofuran-2-carboxamide

Methanesulfonic acid (33.2 mg, 0.346 mmol) was added to a solution of (2S, 3R) -N- (2 - ((3-pyridinyl) methyl) -1-azabicyclo [2.2.2] oct-3-yl) benzofuran- 2-carboxamide (125 mg, 0.346 mmol) in hot ethanol (1 ml). Cooling led to the formation of a precipitate. The mixture was heated to boiling under reflux and the hot mixture was filtered through a cotton plug, which was subsequently washed with methanol (1 ml). Volatile components were removed using a rotary evaporator and the residue (light yellow foam) was dissolved in hot isopropanol (1 ml). Again, cooling led to the formation of a precipitate. Isopropanol was evaporated and the residue was suspended in acetone (1 ml). Filtration and drying in a vacuum oven (18 hours at 50 ° C) gave 146 mg (yield 92.5%) of a light beige solid, mp. 240-243 ° C. ¹ H NMR (300 MHz, D ₂ O) δ 8.32 (s, 1H), 7.82 (d, 1H), 7.66 (d, 1H), 7.57 (d, 1H), 7, 38 (m, 2H), 7.20 (m, 1H), 7.12 (s, 1H), 7.01 (m, 1H), 4.09 (d, 1H), 3.75 (m, 1H ), 3.47 (m, 2H), 3.00-3.40 (m, 4H), 2.67 (s, 3H, methanesulfonic acid), 1.75-2.15 (m, 5H).

Example 9. Synthesis of the D-mandelate salt of (2S, 3R) -N- (2 - ((3-pyridinyl) methyl) -1-azabicyclo [2.2.2] oct-3-yl) benzofuran-2-carboxamide

D-mandelic acid (52.6 mg, 0.346 mmol) was added to a solution of (2S, 3R) -N- (2 - ((3-pyridinyl) methyl) -1-azabicyclo [2.2.2] oct-3-yl) benzofuran-2-carboxamide (125 mg, 0.346 mmol) in hot ethanol (1 ml). Dilution with ethyl acetate (4 ml) and cooling did not precipitate. Volatile components were removed by evaporation using a rotary evaporator and the residue (white foam) was dissolved in hot isopropanol (0.5 ml). Cooling to 5 ° C led to the formation of white crystals, which were collected by suction filtration in vacuo. Drying in a vacuum oven (18 hours at 45 ° C) gave 111 mg (yield 62.4%) of a light beige solid, mp. 188.5-193 ° C. ¹ H NMR (300 MHz, D ₂ O) δ 8.33 (s, 1H), 7.83 (s, 1H), 7.67 (d, 1H), 7.60 (d, 1H), 7, 27 (m, 8H, includes mandelic acid), 7.12 (s, 1H), 7.01 (m, 1H), 4.85 (s, 1H, mandelic acid), 4.10 (d, 1H), 3.75 (m, 1H), 3.48 (m, 2H), 3.00-3.40 (m, 4H), 1.75-2.15 (m, 5H).

Example 10. Synthesis of the R-camphorsulfonate salt of (2S, 3R) -N- (2 - ((3-pyridinyl) methyl) -1-azabicyclo [2.2.2] oct-3-yl) benzofuran-2-carboxamide

R-10-Camphorsulfonic acid (80.3 mg, 0.346 mmol) was added to a solution of (2S, 3R) -N- (2 - ((3-pyridinyl) methyl) -1-azabicyclo [2.2.2] oct-3- il) benzofuran-2-carboxamide (125 mg, 0.346 mmol) in hot ethanol (1 ml). Cooling did not precipitate any precipitate. Volatile components were removed by evaporation using a rotary evaporator and the residue (white foam) was dissolved in hot isopropanol (0.5 ml). Cooling to 5 ° C led to the formation of individual white crystals and milk suspension. Scratching the walls of the flask with a spatula over time led to the conversion of the mixture into a dense mass of small white crystals. An additional 0.5 ml of isopropanol was added and the crystals were collected by filtration with suction of the precipitate in vacuo. Drying in a vacuum oven (5 hours at 70 ° C, then 2 hours at 110 ° C) gave 193 mg (yield 93.8%) of a white solid, mp 149,5-156 ^{C. 1} H NMR (300 MHz, D ₂ O) δ 8,30 (s, 1H), 7.79 (d, 1H), 7.64 (d, 1H), 7.55 (d, 1H), 7.36 (m, 2H), 7.18 (m, 1H), 7.11 (s, 1H), 6.99 (m, 1H), 4.07 (d, 1H) 3.73 (m, 1H), 3.45 (m, 2H), 3.95-3.35 (m, 5H, includes camphorsulfonic acid), 2.64 (d, 1H, camphorsulfonic acid), 2, 22 (m, 2H), 1.70-2.10 (m, 8H, includes camphorsulfonic acid), 1.45 (m, 1H, camphorsulfonic acid), 1.25 (m, 1H, camphorsulfonic acid), 0, 85 (s, 3H, camphorsulfonic acid); 0.68 (s, 3H, camphorsulfonic acid).

Example 11. Synthesis of the S-camphorsulfonate salt of (2S, 3R) -N- (2 - ((3-pyridinyl) methyl) -1-azabicyclo [2.2.2] oct-3-yl) benzofuran-2-carboxamide

S-10-Camphorsulfonic acid (80.3 mg, 0.346 mmol) was added to a solution of (2S, 3R) -N- (2 - ((3-pyridinyl) methyl) -1-azabicyclo [2.2.2] oct-3- il) benzofuran-2-carboxamide (125 mg, 0.346 mmol) in hot ethanol (1 ml). Dilution with ethyl acetate (4 ml) and cooling did not precipitate any precipitate. Volatile components were removed by evaporation using a rotary evaporator and the residue (white foam) was dissolved in hot isopropanol (1.5 ml). Cooling to 5 ° C led to the formation of white crystals. The mixture was concentrated to ~ 0.5 ml and again cooled to 5 ° C. The solid was then collected by suction filtration by suction and dried in vacuo, initially 18 hours at 45 ° C, but then gradually at a higher temperature (finally at 110 ° C) to remove residual isopropanol. This gave 143 mg (yield 69.7%) of a white solid, mp. 153.5-157 ° C. ¹ H NMR (300 MHz, D ₂ O) δ 8.29 (s, 1H), 7.79 (d, 1H), 7.63 (d, 1H), 7.54 (d, 1H), 7, 34 (m, 2H), 7.18 (m, 1H), 7.10 (s, 1H), 6.99 (m, 1H), 4.05 (d, 1H), 3.73 (m, 1H ), 3.44 (m, 2H), 3.95-3.35 (m, 5H, includes camphorsulfonic acid), 2.67 (d, 1H, camphorsulfonic acid), 2.23 (m, 2H), 1 70-2.10 (m, 8H, includes camphorsulfonic acid), 1.46 (m, 1H, camphorsulfonic acid), 1.25 (m, 1H, camphorsulfonic acid), 0.84 (s, 3H, camphorsulfonic acid ), 0.64 (s, 3H, camphorsulfonic acid).

Using methods similar to those described above (examples 3-11), several other salt forms were characterized. The results of obtaining such compounds are shown in examples 12-14.

Example 12. Synthesis of the sulfate salt of (2S, 3R) -N- (2 - ((3-pyridinyl) methyl) -1-azabicyclo [2.2.2] oct-3-yl) benzofuran-2-carboxamide

The sulfate salt was precipitated from a mixture of isopropyl acetate and water. Mp 278 ° C. ¹ H NMR (400 MHz, DMSO-d ₆ ) δ 9.28 (broad s, 1H, amide), 8.56 (dd, 1H), 8.24 (t, 1H), 7.77 (d, 1H) ), 7.74 (d, 1H), 7.60 (s, 1H), 7.40 (m, 2H), 7.35 (s, 1H), 7.21 (m, 1H), 4.21 (m, 1H), 3.93 (m, 2H), 3.10-3.60 (m, 5H), 2.05 (m, 3H), 1.92 (m, 1H), 1.73 ( m, 1H).

Example 13. Synthesis of the ketoglutarate salt of (2S, 3R) -N- (2 - ((3-pyridinyl) methyl) -1-azabicyclo [2.2.2] oct-3-yl) benzofuran-2-carboxamide

The α-ketoglutarate salt was precipitated from isopropyl acetate. Mp 177 ° C. ¹ H NMR (400 MHz, DMSO-d ₆ ) δ 8.64 (s, 1H, amide), 8.50 (d, 1H), 8.20 (d, 1H), 7.74 (d, 1H) , 7.70 (d, 1H), 7.60 (m, 1H), 7.45 (m, 1H), 7.32 (m, 2H), 7.18 (m, 1H), 4.10 ( m, 1H), 3.78 (m, 2H), 3.00-3.45 (m, 5H), 2.81 (m, 2H, ketoglutaric acid), 2.41 (m, 2H, ketoglutaric acid) 1.96 (m, 3H); 1.83 (m, 1H); 1.60 (m, 1H).

Example 14. Synthesis of the hippurate salt of (2S, 3R) -N- (2 - ((3-pyridinyl) methyl) -1-azabicyclo [2.2.2] oct-3-yl) benzofuran-2-carboxamide

The hippurate salt was precipitated from acetone (too hygroscopic to obtain a melting point). ¹ H NMR (400 MHz, DMSO-d ₆ ) δ 8.79 (s, 1H, amide), 8.56 (d, 1H), 8.44 (s, 1H, hippuric acid), 8.29 (m , 1H), 7.87 (m, 2H, hippuric acid), 7.76 (d, 1H), 7.65 (m, 1H), 7.54 (m, 1H), 7.49 (m, 4H) includes hippuric acid), 7.34 (m, 2H), 7.21 (m, 1H), 3.91 (m, 1H), 3.74 (m, 2H), 3.00-3.50 ( m, 5H), 2.80 (m, 2H, hippuric acid), 1.79 (m, 2H), 1.60 (m, 2H), 1.30 (m, 1H).

Example 15. Isolation of (2R, 3R) - and (2S, 3S) -N- (2 - ((3-pyridinyl) methyl) -1-azabicyclo [2.2.2] oct-3-yl) benzofuran-2-carboxamide and conversion to galactaric acid salts

A sample of the supernatant from the experiment on the isolation of (2S, 3R) -N- (2 - ((3-pyridinyl) methyl) -1-azabicyclo [2.2.2] oct-3-yl) benzofuran-2-carboxamide p-toluenesulfonate ( Example 2) was concentrated by evaporation on a rotary evaporator, adjusted to pH 10 with 10% aqueous sodium hydroxide and extracted with dichloromethane. The dichloromethane extract was evaporated and the residue (1.8 g) was dissolved in absolute ethanol (55 ml) containing 0.5% di-n-butylamine. This solution was injected in 0.25 ml portions per 25 cm × 2.1 cm Chiralpak® AD column by chiral HPLC, eluting with a 60:40: 0.2 mixture of hexane / ethanol / di-n-butylamine (flow rate = 30 ml / min ) by monitoring at 270 nm. Isolation of the effluent eluted at ~ 7.5 minutes and then elution at ~ 13.5 minutes gave, after evaporation of the solvent, 0.48 g (98% chiral purity) and 0.47 g (99% chiral purity), respectively, of a colorless oil. The two NMR spectra were the same. ¹ H NMR (300 MHz, CDCl ₃ ) δ 8.49 (s, 1H), 8.45 (d, 1H), 7.74 (d, 1H), 7.52 (m, 4H), 7.35 (t, 1H), 7.20 (dd, 1H), 7.05 (d, 1H), 4.55 (dt, 1H), 3.43 (m, 1H), 3.22 (m, 1H) 2.90 (m, 5H); 2.09 (m, 1H); 1.88 (m, 4H).

A warm solution of each of the samples of the free base in absolute ethanol (10 ml) was treated with one equivalent of galactaric acid. The resulting mixture was heated at 75 ° C for 5 minutes and cooled with stirring to ambient temperature. The resulting solids were collected by suction filtration the precipitate in vacuo and dried in vacuo to give 0.65 g (yield 87%) and 0.62 g (yield 85%), respectively, a white granular solid (m.p. 200-205 ^{° C} in each case). ¹ H NMR (300 MHz, D ₂ O) δ 8.38 (s, 1H), 8.28 (d, 1H), 7.94 (d, 1H), 7.70 (d, 1H), 7, 59 (d, 1H), 7.48 (t, 1H), 7.40 (m, 1H), 7.32 (m, 2H), 4.42 (m, 1H), 4.21 (s, 2H) ), 3.87 (s, 2H), 3.68 (m, 1H), 3.35 (m, 6H), 2.25 (m, 2H), 2.02 (m, 3H).

Example 16. Synthesis of the p-chlorobenzoate salt of (2R, 3R) -N- (2 - ((3-pyridinyl) methyl) -1-azabicyclo [2.2.2] oct-3-yl) benzofuran-2-carboxamide

Solid p-chlorobenzoic acid (46.8 mg, 0.299 mmol) was added in one portion to a solution of the previously isolated N- (2 - ((3-pyridinyl) methyl) -1-azabicyclo [2.2.2] oct-3- isomer il) benzofuran-2-carboxamide from Example 15 (108 mg, 0.299 mmol) in acetone (10 ml). This mixture was heated almost at the boil under reflux for 30 minutes and cooled to ambient temperature. A precipitate did not form, so the solution was concentrated to about 20% of its original volume (hot cup), at which point crystals began to form. The mixture was cooled and diluted with isopropanol (2 ml).

This mixture was concentrated by slowly evaporating the solvent at ambient temperature and the resulting solid was collected and dried. This product, 145 mg (yield 94%) was a light yellow crystals, so pl. 150-152 ° C. ¹ H NMR (300 MHz, CDCl ₃ ) δ 8.49 (s, 1H), 8.38 (d, 1H), 7.93 (d, 2H, p-chlorobenzoic acid), 7.67 (m, 2H ), 7.57 (d, 1H), 7.45 (m, 1H), 7.36 (d, 2H, p-chlorobenzoic acid), 7.30 (m, 1H), 7.27 (s, 1H ), 7.16 (m, 1H), 7.00 (d, 1H, amide), 6.90 (broad s, quaternary ammonium), 4.62 (m, 1H), 3.85 (dd, 1H) 3.36 (m, 1H); 2.95-3.25 (m, 5H); 2.16 (s, 1H); 1.70-2.10 (m, 4H).

According to x-ray diffraction studies of this sample, its absolute stereochemistry 2R, 3R (see figa and 11B). Accordingly, by way of exception, the last eluted isomer in Example 15 has the absolute configuration 2S, 3S.

Example 17. Chiral chromatographic method for the analysis of stereoisomers

The creation of a chiral chromatographic method for the separation of four stereoisomers, one from the other, seemed very promising. Initial attempts (using the hexane / isopropanol / triethylamine mobile phase) resulted in overlapping peaks and less than optimal peak shapes. The transition from isopropanol to ethanol and from triethylamine to di-n-butylamine improved separation and peak shape and reduced transit time. The details of the method are as follows:

Analytical column: Chiralpak® AD (250x4.6 mm, 5 μm)

Mobile phase: 60: 40: 0.2 hexanes / ethanol / di-n-butylamine

Injection volume: 10 μl

Flow rate: 1.0 ml per minute

Temperature: 20 ° C

Detection: UV at 270 nm

Total passage time: ~ 25 minutes

The elution order (retention time): 2S, 3R (5.3 min); 2R, 3S (7.3 min); 2R3R (8.3 min); 2S, 3S (12.1 min)

An exemplary chromatogram of stereoisomeric analogs is shown in FIG.

Example 18. X-ray powder diffraction analysis (XRPD)

X-ray powder diffraction analysis (XRPD) was performed for several samples of the salts described herein. The nature of the distribution of peaks for the hydrochloride (Fig.13) and tosylate (Fig.14) salts is given.

X-ray powder diffraction analysis (XRPD)

Peaks in the X-ray powder diffraction pattern were obtained on each of or on both of the two devices. In some cases, X-ray diffraction patterns were obtained on a Siemens D5000 diffractometer using CuKα radiation (40 kV, 40 mA), a θ-θ goniometer, V20 divergence and receiving slits, a graphite secondary monochromator and a scintillation counter. Instrument operation was verified using a certified corundum standard (NIST 1976). The passage of samples under ambient conditions was carried out on a flat plate for samples using powders in the form in which they were obtained. Approximately 35 mg of the sample was carefully placed in a cavity cut into a polished silicone substrate with zero background (510). The sample was rotated in its own plane during analysis, scanning from 2 ° to 42 ° in increments of 0.05 °, at the rate of 4 seconds per step, using CuKα1 (λ = 1.5406 Å).

In a number of cases, X-ray diffraction patterns were obtained on a Bruker AXS C2 GADDS diffractometer using CuKα radiation (40 kV, 40 mA), an automated XYZ stage, a laser video microscope for self-feeding a sample, and a 2-dimensional HiStar area detector. The diffractometer optics consists of one multilayer Goebel mirror connected to a collimator with a point aperture of 0.3 mm. The beam divergence (i.e., the effective x-ray beam size on the sample) was approximately 4 mm. A continuous θ-θ scanning mode was used with a sample-detector distance of 20 cm, which gave an effective 2θ range of 3.2 ° -30.0 °. Typically, the sample was exposed to an x-ray for 120 seconds. The passage of samples under ambient conditions was carried out on a flat plate for samples using powders in the form in which they were obtained without grinding. Approximately 1-2 mg of the sample was lightly pressed onto a silicone substrate to obtain a flat surface. Samples for conducting the experiment under non-environmental conditions were mounted on a silicone substrate using a heat-conducting compound. The sample was then heated to a suitable temperature at a rate of about 10 ° C / min and then kept under isothermal conditions for about 5 minutes before starting data collection.

Differential Scanning Calorimetry (DSC)

DSC data was obtained on TA Q1000 instruments equipped with an automatic device for feeding a sample at 50 positions. The instrument has been calibrated to calibrate energy and temperature using certified indium. Typically 0.5-1.5 mg of each sample in an aluminum cup with a pinhole was heated at 10 ° ^C / min from 25 ° C to 175-200 ° C. A stream of nitrogen was maintained above the samples at a rate of 30 ml / min.

Thermogravimetric Analysis (TGA)

TGA data was obtained on a TA Q500 (TGA) instrument equipped with an automatic device for feeding a sample at 16 positions. The instrument was temperature calibrated using a certified alumel. Typically, 5-10 mg of each sample was loaded into a pre-weighed platinum crucible and an aluminum DSC dish and heated at a rate of 10 ° C / min from ambient temperature to 350 ° C. A flow of nitrogen was maintained over the samples at a rate of 60 ml / min.

Polarized Light Microscopy (MPS)

Samples were examined in polarized light on a Leica LM / DM microscope with a digital video camera for image capture. A small amount of each sample was placed on a coverslip in immersion oil and covered with a coverslip, individual particles were separated from each other as much as possible. Samples were viewed at a suitable magnification in partially polarized light coupled to a λ spectrozonal filter.

High temperature microscopy

High temperature microscopy was performed using a Leica LM / DM microscope in polarized light coupled to a Mettler-Toledo MTFP82HT hot plate and a digital video camera for image capture. A small amount of each sample was placed on a coverslip and individual particles were separated from each other as much as possible. Samples were viewed at a suitable magnification in partially polarized light coupled to a λ spectrozonal filter as it was heated from ambient temperature at a rate of usually 10 ° C / min.

Gravimetric sorption of steam (GSP)

Sorption isotherms were determined on either of two or both instruments. A series of experiments were performed using a VTI Corporation SGA-100 moisture sorption analyzer controlled by VTI FlowSystem 4 software. The sample temperature was maintained at 25 ° C using a constant thermostatic bath Polyscience. Humidity was controlled by mixing flows of dry and wet nitrogen. Weight changes as a function of% relative humidity (RH) were monitored using a Cahn Digital Recording Balance D200 with an accuracy of +/- 0.0001 g.

Typically, 10-20 mg samples were placed on a balanced weighing dish under ambient conditions. The sample was dried at 50 ° C for 1 hour. The standard adsorption isotherm was obtained at 25 ° C with 5% RH intervals in the range of 5-95% RH and the desorption isotherm was similarly obtained at 25 ° C with 5% RH intervals in the range of 95-5% RH. The criterion for balancing the sample included 0.0100% weight. for 5 minutes at a maximum equilibrium time of 180 minutes for each point% OV.

Some sorption isotherms were obtained using a Hiden IGASorp sorption moisture analyzer controlled by CFRSorp software. The temperature of the sample was maintained at 25 ° C. using a Huber recirculating water bath. Humidity was controlled by mixing dry and wet nitrogen streams at a total flow rate of 250 ml / min. Relative humidity was measured using a calibrated Vaisala RH probe (dynamic range 0-95% RH), located next to the sample. Changes in weight, (mass relaxation) of the sample as a function of% RH were constantly monitored by microbalance (accuracy ± 0.001 mg). Typically, 10-20 mg of the sample was placed in a balanced basket with stainless steel cells at ambient conditions. The sample was loaded and unloaded at 40% RH and 25 ° C (typical environmental conditions). The moisture sorption isotherm was obtained as briefly described below (2 scans gave 1 full cycle). The standard isotherm was obtained at 25 ° C at intervals of 10% RH in the range of 0-90% RH.

Generalized parameters of the SHG method

Options Values Adsorption - Scan 1 40-90 Desorption / Adsorption - Scan 2 85 - dry, dry - 40 Intervals (% RH) 10 The number of scans 2 Flow rate (ml / min) 250 Temperature (° C) 25 Stability (° C / min) 0.05 Minimum sorption time (hours) one Maximum sorption time (hours) four Mode AF2 Accuracy (%) 98

The software used the least squares minimization method together with the mass relaxation model. The measured value of mass relaxation should be within 5% of that predicted using software before choosing the next value in% RH. The minimum equilibration time was set at 1 hour, and the maximum at 4 hours. Typically, samples after completion of isotherm production were reanalyzed by X-ray powder diffraction (XRPD) analysis.

Karl Fischer definition of water

The water content in each sample was measured on a Mettler Toledo DL39 coulometer using a Hydranal Coulomat AG reagent and an argon pump. Weighed solid samples were introduced into a vessel on a platinum plate for TGA, which was connected to a rubber septum to prevent access of water. About 10 mg titration samples were used, and duplicate determination was performed.

Determination of thermodynamic water solubility by HPLC

Water solubility was determined by suspending a sufficient amount of the compound in 0.25 ml of water to obtain a maximum final concentration of ≥10 mg / ml of the original free form of the compound. The suspension was balanced at 25 ° C. for 24 hours and then the pH was measured. The suspension was filtered through a glass porous filter C into 96-well plates. The filtrate was then diluted with factor 101. Quantification was performed using HPLC with reference to a standard solution with a concentration of approximately 0.1 mg / ml in DMSO. Sprayed standard, diluted and undiluted sample solution. Solubility was calculated using peak areas determined by integrating the peak detected at the same retention time as the main peak upon injection of the standard. If there was enough solid substance on the filter cup, then powder diffractometry (XRPD) data was obtained

General details of a method for determining thermodynamic water solubility

Type of method Reverse phase with gradient elution Speaker Phenomenex Luna, C18 (2) 5 μm,
50 × 4.6 mm Column temperature ( ^о С): 25 Injection (μl): 5, 8 and 50 Detection:
Wavelength, bandwidth (nm): 260, 80 Flow rate (ml / min): 2 Phase A: 0.1% TFA in water Phase B: 0.085% TFA in acetonitrile

Timing chart Time (min) % Phase A % Phase B 0,0 95 5 1,0 80 twenty 2,3 5 95 3.3 5 95 3,5 95 5 4.4 95 5

Determination of chemical purity by HPLC

Purity analysis was performed on an Agilent HP1100 series system equipped with a diode array detector and using ChemStation software. Used one of two methods, described in detail below.

Method 1

Type of method Reverse phase with gradient elution Speaker Kromasil 5 μm C18, 150 × 4.6 mm Column temperature (° C): 26 Injection (μl): 10 Detection:
Wavelength, bandwidth (nm): 302, 8 Flow rate (ml / min): 1,0 Phase A: 0.0256 M KH2PO4 + 0.02 M Na 1-hexanesulfonic acid salt Phase B: acetonitrile Timing chart Time (min) % Phase A % Phase B 0 90 10 8 90 10 40 10 90 41 90 10 49 90 10 fifty 90 10

Method 2

Type of method Reverse phase with gradient elution Speaker Phenomenex Luna, C18 (2), 150 × 4.6 mm, 5 μm, Column temperature (° C): 25 Injection (μl): 5 Detection:
Wavelength, bandwidth (nm): 255.90 Flow rate (ml / min): one Phase A: 0.1% TFA in water Phase B: 0.085% TFA in acetonitrile Timing chart Time (min) % Phase A % Phase B 0 95 5 25 5 95 25,2 95 5 thirty 95 5

Ion chromatography

Data was obtained on a Metrohm 861 Advanced Compact IC instrument using IC Net software, version 2.3. Samples were obtained as stock solutions at a concentration of 1000 ppm (ppm) in water. When the solubility of the samples was low, a suitable cosolvent such as DMSO was used. Samples were diluted to a concentration of 50 ppm or 100 ppm using a suitable solvent before testing. Quantification was carried out by comparison with standard solutions with known concentrations of the analyzed ion.

Ion chromatography method for anions

Type of method Anion exchange Speaker Metrosep A Supp 5-250 (4.0 × 250 mm) Column temperature ( ^о С): Ambient temperature Injection (μl): twenty Detection: Conductivity detector Flow rate (ml / min): 0.7 Eluent 3.2 mM sodium carbonate
1.0 mM sodium bicarbonate in water

Ion chromatography method for cations

Type of method Cation exchange Speaker Metrosep C 2-250 (4.0 × 250 mm) Column temperature (° C): Ambient temperature Injection (μl): twenty Detection: Conductivity detector Flow rate (ml / min): 1,0 Eluent 4.0 mM tartaric acid
0.75 mM dipicolinic acid in water

Approximately 50 mg of (2S, 3R) -N- (2 - ((3-pyridinyl) methyl) -1-azabicyclo [2.2.2] oct-3-yl) benzofuran-2-carboxamide hydrochloride was weighed in a glass vial and heated to 50 ° C. Portions of 100 μl of 1-butanol / water mixture (5% water by volume) were added to the solid until a clear solution was formed (500 μl total). The sample was stirred at 50 ° C. for 1 hour and observed. After heating at 50 ° C for an hour, the sample continued to remain a clear solution and was cooled from 50 ° C to 25 ° C at a rate of 1.4 ° C per hour. The sample remained a clear solution upon cooling, and it was covered with parafilm with a pin hole and allowed to evaporate at ambient temperature. After 2 weeks, large crystals were visible in the partially stripped off sample. 13 shows a powder X-ray diffraction pattern (XRPD) of (2S, 3R) -N- (2 - ((3-pyridinyl) methyl) -1-azabicyclo [2.2.2] oct-3-yl) benzofuran-2-carboxamide monohydrochloride , illustrating both the observed (lighter) and calculated (darker) peaks.

The experimental peak distribution was obtained for a sample of (2S, 3R) -N- (2 - ((3-pyridinyl) methyl) -1-azabicyclo [2.2.2] oct-3-yl) benzofuran-2-carboxamide monohydrochloride, whereas calculated examples of peaks were obtained from an X-ray diffraction experiment for a single crystal, as described herein and shown in FIGS. 10A and 10B. Both sets of peaks are consistent with respect to 2θ values, and slight differences in peak intensities and widths may be related to instrument resolution and preferred orientation effects. In addition, minimal differences can be associated with the temperature shift, since the observed data were obtained at room temperature, and the calculation data were obtained for the structure at 120 K.

The structure of the tosylate salt, in particular, the crystalline monosalt, and the nature of the distribution of the peaks in the diffractogram of FIG. 14 were confirmed, the data were obtained using CuKα radiation (40 kV, 40 mA), θ-θ goniometer, V20 divergence and receiving slits, secondary graphite monochromator and scintillation counter. The XRPD diffraction pattern of the tosylate salt after 1 week of storage at 40 ° C / 75% relative humidity revealed changes, but the sample is still in Form 1. Probably, the changes are associated with a more hydrated form.

VII. Biological tests

The ability of compound A and its stereoisomers to bind to various NNR subtypes and modulate their function has been evaluated as described in US Pat. The assessment of the selectivity profile for receptors for compound A (including 5HT ₃ and muscarinic) was performed using NovaScreen® Bioscience Corporation.

Electrophysiological measurements of the α7 NNR response were performed in two expression systems: rat α7 NNR in mammalian GH4C1 cells and human α7 NNR in Xenopus oocytes.

GH4C1 cells expressing rat α7 NNRs were obtained as described in Placzek et al., Mol. Pharm. 68 (6): 1863-1876 (2005), incorporated herein by reference. Electrophysiological measurements of agonistic activity were performed using the Dynaflow rapid perfusion system and the patch clamp method using the GH4C1 cell expression system. Both acetylcholine and nicotine induced concentration-dependent activation of the α7-mediated current. Agonistic EC ₅₀ values from the literature were comparable to those obtained by this method (see, Biochem Pharmacol, in print (2007) and available online Dynaflow materials (www.cellectricon.com), both of which are incorporated into this description by reference to this way).

Whole cell currents recorded using an Axopatch 700A amplifier were filtered at 1 kHz and tested at 5 kHz using a PCI card (National Instrument). Compared with previous studies, saline solutions were modified as indicated to increase current stability. Cells were recorded at room temperature in the following intercellular medium: 130 mM NaCl, 5 mM KCl, 2 mM CaCl ₂ , 2 mM MgCl ₂ , 10 mM HEPES adjusted to pH 7.4 with aqueous NaOH. Borosilicate electrodes (3-5 MΩ) were filled with the following medium: 130 mM Tris-phosphate, 5 mM NaCl, 2 mM MgCl ₂ , 10 mm HEPES, 10 mM EGTA adjusted to pH 7.4 with aqueous KOH (see Wu et al. , J. Physiol. 576: 103-118 (2006), incorporated herein by reference to describe such indications). Under these conditions, macrocurrent activity obtained using NNR registration in whole cells lasts up to 60 minutes when it is caused by a concentration of acetylcholine (ACh) of 1000 μM.

The methods of working with cells have been adapted according to the guidelines for the use of Cellectricon for Dynaflow. Briefly, after removal from the incubator, the cells were thoroughly washed three times with recording medium and placed on a Zeiss microscope stand. An average of 5 minutes was required before establishing the registration configuration for whole cells. To exclude modification of cellular conditions, one cell was registered for a load of cells in a Dynaflow silicone chip. For experimental conditions, differences in the fraction of cells that give a response could not be detected. More than 95% of the cells responded to Ach, and each cell present in the measured current was taken into account. Cells were maintained at -60 mV for the duration of the experiment. All test solutions for work were made daily from stock solutions. A fresh stock solution of acetylcholine (ACh) was prepared daily in Ringer's solution and diluted. Dose response curves were described by a single Hill equation using Prism 5.0 software.

Xenopus oocytes expressing human α7 NNRs were prepared as described by Papke and Papke, Brit. J. Pharmacol. 137: 49-61 (2002), incorporated herein by reference. Mature females (> 9 cm) of African toads Xenopus laevis (Nasco, Ft. Atkinson, WI) were used as a source of oocytes. Before surgery, toads were anesthetized by placing the animal in a solution of 1.5 g / L 3-aminobenzoic acid ethyl ester for 30 minutes. Oocytes were removed through an abdominal incision.

To remove the follicular cell layer, the collected oocytes were treated with 1.25 mg / ml collagenase from Worthington Biochemical Corporation (Freehold, NJ) for 2 hours at room temperature in a calcium-free Bart solution (88 mM NaCl, 1 mM KCl, 15 mM HEPES, pH 7.6, 0.81 mM MgSO ₄ , 2.38 mM NaHCO ₃ , 0.1 mg / ml gentamicin sulfate). Then stage 5 oocytes were isolated and 50 nl (5-20 ng) of human cRNA α7 was injected into each injection. Registration was performed 2-7 days after injection. Fresh stock solutions of acetylcholine (ACh) were prepared daily in Ringer's solution.

The experiments were performed using OpusXpress 6000A (Axon Instruments, Union City, CA). OpusXpress is an integrated system that provides automatic insertion and fixation of stress in parallel for eight oocytes. Electrodes of both voltage and current filled 3M KCl. For cells, the voltage was fixed at a holding potential of -60 mV. Data was obtained at 50 Hz and filtered at 20 Hz. Cells were poured into the bath with Ringer's solution and agonist solutions were delivered from a 96-well plate using disposable tips, which excluded any possibility of cross-contamination. Flow rates were set at 2 ml / min. The use of the drug was alternated between ACh controls and experimental agonists. The use of the product was 12 seconds in duration, followed by a washing period of 181 seconds.

Responses were calculated as the net charge (see Papke and Papke, Brit. J. Pharmacol. 137: 49-61 (2002), as cited above) for α7 receptors. For each oocyte, the initial control application of ACh was performed, then the experimental drug was applied, and then the subsequent control application of ACh (300 μM) was performed. Responses to the application of the experimental drug were calculated relative to the previous control response to ACh to normalize the data, compensating for changes in the levels of channel expression among oocytes. Note that 300 μM ACh caused maximal responses of the resulting charge from α7 receptors, so that normalization to ACh controls effectively normalized the data to maximal responses to ACh. Mean values and standard errors (SEM) were calculated from normalized responses for at least four oocytes for each experimental concentration. For the concentration-response ratios, for the data obtained from the analysis of the resulting charges, a graph was constructed using the Kaleidagraph 3.0.2 software (Abelbeck software, Reading, PA) and curves were generated from the Hill equation.

Behavioral characterization of compound A was carried out in accordance with the following protocols. The object recognition task was carried out in accordance with the description of Ennaceur and Delacour, Behav. Brain Res. 100: 85-92 (1988), incorporated herein by reference. The angle-arm labyrinth paradigm (RAM) was performed as described by Levin et al., Behav. Pharm. 10: 675-680 (1999), incorporated herein by reference. A prepulse inhibition assay (PPI) was performed as described by Suemaru et al., Brit. J. Pharmocol. 142 (5): 843-850 (2004). The reversibility analysis of apomorphine-induced motor activity (APO LOCO) was performed as described by Roux et al., Curr. Protocols in Pharmacol. Unit 5.17 (1999).

Summary of in vitro biological activity

Compound A competitively inhibits the binding of radiolabeled MLA to the rat brain hippocampus α7 NNR with equilibrium constant (Ki) values of ~ 1 nm, indicating that it has a very high affinity for the α7 NNR subtype. The stereoisomers of compound A have the following Ki values for rat α7 NNR: 2R, 3S (42 nM) [previously reported as 28 nM]; 2R, 3R (1 nM); 2S, 3S (25 μM) (see FIG. 1A). As illustrated in FIG. 1A2, compound A, enantiomer 2S, 3R, is active against the alpha7 subtype as opposed to its three other enantiomeric counterparts, which are represented as overlapping dots with weak activity. Compound A does not bind to α4β2 NNR with any significant affinity (Ki values> 2 μM).

Functional activity of (2S, 3R) -N- (2 - ((3-pyridinyl) methyl) -1-azabicyclo [2.2.2] oct-3-yl) benzofuran-2-carboxamide or its pharmaceutically acceptable salt (compound A) and its stereoisomers was investigated using a patch clamp (potential fixation) electrophysiological method for α7 NNR rat stably expressed in (mammalian) GH4C1 cells. In these experiments, compound A gave a strikingly different functional profile compared to other individual isomers and a racemic mixture of four isomers. As can be seen in FIGS. 1A and 1B, compound A is much stronger and more effective in eliciting a functional response (E _max = 93% relative to acetylcholine (ACh); EC ₅₀ = 14 nM) than any other isomer or mixture of four isomers. Indeed, compound A (isomer 2S, 3R) is the only isomer of N- (2 - ((3-pyridinyl) methyl) -1-azabicyclo [2.2.2] oct-3-yl) benzofuran-2-carboxamide, which can provide strong agonism in a concentration range of 1-50 nM, where a concentration of 10 nM refers to the in vivo activity described herein, as shown in Fig. 1B.

The functional activity of compound A was also evaluated electrophysiologically in Xenopus oocytes transiently expressing human α7 NNRs. In such a system, compound A has an EC ₅₀ value of 33 nM and E _{max of} 100% for the ACh response. A decrease in the subsequent control response for ACh was observed after administration of (2S, 3R) -N- (2 - ((3-pyridinyl) methyl) -1-azabicyclo [2.2.2] oct-3-yl) benzofuran-2-carboxamide in concentrations exceeding 100 nM (IC ₅₀ = 200 nM). In contrast to the previously described full α7 agonists (see Astles et al., Current Drug Targets CNS Neurological Disorders 1 (4): 337-348 (2002), incorporated herein by reference and relating to such a message), separation between EC values ₅₀ and IC ₅₀ show that concentrations that induce a half-maximal functional response of α7 lead to minimal rather than complete residual inhibition. There was no detectable activation when (2S, 3R) -N- (2 - ((3-pyridinyl) methyl) -1-azabicyclo [2.2.2] oct-3-yl) benzofuran-2-carboxamide was applied to oocytes expressing a subtype of human α4β2, and there was no significant decrease in subsequent control responses to ACh, which indicates that (2S, 3R) -N- (2 - ((3-pyridinyl) methyl) -1-azabicyclo [2.2.2 ] oct-3-yl) benzofuran-2-carboxamide is neither an agonist nor antagonist with respect to α4β2.

Compounds show little or no agonistic activity in functional models that use muscle-type receptors (subtype α1β1γδ in human TE671 / RD cloned cells) or ganglion-type receptors (subtype α3β4 subclone of the shooter cell of rat pheochromocytoma PC12 and cloned SHSY-5Y cells) , providing ≤10% (human muscle), ≤20% (rat ganglion) and ≤10% (human ganglion) nicotine response for these subtypes. These data show selectivity for CNS subtypes compared with other PNS (peripheral nervous system) subtypes.

Due to the close sequence homology and structural homology between the α7 and 5-hydroxytryptamine receptors (5HT ₃ ) and the cross-reactivity for these 2 receptors observed for other nicotinic receptors, the affinity of compound A for 5HT ₃ receptors was investigated. Compound A (10 μM) showed 59% inhibition of the mouse 5HT ₃ receptor and 25% inhibition of the human receptor in the radioactive binding assay. A study of functional activation on human 5HT ₃ receptors suggests minimal activation or lack of activation (i.e., a maximum response of 15% activation was observed at 100 μM).

Muscarinic receptors are another subject of consideration due to the interaction that has been observed for other nicotinic ligands. Compound A showed minimal or no interaction when tested in competitive binding inhibition assays for non-selective central and non-selective peripheral muscarinic receptors M1, M2.

The data show that compound A is selective for α7 NNR ligands. Compound A binds poorly to those subtypes of nicotinic receptors that are characteristic of the peripheral nervous system or to muscarinic or 5HT ₃ serotonergic receptors. Thus, compound A has a therapeutic potential for treating disorders of the central nervous system without causing side effects associated with interaction with the peripheral nervous system.

Summary of in vivo biological activity

Compound A, (2S, 3R) -N- (2 - ((3-pyridinyl) methyl) -1-azabicyclo [2.2.2] oct-3-yl) benzofuran-2-carboxamide or a pharmaceutically acceptable salt thereof exhibit significant effectiveness in two behavioral models of consciousness. Compound A showed high activity in the paradigm of object recognition in rats after i.p. (intraperitoneally, figure 3), and p.o. (oral administration, FIG. 4) administration and has also demonstrated activity over a wide dosage range after oral administration (FIG. 4). When administered intraperitoneally at the same low dose (0.3 and 1 mg / kg), compound A tends to reverse the deficiency caused by MK-801 in the PO problem (object recognition) (Fig. 5) and when administered orally at a dose of 0.3 mg / kg the cognitive effect of compound A lasts at least 18 hours (FIG. 6). In the paradigm of the labyrinth with an angular lever (RAM) (Fig. 7), which examines the working memory, connection A significantly increased the number of correct choices before an error. These results show the potential of (2S, 3R) -N- (2 - ((3-pyridinyl) methyl) -1-azabicyclo [2.2.2] oct-3-yl) benzofuran-2-carboxamide for the treatment of cognitive deficits and dysfunctions, associated with schizophrenia, including dysfunction of working memory.

In order for the compound to be used to treat cognitive dysfunction in schizophrenia, it should not reduce the effects of classic and atypical antipsychotics on the positive symptoms of schizophrenia. Thus, it is incontrovertible that, in addition to its cognitive enhancing properties, Compound A is also effective in reversing apomorphine-induced motor activity (APO LOCO) (FIG. 8) and prepulse inhibition (PPI) (FIG. 9) in positive models symptoms of schizophrenia. Thus, it can be expected that (2S, 3R) -N- (2 - ((3-pyridinyl) methyl) -1-azabicyclo [2.2.2] oct-3-yl) benzofuran-2-carboxamide will provide additional benefits in regarding positive as well as cognitive symptoms associated with schizophrenia.

The specific observed pharmacological responses may vary according to the particular active compound selected, whether it contains pharmaceutical carriers, as well as the type of preparation and the route of administration used, and such expected deviations or differences in results are considered in accordance with the practical implementation of the present invention.

Although specific embodiments of the present invention are illustrated and described in detail here, the invention is not limited to them. The above detailed descriptions are provided to illustrate the present invention and should not be construed as constituting any limitation of the invention. Modifications will be apparent to those skilled in the art, and it is intended that all modifications that do not depart from the spirit of the invention are included within the scope of the appended claims.

Claims (5)

1. The acid salt and (2S, 3R) -N- (2 - ((3-pyridinyl) methyl) -1-azabicyclo [2.2.2] oct-3-yl) benzofuran-2-carboxamide, essentially free of (2S, 3S) -N- (2 - ((3-pyridinyl) methyl) -1-azabicyclo [2.2.2] oct-3-yl) benzofuran-2-carboxamide, (2R, 3S) -N- (2 - ((3-pyridinyl) methyl) -1-azabicyclo [2.2.2] oct-3-yl) benzofuran-2-carboxamide and (2R, 3R) -N- (2 - ((3-pyridinyl) methyl) - 1-azabicyclo [2.2.2] oct-3-yl) benzofuran-2-carboxamide, where the enantiomeric or diastereomeric excess of (2S, 3R) -N- (2 - ((3-pyridinyl) methyl) -1-azabicyclo [2.2 .2] oct-3-yl) benzofuran-2-carboxamide selected from 90% or more; 95% or more; 98% or more; 99% or more; or 99.5% or more, and wherein the acid is selected from hydrochloric acid, sulfuric acid, phosphoric acid, maleic acid, toluenesulfonic acid, galactic (mucus) acid, D-mandelic acid, D-tartaric acid, methanesulfonic acid, R- and S-10-camphorsulfonic acid, ketoglutaric acid or hippuric acid. 2. The compound according to claim 1, where the stoichiometric ratio of (2S, 3R) -N- (2 - ((3-pyridinyl) methyl) -1-azabicyclo [2.2.2] oct-3-yl) benzofuran-2-carboxamide to acid is 2: 1, 1: 1 or 1: 2. 3. The compound according to claim 2, where the stoichiometric ratio is 1: 1. 4. (2S, 3R) -N- (2 - ((3-pyridinyl) methyl) -1-azabicyclo [2.2.2] oct-3-yl) benzofuran-2-carboxamide hydrochloride substantially free of hydrochloride ( 2S, 3S) -N- (2 - ((3-pyridinyl) methyl) -1-azabicyclo [2.2.2] oct-3-yl) benzofuran-2-carboxamide, hydrochloride (2R, 3S) -N- (2 - ((3-pyridinyl) methyl) -1-azabicyclo [2.2.2] oct-3-yl) benzofuran-2-carboxamide and (2R, 3R) -N- (2 - ((3-pyridinyl) methyl) hydrochloride -1-azabicyclo [2.2.2] oct-3-yl) benzofuran-2-carboxamide, where the enantiomeric or diastereomeric excess of (2S, 3R) -N- (2 - ((3-pyridinyl) methyl) -1-azabicyclo [ 2.2.2] oct-3-yl) benzofuran-2-carboxamide selected from 90% or more; 95% or more; 98% or more; 99% or more; or 99.5% or more. 5. (2S, 3R) -N- (2 - ((3-pyridinyl) methyl) -1-azabicyclo [2.2.2] oct-3-yl) benzofuran-2-carboxamide monohydrochloride or a hydrate or solvate thereof, essentially free of (2S, 3S) -N- (2 - ((3-pyridinyl) methyl) -1-azabicyclo [2.2.2] oct-3-yl) benzofuran-2-carboxamide monohydrochloride, monohydrochloride (2R, 3S) -N- (2 - ((3-pyridinyl) methyl) -1-azabicyclo [2.2.2] oct-3-yl) benzofuran-2-carboxamide and monohydrochloride (2R, 3R) -N- (2 - ((3 -pyridinyl) methyl) -1-azabicyclo [2.2.2] oct-3-yl) benzofuran-2-carboxamide, where enantiomeric or diastereomeric excess (2S, 3R) -N- (2 - ((3-pyridinyl) methyl) -1-azabicyclo [2.2.2] oct-3-yl) benzofuran-2-car oxamide selected from 90% or more; 95% or more; 98% or more; 99% or more; or 99.5% or more.

Images