Pharmaceutical Compounds

Abstract

The present invention relates to compounds which are agonists of the muscarinic M1 and M4 receptors and are useful in the treatment of diseases mediated by the muscarinic M1 and / or M4 receptors. Also provided are pharmaceutical compositions containing said compounds and therapeutic uses of said compounds. TIFF2024504252000051.tif3653(1) and salts thereof.

Description

[Technical Field]

[0001] The present invention relates to a class of novel heterocyclic compounds, their salts, pharmaceutical compositions containing them, and their use in treating the human body. In particular, the present invention relates to a class of compounds that are agonists of the muscarinic M1 and / or M4 receptors and are therefore useful in the treatment of Alzheimer's disease, schizophrenia, cognitive impairment, and other diseases mediated by the muscarinic M1 / M4 receptors, including, but not limited to, the treatment or alleviation of pain. [Background technology]

[0002] Muscarinic acetylcholine receptors (mAChRs) are members of the G protein-coupled receptor superfamily that mediate the actions of the neurotransmitter acetylcholine in both the central and peripheral nervous systems. Five mAChR subtypes, M1-M5, have been cloned. M1 mAChRs are expressed postsynaptically primarily in the cerebral cortex, hippocampus, striatum, and thalamus; M2 mAChRs are present primarily in the brainstem and thalamus, but also in the cerebral cortex, hippocampus, and striatum; and M2 mAChRs are present at cholinergic synaptic terminals (Langmead et al., 2008 Br J Pharmacol). However, M2 mAChRs are also expressed peripherally in cardiac tissue (where they mediate vagal innervation of the heart) and in smooth muscle and exocrine glands. M3mAChR is expressed at relatively low levels in the CNS, but is widely expressed in smooth muscle and glandular tissues such as sweat and salivary glands (Langmead et al., 2008 Br J Pharmacology). Muscarinic receptors in the central nervous system, particularly M1 mAChRs, play an important role in mediating higher-level cognitive processing. Diseases associated with cognitive impairment, such as Alzheimer's disease, are associated with the loss of cholinergic neurons in the basal forebrain (Whitehouse et al., 1982 Science). Schizophrenia, also characterized by cognitive dysfunction, also shows reduced mAChR density in the prefrontal cortex, hippocampus, and caudate-putamen of schizophrenic subjects (Dean et al., 2002 Mol Psychiatry). Furthermore, in animal models, blockade or damage to central cholinergic pathways has been shown to cause severe cognitive impairment, and nonselective mAChR antagonists have been shown to induce psychotomimetic effects in psychiatric patients. Cholinergic replacement therapy is primarily based on the use of acetylcholinesterase inhibitors to prevent the breakdown of endogenous acetylcholine. These compounds have shown clinical efficacy in treating symptomatic cognitive decline, but cause dose-limiting side effects (e.g., gastrointestinal motility disorders, bradycardia, nausea, and vomiting) due to stimulation of peripheral M2 and M3 mAChRs (http: / / www.drugs.com / pro / donepezil.html; http: / / www.drugs.com / pro / rivastigmine.html).

[0003] Furthermore, there has been a growing search for direct M1 mAChR agonists that target cognitive enhancement. These efforts have resulted in the identification of various agonists, including compounds such as xanomeline, AF267B, sabcomeline, miramelin, and cevimeline. Many of these compounds have been shown to be highly effective in preclinical models of cognition in both rodents and / or non-human primates. Miramelin has demonstrated efficacy against scopolamine-induced working and spatial memory impairments in rodents, sabcomeline has demonstrated efficacy in a visual object discrimination task in marmosets, and xanomeline has reversed mAChR antagonist-induced impairments in cognitive performance in a passive avoidance paradigm. Alzheimer's disease (AD) is the most common neurodegenerative disorder affecting older adults (26.6 million people worldwide in 2006), resulting in severe memory loss and cognitive impairment. The pathogenesis of this disease is complex but characterized by two distinct brain sequelae: the aggregation of amyloid plaques, primarily composed of amyloid-β peptide (Aβ), and neurofibrillary tangles, formed by hyperphosphorylation of tau protein. Aβ accumulation is considered a central feature of AD progression; therefore, many putative therapies for the treatment of AD currently target the inhibition of Aβ production. Aβ is derived from the proteolytic cleavage of membrane-bound amyloid precursor protein (APP). APP is processed via two pathways: the nonamyloidogenic pathway and the amyloidogenic pathway. While cleavage of APP by γ-secretase is common to both pathways, in the former, APP is cleaved by α-secretase to produce soluble APPα. The cleavage site is within the Aβ sequence, preventing the formation of Aβ. However, in the amyloidogenic pathway, APP is also cleaved by β-secretase to produce soluble APPβ and Aβ. In vitro studies have shown that mAChR agonists can promote the processing of APP to the soluble, nonamyloidogenic pathway. In vivo studies have shown that the mAChR agonist AF267B altered disease-like pathology in 3×TgAD transgenic mice, a model of various components of Alzheimer's disease (Caccamo et al., 2006 Neuron). Finally, the mAChR agonist cevimeline has been shown to slightly but significantly reduce cerebrospinal fluid levels of Aβ in patients with Alzheimer's disease, thus demonstrating potential disease-modifying efficacy (Nitsch et al., 2000 Neurol).

[0004] Furthermore, preclinical studies suggest that mAChR agonists exhibit atypical antipsychotic-like profiles in various preclinical paradigms. The mAChR agonist xanomeline reverses many dopamine-driven behaviors (e.g., amphetamine-induced locomotion in rats, apomorphine-induced climbing in mice, dopamine agonist-driven rotation in unilateral 6-OH-DA-lesioned rats, and amphetamine-induced locomotor anxiety in monkeys) without EPS tendency. Xanomeline has also been shown to inhibit A10, but not A9, dopamine cell firing and conditioned avoidance, and induces c-fos expression in the prefrontal cortex and nucleus accumbens, but not in the striatum, of rats. All of these data suggest an atypical antipsychotic-like profile (Mirza et al., 1999 CNS Drug Rev). Muscarinic receptors are also implicated in the neurobiology of addiction. The reinforcing effects of cocaine and other addictive substances are mediated by the mesolimbic dopamine system, where behavioral and neurochemical studies have shown that cholinergic muscarinic receptor subtypes play an important role in regulating dopaminergic neurotransmission. For example, M(4)(- / -) mice exhibited significantly enhanced reward-driven behavior as a result of exposure to cocaine (Schmidt et al. Psychopharmacology (2011) Aug;216(3):367-78). Furthermore, xanomeline has been demonstrated to block the effects of cocaine in these models.

[0005] Muscarinic receptors are also involved in the control of movement and may represent novel therapeutic agents for movement disorders such as Parkinson's disease, ADHD, Huntington's disease, Tourette's syndrome, and other syndromes associated with dopaminergic dysfunction as the underlying pathogenic factor driving the disease. Xanomeline, sabcomeline, miramelin, and cevimeline are all in various stages of clinical development for the treatment of Alzheimer's disease and / or schizophrenia. Phase II clinical studies of xanomeline demonstrated its efficacy in various cognitive symptom domains, including behavioral disturbances and hallucinations associated with Alzheimer's disease (Bodick et al., 1997 Arch Neurol). This compound was also evaluated in a small Phase II study of schizophrenic patients, where it significantly reduced positive and negative symptoms compared with placebo controls (Shekhar et al., 2008 Am J Psych). However, in all clinical studies, xanomeline and other related mAChR agonists demonstrated unacceptable safety margins with respect to cholinergic side effects, such as nausea, gastrointestinal pain, diarrhea, diaphoresis (excessive sweating), hypersalivation (excessive salivation), syncope, and bradycardia.

[0006] Muscarinic receptors are involved in central and peripheral pain. Pain can be divided into three distinct types: acute, inflammatory, and neuropathic. Acute pain plays an important protective role, protecting the body from stimuli that may cause tissue damage, but pain management is also necessary after surgery. Inflammatory pain can arise from many causes, including tissue injury, autoimmune responses, and pathogen invasion, and is triggered by the action of inflammatory mediators such as neuropeptides and prostaglandins, which cause nerve inflammation and pain. Neuropathic pain involves abnormal pain sensations in response to innocuous stimuli. Neuropathic pain is associated with many different diseases / injuries, including spinal cord injury, multiple sclerosis, diabetes (diabetic neuropathy), and viral infections (e.g., HIV or herpes). It is also commonly seen in cancer, both as a result of disease or as a side effect of chemotherapy. Activation of muscarinic receptors has been shown to have analgesic effects for many pain conditions through activation of receptors in higher pain centers in the brain and spinal cord. Elevating endogenous levels of acetylcholinesterase inhibitors and direct activation of muscarinic receptors by agonists or allosteric modulators have been shown to have analgesic activity. In contrast, blocking muscarinic receptors with antagonists or by using knockout mice increases pain sensitivity. Evidence for the role of M1 receptors in pain is reviewed in D.F. Fiorino and M. Garcia-Guzman, 2012. More recently, a small number of compounds have been identified that exhibit improved selectivity for the M1 mAChR subtype relative to peripherally expressed mAChR subtypes (Bridges et al., 2008 Bioorg Med Chem Lett; Johnson et al., 2010 Bioorg Med Chem Lett; Budzik et al., 2010 ACS Med Chem Lett). Despite the increased level of selectivity for the M3 mAChR subtype, some of these compounds retain significant agonist activity at both this subtype and the M2 mAChR subtype. Herein, we describe a series of compounds that unexpectedly exhibit high levels of selectivity for the M1 and / or M4 mAChR relative to the M2 and M3 receptor subtypes. Summary of the Invention

[0007] The present invention provides compounds that have activity as muscarinic M1 and / or M4 receptor agonists. More specifically, the present invention provides compounds that exhibit selectivity for the M1 and / or M4 receptor over the M2 and M3 receptor subtypes. Thus, the present invention provides a compound of formula (1): [ka] (1) or a salt thereof. The present invention relates to a compound of formula (1a): [ka] (1a) The present invention further provides a compound of the formula (wherein X represents a salt). The present invention relates to a compound of formula (1b): [ka] (1b) The present invention further provides a compound of the formula:

[0008] The present invention relates to a compound of formula (2): [ka] (2) or a salt thereof. The present invention relates to a compound of formula (2a): [ka] (2a) The present invention further provides a compound of the formula (wherein X represents a salt).

[0009] The present invention relates to a compound of formula (2b): [ka] (2b) The present invention further provides a compound of the formula: The present invention relates to a compound of formula (2c): [ka] (2c) The present invention further provides a compound of the formula:

[0010] The compound of formula (1) or formula (2) may be a pharmaceutically acceptable salt. The compounds of formula (1) or formula (2) may be acid addition salts. The compound of formula (1) or formula (2) may be a hydrochloride salt. The compound of formula (1) or formula (2) may be a hydrochloride salt. The compound of formula (1) or formula (2) may be the monohydrochloride monohydrate. In the compounds of formula (1a) or formula (2a), X can be a pharmaceutically acceptable salt. In the compounds of formula (1a) or formula (2a), X may be an acid addition salt. In the compounds of formula (1a) or formula (2a), X may be a hydrochloride salt. In the compounds of formula (1a) or formula (2a), X may be a monohydrochloride salt. The compound of formula (1) or formula (2) may be a hydrochloride salt. X can be the hydrochloride salt. X can be the monohydrochloride salt. X can be the monohydrochloride monohydrate. The compound can be N-tert-butyl-1-{8-[3-(trifluoromethyl)-1,2,4-oxadiazol-5-yl]-8-azabicyclo[3.2.1]octan-3-yl}piperidine-4-carboxamide. The compound can be N-tert-butyl-1-{(1R,3r,5S)-8-[3-(trifluoromethyl)-1,2,4-oxadiazol-5-yl]-8-azabicyclo[3.2.1]octan-3-yl}piperidine-4-carboxamide. The compound can be a salt of N-tert-butyl-1-{8-[3-(trifluoromethyl)-1,2,4-oxadiazol-5-yl]-8-azabicyclo[3.2.1]octan-3-yl}piperidine-4-carboxamide. The compound can be a salt of N-tert-butyl-1-{(1R,3r,5S)-8-[3-(trifluoromethyl)-1,2,4-oxadiazol-5-yl]-8-azabicyclo[3.2.1]octan-3-yl}piperidine-4-carboxamide.

[0011] The compound can be a pharmaceutically acceptable salt of N-tert-butyl-1-{8-[3-(trifluoromethyl)-1,2,4-oxadiazol-5-yl]-8-azabicyclo[3.2.1]octan-3-yl}piperidine-4-carboxamide. The compound can be a pharmaceutically acceptable salt of N-tert-butyl-1-{(1R,3r,5S)-8-[3-(trifluoromethyl)-1,2,4-oxadiazol-5-yl]-8-azabicyclo[3.2.1]octan-3-yl}piperidine-4-carboxamide. The compound can be N-tert-butyl-1-{8-[3-(trifluoromethyl)-1,2,4-oxadiazol-5-yl]-8-azabicyclo[3.2.1]octan-3-yl}piperidine-4-carboxamide hydrochloride. The compound can be N-tert-butyl-1-{(1R,3r,5S)-8-[3-(trifluoromethyl)-1,2,4-oxadiazol-5-yl]-8-azabicyclo[3.2.1]octan-3-yl}piperidine-4-carboxamide hydrochloride. The compound can be N-tert-butyl-1-{8-[3-(trifluoromethyl)-1,2,4-oxadiazol-5-yl]-8-azabicyclo[3.2.1]octan-3-yl}piperidine-4-carboxamide monohydrochloride. The compound can be N-tert-butyl-1-{(1R,3r,5S)-8-[3-(trifluoromethyl)-1,2,4-oxadiazol-5-yl]-8-azabicyclo[3.2.1]octan-3-yl}piperidine-4-carboxamide monohydrochloride. The compound can be N-tert-butyl-1-{8-[3-(trifluoromethyl)-1,2,4-oxadiazol-5-yl]-8-azabicyclo[3.2.1]octan-3-yl}piperidine-4-carboxamide monohydrochloride monohydrate. The compound can be N-tert-butyl-1-{(1R,3r,5S)-8-[3-(trifluoromethyl)-1,2,4-oxadiazol-5-yl]-8-azabicyclo[3.2.1]octan-3-yl}piperidine-4-carboxamide monohydrochloride monohydrate. DETAILED DESCRIPTION OF THE INVENTION

[0012] definition As used herein, the following definitions apply unless otherwise indicated. The term "treatment," with respect to the use of a compound of Formula (1), Formula (1a), Formula (1b), Formula (2), Formula (2a), Formula (2b), or Formula (2c), is used to describe any form of intervention in which a compound is administered to a subject suffering from, at risk of suffering from, or potentially at risk of suffering from the disease or disorder in question. Thus, the term "treatment" encompasses both prophylactic treatment and treatment in which there are measurable or detectable symptoms of the disease or disorder. As used herein, the term "therapeutically effective amount" (e.g., with respect to a method of treating a disease or condition) refers to an amount of a compound that is effective to produce a desired therapeutic effect. For example, if the condition is pain, a therapeutically effective amount is an amount sufficient to provide a desired level of pain relief. The desired level of pain relief can be, for example, complete elimination of pain or a reduction in the severity of pain.

[0013] salt The compounds described herein can exist in the form of salts, such as acid addition salts, and in some cases salts of organic and inorganic bases, such as carboxylates, sulfonates, and phosphates. All such salts are within the scope of the present invention, and references to compounds of formula (1) and formula (2) include salt forms of the compounds defined herein. The salts are typically acid addition salts. The salts of the present invention can be synthesized from a parent compound that contains a basic or acidic moiety by conventional chemical methods, such as those described in Pharmaceutical Salts: Properties, Selection, and Use, P. Heinrich Stahl (Editor), Camille G. Wermuth (Editor), ISBN: 3-90639-026-8, Hardcover, 388 pages, August 2002. Generally, such salts can be prepared by reacting the free acid or free base form of these compounds with the appropriate base or acid in water or an organic solvent, or in a mixture of both, generally using a non-aqueous medium such as ether, ethyl acetate, ethanol, isopropanol, or acetonitrile.

[0014] Acid addition salts can be formed with a wide variety of acids, both organic and inorganic. Examples of acid addition salts within the scope of the present invention include acetic acid, 2,2-dichloroacetic acid, adipic acid, alginic acid, ascorbic acid (e.g., L-ascorbic acid), L-aspartic acid, benzenesulfonic acid, benzoic acid, 4-acetamidobenzoic acid, butanoic acid, (+) camphoric acid, camphorsulfonic acid, (+)-(1S)-camphor-10-sulfonic acid, capric acid, caproic acid, caprylic acid, and cinnamic acid. , citric acid, cyclamic acid, dodecyl sulfate, ethane-1,2-disulfonic acid, ethanesulfonic acid, 2-hydroxyethanesulfonic acid, formic acid, fumaric acid, galactaric acid, gentisic acid, glucoheptonic acid, D-gluconic acid, glucuronic acid (e.g., D-glucuronic acid), glutamic acid (e.g., L-glutamic acid), α-oxoglutaric acid, glycolic acid, hippuric acid, hydrohalic acids (e.g., bromide, HCl), acid, hydrochloric acid, hydroiodic acid), isethionic acid, lactic acid (e.g., (+)-L-lactic acid, (±)-DL-lactic acid), lactobionic acid, maleic acid, malic acid, (-)-L-malic acid, malonic acid, (±)-DL-mandelic acid, methanesulfonic acid, naphthalene-2-sulfonic acid, naphthalene-1,5-disulfonic acid, 1-hydroxy-2-naphthoic acid, nicotinic acid, nitric acid, oleic acid, orotic acid, oxalic acid, palmitic acid, Examples of suitable salts include mono- or di-salts formed with acids selected from the group consisting of mitic acid, pamoic acid, phosphoric acid, propionic acid, pyruvic acid, L-pyroglutamic acid, salicylic acid, 4-amino-salicylic acid, sebacic acid, stearic acid, succinic acid, sulfuric acid, tannic acid, (+)-L-tartaric acid, thiocyanic acid, p-toluenesulfonic acid, undecylenic acid, and valeric acid, as well as mono- or di-salts formed with acylated amino acids and cation exchange resins.

[0015] Amine functional groups in the compounds described herein may be reacted with, for example, alkylating agents to form quaternary ammonium salts, according to methods well known to those skilled in the art. Such quaternary ammonium compounds are within the scope of the present invention. The compounds of the present invention may exist as mono- or di-salts depending on the pKa of the acid forming the salt. The salt forms of the compounds of the present invention are typically pharmaceutically acceptable salts, and examples of pharmaceutically acceptable salts are discussed in Berge et al., 1977, "Pharmaceutically Acceptable Salts," J. Pharm. Sci., Vol. 66, pp. 1-19. However, pharmaceutically unacceptable salts can also be prepared as intermediate forms and then converted into pharmaceutically acceptable salts. Such pharmaceutically unacceptable salt forms may be useful, for example, in the purification or separation of the compounds of the present invention, and also form part of the present invention.

[0016] stereoisomer References to compounds of formula (1), (1a) and (1b) include all possible stereoisomeric forms thereof (e.g., enantiomers, epimers and diastereomers; endo-exo isomers, etc.), either as individual isomers or mixtures (e.g., racemic mixtures) or two or more isomers, unless the context requires otherwise. Thus, the present invention provides compounds of formula (1) which contain a chiral center. Isomers can be characterized in terms of their absolute stereochemistry using the "R and S" nomenclature developed by Cahn, Ingold, and Prelog. Advanced Organic Chemistry by Jerry March, 4 thEdition, John Wiley & Sons, New York, 1992, pages 109-114, and Cahn, Ingold & Prelog, Angew. Chem. Int. Ed. Engl., 1966, 5, 385-415. Stereoisomers can be separated by a number of techniques, including chiral chromatography (chromatography on a chiral support), and such techniques are well known to those skilled in the art. As an alternative to chiral chromatography, stereoisomers can be separated by forming diastereomeric salts with chiral acids such as (+)-tartaric acid, (-)-pyroglutamic acid, (-)-di-toluoyl-L-tartaric acid, (+)-mandelic acid, (-)-malic acid, and (-)-camphorsulfonic acid, followed by separation of the diastereomers by selective crystallization, followed by dissociation of the salts to give the individual enantiomers of the free base.

[0017] When a compound of the invention exists in two or more stereoisomeric forms, one diastereomer of a pair may be superior to the other diastereomer, for example, in terms of biological activity, and therefore, in certain situations, it may be desirable to use only one of the diastereomers as a therapeutic agent. Accordingly, the present invention provides a composition comprising a compound having one or more chiral centers, wherein at least 55% (e.g., at least 60%, 65%, 70%, 75%, 80%, 85%, 90%, or 95%) of the compound is present as a single isomer (e.g., a diastereomer). In one typical embodiment, 99% or more (eg, substantially all) of the total amount of the compound (or compound for use) is present as a single isomer. For example, in one embodiment, the compound exists as a single diastereomer, and the compound possesses a plane of symmetry.

[0018] Isotopes The compounds of the present invention may contain one or more isotopic substitutions, and a reference to a particular element includes within its scope all isotopes of that element. For example, a reference to hydrogen includes within its scope 1 H, 2 H(D), and 3 Similarly, when carbon and oxygen are mentioned, the respective ranges include H(T). 12 C. 13 C and 14 C and 16 O and 18 Contains O. Similarly, when a particular functional group is referred to, isotopic variations are also included within its scope unless the context indicates otherwise. For example, reference to an alkyl group such as a tert-butyl group also encompasses variations in which one or more of the hydrogen atoms in the group are in the form of deuterium or tritium (tritium) isotopes, such as a tert-butyl group in which all nine hydrogen atoms are in the form of deuterium isotopes (perdeutero-tert-butyl group). The isotope may be radioactive or non-radioactive. The compound may not contain a radioisotope. Such compounds are preferred for therapeutic use. However, the compound may contain one or more radioisotopes. Compounds containing such radioisotopes may be useful in diagnostic settings.

[0019] solvate The compounds of the present invention can form solvates. Preferred solvates are those formed by incorporating molecules of a non-toxic, pharmaceutically acceptable solvent (hereinafter referred to as a solvating solvent) into the solid structure (e.g., crystalline structure) of a compound of the present invention. Examples of such solvents include water, alcohols (e.g., ethanol, isopropanol, and butanol), and dimethyl sulfoxide. Solvates can be prepared by recrystallizing a compound of the present invention from a solvent or a mixture of solvents containing a solvating solvent. In any case, whether a solvate has been formed can be determined by analyzing crystals of the compound using well-known and standard techniques, such as thermogravimetric analysis (TGE), differential scanning calorimetry (DSC), and X-ray crystallography. Solvates may be stoichiometric or non-stoichiometric. Particularly preferred solvates are hydrates, and examples of hydrates include hemihydrates, monohydrates, and dihydrates. Therefore, the present invention provides the compound in the form of a solvate, a compound wherein the solvate is a hydrate; The solvate is a monohydrate. For a more detailed discussion of solvates and the methods used to prepare and characterize them, see Bryn et al., Solid-State Chemistry of Drugs, Second Edition (ISBN 0-967-06710-3), published by SSCI, Inc. of West Lafayette, IN, USA, 1999. Alternatively, the compounds of the invention may be anhydrous, rather than existing as hydrates. Accordingly, the invention provides compounds of the invention in anhydrous form (e.g., anhydrous crystalline form).

[0020] Crystalline and amorphous forms A compound may exist in a crystalline or non-crystalline (e.g., amorphous) state. Whether a compound exists in a crystalline state can be readily determined by standard techniques, such as X-ray powder diffraction (XRPD). Crystals and their crystalline structures can be characterized using a number of techniques, including single crystal X-ray crystallography, X-ray powder diffraction (XRPD), differential scanning calorimetry (DSC), and infrared spectroscopy (e.g., Fourier transform infrared spectroscopy (FTIR)). The behavior of crystals under varying humidity conditions can also be analyzed by XRPD through gravimetric vapor sorption studies. The crystalline structure of a compound can be determined by X-ray crystallography, which can be carried out according to conventional methods, such as those described herein and in Fundamentals of Crystallography, C. Giacovazzo, H.L. Monaco, D. Viterbo, F. Scordari, G. Gilli, G. Zanotti, and M. Catti (International Union of Crystallography / Oxford University Press, 1992 ISBN 0-19-855578-4 (p / b), 0-19-85579-2 (h / b)). This technique involves the analysis and interpretation of X-ray diffraction patterns of single crystals. In amorphous solids, the three-dimensional structure typically present in crystalline forms does not exist, and the relative positions of molecules in amorphous forms are essentially random. See, for example, Hancock et al. J. Pharm. Sci. (1997), 86, 1). Therefore, the present invention provides the compound in crystalline form, (a) a compound that is 50% to 100% crystalline, more specifically, at least 50% crystalline, or at least 60% crystalline, or at least 70% crystalline, or at least 80% crystalline, or at least 90% crystalline, or at least 95% crystalline, or at least 98% crystalline, or at least 99% crystalline, or at least 99.5% crystalline, or at least 99.9% crystalline, e.g., 100% crystalline; The compound is provided in an amorphous form.

[0021] Complexes and clathrates The compounds of the present invention also include complexes (eg, inclusion complexes or clathrates with compounds such as cyclodextrins, or complexes with metals). Thus, the present invention provides compounds in the form of complexes or clathrates. Biological Activities and Therapeutic Uses The compounds of the present invention have activity as muscarinic M1 and M4 receptor agonists. The muscarinic activity of the compounds can be determined using the phosphorylation-ERK1 / 2 assay described in Example A below. A significant advantage of the compounds of the present invention is that they are highly selective for the M1 and / or M4 receptors compared to the M2 and M3 receptor subtypes. The compounds of the present invention are not agonists of the M2 and M3 receptor subtypes. For example, the compounds of the present invention typically have a pEC50 of at least 6 (preferably at least 6.5) for the M1 receptor in the functional assay described in Example A. 50 value and E of more than 80 (preferably more than 90) max values, while having a pEC value of less than 5 when tested against M2 and M3 subtypes in the functional assay of Example A. 50 value and less than 20% E max It may have a value.

[0022] With respect to the compounds of the present invention, the present invention further provides: compounds for use in medicine, Compounds for use as muscarinic M1 and / or M4 receptor agonists, a pEC50 of greater than 6.9 for the M1 receptor in the assay of Example A herein or a substantially similar assay 50 and at least 80 E max a compound which is a muscarinic M1 receptor agonist having the formula: pEC > 7.0 50 a compound which is a muscarinic M1 receptor agonist having the formula: At least 90 E for M1 receptors max a compound having a pEC50 of greater than 6.0 for the M4 receptor in the assay of Example A herein or a substantially similar assay 50 a compound which is a muscarinic M1 and M4 receptor agonist having the formula: compounds that are selective for muscarinic M1 and M4 receptors relative to M2 and M3 receptors; pEC50 of less than 5 for muscarinic M2 and M3 receptor subtypes 50 and E less than 30 max a compound having A compound for use in the treatment of a disease or condition mediated by muscarinic M1 and / or M4 receptors. Due to their muscarinic M1 and M4 receptor agonist activity, the compounds of the present invention can be used in the treatment of Alzheimer's disease, dementia with Lewy bodies, schizophrenia, and other psychiatric disorders, cognitive disorders, and other diseases mediated by muscarinic M1 and / or M4 receptors, and can also be used in the treatment of various types of pain.

[0023] Thus, with respect to the compounds of the present invention, the present invention further provides: a compound for use in the treatment of a cognitive or psychiatric disorder; 1. A compound for use in treating a cognitive or psychiatric disorder, wherein the cognitive or psychiatric disorder comprises, results from, or is associated with a condition selected from the following: cognitive impairment, mild cognitive impairment (MCI) (including mild cognitive impairment due to Alzheimer's disease and / or prodromal Alzheimer's disease, such as amnestic MCI and non-amnestic MCI), frontotemporal dementia, vascular dementia, dementia with Lewy bodies, presenile dementia, senile dementia, Friedreich's ataxia, Down's syndrome, Huntington's chorea, hyperkinesia, mania, Tourette's syndrome, Alzheimer's disease (prodromal Alzheimer's disease), and conditions listed in the U.S. Food and Drug Administration's "Early Alzheimer's disease: Developing Drugs for Treatment" (fda.gov / downloads / Drugs / GuidanceComplianceRegulatoryInformation / Guidances / UCM596728).pdf available), progressive supranuclear palsy, cognitive impairment (such as attention, orientation, learning disabilities, memory (i.e., memory impairment, amnesia, amnesic disorder, transient global amnesia and age-related memory impairment), and language function); cognitive impairment as a result of stroke, Huntington's disease, Pick's disease, AIDS-related dementia or other dementia conditions, e.g., multi-infarct dementia, alcoholic dementia, hypothyroidism-related dementia, and dementias associated with other degenerative disorders such as cerebellar atrophy and amyotrophic lateral sclerosis; other acute or subacute conditions that may cause cognitive decline, such as delirium or depression (pseudo-dementia states), trauma, head injury, age-related cognitive decline, stroke, neurodegeneration, drug-induced conditions, cognitive impairment due to neurotoxic drugs, age-related cognitive impairment, autism-related cognitive impairment, Down's syndrome, psychosis-related cognitive impairment, and cognitive impairment following electroconvulsive therapy; cognitive impairment due to substance abuse or withdrawal (drugs such as nicotine, cannabis, amphetamines, and cocaine); attention deficit hyperactivity disorder (ADHD) and movement disorders (e.g., Parkinson's disease, neuroleptic-induced parkinsonism, and tardive dyskinesia); schizophrenia, schizophreniform disorders, psychotic depression, mania, acute mania, paranoid, hallucinatory, and delusional disorders; personality disorders, obsessive-compulsive disorder, schizotypal disorder, delusional disorder, psychosis due to malignancy, or metabolic disorders, or endocrine disorders or narcolepsy; psychosis due to substance abuse or withdrawal, bipolar disorder, and schizoaffective disorder.

[0024] A compound for use in treating Alzheimer's disease, A compound for use in the treatment of dementia with Lewy bodies, a compound for use in the treatment of schizophrenia, a method of treating a cognitive disorder in a subject (e.g., a mammalian patient, such as a human (e.g., a human in need of treatment)), comprising administering a therapeutically effective amount of a compound of the invention; a method for treating a cognitive impairment in a subject (e.g., a mammalian patient such as a human (e.g., a human in need of treatment)), comprising administering a therapeutically effective amount of a compound of the invention, wherein the cognitive impairment comprises, results from, or is associated with a condition as defined above; a method for treating cognitive impairment in a subject (e.g., a mammalian patient, such as a human (e.g., a human in need of treatment)), comprising administering a therapeutically effective amount of a compound of the invention, wherein the cognitive impairment results from or is associated with Alzheimer's disease; a method for treating a cognitive disorder in a subject (e.g., a mammalian patient such as a human (e.g., a human in need of treatment)), comprising administering a therapeutically effective amount of a compound of the invention, wherein the cognitive disorder is dementia with Lewy bodies; a method for treating a cognitive disorder in a subject (e.g., a mammalian patient, such as a human (e.g., a human in need of treatment)), comprising administering a therapeutically effective amount of a compound of the invention, wherein the cognitive disorder is schizophrenia;

[0025] Use of a compound of the present invention for the manufacture of a medicament for the treatment of cognitive disorders; Use of a compound of the invention for the manufacture of a medicament for the treatment of a cognitive disorder, wherein the cognitive disorder comprises, results from or is associated with a condition as defined above; Use of a compound of the present invention for the manufacture of a medicament for the treatment of cognitive impairment, wherein the cognitive impairment includes, results from, or is associated with Alzheimer's disease; Use of a compound of the present invention for the manufacture of a medicament for the treatment of cognitive disorders, wherein the cognitive disorders include, result from, or are associated with dementia with Lewy bodies; Use of a compound of the present invention for the manufacture of a medicament for the treatment of a cognitive disorder, wherein the cognitive disorder comprises, results from, or is associated with schizophrenia; Compounds for the treatment or reduction of severity of acute, chronic, neuropathic or inflammatory pain, arthritis, migraine, cluster headache, trigeminal neuralgia, herpetic neuralgia, general neuralgia, visceral pain, osteoarthritic pain, post-herpetic neuralgia, diabetic neuropathy, radicular pain, sciatica, back pain, headache or neck pain, severe or intractable pain, nociceptive pain, breakthrough pain, post-operative pain, or cancer pain,

[0026] 1. A method for treating or reducing the severity of acute, chronic, neuropathic, or inflammatory pain, arthritis, migraine, cluster headache, trigeminal neuralgia, herpetic neuralgia, generalized neuralgia, visceral pain, osteoarthritic pain, postherpetic neuralgia, diabetic neuropathy, radicular pain, sciatica, back pain, headache or neck pain, severe or intractable pain, nociceptive pain, breakthrough pain, post-operative pain, or cancer pain, which method comprises administering a therapeutically effective amount of a compound of the present invention; compounds for the treatment of peripheral disorders such as reducing intraocular pressure in glaucoma, and for the treatment of dry eye and dry mouth such as Sjogren's syndrome; a method for treating peripheral disorders such as reducing intraocular pressure in glaucoma, and dry eye and dry mouth such as Sjogren's syndrome, which comprises administering a therapeutically effective amount of a compound of the present invention; use of a compound of the invention for the manufacture of a medicament for the treatment or lessening of the severity of acute, chronic, neuropathic or inflammatory pain, arthritis, migraine, cluster headache, trigeminal neuralgia, herpetic neuralgia, generalized neuralgia, visceral pain, osteoarthritic pain, post-herpetic neuralgia, diabetic neuropathy, radicular pain, sciatica, back pain, headache or neck pain, severe or intractable pain, nociceptive pain, breakthrough pain, post-operative pain or cancer pain or for the treatment of extremity disorders such as reducing intraocular pressure in glaucoma and dry eye and dry mouth such as in Sjogren's syndrome;

[0027] Use of the compounds of the present invention to treat skin lesions, such as those resulting from pemphigus vulgaris, dermatitis herpetiformis, pemphigoid and other blistering skin conditions; Use of the compounds of the present invention for treating, preventing, ameliorating or reversing conditions associated with altered gastrointestinal function and motility, such as functional dyspepsia, irritable bowel syndrome, gastroesophageal reflux (GER) and esophageal hypoperistalsis, gastroparesis and conditions of chronic diarrhea; Use of the compounds of the present invention to treat olfactory dysfunction such as Bosma-Henkin-Christiansen syndrome, chemical poisoning (e.g., selenium and silver), hypopituitarism, Kallmann syndrome, skull fractures, tumor treatment, and hypothyroidism; Use of the compounds of the present invention for treating addiction; Use of the compounds of the present invention for treating movement disorders such as Parkinson's disease, ADHD, Huntington's disease, Tourette's syndrome and other syndromes associated with dopaminergic dysfunction as the underlying pathogenic factor driving the disease; Use of the compounds of the present invention to treat behavioral and psychological symptoms of dementia (BPSD; including agitation, verbal aggression, physical aggression, depression, anxiety, abnormal motor behavior, elevated mood, irritability, blunted affect, disinhibition, impulsivity, delusions, hallucinations, sleep changes, and appetite changes).

[0028] Compounds of the present invention include Example 1, Example 1-1 and Example 1-2 shown below. [Table 1]

[0029] Methods for preparing the compounds of the present invention The compounds of the present invention can be prepared according to synthetic methods well known to those skilled in the art and described herein. Also provided is a process for preparing the compounds defined above, said process comprising any of A, B or C: (A) Formula (10): [ka] is converted under reductive amination conditions to a compound of formula (11): [ka] or (B) Formula (12): [ka] with an amine of formula (CH3)3CNH2, where R represents a suitable group such as methyl or ethyl; or (C) Formula (13): [ka] reacting the compound of formula (CH3)3CNH2 with an amine of formula (CH3)3CNH2.

[0030] Such methods are well known to those skilled in the art. Examples of synthetic procedures for converting one functional group into another are described in standard texts, such as March's Advanced Organic Chemistry: Reactions, Mechanisms, and Structure, 7th Edition, Michael B. Smith, John Wiley, 2013, (ISBN: 978-0-470-46259-1), Organic Syntheses, Online Edition, www.orgsyn.org, (ISSN 2333-3553), and Fiesers' Reagents for Organic Synthesis, Volumes 1-17, John Wiley, edited by Mary Fieser (ISBN: 0-471-58283-2). In the reactions described above, it may be necessary to protect one or more groups to prevent reaction from occurring at undesired locations on the molecule. Examples of protecting groups and methods for protecting and deprotecting functional groups are described in Greene's Protective Groups in Organic Synthesis, Fifth Edition, Editor: Peter GM Wuts, John Wiley, 2014, (ISBN: 9781118057483). The compounds prepared by the methods described above can be isolated and purified by any of a variety of methods well known to those skilled in the art, including recrystallization and chromatographic techniques such as column chromatography (e.g., flash chromatography), HPLC, and SFC.

[0031] Pharmaceutical preparations While it is possible for the active compound to be administered alone, it is preferable to present it as a pharmaceutical composition (eg, a formulation). Therefore, there is provided a pharmaceutical composition comprising at least one compound of the invention as defined above, together with at least one pharmaceutically acceptable excipient. The composition may be a tablet composition. The composition may be a capsule composition. The one or more pharmaceutically acceptable excipients can be selected from, for example, carriers (e.g., solid, liquid, or semi-solid carriers), adjuvants, diluents (e.g., solid diluents such as fillers or bulking agents; and liquid diluents such as solvents and co-solvents), granulating agents, binders, flow aids, coating agents, release-controlling agents (e.g., polymers or waxes that slow or retard release), binding agents, disintegrants, buffers, lubricants, preservatives, antifungal and antibacterial agents, antioxidants, buffers, tonicity-adjusting agents, thickeners, flavors, sweeteners, dyes, plasticizers, taste-masking agents, stabilizers, or any other excipient conventionally used in pharmaceutical compositions. As used herein, the term "pharmaceutically acceptable" means a compound, material, composition, and / or dosage form that is, within the scope of sound medical judgment, suitable for use in contact with the tissues of a subject (e.g., a human subject) without undue toxicity, irritation, allergic response, or other problem or complication, commensurate with a reasonable benefit / risk ratio. Each excipient must also be "acceptable" in the sense of being compatible with the other ingredients of the formulation. Pharmaceutical compositions containing the compounds of the present invention can be formulated according to known techniques, see, for example, Remington's Pharmaceutical Sciences, Mack Publishing Company, Easton, PA, USA. The pharmaceutical compositions may be in any form suitable for oral, parenteral, topical, intranasal, intrabronchial, sublingual, ophthalmic, optic, rectal, vaginal, or transdermal administration.

[0032] Pharmaceutical dosage forms suitable for oral administration include tablets (coated or uncoated), capsules (hard or soft shell), caplets, pills, lozenges, syrups, liquids, powders, granules, elixirs and suspensions, sublingual tablets, wafers or patches (e.g., buccal patches). Tablet compositions can contain a unit dose of active compound together with an inert diluent or carrier, such as a sugar or sugar alcohol (e.g., lactose, sucrose, sorbitol, or mannitol); and / or a non-sugar-based diluent, such as sodium carbonate, calcium phosphate, calcium carbonate, or a cellulose or its derivatives, such as microcrystalline cellulose (MCC), methylcellulose, ethylcellulose, hydroxypropylmethylcellulose, and starch, such as corn starch. Tablets can also contain standard ingredients, such as binders and granulating agents (e.g., polyvinylpyrrolidone), disintegrants (e.g., swellable cross-linked polymers such as cross-linked carboxymethylcellulose), lubricants (e.g., stearates), preservatives (e.g., parabens), antioxidants (e.g., BHT), buffers (e.g., phosphate buffers or citrate buffers), and effervescent agents (e.g., citrate / bicarbonate mixtures). Such excipients are well known and need not be discussed in detail herein. Tablets may be designed to release the drug upon contact with gastric fluids (immediate-release tablets) or may be designed to release in a controlled manner over an extended period of time or in specific areas of the GI tract (controlled-release tablets).

[0033] A pharmaceutical composition typically contains approximately 1% (w / w) to approximately 95% (preferably % (w / w)) of the active ingredient and 99% (w / w) to 5% (w / w) of a pharmaceutically acceptable excipient (e.g., as defined above) or a combination of excipients. Preferably, the composition contains approximately 20% (w / w) to approximately 90% (w / w) of the active ingredient and 80% (w / w) to 10% of a pharmaceutical excipient or a combination of excipients. A pharmaceutical composition contains approximately 1% to approximately 95%, preferably approximately 20% to approximately 90%, of the active ingredient. The pharmaceutical composition of the present invention may be in unit dose form, such as in the form of an ampule, vial, suppository, pre-filled syringe, dragee, powder, tablet, or capsule. Tablets and capsules may contain, for example, 0-20% disintegrant, 0-5% lubricant, 0-5% flow aid, and / or 0-99% (w / w) filler or bulking agent (depending on the drug dose). Tablets and capsules may also contain 0-10% (w / w) polymer binder, 0-5% (w / w) antioxidant, and 0-5% (w / w) pigment. Sustained-release tablets further typically contain 0-99% (w / w) release-controlling (e.g., retarding) polymer (depending on the dose). The film coat of a tablet or capsule typically contains 0-10% (w / w) polymer, 0-3% (w / w) pigment, and / or 0-2% (w / w) plasticizer. Parenteral formulations typically contain 0-20% (w / w) buffer, 0-50% (w / w) cosolvent, and / or 0-99% (w / w) water for injection (WFI) (depending on dose and whether lyophilized), and intramuscular depot formulations may contain 0-99% (w / w) oil.

[0034] Pharmaceutical formulations may be provided to the patient in "patient packs" that contain an entire course of treatment in a single package, usually a blister pack. The compounds of the invention are generally provided in unit dosage form and, thus, typically contain sufficient compound to provide a desired level of biological activity. For example, a formulation may contain 1 nanogram to 2 grams of active ingredient, e.g., 1 nanogram to 2 milligrams of active ingredient. Within these ranges, certain narrower ranges of compound are 0.1 milligram to 2 grams of active ingredient (more usually, 10 milligrams to 1 gram, e.g., 50 milligrams to 500 milligrams), or 1 microgram to 20 milligrams (e.g., 1 microgram to 10 milligrams, e.g., 0.1 milligram to 2 milligrams of active ingredient). For oral compositions, a unit dosage form may contain from 1 milligram to 2 grams, more typically 10 milligrams to 1 gram, for example, 50 milligrams to 1 gram, for example, 100 milligrams to 1 gram of active compound. The active compound is administered to a patient (e.g., a human or animal patient) in need thereof in an amount sufficient to achieve the desired therapeutic effect (an effective amount). The exact amount of compound to be administered can be determined by the attending physician according to standard procedures. [Example]

[0035] General Procedure Where no preparative route is described, the relevant intermediates are commercially available. Commercially available reagents were used without further purification. Room temperature (rt) refers to approximately 22-30°C. 1 H NMR spectra were recorded on a Bruker instrument at 400 MHz. Chemical shift values ​​are expressed in parts per million (ppm) (i.e., (δ) values). The following abbreviations are used for the multiplicity of NMR signals: s = singlet, br = broad, d = doublet, t = triplet, q = quartet, quint = quintet, td = triplet of doublets, tt = triplet of triplets, qd = quartet of doublets, ddd = doublet of doublet of doublets, ddt = doublet of doublet of triplets, m = multiplet. Coupling constants were reported as J values ​​measured in Hz. NMR and mass spectrometry results were corrected for background peaks. LCMS analysis LCMS analysis of compounds was carried out under electrospray conditions using the equipment and methods shown in the table below. [Table 2]

[0036] [Table 3]

[0037] Preparative HPLC purification Preparative HPLC purification was performed using a Shimadzu LC-20AP binary system equipped with an SPD-20A UV detector. Purification technique: [phase (column description, column length x inner diameter, particle size), solvent flow rate, gradient - mobile phase B in mobile phase A (over time) expressed as % mobile phase (A), mobile phase (B)]. Preparative HPLC method A Preparative HPLC: [reverse phase (X-BRIDGE C-18, 250 × 50 mm, 5 μm), 85 mL / min, gradient: 35% to 70% (26 min), 100% (2 min), 100% to 35% (6 min), mobile phase (A): 5 mM ammonium bicarbonate in water + 0.1% ammonia in water, (B): 100% acetonitrile] Preparative HPLC method B Preparative HPLC: [reverse phase (X-BRIDGE C-18, 250 × 50 mm, 5 μm), 85 mL / min, gradient: 40% to 60% (26 min), 60% (4 min), 100% (2 min), 100% to 40% (7 min), mobile phase (A): 5 mM ammonium bicarbonate in water + 0.1% ammonia in water, (B): 100% acetonitrile]

[0038] Abbreviation AcOH = acetic acid Bn = benzyl t-BuOH = tert-butyl alcohol CPM = cyclopentyl methyl ether DCM = dichloromethane DIPEA = N,N-diisopropylethylamine DMF = dimethylformamide DMSO = dimethyl sulfoxide ESI = electrospray ionization EtOH = ethanol h = time / second HATU = hexafluorophosphate azabenzotriazole tetramethyluronium HPLC = high-performance liquid chromatography IPA = propan-2-ol LCMS = liquid chromatography mass spectrometry MeOH = methanol min = minutes / seconds 2-MTHF = 2-methyltetrahydrofuran nm = nanometers NMO = 4-methylmorpholine 4-oxide NMR = nuclear magnetic resonance rt = room temperature RT = residence time TEA = triethylamine TFA = trifluoroacetic acid TFAA = trifluoroacetic anhydride THF = tetrahydrofuran The prefixes n-, s-, i-, t- and tert- have their usual meanings: normal, secondary, iso, and tertiary.

[0039] Synthesis of N-(tert-butyl)-1-((1R,3r,5S)-8-(3-(trifluoromethyl)-1,2,4-oxadiazol-5-yl)-8-azabicyclo[3.2.1]octan-3-yl)piperidine-4-carboxamide hydrochloride (Example 1-1) [ka] [ka]

[0040] Step 1: Synthesis of benzyl cyclopent-3-ene-1-carboxylate (Intermediate 2) To a mixture of K2CO3 (61.5 g, 446 mmol) and cyclopent-3-ene-1-carboxylic acid (CAS: 7686-77-3, Intermediate 1) (25.0 g, 223 mmol) in acetone (375 mL), (bromomethyl)benzene (CAS: 100-39-0) (29.1 mL, 245 mmol) was added dropwise. The reaction was stirred at 60 °C for 2 h and then cooled to room temperature. The resulting mixture was filtered, and the filtrate was concentrated under reduced pressure. The crude product was purified by column chromatography [normal phase (neutral Al2O3), 0–10% (ethyl acetate in hexane)] to give benzyl cyclopent-3-ene-1-carboxylate (Intermediate 2) (40.1 g, 89.0%). LCMS (System 1, Method A): (ESI) m / z 203 [M+H] + RT 5.10 min, 254 nm. [ka]

[0041] Step 2: Synthesis of benzyl 3,4-dihydroxycyclopentane-1-carboxylate (Intermediate 3) A mixture of 4-methylmorpholine 4-oxide (30.44 g, 260 mmol), OsO (2% in t-BuOH, 12.0 mL, 0.94 mmol), and benzyl cyclopent-3-ene-1-carboxylate (Intermediate 2) (43.8 g, 217 mmol) in acetone (431 mL) was stirred at room temperature for 16 h. The mixture was treated with saturated aqueous NaSO (500 mL) and then extracted with DCM (3 × 400 mL). The combined organic layers were dried (NaSO) and concentrated under reduced pressure. The resulting residue was purified by column chromatography [normal phase (silica), 0–50% (ethyl acetate in hexane)] to give benzyl 3,4-dihydroxycyclopentane-1-carboxylate (Intermediate 3) (30.4 g, 59.4%). LCMS (System 2, Method B): (ESI) m / z 237 [M+H] + RT 2.38 min, 224 nm. [ka]

[0042] Step 3: Synthesis of benzyl 4-oxo-2-(2-oxoethyl)butanoate (Intermediate 4) A mixture of NaIO (48.7 g, 229 mmol) and benzyl 3,4-dihydroxycyclopentane-1-carboxylate (Intermediate 3) (36.0 g, 153 mmol) in THF (1800 mL) and water (144 mL) was stirred at room temperature for 2 h. Water (1500 mL) was added until the precipitate dissolved, and then the mixture was extracted with DCM (3 × 500 mL). The combined organic layers were dried (NaSO) and concentrated under reduced pressure to give benzyl 4-oxo-2-(2-oxoethyl)butanoate (Intermediate 4) (35.8 g, 100.0%). The crude material was used without further purification. LCMS (System 2, Method B): (ESI) m / z 233 [M−H] - RT 2.33 min and 2.74 min, 202 nm. [ka]

[0043] Step 4: Synthesis of tert-butyl (1R,3r,5S)-3-(4-((benzyloxy)carbonyl)piperidin-1-yl)-8-azabicyclo[3.2.1]octane-8-carboxylate (Intermediate 6) A solution of benzyl 4-oxo-2-(2-oxoethyl)butanoate (Intermediate 4) (35.8 g, 153 mmol) and tert-butyl (1R,3r,5S)-3-amino-8-azabicyclo[3.2.1]octane-8-carboxylate (CAS: 207405-68-3, Intermediate 5) (34.6 g, 153 mmol) in EtOH (1400 mL) was stirred at room temperature for 30 min. NaBHCN (9.64 g, 153 mmol) and AcOH (3.0 mL, 52.5 mmol) were then added to the solution, and stirring was continued at room temperature for 16 h. The reaction mixture was diluted with water (1000 mL) and extracted with DCM (3 × 500 mL). The combined organic layers were dried (NaSO) and concentrated under reduced pressure. The resulting residue was purified by column chromatography [normal phase (silica), 0-30% (ethyl acetate in hexane)] to give tert-butyl (1R,3r,5S)-3-(4-((benzyloxy)carbonyl)piperidin-1-yl)-8-azabicyclo[3.2.1]octane-8-carboxylate (Intermediate 6) (41.0 g, 62.6%). LCMS (System 2, Method B): (ESI) m / z 429 [M+H] + RT 4.28 min, 202 nm. [ka]

[0044] Step 5: Synthesis of benzyl 1-((1R,3r,5S)-8-azabicyclo[3.2.1]octan-3-yl)piperidine-4-carboxylate dihydrochloride (Intermediate 7) To a solution of tert-butyl (1R,3r,5S)-3-(4-((benzyloxy)carbonyl)piperidin-1-yl)-8-azabicyclo[3.2.1]octane-8-carboxylate (Intermediate 6) (41.0 g, 96.0 mmol) in 1,4-dioxane (82.0 mL) was added HCl in 1,4-dioxane (4 M, 410 mL, 10 vol) dropwise at 0 °C. The reaction mixture was stirred at room temperature for 2 h and then concentrated under reduced pressure. The resulting residue was azeotroped with hexane (2 × 50 mL) and then triturated with hexane (2 × 50 mL) to give benzyl 1-((1R,3r,5S)-8-azabicyclo[3.2.1]octan-3-yl)piperidine-4-carboxylate dihydrochloride (Intermediate 7) (38.4 g, 100.0%). LCMS (System 1, Method B): (ESI) m / z 329 [M+H] + RT 3.10 min, 202 nm. [ka]

[0045] Step 6: Synthesis of benzyl 1-((1R,3r,5S)-8-cyano-8-azabicyclo[3.2.1]octan-3-yl)piperidine-4-carboxylate (Intermediate 8) To a solution of benzyl 1-((1R,3r,5S)-8-azabicyclo[3.2.1]octan-3-yl)piperidine-4-carboxylate dihydrochloride (Intermediate 7) (38.4 g, 96.0 mmol) in DCM (314 mL) was added triethylamine (39.8 mL, 287 mmol) dropwise at −20° C., maintaining the internal temperature in the range of −20° C. to 0° C. The reaction was then stirred at this temperature for 30 minutes. To the reaction was then added cyanogen bromide (15.1 g, 144 mmol) as a solution in DCM (31.0 mL), dropwise, maintaining the internal temperature in the range of −20° C. to 0° C. The reaction was then warmed to room temperature and stirred for 16 hours. The reaction mixture was diluted with saturated aqueous NaHCO (700 mL) and extracted with DCM (3×300 mL). The combined organic layers were dried (NaSO) and concentrated under reduced pressure. The crude material was purified by column chromatography [normal phase (neutral AlO), 0–30% (ethyl acetate in hexane)] to give benzyl 1-((1R,3r,5S)-8-cyano-8-azabicyclo[3.2.1]octan-3-yl)piperidine-4-carboxylate (Intermediate 8) (16.0 g, 47.4%). LCMS (System 2, Method B): (ESI) m / z 354 [M+H] + RT 3.69 min, 202 nm. [ka]

[0046] Step 7: Synthesis of benzyl 1-((1R,3r,5S)-8-((((tert-butoxycarbonyl)amino)oxy)(imino)methyl)-8-azabicyclo[3.2.1]octan-3-yl)piperidine-4-carboxylate (Intermediate 10) To a solution of benzyl 1-((1R,3r,5S)-8-cyano-8-azabicyclo[3.2.1]octan-3-yl)piperidine-4-carboxylate (Intermediate 8) (16.0 g, 45.3 mmol) in THF (160 mL) was added tert-butyl hydroxycarbamate (CAS: 36016-38-3, Intermediate 9) (6.63 g, 49.8 mmol) at 0 °C. The reaction mixture was stirred at 0 °C for 20 min. Zinc chloride in 2-MTHF (1.9 M, 47.7 mL, 90.6 mmol) was then added slowly to the solution at 0 °C, followed by stirring at room temperature for 16 h. The reaction mixture was quenched with water (300 mL) and extracted with ethyl acetate (3 × 400 mL). The combined organic layers were dried (NaSO) and concentrated under reduced pressure. The resulting crude material was triturated with hexane (2×50 mL) to give benzyl 1-((1R,3r,5S)-8-((((tert-butoxycarbonyl)amino)oxy)(imino)methyl)-8-azabicyclo[3.2.1]octan-3-yl)piperidine-4-carboxylate (Intermediate 10) (22.0 g, 100.0%). The crude material was used without further purification. LCMS (System 2, Method B): (ESI) m / z 487 [M+H] + RT 3.10 min, 202 nm. [ka]

[0047] Step 8: Synthesis of benzyl 1-((1R,3r,5S)-8-(3-(trifluoromethyl)-1,2,4-oxadiazol-5-yl)-8-azabicyclo[3.2.1]octan-3-yl)piperidine-4-carboxylate (Intermediate 11) To a solution of benzyl 1-((1R,3r,5S)-8-((((tert-butoxycarbonyl)amino)oxy)(imino)methyl)-8-azabicyclo[3.2.1]octan-3-yl)piperidine-4-carboxylate (Intermediate 10) (22.0 g, 5.3 mmol) in DCM (220 mL) was added TFA (110 mL, 5 vol) dropwise at 0°C-5°C. The reaction mixture was stirred at room temperature for 40 minutes and then cooled to 0°C-5°C. To this was then added TFAA (28.7 mL, 203.7 mmol) dropwise and stirred for 30 minutes before warming to room temperature and stirring for 16 hours. The reaction mixture was diluted with toluene (220 mL) and concentrated under reduced pressure. Saturated NaHCO3 solution (800 mL) was added and the reaction was extracted with ethyl acetate (3 x 500 mL). The combined organic layers were dried (NaSO) and concentrated under reduced pressure. The resulting residue was purified by column chromatography [normal phase (neutral AlO), 0–30% (ethyl acetate in hexane)] to give benzyl 1-((1R,3r,5S)-8-(3-(trifluoromethyl)-1,2,4-oxadiazol-5-yl)-8-azabicyclo[3.2.1]octan-3-yl)piperidine-4-carboxylate (Intermediate 11) (12.4 g, 59.0%). LCMS (System 2, Method B): (ESI) m / z 465 [M+H] + RT 4.40 min, 240 nm. [ka]

[0048] Step 9: Synthesis of 1-((1R,3r,5S)-8-(3-(trifluoromethyl)-1,2,4-oxadiazol-5-yl)-8-azabicyclo[3.2.1]octan-3-yl)piperidine-4-carboxylic acid hydrochloride (Intermediate 12) To a solution of benzyl 1-((1R,3r,5S)-8-(3-(trifluoromethyl)-1,2,4-oxadiazol-5-yl)-8-azabicyclo[3.2.1]octan-3-yl)piperidine-4-carboxylate (Intermediate 11) (12.3 g, 26.5 mmol) in THF (123 mL) and water (24.6 mL), lithium hydroxide monohydrate (2.78 g, 66.3 mmol) was added portionwise at room temperature, followed by stirring at room temperature for 16 hours. The reaction mixture was diluted with water (100 mL), cooled to 0°C-10°C, and the pH was adjusted to 5-6 using 1 M HCl solution (approximately 70-80 mL). The reaction mixture was stirred for 1 hour, then the aqueous layer was separated and lyophilized to give 1-((1R,3r,5S)-8-(3-(trifluoromethyl)-1,2,4-oxadiazol-5-yl)-8-azabicyclo[3.2.1]octan-3-yl)piperidine-4-carboxylic acid hydrochloride (Intermediate 12) (15.1 g, crude), which was used without further purification. LCMS (System 2, Method B): (ESI) m / z 375 [M+H] + RT 2.10 min, 236 nm. [ka]

[0049] Step 10: Synthesis of N-(tert-butyl)-1-((1R,3r,5S)-8-(3-(trifluoromethyl)-1,2,4-oxadiazol-5-yl)-8-azabicyclo[3.2.1]octan-3-yl)piperidine-4-carboxamide (Intermediate 14) To a solution of 1-((1R,3r,5S)-8-(3-(trifluoromethyl)-1,2,4-oxadiazol-5-yl)-8-azabicyclo[3.2.1]octan-3-yl)piperidine-4-carboxylic acid hydrochloride (Intermediate 12) (5.8 g, 14.1 mmol) (8 g of crude material isolated from Step 9 was used) in DMF (80.0 mL) was added HATU (12.2 g, 32.0 mmol) in portions at 0° C. to 10° C. The reaction was stirred at 0° C. to 10° C. for 40 minutes. To this reaction was then added 2-methylpropan-2-amine (CAS: 75-64-9, Intermediate 13) (6.8 mL, 64.2 mmol) and DIPEA (11.4 mL, 64.2 mmol). The resulting reaction mixture was allowed to warm to room temperature and stirred for 16 hours. Ice water (500 mL) was added, stirred for 20 minutes, and the precipitate that formed was collected by filtration. The filter cake was washed with cold water (500 mL) and then with hexane (500 mL) to give crude product (2.5 g). The aqueous layer was extracted with ethyl acetate (2 × 200 mL), and the combined organic layers were dried (NaSO) and concentrated under reduced pressure to give additional crude product (4.5 g). The combined crude product was purified by column chromatography [normal phase (silica), 0–50% (ethyl acetate in hexane)] to give two batches of product with different purities (2.0 g and 3.8 g). Each batch was then removed separately and further purified by preparative HPLC methods A and B, respectively. The resulting products were combined and crystallized using IPA (10 vol) and MeOH (1 vol), and the filter cake was washed with cold IPA (2 vol) to give N-(tert-butyl)-1-((1R,3r,5S)-8-(3-(trifluoromethyl)-1,2,4-oxadiazol-5-yl)-8-azabicyclo[3.2.1]octan-3-yl)piperidine-4-carboxamide (Intermediate 14) (1.9 g, 31.4%). LCMS (System 2, Method B): (ESI) m / z 430 [M+H] + RT 3.54 min, 240 nm. 1H NMR (400 MHz, DMSO-d6): δ 7.31 (br. s, 1H), 4.39 - 4.25 (m, 2H), 3.26 - 3.15 (m, 2H), 2.27 - 1.80 (m, 10H), 1.71 - 1.44 (m, 6H), 1.22 (s, 9H). [ka]

[0050] Step 11: Synthesis of N-(tert-butyl)-1-((1R,3r,5S)-8-(3-(trifluoromethyl)-1,2,4-oxadiazol-5-yl)-8-azabicyclo[3.2.1]octan-3-yl)piperidine-4-carboxamide hydrochloride (Example 1-1) To a solution of N-(tert-butyl)-1-((1R,3r,5S)-8-(3-(trifluoromethyl)-1,2,4-oxadiazol-5-yl)-8-azabicyclo[3.2.1]octan-3-yl)piperidine-4-carboxamide (Intermediate 14) (1.9 g, 4.4 mmol) in 1,4-dioxane (3.8 mL) was added HCl in 1,4-dioxane (4 M, 19.0 mL, 10 vol) at 0° C. The reaction was stirred at room temperature for 4 hours and then concentrated under reduced pressure. The resulting residue was azeotroped with 1,4-dioxane (2×10 mL) and then triturated with 1,4-dioxane (10 mL). The solid was collected by filtration, and the filter cake was washed with 1,4-dioxane (2 × 5 mL), n-pentane (10 mL), and dried under reduced pressure. The solid was crystallized using IPA (10 vol) and MeOH (1 vol), and the filter cake was washed with cold IPA (2 vol) to give N-(tert-butyl)-1-((1R,3r,5S)-8-(3-(trifluoromethyl)-1,2,4-oxadiazol-5-yl)-8-azabicyclo[3.2.1]octan-3-yl)piperidine-4-carboxamide hydrochloride (Example 1-1) (1.8 g, 87.9%). LCMS (System 2, Method B): (ESI) m / z 430 [M+H] + RT 3.55 min, 240 nm. 1H NMR (400 MHz, DMSO-d6): δ 10.78 - 10.24 (m, 1H), 7.58 - 7.46 (m, 1H), 4.62 - 4.50 (m, 2H), 3.54 - 3.43 (m, 2H), 3.32 - 3.06 (m, 2H), 2.85 - 2.64 (m, 4H), 2.37 - 2.20 (m, 1H), 2.13 - 2.00 (m, 2H), 1.93 - 1.69 (m, 7H), 1.23 (s, 9H).

[0051] Scale-up synthesis of N-(tert-butyl)-1-((1R,3r,5S)-8-(3-(trifluoromethyl)-1,2,4-oxadiazol-5-yl)-8-azabicyclo[3.2.1]octan-3-yl)piperidine-4-carboxamide monohydrochloride monohydrate (Example 1-2)

change

[0052] Step 1: Synthesis of benzyl cyclopent-3-ene-1-carboxylate (2)

change

[0053] Step 2: Synthesis of benzyl 6-oxabicyclo[3.1.0]hexane-3-carboxylate (15) [ka] A clean, dry 5.0-L four-neck RB flask equipped with a condenser and a thermometer pocket was charged with MTBE (2450 mL) and benzyl cyclopent-3-ene-1-carboxylate (Intermediate 2, 245 g) at room temperature (22–25°C) under a nitrogen atmosphere. The reaction mixture was cooled to 5–10°C. m-CPBA (362.6 g) was added in five separate lots, causing the color to change from dark brown to yellow. The reaction mixture was slowly warmed to 15–20°C and stirred at 15–20°C for 16–20 hours. The reaction progress was monitored by HPLC until completion. 20% aqueous sodium bisulfite solution (3675 mL, 15V) was added slowly over 15–20 minutes, followed by stirring at 20–25°C for 30 minutes. The organic layer was separated, and 10% aqueous Na2CO3 (2450 mL, 10V) was added and stirred at 20-25°C for 15 minutes. The organic layer was separated, and 20% aqueous sodium bisulfite (2450 mL, 10V) was added and stirred until the peroxide content was <3 mg / L (peroxide strip). The organic layer was separated, dried over sodium sulfate, filtered, and evaporated under pressure at 37-40°C to give benzyl 6-oxabicyclo[3.1.0]hexane-3-carboxylate. Crude mass: 234g, Crude yield: 88%, Crude characteristics: pale yellow liquid The crude material was used directly in the next step without further purification.

[0054] Step 03: Synthesis of benzyl 4-oxo-2-(2-oxoethyl)butanoate (4) [ka] A clean, dry 2.0 L four-neck RB flask equipped with a condenser and a thermometer pocket was charged with ethyl acetate (380 mL) and periodic acid (87.41 g) at room temperature (22–25 °C) under a nitrogen atmosphere, and the resulting white suspension was cooled to 0–10 °C. Upon addition of benzyl 6-oxabicyclo[3.1.0]hexane-3-carboxylate (Intermediate 15, 76 g in 380 mL of ethyl acetate), a color change from dark brown to yellow was observed. The reaction mixture was slowly warmed to 15–20 °C and stirred for 3–4 h. The reaction progress was monitored by TLC and HPLC until completion. Demineralized water (760 mL, 10 V) was added and stirred at 20–25 °C for 15–20 min, three times. The organic layer was separated, washed with brine solution (380 mL, 5 V), and stirred at 20–25 °C for 5–10 min. The organic layer was separated, dried over sodium sulfate, filtered, and the organic layer was evaporated under pressure at 37-40° C. to give benzyl-4-oxo-2-(2-oxoethyl)butanoate. Mass of crude product: 82g Characteristics of crude product: pale yellow liquid The crude material was used directly in the next step without further purification.

[0055] Step 04: Synthesis of tert-butyl (1R,3R,5S)-3-(4-((benzyloxy)carbonyl)piperidin-1-yl)-8-azabicyclo[3.2.1]octane-8-carboxylate (6) [ka] A clean, dry 2.0 L four-neck RB flask equipped with a condenser and thermometer pocket was charged with 2-methyl-THF (570 mL) and benzyl-4-oxo-2-(2-oxoethyl)butanoate (Intermediate 4, 57 g (actual crude mass obtained directly from Step 3 was 82 g) at room temperature (22-23 °C) under a nitrogen atmosphere. Intermediate 5 (endo amine) (49.54 g) was added and the reaction was cooled to 5-10 °C and the reaction was stirred for 30 minutes. Sodium triacetoxyborohydride (56.78 g) and glacial acetic acid (5.7 g) were added. (mL) was added, and the reaction mixture was warmed to room temperature (22-23°C) and stirred for 10-12 hours. The reaction progress was monitored by TLC and HPLC. Upon completion, saturated bicarbonate solution (400 mL, 7V) was added and stirred at 20-25°C for 15-20 minutes. The organic layer was separated, dried over sodium sulfate, filtered, and evaporated under reduced pressure at 37-40°C to give tert-butyl (1R,3R,5S)-3-(4-((benzyloxy)carbonyl)piperidin-1-yl)-8-azabicyclo[3.2.1]octane-8-carboxylate. Mass of crude product: 115g, Characteristics of crude product: Thick liquid, Color: Light chestnut

[0056] Purification with oxalate formation: A clean, dry 500 mL four-neck RB flask equipped with a thermometer and condenser was charged with the crude product (25 g) and acetone (150 mL, 6 V) and cooled to 5–10°C. Oxalic acid (10 g, 2.0 eq.) was added, and the reaction mass was allowed to warm to room temperature with stirring for 2 h. The reaction mass was evaporated at 40°C to give the crude product (mass: 35 g), which was then added with 10 Vol of MTBE at 25–30°C and stirred for 30 min. The reaction mass was filtered and a bed wash with MTBE (1.0 vol) gave a wet solid (oxalate salt, 26 g). The solid was suspended in saturated bicarbonate solution (30 vol, 780 mL) and ethyl acetate (25 Vol, 650 mL) and stirred for 15 min. The organic layer was separated, dried over Na2SO4, and evaporated under vacuum at 37–40°C to give the product. Mass of product: 8.0 g If necessary, this material can be further purified by column chromatography by adsorption onto neutral alumina (Source - SDFCL) using hexane:ethyl acetate. Column gradient: 5->10->12% (ethyl acetate:hexane).

[0057] Step 05: Synthesis of benzyl 1-((1R,3R,5S)-8-azabicyclo[3.2.1]octan-3-yl)piperidine-4-carboxylate hydrochloride (7) [ka] A clean, dry 5.0-liter four-neck RB flask equipped with a condenser and thermometer pocket was charged with cyclopentyl methyl ether (452 ​​mL, 4.0 V) and Intermediate 6 (113 g, 1.0 eq.) under a nitrogen atmosphere, and the reaction mixture was cooled to 0-5°C. 3 M HCl in cyclopentyl methyl ether (904 mL, 8.0 V) was added very slowly at 0-5°C, and the reaction mixture was slowly warmed to 22-25°C and stirred for 16 h. The reaction was monitored by HPLC until completion. The reaction mixture was filtered under a N2 atmosphere, and a bed wash was performed with MTBE (2.0 vol.). The product wet cake was removed under N2 and dried under reduced pressure at 45-47°C to give the crude product, which was used directly in the next step without any further purification.

[0058] Step 06: Synthesis of benzyl 1-((1R,3R,5S)-8-cyano-8-azabicyclo[3.2.1]octan-3-yl)piperidine-4-carboxylate (8) [ka] A clean, dry 5.0-L four-neck RB flask equipped with a condenser and thermometer pocket was charged with dichloromethane (1860 mL, 10 vol.), Intermediate 7 (186.0 g, 1.0 eq.), and triethylamine (355 mL, 5.0 eq.) over 10–15 min at 22–25°C under a nitrogen atmosphere. The reaction mixture was stirred for 30–45 min. The reaction mixture was cooled to 0–5°C, and cyanogen bromide (92 g, 1.7 eq.) in DCM (372 mL, 2.0 V) was added at 0–5°C. The reaction mixture was slowly warmed to 22–25°C and stirred at 22–25°C for 3–4 h. The reaction progress was monitored by HPLC until completion. The reaction mixture was diluted with DCM (1860 mL, 10 vol.) and basified with saturated NaHCO3 solution (930 mL, 5 V). The organic layer was separated and the aqueous layer was extracted with DCM (1860 mL, 10 vol). The combined organic layers were dried over Na2SO4 and evaporated under reduced pressure to give 190 g of crude compound. The crude compound was purified by neutral alumina column chromatography using 15% ethyl acetate in hexane. The pure fractions were collected and concentrated under reduced pressure to give 75 g of pure compound (yield: 46%).

[0059] Step 07: Synthesis of benzyl 1-((1R,3R,5S)-8-((((tertbutoxycarbonyl)amino)oxy)(imino)methyl)-8-azabicyclo[3.2.1]octan-3-yl)piperidine-4-carboxylate (10) [ka] A clean, dry 3.0 L four-neck RB flask equipped with a condenser and thermometer pocket was charged with 2-methyl-THF (800 mL, 10 vol.) and Intermediate 8 (80 g, 1.0 eq.) under a nitrogen atmosphere, and the reaction mixture was cooled to 0–5°C. A solution of ZnCl (2.2 eq., 1.9 M solution in 2-methyl-THF) was added, and the reaction mixture was stirred at 0°C for 30 min. Next, N-Boc-hydroxylamine (Intermediate 9) (1.2 eq.) was added, and the reaction mixture was slowly warmed to 23–25°C and continued stirring for 16 h. The reaction progress was monitored by TLC and HPLC until completion. The reaction mixture was quenched with water (800 mL, 10.0 V), and the product was then extracted with ethyl acetate (2 × 800 mL). The combined organic layers were dried over NaSO and evaporated under reduced pressure to give 110 g (quantitative yield) of crude compound. This product was used directly in the next step without further purification.

[0060] Step 08: Synthesis of benzyl 1-((1R,3R,5S)-8-(3-(trifluoromethyl)-1,2,4-oxadiazol-5-yl)-8-azabicyclo[3.2.1]octan-3-yl)piperidine-4-carboxylate (11) [ka] A clean, dry 3.0 L four-neck RB flask equipped with a condenser and thermometer pocket was charged with DCM (1150 mL, 10.0 vol.) and Intermediate 10 (115.0 g, 1.0 eq.) under a nitrogen atmosphere, and the reaction mixture was cooled to 0-5°C. A mixture of trifluoroacetic acid (271 mL, 15.0 eq.) and TFAA (164 mL, 5.0 eq.) was added dropwise in three portions at 3-4 h intervals under a nitrogen atmosphere at 0-5°C. The reaction mixture was stirred at room temperature for 12 h and then for an additional 16 h until the reaction was complete (monitored by TLC and HPLC). After completion of the reaction, the reaction mixture was cooled to 0°C, the pH was adjusted to 7-8 with saturated aqueous NaHCO3, and the reaction mixture was stirred for 15 min. The layers were separated, and the aqueous layer was extracted with DCM (2 × 100 mL). The combined organic layers were dried over NaSO and evaporated under reduced pressure to give 145 g of crude compound, which was purified by column chromatography on neutral alumina using hexane and ethyl acetate as eluents to give 49 g of pure product.

[0061] Step 09: Synthesis of 1-((1R,3R,5S)-8-(3-(trifluoromethyl)-1,2,4-oxadiazol-5-yl)-8-azabicyclo[3.2.1]octan-3-yl)piperidine-4-carboxylic acid (12) [ka] A clean, dry 5.0-liter four-neck RB flask equipped with a condenser and a thermometer pocket was charged with THF (2250 mL), demineralized water (450 mL), and Intermediate 11 (225 g) at room temperature (22-25 °C) under a nitrogen atmosphere. Lithium hydroxide monohydrate (30.5 g, 2.0 eq.) was added at room temperature (22-25 °C), and the reaction mixture was stirred at 60 °C for 4 h. The reaction progress was monitored by TLC and HPLC. Upon completion of the reaction, the reaction mixture was cooled to 0-5 °C. The pH was adjusted to 4-5 with 1 N HCl solution, and the mixture was stirred for 15 min. The solvent was evaporated, and the crude solid compound was purified by adding MTBE (10.0 V) to the crude solid compound at room temperature and stirring for 2 h. The solid was filtered and washed with MTBE (10.0 V). The solid compound was dried under vacuum to give 245 g of product as an off-white solid (216 g of the actual compound was Intermediate 12, the remaining approximately 29.0 g was the lithium chloride salt).

[0062] Step 10: Synthesis of N-(tert-butyl)-1-((1R,3r,5S)-8-(3-(trifluoromethyl)-1,2,4-oxadiazol-5-yl)-8-azabicyclo[3.2.1]octan-3-yl)piperidine-4-carboxamide [ka] procedure: A clean, dry 1 L three-necked round-bottom flask equipped with a mechanical stirrer and nitrogen inlet was charged with acetonitrile (20.0 V) and Intermediate 12 (1.0 eq) in a single lot, and the reaction mixture was stirred for 10 min. The reaction mixture was cooled to 0 °C, and then HATU was added, followed by the slow addition of tert-butylamine (1.5 eq) and DIPEA (4.0 V) over 15 min. After the addition was complete, the reaction mixture was stirred at room temperature for 16 h. The reaction progress was monitored by TLC, and upon completion, excess acetonitrile was distilled off under reduced pressure. The reaction was quenched with demineralized water (40.0 V), and the solid material precipitated and stirred for 60 min. The solid was washed with demineralized water (5.0 V) and dried by suction. The solid compound was dried under vacuum to give 56.0 g of the product as an off-white solid.

[0063] Step 11: Synthesis of N-(tert-butyl)-1-((1R,3r,5S)-8-(3-(trifluoromethyl)-1,2,4-oxadiazol-5-yl)-8-azabicyclo[3.2.1]octan-3-yl)piperidine-4-carboxamide hydrochloride [ka] A flask was charged with the free base of intermediate 14 (56.0 g, 1.0 eq.) in acetone (10.0 V), and the reaction mixture was stirred for 10 minutes. The reaction mixture was then cooled to 0°C, and a 15% HCl solution in IPA was added dropwise at 0°C. The reaction mixture was then warmed to 25-30°C and stirred at 25-30°C for 2 hours. The solvent was completely evaporated under reduced pressure and co-evaporated twice with n-heptane at 10.0 V. The solid compound was dried under vacuum to give 59.0 g of the product as an off-white solid.

[0064] Step 12 (Recrystallization): Synthesis of N-(tert-butyl)-1-((1R,3r,5S)-8-(3-(trifluoromethyl)-1,2,4-oxadiazol-5-yl)-8-azabicyclo[3.2.1]octan-3-yl)piperidine-4-carboxamide monohydrochloride monohydrate (Example 1-2) A flask was charged with the hydrochloride salt of intermediate 14 (108.0 g) in IPA (10.0 V), and the reaction mixture was heated to reflux for 1 h; however, the solid did not completely dissolve at reflux. While continuing to heat for 1 h, MeOH (1.0 V) was added to obtain a solution. The hot solution was passed through a filter paper to remove insoluble particles. The solvent (approximately 8 volumes of the 11 volumes of solvent) was removed under reduced pressure. Approximately 3 volumes remained, which was cooled to 0 °C and stirred at the same temperature for 1 h. The solid was then filtered, washed with a minimum amount of chilled IPA, and dried by suction. The solid compound was dried under vacuum to obtain 93.5 g of the product as an off-white solid. Analysis was performed to determine the composition of the batch, which was found to be the monohydrochloride monohydrate. Monohydrochloride monohydrate formula: C 20 H 33 N5O3ClF3; molecular weight: 483.96 [Table 4]

[0065] biological activity Example A: Phosphorylation-ERK1 / 2 Assay Functional assays were performed using the Alphascreen Surefire Phospho-ERK1 / 2 Assay (Crouch & Osmond, Comb. Chem. High Throughput Screen, 2008). ERK1 / 2 phosphorylation is a downstream consequence of Gq / 11 and G i / o protein-coupled receptor activation, and is much more suitable for assessing M1, M3 receptors (Gq / 11-coupled) and M2, M4 receptors (G i / o-coupled) than using different assay formats for different receptor subtypes. CHO cells stably expressing human muscarinic M1, M2, M3, or M4 receptors were plated (25K / well) in 96-well tissue culture plates in MEM-α + 10% dialyzed FBS. Once attached, the cells were serum-starved overnight. Agonist stimulation was performed by adding 5 μL of agonist to the cells for 5 minutes at 37°C. The medium was removed, and 50 μL of lysis buffer was added. After 15 minutes, 4 μL of sample was transferred to a 384-well plate and 7 μL of detection mixture was added. The plate was incubated in the dark with gentle shaking for 2 hours and then read on a PHERAstar plate reader. Data obtained for each receptor subtype were used to determine pEC 50 and E max The results are shown in Table 1 below. [Table 5]

[0066] Example B: CLint (in vitro hepatocytes) (Example 1) Hepatocyte stability assays were performed using pooled cryopreserved hepatocytes (Bioreclamation). Test compounds prepared in DMSO were incubated at an initial concentration of 1 μM (final: 0.25% DMSO, n = 2) with hepatocytes at a cell density of 1 million cells / mL at 37 °C. Aliquots were removed at 0.5, 5, 10, 15, 30, 60, and 120 min, the reaction was stopped, and the compound was extracted with acetonitrile containing an analytical internal standard (0.5 μM carbamazepine). Samples were centrifuged, and the supernatant fractions were analyzed for the parent compound by mass spectrometry (LC-MS / MS). The amount of remaining compound (expressed as %) was determined from the MS response of each sample compared to that of the T = 0 sample (normalized to the internal standard). Ln plots of the remaining % were used to determine the half-life of compound elimination using the following relationship: Half-life (min) = -0.693 / λ, where λ is the slope of the Ln% remaining vs. time curve. formula: CLint (µL / min / million cells) = (0.693 / half-life (min)) x (1000 / million cells per mL of incubation) was used to calculate the in vitro intrinsic clearance (CLint) as μL / min / million cells. Mouse = 7 μL / min / million Rat: 8 μL / min / 1 million Dog 8μL / min / 1 million Monkeys <5μL / min / million Human <5μL / min / million

[0067] Example C: MDCK Permeability / Efflux (Example 1) MDR1-MDCK cells (Solvo Biotechnology) were cultured at 2.35 × 10 per well. 5Cells were seeded onto 24-well Transwell plates and cultured at 37°C under 5% CO2 for 3 days before use as confluent monolayers. For both apical-to-basolateral (A to B) and basolateral-to-apical (B to A) measurements, test compounds and control compounds (propranolol, vinblastine) were added (1 μM, final: 0.1% DMSO, n=2) to the donor compartment of the Transwell plate in assay buffer (Hank's balanced salt solution supplemented with 25 mM HEPES and adjusted to pH 7.4) for each cell type. Incubations were performed at 37°C, and samples were removed from both the donor and acceptor chambers at T=0 and 1 h. Compounds were analyzed by mass spectrometry (LC-MS / MS) with an internal standard for analysis. The following relation: Papp = (Acceptor compound at end of T / (Donor compound x Donor V) / Incubation time) x Area of ​​Donor V x 60 x 10 -6 cm / s From this, the apparent permeability (Papp) value was determined.

[0068] where V is the volume of each Transwell compartment (125 μL apical, 600 μL basolateral), Concentration is the relative MS response for the compound (normalized to an internal standard) in the donor chamber before incubation and in the acceptor chamber at the end of incubation, and Area is the area of ​​the cell exposed to drug translocation (0.33 cm 2 ). The efflux ratio (Papp B to A / Papp A to B) was calculated from the average Papp values ​​in each direction. The MDR1-MDCK cell line was engineered to overexpress the efflux transporter MDR1 (P-glycoprotein). The finding of good permeability from B to A but poor permeability from A to B suggests that the compound is a substrate for this transporter. Lucifer Yellow (LY) was added to the apical buffer in all wells to assess cell layer viability. High levels of LY transport indicate poor cell layer integrity, as LY cannot freely permeate the lipophilic barrier, and LY Papp >10 × 10 -6Wells with a flow rate of less than 1000 cm / s were rejected. Note that compromised integrity in one well does not affect the validity of other wells on the plate. Compound recovery from a well was determined from the MS response (normalized to an internal standard) in the donor and acceptor chambers at the end of incubation compared to the response in the donor chamber pre-incubation. Recovery <50% may suggest problems with compound solubility, stability, or binding in the assay, reducing the reliability of the results. AB=66×10 -6 cm / sec BA=77×10 -6 cm / sec BA / AB emission ratio=1.2

[0069] Example D: Solubility Data (Example 1) Water-soluble (thermodynamic)-LCMS / MS method A 10 mM stock solution (in DMSO) of the test sample was prepared. A 1 μM working solution was prepared by diluting the test sample from the 10 mM stock solution with mobile phase solution (typically methanol: 2 mM ammonium acetate containing the appropriate internal standard (IS), carbamazepine / other appropriate IS). The working solution was also serially diluted with mobile phase solution (up to 5–6 linearity points) to prepare standard solutions for plotting calibration curves. The area for each standard sample was analyzed using LCMS / MS. The normalized area values ​​were plotted against concentration to obtain a calibration equation for determining unknown samples. To confirm the hydrothermodynamic (TD) solubility of the test compound, 1 mg of the compound (powdered) was added to 1 mL of each buffer and biorelevant medium listed in the table below to achieve a theoretical concentration equivalent to 1 mg / mL. The test compound was dispersed in the buffer solution using a vortex mixer.

[0070] [Table 6] The resulting solution was then placed on a RotoSpin shaker at 50 rpm for 4 hours for TD solubility at room temperature (25°C). After the incubation period, the solution was filtered using a 0.45 micron PVDF syringe filter to remove the insoluble fraction of the compound. The filtrate was diluted with mobile phase, and the AUC of the diluted sample was confirmed using LCMS / MS. The corresponding concentrations were calculated from the AUC of the test sample using a 5- to 6-point linearity / calibration curve. All numbers are reported as μM. [Table 7]

[0071] Example E: HμREL (Example 1) HμREL human Pool™ 96-well hepatocyte co-culture plate (HμREL human Pool TM Upon arrival of the 96-well hepatic co-culture plates, the medium was immediately changed and the cells were allowed to acclimate at 37°C for approximately 20 hours. The reaction was initiated by adding HμREL® incubation medium (serum-free) and test compounds (final substrate concentration: 1 μM; final DMSO concentration: 0.1%) to the HμREL® 96-well co-culture system (final cell number: 30,000 cells per well). The final incubation volume was 80 μL per time point. Two control compounds were included per assay. All incubations were performed alone for each test compound. Each compound was incubated for 0, 2, 6, 24, 48, and 72 hours (0, 120, 360, 1440, 2880, and 4320 minutes). At the appropriate time points, the reaction was stopped by transferring 60 μL of the incubation to 180 μL of acetonitrile containing the internal standard. The termination plate was centrifuged at 3000 rpm for 20 minutes at 4°C to precipitate any residual protein. quantitative analysis After protein precipitation, sample supernatants were pooled into cassettes of up to four compounds and analyzed using Cyprotex generic LC-MS / MS conditions. Data analysis The slope of the line was determined from a plot of the ln peak area ratio (compound peak area / internal standard peak area) versus time. The following equation was then used: Elimination rate constant (k) = (-slope) TIFF0007730369000043.tif10166 TIFF0007730369000044.tif10166 (where V = incubation volume (μL) / number of cells) CL int <0.143μL / min / million Using the half-life (t 1 / 2 ) and intrinsic clearance (CL int ) was calculated.

[0072] Example F: Predicted Target Engagement for Human Effective Dose (Example 1) The expected requirement for observing efficacy of M1 agonists in humans is the EC of recombinant M1. 50 Example 1 is predicted to achieve the unbound brain exposure required for M1 agonist efficacy at approximately a 22 mg dose, with a predicted human half-life of 15 hours (Figure 1). Parameters used to predict receptor occupancy (target engagement) MW=429.48 M1pEC 50 =7.17 Fu=0.682 Kpuu=1 Half-life (estimated) = 15 hours F=0.61 Cl=4.4mL / min / kg V = 5.9 mL / min / kg Ka=1 Definition: fu - fraction of unbound compound in plasma or brain tissue homogenate F-bioavailability; % of dose that reaches systemic circulation (plasma) Kp,uu - Ratio of unbound brain concentration / unbound plasma concentration. Quantifying the net flux of a drug across the blood-brain barrier (unconfounded by nonspecific binding in plasma and brain tissue, including the quantitative role of transporters). Gupta et al, DMD, 2006; Hammarlund-Udenaes et al, PharmRes, 2008

[0073] Example G: Attenuation of subchronic PCP-induced deficits in an operant reversal learning task in female Lister hooded rats (Example 1) Objectives and Results The ability of Example 1 (1, 3, 10 and 30 mg / kg, po, 1 hour pretreatment time, ptt) to attenuate disruptions in cognitive tasks induced by subchronic treatment with phencyclidine (scPCP) in female Lister hooded rats was investigated. Compared with vehicle, the percentage of correct responses in the reversal phase of the task was significantly (P<0.01) decreased in the scPCP group (Figure 2). In the reversal phase, treatment with Example 1 at the lowest dose and two intermediate doses (1, 3, and 10 mg / kg) significantly increased the percentage of correct responses (P<0.05, P<0.05, and P<0.01, respectively) compared with the scPCP group (Figure 2). Materials and Methods Female Lister hooded rats were used in this study. The average rat weight at the time of testing was 294 g ± 29 g. Rats were housed in groups of 3-5 under standard laboratory conditions with a 12-h light-dark cycle (lights on at 7:00 AM) and food was restricted to 90% of their free-feeding body weight (12 g of food per rat per day). Testing was performed during the light phase. Rats were randomly assigned to two treatment groups and received either vehicle, n = 8 (0.9% saline solution, i.p.), or PCP, n = 48 (2 mg / kg, i.p., twice daily for 7 days). On the day of testing, rats were randomly assigned to seven treatment groups (n = 6-8 per group) and received acute treatment with Example 1 (1, 3, 10, and 30 mg / kg, po, 1 h, ptt) or vehicle. Example 1 was dissolved in 1% methylcellulose and administered orally (po) at a volume of 5 mL / kg 1 hour before the test. The test was conducted in accordance with the Animals Scientific Procedures Act (UK, 1986) and was approved by AWERB (Animal Welfare and Ethical Review Body) of the University of Manchester.

[0074] Test Procedure After acclimation to the operant chamber, rats were trained to respond for food on a fixed ratio 1 (FR1) schedule of reinforcement with both levers active. When responding stabilized, rats were trained to press either the left or right lever for food, with the active lever alternating between days. Each session lasted 20 min, and counts were recorded for each lever. Rats were then further trained to respond for food based on the location of a visual cue (illuminated LED). Half were trained to press the lever under the illuminated LED to receive a food reward, while the other half were trained under the reversed contingency (lever pressing under the unilluminated LED). The test session ended after a total of 128 lever presses and lasted approximately 30 min. Rats were then retrained until they reached criterion on the reversed contingency. The day before each reversal learning task session, a full 30-minute operant training session (as described above) was conducted to ensure stable responding. For the reversal learning task, animals were first exposed to a 5-minute period during which the contingency (location of the cue relative to the active lever) was the same as in the operant training session. During this period, responses on both the correct and incorrect levers were recorded. This portion of the session is referred to as the initial phase. During the next 5 minutes, the contingency was reversed. Responses on the correct and incorrect levers were again recorded. This second period is referred to as the reversal phase. At this stage, training was terminated, and rats were treated with PCP (2 mg / kg, i.p.) or vehicle (0.9% saline, i.p.) for 7 days, followed by a washout period of at least 7 days. Rats were randomized so that all rats in a cage received different drug treatments.

[0075] Data are expressed as percentage of correct responses (±SEM), and values ​​for the initial and reversal phases are presented for different drug treatment groups (Figure 2). Percent correct responses data were used to determine whether drugs significantly affected response accuracy (e.g., which may reflect cognitive impairment). Statistical significance was estimated at P<0.05 and was determined using one-way analysis of variance to detect main effects of drug treatment in the initial and reversal phases. The total number of lever presses recorded during both the initial and reversal phases was not significantly different after treatment with either vehicle or Example 1 (Table 2), confirming the absence of nonspecific effects on general responding in this study. Table 2. Effects of acute treatment with Example 1 (1.0, 3.0, 10.0 & 30.0 mg / kg, po) in scPCP-treated rats (2 mg / kg, ip, twice daily for 7 days, followed by a washout period of at least 7 days) on general performance in the reversal learning task. Data are expressed as the mean total number of lever presses during the initial and reversal phases of the reversal learning task (± SEM (n = 6-9)). [Table 8] [Brief explanation of the drawings]

[0076] [Figure 1] Figure 1 shows predicted target engagement for the effective dose of Example 1 from allometric scaling across mouse, rat, dog, and monkey species after oral administration. Data are presented as a function of calculated % M1 receptor target engagement, where 50% corresponds to an unbound exposure equivalent to the recombinant human EC50 (86 nM or 28 ng / mL). Based on the measured equivalent unbound plasma:brain distribution profile (Kpuu=1), the exposures shown represent either the plasma or brain compartment. [Figure 2] Figure 2 shows the effects of acute treatment with Example 1 (1.0, 3.0, 10.0, and 30.0 mg / kg, po) on performance in a reversal learning task in scPCP-treated rats (2 mg / kg, ip, twice daily for 7 days, followed by at least a 7-day washout period). Data are presented as mean correct responding % ± SEM (n = 6-9). The dashed line separates the initial (left) and reversal (right) phases of the task. Data were analyzed by one-way ANOVA followed by LSD test. ***P < 0.001; the correct responding % in the reversal phase of the task was significantly decreased compared to the scPCP + vehicle group. #P < 0.05; ##P = 0.01; ###P < 0.001; the correct responding % in the reversal phase of the task was significantly increased compared to the scPCP + vehicle group.

[0077] equivalent The foregoing examples are given for the purpose of illustrating the present invention and should not be construed as imposing any limitation on the scope of the present invention. It will be readily apparent that many modifications and variations can be made to the specific embodiments of the invention described above and illustrated in the examples without departing from the principles underlying the invention. All such modifications and variations are intended to be embraced by this application.

Claims

1. Formula (1): 【Chemical 1】 （１） or a salt thereof.

2. Formula (2): 【Chemistry 2】 （２） The compound according to claim 1 or a salt thereof,

3. A salt of the compound according to claim 1 or 2.

4. A pharmaceutically acceptable salt of the compound of claim 1 or 2.

5. 3. An acid addition salt of the compound according to claim 1 or 2.

6. 3. The hydrochloride salt of the compound of claim 1 or 2.

7. 3. The monohydrochloride salt of the compound of claim 1 or 2.

8. Formula (2b): 【Chemistry 3】 (2b) 2. The compound of claim 1, wherein

9. Formula (2c): 【Chemistry 4】 (2c) 2. The compound of claim 1, wherein

10. 10. A pharmaceutical composition comprising a compound of claim 1, 2, 8 or 9, or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable excipient.

11. 10. A muscarinic M1 and M4 receptor agonist comprising a compound according to claim 1, 2, 8 or 9 or a pharmaceutically acceptable salt thereof.

12. 11. The pharmaceutical composition of claim 10 for use in treating a cognitive or psychiatric disorder, or for treating or reducing the severity of acute, chronic, neuropathic, or inflammatory pain.

13. 13. The pharmaceutical composition of claim 12, wherein the disorder is Alzheimer's disease.

14. 13. The pharmaceutical composition of claim 12, wherein the disorder is dementia with Lewy bodies.

15. 13. The pharmaceutical composition of claim 12, wherein the disorder is schizophrenia.

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