Pharmaceutical compounds

Abstract

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

Description

[0001] This invention relates to a new class of heterocyclic compounds, their salts, pharmaceutical compositions containing them, and their use in human treatment. In particular, this invention relates to a class of compounds that are agonists of muscarinic M1 and / or M4 receptors, and therefore can be used to treat Alzheimer's disease, schizophrenia, cognitive impairment, and other diseases mediated by muscarinic M1 / M4 receptors, including but not limited to treating or relieving pain. Background of the Invention

[0003] Muscarinic acetylcholine receptors (mAChRs) are members of the G protein-coupled receptor superfamily that mediate the effects of the neurotransmitter acetylcholine in the central and peripheral nervous systems. Five mAChR subtypes, M1 through M5, have been cloned. M1 mAChRs are primarily expressed postsynaptactically in the cortex, hippocampus, striatum, and thalamus; M2 mAChRs are mainly located in the brainstem and thalamus, although they are also found in the cortex, hippocampus, and striatum, where they are located at cholinergic synaptic terminals (Langmead et al., 2008 Br J Pharmacology). However, M2 mAChRs are also peripherally expressed in cardiac tissue (where they mediate vagal innervation of the heart) and in smooth muscle and exocrine glands. M3 mAChRs are expressed at relatively low levels in the CNS, but are widely expressed in smooth muscle and glandular tissues such as sweat glands and salivary glands (Langmead et al., 2008 Br J Pharmacology).

[0004] Muscarinic receptors in the central nervous system, particularly the M1 mAChR, play a crucial role in mediating higher cognitive processing. Diseases associated with cognitive impairment, such as Alzheimer's disease, are accompanied by the loss of cholinergic neurons in the basal forebrain (Whitehouse et al., 1982, Science). In schizophrenia, which also features cognitive impairment, mAChR density is reduced in the prefrontal cortex, hippocampus, and caudate nucleus of individuals with schizophrenia (Dean et al., 2002, Mol Psychiatry). Furthermore, in animal models, blockage or damage to central cholinergic pathways leads to severe cognitive deficits, and non-selective mAChR antagonists have been shown to induce psychotic-like 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 efficacy in treating symptomatic cognitive decline in clinical practice, but produce dose-limiting side effects due to stimulation of peripheral M2 and M3 mAChRs, including gastrointestinal motility disorders, bradycardia, nausea, and vomiting (http: / / www.drugs.com / pro / donepezil.html; http: / / www.drugs.com / pro / rivastigmine.html).

[0005] Further discoveries aimed to identify direct M1 mAChR agonists to enhance cognitive function. These efforts resulted in the identification of a range of agonists, exemplified by compounds such as xanomeline, AF267B, sabcomeline, milameline, and cevimeline. Many of these compounds have been shown to be highly effective in preclinical cognitive models in rodents and / or non-human primates. Milameline demonstrated efficacy against scopolamine-induced deficits in working and spatial memory in rodents; sabcomeline demonstrated efficacy in visual object discrimination tasks in marmosets; and xanomeline reversed cognitive deficits in a passive avoidance paradigm induced by mAChR antagonists.

[0006] Alzheimer's disease (AD) is the most common neurodegenerative disease (affecting 26.6 million people worldwide in 2006), impacting older adults and causing severe memory loss and cognitive impairment. The etiology of this disease is complex, but it is characterized by two hallmark brain sequelae: the aggregation of amyloid plaques, primarily composed of amyloid-β peptide (Aβ), and neurofibrillary tangles formed by hyperphosphorylated tau protein. Aβ accumulation is considered a major feature of AD progression; therefore, many current putative therapies for AD 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: non-amyloidogenic and amyloidogenic. Cleavage of APP by γ-secretase is common to both pathways, but in the former, APP is cleaved by α-secretase to produce soluble APPα. The cleavage site is within the Aβ sequence, thus preventing its formation. However, in the amyloidogenic pathway, APP is cleaved by β-secretase to produce soluble APPβ as well as Aβ. In vitro studies have shown that mAChR agonists can promote APP processing via the soluble, non-amyloid protease pathway. In vivo studies have shown that the mAChR agonist AF267B alters disease-like pathology in 3×TgAD transgenic mice (a model of different components of Alzheimer's disease) (Caccamo et al., 2006 Neuron). Finally, the mAChR agonist cerebrospinal fluid (Cevimeline) has been shown to slightly but significantly reduce Aβ levels in the cerebrospinal fluid of Alzheimer's patients, demonstrating potential disease-modifying efficacy (Nitsch et al., 2000 Neuron).

[0007] Furthermore, preclinical studies have shown that mAChR agonists exhibit atypical antipsychotic drug-like characteristics in a range of preclinical paradigms. The mAChR agonist phenomenoline reversed many dopamine-driven behaviors, including amphetamine-induced movement in rats, apomorphine-induced climbing in mice, dopamine agonist-driven turning in rats with unilateral 6-OH-DA lesions, and amphetamine-induced motor restlessness (without EPS tendency) in monkeys. It has also been shown that inhibition of A10, but not A9, induces dopamine cell firing and conditioned avoidance, and induces c-fos expression in the prefrontal cortex and nucleus accumbens of rats, but not in the striatum. These data all suggest atypical antipsychotic drug-like characteristics (Mirza et al., 1999 CNSDrug Rev). Muscarinic receptors are also involved in the neurobiology of addiction. The reinforcing effects of cocaine and other addictive substances are mediated by the mesolimbic dopamine system, in which behavioral and neurochemical studies have shown that cholinergic muscarinic receptor subtypes play an important role in the regulation of dopaminergic neurotransmission. For example, M(4)(- / -) mice have shown significantly enhanced reward-driven behavior due to cocaine exposure (Schmidt et al., Psychopharmacology (2011) Aug; 216(3):367-78). Furthermore, zenomeprazole has been shown to block the effects of cocaine in these models.

[0008] Muscarinic receptors are also involved in motor control and potentially represent novel treatments for movement disorders such as Parkinson's disease, ADHD, Huntington's disease, Tourette syndrome, and other syndromes associated with dopaminergic dysfunction, which is a potential driver of these diseases.

[0009] Zanomeprazole, sacram, melamram, and cevimerin have all progressed to various stages of clinical development for the treatment of Alzheimer's disease and / or schizophrenia. Phase II clinical studies with zanomeprazole have demonstrated its efficacy in various cognitive symptom domains, including behavioral disorders and hallucinations associated with Alzheimer's disease (Bodick et al., 1997 Arch Neurol). The compound has also been evaluated in a small Phase II study of schizophrenia, showing significant reductions in both positive and negative symptoms compared to a placebo-controlled trial (Shekhar et al., 2008 Am J Psych). However, in all clinical studies, zanomeprazole and other related mAChR agonists have shown unacceptable safety margins regarding cholinergic side effects, including nausea, gastrointestinal pain, diarrhea, sweating (excessive sweating), salivation (excessive salivation), syncope, and bradycardia.

[0010] Muscarinic receptors are involved in central and peripheral pain. Pain can be classified into three distinct types: acute pain, inflammatory pain, and neuropathic pain. Acute pain plays a vital protective role in keeping an organism safe from stimuli that may cause tissue damage; however, postoperative pain management is necessary. Inflammatory pain can occur due to a variety of causes, including tissue damage, autoimmune responses, and pathogen invasion, and is triggered by the action of inflammatory mediators such as neuropeptides and prostaglandins that induce neuroinflammation and pain. Neuropathic pain is associated with abnormal pain sensations in response to non-painful stimuli. Neuropathic pain is associated with a variety of different diseases / traumas, such as spinal cord injury, multiple sclerosis, diabetes (diabetic neuropathy), and viral infections (e.g., HIV or herpes). It is also common in cancer, both due to the disease itself and as a side effect of chemotherapy. Activation of muscarinic receptors has been shown to be analgesic in many pain states through activation of receptors in higher pain centers in the spinal cord and brain. Increasing endogenous levels of acetylcholine via acetylcholinesterase inhibitors, and directly activating muscarinic receptors with agonists or allosteric modulators, have been shown to have analgesic activity. Conversely, blocking muscarinic receptors with antagonists or using gene knockout mice increases pain sensitivity. Evidence regarding the role of the M1 receptor in pain was reviewed by D.F. Fiorino and M. Garcia-Guzman in 2012.

[0011] Recently, a small number of compounds have been identified that exhibit improved selectivity for the M1 mAChR isotype relative to peripherally expressed mAChR isotypes (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 selectivity relative to the M3 mAChR isotype, some of these compounds retain significant agonist activity for both this isotype and the M2 mAChR isotype. In this paper, we describe a series of compounds that unexpectedly exhibit high selectivity for M1 and / or M4 mAChRs relative to the M2 and M3 receptor isotypes. Summary of the Invention

[0012] This invention provides compounds having activity as muscarinic M1 and / or M4 receptor agonists. More specifically, this invention provides compounds exhibiting selectivity for M1 and / or M4 receptors relative to M2 and M3 receptor subtypes.

[0013] Therefore, the present invention provides a compound of formula (1) or a salt thereof:

[0014]

[0015] The present invention also provides a compound of formula (1a):

[0016] Where X represents a salt. The present invention also provides a compound of formula (1b):

[0017]

[0018] The present invention also provides compounds of formula (2):

[0019] Or its salt.

[0020] The present invention also provides a compound of formula (2a):

[0021] Where X represents salt.

[0022] The present invention also provides a compound of formula (2b):

[0023]

[0024] The present invention also provides a compound of formula (2c):

[0025]

[0026] The compound of formula (1) or formula (2) may be a pharmaceutically acceptable salt.

[0027] The compound of formula (1) or formula (2) can be an acid addition salt.

[0028] The compound of formula (1) or formula (2) can be a hydrochloride salt.

[0029] The compound of formula (1) or formula (2) can be a monohydrochloride salt.

[0030] The compound of formula (1) or formula (2) can be a monohydrochloride salt.

[0031] In compounds of formula (1a) or (2a), X may be a pharmaceutically acceptable salt.

[0032] In compounds of formula (1a) or (2a), X can be an acid addition salt.

[0033] In compounds of formula (1a) or (2a), X can be a hydrochloride salt.

[0034] In compounds of formula (1a) or (2a), X can be a monohydrochloride salt.

[0035] The compound of formula (1) or formula (2) can be a hydrochloride salt.

[0036] X can be a hydrochloride salt. X can be a monohydrochloride salt. X can be a monohydrochloride salt monohydrate.

[0037] The compound may be N-tert-butyl-1-{8-[3-(trifluoromethyl)-1,2,4-oxadiazol-5-yl]-8-azabicyclo[3.2.1]octane-3-yl}piperidine-4-carboxamide.

[0038] The compound may be N-tert-butyl-1-{(1R,3r,5S)-8-[3-(trifluoromethyl)-1,2,4-oxadiazol-5-yl]-8-azabicyclo[3.2.1]octane-3-yl}piperidine-4-carboxamide.

[0039] The compound may be a salt of N-tert-butyl-1-{8-[3-(trifluoromethyl)-1,2,4-oxadiazol-5-yl]-8-azabicyclo[3.2.1]octane-3-yl}piperidine-4-carboxamide.

[0040] The compound may 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]octane-3-yl}piperidine-4-carboxamide.

[0041] The compound may be a pharmaceutically acceptable salt of N-tert-butyl-1-{8-[3-(trifluoromethyl)-1,2,4-oxadiazol-5-yl]-8-azabicyclo[3.2.1]octane-3-yl}piperidine-4-carboxamide.

[0042] The compound may 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]octane-3-yl}piperidine-4-carboxamide.

[0043] The compound may be N-tert-butyl-1-{8-[3-(trifluoromethyl)-1,2,4-oxadiazol-5-yl]-8-azabicyclo[3.2.1]octane-3-yl}piperidine-4-carboxamide hydrochloride.

[0044] The compound may be N-tert-butyl-1-{(1R,3r,5S)-8-[3-(trifluoromethyl)-1,2,4-oxadiazol-5-yl]-8-azabicyclo[3.2.1]octane-3-yl}piperidine-4-carboxamide hydrochloride.

[0045] The compound may be N-tert-butyl-1-{8-[3-(trifluoromethyl)-1,2,4-oxadiazol-5-yl]-8-azabicyclo[3.2.1]octane-3-yl}piperidine-4-carboxamide monohydrochloride.

[0046] The compound may be N-tert-butyl-1-{(1R,3r,5S)-8-[3-(trifluoromethyl)-1,2,4-oxadiazol-5-yl]-8-azabicyclo[3.2.1]octane-3-yl}piperidine-4-carboxamide monohydrochloride.

[0047] The compound may be N-tert-butyl-1-{8-[3-(trifluoromethyl)-1,2,4-oxadiazol-5-yl]-8-azabicyclo[3.2.1]octane-3-yl}piperidine-4-carboxamide monohydrochloride monohydrate.

[0048] The compound may be N-tert-butyl-1-{(1R,3r,5S)-8-[3-(trifluoromethyl)-1,2,4-oxadiazol-5-yl]-8-azabicyclo[3.2.1]octane-3-yl}piperidine-4-carboxamide monohydrochloride monohydrate.

[0049] definition

[0050] In this application, unless otherwise stated, the following definitions apply.

[0051] The term "treatment" in connection with the use of compounds of formula (1), (1a), (1b), (2), (2a), (2b), or (2c) is used to describe any form of intervention in which a compound is administered to an individual who has, is at risk of having, or is potentially at risk of having, the disease or condition in question. Therefore, the term "treatment" encompasses both prophylactic treatment and treatment in which measurable or detectable symptoms of a disease or condition are manifested.

[0052] As used herein, the term "effective therapeutic amount" (e.g., in relation to a treatment of a disease or condition) refers to the amount of a compound that effectively produces the desired therapeutic effect. For example, if the condition is pain, the effective therapeutic amount is an amount sufficient to provide the desired level of pain relief. The desired level of pain relief could be, for example, complete elimination of pain or reduction of the severity of pain.

[0053] Salts

[0054] The compounds described herein may exist in the form of salts, such as acid addition salts, or in some cases, salts of organic and inorganic bases, such as carboxylates, sulfonates, and phosphates. All such salts are within the scope of this invention, and references to compounds of formulas (1) and (2) include salt forms of compounds as defined herein.

[0055] Salts are usually salts formed by the addition of acids.

[0056] The salts of the present invention can be synthesized from parent compounds containing a basic or acidic moiety by conventional chemical methods, such as those described in Pharmaceutical Salts: Properties, Selection, and Use, P. Heinrich Stahl (ed.), Camille G. Wermuth (ed.), ISBN: 3-90639-026-8, Hardcover, 388 pages, August 2002. Typically, such salts can be prepared by reacting the free acidic or basic form of these compounds with a suitable base or acid in water, in an organic solvent, or in a mixture of both; typically, non-aqueous media such as diethyl ether, ethyl acetate, ethanol, isopropanol, or acetonitrile are used.

[0057] Acid addition salts can be formed from a variety of acids (inorganic and organic). Examples of acid addition salts falling within the scope of this invention include mono- or di-salts formed with an acid selected from 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-acetaminobenzoic acid, butyric acid, (+)camphoric acid, camphorsulfonic acid, (+)-(1S)-camphor-10-sulfonic acid, decanoic acid, hexanoic acid, octanoic acid, cinnamic acid, citric acid, cyclohexanesulfonic acid, dodecyl sulfate, ethane-1,2-disulfonic acid, ethanesulfonic acid, 2-hydroxyethanesulfonic acid, formic acid, fumaric acid, galactopyric acid, gentian acid, glucoheponic acid, D-gluconic acid, glucuronic acid (e.g., D-glucuronic acid), glutamic acid (e.g., L-glutamic acid), α-ketoglutarate. Acids, glycolic acid, hippuric acid, hydrohalic acids (e.g., hydrobromic acid, hydrochloric acid, hydroiodic acid), hydroxyethanesulfonic 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, dihydroxynaphthoic 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, thiocyanate, p-toluenesulfonic acid, undecenoic acid and valeric acid, as well as acylated amino acids and cation exchange resins.

[0058] The amine functional groups in the compounds described herein can form quaternary ammonium salts, for example, by reacting with an alkylating agent according to methods well known to those skilled in the art. Such quaternary ammonium compounds are within the scope of this invention.

[0059] The compounds of the present invention can exist as a single salt or a disalt, depending on the pKa of the acid that forms the salt.

[0060] The salt forms of the compounds of this invention are generally 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, non-pharmaceutically acceptable salts can also be prepared in intermediate forms and then converted into pharmaceutically acceptable salts. Such non-pharmaceutically acceptable salt forms (which can be used, for example, for the purification or isolation of the compounds of this invention) also form part of this invention.

[0061] Stereoisomers

[0062] References to compounds of formulas (1), (1a) and (1b) include all possible stereoisomers (e.g., enantiomers, epiomers and diastereomers, including in- and out-isomers), either as a single isomer, or as a mixture (e.g., a racemic mixture) or two or more isomers, unless the context requires otherwise.

[0063] Therefore, the present invention provides a compound of formula (1) containing a chiral center.

[0064] Isomers can be characterized by their absolute stereochemistry using the “R and S” nomenclature developed by Cahn, Ingold, and Prelog, see Advanced Organic Chemistry, 4th ed., John Wiley & Sons, New York, 1992, pp. 109-114; also see Cahn, Ingold & Prelog, Angew. Chem. Int. Ed. Engl., 1966, 5, 385-415. Stereoisomers can be separated by a variety of techniques, including chiral chromatography (chromatography on chiral supports), 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-toluyl-L-tartaric acid, (+)-mandelic acid, (-)-malic acid, and (-)-camphorsulfonic acid, separating the diastereomeric isomers by preferential crystallization, and then dissociating the salts to obtain the individual enantiomers of the free base.

[0065] When the compounds of the present invention exist in two or more stereoisomers, one of the diastereomers may exhibit advantages over the other, for example, in terms of biological activity. Therefore, in some cases, it may be desirable to use only one of the multiple diastereomers as a therapeutic agent.

[0066] Therefore, the present invention provides compositions comprising compounds 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).

[0067] In one general embodiment, 99% or more (e.g., substantially all) of the total amount of the compound (or the compound used) is present as a single isomer.

[0068] For example, in one embodiment, the compound exists as a single diastereomer and the compound has a plane of symmetry.

[0069] Isotopes

[0070] The compounds of this invention may contain one or more isotopic substitutions, and references to a particular element include all isotopes of that element within their scope. For example, references to hydrogen include... 1 H, 2 H(D) and 3 H(T). Similarly, references to carbon and oxygen, within their respective scopes, include... 12 C 13 C and 14 C and 16 O and 18 O.

[0071] In a similar manner, references to a particular functional group also include isotopic variations within their scope, unless the context otherwise requires. For example, references to alkyl groups such as tert-butyl also cover variations in which one or more hydrogen atoms in the group are in the form of deuterium or tritium isotopes, such as in tert-butyl (all-deuterium-tert-butyl) in which all nine hydrogen atoms are in the form of deuterium isotopes.

[0072] The isotope can be radioactive or non-radioactive. The compound may not contain a radioactive isotope. Such a compound is preferably intended for therapeutic use. However, the compound may contain one or more radioactive isotopes. Compounds containing such radioactive isotopes can be used in diagnostic settings.

[0073] solvates

[0074] 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., crystal structure) of the compounds of the present invention. Examples of such solvents include water, alcohols (such as ethanol, isopropanol, and butanol), and dimethyl sulfoxide. Solvates can be prepared by recrystallizing the compounds of the present invention with a solvent or a mixture of solvents containing a solvating solvent. Whether a solvate has been formed in any given case can be determined by analyzing the crystals of the compound using well-known standard techniques such as thermogravimetric analysis (TGE), differential scanning calorimetry (DSC), and X-ray crystallography. Solvates can be stoichiometric or non-stoichiometric. Particularly preferred solvates are hydrates, and examples of hydrates include hemihydrates, monohydrates, and dihydrates.

[0075] Therefore, the present invention provides:

[0076] Compounds in solvate form.

[0077] The solvate is a hydrate compound.

[0078] The solvate is a hydrated compound.

[0079] 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, 2nd ed., published by SSCI, Inc. of West Lafayette, IN, USA, 1999, ISBN 0-967-06710-3.

[0080] Alternatively, the compounds of the present invention may be anhydrous, rather than existing as hydrates. Therefore, the present invention provides the compounds of the present invention in anhydrous form (e.g., anhydrous crystalline form).

[0081] Crystalline and amorphous forms

[0082] The compound can exist in a crystalline or amorphous (e.g., amorphous) state. Whether a compound exists in a crystalline state can be readily determined using standard techniques such as X-ray powder diffraction (XRPD). Crystals and their crystal structures can be characterized using a variety of techniques, including single-crystal X-ray crystallography, X-ray powder diffraction (XRPD), differential scanning calorimetry (DSC), and infrared spectroscopy, such as Fourier transform infrared spectroscopy (FTIR). The behavior of crystals under different humidity conditions can be analyzed using gravimetric adsorption studies and XRPD. The determination of the crystal structure of a compound can be performed 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 of single crystals. In amorphous solids, the three-dimensional structure that is usually present in crystalline forms does not exist, and in amorphous forms, the positions of molecules relative to each other are essentially random, see, for example, Hancock et al., J. Pharm.sci. (1997), 86, 1.

[0083] Therefore, the present invention provides:

[0084] A compound in crystalline form.

[0085] A compound, wherein the compound is:

[0086] (a) 50% to 100% crystallization, and more particularly at least 50% crystallization, or at least 60% crystallization, or at least 70% crystallization, or at least 80% crystallization, or at least 90% crystallization, or at least 95% crystallization, or at least 98% crystallization, or at least 99% crystallization, or at least 99.5% crystallization, or at least 99.9% crystallization, such as 100% crystallization.

[0087] A compound in an amorphous form.

[0088] Complexes and inclusion compounds

[0089] It also includes complexes of the compounds of the present invention (e.g., inclusion complexes or inclusion compounds with compounds such as cyclodextrin, or complexes with metals).

[0090] Therefore, the present invention provides compounds in the form of complexes or inclusion compounds.

[0091] Bioactivity and therapeutic uses

[0092] The compounds of the present invention exhibit activity as muscarinic M1 and M4 receptor agonists. The muscarinic activity of the compounds can be determined using the phosphate-ERK1 / 2 assay described in Example A below.

[0093] A significant advantage of the compounds of the present invention is their high selectivity for M1 and M4 receptors relative to M2 and M3 receptor subtypes. The compounds of the present invention are not agonists of the M2 and M3 receptor subtypes. For example, although the compounds of the present invention typically have a pEC of at least 6 (preferably at least 6.5) for the M1 receptor in the functional assays described in Example A. 50 Values ​​and E values ​​greater than 80 (preferably greater than 90) max However, when tested for the M2 and M3 subtypes in the functional assays of Example A, they may have a pEC value of less than 5. 50 Values ​​and less than 20% of E max value.

[0094] Regarding the compounds of the present invention, the present invention also provides:

[0095] A compound used in pharmaceuticals.

[0096] A compound used as a muscarinic M1 and / or M4 receptor agonist.

[0097] A compound, which is a muscarinic M1 receptor agonist, has a pEC greater than 6.9 for the M1 receptor in the assays of Example A herein or substantially similar assays. 50 and at least 80% of E max .

[0098] A compound, which is pEC 50 Muscarinic M1 receptor agonists with a strength greater than 7.0.

[0099] A compound having an E1 affinity of at least 90 for the M1 receptor. max .

[0100] A compound that is a muscarinic M1 and M4 receptor agonist, exhibiting a pEC greater than 6.0 for the M4 receptor in the assays performed in Example A herein or in substantially similar assays. 50 .

[0101] A compound that is selective for M1 and M4 receptors compared to muscarinic M2 and M3 receptors.

[0102] A compound having a pEC value of less than 5 for muscarinic M2 and M3 receptor subtypes. 50 and E less than 30 max .

[0103] A compound for treating diseases or conditions mediated by muscarinic M1 and / or M4 receptors.

[0104] Due to their muscarinic M1 and M4 receptor agonist activity, the compounds of the present invention can be used to treat Alzheimer's disease, Lewy body dementia, schizophrenia and other psychotic disorders, cognitive impairment and other diseases mediated by muscarinic M1 and / or M4 receptors, and can also be used to treat various types of pain.

[0105] Therefore, regarding the compounds of the present invention, the present invention also provides:

[0106] A compound used to treat cognitive impairment or psychotic disorders.

[0107] A compound for treating cognitive impairment or psychotic disorders, wherein the cognitive impairment or psychotic disorder includes, arises from, or is associated with conditions selected from: cognitive impairment, mild cognitive impairment (MCI), (including amnestic and non-amnestic MCI, and including mild cognitive impairment due to Alzheimer's disease and / or prodromal Alzheimer's disease), frontotemporal dementia, vascular dementia, Lewy body dementia, early-onset Alzheimer's disease, senile dementia, Friedlich ataxia, Down syndrome, Huntington's disease, hyperkinesis, mania, Tourette syndrome, Alzheimer's disease (including prodromal Alzheimer's disease and those caused by the U.S. Food and Drug Administration). Early Alzheimer's disease, as defined by the FDA's "Early Alzheimer's Disease: Developed Drugs for Treatment," comprises stages 1, 2, and 3 (available at fda.gov / downloads / Drugs / GuidanceComplianceRegulatoryInformation / Guidances / UCM596728.pdf), progressive supranuclear palsy, and impairment of cognitive function, including attention, orientation, learning disabilities, memory (i.e., memory impairment, amnesia, forgetfulness disorder, transient amnesia syndrome, and age-related memory impairment), and language function. Cognitive impairment caused by stroke, Huntington's disease, Pick's disease, AIDS-related dementia or other dementia states such as multi-infarct dementia, alcoholic dementia, hypothyroid dementia, and dementia associated with other degenerative diseases such as cerebellar atrophy and amyotrophic lateral sclerosis; other acute or subacute conditions that can cause cognitive decline, such as delirium or depression (pseudodementia), trauma, head trauma, age-related cognitive decline, stroke, neurodegeneration, drug-induced states, neurotoxins, age-related cognitive impairment, autism-related cognitive impairment, Down syndrome, cognitive deficits associated with psychosis, and post-electroconvulsive therapy. Cognitive impairment related to substance abuse or withdrawal, including nicotine, cannabis, amphetamines, cocaine; attention deficit hyperactivity disorder (ADHD) and movement disorders, such as Parkinson's disease, psychotic-induced Parkinsonian syndrome and tardive dyskinesia; schizophrenia, schizophrenia-like disorders, psychotic depression, mania, acute mania, paranoia, hallucinations and delusions, personality disorders, obsessive-compulsive disorder, schizotypal personality disorder, delusions, psychosis due to malignancy, metabolic disorders, endocrine disorders or narcolepsy, psychosis due to substance abuse or withdrawal, bipolar disorder and schizoaffective disorder.

[0108] A compound used to treat Alzheimer's disease.

[0109] A compound used to treat Lewy body dementia.

[0110] A compound used to treat schizophrenia.

[0111] A method for treating cognitive impairment in an individual (e.g., a mammalian patient, such as a human, such as a person requiring such treatment), the method comprising administering a therapeutically effective dose of the compound of the present invention.

[0112] A method for treating cognitive impairment in an individual (e.g., a mammalian patient, such as a human, such as a person requiring such treatment), the method comprising administering a therapeutically effective dose of the compound of the present invention, wherein the cognitive impairment includes a condition as defined above, is caused by a condition as defined above, or is related to a condition as defined above.

[0113] A method for treating cognitive impairment in an individual (e.g., a mammalian patient, such as a human, such as a person requiring such treatment), the method comprising administering a therapeutically effective dose of the compound of the present invention, wherein the cognitive impairment is caused by or related to Alzheimer's disease.

[0114] A method for treating cognitive impairment in an individual (e.g., a mammalian patient, such as a human, such as a person requiring such treatment), the method comprising administering a therapeutically effective dose of the compound of the present invention, wherein the cognitive impairment is Lewy body dementia.

[0115] A method for treating cognitive impairment in an individual (e.g., a mammalian patient, such as a human, such as a person requiring such treatment), the method comprising administering a therapeutically effective dose of the compound of the present invention, wherein the cognitive impairment is schizophrenia.

[0116] The use of the compounds of the present invention in the preparation of medicaments for the treatment of cognitive impairment.

[0117] The use of the compounds of the present invention in the preparation of medicaments for treating cognitive impairment, wherein the cognitive impairment includes conditions as defined above, caused by conditions as defined above, or related to conditions as defined above.

[0118] The use of the compounds of the present invention in the preparation of medicaments for treating cognitive impairment, wherein the cognitive impairment includes Alzheimer's disease, caused by Alzheimer's disease, or related to Alzheimer's disease.

[0119] The use of the compounds of the present invention in the preparation of medicaments for treating cognitive impairment, wherein the cognitive impairment includes Lewy body dementia, caused by Lewy body dementia, or associated with Lewy body dementia.

[0120] The use of the compounds of the present invention in the preparation of medicaments for treating cognitive impairment, wherein the cognitive impairment includes schizophrenia, schizophrenia-induced schizophrenia, or schizophrenia-related schizophrenia.

[0121] Compounds used to treat acute, chronic, neuropathic or inflammatory pain, arthritis, migraine, cluster headache, trigeminal neuralgia, herpetic neuralgia, general neuralgia, visceral pain, osteoarthritis pain, postherpetic neuralgia, diabetic neuropathy, radicular pain, sciatica, back pain, head or neck pain, severe or intractable pain, nociceptive pain, breakthrough pain, postoperative pain, or cancer pain, or to reduce their severity.

[0122] 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, osteoarthritis pain, postherpetic neuralgia, diabetic neuropathy, radicular pain, sciatica, back pain, head or neck pain, severe or intractable pain, nociceptive pain, breakthrough pain, postoperative pain, or cancer pain, said method comprising administering a therapeutically effective dose of the compound of the present invention.

[0123] A compound used to treat peripheral conditions, such as lowering intraocular pressure in glaucoma and treating dry eye and dry mouth, including Sjogren's Syndrome.

[0124] A method for treating peripheral conditions, such as reducing intraocular pressure in glaucoma and treating dry eye and dry mouth, including Sjögren's syndrome, said method comprising administering a therapeutically effective dose of the compound of the present invention.

[0125] The use of the compounds of the present invention in the preparation of medicaments for treating or reducing the severity of the following conditions: acute, chronic, neuropathic or inflammatory pain, arthritis, migraine, cluster headache, trigeminal neuralgia, herpetic neuralgia, generalized neuralgia, visceral pain, osteoarthritis pain, postherpetic neuralgia, diabetic neuropathy, radicular pain, sciatica, back pain, head or neck pain, severe or intractable pain, nociceptive pain, breakthrough pain, postoperative pain or cancer pain, or in the preparation of medicaments for treating peripheral conditions, such as lowering intraocular pressure in glaucoma and treating dry eye and dry mouth, including Sjögren's syndrome.

[0126] The compounds of this invention are used to treat skin lesions, such as skin lesions caused by pemphigus vulgaris, herpetic dermatitis, bullous pemphigoid, and other blistering skin conditions.

[0127] The compounds of this invention are intended for the treatment, prevention, improvement or reversal of conditions related to gastrointestinal function and motility, such as functional dyspepsia, irritable bowel syndrome, gastroesophageal acid reflux (GER) and esophageal motility disorders, gastroparesis symptoms and chronic diarrhea.

[0128] The compounds of the present invention are intended for use in the treatment of olfactory dysfunction, such as Bosma-Henkin-Christiansen syndrome, chemical poisoning (e.g., selenium and silver), hypopituitarism, Kallmann syndrome, skull fractures, tumor treatment, and hypoactive thyroid glands.

[0129] The use of the compounds of this invention for the treatment of addiction.

[0130] The compounds of this invention are intended for the treatment of movement disorders such as Parkinson's disease, ADHD, Huntington's disease, Tourette syndrome, and other syndromes associated with dopaminergic dysfunction as a potential pathogenic factor driving these diseases.

[0131] The compounds of this invention are intended for the treatment of behavioral and psychological symptoms of dementia (BPSD; including agitation, verbal aggression, physical aggression, depression, anxiety, abnormal motor behavior, elevated mood, irritability, emotional blunting, disinhibition, impulsivity, delusions, hallucinations, sleep changes, and appetite changes).

[0132] The compounds of the present invention include those shown in Examples 1, 1-1 and 1-2 below.

[0133]

[0134] Method for preparing the compounds of the present invention

[0135] The compounds of the present invention can be prepared according to synthetic methods known to those skilled in the art and described herein.

[0136] Methods for preparing compounds as defined above are also provided, which may include any one of A, B, or C:

[0137] (A) Compound of formula (10):

[0138]

[0139] Reaction with compound of formula (11) under reducing amination conditions:

[0140]

[0141] or

[0142] (B) Compound of formula (12):

[0143]

[0144] Reaction with amines of formula (CH3)3CNH2, where R represents a suitable group, such as methyl or ethyl; or

[0145] (C) Compound of formula (13):

[0146]

[0147] It reacts with amines of formula (CH3)3CNH2.

[0148] This method is well known to those skilled in the art. Examples of synthetic procedures for converting one functional group into another are listed 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's Reagents for Organic Synthesis, Volumes 1-17, John Wiley, edited by Mary Fieser (ISBN: 0-471-58283-2).

[0149] In the reactions described above, it may be necessary to protect one or more groups to prevent the reaction from occurring at undesirable positions on the molecule. Examples of protecting groups, as well as methods for protecting and deprotecting functional groups, can be found in Greene's Protective Groups in Organic Synthesis, 5th Edition, edited by Peter GMWuts and John Wiley, 2014 (ISBN: 9781118057483).

[0150] Compounds prepared by the above method can be separated and purified by any of a variety of methods known to those skilled in the art, and examples of such methods include recrystallization and chromatographic techniques, such as column chromatography (e.g., rapid chromatography), HPLC and SFC.

[0151] pharmaceutical preparations

[0152] Although the active compound can be administered alone, it is preferred to present it as a pharmaceutical composition (e.g., a formulation).

[0153] Therefore, a pharmaceutical composition is provided comprising at least one compound of the present invention as defined above and at least one pharmaceutically acceptable excipient.

[0154] The composition may be a tablet composition.

[0155] The composition may be a capsule composition.

[0156] Pharmaceutically acceptable excipients may be selected from, for example, carriers (e.g., solid, liquid, or semi-solid carriers), excipients, diluents (e.g., solid diluents, such as fillers or compatibilizers; and liquid diluents, such as solvents and cosolvents), granulators, binders, flow aids, coating agents, controlled-release agents (e.g., sustained-release or delayed-release polymers or waxes), adhesives, disintegrants, buffers, lubricants, preservatives, antifungal and antibacterial agents, antioxidants, buffers, tension modifiers, thickeners, flavoring agents, sweeteners, colorants, plasticizers, taste masking agents, stabilizers, or any other excipients conventionally used in pharmaceutical compositions.

[0157] As used herein, "pharmaceutically acceptable" means a compound, material, composition, and / or dosage form that, to a reasonable extent of medical judgment, is suitable for contact with the tissues of an individual (e.g., a human individual) without excessive toxicity, irritation, allergic reactions, or other problems or complications, and is proportionate to a reasonable benefit / risk ratio. Each excipient must also be "acceptable" in the sense of compatibility with other components of the formulation.

[0158] 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.

[0159] The pharmaceutical composition may be in any form suitable for oral, parenteral, topical, intranasal, intrabronchial, sublingual, ophthalmic, optic, rectal, vaginal, or transdermal administration.

[0160] Suitable dosage forms for oral administration include tablets (coated or uncoated), capsules (hard or soft shell), capsules, pills, lozenges, syrups, solutions, powders, granules, elixirs and suspensions, sublingual tablets, films, or patches, such as oral patches.

[0161] Tablet compositions may contain a unit dose of an active compound and an inert diluent or carrier, such as sugars or sugar alcohols; for example, lactose, sucrose, sorbitol, or mannitol; and / or non-sugar-derived diluents, such as sodium carbonate, calcium phosphate, calcium carbonate, or cellulose or its derivatives, such as microcrystalline cellulose (MCC), methylcellulose, ethylcellulose, hydroxypropyl methylcellulose, and starch, such as corn starch. Tablets may also contain standard ingredients such as binders and granulators, such as polyvinylpyrrolidone, disintegrants (e.g., expandable cross-linked polymers, such as cross-linked carboxymethyl cellulose), lubricants (e.g., stearates), preservatives (e.g., parabens), antioxidants (e.g., BHT), buffers (e.g., phosphate or citrate buffers), and effervescent agents, such as citrate / bicarbonate mixtures. Such excipients are well-known and do not require detailed discussion here.

[0162] Tablets can be designed to release the drug upon contact with gastric juices (immediate-release tablets), release the drug in a controlled manner over a prolonged period of time (controlled-release tablets), or release the drug upon contact with a specific area of ​​the gastrointestinal tract.

[0163] Pharmaceutical compositions typically comprise from about 1% (w / w) to about 95% (w / w), preferably 99% (w / w), of an active ingredient and from 5% (w / w) of a pharmaceutically acceptable excipient (e.g., as defined above) or a combination of such excipients. Preferably, the composition comprises from about 20% (w / w) to about 90% (w / w) of an active ingredient and from 80% (w / w) to 10% of a pharmaceutical excipient or a combination of excipients. Pharmaceutical compositions according to the invention may be, for example, in unit dosage forms such as ampoules, vials, suppositories, drug-loaded syringes, sugar-coated pills, powders, tablets, or capsules.

[0164] Tablets and capsules may contain, for example, 0-20% disintegrants, 0-5% lubricants, 0-5% flow aids, and / or 0-99% (w / w) fillers and / or compatibilizers (depending on the drug dosage). They may also contain 0-10% (w / w) polymer binders, 0-5% (w / w) antioxidants, and 0-5% (w / w) colorants. Furthermore, sustained-release tablets typically contain 0-99% (w / w) of controlled-release (e.g., delayed) polymers (depending on the dosage). Film coatings for tablets or capsules typically contain 0-10% (w / w) polymers, 0-3% (w / w) colorants, and / or 0-2% (w / w) plasticizers.

[0165] Parenteral preparations typically contain 0-20% (w / w) buffer, 0-50% (w / w) co-solvent, and / or 0-99% (w / w) water for injection (WFI) (depending on the dosage and whether it is lyophilized). Preparations for intramuscular reservoirs may also contain 0-99% (w / w) oil.

[0166] The drug formulation can be presented to the patient in the form of a "patient kit," which contains the entire course of treatment in a single package (usually a blister pack).

[0167] The compounds of the present invention are typically presented in unit dosage forms and therefore typically contain sufficient amounts of the compound to provide the desired level of biological activity. For example, a formulation may contain 1 nanogram to 2 grams of active ingredient, such as 1 nanogram to 2 milligrams of active ingredient. Within these ranges, specific subranges of the compounds are 0.1 milligrams to 2 grams of active ingredient (more typically 10 milligrams to 1 gram, such as 50 milligrams to 500 milligrams), or 1 microgram to 20 milligrams (such as 1 microgram to 10 milligrams, such as 0.1 milligrams to 2 milligrams of active ingredient).

[0168] For oral compositions, the unit dosage form may contain 1 mg to 2 g, more typically 10 mg to 1 g, such as 50 mg to 1 g, such as 100 mg to 1 g of active compound.

[0169] The active compound is administered to the patient in need (e.g., human or animal patient) in an amount sufficient to achieve the desired therapeutic effect (effective dose). The precise amount of compound administered can be determined by the attending physician according to standard procedures.

[0170] General Program

[0171] Without specifying the preparation route, the relevant intermediates are commercially available. Commercially available reagents can be used without further purification. Room temperature (rt) refers to approximately 22-30°C. Recordings were taken at 400 MHz on a Bruker instrument. 1 1H NMR spectroscopy. 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 peak, d = doublet, t = triplet, q = quartet, quint = quintet, td = triplet of doublet, tt = triplet of triplet, qd = quartet of doublet, ddd = doublet of doublet, ddt = doublet of doublet, m = multiplet. Coupling constants are listed as J values, measured in Hz. NMR and mass spectrometry results are corrected to account for background peaks.

[0172] LCMS Analysis

[0173] LCMS analysis of compounds under electrospray conditions was performed using the instruments and methods given in the table below:

[0174]

[0175] Preparative HPLC purification

[0176] Preparative HPLC purification was performed using a Shimadzu LC-20AP binary system with an SPD-20A UV detector. Purification techniques: [phase (column description, column length × inner diameter, particle size), solvent flow rate, gradient - % of mobile phase B in mobile phase A (over time), mobile phase (A), mobile phase (B) given].

[0177] Preparative HPLC Method A

[0178] Preparative HPLC: [reversed phase (X-BRIDGE C-18, 250×50mm, 5μm), 85mL / min, gradient 35%-70% (after 26min), 100% (after 2min), 100%-35% (after 6min), mobile phase (A): 5mM ammonium bicarbonate aqueous solution + 0.1% ammonia aqueous solution, (B): 100% acetonitrile].

[0179] Preparative HPLC Method B

[0180] Preparative HPLC: [reversed phase (X-BRIDGE C-18, 250×50mm, 5μm), 85mL / min, gradient 40%-60% (after 26min), 60% (after 4min), 100% (after 2min), 100%-40% (after 7min), mobile phase (A): 5mM ammonium bicarbonate aqueous solution + 0.1% ammonia aqueous solution, (B): 100% acetonitrile].

[0181] Abbreviations

[0182] AcOH = Acetic acid

[0183] Bn = benzyl

[0184] t-BuOH = tert-butanol

[0185] CPM = Cyclopentyl methyl ether

[0186] DCM = dichloromethane

[0187] DIPEA = N,N-Diisopropylethylamine

[0188] DMF = dimethylformamide

[0189] DMSO = dimethyl sulfoxide

[0190] ESI = Electrospray Ionization

[0191] EtOH = ethanol

[0192] h = hours

[0193] HATU = Azabenzotriazole tetramethylurea hexafluorophosphate

[0194] HPLC = High Performance Liquid Chromatography

[0195] IPA = Propyl-2-ol

[0196] LCMS = Liquid Chromatography-Mass Spectrometry

[0197] MeOH = methanol

[0198] min = minutes

[0199] 2-MTHF=2-Methyltetrahydrofuran

[0200] nm = nanometer (s)

[0201] NMO = 4-methylmorpholine 4-oxide

[0202] NMR = Nuclear Magnetic Resonance

[0203] rt = room temperature

[0204] RT = Retention Time

[0205] TEA = Triethylamine

[0206] TFA = Trifluoroacetic acid

[0207] TFAA = Trifluoroacetic anhydride

[0208] THF = Tetrahydrofuran

[0209] The prefixes n-, s-, i-, t-, and tert- have their usual meanings: positive, secondary, different, and paternal.

[0210] N-(tert-butyl)-1-((1R,3r,5S)-8-(3-(trifluoromethyl)-1,2,4-oxadiazol-5-yl)-8-azadi Synthesis of Cyclo[3.2.1]octane-3-yl)piperidine-4-carboxamide hydrochloride (Example 1-1)

[0211]

[0212]

[0213] Step 1: Synthesis of cyclopent-3-ene-1-carboxylic acid benzyl ester (intermediate 2)

[0214] To a mixture of cyclopentyl-3-en-1-carboxylic acid (CAS: 7686-77-3, intermediate 1) (25.0 g, 223 mmol) and K₂CO₃ (61.5 g, 446 mmol) in acetone (375 mL), (bromomethyl)benzene (CAS: 100-39-0) (29.1 mL, 245 mmol) was added dropwise. The mixture 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 cyclopentyl-3-en-1-carboxylate (intermediate 2) (40.1 g, 89.0%).

[0215] LCMS (System 1, Method A): (ESI)m / z 203[M+H] + RT 5.10min, 254nm.

[0216]

[0217] Step 2: Synthesis of 3,4-dihydroxycyclopentane-1-carboxylic acid benzyl ester (intermediate 3)

[0218] A mixture of cyclopentyl-3-ene-1-carboxylate (intermediate 2) (43.8 g, 217 mmol), OsO4 (2% in t-BuOH, 12.0 mL, 0.94 mmol), and 4-methylmorpholine 4-oxide (30.44 g, 260 mmol) in acetone (431 mL) was stirred at room temperature for 16 hours. The mixture was treated with a saturated aqueous solution of Na2SO3 (500 mL) and then extracted with DCM (3 × 400 mL). The combined organic layers were dried (Na2SO4) and concentrated under reduced pressure. The residue was purified by column chromatography [normal phase (silica), 0–50% (ethyl acetate in hexane)] to give 3,4-dihydroxycyclopentane-1-carboxylate (intermediate 3) (30.4 g, 59.4%).

[0219] LCMS (System 2, Method B): (ESI)m / z 237[M+H] + RT 2.38min, 224nm.

[0220]

[0221] Step 3: Synthesis of benzyl 4-oxo-2-(2-oxoethyl)butyrate (intermediate 4)

[0222] A mixture of 3,4-dihydroxycyclopentane-1-carboxylate (intermediate 3) (36.0 g, 153 mmol) and NaIO4 (48.7 g, 229 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 the mixture was then extracted with DCM (3 × 500 mL). The combined organic layers were dried (Na2SO4) and concentrated under reduced pressure to give benzyl 4-oxo-2-(2-oxoethyl)butyrate (intermediate 4) (35.8 g, 100.0%). The crude product was used without further purification.

[0223] LCMS (System 2, Method B): (ESI)m / z 233[MH] - RT 2.33min and 2.74min, 202nm.

[0224]

[0225] Step 4: (1R,3r,5S)-3-(4-((benzyloxy)carbonyl)piperidin-1-yl)-8-azabicyclo[3.2.1]octane- Synthesis of tert-butyl 8-carboxylate (intermediate 6).

[0226] A solution of (1R,3r,5S)-3-amino-8-azabicyclo[3.2.1]octane-8-carboxylic acid tert-butyl ester (CAS: 207405-68-3, intermediate 5) (34.6 g, 153 mmol) and 4-oxo-2-(2-oxoethyl)butyrate benzyl ester (intermediate 4) (35.8 g, 153 mmol) in EtOH (1400 mL) was stirred at room temperature for 30 min. Then, NaBH3CN (9.64 g, 153 mmol) and AcOH (3.0 mL, 52.5 mmol) were added, 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 (Na2SO4) and concentrated under reduced pressure. The residue was purified by column chromatography [normal phase (silica), 0-30% (ethyl acetate in hexane)] to give (1R,3r,5S)-3-(4-((benzyloxy)carbonyl)piperidin-1-yl)-8-azabicyclo[3.2.1]octane-8-carboxylic acid tert-butyl ester (intermediate 6) (41.0 g, 62.6%).

[0227] LCMS (System 2, Method B): (ESI)m / z 429[M+H] + RT 4.28min, 202nm.

[0228]

[0229] Step 5: 1-((1R,3r,5S)-8-azabicyclo[3.2.1]octane-3-yl)piperidine-4-carboxylic acid benzyl ester dihydrochloride Synthesis of salt (intermediate 7)

[0230] A solution of 1,4-dioxane (4M, 410 mL, 10 vol.) in HCl was added dropwise to a solution of (1R,3r,5S)-3-(4-((benzyloxy)carbonyl)piperidin-1-yl)-8-azabicyclo[3.2.1]octan-8-carboxylic acid tert-butyl ester (intermediate 6) (41.0 g, 96.0 mmol) in 1,4-dioxane (82.0 mL). The reaction mixture was stirred at room temperature for 2 h and then concentrated under reduced pressure. The resulting residue was azeotropically reacted with hexane (2 × 50 mL) and then ground with hexane (2 × 50 mL) to give 1-((1R,3r,5S)-8-azabicyclo[3.2.1]octan-3-yl)piperidin-4-carboxylic acid benzyl ester dihydrochloride (intermediate 7) (38.4 g, 100.0%).

[0231] LCMS (System 1, Method B): (ESI)m / z 329[M+H] + RT 3.10min, 202nm.

[0232]

[0233] Step 6: 1-((1R,3r,5S)-8-cyano-8-azabicyclo[3.2.1]octane-3-yl)piperidine-4-carboxylic acid benzyl ester Synthesis of (Intermediate 8)

[0234] Triethylamine (39.8 mL, 287 mmol) was added dropwise to a solution of 1-((1R,3r,5S)-8-azabicyclo[3.2.1]octan-3-yl)piperidin-4-carboxylate dihydrochloride (intermediate 7) (38.4 g, 96.0 mmol) in DCM (314 mL) at -20 °C, while maintaining the internal temperature in the range of -20 °C to 0 °C. The reaction was then stirred at this temperature for 30 min. A solution of cyanogen bromide (15.1 g, 144 mmol) in DCM (31.0 mL) was then added dropwise, while maintaining the internal temperature in the range of -20 °C to 0 °C. The mixture was then heated to room temperature and stirred for 16 h. The reaction mixture was diluted with a saturated aqueous solution of NaHCO3 (700 mL) and extracted with DCM (3 × 300 mL). The combined organic layers were dried (Na2SO4) and concentrated under reduced pressure. The crude product was purified by column chromatography [normal phase (neutral Al₂O₃), 0-30% (ethyl acetate in hexane)] to give 1-((1R,3r,5S)-8-cyano-8-azabicyclo[3.2.1]octane-3-yl)piperidin-4-carboxylic acid benzyl ester (intermediate 8) (16.0 g, 47.4%). LCMS (System 2, Method B): (ESI) m / z 354 [M+H] + RT 3.69min, 202nm.

[0235]

[0236] Step 7: 1-((1R,3r,5S)-8-((((tert-butoxycarbonyl)amino)oxy)(imino)methyl)-8-aza Synthesis of benzyl octane-3-yl)piperidine-4-carboxylate (intermediate 10)

[0237] At 0 °C, tert-butyl hydroxycarbamate (CAS: 36016-38-3, intermediate 9) (6.63 g, 49.8 mmol) was added to a THF (160 mL) solution of 1-((1R,3r,5S)-8-cyano-8-azabicyclo[3.2.1]octan-3-yl)piperidin-4-carboxylate (intermediate 8) (16.0 g, 45.3 mmol). The reaction mixture was stirred at 0 °C for 20 min. Then, a 2-MTHF solution of zinc chloride (1.9 M, 47.7 mL, 90.6 mmol) was slowly added 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 (Na2SO4) and concentrated under reduced pressure. The crude product was ground with hexane (2 × 50 mL) to give 1-((1R,3r,5S)-8-(((tert-butoxycarbonyl)amino)oxy)(imino)methyl)-8-azabicyclo[3.2.1]octane-3-yl)piperidine-4-carboxylic acid benzyl ester (intermediate 10) (22.0 g, 100.0%). The crude product was used without further purification.

[0238] LCMS (System 2, Method B): (ESI)m / z 487[M+H] + RT 3.10min, 202nm.

[0239]

[0240] Step 8: 1-((1R,3r,5S)-8-(3-(trifluoromethyl)-1,2,4-oxadiazol-5-yl)-8-azabicyclo [3.2.1] Synthesis of benzyl octane-3-yl)piperidine-4-carboxylate (intermediate 11)

[0241] TFA (110 mL, 5 vol.) was added dropwise to a DCM (220 mL) solution of 1-((1R,3r,5S)-8-((((tert-butoxycarbonyl)amino)oxy)(imino)methyl)-8-azabicyclo[3.2.1]octan-3-yl)piperidin-4-carboxylic acid benzyl ester (intermediate 10) (22.0 g, 45.3 mmol) at 0 °C to 5 °C. The reaction mixture was stirred at room temperature for 40 min, then cooled to 0 °C to 5 °C. TFAA (28.7 mL, 203.7 mmol) was then added dropwise and stirred for 30 min, then the mixture was heated to room temperature and stirred for 16 h. The reaction mixture was diluted with toluene (220 mL) and concentrated under reduced pressure. A saturated NaHCO3 solution (800 mL) was added and the reaction was extracted with ethyl acetate (3 × 500 mL). The combined organic layers were dried (Na2SO4) and concentrated under reduced pressure. The residue was purified by column chromatography [normal phase (neutral Al2O3), 0-30% (ethyl acetate in hexane)] to give 1-((1R,3r,5S)-8-(3-(trifluoromethyl)-1,2,4-oxadiazol-5-yl)-8-azabicyclo[3.2.1]octane-3-yl)piperidine-4-carboxylic acid benzyl ester (intermediate 11) (12.4 g, 59.0%).

[0242] LCMS (System 2, Method B): (ESI)m / z 465[M+H] + RT 4.40 min, 240 nm.

[0243]

[0244] Step 9: 1-((1R,3r,5S)-8-(3-(trifluoromethyl)-1,2,4-oxadiazol-5-yl)-8-azabicyclo [3.2.1] Synthesis of octane-3-yl)piperidine-4-carboxylate (intermediate 12).

[0245] Lithium hydroxide monohydrate (2.78 g, 66.3 mmol) was added in portions to a solution of 1-((1R,3r,5S)-8-(3-(trifluoromethyl)-1,2,4-oxadiazol-5-yl)-8-azabicyclo[3.2.1]octane-3-yl)piperidin-4-carboxylate (intermediate 11) (12.3 g, 26.5 mmol) in THF (123 mL) and water (24.6 mL) at room temperature, and the mixture was stirred at room temperature for 16 hours. The reaction mixture was diluted with water (100 mL), cooled to 0-10 °C, and the pH was adjusted to pH 5-6 using 1 M HCl solution (approximately 70-80 mL). The reaction mixture was stirred for 1 h, 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]octane-3-yl)piperidine-4-carboxylate (intermediate 12) (15.1 g, crude product). The crude product was used without further purification.

[0246] LCMS (System 2, Method B): (ESI)m / z 375[M+H] + RT 2.10min, 236nm.

[0247]

[0248] Step 10: N-(tert-butyl)-1-((1R,3r,5S)-8-(3-(trifluoromethyl)-1,2,4-oxadiazol-5-yl)-8- Synthesis of azabicyclo[3.2.1]octane-3-yl)piperidine-4-carboxamide (intermediate 14)

[0249] HATU (12.2 g, 32.0 mmol) was added in portions to a solution of 1-((1R,3r,5S)-8-(3-(trifluoromethyl)-1,2,4-oxadiazol-5-yl)-8-azabicyclo[3.2.1]octane-3-yl)piperidine-4-carboxylate (intermediate 12) (5.8 g, 14.1 mmol) (using 8 g of crude product separated from step 9) in DMF (80.0 mL) at 0 °C to 10 °C. The reaction mixture was stirred at 0 °C to 10 °C for 40 min. Then, 2-methylpropyl-2-amine (CAS: 75-64-9, intermediate 13) (6.8 mL, 64.2 mmol) and DIPEA (11.4 mL, 64.2 mmol) were added. The resulting reaction mixture was heated to room temperature and stirred for 16 h. Ice-cold water (500 mL) was added, and the mixture was stirred for 20 min. The precipitate formed was collected by filtration. The filter cake was washed with cold water (500 mL) and then with hexane (500 mL) to obtain a crude product (2.5 g). The aqueous layer was extracted with ethyl acetate (2 × 200 mL), and the combined organic layers were dried (Na₂SO₄) and concentrated under reduced pressure to obtain a further crude product (4.5 g). The crude products were combined and purified by column chromatography [normal phase (silica), 0-50% (ethyl acetate in hexane)] to obtain two batches of product with different purities (2.0 g and 3.8 g). Each batch was then taken 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.). 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]octane-3-yl)piperidine-4-carboxamide (intermediate 14) (1.9 g, 31.4%).

[0250] LCMS (System 2, Method B): (ESI)m / z 430[M+H] + RT 3.54 min, 240 nm.

[0251] 1H NMR (400MHz, DMSO-d6): δ7.31(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).

[0252]

[0253] Step 11: N-(tert-butyl)-1-((1R,3r,5S)-8-(3-(trifluoromethyl)-1,2,4-oxadiazol-5-yl)-8- Synthesis of aziridine-3-yl)piperidine-4-carboxamide hydrochloride (Example 1-1).

[0254] A solution of 1,4-dioxane in HCl (4 M, 19.0 mL, 10 vol.) was added 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]octane-3-yl)piperidine-4-carboxamide (intermediate 14) (1.9 g, 4.4 mmol) in 1,4-dioxane (3.8 mL). The reaction was stirred at room temperature for 4 hours, and then concentrated under reduced pressure. The resulting residue was azeotropically reacted with 1,4-dioxane (2 × 10 mL), and then ground 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) and 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]octane-3-yl)piperidine-4-carboxamide hydrochloride (Example 1-1) (1.8 g, 87.9%).

[0255] LCMS (System 2, Method B): (ESI)m / z 430[M+H] + RT 3.55min, 240nm.

[0256] 1 H NMR (400MHz, DMSO-d6): δ10.78–10.24(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).

[0257] N-(tert-butyl)-1-((1R,3r,5S)-8-(3-(trifluoromethyl)-1,2,4-oxadiazol-5-yl)-8-azadi Scale-up synthesis of cyclo[3.2.1]octane-3-yl)piperidine-4-carboxamide monohydrochloride monohydrate (Examples 1-2)

[0258]

[0259]

[0260] Step 1: Synthesis of cyclopent-3-ene-1-carboxylic acid benzyl ester (2)

[0261]

[0262] At room temperature (22-25°C), under a nitrogen atmosphere, add 2-methyl-THF (2550 mL), cyclopent-3-en-1-carboxylic acid (150 g), EDC·HCl (308.72 g), and benzyl alcohol (137.25 g) to a clean, dry 5.0 L four-necked RB flask equipped with a condenser and thermometer insert. Cool the reaction mixture to 0-5°C. At 0-5°C, add DMAP (196.10 g) and triethylamine (205 mL) to the reaction mixture over 5 minutes. Slowly heat the reaction mixture to room temperature and stir continuously at room temperature (24-25°C) for 18-20 hours. Monitor the reaction progress by TLC and HPLC until completion. Add demineralized water (3000 mL) at room temperature (24-25°C) and stir the mixture at room temperature (24-25°C) for 30 minutes. Separate the organic layer by adding 1N HCl (1500 mL) at 0-5℃ and stirring for 30 minutes. Separate the organic layer again by adding 10% NaHCO3 aqueous solution (1500 mL) at room temperature (24-25℃) and stirring for 15-20 minutes. Separate the organic layer once more by adding a brine solution (750 mL) at room temperature (22-25℃) and stirring for 15-20 minutes. Separate the organic layer again by vacuum distillation at 45-47℃ to obtain the crude compound.

[0263] Crude product weight: -248g, crude product yield: 91%, crude product properties: -violet liquid

[0264] The crude product is used directly in the next step without further purification.

[0265] Step 2: Synthesis of benzyl 6-oxabicyclo[3.1.0]hexane-3-carboxylate (15):

[0266]

[0267] At room temperature (22-25°C), under a nitrogen atmosphere, add MTBE (2450 mL) and benzyl cyclopentadienyl-1-carboxylate (Int-2, 245 g) to a clean, dry 5.0 L four-necked RB flask equipped with a condenser and thermometer insert, and cool the reaction mixture to 5-10°C. Add m-CPBA (362.6 g) in five separate batches, during which the color changes from dark brown to yellow. Slowly heat the reaction mixture to 15-20°C and continuously stir at 15-20°C for 16-20 hours. Monitor the reaction progress by HPLC until completion. Slowly add 20% sodium bisulfite aqueous solution (3675 mL, 15V) over 15-20 minutes and stir at 20-25°C for 30 minutes. Separate the organic layer and add 10% Na₂CO₃ aqueous solution (2450 mL, 10V), and stir at 20-25°C for 15 minutes. Separate the organic layer and add 20% sodium bisulfite aqueous solution (2450 mL, 10V) and stir until the peroxide content is <3 mg / L. Use a rapid peroxide detection strip. Separate the organic layer, dry it with sodium sulfate, filter it, and distill it off under pressure at 37-40℃ to obtain benzyl 6-oxabicyclo[3.1.0]hexane-3-carboxylate.

[0268] Crude product weight: -234g, crude product yield: -88%, crude product properties: -pale yellow liquid

[0269] The crude product is used directly in the next step without further purification.

[0270] Step-03: Synthesis of benzyl 4-oxo-2-(2-oxoethyl)butyrate (4):

[0271]

[0272] At room temperature (22-25°C), under a nitrogen atmosphere, ethyl acetate (380 mL) and periodic acid (87.41 g) were added to a clean, dry 2.0 L four-necked RB flask equipped with a condenser and thermometer insert, and the white suspension was cooled to 0-10°C. Benzyl 6-oxabicyclo[3.1.0]hexane-3-carboxylate (Int-15, 76 g, 380 mL ethyl acetate solution) was added, and the color was observed to change from dark brown to yellow. The reaction mixture was slowly heated to 15-20°C and continuously stirred for 3-4 hours. The reaction progress was monitored by TLC and HPLC until completion. Demineralized water (760 mL, 10V) was added, and the mixture was stirred three times at 20-25°C for 15-20 minutes. The organic layer was separated and washed with brine (380 mL, 5V) while stirring at 20-25°C for 5-10 minutes. The organic layer was separated, dried with sodium sulfate, filtered, and then distilled off under pressure at 37-40℃ to obtain benzyl 4-oxo-2-(2-oxoethyl)butyrate.

[0273] Crude product weight: -82g, Crude product properties: -Pale yellow liquid

[0274] The crude product is used directly in the next step without further purification.

[0275] Step-04: (1R,3R,5S)-3-(4-((benzyloxy)carbonyl)piperidin-1-yl)-8-azabicyclo[3.2.1]octyl Synthesis of tert-butyl alkyl-8-carboxylate (6):

[0276]

[0277] At room temperature (22-23°C), under a nitrogen atmosphere, add 570 mL of 2-methyl-THF and benzyl 4-oxo-2-(2-oxoethyl)butyrate (Int-4, 57 g * actual crude product weight 82 g, directly from step 3). Add Int-5 (internal amine) (49.54 g), and cool the reaction to 5-10°C, stirring for 30 minutes. Add sodium triacetoxyborohydride (56.78 g) and glacial acetic acid (5.7 mL), and heat the reaction mixture to room temperature (22-23°C), stirring for 10-12 hours. Monitor the reaction progress by TLC and HPLC, and after completion, add 400 mL of saturated bicarbonate solution (7V) and stir at 20-25°C for 15-20 minutes. The organic layer was separated, dried with sodium sulfate, filtered, and then distilled off under pressure at 37-40℃ to obtain (1R,3R,5S)-3-(4-((benzyloxy)carbonyl)piperidin-1-yl)-8-azabicyclo[3.2.1]octane-8-carboxylic acid tert-butyl ester.

[0278] Crude product weight: -115g, crude product properties: -thick liquid, color: -light chestnut. Purification using oxalate formation:

[0279] Add the crude product (25 g) and acetone (150 mL, 6V) to a clean, dry 500 mL 4-necked RB flask equipped with a thermometer and condenser, and cool to 5-10 °C. Add oxalic acid (10 g, 2.0 eq). Heat the reactants to room temperature while stirring for 2 hours. Distill off the reactants at 40 °C to obtain a crude product weighing -35 g. Then, add 10 vol. MTBE at 25-30 °C and stir for 30 minutes. Filter the reactants and wash the bed with MTBE (1.0 vol.) to obtain a wet solid (oxalate) weighing -26 g. Suspend the solid in a saturated bicarbonate solution (30 vol., 780 mL) and ethyl acetate (25 vol., 650 mL) and stir for 15 minutes. Separate the organic layer, dry with Na₂SO₄, and distill off the organic layer under vacuum at 37-40 °C to obtain the product. Product weight: -8.0 g.

[0280] Optionally, the substance can be further purified by column chromatography using hexane:ethyl acetate, adsorbed onto neutral alumina (Source-SDFCL). Column gradient: 5->10->12% ethyl acetate:hexane.

[0281] Step-05: 1-((1R,3R,5S)-8-azabicyclo[3.2.1]octane-3-yl)piperidine-4-carboxylic acid benzyl ester hydrochloride Synthesis of salt (7):

[0282]

[0283] Cyclopentyl methyl ether (452 ​​mL, 4.0 V) and Int-6 (113 g, 1.0 eq.) were added to a clean, dry 5.0 L 4-necked RB flask equipped with a condenser and thermometer under a nitrogen atmosphere. The reaction mixture was cooled to 0–5 °C. A solution of 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 heated to 22–25 °C and stirred for 16 hours. The reaction was monitored by HPLC until completion. The reaction mixture was filtered under a nitrogen atmosphere, the bed was washed with MTBE (2.0 vol.), the wet filter cake of the product was unloaded under nitrogen, 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.

[0284] Step 06: 1-((1R,3R,5S)-8-cyano-8-azabicyclo[3.2.1]octane-3-yl)piperidin-4-carboxylic acid benzyl Synthesis of ester (8):

[0285]

[0286] At 22-25°C, under a nitrogen atmosphere, dichloromethane (1860 mL, 10 vol.), Int-7 (186.0 g, 1.0 eq.), and triethylamine (355 mL, 5.0 eq.) were added over a period of 10-15 minutes to a clean, dry 5.0 L 4-necked RB flask equipped with a condenser and thermometer insert. The reaction mixture was stirred further for 30-45 minutes. The reaction mixture was cooled to 0-5°C, and a solution of cyanogen bromide (92 g, 1.7 eq.) in DCM (372 mL, 2.0 V) was added at 0-5°C. The reaction mixture was then slowly heated to 22-25°C and stirred at 22-25°C for 3-4 hours. The reaction progress was monitored by HPLC until completion. The reaction mixture was diluted with DCM (1860 mL, 10 vol.) and alkalized 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 / hexane. The pure fraction was collected and concentrated under reduced pressure to give 75 g of pure compound (yield: 46%).

[0287] Step-07: 1-((1R,3R,5S)-8-((((tert-butoxycarbonyl)amino)oxy)(imino)methyl)-8-nitrogen Synthesis of heterobicyclic [3.2.1]octane-3-yl)piperidine-4-carboxylic acid benzyl ester (10):

[0288]

[0289] 2-MethylTHF (800 mL, 10 vol.) and Int.8 (80 g, 1.0 eq.) were added to a clean, dry 3.0 L four-necked RB flask equipped with a condenser and thermometer under a nitrogen atmosphere, and the reaction mixture was cooled to 0–5 °C. ZnCl2 solution (2.2 eq., 1.9 M 2-methyl-THF solution) was added, and the reaction mixture was stirred at 0 °C for 30 min. Then, N-Boc-hydroxyamine (Int-9) (1.2 eq.) was added, and the reaction mixture was slowly heated to 23–25 °C with continuous 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 extracted with ethyl acetate (2 × 800 mL). The combined organic layers were dried over Na2SO4 and evaporated under reduced pressure to give 110 g (quantitative yield) of crude compound. This product was used directly in the next step without any further purification.

[0290] Step-08: 1-((1R,3R,5S)-8-(3-(trifluoromethyl)-1,2,4-oxadiazol-5-yl)-8-azabicyclo [3.2.1] Synthesis of benzyl octane-3-yl)piperidine-4-carboxylate (11):

[0291]

[0292] DCM (1150 mL, 10.0 vol.) and Int-10 (115.0 g, 1.0 eq.) were added to a clean, dry 3.0 L four-necked RB flask equipped with a condenser and thermometer insert 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 separate batches at 3–4 h intervals under a nitrogen atmosphere at 0–5 °C. The reaction mixture was stirred at room temperature for 12 h, then stirred for another 16 h until the reaction was complete (monitored by TLC and HPLC). After the reaction was complete, the reaction mixture was cooled to 0 °C, and the pH was adjusted to 7–8 using a saturated aqueous solution of 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 Na2SO4 and evaporated under reduced pressure to obtain 145g of crude compound, which was purified by column chromatography using neutral alumina with hexane and ethyl acetate as eluents to obtain 49g of pure product.

[0293] Step-09: 1-((1R,3R,5S)-8-(3-(trifluoromethyl)-1,2,4-oxadiazol-5-yl)-8-azabicyclo [3.2.1] Synthesis of octane-3-yl)piperidine-4-carboxylic acid (12):

[0294]

[0295] At room temperature (22-25°C), under a nitrogen atmosphere, THF (2250 mL), demineralized water (450 mL), and Int. 11 (225 g) were added to a clean, dry 5.0 L 4-necked RB flask equipped with a condenser and thermometer insert. 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 hours. The reaction progress was monitored by TLC and HPLC. After the reaction was complete, the reaction mixture was cooled to 0-5°C. The pH was adjusted to 4-5 using 1N HCl solution, and the mixture was stirred for 15 minutes. The solvent was evaporated, and the crude product was purified once MTBE (10.0 V) was added to the crude solid compound at room temperature and stirred at room temperature for 2 hours. The solid was filtered and washed with MTBE (1.0 V). The solid compound was dried under vacuum to obtain 245g of product (*the actual compound was 216g of Int-12, with the remaining approximately 29.0g being lithium chloride), which was a grayish-white solid.

[0296] Step-10: N-(tert-butyl)-1-((1R,3r,5S)-8-(3-(trifluoromethyl)-1,2,4-oxadiazol-5-yl)- Synthesis of 8-azabicyclo[3.2.1]octane-3-yl)piperidine-4-carboxamide

[0297]

[0298] program:

[0299] Acetonitrile (20.0 V) and Int-12 (1.0 eq.) were added in a single batch to a clean, dry 3-necked 1L round-bottom flask equipped with a mechanical stirrer and a nitrogen inlet, and the reaction mixture was stirred for 10 minutes. 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 a period of up to 15 minutes. After the addition was complete, the reaction mixture was stirred at room temperature for 16 hours. The reaction progress was monitored by TLC, and once complete, excess acetonitrile was distilled off under reduced pressure. The reaction was quenched with demineralized water (40.0 V), precipitating a solid, which was then stirred for 60 minutes. 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 a grayish-white solid.

[0300] Step-11: N-(tert-butyl)-1-((1R,3r,5S)-8-(3-(trifluoromethyl)-1,2,4-oxadiazol-5-yl)- Synthesis of 8-azabicyclo[3.2.1]octane-3-yl)piperidine-4-carboxamide hydrochloride

[0301]

[0302] A solution of Int-14 free base (56.0 g, 1.0 eq.) in acetone (10.0 V) was added to a flask, and the reaction mixture was stirred for 10 minutes. The reaction mixture was then cooled to 0 °C, and a 15% HCl IPA solution was added dropwise at 0 °C. The reaction mixture was then heated to 25–30 °C and stirred at 25–30 °C for 2 hours. The solvent was completely evaporated under reduced pressure, and the mixture was co-distilled twice with n-heptane (10.0 V). The solid compound was dried under vacuum to give 59.0 g of the product as a grayish-white solid.

[0303] Step 12: Recrystallization: N-(tert-butyl)-1-((1R,3r,5S)-8-(3-(trifluoromethyl)-1,2,4-oxadiazole- The synthesis of 5-yl)-8-azabicyclo[3.2.1]octane-3-yl)piperidine-4-carboxamide monohydrochloride monohydrate (Examples 1-2) become.

[0304] A solution of Int-14 HCl salt (108.0 g) in IPA (10.0 V) was added to a flask, and the reaction mixture was heated under reflux for 1 h; however, the solid was not completely soluble at reflux temperature. MeOH (1.0 V) was added, and heating was continued for 1 h to obtain a solution. The hot solution was passed through filter paper to remove insoluble particles. The solvent was removed under reduced pressure (approximately 8 volumes were removed from 11 volumes of solvent). The remaining approximately 3 volumes of solvent were then cooled to 0 °C and stirred at the same temperature for 1 h. The solid was then filtered and washed with a minimal amount of cooled IPA and dried by suction. The solid compound was dried under vacuum to give 93.5 g of product as a grayish-white solid.

[0305] Analysis was performed to determine the composition of the batch. The conclusion was that the batch was a monohydrochloride monohydrate.

[0306] Chemical formula of monohydrochloride monohydrate: C 20 H 33 Molecular weight of N5O3ClF3: 483.96

[0307]

[0308] Bioactivity

[0309] Example A: Determination of Phosphoric Acid-ERK1 / 2

[0310] Functional assays were performed using the Alphascreen Surefire phosphate-ERK1 / 2 assay (Crouch & Osmond, Comb. Chem. HighThroughput Screen, 2008). ERK1 / 2 phosphorylation is a downstream result of Gq / 11 and Gi / o protein-coupled receptor activation, making it well-suited for assessing M1, M3 (Gq / 11-coupled) and M2, M4 (Gi / o-coupled) receptors, rather than using different assays for different receptor subtypes. CHO cells stably expressing human muscarinic M1, M2, M3, or M4 receptors were plated (25K / well) in MEM-α + 10% dialyzed FBS on 96-well tissue culture plates. Once adhered, cells were serum-starved overnight. Agonist stimulation was performed for 5 min (37°C) by adding 5 μL of agonist to the cells. The culture 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 assay mixture was added. The plate was incubated in the dark with gentle shaking for 2 hours, and then read on a PHERAstar plate reader. pEC was calculated from the data obtained for each receptor subtype. 50 and E max The numerical results are listed in Table 1 below.

[0311]

[0312] Example B: CLint (in vitro hepatocytes) (Example 1)

[0313] Hepatocyte stability was determined using bioreclamation-preserved cryopreserved hepatocytes. Test compounds prepared in DMSO were incubated with hepatocytes at 37°C at a cell density of 1.0 million cells / mL with an initial concentration of 1 μM (final concentration 0.25% DMSO, n=2). Aliquots were removed at 0.5, 5, 10, 15, 30, 60, and 120 minutes to terminate the reaction, and the compounds were extracted with acetonitrile containing an analytical internal standard (0.5 μM carbamazepine). The samples were centrifuged, and the supernatant fraction of the parent compound was analyzed by mass spectrometry (LC-MS / MS). The amount of remaining compound (expressed as %) was determined from the MS response in each sample relative to the MS response in the T=0 sample (normalized to the internal standard). The half-life of compound disappearance was determined using an Ln plot of the remaining % using the following relationship:

[0314] Half-life (min) = -0.693 / λ (where λ is the slope of the Ln remainder % curve with respect to time).

[0315] The in vitro intrinsic clearance rate (CLint), expressed in μL / min / million cells, was calculated using the following formula:

[0316] Clint (μL / min / million cells) = (0.693 / half-life (min)) x (1000 / million cells per mL of incubation)

[0317] Mice = 7 uL / min / million

[0318] Rats 8uL / min / million

[0319] Dog 8uL / min / million

[0320] Monkeys <5uL / min / million

[0321] Humans <5uL / min / million

[0322] Example C: MDCK Permeability / Effusion (Example 1)

[0323] MDR1-MDCK cells (Solvo Biotechnology) were cultured at 2.35 × 10⁻⁶. 5 Cells / well were seeded onto 24-well Transwell plates and cultured at 37°C and 5% CO2 for 3 days before being used in confluent monolayers. For cell types, test and control compounds (propranolol, vincristine) (final 1 μM, 0.1% DMSO, n=2) were added to the donor compartment of the Transwell plate assembly in assay buffer (Hanks balanced salt solution supplemented with 25 mM HEPES, adjusted to pH 7.4) for top-to-base-outer (A>B) and base-outer to-top (B>A) measurements. Incubation was performed at 37°C, with samples removed from the donor and recipient compartments at T=0 and 1 hour, and compounds analyzed by mass spectrometry (LC-MS / MS) including analytical internal standards.

[0324] The apparent permeability (Papp) value is determined by the following relationship:

[0325] Papp = (Compound acceptor T = End / (Compound donor × V donor) / Incubation time) × V donor area × 60 × 10 -6 cm / s

[0326] Where V is the volume of each Transwell compartment (125 μL at the top, 600 μL at the base), and concentration is the relative MS response of the compound in the donor compartment (normalized to internal standard) before incubation and the compound in the recipient compartment at the end of incubation. Area = area of ​​cells exposed to drug transfer (0.33 cm²). 2The efflux ratio (Papp B>A / Papp A>B) was calculated from the average Papp value in each direction. The MDR1-MDCK cell line was engineered to overexpress the efflux transporter MDR1 (P-glycoprotein), and good permeability (B>A) but poor permeability (A>B) was observed, indicating that the compound is a substrate for this transporter. Fluorescein (LY) was added to the top buffer in all wells to assess cell layer viability. High LY transport indicates poor cell layer integrity because LY cannot freely permeate lipophilic barriers, and LY Papp > 10 × 10⁻⁶ indicates poor cell layer integrity. -6 Wells with a density of cm / s were rejected. It was noted that integrity failure in one well did not affect the validity of other wells on the plate. The recovery of compounds from the wells was determined by the MS responses (normalized to internal standard) in the donor and acceptor chambers at the end of incubation, compared to the MS response in the donor chamber before incubation. Recovery <50% indicates poor solubility, stability, or binding of the compound in the assay, which can reduce the reliability of the results.

[0327] AB = 66 × 10 -6 cm / sec

[0328] BA = 77 × 10 -6 cm / sec

[0329] BA / AB outflow ratio=1.2

[0330] Example D: Solubility Data (Example 1)

[0331] Water-soluble (thermodynamic)-LCMS / MS method

[0332] Prepare a 10 mM stock solution (in DMSO) for the test sample. From the 10 mM stock solution, prepare a 1 μM working solution by diluting the test sample in the mobile phase solution (typically methanol: 2 mM ammonium acetate containing a suitable internal standard (IS) – carbamazepine / any other suitable IS). Additionally, serially dilute the working solution in the mobile phase solution to up to 5 to 6 linear points to prepare standard solutions for plotting calibration curves. Analyze the area of ​​each standard sample in a single peak using LCMS / MS. Plot the normalized area values ​​against concentration to obtain a calibration equation to identify unknown samples. To determine the thermodynamic (TD) water solubility of the test compound, add 1 mg (in powder form) of the compound to 1 mL of each of the buffers and biorelevant media mentioned in the table below to obtain a theoretical concentration equivalent to 1 mg / mL. Disperse the test compound in the buffer solution using a vortex mixer.

[0333] Sr No. Reagent Name

[0334] 1 SGF pH–1.2

[0335] 2. Blank FaSSIF (Aq. buffer) pH–6.5

[0336] 3 FaSSIF pH–6.5

[0337] The resulting solution was then incubated on a RotoSpin at 50 rpm for 4 hours to determine TD solubility at room temperature (25°C). After the incubation period, the solution was filtered using a 0.45 μm PVDF syringe filter to remove the insoluble fraction of the compound. The filtrate was diluted in the mobile phase, and the AUC of the diluted sample was subsequently determined using LCMS / MS. Based on the AUC of the tested sample, the corresponding concentration was calculated using a 5- to 6-point linearity / calibration curve.

[0338] All values ​​are reported in μM.

[0339]

[0340] Example E: HμREL (Example 1)

[0341] In HμREL Pool TM When the 96-well liver co-culture plate arrives, change the culture medium and allow the cells to acclimatize at 37°C for approximately 20 hours. Incubation medium (serum-free) and test compound (final substrate concentration 1 μM; final DMSO concentration 0.1%) were added to... A 96-well co-culture system (final cell count 30,000 cells / well) was used to initiate the reaction. The final incubation volume at each time point was 80 μL. Each assay included two control compounds. All incubations were performed individually for each test compound.

[0342] Each compound was incubated for 0, 2, 6, 24, 48, and 72 hours (0, 120, 360, 1440, 2880, and 4320 minutes). The reaction was terminated by transferring 60 μL of the incubator to 180 μL of acetonitrile containing an internal standard at the appropriate time point. The stop plate was centrifuged at 3000 rpm for 20 minutes at 4 °C to precipitate any residual protein.

[0343] Quantitative analysis

[0344] After protein precipitation, the sample supernatant was combined in a cassette containing up to four compounds and analyzed using Cyprotex universal LC-MS / MS conditions.

[0345] Data Analysis

[0346] From the graph of In peak area ratio (compound peak area / internal standard peak area) versus time, determine the slope of the line. Then, calculate the half-life (t) using the following equation. 1 / 2 ) and inherent clearance rate (CL) int ):

[0347] Elimination rate constant (k) = (-slope)

[0348]

[0349]

[0350] (Where V = incubation volume (μL) / number of cells)

[0351] CLint < 0.143 uL / min / million

[0352] Example F: Target recruitment for predicting effective human doses (Example 1)

[0353] The expected requirements for observing the efficacy of M1 agonists in humans are based on recombinant M1 EC. 50 Six hours of unbound brain exposure. The unbound brain exposure required to achieve the M1 agonist efficacy at a dose of approximately 22 mg in Example 1 was predicted, and the predicted human half-life was 15 hours. Figure 1 ).

[0354] Parameters used in receptor occupancy prediction (target engagement).

[0355] MW = 429.48

[0356] M1 pEC 50 =7.17

[0357] Fu = 0.682

[0358] Kpuu=1

[0359] Half-life (predicted) = 15h

[0360] F = 0.61

[0361] CL = 4.4 ml / min / kg

[0362] v = 5.9 ml / min / kg

[0363] Ka = 1

[0364] definition:

[0365] fu - the portion of unbound compounds in blood plasma or brain tissue homogenate.

[0366] F - Bioavailability; the percentage of the administered dose that reaches the systemic circulation (plasma).

[0367] Kp,uu - The ratio of unbound brain concentration to unbound plasma concentration. Quantifies the net flux of a drug across the blood-brain barrier, including the quantitative action of transport proteins, without being confounded by nonspecific binding in plasma and brain tissue. (Gupta et al., DMD, 2006; Hammarlund-Udenaes et al., PharmRes, 2008)

[0368] Example G: Subchronic PCP-induced in an operant reversal learning task in female Lister Hooded rats Defect reduction (Example 1)

[0369] Objectives and Results

[0370] The ability of Example 1 (1, 3, 10, and 30 mg / kg, po, 1-hour pretreatment time ptt) to alleviate the disruption of cognitive tasks induced by subchronic treatment with benzo[a]pyridine (scPCP) was investigated in female Lister Hooded rats.

[0371] Compared to the media group, the scPCP group showed a significantly lower percentage of correct responses during the task reversal phase (P<0.01). Figure 2 Compared to the scPCP group in the reversal phase, the group treated with Example 1 at the lowest and two intermediate doses (1, 3, and 10 mg / kg) significantly (P<0.05, P<0.05, and P<0.01, respectively) achieved a significantly higher percentage of correct response. Figure 2 ).

[0372] Materials and methods

[0373] Female Lister-hooded rats were used in this experiment. The average weight of the rats at the time of testing was 294 g ± 29 g. Rats were divided into 3–5 groups and housed under standard laboratory conditions with a 12-hour light:dark cycle (light at 07:00) and food restricted to 90% of their free-feeding body weight (12 g food per rat per day). Testing was conducted during the light phase. Rats were randomly assigned to two treatment groups and received either the medium, n = 8 (0.9% saline solution, ip) or PCP, n = 48 (2 mg / kg, ip, twice daily for 7 days). On the day of testing, rats were randomly assigned to 7 treatment groups (n = 6–8 per group) to receive acute treatment with either Example 1 or the medium (1, 3, 10, and 30 mg / kg, po, 1-hour ptt). Example 1 was dissolved in 1% methylcellulose and administered orally at a volume of 5 ml / kg one hour prior to testing (po). The study was conducted in accordance with the Animal Science Procedures Act (UK, 1986) and approved by the University of Manchester AWERB (Animal Welfare and Ethics Review Board).

[0374] Experimental Procedure

[0375] After acclimatizing to the operating room, rats were trained to respond to food in a FR1 (fixed ratio 1) reinforcement program with both levers active. Once the response stabilized, rats were trained to press either the left or right lever for food delivery, with the active lever varying daily. Each phase lasted 20 minutes, and counts were recorded for each lever. Rats were then trained to respond to food based on the location of a visual cue (a lit LED). Half of the rats were trained to press the lever under the lit LED to receive a food reward, while the other half received the opposite randomness training (pressing the lever under a non-lit LED). The experimental phase ended after a total of 128 lever presses, which took approximately 30 minutes. Rats were then trained until they again met the opposite randomness criteria.

[0376] One day prior to each reversal learning task phase, a full 30-minute operant training phase (as described above) is conducted to ensure stable responses. For the reversal learning task, animals are first exposed for 5 minutes during which chance (the cue position relative to the active lever) is identical to that of the operant training phase. Responses on both the correct and incorrect levers are recorded during this period. This portion of the phase is referred to as the initial phase. In the subsequent 5-minute period, chance is reversed. Responses again on both the correct and incorrect levers are recorded. This second period is referred to as the reversal phase. In this phase, training is terminated, and rats are treated with PCP (2 mg / kg, ip, or 0.9% saline, ip) for 7 days, followed by a washout period of at least 7 days. Rats are randomized such that all rats in the cage receive different drug treatments.

[0377] Data are expressed as percentage of correct response (±SEM), with values ​​for the initial and reversal phases given for different drug treatment groups. Figure 2 The percentage of correct responses was used to determine whether the drug had a significant effect on response accuracy (e.g., potentially reflecting cognitive impairment); statistical significance was assumed when P < 0.05, and one-way ANOVA was used to determine statistical significance to detect the primary effect of drug treatment in the initial and reversal phases. The total number of lever presses recorded during the initial and reversal phases after treatment with the medium or Example 1 was not significantly different (Table 2), confirming no nonspecific effect on the overall response in this study.

[0378] Table 2. Effects of acute treatment with Example 1 (1.0, 3.0, 10.0, and 30.0 mg / kg, po) on overall performance in the reversal learning task in rats treated with scPCP (2 mg / kg, ip, twice daily for 7 days, followed by a washout period of at least 7 days). Data are presented as mean total number of lever presses ± SEM (n = 6–9) during the initial and retention phases of the reversal learning task.

[0379] Drug treatment initial stage Reversal phase SC saline + medium 27.0±0.2 27.3±0.2 scPCP+Vehicle 26.5±0.2 26.0±0.5 scPCP+ Example 1 1.0 mg / kg 27.4±0.2 27.5±0.3 scPCP+ Example 1 3.0 mg / kg 27.5±0.2 26.8±0.2 scPCP+ Example 1 10.0 mg / kg 27.0±0.3 26.6±0.5 scPCP+ Example 1 30.0 mg / kg 26.4±0.6 26.6±0.3

[0380] Brief description of the attached figures

[0381] Figure 1 : Figure 1 The predicted target recruitment for the effective dose of Example 1 is shown, derived from allometric scaling in mouse, rat, dog, and monkey species following oral administration. The data are presented as a function of the calculated M1 receptor target recruitment % where 50% is equivalent to recombinant human EC50. 50 Unbound exposure (86 nM or 28 ng / ml). Based on the measured equivalent unbound plasma: brain distribution map (Kpuu = 1), the exposure shown represents exposure in plasma or ventricles.

[0382] Figure 2 : Figure 2 The effect 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 was shown in rats treated with scPCP (2 mg / kg, ip, twice daily for 7 days, followed by a washout period of at least 7 days). Data are presented as mean correct response % ± SEM (n = 6–9). Dashed lines separate the initial phase (left) and reversal phase (right) of the task. Data were analyzed by one-way ANOVA followed by LSD test. ***P < 0.001; % correct response was significantly lower in the reversal phase of the task compared to the sc saline + mediator group. #P < 0.05; ##P = 0.01; ###P < 0.001; % correct response was significantly increased in the reversal phase of the task compared to the scPCP + mediator group.

[0383] equivalent

[0384] The above embodiments are provided to illustrate the purpose of the present invention and should not be construed as imposing any limitation on the scope of the invention. It is obvious that various modifications and changes can be made to the specific embodiments of the invention exemplified above and in the embodiments without departing from the principles of the invention. All such modifications and changes are intended to be included in this application.

Claims

1. Compound of formula (1): (1)； Or its salt.

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

3. A pharmaceutically acceptable salt of the compound according to claim 1.

4. The acid addition salt of the compound according to claim 1.

5. The hydrochloride salt of the compound according to claim 1.

6. The monohydrochloride salt of the compound according to claim 1.

7. The compound according to claim 1, wherein it is a compound of formula (2): (2)； Or its salt.

8. A salt of the compound according to claim 7.

9. A pharmaceutically acceptable salt of the compound according to claim 7.

10. The acid addition salt of the compound according to claim 7.

11. The hydrochloride salt of the compound according to claim 7.

12. The monohydrochloride salt of the compound according to claim 7.

13. The compound according to claim 1, wherein it is a compound of formula (2b): (2b)。 14. The compound according to claim 1, wherein it is a compound of formula (2c): (2c)。 15. The compound according to claim 1, wherein it is a compound of formula (2): (2)。 16. A pharmaceutical composition comprising the compound of claim 1 or 7 or a salt thereof; or a salt of any one of claims 2 to 6 or 8 to 14; or the compound of claim 15; and a pharmaceutically acceptable excipient.

17. Use of the compound of claim 1 or 7 or a salt thereof, or a salt of any one of claims 2 to 6 or 8 to 14, or a compound of claim 15, or a pharmaceutical composition of claim 16, in the preparation of a medicament for treating an individual's disorder selected from cognitive disorders and psychotic disorders.

18. The use according to claim 17, wherein the impairment is a cognitive impairment.

19. The use according to claim 17 or 18, wherein the cognitive impairment is a cognitive impairment associated with Alzheimer's disease.

20. The use according to claim 17 or 18, wherein the cognitive impairment is a cognitive impairment associated with Parkinson's disease.

21. The use according to claim 17, wherein the obstacle is Alzheimer's disease.

22. The use according to claim 17, wherein the obstacle is Lewy body dementia.

23. The use according to claim 17, wherein the disorder is schizophrenia.

24. Use of the compound of claim 1 or 7 or a salt thereof, or a salt of any one of claims 2 to 6 or 8 to 14, or a compound of claim 15, or a pharmaceutical composition of claim 16, in the preparation of a medicament for treating an individual’s acute pain, chronic pain, neuropathic pain, or inflammatory pain, or for reducing the severity of an individual’s acute pain, chronic pain, neuropathic pain, or inflammatory pain.

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