Nitrogen-containing heterocyclic compound having a phenyl group and its uses

Abstract

The present invention relates to a nitrogen-containing heterocyclic compound having a phenyl group and its use.The compound according to the present invention can effectively improve synaptic plasticity without side effects, thereby exerting therapeutic effects on various neurological disorders such as neurodegenerative diseases, neurodegenerative pathologies, neuropsychiatric disorders, memory loss, cognitive dysfunction / injury, and extinction learning disorders.

Description

[Technical field]

[0001] The present invention relates to a nitrogen-containing heterocyclic compound having a phenyl group and its use. More specifically, the present invention relates to a compound, composition or method for the prevention or treatment of neurological or neurodegenerative disorders. [Background technology]

[0002] Synaptic plasticity and critical periods

[0003] Neural networks are networks of neurons. Synapses are nodes in neural networks where neurons exchange information with each other. Within a network, synapses are "plastic" because their number and strength change continuously throughout life as synaptic inputs are added, removed, or refined in response to ongoing neural activity. Synaptic plasticity refers to this neural activity-dependent transformation of synapses, manifested by changes in the strength and / or efficiency of synaptic transmission.

[0004] Higher functions such as learning, cognition, synthesis, judgment, art, and creativity depend on the ability to form neural networks in the cerebral cortex (called cortical neural networks). Synaptic plasticity of cortical neural networks reaches a peak early after birth, known as the critical period. After the critical period, cortical synaptic plasticity gradually decreases with age, eventually reaching a state where experience no longer actively changes the cerebral cortex. Interestingly, however, recent studies have shown that under certain conditions, synaptic plasticity can be restored to a high level even after the critical period has ended: in animal models, peripheral nerve injury reactivated synaptic plasticity in the somatosensory thalamocortical circuits that transmit peripheral sensory signals to the cerebral cortex (Yu et al., 2012, Neuron. 74:731-42; Petrus et al., 2014, Neuron. 81:664-73; Gao et al., 2014, J Neurosci. 34:10770-9).

[0005] Reduced synaptic plasticity is also associated with the degeneration of adult brain function. In particular, synaptic plasticity in thalamocortical circuits is greatly restricted with age. As mentioned above, if it is possible to resume the critical period, reactivating or increasing cortical synaptic plasticity through therapeutic means may be useful for treating diseases caused by reduced synaptic plasticity.

[0006] Histone deacetylase inhibitors

[0007] Most of the DNA in cells exists packaged in a more dense, compacted form known as chromatin. Compaction occurs by wrapping DNA around a group of proteins called histones. The degree of compaction of chromatin depends on the level of acetylation of the histones. In general, histone acetyltransferases (HATs) perform the acetylation that allows access to DNA, and histone deacetylases (HDACs) perform the deacetylation that blocks access to DNA. Compaction of chromatin acts as a regulatory mechanism for gene expression, since transcription can occur only if access is permitted (Verdin and Ott, 2015, Nat Rev Mol Cell Biol 16:258-264).

[0008] To date, 11 types of mammalian HDACs have been identified. These types are broadly divided into four classes based on the degree of homology with yeast HDACs. Class I includes HDACs 1, 2, 3, and 8. Class II HDACs are divided into class IIa (HDACs 4, 5, 7, and 9) and class IIb (HDACs 6 and 10) based on structural similarity. Class III HDACs include SIRT1-7, and class IV includes HDAC11.

[0009] Class I HDACs are primarily restricted to the nucleus, whereas class II HDACs shuttle between the nucleus and the cytoplasm through phosphorylation of specific serine residues. The N-terminal domain of HDACs interacts with multiple tissue-specific transcription factors and acts as a transcription inhibitor. The C-terminal domain of HDACs possesses deacetylase enzymatic activity. However, class IIa HDACs do not possess deacetylase activity because this class has a loss-of-function mutation in which the tyrosine (Tyr) residue in the C-terminal catalytic domain is replaced by histidine (His).

[0010] HDAC4

[0011] HDAC4 is expressed throughout the body. HDAC4 is particularly highly expressed in the brain, heart and skeletal muscle (Grozinger et al., 1999, Proc. Natl. Acad. Sci. USA 96, 4868-4873; Darcy et al., 2010, J. Comp. Neurol. 518, 722-740). In the brain, HDAC4 levels peak immediately after birth (Sando et al., 2012, Cell 151, 821-834). HDAC4 was detected in corticotropin-releasing hormone (CRH) neurons (100±0%), oxytocin neurons (98±2%), vasopressin neurons (100±0%), orexin neurons (100±0%), AgRP neurons (100±0%), POMC neurons (92±4%), histamine neurons (85±7%), dopamine neurons (100±0%), serotonin neurons (93±2%), and noradrenaline neurons (100±0%) (reference [Takase et al., 2013, PloS ONE 8:e58473]).

[0012] Cytoplasmic HDAC4 is rapidly translocated to the nucleus in response to low potassium or excitotoxic glutamate conditions that induce apoptosis. Ectopic overexpression of HDAC4 in the nucleus promotes neuronal cell death and inhibits the transcriptional activity of survival factors such as MEF2 and CREB (Bolger and Yao, 2005, J Neurosci. 2005; 25:9544-9553). Neurons containing HDAC4 are homogeneous in size and widely distributed in the neuropil, including the paraventricular nucleus (PVN), lateral hypothalamus (LHA), hypothalamic arcuate nucleus (ARC), tuberous mammary nucleus (TMN), dorsal raphe (DR), and locus coeruleus (LC). Notably, HDAC4 colocalizes with PSD95 (Mielcarek et al., 2013, PLoS Biol. 11:e1001717; Takase et al., 2013, PloS ONE 8:e58473), suggesting a role for HDAC4 in synaptic plasticity (Sando et al., 2012, Cell 151, 821-834; Mielcarek et al., 2013, PLoS Biol. 11:e1001717).

[0013] In animal models, tissue-specific deletion of HDAC4 in the rat hippocampus eliminated long-lasting stress-induced behavioral changes (Sailaja et al., 2012, Proc. Natl. Acad. Sci. USA 109, E3687-E3695). Overexpression of HDAC4 accelerated the death of cerebellar granule neurons by increasing their sensitivity to H2O2 injury through inhibition of peroxisome proliferator-activated receptor α (PPARα) activity (Bolger and Yao, 2005, J. Neurosci. 25, 9544-9553; Li et al., 2012, Nat. Med. 18, 783-790; Sando et al., 2012; Cell 151, 821-834; Yang et al., 2011, FEBS Lett. 585, 761-766). Overexpression of HDAC4 in the rat hippocampus induced depression-like behaviors (Sarkar et al., 2014, Neuropsychopharmacology 39, 2221-2232).

[0014] In a Drosophila model, overexpression of HDAC4 in adult mushroom body neurons, a structure important for memory formation, caused impairment of long-term courtship memory but did not affect short-term memory (Fitzsimons et al., 2013, PloS ONE 8: e83903). Subsequent studies investigating the relationship with the subcellular distribution of HDAC4 showed that deficits in mushroom body morphogenesis, eye development, and long-term memory were associated with nuclear HDAC4 abundance (Main et al., 2021, Front. Mol. Neurosci. 14:616642).

[0015] In humans, increased levels of HDAC4 have been identified in postmortem HD frontotemporal lobar degeneration (FTLD) brains (Yeh et al., 2013, Brain Res. 1504, 16-24; Whitehouse et al., 2014, Neuropathol. Appl. Neurobiol. 41, 245-257). HDAC4 has been identified as a component of Lewy bodies in Parkinson's disease brains and as a component of intranuclear inclusions in neuronal intranuclear inclusion disease (Takahashi-Fujigasaki and Fujigasaki, 2006, Neuropathol. Appl. Neurobiol. 32, 562-566). HDAC4 has been observed to be significantly overexpressed in specific cortical regions of autistic patients (Nardone et al., 2014, Transl. Psychiatry 4:e433).

[0016] HDAC inhibitors and synaptic plasticity

[0017] Histone modifications have been proposed as a potential way to restore cortical plasticity in adulthood. Valproic acid is known as a histone deacetylase (HDAC) inhibitor. In a clinical study in which participants took valproic acid capsules, the critical period was reopened and absolute pitch learning, which was only possible during the critical period, became possible in adults (Gervain et al., 2013, Front Syst Neurosci. 7:102). When adult rats were exposed to an enriching environment compared to normal cage conditions, the total amount of H3 acetylation in the visual cortex increased compared to the control group, and ocular dominance plasticity, which only appears during the critical period, was also observed after the critical period (Baroncelli L et al., 2016, J Neurosci. 36:3430-40).

[0018] Treatment with the HDAC inhibitor SAHA (suberoylanilide hydroxamic acid) increased H3 acetylation in the visual cortex to levels typically observed in young rats, particularly at the P3 promoter of brain-derived neurotrophic factor (BNDF), known to be important for developmental synaptic plasticity and long-term memory in the visual cortex (Baroncelli L et al., 2016, J Neurosci. 36:3430-40). Increased BDNF expression was also observed in neurospheres (Bagheri et al., 2019, Int. J. Mol. Sci. 20: 1109).

[0019] SAHA or other HDAC inhibitors have been shown to be effective in ameliorating symptoms in mouse models of Huntington's disease (HD) (Ferrante et al., 2003; Hockly et al., 2003, J. Biol. Chem. 278, 16059-16072; Guardian et al., 2005, J. Biol. Chem. 280, 556-563). HDAC4 correlates with the length of polyglutamine expressed in HD. It is colocalized with cytoplasmic inclusions and a wide range of peripheral organ pathologies such as skeletal muscle atrophy and heart failure manifested in Huntington's disease (Zielonka et al., 2014, Front. Physiol. 5:380; Mielcarek et al., 2014, PLoS Genet. 10:e1004550; Mielcarek et al., 2014, PloS ONE 9:e108961; Zielonka et al., 2014, Exp. Clin. Cardiol. 20, 2547-2554). Reducing HDAC4 levels in the R6 / 2 mouse model of Huntington's disease delayed the formation of cytoplasmic aggregates in various regions of the brain, improved motor coordination and the neurological phenotype, and extended lifespan (Mielcarek et al., 2013, PLoS Biol. 11:e1001717).

[0020] Interestingly, SAHA treatment in R6 / 2 mice restored cortical transcript levels of BDNF (Mielcarek et al., 2011, PloS ONE 6:e27746). It has been shown that HDAC4 and HDAC5 can inhibit specific BDNF transcripts, and this effect is reversed by SAHA treatment, so the memory enhancing and neuroprotective effects of HDAC inhibitors may be associated with increased BDNF expression (Koppel and Timmusk, 2013, Neuropharmacology 75, 106-11).

[0021] HDAC4 can play a central role in brain physiology and can be a useful target for treating various neurodegenerative disorders. However, currently available HDAC inhibitors inhibit HDACs broadly, which makes such inhibitors unsuitable for inhibiting specific HDACs. SAHA increases synaptic plasticity, but causes serious side effects including fatigue, nausea, diarrhea, or thrombocytopenia, which makes SAHA unsuitable for human use (reference [Meghan E. et al., 2011, Bone. 48: 1117-1126]).

[0022] Therefore, it would be useful to develop selective HDAC inhibitors for targeted therapy. Summary of the Invention [Problem to be solved by the invention]

[0023] An object of the present invention is to solve all of the above problems.

[0024] An object of the present invention is to provide a nitrogen-containing heterocyclic compound having a phenyl group, a stereoisomer thereof, or a pharma- ceutical acceptable salt thereof for improving synaptic plasticity.

[0025] Another object of the present invention is to provide a pharmaceutical composition for treating or preventing a neurological or neurodegenerative disorder mediated by HDAC, comprising as an active ingredient a nitrogen-containing heterocyclic compound having a phenyl group for improving synaptic plasticity, a stereoisomer thereof, or a pharma- ceutically acceptable salt thereof.

[0026] Another object of the present invention is to provide a method for treating or preventing a neurological or neurodegenerative disorder mediated by HDAC, comprising administering to a subject a nitrogen-containing heterocyclic compound having a phenyl group, a stereoisomer thereof, or a pharma- ceutically acceptable salt thereof for improving synaptic plasticity.

[0027] Another object of the present invention is to provide a pharmaceutical composition for combined administration containing a nitrogen-containing heterocyclic compound having a phenyl group for improving synaptic plasticity, a stereoisomer thereof, or a pharma- ceutically acceptable salt thereof and a second therapeutic agent, and a combined administration method using the same.

[0028] The object of the present invention is not limited to the above object. The object of the present invention will become clearer in the following description and will be realized by the means and combinations thereof described in the claims. [Means for solving the problem]

[0029] A representative configuration of the present invention for achieving the above object is as follows.

[0030] According to one aspect of the present invention, there is provided a compound of formula (1), a stereoisomer thereof, or a pharma- ceutically acceptable salt thereof:

[0031] JPEG2025512899000002.jpg4495

[0032] In the above formula, R 1 is an aryl or heteroaryl, or combines with an adjacent phenyl group to form a saturated or partially saturated heterocycle, and one or more hydrogen atoms of the aryl, heteroaryl, or heterocycle are unsubstituted or substituted with a halogen atom, C 1-6 Alkyl, C 3-12 Cycloalkyl, C 1-6 alkoxy, aryl, heteroaryl, amino, or nitro, the substituents being unsubstituted or substituted with one or more halogen atoms or C 1-6 is substituted with alkyl,

[0033] R 2 is hydrogen, C 1-6 alkyl or aryl;

[0034] R 3is hydrogen, carboxyl, N-hydroxycarboxamide, or carboxamide, and the hydrogen in said group is C 1-6 may be substituted with alkyl;

[0035] n is 0 or 1,

[0036] m is 0 to 6.

[0037] In one embodiment, R 1 is phenyl, pyridinyl, pyridazinyl, pyrimidinyl, pyrazinyl, furanyl, thiophenyl, pyrrolyl, imidazolyl, or oxazolyl, or can be joined with an adjacent phenyl group to form quinoxalinyl or dihydrobenzodioxinyl.

[0038] In other embodiments, R 1 Of which, one or more hydrogen atoms are halogen atoms, one or more halogen-substituted or unsubstituted C 1-6 Alkyl or C 3-12 Cycloalkyl, C 1-6 Alkoxy, phenyl, pyridinyl, pyridazinyl, pyrimidinyl, one or more C 1-6 It may be substituted with alkyl or unsubstituted amino, or with nitro.

[0039] In one embodiment, R 1may be phenyl, biphenyl, pyridin-4-yl, pyridin-3-yl, pyridin-2-yl, methylpyridin-3-yl, 6-methoxy-pyridin-3-yl, 6-cyclopropyl-pyridin-3-yl, trifluoromethylpyridin-3-yl, phenylpyridin-3-yl, pyridazin-4-yl, pyridazin-3-yl, pyridazin-2-yl, pyrimidin-2-yl, pyrimidin-4-yl, pyrimidin-5-yl, methylpyrimidin-5-yl, 5-fluoropyrimidin-2-yl, 2-(trifluoromethyl)pyrimidin-5-yl, 2-cyclopropylpyrimidin-5-yl, pyrazin-2-yl, thiophen-2-yl, furan-2-yl, oxazol-2-yl, 3-nitrophenyl, 4-nitrophenyl, 3-aminophenyl, 4-aminophenyl, or 4-diethylaminophenyl.

[0040] In one embodiment, R 2 is H or C 1-6 It may be alkyl.

[0041] In one embodiment, R 3 may be H, carboxyl, methylcarboxyl, ethylcarboxyl, N-hydroxycarboxamide, or carboxamide.

[0042] In one embodiment, the stereoisomers may be optical isomers.

[0043] In one embodiment, the stereoisomer may be the (3R,6S)-enantiomer.

[0044] In one embodiment, the compound may be a compound selected from the compounds presented in Tables 2 and 3, a stereoisomer thereof, or a pharma- ceutically acceptable salt thereof.

[0045] In other embodiments, the compound is BnH12003, BnH12017, BnH12045, BnH12046, BnH12047, BnH12048, BnH12051, BnH12053, BnH12057, BnH12058, BnH12064, BnH12066, BnH12067, BnH1207 1, BnH12072, BnH12078, BnH12085, BnH12086, BnH12088, BnH12089, BnH12090, BnH12091, BnH12092, BnH12093, BnH12095, BnH12097, and BnH12098.

[0046] In one embodiment, the compound may be an inhibitor of HDAC, hi another embodiment, the HDAC may be HDAC4.

[0047] In one embodiment, the compound may increase synaptic plasticity.

[0048] In accordance with yet another aspect of the present invention, there is provided a pharmaceutical composition comprising the compound, a stereoisomer thereof, or a pharma- ceutically acceptable salt thereof as an active ingredient.

[0049] In one embodiment, the composition may be for the prevention or treatment of a neurological or neurodegenerative disorder.

[0050] In other embodiments, the compound, its stereoisomer, or a pharma- ceutically acceptable salt thereof can be delivered to the brain at a concentration of about 0.1 to about 3 μM.

[0051] In another embodiment, the disorder may be Alzheimer's disease.

[0052] In another embodiment, the disorder may be PTSD.

[0053] In one embodiment, the composition can further comprise a second therapeutic agent.

[0054] In other embodiments, the second therapeutic agent may be solanezumab.

[0055] According to yet another aspect of the present invention, there is provided a method for preventing or treating a neurological or neurodegenerative disorder, comprising administering the compound, a stereoisomer thereof, or a pharma- ceutically acceptable salt thereof to an individual in need thereof.

[0056] According to yet another aspect of the present invention there is provided the use of the compound, its stereoisomer, or a pharma- ceutically acceptable salt thereof for use as a medicament or for the manufacture of a medicament.

[0057] According to one embodiment, said use is for reactivating or increasing brain synaptic plasticity.

[0058] According to another embodiment, the use is for the prevention or treatment of a neurological or neurodegenerative disorder. Effect of the Invention

[0059] The compounds according to the present invention can effectively improve or reactivate synaptic plasticity without side effects, thereby exerting therapeutic effects on various neurological disorders, such as neurodegenerative diseases, neurodegenerative pathologies, neuropsychiatric disorders, memory loss, cognitive dysfunction / injury, and extinction learning disorders. [Brief description of the drawings]

[0060] [Figure 1a] 1 shows the results of measuring cell viability in SH-SY5Y cells in response to treatment with BnH12003 (0, 0.1, 1, 10, 100 μM).

[0061] [Figure 1b] 1 shows the results of measuring cell viability in 293T cells in response to treatment with BnH12003 (0, 0.1, 1, 10, 100 μM).

[0062] [Figure 2a] The figure shows the site of osmotic pump (Os pump) injection in a mouse experiment.

[0063] [Figure 2b] The patch recording sites in the mouse experiments are shown.

[0064] [Figure 3a] The results of measuring the excitation / inhibition balance in mouse barrel cortex (layer 4) in response to administration of BnH12003 (Os pump, ventricular injection, 3 μM) are shown.

[0065] [Figure 3b] Changes in TC EPSC potency in mouse barrel cortex (layer 4) in response to administration of BnH12003 (Os pump, ventricular injection, 3 μM) are shown.

[0066] [Figure 4a] The results of measuring the excitation / inhibition balance in mouse barrel cortex (layer 4) in response to intraperitoneal administration of BnH12003 (25 mg / kg) are shown.

[0067] [Figure 4b] 1 shows changes in TC EPSC potency in mouse barrel cortex (layer 4) in response to intraperitoneal administration of BnH12003 (25 mg / kg).

[0068] [Diagram 5] The results of measuring changes in HDAC4 protein expression in mouse barrel cortex (layer 4) in response to administration of BnH12003 (Os pump, injected into barrel cortex) are shown.

[0069] [Figure 6] 1 shows the degree of symptom improvement in a rodent (rat) model of post-traumatic stress disorder in response to intraperitoneal administration of BnH12003 (25 mg / kg).

[0070] [Figure 7a] Changes in the input / output curve (I / O curve) of field excitatory postsynaptic potential (fEPSP) in the hippocampus of an Alzheimer's disease mouse model in response to administration of BnH12003 and solanezumab (Os pump; 3 μM, 200 μg).

[0071] [Figure 7b] 1 shows the results of LTP measurements of field excitatory postsynaptic potentials in the hippocampus of an Alzheimer's disease mouse model in response to administration of BnH12003 and solanezumab (Os pump; 3 μM, 200 μg).

[0072] [Figure 8a] The results of measuring the excitation / inhibition balance in the barrel cortex (layer 4) of an Alzheimer's disease mouse model in response to administration of BnH12003 and solanezumab (lateral ventricle, Os pump; 3 μM, 200 μg) are shown.

[0073] [Figure 8b] 1 shows changes in TC EPSC potency in the barrel cortex (layer 4) of an Alzheimer's disease mouse model in response to administration of BnH12003 and solanezumab (Os pump; 3 μM, 200 μg).

[0074] [Figure 9] 1 shows the binding characteristics of BnH12003 to HDAC4.

[0075] [Figure 10a] 1 shows the knockdown effect of HDAC4 shRNA on HDAC4, BDNF, and GluN2B expression in the human cell line SH-SY5Y. [Figure 10b] 1 shows the knockdown effect of HDAC4 shRNA on HDAC4, BDNF, and GluN2B expression in the human cell line SH-SY5Y.

[0076] [Figure 11a] The effect of BnH12003 on the expression of BDNF and GluN2B is shown. [Figure 11b] The effect of BnH12003 on the expression of BDNF and GluN2B is shown. [Figure 11c] The effect of BnH12003 on the expression of BDNF and GluN2B is shown.

[0077] [Figure 12a] The time course effect of BnH12003 on HDAC4 expression is shown. [Figure 12b] The time course effect of BnH12003 on HDAC4 expression is shown. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0078] The detailed description of the present invention below will be described with reference to certain drawings and with respect to specific embodiments in which the present invention may be practiced, but the present invention is not limited thereto, and is limited only by the appended claims together with the full scope of equivalents to which such claims are entitled when properly described. Terms presented in describing the present invention should generally be understood in their ordinary sense unless otherwise indicated, and apply to all of the same terms used herein, as well as to the aspects or embodiments of the invention in which the terms are defined.

[0079] Throughout the detailed description of the present invention, when compositions and kits are described as having or comprising specific components, it is contemplated that there are further compositions and kits of the present invention that consist essentially of or consist of the recited components.

[0080] In applications where an element or component is referred to as being included / included or selected from a list of mentioned elements or components, it should be understood that the element or component can be any one of the mentioned elements or components, or that the element or component can be selected from the group consisting of the mentioned elements or components.

[0081] It should be understood that the order of steps or sequence for performing certain operations is immaterial so long as the invention remains operable. Moreover, two or more steps or operations may be performed simultaneously.

[0082] At various places in the specification, variables or parameters are disclosed as groups or ranges, and the detailed description is specifically intended to include all individual members of such groups and ranges.

[0083] In this specification, examples or exemplary language, such as "such as" or "including," are merely intended to better describe the invention and do not limit the scope of the invention unless claimed. Nothing in the specification should be construed as indicating any non-claimed element as essential to the practice of the invention.

[0084] I. Definition

[0085] The singular terms are used herein to refer to the plural (ie, to at least one) unless the context requires otherwise.

[0086] "And / or" is used herein to mean "and" or "or," unless otherwise indicated.

[0087] "At least one" includes each and every referenced object after the expression individually, unless otherwise understood from context and usage, and various combinations of two or more of the referenced objects.

[0088] "Comprise," "including," "has," "having," "containing," "including" (including grammatical equivalents thereof) should generally be understood to be open-ended and non-limiting, e.g., not excluding additional, unrecited elements or steps unless otherwise specifically stated or understood from the context.

[0089] "About" means approximately, in the vicinity, roughly, or to the extent of. When the term "about" is used in conjunction with a numerical range, it modifies that range by extending the boundaries around the stated numerical values. In general, the term "about" is used herein to modify a numerical value around the stated value by a variance of 10%.

[0090] The terms "individual," "patient," and "subject" are used interchangeably and include any animal, including mammals (e.g., mice, rats, other rodents, rabbits, dogs, cats, pigs, cows, sheep, horses, or primates, including humans).

[0091] "Cure," "treat," or "treatment" includes any effect that results in the improvement of a disease, illness, disorder, etc., such as alleviating, reducing, modulating, ameliorating, relieving, or eliminating, or ameliorating the symptoms thereof. Treatment may be curing, improving, or at least partially reversing the disorder. In certain embodiments, treatment is curing the disease.

[0092] "Active agent" is used to refer to a compound having biological activity or a pharma- ceutically acceptable salt thereof. In some embodiments, an "active agent" is a compound having pharmaceutical utility or a pharma-ceutically acceptable salt thereof. For example, an active agent may be an anti-neurodegenerative therapeutic agent.

[0093] "Compound" means a chemical entity having a structure depicted by a chemical formula set forth herein, a stereoisomer thereof (including a mixture of stereoisomers), or a pharma- ceutically acceptable salt thereof.

[0094] "Pharmaceutically acceptable" includes molecular entities and dosage forms that do not elicit adverse, allergic or other abnormal reactions when administered appropriately to animals or humans.

[0095] "Pharmaceutically acceptable salts" refers to those salts that retain the biological effects and properties of the free base or free acid. For example, pharmaceutically acceptable salts can be prepared by adding an inorganic or organic base to the free acid. Salts derived from inorganic bases include, but are not limited to, sodium, potassium, lithium, ammonium, calcium, magnesium salts, and the like. Salts derived from organic bases include, but are not limited to, primary, secondary, and tertiary amines, substituted amines including naturally occurring substituted amines, cyclic amines, and basic ion exchange resins, such as isopropylamine, trimethylamine, diethylamine, triethylamine, tripropylamine, ethanolamine, diethylamine, lysine, arginine, N-ethylpiperidine, piperidine, piperazine, and the like.

[0096] "Pharmaceutically acceptable excipient" and "pharmaceutically acceptable carrier" refer to substances that aid in the administration and absorption of an active agent by a subject, and can be included in the compositions of the present invention without causing significant adverse toxicological effects to the patient. Non-limiting examples of pharmaceutically acceptable excipients include water, NaCl, saline, lactated Ringer's, normal sucrose, normal glucose, binders, fillers, disintegrants, lubricants, coatings, sweeteners, flavorings, salt solutions (e.g., Ringer's solution), alcohols, oils, gelatin, carbohydrates such as lactose, amylose or starch, fatty acid esters, hydroxymethylcellulose, polyvinylpyrrolidine, coloring agents, and the like. Such preparations can be sterilized and, if desired, can be mixed with auxiliary substances that do not adversely react with the compounds of the present invention, such as lubricants, preservatives, stabilizers, wetting agents, emulsifiers, salts that affect osmotic pressure, buffers, coloring agents, and / or aromatic substances. Those skilled in the art will recognize that other pharmaceutical excipients are useful in the present invention.

[0097] Exemplary pharma- ceutically acceptable carriers or components thereof are sugars, such as lactose, glucose, and sucrose; starches, such as corn starch and potato starch; cellulose and its derivatives, such as sodium carboxymethylcellulose, ethylcellulose, and methylcellulose; powdered tragacanth; malt; gelatin; talc; solid lubricants, such as stearic acid and magnesium stearate; calcium sulfate; synthetic oils; vegetable oils, such as peanut oil, cottonseed oil, sesame oil, olive oil, and corn oil; polyols, such as propylene glycol, glycerin, sorbitol, mannitol, and polyethylene glycol; alginic acid; phosphate buffers; emulsifiers, such as TWEENs(r); wetting agents, such as sodium lauryl sulfate; colorants; flavorings; tableting; stabilizers; antioxidants; preservatives; pyrogen-free water; isotonic saline; and phosphate buffers.

[0098] A "pharmaceutical composition" or "pharmaceutical dosage form" refers to a combination of an active agent with an inert or active carrier.

[0099] An "effective amount" refers to an amount of a compound or composition (e.g., a compound or composition of the present invention) sufficient to achieve a beneficial or desired result. An effective amount can be administered in one or more administrations, applications, or dosages, and is not intended to be limited to a particular dosage form or route of administration.

[0100] "Administering" refers to oral administration, administration as a suppository, topical contact, intravenous, parenteral, intraperitoneal, intramuscular, intralesional, intraspinal, intracranial, intraventricular, intranasal, or subcutaneous administration to a subject, or implantation of a sustained release device, such as a mini-Os pump. Administration is by any route, including parenteral and transmucosal (e.g., buccal, sublingual, palatal, gingival, nasal, vaginal, rectal, or transdermal). Parenteral administration includes, for example, intravenous, intramuscular, intraarterial, intradermal, subcutaneous, intraperitoneal, intraventricular, and intracranial administration. Other modes of delivery include, but are not limited to, the use of liposomal dosage forms, intravenous infusion, transdermal patches, and the like. "Co-administration" or "administration in combination" refers to administration of a composition described herein simultaneously with, immediately prior to, or immediately following the administration of one or more additional therapies (e.g., anti-cancer agents, chemotherapeutic agents, or agents for treating neurodegenerative diseases). The compounds of the present invention can be administered alone or co-administered to a patient. Co-administration is intended to include simultaneous or sequential administration of compounds individually or in combination (one or more compounds or formulations). Thus, formulations can also be combined with other active agents if desired (e.g., to reduce metabolic degradation).

[0101] The pharmaceutical dosage forms of the present invention can be administered to mammals, e.g., humans, but also to other mammals, e.g., animals in need of veterinary treatment, such as livestock (e.g., dogs, cats, etc.), farm animals (e.g., cows, sheep, pigs, horses, etc.), and laboratory animals (e.g., rats, mice, guinea pigs, etc.).

[0102] The "cognitive function" of a subject can be defined as intellectual activity or function. Examples of intellectual activity or function include, but are not limited to, attention, processing speed, learning and memory, executive function, verbal fluency, and working memory. For example, the composition of the present invention can improve cognitive function if it improves one or more intellectual activities or functions in a subject with impaired cognitive function.

[0103] "Histone deacetylase" or "HDAC" is intended to refer to any member of a family of enzymes that remove Nε-acetyl groups from the ε-amino groups of lysine residues of proteins (e.g., histones or tubulin). Unless the context indicates otherwise, the term "histone" refers to any histone protein, including H1, H2A, H2B, H3, H4, and H5 from any species.

[0104] An "HDAC-mediated disease or disorder" or a "histone deacetylase-mediated disease or disorder" refers to a disease or disorder in which HDAC and / or the action of HDAC is important or necessary, for example, for the onset, progression or development.

[0105] "HDAC enzymatic activity inhibition" or "HDAC inhibition" means decreasing the catalytic activity of HDAC or decreasing an activity of HDAC that is unrelated to the catalytic activity.

[0106] "Neurological disorders" include neurodegenerative diseases, neuropsychiatric disorders, memory loss, cognitive impairment / damage, and extinction learning disorders.

[0107] "Neurodegenerative disease" means any disorder that can be reversed, prevented, managed, treated, ameliorated, or eliminated by a formulation that stimulates the generation of new neurons.

[0108] "Memory" refers to the ability to retrieve information about past events or knowledge.

[0109] "Age-related memory loss" refers to any continuum of conditions characterized by a deterioration of neurological function that does not rise to the level of dementia, as further defined herein and / or in the literature [Diagnostic and Statistical Manual of Mental Disorders: 4th Edition of the American Psychiatric Association (DSM-IV, 1994)].

[0110] "Injury-related memory loss" refers to memory loss where there has been brain damage and possibly also neurological damage.

[0111] "Impaired cognitive function" refers to cognitive function that is not as robust as that observed in age-matched normal subjects, and includes states of cognitive decline.

[0112] As used herein, unless otherwise specified, "alkyl" generally refers to any alkyl group having a C1 to C6 carbon atom. 10 "refers to saturated linear, branched, or cyclic primary, secondary, or tertiary hydrocarbons of the formula (I) and specifically includes methyl, CF3, CCl3, CFCl2, CF2Cl, ethyl, CH2CF3, CF2CF3, propyl, isopropyl, cyclopropyl, butyl, isobutyl, sec-butyl, t-butyl, pentyl, cyclopentyl, isopentyl, neopentyl, hexyl, isohexyl, cyclohexyl, cyclohexylmethyl, 3-methylpentyl, 2,2-dimethylbutyl, and 2,3-dimethylbutyl. The term includes all substituted and unsubstituted alkyl groups, particularly halogenated alkyl groups, and more particularly fluorinated alkyl groups. Non-limiting examples of moieties with which the alkyl group may be substituted are selected from the group consisting of halogen (fluoro, chloro, bromo or iodo), hydroxyl, amino, alkylamino, arylamino, alkoxy, aryloxy, nitro, cyano, sulfonic acid, sulfate, phosphonic acid, phosphate, or phosphonate, optionally unprotected or protected, as known to those of skill in the art, e.g., as taught in Greene, et al., Protective Groups in Organic Synthesis, John Wiley and Sons, Second Edition, 1991, incorporated herein by reference.

[0113] As used herein, "aryl" refers to an aromatic monocyclic or polycyclic ring system containing 6 to 14 carbon atoms, unless otherwise specified, specifically including phenyl, biphenyl, or naphthyl, preferably phenyl. The term includes all substituted and unsubstituted moieties. The aryl group may be substituted with any of the described moieties, including but not limited to one or more moieties selected from the group consisting of halogen (fluoro, chloro, bromo, or iodo), hydroxyl, amino, alkylamino, arylamino, cycloalkyl, alkoxy, aryloxy, nitro, cyano, sulfonic acid, sulfate, phosphonic acid, phosphate, or phosphonate, unprotected or protected as appropriate, as known to those skilled in the art, for example, as taught in [Greene, et al., Protective Groups in Organic Synthesis, John Wiley and Sons, Second Edition, 1991], which is incorporated herein by reference.

[0114] As used herein, unless otherwise specified, "heteroaryl" refers to an aromatic monocyclic or polycyclic ring system having 5 to 14 ring atoms, one to four of which are independently O, N or S, and the remaining ring atoms are carbon atoms, and preferably an aromatic monocyclic ring system having 5 or 6 ring atoms, one to four of which are independently O, N or S, and the remaining ring atoms are carbon atoms, specifically including pyridinyl, pyridazinyl, pyrimidinyl, pyrazinyl, pyrrolyl, furanyl, thiophenyl, imidazolyl, or oxazolyl. Heteroaryl groups may be substituted with any of the described moieties, including, but not limited to, one or more moieties selected from the group consisting of halogen (fluoro, chloro, bromo, or iodo), hydroxyl, amino, alkylamino, arylamino, cycloalkyl, alkoxy, aryloxy, thiol, nitro, cyano, sulfonic acid, sulfate, phosphonic acid, phosphate, or phosphonate, optionally unprotected or protected, as known to those of skill in the art, for example, as taught in Greene, et al., Protective Groups in Organic Synthesis, John Wiley and Sons, Second Edition, 1991, which is incorporated herein by reference.

[0115] II. Active Compounds

[0116] In one aspect of the present invention, there is provided a compound of formula (1), a stereoisomer thereof, or a pharma- ceutically acceptable salt thereof:

[0117] JPEG2025512899000003.jpg4291

[0118] In the above formula,

[0119] R 1is an aryl or heteroaryl, or combines with an adjacent phenyl group to form a saturated or partially saturated heterocycle, and one or more hydrogen atoms of the aryl, heteroaryl, or heterocycle are unsubstituted or substituted with a halogen atom, C 1-6 Alkyl, C 3-12 Cycloalkyl, C 1-6 alkoxy, aryl, heteroaryl, amino, or nitro, the substituents being unsubstituted or again containing one or more halogen atoms or C 1-6 is substituted with alkyl,

[0120] R 2 is hydrogen, C 1-6 alkyl or aryl;

[0121] R 3 is hydrogen, carboxyl, N-hydroxycarboxamide, or carboxamide, and the hydrogen in said group is unsubstituted or C 1-6 may be substituted with alkyl;

[0122] n is 0 or 1,

[0123] m is 0 to 6.

[0124] In one embodiment, the saturated or partially saturated heterocycle may be a bicycle having 9 to 12 ring atoms, in which one or more (preferably one or two) of the ring atoms are independently O, N, or S, and the remaining ring atoms are carbon atoms.

[0125] In one embodiment, the stereoisomer may be an optical isomer. Preferably, the compound may be in the form of the (3R,6S)-enantiomer or the (3S,6R)-enantiomer.

[0126] In one embodiment, R 1is phenyl, pyridinyl, pyridazinyl, pyrimidinyl, pyrazinyl, furanyl, thiophenyl, pyrrolyl, imidazolyl, or oxazolyl, or can be joined with an adjacent phenyl group to form quinoxalinyl or dihydrobenzodioxinyl.

[0127] In one embodiment, R 1 At least one hydrogen atom is unsubstituted or is a halogen atom, C 1-6 Alkyl, C 3-12 Cycloalkyl, C 1-6 It may be substituted with alkoxy, aryl (preferably phenyl), heteroaryl (preferably pyridinyl, pyridazinyl, pyrimidinyl, pyrrolyl, furanyl, thiophenyl, imidazolyl or oxazolyl), amino or nitro, said substituents again being selected from halogen atoms and C 1-6 It may be substituted with one or more groups selected from alkyl.

[0128] In one embodiment, R 1 Of which, one or more hydrogen atoms are halogen atoms, one or more halogen-substituted or unsubstituted C 1-6 Alkyl or C 3-12 Cycloalkyl, C 1-6 Alkoxy, phenyl, pyridinyl, pyridazinyl, pyrimidinyl, one or more C 1-6 It may be substituted with alkyl or unsubstituted amino, or with nitro.

[0129] In one embodiment, R 1may be phenyl, biphenyl, pyridin-4-yl, pyridin-3-yl, pyridin-2-yl, methylpyridin-3-yl, 6-methoxy-pyridin-3-yl, 6-cyclopropyl-pyridin-3-yl, trifluoromethylpyridin-3-yl, phenylpyridin-3-yl, pyridazin-4-yl, pyridazin-3-yl, pyridazin-2-yl, pyrimidin-2-yl, pyrimidin-4-yl, pyrimidin-5-yl, methylpyrimidin-5-yl, 5-fluoropyrimidin-2-yl, 2-(trifluoromethyl)pyrimidin-5-yl, 2-cyclopropylpyrimidin-5-yl, thiophen-2-yl, furan-2-yl, oxazol-2-yl, 3-nitrophenyl, 4-nitrophenyl, 3-aminophenyl, 4-aminophenyl, or 4-diethylaminophenyl.

[0130] In one embodiment, R 1 may be substituted at the 3- or 4-position of the phenyl group.

[0131] In one embodiment, R 2 is H or C 1-6 It may be alkyl.

[0132] In one embodiment, R 3 may be H, carboxyl, methylcarboxyl, ethylcarboxyl, N-hydroxycarboxamide, or carboxamide.

[0133] In one embodiment, m may be 0 to 3. In another embodiment, m may be 0 or 1.

[0134] In one embodiment, the compound of formula (1) may be provided in the form of a pharma- ceutically acceptable salt thereof.Preferably, said salt may be a sodium salt.

[0135] In one embodiment, compounds of Formula (1) include, but are not limited to, those presented in Table 1 below:

[0136] [Table 1]

[0137] JPEG2025512899000005.jpg199145

[0138] JPEG2025512899000006.jpg197144

[0139] JPEG2025512899000007.jpg199145

[0140] JPEG2025512899000008.jpg34144

[0141]

[0142] In one example, the compounds and compositions described herein may be inhibitors of HDAC. Preferably, the HDAC is HDAC4.

[0143] In other embodiments, the compounds and compositions described herein are synaptic plasticity modulators. Preferably, the synaptic plasticity is synaptic plasticity in the barrel cortex (e.g., layer 4) of the brain.

[0144] III. Pharmaceutical Compositions

[0145] In general, the compounds described herein can be formulated into pharmaceutical compositions using techniques known to those skilled in the art. The concentration of the active compound in the pharmaceutical composition will vary depending on the absorption, inactivation, and excretion rates of the drug, as well as other factors known to those skilled in the art. It should be noted that dosage values ​​will also vary depending on the severity of the disease to be alleviated. It should be further understood that for any particular subject, the specific administration regimen should be adjusted over time according to the individual's needs and the professional judgment of the person administering or supervising the administration of the composition, and that the concentration ranges described herein are exemplary only and are not intended to limit the scope or practice of the claimed compositions. The active ingredient can be administered at once or divided into a number of smaller doses to be administered at various time intervals.

[0146] The concentration of an inhibitor that reduces HDAC activity to 50% of the activity of uninhibited HDAC is called the IC 50 In some embodiments, the reduction in HDAC activity is determined as an IC value of less than 100 nanomolar. In some embodiments, the reduction in HDAC activity is at least 50%, such as at least about 75%, such as at least about 90%. In some embodiments, the HDAC activity is reduced by at least 95%, such as at least 99%. In some embodiments, the compounds described herein have an IC value of less than 100 nanomolar. 50 In some embodiments, the compounds described herein have IC values ​​between 100 nanomolar and 1 micromolar. 50 In some embodiments, the compounds described herein have an IC value of 1 to 25 micromolar. 50 It has a value.

[0147] In some embodiments, such inhibition is specific, i.e., the histone deacetylase inhibitor reduces the ability of histone deacetylase to remove acetyl groups from proteins at a concentration lower than the concentration of the inhibitor required to also produce another unrelated biological effect. In some embodiments, the concentration of the inhibitor required for histone deacetylase inhibitory activity is at least 2-fold lower, e.g., at least 5-fold lower, e.g., at least 10-fold lower, e.g., at least 20-fold lower, than the concentration required to produce the unrelated biological effect.

[0148] In some embodiments, the pharmaceutical composition comprises about 25 mg, about 50 mg, about 75 mg, about 100 mg, about 150 mg, about 200 mg, about 250 mg, about 300 mg, about 350 mg, about 400 mg, about 450 mg, about 500 mg, about 600 mg, about 700 mg, about 800 mg, about 900 mg, about 1000 mg, about 1250 mg, about 1500 mg, about 1750 mg, about 2000 mg, about 2250 mg, about 2500 mg, about 2750 mg, or about 3000 mg of the compound.

[0149] In some embodiments, the pharmaceutical composition contains from about 0.01 mg to about 125 mg, preferably from about 1 mg to about 50 mg, per kg of subject body weight per day, one or more times per day to obtain the desired therapeutic effect.

[0150] In a specific embodiment, the composition contains, for example, 0.0001 to 99.9% by weight, preferably 0.001 to 50% by weight of the compound based on the weight of the total composition. The content ratio is a value based on the dry weight from which the solvent has been removed.

[0151] In certain embodiments, the composition comprises a 5% (w / v), 10% (w / v), 15% (w / v), 20% (w / v), 25% (w / v), 30% (w / v), 35% (w / v), or 40% (w / v) aqueous solution of one or more compounds. In some embodiments, an effective amount of the composition comprises a 25% (w / v) solution of one or more compounds.

[0152] In certain embodiments, the pharmaceutical composition of the present invention further comprises one or more pharma- ceutically acceptable excipients. In some embodiments, the one or more pharma- ceutically acceptable excipients are selected from the group comprising a diluent, a buffer, a preservative, a stabilizer, a solubilizer, or any combination thereof.

[0153] In certain embodiments, the pharmaceutical composition can be formulated for administration as a liquid dosage form suitable for oral, intracavity, intradermal, intramuscular, intraspinal, intravenous, subcutaneous, or intraventricular administration.

[0154] In certain embodiments, the liquid dosage form of the pharmaceutical composition as described herein further comprises a diluent. In some embodiments, the inert diluent is saline.

[0155] In certain embodiments, the liquid dosage form of the pharmaceutical composition as described herein further comprises a buffer.For example, buffers suitable for use in the present invention include all organic and inorganic acids and their salts, such as citrate buffers (e.g., monosodium citrate-disodium citrate mixture, citric acid-trisodium citrate mixture, citric acid-monosodium citrate mixture), succinate buffers (e.g., succinic acid-monosodium succinate mixture, succinic acid-sodium hydroxide mixture, succinic acid-disodium succinate mixture), tartrate buffers (e.g., tartaric acid-sodium tartrate mixture, tartaric acid-potassium tartrate mixture, tartaric acid-sodium hydroxide mixture), fumarate buffers (e.g., fumaric acid-monosodium fumarate mixture, fumaric acid-fumaric acid mixture), and the like. Examples of buffers include, but are not limited to, acetic acid-sodium acetate mixture, monosodium fumarate-disodium fumarate mixture, gluconate buffers (e.g., gluconic acid-sodium gluconate mixture, gluconic acid-sodium hydroxide mixture, gluconic acid-potassium gluconate mixture), oxalate buffers (e.g., oxalic acid-sodium oxalate mixture, oxalic acid-sodium hydroxide mixture, oxalic acid-potassium oxalate mixture, etc.), lactate buffers (e.g., lactic acid-sodium lactate mixture, lactic acid-sodium hydroxide mixture, lactic acid-potassium lactate mixture, etc.), and acetate buffers (e.g., acetic acid-sodium acetate mixture, acetic acid-sodium hydroxide mixture, etc.). Phosphate buffers, histidine buffers, and trimethylamine salts, such as Tris, can also be used.

[0156] In certain embodiments, the liquid dosage form of pharmaceutical composition as described herein further comprises preservative.For example, the preservative suitable for use in the present invention includes, but is not limited to, phenol, benzyl alcohol, meta-cresol, methylparaben, propylparaben, octadecyldimethylbenzylammonium chloride, benzalkonium halide (e.g., chloride, bromide, and iodide), hexamethonium chloride, and alkylparaben (e.g., methyl or propylparaben, catechol, resorcinol, cyclohexanol, and 3-pentanol).

[0157] In certain embodiments, the liquid dosage form of pharmaceutical composition as described herein further comprises stabilizer.For example, suitable stabilizer includes but is not limited to polyhydric sugar alcohol, trihydric or higher sugar alcohol, amino acid, organic sugar or sugar alcohol, polyvinylpyrrolidone monosaccharide, trisaccharide, polysaccharide, protein, sulfur-containing reducing agent, amino acid polymer, and polyethylene glycol.

[0158] In certain embodiments, the liquid dosage form of the pharmaceutical composition as described herein further comprises a solubilizer.In some embodiments, the solubilizer is an ionic surfactant.Examples of non-ionic surfactants include, but are not limited to, polysorbates, polyoxamers, pluronic polyols, and polyoxyethylene sorbitan monoethers.

[0159] In certain embodiments, the pharmaceutical compositions disclosed herein can be prepared for storage as lyophilized dosage forms or aqueous solutions by mixing the compositions with any pharma- ceutically acceptable carriers, excipients, or stabilizers typically used in the art (e.g., buffers, stabilizers, preservatives, isotonicity agents, non-ionic detergents, antioxidants, and various other additives).

[0160] Where the compound is administered orally, it can be formulated with a pharma-ceutically acceptable excipient in the form of a pill, capsule, cachet, tablet, etc.

[0161] Compressed tablets can be produced by compressing in a suitable machine the active ingredient in a free-flowing form such as a powder or granules, optionally mixed with a binder, lubricant, inert diluent, preservative, surfactant, or dispersant. Molded tablets can be produced by molding in a suitable machine a mixture of the powdered active ingredient moistened with an inert liquid diluent. The tablets can be optionally coated or scored, and are optionally formulated to provide slow or controlled release of the active ingredient. Tablets, troches, lozenges, aqueous or oil suspensions, dispersible powders or granules, emulsions, hard or soft capsules, e.g., gelatin capsules, syrups, or elixirs can be prepared for oral use. Dosage forms of the compounds of the present invention intended for oral use can be prepared by any method known in the art for the manufacture of pharmaceutical compositions, and such compositions can contain one or more ingredients, including sweeteners, flavorings, colorings, and preservatives, to provide a palatable preparation. The tablet containing the active ingredient is suitably mixed with non-toxic pharma- ceutically acceptable excipients suitable for tablet manufacture. Such excipients may be, for example, inert diluents such as calcium or sodium carbonate, lactose, calcium or sodium phosphate; granulating and disintegrating agents such as corn starch or alginic acid; binders such as starch, gelatin or acacia; and lubricants such as magnesium stearate, stearic acid or talc. Tablets may be coated by known techniques, including uncoated or microencapsulated, to delay disintegration and absorption in the gastrointestinal tract and provide a sustained action over a long period of time. For example, a time-delay material such as glyceryl monostearate or glyceryl distearate may be used alone or with a wax.

[0162] In certain embodiments, the pharmaceutical compositions herein are administered to a subject by, for example, oral, intracavity, intradermal, intramuscular, intraspinal, intravenous, subcutaneous, or intraventricular administration.

[0163] IV. Biological Assay

[0164] Methods for selecting compounds useful in the present invention are described herein.

[0165] In one embodiment, the compound of the present invention is selected for its ability to interact with HDAC.In one embodiment, HDAC is HDAC4.In another embodiment, the binding of the compound to HDAC is measured by surface plasmon resonance assay.In one embodiment, the compound is BnH12003.

[0166] In one embodiment, the compound is selected for its ability to increase the expression of BDNF and / or GluN2B in targeted neurons. In one embodiment, the compound is selected for its ability to increase the expression of BDNF in targeted neurons. In yet another embodiment, the compound is selected for its ability to increase the expression of GluN2B in targeted neurons. In one embodiment, RT-PCR is used to measure the expression level. In one embodiment, BnH12003, BnH12017, BnH12045, BnH12046, BnH12047, BnH12048, BnH12051, BnH12053, BnH12057, BnH12058, BnH12064, BnH12066, BnH12067, BnH12070, BnH12072, BnH12074, BnH12076, BnH12078, BnH12079, BnH12080, BnH12081, BnH12082, BnH12083, BnH12084, BnH12085, BnH12086, BnH12087, BnH12088, BnH12089, BnH12090, BnH12091, BnH12092, BnH12093, BnH12094, BnH12095, BnH12096, BnH12097, BnH12098, BnH12099, BnH20 ... H12071, BnH12072, BnH12078, BnH12085, BnH12086, BnH12088, BnH12089, BnH12090, BnH12091, BnH12092, BnH12093, BnH12095, BnH12097, or BnH12098. In other embodiments, the compound is BnH12003, BnH12046, BnH12057, BnH12067, BnH12091, BnH12092, BnH12095, BnH12097, or BnH12098.

[0167] In one embodiment, the compound of the present invention is selected for its non-toxicity to human cells. In one embodiment, the compound of the present invention is selected for its lack of significant effect on the cell viability of the human cell line SH-SY5Y. In yet another embodiment, the compound of the present invention is selected for its lack of significant effect on the cell viability of the human cell line 293T. In one embodiment, the compound does not reduce the cell viability of SH-SY5Y or 293T at a concentration of about 0.1, 1, 10, 100 μM. In one embodiment, the compound may be one selected from the compounds listed in Table 4. Preferably, the compound is BnH12003.

[0168] In one embodiment, the compounds of the invention are selected for their ability to increase potency as measured through TC EPSC potency in rodent barrel cortex (layer 4). In one embodiment, the compounds increase potency as measured at a concentration of about 0.1 to about 5 μM when delivered by the Os pump via ventricular infusion. In yet another embodiment, the concentration may be about 0.1 to about 3 μM. In yet another embodiment, the concentration may be about 3 μM. In yet another embodiment, the concentration is about 0.03, 0.1, 0.3, 1, 3, 10 mg / kg. In one embodiment, the compound is BnH12003. In yet another embodiment, the compound is delivered by intraperitoneal infusion, wherein the compound is BnH12003. In yet another embodiment, the intraperitoneal infusion delivers about 25 mg / kg of the compound.

[0169] In one embodiment, the compounds of the invention, alone or in combination with a monoclonal antibody against amyloid-β peptide, reduce the I / O gradient (ms s) of the field excitatory postsynaptic potential (fEPSP) in the hippocampus of rodent models of Alzheimer's disease. -1In one embodiment, the compound increases the I / O gradient when administered via the Os pump with a monoclonal antibody against amyloid beta peptide at a rate of about 3 μM and 200 μg for each compound and antibody. In yet another embodiment, the monoclonal antibody is solanezumab. In yet another embodiment, the compounds are BnH12003 and solanezumab.

[0170] In one embodiment, the compound of the present invention is selected for its ability to increase LTP of fEPSP gradient in hippocampus of rodent model of Alzheimer's disease in combination with monoclonal antibody against amyloid β peptide.In one embodiment, the compound increases LTP of fEPSP gradient when administered through Os pump with monoclonal antibody against amyloid β peptide at a rate of about 3 μM, 200 μg for each compound and antibody.In yet another embodiment, the monoclonal antibody is solanezumab.In yet another embodiment, the compound is BnH12003.

[0171] In one embodiment, the compounds of the present invention are selected for their ability to increase potency as measured via TC EPSC potency in the barrel cortex (layer 4) of a rodent model of Alzheimer's disease. In one embodiment, the compound increases potency measured at a concentration of about 3 μM when delivered by the Os pump via ventricular infusion. In yet another embodiment, the compound increases TC EPSC potency when the compound is delivered with a monoclonal antibody against amyloid β peptide. In yet another embodiment, the monoclonal antibody is solanezumab. In yet another embodiment, the solanezumab is delivered at a concentration of about 200 μg. In yet another embodiment, the compound is BnH12003.

[0172] In one embodiment, the compound of the present invention is selected for its ability to reduce the expression of HDAC4 in the rodent barrel cortex (layer 4). In one embodiment, the compound is BnH12003.

[0173] In one embodiment, the compound of the present invention is selected for its ability to reduce symptoms of post-traumatic stress disorder.In one embodiment, said selection is carried out by measuring the freezing percentage of a rodent model of post-traumatic stress disorder, in which rodent is conditioned to freeze in response to environmental signals.In one embodiment, said compound is BnH12003.

[0174] Detailed examples of methods useful for selecting compounds of the present invention are described herein.

[0175] V. Therapeutic Uses of Compounds

[0176] The compounds of the present invention can be useful for inhibiting at least one HDAC type.In one embodiment, the compounds described herein can be therapeutically useful, because certain HDACs are known to be involved in the biological pathway that governs higher brain function.Therefore, the compounds described herein can be useful when brain pathology is associated with or mediated by HDAC.

[0177] Recent reports have detailed the importance of histone acetylation in central nervous system (CNS) functions such as neuronal differentiation, memory formation, drug addiction, and depression (Citrome, Psychopharmacol. Bull. 2003, 37, Suppl. 2, 74-88; Johannessen, CNS Drug Rev. 2003, 9, 199-216; Tsankova et al, 2006, Nat. Neurosci. 9, 519-525).

[0178] A neurological disorder is a disease that has as its component a dysfunction of the central or peripheral nervous system. A neurological disorder may result from developmental abnormalities, disease, genetic defects, injury, or toxins that cause a disturbance in the structure or function of the nervous system. Such disorders may affect the central nervous system (e.g., the brain, brain stem, and cerebellum), the peripheral nervous system (e.g., the cranial nerves, spinal nerves, and the sympathetic and parasympathetic nervous systems), and / or the autonomic nervous system (e.g., the part of the nervous system that regulates involuntary actions and is divided into the sympathetic and parasympathetic nervous systems).

[0179] Thus, provided herein is a method for treating or preventing the neurological or neurodegenerative disorder mediated by HDAC in a subject in need of treatment or prevention, comprising administering to the subject a therapeutically effective amount of at least one compound as described herein.In some embodiments, the compound as described herein can be useful for regulating the activity of class IIa HDAC.In some embodiments, the compound is an inhibitor of HDAC4.

[0180] Most neurodegenerative disorders are caused by the degeneration of neurons in the brain. Neurons containing HDAC4 are widely distributed in the brain (Mielcarek et al., 2013, PLoS Biol. 11:e1001717; Takase et al., 2013, PloS ONE 8:e58473). Thus, a treatment such as the present invention that modulates the activity of HDAC4 in the brain can be useful for any neurodegenerative disease of the brain, because the mechanism of intervention contemplated by the present invention can target HDAC4 in neurons regardless of the observed brain pathology. There is no neurodegenerative disease for which the compounds of the present invention are not useful. The compounds of the present invention are not bound by the current understanding of the molecular mechanisms that involve HDAC4 in neurons of the brain. Thus, the compounds of the present invention can be useful for any chronic or acute neurodegenerative disorder, as long as the neurons affected by the pathological manifestations of the disease express HDAC4.

[0181] Examples of chronic neurodegenerative disorders include: chronic neurodegenerative diseases, such as familial and sporadic amyotrophic lateral sclerosis (FALS and ALS, respectively), familial and sporadic Parkinson's disease, Huntington's disease, familial and sporadic Alzheimer's disease, multiple sclerosis, muscular dystrophies, olivopontocerebellar atrophy, multiple system atrophy, Wilson's disease, progressive supranuclear palsy, diffuse Lewy body disease, corticodentatonigral degeneration, progressive familial myoclonic epilepsy, striatonigral degeneration, torsion dystonia, familial tremor, Down syndrome, Tourette's syndrome, Hallervorden-Spatz disease, diabetic peripheral neuropathy, dementia pugilistica, AIDS dementia, age-related dementia, age-related memory impairment, and amyloidosis-related neurodegenerative diseases, such as diseases induced by prion protein (PrP) associated with transmissible spongiform encephalopathies (Creutzfeldt-Jakob disease, Gerstmann-Sträussler-Scheinker syndrome, scrapie, and kuru), and diseases due to excess accumulation of cystatin C (hereditary cystatin C vasculopathy). Examples of acute neurodegenerative disorders include: traumatic brain injury (e.g., surgery-related brain injury), cerebral edema, peripheral nerve injury, spinal cord injury, Leigh's disease, Guillain-Barré syndrome, lysosomal storage disorders such as lipofuscinosis, Alper's disease, restless legs syndrome, dizziness due to CNS degeneration, etc.; pathology occurring with chronic alcohol or drug abuse, including, for example, degeneration of neurons in the locus coeruleus and cerebellum, drug-induced movement disorders; pathology occurring with aging, including degeneration of cerebellar and cortical neurons leading to cognitive and motor disorders; pathology occurring with chronic amphetamine abuse, including degeneration of basal ganglia neurons leading to movement disorders; pathological changes due to local trauma, such as stroke, focal ischemia, vascular insufficiency, hypoxic-ischemic encephalopathy, hyperglycemia, hypoglycemia, or direct trauma; pathology occurring with negative side effects of therapeutic drugs and therapies (e.g., NMD of glutamate receptors). Degeneration of cingulate and entorhinal cortex neurons in response to doses of A-class antagonist anticonvulsants) and Bernike-Korsakoff-related dementia.

[0182] In addition to acute or chronic degenerative disorders, the compounds of the present invention can be useful for treating other types of brain diseases, as long as the affected neurons are partially regulated by HDAC4.Examples of such diseases include: neuropsychiatric disorders associated with cognition, dementia, social behavior, anxiety, or other personality disorders, such as schizophrenia, delirium, attention deficit disorder (ADD), bipolar disorder, obsessive-compulsive disorder, or eating disorder; fear and anxiety disorders, such as panic disorder, various phobias, post-traumatic stress disorder (PTSD), psychogenic erectile dysfunction, or insomnia; various forms of addiction; and mood disorders, such as depression.

[0183] The types of diseases that can benefit from the use of the compounds described herein are not limited to brain diseases. Any neurological disorder in which affected neurons are regulated by HDAC4 can potentially benefit from the compounds of the present invention. As described herein, Alzheimer's disease and PTSD, although the known causes of pathology and disease are very different, benefit from the treatment of the compounds described herein. In the case of PTSD, BnH12003 caused such rodents to release from trauma at a much faster rate than fear-conditioned rodents that were not treated with it. Examples of neurological disorders that can benefit from the compounds of the present invention include: peripheral nervous system diseases due to degeneration or damage, such as ataxia or trauma; and acute or chronic degenerative diseases of the eye, such as glaucoma.

[0184] Insofar as the plasticity of affected neurons is partially regulated by HDAC4, memory loss or cognitive decline can also benefit from treatment using the compounds of the present invention. Examples of such disorders include: memory loss / impairment, disorders associated with cognitive functions such as creativity, problem solving, and intuition.

[0185] Memory includes short-term memory (also called working or recent memory) and long-term memory. Short-term memory includes recent events, while long-term memory is associated with recalling events from the more distant past. Methods for assessing memory recall ability are known to those skilled in the art and can include routine cognitive tests.

[0186] Age-related memory loss is characterized by objective memory loss at an older age compared to when the subject was younger, but cognitive test performance is within the normal range for the subject's age. Subjects with age-related memory loss score within the normal range on standardized diagnostic tests for dementia, as described in DSM-IV. In addition, DSM-IV provides separate diagnostic criteria for a disorder called age-related cognitive decline. Age-related memory loss can include loss of brain weight, gyral atrophy, ventricular enlargement, and selective loss of neurons in various brain regions. For some embodiments of the present invention, more progressive forms of memory loss are also included in the definition of age-related memory disorder. Thus, individuals who have memory loss and cognitive impairment above normal age but score below the diagnostic threshold for Frank dementia may be said to have mild neurocognitive impairment, mild cognitive impairment, senile amnesia, benign senile amnesia, early dementia, provisional dementia, and the like. Such subjects may be somewhat more likely to develop Frank Dementia in later life (see also US Patent Application No. 2006 / 008517).

[0187] Symptoms associated with age-related memory loss include, but are not limited to, alterations in biochemical markers associated with aging brain, such as IL-1β, IFN-γ, p-JNK, p-ERK, and decreased synaptic activity or function, such as synaptic plasticity, as evidenced by decreased long-term potentiation, decreased memory and learning.In some embodiments, the compounds of the present invention modulate the activity of IL-1β, IFN-γ, p-JNK, or p-ERK.

[0188] The method for improving memory can include memory access resetting and memory recovery. The term access resetting as used herein refers to increasing memory exploration. Without being bound by the mechanism of action, it is believed that the compounds of the present invention are effective in increasing memory exploration by resetting synaptic networks. The process of resetting synaptic networks can include increasing the number of active brain synapses and / or reversing neuronal loss.

[0189] In some cases, learning is referred to as memory. Learning is different from memory enhancement, and refers to the ability to generate new memories that did not exist before. In some embodiments, at least one compound of the present invention is administered before or at the same time as memory is generated, in order to test the ability of the compound to affect learning ability rather than the ability of the subject to recall old memories. In some embodiments, at least one compound described herein is administered after memory is generated, preferably after memory is lost, in order to test the ability of the compound to affect the recall of previously generated memories.

[0190] Also provided is a method of improving learning and memory formation in a subject suffering from memory loss, comprising administering to the subject an effective amount of at least one compound described herein, which method may be equally useful for improving learning and memory formation in normal subjects.

[0191] Neurogenesis, or the birth of new nerve cells, was thought to occur only in developing organisms. However, recent studies have demonstrated that neurogenesis continues into and throughout adulthood. Ongoing neurogenesis is believed to be an important mechanism underlying neuroplasticity, allowing organisms to adapt to environmental changes and affecting learning and memory throughout life. In one aspect, the invention includes a method of increasing synaptic density in a subject, comprising administering to a subject in need of increased synaptic density a compound of the invention or a pharma- ceutically acceptable salt, hydrate, solvate, or prodrug thereof. In one aspect, the invention includes a method of increasing synaptic plasticity in a subject, comprising administering to a subject in need of increased synaptic plasticity a compound of the invention or a pharma- ceutically acceptable salt, hydrate, solvate, or prodrug thereof. In one aspect, the invention includes a method of increasing dendritic density of neurons in a subject, comprising administering to a subject in need of increased dendritic density of neurons a compound of the invention or a pharma- ceutically acceptable salt, hydrate, solvate, or prodrug thereof.

[0192] The present invention provides a method for improving memory in subjects with memory impairment. Examples of types of memory impairment include Alzheimer's disease, absent-minded professor, amnesia, memory loss, anticipatory amnesia, blackout (alcohol-related memory loss), Brome's syndrome, childhood amnesia, false memory syndrome, dissociative fugue state, excess memory, Korsakoff's syndrome, partial amnesia, memory distrust syndrome, post-traumatic amnesia, prosopamnesia, psychogenic amnesia, suppressed memory, retrograde amnesia, Ribot's Law, selective amnesia, source amnesia, source monitoring error, seven sins of memory, tip of the tongue, transient epileptic amnesia, transient global amnesia, and twilight sleep.

[0193] In one embodiment, the memory disorder is Alzheimer's disease.Such a method comprises selectively administering an inhibitor and monitoring the subject to confirm the recovery of previously lost memory.The subject can be monitored by routine tests known in the art.

[0194] In another embodiment, the subject with Alzheimer's disease is a subject suffering from late stage Alzheimer's disease. Many compounds proposed for the treatment of Alzheimer's disease are designed to prevent plaque accumulation and treat the early stages of the disease. The compounds of the present invention are useful for treating both early and late stage dementia because they improve memory and cognition rather than just preventing plaque accumulation.

[0195] In one embodiment, the disease or disorder mediated by HDAC is cognitive dysfunction / injury.Therefore, also provided is the method for promoting cognitive function in the subject suffering from cognitive dysfunction / injury, comprising administering to the subject an effective amount of at least one compound as described herein.The present invention also provides the method for promoting cognitive function in normal subjects.

[0196] In some cases, cognitive function is reduced by about 5%, about 10%, about 30%, or more compared to cognitive function measured in age-matched normal subjects. Cognitive function can be promoted to any detectable degree, but in humans, is preferably promoted sufficiently to allow the injured subject to carry out the daily activities of normal life.

[0197] Cognitive function can be assessed, and optionally defined, by one or more tests or assays of cognitive function. Non-limiting examples of tests or assays relating to cognitive function include the CANTAB (see, e.g., Fray et al. "CANTAB battery: proposed utility in neurotoxicology." Neurotoxicol Teratol 1996; 18(4):499-504), the Stroop Test, Trail Making, Wechsler Digit Span, or the CogState computerized cognitive test (see, e.g., Dehaene et al. "Reward-dependent learning in neuronal networks for planning and decision making." Brain Res. Brain Res. 2000; 126:21729; Iverson et al. "Interpreting change on the WAIS-III / WMS-I11 in clinical samples." Arch Clin Neuropsychol. 2001; 16(2):183-91; and Weaver et al. "Mild memory impairment in healthy older adults is distinct from normal aging." Cogn. 2006; 60(2):146-55). The methods of the present invention can be used to promote cognitive function in a normal subject, treat / treat, alleviate / alleviate a cognitively impaired subject, or prevent cognitive impairment in a subject. A normal subject as used herein is a subject who has not been diagnosed with a disorder associated with cognitive impairment.

[0198] In some embodiments, the cognitive dysfunction or damage is associated with, but is not limited to, Alzheimer's disease, Huntington's disease, seizure-induced memory loss, schizophrenia, Rubinstein-Taybi syndrome, Rett syndrome, fragile X syndrome, dementia with Lewy bodies, vascular dementia, bipolar disorder, and social, cognitive and learning disorders associated with autism spectrum disorder (ASD), attention deficit hyperactivity disorder (ADHD), dyslexia, learning disabilities, traumatic head injury, stroke-induced cognitive and motor disorders, traumatic brain injury, neurodegeneration and neuronal loss-mediated cognitive disorders, and attention deficit disorder.

[0199] In some embodiments, the cognitive impairment or damage is associated with, but is not limited to, anxiety disorder, conditioned fear response, panic disorder, obsessive-compulsive disorder, post-traumatic stress disorder, phobia, social anxiety disorder, substance abuse recovery or age-associated memory impairment (AAMI), and age-associated cognitive decline (ARCD).

[0200] Administration of HDAC inhibitors sodium butyrate or trichostatin A has been demonstrated to promote fear extinction in mice, and such promotion is reflected by commonly used behavioral manipulations and is consistent with other studies showing the role of the hippocampus in the extinction of contextual fear (Lattal, et al., 2007, Behav. Neurosci. 121, 5, 1125-1131). In one embodiment, the disease or disorder mediated by HDAC is extinction learning disorder, such as fear extinction deficiency. Thus, also provided is a method for treating or preventing extinction learning disorder in a subject, comprising administering to a subject suffering from extinction learning disorder an effective amount of at least one compound described herein.

[0201] The compounds of the present invention can be used to promote the psychological process of extinction learning, and are therefore useful for treating, alleviating, and / or preventing neuropsychiatric and other related disorders.Unlike traditional anxiolytics that are administered for a long period of time and treat the physiological symptoms of anxiety, the compounds of the present invention can be used for long or short periods of time in conjunction with a second therapy, such as psychotherapy.

[0202] Although not thoroughly investigated herein, HDAC4 mediates gene expression in other cell types, including immune cells.The compounds of the present invention are not bound by the current understanding of the molecular mechanism that involves HDAC4 in immune cells or other types of cells found in cancer.Thus, the compounds of the present invention can be useful for any disease, including hyperproliferative disease, such as malignant tumors of various origins; immune system disease, such as autoimmune disease, inflammation or allergy; infectious disease; transplant rejection; and fibrotic disease.

[0203] In some embodiments, the disease or disorder mediated by HDAC is an immune disorder that can be co-treated with TNFα or other immunomodulatory agents when a therapeutically effective amount of at least one compound as described herein is administered to the subject.Therefore, a method of treating an immune disorder mediated by HDAC in a subject is also provided, comprising administering a therapeutically effective amount of at least one compound as described herein to a subject in need of treatment of the immune disorder before, during, or after treatment with TNFα or other immunomodulatory agents.

[0204] In some embodiments, the HDAC-mediated disease or disorder includes disorders that can also be treated by progenitor / stem cell-based therapy, such as: diabetes-related disorders (organ failure, cirrhosis, and hepatitis); central nervous system (CNS) disorders associated with dysregulation of progenitor cells in the brain (e.g., PTSD); tumors (e.g., retinoblastoma); disorders affecting oligodendrocyte progenitor cells (e.g., astrocytoma and mesentery cell tumors); multiple sclerosis; demyelinating disorders such as leukodystrophies; neurological disorders associated with white matter loss; cerebellar disorders such as ataxia; and olfactory progenitor cell disorders (e.g., anosmia). Thus, methods of treating a HDAC-mediated disorder in a subject are also provided, comprising administering a therapeutically effective amount of at least one compound described herein to a subject in need of treatment for a HDAC-mediated disorder before, during, or after treatment with a progenitor / stem cell-based therapy.

[0205] In some embodiments, the disease or disorder mediated by HDAC includes blood pressure disorders associated with nitric oxide (NO) regulation (e.g., hypertension, erectile dysfunction, asthma; and ocular disorders such as glaucoma).Therefore, there is also provided a method for treating blood pressure disorders associated with HDAC-mediated nitric oxide (NO) regulation in a subject, comprising administering to the subject in need of treatment of a blood pressure disorder associated with HDAC-mediated nitric oxide (NO) regulation a therapeutically effective amount of at least one compound described herein.

[0206] In some embodiments, the disease or disorder is a cardiac hypertrophy disorder.Therefore, also provided is a method for treating a cardiac hypertrophy disorder mediated by HDAC in a subject, comprising administering a therapeutically effective amount of at least one compound described herein to the subject in need of such treatment.

[0207] In some embodiments, the disease or disorder mediated by HDAC includes hematological disorders, such as thalassemia, anemia, and sickle cell anemia. Accordingly, there is also provided a method of treating a HDAC-mediated blood disorder in a subject, comprising administering to the subject in need of treatment thereof a therapeutically effective amount of at least one compound described herein.

[0208] In some embodiments, the disease or disorder mediated by HDAC comprises a metabolic disease, such as prediabetes or diabetes (type 1 or type 2). Accordingly, methods of treating a metabolic disease in a subject, including administering to a subject in need of treatment for a metabolic disease, such as prediabetes or diabetes (type 1 or type 2) mediated by HDAC, a therapeutically effective amount of at least one compound described herein is also provided.

[0209] In one aspect, the present invention provides pharmaceutical compositions further comprising a second therapeutic agent for treating HDAC-mediated disease or disorder in subjects suffering from HDAC-mediated disease or disorder.In some embodiments, the second therapeutic agent is formulated to treat HDAC, preferably HDAC4-mediated disease or disorder.

[0210] Also provided are methods for treating an HDAC-mediated disease or disorder in a subject suffering from an HDAC-mediated disease or disorder, wherein at least one compound described herein, or a pharma- ceutically acceptable salt thereof, is administered to the subject together with one or more additional active agents.

[0211] In some embodiments, the second therapeutic agent is selected from the group consisting of donepezil, rivastigmine, galantamine, memantine, verubecestat, solanezumab, bapineuzumab, aducanumab, lecanemab, tideglusib, epothilone D, and ABBV-8E12.

[0212] In some embodiments, the second therapeutic agent is selected from the group consisting of a cholinesterase inhibitor, an NMDA receptor antagonist, a humanized antibody targeting tau protein, a humanized antibody targeting amyloid beta protein, and a BACE inhibitor.

[0213] Typical therapeutic methods included in the present invention include combination therapy. For example, combination therapy may be a drug therapy (i.e., a compound of the present invention) and a behavioral therapy. Behavioral therapy includes, but is not limited to, electroconvulsive seizure therapy, exercise, group therapy, conversation therapy, or conditioning. In yet another embodiment, the behavioral therapy is cognitive behavioral therapy. Examples of behavioral therapies that can be used in the ongoing method are described, for example, in Cognitive-Behavioral Therapies by K. Dobson, ed., Guilford Publications, Inc., 2002 and The new Handbook of Cognitive Therapy: Basics and Beyond by Judith SS Beck, Guilford Publications, Inc. 1995, the entire contents of which are incorporated herein by reference. Any pharmacologic active ingredient recognized by those skilled in the art as a pharmacological agent that enhances learning or conditioning can be used in the method of the present invention. For example, one class of pharmacologic active ingredients considered herein includes compounds that increase the level of norepinephrine in the brain. Such compounds include compounds that act as norepinephrine reuptake inhibitors, such as tomoxetine, reboxetine, duloxetine, venlafaxine, and milnacipran, and compounds that induce the release of norepinephrine, such as amphetamine, dextroamphetamine, pemoline, and methylphenidate.Another class of such pharmacoactive ingredients is compounds that increase the level of acetylcholine in the brain, including, for example, compounds that block the breakdown of acetylcholine.Examples of such compounds include donepezil HCl or Aricept, which inhibit cholinesterase activity. TM and tacrine.

[0214] The method for combination therapy can include subjecting a subject to one or more sessions of a combination therapy protocol, where the combination therapy protocol includes acute administration of a therapeutically effective amount of the compound of the present invention that enhances learning or conditioning together with a psychotherapy session. In one embodiment, exposure to the compound occurs within about 24 hours, preferably within about 12 hours, more preferably within about 6 hours before starting the psychotherapy session. The entire course of treatment of a neuropsychiatric disorder involves at least one session of such a combination therapy protocol.

[0215] The present invention will now be described in more detail with reference to the following examples, which are presented to aid in the understanding of the present invention and are not intended, and should not be construed as, limiting its scope in any manner.

[0216] Methods for obtaining the compounds described herein, or pharma- ceutically acceptable salts thereof, will be apparent to those skilled in the art, and suitable procedures are described, for example, in the preparations below and in the references cited therein.

[0217] VI. Methods for Producing the Active Compounds

[0218] Methods for synthesizing the compounds of the present invention are provided herein. The names and structures of the compounds are presented in Tables 2 and 3.

[0219] [Table 2]

[0220] JPEG2025512899000010.jpg209146

[0221] JPEG2025512899000011.jpg218146

[0222] JPEG2025512899000012.jpg219146

[0223] JPEG2025512899000013.jpg45146

[0224]

[0225] [Table 3]

[0226] JPEG2025512899000015.jpg218146

[0227] JPEG2025512899000016.jpg220146

[0228] JPEG2025512899000017.jpg220146

[0229] JPEG2025512899000018.jpg164146

[0230]

[0231] Methods for obtaining the compounds of the invention described herein, their pharma- ceutically acceptable salts, or their stereoisomers will be apparent to those of ordinary skill in the art, and suitable methods are described, by way of illustration, in the Preparations and Examples below, and in the references cited in this application.

[0232] Manufacturing Example

[0233] Preparation Example 1. Preparation of active compounds according to representative reaction scheme 1

[0234] [Reaction Scheme 1]

[0235] JPEG2025512899000019.jpg32143

[0236] JPEG2025512899000020.jpg106125

[0237] (1) Synthesis of compound BnH12036

[0238] (3R,6S)-6-Methyl-1-(2-(quinoxalin-6-yl)acetyl)piperidine-3-carboxylic acid

[0239] Step 1: Synthesis of methyl (3R,6S)-6-methyl-1-(2-(quinoxalin-6-yl)acetyl)piperidine-3-carboxylate

[0240] To a solution of 2-(quinoxalin-6-yl)acetic acid (100 mg, 0.531 mmol) in DMF (2.50 mL) was added methyl (3R,6S)-rel-6-methyl-3-piperidinecarboxylate (87.7 mg, 0.558 mg), DIPEA (0.139 mL, 0.797 mmol), HOBt (86.2 mg, 0.638 mmol), and EDC·HCl (99.0 mg, 0.638 mmol). The mixture was stirred at room temperature overnight. The mixture was diluted with EtOAc (5.00 mL) and washed with saturated aqueous NH4Cl (5.00 mL) and brine (5.00 mL). The organic layer was dried over MgSO4, filtered, and concentrated under reduced pressure. The residue was purified by column chromatography on SiO2 (DCM:MeOH=0-5%) to give the desired compound (80.4 mg, 46%) as a dark yellow oil.

[0241] Step 2: Synthesis of compound BnH12036

[0242] To a solution of methyl (3R,6S)-6-methyl-1-(2-(quinoxalin-6-yl)acetyl)piperidine-3-carboxylate (72.5 mg, 0.221 mmol) in THF (1.50 mL) was added 1N NaOH (0.332 mL, 0.332 mmol). The mixture was stirred at room temperature for 2 h. The mixture was diluted with H2O (5.00 mL) and washed twice with DCM (5.00 mL x 2). The aqueous layer was acidified with 1N HCl at 0 °C until pH 2-3 and extracted with EtOAc (5.00 mL). The organic layer was washed with brine (5.00 mL), dried over MgSO4, filtered, and concentrated under reduced pressure to provide compound BnH12036 (20.0 mg, 28%) as a white solid.

[0243] 1H NMR (400 MHz, DMSO-d6) δ 12.42 (s, 1H), 8.92 (d, J = 4.1 Hz, 2H), 8.04 (d, J = 8.6 Hz, 1H), 7.94 (d, J = 23.0 Hz, 1H), 7.73 (dd, J = 14.3, 8.6 Hz, 1H), 4.76 (d, J = 8.4 Hz, 0.5H), 4.55 (dd, J = 13.5, 4.2 Hz, 0.5H), 4.36 (d, J = 8.9 Hz, 0.5H), 1.44 - 3.97 (m, 2.5H), 3.08 (t, J = 12.8 Hz, 0.5H), 2.65 (t, J = 12.6 Hz, 0.5H), 2.29 - 2.17 (m, 1H), 1.83 - 1.78 (m, 1H), 1.73 - 1.62 (m, 1H), 1.56 - 1.54 (m, 2H), 1.10 (dd, J = 18.1, 6.9Hz, 3H).

[0244] (2) Synthesis of compound BnH12037

[0245] (3R,6S)-1-(2-(2,3-dihydrobenzo[b][1,4]dioxin-6-yl)acetyl)-6-methylpiperidine-3-carboxylic acid

[0246] Step 1: Synthesis of methyl (3R,6S)-1-(2-(2,3-dihydrobenzo[b][1,4]dioxin-6-yl)acetyl)-6-methylpiperidine-3-carboxylate

[0247] To a solution of 2-(2,3-dihydrobenzo[b][1,4]dioxin-6-yl)acetic acid (100 mg, 0.515 mmol) in DMF (2.50 mL) was added methyl (3R,6S)-rel-6-methyl-3-piperidinecarboxylate (85.0 mg, 0.541 mmol), DIPEA (0.135 mL, 0.772 mmol), HOBt (83.5 mg, 0.618 mmol), and EDC·HCl (95.9 mg, 0.618 mmol). The mixture was stirred at room temperature overnight. The mixture was diluted with EtOAc (5.00 mL) and washed with saturated aqueous NH4Cl (5.00 mL) and brine (5.00 mL). The organic layer was dried over MgSO4, filtered, and concentrated under reduced pressure. The residue was purified by column chromatography on SiO2 (DCM:MeOH=0-5%) to give the desired compound (62.0 mg, 38%) as a dark yellow oil.

[0248] Step 2: Synthesis of compound BnH12037

[0249] To a solution of methyl (3R,6S)-1-(2-(2,3-dihydrobenzo[b][1,4]dioxin-6-yl)acetyl)-6-methylpiperidine-3-carboxylate (135 mg, 0.405 mmol) in THF (2.00 mL) was added 1N NaOH (0.607 mL, 0.607 mmol). The mixture was stirred at room temperature for 2 h. The mixture was diluted with H2O (5.00 mL) and washed twice with DCM (5.00 mL x 2). The aqueous layer was acidified with 1N HCl at 0 °C until pH 1-2 and extracted with EtOAc (5.00 mL). The organic layer was washed with brine (5.00 mL), dried over MgSO4, filtered, and concentrated under reduced pressure to provide compound BnH12037 (90.0 mg, 69%) as a white solid.

[0250] 1H NMR (400 MHz, DMSO-d6) δ 12.36 (s, 1H), 6.78 - 6.61 (m, 3H), 4.71 (s, 0.5H), 4.51 (dd, J = 13.3, 4.3 Hz, 0.5H), 4.20 (s, 4H), 3.88 - 3.84 (m, 0.5H), 3.63 - 3.54 (m, 2H), 2.97 (t, J = 12.9 Hz, 0.5H), 2.58 (t, J = 12.7 Hz, 0.5H), 2.18 - 2.09 (m, 1H), 1.81 - 1.75 (m, 1H), 1.69 - 1.59 (m, 1H), 1.50 - 1.43 (m, 2H), 1.03 (dd, J = 7.0, 3.4 Hz, 3H).

[0251] (3) Synthesis of compound BnH12042

[0252] (3R,6S)-1-(2-([1,1'-biphenyl]-4-yl)acetyl)-6-methylpiperidine-3-carboxylic acid

[0253] Step 1: Synthesis of methyl (3R,6S)-1-(2-([1,1'-biphenyl]-4-yl)acetyl)-6-methylpiperidine-3-carboxylate

[0254] To a solution of 4-biphenylacetic acid (110 mg, 0.518 mmol) in DMF (1.73 mL) was added methyl (3R,6S)-rel-6-methyl-3-piperidinecarboxylate (85.6 mg, 0.544 mmol), DIPEA (0.135 mL, 0.777 mmol), HOBt (84.0 mg, 0.622 mmol), and EDC·HCl (96.6 mg, 0.622 mmol). The mixture was stirred at room temperature for 17 h. The mixture was diluted with EtOAc (5.00 mL) and washed with saturated aqueous NH4Cl (5.00 mL) and brine (5.00 mL). The organic layer was dried over MgSO4, filtered, and concentrated under reduced pressure. The residue was purified by column chromatography on SiO2 (n-hexane:EtOAc = 3:1) to give the desired compound (145 mg, 79%) as a colorless oil.

[0255] Step 2: Synthesis of compound BnH12042

[0256] To a solution of methyl (3R,6S)-1-(2-([1,1'-biphenyl]-4-yl)acetyl)-6-methylpiperidine-3-carboxylate (145 mg, 0.413 mmol) in THF (1.42 mL) was added 1N NaOH (0.619 mL, 0.619 mmol). The mixture was stirred at room temperature for 2 h. The mixture was diluted with H2O (3.00 mL) and washed twice with DCM (3.00 mL x 2). The aqueous layer was acidified with 1N HCl at 0 °C until pH 1-2 and extracted with EtOAc (3.00 mL). The organic layer was washed with brine (3.00 mL), dried over MgSO4, filtered and concentrated under reduced pressure to provide compound BnH12042 (120 mg, 86%) as a white solid.

[0257] 1H NMR (400 MHz, Chloroform-d) δ 7.57 (dd, J = 10.6, 7.8 Hz, 4H), 7.43 (t, J = 7.5 Hz, 2H), 7.33 (dd, J = 10.8, 7.6 Hz, 3H), 4.99 (s, 0.5H), 4.84 (d, J = 13.7 Hz, 0.5H), 4.22 (s, 0.5H), 3.96 (d, J = 13.8 Hz, 0.5H), 3.79 (s, 2H), 3.17 (t, J = 12.9 Hz, 0.5H), 2.79 (t, J = 12.9 Hz, 0.5H), 2.41 (s, 0.5H), 2.14 (s, 0.5H), 1.92 (d, J = 13.0 Hz, 1H), 1.80 (d, J = 13.6 Hz, 1H), 1.58 (d, J = 24.0 Hz, 2H), 1.15 (d, J = 6.8 Hz, 3H).

[0258] (4) Synthesis of compound BnH12078

[0259] (3R,6S)-1-(2-([1,1'-biphenyl]-3-yl)acetyl)-6-methylpiperidine-3-carboxylic acid

[0260] Step 1: Synthesis of methyl (3R,6S)-1-(2-([1,1'-biphenyl]-3-yl)acetyl)-6-methylpiperidine-3-carboxylate

[0261] To a solution of 3-biphenylylacetic acid (70.0 mg, 0.328 mmol) in DMF (1.09 mL) was added methyl (3R,6S)-rel-6-methyl-3-piperidinecarboxylate (54.2 mg, 0.345 mmol), DIPEA (0.086 mL, 0.492 mmol), HOBt (53.2 mg, 0.394 mmol), and EDC·HCl (61.2 mg, 0.394 mmol). The mixture was stirred at room temperature overnight. The mixture was diluted with EtOAc (5.00 mL) and washed with saturated aqueous NH4Cl (5.00 mL) and brine (5.00 mL). The organic layer was dried over MgSO4, filtered, and concentrated under reduced pressure. The residue was purified by column chromatography on SiO2 (n-hexane:EtOAc = 2:1) to give the desired compound (69.2 mg, 60%) as a colorless oil.

[0262] Step 2: Synthesis of compound BnH12078

[0263] To a solution of (3R,6S)-1-(2-([1,1'-biphenyl]-3-yl)acetyl)-6-methylpiperidine-3-carboxylate (69.2 mg, 0.196 mmol) in THF (0.675 mL) was added 1N NaOH (0.294 mL, 0.294 mmol). The mixture was stirred at room temperature for 2 h. The mixture was diluted with H2O (3.00 mL) and washed twice with EtOAc (3.00 mL x 2). The aqueous layer was acidified with 1N HCl at 0 °C until pH 1-2 and extracted with EtOAc (3.00 mL). The organic layer was washed with brine (3.00 mL), dried over MgSO4, filtered, and concentrated under reduced pressure. The residue was purified by column chromatography on SiO2 (10% MeOH in DCM) to give compound BnH12078 (50.0 mg, 76%) as a white solid.

[0264] 1H NMR (400 MHz, DMSO-d6) δ 12.43 (s, 1H), 7.63 (t, J = 6.1 Hz, 2H), 7.49 (dt, J = 15.4, 9.1 Hz, 4H), 7.39 (dt, J = 14.3, 7.5 Hz, 2H), 7.22 (dd, J = 16.7, 7.6 Hz, 1H), 4.75 (s, 0.5H), 4.54 (d, J = 13.6 Hz, 0.5H), 4.32 (s, 0.5H), 3.96 (d, J = 13.8 Hz, 0.5H), 3.89 - 3.72 (m, 2H), 3.03 (t, J = 12.9 Hz, 0.5H), 2.62 (t, J = 12.7 Hz, 0.5H), 2.16 (d, J = 12.8 Hz, 1H), 1.79 (d, J = 12.8 Hz, 1H), 1.71 - 1.59 (m, 1H), 1.57 - 1.42 (m, 2H), 1.06 (t, J = 6.5 Hz, 3H).

[0265] (5) Synthesis of compound BnH12080

[0266] (3R,6S)-6-Methyl-1-(4-(pyridin-3-yl)benzoyl)piperidine-3-carboxylic acid

[0267] Step 1: Synthesis of methyl (3R,6S)-6-methyl-1-(4-(pyridin-3-yl)benzoyl)piperidine-3-carboxylate

[0268] To a suspension of 4-(pyridin-3-yl)benzoic acid (100 mg, 0.502 mmol) in DMF (1.67 mL) was added methyl (3R,6S)-6-methylpiperidine-3-carboxylate (82.9 mg, 0.527 mmol), DIPEA (0.132 mL, 0.753 mmol), HOBt (81.4 mg, 0.602 mmol), and EDC·HCl (93.5 mg, 0.602 mmol). The mixture was stirred at room temperature for 2 h. The mixture was diluted with EtOAc (4.00 mL) and washed with saturated aqueous NH4Cl (4.00 mL) and brine (4.00 mL). The organic layer was dried over MgSO4, filtered, and concentrated under reduced pressure. The residue was purified by column chromatography on SiO2 (DCM:MeOH=0-2%) to give the desired compound (82.0 mg, 48.3%) as a pale yellow oil.

[0269] Step 2: Synthesis of compound BnH12080

[0270] To a solution of methyl (3R,6S)-6-methyl-1-(4-(pyridin-3-yl)benzoyl)piperidine-3-carboxylate (82.0 mg, 0.242 mmol) in THF (0.836 mL) was added 1N NaOH (0.363 mL, 0.363 mmol). The mixture was stirred at room temperature for 2 h. The mixture was diluted with H2O (5.00 mL) and washed twice with DCM (5.00 mL x 2). The aqueous layer was acidified with 1N HCl at 0 °C until pH 3-4 and extracted with EtOAc (5.00 mL). The organic layer was washed with brine (5.00 mL), dried over MgSO4, filtered, and concentrated under reduced pressure to provide compound BnH12080 (47.0 mg, 59.8%) as a white solid.

[0271] 1H NMR (400 MHz, DMSO-d6) δ 12.47 (s, 1H), 8.95 (d, J = 2.4 Hz, 1H), 8.62-8.60 (dd, J = 4.8, 1.6 Hz, 1H), 8.15-8.12 (m, 1H), 7.83-7.81 (d, J = 8.0 Hz, 2H), 7.53-7.50 (dd, J = 7.9, 4.5 Hz, 3H), 2.45-2.34 (m, 2H), 1.92-1.88 (d, J = 15.5 Hz, 1.5H), 1.80-1.69 (dt, J = 20.7, 7.5 Hz, 2H), 1.57 (s, 1.5H), 1.21-1.18 (dd, J = 7.1, 2.8 Hz, 4H).

[0272] Preparation Example 2. Preparation of active compound according to representative reaction scheme 3

[0273] [Reaction Scheme 3]

[0274] JPEG2025512899000021.jpg61142

[0275] JPEG2025512899000022.jpg135133

[0276] JPEG2025512899000023.jpg188131

[0277] Step 1: Synthesis of methyl (3R,6S)-6-methyl-1-(2-(4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenyl)acetyl)piperidine-3-carboxylate (compound 3-1)

[0278] The first step of reaction scheme 3 was carried out to obtain compound BnH12032. Then, a suspension of compound BnH12032 (1.60 g, 4.52 mmol), bis(pinacolato)diboron (1.62 g, 4.97 mmol), and KOAc (1.33 g, 13.6 mmol) in 1,4-dioxane (15.6 mL) was vigorously stirred and bubbled with N2 gas for 5 minutes, followed by addition of Pd(dppf)Cl2 (327 mg, 0.447 mmol). The mixture was bubbled with N2 gas for 5 minutes and refluxed at 100 °C for 17 hours. After completion of the reaction, the reaction mixture was filtered through a Celite pad and washed with DCM (10.0 mL x 3). The combined organic layer was washed with H2O (40.0 mL), dried over MgSO4, filtered, and concentrated under reduced pressure. The residue was purified by SiO2 column chromatography (n-hexane:EtOAc = 1:1) to give compound 3-1 (1.40 g, 77%) as a brown oil.

[0279] (1) Synthesis of compound BnH12043

[0280] (3R,6S)-6-Methyl-1-(2-(4-(pyridin-4-yl)phenyl)acetyl)piperidine-3-carboxylic acid

[0281] Step 1: Synthesis of compound BnH12043

[0282] A solution of compound 3-1 (235 mg, 0.586 mmol), 4-bromopyridine (125 mg, 0.644 mmol), and Na2CO3 (186 mg, 1.76 mmol) in MeCN / H2O (2:1, 7.55 mL) was vigorously stirred, bubbled with N2 for 5 min, and Pd(dppf)Cl2 (42.9 mg, 0.059 mmol) was added. The mixture was bubbled with N2 gas for 5 min and refluxed at 90 °C for 3 h. Subsequently, after the mixture was cooled to room temperature, 1N NaOH (1.17 mL, 1.17 mmol) was added to the mixture and stirred at room temperature for 3 h. The mixture was diluted with H2O (3.00 mL) and washed with DCM (3.00 mL × 3). The pH value was adjusted to pH 4-5 by adding 1N HCl to the aqueous layer, and the mixture was extracted with EtOAc (3.00 mL × 3). The combined organic layers were dried over MgSO4, filtered and concentrated under reduced pressure. The residue was purified by column chromatography on SiO2 (3% MeOH in DCM) to give compound BnH12043 (85.0 mg, 42%) as a white solid.

[0283] 1H NMR (400 MHz, DMSO-d6) δ 8.65 - 8.59 (m, 2H), 7.76 (d, J = 7.9 Hz, 2H), 7.71 (d, J = 5.6 Hz, 2H), 7.37 (dd, J = 18.7, 7.9 Hz, 2H), 4.75 (s, 0.5H), 4.60 - 4.50 (m, 0.5H), 4.27 (d, J = 7.7 Hz, 0.5H), 3.92 (d, J = 13.7 Hz, 0.5H), 3.88 - 3.72 (m, 2H), 3.04 (t, J = 12.9 Hz, 0.5H), 2.63 (d, J = 12.6 Hz, 0.5H), 2.24 - 2.12 (m, 1H), 1.79 (dt, J = 16.9, 5.7 Hz, 1H), 1.65 (d, J = 12.6 Hz, 1H), 1.57 - 1.49 (m, 2H), 1.08 (dt, J = 11.7, 4.9Hz, 3H).

[0284] (2) Synthesis of compound BnH12044

[0285] (3R,6S)-6-Methyl-1-(2-(4-(pyridin-2-yl)phenyl)acetyl)piperidine-3-carboxylic acid

[0286] Step 1: Synthesis of compound BnH12044

[0287] A solution of compound 3-1 (235 mg, 0.586 mmol), 2-bromopyridine (125 mg, 0.644 mmol), and Na2CO3 (186 mg, 1.76 mmol) in MeCN / H2O (2:1, 7.55 mL) was vigorously stirred, bubbled with N2 gas for 5 min, and Pd(dppf)Cl2 (42.9 mg, 0.059 mmol) was added. The mixture was bubbled with N2 gas for 5 min and refluxed at 90 °C for 3 h. Subsequently, the mixture was cooled to room temperature, 1N NaOH (1.17 mL, 1.17 mmol) was added to the mixture, and stirred at room temperature for 3 h. The mixture was diluted with H2O (3.00 mL) and washed with DCM (3.00 mL × 3). The pH value was adjusted to pH 4-5 by adding 1N HCl to the aqueous layer, and the mixture was extracted with EtOAc (3.00 mL × 3). The combined organic layers were dried over MgSO4, filtered and concentrated under reduced pressure. The residue was purified by column chromatography on SiO2 (3% to 5% MeOH in DCM) to give compound BnH12044 (85.0 mg, 42%) as a white solid.

[0288] 1H NMR (400 MHz, Chloroform-d) δ 8.75 (d, J = 23.7 Hz, 1H), 7.89 (dd, J = 26.8, 7.9 Hz, 2H), 7.83 - 7.67 (m, 2H), 7.40 (dd, J = 16.2, 7.9 Hz, 2H), 7.26 (m, 1H), 4.97 (s, 0.5H), 4.81 (d, J = 13.9 Hz, 0.5H), 4.18 (s, 0.5H), 3.99 (d, J = 13.7 Hz, 0.5H), 3.83 (s, 2H), 3.13 (t, J = 12.9 Hz, 0.5H), 2.80 (t, J = 12.9 Hz, 0.5H), 2.40 (s, 0.5H), 2.02 (s, 0.5H), 1.88 (s, 1H), 1.81 - 1.69 (m, 1H), 1.62 - 1.40 (m, 2H), 1.14 (d, J = 6.4 Hz, 3H).

[0289] (3) Synthesis of compound BnH12045

[0290] (3R,6S)-6-Methyl-1-(2-(4-(pyridazin-4-yl)phenyl)acetyl)piperidine-3-carboxylic acid

[0291] Step 1: Synthesis of compound BnH12045

[0292] A solution of compound 3-1 (240 mg, 0.598 mmol), 4-bromopyridazine hydrobromide (158 mg, 0.658 mmol), and Na2CO3 (190 mg, 1.79 mmol) in MeCN / HO (2:1, 7.47 mL) was vigorously stirred, bubbled with N2 gas for 5 min, and Pd(dppf)Cl2 (43.8 mg, 0.060 mmol) was added. The mixture was bubbled with N2 gas for 5 min and refluxed at 90 °C for 3 h. Subsequently, the mixture was cooled to room temperature, 1N NaOH (1.20 mL, 1.20 mmol) was added to the mixture, and stirred at room temperature for 3 h. The mixture was diluted with HO (3.00 mL) and washed with DCM (3.00 mL x 3). The aqueous layer was added with 1N HCl to adjust the pH value to pH 4-5, and the mixture was extracted with EtOAc (3.00 mL×3). The combined organic layers were dried over MgSO4, filtered, and concentrated under reduced pressure. The residue was purified by column chromatography on SiO2 (5% MeOH in DCM) to give compound BnH12045 (65.0 mg, 32%) as a pink solid.

[0293] 1H NMR (400 MHz, Chloroform-d) δ 9.48 (s, 1H), 9.23 (d, J = 5.4 Hz, 1H), 7.65 (d, J = 8.1 Hz, 3H), 7.44 (d, J = 7.9 Hz, 2H), 4.98 (s, 0.5H), 4.84 (d, J = 13.6 Hz, 0.5H), 4.21 (s, 0.5H), 3.94 (d, J = 14.9 Hz, 0.5H), 3.82 (s, 2H), 3.20 (t, J = 12.9 Hz, 0.5H), 2.82 (s, 0.5H), 2.43 (s, 0.5H), 2.20 (s, 0.5H), 1.97 (d, J = 13.7 Hz, 1H), 1.81 (s, 1H), 1.62 (s, 2H), 1.24 - 1.13 (m, 3H).

[0294] (4) Synthesis of compound BnH12046

[0295] (3R,6S)-6-Methyl-1-(2-(4-(pyridin-3-yl)phenyl)acetyl)piperidine-3-carboxylic acid

[0296] Step 1: Synthesis of compound BnH12046

[0297] A solution of compound 3-1 (235 mg, 0.586 mmol), 3-bromopyridine (102 mg, 0.644 mmol), and Na2CO3 (124 mg, 1.17 mmol) in MeCN / H2O (2:1, 7.55 mL) was vigorously stirred, bubbled with N2 gas for 5 min, and Pd(dppf)Cl2 (42.9 mg, 0.059 mmol) was added. The mixture was bubbled with N2 gas for 5 min and refluxed at 90 °C for 3 h. Subsequently, the mixture was cooled to room temperature, 1N NaOH (1.17 mL, 1.17 mmol) was added to the mixture and stirred at room temperature for 3 h. The mixture was diluted with H2O (3.00 mL) and washed with DCM (3.00 mL × 3). The pH value was adjusted to pH 4-5 by adding 1N HCl to the aqueous layer, and the mixture was extracted with EtOAc (3.00 mL × 3). The combined organic layers were dried over MgSO4, filtered and concentrated under reduced pressure. The residue was purified by column chromatography on SiO2 (3% to 5% MeOH in DCM) to give compound BnH12046 (90.2 mg, 45%) as a white solid.

[0298] 1H NMR (400 MHz, DMSO-d6) δ 12.41 (s, 1H), 8.89 (d, J = 2.4 Hz, 1H), 8.56 (dd, J = 4.8, 1.6 Hz, 1H), 8.07 (dt, J = 8.0, 2.0 Hz, 1H), 7.68 (d, J = 7.8 Hz, 2H), 7.48 (dd, J = 7.9, 4.7 Hz, 1H), 7.35 (dd, J = 18.4, 7.8 Hz, 2H), 4.75 (s, 0.5H), 4.57 - 4.49 (m, 0.5H), 4.29 (s, 0.5H), 3.98 - 3.87 (m, 1.80 (dd, J = 13.4, 3.9 Hz, 1H), 1.66 (d, J = 13.6 Hz, 1H), 1.53 (s, 2H), 1.12 - 1.03 (m, 3H).

[0299] (5) Synthesis of compound BnH12047

[0300] (3R,6S)-6-メチル-1-(2-(4-(ピラジン-2-イル)フェニル)アセチル)ピぺリジン-3-カルボン acid

[0301] Stage 1: Synthesis of compound BnH12047

[0302] A solution of compound 3-1 (235 mg, 0.586 mmol), 2-bromopyrazine (102 mg, 0.644 mmol), and Na2CO3 (124 mg, 1.17 mmol) in MeCN / H2O (2:1, 7.55 mL) was vigorously stirred, bubbled with N2 gas for 5 min, and Pd(dppf)Cl2 (42.9 mg, 0.059 mmol) was added. The mixture was bubbled with N2 gas for 5 min and refluxed at 90 °C for 3 h. Subsequently, the mixture was cooled to room temperature, 1N NaOH (1.17 mL, 1.17 mmol) was added to the mixture and stirred at room temperature for 3 h. The mixture was diluted with H2O (3.00 mL) and washed with DCM (3.00 mL × 3). The pH value was adjusted to pH 1-2 by adding 1N HCl to the aqueous layer, and the mixture was extracted with EtOAc (3.00 mL × 3). The combined organic layers were dried over MgSO4, filtered and concentrated under reduced pressure. The residue was purified by column chromatography on SiO2 (3% to 5% MeOH in DCM) to give compound BnH12047 (76.0 mg, 38%) as a white solid.

[0303] 1H NMR (400 MHz, DMSO-d6) δ 12.41 (s, 1H), 9.25 (d, J = 1.5 Hz, 1H), 8.71 (t, J = 2.0 Hz, 1H), 8.60 (d, J = 2.5 Hz, 1H), 8.10 (d, J = 7.9 Hz, 2H), 7.39 (dd, J = 18.2, 7.9 Hz, 2H), 4.75 (s, 0.5H), 4.55 (dd, J = 13.6, 4.2 Hz, 0.5H), 4.28 (s, 0.5H), 3.97 - 3.89 (m, 0.5H), 3.89 - 3.74 (m, 2H), 3.04 (t, J = 12.9 Hz, 0.5H), 2.63 (t, J = 12.7 Hz, 0.5H), 2.19 (tt, J = 12.0, 4.2 Hz, 1H), 1.79 (dd, J = 13.5, 3.9 Hz, 1H), 1.66 (h, J = 13.1, 10.8 Hz, 1H), 1.52 (dq, J = 8.8, 4.9, 4.2 Hz, 2H), 1.07 (dd, J = 9.1, 6.8 Hz, 3H).

[0304] (6) Synthesis of compound BnH12048

[0305] (3R,6S)-6-メチル-1-(2-(4-(ピリミジン-2-イル)フェニル)アセチル)ピぺリジン-3-カルボン acid

[0306] Stage 1: Synthesis of compound BnH12048

[0307] A solution of compound 3-1 (170 mg, 0.424 mmol), 2-bromopyrimidine (67.4 mg, 0.424 mmol), and Na2CO3 (89.8 mg, 0.847 mmol) in MeCN / HO (2:1, 5.47 mL) was vigorously stirred, bubbled with N2 gas for 5 min, and Pd(dppf)Cl2 (31.0 mg, 0.042 mmol) was added. The mixture was bubbled with N2 gas for 5 min and refluxed at 90 °C for 3 h. Subsequently, the mixture was cooled to room temperature, 1N NaOH (0.847 mL, 0.847 mmol) was added to the mixture, and stirred at room temperature for 3 h. The mixture was diluted with HO (2.00 mL) and washed with DCM (3.00 mL x 3). The aqueous layer was added with 1N HCl to adjust the pH value to pH 1-2, and the mixture was extracted with EtOAc (3.00 mL×3). The combined organic layers were dried over MgSO4, filtered, and concentrated under reduced pressure. The residue was purified by column chromatography on SiO2 (3%-5% MeOH in DCM) to give compound BnH12048 (34.0 mg, 23%) as a white solid.

[0308] 1H NMR (400 MHz, Chloroform-d) δ 8.83 (t, J = 5.6 Hz, 2H), 8.37 (dd, J = 16.1, 7.9 Hz, 2H), 7.41 (t, J = 6.8 Hz, 2H), 7.20 (t, J = 4.9 Hz, 1H), 4.98 (s, 0.5H), 4.83 (d, J = 13.7 Hz, 0.5H), 4.17 (s, 0.5H), 3.99 - 3.91 (m, 0.5H), 3.84 (s, 2H), 3.13 (t, J = 13.0 Hz, 0.5H), 2.80 (t, J = 12.8 Hz, 0.5H), 2.40 (s, 0.5H), 2.03 (d, J = 7.0 Hz, 0.5H), 1.88 (s, 1H), 1.76 (t, J = 11.8 Hz, 1H), 1.62 - 1.40 (m, 2H), 1.17 - 1.09 (m, 3H).

[0309] (7) Synthesis of compound BnH12049

[0310] (3R,6S)-1-(2-(4-(5-fluoropyrimidin-2-yl)phenyl)acetyl)-6-methylpiperidine-3-carboxylic acid

[0311] Step 1: Synthesis of compound BnH12049

[0312] A solution of compound 3-1 (170 mg, 0.424 mmol), 2-bromo-5-fluoropyrimidine (82.5 mg, 0.466 mmol), and Na2CO3 (89.8 mg, 0.847 mmol) in MeCN / HO (2:1, 5.47 mL) was vigorously stirred, bubbled with N2 gas for 5 min, and Pd(dppf)Cl2 (31.0 mg, 0.042 mmol) was added. The mixture was bubbled with N2 gas for 5 min and refluxed at 90 °C for 3 h. Subsequently, the mixture was cooled to room temperature, 1N NaOH (0.847 mL, 0.847 mmol) was added to the mixture, and stirred at room temperature for 3 h. The mixture was diluted with HO (2.00 mL) and washed with DCM (3.00 mL x 3). The aqueous layer was added with 1N HCl to adjust the pH value to pH 1-2, and the mixture was extracted with EtOAc (3.00 mL×3). The combined organic layers were dried over MgSO4, filtered, and concentrated under reduced pressure. The residue was purified by column chromatography on SiO2 (3%-5% MeOH in DCM) to give compound BnH12049 (42.0 mg, 27%) as a white solid.

[0313] 1H NMR (400 MHz, Chloroform-d) δ 8.66 (s, 2H), 8.33 (s, 2H), 7.38 (d, J = 8.3 Hz, 2H), 4.98 (s, 0.5H), 4.83 (d, J = 13.8 Hz, 0.5H), 4.18 (s, 0.5H), 3.93 (d, J = 13.9 Hz, 0.5H), 3.82 (s, 2H), 3.14 (t, J = 13.0 Hz, 0.5H), 2.79 (t, J = 12.8 Hz, 0.5H), 2.40 (s, 0.5H), 2.10 (s, 0.5H), 1.91 (d, J = 13.2 Hz, 1H), 1.78 (d, J = 13.3 Hz, 1H), 1.56 (d, J = 31.7 Hz, 2H), 1.13 (s, 3H).

[0314] (8) Synthesis of compound BnH12050

[0315] (3R,6S)-6-Methyl-1-(2-(4-(2-methylpyrimidin-5-yl)phenyl)acetyl)piperidine-3-carboxylic acid

[0316] Step 1: Synthesis of compound BnH12050

[0317] A solution of compound 3-1 (233 mg, 0.581 mmol), 5-bromo-2-methylpyrimidine (110 mg, 0.639 mmol), and Na2CO3 (123 mg, 1.16 mmol) in MeCN / HO (2:1, 7.49 mL) was vigorously stirred, bubbled with N2 gas for 5 min, and Pd(dppf)Cl2 (42.5 mg, 0.058 mmol) was added. The mixture was bubbled with N2 gas for 5 min and refluxed at 90 °C for 3 h. Subsequently, the mixture was cooled to room temperature, 1N NaOH (1.16 mL, 1.16 mmol) was added to the mixture, and stirred at room temperature for 3 h. The mixture was diluted with HO (2.00 mL) and washed with DCM (3.00 mL x 3). The aqueous layer was added with 1N HCl to adjust the pH value to pH 1-2, and the mixture was extracted with EtOAc (3.00 mL×3). The combined organic layers were dried over MgSO4, filtered, and concentrated under reduced pressure. The residue was purified by column chromatography on SiO2 (3%-5% MeOH in DCM) to give compound BnH12050 (45.0 mg, 21%) as a white solid.

[0318] 1H NMR (400 MHz, Chloroform-d) δ 8.88 (d, J = 8.6 Hz, 2H), 7.53 (d, J = 8.0 Hz, 2H), 7.39 (s, 2H), 4.99 (s, 0.5H), 4.84 (d, J = 13.5 Hz, 0.5H), 4.21 (s, 0.5H), 3.92 (d, J = 13.9 Hz, 0.5H), 3.82 (s, 2H), 3.16 (t, J = 12.9 Hz, 0.5H), 2.79 (s, 3.5H), 2.42 (s, 1H), 2.19 (d, J = 11.2 Hz, 0.5H), 1.96 (s, 0.5H), 1.80 (t, J = 13.5 Hz, 1H), 1.61 (d, J = 16.9 Hz, 2H), 1.18 (q, J = 8.0 Hz, 3H).

[0319] (9) Synthesis of compound BnH12051

[0320] (3R,6S)-6-Methyl-1-(2-(4-(2-(trifluoromethyl)pyrimidin-5-yl)phenyl)acetyl)piperidine-3-carboxylic acid

[0321] Step 1: Synthesis of compound BnH12051

[0322] A solution of compound 3-1 (240 mg, 0.598 mmol), 2-chloro-5-(trifluoromethyl)pyrimidine (120 mg, 0.658 mmol), and Na2CO3 (127 mg, 1.20 mmol) in MeCN / HO (2:1, 7.72 mL) was vigorously stirred, bubbled with N2 gas for 5 min, and Pd(dppf)Cl2 (43.8 mg, 0.060 mmol) was added. The mixture was bubbled with N2 gas for 5 min and refluxed at 90 °C for 3 h. Subsequently, the mixture was cooled to room temperature, 1N NaOH (1.20 mL, 1.20 mmol) was added to the mixture, and stirred at room temperature for 3 h. The mixture was diluted with HO (3.00 mL) and washed with DCM (3.00 mL x 3). The aqueous layer was added with 1N HCl to adjust the pH value to pH 1-2, and the mixture was extracted with EtOAc (3.00 mL×3). The combined organic layers were dried over MgSO4, filtered, and concentrated under reduced pressure. The residue was purified by column chromatography on SiO2 (5%-10% MeOH in DCM) to give compound BnH12051 (142 mg, 58%) as a white solid.

[0323] 1H NMR (400 MHz, DMSO-d6) δ 12.44 (s, 1H), 9.33 (s, 2H), 8.40 (dd, J = 8.3, 2.0 Hz, 2H), 7.43 (dd, J = 17.7, 8.0 Hz, 2H), 4.75 (d, J = 9.4 Hz, 0.5H), 4.55 (dd, J = 13.5, 4.1 Hz, 0.5H), 4.26 (d, J = 8.6 Hz, 0.5H), 3.98 - 3.78 (m, 2.5H), 3.04 (t, J = 12.9 Hz, 0.5H), 2.63 (t, J = 12.8Hz, 0.5H), 2.25 - 2.11 (m, 1H), 1.78 (dt, J = 12.4, 3.9 Hz, 1H), 1.66 (dq, J = 19.1, 12.9, 10.3 Hz, 1H), 1.52 (dq, J = 11.8, 7.5, 5.7 Hz, 2H), 1.07 (t, J = 7.2 Hz, 3H).

[0324] (10) Synthesis of compound BnH12052

[0325] (3R,6S)-1-(2-(4-(2-cyclopropylpyrimidin-5-yl)phenyl)acetyl)-6-methylpiperidine-3-carboxylic acid

[0326] Step 1: Synthesis of compound BnH12052

[0327] A solution of compound 3-1 (240 mg, 0.598 mmol), 5-bromo-2-cyclopropylpyrimidine (131 mg, 0.658 mmol), and Na2CO3 (127 mg, 1.20 mmol) in MeCN / HO (2:1, 7.72 mL) was vigorously stirred, bubbled with N2 gas for 5 min, and Pd(dppf)Cl2 (43.8 mg, 0.060 mmol) was added. The mixture was bubbled with N2 gas for 5 min and refluxed at 90 °C for 3 h. Subsequently, the mixture was cooled to room temperature, 1N NaOH (1.20 mL, 1.20 mmol) was added to the mixture, and stirred at room temperature for 3 h. The mixture was diluted with HO (3.00 mL) and washed with DCM (3.00 mL x 3). The aqueous layer was added with 1N HCl to adjust the pH value to pH 1-2, and the mixture was extracted with EtOAc (3.00 mL×3). The combined organic layers were dried over MgSO4, filtered, and concentrated under reduced pressure. The residue was purified by column chromatography on SiO2 (5%-10% MeOH in DCM) to give compound BnH12052 (84.9 mg, 37%) as a white solid.

[0328] 1H NMR (400 MHz, Chloroform-d) δ 8.79 (s, 2H), 7.50 (d, J = 8.0 Hz, 2H), 7.38 (d, J = 8.2 Hz, 2H), 4.99 (s, 0.5H), 4.84 (d, J = 13.8 Hz, 0.5H), 4.21 (s, 0.5H), 3.93 (d, J = 14.0 Hz, 0.5H), 3.80 (s, 2H), 3.17 (t, J = 12.9 Hz, 0.5H), 2.81 (t, J = 12.9 Hz, 0.5H), 2.43 (s, 0.5H), 2.30 (ddd, J = 13.1, 8.2, 4.8 Hz, 1H), 2.20 (s, 0.5H), 1.95 (s, 1H), 1.81 (d, J = 15.7 Hz, 1H), 1.61 (d, J = 16.2 Hz, 2H), 1.15 (dd, J = 16.0, 9.3 Hz, 7H).

[0329] (11) Synthesis of compound BnH12053

[0330] Sodium (3R,6S)-6-methyl-1-(2-(4-(1-methyl-1H-imidazol-2-yl)phenyl)acetyl)piperidine-3-carboxylate

[0331] Step 1: Synthesis of compound BnH12053

[0332] A solution of compound 3-1 (240 mg, 0.598 mmol), 2-bromo-1-methylimidazole (106 mg, 0.658 mmol), and Na2CO3 (127 mg, 1.20 mmol) in MeCN / HO (2:1, 7.72 mL) was vigorously stirred, bubbled with N2 gas for 5 min, and Pd(dppf)Cl2 (43.8 mg, 0.060 mmol) was added. The mixture was bubbled with N2 gas for 5 min and refluxed at 90 °C for 3 h. Subsequently, the mixture was cooled to room temperature, 1N NaOH (1.20 mL, 1.20 mmol) was added to the mixture, and stirred at room temperature for 3 h. The mixture was diluted with HO (3.00 mL) and washed with DCM (3.00 mL x 3). The aqueous layer was concentrated under reduced pressure and purified by column chromatography on C18-SiO2 (MeOH:H2O=1:1) to give compound BnH12053 (163 mg, 75%) as a white solid.

[0333] 1H NMR (400 MHz, DMSO-d6) δ 7.65 - 7.57 (m, 2H), 7.33 (d, J = 7.9 Hz, 1H), 7.28 (d, J = 7.9 Hz, 1H), 7.23 (s, 1H), 6.95 (s, 1H), 4.72 (s, 0.5H), 4.46 (dd, J = 14.6, 3.8 Hz, 0.5H), 4.20 (d, J = 5.7 Hz, 0.5H), 3.86 - 3.66 (m, 5.5H), 2.96 (t, J = 12.9 Hz, 0.5H), 2.55 (t, J = 12.9 Hz, 0.5H), 1.88 - 1.66 (m, 2H), 1.51 (tq, J = 31.1, 16.4, 13.9 Hz, 3H), 1.05 (t, J = 7.2 Hz, 3H).

[0334] (12) Synthesis of compound BnH12054

[0335] (3R,6S)-6-Methyl-1-(2-(4-(pyrimidin-4-yl)phenyl)acetyl)piperidine-3-carboxylic acid

[0336] Step 1: Synthesis of compound BnH12054

[0337] A solution of compound 3-1 (240 mg, 0.598 mmol), 4-chloropyrimidine (99.3 mg, 0.658 mmol), and Na2CO3 (190 mg, 1.79 mmol) in MeCN / H2O (2:1, 7.72 mL) was vigorously stirred, bubbled with N2 gas for 5 min, and Pd(dppf)Cl2 (43.8 mg, 0.060 mmol) was added. The mixture was bubbled with N2 gas for 5 min and refluxed at 90 °C for 3 h. Subsequently, the mixture was cooled to room temperature, 1N NaOH (1.20 mL, 1.20 mmol) was added to the mixture and stirred at room temperature for 3 h. The mixture was diluted with H2O (3.00 mL) and washed with DCM (3.00 mL × 3). The pH value was adjusted to pH 1-2 by adding 1N HCl to the aqueous layer, and the mixture was extracted with EtOAc (3.00 mL × 3). The combined organic layers were dried over MgSO4, filtered and concentrated under reduced pressure. The residue was purified by column chromatography on SiO2 (5% to 10% MeOH in DCM) to give compound BnH12054 (83.4 mg, 41%) as a white solid.

[0338] 1H NMR (400 MHz, DMSO-d6) δ 12.44 (s, 1H), 9.23 (s, 1H), 8.85 (d, J = 5.4 Hz, 1H), 8.17 (d, J = 8.0 Hz, 2H), 8.08 (d, J = 5.4 Hz, 1H), 7.40 (dd, J = 18.3, 7.9 Hz, 2H), 4.75 (s, 0.5H), 4.54 (dd, J = 14.0, 4.2 Hz, 0.5H), 4.28 (d, J = 8.3 Hz, 0.5H), 3.96 - 3.73 (m, 2.5H), 3.03 (t, J = 12.9 Hz, 0.5H), 2.62 (t, J = 12.7 Hz, 0.5H), 2.19 (tt, J = 12.0, 4.1 Hz, 1H), 1.79 (dt, J = 13.4, 4.5 Hz, 1H), 1.67 (tt, J = 17.4, 8.0 Hz, 1H), 1.51 (tq, J = 9.3, 4.6, 4.0 Hz, 2H), 1.08 (q, J = 8.3, 7.8 Hz, 3H).

[0339] (13) Synthesis of compound BnH12055

[0340] (3R,6S)-6-Methyl-1-(2-(4-(thiophen-2-yl)phenyl)acetyl)piperidine-3-carboxylic acid

[0341] Step 1: Synthesis of compound BnH12055

[0342] A solution of compound 3-1 (240 mg, 0.598 mmol), 2-bromothiophene (107 mg, 0.658 mmol), and Na2CO3 (190 mg, 1.79 mmol) in MeCN / H2O (2:1, 7.72 mL) was vigorously stirred, bubbled with N2 gas for 5 min, and Pd(dppf)Cl2 (43.8 mg, 0.060 mmol) was added. The mixture was bubbled with N2 gas for 5 min and refluxed at 90 °C for 3 h. Subsequently, the mixture was cooled to room temperature, 1N NaOH (1.20 mL, 1.20 mmol) was added to the mixture, and stirred at room temperature for 3 h. The mixture was diluted with H2O (3.00 mL) and washed with DCM (3.00 mL × 3). The pH value was adjusted to pH 1-2 by adding 1N HCl to the aqueous layer, and the mixture was extracted with EtOAc (3.00 mL × 3). The combined organic layers were dried over MgSO4, filtered and concentrated under reduced pressure. The residue was purified by column chromatography on SiO2 (5% MeOH in DCM) to give compound BnH12055 (30 mg, 14%) as an ivory solid.

[0343] 1H NMR (400 MHz, Chloroform-d) δ 7.57 (d, J = 7.9 Hz, 2H), 7.26 (d, J = 3.4 Hz, 4H), 7.07 (dd, J = 5.1, 3.6 Hz, 1H), 4.98 (s, 0.5H), 4.83 (d, J = 13.6 Hz, 0.5H), 4.18 (s, 0.5H), 3.93 (d, J = 14.1 Hz, 0.5H), 3.75 (s, 2H), 3.15 (t, J = 13.1 Hz, 0.5H), 2.78 (t, J = 12.9 Hz, 0.5H), 2.40 (s, 0.5H), 2.15 (s, 0.5H), 1.92 (d, J = 12.7 Hz, 1H), 1.79 (d, J = 12.3 Hz, 1H), 1.57 (d, J = 27.2 Hz, 2H), 1.14 (d, J = 6.9 Hz, 3H).

[0344] (14) Synthesis of compound BnH12056

[0345] (3R,6S)-1-(2-(4-(フラン-2-イル)フェニル)アセチル)-6-メチルピぺリジン-3-カルボン acid

[0346] Stage 1: Synthesis of compound BnH12056

[0347] A solution of compound 3-1 (240 mg, 0.598 mmol), 5-bromofuran (96.7 mg, 0.658 mmol), and Na2CO3 (190 mg, 1.79 mmol) in MeCN / H2O (2:1, 7.72 mL) was vigorously stirred, bubbled with N2 gas for 5 min, and Pd(dppf)Cl2 (43.8 mg, 0.060 mmol) was added. The mixture was bubbled with N2 gas for 5 min and refluxed at 90 °C for 3 h. Subsequently, the mixture was cooled to room temperature, 1N NaOH (1.20 mL, 1.20 mmol) was added to the mixture and stirred at room temperature for 3 h. The mixture was diluted with H2O (3.00 mL) and washed with DCM (3.00 mL × 3). The pH value was adjusted to pH 1-2 by adding 1N HCl to the aqueous layer, and the mixture was extracted with EtOAc (3.00 mL × 3). The combined organic layers were dried over MgSO4, filtered and concentrated under reduced pressure. The residue was purified by column chromatography on SiO2 (5% MeOH in DCM) to give compound BnH12056 (109 mg, 55%) as an ivory solid.

[0348] 1H NMR (400 MHz, Chloroform-d) δ 7.63 (d, J = 8.0 Hz, 2H), 7.48 - 7.43 (m, 1H), 7.27 (d, J = 6.0 Hz, 2H), 6.63 (d, J = 3.3 Hz, 1H), 6.46 (dd, J = 3.4, 1.8 Hz, 1H), 4.97 (s, 0.5H), 4.82 (d, J = 13.6 Hz, 0.5H), 4.17 (s, 0.5H), 3.92 (d, J = 13.7 Hz, 0.5H), 3.76 (s, 2H), 3.13 (t, J = 13.0Hz, 0.5H), 2.78 (t, J = 12.8 Hz, 0.5H), 2.39 (s, 0.5H), 2.11 (s, 0.5H), 1.91 (d, J = 12.7 Hz, 1H), 1.77 (d, J = 13.0 Hz, 1H), 1.67 - 1.40 (m, 2H), 1.13 (d, J = 8.7 Hz, 3H).

[0349] (15) Synthesis of compound BnH12072

[0350] (3R,6S)-6-Methyl-1-(2-(3'-nitro-[1,1'-biphenyl]-4-yl)acetyl)piperidine-3-carboxylic acid

[0351] Step 1: Synthesis of compound BnH12072

[0352] A solution of compound 3-1 (150 mg, 0.374 mmol), 3-bromonitrobenzene (75.5 mg, 0.374 mmol), and Na2CO3 (79.2 mg, 0.4748 mmol) in MeCN / HO (2:1, 4.82 mL) was vigorously stirred, bubbled with N2 gas for 5 min, and Pd(dppf)Cl2 (27.4 mg, 0.037 mmol) was added. The mixture was bubbled with N2 gas for 5 min and refluxed at 90 °C for 3 h. Subsequently, the mixture was cooled to room temperature, 1N NaOH (0.748 mL, 0.748 mmol) was added to the mixture, and stirred at room temperature for 17 h. The mixture was diluted with HO (4.00 mL) and washed with EtOAc (4.00 mL x 3). The aqueous layer was added with 1N HCl to adjust the pH value to pH 3-4, and the mixture was extracted with EtOAc (4.00 mL×3). The combined organic layers were dried over MgSO4, filtered, and concentrated under reduced pressure. The residue was purified by column chromatography on SiO2 (3%-5% MeOH in DCM) to give compound BnH12072 (120 mg, 84%) as an off-white solid.

[0353] 1H NMR (400 MHz, DMSO-d6) δ 8.44 (t, J = 2.0 Hz, 1H), 8.25 - 8.13 (m, 2H), 7.77 (t, J = 8.1 Hz, 3H), 7.38 (dd, J = 18.2, 7.9 Hz, 2H), 4.75 (s, 0.5H), 4.55 (d, J = 13.3 Hz, 0.5H), 4.29 (s, 0.5H), 3.94 (s, 0.5H), 3.85 - 3.73 (m, 2H), 3.05 (t, J = 12.8 Hz, 0.5H), 2.67 - 2.57 (t, J = 12.8 Hz, 0.5H), 2.20 (s, 1H), 1.80 (dd, J = 18.3, 6.8 Hz, 1H), 1.67 (d, J = 13.3 Hz, 1H), 1.54 (s, 2H), 1.08 (s, 3H).

[0354] (16) Synthesis of compound BnH12077

[0355] (3R,6S)-6-Methyl-1-(2-(4'-nitro-[1,1'-biphenyl]-4-yl)acetyl)piperidine-3-carboxylic acid

[0356] Step 1: Synthesis of compound BnH12077

[0357] A solution of compound 3-1 (150 mg, 0.374 mmol), 4-bromonitrobenzene (75.5 mg, 0.374 mmol), and Na2CO3 (79.2 mg, 0.748 mmol) in MeCN / HO (2:1, 4.82 mL) was vigorously stirred, bubbled with N2 gas for 5 min, and Pd(dppf)Cl2 (27.4 mg, 0.037 mmol) was added. The mixture was bubbled with N2 gas for 5 min and refluxed at 90 °C for 3 h. Subsequently, the mixture was cooled to room temperature, 1N NaOH (0.748 mL, 0.748 mmol) was added to the mixture, and stirred at room temperature for 17 h. The mixture was diluted with HO (4.00 mL) and washed with EtOAc (4.00 mL × 3). The aqueous layer was added with 1N HCl to adjust the pH value to pH 3-4, and the mixture was extracted with EtOAc (4.00 mL×3). The combined organic layers were dried over MgSO4, filtered, and concentrated under reduced pressure. The residue was purified by column chromatography on SiO2 (5%-10% MeOH in DCM) to give compound BnH12077 (107 mg, 75%) as a brown solid.

[0358] 1H NMR (400 MHz, DMSO-d6) δ 8.30 (d, J = 8.3 Hz, 2H), 7.97 (d, J = 8.4 Hz, 2H), 7.75 (d, J = 7.8 Hz, 2H), 7.38 (dd, J = 18.5, 7.8 Hz, 2H), 4.75 (s, 0.5H), 4.54 (dd, J = 13.1, 4.0 Hz, 0.5H), 4.29 (s, 0.5H), 3.92 (d, J = 12.9 Hz, 0.5H), 3.82 (q, J = 11.9, 8.6 Hz, 2H), 3.04 (t, J = 12.9 Hz, 0.5H), 2.62 (t, J = 12.8 Hz, 0.5H), 2.19 (d, J = 10.1 Hz, 1H), 1.79 (q, J = 9.5, 6.7 Hz, 1H), 1.66 (d, J = 13.2 Hz, 1H), 1.57 - 1.49 (m, 2H), 1.15 - 1.08 (m, 3H).

[0359] (17) Synthesis of compound BnH12081

[0360] (3R,6S)-1-(2-(4-(6-methoxypyridin-3-yl)phenyl)acetyl)-6-methylpiperidine-3-carboxylic acid

[0361] Step 1: Synthesis of compound BnH12081

[0362] A solution of compound 3-1 (220 mg, 0.548 mmol), 5-bromo-2-methoxypyridine (0.071 mL, 0.548 mmol), and Na2CO3 (116 mg, 1.10 mmol) in MeCN / HO (2:1, 7.07 mL) was vigorously stirred, bubbled with N2 gas for 5 min, and Pd(dppf)Cl2 (40.1 mg, 0.055 mmol) was added. The mixture was bubbled with N2 gas for 5 min and refluxed at 90 °C for 3 h. Subsequently, the mixture was cooled to room temperature, 1N NaOH (1.10 mL, 1.10 mmol) was added to the mixture, and stirred at room temperature for 17 h. The mixture was diluted with HO (7.00 mL) and washed with EtOAc (5.00 mL × 3). The aqueous layer was added with 1N HCl to adjust the pH value to pH 3-4, and the mixture was extracted with EtOAc (5.00 mL×3). The combined organic layers were dried over MgSO4, filtered, and concentrated under reduced pressure. The residue was purified by column chromatography on SiO2 (10% MeOH in DCM) to give compound BnH12081 (107 mg, 75%) as a white solid.

[0363] 1H NMR (400 MHz, DMSO-d6) δ 12.37 (s, 1H), 8.47 (d, J = 2.5 Hz, 1H), 8.00 (dd, J = 8.6, 2.6 Hz, 1H), 7.60 (d, J = 7.9 Hz, 2H), 7.30 (dd, J = 18.4, 7.8 Hz, 2H), 6.90 (d, J = 8.6 Hz, 1H), 4.75 (s, 0.5H), 4.54 (d, J = 13.5 Hz, 0.5H), 4.28 (s, 0.5H), 3.89 (s, 0.5H), 3.84 - 3.68 (m, 2H), 3.03 (t, J = 12.9 Hz, 0.5H), 2.67 - 2.56 (m, 0.5H), 2.19 (s, 1H), 1.85 - 1.74 (m, 1H), 1.66 (d, J = 13.1 Hz, 1H), 1.52 (s, 2H), 1.10 - 1.04 (m, 3H).

[0364] (18) Synthesis of compound BnH12082

[0365] (3R,6S)-1-(2-(4-(6-cyclopropylpyridin-3-yl)phenyl)acetyl)-6-methylpiperidine-3-carboxylic acid

[0366] Step 1: Synthesis of compound BnH12082

[0367] A solution of compound 3-1 (200 mg, 0.498 mmol), 3-bromo-6-(cyclopropyl)pyridine (0.060 mL, 0.498 mmol), and Na2CO3 (106 mg, 0.997 mmol) in MeCN / HO (2:1, 6.42 mL) was vigorously stirred, bubbled with N2 gas for 5 min, and Pd(dppf)Cl2 (36.5 mg, 0.050 mmol) was added. The mixture was bubbled with N2 gas for 5 min and refluxed at 90 °C for 2 h. Subsequently, the mixture was cooled to room temperature, 1N NaOH (1.10 mL, 1.10 mmol) was added to the mixture, and stirred at room temperature for 17 h. The mixture was diluted with HO (6.00 mL) and washed with EtOAc (4.00 mL × 3). The aqueous layer was added with 1N HCl to adjust the pH value to pH 1-2, and the mixture was washed with EtOAc (4.00 mL x 2). The aqueous layer was added with 1N NaOH to adjust the pH value to pH 4-5, and the mixture was extracted with EtOAc (4.00 mL x 3). The combined organic layers were dried over MgSO4, filtered, and concentrated under reduced pressure to give compound BnH12082 (120 mg, 64%) as a white solid.

[0368] 1H NMR (400 MHz, DMSO-d6) δ 12.30 (s, 1H), 8.70 (d, J = 2.4 Hz, 1H), 7.92 (dd, J = 8.1, 2.4 Hz, 1H), 7.63 (d, J = 7.8 Hz, 2H), 7.40 - 7.26 (m, 3H), 4.75 (s, 0.5H), 4.59 - 4.50 (m, 1=0.5H), 4.29 (s, 0.5H), 3.93 (d, J = 11.1 Hz, 0.5H), 3.85 - 3.69 (m, 2H), 3.04 (t, J = 12.8Hz, 0.5H), 2.68 - 2.57 (t, J = 12.8 Hz, 0.5H), 2.31 - 2.04 (m, 2H), 1.80 (dd, J = 13.2, 3.6 Hz, 1H), 1.66 (d, J = 12.5 Hz, 1H), 1.58 - 1.43 (m, 2H), 1.14 - 1.03 (m, 3H), 0.96 (td, J = 7.3, 6.5, 2.5 Hz, 4H).

[0369] (19) Synthesis of compound BnH12083

[0370] (3R,6S)-6-Methyl-1-(2-(4-(6-(trifluoromethyl)pyridin-3-yl)phenyl)acetyl)piperidine-3-carboxylic acid

[0371] Step 1: Synthesis of compound BnH12083

[0372] A solution of compound 3-1 (240 mg, 0.598 mmol), 5-bromo-2-trifluoromethylpyridine (135 mg, 0.598 mmol), and Na2CO3 (127 mg, 1.20 mmol) in MeCN / HO (2:1, 7.71 mL) was vigorously stirred, bubbled with N2 gas for 5 min, and Pd(dppf)Cl2 (43.8 mg, 0.060 mmol) was added. The mixture was bubbled with N2 gas for 5 min and refluxed at 90 °C for 3 h. Subsequently, the mixture was cooled to room temperature, 1N NaOH (1.20 mL, 1.20 mmol) was added to the mixture, and stirred at room temperature for 17 h. The mixture was diluted with HO (7.00 mL) and washed with EtOAc (5.00 mL × 3). The aqueous layer was added with 1N HCl to adjust the pH value to pH 3-4, and the mixture was extracted with EtOAc (5.00 mL×3). The combined organic layers were dried over MgSO4, filtered, and concentrated under reduced pressure. The residue was purified by column chromatography on SiO2 (10% MeOH in DCM) to give compound BnH12083 (167 mg, 69%) as an off-white solid.

[0373] 1H NMR (400 MHz, DMSO-d6) δ 12.41 (s, 1H), 9.10 (d, J = 2.3 Hz, 1H), 8.37 (dd, J = 8.2, 2.3 Hz, 1H), 7.98 (d, J = 8.2 Hz, 1H), 7.79 (d, J = 8.0 Hz, 2H), 7.40 (dd, J = 18.3, 7.9 Hz, 2H), 4.76 (s, 0.5H), 4.59 - 4.51 (d, J = 13.3 Hz, 0.5H), 4.34 - 4.25 (m, 0.5H), 3.94 (d, J = 13.3Hz, 0.5H), 3.82 (qd, J = 15.4, 5.8 Hz, 2H), 3.06 (t, J = 12.8 Hz, 0.5H), 2.69 - 2.58 (m, 0.5H), 2.31 - 2.13 (m, 1H), 1.87 - 1.77 (m, 1H), 1.67 (d, J = 11.6 Hz, 1H), 1.54 (dt, J = 6.7, 3.8 Hz, 2H), 1.14 - 1.02 (m, 3H).

[0374] (20) Synthesis of compound BnH12084

[0375] (3R,6S)-6-Methyl-1-(2-(4-(6-methylpyridin-3-yl)phenyl)acetyl)piperidine-3-carboxylic acid

[0376] Step 1: Synthesis of compound BnH12084

[0377] A solution of compound 3-1 (200 mg, 0.498 mmol), 5-bromo-2-methylpyridine (85.7 mg, 0.498 mmol), and Na2CO3 (106 mg, 0.997 mmol) in MeCN / HO (2:1, 6.42 mL) was vigorously stirred, bubbled with N2 gas for 5 min, and Pd(dppf)Cl2 (36.5 mg, 0.050 mmol) was added. The mixture was bubbled with N2 gas for 5 min and refluxed at 90 °C for 2.5 h. Subsequently, the mixture was cooled to room temperature, 1N NaOH (0.997 mL, 0.997 mmol) was added to the mixture, and stirred at room temperature for 3 h. The mixture was diluted with HO (6.00 mL) and washed with EtOAc (4.00 mL × 3). The aqueous layer was added with 1N HCl to adjust the pH value to pH 3-4, and the mixture was extracted with EtOAc (4.00 mL × 3). The combined organic layers were dried over MgSO4, filtered, and concentrated under reduced pressure. Et2O (2.00 mL) was added to the residue and stirred at room temperature for 30 min. The precipitate was filtered and washed with Et2O (3.00 mL) to give compound BnH12084 (92.0 mg, 52%) as a white solid.

[0378] 1H NMR 1H NMR (400 MHz, Chloroform-d) δ 8.82 (s, 1H), 8.02 (d, J = 7.9 Hz, 1H), 7.56 (d, J = 7.6 Hz, 2H), 7.41 (d, J = 7.5 Hz, 3H), 5.00 (s, 0.5H), 4.86 (s, 0.5H), 4.23 (s, 0.5H), 3.95 (d, J = 13.6 Hz, 0.5H), 3.84 (s, 2H), 3.18 (t, J = 12.8 Hz, 0.5H), 2.84 (s, 0.5H), 2.74 (s, 3H), 2.44 (s, 0.5H), 2.19 (s, 0.5H), 1.99 (d, J = 13.4 Hz, 1H), 1.82 (s, 1H), 1.64 (s, 2H), 1.17 (d, J = 7.9 Hz, 3H).

[0379] (21) Synthesis of compound BnH12085

[0380] (3R,6S)-1-(2-(4'-cyclohexyl-[1,1'-biphenyl]-4-yl)acetyl)-6-methylpiperidine-3-carboxylic acid

[0381] Step 1: Synthesis of compound BnH12085

[0382] A solution of compound 3-1 (191 mg, 0.476 mmol), 1-bromo-4-cyclohexylbenzene (0.088 mL, 0.476 mmol), and Na2CO3 (101 mg, 0.952 mmol) in MeCN / HO (2:1, 6.14 mL) was vigorously stirred, bubbled with N2 gas for 5 min, and Pd(dppf)Cl2 (34.8 mg, 0.048 mmol) was added. The mixture was bubbled with N2 gas for 5 min and refluxed at 90 °C for 2.5 h. Subsequently, the mixture was cooled to room temperature, 1N NaOH (1.20 mL, 1.20 mmol) was added to the mixture, and stirred at room temperature for 2 h. The mixture was diluted with HO (6.00 mL) and washed with EtOAc (4.00 mL × 3). The aqueous layer was added with 1N HCl to adjust the pH value to pH 2-3, and the mixture was extracted with EtOAc (4.00 mL × 3). The combined organic layers were dried over MgSO4, filtered, and concentrated under reduced pressure. The residue was purified by column chromatography on SiO2 (10% MeOH in DCM). Et2O (3.00 mL) was added to the residue and stirred at room temperature for 30 min. The precipitate was filtered and washed with Et2O (4.00 mL) to give compound BnH12085 (127 mg, 64%) as an off-white solid.

[0383] 1H NMR (400 MHz, DMSO-d6) δ 12.44 (s, 1H), 7.57 (t, J = 7.0 Hz, 4H), 7.28 (dd, J = 16.4, 7.9 Hz, 4H), 4.75 (s, 0.5H), 4.58 - 4.50 (m, 0.5H), 4.28 (s, 0.5H), 3.92 (d, J = 13.6 Hz, 0.5H), 3.86 - 3.67 (m, 2H), 3.04 (t, J = 12.8 Hz, 0.5H), 2.63 ((t, J = 12.8 Hz, 0.5H), 2.27 - 2.12 (m, 1H), 1.91 - 1.15 (m, 15H), 1.07 (t, J = 6.4 Hz, 3H).

[0384] (22) Synthesis of compound BnH12086

[0385] (3R,6S)-1-(2-(4'-amino-[1,1'-biphenyl]-4-yl)acetyl)-6-methylpiperidine-3-carboxylic acid

[0386] Step 3: Synthesis of compound BnH12086

[0387] A solution of compound 3-1 (225 mg, 0.561 mmol), 4-bromoaniline (96.5 mg, 0.561 mmol), and Na2CO3 (119 mg, 1.12 mmol) in MeCN / H2O (2:1, 7.01 mL) was vigorously stirred, bubbled with N2 gas for 5 min, and Pd(dppf)Cl2 (41.0 mg, 0.06 mmol) was added. The mixture was bubbled with N2 gas for 5 min and refluxed at 90 °C for 2 h. Subsequently, the mixture was cooled to room temperature, 1N NaOH (1.12 mL, 1.12 mmol) was added to the mixture, and stirred at room temperature for 4 h. The mixture was diluted with H2O (5.00 mL) and washed with EtOAc (5.00 mL × 3). The pH value was adjusted to pH 1-2 by adding 1N HCl to the aqueous layer, and the mixture was washed with EtOAc (5.00 mL × 2). The aqueous layer was added with 1N NaOH to adjust the pH value to pH 5-6, and the mixture was extracted with EtOAc (5.00 mL × 3). The combined organic layers were dried over MgSO4, filtered, and concentrated under reduced pressure. The residue was purified by column chromatography on SiO2 (0-10% MeOH in DCM). Et2O (2.00 mL) was added to the residue and stirred at room temperature for 30 min. The precipitate was filtered and washed with Et2O (2.00 mL) to give compound BnH12086 (70.0 mg, 35%) as a beige solid.

[0388] 1H NMR (400 MHz, DMSO-d6) δ 12.44 (s, 1H), 7.49-7.46 (m, 2H), 7.36-7.34 (d, J = 8.2 Hz, 2H), 7.24-7.17 (dd, J = 18.4, 7.8 Hz, 2H), 6.64-6.62 (d, J = 8.2 Hz, 2H), 5.20 (s, 2H), 4.75 (s, 0.5H), 4.56-4.52 (m, 0.5H), 4.26 (s, 0.5H), 3.92-3.89 (d, J = 13.7 Hz, 0.5H), 3.77-3.66 (m, 2H), 3.04-2.98 (t, J = 12.8 Hz, 0.5H), 2.64-2.57 (t, J = 12.9 Hz, 0.5H), 2.16-2.12 (d, J = 13.6 Hz, 1H), 1.80-1.77 (d, J = 12.9 Hz, 1H), 1.66-1.63 (d, J = 13.0 Hz, 1H), 1.53-1.50 (d, J = 10.9 Hz, 2H), 1.07-1.05 (dd, J = 6.9, 2.3 Hz, 3H).

[0389] (23) Synthesis of compound BnH12087

[0390] (3R,6S)-6-Methyl-1-(2-(4-(6-phenylpyridin-3-yl)phenyl)acetyl)piperidine-3-carboxylic acid

[0391] Step 3: Synthesis of compound BnH12087

[0392] A solution of compound 3-1 (194 mg, 0.483 mmol), 5-bromo-2-phenylpyridine (113 mg, 0.483 mmol), and Na2CO3 (102 mg, 0.967 mmol) in MeCN / HO (2:1, 6.04 mL) was vigorously stirred, bubbled with N2 gas for 5 min, and Pd(dppf)Cl2 (35.4 mg, 0.05 mmol) was added. The mixture was bubbled with N2 gas for 5 min and refluxed at 90 °C for 2 h. Subsequently, the mixture was cooled to room temperature, 1N NaOH (0.967 mL, 0.967 mmol) was added to the mixture, and stirred at room temperature for 4 h. The mixture was diluted with HO (5.00 mL) and washed with EtOAc (5.00 mL x 3). The aqueous layer was added with 1N HCl to adjust the pH value to pH 1-2, and the mixture was extracted with EtOAc (4.00 mL × 3). The combined organic layers were dried over MgSO4, filtered, and concentrated under reduced pressure. The residue was purified by column chromatography on SiO2 (0-4% MeOH in DCM). Et2O (2.00 mL) was added to the residue and stirred at room temperature for 30 min. The precipitate was filtered and washed with Et2O (2.00 mL) to give compound BnH12087 (70.1 mg, 35%) as an off-white solid.

[0393] 1H NMR (400 MHz, DMSO-d6) δ 8.99 (s, 1H), 8.18-8.13 (t, J = 8.7 Hz, 3H), 8.06-8.04 (d, J = 8.4 Hz, 1H), 7.76-7.73 (dd, J = 8.0, 4.4 Hz, 2H), 7.53-7.49 (t, J = 7.5 Hz, 2H), 7.47-7.43 (t, J = 7.2 Hz, 1H), 7.39-7.37 (d, J = 7.8 Hz, 1H), 7.34-7.32 (d, J = 7.9 Hz, 1H), 4.74 (s, 0.5H), 4.55-4.50 (m, 0.5H), 4.28 (s, 0.5H), 3.92-3.89 (d, J = 13.8 Hz, 0.5H), 3.83-3.76 (m, 2H), 3.04-2.98 (t, J = 12.9 Hz, 0.5H), 2.63-2.56 (d, J = 12.6 Hz, 0.5H), 2.11-2.08 (d, J = 10.2 Hz, 1H), 1.80-1.76 (d, J = 12.8 Hz, 1H), 1.64-1.61 (m, 1H), 1.51 (s, 2H), 1.10-1.05 (dd, J = 14.4, 6.8 Hz, 3H).

[0394] (24) Synthesis of compound BnH12088

[0395] (3R,6S)-1-(2-(3'-アミノ-[1,1'-ビフェニル]-4-イル)アセチル)-6-メチルピペリジン-3-カルボン acid

[0396] Stage 1: Synthesis of compound BnH12088

[0397] A solution of compound 3-1 (210 mg, 0.523 mmol), 3-bromoaniline (0.057 mL, 0.523 mmol), and Na2CO3 (111 mg, 1.05 mmol) in MeCN / HO (2:1, 6.75 mL) was vigorously stirred, bubbled with N2 gas for 5 min, and Pd(dppf)Cl2 (38.3 mg, 0.052 mmol) was added. The mixture was bubbled with N2 gas for 5 min and refluxed at 90 °C for 2.5 h. Subsequently, the mixture was cooled to room temperature, 1N NaOH (1.05 mL, 1.05 mmol) was added to the mixture, and stirred at room temperature for 3 h. The mixture was diluted with HO (6.00 mL) and washed with EtOAc (4.00 mL × 3). The aqueous layer was added with 1N HCl to adjust the pH value to pH 1-2, and the mixture was washed with EtOAc (4.00 mL x 2). The aqueous layer was added with 1N NaOH to adjust the pH value to pH 5-6, and the mixture was extracted with EtOAc (4.00 mL x 3). The combined organic layers were dried over MgSO4, filtered, and concentrated under reduced pressure. Et2O / n-Hexane (2:1, 2.00 mL) was added to the residue and stirred at room temperature for 30 min. The precipitate was filtered and washed with Et2O / n-Hexane (2:1, 3.00 mL) to give compound BnH12088 (115 mg, 62%) as a beige solid.

[0398] 1H NMR (400 MHz, DMSO-d6) δ 12.43 (s, 1H), 7.49 (d, J = 7.7 Hz, 2H), 7.26 (dd, J = 18.0, 7.8 Hz, 2H), 7.08 (t, J = 7.8 Hz, 1H), 6.83 (d, J = 2.1 Hz, 1H), 6.76 (d, J = 7.7 Hz, 1H), 6.54 (dd, J = 7.9, 2.2 Hz, 1H), 5.13 (s, 2H), 4.75 (s, 0.5H), 4.55 (d, J = 13.6 Hz, 0.5H), 4.27 (s, 0.5H), 3.90 (s, 0.5H), 3.84 - 3.61 (m, 2H), 3.03 (t, J = 12.8 Hz, 0.5H), 2.66 (t, J = 12.8 Hz, 0.5H), 2.18 (s, 1H), 1.79 (d, J = 12.7 Hz, 1H), 1.66 (d, J = 12.6 Hz, 1H), 1.52 (d, J = 8.8 Hz, 2H), 1.08 (q, J = 6.5, 6.1 Hz, 3H).

[0399] (25) Synthesis of compound BnH12089

[0400] (3R,6S)-6-メチル-1-(2-(4'-(ピリジン-3-イル)- [1,1'-ビフェニル]-4-イル)アセチル)ピペリジン-3-カルボン acid

[0401] Stage 1: Synthesis of compound BnH12089

[0402] A solution of compound 3-1 (201 mg, 0.501 mmol), 3-(4-bromophenyl)pyridine (117 mg, 0.501 mmol), and Na2CO3 (106 mg, 1.00 mmol) in MeCN / HO (2:1, 6.46 mL) was vigorously stirred, bubbled with N2 gas for 5 min, and Pd(dppf)Cl2 (36.7 mg, 0.050 mmol) was added. The mixture was bubbled with N2 gas for 5 min and refluxed at 90 °C for 2 h. Subsequently, the mixture was cooled to room temperature, 1N NaOH (1.00 mL, 1.00 mmol) was added to the mixture, and stirred at room temperature for 3 h. The mixture was diluted with HO (6.00 mL) and washed with EtOAc (4.00 mL × 3). The aqueous layer was added with 1N HCl to adjust the pH value to pH 1-2, and the mixture was washed with EtOAc (4.00 mL x 2). The aqueous layer was added with 1N NaOH to adjust the pH value to pH 4-5, and the mixture was extracted with EtOAc (4.00 mL x 3). The combined organic layers were dried over MgSO4, filtered, and concentrated under reduced pressure. Et2O (3.00 mL) was added to the residue and stirred at room temperature for 30 min. The precipitate was filtered and washed with Et2O (4.00 mL) to give compound BnH12089 (127 mg, 61%) as a white solid.

[0403] 1H NMR (400 MHz, DMSO-d6) δ 12.43 (s, 1H), 8.96 (d, J = 2.4 Hz, 1H), 8.59 (dd, J = 4.8, 1.6 Hz, 1H), 8.14 (dt, J = 8.0, 2.0 Hz, 1H), 7.84 (t, J = 6.4 Hz, 4H), 7.70 (d, J = 7.8 Hz, 2H), 7.51 (dd, J = 7.9, 4.7 Hz, 1H), 7.34 (dd, J = 18.2, 7.8 Hz, 2H), 4.76 (s, 0.5H), 4.56 (d, J = 13.8 Hz, 0.5H), 4.30 (s, 0.5H), 4.00 - 3.90 (d, J = 13.8 Hz, 0.5H), 3.87 - 3.71 (m, 2H), 3.05 (t, J = 12.9 Hz, 0.5H), 2.68 - 2.58 (t, J = 12.9 Hz, 0.5H), 2.20 (d, J = 3.9 Hz, 1H), 1.81 (d, J = 12.7 Hz, 1H), 1.75 - 1.60 (m, 1H), 1.54 (s, 2H), 1.09 (dd, J = 10.5, 6.8 Hz, 3H).

[0404] (26) Synthesis of compound BnH12090

[0405] (3R,6S)-1-(2-(4'-(ジエチルアミノ)-[1,1'-ビフェニル]-4-イル)アセチル)-6-メチルピペリジン-3-カルボン acid

[0406] Stage 1: Synthesis of compound BnH12090

[0407] A solution of compound 3-1 (252 mg, 0.628 mmol), 4-bromo-N,N-diethylaniline (143 mg, 0.628 mmol), and Na2CO3 (106 mg, 1.00 mmol) in MeCN / HO (2:1, 7.85 mL) was vigorously stirred, bubbled with N2 gas for 5 min, and Pd(dppf)Cl2 (46.0 mg, 0.06 mmol) was added. The mixture was bubbled with N2 gas for 5 min and refluxed at 90 °C for 2 h. Subsequently, the mixture was cooled to room temperature, 1N NaOH (1.00 mL, 1.00 mmol) was added to the mixture, and stirred at room temperature for 4 h. The mixture was diluted with HO (6.00 mL) and washed with EtOAc (4.00 mL × 3). The aqueous layer was added with 1N HCl to adjust the pH value to pH 4-5, and the mixture was extracted with EtOAc (4.00 mL × 3). The combined organic layers were dried over MgSO4, filtered, and concentrated under reduced pressure. The residue was purified by column chromatography on SiO2 (5-10% MeOH in DCM). Et2O (2.00 mL) was added to the residue and stirred at room temperature for 30 min. The precipitate was filtered and washed with Et2O (3.00 mL) to give compound BnH12090 (55.0 mg, 21%) as an off-white solid.

[0408] 1H NMR (400 MHz, DMSO-d6) δ 12.47 (s, 1H), 7.52-7.47 (dd, J = 13.4, 7.9 Hz, 4H), 7.26-7.19 (dd, J = 18.3, 7.8 Hz, 2H), 6.74-6.72 (d, J = 8.6 Hz, 2H), 4.75 (s, 0.5H), 4.57-4.53 (d, J = 13.9 Hz, 0.5H), 4.28 (s, 0.5H), 3.94-3.90 (d, J = 14.1 Hz, 0.5H), 3.78-3.67 (m, 2H), 3.48-3.34 (m, 4H), 3.05-2.99 (t, J = 12.8 Hz, 0.5H), 2.64-2.58 (t, J = 12.8 Hz, 0.5H), 2.20-2.14 (m, 1H), 1.81-1.77 (m, 1H), 1.70-1.64 (d, J = 12.6 Hz, 1H), 1.53-1.51 (d, J = 9.3 Hz, 2H), 1.13-1.10 (t, J = 6.9 Hz, 6H), 1.08-1.05 (dd, J = 7.1, 2.9 Hz, 3H).

[0409] Preparation Example 3. Preparation of active compound according to representative reaction scheme 4

[0410] [Reaction Scheme 4]

[0411] JPEG2025512899000024.jpg26143

[0412] JPEG2025512899000025.jpg65131

[0413] (1) Synthesis of compound BnH12015

[0414] (R)-N-Hydroxy-1-(2-(4-(pyrimidin-5-yl)phenyl)acetyl)pyrrolidine-2-carboxamide

[0415] Step 1: Synthesis of 2-(4-(pyrimidin-5-yl)phenyl)acetic acid

[0416] A suspension of 4-bromophenylacetic acid (1.00 g, 4.65 mmol), 5-pyrimidineboronic acid pinacol ester (910 mg, 4.42 mmol), and Na2CO3 (986 mg, 9.30 mmol) in MeCN / H2O (2:1, 60.0 mL) was vigorously stirred and bubbled with N2 gas for 5 min, followed by the addition of Pd(dppf)Cl2 (340 mg, 0.465 mmol). The mixture was bubbled with N2 gas for 5 min and refluxed at 90 °C for 2 h. After completion of the reaction, the reaction mixture was filtered through a Celite pad and washed with EtOAc (10.0 mL × 2). The filtrate was diluted with H2O (40.0 mL) and washed with EtOAc (40.0 mL × 2). The aqueous layer was acidified with 1 N HCl to pH 1-2 and stirred for 30 min. The precipitate was filtered and washed with H2O (40.0 mL). The filter cake was dried under vacuum to give the desired compound (542 mg, 54%) as a white solid.

[0417] Step 2: Synthesis of methyl (2-(4-(pyrimidin-5-yl)phenyl)acetyl)-D-prolinate

[0418] To 2-(4-(pyrimidin-5-yl)phenyl)acetic acid (110 mg, 0.513 mmol) in DMF (1.71 mL) was added methyl D-prolinate (10.2 mg, 0.616 mmol), DIPEA (0.135 mL, 0.770 mmol), HOBt (83.3 mg, 0.616 mmol), and EDC·HCl (95.7 mg, 0.616 mmol). The mixture was stirred at room temperature for 3 h 3 min. The mixture was diluted with EtOAc (5.00 mL) and washed with saturated aqueous NH4Cl (5.00 mL) and brine (5.00 mL). The organic layer was dried over MgSO4, filtered, and concentrated under reduced pressure. The residue was purified by column chromatography on SiO2 (n-hexane:EtOAc = 2:1) to give the desired compound (145 mg, 79%) as a colorless oil.

[0419] Step 3: Synthesis of compound BnH12015

[0420] To a solution of methyl (2-(4-(pyrimidin-5-yl)phenyl)acetyl)-D-prolinate (83.0 mg, 0.255 mmol) in THF / MeOH (1:1, 3.00 mL) was added hydroxylamine hydrochloride (169 mg, 2.55 mmol) and KOH (28.6 mg, 0.510 mmol). The mixture was stirred at room temperature for 2 h. The mixture was diluted with H2O (2.00 mL) and washed twice with DCM (3.00 mL x 2). The aqueous layer was acidified with 1N HCl at 0 °C until pH 1-2 and extracted with EtOAc (3.00 mL). The organic layer was washed with brine (3.00 mL), dried over MgSO4, filtered, and concentrated under reduced pressure. The residue was purified by column chromatography on SiO2 (DCM:MeOH=9:1) to give compound BnH12015 (48.0 mg, 57%) as a white solid.

[0421] 1H NMR (400 MHz, Chloroform-d) δ 9.90 (s, 1H), 9.21 (s, 1H), 8.94 (s, 2H), 7.60 - 7.53 (m, 2H), 7.40 (d, J = 8.0 Hz, 2H), 4.55 (d, J = 8.0 Hz, 1H), 3.75 (s, 2H), 3.65 - 3.59 (m, 1H), 3.52 (q, J = 8.7 Hz, 1H), 2.49 (s, 1H), 2.25 - 2.12 (m, 1H), 2.00 (d, J = 38.2 Hz, 2H).

[0422] (2) Synthesis of compound BnH12017

[0423] Step 1: Synthesis of methyl (2-(4-(pyrimidin-5-yl)phenyl)acetyl)-D-prolinate

[0424] To 2-(4-(pyrimidin-5-yl)phenyl)acetic acid (110 mg, 0.513 mmol) in DMF (1.71 mL) was added methyl L-prolinate (10.2 mg, 0.616 mmol), DIPEA (0.135 mL, 0.770 mmol), HOBt (83.3 mg, 0.616 mmol), and EDC·HCl (95.7 mg, 0.616 mmol). The mixture was stirred at room temperature for 3 h 3 min. The mixture was diluted with EtOAc (5.00 mL) and washed with saturated aqueous NH4Cl (5.00 mL) and brine (5.00 mL). The organic layer was dried over MgSO4, filtered, and concentrated under reduced pressure. The residue was purified by column chromatography on SiO2 (n-hexane:EtOAc = 2:1) to give the desired compound (83 mg, 49%) as a colorless oil.

[0425] Step 2: Synthesis of compound BnH12017

[0426] To a solution of methyl (2-(4-(pyrimidin-5-yl)phenyl)acetyl)-D-prolinate (80.0 mg, 0.246 mmol) in THF / MeOH (1:1, 3.00 mL) was added hydroxylamine hydrochloride (162 mg, 2.46 mmol) and KOH (27.6 mg, 0.492 mmol). The mixture was stirred at room temperature for 2 h. The mixture was diluted with H2O (2.00 mL) and washed twice with DCM (3.00 mL x 2). The aqueous layer was acidified with 1N HCl at 0 °C until pH 1-2 and extracted with EtOAc (3.00 mL). The organic layer was washed with brine (3.00 mL), dried over MgSO4, filtered, and concentrated under reduced pressure. The residue was purified by column chromatography on SiO2 (DCM:MeOH=9:1) to give compound BnH12017 (52.0 mg, 64%) as a white solid.

[0427] 1H NMR (400 MHz, Chloroform-d) δ 9.87 (s, 1H), 7.35 - 7.27 (m, 2H), 7.15 (dd, J = 8.4, 2.6 Hz, 2H), 6.99 (td, J = 8.3, 2.7 Hz, 2H), 4.54 (d, J = 8.1 Hz, 1H), 3.73 (d, J = 2.6 Hz, 2H), 3.66 - 3.49 (m, 1H), 2.46 (s, 1H), 2.17 (dd, J = 17.6, 8.5 Hz, 1H), 2.09 - 1.89 (m, 2H).

[0428] Preparation Example 4. Preparation of active compound according to representative reaction scheme 5

[0429] [Reaction Scheme 5]

[0430] JPEG2025512899000026.jpg34147

[0431] JPEG2025512899000027.jpg36136

[0432] (1) Synthesis of compound BnH12058

[0433] (3R,6S)-6-Methyl-1-(2-(4-(pyrimidin-2-yl)phenyl)acetyl)piperidine-3-carboxamide

[0434] Step 1: Synthesis of compound BnH12058

[0435] To a suspension of compound BnH12048 (100 mg, 0.295 mmol) in MeCN (3.17 mL) was added CDI (71.7 mg, 0.442 mmol) and stirred at room temperature for 20 min. To the mixture was added NH4OH (0.172 mL, 4.42 mmol) and the mixture was stirred at room temperature for 17 h. The mixture was diluted with EtOAc (4.00 mL) and washed with saturated NH4Cl (4.00 mL), 10% citric acid (4.00 mL), and brine (4.00 mL). The organic layer was dried over MgSO4, filtered, and concentrated. The residue was purified by column chromatography (5% MeOH in DCM) to give compound BnH12058 (48 mg, 48%) as a white solid.

[0436] 1H NMR (400 MHz, DMSO-d6) δ 8.97 (d, J = 0.8 Hz, 2H), 8.31 - 8.24 (m, 2H), 7.43 - 7.29 (m, 3H), 6.86 (d, J = 9.7 Hz, 1H), 4.74 (d, J = 6.7 Hz, 0.5H), 4.38 (dd, J = 13.3, 4.2 Hz, 0.5H), 4.30 - 4.24 (m, 0.5H), 3.87 - 3.75 (m, 2.5H), 3.02 (dd, J = 13.7, 11.9 Hz, 0.5H), 2.61 (q, J = 14.0, 13.0 Hz, 0.5H), 2.08 (ddd, J = 16.4, 10.5, 3.1 Hz, 1H), 1.68 (td, J = 13.0, 11.4, 4.6 Hz, 2H), 1.56 - 1.39 (m, 2H), 1.12 - 1.02 (m, 3H).

[0437] (2) Synthesis of compound BnH12059

[0438] (3R,6S)-1-(2-(4-(5-fluoropyrimidin-2-yl)phenyl)acetyl)-6-methylpiperidine-3-carboxamide

[0439] Step 1: Synthesis of compound BnH12059

[0440] To a suspension of compound BnH12049 (100 mg, 0.280 mmol) in MeCN (3.01 mL) was added CDI (68.1 mg, 0.420 mmol) and stirred at room temperature for 20 min. To the mixture was added NH4OH (0.163 mL, 4.20 mmol) and the mixture was stirred at room temperature for 17 h. The mixture was diluted with EtOAc (4.00 mL) and washed with saturated NH4Cl (4.00 mL), 10% citric acid (4.00 mL), and brine (4.00 mL). The organic layer was dried over MgSO4, filtered, and concentrated. The residue was purified by column chromatography (5% MeOH in DCM) to give compound BnH12059 (52 mg, 52%) as a white solid.

[0441] 1H NMR (400 MHz, DMSO-d6) δ 8.89 (d, J = 4.9 Hz, 2H), 8.34 (dd, J = 8.4, 2.3 Hz, 2H), 7.47 - 7.29 (m, 4H), 6.86 (d, J = 9.5 Hz, 1H), 4.75 (s, 0.5H), 4.38 (dd, J = 13.0, 4.2 Hz, 0.5H), 4.25 (d, J = 6.6 Hz, 0.5H), 3.95 - 3.72 (m, 2.5H), 3.02 (t, J = 12.8 Hz, 0.5H), 2.63 (t, J = 12.7Hz, 0.5H), 2.09 (dd, J = 13.6, 9.5 Hz, 1H), 1.66 (d, J = 14.3 Hz, 2H), 1.58 - 1.36 (m, 2H), 1.13 - 1.02 (m, 3H).

[0442] Preparation Example 5. Preparation of active compounds according to representative reaction scheme 6

[0443] [Reaction Scheme 6]

[0444] JPEG2025512899000028.jpg59140

[0445] JPEG2025512899000029.jpg71131

[0446] (1) Synthesis of compound BnH12064

[0447] (S)-1-(2-Methylpiperidin-1-yl)-2-(4-(pyridin-3-yl)phenyl)ethan-1-one

[0448] Step 1: Synthesis of 2-(4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenyl)acetic acid

[0449] A suspension of compounds 2-(4-bromophenyl)acetic acid (700 mg, 3.26 mmol), bis(pinacolato)diborane (911 mg, 3.59 mmol), and KOAc (958.4 mg, 9.77 mmol) in 1,4-dioxane (11.2 mL) was vigorously stirred and bubbled with N2 gas for 5 min, followed by the addition of Pd(dppf)Cl2 (238 mg, 0.326 mmol). The mixture was bubbled with N2 gas for 5 min and refluxed at 100 °C for 17 h. After completion of the reaction, the reaction mixture was filtered through a celite pad and washed with DCM (5.00 mL × 3). The combined organic layer was washed with H2O (20.0 mL), dried over MgSO4, filtered, and concentrated under reduced pressure. The residue was purified by column chromatography on SiO2 (n-hexane:EtOAc = 3:1 to 2:1) to give compound 6-1-BnH12064-1 (552 mg, 65%) as a yellow solid.

[0450] Step 2: Synthesis of 2-(4-(pyridin-3-yl)phenyl)acetic acid (6-1-BnH12064)

[0451] A solution of compound 6-1-BnH12064-1 (450 mg, 1.72 mmol), 3-bromopyridine (0.182 mL, 1.89 mmol), and Na2CO3 (546 mg, 5.15 mmol) in MeCN / H2O (2:1, 21.4 mL) was vigorously stirred, bubbled with N2 gas for 5 min, and Pd(dppf)Cl2 (126 mg, 0.172 mmol) was added. The mixture was bubbled with N2 gas for 5 min and refluxed at 90 °C for 3 h. Then, the mixture was cooled to room temperature. The mixture was diluted with H2O (10.0 mL) and washed with DCM (10.0 mL × 3). The pH value was adjusted to pH 1-2 by adding 1 N HCl to the aqueous layer, and the mixture was extracted with EtOAc (15.0 mL × 3). The combined organic layers were dried over MgSO4, filtered, concentrated under reduced pressure and used without further purification.

[0452] Step 3: Synthesis of compound BnH12064

[0453] To a solution of 2-(4-(pyridin-3-yl)phenyl)acetic acid (6-1-BnH12064, 90 mg, 0.422 mmol) in DMF (1.41 mL) was added (S)-2-methylpiperidine (44.0 mg, 0.443 mmol), DIPEA (0.110 mL, 0.633 mmol), HOBt (68.4 mg, 0.506 mmol), and EDC·HCl (78.6 mg, 0.506 mmol). The mixture was stirred at room temperature for 3 h. The mixture was diluted with EtOAc (2.00 mL) and washed with saturated aqueous NH4Cl (2.00 mL) and brine (2.00 mL). The organic layer was dried over MgSO4, filtered, and concentrated under reduced pressure. The residue was purified by column chromatography on SiO2 (n-hexane:EtOAc = 1:1 to 0:1) to give compound BnH12064 (46.6 mg, 38%) as a yellowish oil.

[0454] 1H NMR (400 MHz, Chloroform-d) δ 8.88 (d, J = 2.3 Hz, 1H), 8.62 (dd, J = 5.0, 1.6 Hz, 1H), 8.00 (dt, J = 8.0, 2.0 Hz, 1H), 7.57 (d, J = 7.9 Hz, 2H), 7.47 (dd, J = 8.0, 4.9 Hz, 1H), 7.39 (d, J = 8.9 Hz, 2H), 4.99 (s, 0.5H), 4.58 (d, J = 13.7 Hz, 0.5H), 4.22 (s, 0.5H), 3.80 (s, 2H), 3.68 (d, J = 13.8 Hz, 0.5H), 3.11 (t, J = 13.5 Hz, 0.5H), 2.72 (t, J = 13.6 Hz, 0.5H), 1.58 (d, J = 31.7 Hz, 6H), 1.18 (d, J = 6.9 Hz, 3H).

[0455] Preparation Example 6. Preparation of active compound according to representative reaction scheme 7 (anhydrous form)

[0456] [Reaction Scheme 7]

[0457] JPEG2025512899000030.jpg38146

[0458] JPEG2025512899000031.jpg117131

[0459] (1) Synthesis of compound BnH12066

[0460] (3S,6R)-6-Methyl-1-(2-(4-(pyridin-3-yl)phenyl)acetyl)piperidine-3-carboxylic acid

[0461] Step 1: Synthesis of methyl (3S,6R)-6-methyl-1-(2-(4-(pyridin-3-yl)phenyl)acetyl)piperidine-3-carboxylate

[0462] To a solution of 2-(4-(pyridin-3-yl)phenyl)acetic acid (6-1-BnH12064, 90 mg, 0.422 mmol) in DMF (1.41 mL) was added methyl (3S,6R)-6-methylpiperidine-3-carboxylate (anhydrous form) (69.7 mg, 0.443 mmol), DIPEA (0.110 mL, 0.633 mmol), HOBt (68.4 mg, 0.506 mmol), and EDC·HCl (78.6 mg, 0.506 mmol). The mixture was stirred at room temperature for 3 h. The mixture was diluted with EtOAc (2.00 mL) and washed with saturated aqueous NH4Cl (2.00 mL) and brine (2.00 mL). The organic layer was dried over MgSO4, filtered, and concentrated under reduced pressure. The residue was purified by column chromatography on SiO.sub.2 (5% MeOH in DCM) to give the desired compound (37.0 mg, 25%) as a yellowish oil.

[0463] Step 2: Synthesis of compound BnH12066

[0464] To a solution of methyl (3S,6R)-6-methyl-1-(2-(4-(pyridin-3-yl)phenyl)acetyl)piperidine-3-carboxylate (37.0 mg, 0.108 mmol) in THF (0.373 mL) was added 1N NaOH (0.162 mL, 0.162 mmol). The mixture was stirred at room temperature for 2 h. The mixture was diluted with H2O (2.00 mL) and washed twice with DCM (2.00 mL x 2). The aqueous layer was acidified with 1N HCl at 0 °C until pH 1-2 and extracted with EtOAc (3.00 mL). The organic layer was washed with brine (3.00 mL), dried over MgSO4, filtered, and concentrated under reduced pressure to provide compound BnH12066 (30.0 mg, 82%) as a white solid.

[0465] 1H NMR (400 MHz, DMSO-d6) δ 9.08 (d, J = 2.2 Hz, 1H), 8.72 (d, J = 5.2 Hz, 1H), 8.50 (d, J = 7.9 Hz, 1H), 7.80 (dd, J = 19.9, 8.1 Hz, 3H), 7.40 (dd, J = 18.3, 7.9 Hz, 2H), 4.75 (s, 0.5H), 4.58 - 4.51 (m, 0.5H), 4.30 (s, 0.5H), 3.93 (d, J = 13.6 Hz, 0.5H), 3.89 - 3.74 (m, 2H), 3.05 (t, J = 12.8 Hz, 0.5H), 2.61 (t, J = 12.8 Hz, 0.5H), 2.22 (d, J = 13.8 Hz, 1H), 1.81 (d, J = 12.7 Hz, 1H), 1.67 (d, J = 12.6 Hz, 1H), 1.58 - 1.46 (m, 2H), 1.09 (dd, J = 16.1, 6.9 Hz, 3H).

[0466] (2) Synthesis of compound BnH12071

[0467] (3S,6R)-6-Methyl-1-(2-(4-(pyrimidin-5-yl)phenyl)acetyl)piperidine-3-carboxylic acid

[0468] Step 1: Synthesis of methyl (3S,6R)-6-methyl-1-(2-(pyridin-3-yl)acetyl)piperidine-3-carboxylate

[0469] To a solution of 2-(4-(pyrimidin-5-yl)phenyl)acetic acid (100 mg, 0.467 mmol) in DMF (2.00 mL) was added methyl (3S,6R)-6-methylpiperidine-3-carboxylate (anhydrous form) (77.1 mg, 0.490 mmol), DIPEA (0.122 mL, 0.700 mmol), HOBt (75.7 mg, 0.560 mmol), and EDC·HCl (107 mg, 0.560 mmol). The mixture was stirred at room temperature for 2 h. The mixture was diluted with EtOAc (3.00 mL) and washed with saturated aqueous NH4Cl (3.00 mL) and brine (3.00 mL). The organic layer was dried over MgSO4, filtered, and concentrated under reduced pressure. The residue was purified by column chromatography on SiO2 (EtOAc alone) to give the desired compound (93.5 mg, 57%) as a pale yellow oil.

[0470] Step 2: Synthesis of compound BnH12071

[0471] To a solution of methyl (3S,6R)-6-methyl-1-(2-(4-(pyrimidin-5-yl)phenyl)acetyl)piperidine-3-carboxylate (92.0 mg, 0.260 mmol) in THF (1.00 mg, 0.260 mmol) was added 1N NaOH (0.390 mL, 0.390 mmol). The mixture was stirred at room temperature for 2 h. The mixture was diluted with H2O (2.00 mL) and washed twice with DCM (2.00 mL x 2). The aqueous layer was acidified with 1N HCl at 0 °C until pH 1-2 and extracted with EtOAc (3.00 mL). The organic layer was washed with brine (3.00 mL), dried over MgSO4, filtered, and concentrated under reduced pressure. The residue was purified by column chromatography on SiO.sub.2 (10% MeOH in DCM) to give compound BnH12071 (35.0 mg, 44%) as a white solid.

[0472] 1H NMR (400 MHz, DMSO-d6) δ 9.16 (d, J = 12.7 Hz, 2H), 7.77 (d, J = 7.9 Hz, 2H), 7.38 (dd, J = 19.1, 7.8 Hz, 2H), 4.75 (s, 0.5H), 4.58 - 4.51 (d, J = 13.8 Hz, 0.5H), 4.29 (s, 0.5H), 3.92 (d, J = 13.8 Hz, 0.5H), 3.80 (tt, J = 15.5, 7.8 Hz, 2H), 3.05 (t, J = 12.8 Hz, 1H), 2.60 (t, J = 12.9 Hz, 1H), 2.24 - 2.11 (m, 1H), 1.86 - 1.77 (m, 1H), 1.66 (d, J = 13.3 Hz, 1H), 1.53 (q, J = 5.7, 4.6 Hz, 2H), 1.08 (dd, J = 16.6, 6.9 Hz, 3H).

[0473] Preparation Example 7. Preparation of active compounds according to representative reaction scheme 8

[0474] [Reaction Scheme 8]

[0475] JPEG2025512899000032.jpg65145

[0476] JPEG2025512899000033.jpg156125

[0477] (1) Synthesis of compound BnH12091

[0478] (3R,6S)-6-Methyl-1-(2-(3-(pyridin-4-yl)phenyl)acetyl)piperidine-3-carboxylic acid

[0479] Step 1: Methyl (3R,6S)-1-(2-(3-bromophenyl)acetyl)-6-methylpiperidine-3-carboxylate

[0480] To a solution of 3-bromophenylacetic acid (800 mg, 3.72 mmol) in DMF (12.4 mL) was added methyl (3R,6S)-rel-6-methyl-3-piperidinecarboxylate (614 mg, 3.91 mmol), DIPEA (0.972 mL, 5.58 mmol), HOBt (603 mg, 4.46 mmol), and EDC·HCl (692 mg, 4.46 mmol). The mixture was stirred at room temperature for 4 h. The mixture was diluted with EtOAc (15.0 mL) and washed with saturated aqueous NH4Cl (15.0 mL) and brine (15.0 mL). The organic layer was dried over MgSO4, filtered, and concentrated under reduced pressure. The residue was purified by column chromatography on SiO2 (n-hexane:EtOAc = 2:1 to 1:1) to give the desired compound (990 mg, 75%) as a colorless oil.

[0481] Step 2: Synthesis of methyl (3R,6S)-6-methyl-1-(2-(3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenyl)acetyl)piperidine-3-carboxylate (8-2)

[0482] A suspension of methyl (3R,6S)-1-(2-(3-bromophenyl)acetyl)-6-methylpiperidine-3-carboxylate (990 mg, 2.80 mmol), bis(pinacolato)diborane (501 mg, 1.97 mmol), and KOAc (823 mg, 3.07 mmol) in 1,4-dioxane (9.70 mL) was vigorously stirred and bubbled with N2 gas for 5 min, followed by the addition of Pd(dppf)Cl2 (205 mg, 0.279 mmol). The mixture was bubbled with N2 gas for 5 min and refluxed at 100 °C for 14 h. After completion of the reaction, the reaction mixture was filtered through a celite pad and washed with DCM (10.0 mL × 3). The combined organic layers were washed with H2O (30.0 mL), dried over MgSO4, filtered, and concentrated under reduced pressure. The residue was purified by column chromatography on SiO2 (n-hexane:EtOAc = 2:1 to 1:1) to give the desired compound (840 mg, 75%) as a yellowish oil.

[0483] Step 3: Synthesis of compound BnH12091

[0484] A solution of methyl (3R,6S)-6-methyl-1-(2-(3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenyl)acetyl)piperidine-3-carboxylate (8-2, 215 mg, 0.536 mmol), 4-bromopyridine hydrochloride (104 mg, 0.536 mmol), and Na2CO3 (170 mg, 1.61 mmol) in MeCN / H2O (2:1, 6.70 mL) was vigorously stirred and bubbled with N2 gas for 5 min. To the mixture was added Pd(dppf)Cl2 (31.0 mg, 0.042 mmol). The mixture was bubbled with N2 gas for 5 min and refluxed at 90 °C for 2 h. The reaction mixture was cooled to room temperature and 1N NaOH (2.14 mL, 2.14 mmol) was added. The mixture was stirred at room temperature for 2 h. The mixture was diluted with H2O (7.00 mL) and washed with EtOAc (7.00 mL x 3). 1N HCl was added to the aqueous layer to adjust the pH value to pH 4-5, and the mixture was extracted with EtOAc (7.00 mL x 3). The combined organic layers were dried over MgSO4, filtered, and concentrated under reduced pressure. Et2O (5.00 mL) was added to the residue and stirred at room temperature for 30 min. The precipitate was filtered and washed with Et2O (3.0 mL) to give compound BnH12091 (112 mg, 62%) as an off-white solid.

[0485] 1H NMR (400 MHz, DMSO-d6) δ 12.43 (s, 1H), 8.64-8.63 (d, J = 5.1 Hz, 2H), 7.69-7.63 (d, J = 7.3 Hz, 4H), 7.48-7.44 (t, J = 7.7 Hz, 1H), 7.35-7.29 (dd, J = 15.8, 7.6 Hz, 1H), 4.74 (s, 0.5H), 4.56-4.52 (m, 0.5H), 4.32 (s, 0.5H), 3.98-3.94 (d, J = 14.0 Hz, 0.5H), 3.89-3.77 (m, 2H), 3.07-3.01 (t, J = 12.9 Hz, 0.5H), 2.62-2.58 (d, J = 12.9 Hz, 0.5H), 2.18-2.14 (d, J = 14.5 Hz, 1H), 1.77 (s, 1H), 1.70-1.64 (m, 1H), 1.52 (s, 2H), 1.09-1.04 (m, 3H).

[0486] (2) Synthesis of compound BnH12092

[0487] (3R,6S)-1-(2-(3-(6-methoxypyridin-3-yl)phenyl)acetyl)-6-methylpiperidine-3-carboxylic acid

[0488] Step 1: Synthesis of compound BnH12092

[0489] A solution of methyl (3R,6S)-6-methyl-1-(2-(3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenyl)acetyl)piperidine-3-carboxylate (8-2, 225 mg, 0.561 mmol), 5-bromo-2-methoxypyridine (105 mg, 0.561 mmol), and Na2CO3 (119 mg, 1.12 mmol) in MeCN / H2O (2:1, 7.50 mL) was vigorously stirred and bubbled with N2 gas for 5 min. To the mixture was added Pd(dppf)Cl2 (41.0 mg, 0.056 mmol). The mixture was bubbled with N2 gas for 5 min and refluxed at 90 °C for 2 h. The reaction mixture was cooled to room temperature and 1 N NaOH (1.12 mL, 1.12 mmol) was added. The mixture was stirred at room temperature for 14 h. The mixture was diluted with H2O (7.00 mL) and washed with EtOAc (7.00 mL x 3). 1N HCl was added to the aqueous layer to adjust the pH value to pH 4-5, and the mixture was extracted with EtOAc (7.00 mL x 3). The combined organic layers were dried over MgSO4, filtered, and concentrated under reduced pressure. The residue was purified by column chromatography on SiO2 (DCM:MeOH = 20:1-10:1). Et2O (4.00 mL) was added to the residue and stirred at room temperature for 30 min. The precipitate was filtered and washed with Et2O (2.0 mL) to give compound BnH12092 (106 mg, 52%) as an off-white solid.

[0490] 1H NMR (400 MHz, DMSO-d6) δ 12.43 (s, 1H), 8.45 (dd, J = 7.3, 2.7 Hz, 1H), 7.97 (dd, J = 8.4, 2.5 Hz, 1H), 7.56 - 7.46 (m, 2H), 7.41 (t, J = 7.6 Hz, 1H), 7.22 (dd, J = 16.2, 7.6 Hz, 1H), 6.92 (d, J = 8.6 Hz, 1H), 4.75 (s, 0.5H), 4.59 - 4.50 (m, 0.5H), 4.32 (s, 0.5H), 3.90 (s, 3.5H), 3.87 - 3.72 (m, 2H), 3.03 (t, J = 12.9 Hz, 0.5H), 2.62 (t, J = 12.7 Hz, 0.5H), 2.17 (q, J = 10.2, 8.7 Hz, 1H), 1.79 (d, J = 12.0 Hz, 1H), 1.67 (dd, J = 12.5, 6.2 Hz, 1H), 1.51 (d, J = 12.4 Hz, 2H), 1.08 (dt, J = 12.6, 4.7 Hz, 3H).

[0491] (3) Synthesis of compound BnH12093

[0492] (3R,6S)-1-(2-(3-(5-fluoropyrimidin-2-yl)phenyl)acetyl)-6-methylpiperidine-3-carboxylic acid

[0493] Step 1: Synthesis of compound BnH12093

[0494] A solution of methyl (3R,6S)-6-methyl-1-(2-(3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenyl)acetyl)piperidine-3-carboxylate (8-2, 213 mg, 0.531 mmol), 2-bromo-5-fluoropyrimidine (93.9 mg, 0.531 mmol), and Na2CO3 (113 mg, 1.06 mmol) in MeCN / H2O (2:1, 6.84 mL) was vigorously stirred and bubbled with N2 gas for 5 min. To the mixture was added Pd(dppf)Cl2 (38.8 mg, 0.053 mmol). The mixture was bubbled with N2 gas for 5 min and refluxed at 90 °C for 2 h. The reaction mixture was cooled to room temperature and 1 N NaOH (1.12 mL, 1.12 mmol) was added. The mixture was stirred at room temperature for 17 h. The mixture was diluted with H2O (7.00 mL) and washed with EtOAc (7.00 mL x 3). 1N HCl was added to the aqueous layer to adjust the pH value to pH 3-4, and the mixture was extracted with EtOAc (7.00 mL x 3). The combined organic layers were dried over MgSO4, filtered, and concentrated under reduced pressure. The residue was purified by column chromatography on SiO2 (3-5% MeOH in DCM). Et2O (5.00 mL) was added to the residue and stirred at room temperature for 30 min. The precipitate was filtered and washed with Et2O (3.0 mL) to give compound BnH12093 (122 mg, 64%) as a beige solid.

[0495] 1H NMR (400 MHz, DMSO-d6) δ 12.39 (s, 1H), 8.98 (s, 2H), 8.22 (t, J = 10.5 Hz, 2H), 7.53 - 7.34 (m, 2H), 4.76 (s, 0.5H), 4.55 (dd, J = 13.2, 4.0 Hz, 0.5H), 4.37 - 4.22 (m, 0.5H), 3.98 - 3.76 (m, 2.5H), 3.03 (t, J = 12.8 Hz, 0.5H), 2.62 (t, J = 12.7 Hz, 0.5H), 2.18 (q, J = 10.8, 9.2 Hz, 1H), 1.79 (d, J = 13.0 Hz, 1H), 1.65 (td, J = 12.5, 7.6 Hz, 1H), 1.51 (d, J = 9.5 Hz, 2H), 1.08 (dt, J = 7.1, 4.4 Hz, 3H).

[0496] Preparation Example 8. Preparation of active compounds by methods other than the representative reaction scheme

[0497] (1) Synthesis of BnH12002

[0498] (3R,6S)-1-(2-(4'-fluoro-[1,1'-biphenyl]-4-yl)acetyl)-6-methylpiperidine-3-carboxylic acid

[0499] JPEG2025512899000034.jpg71151

[0500] Step 1: Compound 1 (293 mg, 1.27 mmol) was dissolved in 5 mL of DCM and oxalyl chloride (163 μL, 1.91 mmol) was added dropwise at 0° C. for 5 min, followed by the addition of DMF (1 drop) with catalyst. After stirring at room temperature for 6 h, the mixture was concentrated under reduced pressure to give 316 mg of compound 2. The reaction was carried forward to the next step without further purification.

[0501] Step 2: Compound 3 (200 mg, 1.27 mmol) was dissolved in 10 mL of DCM, and triethylamine (354 μL, 2.54 mmol) was added at 0° C. Compound 2 (316 mg, 1.27 mmol) was dissolved in 5 mL of DCM, and added dropwise at 0° C., followed by stirring at room temperature for 2 hours. The reaction mixture was worked up with DCM and water, and then used as a column to obtain 213 mg of compound 4 (yield 45%).

[0502] Step 3: Compound 4 (100 mg, 473 μmol) was dissolved in 5 mL of THF / methanol (1:1), 1N-NaOH (947 μL, 947 μmol) was added, and the mixture was stirred at room temperature for 12 hours. After the reaction was completed, the pH was adjusted to 3 using 1N-HCl, extracted with ethyl acetate, and concentrated under reduced pressure. 75 mg of the target compound 5 was obtained from the resulting reaction mixture using prep TLC (yield 45%).

[0503] The identity was confirmed by NMR spectrum: 1H NMR (CDCl3, 400 MHz): δ 7.22 (1H, m), 7.08 (1H, m), 6.91 (1H, bs), 4.75(1H, m), 3.86 (1H, m), 3.71(2H, m), 3.01 (1H, m), 2.28 (1H, m), 1.91 (1H, bs), 1.74(1H, m), 1.58 (1H, bs), 1.13(3H, m).

[0504] (2) Synthesis of BnH12003

[0505] (3R,6S)-6-Methyl-1-(2-(4-(pyrimidin-5-yl)phenyl)acetyl)piperidine-3-carboxylic acid

[0506] JPEG2025512899000035.jpg107151

[0507] Step 1: Compound 1 (547 mg, 2.54 mmol) was dissolved in 10 mL of DCM and oxalyl chloride (327 μL, 3.81 mmol) was added dropwise at 0° C. for 5 min, followed by DMF (1 drop) with catalyst. After stirring at room temperature for 6 h, the mixture was concentrated under reduced pressure to give 593 mg of compound 2, which was carried forward to the next step without further purification.

[0508] Step 2: Compound 3 (400 mg, 2.54 mmol) was dissolved in 10 mL of DCM, and triethylamine (708 μL, 5.08 mmol) was added at 0° C. Compound 2 (593 mg, 2.54 mmol) was dissolved in 5 mL of DCM, and added dropwise at 0° C., followed by stirring at room temperature for 2 hours. The reaction mixture was worked up with DCM and water, and then used as a column to obtain 347 mg of compound 4 (yield 38%).

[0509] Step 3: Compound 4 (300 mg, 846 μmol), compound 5 (104 mg, 846 μmol), Pd(PPh3)4 (49 mg, 43 μmol), and K2CO3 (234 mg, 1.69 mmol) were dissolved in toluene (3 mL), ethanol (1 mL), and H2O (1 mL), degassed under nitrogen, and refluxed for 12 h. After the reaction was completed, the mixture was extracted with EA and concentrated under reduced pressure to give 38 mg of compound 6 (13% yield) using prep TLC.

[0510] Step 4: Compound 6 (38 mg, 107 μmol) was dissolved in 3 mL of THF / methanol (1:1), 1N-NaOH (430 μL, 430 μmol) was added, and the mixture was stirred at room temperature for 12 hours. After the reaction was completed, the pH was adjusted to 3 using 1N-HCl, extracted with ethyl acetate, and then concentrated under reduced pressure. 12 mg of the target compound 7 was obtained from the resulting reaction mixture using prep TLC (yield 45%).

[0511] The identity was confirmed by NMR spectrum: 1H NMR (CDCl3, 400 MHz): δ 9.20 (1H, s), 8.97 (2H, m), 7.55 (2H, d, J=8Hz), 7.42 (2H, d, J=8Hz), 4.88(1H, m), 4.15 (1H, m), 3.82(2H, s), 3.01 (1H, m), 2.28 (1H, m), 1.98 (1H, m), 1.80(1H, m), 1.60 (2H, m), 1.17(3H, m).

[0512] (3) Synthesis of BnH12057

[0513] (3R,6S)-6-Methyl-1-(2-(4-(oxazol-5-yl)phenyl)acetyl)piperidine-3-carboxylic acid

[0514] JPEG2025512899000036.jpg20145

[0515] Step 1: Methyl (3R,6S)-6-methyl-1-(2-(4-(oxazol-5-yl)phenyl)acetyl)piperidine-3-carboxylate (g)

[0516] A solution of compound 1-2-BnH12032 (500 mg, 1.41 mmol), oxazole (97.5 mg, 1.41 mmol), and Bu4NOAc (553 mg, 1.84 mmol) in DMA (4.71 mL) was vigorously stirred, bubbled with N2 gas for 5 min, and Pd(Oac)2 (95.1 mg, 0.423 mmol) was added. The mixture was bubbled with N2 gas for 5 min and refluxed at 70 °C for 17 h. The mixture was then cooled to room temperature and concentrated. The residue was purified by column chromatography on SiO2 (n-hexane: EtOAc = 1:1) to give the desired compound (110 mg, 22%) as a brown oil.

[0517] Step 2: Synthesis of compound BnH12057

[0518] To a solution of methyl (3R,6S)-6-methyl-1-(2-(4-(oxazol-5-yl)phenyl)acetyl)piperidine-3-carboxylate (25.0 mg, 0.073 mmol) in THF (0.252 mL) was added 1N NaOH (0.110 mL, 0.110 mmol). The mixture was stirred at room temperature for 2 h. The mixture was diluted with H2O (2.00 mL) and washed with DCM (2.00 × 2). The aqueous layer was acidified with 1N HCl at 0 °C until pH 1-2 and extracted with EtOAc (3.00 mL). The organic layer was washed with brine (3.00 mL), dried over MgSO4, filtered, and concentrated under reduced pressure to provide compound BnH12057 (10.0 mg, 41%) as a white solid.

[0519] 1H NMR (400 MHz, Chloroform-d) δ 7.93 (s, 1H), 7.62 (d, J = 8.1 Hz, 2H), 7.37 - 7.29 (m, 3H), 4.98 (s, 0.5H), 4.83 (d, J = 13.9 Hz, 0.5H), 4.15 (d, J = 16.6 Hz, 0.5H), 3.92 (d, J = 13.9 Hz, 0.5H), 3.78 (s, 2H), 3.16 (t, J = 12.9 Hz, 0.5H), 2.79 (t, J = 12.9 Hz, 0.5H), 2.40 (s, 0.5H), 2.16 (d, J = 7.3 Hz, 0.5H), 1.94 (d, J = 14.3 Hz, 1H), 1.80 (q, J = 12.2, 11.3 Hz, 1H), 1.67 - 1.44 (m, 2H), 1.14 (d, J = 6.9 Hz, 3H).

[0520] (4) Synthesis of compound BnH12067 (anhydrous form)

[0521] (3S,6R)-6-Methyl-1-(2-(4-(pyrazin-2-yl)phenyl)acetyl)piperidine-3-carboxylic acid

[0522] JPEG2025512899000037.jpg65140

[0523] Step 1: Sodium 2-(4-(pyrazin-2-yl)phenyl)acetate (BnH12067-2)

[0524] A solution of 2-(4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenyl)acetic acid (6-1-BnH12064-1, 100 mg, 0.382 mmol), 2-bromopyrazine (66.3 mg, 0.420 mmol), and Na2CO3 (121 mg, 1.15 mmol) in MeCN / HO (2:1, 4.76 mL) was vigorously stirred and bubbled with N2 gas for 5 min, and Pd(dppf)Cl2 (27.9 mg, 0.038 mmol) was added. The mixture was bubbled with N2 gas for 5 min and refluxed at 90 °C for 3 h. Subsequently, the mixture was cooled to room temperature. The mixture was diluted with HO (10.0 mL) and washed with DCM (5.00 mL × 3). The aqueous layer was concentrated under reduced pressure to give the desired compound (90.0 mg, quant.) as a yellow solid, which was used without further purification.

[0525] Step 2: Methyl (3S,6R)-6-methyl-1-(2-(4-(pyrazin-2-yl)phenyl)acetyl)piperidine-3-carboxylate (BnH12067-1)

[0526] To a solution of sodium 2-(4-(pyrazin-2-yl)phenyl)acetate (BnH12067-2, 90 mg, 0.381 mmol) in DMF (1.27 mL) was added methyl (3R,6S)-rel-6-methyl-3-piperidinecarboxylate (anhydrous form) (62.9 mg, 0.400 mmol), DIPEA (0.100 mL, 0.572 mmol), HOBt (61.8 mg, 0.457 mmol), and EDC·HCl (71.0 mg, 0.457 mmol). The mixture was stirred at room temperature for 6 h. The mixture was diluted with EtOAc (2.00 mL) and washed with saturated aqueous NH4Cl (2.00 mL) and brine (2.00 mL). The organic layer was dried over MgSO4, filtered, and concentrated under reduced pressure. The residue was purified by column chromatography on SiO.sub.2 (5% MeOH in DCM) to give the desired compound (67.0 mg, 50%) as a brown oil.

[0527] Step 3: Synthesis of compound BnH12067

[0528] To a solution of methyl (3S,6R)-6-methyl-1-(2-(4-(pyrazin-2-yl)phenyl)acetyl)piperidine-3-carboxylate (67.0 mg, 0.190 mmol) in THF (1.00 mL) was added 1N NaOH (0.284 mL, 0.284 mmol). The mixture was stirred at room temperature for 2 h. The mixture was diluted with H2O (2.00 mL) and washed twice with DCM (2.00 mL x 2). The aqueous layer was acidified with 1N HCl at 0 °C until pH 1-2 and extracted with EtOAc (2.00 mL). The organic layer was washed with brine (3.00 mL), dried over MgSO4, filtered, and concentrated under reduced pressure to provide compound BnH12067 (49.9 mg, 88%) as an off-white solid.

[0529] 1H NMR (400 MHz, DMSO-d6) δ 9.25 (s, 1H), 8.71 (t, J = 2.0 Hz, 1H), 8.60 (d, J = 2.5 Hz, 1H), 8.10 (d, J = 7.9 Hz, 2H), 7.39 (dd, J = 18.5, 7.9 Hz, 2H), 4.75 (s, 0.5H), 4.55 (d, J = 13.7 Hz, 0.5H), 4.28 (s, 0.5H), 3.92 (d, J = 13.8 Hz, 0.5H), 3.88 - 3.73 (m, 2H), 3.04 (t, J = 12.9 Hz, 0.5H), 2.62 (t, J = 12.8 Hz, 0.5H), 2.19 (ddt, J = 11.7, 8.0, 4.0 Hz, 1H), 1.79 (d, J = 11.9 Hz, 1H), 1.66 (d, J = 13.7 Hz, 1H), 1.52 (s, 2H), 1.07 (dd, J = 9.2, 7.0 Hz, 3H).

[0530] (5) Synthesis of compound BnH12095

[0531] (3R,6S)-6-メチル-1-(2-(3-(ピリミジン-5-イル)フェニル)アセチル)ピペリジン-3-カルボン acid

[0532] JPEG2025512899000038.jpg57142

[0533] Stage 1: Synthesis of compound BnH12095

[0534] A solution of methyl (3R,6S)-1-(2-(3-bromophenyl)acetyl)-6-methylpiperidine-3-carboxylate (8-1, 170 mg, 0.424 mmol), 5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)pyrimidine (87.3 mg, 0.424 mmol), and Na2CO3 (89.8 mg, 0.847 mmol) in MeCN / H2O (2:1, 5.46 mL) was vigorously stirred and bubbled with N2 gas for 5 min. To the mixture was added Pd(dppf)Cl2 (31.0 mg, 0.042 mmol). The mixture was bubbled with N2 gas for 5 min and refluxed at 90 °C for 3 h. The reaction mixture was cooled to room temperature and 1 N NaOH (0.847 mL, 0.847 mmol) was added. The mixture was stirred at room temperature for 17 h. The mixture was diluted with H2O (5.00 mL) and washed with EtOAc (5.00 mL x 3). 1N HCl was added to the aqueous layer to adjust the pH value to pH 2-3, and the mixture was extracted with EtOAc (5.00 mL x 3). The combined organic layers were dried over MgSO4, filtered, and concentrated under reduced pressure. The residue was purified by column chromatography on SiO2 (3-5% MeOH in DCM) to give compound BnH12095 (95.0 mg, 66%) as an off-white solid.

[0535] 1H NMR (400 MHz, DMSO-d6) δ 12.40 (s, 1H), 9.20 (s, 1H), 9.12 (d, J = 8.6 Hz, 2H), 7.68 (d, J = 9.7 Hz, 2H), 7.49 (t, J = 7.7 Hz, 1H), 7.33 (dd, J = 15.5, 7.7 Hz, 1H), 4.74 (s, 0.5H), 4.61 - 4.49 d, J = 13.6 Hz, 0.5H), 4.33 (s, 0.5H), 3.98 (d, J = 13.6 Hz, 0.5H), 3.91 - 3.74 (m, 2H), 3.06 (t, J = 12.9 Hz, 0.5H), 2.62 (t, J = 12.7 Hz, 0.5H), 2.15 (dd, J = 25.2, 12.4 Hz, 1H), 1.76 (s, 1H), 1.66 (dq, J = 12.1, 6.6 Hz, 1H), 1.54 (d, J = 9.8 Hz, 2H), 1.14 - 1.03 (m, 3H).

[0536] (6) Synthesis of compound BnH12097

[0537] 6-Ethyl-1-(2-(4-(pyrimidin-5-yl)phenyl)acetyl)piperidine-3-carboxylic acid

[0538] JPEG2025512899000039.jpg32143

[0539] Step 1: Ethyl 1-(2-(4-bromophenyl)acetyl)-6-ethylpiperidine-3-carboxylate (BnH12097-1)

[0540] To a solution of 2-(4-bromophenyl)acetic acid (700 mg, 3.26 mmol) in DMF (9.00 mL) was added ethyl 6-ethylpiperidine-3-carboxylate (p, 633 mg, 3.42 mmol), DIPEA (1.70 mL, 9.77 mmol), HOBt (528 mg, 3.91 mmol), and EDC·HCl (749 mg, 3.91 mmol). The mixture was stirred at room temperature for 17 h. The reaction mixture was diluted with EtOAc (25.0 mL) and washed with saturated aqueous NH4Cl (25.0 mL) and brine (25.0 mL). The organic layer was dried over Na2SO4, filtered, and concentrated under reduced pressure. The residue was purified by column chromatography on SiO2 (n-hexane:EtOAc = 1:0 to 2:1) to give the desired compound (302 mg, 36%, 2 steps) as a yellow oil.

[0541] Step 2: Synthesis of compound BnH12097

[0542] A solution of ethyl 1-(2-(4-bromophenyl)acetyl)-6-ethylpiperidine-3-carboxylate (150 mg, 0.392 mmol), 5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)pyrimidine (80.9 mg, 0.392 mmol), and Na2CO3 (83.2 mg, 0.785 mmol) in MeCN:H2O (2:1, 4.80 mL) was vigorously stirred, bubbled with N2 gas for 5 min, and Pd(dppf)Cl2 (28.7 mg, 0.039 mmol) was added. The mixture was bubbled with N2 gas for 5 min and refluxed at 90 °C for 3 h. After stirring, the mixture was cooled to room temperature and 1N NaOH (0.970 mL, 0.970 mmol) was added and stirred at 50 °C for 17 h. The reaction mixture was diluted with H2O (5.00 mL) and washed twice with DCM (5.00 mL x 2). The aqueous layer was acidified with 1N HCl at 0 °C until pH 1-2 and extracted with EtOAc (5.00 mL). The organic layer was dried over Na2SO4, filtered, and concentrated under reduced pressure. The residue was purified by column chromatography on SiO2 (DCM:MeOH = 1:0-4:1) to give compound BnH12097 (35.3 mg, 26%) as a light brown solid.

[0543] 1H NMR (400 MHz, DMSO-d6) δ 9.18 - 9.14 (m, 3H), 7.77 (d, J = 7.8 Hz, 2H), 7.39 (dd, J = 14.3, 7.9 Hz, 2H), 4.58 - 4.54 (m, 1H), 3.99 - 3.71 (m, 3.5H), 2.96 (t, J = 12.9 Hz, 0.5H), 2.19 - 2.09 (m, 1H), 1.79 (s, 1H), 1.65 - 1.39 (m, 5H), 0.77 (dt, J = 14.0, 7.3 Hz, 3H).

[0544] (7) Synthesis of compound BnH12098

[0545] 6-Isopropyl-1-(2-(4-(pyrimidin-5-yl)phenyl)acetyl)piperidine-3-carboxylic acid

[0546] JPEG2025512899000040.jpg31143

[0547] Step 1: Synthesis of ethyl 1-(2-(4-bromophenyl)acetyl)-6-isopropylpiperidine-3-carboxylate (BnH12098-1)

[0548] To a solution of 2-(4-bromophenyl)acetic acid (620 mg, 2.91 mmol) in DMF (9.69 mL) was added ethyl 6-isopropylpiperidine-3-carboxylate (s, 608 mg, 3.05 mmol), DIPEA (1.52 mL, 8.72 mmol), HOBt (471 mg, 3.49 mmol), and EDC·HCl (542 mg, 3.49 mmol). The mixture was stirred at room temperature for 17 h. The reaction mixture was diluted with EtOAc (20.0 mL) and washed with saturated aqueous NH4Cl (20.0 mL) and brine (20.0 mL). The organic layer was dried over MgSO4, filtered, and concentrated under reduced pressure. The residue was purified by column chromatography on SiO2 (n-hexane:EtOAc = 3:1 to 2:1) to give the desired compound (618 mg, 54%, 2 steps) as a yellow oil.

[0549] Step 2: Synthesis of compound BnH12098

[0550] A solution of ethyl 1-(2-(4-bromophenyl)acetyl)-6-isopropylpiperidine-3-carboxylate (300 mg, 0.748 mmol), 5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)pyrimidine (154 mg, 0.748 mmol), and Na2CO3 (158 mg, 1.50 mmol) in MeCN:H2O (2:1, 9.64 mL) was vigorously stirred, bubbled with N2 gas for 5 min, and Pd(dppf)Cl2 (54.7 mg, 0.075 mmol) was added. The mixture was bubbled with N2 gas for 5 min and refluxed at 90 °C for 2 h. After stirring, the mixture was cooled to room temperature and 1N NaOH (1.50 mL, 1.50 mmol) was added and stirred at 50 °C for 17 h. The reaction mixture was diluted with H2O (5.00 mL) and washed twice with DCM (5.00 mL x 2). The aqueous layer was acidified with 1N HCl at 0 °C until pH 1-2 and extracted with EtOAc (10.0 mL). The organic layer was dried over MgSO4, filtered, and concentrated under reduced pressure. The residue was purified by column chromatography on SiO2 (5-10% MeOH in DCM) to give compound BnH12098 (90 mg, 33%) as an off-white solid.

[0551] 1H NMR (400 MHz, DMSO-d6) δ 12.39 (s, 1H), 9.20 - 9.12 (m, 3H), 7.78 (d, J = 7.9 Hz, 2H), 7.40 (dd, J = 10.7, 7.9 Hz, 2H), 4.62 - 4.54 (m, 0.5H), 4.25 - 4.13 (m, 0.5H), 3.97 (dd, J = 14.9, 4.1 Hz, 0.5H), 3.93 - 3.68 (m, 2H), 3.60 (d, J = 10.2 Hz, 0.5H), 2.96 (t, J = 13.0Hz, 1H), 2.29 - 1.96 (m, 2H), 1.82 (d, J = 14.5 Hz, 1.5H), 1.70 - 1.55 (m, 1H), 1.32 (dt, J = 28.1, 13.4 Hz, 1H), 0.92 (dd, J = 17.3, 6.5 Hz, 3H), 0.75 (dd, J = 25.8, 6.6 Hz, 3H).

[0552] The compounds obtained in the above Preparation Examples were subjected to biological assays as follows.

[0553] Working Example

[0554] Example 1. Cell Culture and Maintenance

[0555] Human anaplastic neuroblastoma cell line SH-SY5Y and human embryonic kidney cell line (HEK)-293T were purchased from the Korean Cell Line Bank (Korea) and Takara (Japan), respectively. Cells were grown in HyClone Minimal Essential Medium (MEM) and Dulbecco's Modified Eagle Medium (DMEM), respectively, supplemented with 10% (v / v) fetal bovine serum (FBS), penicillin (100 U / mL), and streptomycin (100 mg / mL). Cells were maintained at 37°C in a humidified atmosphere of 95% air and 5% CO2. Cells were passaged up to 10 times. Cells were detached using 0.05% trypsin-EDTA solution and seeded into new culture dishes for maintenance.

[0556] Meanwhile, primary cortical neuron cells were prepared by dissecting the cerebral cortex from mouse fetuses on day 18 of gestation, incubating the cells for 30 minutes in prep media (composition (mM): NaCl 107.5, NaH2PO4 1, NaHCO3 3.5, glucose 11, HEPS 1%, penicillin-streptomycin 1%) containing 0.25% trypsin and DNase1dl, and then dissociating and seeding the cells into single cells. Primary cortical neuron cells were grown in Neurobasal medium supplemented with 1% (v / v) B27, 1% penicillin (100 U / mL), streptomycin (100 mg / mL), and 1% Glutamax. The cells were maintained at 37°C in a humidified atmosphere of 95% air and 5% CO2. The primary cortical neuron cells were cultured for 14 days and then evaluated for drug efficacy.

[0557] Example 2. Cytotoxicity test

[0558] To investigate the cytotoxicity of the compounds described herein in human cell lines, SH-SY5Y and 293T cells were used. Approximately 8,000 cells (for 293T) and 15,000 cells (for SH-SY5Y) were plated per well of a 96-well plate (SPL Life Sciences Co., Ltd., Pocheon, Korea). After 24 hours of incubation at 37°C in a humidified atmosphere containing 5% CO2, the culture medium was replaced with a series of concentrations of the test compound diluted in the culture medium. The co-incubation time was 24 hours, and six replicates were performed for each measurement. Finally, 10 μL of CCK-8 reagent (Dojindo Laboratories, Rockville, MD, USA) was added to each well, and after 3 minutes of incubation at 37°C, the OD at 450 nm was measured using a multifunctional microplate reader (Tecan Infinite M200 Pro). These measurements were compared with those of the control group to calculate cell viability, which was expressed as a percentage of the control group. The IC was calculated using SPSS software. 50 Values ​​were calculated. To do this, the absorbance of blank wells containing growth medium only was subtracted from the values ​​for wells containing cells. The background (vehicle) reading was set to 100% viability, and the viability of each test solution was then calculated.

[0559] Cell viability (%)=[(As-Ab) / (Ac-Ab)]×100

[0560] Data are presented as mean and standard error of the mean (SEM). Statistical analysis was performed by one-way ANOVA with Tukey's post-hoc test (GraphPad Prism 5 software package, version 5.02, GraphPad Software Inc., USA). Significance was accepted at p < 0.05.

[0561] Example 3. Measurement of expression levels of BDNF and / or GluN2B

[0562] Example 3.1. RT-PCR

[0563] RT-PCR was used to measure the amount of BDNF and GluN2B in response to the compounds of the present invention. 293T cells were isolated using 0.05% trypsin-EDTA solution and seeded into 24-well or 6-well plates for RNA work. After 24 hours of incubation at 37°C in a humidified atmosphere containing 5% CO2, the culture medium was replaced with medium containing a series of concentrations of the test compound diluted in DMEM without FBS. A time course experiment was performed. The same amount of DMSO was used as a negative control.

[0564] After compound treatment, RNA was isolated from the cells. After removing the growth medium, 1 × 10 5 ~10 7 0.3-0.4 mL of TRIzol per cell TM Cells were lysed by adding 1 mL of TRIzol directly to the culture dish. After incubation at 4 °C for 10 min, the samples were centrifuged at 12,000 × g for 10 min at 4 °C. The total RNA precipitate formed a white gel-like pellet at the bottom of the tube. After discarding the supernatant, the pellet was lysed in 1 mL of TRIzol, which was used for lysis. TM The pellet was resuspended in 1 mL of 75% ethanol per well. After brief vortexing, the samples were centrifuged at 7500 x g for 5 min at 4°C. The supernatant was discarded with a micropipette and vacuum or air dried for 5–10 min. The pellet was resuspended in 20–50 μL of RNase-free water. RNA concentration was determined using Thermo Scientific TM μDrop TM The A230 / 260 and A260 / 280 ratios were measured to determine RNA quality.

[0565] On ice, experimental RNA (up to 1 μg in 4 ul) and 1 ul (100 pmol) of oligo dT primer were brought up to a final volume of 5 μl per RT reaction in nuclease-free water. After tightly closing each RNA tube, the tubes were placed (in the PCR machine) at 70°C for 5 minutes. After removing the tubes, the tubes were immediately cooled in ice-water for at least 5 minutes. Each tube was centrifuged for 10 seconds in a microcentrifuge to collect condensate and maintain the original volume. The tubes were kept closed on ice until the reverse transcription reaction mixture was added. The reverse transcription reaction mixture was prepared as follows:

[0566] JPEG2025512899000041.jpg100145

[0567] Reverse transcription was performed using Improm-II Reverse Transcriptase (A3803) (Promega) and Recombinant RNasin Ribonuclease Inhibitor (N2515) (Promega) according to the manufacturer's guidelines. A 15 μl aliquot of the reverse transcription reaction mixture was added to each reaction tube on ice and the tubes were placed back into the PCR machine. PCR conditions were as follows:

[0568] 25°C, 5 min (annealing) → 42°C, 60 min (extension) → 70°C, 15 min (RT inactivation) → hold at 4°C

[0569] PCR products were analyzed by agarose gel electrophoresis. A housekeeping gene (ACTIN) was used as a PCR control. Replicate samples confirmed the validity of the changes observed at the RNA level. The primer sequences used were as follows:

[0570] JPEG2025512899000042.jpg79145

[0571] Example 3.2. Western Blotting

[0572] Western blot was used to measure the amount of GluN2B reacted to the compound of the present invention. Fetal cerebral cortex was excised from mice on day 17 of pregnancy, dissociated into single cells, cultured for 13 days, and treated with BnH drugs for 24 hours. Protein lysates were prepared from the drug-treated cells using RIPA protein lysis buffer containing protease and phosphate inhibitors. Protein concentration was determined using a BCA protein assay kit (Pierce Biotechnology) with bovine serum albumin as the standard. The same amount of protein (20 μg per lane) was separated by electrophoresis on SDS-polyacrylamide gel at 90 V for 2 h, and then the SDS-polyacrylamide gel was transferred to a PVDF membrane at 100 V for 1 h. The primary reaction was performed with GluN2B antibody (ab65783, Abcam) at 4 °C for 1 day, washed three times with 1X PBT at 10 min intervals, and then the secondary reaction was performed with anti-mouse antibody (sc-2357, Santa Cruz) for 1 h, and then washed three times with 1X PBT at 10 min intervals again. Western blotting was performed as described above to confirm GluN2B protein expression. Then, each blot was re-probed using anti-β-actin antibody (sc-47778, Santa Cruz). The housekeeping protein β-actin was detected as a normalization control.

[0573] As a result, changes in GluN2B protein were confirmed with various BnH12 series compounds, as shown in Table 6.

[0574] Data analysis

[0575] The vehicle was set to 1, and the increase or decrease in the BnH12 series was confirmed (N per group was 3 or more).

[0576] Example 4. Synaptic function test

[0577] Example 4.1. Os pump implantation

[0578] Eight-week-old male C57BL / 6J mice were anesthetized with isoflurane during surgery. Mice were then placed in a stereotaxic frame (RWD Life Science, China). The parietal and frontal bones were exposed by a thalamotomy of the scalp from the midline from eye level to the occipital prominence. After the skull was exposed, a small burr hole was drilled. An infusion cannula was placed at the following coordinates: (mouse: vertex -3.30, lateral -1.50, and ventral 0.7 mm). Sterile saline solution (+ / - compound of the invention) was perfused at a constant flow rate of 0.5 μL / h through the infusion cannula using a mini-Os pump (ALZET Osmotic Pumps, DURECT Corporation, CA, USA). Implantation was performed in layer 4 of the eastern barrel cortex. To estimate the extent of diffusion of the infused compound in layer 4 of the barrel cortex during pump infusion, methylene blue was first injected to stain the diffusion area. On average, methylene blue diffused up to 2000.0 μm from the injection site (total area of ​​barrel cortex region is approximately 1500 μm). Based on this analysis, barrel cortex area was recorded.

[0579] Example 4.2. Oral Administration of Compounds

[0580] BnH12003 was dissolved using DMSO:PEG400:saline (5:55:40) and administered at a dose of 25 mg / kg (200 μl / animal). Control groups were administered DMSO:PEG400:saline (5:55:40) (200 μl / animal). Compounds were administered once a day for 22 doses from day 10 to day 31, starting at 8 weeks of age.

[0581] Example 4.3. Intraperitoneal injection of compounds

[0582] Compounds were diluted to 25 mg / kg using DMSO:PEG400:saline (5:55:40). Compounds were administered intraperitoneally (intraperitoneal volume is 200 μl) at 10 am every day for about 11-12 days. Control groups were administered by intraperitoneal injection in the same manner using vehicle (DMSO:PEG400:saline (5:55:40)). Five mice were used for each compound and control group.

[0583] Example 4.4. Thalamocortical (TC) or hippocampal slice preparation

[0584] For preparation of TC or hippocampal slices, 9-10 week old male C57BL / 6J mice were anesthetized with isoflurane. Subsequently, the brains were rapidly cooled by light cardioplegia perfusion with ice-cold sucrose artificial cerebrospinal fluid (aCSF). Brains were isolated and placed in ice-cold sucrose aCSF. Paracoronal slices were prepared at a 50° angle to the midline with a 10° angle ramp. Slices were subsequently incubated in aCSF for 30 min at 35°C to recover. Slices were subsequently incubated in aCSF for 1-4 h at room temperature (23-25°C) before being placed in the experimental recording chamber. Standard aCSF was included saturated with 95% O2 / 5% CO2. Mouse brains were isolated and sectioned at 400 um at a 50° angle (55° angle for mice) and attached to a vibratome disc so that the sectioned part was at the bottom. Brain slices were incubated in aCSF solution for 30 min at 35°C or incubated in aCSF solution for 1 h at room temperature.

[0585] Composition of sucrose-based aCSF

[0586] JPEG2025512899000043.jpg171145

[0587] Composition of recording aCSF (room temperature: RT)

[0588] JPEG2025512899000044.jpg171145

[0589] Example 4.5. Electrophysiological experiments on barrel cortex

[0590] All electrophysiological experiments were performed at 27–29 °C. For electrophysiological experiments, electrodes with resistances of 3.5–4.2 mΩ were used, and stimulation was delivered to the ventral posteromedial nucleus (VPM) using concentric bipolar electrodes (FHC, Bowdoin, ME, USA). Under 4× magnification and differential interference contrast (DIC) optics, the somatosensory cortex was identified by the presence of a barrel and by the ability to evoke brief, constant field excitatory postsynaptic potentials (fEPSPs) upon VPM stimulation. Whole-cell voltage clamp recordings were performed in stellate cells (SC) in layer IV of the somatosensory barrel cortex using infrared illumination and differential interference contrast (DIC) optics. The composition of the whole-cell recording solution is presented as follows:

[0591] Whole Cell Recording Internal Solution

[0592] JPEG2025512899000045.jpg151145

[0593] Whole-cell recordings were made using a Multiclamp 700A (Molecular devices, Sunnyvale, CA, USA) digitized at 10 kHz and filtered at 2 kHz. During recordings, input and series resistances were monitored as previously described (Isaac, Crair, Nicoll, & Malenka, 1997, Silent synapses during development of thalamocortical inputs, DOI: 10.1016 / s0896-6273(00)80267-6). TC EPSCs were evoked by VB stimulation at 0.1 Hz and were recognized as monosynaptic if they showed a short and constant latency that did not change with increasing stimulation intensity, as previously described (Feldman, Nicoll, Malenka, & Isaac, 1998, Synaptic plasticity at thalamocortical synapses in developing rat somatosensory cortex: LTP, LTD, and silent synapses, https: / / doi.org / 10.1002 / (SICI)1097-4695(199910)41:1<92::AID-NEU12>3.0.CO;2-U; Isaac et al., 1997, Silent Synapses during Development of Thalamocortical Inputs, https: / / doi.org / 10.1016 / S0896-6273(00)80267-6).

[0594] For the minimum stimulation protocol (potency), the VB thalamus stimulation intensity was adjusted to determine the lowest intensity that evoked a mixture of synaptic responses and failures. The failure rate was calculated as the number of failures / total number of attempts. Potency was calculated as the mean EPSC peak amplitude excluding failures (Chittajallu & Isaac, 2010, Emergence of cortical inhibition by coordinated sensory-driven plasticity at distinct synaptic loci, DOI: 10.1038 / nn.2639; Isaac et al., 1997, Silent Synapses during Development of Thalamocortical Inputs, https: / / doi.org / 10.1016 / S0896-6273(00)80267-6; Stevens & Wang, 1995, Facilitation and depression at single central synapses, DOI: 10.1016 / 0896-6273(95)90223-6).

[0595] The criteria for single axon stimulation were all or no synaptic events and no change in the mean amplitude of EPSCs evoked by small increases in stimulation intensity, as previously reported (Chittajallu and Isaac, 2010, Emergence of cortical inhibition by coordinated sensory-driven plasticity at distinct synaptic loci, DOI: 10.1038 / nn.2639; Gil et al., 1999, Efficacy of Thalamocortical and Intracortical Synaptic Connections: Quanta, Innervation, and Reliability, https: / / doi.org / 10.1016 / S0896-6273(00)80788-6). Measurement parameters were as follows:

[0596] Measurement parameters

[0597] JPEG2025512899000046.jpg44146

[0598] Excitatory postsynaptic potentials (EPSCs) were recorded by whole-cell patch clamp technique using a patch clamp amplifier (Molecular Devices, USA). The measurement electrodes were borosilicate glass capillaries with resistances of 3.5 MΩ to 4.2 MΩ when filled with the electrode's internal solution. The brain slices in the chambers were placed on a vertical microscope, fixed with anchors, and the recording aCSF solution was allowed to flow at a rate of 5 to 10 mL / min by gravity. The stimulator electrode was placed at the junction between the thalamic fibers and the S1 barrel cortex (layer 4) region, and cells in the region were located and recorded. After whole-cell patching, the membrane voltage was fixed at -70 mV (to measure AMPA current), and EPSCs were confirmed by applying stimulation, and the stimulator was manipulated to determine the minimum stimulation intensity at which the current on / off was 50%. The amplitude was measured for 3 minutes 20 seconds or 8 minutes 20 seconds of recording and tracing for the current (total of 20 or 50 traces). For potency, EPSC size was set to 50–100 pA at -70 mV and five traces were measured at 0 mV.

[0599] Example 4.6. Electrophysiological experiments on the hippocampus

[0600] Field recordings were performed with a concentric bipolar electrode located in the stratum radiatum of the CA1 region using an extracellular glass pipette (3.5 MΩ) filled with aCSF. Specifically, fEPSPs were evoked by Schaffer collateral (SC) / commissural fibers using a concentric bipolar electrode placed 200–300 μm from the recording pipette. Stimulation was delivered via a bipolar electrode (FHC, Bowdoin, Maine, USA) placed in the Schaffer collateral-CA1 (SC). Te SC circuits were visualized using differential interference contrast (DIC) microscopy at 4× magnification and confirmed by the ability to evoke brief, constant field excitatory postsynaptic potentials (fEPSPs) at CA1 synapses by SC input stimulation. In all fEPSP experiments, a test stimulus was measured (30 μA) before starting every experiment, and a test pulse stimulation intensity that evoked 50% of the maximum fEPSP was used. Baseline synaptic responses were recorded for 30 min, followed by induction of long-term potentiation (LTP) by high-frequency stimulation (HFS, 100 Hz, train of 4, duration of 1 s, interval between trains of 20 s). Recordings were made every 10 s for 1 h using a multiclamp 700A amplifier (Molecular Devices, San Jose, CA, USA), digitized at 10 kHz on a Digidata 1440 and filtered at 2 kHz, and pClamp 10.0 software (Molecular Devices, San Jose, CA, USA). Measurement parameters were as follows:

[0601] Measurement parameters

[0602] JPEG2025512899000047.jpg65146

[0603] Excitatory postsynaptic potentials (EPSPs) were recorded by whole-cell patch clamp technique using a patch clamp amplifier (Molecular Devices, USA). The measurement electrodes were borosilicate glass capillaries with resistances of 3.5 MΩ to 4.2 MΩ when filled with the electrode's internal solution. The brain slices in the chamber were placed on a vertical microscope, fixed with an anchor, and the recording aCSF solution was allowed to flow at a rate of 5 to 10 mL / min by gravity. The stimulator electrode was placed in the SC region, and connections in the CA1 region were located and recorded. As the stimulation intensity was increased, points were recorded until the fEPSP did not increase or a pop spike occurred (I / O curve). Baseline synaptic responses were recorded for 20 min, followed by induction of long-term potentiation (LTP) by high-frequency stimulation (HFS, 100 Hz, 4 trains, 1 s duration, 20 s intertrain interval / TBS; 100 Hz, 10 trains, 50 ms duration, 200 ms intertrain interval) followed by recording for 60 min (LTP).

[0604] Example 4.7. Data Analysis

[0605] To perform potency analysis, the Clampfit program (Axon Instruments) was used. A baseline was set by specifying an interval between 0 and 100 ms. Traces were averaged by selecting "On" currents appearing within 110 to 200 ms. The negative current amplitude (pA) of the average trace was confirmed.

[0606] GABA / AMPA ratios were analyzed using Clampfit. For AMPA currents, a baseline was set by specifying an interval between 0 and 100 ms. Traces from 110 to 200 ms were averaged. The negative current amplitude (pA) of the average trace was confirmed. For GABA currents, a baseline was set by specifying an interval between 0 and 110 ms. Traces from 110 to 300 ms were averaged. The positive current amplitude (pA) of the average trace was confirmed.

[0607] I / O curves were calculated using Clampfit. Baseline was set by specifying the interval between 0 and 6 ms. Traces (five each) recorded from the same stimulus were averaged. Fiber volley amplitude and 20-80% upslope of fEPSP were analyzed. For LTP, baseline was set by specifying the interval between 0 and 6 ms. 20-80% upslope of fEPSP were analyzed before and after LTP induction with HFS (or TBS).

[0608] Example 4.8. Statistical Analysis

[0609] All data are presented as mean ± SEM, and n indicates the number of cells used in each experiment. Typically, one slice was used from each animal; one recording was made from each slice. To compare mean values ​​from two groups, an unpaired Student's t-test was used, and in the case of three or more groups, one-way analysis of variance (ANOVA) was used. A p-value of less than 0.05 was considered statistically significant. Such statistical analyses were performed using Prism 9.0 (GraphPad Prism 9.1.2, GraphPad Software Inc., USA).

[0610] Dose-response curves were constructed using Graphpad Prism 9 according to the manufacturer's guidelines.

[0611] Example 4.9. Western Blotting

[0612] For preparation of thalamocortical (TC) slices, 9-10 week old male C57BL / 6J rodents (mice) were anesthetized with isoflurane. Subsequently, the brains were rapidly cooled by transcardial perfusion with ice-cold sucrose artificial cerebrospinal fluid (aCSF). The brains were isolated and placed in ice-cold sucrose aCSF. Paracoronal slices were prepared at a 50° angle to the midline with a 10° angle ramp. Slices were subsequently incubated in aCSF for 30 min at 35°C to recover.

[0613] Slices were subsequently incubated in aCSF for 1–4 h at room temperature (23–25°C) before being placed in the experimental recording chamber, containing standard aCSF saturated with 95% O2 / 5% CO2.

[0614] Composition of sucrose-based aCSF

[0615] JPEG2025512899000048.jpg162146

[0616] Recording Acsf composition (room temperature: RT)

[0617] JPEG2025512899000049.jpg162146

[0618] All electrophysiological experiments were performed at 27-29 °C. Rodents were perfused with blood from a sucrose-based aCSF solution (4 °C) using gravity. Rodent brains were isolated and sectioned at 400 um at a 50° angle (55° angle for mice) and attached to a vibratome disc so that the sectioned part was at the bottom. Brain slices were incubated at 35 °C incubating aCSF solution for 30 min or at RT incubating aCSF solution for 1 h. Mouse cortical tissue was prepared according to the protocol described above and subsequently homogenized and lysed for protein extraction. Protein lysates were produced from the tissues using NP40 (FNN0021, Invitrogen) protein lysis buffer containing protease and phosphate inhibitors according to the manufacturer's guidelines. Protein concentrations were determined using a BCA protein assay kit (Pierce Biotechnology) with bovine serum albumin as the standard. Equal amounts of protein (20 μg per lane) were electrophoretically separated on 8% or 12% SDS-polyacrylamide gels and then blotted onto PVDF membranes (GE Healthcare). To block nonspecific binding, the membranes were incubated with 5% nonfat dry milk in PBST (150 mM NaCl, 50 mM Tris, pH 7.4, 0.05% Tween 20) at room temperature for 1 h. Anti-HDAC4 (SAB4300413, Sigma-Aldrich), anti-BDNF (ANT-010, Alomone Las), and anti-NMDAR2B (ab65783, abcam plc) antibodies were incubated overnight at 4°C in 2% blocking solution. After incubation with the antibodies and rinsing several times with PBST, the membranes were incubated with mouse anti-rabbit IgG-HRP conjugated secondary antibody (sc-2357, Santa Cruz) for 1 h. Each blot was then re-probed with an anti-β-actin antibody (sc-47778, Santa Cruz). The housekeeping protein β-actin was detected as a normalization control. The antibodies used in the methods described herein are as follows:

[0619] JPEG2025512899000050.jpg90145

[0620] Example 5. Therapeutic efficacy in PTSD

[0621] Example 5.1. Acquisition and care of animals

[0622] Animals were obtained from DBL. Animals were obtained 7 days before the start of the study. Twenty male SD rats, approximately 7 weeks old, were used. The animals were maintained in normal environmental conditions with a 12 / 12 hour light / dark cycle in a colony room and had free access to food and water. The rats were allowed to adapt to the new housing cages for 7 days. All studies were performed using these animals. All experiments were performed in accordance with the Guide for the Care and Use of Laboratory Animals, and all efforts were made to minimize animal suffering. Body weights were measured at 3-day intervals from the 7th day onwards.

[0623] Example 5.2. Rat modeling of PTSD

[0624] The protocol involves two exposures to a foot shock without the opportunity for escape.

[0625] Context A (-1st day)

[0626] Body weight was measured 3 hours before the experiment. The temperature of the experimental room was maintained at 22°C. The animals were placed in the dark experimental room for 10 minutes to allow adaptation. Each animal was placed in the first room. After 2 minutes, the door separating the two compartments was opened, allowing the animals to freely move between the two sides. After 5 minutes, 15 foot shocks of 1 mA / 2 seconds were applied at random time intervals (90 seconds to 270 seconds). After the end of the test, the animals were left in the box for 1 minute and then moved to the colony room.

[0627] Context B (Day 0)

[0628] Body weight was measured 3 hours before the experiment. The temperature of the experiment room was maintained at 22°C. The animals were placed in a dark experiment room for 10 minutes to allow them to adapt. The animals were placed in the first compartment with sandpaper attached. At this time, the door was kept closed so that the animals could not cross into the second compartment. After 10 minutes, the animals were given three shocks of 2 mA / sec. After the test was completed, the animals were left in the box for 1 minute and then moved to the colony room.

[0629] compound injection

[0630] Compounds are diluted to 25mg / kg using DMSO:PEG400:saline (5:55:40). Compounds are intraperitoneally administered every day at 10am from day 7 to day 24 (intraperitoneal administration volume is 200μl). Control group is also intraperitoneally injected in the same manner using vehicle (DMSO:PEG400:saline (5:55:40)).

[0631] Checking the freezing

[0632] For confirmation of freezing, on the 7th day after the first footshock protocol, rats were placed in the first compartment of the apparatus with sandpaper for 5 min (without footshock) and freezing behavior was recorded using the shutavoid_v1803 program (Panlab, Spain) according to the manufacturer's guidelines, with the door kept closed as in context B. Freezing was defined as the absence of skeletal muscle movement in the rat. Animals with a freezing time of less than 240 s were excluded. Excluded animals were sacrificed using CO2.

[0633] Data analysis

[0634] A camera was placed in front of the transparent acrylic panel of the electronic box to monitor the movements of all animals. Freezing was monitored for all groups every 3 days from the 7th day. Each animal was filmed for 5 minutes. The freezing time of each animal was confirmed based on the footage. The freezing time on the 7th day was set to 100%, and the freezing time for each day of the week was then converted into a percentage.

[0635] Example 6. Therapeutic efficacy in AD

[0636] Example 6.1. Acquisition and care of animals

[0637] APP / PS1 mice, an animal model of Alzheimer's disease, were obtained from Taconic Bioscience. Animals were obtained 7 days before the start of the study. Twenty male mice, approximately 15 months old, were used. The animals were maintained in normal environmental conditions with a 12 / 12-h light / dark cycle in a colony room and had free access to food and water. The mice were allowed to adapt to the new housing cages for 7 days. All studies were performed using these animals. All experiments were performed in accordance with the Guide for the Care and Use of Laboratory Animals, and all efforts were made to minimize animal suffering. Body weights were measured at 3-day intervals from the 7th day onwards.

[0638] Example 7. Selection of BnH12003

[0639] Example 7.1. In vitro cytotoxicity

[0640] The cytotoxicity of the compounds was assessed by treating cells with BnH12003 and measuring cell viability.

[0641] Measurement of cell viability in response to BnH12003 (0, 0.1, 1, 10, 100 μM) treatment in human cell lines 293T and SH-SY5Y showed that there was no significant change in cell viability across the entire concentration range, thereby confirming the cytotoxic safety of BnH12003 (Figure 1a and Figure 1b).

[0642] Example 7.2. Analysis of the efficacy of enhancing synaptic plasticity in vitro

[0643] After administration of BnH12003, the potency and GABA / AMPA ratio in the barrel cortex were measured to confirm the efficacy of the compound.

[0644] To this end, 8-week-old C57BL / 6J mice were divided into a control group (vehicle-treated group) and an experimental group (BnH12003-treated group), each consisting of five mice each. After undergoing Alzet Os pump injection surgery, BnH12003 was directly injected into layer 4 of the barrel cortex using the Os pump at a total dose of 3 μM for 2 consecutive weeks (Figures 2a and 2b).

[0645] When BnH12003 compound was administered, it was confirmed that excitatory TC synaptic potency (TC potency) was significantly increased (Figure 3b) while maintaining the balance of excitation / inhibition, as confirmed by the measurement of GABA / AMPA ratio in mouse barrel cortex (layer 4) (Figure 3a), and it was found that BnH12003 shows efficacy in improving cognitive ability by increasing synaptic plasticity. Meanwhile, 8-week-old C57BL / 6J mice were divided into a control group (vehicle administration group) and an experimental group (BnH12003 administration group) of five mice each, and a dose of 25 mg / kg was administered intraperitoneally once a day for about 11 or 12 days. As a result, it was confirmed that administration of BnH12003 can increase TC synaptic potency in the sensory cortex while maintaining the balance of excitation / inhibition (Figure 4a and Figure 4b).

[0646] Example 7.3. Analysis of the proteolytic efficacy of HDAC4

[0647] The BnH12 series according to the present invention was synthesized as a compound targeting HDAC4, and we attempted to determine whether BnH12003 degrades HDAC4 at the protein level. Barrel cortex was isolated from the mice of Example 7.2, and HDAC4 protein levels were determined with or without BnH12003 treatment. As a result, it was confirmed that HDAC4 protein levels were reduced by 60% to 80% in mice of all experimental groups compared to mice of the control group (Figure 5). These results confirmed that BnH12003 improves cognitive function by regulating HDAC4 protein levels and increasing synaptic plasticity.

[0648] Example 7.4. Analysis of efficacy in treating post-traumatic stress disorder using animal models of post-traumatic stress disorder

[0649] Since the HDAC4 gene has been shown to be closely related to the brain's fear response, we attempted to observe the therapeutic efficacy of BnH12003 administration in an animal model of post-traumatic stress disorder (PTSD).

[0650] For this purpose, SD rodents (rats) (300 g) were adapted for 3 days, and before entering the experiment, they were placed in a chamber for 5 minutes and photographed to test the freezing of the experimental rats. In a chamber surrounded by rough sandpaper, electrical stimulation was randomly given to the soles of the feet with 15 shocks, 1 mA intensity, and 2 s duration at intervals of 90 s to 270 s. The next day, in a chamber surrounded by rough sandpaper, electrical stimulation was given to the soles of the feet every 3 s with 3 shocks every s, 2 mA intensity, and 2 s duration. By checking the freezing time on the 7th day, PTSD rodents (rats) were selected. The selected rodents (rats) were divided into a control group and an experimental group of 12 each, and 1 μM BnH12003 was administered to the ventricles of the rodents (rats) using an Os pump. On the 10th day after the start of the rodent (rat) PTSD model creation, saline was intraperitoneally administered to the control group, and BnH12003 (25mg / kg) was intraperitoneally administered to the experimental group, after which the freezing time was checked at 3-day intervals. The freezing time was checked a total of 8 times, and the reduction in freezing time was calculated as a percentage (%) based on the 7th day after the PTSD rodent (rat) model creation.

[0651] As a result, it was confirmed that the freezing time was significantly reduced in the experimental group administered with BnH12003 compared to the control group (Figure 6), and it was confirmed that when BnH12003 was administered to rats with traumatic memories (fear memories), it had the effect of reducing or eliminating the traumatic memories (fear memories). Therefore, it was confirmed that when the compound of the present invention is used, the learning effect of erasing traumatic memories (fear memories) or new memories (places that are no longer dangerous) is improved.

[0652] Example 7.5. Analysis of the efficacy of enhancing ex vivo synaptic plasticity in animal models of Alzheimer's disease

[0653] We aimed to confirm the efficacy of the compound against Alzheimer's disease by measuring the potency and GABA / AMPA ratio in the hippocampus after administration of BnH12003 to APP / PS1 mice (Taconic Bioscience), an animal model of Alzheimer's disease, and in particular to compare the effects of BnH12003 and Solanezumab administered alone and in combination. To this end, BnH12003 and Solanezumab (Creative Biolabs, USA) were administered to the hippocampus at doses of 3 μM and 200 μg, respectively, by Os pump ventricular infusion as follows: We observed the I / O curve changes of field excitatory postsynaptic potentials (EPSPs) in the CA1 region of the Schaffer collateral circuit in the hippocampus, and confirmed that in vivo administration of BnH12003 increased synaptic efficacy (Figure 7a). In addition, we measured synaptic long-term potentiation (LTP) of EPSPs in the CA1 region of the Schaffer collateral circuit in the hippocampus, and confirmed that LTP in the excitatory SC pathway was also significantly increased (Fig. 7b).

[0654] Since the increase in I / O curve and LTP indicates an increase in synaptic plasticity, these results indicate that the combination of BnH12003 and solanezumab increases the synaptic potency of the SC pathway in the hippocampus, which indicates that BnH12003 is effective in treating Alzheimer's disease. In this case, the combination of BnH12003 and solanezumab showed better effects than either BnH12003 or solanezumab administered alone.

[0655] On the other hand, we attempted to confirm the efficacy of BnH12003 against Alzheimer's disease by administering it to APP / PS1 mice and measuring the potency and GABA / AMPA ratio in the barrel cortex, and in particular, to compare the effects of BnH12003 administered alone and in combination with solanezumab.

[0656] When BnH12003 was administered to APP / PS1 mice, in vivo administration of BnH12003 increased excitatory TC synaptic potency (TC potency) while maintaining the excitation / inhibition balance (Figure 8a), as confirmed by the measurement of the GABA / AMPA ratio (Figure 8b). Since an increase in TC potency indicates increased synaptic plasticity, these results indicate that administration of the BnH12003 compound increases TC synaptic potency in the sensory cortex while maintaining the excitation / inhibition balance, which indicates that BnH12003 is effective in treating Alzheimer's disease. In this case, the co-administration of BnH12003 and solanezumab showed significantly better effects than the administration of BnH12003 and solanezumab alone.

[0657] Example 8. Surface plasmon resonance assay

[0658] The compounds of the present invention were selected for their ability to interact with HDAC4. To carry out the test, a surface plasmon resonance assay was used. Samples were prepared and loaded onto a Biacore T200 as follows:

[0659] JPEG2025512899000051.jpg63146

[0660] Samples were fixed under the following conditions: Protein stocks were diluted in 10 mM sodium acetate. 1 M ethanolamine (pH 8.5) was used as the blocking agent.

[0661] JPEG2025512899000052.jpg29146

[0662] FIG. 9 shows the binding kinetics of BnH12003 to HDAC4.

[0663] Example 9. Effect of compounds on HDAC catalytic activity

[0664] Compounds were resuspended in 100 mM DMSO and titrated to 10-volume IC in 3-fold serial dilutions starting at 100 μM against HDACs 1–9 and 11. 50 As reference compounds, Trichostatin A (TSA) and TMP269 were tested at 10 μM or 10-volume IC in 3-fold serial dilutions starting from 1 μM. 50 Tested with.

[0665] A 10 μM fluorescent peptide of p53 residues 379-382 (RHKK(Ac)AMC) was used as a substrate for HDACs 1, 2, 3, and 6. A 50 μM fluorescent HDAC class 2a substrate (trifluoroacetyl lysine) was used as a substrate for HDACs 4, 5, 7, 9, and 11. A 100 μM fluorescent peptide of p53 residues 379-382 (RHK(Ac)K(Ac)AMC) was used as a substrate for HDAC 8.

[0666] IC using the GraphPad Prism 4 program based on a sigmoidal dose-response equation 50 Values ​​were calculated and the concentration of the blank (DMSO) value was entered as 1.00E-12 M for curve fitting. Curve fitting was performed for compound data sets that showed less than 65% enzyme activity at the highest concentration of compound tested.

[0667] Experiments were performed in duplicate. BnH12003 did not show any inhibitory effect on the catalytic activity of HDACs 1-9 and 11.

[0668] Example 10. Expression of BDNF and GluN2B and effects on thalamocortical EPSC potency

[0669] When HDAC4 protein expression was inhibited by transfection with lentiviral HDAC4 shRNA, BDNF and GluN2B expression was significantly increased in SH-SY5Y cells (Figures 10a and 10b). Similarly, BnH12003 (1 μM) increased BDNF and GluN2B expression in SH-SY5Y cells after 6 and 24 h of application (Figures 11a and 11b). BnH12003 significantly inhibited HDAC4 protein expression after oral and intraperitoneal administration, respectively, in mouse barrel cortex, and such inhibitory effects were maintained for at least 10 days (Figures 12a and 12b).

[0670] Example 11. In vitro cytotoxicity of various BnH12 series compounds

[0671] The cytotoxicity of the compounds was evaluated by treating cells with various BnH12 series compounds and measuring cell viability using the experimental procedure described in Example 2. As a result, for almost all compounds, cell viability of 85% or more was observed at a concentration of 10 uM, and even when the concentration was increased to 100 uM, cell viability of 80% or more was observed.

[0672] [Table 4]

[0673] JPEG2025512899000054.jpg219139

[0674] JPEG2025512899000055.jpg126146

[0675]

[0676] The cell viability was measured using 293T cell line at various compound concentrations (10 or 100 μM), and it was shown that most of the compounds did not significantly change the cell viability within the two concentration ranges.

[0677] Example 12. Confirmation of efficacy of various BnH12 series compounds

[0678] Using the experimental procedures described in Example 3, we confirmed whether various BnH12 series compounds increased GluN2B expression, and the results are shown in Table 5. These in vitro experimental results are shown as relative values ​​to the effect of the group not treated with the compound (vehicle), which was set as 1.

[0679] [Table 5]

[0680] JPEG2025512899000057.jpg105146

[0681]

[0682] As can be seen from Table 5, the representative compounds of the present invention showed superior effects compared to the vehicle. In particular, BnH12003, BnH12046, BnH12057, BnH12067, BnH12091, BnH12092, BnH12095, BnH12097, and BnH12098 showed superior effects compared to other compounds.

Claims

1. A compound of the following chemical formula (1), a stereoisomer thereof, or a pharma- ceutically acceptable salt thereof: In the above formula, R １ is an aryl or heteroaryl, or combines with an adjacent phenyl group to form a saturated or partially saturated heterocycle, and one or more hydrogen atoms in the aryl, heteroaryl, or heterocycle are unsubstituted or substituted with a halogen atom, C １－６ Alkyl, C ３－１２ Cycloalkyl, C １－６ alkoxy, aryl, heteroaryl, amino, or nitro, the substituents being unsubstituted or substituted with one or more halogen atoms or C １－６ is substituted with alkyl, R ２ is hydrogen, C １－６ alkyl or aryl; R ３ is hydrogen, carboxyl, N-hydroxycarboxamide, or carboxamide, and the hydrogen in said group is C １－６ may be substituted with alkyl; n is 0 or 1; m is 0 to 6.

2. R １ is phenyl, pyridinyl, pyridazinyl, pyrimidinyl, pyrazinyl, furanyl, thiophenyl, pyrrolyl, imidazolyl, or oxazolyl, or is joined to an adjacent phenyl group to form quinoxalinyl or dihydrobenzodioxinyl; 2. The compound of claim 1, a stereoisomer thereof, or a pharma- ceutically acceptable salt thereof.

3. R １ Among them, one or more hydrogen atoms are halogen atoms, one or more halogen atom-substituted or unsubstituted C １－６ Alkyl or C ３－１２ Cycloalkyl, C １－６ Alkoxy, phenyl, pyridinyl, pyridazinyl, pyrimidinyl, one or more C １－６ alkyl-substituted or unsubstituted amino, or nitro-substituted; 3. The compound of claim 2, a stereoisomer thereof, or a pharma- ceutically acceptable salt thereof.

4. R １ is phenyl, biphenyl, pyridin-4-yl, pyridin-3-yl, pyridin-2-yl, methylpyridin-3-yl, 6-methoxy-pyridin-3-yl, 6-cyclopropyl-pyridin-3-yl, trifluoromethylpyridin-3-yl, phenylpyridin-3-yl, pyridazin-4-yl, pyridazin-3-yl, pyridazin-2-yl, pyrimidin-2-yl, pyrimidin-4-yl, pyrimidin-5-yl, methylpyrimidin-5-yl, 5-fluoropyrimidin-2-yl, 2-(trifluoromethyl)pyrimidin-5-yl, 2-cyclopropylpyrimidin-5-yl, pyrazin-2-yl, thiophen-2-yl, furan-2-yl, oxazol-2-yl, 3-nitrophenyl, 4-nitrophenyl, 3-aminophenyl, 4-aminophenyl, or 4-diethylaminophenyl; 2. The compound of claim 1, a stereoisomer thereof, or a pharma- ceutically acceptable salt thereof.

5. R ２ is H or C １－６ is alkyl, 2. The compound of claim 1, a stereoisomer thereof, or a pharma- ceutically acceptable salt thereof.

6. R ３ is H, carboxyl, methylcarboxyl, ethylcarboxyl, N-hydroxycarboxamide, or carboxamide; 2. The compound of claim 1, a stereoisomer thereof, or a pharma- ceutically acceptable salt thereof.

7. The stereoisomer is an optical isomer.

2. The compound of claim 1, a stereoisomer thereof, or a pharma- ceutically acceptable salt thereof.

8. The stereoisomer is the (3R,6S)-enantiomer.

2. The compound of claim 1, a stereoisomer thereof, or a pharma- ceutically acceptable salt thereof.

9. The compound is selected from the following compounds:

2. The compound of claim 1, its stereoisomer, or a pharma- ceutically acceptable salt thereof: 。

10. The compounds are BnH12003, BnH12017, BnH12045, BnH12046, BnH12047, BnH12048, BnH12051, BnH12053, BnH12057, BnH12058, BnH12064, BnH12066, BnH12067, BnH12071 , BnH12072, BnH12078, BnH12085, BnH12086, BnH12088, BnH12089, BnH12090, BnH12091, BnH12092, BnH12093, BnH12095, BnH12097, and BnH12098; 10. The compound of claim 9, a stereoisomer thereof, or a pharma- ceutically acceptable salt thereof.

11. The compound is an inhibitor of HDAC.

2. The compound of claim 1, a stereoisomer thereof, or a pharma- ceutically acceptable salt thereof.

12. The HDAC is HDAC4.

12. The compound of claim 11, a stereoisomer thereof, or a pharma- ceutically acceptable salt thereof.

13. The compound increases synaptic plasticity.

2. The compound of claim 1, a stereoisomer thereof, or a pharma- ceutically acceptable salt thereof.

14. A pharmaceutical composition comprising a compound according to any one of claims 1 to 13, a stereoisomer thereof, or a pharma- ceutically acceptable salt thereof as an active ingredient. Pharmaceutical compositions.

15. The composition is for the prevention or treatment of a neurological or neurodegenerative disorder. The pharmaceutical composition of claim 14.

16. The compound, its stereoisomer, or its pharma- ceutically acceptable salt is delivered to the brain at a concentration of about 0.1 to about 3 μM. The pharmaceutical composition of claim 14.

17. The disorder is Alzheimer's disease. The pharmaceutical composition of claim 15.

18. The disorder is PTSD. The pharmaceutical composition of claim 15.

19. further comprising a second therapeutic agent, The pharmaceutical composition of claim 14.

20. The second therapeutic agent is solanezumab.

20. The pharmaceutical composition of claim 19.

21. A method for the prevention or treatment of a neurological or neurodegenerative disorder, comprising administering to an individual in need thereof a compound according to any one of claims 1 to 13, a stereoisomer thereof, or a pharma- ceutically acceptable salt thereof.

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