Modulators of PrP and uses thereof

Abstract

The present invention relates to compounds capable of modulating the activity of normal prion protein (PrP) and their use for the treatment of immune and neurodegenerative diseases.

Description

[Technical Field]

[0001] FIELD OF THE INVENTION The present invention relates to compounds capable of modulating the activity of cellular prion protein (PrP) and their use for the treatment of neurodegenerative and immune diseases. In particular, the compounds of the present invention are useful for the treatment of Alzheimer's disease, prion diseases, multiple sclerosis, autoimmune encephalitis, Parkinson's disease, inflammatory bowel disease, and Crohn's disease. [Background technology]

[0002] Background of the Invention Aging is associated with a wide range of molecular, cellular, and functional changes, particularly affecting the integrity of the nervous system. One of the major processes altered by aging is protein folding. When proteins misfold, they adopt alternative conformations that can seed a cascade of molecular events, ultimately leading to neuronal dysfunction and death. Indeed, a wide range of age-related disorders are associated with protein misfolding and aggregation in the brain. Examples include common disorders such as Parkinson's disease and Alzheimer's disease, as well as rarer disorders such as prion diseases. 1(Non-Patent Document 1). Alzheimer's disease is the most common dementia in the elderly population, currently affecting approximately 36 million people worldwide. As the population ages, the number of cases will increase dramatically in the coming decades, resulting in devastating medical and socioeconomic impacts. According to the amyloid cascade hypothesis, Alzheimer's disease results from the accumulation in the brain of Aβ peptides, which consist of 40 to 42 amino acids and are cleaved products of the amyloid precursor protein (APP). The majority of Alzheimer's disease cases manifest as late-onset, sporadic forms. However, approximately 5% of cases are inherited in an autosomal dominant manner. These forms, collectively known as familial Alzheimer's disease, are associated with at least 230 mutations in the genes encoding APP or presenilin (PS1 or PS2). 2 (Non-Patent Document 2). This mutation is thought to promote the accumulation of Aβ peptides in the brain by increasing their production and decreasing their clearance (excretion). Aβ peptides spontaneously form various polymers ranging from small soluble oligomers to large insoluble fibrils. A large body of evidence suggests that the primary cause of synaptic dysfunction underlying cognitive decline in Alzheimer's disease is soluble Aβ oligomers, rather than fibrillar aggregates. 3 (Non-Patent Document 3). Aβ oligomers are thought to act by binding to cell surface receptors that transmit harmful effects to synapses. Identification of such receptor sites has therapeutic significance, as they are potential targets for pharmacological intervention. In recent years, a new candidate receptor for Aβ oligomers, the normal (cellular) prion protein (PrPC), has emerged. 4 (Non-Patent Document 4) PrP is an endogenous cell surface glycoprotein with unknown function that plays a central role in transmissible neurodegenerative disorders commonly referred to as prion diseases.

[0003] PrPC was initially discovered as playing a central role in transmissible spongiform encephalopathies (also known as prion diseases) and has been postulated to be involved in several other neuropathologies, including Alzheimer's disease and Parkinson's disease, by acting as a toxicity-transducing receptor for various misfolded protein isoforms. Interestingly, PrPC has also been reported to exert important functions outside the nervous system, particularly in the immune system, where this protein has emerged as a key factor in myelin homeostasis. Consistent with these concepts, further studies have revealed that the absence of PrPC exacerbates inflammatory damage in various laboratory models of cerebral ischemia, brain trauma, experimental autoimmune encephalomyelitis (EAE), and experimental colitis.

[0004] Prion diseases can be sporadic, inherited, or acquired and are caused by the conformational transformation of PrP into a misfolded isoform (abnormal (scrapie) PrP or PrPSc) that accumulates in the central nervous system of affected individuals. PrPSc is an infectious protein (prion) that self-propagates by binding to PrPc, causing conformational rearrangement into new PrPSc molecules. 5 (Non-Patent Document 5). A large body of evidence has demonstrated the distinction between prion infectivity and toxicity, suggesting that the physiological function of PrP may be altered upon binding to PrP, potentially delivering neurotoxic signals. Indeed, genetic depletion of neuronal PrP in prion-infected mice has been shown to reverse neuronal loss and clinical progression, despite the continued production of PrP by surrounding astrocytes. 6(Non-Patent Document 6). Thus, the presence of PrP on the neuronal cell surface is important not only for supporting the propagation of PrP but also for transducing its neurotoxicity. 7,8 (Non-Patent Documents 7, 8). This conclusion has recently received unexpected support from data on Aβ oligomers. PrPc emerged from an expression cloning screen as a receptor capable of binding Aβ oligomers with nanomolar affinity. Importantly, PrPc was also found to be a mediator of Aβ-induced synaptotoxicity. 4 (Non-Patent Document 4). Supporting this conclusion, hippocampal slices from PrP knockout (KO) mice were shown to be resistant to Aβ oligomer-induced long-term potentiation (LTP) suppression, an in vitro correlate of memory and synaptic function. Consistent with this, application of anti-PrP antibodies was shown to prevent Aβ-induced synaptic dysfunction in hippocampal slices. 9 (Non-Patent Document 9). Finally, PrP was required for both the cognitive impairment and reduced survival observed in a transgenic mouse model of Alzheimer's disease. 10 (Non-Patent Document 10). Subsequent studies have extended this finding by finding that some Aβ assemblies, including neurotoxic Aβ oligomers, bind with high affinity to PrP via two sites in the unstructured N-terminal tail of the protein (residues 23-27 and 95-105). 11 (Non-Patent Document 11). This interaction unleashes toxic signaling, including activation of metabotropic glutamate receptor 5 (mGluR5), tyrosine kinase Fyn, and phosphorylation of the NR2B subunit of the NMDA receptor, ultimately resulting in dysregulation of receptor function, excitoxicity, and dendritic spine retraction. 12 (Non-Patent Document 12). Other recent studies have provided evidence that PrP can mediate not only the toxicity of Aβ oligomers but also the toxicity of other β-sheet-rich protein conformers, including α-synuclein, which is involved in Parkinson's disease. 13-15(Non-Patent Documents 13-15). These results suggest that misfolded assemblies of several different pathogenic proteins may exert their effects by interfering with, enhancing, or altering the normal activity of PrP. 8 (Non-Patent Document 8). This conclusion highlights the close relationship between the role of PrPC in several neurodegenerative diseases and its physiological functions. Several activities have been attributed to PrPC in the nervous system, mostly based on subtle abnormalities detected in mice or cells depleted of PrPC. These include roles in neuroprotection, synaptic homeostasis, neuronal excitability, and memory formation. 16 (Non-patent Document 16). In recent years, it has been shown that PrP also plays important functions outside the nervous system, particularly in the immune system. 17 (Non-Patent Document 17). PrPC appears to be protective in autoimmune colitis. Inflammatory bowel disease is induced by dextran sulfate sodium (DSS) and is more severe in PrP0 / 0 mice than in wild-type mice. Thus, overexpression of PrPC significantly attenuates DSS-induced colitis. Upon MHC / peptide-driven interactions between T cells and dendritic cells (DCs), PrPC migrates to the immune synapse and exerts distinct effects on T cell proliferation and cytokine production, as revealed by ablation or antibody masking on the DC or lymphocyte side of the synapse. 18 DCs are professional antigen-presenting cells (APCs) and highly plastic cells that play an important role in the differentiation of helper T (Th) cells, and are therefore involved in the induction of both autoimmunity and tolerance. 19(Non-Patent Document 19). Surprisingly, the present authors found that selected DC subsets express high levels of PrP. Furthermore, recent data have revealed that EAE is exacerbated in PrP-deficient mice, suggesting that this protein may act as a regulatory molecule and that PrP-deficient cells may become more inflammatory and aggressive to the central nervous system. Based on these results, the present inventors hypothesized that pharmacological targeting of PrP may activate protective immunoregulatory effects in multiple sclerosis (MS).

[0005] A highly robust and rapid assay for detecting the spontaneous toxicity of mutant PrP in cell culture, termed the cell-based drug assay (DBCA), has been previously described. 20 (Non-Patent Document 20). This novel assay was recently used to identify small molecules that can eliminate mutant PrP activity. 21,22 (Non-Patent Documents 21, 22). Importantly, one derivative of such a molecule (designated SM), resulting from several cycles of chemical optimization, rescued Aβ-induced synaptic dysfunction in primary hippocampal neurons and rescued prion-induced electrophysiological abnormalities in mouse brain slices. Taking these data together, the inventors hypothesized that pharmacological modulation of PrP activity might have therapeutic benefits in multiple sclerosis (MS), a neurodegenerative disorder characterized by progressive myelin loss. Furthermore, in a mouse model of EAE, systemic administration of such PrP modulators was found to significantly reduce disease severity compared with untreated controls. Within this conceptual framework, there remains a need for compounds that can modulate the activity of PrP. [Prior art documents] [Non-patent literature]

[0006] [Non-Patent Document 1] Chiti, F. & Dobson, CM Protein Misfolding, Amyloid Formation, and Human Disease: A Summary of Progress Over the Last Decade. Annu Rev Biochem 86, 27-68, doi:10.1146 / annurev-biochem-061516-045115 (2017). [Non-patent document 2] Selkoe, DJ & Hardy, J. The amyloid hypothesis of Alzheimer's disease at 25 years. EMBO Mol Med 8, 595-608, doi:10.15252 / emmm.201606210 (2016). [Non-patent document 3] Walsh, DM et al. Naturally secreted oligomers of amyloid beta protein potently inhibit hippocampal long-term potentiation in vivo. Nature 416, 535-539, doi:10.1038 / 416535a (2002). [Non-patent document 4] Lauren, J., Gimbel, DA, Nygaard, HB, Gilbert, JW & Strittmatter, SM Cellular prion protein mediates impairment of synaptic plasticity by amyloid-beta oligomers. Nature 457, 1128-1132, doi:10.1038 / nature07761 (2009). [Non-Patent Document 5] Prusiner, SB Prions. Proc Natl Acad Sci USA 95, 13363-13383, doi:10.1073 / pnas.95.23.13363 (1998).

Non-licensed Document 6

Non-licensed Document 7

Non-licensed Document 8

Non-licensed literature 9

Non-licensed literature 10

Non-licensed Document 11

Non-licensed Document 12

Non-licensed Document 13

Non-licensed Document 14

Non-licensed Document 15

Non-licensed Document 16

Non-licensed Document 17

Non-licensed Document 18

Outdoor Tools 19

Outdoor Tools20

Direct Environment21

Optional Website22

[0007] Summary of the Invention In the present invention, we have surprisingly found that appropriately functionalized thiazine-dioxide scaffolds provide a series of compounds capable of abolishing mutant PrP activity. The results obtained within this invention clearly demonstrate that the compounds may represent new therapeutic options for several different pathologies, such as prion diseases, Alzheimer's disease, autoimmune encephalitis, and MS.

[0008] The object of the present invention is to provide a compound of the following general formula (I), and any stereoisomers, pharmaceutically acceptable salts, solvates and hydrates thereof, for use in the treatment of neurodegenerative or immune diseases, preferably for use in the treatment of Alzheimer's disease, prion diseases, multiple sclerosis, autoimmune encephalitis, Parkinson's disease, inflammatory bowel disease and Crohn's disease: The aim is to provide :

[0009] [ka]

[0010] [In the formula, A is a benzene ring or a 5- or 6-membered heteroaromatic ring; B has the following general structure:

[0011] [ka]

[0012] is a benzene ring, Or, B is a 5- or 6-membered heteroaromatic ring optionally substituted by one or more substituents, each of which is independently hydrogen, halogen, nitro, cyano, thiol, C 1-4 Alkyl, haloalkyl, O-haloalkyl, OC 1-4 Alkyl, NHC 1-4 Alkyl, C(=O)C 1-6 Alkyl, C(=O)OC 1-6 Alkyl, C(=O)NHC 1-4 Alkyl, hydroxy, SC 1-4 Alkyl, OC 1-4 alkylamino; W is C(=O), C(=S), CH2 or absent; Y is selected from CH2, SO2, SO, S, C(=O), PO2 and NR4, preferably Y is selected from CH2, SO2, SO, C(=O) and NR4; Z is N or CH; X1 and X2 are each independently hydrogen, halogen, nitro, cyano, thiol, C 1-4 Alkyl, haloalkyl, O-haloalkyl, OC 1-4 Alkyl, NHC 1-4 Alkyl, C(=O)C 1-6 Alkyl, C(=O)OC 1-6 Alkyl, C(=O)NHC 1-4 Alkyl, hydroxy, SC 1-4 Alkyl, OC 1-4 selected from alkylamino, OH, pyrrolidine, piperidine, morpholine, piperazine, N-methylpiperazine; X3 is hydrogen, methyl, ethyl, isopropyl or benzyl; X4 and X5 are independently hydrogen, C 1-3 selected from alkyl, haloalkyl, halogen, cycloalkyl, amino, hydroxy, cyano, and nitro; and n is 0, 1, 2, 3, or 4; Or, residues X3 and X4 together form a single bond or C 1-4represents an alkanediyl, and X3 and X4 form a 5- or 6-membered heterocycle together with the bridging atoms to which they are bonded; R1, R2, R 2a and R3 are each independently hydrogen, halogen, nitro, cyano, hydroxy, thiol, C 1-4 Alkyl, haloalkyl, O-haloalkyl, OC 1-4 Alkyl, NHC 1-4 Alkyl, C(=O)C 1-6 Alkyl, C(=O)OC 1-6 Alkyl, C(=O)NHC 1-4 Alkyl, OC 1-4 Alkylamino, SC 1-4 alkyl, R4 is hydrogen, C 1-4 Alkyl, C 1-4 Aminoalkyl, C 1-4 Hydroxyalkyl, C 1-4 Nitroalkyl, C 1-4 Thioalkyl, C 1-6 haloalkyl; Q is C 1-8 Alkyl, C 1-8 selected from alkenyl, cycloalkyl, heterocycloalkyl, aryl ring, heteroaromatic ring, - Said C 1-8 Alkyl is hydroxy, OC 1-4 Alkyl, NHC 1-4 Alkyl, N(C 1-4 alkyl)2, NH(C=O)C 1-4 optionally substituted with alkyl, aryl, heteroaryl, heterocycloalkyl, cycloalkyl, or cycloalkenyl, each of which is optionally substituted with methyl, halogen, or hydroxy; - the cycloalkyl and the heterocycloalkyl are each selected from OH, OSO2R5, C 1-3 alkyl, NR6R7, where R5 is selected from hydrogen, phenyl, heteroaryl, aminophenyl and nitrophenyl, and wherein R6 and R7 are each independently selected from H, methyl, C(=O)CH3, and SO2CH3; - the aryl ring or the heteroaromatic ring is each substituted with one or more substituents; The substituents may be halogen, nitro, cyano, thiol, C 1-4 Alkyl, haloalkyl, O-haloalkyl, OC 1-4 Alkyl, NH2, NHSO2C 1-4 Alkyl, NHC 1-4 Alkyl, C(=O)C 1-6 Alkyl, C(=O)OC 1-6 Alkyl, C(=O)NHC 1-4 Alkyl, hydroxy, SC 1-4 Alkyl, OC 1-4 alkylamino. (However, the following compounds

[0013] [ka] is not included.)

[0014] A further object of the present invention is to provide a compound of formula (I) and any stereoisomers, pharmaceutically acceptable salts, hydrates and solvates thereof for use in the treatment of multiple sclerosis, autoimmune encephalitis or immune diseases. to provide and preferably the immune disease is selected from inflammatory bowel disease and Crohn's disease:

[0015] [ka]

[0016] [In the formula, A is a benzene ring or a 5- or 6-membered heteroaromatic ring; B has the following general structure:

[0017] [ka]

[0018] is a benzene ring, Or, B is a 5- or 6-membered heteroaromatic ring optionally substituted by one or more substituents, each of which is independently hydrogen, halogen, nitro, cyano, thiol, C 1-4 Alkyl, haloalkyl, O-haloalkyl, OC 1-4 Alkyl, NHC 1-4 Alkyl, C(=O)C 1-6 Alkyl, C(=O)OC 1-6 Alkyl, C(=O)NHC 1-4 Alkyl, hydroxy, SC 1-4 Alkyl, OC 1-4 alkylamino; W is C(=O), C(=S), CH2 or absent; Y is selected from CH2, SO2, SO, S, C(=O), PO2 and NR4, preferably Y is selected from CH2, SO2, SO, C(=O) and NR4; Z is N or CH; X1 and X2 are each independently hydrogen, halogen, nitro, cyano, thiol, C 1-4 Alkyl, haloalkyl, O-haloalkyl, OC 1-4 Alkyl, NHC 1-4 Alkyl, C(=O)C 1-6 Alkyl, C(=O)OC 1-6 Alkyl, C(=O)NHC 1-4 Alkyl, hydroxy, SC 1-4 Alkyl, OC 1-4 selected from alkylamino, OH, pyrrolidine, piperidine, morpholine, piperazine, N-methylpiperazine; X3 is hydrogen, methyl, ethyl, isopropyl or benzyl; X4 and X5 are independently hydrogen, C 1-3selected from alkyl, haloalkyl, halogen, cycloalkyl, amino, hydroxy, cyano, and nitro; and n is 0, 1, 2, 3, or 4; or residues X3 and X4 together form a single bond or C 1-4 represents alkanediyl, and X3 and X4 together with the bridging atoms to which they are bonded form a 5- or 6-membered heterocycle; R1, R2, R 2a and R3 are each independently hydrogen, halogen, nitro, cyano, hydroxy, thiol, C 1-4 Alkyl, haloalkyl, O-haloalkyl, OC 1-4 Alkyl, NHC 1-4 Alkyl, C(=O)C 1-6 Alkyl, C(=O)OC 1-6 Alkyl, C(=O)NHC 1-4 Alkyl, OC 1-4 Alkylamino, SC 1-4 alkyl, R4 is hydrogen, C 1-4 Alkyl, C 1-4 Aminoalkyl, C 1-4 Hydroxyalkyl, C 1-4 Nitroalkyl, C 1-4 Thioalkyl, C 1-6 haloalkyl; Q is C 1-8 Alkyl, C 1-8 selected from alkenyl, cycloalkyl, heterocycloalkyl, aryl ring, heteroaromatic ring, - Said C 1-8 Alkyl is hydroxy, OC 1-4 Alkyl, NHC 1-4 Alkyl, N(C 1-4 alkyl)2, NH(C=O)C 1-4 optionally substituted with alkyl, aryl, heteroaryl, heterocycloalkyl, cycloalkyl, or cycloalkenyl, each of which is optionally substituted with methyl, halogen, or hydroxy; - the cycloalkyl and the heterocycloalkyl are each selected from OH, OSO2R5, C 1-3 alkyl, NR6R7, where R5 is selected from hydrogen, phenyl, heteroaryl, aminophenyl and nitrophenyl, and wherein R6 and R7 are each independently selected from H, methyl, C(=O)CH3, and SO2CH3; - the aryl ring or the heteroaromatic ring is each substituted with one or more substituents; The substituents may be halogen, nitro, cyano, thiol, C 1-4 Alkyl, haloalkyl, O-haloalkyl, OC 1-4 Alkyl, NH2, NHSO2C 1-4 Alkyl, NHC 1-4 Alkyl, C(=O)C 1-6 Alkyl, C(=O)OC 1-6 Alkyl, C(=O)NHC 1-4 Alkyl, hydroxy, SC 1-4 Alkyl, OC 1-4 alkylamino.

[0019] Preferably, in the compound of formula (I), A is benzene, and / or Y is SO, and / or W is C(=O) or CH, and / or Z is N, and / or X and X are H.

[0020] Even more preferably, the compounds for use according to the invention have the following general formula (II):

[0021] [ka]

[0022] (In the formula, X1, X2, X3, R1, R2, R 2a , R3 and Q are as defined above.

[0023] In one preferred embodiment, the compound for use in the present invention is selected from the following list:

[0024] [ka]

[0025] [ka]

[0026] [ka]

[0027] [ka]

[0028] A further object of the present invention is to provide a compound of the following general formula (III), and any stereoisomers, pharmaceutically acceptable salts, hydrates and solvates thereof: The aim is to provide :

[0029] [ka]

[0030] [In the formula, A is a benzene ring or a 5- or 6-membered heteroaromatic ring; B is a benzene ring of the general structure:

[0031] [ka]

[0032] (In the formula, R1, R2 and R3 are each independently hydrogen, halogen, nitro, cyano, thiol, C 1-4 Alkyl, haloalkyl, O-haloalkyl, OC 1-4 Alkyl, NHC 1-4Alkyl, C(=O)C 1-6 Alkyl, C(=O)OC 1-6 Alkyl, C(=O)NHC 1-6 Alkyl, OC 1-4 Alkylamino, Hydroxy, SC 1-4 alkyl, R 2a are hydrogen, CF3, F, OH, OC 1-4 Alkyl, SC 1-4 Alkyl, OC 1-4 alkylamino, however, -R 2a is hydrogen or F, then R2 and / or R3 are each independently selected from F, Cl, Br, CF3, OMe, OH; or - When R1 is halogen, R 2a is hydrogen, and R2 and R3 are each independently hydrogen, halogen, nitro, cyano, thiol, C 1-4 Alkyl, haloalkyl, O-haloalkyl, OC 1-4 Alkyl, NHC 1-4 Alkyl, C(=O)C 1-6 Alkyl, C(=O)OC 1-6 Alkyl, C(=O)NHC 1-6 Alkyl, OC 1-4 Alkylamino, Hydroxy, SC 1-4 alkyl. Or Or, B is a 5- or 6-membered heteroaromatic ring optionally substituted by one or more substituents, each of which is independently hydrogen, halogen, nitro, cyano, thiol, C 1-4 Alkyl, haloalkyl, O-haloalkyl, OC 1-4 Alkyl, NHC 1-4 Alkyl, C(=O)C 1-4 Alkyl, C(=O)OC 1-6 Alkyl, C(=O)NHC 1-4 Alkyl, hydroxy, SC 1-4 Alkyl, OC 1-4 alkylamino; W is C(=O), Y is selected from CH2, SO2, SO, S, C(=O), PO2 and NR4, preferably Y is selected from CH2, SO2, SO, C(=O) and NR4; Z is N or CH; X1 and X2 are each independently hydrogen, halogen, nitro, cyano, thiol, C 1-4 Alkyl, haloalkyl, O-haloalkyl, OC 1-4 Alkyl, NHC 1-4 Alkyl, C(=O)C 1-6 Alkyl, C(=O)OC 1-6 Alkyl, C(=O)NHC 1-4 Alkyl, hydroxy, SC 1-4 Alkyl, OC 1-4 selected from alkylamino, OH, pyrrolidine, piperidine, morpholine, piperazine, N-methylpiperazine; X3 is hydrogen, methyl, ethyl, isopropyl or benzyl; X4 and X5 are independently hydrogen, C 1-3 selected from alkyl, haloalkyl, halogen, cycloalkyl, amino, hydroxy, cyano, and nitro; and n is 0, 1, 2, 3, or 4; or residues X3 and X4 together form a single bond or C 1-4 represents alkanediyl, and X3 and X4 together with the bridging atoms to which they are bonded form a 5- or 6-membered heterocycle; R4 is hydrogen, C 1-4 Alkyl, C 1-4 Aminoalkyl, C 1-4 Hydroxyalkyl, C 1-4 Nitroalkyl, C 1-4 Thioalkyl, C 1-6 haloalkyl; Q is C 1-8 Alkyl, C 1-8 selected from alkenyl, cycloalkyl, heterocycloalkyl, aryl ring, heteroaromatic ring, - Said C 1-8 Alkyl is hydroxy, OC 1-4 Alkyl, NHC 1-4Alkyl, N(C 1-4 alkyl)2, NH(C=O)C 1-4 optionally substituted with alkyl, aryl, heteroaryl, heterocycloalkyl, cycloalkyl, or cycloalkenyl, each of which is optionally substituted with methyl, halogen, or hydroxy; - the cycloalkyl and the heterocycloalkyl are each selected from OH, OSO2R5, C 1-3 alkyl, NR6R7, where R5 is selected from hydrogen, phenyl, heteroaryl, aminophenyl and nitrophenyl, and R6 and R7 are each independently selected from H, methyl, C(=O)CH3, and SO2CH3; - the aryl ring or the heteroaromatic ring is each substituted with one or more substituents; The substituents may be halogen, nitro, cyano, thiol, C 1-4 Alkyl, haloalkyl, O-haloalkyl, OC 1-4 Alkyl, NH2, NHSO2C 1-4 Alkyl, NHC 1-4 Alkyl, C(=O)C 1-6 Alkyl, C(=O)OC 1-6 Alkyl, C(=O)NHC 1-4 Alkyl, hydroxy, SC 1-4 Alkyl, OC 1-4 alkylamino.

[0033] Preferably, in the compounds defined above, A is benzene, and / or Y is SO2, and / or Z is N, and / or X4 and X5 are H.

[0034] In one preferred embodiment, the compound of formula (III) is a compound of formula (IIIA):

[0035] [ka]

[0036] (In the formula, X1, X2, R1, R2, R 2a , R3 and Q are as defined above for general formula (III).

[0037] Even more preferably, the compound of formula (III) is selected from:

[0038] [ka]

[0039] [ka]

[0040] In one preferred embodiment of formula (I) or (II), B is selected from:

[0041] [ka]

[0042] In one embodiment of Formula (I), ring B is:

[0043] [ka]

[0044] In one preferred embodiment of formula (III), B is selected from:

[0045] [ka]

[0046] In a preferred embodiment, the compound as defined above is for medical use, preferably for the treatment of a neurodegenerative or immune disease, even more preferably for the treatment of prion diseases, Alzheimer's disease, multiple sclerosis, autoimmune encephalitis, Parkinson's disease, inflammatory bowel disease, Crohn's disease.

[0047] Another aspect of the present invention relates to a method of treating diseases that would benefit from modulation of PrP activity, such as prion diseases, Alzheimer's disease, multiple sclerosis, autoimmune encephalitis, Parkinson's disease, inflammatory bowel disease, and Crohn's disease, comprising administering to a patient in need thereof a therapeutically effective amount of a compound of Formula (I), (II), or (III), wherein the limitations and definitions set forth above include any pharmaceutically acceptable salts, solvates, or stereoisomers thereof, as defined above.

[0048] A further object of the present invention is a pharmaceutical composition for use in the treatment of neurodegenerative diseases, preferably for use in the treatment of Alzheimer's disease, prion diseases, multiple sclerosis and autoimmune encephalitis, even more preferably for use in the treatment of multiple sclerosis and autoimmune encephalitis, comprising at least one compound as defined above, alone or in combination with at least one further active compound, together with at least one pharmaceutically acceptable excipient. The aim is to provide .

[0049] A further object of the present invention is a method for the synthesis of compounds of general formula (III) in which A is benzene and Y is SO2. to provide The method comprises the steps of: (a) reacting a compound of formula 1a with an aromatic or heteroaromatic amine of formula 1b in the presence of a solvent such as dichloromethane and an amine such as pyridine, trimethylamine, diethylisopropylamine, to obtain a compound of formula 2a:

[0050] [ka]

[0051] (b) reducing the nitro group of the compound of formula 2a to an amino group by hydrogenation under appropriate conditions in the presence of a Raney-Nichel catalyst or using SnCl2·2H2O to obtain a compound of formula 3a:

[0052] [ka]

[0053] (c) converting compound 3a to compound of formula 5a by a first step involving reaction with NaNO, NaOH and HCl under appropriate conditions, and a second step using Cu powder and DMSO as solvent at room temperature:

[0054] [ka]

[0055] (d) converting the compound of formula 5a to a compound of formula (I), wherein the reaction comprises at least one of the following steps: - 5a and the formula: hal-(CH2) n -C(=O)OEt or an alkylating agent of formula: hal-(CH n -Q with an alkylating agent, wherein hal is bromine or chlorine; - treatment with an amine of formula Q-NHX3 under microwave irradiation and neat conditions, - Coupling with amines of formula Q-NHX3 in the presence of a condensing agent such as TBTU and DIPEA in CH2Cl2 or using SOCl2 as a chlorinating agent.

[0056] That is, the present invention includes the following aspects.

[0057] Aspect [1]: A compound of the following general formula (I), and any stereoisomer, pharmaceutically acceptable salt, hydrate, or solvate thereof, for use in the treatment of a neurodegenerative disease or an immune disease.

[0058] [ka] [In the formula, A is a benzene ring or a 5- or 6-membered heteroaromatic ring; B has the following general structure:

[0059]

change

[0060]

change

[0061] Aspect [2]: The compound according to aspect [1], wherein the neurodegenerative disease or immune disease is Alzheimer's disease, prion disease, multiple sclerosis, autoimmune encephalitis, Parkinson's disease, inflammatory bowel disease, or Crohn's disease, and any stereoisomer, pharmaceutically acceptable salt, hydrate, or solvate thereof.

[0062] Aspect [3]: A compound of the following general formula (I), and any stereoisomer, pharmaceutically acceptable salt, hydrate, or solvate thereof, for use in the treatment of multiple sclerosis, autoimmune encephalitis, or immune diseases.

[0063]

change

change

[0064] Aspect [4]: ​​The compound according to aspect [3], wherein the immune disease is inflammatory bowel disease or Crohn's disease, and any stereoisomer, pharmaceutically acceptable salt, hydrate, or solvate thereof.

[0065] Aspect [5]: In the general formula (I), (5-1) A is benzene, and / or (5-2) Y is SO 2 and / or (5-3) W is C(=O) or CH 2 and / or (5-4) Z is N, and / or (5-5)X 4 and X 5 is H, The compound according to any one of the above aspects [1] to [4], and any stereoisomer, pharmaceutically acceptable salt, hydrate, or solvate thereof.

[0066] Aspect [6]: The following general formula (II):

[0067]

change

[0068] Aspect [7]: The following formula

[0069]

change

[0070]

change

[0071]

change

[0072] Aspect [8]: The following formula

[0073]

change

[0074] Aspect [9]: The following formula

[0075]

change

[0076] Aspect

[10] : A compound represented by the following general formula (III), and any stereoisomer, pharmaceutically acceptable salt, hydrate, or solvate thereof:

[0077]

change

[0078]

change

[0079] Aspect

[11] : In the general formula (III), (11-1) A is benzene, and / or (11-2) Y is SO 2 and / or (11-3) Z is N, and / or (11-4)X 4 and X 5 is H, The compound according to the above aspect

[10] , and any stereoisomer, pharmaceutically acceptable salt, hydrate, or solvate thereof.

[0080] Aspect

[12] : The compound represented by the general formula (III) is represented by the following formula:

[0081]

change

[0082]

change

[10] or

[11] , and any stereoisomer, pharmaceutically acceptable salt, hydrate, or solvate thereof, .

[0083] Aspect

[13] : The compound according to any one of Aspects

[10] to

[12] , wherein the compound represented by the general formula (III) is for medical use, and any stereoisomer, pharmaceutically acceptable salt, hydrate, or solvate thereof.

[0084] Aspect

[14] : The compound according to aspect

[13] , and any stereoisomer, pharmaceutically acceptable salt, hydrate, or solvate thereof, for use in treating a neurodegenerative disease or an immune disease. .

[0085] Aspect

[15] : The compound according to aspect

[14] , wherein the neurodegenerative disease or immune disease is prion disease, Alzheimer's disease, multiple sclerosis, autoimmune encephalitis, Parkinson's disease, inflammatory bowel disease, or Crohn's disease, and any stereoisomer, pharmaceutically acceptable salt, hydrate, or solvate thereof. .

[0086] Aspect

[16] : A compound for use according to any one of Aspects [1] to [9], which modulates the activity of normal prion protein (PrPC), and any stereoisomer, pharmaceutically acceptable salt, hydrate, or solvate thereof; or a compound according to any one of Aspects

[10] to

[15] , and any stereoisomer, pharmaceutically acceptable salt, hydrate, or solvate thereof.

[0087] Aspect

[17] : A compound for use according to any one of Aspects [1] to [9], and any stereoisomer, pharmaceutically acceptable salt, hydrate, or solvate thereof, administered orally or parenterally in a single dose or in divided doses in a dosage range of 0.001 to 1000 mg per kg of mammalian body weight per day, or a compound according to any one of Aspects

[10] to

[15] , and any stereoisomer, pharmaceutically acceptable salt, hydrate, or solvate thereof. .

[0088] Aspect

[18] : A pharmaceutical composition comprising at least one compound according to any one of Aspects [1] to [9], and any stereoisomer, pharmaceutically acceptable salt, hydrate, or solvate thereof, alone or in combination with at least one additional active compound, together with at least one pharmaceutically acceptable excipient, for use according to any one of Aspects [1] to [9].

[0089] Aspect

[19] : A pharmaceutical composition for use according to Aspect

[18] , which is orally administered in the form of a tablet or capsule containing 1.0 to 500 milligrams of the active ingredient.

[0090] Aspect

[20] : A pharmaceutical composition comprising at least one compound according to any one of Aspects

[10] to

[15] , and any of its stereoisomers, pharmaceutically acceptable salts, hydrates, and solvates, alone or in combination with at least one additional active compound, together with at least one pharmaceutically acceptable excipient.

[0091] Aspect

[21] : The pharmaceutical composition according to Aspect

[20] , which is a composition for use in the treatment of neurodegenerative diseases or immune diseases.

[0092] Aspect

[22] : The pharmaceutical composition according to Aspect

[21] , wherein the neurodegenerative disease or immune disease is a prion disease, Alzheimer's disease, multiple sclerosis, autoimmune encephalitis, Parkinson's disease, inflammatory bowel disease or Crohn's disease.

[0093] Aspect

[23] : A method for synthesizing a compound represented by the general formula (III) according to any one of Aspects

[10] to

[15] (wherein A is benzene and Y is SO 2 ), which method comprises the following steps: (a) Reacting a compound of formula 1a with an aromatic or heteroaromatic amine of formula 1b in the presence of a solvent such as dichloromethane and an amine such as pyridine, trimethylamine, diethylisopropylamine, etc. to obtain a compound of formula 2a:

[0094]

change

[0095]

change

[0096]

change

[0097] As used herein, "alkyl" is intended to include both branched and straight-chain saturated aliphatic hydrocarbon groups having the specified number of carbon atoms. For example, "C 1-6 "Alkyl" is defined to include groups having 1, 2, 3, 4, 5, or 6 carbons in a linear or branched arrangement, and specifically includes methyl, ethyl, n-propyl, i-propyl, n-butyl, t-butyl, i-butyl, pentyl, hexyl, and the like. Preferably, "C 1-6 Alkyl" is "C 1-4 Alkyl" or "C 1-3 It refers to "C alkyl". 1-6 Alkyl" or "C 1-3 "Alkyl" refers to methyl.

[0098] As used herein, "C 1-4 "Alkanediyl" includes methylene, 1,2-ethanediyl and their higher homologues.

[0099] As used herein, "O-alkyl" or "alkoxy" refers to an alkyl group of the indicated number of carbon atoms attached to an oxygen bridge. Thus, "O-alkyl" encompasses the definition of alkyl above. Preferably, O-alkyl is a straight or branched chain O-C alkyl. 1-6 Alkyl group, OC 1-4 Alkyl group, OC 1-3 Alkyl group or OC 1-2It refers to an alkyl group, or OCH. Examples of suitable O-alkyl groups include, but are not limited to, methoxy, ethoxy, n-propoxy, i-propoxy, n-butoxy, s-butoxy, or t-butoxy. Preferred alkoxy groups include methoxy, ethoxy, and t-butoxy.

[0100] As used herein, the terms "haloalkyl" and "O-haloalkyl" refer to an alkyl or alkoxy group in which one or more (particularly, one to three) hydrogen atoms have been replaced by halogen atoms (particularly, fluorine or chlorine atoms). The haloalkoxy group is preferably a straight-chain or branched-chain haloalkoxy, more preferably a haloC 1-3 Alkoxy groups, even more preferably haloC 1-2 Alkoxy groups are, for example, OCF3, OCHF2, OCH2F, OCH2CH2F, OCH2CHF2 or OCH2CF3, in particular OCF3 or OCHF2. Haloalkyl groups are preferably linear or branched haloalkyl groups, more preferably haloC 1-3 alkyl group, more preferably haloC 1-2 It is preferably an alkyl group, such as CF, CHF, CHF, CHCHF, CHCHF, CHCF, or CH(CH)CF. More preferably, it is preferably a haloalkyl group, haloC 1-6 Alkyl, HaloC 1-4 Alkyl group, halo C 1-3 Any one of the alkyl groups refers to CF3, CHF2, CH(CH3)CF3, CH2CF3 or (CH3)2CF3.

[0101] As used herein, the term "alkylamino" refers to an alkyl group of the indicated number of carbon atoms substituted with at least one amino group, where the amino group is -NH or is further substituted with one or two alkyl groups. For example, C 1-4Alkylamino refers to butylamine, isobutylamine, tert-butylamine, butyl-NH(CH), isobutyl-NH(CH), tert-butyl-NH(CH), butyl-N(CH), isobutyl-N(CH), tert-butyl-N(CH), butyl-NH(C2H), isobutyl-NH(C2H), tert-butyl-NH(C2H), butyl-N(C2H), isobutyl-N(C2H), tert-butyl-N(C2H), butyl-N(C2H)(CH), isobutyl-N(C2H)(CH), tert-butyl-N(C2H)(CH), and the like. As used herein, "OC 1-4 "Alkylamino" is the C bonded to the oxygen bridge 1-4 represents alkylamino.

[0102] As used herein, "NH-alkyl" refers to an alkyl group of the indicated number of carbon atoms attached to an NH bridge. Preferably, the NH-alkyl is a linear or branched NHC 1-6 Alkyl group, NHC 1-4 Alkyl group, NHC 1-3 Alkyl group or NHC 1-2 It refers to an alkyl group or NHCH3.

[0103] Similarly, "N(alkyl)2" represents two alkyl groups of the indicated number of carbon atoms attached through a nitrogen bridge.

[0104] As used herein, "S-alkyl" refers to an alkyl group of the indicated number of carbon atoms attached to a sulfur bridge. Thus, "S-alkyl" encompasses the definition of alkyl above. Preferably, S-alkyl is a straight or branched chain S-alkyl. 1-6 Alkyl group, SC 1-4 Alkyl group, SC 1-3 Alkyl group or SC 1-2The term "S-alkyl" refers to an alkyl group, or SCH. Examples of suitable S-alkyl groups include, but are not limited to, thiomethyl, thioethyl, thiopropyl, thio-i-propyl, thio-n-butyl, thio-s-butyl, or thio-t-butyl. Preferred S-alkyl groups include thiomethyl, thioethyl, and thiopropyl.

[0105] As used herein, the term "aryl" refers to a monocyclic or polycyclic aromatic ring containing carbon and hydrogen atoms. If necessary, such an aromatic ring may contain one or more heteroatoms, in which case it is also referred to as a "heteroaryl" or a "heteroaromatic ring." Illustrative examples of heteroaryl groups of the present invention include 5- or 6-membered heteroaryls such as thiophene, oxazole, oxadiazole, thiazole, thiadiazole, imidazole, pyrazole, pyrimidine, pyrazine, and pyridine. A preferred aryl of the present invention is phenyl. A preferred heteroaryl of the present invention is pyridyl. More preferred 5-membered heteroaryl rings are oxadiazole and oxazole. The oxadiazole is preferably substituted with one methyl group.

[0106] As used herein, the term "cycloalkyl" refers to a saturated cyclic hydrocarbon (cycloalkyl) having 3, 4, 5, or more carbon atoms and is a generic term for cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, and the like. The term "cycloalkyl" also refers to polycyclic saturated ring systems such as decahydronaphthalene, octahydro-1H-indene, and adamantane. The saturated ring may contain one or more heteroatoms (also referred to as "heterocyclyl" or "heterocycle" or "heterocycloalkyl") such that at least one carbon atom is replaced by a heteroatom selected from N, O, and S, particularly N and O. Preferably, the cycloalkyl is cyclohexyl, more preferably cyclopentyl. Preferably, the heterocycloalkyl is a cyclic amine such as piperidine, pyrrolidine, morpholine, or piperazine. More preferably, the heterocycloalkyl is tetrahydrofurane or tetrahydropyrane.

[0107] As used herein, the term "halogen" refers to fluorine, chlorine, bromine and iodine, of which fluorine, chlorine and bromine are preferred.

[0108] The compounds of the present invention may have asymmetric centers, chiral axes, and chiral planes (as described in E.L. Eliel and S.H. Wilen, Stereochemistry of Carbon Compounds, John Wiley & Sons, New York, 1994, pages 1119-1190) and may exist as racemates, racemic mixtures, and individual diastereomers, along with all possible isomers and mixtures thereof, including optical isomers; all such stereoisomers are included in the present invention. Compounds described in the present invention that contain olefinic double bonds include E and Z geometric isomers (cis-trans isomers) unless otherwise specified. The present invention also includes all salt forms, polymorphs, hydrates, and solvates.

[0109] The term "polymorph" refers to different crystalline structures of the compounds of the present invention. Polymorphs include, but are not limited to, crystalline forms (and amorphous materials) and all crystal lattice forms. The salts of the present invention may be crystalline and may exist as more than one polymorph.

[0110] The present invention also encompasses solvates, hydrates, and anhydrous forms of the salts or free compounds. The solvent contained in the solvate is not particularly limited and may be any pharmaceutically acceptable solvent. Examples include water and C 1-4 Alcohols (such as methanol or ethanol) are included.

[0111] "Pharmaceutically acceptable salts" are defined as derivatives of the disclosed compounds, in which the parent compound is modified by making acid or base salts thereof. Examples of pharmaceutically acceptable salts include, but are not limited to, mineral or organic acid salts of basic residues such as amines, alkali or organic salts of acidic residues such as carboxylic acids, etc. Said pharmaceutically acceptable salts include the conventional non-toxic salts or quaternary ammonium salts of the parent compound formed, for example, from non-toxic inorganic or organic acids. For example, such conventional non-toxic salts include those derived from inorganic acids such as, but not limited to, hydrochloric acid, hydrobromic acid, sulfuric acid, sulfamic acid, phosphoric acid, nitric acid, and the like, and organic acids such as, but not limited to, acetic acid, propionic acid, succinic acid, glycolic acid, stearic acid, lactic acid, malic acid, tartaric acid, citric acid, ascorbic acid, pamoic acid, maleic acid, hydroxymaleic acid, phenylacetic acid, glutamic acid, benzoic acid, salicylic acid, sulfanilic acid, 2-acetoxybenzoic acid, fumaric acid, toluenesulfonic acid, methanesulfonic acid, ethanedisulfonic acid, oxalic acid, isethionic acid, and the like. The pharmaceutically acceptable salts of the present invention can be synthesized from the parent compound which contains a basic or acidic moiety by conventional chemical methods. Generally, such salts can be prepared by reacting the free acid or base form of these compounds with a stoichiometric amount of an appropriate base or acid in water, an organic solvent, or a mixture of water and an organic solvent. Organic solvents include, but are not limited to, non-aqueous solvents such as ether, ethyl acetate, ethanol, isopropanol, or acetonitrile. A list of suitable salts can be found in Remington's Pharmaceutical Sciences, 18th ed., Mack Publishing Company, Easton, PA, 1990, p. 1445, the disclosure of which is incorporated herein by reference.

[0112] "Pharmaceutically acceptable" is defined as those compounds, materials, compositions and / or dosage forms that are, within the scope of sound medical judgment, suitable for use in contact with the tissues of human beings and animals without undue toxicity, irritation, allergic response, or other problem or complication commensurate with a reasonable benefit / risk ratio.

[0113] The compounds of the present invention are useful in a variety of applications for human and animal health. They are small molecules capable of modulating or eliminating mutant PrP activity. More specifically, they suppress the spontaneous cytotoxicity of mutant PrP(Δ105-125). Furthermore, because mutant PrP molecules sensitize cells to the cytotoxic effects of certain antibiotics, such as G418 and Zeocin, suppression of this antibiotic hypersensitivity phenotype was used as a cellular readout to screen small molecule libraries in the DBCA assay. Using this approach, compounds of the present invention were found to exhibit at least 30% activity relative to a reference compound, preferably greater than 60%, and even more preferably greater than 100% activity relative to the reference compound in suppressing the neurodegenerative phenotype.

[0114] The compounds of the present invention may indirectly modulate the farnesoid X receptor (FXR)-mediated signaling pathway. The farnesoid X receptor (FXR) is a nuclear receptor for bile acids. Ligand-activated FXR regulates gene transcription, allowing feedback control of bile acid synthesis and secretion. Under physiological conditions, activation of FXR is the primary mechanism for suppressing bile acid synthesis in both the liver and intestine by directly inducing target genes, including the small heterodimer partner (SHP / Shp, encoded by the NR0B2 / Nr0b2 gene) and fibroblast growth factor (Fgf) 15 (FGF19 in humans), which then inhibits CYP7A1 / Cyp7a1 and CYP8B1 / Cyp8b1 gene transcription or activates signaling pathways to inhibit said gene transcription. 36In the present invention, we also discovered that the FXR agonist WAY-362450, like SM231, potently rescues mutant PrP toxicity. Furthermore, SM231 significantly promoted FXR transcriptional activity in mouse primary hepatocytes, whereas SM derivatives do not act as direct FXR receptor agonists (data not shown). These findings suggest that indirect modulation of FXR activity may be involved in the mechanism of action of the compounds of the present invention.

[0115] The compounds of the present invention are used for the treatment of immune disorders. As used herein, "immune disorders" refers to autoimmune diseases or immune system disorders. In a preferred embodiment of the present invention, the immune disorders refer to autoimmune colitis, inflammatory bowel disease, or Crohn's disease.

[0116] The compounds of the present invention can be administered orally or parenterally in a dosage range of 0.001 to 1000 mg per kg of mammalian (e.g., human) body weight (0.001 to 1000 mg / kg) per day, in single or divided doses. One dosage range is 0.01 to 500 mg / kg of body weight per day, orally, in single or divided doses. Another dosage range is 0.1 to 100 mg / kg of body weight per day, orally, in single or divided doses. For oral administration, the compositions are provided in tablet or capsule form containing 1.0 to 500 milligrams of active ingredient, particularly 1, 5, 10, 15, 20, 25, 50, 75, 100, 150, 200, 250, 300, 400, and 500 milligrams, allowing for symptomatic adjustment of the dosage to the subject being treated. The specific dose level and frequency of administration for any particular subject will vary and will depend upon a variety of factors, including the activity of the particular compound used, the metabolic stability and duration of action of that compound, age, body weight, general health, sex, diet, mode and time of administration, rate of excretion, drug combination, severity of the particular condition, and the host undergoing therapy. [Brief explanation of the drawings]

[0117] BRIEF DESCRIPTION OF THE DRAWINGS Embodiments and experiments illustrating the principles of the present invention will be discussed with reference to the following figures. [Figure 1] Figure 1. Chemical and biological characterization of SM3. The compound SM3 (designated LD24 in the original study) with the structure shown in panel (A) was previously identified in an HTS screen for its ability to inhibit ΔCR PrP-dependent hypersensitivity to cationic antibiotics in a dose-dependent manner. (B) Drug-based cellular assays (DBCAs) were performed as previously described. Briefly, HEK293 cells stably transfected with a toxic PrP mutant with a deletion in the central region of the protein (Δ105-125) were seeded in 24-well plates and incubated at 37°C for 48 hours in medium containing 500 μg / mL Zeocin. Cell death in response to co-treatment with SM3 (0.1–10 μM) was assessed by MTT assay. Data are expressed as a percentage of untreated cells. [Figure 2] Figure 2. Evaluation of the potency of SM3 derivatives. The graph shows the relative ability of each compound (tested at 1 μM) to inhibit ΔCR PrP-dependent hypersensitivity to cationic antibiotics. Data are expressed as the mean percentage of the parent compound SM3 (referred to as LD24 in the graph). Error bars represent standard deviation. Each compound was tested in at least three biologically independent replicates (n≧3). [Figure 3]Figure 3. Dose-response analysis of selected compounds. (A) Chemical structures of selected molecules are shown in the panels. (B) Dose-response analysis by DBCA was used to evaluate the anti-ΔCR PrP efficacy of various molecules. The graph shows quantification of the dose-dependent rescuing effect of each molecule. Mean values ​​were obtained from a minimum of three independent experiments (n ≥ 3) and expressed as a percentage of cell viability in untreated cells. Data were fitted to a sigmoidal function using a four-parameter logistic (4PL) nonlinear regression model to estimate the half-maximal inhibitory concentration (IC50). (C) The intrinsic toxicity of each compound was evaluated in naive HEK293 cells exposed to each molecule / concentration for 48 hours at 37°C. Cytotoxicity was assessed by MTT. Mean values ​​were obtained from a minimum of three independent experiments (n ≥ 3) and expressed as a percentage of cell viability in untreated cells. Data were fitted to an inverse sigmoidal function using the 4PL nonlinear regression model to estimate the median lethal dose (LD50). [Figure 4]Figure 4. SM231 rescues Aβ neurotoxicity. Primary hippocampal neurons were exposed to various concentrations of Aβ oligomers for short periods (10, 20, or 60 min) or to vehicle (VHC) control. We confirmed that the oligomers induced rapid, transient phosphorylation of Fyn kinase (results at 20 min are shown in panel A). Consistent with previous findings, this effect was prevented by co-treatment with the PrPC-directed compound TMPyP. Interestingly, co-incubation with SM231 completely abolished the Aβ effect and restored Fyn phosphorylation to normal levels. (B) Primary hippocampal neurons were incubated with Aβ oligomers (3 μM) or VHC control for 3 h. Consistent with previous reports, we observed a decrease in several post-synaptic markers (as indicated) as assessed by Western blotting of Triton X-insoluble fractions. Importantly, co-incubation with SM231 for 20 min prior to incubation with Aβ oligomers significantly rescued (restored or normalized) the levels of all postsynaptic markers. The level of a control protein (actin) was unaffected by either Aβ oligomers or SM231 (*p<0.05; **p<0.01 by Student's t-test). [Figure 5]Figure 5. Metabolic studies on SM231. (A) The MetaSite software suggested three regions of the molecule as major metabolic sites (red and yellow in the 3D structure, bold spheres indicated by open arrows in the 2D structure), with the methylene bridge predicted to be the most reactive (highlighted by red / blue circles and filled arrows in the 2D structure of SM231). (B) Docking experiments with the active site of CYP450 indicated that only the cyclohexyl moiety and C-3 (gray arrow) are accessible to hepatic metabolic enzymes and therefore metabolizable. (C) Incubation of SM231 with rat liver microsomes (RLM) for 4 h showed a t1 / 2 of 25 min (right panel C), and HPLC-MS analysis of the resulting mixture suggested four metabolites (MET1–4), confirming that the cyclohexyl and C-3 are the major metabolic sites, albeit to a lesser extent. [Figure 6] Figure 6. DBCA-based validation of newly developed derivatives. The chemical structure of each molecule is shown in the graph. Dose-response analysis with DBCA was used to evaluate the anti-ΔCR PrP efficacy of various molecules. The graph shows the quantification of the rescue effect versus dose of each molecule. Mean values ​​were obtained from a minimum of three independent experiments (n ≥ 3) and expressed as a percentage of cell viability in untreated cells. Data were fitted to a sigmoidal function using the 4PL nonlinear regression model to estimate the half maximal inhibitory concentration (IC50). [Figure 7]Figure 7. SM884 rescues prion strain-induced suppression of LTP. The bar graph shows quantification of the rescue effect on LTP induced by chronic SM884 administration to brain slices acutely treated with lysate from cells infected with the mouse-adapted human M1000 prion strain. Values ​​are expressed as mean + / - SEM and calculated as the percentage of LTP rescue relative to vehicle control. Statistically significant differences between SM884-treated and untreated slices were calculated by Student's t-test. M1000 vs. 884 0.03 μM + M1000 p = 0.0038 (**); M1000 vs. 884 0.1 μM + M1000 p = 0.0031 (**). M1000 n=4;884 0.03μM+M1000 n=4;884 0.1μM+M1000 n=5. [Figure 8] Figure 8. DC subsets express endogenous PrPc, and SM231-treated DC2 promote Treg cell development in DC-T cell cocultures. A. Sorted murine DC1 and DC2 cells were treated with TMP or SM231, followed by Western blot analysis to assess PrPc expression using a specific anti-PrPc antibody. Results shown are the mean ± standard deviation (SD) from two independent EAE experiments. *P<0.05; two-tailed Mann-Whitney test. B. CD4+LAP+FOXP3+ cell frequency in cultures of DCs (i.e., DC1 or DC2) preconditioned with TMP, SM231, or vehicle, treated with either specific PrPc siRNA or siRNA control, and cultured with naive CD4+ T cells for 5 days. Representative results of CD4+CD25+LAP+FOXP3+ cell frequency in T:DC2 cocultures (upper right quadrant). Representative results from one of three experiments. [Figure 9]Figure 9. Compounds SM888 and SM889 promote tolerogenic activity in cDC2. cDC2 treated overnight with various concentrations of PrP regulatory molecules or vehicle as a control were co-cultured with CFSE-labeled CD4+ T cells from the spleens of OT.II mice in the presence of various concentrations of OVA. After 3 days, proliferation was analyzed by FACS to assess the % of T cell proliferation responding to specific antigens. Data are presented as mean ± standard deviation. **P<0.01, ***P<0.001, ****P<0.0001, ANOVA followed by Bonferroni's multiple comparison test. [Figure 10A] Figure 10. Administration of the reference PrP-binding molecule TMP or the PrP-activating molecule SM231 ameliorates EAE severity. A. Scheme of EAE induction and treatment. [Figure 10B] Figure 10. Administration of the reference PrP-binding molecule TMP or the PrP-activating molecule SM231 ameliorates EAE severity. B. EAE clinical scores (±SEM) in mice on a C57BL / 6 background treated with various doses of SM231 (*P<0.05; **P<0.01; ***P<0.001; Student's t-test). [Figure 10C] Figure 10. Administration of the reference PrP-binding molecule TMP or the PrP-activating molecule SM231 ameliorates EAE severity. C. Representative staining of spinal cord sections from MOG 35-55 immunized mice treated with PBS or SM231, visualizing immune infiltration and demyelination 25 days after immunization in mice treated with PBS or SM231. [Figure 11]Figure 11. SM231 does not directly affect PrP. (A) SM231 does not alter the cell surface localization of PrP. HEK293 cells stably expressing EGFP-tagged PrP were grown on glass coverslips to ∼60% confluence and then treated with the indicated concentrations of SM231 or CPZ for 24 h. After fixation and washing, the specific green signal of EGFP-PrP was captured using an inverted microscope coupled with a high-resolution camera equipped with a 488 nm excitation filter. (B) SM231 does not alter PrP expression. HEK293 cells expressing WT PrP were treated with various (indicated) concentrations of SM231 for 48 h. Total PrP levels in whole-cell lysates were assessed by Western blot using the anti-PrP antibody D18. The photograph in the top panel shows a representative Western blot. The graph in the lower panel shows quantification of PrP levels obtained by densitometry analysis of four independent experiments (n = 4), with each value normalized to the corresponding Ponceau S-stained lane. Bars represent the mean (±SEM) and are expressed as a percentage of the levels in untreated cells. (C) SM231 does not bind to PrP. The interaction of porphyrin Fe(III)-TMPyP (abbreviated as TP), chlorpromazine (CPZ), or SM231 with recombinant PrP was assessed by DMR. Varying concentrations of each compound (0.1–1000 μM) were added to label-free microplate well surfaces (EnSpire-LFB HS microplates, Perkin Elmer) pre-immobilized with full-length human recombinant PrP or BSA. Measurements were performed before (baseline) and after (final) compound addition. The response (pm) was obtained by subtracting the baseline signal from the final signal. The output signal for each well was obtained by subtracting the signal from the protein-coated reference region from the signal from the uncoated region. The TP (blue dots) or CPZ (green dots) signals were fitted to a sigmoid function using the 4PL nonlinear regression model (blue and green lines); R2 = 0.99; p = 0.00061). Conversely, no binding was detected for SM231, suggesting that this compound exerts no effect by directly binding to PrP. [Figure 12] Figure 12. FXR agonists potently rescue mutant PrP toxicity. The ability of two FXR agonists was tested at various concentrations (as indicated) to rescue Zeocin hypersensitivity induced by expression of the murine ΔCR PrP molecule in HEK293 cells using DBCA. The bar graph shows quantification of the rescue effect for each molecule dose. Mean values ​​were obtained from a minimum of three independent cell culture preparations and are expressed as the percentage of cell viability rescue using the following formula: R = (TZ) / (UZ) (R: rescue effect; T: cell viability of compound-treated samples; Z: cell viability of zeocin-treated samples; U: cell viability of untreated samples). [Figure 13] Figure 13. FXR transcriptional activity in hepatocytes treated with SM231. Mouse hepatocytes were treated for 4 or 12 hours as indicated. FXR and Nr0b2 mRNA levels were assessed by RT-qPCR. mRNA levels in untreated cells were arbitrarily set to 1. DETAILED DESCRIPTION OF THE INVENTION

[0118] Materials and Methods Chemistry: Methods for making compounds of general formula (I) As used herein, the following abbreviations have the following meanings: If an abbreviation is not defined, it has its generally accepted meaning.

[0119] Abbreviation BOP: N-(benzenesulfonyl)-L-prolyl-LO-(1-pyrrolidinylcarbonyl)tyrosine sodium salt; DIPEA: N,N-diisopropylethylamine, DMF: N,N-dimethylformamide, DMSO: N,N-dimethyl sulfoxide, EtOAc: ethyl acetate, MeOH: methanol, TBTU: 2-(1H-benzotriazol-1-yl)-1,1,3,3-tetramethylaminium tetrafluoroborate, Pyr: pyridine.

[0120] Unless otherwise noted, reagents and solvents were purchased from standard commercial suppliers and used as received. HPLC-grade solvents used in HPLC analysis were purchased from Sigma-Aldrich, and all mobile phases were degassed by sonication for 10 minutes before use. Organic solutions were dried over anhydrous Na2SO4 and concentrated under low pressure using a rotary evaporator. All reactions were routinely monitored by thin-layer chromatography (TLC) on silica gel 60 F254 (Merck) and visualized using UV or iodine. Microwave-assisted reactions were performed using a Biotage Initiator 2.0 microwave reactor, and parameters were adjusted depending on the reaction, as shown in the following examples. Flash chromatography was performed on Merck silica gel 60 (mesh 230–400). Melting points were measured in capillary tubes (Büchi Electrotermal Model 9100) and are uncorrected. 1 H NMR spectra were recorded at 200 MHz or 400 MHz (Bruker Avance DRX-200 or 400, respectively). 13 C NMR spectra were recorded at 100 MHz (Bruker Avance DRX-400) and 2D 1 H NMR NOESY was performed in phase-sensitive mode. Chemical shifts are expressed in ppm (δ) relative to TMS. Spectra were acquired at 298 K. Data processing was performed with standard Bruker software XwinNMR, and the spectral data are consistent with the assigned structures. Yields are for purified products and were not optimized. The purity of all compounds was ≥95% as determined by LC / MS using an Agilent 1290 Infinity System machine equipped with a DAD detector from 190 to 640 nm. Purity was determined using a ZORBAX Eclipse Plus C18 (2.1 × 50 mm, 1.8 μm). mThe analysis was performed using a 100% HCl (particle size) column at 270.44 nm. The reversed-phase column was run with 0.1% formic acid in CH3CN (channel B) and 0.1% formic acid in water (channel A) at 0.3 mL / min, with a gradient of 0-100% for 20 min. The injection volume was 0.5 μL, and the column temperature was 50 °C. The purity of all compounds was ≥95% as determined by HPLC using a Waters System machine equipped with a UV detector. Purity was measured using a X Terra C18 (x mm, μ m The HRMS detection was based on negative polarity electrospray ionization (ESI) using an Agilent 1290 Infinity System equipped with an Agilent 6540UHD Accurate Mass Q-TOF MS detector.

[0121] The compounds of the present invention can be prepared using a series of chemical reactions well known to those skilled in the art, and the process for preparing said compounds is constituted as a whole and can be further exemplified. The processes further described are meant to be exemplary only and are not meant to limit the scope of the present invention in any way. In particular, the compounds of the present invention can be prepared according to the general procedures outlined in the following schemes 1, 2, 3 and 4. Alternative synthetic routes and analogous structures will be apparent to those skilled in the art of organic chemistry.

[0122] Scheme 1 illustrates a procedure useful for preparing heterocyclic compounds of formula (I) having a dibenzo[c,e][1,2]thiazine 5,5 dioxide scaffold, i.e., where A is phenyl, B is phenyl, Y is an SO group, and where W is carbonyl, Z is nitrogen, X4 and X5 are hydrogen, and n is as defined in general formula (I). The substituent Q can be selected from the groups described in general formula (I).

[0123] Scheme 1. Synthetic procedures for preparing intermediates and target compounds

[0124] [ka]

[0125] Reagents and conditions: (i) aniline, dry Pyr, dry CH2Cl2, 40°C; (ii) Raney-Ni, H flux, DMF, room temperature (rt) or SnCl·2H O, 8N HCl, reflux, (iii) NaOH, NaNO2, then concentrated HCl, 0°C; (iv) Cu powder, DMSO, room temperature, (v) BrCH2CO2Et, DIPEA, DMF, microwave, 80°C, or alkyl alcohol, PhP3, DEAD, ultrasound, 25°C; (vi) excess amine, microwave, 120°C, neat conditions; (vii) 10% NaOH aqueous solution / EtOH (1:1), reflux; (viii) (a) amine, TBTU, DIPEA, dry CH2Cl2, room temperature, or (b) SOCl2, reflux, then amine, dry DMF, room temperature, or (c) BOP, DIPEA, dry CH2Cl2, room temperature.

[0126] Coupling of the appropriate 2-nitrobenzenesulfonyl chloride of formula (1a) with unsubstituted or functionalized anilines in dry pyridine at 40 °C afforded the corresponding aryl 2-nitrobenzenesulfonamides of formula (2a) in high yields. The nitro group of the intermediate of formula (2a) was reduced by catalytic reduction using Raney Ni and H2 flux or SnCl2·2H2O under acidic conditions, depending on the substrate, to give the amino compound of formula (3a). This was followed by diazotization using NaNO2 and HCl. After the addition of NaOH, the diazo compound was converted in situ to the unstable intermediate of formula (4a). These latter intermediates were immediately isolated as crude products and converted to the intermediates of formula (5a) in moderate yields using Cu powder and DMSO as the solvent at room temperature. Compounds of formula (5a) were reacted with ethyl 2-bromoacetate in DMF using DIPEA as a scavenger at 50°C for 15 minutes under microwave irradiation to give compounds of formula (6a) in good yields. Some intermediates of formula (5a) were alkylated using the Mitsunobu reaction to give certain compounds of formula (6a). Some examples of intermediates of formula (6a) were treated with an excess of an amine, represented by substituent Q, using microwave irradiation at 120°C and neat conditions to give target compounds of formula (8a). In some cases, the intermediate of formula (7a) was treated with a mixture of 10% aqueous NaOH and EtOH (1:1 ratio) to give the corresponding carboxylic acid of formula (7a), which was then coupled with an arylamine or alkylamine represented by the substituent Q. Other examples of target compounds of formula (8a) were obtained using two different methods: either using a condensing agent such as TBTU in CHCl with DIPEA as a scavenger, or SOCl as a chlorinating agent followed by the addition of an amine. In some cases, disubstituted anilines were used to prepare the intermediate of formula (5a) as a mixture of regioisomers, which were used directly in the next reaction step to give certain compounds of formula (8a). The regioisomers were then separated into the final compounds by flash chromatography to give each pure regioisomer.

[0127] Scheme 2 shows a procedure useful for preparing heterocyclic compounds of formula (I) having a dibenzo[c,e][1,2]thiazine 5,5 dioxide scaffold, where A is phenyl, B is phenyl, Y is a SO group, W is absent, Z is nitrogen, X and X are hydrogen, and n is as defined in general formula (I). The substituent Q can be selected from the groups described in general formula (I).

[0128] Scheme 2. Synthetic procedures for preparing some target compounds

[0129] [ka]

[0130] Reagents and conditions: alkyl halide, DIPEA, microwave, DMF, 70°C.

[0131] Certain compounds of formula (5a) were alkylated with bromo / chloroalkyls, where Z was selected from the substituents shown in formula (I) and n=1, 2, 3, 4, using the appropriate dihalide and DIPEA as a scavenger under microwave irradiation at 80°C.

[0132] Scheme 3 shows the procedure for synthesizing compounds SM226 and SM230 starting from compound SM225, which was demethylated with BBr in CHCl at −60° C. The reaction was then maintained at −30° C. to give the hydroxyl derivative SM226 used as an intermediate, which was subsequently O-alkylated with (2-chloroethyl)dimethylamine hydrochloride and CsCO in DMF at 80° C. to give the target compound SM230.

[0133] Scheme 3. Synthetic procedure for preparing target compounds SM226 and SM230

[0134] [ka]

[0135] Reagents and conditions: (i) 1M BBr3 in CH2Cl2, dry CH2Cl2, -60°C to -30°C; (ii) ClCH2CH2N(Me)2·HCl, Cs2CO3, dry DMF, 80°C.

[0136] Scheme 4 shows an example of a compound of formula (I) where A is phenyl, B is 3-methyl-pyrazole, Y is a SO group, W is carbonyl, and the substituent Q can be selected from the groups described in formula (I).

[0137] Scheme 4. Synthetic procedure for preparing target compound SM879

[0138] [ka]

[0139] Reagents and conditions: (v) cyclohexylamine, TBTU, DIPEA, dry THF, room temperature; (vi)

[0140] Compound 11a reported in Korean Patent No. KR2011060653 was reacted with cyclohexylamine in the presence of DIPEA using TBTU as a condensing agent to give intermediate 12a in good yield, which was then concentrated with hydrazine monohydrate at 60 °C under neat conditions to give the target product SM879.

[0141] The following examples are provided for the purpose of illustrating the present invention and should not be construed in any way as limiting the scope of the present invention.

[0142] [Table 1]

[0143] [Table 2]

[0144] [Table 3]

[0145] [Table 4]

[0146] [Table 5]

[0147] [Table 6]

[0148] The following examples are compounds purchased by AMBINTER and tested for biological activity as is. The synthetic procedures shown in Schemes 1-3 can be readily adapted to prepare commercially available compounds whose synthesis has not yet been reported.

[0149] [Table 7]

[0150] [Table 8]

[0151] [Table 9]

[0152] [Table 10]

[0153] experiment General procedure for obtaining nitrobenzenesulfonamides of Formula 2a (Scheme 1): To a solution of commercial or synthetic 2-nitrobenzenesulfonyl chloride (1 equiv.) and the appropriate aniline (2 equiv.) in CHCl, pyridine (1 equiv.) was added in one portion, and the mixture was maintained at 30 °C for 2 h under magnetic stirring. After concentrating to one-third of its volume, the mixture was poured into ice water and acidified (pH = 3) with 2 N HCl. After digestion under magnetic stirring, a precipitate formed. After filtration, the crude was triturated with cyclohexane / EtOAc (8:2) and filtered again to give benzenesulfonamides of Formula 2a.

[0154] Example 1 2-Nitro-N-[3-(trifluoromethyl)phenyl]benzenesulfonamide: Following the general procedure above and using 3-trifluoromethylaniline, the compound was obtained in 93% yield as a red solid, melting point (mp) 132.6-132.7 °C. 1 H NMR (200MHz, acetone-d6): δ 9.40 (brs, 1H, NH), 8.10-7.75 (m, 4H, Ar-H), 7.60-7.30 (m, 4H, Ar-H).

[0155] Example 2 N-(3-chloro-4-fluorophenyl)-2-nitrobenzenesulfonamide: Following the general procedure above with 3-chloro-4-fluoroaniline, the compound was obtained in 90% yield as a red solid, mp 110.0-110.1 °C. 1 H NMR (200MHz, acetone-d6): δ 9.25 (brs, 1H, NH), 8.00-7.70 (m, 4H, Ar-H), 7.50-7.40 (m, 1H, Ar-H), 7.30-7.20 (m, 2H, Ar-H).

[0156] Example 3 N-(3,5-Dichlorophenyl)-2-nitrobenzenesulfonamide: Following the general procedure outlined above and using 3,4-dichloroaniline, the compound was obtained in 95% yield as a red solid, mp 128.0-129.0 °C. 1 H NMR (400MHz, acetone-d6):δ 9.55(brs,1H,NH),8.20(d,J=1.3 and 7.8Hz,1H,Ar-H),8.10-7.80(m,3H,Ar-H),7.40-7.35(m,2H,Ar-H),7.28(t,J=1.8Hz,1H,Ar-H).

[0157] Example 4 5-Fluoro-2-nitro-N-[4-(trifluoromethyl)phenyl]benzenesulfonamide (2a(Int-1)): Following the general procedure outlined above, the intermediate 5-fluoro-2-nitrobenzenesulfonyl chloride (prepared as reported by Buhr in WO 212110603) was reacted with commercial 3-trifluoromethylaniline to afford the compound in 86% yield as a red solid, mp 130-132 °C. 1 H NMR (400MHz, CDCl3): δ 7.30-7.40 (m, 3H, Ar-H), 7.50-7.60 (m, 3H, Ar-H and NH), 7.70 (dd, J = 2.8 and 7.7Hz, 1H, Ar-H), 7.90 (dd, J = 4.5 and 8.8Hz, 1H, Ar-H).

[0158] Example 5 N-(4-Bromophenyl)-2-nitrobenzenesulfonamide: The intermediate was prepared according to the procedure reported in Kurkin, A. et al., Tetrahedron: Asymmetry, 2009, 20, 1500-1505. The melting point and spectral data are consistent with those reported in the literature.

[0159] Example 6 N-(3-Bromophenyl)-2-nitrobenzenesulfonamide: The intermediate was prepared according to the procedure reported in Abramovitch, RA et al., J. Org. Chem. 1977, 42, 2914-2919. The melting point and spectral data are consistent with those reported in the literature.

[0160] Example 7 2-Nitro-N-[4-(trifluoromethyl)phenyl]benzenesulfonamide: The intermediate was prepared according to the procedure reported in Kang, JG et al., Biosci. Biotechnol. Biochem. 2002, 66, 2677-2682. The melting point and spectral data are consistent with those reported in the literature.

[0161] Example 8 N-[4-(methylthio)phenyl]-2-nitrobenzenesulfonamide: The intermediate was prepared according to the procedure reported in PCT WO 2007 / 003962 A2. The melting point and spectral data are consistent with those reported in the literature.

[0162] Example 9 Scheme 5. Synthetic procedure for preparing intermediates of formula 2a (Int-2)

[0163] [ka]

[0164] 5-Methoxy-2-nitro-N-[4-(trifluoromethyl)phenyl]benzenesulfonamide (2a(Int-2)). A stirred mixture of 5-fluoro-2-nitro-N-[4-(trifluoromethyl)phenyl]benzenesulfonamide 2a(Int-1) (1.00 g, 2.86 mmol) in 10% aqueous NaOH (20 mL) and MeOH (40 mL) was kept at room temperature for 3 h. The reaction mixture was poured into ice / water, and the precipitate that formed was filtered off to give the title compound as a white solid in 99% yield. Melting point: 142-144 °C. 1 H NMR(400MHz,CDCl3):δ 3.89(s,3H,OCH3),7.12(dd,J=2.7 and 8.9Hz,1H,H-4),7.37(d,J=8.4Hz,2H,H-2' and H-6'), 7.48(d,J=2.7Hz,1H,H-6),7.58(d,J=8.5Hz,2H,H-3' and H-5'),8.02(d,J=9.0Hz,1H,H-3).

[0165] General procedure for obtaining aminobenzenesulfonamides of formula 3a (Scheme 1): A stirred solution of the nitro derivative of formula (2a) (1 equivalent) in EtOH (150 mL) was hydrogenated over catalytic Raney nickel at room temperature and atmospheric pressure for 2.5 h. The mixture was then filtered through Celite, and the filtrate was evaporated to dryness to give the pure amino derivative by TLC (CHCl3 / MeOH 98:2).

[0166] Example 10 2-Amino-N-[3-(trifluoromethyl)phenyl]benzenesulfonamide: Following the general procedure outlined above, the compound was obtained in 96% yield as an off-white solid, mp 88.1-88.2°C (dec.). 1 H NMR (200MHz, acetone-d6):δ 9.50(bs,1H,NH),7.60-7.25(m,5H,Ar-H),7.25-7.15(m,1H,Ar-H),6.75(d,J=8.3Hz,1H,Ar-H),6.50(t,J=8.0Hz,1H,Ar-H),6.75(bs,2H,NH2).

[0167] Example 11 2-Amino-N-(3-chloro-4-fluorophenyl)benzenesulfonamide: Following the procedure outlined above, the compound was obtained in 95% yield as a grey solid, mp 102.1-102.2 °C. 1 H NMR (200MHz, acetone-d6):δ 9.15(bs,1H,NH),7.40(dd,J=1.6 and 8.0Hz,1H,Ar-H),7.25-7.00(m,4H,Ar-H),6.75(d,J=8.3Hz,1H,Ar-H),6.55(t,J=8.0Hz,1H,Ar-H),5.60(bs,2H,NH2).

[0168] Example 12 2-Amino-N-(3,5-dichlorophenyl)benzenesulfonamide: Following the procedure outlined above, the compound was obtained in 92% yield as a grey solid, mp 103.0-105.0 °C. 1 H NMR (400 MHz, acetone-d₆): δ 9.50 (brs, 1H, NH₆), 7.59 (dd, J = 1.5 and 7.9 Hz, 1H, Ar-H), 7.25 (dt, J = 1.5 and 7.0 Hz, 1H, Ar-H), 7.18–7.12 (m, 2H, Ar-H), 7.10 (t, J = 2.4 Hz, 1H, Ar-H), 6.85 (dd, J = 0.9 and 7.9 Hz, 1H, Ar-H), 6.68 (dt, J = 0.9 and 7.0 Hz, 1H, Ar-H), 5.65 (brs, 2H, NH₂).

[0169] Example 13 2-Amino-N-[4-(trifluoromethyl)phenyl]benzenesulfonamide: Following the procedure outlined above, the compound was obtained in 85% yield as a grey solid (reaction time 1.5 h), melting point 105.3-105.4 °C. 1 H NMR(200MHz,DMSO-d6):δ 10.75(bs,1H,NH),7.40-7.60(m,3H,Ar-H),7.20-7.00(m,3H,Ar-H),6.70(d,J=8.0Hz,1H,Ar-H),6.50(t,J=8.0Hz,1H,Ar-H),5.95(bs,2H,NH2).

[0170] Example 14 2-Amino-N-[4-(methylthio)phenyl]benzenesulfonamide: To a stirred suspension of the corresponding nitro derivative of formula 2a (0.20 g, 0.62 mmol) in 8 N HCl (9.0 mL), SnCl₂·2H₂O (0.42 g, 1.85 mmol) dissolved in 8 N HCl (2.0 mL) was added in one portion, and the mixture was refluxed for 2 h. 10% NaOH was added until a pH of 6 was reached, and the precipitate thus obtained was filtered and washed three times with CHCl₃ (3 × 15 mL). Fractions were collected, the solvent was dried, and evaporated to dryness to give the amino derivative (0.10 g, 50% yield) as a crude solid, which was used directly in the next reaction step. mp 94.1–94.3 °C. 1 H NMR(200MHz,CDCl3):δ 7.40(dd,J=1.5 and 8.0Hz,1H,Ar-H),7.27-7.20(m,1H,Ar-H),6.75(d,J=8.3Hz,1H,Ar-H),7.10 -6.90(m,2H,Ar-H),6.80-6.90(m,2H,Ar-H),6.75-6.55(m,3H,Ar-H and NH),4.75(bs,2H,NH2).

[0171] Example 15 2-Amino-5-fluoro-N-[4-(trifluoromethyl)phenyl]benzenesulfonamide: Following the procedure outlined above, the compound was obtained in 87% yield as a pale orange solid (reaction time 1 h, purification method: trituration with cyclohexane), melting point 115-117 °C. 1 H NMR(400MHz,CD Cl3):δ 4.60(bs,2H,NH2),6.70(dd,J=4.3 and 8.8Hz,1H,Ar-H),7.05(dt,J=2.9 and 8.7Hz,1H,Ar-H),7 .15(d,J=8.4Hz,2H,Ar-H),7.30(dd,J=2.9 and 7.9Hz,1H,Ar-H),7.45(d,J=8.4Hz,2H,Ar-H).

[0172] Example 16 2-Amino-N-(3-bromophenyl)benzenesulfonamide: The intermediate was prepared according to the procedure reported in Abramovitch, RA et al., J. Org. Chem. 1977, 42, 2914-2919. The melting point and spectral data are consistent with those reported in the literature.

[0173] Example 17 2-Amino-N-(4-methoxyphenyl)benzenesulfonamide: The intermediate was prepared according to the procedure reported by Ramirez-Martinez, JF et al., Molecules, 2013, 18, 894-913. The spectral data are consistent with those reported in the literature.

[0174] Example 18 2-Amino-N-(4-chlorophenyl)benzenesulfonamide: The intermediate was prepared according to the procedure reported by Ramirez-Martinez, JF et al., Molecules, 2013, 18, 894-913. The spectral data are consistent with those reported in the literature.

[0175] Example 19 2-Amino-N-(2-bromophenyl)benzenesulfonamide: The intermediate was prepared according to the procedure reported in Giannotti, D. et al., J. Med. Chem. 1991, 34, 1356-1362. The spectral data are consistent with those reported in the literature.

[0176] Example 20 2-Amino-N-(3-chlorophenyl)benzenesulfonamide: The intermediate was prepared according to the procedure reported in PCT WO 96 / 05185. The melting point and spectral data are consistent with those reported in the literature.

[0177] Example 21 2-Amino-N-(4-bromophenyl)benzenesulfonamide: The intermediate was prepared according to the procedure reported by Ramirez-Martinez, JF et al., Molecules, 2013, 18, 894-913. The spectral data are consistent with those reported in the literature.

[0178] Example 22 2-Amino-5-methoxy-N-[4-(trifluoromethyl)phenyl]benzenesulfonamide. Following the procedure outlined above, the compound was obtained in 73% yield as a brown solid (reaction time 12 hours, purification method: trituration with EtO). Melting point 150-152 °C. 1 H NMR(400MHz,CDCl3):δ 3.64(s,3H,OCH3),5.38(bs,2H,NH2),6.52(d,J=8.4Hz,1H,H-3),6.66-6.69(m,1H,H-4), 6.82 (d, J=8.0Hz, 2H, H-2' and H-6'), 7.15-7.21 (m, 3H, H-6, H-3' and H-5'), 7.97 (s, 1H, NH).

[0179] General procedure for obtaining 6H-dibenzo[c,e][1,2]thiazine 5,5-dioxides of Formula 5a (Scheme 1): Aminobenzenesulfonamide of Formula 3a (1 equivalent), NaOH (1.2 equivalents), and NaNO (1.2 equivalents) were mixed in water, and the resulting solution was added dropwise to 37% HCl (6 equivalents) and kept at 0 °C. The muddy mixture was mixed with a glass rod for 30 minutes, and the formation of the diazonium salt was confirmed by β-naphthol assay. The red mixture was then diluted with HO and treated with AcONa powder until pH = 5. The orange solid thus obtained was filtered and treated with cyclohexane to give the unstable crude solid of Formula 4a. Due to its high instability, this solid was immediately added portionwise to a stirred suspension of Cu (5% by mass) powder in DMSO. After 30 min, the reaction mixture was filtered through Celite to remove Cu powder, and the filtrate was poured into ice / water and acidified with 2N HCl to pH=4 to give a precipitate, which was filtered under vacuum to give the intermediate compound of formula 5a.

[0180] Example 23 9-Bromo-6H-dibenzo[c,e][1,2]thiazine 5,5-dioxide: Following the general procedure outlined above, starting from the corresponding amino-benzenesulfonamide of formula 3a, the compound was obtained in 70% yield as a brown solid, which was used directly in the next reaction step. Melting point: 229-231 °C. 1 H NMR (200 MHz, DMSO-d₆): δ 11.50 (bs, 1H, NH), 8.37 (d, J = 2.3 Hz, 1H, Ar-H), 8.25 (d, J = 7.6 Hz, 1H, Ar-H), 7.90 (dd, J = 1.3 and 7.7 Hz, 1H, Ar-H), 7.76 (dt, J = 1.4 and 7.5 Hz, 1H, Ar-H), 7.66 (dd, J = 1.1 and 7.5 Hz, 1H, Ar-H), 7.59 (dd, J = 2.1 and 8.6 Hz, 1H, Ar-H), 7.55–7.80 (m, 3H, H-2, H-3, and H-8), 7.10 (d, J = 8.6 Hz, 1H, H-7).

[0181] Example 24 9-Chloro-6H-dibenzo[c,e][1,2]thiazine 5,5-dioxide: Following the general procedure outlined above, starting from the corresponding amino-benzenesulfonamide of formula 3a, the compound was obtained in 26% yield as a brown solid after purification by flash column chromatography (CHCl3 / MeOH 98:2) (reaction time 2 h), mp 231.4-231.5 °C. 1 H NMR (200 MHz, DMSO-d): δ 11.50 (bs, 1H, NH), 8.29–8.25 (m, 2H, Ar-H), 7.89 (dd, J = 1.5 and 7.5 Hz, 1H, Ar-H), 7.75 (dt, J = 1.5 and 7.5 Hz, 1H, Ar-H), 7.62 (dt, J = 1.0 and 9.0 Hz, 1H, Ar-H), 7.48 (dd, J = 2.3 and 7.2 Hz, 1H, Ar-H).

[0182] Example 25 9-(Trifluoromethyl)-6H-dibenzo[c,e][1,2]thiazine 5,5-dioxide: Following the general procedure outlined above, starting from the corresponding amino-benzenesulfonamide of formula 3a, the compound was obtained in 66% yield as a brown solid (reaction time 1 h), melting point 235-237 °C. 1 H-NMR (200MHz, CDCl3): δ 8.22(brs,1H,Ar-H),8.03-7.98(m,2H,Ar-H),7.80-7.74(m,2H,Ar - H),7.64-7.57(m,2H,Ar-H),7.20(brs,1H,NH).

[0183] Example 26 9-(Methylthio)-6H-dibenzo[c,e][1,2]thiazine 5,5-dioxide: Following the general procedure outlined above, starting from the corresponding amino-benzenesulfonamide of formula 3a, the compound was obtained in 25% yield as a pale brown solid after purification by flash column chromatography (CHCl / MeOH 98:2) (reaction time 2 h), mp 211.4-211.6 °C. 1H-NMR(200MHz,DMSO-d6):δ 11.29(brs,1H,NH),8.25(d,J=7.9Hz,1H,Ar-H),8.00(brs,1H,Ar-H),7.87(d,J=7.7Hz,1H,Ar-H),7.85(t ,J=7.3Hz,1H,Ar-H),7.61(t,J=7.4Hz,1H,Ar-H),7.34(d,J=8.5Hz,1H,Ar-H),7.09(d,J=8.5Hz,1H,Ar-H).

[0184] Example 27 9-Methoxy-6H-dibenzo[c,e][1,2]thiazine 5,5-dioxide: Following the general procedure outlined above, starting from the corresponding amino-benzenesulfonamide of formula 3a, the compound was obtained after purification by flash column chromatography (CHCl3 / MeOH 97:3) in 41% yield as a pale brown solid (reaction time: 30 min), mp 198-202 °C. 1 H-NMR(200MHz,DMSO-d6)δ 10.97(brs,1H,NH),8.25(d,J=7.7Hz,1H,Ar-H),7.85(dd,J=1.4 and 7.7Hz,1H,Ar-H),7.75(dt, J=1.4 and 7.55Hz, 1H, Ar-H), 7.68-7.55 (m, 2H, Ar-H), 7.13-6.95 (m, 2H, Ar-H), 3.80 (s, 3H, CH3).

[0185] Example 28 7-Bromo-6H-dibenzo-[c,e][1,2]thiazine 5,5-dioxide: Following the general procedure outlined above, starting from the corresponding amino-benzenesulfonamide of formula 3a, the compound was obtained in 78% yield as a yellow solid, mp 188.2-188.4 °C (dec.). 1 H-NMR(400MHz,DMSO-d6):δ 10.80(brs,1H,NH),8.25-8.30(m,2H,Ar-H),7.95(d,J=7.7Hz,1H,Ar-H),7.85-7.80(m,2H,Ar-H),7.72(t,J=7.5Hz,1H,Ar-H),7.36(t,J=7.9 Hz ,1H,Ar-H).

[0186] Example 29 8,10-Dichloro-6H-dibenzo[c,e][1,2]thiazine 5,5-dioxide: Following the general procedure outlined above, starting from the corresponding amino-benzenesulfonamide of formula 3a, the compound was obtained in 77% yield as a yellow solid, mp 190.0-191.0 °C (dec.). 1 H-NMR (200 MHz, acetone-d₆): δ 10.25 (brs, 1H, NH), 8.58 (dd, J = 1.7 and 7.8 Hz, 1H, Ar-H), 7.95 (dd, J = 1.5 and 7.3 Hz, 1H, Ar-H), 7.83–7.65 (m, 2H, Ar-H), 7.48 (d, J = 2.1 Hz, 1H, Ar-H), 7.32 (J = 2.1 Hz, 1H, Ar-H).

[0187] Example 30 3-Fluoro-9-(trifluoromethyl)-6H-dibenzo[c,e][1,2]thiazine 5,5-dioxide: Following the general procedure outlined above, starting from the corresponding amino-benzenesulfonamide of formula 3a, the compound was obtained in 83% yield as a pale brown solid, mp 195-197 °C. 1 H NMR (400 MHz, DMSO-d6): δ 7.35 (d, J = 8.4 Hz, 1H, H-7), 7.70 (dt, J = 2.7 and 8.7 Hz, 1H, H-2), 7.80 (d, J = 8.5 Hz, 1H, H-8), 7.85 (dd, J = 2.6 and 7.6 Hz, 1H, H-4), 8.50 (dd, J = 4.8 and 8.9 Hz, 1H, H-1), 8.55 (s, 1H, H-10), 12.10 (bs, 1H, NH).

[0188] Example 31 8-Bromo-6H-dibenzo[c,e][1,2]thiazine 5,5-dioxide and 10-bromo-6H-dibenzo[c,e][1,2]thiazine 5,5-dioxide: Following the general procedure shown above, starting from the corresponding amino-benzenesulfonamide of formula 3a, a mixture of two regioisomers was obtained that was difficult to separate, and this crude product was used in the next reaction step without further purification.

[0189] Example 32 8-Chloro-6H-dibenzo[c,e][1,2]thiazine 5,5-dioxide and 10-chloro-6H-dibenzo[c,e][1,2]thiazine 5,5-dioxide: Following the general procedure shown above, starting from the corresponding amino-benzenesulfonamide of formula 3a, a mixture of two regioisomers was obtained that was difficult to separate, and this crude product was used in the next reaction step without further purification.

[0190] Example 33 8-(Trifluoromethyl)-6H-dibenzo[c,e][1,2]thiazine 5,5-dioxide and 10-(trifluoromethyl)-6H-dibenzo[c,e][1,2]thiazine 5,5-dioxide: Following the general procedure shown above, starting from the corresponding amino-benzenesulfonamide of formula 3a, a mixture of two regioisomers was obtained that was difficult to separate, and this crude product was used in the next reaction step without further purification.

[0191] Example 34 8-Chloro-9-fluoro-6H-dibenzo[c,e][1,2]thiazine 5,5-dioxide and 10-chloro-9-fluoro-6H-dibenzo[c,e][1,2]thiazine 5,5-dioxide: Following the general procedure shown above, starting from the corresponding amino-benzenesulfonamide of formula 3a, a mixture of two regioisomers was obtained that was difficult to separate, and this crude product was used in the next reaction step without further purification.

[0192] Example 35 3-Methoxy-9-(trifluoromethyl)-6H-dibenzo[c,e][1,2]thiazine 5,5-dioxide: Following the general procedure outlined above, starting from the corresponding amino-benzenesulfonamide of formula 3a, the compound was obtained in 40% yield as a brown solid, mp 198-200 °C. 1H NMR(400MHz,DMSO-d6):δ 3.84(s,3H,OCH3),7.30-7.43(m,3H,Ar-H),7.75(d,J=7.2Hz,1H,Ar-H),8.37(d,J=8.6Hz,1H,Ar-H),8.49(s,1H,Ar-H),11.78(s,1H,NH).

[0193] General procedure for obtaining 6H-dibenzo[c,e][1,2]thiazine 5,5-dioxide N-6 ethyl acetate of Formula 6a (Scheme 1): In a microwave reaction tube, a solution of the appropriate compound of Formula 5a (1 equivalent), ethyl bromoacetate (1 equivalent), and DIPEA (3 equivalents) in dry DMF (5 mL) was irradiated at 50 °C for 15 min using the following experimental parameters: 5 bar pressure, cooling off, FHT on, and very high solvent uptake. The pitchy mixture was poured into ice water and extracted three times with EtOAc. The combined organic layers were washed with brine, dried, and evaporated to dryness to give a crude slurry mass, which was triturated with EtOH to give a precipitate that was filtered to give the desired compound.

[0194] Example 36 Ethyl 2-(9-bromo-5,5-dioxide-6H-dibenzo[c,e][1,2]thiazin-6-yl)acetate: Following the general procedure outlined above, starting from the corresponding dibenzothiazine of formula 5a, the compound was obtained in 75% yield as a pink solid, mp 89-91 °C. 1 H-NMR(200MHz,DMSO-d6):δ 8.40(d,J=2.3Hz,1H,Ar-H),8.26(d,J=7.6Hz,1H,Ar-H),7.90-7.60(m,4H,Ar-H),7.40(d,J=8.6 Hz,1H,Ar-H),4.77(s,2H,NCH2),3.90(q,J=7.0Hz,2H,OCH2CH3),0.80(t,J=7.0Hz,3H,OCH2CH3).

[0195] Example 37 Ethyl (9-chloro-5,5-dioxide-6H-dibenzo[c,e][1,2]thiazin-6-yl)acetate: Following the general procedure outlined above, starting from the corresponding dibenzothiazine of formula 5a, the compound was obtained in 86% yield as a brown solid, mp 102.8-102.9 °C. 1 H-NMR(200MHz, CDCl3):δ 8.47-7.99(m,3H,Ar-H),7.64(dt,J=1.5 and 7.5Hz,1H,Ar-H),7.53(dt,J=1.2 and 7.7Hz,1H,Ar-H),7.34(dd,J=2.3 and 8.7Hz, 1H,Ar-H),7.17(d,J=8.7Hz,1H,Ar-H),4.59(s,2H,NCH2),3.96(q,J=7.2Hz,2H,OCH2CH3),1.00(t,J=7.2Hz,3H,OCH2CH3).

[0196] Example 38 Ethyl [5,5-dioxido-9-(trifluoromethyl)-6H-dibenzo[c,e][1,2]thiazin-6-yl]acetate: Following the general procedure outlined above, starting from the corresponding dibenzothiazine of formula 5a, the compound was obtained in 80% yield as a pale brown solid, mp 101-103 °C. 1 H-NMR(200MHz,DMSO-d6):δ 8.60(brs,1H,Ar-H),8.35(d,J=8.0Hz,1H,Ar-H),8.00-7.75(m,3H,Ar-H),7.74-7.65(m,2H ,Ar-H),4.91(s,2H,NCH2),3.92(q,J=7.4Hz,2H,OCH2CH3),0.92(t,J=7.4Hz,3H,OCH2CH3).

[0197] Example 39 Ethyl [9-(methylthio)-5,5-dioxide-6H-dibenzo[c,e][1,2]thiazin-6-yl]acetate: Following the general procedure outlined above, starting from the corresponding dibenzothiazine of formula 5a, the compound was obtained in 73% yield as a yellowish solid, mp 103-105 °C. 1H-NMR(200MHz,DMSO-d6):δ 8.02-7.91(m,3H,Ar-H),7.75(t,J=7.9Hz,1H,Ar-H),7.61(t,J=7.4Hz,1H,Ar-H),7.45-7.35(m,1H,Ar-H),7.26-7.2 3(m,1H,Ar-H),4.66(s,2H,NCH2),4.05(q,J=6.9Hz,2H,OCH2CH3),2.57(s,3H,SCH3),1.07(t,J=6.9Hz,3H,OCH2CH3).

[0198] Example 40 Ethyl (9-methoxy-5,5-dioxide-6H-dibenzo[c,e][1,2]thiazin-6-yl)acetate: Following the general procedure outlined above, starting from the corresponding dibenzothiazine of formula 5a, the compound was obtained in 85% yield as a brown solid, mp 101-104 °C. 1 H-NMR(200MHz,DMSO-d6):δ 8.26(d,J=7.9Hz,1H,Ar-H),7.88-7.74(m,2H,Ar-H),7.70-7.62(m,2H,Ar-H),7.49(d,J=8.9Hz,1H,Ar-H),7.12(d d, J = 2.7 and 8.9Hz, 1H, Ar-H), 4.75 (s, 2H, NCH2), 3.94-3.77 (m, 5H, OCH3 and OCH2CH3), 0.90 (t, J = 7.0Hz, 3H, OCH2CH3).

[0199] Example 41 Ethyl (7-bromo-5,5-dioxide-6H-dibenzo[c,e][1,2]thiazin-6-yl)acetate: Following the general procedure outlined above, starting from the corresponding dibenzothiazine of formula 5a, the compound was obtained in 50% yield as a pink solid after crystallization from EtOH, mp 169-171 °C. 1 H-NMR(400MHz,CDCl3)δ 7.96-7.90(m,3H,Ar-H),7.75-7.67(m,2H,Ar-H),7.58(t,J=7.8 Hz,1H,Ar-H),7.30(t,J=7.9Hz,1H,Ar-H),4.73(s,2H,NCH2),3.80(q,J=7.1Hz,2H,OCH2CH3),1.00(t,J=7.1Hz,3H,OCH2CH3).

[0200] Example 42 Ethyl (8,10-dichloro-5,5-dioxide-6H-dibenzo[c,e][1,2]thiazin-6-yl)acetate: Obtained in 90% yield as a pink solid, mp 172-173 °C, starting from the corresponding dibenzothiazine of formula 5a according to the general procedure outlined above. 1 H-NMR(200MHz,CDCl3)δ 8.49(dd,J=1.3 and 7.9Hz,1H,Ar-H),7.90(dd,J=1.8 and 7.6Hz,1H,Ar-H),7.70-7.55(m,2H,Ar-H),7.10(d ,J=2.1Hz,1H,Ar-H),4.51(s,2H,NCH2),4.02(q,J=7.2Hz,2H,OCH2CH3),1.05(t,J=7.2Hz,3H,OCH2CH3).

[0201] Example 43 Ethyl [3-fluoro-5,5-dioxide-9-(trifluoromethyl)-6H-dibenzo[c,e][1,2]thiazin-6-yl]acetate: Obtained according to the general procedure outlined above, starting from the corresponding dibenzothiazine of formula 5a, in 85% yield as a pale brown solid, mp 175-177 °C. 1 H NMR(400MHz,DMSO-d6):δ 1.10(t,J=7.2Hz,3H,OCH2CH3),4.10(q,J=7.2Hz,2H,OCH2CH3),4.75(s,1H,NCH2),7.35(d,J=8.5Hz,1H,H-7),7.4 5 (dt, J=2.7 and 8.3Hz, 1H, H-2), 7.60-7.70 (m, 2H, H-4 and H-8), 8.00 (dd, J = 4.6 and 8.8Hz, 1H, H-1), 8.20 (s, 1H, H-10).

[0202] Example 44 Ethyl [3-methoxy-5,5-dioxide-9-(trifluoromethyl)-6H-dibenzo[c,e][1,2]thiazin-6-yl]acetate: Following the general procedure outlined above, starting from the corresponding dibenzothiazine of formula 5a, the compound was obtained in 86% yield as a pink solid, mp 190-192 °C. 1 H NMR(400MHz,DMSO-d6):δ 1.03(t,J=7.0Hz,3H,OCH2CH3),3.95(s,3H,OCH3),3.98(q,J=7.4Hz,2H,OCH2CH3),4.93(s,2H,NCH2),7.42-7.45(m, 2H,H-2 and H-4), 7.73(d,J=8.6Hz,1H,H-7),7.85(d,J=8.4Hz,1H,H-8),8.38(d,J=8.4Hz,1H,H-1),8.54(s,1H,H-10).

[0203] Example 45 N-[2-(9-Bromo-5,5-dioxide-6H-dibenzo[c,e][1,2]thiazin-6-yl)ethyl]cyclohexanamine (SM9). In a microwave reaction tube, a solution of the appropriate compound of formula 5a (0.6 g, 1.9 mmol), N-(2-chloroethyl)cyclohexanamine (0.31 g, 1.9 mmol), and DIPEA (0.66 mL, 3.8 mmol) in dry DMF (4 mL) was irradiated at 70 °C for 60 min under the following experimental parameters: pressure 5 bar, cooling off, FHT on, very high solvent absorption. The residue was poured into ice water, acidified to pH 3 with 2 N HCl, and extracted with EtOAc (3 × 20 mL). The combined organic layers were washed with brine, dried and evaporated to dryness to give a crude slurry mass which was purified by flash column chromatography eluting with CHCl3 / MeOH (97:3) to give the target compound SM9 (0.65 g, 79%) as a low-melting pale brown solid. 1H NMR(400MHz,CDCl3):δ 8.13(d,J=2.2Hz,1H,Ar-H),7.99(dd,J=1.1 and 7.7Hz,1H,H-4),7.92(d,J=7.2Hz,1H,Ar -H),7.73(dt,J=1.3 and 7.8Hz,1H,Ar-H),7.64-7.58(m,2H,Ar-H),7.40(d,J=8.7Hz,1H,A r-H),4.00(t,J=6.7Hz,2H,SO2NCH2),2.85(t,J=6.7Hz,2H,NCH2),2.40-2.25(m,1H,Cy -CH),1.75-1.50(m,4H,Cy-CH2),1.25-1.00(m,4H,Cy-CH2),1.00-0.80(m,2H,Cy-CH2). 13 C NMR(100MHz,CDCl3):δ 137.58,135.22,133.05,132.42,131.16,128.95,128.45,127.15,125. 68,123.56,122.51,118.59,56.17,49.17,44.52,32.97,25.87,24.72. HRMS(ESI) C 20 H 23 BrN2O2S [M + +H] + Calculated value (calcd.): 435.0739, found value: 435.0735. LC-MS: Retention time (ret. time) 4.075.

[0204] Example 46 (R,S)-3-(9-bromo-5,5-dioxide-6H-dibenzo[c,e][1,2]thiazin-6-yl)-1-cyclohexylpyrrolidin-2-one (SM11). Following the procedure described above for compound SM9, 3-bromo-1-cyclohexyl-2-pyrrolidinone was used. 37 The title compound was purified by flash column chromatography eluting with cyclohexane / EtOAc (6:4) followed by trituration with petroleum ether / Et2O to give the target racemic compound SM11 in 70% yield as a white solid, mp 177-179 °C. 1H NMR(400MHz,DMSO-d6):δ 8.42(d,J=2.1Hz,1H,Ar-H),8.28(d,J=7.8Hz,1H,Ar-H),7.91(d,J=7.7Hz,1H,Ar-H),7.85(t,J= 8.6Hz,1H,Ar-H),7.75-7.70(m,2H,Ar-H),7.30(d,J=8.6Hz,1H,H-7),7.37(d,J=8.6Hz,1H,Ar-H) ,4 0.90 (t, J = 9.4 Hz, 2-pyrrolidone-CH), 3.75-3.55 (m, 1H, Cy-CH), 3.20-3.00 (m, 2H, 2-pyrrolidone-CH), 2.10-2.00 (m, 1H, 2-pyrrolidone-CH), 1.75-1.48 (m, 6H, Cy-CH2 and 2-pyrrolidone-CH), 1.40-1.00 (m, 5H, Cy-CH2). 13 C NMR(100MHz,DMSO-d6):δ 168.65,136.83,135.87,133.56,133.48,131.05,130.15,129.38,128.95,127 .40,126.72,122.36,119.96,62.56,51.37,25.45,25.38,25.33,25.19,22.83. HRMS(ESI) C 22 H 23 BrN2O3S:[M + +H] + Calculated: 475.0692, Found: 475.03691. LC-MS: Retention time 6.015.

[0205] Example 47 Methyl 3-(9-bromo-5,5-dioxide-6H-dibenzo[c,e][1,2]thiazin-6-yl)propanoate: To a solution of the appropriate compound of Formula 5a (0.300 g, 0.92 mmol) in dry THF (12 mL) was added commercial methyl 3-hydroxypropanoate (0.12 mL, 1.3 mmol) and PPh3 (0.33 g, 1.3 mmol), and the solution was sonicated at 25 °C for 7 min. DEAD (0.20 mL, 1.3 mmol) was then added dropwise, and the solution was sonicated at 25 °C for 18 h (approximately 70% conversion was observed by TLC). The mixture was concentrated under reduced pressure, poured into ice water, basified to pH 10 with 10% aqueous NaOH to remove remaining starting material, and extracted with EtOAc (3 × 20 mL). The combined organic layers were washed with brine, dried, and evaporated to dryness. The resulting brown oil was purified by column chromatography (petroleum ether / EtOAc 7:3) followed by trituration with EtO to give the desired title compound of formula 6a (0.100 g, 30%) as a white solid, mp 101-103 °C. 1 H NMR(200MHz,CDCl3):δ 8.10(d,J=2.3Hz,1H,Ar-H),7.94-7.83(m,2H,Ar-H),7.70(dt,J=1.5 and 7.4Hz,2H,Ar-H),7.60-7.50(m,2H,A r-H),7.28(d,J=8.8Hz,1H,Ar-H),4.10(t,J=7.4Hz,2H,NCH2),3.50(s,3H,CH3),2.50(t,J=7.4Hz,2H,CH2).

[0206] General procedure for the direct amidation of compounds of formula 6a with cyclohexylamine to give target compounds of formula 8a (Scheme 1): A tube containing the appropriate dibenzothiazine ethyl acetate or a mixture of the intermediate of general formula 6a in Example 44 (1 equivalent) and cyclohexylamine (4 equivalents) was irradiated in a microwave oven at 120°C for 4 hours with the following experimental parameters: pressure 5 bar, cooling off, FHT on, solvent absorption normal. The residue was dissolved in ice water. It was poured and acidified with 2N HCl to pH 3. The resulting precipitate was filtered and crystallized with EtOH to give the target compound of formula 8a.

[0207] Example 48 2-(9-Bromo-5,5-dioxide-6H-dibenzo[c,e][1,2]thiazin-6-yl)-N-cyclohexylacetamide (SM3): Following the general procedure described above, the title compound was obtained in 62% yield as a white solid, mp 211-213 °C. 1 H NMR(400MHz,CDCl3):δ 8.20(brs,1H,Ar-H),8.00(d,J=7.8Hz,1H,Ar-H),7.87(d,J=7.8Hz,1H,Ar-H),7.75(t,J=7. 5Hz,1H,Ar-H),7.73(t,J=7.4Hz,1H,Ar-H),7.57(d,J=7.4Hz,1H,Ar-H),7.12(d,J=8.7Hz,1 H,H-7),6.54(d,J=7.0Hz,1H,NH),4.42(s,2H,NCH2),3.90-3.75(m,1H,Cy-CH),1.95-1.75( m,2H,Cy-CH),1.65-1.50(m,3H,Cy-CH),1.45-1.25(m,2H,Cy-CH),1.25-1.05(m,3H,Cy-CH). 13 C NMR(100MHz,DMSO-d6):δ 166.29,137.11,134.09,133.49,133.12,131.05,129.18,128.57,125. 91,125.59,122.56,121.09,118.62,51.64,48.45,32.45,25.35,24.35. HRMS(ESI) m / z[M+H] + C 20 H 21 Calculated for BrN2O3S: 448.0535, found: 448.0456. LC-MS: retention time 5.754.

[0208] Example 49 2-(9-Bromo-5,5-dioxide-6H-dibenzo[c,e][1,2]thiazin-6-yl)-N-cyclopentylacetamide (SM4): Following the general procedure described above, the title compound was obtained after crystallization from EtOH in 43% yield as a white solid, mp 192-194 °C. 1 H NMR(400MHz,CDCl3):δ 8.19(d,J=2.1Hz,1H,Ar-H),8.04(d,J=7.9Hz,1H,Ar-H),7.99(d,J=7.9Hz,1H,Ar-H),7. 80(dt,J=1.2 and 7.6Hz,1H,Ar-H),7.69-7.61(m,2H,Ar-H),7.15(d,J=8.7Hz,1H,Ar-H),6 .62 (d, J = 7.5 Hz, 1H, NH), 4.46 (s, 2H, NCH2), 4.31-4.24 (m, 1H, cyclopentyl-CH), 2.00-1.92 (m, 2H, cyclopentyl-CH2), 1.70-1.55 (m, 4H, cyclopentyl-CH2), 1.45-1.30 (m, 2H, cyclopentyl-CH2). 13 C NMR(100MHz,CDCl3):δ166.76,137.03,134.04,133.49,133.14,131.00,129.20 ,128.57,125.90,125.53,122.53,121.04,118.61,51.60,51.49,32.80,23.45. HRMS(ESI) m / z[M+H] + C 19 H 19 Calculated for BrN2O3S: 435.0379, found: 435.03733. LC-MS: retention time 5.434.

[0209] Example 50 2-(9-Bromo-5,5-dioxide-6H-dibenzo[c,e][1,2]thiazin-6-yl)-N-cycloheptylacetamide (SM5): Following the general procedure described above, the title compound was obtained after crystallization from EtOH in 51% yield as a white solid, mp 208-210 °C. 1H NMR(400MHz,CDCl3):δ 8.18(d,J=1.8Hz,1H,Ar-H),8.03(d,J=7.4Hz,1H,Ar-H),7.98(d,J=7.9Hz,1H,Ar-H),7.80(t,J=7.6Hz,1H,H-2),7.70-7.60( m,2H,Ar-H),7.15(d,J=8.7Hz,1H,Ar-H),6.60(d,J=7.5Hz,1H,NH),4.45(s,2H,NCH2),4.20-3.95(m,1H,cycloheptyl-CH),1.90-1 . 77 (m, 2H, cycloheptyl-CH2), 1.70-1.30 (m, 10H, cycloheptyl-CH2). 13 C NMR(100MHz,CDCl3):δ 166.01,137.08,134.08,133.47,133.13,131.04,129.18,128.57,125. 92,125.55,122.55,121.04,118.60,51.60,50.63,34.59,27.84,23.72. HRMS(ESI) m / z[M+H] + C 21 H 23 Calculated for BrN2O3S: 463.0689, found: 463.0688. LC-MS: retention time 6.018.

[0210] Example 51 3-(9-Bromo-5,5-dioxide-6H-dibenzo[c,e][1,2]thiazin-6-yl)-N-cyclohexylpropanamide (SM10). Following the general procedure described above, the title compound was obtained after purification by flash column chromatography (cyclohexane / EtOAc 7:3) in 34% yield as a white solid. Melting point 114-116 °C. 1H NMR (400 MHz, CDCl): δ 8.13(brs,1H,Ar-H),8.00(d,J=7.6Hz,1H,Ar-H),7.90(d,J=7.7Hz,1H,Ar-H),7. 75(t,J=7.7Hz,1H,Ar-H),7.65-7.60(m,2H,Ar-H),7.42(d,J=8.7Hz,1H,Ar-H),5 .55(d,J=6.4Hz,1H,NH),4.15(t,J=6.6Hz,2H,NCH2),3.75-3.60(m,1H,Cy-CH),2 .60(t,J=6.6Hz,2H,CH2),1.85-1.53(m,5H,Cy-CH2),1.50-1.10(m,5H,Cy-CH2). 13 C NMR(100MHz,CDCl3):δ 168.33,137.37,134.19,132.91,132.21,130.81,128.46,127.79,126.07, 125.26,123.13,122.13,118.23,47.96,45.98,36.49,32.36,24.90,24.18. HRMS(ESI) C 21 H 23 BrN2O3S [M + +H] + Calculated: 463.0689, Found: 463.0693. LC-MS: Retention time 4.772.

[0211] Example 52 2-(9-Chloro-5,5-dioxide-6H-dibenzo[c,e][1,2]thiazin-6-yl)-N-cyclohexylacetamide (SM254): Following the general procedure described above, the title compound was obtained after crystallization from EtOH in 60% yield as a white solid, mp 218-220 °C. 1H NMR(400MHz,CDCl3):δ 8.02-7.95(m,2H,Ar-H),7.92(d,J=8.1Hz,1H,Ar-H),7.75(dt,J=1.2 and 7.7Hz,1H,Ar-H),7 .63(dt,J=0.8 and 8.0Hz,1H,Ar-H),7.43(dd,J=2.2 and 8.7Hz,1H,Ar-H),7.18(d,J=8.7Hz,1H, H-7),6.53(d,J=7.0Hz,1H,NH),4.42(s,2H,NCH2),3.90-3.75(m,1H,Cy-CH),1.90-1.75(m ,2H,Cy-CH),1.65-1.50(m,4H,Cy-CH),1.40-1.25(m,2H,Cy-CH),1.25-1.05(m,2H,Cy-CH). 13 C NMR(100MHz,CDCl3):δ 166.29,136.69,134.16,133.08,131.17,131.11,130.58,129.16,125. 88,125.61,125.32,122.58,120.97,51.75,48.45,32.45,25.34,24.32. HRMS(ESI) m / z[M+H] + C 20 H 21 Calculated for ClN2O3S: 405.1039, found: 404.1032. LC-MS: Retention time 6.492 min.

[0212] Example 53 N-Cyclohexyl-2-[5,5-dioxide-9-(trifluoromethyl)-6H-dibenzo[c,e][1,2]thiazin-6-yl]acetamide (SM231): Following the general procedure described above, the title compound was obtained after purification by flash column chromatography (cyclohexane / EtOAc 7:3) in 70% yield as a white solid, mp 225-226 °C. 1H-NMR(200MHz, CDCl3):δ 8.29(brs,1H,Ar-H),8.03(d,J=7.5Hz,1H,Ar-H),7.81(d,J=7.8Hz,1H,Ar-H),7.75-7.65(m,2H,Ar-H),7.35(d,J=8.4Hz,1H,Ar-H),6.57(brs,1H,NH) ,4.52(s,2H,NCH2),3.85-3.75(m,1H,Cy-CH),1.85-1.75(m,2H,Cy-CH),1. 65-1.48(m,3H,Cy-CH),1.40-1.25(m,2H,Cy-CH),1.20-1.10(m,3H,Cy-CH). 13 C NMR(100MHz,CDCl3):δ 166.04,140.60,134.01,133.29,131.16,129.36,127.33(q,J C-F =33.1Hz,C-9), 127.32(d,J C-F =3.5Hz,C-10),126.02,123.86,123.63(q,J C-F =270.7Hz,CF3),122.96(d,J C-F =3.8Hz,C-8),119.49,51.25,48.55,32.43,25.32,24.33. HRMS(ESI) m / z[M+Na] + C 21 H 21 Calculated for F3N2O3S: 461.1118, found: 461.1124. LC-MS: Retention time 4.688 min.

[0213] Example 54 N-Cyclohexyl-2-[9-(methylthio)-5,5-dioxide-6H-dibenzo[c,e][1,2]thiazin-6-yl]acetamide (SM340): Following the general procedure described above, the title compound was obtained after crystallization from EtOH in 63% yield as a white solid, mp 178-180 °C. 1H-NMR(400MHz, CDCl3):δ 8.03-7.98(m,2H,Ar-H),7.92(d,J=1.6Hz,1H,Ar-H),7.78(t,J=7.6Hz,1H,Ar-H),7.64(t,J =7.4Hz,1H,Ar-H),7.39(dd,J=1.9 and 8.6Hz,1H,Ar-H),7.20(d,J=8.6Hz,1H,Ar-H),6.60(d,J =7.5Hz,1H,NH),4.44(s,2H,NCH2),3.90-3.75(m,1H,Cy-CH),2.58(s,3H,SCH3),1.90-1.80( m,2H,Cy-CH),1.75-1.50(m,3H,Cy-CH),1.40-1.25(m,2H,Cy-CH),1.20-1.10(m,3H,Cy-CH). 13 C-NMR(100MHz,CDCl3):δ 166.60,135.92,135.61,134.20,132.99,131.79,129.14,128.79,125.81, 124.44,124.07,122.59,120.25,51.84,48.39,32.46,25.35,24.35,16.44. HRMS(ESI) m / z[M+H] + C 21 H 24 Calculated for N2O3S2: 417.1309, found: 417.1305. LC-MS: Retention time 5.403 min.

[0214] Example 55 N-Cyclohexyl-2-(9-methoxy-5,5-dioxide-6H-dibenzo[c,e][1,2]thiazin-6-yl)acetamide (SM225): Following the general procedure described above, the title compound was obtained after purification by flash column chromatography (cyclohexane / EtOAc 7:3) in 45% yield as a white solid, mp 216-217 °C. 1H-NMR(400MHz, CDCl3):δ 7.99-7.93(m,2H,Ar-H),7.74(t,J=7.6Hz,1H,Ar-H),7.59(t,J=7.6Hz,1H,Ar-H),7.49(d ,J=2.6Hz,1H,Ar-H),7.20(d,J=8.9Hz,1H,Ar-H),7.02(dd,J=2.7 and 8.9Hz,1H,Ar-H),6.59 (d,J=7.8Hz,1H,NH),4.32(s,2H,NCH2),3.88-3.75(m,4H,OCH3,and Cy-CH),1.85-1.75(m, 2H,Cy-CH),1.65-1.50(m,3H,Cy-CH),1.35-1.25(m,2H,Cy-CH),1.20-1.10(m,3H,Cy-CH). 13 C-NMR(100MHz,CDCl3):δ 166.82,157.36,134.29,132.96,132.22,131.86,128.76,125.87,125.50, 122.84,121.68,116.50,110.77,55.78,52.75,48.27,32.52,25.39,24.43. HRMS(ESI) m / z[M+H] + C 21 H 24 Calculated for N2O4S: 401.1539, found: 401.1533. LC-MS: Retention time 5.544 min.

[0215] Example 56 2-(7-Bromo-5,5-dioxide-6H-dibenzo[c,e][1,2]thiazin-6-yl)-N-cyclohexylacetamide (SM227): Following the general procedure described above, the title compound was obtained after purification by flash column chromatography (cyclohexane / EtOAc 6:4) in 40% yield as a white solid, mp 159-160 °C. 1H-NMR (200 MHz, DMSO-d₆) δ 8.15 (d, 2H, H-4 and H-8), 7.70–7.85 (m, 4H, H-1, H-9, H-10 and NH₂), 7.55–7.70 (t, J = 7.4 Hz, 1H, H-2), 7.40 (t, J = 7.9 Hz, 1H, H-3), 4.40 (brs, 2H, NCH₂), 3.00–3.10 (m, 1H, cyclohexyl CH₂), 0.60–1.60 (m, 10H, cyclohexyl CH₂). 13 C-NMR(100MHz,CDCl3):δ 165.51,139.33,134.86,133.55,133.17,132.58,130.40,129.51,129. 50,126.35,125.40,125.09,121.45,54.69,48.36,32.70,25.52,25.40. HRMS(ESI) m / z[M+H] + C 20 H 21 Calculated for BrN2O3S: 448.0539, found: 448.0267. LC-MS: Retention time 4.10 min.

[0216] Example 57 2-(8-Bromo-5,5-dioxide-6H-dibenzo[c,e][1,2]thiazin-6-yl)-N-cyclohexylacetamide (SM228) and 2-(10-Bromo-5,5-dioxide-6H-dibenzo[c,e][1,2]thiazin-6-yl)-N-cyclohexylacetamide (SM229): Following the general procedure described above, a mixture of the two regioisomers of formula 6a was reacted with cyclohexylamine to give the two regioisomers of formula 8a, which were separated by flash column chromatography (CHCl / acetone 98:2) and each compound was further purified by crystallization from EtOH to give the target compound SM228 (R by TLC). f >) and SM229 (R by TLC f <).

[0217] SM228: Yield 8%: Melting point 184-185°C. 1H-NMR (400MHz, CDCl3): δ 8.10-7.80(m,3H,Ar-H),7.82(t,J=7.4Hz,1H,Ar-H),7.65(t,J=7.6Hz,1H,Ar-H),7.52(dd,J=1.5 and 8.5 Hz, 1H,Ar-H),7.40(brs,1H,Ar-H),6.55(d,J=8.0Hz,1H,NH),4.50(s,2H,NCH2),4.00-3.75(m,1 H,Cy-CH),2.00-1.80(m,2H,Cy-CH2),1.75-1.50(m,2H,Cy-CH2),1.45-1.00(m,6H,Cy-CH2). 13 C NMR(100MHz,DMSO-d6):δ 165.60,140.02,134.99,133.22,131.50,129.45,128.16,127.94,126. 66,123.86,123.82,123.30,121.76,50.06,48.12,32.67,25.53,24.76. HRMS(ESI) m / z[M+Na] + C 20 H 21 Calculated for BrN2O3S: 471.0354, found: 471.041. LC-MS: Retention time 12.592 min.

[0218] SM229: Yield 21%: Melting point 211-212°C. 1 H-NMR(400MHz, CDCl3):δ 8.62(d,J=8.3Hz,1H,Ar-H),8.00(dd,J=1.4 and 7.8Hz,1H,Ar-H),7.75-7.65(m,4H,Ar-H),7.28-7.24(m,2H,Ar-H),6.50(d,J=8.1Hz,1 H,NH),4.40(s,2H,NCH2),3.80-3.70(m,1H,Cy-CH),1.90-1.80(m,2H,Cy-CH2),1.75-1.50(m,2H,Cy-CH2),1.45-1.00(m,6H,Cy-CH2). The NMR COSY spectrum showed two relevant NOE crosspeaks: H-9 (δ 7.70, dd) → H-8 (δ 7.32, t) and H-9 → H-7 (δ 7.28, dd). The NMR NOESY spectrum showed one relevant NOE crosspeak: H-8 → NCH2.13 C NMR(100MHz,DMSO-d6):δ 165.49,140.92,134.85,131.81,131.48,131.19,131.13,130.37,129. 41,125.44,121.64,121.38,120.59,50.77,48.11,32.66,25.52,24.76. HRMS(ESI) m / z[M+Na] + C 20 H 21 Calculated for BrN2O3S: 471.0354, found: 471.0407. LC-MS: Retention time 12.893 min.

[0219] Example 58 2-(8-chloro-5,5-dioxide-6H-dibenzo[c,e][1,2]thiazin-6-yl)-N-cyclohexylacetamide (SM586) and 2-(10-chloro-5,5-dioxide-6H-dibenzo[c,e][1,2]thiazin-6-yl)-N-cyclohexylacetamide (SM585): Following the general procedure described above, a mixture of the two appropriate regioisomers of formula 6a was reacted with cyclohexylamine to give the two regioisomers of formula 8a, which were separated by flash column chromatography (CHCl / acetone 98:2) and each compound was further purified by crystallization from EtOH to give the target compound SM586 (R by TLC). f >) and SM585 (R by TLC f <).

[0220] SM586: Yield 18%: Melting point 193-195°C. 1H-NMR (400MHz, CDCl3): δ8.02-7.90 (m, 3H, Ar-H), 7.74 (dt, J=1.2 and 7.7Hz, 1H, Ar-H), 7.60 ( dt,J=0.7 and 8.0Hz,1H,Ar-H),7.33(dd,J=2.0 and 8.2Hz,1H,Ar-H),7.28(d,J=2.0Hz,1H,Ar-H ),6.50(d,J=7.5Hz,1H,NH),4.49(s,2H,NCH2),3.90-3.75(m,1H,Cy-CH),1.90-1.80(m,2H, Cy-CH2),1.75-1.50(m,4H,Cy-CH2),1.45-1.35(m,2H,Cy-CH2),1.20-1.05(m,2H,Cy-CH2). 13 C-NMR (100MHz, CDCl3): δ 166.05,139.07,136.51,133.87,133.07,131.56,128.74,126.80,125. 71,125.68,122.52,122.41,119.71,51.57,48.41,32.41,25.36,24.30. HRMS(ESI) m / z[M+H] + C 20 H 21 Calculated value of ClN2O3S: 405.1039, measured value: 405.1037. LC-MS: Holding time 5.628 minutes.

[0221] SM585: Yield 15%: melting point 200~202℃. 1H-NMR (400MHz, CDCl3): δ 8.60(d,J=8.0Hz,1H,Ar-H),7.95(dd,J=1.2andび8.5Hz,1H,Ar-H),7.70(dt,J=1.3andび8.5Hz,1H,Ar-H),7.5 9(dd,J=1.2 and 8.0Hz,1H,Ar-H),7.43(dd,J=1.1 and 8.1Hz,1H,Ar-H),7.34(t,J=8.1Hz,1H,Ar-H),7.20(dd ,J=1.1andび8.1Hz,1H,Ar-H),6.35(d,J=7.0Hz,1H,NH),4.48(s,2H,NCH2),3.90-3.75(m,1H,Cy-CH),1.90 -1.80(m,2H,Cy-CH2),1.75-1.50(m,4H,Cy-CH2),1.45-1.35(m,2H,Cy-CH2),1.20-1.05(m,2H,Cy-CH2). 13 C-NMR (100MHz, CDCl3): δ 166.25,139.99,135.69,132.59,131.57,130.35,130.16,130.02,128. 78,128.70,123.48,122.29,118.69,52.15,48.38,32.45,25.34,24.33. HRMS(ESI) m / z[M+H] + C 20 H 21 Calculated value of ClN2O3S: 405.1039, measured value: 405.1037. LC-MS: Holding time 5.631 minutes.

[0222] Example 59 N-Cyclohexyl-2-[5,5-dioxido-8-(trifluoromethyl)-6H-dibenzo[c,e][1,2]thiazin-6-yl]acetamide (SM338) and N-cyclohexyl-2-[5,5-dioxido-10-(trifluoromethyl)-6H-dibenzo[c,e][1,2]thiazin-6-yl]acetamide (SM339): Following the general procedure described above, a mixture of the two appropriate regioisomers of formula 6a was reacted with cyclohexylamine to give the two regioisomers of formula 8a, which were separated by flash column chromatography (CHCl / acetone 99:1), and each compound was further purified by crystallization from EtOH to give the target compound SM338 (R by TLC). f >) and SM339 (R by TLC f <).

[0223] SM338: Yield 40%: Melting point 230-232°C. 1 H-NMR(400MHz, CDCl3):δ 8.20-8.10(d,J=8.2Hz,1H,Ar-H),8.05-7.98(m,2H,Ar-H),7.78(dt,J=1.5 and 7.5Hz,1H,Ar-H),7.7 8(dt,J=1.5 and 7.5Hz,1H,Ar-H),7.69(dt,J=1.3 and 7.7Hz,1H,Ar-H),7.65(d,J=8.0Hz,1H,Ar-H),7.5 0(brs,1H,Ar-H),6.50(d,J=6.9Hz,1H,NH),4.48(s,2H,NCH2),3.90-3.75(m,1H,Cy-CH),1.90-1.8 0(m,2H,Cy-CH2),1.75-1.50(m,4H,Cy-CH2),1.45-1.35(m,2H,Cy-CH2),1.20-1.05(m,2H,Cy-CH2). 13 C-NMR(100MHz,CDCl3):δ 165.86,138.57,134.66,133.16,132.50(q,J C-F =33.1Hz,C-8),131.11,129.58,127.08,126.42,126.27,123.79(q,J C-F =271.0Hz,CF3),122.59,121.89(q,J C-F=5Hz,C-9),116.83(q,J C-F =6Hz,C-7),51.71,48.44,32.40,25.33,24.33. HRMS(ESI) m / z[M+H] + C 21 H 21 Calculated for F₃N₂O₃S: 439.1303, found: 439.1296. LC-MS: retention time: 6.892 minutes.

[0224] SM339: Yield 26%: Melting point 211~212℃. 1 H-NMR (400MHz, CDCl3): δ 8.03-7.95(m,2H,Ar-H),7.82-7.58(m,4H,Ar-H),7.51(d,J=8.3Hz,1H,Ar-H),6.35(d,J=8.0Hz,1H,NH),4.40(s,2H ,NCH2),4.00-3.75(m,1H,Cy-CH),1.90-1.80(m,2H,Cy-CH2),1.75-1.50(m,4H,Cy-CH2),1.45-1.05(m,4H,Cy-CH2). 13 C-NMR (100MHz, CDCl3): δ 168.08,140.01(brs,C-6a),139.80,134.90,134.32,132.30,130.56,127.7(d,J C-F =2.0Hz,C-8),124.57,122.30(q,J C-F =29.0Hz,C-10),119.60(q,J C-F =270.1Hz,CF3),117.80(q,J C-F =3.1Hz,C-9),115.85(q,J C-F 2.0Hz,C-10a),115.09,52.02,49.50,32.40,26.28,24.12. HRMS(ESI) m / z[M+H] + C 21 H 21 Calculated for F₃N₂O₃S: 439.1303, found: 439.1298. LC-MS: retention time: 6.598 minutes.

[0225] Example 60 2-(8-chloro-9-fluoro-5,5-dioxide-6H-dibenzo[c,e][1,2]thiazin-6-yl)-N-cyclohexylacetamide (SM336) and 2-(10-chloro-9-fluoro-5,5-dioxide-6H-dibenzo[c,e][1,2]thiazin-6-yl)-N-cyclohexylacetamide (SM337): Following the general procedure described above, a mixture of the two appropriate regioisomers of formula 6a was reacted with cyclohexylamine to give the two regioisomers of formula 8a, which were separated by flash column chromatography (cyclohexane / EtOAc 7:3) and then crystallized from EtOH to give the target compound SM336 (R by TLC). f >) and SM337 (R by TLC f <).

[0226] SM336: Yield 21%: Melting point 211-213°C. 1 H-NMR(400MHz,DMSO-d6):δ 7.96(dd,J=1.5 and 7.0Hz,1H,Ar-H),7.8(d,J=8.2Hz,1H,Ar-H),7.78-7.70(m ,2H,Ar-H),7.61(dt,J=1.4 and 7.7Hz,1H,Ar-H),7.33(d,J=6.4Hz,1H,Ar-H),6 .4(d,J=9.0Hz,1H,Ar-H),4.35(s,2H,NCH2),3.80-3.70(m,1H,Cy-CH),1.85 -1.75(m,2H,Cy-CH2),1.75-1.45(m,2H,Cy-CH2),1.45-1.00(m,6H,Cy-CH2). 13 C NMR(100MHz,DMSO-d6):δ 165.59,154.86(d,J C-F =243.0Hz,C-9), 136.05(d,J C-F =2.6Hz,C-6a),135.17,133.19,130.80,130.05,127.16,125.69(d,J C-F =8.0Hz,C-10a),123.97,121.84,121.29(d,J C-F =20.0Hz,C-8), 113.96(d,J C-F= 24.0 Hz, C-10), 50.97, 48.12, 32.62, 25.51, 24.73. HRMS(ESI) m / z [M+H] + C 20 H 20 Calculated value of ClFN2O3S: 423.0946, measured value: 423.0938. LC-MS: retention time 6.560 min.

[0227] SM337: Yield 53%: melting point 216 - 217 °C. 1 1H-NMR(400 MHz, CDCl3): δ 8.51 (d, J = 8.2 Hz, 1H, Ar-H), 8.00 (d, J = 7.7 Hz, 1H, Ar-H), 7.73 (dt, J = 1.2 and 7.5 Hz, 1H, Ar-H), 7.64 (t, J = 7.5 Hz, 1H, Ar-H), 7.35 - 7.20 (m, 2H, Ar-H), 6.25 (d, J = 7.1 Hz, 1H, NH), 4.25 (s, 2H, NCH2), 3.80 - 3.70 (m, 1H, Cy-CH), 1.90 - 1.80 (m, 2H, Cy-CH2), 1.75 - 1.50 (m, 4H, Cy-CH2), 1.40 - 1.00 (m, 4H, Cy-CH2). 13 13C NMR(100 MHz, CDCl3): δ 166.08, 156.77 (d, J C-F = 246.1 Hz, C-9), 135.85 (brs, C-6a), 135.81, 131.72, 129.99, 129.81 (d, J C-F = 2.8 Hz, C-10a), 129.39, 125.44, 122.69, 120.41 (J C-F = 8.1 Hz, C-7), 119.5 (J C-F = 20.1 Hz, C-10), 117.43 (J C-F = 24.0 Hz, C-8), 52.73, 48.45, 32.53, 25.33, 24.39. HRMS(ESI) m / z [M+H] + C 20 H 20 Calculated value of ClFN2O3S: 423.0946, measured value: 423.0938. LC-MS: retention time 6.520 min.

[0228] Example 61 N-Cyclohexyl-2-(8,10-dichloro-5,5-dioxide-6H-dibenzo[c,e][1,2]thiazin-6-yl)acetamide (SM587): Following the general procedure described above, the title compound was obtained after crystallization from EtOH in 50% yield as a white solid, mp 212.0-213.0 °C. 1 H-NMR(200MHz,DMSO-d6)δ 8.50(d,J=8.0Hz,1H,Ar-H),7.97(dd,J=1.3 and 7. 8H z, 1H, Ar-H), 7.71 (dt, J = 1.4 and 8.0 Hz, 1H, Ar-H), 7.61 (dt, J = 0.9 and 7. 6H z,1H,Ar-H),7.45(d,J=2.0Hz,1H,Ar-H),7.22(d,J=2.0Hz,1H,Ar-H),6.28(d,J=7.6Hz,1H,NH),4.31(s,2H,NCH 2),3.80-3.70(m,1H,Cy-CH),1.90-1.80(m,2H,Cy-CH2),1.75-1.50(m,4H,Cy-CH2),1.40-1.00(m,4H,Cy-CH2). 13 C NMR(100MHz,CDCl3):δ 165.74,140.50,135.53,135.44,133.58,131.78,129.81,129.75,129. 03,128.46,122.36,122.06,119.02,52.00,48.47,32.46,25.32,24.37. HRMS(ESI) m / z[M+H] + C 20 H 20 Calculated for Cl2N2O3S: 439.0649, found: 439.0646. LC-MS: Retention time 1.920 min.

[0229] Example 62 2-(5,5-Dioxido-6H-dibenzo[c,e][1,2]thiazin-6-yl)-N-cyclohexylacetamide (SM7): To a suspension of LiAlH (0.021 g, 0.55 mmol) in dry THF (1 mL) cooled to 0 °C, a solution of SM3 (0.100 g, 0.22 mmol) in dry THF (4 mL) was added dropwise under N, and the mixture was stirred at 50 °C for 2 h. After cooling and quenching with EtOAc followed by MeOH, the mixture was poured into ice water and extracted with EtOAc (3 × 20 mL). The combined organic layers were washed with brine, dried, and evaporated to dryness. The resulting crude colorless oil was purified by flash column chromatography eluting with cyclohexane / EtOAc (7:3) to give SM7 (0.040 g, 49%) as a white solid. mp 176–178 °C. 1 H NMR (400 MHz, CDCl3): δ 8.25-8.00 (m, 3H, Ar-H), 7.77 (dt, J = 1.2 and 8.4 Hz, 1H, Ar-H), 7.62 (t, J = 7.7 Hz, 1H, Ar-H), 7.51 (dt, J = 1.3 and 8.6 Hz, 1H, Ar-H), 7.39 (dt, J = 1.0 and 8.4 Hz, 1H, Ar-H), 7.25 (dd, J = 1.8 and 7.2 Hz ,1H,Ar-H),6.60(brs,1H,NH),4.48(s,2H,N-CH2),3.91-3.83(m,1H,Cy-CH),1.90-1.80(m, 2H,Cy-CH2),1.60-1.50(m,3H,Cy-CH),1.40-1.25(m,2H,Cy-CH2),1.20-1.00(m,3H,Cy-CH). 13 C NMR(100MHz,CDCl3):δ 166.18,137.66,133.57,132.43,131.81,130.31,127.98,125.31,125. 21,124.88,123.36,121.96,118.99,51.19,47.85,31.92,24.87,23.81. HRMS(ESI) C 20 H 22 N2O3S [M + +H] + Calculated for: 371.1429, Found: 371.1397. LC-MS: Retention time 5.212.

[0230] Example 63 N-Cyclohexyl-2-(9-hydroxy-5,5-dioxide-6H-dibenzo[c,e][1,2]thiazin-6-yl)acetamide (SM226): To a solution of the target compound SM225 (0.22 g, 0.55 mmol) in dry CHCl (12 mL) under N flux, 1 M BBr in CHCl (2.75 g, 2.75 mmol) was added dropwise at −60° C., and the solution was then stirred at −30° C. for 12 h. After quenching the excess BBr with MeOH, HO, and saturated NaHCO solution, the mixture was adjusted to pH 8. 2 The resulting mixture was acidified with 2 N HCl to rt and extracted with CHCl (3 × 30 mL). The combined organic layers were washed with brine, dried, and evaporated to dryness, and the residue was purified by flash column chromatography (CHCl / MeOH 95:5) to give compound SM226 in 88% yield as a white solid, mp 216–217 °C. 1 H-NMR(400MHz,DMSO-d6):δ 9.77(s,1H,OH),8.15(d,J=8.9Hz,1H,Ar-H),7.90-7.74(m,3H,Ar-H and NH),7.62(t,J=7.6Hz,1H,Ar-H),7.45(d,J=2.5Hz,1H,Ar-H),7.25(d,J=8.8H) z,1H,Ar-H),6.85(dd,J=2.6 and 8.7Hz,1H,Ar-H),4.30(s,2H,NCH2),3.40- 3.30(m,1H,Cy-CH),1.75-1.40(m,5H,Cy-CH2),1.30-0.90(m,5H,Cy-CH2). 13 C-NMR(100MHz,DMSO-d6):δ 166.19,154.95,143.08,135.01,133.28,132.00,129.29,126.41,126. 31,123.94,121.99,118.10,111.45,51.74,48.13,32.38,25.31,24.59. HRMS(ESI) m / z[M+H] + C 20 H 22 Calculated for N2O4S: 387.1380, found: 387.1372. LC-MS: Retention time 4.691 min.

[0231] Example 64 N-Cyclohexyl-2-[9-[2-(dimethylamino)ethoxy]-5,5-dioxide-6H-dibenzo[c,e][1,2]thiazin-6-yl]acetamide (SM230): To a solution of the target compound SM226 (0.18 g, 0.47 mmol) in dry DMF (7 mL) was added CsCO (0.23 g, 0.70 mmol) and commercial 1-chloro-N,N-dimethylethanamine hydrochloride (0.07 g, 0.47 mmol). The mixture was maintained at 85 °C for 2 h under magnetic stirring. The mixture was poured into ice water and extracted with EtOAc (3 × 20 mL). The combined organic layers were washed with brine, dried, and evaporated to dryness to give an oil, which was purified by flash column chromatography (CHCl / MeOH 9:1) to give SM230 in 57% yield as a low-melting solid: mp 66–67 °C. 1 H-NMR (200 MHz, DMSO-d): δ 8.23(d,J=8.1Hz,1H,Ar-H),7.90-7.65(m,3H,Ar-H and NH),7.60-7.55(m,2H,Ar-H) ,7.28(d,J=8.9Hz,1H,Ar-H),7.10(dd,J=2.5 and 9.0Hz,1H,Ar-H),4.40(s,2H,SO2N -CH2),4.20(t,J=5.3Hz,2H,OCH2),3.90-3.80(m,1H,Cy-CH),2.80(t,J=5.3Hz,2H ,NCH2),2.40(s,6H,NCH3),1.75-1.40(m,5H,Cy-CH2),1.30-0.90(m,5H,Cy-CH2). 13 C-NMR(100MHz,DMSO-d6):δ 165.84,156.34,135.42,132.86,132.29,132.26,129.15,126.89,126.26,123 .65,121.87,117.63,110.75,66.68,51.40,48.04,45.99,32.66,25.53,24.77. HRMS(ESI) m / z[M+H] + C 24 H 31 Calculated for N3O4S: 458.2200, found: 458.2200. LC-MS: Retention time 6.360 min.

[0232] Below, the experimental procedure for making compounds of formula 7a in Scheme 1 is described.

[0233] Example 65 2-(9-Bromo-5,5-dioxide-6H-dibenzo[c,e][1,2]thiazin-6-yl)acetic acid of Formula 7a: A mixture of 2-(9-bromo-5,5-dioxide-6H-dibenzo[c,e][1,2]thiazin-6-yl)ethyl acetate of Formula 6a (Example 36; 0.600 g, 1.5 mmol) in 10% aqueous NaOH (7 mL) and EtOH (7 mL) was refluxed for 30 min, then cooled, concentrated under reduced pressure, poured into ice water, and acidified with 2N HCl to pH 2. The precipitate that formed was filtered to give the compound (0.540 g, 96%) as a white solid that was used directly in the next reaction step; melting point 207-209 °C. 1 H NMR(400MHz,DMSO-d6):δ 8.40(d,J=2.2Hz,1H,Ar-H),8.31(d,J=8.0Hz,1H,Ar-H),7.91(dd,J=1.1 and 7.7Hz,1H,Ar-H),7.84(d t,J=1.3 and 7.7Hz,1H,Ar-H),7.75-7.69(m,2H,Ar-H),7.45(d,J=8.7Hz,1H,H-7),4.80(s,2H,NCH2).

[0234] Example 66 3-Fluoro-5,5-dioxide-9-(trifluoromethyl)-6H-dibenzo[c,e][1,2]thiazin-6-yl]acetic acid of formula 7a: To a solution of ethyl [3-fluoro-5,5-dioxide-9-(trifluoromethyl)-6H-dibenzo[c,e][1,2]thiazin-6-yl]acetate of formula 6a (Example 43; 1.25 g, 3.09 mmol) in dioxane (25 mL) was added a solution of 1N LiOH monohydrate (2.47 mL). The reaction mixture was stirred at room temperature for 10 minutes, then poured into ice water and acidified (pH = 2) with 2N HCl. The precipitate formed was filtered and dried to give the desired compound as a white solid (1.16 g, 96%). 1H NMR(400MHz,CDCl3):δ 4.70(s,1H,NCH2),7.35(d,J=8.5Hz,1H,H-7),7.40-7.50(m,1H,H-2),7.60-7.65(m,1H ,H-4),7.70(d,J=8.5Hz,1H,H-8),8.00(dd,J=4.5 and 8.8Hz,1H,H-1),8.20(s,1H,H-10).

[0235] Example 67 2-(9-Bromo-5,5-dioxo-6H-dibenzo[c,e][1,2]thiazin-6(5H)-yl)-N-phenylacetamide (SM6). A mixture of 2-(9-bromo-5,5-dioxo-6H-dibenzo[c,e][1,2]thiazin-6-yl)acetic acid (Example 65) (0.530 g, 1.44 mmol) of general formula 7a and SOCl (2 mL) was refluxed under magnetic stirring for 1 h, and then excess SOCl was removed by distillation, and the residue was washed three times with dry toluene. The resulting acyl chloride was solubilized in dry DMF (7 mL) and added dropwise to a stirred solution of aniline (0.264 mL, 2.88 mmol) and EtN (0.401 mL, 2.88 mmol) in dry DMF (3 mL) at room temperature under a N atmosphere. The mixture was left under magnetic stirring overnight, then poured into ice water and acidified with 2 N HCl to pH 3. The precipitate was filtered and purified by flash column chromatography eluting with CHCl to give the target compound SM6 (0.150 g, 25%) as a white solid: melting point 128-130 °C. 1 H NMR (400 MHz, CDCl): δ 8.37(bs,1H,NH),),8.20(d,J=2.1Hz,1H,Ar-H),8.10(d,J=7.7Hz,1H,Ar-H),8.0 0(d,J=7.7Hz,1H,Ar-H),7.80(t,J=7.7Hz,1H,Ar-H),7.70(t,J=7.7Hz,1H,Ar-H) ,7.60(dd,J=2.2 and 7.8Hz,1H,Ar-H),7.55-7.48(m,2H,Ar-H),7.35-7.30(m,2H,A r-H),7.20(d,J=7.8Hz,1H,Ar-H),7.10(t,J=7.4Hz,1H,Ar-H),4.52(s,2H,CH2). 13C NMR(100MHz,CDCl3):δ 165.47,137.03,136.86,133.88,133.75,133.33,131.09,129.30,129.06 ,128.73,126.02,125.66,125.09,122.76,121.21,120.04,118.98,52.02. HRMS(ESI) m / z[M+H] + C 20 H 15 Calculated for BrN2O3S: 443.0069, found: 443.0057. LC-MS: retention time 5.571.

[0236] Example 68 2-(9-Bromo-5,5-dioxide-6H-dibenzo[c,e][1,2]thiazin-6-yl)-N-cyclohexyl-N-methylacetamide. The appropriate compound of general formula 7a (2-(9-Bromo-5,5-dioxide-6H-dibenzo[c,e][1,2]thiazin-6-yl)acetic acid; Example 65) (0.59 g, 1.6 mmol) was chlorinated as described above, and the corresponding acyl chloride was dissolved in dry DMF (8 mL) and added dropwise to a solution of N-methylcyclohexylamine (0.83 mL, 6.4 mmol) in dry DMF (2 mL) at room temperature under a N atmosphere. The mixture was heated at 40° C. for 1.5 hours, then poured into ice water and acidified to pH 3 with 2 N HCl. The precipitate was filtered and purified by flash column chromatography eluting with CHCl 3 , followed by trituration with petroleum ether / Et 2 O to give the target compound SM8 (0.197 g, 30%) as a white solid: mp 170-172° C. 1H NMR (400MHz, DMSO-d6): (mixture of rotamers) δ 8.42(d,J=1.7Hz,1H,Ar-H),8.30(d,J=8.0Hz,1H,Ar-H),7.87(d,J=7.8Hz,1H,Ar-H), 7.82(t,J=7.6Hz,1H,Ar-H),7.75-7.65(m,2H,Ar-H),7.43(t,J=8.9Hz,1H,H-7),4.97( s,0.88H,NCH2),4.90(s,1.12H,NCH2),4.00-3.90(m,0.54H,Cy-CH),3.60-3.50(m,0. 46H,Cy-CH),2.80(s,1.68H,NCH3),2.60(s,1.32H,NCH3),1.75-0.95(m,10H,Cy-CH2). 13 C NMR (100MHz, DMSO-d6): (mixture of rotamers) δ 165.91,165.87,138.45,138.32,135.56,135.52,133.15,133.11,132.94,132.91,131.09,129.59,128.36,126.96,126.92,1 23.76,123.73,121.54,121.48,117.71,117.65,55.11,52.84,50.10,30.59,29.48,28.68,27.38,25.61,25.45,25.30,25.15. HRMS(ESI) C 21 H 23 BrN2O3S [M + +H] + Calculated for: 463.0692, found: 463.0678. LC-MS: retention time 6.034. After recording the NMR spectrum at 50°C, the two rotamers had collapsed into one molecule.

[0237] Example 69

[0238] [ka]

[0239] 2-(3-Fluoro-5,5-dioxide-9-(trifluoromethyl)-6H-dibenzo[c,e][1,2]thiazin-6-yl)acetic acid (7a(Int-1)) and 2-(3-ethoxy-5,5-dioxide-9-(trifluoromethyl)-6H-dibenzo[c,e][1,2]thiazin-6-yl)acetic acid (7a(Int-2)): A mixture of the compound of formula 6a(Int-1) (0.40 g, 0.99 mmol) in 10% aqueous NaOH (3 mL) and EtOH (3 mL) was refluxed for 30 min. After cooling, the organic solvent was evaporated under reduced pressure, and the residue was poured into ice water and acidified (pH = 2) with 2 N HCl. The precipitate formed was filtered to give a mixture of two compounds 7a (Int-1) and 7a (Int-2) in a 1:1 ratio, as highlighted by the presence of two spots in TLC (CHCl3:MeOH 8:2), 1 This was also confirmed by H-NMR spectroscopy. 1 H NMR(400MHz,CDCl3):δ 8.20(bs,0.5H,H-10),8.17(bs,0.5H,H-10),7.90(dd,J=5 and 9Hz,0.5H,H-1),7.85(d,J=9 Hz,0.5H,H-1),7.73-7.60(m,1H,H-4 and H-8),7.60(d,J=8.5Hz,0.5H,H-8),7.45(m,0.5H,H -2), 7.40 (s, 0.5H, H-4), 7.35 (d, J = 8 Hz, 0.5H, H-7), 7.20-7.30 (m, 1H, H-2 and H-7), 4.67 (s, 1H, N-CH2), 4.65 (s, 1H, N-CH2), 4.10 (q, J = 7.0 Hz, 1H, OCH2), 1.45 (t, J = 7.0 Hz, 1.5H, CH3). Compounds 7a (Int-1) and 7a (Int-2) were obtained as orange solids and used directly in the subsequent amidation step.

[0240] Example 70 Scheme 6: Preparation of target compounds derived from intermediates of general formula 7a not included in scheme 1

[0241] [ka]

[0242] N-Cyclohexyl-2-(3-fluoro-5,5-dioxide-9-(trifluoromethyl)-6H-dibenzo[c,e][1,2]thiazin-6-yl)acetamide (SM882) and N-cyclohexyl-2-(3-ethoxy-5,5-dioxide-9-(trifluoromethyl)-6H-dibenzo[c,e][1,2]thiazin-6-yl)acetamide (SM883): A stirred mixture of 7a (Int-1) and 7a (Int-2) (0.50 g, 1.33 mmol), cyclohexylamine (0.18 mL, 1.6 mmol), TBTU (0.55 g, 1.7 mmol), and DIPEA (0.93 mL, 5.33 mmol) in dry CHCl (3 mL) was reacted at room temperature for 1 h. The solvent was then evaporated to dryness, the residue was poured into ice water, the resulting precipitate was filtered, and the crude was purified by flash chromatography eluting with CH2Cl2 to give SM882(R f >) and SM883(R f Each compound was purified by crystallization with EtOH to give the following:

[0243] SM882: White solid (0.064 g, 14%), melting point 232-233°C. 1 H NMR (400 MHz, CDCl3): δ 1.10-1.20 and 1.30-1.40 (m, each 2H, cyclohexyl CH2), 1.50-1.70 (m, 4H, cyclohexyl CH2), 1.80-1.90 (m, 2H, cyclohexyl CH2), 3.85-3.95 (m, 1H, cyclohexyl CH), 4.55 (s, 1H, N CH2),6.45(d,J=7.5Hz,1H,CONH),7.40(d,J=8.5Hz,1H,Ar-H),7.55(dt,J=2.6 and 8.1Hz,1 H, Ar-H), 7.75-7.85 (m, 2H, Ar-H), 8.10 (dd, J=4.6 and 8.9Hz, 1H, Ar-H), 8.30 (s, 1H, Ar-H). 13 C NMR(101MHz,CDCl3):δ 24.2,25.2,32.4,48.5,51.4,109.8(d,J C-F =25.4Hz), 119.8, 120.8(d,JC-F =22.2Hz),122.7(d,J C-F =3.6Hz),123.5,128.8(q,J C-F =273.3Hz),127.1(d,J C-F =3.3Hz),127.4,127.7,128.5(d,J C-F =8.1Hz),135.4(d,J C-F =7.3Hz),140.1,162.2(d,J C-F =256.9Hz),165.6. HRMS(ESI) m / z[M+H] + C 21 H 20 Calculated value of F4N2O3S: 457.1210, measured value: 457.1207.

[0244] SM883: white solid (0.069g, 15%), melting point 201~202℃. 1 H NMR (400MHz, CDCl3): δ1.10-1.20(m,4H,シクロヘキシルCH2),1.30-1.40(m,2H,シクロヘキシルCH2),1.50(t,J=6.9 Hz, 3H, OCH2CH3), 1.60-1.70 and 1.80-1.90 (m, each 2H, シクロヘキシルCH2), 3.80-3.90 (m, 1H, シクロヘキシルCH), 4.20( q,J=6.9Hz,2H,OCH2CH3),4.55(s,1H,NCH2),6.55(d,J=7.7Hz,1H,CONH),7.30-7.40(m,2H,Ar-H),7. 50(d,J=2.1Hz,1H,Ar-H),7.70(d,J=8.6Hz,1H,Ar-H),7.95(d,J=8.9Hz,1H,Ar-H),8.25(s,1H,Ar-H). 13 C NMR (101MHz, CDCl3): δ 14.4, 24.2, 25.2, 32.3, 48.4, 51.2, 64.4, 106.1, 119.3, 121.1, 122.1 (d, J C-F =3.6Hz),123.9,126.1(d,J C-F =3.3Hz),127.5,134.9,139.6,159.6,166.0. HRMS(ESI) m / z[M+H] + C 23 H 25Calculated value of F3N2O4S: 483.1566, measured value: 483.1565.

[0245] Example 71 N-(1-Ethylpropyl)-2-[3-fluoro-5,5-dioxide-9-(trifluoromethyl)-6H-dibenzo[c,e][1,2]thiazin-6-yl]acetamide (SM884): A stirred mixture of 3-fluoro-5,5-dioxide-9-(trifluoromethyl)-6H-dibenzo[c,e][1,2]thiazin-6-yl]acetic acid of Formula 7a (Example 63; 0.30 g, 0.8 mmol), 3-aminopentane (0.084 g, 0.96 mmol), TBTU (0.33 g, 1.04 mmol), and DIPEA (0.56 mL, 3.2 mmol) in CHCl (6 mL) was kept at room temperature for 3 hours. The organic solvent was evaporated, the residue was poured into ice / water, and the mixture was acidified (pH = 4) with 2 N HCl. The mixture was kept under stirring for 40 minutes until a precipitate was observed. The precipitate was filtered, dried and crystallized from cyclohexane / EtOAc (3:1 ratio) to give SM884 in 34% as a pinkish solid: mp 184-185°C. 1 H NMR (400 MHz, CDCl): δ 0.80 (t, J = 7.4 Hz, 6H, pentyl CH), 1.30-1.40 and 1.45-1.55 (m, each 2H, pentyl CH), 3.75-3.80 (m, 1H, pentyl CH), 4.50 (s, 1H, NCH), 6.25 (d, J = 8.6 Hz, 1H, CONH), 7.40 (d, J = 8.6 Hz, 1H, Ar-H), 7.50 (dt, J = 2.6 and 8.3 Hz, 1H, Ar-H), 7.70-7.75 (m, 2H, Ar-H), 8.10 (dd, J = 4.6 and 8.9 Hz, 1H, Ar-H), 8.25 (s, 1H, Ar-H). 13 C NMR(100MHz,CDCl3):δ 9.9,27.0,51.4,52.7,109.8(d,J C-F =25.5Hz), 119.8, 120.9(d,J C-F =22.3Hz), 122.7(d,J C-F =3.5Hz), 123.4, 126.1(q, J C-F=273.0Hz), 127.1(d,J C-F =3.2Hz), 127.7, 128.6(d,J C-F =8.1Hz), 135.3(d,J C-F =7.3Hz), 140.1, 162.2(d,J C-F =257.0Hz),166.4. HRMS(ESI) m / z[M+H] + C 20 H 20 Calculated value of F4N2O3S: 445.1210, measured value: 445.1207.

[0246] Example 72 2-[3-Fluoro-5,5-dioxide-9-(trifluoromethyl)-6H-dibenzo[c,e][1,2]thiazin-6-yl]-N-(tetrahydro-2H-pyran-4-yl)acetamide (SM885): Following the procedure set out above for compound SM884, tetrahydro-2H-pyran-4-amine was used to give the target compound after crystallization from cyclohexane / EtOAc in 34% yield as a pale pink solid: mp 241-242 °C. 1 H NMR (400 MHz, CDCl): δ 1.40-1.50, 1.80-1.90, 3.40-3.50, and 3.75-3.85 (m, each 2H, pyran CH), 4.00-4.10 (m, 1H, pyran CH), 4.50 (s, 1H, NCH), 6.45 (d, J = 7.4 Hz, 1H, CONH), 7.40 (d, J = 8.6 Hz, 1H, Ar-H), 7.50 (dt, J = 2.7 and 8.5 Hz, 1H, Ar-H), 7.65-7.75 (m, 2H, Ar-H), 8.05 (dd, J = 4.6 and 8.9 Hz, 1H, Ar-H), 8.25 (s, 1H, Ar-H). 13 C NMR(100MHz,CDCl3):δ 32.4,46.0,51.3,66.2,109.8(d,J C-F =25.5Hz), 119.7, 121.0(d,J C-F =22.2Hz), 122.7(d,J C-F =3.5Hz), 123.4, 126.1(q, J C-F =273.0Hz), 127.2, 127.4(d,JC-F =3.2Hz), 127.8, 128.6(d,J C-F =8.1Hz), 135.3(d,J C-F =7.3Hz), 140.0, 162.2(d,J C-F =257.2Hz),166.0. HRMS(ESI) m / z[M+H] + C 20 H 18 Calculated value of F4N2O4S: 459.1002, Measured value: 459.1002.

[0247] Example 73 2-[3-Fluoro-5,5-dioxide-9-(trifluoromethyl)-6H-dibenzo[c,e][1,2]thiazin-6-yl]-N-morpholin-4-ylacetamide (SM881): Following the procedure given for compound SM884, morpholin-4-amine was used to give the target compound in 34% yield after crystallization from EtOH as a pale pink solid: melting point 276-278 °C. 1 Two rotamers were identified by H-NMR, and when the experiment was carried out at 60 °C, these collapsed into a single molecule. 1 H NMR (400 MHz, DMSO-d6, 25 °C): δ 2.50-2.60, 2.75-2.95, 3.40-3.50, and 3.60-3.80 (m, each 2H, morpholine CH2), 4.50 and 5.00 (s, each 1H, NCH2), 7.60-7.75 (m, 2H, Ar-H), 7.75-7.80 and 7.80-7.90 (m, each 1H, Ar-H), 8.45 (dd, J = 4.6 and 8.6 Hz, 1H, Ar-H), 8.55 (s, 1H, Ar-H), 8.80 and 9.25 (s, each 0.5H, CONH). 13 C NMR(100MHz,DMSO-d6):δ 32.4,46.0,66.2,110.1(d,J C-F =25.5Hz), 119.7, 122.0(d,J C-F =22.2Hz), 123.7(d,J C-F =3.5Hz), 123.4, 125.1(q, J C-F =273.0Hz), 127.2, 127.4(d,J C-F=3.2Hz), 127.8, 129.2(d,J C-F =8.1Hz), 134.2(d,J C-F =7.3Hz), 140.0, 161.2(d,J C-F =257.2Hz),166.0. HRMS(ESI) m / z[M+H] + C 19 H 17 Calculated value of F4N3O4S: 460.0955, measured value: 460.0954.

[0248] Example 74 N-(2-chloropyridin-4-yl)-2-[5,5-dioxide-9-(trifluoromethyl)-6H-dibenzo[c,e][1,2]thiazin-6-yl]acetamide (SM880): Starting from 2-(5,5-dioxide-9-(trifluoromethyl)-6H-dibenzo[c,e][1,2]thiazin-6-yl)acetic acid of formula 7a (Example 75), the title compound was prepared using 2-chloro-4-pyridinamine according to the procedure described for compound SM884. After crystallization from cyclohexane / EtOAc, the title compound was obtained in 34% yield as a pale pink solid: melting point 184-185 °C. 1 H NMR (400 MHz, CDCl): δ 0.80 (t, J = 7.4 Hz, 6H, pentyl CH), 1.30-1.40 and 1.45-1.55 (m, each 2H, pentyl CH), 3.75-3.80 (m, 1H, pentyl CH), 4.50 (s, 1H, NCH), 6.25 (d, J = 8.6 Hz, 1H, CONH), 7.40 (d, J = 8.6 Hz, 1H, Ar-H), 7.50 (dt, J = 2.6 and 8.3 Hz, 1H, Ar-H), 7.70-7.75 (m, 2H, Ar-H), 8.10 (dd, J = 4.6 and 8.9 Hz, 1H, Ar-H), 8.25 (s, 1H, Ar-H). 13 C NMR(101MHz,CDCl3):δ 9.9,27.0,51.4,52.7,109.8(d,J C-F =25.5Hz), 119.8, 120.9(d,J C-F =22.3Hz), 122.7(d,J C-F =3.5Hz), 123.4, 126.1(q, J C-F=273.0Hz), 127.1(d,J C-F =3.2Hz), 127.7, 128.6(d,J C-F =8.1Hz), 135.3(d,J C-F =7.3Hz), 140.1, 162.2(d,J C-F =257.0Hz),166.4. HRMS(ESI) m / z[M+H] + C 20 H 20 Calculated value of F4N2O3S: 445.1210, measured value: 445.1207.

[0249] Scheme 7: Preparation of target compounds derived from intermediates of general formula 7a not included in Scheme 1

[0250] [ka]

[0251] Example 75 2-(5,5-Dioxido-9-(trifluoromethyl)-6H-dibenzo[c,e][1,2]thiazin-6-yl)acetic acid (7a(Int-3)): The compound of formula 7a(Int-3) was prepared from ethyl [5,5-dioxido-9-(trifluoromethyl)-6H-dibenzo[c,e][1,2]thiazin-6-yl]acetate according to the procedure described for the similar compound in Example 65. The intermediate was obtained as a brown solid in 81% yield. 1 H NMR (400MHz, DMSO-d6): δ 8.55(d,J=2.2Hz,1H,Ar-H),8.27(d,J=8.0Hz,1H,Ar-H),8.00-7.75(m,4H,Ar-H),7.50-7.50(m,2H,Ar-H),4.75(s,2H,NCH2).

[0252] Example 76 N-(1-benzylpiperidin-4-yl)-2-[5,5-dioxide-9-(trifluoromethyl)-6H-dibenzo[c,e][1,2]thiazin-6-yl]acetamide of Formula 8a (Int-3): To a solution of 2-(5,5-dioxide-9-(trifluoromethyl)-6H-dibenzo[c,e][1,2]thiazin-6-yl)acetic acid of Formula 7a (Int-3) (Example 75; 0.280 g, 0.78 mmol) in dry CHCl (10 mL) was added N-benzyl-4-aminopiperidine (0.180 g, 0.94 mmol), TBTU (0.376 g, 0.12 mmol), and DIPEA (0.510 mL, 0.31 mmol). The reaction mixture was stirred at room temperature for 4 h, then poured into ice water and acidified (pH = 2) with 2 N HCl. The mixture was extracted with CHCl (3 × 30 mL), and the combined organic layers were washed with brine, dried over NaSO, and evaporated to dryness to give a brown oil, which was purified by flash column chromatography (CHCl / MeOH 95:5) to give N-(1-benzylpiperidin-4-yl)-2-[5,5-dioxide-9-(trifluoromethyl)-6H-dibenzo[c,e][1,2]thiazin-6-yl]acetamide as a solid in 45% yield: melting point 216~217 °C. 1 H-NMR (200 MHz, CDCl3): δ 8.27 (s, 1H, Ar-H), 8.03-7.99 (d, J = 7.9 Hz, 2H, Ar-H), 7.82-7.60 (m, 3H, Ar-H), 7.32-7.15 (m, 6H, Ar-H), 6.53 (d, J = 7.3 Hz, 1H, NH), 4.54 (s, 2H, benzyl-CH2), 4.83-4.71 (m, 1H, piperidine-CH), 3.45 (s, 2H, CH2), 2.67-2.62 (m, 2H, piperidine-CH2), 2.15-1.80 (m, 6H, piperidine-CH2 × 2), 1.47-1.32 (m, 2H, piperidine-CH2).

[0253] Example 77 2-[5,5-Dioxido-9-(trifluoromethyl)-6H-dibenzo[c,e][1,2]thiazin-6-yl]-N-piperidin-4-ylacetamide (SM655) of general formula 8a: To a solution of the appropriate compound of formula 8a (0.180 g, 0.34 mmol) in EtOH (20 mL) was added Pd / C (20% w / w, 0.036 g). The reaction mixture was stirred at room temperature under H bubbling for 7 hours. The mixture was filtered through Celite®, and the filtrate was evaporated to dryness to give a brown solid, which was crystallized from EtOH to give SM6 5 5 was obtained in 27% yield as a white solid. 1 H NMR (400 MHz, DMSO-d): δ 8.55 (s, 1H, Ar-H), 8.38-8.35 (m, 2H, Ar-H), 7.91-7.81 (m, 3H, Ar-H), 7.71 (t, J = 7.6 Hz, 1H, Ar-H), 7.62 (d, J = 8.6 Hz, 1H, Ar-H), 4.63 (s, 2H, benzyl-CH), 3.70-3.59 (m, 1H, piperidine-CH), 3.16-3.13 (m, 2H, piperidine-CH), 2.82 (t, J = 10.7 Hz, 2H, piperidine-CH), 1.77-1.74 (m, 2H, piperidine-CH), 1.49-1.37 (m, 2H, piperidine-CH).

[0254] Scheme 8: Preparation of target compounds derived from intermediates of general formula 7a not included in scheme 1

[0255] [ka]

[0256] Example 78 2-[5,5-Dioxido-9-(trifluoromethyl)-6H-dibenzo[c,e][1,2]thiazin-6-yl]-N-(trans-4-hydroxycyclohexyl)acetamide of Formula 8a (Int-4): To a solution of the compound of Formula 7a (Int-3) (Example 75; 0.100 g, 0.30 mmol) in dry CHCl (4 mL) was added trans-4-aminocyclohexanol (0.041 g, 0.36 mmol), BOP (0.199 g, 0.45 mmol), and DIPEA (0.200 mL, 1.2 mmol) at 0° C. The reaction mixture was stirred at room temperature for 12 hours, then concentrated in vacuo, poured into ice water, and acidified (pH=4) with 2 N HCl. The mixture was extracted with EtOAc (3 × 20 mL), and the combined organic layers were washed with brine, dried over NaSO, and evaporated to dryness to give a brown solid, which was crystallized from EtOH to give SM588 8a (Int-4) in 88% yield as a white solid: mp 212–213 °C. 1 H-NMR(400MHz, CDCl3):δ 8.29(d,J=1.3Hz,1H,Ar-H),8.03(d,J=7.3Hz,2H,Ar-H),7.81(td,J=1.3 and 7.4Hz,1H,Ar-H),7.72(dd,J= 1.6 and 6.3Hz,1H,Ar-H),6.67(t,J=7.5Hz,1H,Ar-H),7.33(d,J=7.9Hz,1H,Ar-H),6.52(d,J=7.6Hz,1H,NH ), 4.51 (s, 2H, benzyl-CH2), 3.85-3.77 (m, 1H, cyclohexyl-CH2), 3.61-3.46 (m, 1H, cyclohexyl-CH2), 1.99-1.89 (m, 4H, cyclohexyl-CH2 × 2), 1.46 (s, 1H, OH), 1.42-1.34 (m, 2H, cyclohexyl-CH2), 1.23-1.16 (m, 2H, cyclohexyl-CH2).

[0257] Example 79 trans-4-({2-[5,5-dioxido-9-(trifluoromethyl)-6H-dibenzo[c,e][1,2]thiazin-6-yl]acetyl}amino)cyclohexyl 4-nitrobenzenesulfonate (SM589) of general formula 8a: To a solution of the appropriate compound SM588 (0.350 g, 0.77 mmol) of formula 8a (Int-4) in dry CHCl (6 mL) was added 4-nitrobenzenesulfonyl chloride (0.355 g, 1.60 mmol), DMAP (0.094 g, 0.77 mmol), and ETN (0.320 mL, 2.30 mmol) at 0° C. The reaction mixture was stirred at room temperature for 2 h, then concentrated in vacuo, poured into ice water, and acidified (pH = 4) with 2 N HCl. The mixture was extracted with CH2Cl2 (3 × 20 mL), and the combined organic layers were washed with brine, dried over Na2SO4, and evaporated to dryness to give a white solid, which was crystallized from EtOH to give SM589 in 38% yield as a white solid: mp 151–152 °C. 1 H-NMR(400MHz, CDCl3):δ 8.35(d,J=8.5Hz,2H,Ar-H),8.29(s,1H,Ar-H),8.12-7.93(m,4H,Ar-H),7.81(t,J=7.0Hz,1H,Ar-H),7.72-7.61(m,2H,Ar-H),7.26(d,J=8.6H) 2H , cyclohexyl-CH2), 1.68-1.59 (m, 2H, cyclohexyl-CH2), 1.52 (s, 1H, OH), 1.32-1.10 (m, 2H, cyclohexyl-CH2).

[0258] Example 80 trans-4-({2-[5,5-dioxido-9-(trifluoromethyl)-6H-dibenzo[c,e][1,2]thiazin-6-yl]acetyl}amino)cyclohexyl 4-aminobenzenesulfonate (SM656) of Formula 8a: To a solution of compound SM589 (0.400 g, 0.63 mmol) in DMF (30 mL) was added Raney / Ni (10% w / w, 0.046 g). The reaction mixture was stirred at room temperature under H bubbling for 2 hours. The mixture was filtered through Celite®, and the filtrate was evaporated to dryness to give a brown solid, which was crystallized from EtOH to give SM656 in 58% yield as a brownish solid. 1 H NMR(400MHz,DMSO-d6):δ 8.45(s,1H,Ar-H),8.37(d,J=7.9Hz,1H,Ar-H),8.05(d,J=7.2Hz,1H,Ar-H),7.90-7.80(m,3H,Ar- H and NH),7.70(t,J=7.6Hz,1H,Ar-H),7.57(d,J=8.4Hz,1H,Ar-H),7.44(d,J=8.8Hz,2H,Ar-H),6.59 (d, J = 8.6 Hz, 2H, Ar-H), 6.19 (s, 2H, NH2), 4.58 (s, 2H, CH2), 4.11-4.23 (m, 1H, cyclohexyl-CH2), 1.69-1.61 (m, 4H, each 2H, cyclohexyl-CH2), 1.39-1.31 (m, 2H, cyclohexyl-CH2), 1.17-1.08 (m, 2H, cyclohexyl-CH2).

[0259] Example 81 2-(3-Acetyl-4-hydroxy-1,1-dioxide-2H-1,2-benzothiazin-2-yl)-N-cyclohexylacetamide (12a) (Scheme 4). A mixture of 11a (0.63 g, 2.11 mmol), prepared according to the literature, cyclohexylamine (0.53 mL, 4.66 mmol), TBTU (1.63 g, 5.08 mmol), and EtN (4 equiv.) in dry THF was reacted at room temperature for 2 h. The reaction mixture was then poured into ice water and acidified (pH = 4) with 2 N HCl to give a precipitate, which was filtered and dried to give 12a (0.75 g, 94%) as a pale-yellow solid.1 H NMR (400 MHz, DMSO-d6): δ 0.80-1.20 and 1.40-1.60 (m, each 5H, cyclohexyl CH2), 2.40 (s, 3H, CH3), 4.00 (s, 2H, NCH2), 7.75-7.85 (m, 4H, Ar-H and CONH), 7.95-8.10 (m, 1H, Ar-H), 15.20 (bs, 1H, OH).

[0260] Example 82 N-Cyclohexyl-2-(3-methyl-5,5-dioxidopyrazolo[4,3-c][1,2]benzothiazin-4(1H)-yl)acetamide (SM879). A mixture of 12a (0.30 g, 0.79 mmol) and hydrazine monohydrate (0.19 mL, 3.96 mmol) was reacted at 60 °C for 1 h. After cooling, the reaction mixture was poured into ice water and acidified (pH = 4) with 2 N HCl to give a precipitate, which was filtered and purified by flash chromatography eluting with CHCl:MeOH 97:3, followed by crystallization from EtOH to give SM879 (0.08 g, 54%) as a white solid. 1 H NMR (400 MHz, CDCl3) δ 1.10-1.20 and 1.25-1.45 (m, each 2H, cyclohexyl CH2), 1.50-1.75 (m, 4H, cyclohexyl CH2), 2.80-2.90 (m, 2H, cyclohexyl CH2), 2.30 (s, 1H, CH3), 3.75 (m, 1H, cyclohexyl CH), 4.05 (s, 2H, NCH2), 6.50 (d , J = 8.1 Hz, 1H, NH), 7.55 (dt, J = 1.2 and 7.8 Hz, 1H, Ar-CH), 7.70 (dt, J = 1.2 and 7.7 Hz, 1H, Ar-CH), 7.80 (dd, J = 0.9 and 7.8 Hz, 1H, Ar-CH), 7.95 (d, J = 7.3 Hz, 1H, Ar-CH), 10.50 (bs, 1H, CONH). HRMS(ESI) m / z [M+H] + C 24 H 31 Calculated for N3O4S: 375.1460, found: 375.1485. LC-MS: Retention time 4.109 min.

[0261] Example 83 Scheme 9: Synthetic procedure for preparing target compound SM886

[0262] [ka]

[0263] N-(4-aminocyclohexyl)-2-[5,5-dioxide-9-(trifluoromethyl)-6H-dibenzo[c,e][1,2]thiazin-6-yl]acetamide (SM886). A mixture of [5,5-dioxide-9-(trifluoromethyl)-6H-dibenzo[c,e][1,2]thiazin-6-yl]acetic acid (7a, Int-3) (0.10 g, 0.28 mmol), trans-1,4-diaminocyclohexane (0.32 g, 2.80 mmol), TBTU (0.12 g, 0.36 mmol), and DIPEA (0.19 mL, 1.12 mmol) in dry DMF (3 mL) was stirred at room temperature for 3 h. The reaction mixture was poured into ice / water and extracted with CHCl (×3). The combined organic layers were washed with brine, dried over Na2SO4, and evaporated to dryness to give a brown oil. After purification by trituration with Et2O, the title compound was obtained in 16% yield as a yellow solid: mp 212-214 °C. 1 H NMR(400MHz,MeOD):δ 1.16-1.29(m,4H,CH2×2),1.87-1.89(m,4H,CH2×2),2.60-2.63(m,1H,CH),3.49-3.54(m,1H,CH),4.66(s,2H,NCH2),7.57(d,J=8.6Hz,1H ,Ar-H),7.73(t,J=7.5Hz,1H,Ar-H),7.82-7.89(m,2H,Ar-H),7.99(d,J=7.8Hz,1H,Ar-H),8.24(d,J=8.0Hz,1H,Ar-H),8.49(s,1H,Ar-H). HRMS(ESI) m / z[M+H] + C 21 H 22 Calculated value of F3N3O3S: 454.1412, measured value: 454.14162.

[0264] Example 84 Scheme 10: Synthetic procedure for preparing target compound SM887

[0265] [ka]

[0266] Dimethyl 3-({[5,5-dioxido-9-(trifluoromethyl)-6H-dibenzo[c,e][1,2]thiazin-6-yl]acetyl}amino)pentanedioate (8a(Int-5)). A stirred mixture of [5,5-dioxido-9-(trifluoromethyl)-6H-dibenzo[c,e][1,2]thiazin-6-yl]acetic acid of formula 7a(Int3) (0.33 g, 0.92 mmol), dimethyl 3-aminopentanedioate (0.19 g, 1.11 mmol), TBTU (0.38 g, 1.19 mmol), and DIPEA (0.64 mL, 3.68 mmol) in CHCl (10 mL) was kept at room temperature for 2 h. The organic solvent was evaporated, and the residue was poured into ice / water and extracted with EtOAc (×3). The combined organic layers were washed with brine, dried over NaSO, and evaporated to dryness to give a brown oil. After purification by flash column chromatography eluting with CHCl / MeOH (98:2), the title compound was obtained in 25% yield as a white solid: mp 126-128 °C. 1 H NMR(400MHz,CDCl3):δ 2.37-2.44(m,4H,CH2×2),3.58(s,6H,OCH3×2),4.52(s,2H,NCH2),4.59-4.64(m,1H,CH),7.17(d,J=8.4Hz,1H,NH),7.40(d,J=8.5Hz,1H,H-7) ,7.65(t,J=7.8Hz,1H,H-3),7.72(d,J=8.5Hz,1H,H-8),7.79(td,J=1.0 and 7.4Hz,1H,H-2),8.02(d,J=8.1Hz,2H,H-1 and H-4),8.27(s,1H,H-10).

[0267] 2-[5,5-Dioxido-9-(trifluoromethyl)-6H-dibenzo[c,e][1,2]thiazin-6-yl]-N-[3-hydroxy-1-(2-hydroxyethyl)propyl]acetamide (SM887). A mixture of dimethyl 3-({[5,5-dioxido-9-(trifluoromethyl)-6H-dibenzo[c,e][1,2]thiazin-6-yl]acetyl}amino)pentanedioate (0.45 g, 0.87 mmol) of formula 8a (Int-5) and NaBH (1.32 g, 35.97 mmol) in dry THF (20 mL) was stirred at reflux for 16 h. The reaction mixture was then cooled to 0 °C, and excess NaBH was quenched by the addition of MeOH (15 mL). The organic solvent was evaporated, and the residue was poured into ice / water and extracted with EtOAc (×3). The combined organic layers were washed with brine, dried over Na2SO4, and evaporated to dryness to give a yellow oil. After purification by flash column chromatography eluting with CHCl3 / MeOH (98:2), the title compound was obtained in 28% yield as a white solid: mp 136-138 °C. 1 H NMR (400 MHz, DMSO-d): δ 1.45-1.58(m,4H,CH2×2),3.29-3.39(m,4H,CH2×2),3.76-3.78(m,1H,CH),4 .30(t,J=5.1Hz,2H,OH×2),4.66(s,2H,NCH2),7.65(d,J=8.5Hz,1H,H-7),7. 76(t,J=7.5Hz,1H,H-3),7.86-7.93(m,2H,Ar-H),7.97(d,J=7.2Hz,1H,Ar-H ),8.01(d,J=8.6Hz,1H,NH),8.43(d,J=7.9Hz,1H,Ar-H),8.60(s,1H,H-10). 13 C NMR(101MHz,DMSO-d6):δ 38.0,44.1,49.7,58.2,121.6,121.7,123.3,124.4(q,J C-F =268.4Hz), 124.7, 125.6(q,J C-F =32.6Hz),127.1,127.2,129.9,130.9,133.2,135.0,141.9,166.2. HRMS(ESI) m / z[M+K] + C 20 H21 Calculated value of F3N2O5S: 497.0760, measured value: 497.0756.

[0268] Example 85 Scheme 11: Synthetic procedure for preparing target compound SM888

[0269] [ka]

[0270] Alternative procedure for the synthesis of [3-fluoro-5,5-dioxide-9-(trifluoromethyl)-6H-dibenzo[c,e][1,2]thiazin-6-yl]acetic acid (7a(Int-1)). A stirred mixture of the compound of formula 6a(Int-1) (1.25 g, 3.10 mmol) in 1N aqueous LiOH (15.5 mL, 15.5 mmol) and dioxane (30 mL) was kept at room temperature for 30 min. The reaction mixture was poured into ice water and acidified (pH = 2) with 2N HCl. The formed precipitate was filtered and dried to give the title compound in 98% yield; melting point 100-102 °C. 1 H NMR(400MHz,CDCl3):δ 4.72(s,2H,NCH2),7.33(d,J=8.4Hz,1H,Ar-H),7.45(td,J=2.5 and 8.1Hz,1H,H-2 ),7.65-7.72(m,2H,Ar-H),7.98(dd,J=4.4 and 8.6Hz,1H,H-1),8.21(s,1H,H-10).

[0271] Dimethyl 3-({[3-fluoro-5,5-dioxide-9-(trifluoromethyl)-6H-dibenzo[c,e][1,2]thiazin-6-yl]acetyl}amino)pentanedioate (8a(Int-6)). A stirred mixture of [3-fluoro-5,5-dioxide-9-(trifluoromethyl)-6H-dibenzo[c,e][1,2]thiazin-6-yl]acetic acid of formula 7a(Int-1) (0.71 g, 1.9 mmol), dimethyl 3-aminopentanedioate (0.40 g, 2.28 mmol), TBTU (0.79 g, 2.47 mmol), and DIPEA (1.32 mL, 7.6 mmol) in CHCl (30 mL) was kept at room temperature for 2 h. The organic solvent was evaporated, the residue was poured into ice / water, the mixture was acidified (pH=4) with 2N HCl, and the mixture was kept under stirring for 10 min until a precipitate was observed, which was filtered to give the title compound in 87% yield as a white solid: mp 153-155°C. 1 H NMR(400MHz,CDCl3):δ 2.60-2.69(m,4H,CH2×2),3.89(s,6H,OCH3×2),4.57(s,2H,NCH2),4.64-4.66(m,1H,CH),7.14(d,J=8.5Hz,1H,NH),7.48(d,J=8.4 Hz, 1H, Ar-H), 7.54 (td, J = 2.3 and 8.4Hz, 1H, Ar-H), 7.75-7.77 (m, 2H, Ar-H), 8.07 (dd, J = 4.7 and 8.9Hz, 1H, Ar-H), 8.26 (s, 1H, Ar-H).

[0272] 2-[3-Fluoro-5,5-dioxide-9-(trifluoromethyl)-6H-dibenzo[c,e][1,2]thiazin-6-yl]-N-[3-hydroxy-1-(2-hydroxyethyl)propyl]acetamide (SM888). A mixture of dimethyl 3-({[3-fluoro-5,5-dioxide-9-(trifluoromethyl)-6H-dibenzo[c,e][1,2]thiazin-6-yl]acetyl}amino)pentanedioate (0.45 g, 0.85 mmol) of Formula 8a (Int-6) and NaBH (1.28 g, 33.81 mmol) in dry THF (15 mL) was stirred at reflux for 30 h. The reaction mixture was then cooled to 0 °C, and excess NaBH was quenched by the addition of MeOH (15 mL). The organic solvent was evaporated, and the residue was poured into ice / water and extracted with EtOAc (x3). The combined organic layers were washed with brine, dried over Na2SO4, and evaporated to dryness to give a yellow oil. After purification by flash column chromatography eluting with cyclohexane / EtOAc (70:30), the title compound was obtained in 9% yield as a white solid; melting point 136-138 °C. 1 H NMR(400MHz,DMSO-d6):δ 1.43-1.58(m,4H,CH2×2),3.27-3.34(m,4H,CH2×2),3.74-3.76(m,1H,CH),4.30(t, J=5.1Hz,2H,OH×2),4.66(s,2H,NCH2),7.69(d,J=8.5Hz,1H,H-7),7.76(td,J=2.7 and and 8.7 Hz, 1H, H-2), 7.84 (dd, J = 2.7 and 8.6 Hz, 1H, H-4), 7.91 (dd, J = 1.6 and 8.5 Hz, 1H, H-8), 8.02 (d, J = 8.6 Hz, 1H, NH), 8.51 (dd, J = 4.4 and 8.3 Hz, 1H, H-1), 8.60 (s, 1H, H-10). 13 C NMR(101MHz,DMSO-d6):δ 38.2,44.3,50.7,58.3,109.1(d,J C-F =25.5Hz), 120.8(d,J C-F =22.1Hz),122.5,123.5,124.5(q,J C-F =274.0Hz), 124.7, 126.1(q,J C-F=33.2Hz),127.2,127.9,130.7(d,J C-F =8.4Hz), 136.7(d,J C-F =7.5Hz), 141.7, 162.3(d,J C-F =252.8Hz),166.4. HRMS(ESI) m / z[M+Na] + C 20 H 20 Calculated value of F4N2O5S: 499.09267, measured value: 499.09354.

[0273] Example 86 N-{1-[(dimethylamino)methyl]propyl}-2-[3-fluoro-5,5-dioxide-9-(trifluoromethyl)-6H-dibenzo[c,e][1,2]thiazin-6-yl]acetamide (SM889). A mixture of [3-fluoro-5,5-dioxide-9-(trifluoromethyl)-6H-dibenzo[c,e][1,2]thiazin-6-yl]acetic acid (7a (Int-1)) (0.30 g, 0.8 mmol), (2-aminobutyl)dimethylamine (0.13 mL, 0.96 mmol), TBTU (0.33 g, 1.04 mmol), and DIPEA (0.56 mL, 3.2 mmol) in CHCl (30 mL) was stirred at room temperature for 1 h. The organic solvent was evaporated, and the residue was poured into ice / water and extracted with EtOAc (×3). The combined organic layers were washed with brine, dried over NaSO, and evaporated to dryness to give a brown oil. After purification by flash column chromatography eluting with CHCl / MeOH (95:5), the title compound was obtained as a light brown solid in 17% yield: mp 167-169 °C. 1H NMR(400MHz,CDCl3):δ 0.88(t,J=7.4Hz,3H,CH2CH3),1.46-1.49(m,1H,CHCH2CH3×1 / 2),1.60-1.62(m,1H,CHCH2CH3×1 / 2),2.2 1-2.23(m,1H,CHCH2N×1 / 2),2.26-2.29(m,1H,CHCH2N×1 / 2),3.89-3.93(m,CH,1H),4.46(d,J=17.5Hz,1H , NCH2×1 / 2), 4.70 (d, J=17.5Hz, 1H, NCH2×1 / 2), 6.46 (d, J=6.0Hz, 1H, NH), 7.51-7.56 (m, 1H, H-2), 7.60 (d, J=8.6Hz, 1H, H-7), 7.73-7.77 (m, 2H, H-4 and H-8), 8.07 (dd, J=4.5 and 8.8Hz, 1H, H-1), 8.26 (s, 1H, H-10). 13 C NMR(101MHz,CDCl3):δ 9.9,25.9,45.7,49.3,51.5,62.5,110.0(d,J C-F =25.3Hz), 120.8, 120.9(d,J C-F =21.2Hz), 122.7(d,J C-F =3.0Hz), 123.7(q,J C-F =273.7Hz),123.9,127.2,127.7(d,J C-F =3.0Hz), 127.8(q,J C-F =33.3Hz), 128.7(d,J C-F =8.0Hz), 135.9(d,J C-F =7.1Hz), 140.5, 162.4(d,J C-F =256.5Hz),166.8. HRMS(ESI) m / z[M+H] + C 21 H 23 Calculated value of F4N3O3S: 474.1474, measured value: 474.14908.

[0274] Example 87 Scheme 12: Synthetic procedure for preparing intermediates of formula 7a (Int-4)

[0275] [ka]

[0276] [3-Methoxy-5,5-dioxide-9-(trifluoromethyl)-6H-dibenzo[c,e][1,2]thiazin-6-yl]acetic acid (7a(Int-4)). A mixture of the compound of formula 6a(Int-2) (0.34 g, 0.76 mmol) in 10% aqueous NaOH (3 mL) and MeOH (3 mL) was stirred at reflux for 1 h. The reaction mixture was poured into ice / water and acidified (pH = 2) with 2 N HCl. The formed precipitate was filtered and dried to give the title compound in 46% yield. Mp 184-186 °C. 1 H NMR(400MHz,DMSO-d6):δ 3.47(s,3H,OCH3),4.27(s,2H,NCH2),7.38-7.41(m,2H,Ar-H),7.55-7.58(m,1H,A r-H),7.76(d,J=7.3Hz,1H,Ar-H),8.31(d,J=8.7Hz,1H,Ar-H),8.45(s,1H,Ar-H).

[0277] Scheme 13: Synthetic procedure for preparing intermediates of formula 8a (Int-7), 8a (Int-8) and 8a (Int-9)

[0278] [ka]

[0279] Example 88 N-Cyclohexyl-2-[3-methoxy-5,5-dioxide-9-(trifluoromethyl)-6H-dibenzo[c,e][1,2]thiazin-6-yl]acetamide (8a (Int-7)). A stirred mixture of 3-methoxy-9-(trifluoromethyl)-6H-dibenzo[c,e][1,2]thiazin-5,5-dioxide (0.19 g, 0.48 mmol) of formula 7a (Int-4), cyclohexylamine (0.07 mL, 0.57 mmol), TBTU (0.20 g, 0.62 mmol), and DIPEA (0.25 mL, 1.91 mmol) in dry CHCl (10 mL) was kept at room temperature for 2 h. The organic solvent was evaporated, and the residue was poured into ice / water. The resulting precipitate was filtered to give the title compound in 58% yield as a white solid: mp 184-185°C. 1 H NMR (400 MHz, CDCl): δ 1.14-1.22(m,4H,CH2×2),1.32-1.41(m,2H,CH2),1.63-1.66(m,2H,CH2),1.86 -1.89(m,2H,CH2),3.87-3.89(m,1H,CH),3.98(s,3H,OCH3),4.55(s,2H,NCH2) ,6.58(d,J=7.3Hz,1H,NH),7.34-7.38(m,2H,H-1 and H-2),7.52(d,J=2.4Hz,1H, H-4),7.70(d,J=8.6Hz,1H,H-8),7.97(d,J=8.8Hz,1H,H-7),8.24(s,1H,H-10).

[0280] Example 89 2-[3-Methoxy-5,5-dioxide-9-(trifluoromethyl)-6H-dibenzo[c,e][1,2]thiazin-6-yl]-N-(tetrahydro-2H-pyran-4-yl)acetamide (8a (Int-8)). A mixture of 3-methoxy-9-(trifluoromethyl)-6H-dibenzo[c,e][1,2]thiazine 5,5-dioxide (7a (Int-4)) (0.45 g, 0.96 mmol), 4-aminotetrahydropyran (0.12 mL, 1.15 mmol), TBTU (0.40 g, 1.25 mmol), and DIPEA (0.67 mL, 3.84 mmol) in dry CHCl (10 mL) was stirred at room temperature for 2 h. The organic solvent was evaporated, and the residue was poured into ice / water. The resulting precipitate was filtered to give the title compound in 55% yield as a white solid: mp 118-120°C. 1 H NMR(400MHz,CDCl3):δ 1.07-1.24(m,2H,CH2),1.29-1.40(m,1H,CH2×1 / 2),1.59-1.65(m,1H,CH2×1 / 2),3.23-3 .42(m,3H,CH2×1 / 2 and CH2),3.60-3.69(m,1H,CH),3.75-3.80(m,1H,CH2×1 / 2),3.93(s,3H ,OCH3),4.63(s,2H,NCH2),7.38-7.40(m,2H,Ar-H and CONH),7.56-7.63(m,1H,Ar-H),7.78 -7.84(m,1H,Ar-H),8.21-8.25(m,1H,Ar-H),8.35-8.33(m,1H,Ar-H),8.50(s,1H,Ar-H).

[0281] Example 90 N-(1-Ethylpropyl)-2-[3-methoxy-5,5-dioxide-9-(trifluoromethyl)-6H-dibenzo[c,e][1,2]thiazin-6-yl]acetamide (8a (Int-9)). A stirred mixture of 3-methoxy-9-(trifluoromethyl)-6H-dibenzo[c,e][1,2]thiazin-5,5-dioxide (0.45 g, 0.96 mmol) of formula 7a (Int-4), 3-aminopentane (0.13 mL, 1.15 mmol), TBTU (0.40 g, 1.25 mmol), and DIPEA (0.67 mL, 3.84 mmol) in dry CHCl (10 mL) was kept at room temperature for 2 h. The organic solvent was evaporated, and the residue was poured into ice / water. The resulting precipitate was filtered to give the title compound in 55% yield as a white solid: mp 151-153°C. 1 H NMR(400MHz,CDCl3):δ 0.73(t,J=6.7Hz,6H,CH3×2),1.24-1.29(m,2H,CH2),1.38-1.44(m,2H,CH2 ),3.39-3.43(m,1H,CH),3.93(s,3H,OCH3),4.65(s,2H,NCH2),7.39-7.42(m ,2H,Ar-H and CONH),7.63(d,J=8.1Hz,1H,Ar-H),7.83(d,J=8.2Hz,1H,Ar-H),7.89(d,J=8.2Hz,1H,Ar-H),8.35(d,J=8.3Hz,1H,Ar-H),8.50(s,1H,Ar-H).

[0282] Scheme 14: Synthetic procedure for preparing target compound SM890

[0283] [ka]

[0284] N-Cyclohexyl-2-[3-hydroxy-5,5-dioxide-9-(trifluoromethyl)-6H-dibenzo[c,e][1,2]thiazin-6-yl]acetamide (SM890). N-Cyclohexyl-2-[3-methoxy-5,5-dioxide-9-(trifluoromethyl)-6H-dibenzo[c,e][1,2]thiazin-6-yl]acetamide 8a (Int-7) (0.13 g, 0.28 mmol) in dry CHCl (8 mL) was added dropwise with 1 M BBr in dry CHCl (0.84 mL, 0.84 mmol) at 0 °C, and the reaction mixture was then kept at 10 °C for 2 h. The mixture was poured into ice / water and extracted with EtOAc (×3). The combined organic layers were washed with brine, dried over NaSO, and evaporated to dryness to give a brown solid. After purification by flash column chromatography eluting with CHCl3 / MeOH (99:1), the title compound was obtained in 24% yield as a little brown solid: mp 236-240°C. 1 H NMR(400MHz,DMSO-d6):δ 1.13-1.20(m,6H,CH2×3),1.51-1.56(m,1H,CH),1.65-1.67(m,4H,CH2×2),4.60(s,2H,NCH2),7.22-7.25(m,2H,Ar-H),7.57(d,J=8. 3Hz,1H,Ar-H),7.80(d,J=7.9Hz,1H,NH),8.06-8.09(m,1H,Ar-H),8.22(d,J=8.3Hz,1H,Ar-H),8.43(s,1H,Ar-H),10.78(bs,1H,OH). 13 C NMR(101MHz,DMSO-d6):δ 24.7,25.5,32.6,48.1,49.8,107.1,120.7,121.6,121.7,122.1(2C),124.5(q,J C-F =272.9Hz), 125.4(q,J C-F =32.8Hz),125.6,129.2,136.2,140.8,158.9,165.5. HRMS(ESI) m / z[M+Na] + C 21 H 21 Calculated value of F3N2O4S: 477.1071, measured value: 477.10749.

[0285] Scheme 15: Synthetic procedure for preparing target compound SM891

[0286] [ka]

[0287] 2-[3-Hydroxy-5,5-dioxide-9-(trifluoromethyl)-6H-dibenzo[c,e][1,2]thiazin-6-yl]-N-(tetrahydro-2H-pyran-4-yl)acetamide (SM891). 2-[3-Methoxy-5,5-dioxide-9-(trifluoromethyl)-6H-dibenzo[c,e][1,2]thiazin-6-yl]-N-(tetrahydro-2H-pyran-4-yl)acetamide 8a (Int-8) (0.25 g, 0.52 mmol) in dry CHCl (6 mL) was added dropwise with 1 M BBr in dry CHCl (2.34 mL, 2.34 mmol) at 0 °C, and the reaction mixture was then kept at 10 °C for 24 h. The mixture was poured into ice / water and extracted with EtOAc (×3). The combined organic layers were washed with brine, dried over NaSO, and evaporated to dryness to give a brown solid. After purification by flash column chromatography eluting with CHCl / MeOH (98:2), the title compound was obtained in 6% yield as a slightly brown solid: mp 226-228 °C. 1 H NMR(400MHz,DMSO-d6):δ 1.30-1.39(m,2H,CH2),1.62-1.65(m,2H,CH2),3.26-3.32(m,2H,CH2O),3.63-3.67(m,1H,CH),3.77-3.80(m,2H,CH2O),4.61(s,2H,NCH2),7.21 -7.24(m,2H,Ar-H),7.58(d,J=8.3Hz,1H,Ar-H),7.81(d,J=8.6Hz,1H,Ar-H),8.21-8.24(m,2H,Ar-H and NH),8.44(s,1H,Ar-H),10.70(bs,1H,OH).

[0288] Scheme 16: Synthetic procedure for preparing target compound SM892

[0289] [ka]

[0290] N-(1-Ethylpropyl)-2-[3-hydroxy-5,5-dioxide-9-(trifluoromethyl)-6H-dibenzo[c,e][1,2]thiazin-6-yl]acetamide (SM892). N-(1-Ethylpropyl)-2-[3-methoxy-5,5-dioxide-9-(trifluoromethyl)-6H-dibenzo[c,e][1,2]thiazin-6-yl]acetamide 8a (Int-9) (0.23 g, 0.72 mmol) in dry CHCl (6 mL) was added dropwise with 1 M BBr in dry CHCl (2.16 mL, 2.16 mmol) at 0 °C, and the reaction mixture was then kept at 10 °C for 2 h. The mixture was poured into ice / water and extracted with EtOAc (×3). The combined organic layers were washed with brine, dried over NaSO, and evaporated to dryness to give a white solid. After purification by flash column chromatography eluting with CHCl3 / MeOH (98:2), the title compound was obtained in 35% yield as a white solid: mp 216-218°C. 1 H NMR(400MHz,CDCl3):δ 0.74(t,J=7.2Hz,6H,CH3×2),1.20-1.28(m,2H,CH2),1.36-1.43(m,2H,CH2 ),3.37-3.43(m,1H,CH),4.62(s,2H,NCH2),7.19-7.24(m,2H,Ar-H and CONH), 7.59(d,J=8.4Hz,1H,Ar-H),7.79(d,J=7.5Hz,1H,Ar-H),7.87(d,J=8.6Hz, 1H,Ar-H), 8.22(d,J=8.8Hz,1H,Ar-H),8.43(s,1H,Ar-H),10.69(s,1H,OH).

[0291] biology Cells and Plasmids Unless otherwise specified, the cell lines used herein were cultured in Dulbecco's Minimal Essential Medium (DMEM, Gibco, #11960-044), 10% heat-inactivated fetal bovine serum (Δ56-FBS), penicillin / streptomycin (Pen / Strep, Corning #20-002-Cl), non-essential amino acids (NEAA, Gibco, #11140-035), and L-glutamine (Gibco, #25030-024). HEK293 cells were obtained from the American College of Cardiology (ATCC) (ATCC CRL-1573). We used a subclone of HEK293 (A23) stably expressing mouse WT, ΔCR, or EGFP-tagged PrP. Cells were passaged into T25 flasks or 100 mm dishes in medium containing 200 μg / ml hygromycin and split every 3–4 days. Cells were not passaged more than 20 times from the original stock. Compounds used in the experiments were resuspended at 30 or 50 mM in DMSO and diluted to create 1000X stock solutions, which were used for serial dilutions. A 1 μl aliquot of each compound dilution point was then added to cells plated in 1 mL of medium without the selection antibiotic. The cloning strategy used to generate cDNAs encoding WT, ΔCR, or EGFP-tagged PrP has been previously described. 20,31,32 The EGFP-PrP construct contains a monomerized version of EGFP inserted after codon 34 of mouse PrP. The identity of all constructs was confirmed by sequencing the entire coding region. All constructs were cloned into the pcDNA3.1(+) / hygro expression plasmid (Invitrogen). All plasmids were transfected using Lipofectamine 2000 (Life Technologies) according to the manufacturer's instructions.

[0292] Drug-based cell assay (DBCA) and MTT assay DBCA was performed as previously described. 24 The same method was used with minor modifications. Briefly, HEK293 cells expressing ΔCR PrP were cultured in 24-well plates at ∼60% confluence on day 1. On day 2, cells were treated with 500 μg / mL Zeocin. Medium (containing fresh Zeocin and / or compound or vehicle) was replaced every 24 h. On day 5, the cell medium was removed, and the cells were incubated in PBS containing 1 mg / mL 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT, Sigma Aldrich, St. Louis, MO) for 30 min at 37°C. The MTT was carefully removed, and the cells were resuspended in 500 μL of DMSO. Values ​​for each well were obtained by measuring at 570 nm using a plate spectrophotometer (Biotek).

[0293] electrophysiology Field Excitatory Post-Synaptic Potentials (EPSPs) were measured in hippocampal slices from 11-week-old C57BL / 6 mice using a multielectrode array (MEA) system. Slices were recorded at a 30-min baseline, and then LTP was induced with tetanic stimulation (3 trains, 500 MHz each) and recorded for another 30 min. Prion synaptic toxicity was induced by incubating slices with 4% w / v lysate of MoRK13 cells chronically infected with the M1000 prion strain for 5 min during baseline. To assess the potential rescuing activity of SM884, the molecule was continuously perfused throughout the entire recording. The percentage of LTP was calculated by considering the mean EPSP amplitude during the last 10 min of recording relative to the mean EPSP amplitude during the last 5 min before tetanic stimulation.

[0294] Immunofluorescence Cells expressing EGFP-PrP were seeded at 12,000 cells / well in CellCarrier-384 Ultra microplates (Perkin Elmer) and grown for approximately 24 hours to obtain a semi-confluent layer (60%). Vehicle (0.1% DMSO, volume equivalent) was used as a negative control. Cells were treated for 24 hours and then fixed by adding methanol-free paraformaldehyde (Thermo Fisher Scientific) to a final concentration of 4% for 12 minutes at room temperature. Plates were then washed twice with PBS and counterstained with Hoechst 33342. Cellular localization of EGFP-PrP was monitored using an Operetta High-Content Imaging System (Perkin Elmer). Imaging was performed in high-field mode using a 20x high-NA objective (0.75). Five magnetic fields were acquired for each well for two channels (380-445 excitation-emission for Hoechst, 475-525 for EGFP and Alexa 488). Image analysis was performed using Harmony software version 4.1 (Perkin Elmer).

[0295] Western blotting Samples were diluted 1:1 with 2X Laemli sample buffer (2% SDS, 10% glycerol, 100 mM Tris-HCl pH 6.8, 0.002% bromophenol blue, 100 mM DTT) and heated at 95°C for 10 min before analysis by SDS-PAGE. Proteins were electrophoretically transferred to a polyvinylidene fluoride (PVDF) membrane and then blocked for 20 min with 5% (w / v) nonfat dry milk in Tris-buffered saline containing 0.05% Tween-20. After incubation with the appropriate primary and secondary antibodies, signals were revealed using enhanced chemiluminescence (Luminata, BioRad) and visualized using a Bio-Rad XRS Chemidoc image scanner (Bio-Rad).

[0296] Preparation of Aβ oligomers Synthetic Aβ(1-42) peptide (Cat. No. KP2107, Karebay Biochem., Rochester, NY) was dissolved in hexafluoro-2-propanol, incubated in a bath sonicator at maximum power for 10 min, centrifuged at 15,000 × g for 1 min, aliquoted, dried, and stored at -80°C. Before use, the dried film was dissolved with DMSO and diluted to 100 μM in F12 medium (Invitrogen, Waltham, MA). Oligomers were obtained by incubating the peptide at 25°C for 16 h. This preparation routinely produces oligomers that elute near the void volume of a Superdex 75 10 / 300 size-exclusion column (GE Healthcare, Little Chalfont, UK) and react with the oligomer-specific antibody A11. Due to the heterogeneity of oligomer size, the final Aβ oligomer concentrations were considered as monomer equivalents.

[0297] Cultured hippocampal neurons Primary neuronal cultures were derived from the hippocampus of 2-day-old mice and cultured as previously described. 11Neurons were seeded on 35 mm dishes (500,000 cells / dish) precoated with 25 μg / mL poly-D-lysine (Sigma P6407) in B27 / Neurobasal-A medium supplemented with 0.5 mM glutamine, 100 units / mL penicillin, and 100 μg / mL streptomycin (all from Invitrogen). Experiments were performed 12 days after seeding. Neurons were pretreated with each candidate compound or control for 20 minutes, followed by exposure to Aβ oligomers (3 μM) for 20 minutes or 3 hours. Triton-insoluble fractions (TIF) were analyzed by immunoblotting using antibodies against phosphorylated SFK (phospho-SFK) (Tyr 416) or Fyn. While phospho-SFK antibodies detect pY416 in several SFKs, previous studies have shown that PrP-dependent activation of kinases is specific to Fyn. Actin was used as a loading control. Subcellular fractionation was performed as previously reported, with minor modifications. Neurons were homogenized using a Potter-Elvehjem homogenizer in 0.32 M ice-cold sucrose buffer (pH 7.4) containing 1 mM HEPES, 1 mM MgCl2, 10 mM NAF, 1 mM NaHCO3, and 0.1 mM PMSF in the presence of protease inhibitors (Complete mini, Roche Applied Science, Penzberg, Germany) and phosphatase inhibitors (PhosSTOP, Roche Applied Science). The sample was centrifuged at 13,000 × g for 15 min to obtain a crude membrane fraction. The pellet was resuspended in a buffer containing 150 mM KCl and 0.5% Triton X-100 and centrifuged at 100,000 × g for 1 h. The final pellet, designated the Triton-insoluble fraction, was rehomogenized in 20 mM HEPES supplemented with protease and phosphatase inhibitors and then stored at −80°C or used directly for further experiments. Protein concentration in each sample was quantified using a Bradford assay (Bio-Rad), and then 5 μg of protein was analyzed by Western blotting.Primary antibodies were as follows: anti-GluN2A and anti-GluN2B (both 1:2000; Invitrogen), anti-GluA1 and anti-GluA2 (both 1:1000; Millipore, Billerica, MA), anti-PSD-95 (postsynaptic density protein 95; 1:2000; Cayman Chemical, Ann Arbor, MI), and anti-actin (1:5000; Millipore). Western blots were analyzed by densitometry using Quantity One software (Bio-Rad). All experiments were repeated with at least four independent culture preparations (n ​​≥ 4).

[0298] Production of recombinant PrP RecHuPrP23-231 was expressed in competent E. coli Rosetta (DE3) bacteria harboring the pOPIN E expression vector containing the wild-type human Prnp construct (N-KKRPKPGGWNTGGSRYPGQGSPGGNRYPPQGGGGWGQPHGGGWGQPHGGGWGQPHGGGWGQPHGGGWGQGGGTHSQWNKPSKPKTNMKHMAGAAAAGAVVGGLGGYMLGSAMSRPIIHFGSDYEDRYYRENMHRYPNQVYYRPMDEYSNQNNFVHDCVNITIKQHTVTTTTKGENFTETDVKMMERVVEQMCITQYERESQAYYQRGSS-C; SEQ ID NO: 1). Bacteria from glycerolate maintained at -80°C were grown overnight in 250 ml Erlenmeyer flasks containing 50 ml of LB broth. The culture was then transferred to two 2L Erlenmeyer flasks. Each flask contained 500 ml of minimal medium supplemented with 3 g / L glucose, 1 g / L NH₄Cl, 1 M MgSO₄, 0.1 M CaCl₂, 10 mg / mL thiamine, and 10 mg / mL biotin. When the culture reached an OD₆ of 0.9–1.2 AU, isopropyl-β-D-1-thiogalactopyranoside (IPTG) was added to induce PrP expression overnight under the same temperature and stirring conditions. The bacteria were then pelleted and lysed, and inclusion bodies were collected by centrifugation and solubilized in 20 mM Tris-HCl, 0.5 M NaCl, 6 M Gd / HCl, pH 8. Although the protein does not contain a His tag, purification of the protein was performed using a histidine affinity column (HisTrap FF crude 5 ml, GE Healthcare Amersham) utilizing the native His residue present in the octapeptide repeat region of PrP. After elution with a buffer containing 20 mM Tris-HCl, 0.5 M NaCl, 500 mM imidazole and 2 M guanidine-HCl (pH = 8), the quality and purity of the protein batches were assessed by BlueSafe (NZYTech, Lisbon) staining after electrophoresis in SDS-PAGE gels.The protein was folded into the PrPC conformation by dialysis against 20 mM sodium acetate buffer (pH = 5). Aggregates were removed by centrifugation. Correct folding was confirmed by CD and protein concentration was confirmed by measuring absorbance at 280 nm. The protein was concentrated using an Amicon centrifugal device and the concentrated solution was stored at -80°C until use.

[0299] dynamic mass redistribution DMR analysis was performed using an EnSight Multimode Plate Reader (Perkin Elmer, Waltham, MA). Immobilization of full-length (residues 23–230) human recombinant PrPc (15 μL / well of 2.5 μM PrPc in 10 mM sodium acetate buffer, pH 5) onto label-free microplates (EnSpire-LFB High Sensitivity Microplates, Perkin Elmer) was achieved by amine coupling chemistry. The interaction of each molecule with PrPc at various concentrations diluted in assay buffer (10 mM PO4, pH 7.5, 2.4 mM KCl, 138 mM NaCl, 0.05% Tween-20) was monitored after a 30-minute incubation at room temperature. All steps were performed using a Zephyr Compact Liquid Handling Workstation (Perkin Elmer). Data acquisition and processing were performed using Kaleido software (Perkin Elmer).

[0300] Statistical analysis of biological data All data were collected and analyzed blindly by two different operators. Statistical analysis, performed with Prism software version 7.0 (GraphPad), included all data points obtained, except for experiments in which negative and / or positive controls did not produce the expected results and were discarded. No tests for outliers were performed. The Kolmogorov-Smirnov normality test was applied (n ≥ 5, where possible). Results are expressed as mean ± standard error unless otherwise specified. In some cases, dose-response experiments were fitted to a four-parameter logistic (4PL) nonlinear regression model using R. 2 Fitting was estimated by calculating the . All data were analyzed with a one-way ANOVA test, including an assessment of data normality, and corrected by a Dunnett post-hoc test. A probability (p) value of <0.05 was considered significant ( * <0.05, ** <0.01, *** <0.001).

[0301] In vitro bone marrow-derived dendritic cells Bone marrow cells were isolated from C57BL / 6 mice as previously described (DOI: 10.1073 / pnas.1619863114). BM was collected from femurs, tibias, and pelvises using a mortar and pestle in 1x PBS supplemented with 0.5% BSA and 2 mM EDTA (MACS buffer), passed through a 70 μm cell strainer, and centrifuged at 1400 rpm for 5 minutes. Erythrocytes were lysed in ACK lysis buffer (0.15 M ammonium chloride, 10 mM potassium carbonate), and debris was removed by gradient centrifugation using Histopaque 1119 (#11191, Sigma-Aldrich) prior to culture. mmCells were grown at 2 × 10 concentrations in Iscove's Modified Dulbecco's Medium (IMDM, #12440053, Thermo Fisher Scientific) supplemented with non-essential amino acids (#11140-035, Thermo Fisher Scientific), 1 mM sodium pyruvate (#11360-070, Thermo Fisher Scientific), 5 mM glutamine (#25030-024, Thermo Fisher Scientific), 50 μM 2-mercaptoethanol (#31350-010, Thermo Fisher Scientific), 100 U / ml penicillin, 100 μg / ml streptomycin (#15140-122, Thermo Fisher Scientific), and 10% FBS (#10270-106, Thermo Fisher Scientific) containing 5% mouse Flt3-L. 6 The cells were resuspended at 200 cells / ml and seeded onto 6-plate tissue culture plates at 5 ml / well for 8–10 days at 37°C. For all culture experiments, loosely adherent and floating cells were collected by gentle pipetting at the indicated time points.

[0302] cDC1 and cDC2 were sorted into complete IMDM and sorted by FACSAria Fusion as pDCs B220+Bst2+, cDC1 B220-CD11c+MHC-II+CD24+CD172α-, and cDC2 B220-CD11c+MHCII+CD24-CD172α+. Cells were sorted to a purity of >99% by post-sort analysis before use in further experiments. It was confirmed to be 5%.

[0303] Induction of EAE All mice used were 12-week-old animals on a C57BL / 6 background. EAE was induced with a mixture of 200 μg of myelin oligodendrocyte glycoprotein fragment MEVGWYRSPFSRVVHLYRNGK (SEQ ID NO: 2; MOG35-55 peptide; #crb1000205n Cambridge Research Biochemicals) and 4 mg / ml Mycobacterium tuberculosis TB H37 Ra (#231141 BD) in incomplete Freund's adjuvant (#263910, BD) at a 1:1 (v / v) ratio. Mice were subcutaneously injected with the MOG / CFA mixture twice (100 μl each time). Mice then received a single intraperitoneal injection of pertussis toxin (#180, List Biological Laboratories) at a concentration of 1 ng / μL in 200 μL of PBS. Mice received a second injection of pertussis toxin at the same concentration two days after the initial EAE induction. Starting on day 10 after EAE induction, mice were orally administered various doses of SM231 dissolved in 1x PBS every other day. Mice were then monitored and scored daily. EAE clinical scores were defined as follows, as previously described (Mayo et al., 2014; Rothhammer et al., 2016): 0—no signs; 1—fully limp tail; 2—hindlimb weakness; 3—hindlimb paralysis; 4—forelimb paralysis; and 5—moribund. Gender differences were not analyzed; only a single sex was used in any set of EAE experiments. Mice were randomly assigned to treatment groups.

[0304] result Identification, characterization, and optimization of SM3. Mutations in the central region of PrP, including artificial deletions or disease-associated point mutations, induce toxic ion channel activity that can be detected in transfected cells by patch clamp techniques. 23,24Cells expressing PrP mutants are also hypersensitive to several cationic drugs commonly used for selection of transfected cell lines, including aminoglycosides and phleomycin analogues. 20 The latter effect was used to establish a novel cellular assay to study mutant PrP-associated toxicity, called the "drug-based cellular assay," or DBCA. 25 Importantly, co-expression of wild-type (WT) PrP suppressed both channel activity and citoxicity, possibly indicating that mutant PrP aberrantly activates signaling pathways normally controlled by PrP. Therefore, DBCA is a unique tool for identifying compounds that can modulate PrP activity. We developed an optimized and scaled-up format of DBCA in 384-well plates, which was later employed to screen tens of thousands of small molecules. 21,22Several compounds were found to suppress the toxicity of mutant PrP without exhibiting detectable toxicity in WT cells. We focused on one of these compounds (designated SM3 [dibenzo[3,4][c,e]thiazine 5,5-dioxide], as shown in Figure 1A). SM3 possesses a drug-like chemical scaffold suitable for optimization and structure-activity relationship (SAR) experiments. Several dozen derivatives (Figure 1C) were designed and synthesized, and their biological activities were tested by DBCA. Three chemical regions of the compound were explored with the dual objectives of improving potency and obtaining SAR information. Using the biological activity of the parent compound SM3 (Figure 1B) as a reference, we evaluated the activity of various derivatives (Figure 2). We noted several important observations about SM3. Chemical modifications performed in the spacer region were unsuccessful. Conversely, substitutions at the C ring improved potency, with the 9-CF3 derivative being the most potent. Branched substituents on the cyclohexyl group were not tolerated, but substituted phenyl rings produced analogs with potency comparable to the reference compound. Taken together, these results provide important SAR information for SM3 and directly suggest chemical schemes for designing further derivatives and functionalized analogs. We also identified a potent derivative, designated SM231, that exhibited DBCA activity in the submicromolar range (Figure 3).

[0305] [Table 11]

[0306] [Table 12]

[0307] [Table 13]

[0308] [Table 14]

[0309] [Table 15]

[0310] SM231 inhibits the synaptic toxic effects of Aβ oligomers. Recent studies have identified a role for PrP in the toxicity of various misfolded oligomers of disease-associated proteins, such as amyloid-β, the accumulation of which underscores the cognitive decline that occurs in Alzheimer's disease. 2,4 The interaction of PrPC with Aβ oligomers unleashes a rapid and toxic signaling pathway that includes activation of metabotropic glutamate receptor 5 (mGluR5), the tyrosine kinase Fyn, and phosphorylation of the NR2B subunit of the NMDA receptor, ultimately leading to dysregulation of receptor function, excitotoxicity, and dendritic spine retraction. 12 To evaluate the effect of SM231 on Aβ-induced Fyn activation, we exposed primary hippocampal neurons to various concentrations of Aβ oligomers for short periods (10, 20, or 60 min). We confirmed that the oligomers induced rapid phosphorylation of Fyn kinase (results at 20 min are shown in Figure 4). Previous findings 26 Similarly, this effect was inhibited by treatment with a PrP-directed compound [termed Fe(III)-TMPyP]. 35 Interestingly, co-incubation with SM231 completely abolished the Aβ effect and restored Fyn phosphorylation to normal levels (Figure 4A). Next, we directly tested the ability of SM231 to inhibit Aβ oligomer-dependent synaptic toxicity. Primary hippocampal neurons were incubated with Aβ oligomers (3 μM) for 3 h. 11Similarly, we observed a decrease in several postsynaptic markers (NMDA receptor subunits, GluN2A and GluN2B, and AMPA, GluA1 and GluA2, and postsynaptic density protein 95, PSD-95) as assessed by Western blotting of Triton-insoluble fractions. These effects were rescued by treatment with the anti-PrPc molecule Fe(III)-TMPyP. Importantly, co-incubation with SM231 for 20 min significantly rescued the levels of all postsynaptic markers. The level of a control protein (actin) was unaffected by either Aβ oligomers or SM231. These data indicate that SM231 inhibits the ability of Aβ oligomers to disrupt PrPc function and activate neurotoxic signaling pathways.

[0311] Chemical optimization of SM231 toward more metabolically stable derivatives. Within this study, further chemical optimization cycles were performed to functionalize positions predicted to positively improve metabolic stability (Figure 5). In particular, the C-3 position of the dibenzothiazine core was functionalized with F and EtO (the SM882 and SM883 derivatives, respectively). In three other molecules, the cyclohexyl was replaced with more stable and hydrophilic groups (morpholine and tetrahydropyran) or ring-opened to form branched chains (SM881, SM884, and SM885). Interestingly, compound SM884 was more potent than SM231 when assayed with DBCA (Figure 6), making these two molecules promising leads for further development.

[0312] SM884 rescues the synaptotoxic effects of prions in mouse brain slices. To test whether SM884 can inhibit prion-induced toxicity in a disease-relevant context, we turned to a recently developed ex vivo toxicity model. 27,28This assay is based on mouse brain slices acutely exposed to brain homogenates from either end-stage mice infected with the mouse-adapted M1000 human prion strain or to lysates from a cell line chronically infected with the human prion strain. We found that SM884 administered at concentrations of 0.1–0.03 μM induced significant (34% and 71%, respectively) rescue of long-term potentiation (LTP; Figure 7). The higher potency detected at the lowest dose may reflect the aggregation tendency of the molecule observed in this experimental condition. These results are also in full agreement with the estimated half-maximal rescuing dose of the compound in cells (0.018 μM), as assessed by DBCA, clearly demonstrating that SM884 can suppress prion-induced synaptic impairment within the low nanomolar concentration range.

[0313] Murine DC1 and DC2 subsets express PrP, and SM231-treated DC2 promotes the expansion of Treg cells in DC-T cell cocultures. Bone marrow-derived dendritic cells were analyzed for PrP expression after stimulation with two different concentrations of SM231, Fe(III)-TMPyP, or vehicle. For this analysis, PrP expression in each DC subset was measured by Western blot using a specific anti-PrP antibody. We found that DC1 and DC2 expressed baseline levels of PrP, which were slightly increased upon SM231 treatment, particularly in DC2 (Figure 8A). To evaluate the inhibitory function of DC1 or DC2 cells after treatment with SM231 or Fe(III)-TMPyP, we performed in vitro cocultures of DCs with naive CD4+ T cells. We found that the priming capacity of conventional DC2 was significantly affected by treatment of DC2 with SM231. Specifically, these cells were able to promote the proliferation of T cells expressing the Treg cell markers FoxP3 and LAP, an effect that required PrP expression in DCs, as it was prevented in DC2 cells transfected with specific PrP siRNA but not control siRNA (Figure 8B). Overall, these data suggest that PrP stimulation may confer tolerogenic functions to DC2s and suppress the default immunogenic program of this subset.

[0314] The compounds SM888 and SM889, like SM231, promote the tolerogenic activity of cDC2. cDC2 cells have been reported to transpresent IL-6, which is essential for myelin peptide-specific encephalitogenic pathogenic TH17 priming in a model of EAE. To assess whether additional derivatives (i.e., SM887, SM888, and SM889) can induce regulatory functions in DC subsets, we performed in vitro cocultures of cDC2 cells with naive ovalbumin (OVA)-specific transgenic CD4+ T cells in the presence of various concentrations of OVA. T cell proliferation was analyzed. We found that cDC2 priming was significantly affected by cDC treatment with SM derivatives, and even more significantly by SM888 and SM889. Specifically, these cells expressed antigen-specific CD4+ T cells. + It was able to suppress the proliferation of T cells, and this effect was more pronounced when the molecule was used at a concentration of 10 μM (FIG. 9).

[0315] Administration of SM231 ameliorates EAE and suppresses inflammatory cytokines in vivo. The present authors investigated whether PrP modulators could have a protective role in this experimental model. Groups of wild-type female C57BL / 6 mice were immunized with the MOG35-55 peptide and intraperitoneally (ip) injected with Fe(III)-TMPyP or SM231 at two doses every other day from day 3 to day 24 post-vaccination. Control mice received vehicle alone. EAE clinical scores were recorded daily over this period (Figure 10A). We found that administration of SM231 (Figure 10B) resulted in reduced disease compared to control mice (P<0.05). At day 25 post-vaccination (d 25 post-vaccination), SM231 reduced white matter demyelination and inflammatory infiltration compared to vehicle-treated controls (Figure 10C). Furthermore, in vivo treatment with SM231 reduced the secretion of proinflammatory cytokines, such as IL-17A and GM-CSF, by CD4+ T cells purified from cervical lymph nodes and restimulated in vitro with MOG. These data also suggest the therapeutic and physiological value of PrP activation in controlling neuroinflammation. These data also suggest that molecules such as SM231 may exert their effects in vivo by regulating potentially inflammatory antigen-presenting cells. Importantly, these data were similar to those obtained with other derivatives of SM231.

[0316] SM231 does not act by directly targeting PrPC. In view of the promising ability of SM compounds to modulate PrPC activity in several experimental settings, we tested the hypothesis that these molecules act by directly targeting the protein. First, we hypothesized that this compound may promote the relocalization of PrPC from the cell surface, a mode of action recently observed for an antiprion phenothiazine derivative (chlorpromazine, CPZ). 29,30HEK293 cells stably expressing EGFP-tagged PrP were treated with various concentrations of SM231, CPZ, or vehicle control, and PrP localization at the cell surface was monitored by imaging techniques (Fig. 11A). The results showed that, consistent with previous data, CPZ induced the relocalization of EGFP-PrP from the cell surface to intracellular compartments in a dose-dependent manner. 30 Conversely, no change was detected for SM231, suggesting that this compound does not exert its effect by inducing relocalization of PrP from the cell surface. Next, we examined whether SM231 can alter PrP expression. HEK293 cells stably expressing wild-type PrP were treated with various concentrations of SM231. Total PrP levels were then assessed in whole-cell lysates by Western blotting (Figure 11B). This examination revealed no differences in PrP expression upon treatment with SM231. Finally, the direct binding of SM231 to recombinant PrP was examined by dynamic mass redistribution (DMR), a biophysical technique previously used to detect the interaction of small molecules with PrP. 26 Fe(III)-TMPyP and chlorpromazine (CPZ) have previously been reported to have high and low affinities for PrP, respectively. 19,30 These two compounds were used as controls. However, no interaction was observed between SM231 and recombinant PrPc, even at the highest concentration (1 mM, Figure 11C). Taken together, these results indicate that SM231 does not act by directly binding to PrPc or by altering its expression or localization.

[0317] FXR inhibitors suppress mutant PrP cytotoxicity. Two FXR agonists, WAY-362450 and fexaramine (the structures of which are shown below), were tested using the DBCA assay.

[0318] [ka]

[0319] HEK293 cells expressing ΔCR PrP were cultured at ∼60% confluence in 24-well plates on day 1. On day 2, cells were treated with 500 μg / mL Zeocin and / or individual FXR agonists at various concentrations (0.03–30 μM) for 72 hours. The medium (containing fresh Zeocin and / or FXR agonists) was replaced every 24 hours. On day 5, cell culture medium was removed, and cells were incubated with 1 mg / mL 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) in PBS for 30 minutes at 37°C to assess cell viability. Interestingly, the FXR agonist WAY-362450 exhibited a 50% (IC) activity in the submicromolar range. 50 ) inhibitory concentrations, dose-dependently rescued ΔCR PrP-dependent cytotoxicity (Figure 12). Importantly, the FXR agonist fexaramine was much less effective, possibly reflecting differential activity of the two agonists on specific FXR isoforms and / or recruiting different coactivators as well. Taken together, these results establish a direct pharmacological link between mutant PrP toxicity and FXR receptor activity and suggest that this receptor may be a target for SM compounds.

[0320] SM231 mediates FXR gene transcriptional activity in mouse hepatocytes. Primary mouse hepatocytes were isolated from 6- to 8-week-old C57Bl6 / J wild-type male mice (obtained from Charles River). 3x 106 Primary hepatocytes were stimulated with increasing concentrations of SM231 or WAY-362450 (potent and selective farnesoid X receptor (FXR) agonists) for 4 or 12 hours. Expression of FXR (nr1h4) and the FXR target gene NrOb2 was assessed by RT-qPCR using specific primers.

[0321] In this experiment, SM231, like the reference agonist WAY-362450, significantly promoted FXR transcriptional activity in these cells, particularly at 3 hours after treatment (Figure 13). This effect was lost 6 hours after activation for both molecules. These data suggest that SM231 can act as an FXR receptor agonist in cells.

[0322] References 1 Chiti, F. & Dobson, CM Protein Misfolding, Amyloid Formation, and Human Disease: A Summary of Progress Over the Last Decade. Annu Rev Biochem 86, 27-68, doi:10.1146 / annurev-biochem-061516-045115 (2017). 2 Selkoe, DJ & Hardy, J. The amyloid hypothesis of Alzheimer's disease at 25 years. EMBO Mol Med 8, 595-608, doi:10.15252 / emmm.201606210 (2016). 3 Walsh, DM et al. Naturally secreted oligomers of amyloid beta protein potently inhibit hippocampal long-term potentiation in vivo. Nature 416, 535-539, doi:10.1038 / 416535a (2002). 4 Lauren, J., Gimbel, D. A., Nygaard, H. B., Gilbert, J. W. & Strittmatter, S. M. Cellular prion protein mediates impairment of synaptic plasticity by amyloid-beta oligomers. Nature 457, 1128-1132, doi:10.1038 / nature07761 (2009). 5 Prusiner, S. B. Prions. Proc Natl Acad Sci U S A 95, 13363-13383, doi:10.1073 / pnas.95.23.13363 (1998). 6 Mallucci, G. et al. Depleting neuronal PrP in prion infection prevents disease and reverses spongiosis. Science 302, 871-874, doi:10.1126 / science.1090187 (2003). 7 Brandner, S. et al. Normal host prion protein necessary for scrapie-induced neurotoxicity. Nature 379, 339-343, doi:10.1038 / 379339a0 (1996). 8 Biasini, E., Turnbaugh, J. A., Unterberger, U. & Harris, D. A. Prion protein at the crossroads of physiology and disease. Trends Neurosci 35, 92-103, doi:10.1016 / j.tins.2011.10.002 (2012). 9 Chung, E. et al. Anti-PrPC monoclonal antibody infusion as a novel treatment for cognitive deficits in an Alzheimer's disease model mouse. BMC Neurosci 11, 130, doi:10.1186 / 1471-2202-11-130 (2010). 10 Gimbel, D. A. et al. Memory impairment in transgenic Alzheimer mice requires cellular prion protein. J Neurosci 30, 6367-6374, doi:10.1523 / JNEUROSCI.0395-10.2010 (2010). 11 Fluharty, B. R. et al. An N-terminal fragment of the prion protein binds to amyloid-beta oligomers and inhibits their neurotoxicity in vivo. J Biol Chem 288, 7857-7866, doi:10.1074 / jbc.M112.423954 (2013). 12 Um, J. W. et al. Alzheimer amyloid-beta oligomer bound to postsynaptic prion protein activates Fyn to impair neurons. Nat Neurosci 15, 1227-1235, doi:10.1038 / nn.3178 (2012). 13 Ferreira, D. G. et al. alpha-synuclein interacts with PrP(C) to induce cognitive impairment through mGluR5 and NMDAR2B. Nat Neurosci 20, 1569-1579, doi:10.1038 / nn.4648 (2017). 14 Aulic, S. et al. alpha-Synuclein Amyloids Hijack Prion Protein to Gain Cell Entry, Facilitate Cell-to-Cell Spreading and Block Prion Replication. Sci Rep 7, 10050, doi:10.1038 / s41598-017-10236-x (2017). 15 Resenberger, U. K., Winklhofer, K. F. & Tatzelt, J. Cellular prion protein mediates toxic signaling of amyloid beta. Neurodegener Dis 10, 298-300, doi:10.1159 / 000332596 (2012). 16 Linden, R. The Biological Function of the Prion Protein: A Cell Surface Scaffold of Signaling Modules. Front Mol Neurosci 10, 77, doi:10.3389 / fnmol.2017.00077 (2017). 17 Manni, G. et al. The cellular prion protein beyond prion diseases. Swiss Med Wkly 150, w20222, doi:10.4414 / smw.2020.20222 (2020). 18 Ballerini, C. et al. Functional implication of cellular prion protein in antigen-driven interactions between T cells and dendritic cells. J Immunol 176, 7254-7262, doi:10.4049 / jimmunol.176.12.7254 (2006). 19 Esterhazy, D. et al. Classical dendritic cells are required for dietary antigen-mediated induction of peripheral T(reg) cells and tolerance. Nat Immunol 17, 545-555, doi:10.1038 / ni.3408 (2016). 20 Massignan, T. et al. A novel, drug-based, cellular assay for the activity of neurotoxic mutants of the prion protein. J Biol Chem 285, 7752-7765, doi:10.1074 / jbc.M109.064949 (2010). 21 Imberdis, T. et al. Identification of Anti-prion Compounds using a Novel Cellular Assay. J Biol Chem 291, 26164-26176, doi:10.1074 / jbc.M116.745612 (2016). 22 Massignan, T. et al. A Small-Molecule Inhibitor of Prion Replication and Mutant Prion Protein Toxicity. ChemMedChem 12, 1286-1292, doi:10.1002 / cmdc.201700302 (2017). 23 Solomon, I. H., Biasini, E. & Harris, D. A. Ion channels induced by the prion protein: mediators of neurotoxicity. Prion 6, 40-45, doi:10.4161 / pri.6.1.18627 (2012). 24 Solomon, I. H. et al. An N-terminal polybasic domain and cell surface localization are required for mutant prion protein toxicity. J Biol Chem 286, 14724-14736, doi:10.1074 / jbc.M110.214973 (2011). 25 Massignan, T., Biasini, E. & Harris, D. A. A Drug-Based Cellular Assay (DBCA) for studying cytotoxic and cytoprotective activities of the prion protein: A practical guide. Methods 53, 214-219, doi:10.1016 / j.ymeth.2010.11.005 (2011). 26 Massignan, T. et al. A cationic tetrapyrrole inhibits toxic activities of the cellular prion protein. Sci Rep 6, 23180, doi:10.1038 / srep23180 (2016). 27 Foliaki, S. T. et al. Prion acute synaptotoxicity is largely driven by protease-resistant PrPSc species. PLoS Pathog 14, e1007214, doi:10.1371 / journal.ppat.1007214 (2018). 28 Foliaki, S. T. et al. Early existence and biochemical evolution characterise acutely synaptotoxic PrPSc. PLoS Pathog 15, e1007712, doi:10.1371 / journal.ppat.1007712 (2019). 29 Biggi, S. et al. Identification of compounds inhibiting prion replication and toxicity by removing PrP(C) from the cell surface. J Neurochem 152, 136-150, doi:10.1111 / jnc.14805 (2020). 30 Stincardini, C. et al. An antipsychotic drug exerts anti-prion effects by altering the localization of the cellular prion protein. PLoS One 12, e0182589, doi:10.1371 / journal.pone.0182589 (2017). 31 Barmada, S., Piccardo, P., Yamaguchi, K., Ghetti, B. & Harris, D. A. GFP-tagged prion protein is correctly localized and functionally active in the brains of transgenic mice. Neurobiol Dis 16, 527-537, doi:10.1016 / j.nbd.2004.05.005 (2004). 32 Ivanova, L., Barmada, S., Kummer, T. & Harris, D. A. Mutant prion proteins are partially retained in the endoplasmic reticulum. J Biol Chem 276, 42409-42421, doi:10.1074 / jbc.M106928200 (2001). 33 Berman, H.M., Battistuz, T., Bhat, T.N., Bluhm, W.F., Bourne, P.E., Burkhardt, K., Feng, Z., Gilliland, G.L., Iype, L., Jain, S., Fagan, P., Marvin, J., Padilla, D.,. Ravichandran, V, Schneider, B., Thanki, N., Weissig, H., Westbrook, J.D., Zardecki, C. The protein data bank, Acta Crystallogr. Sect. D Biol. Crystallogr. 58 (2002) 899e907, https: / / doi.org / 10.1107 / S0907444902003451 34 Friesner, R.A., Banks, J.L., Murphy, R.B., Halgren, T.A., Klicic, J.J., Mainz, D.T., Repasky, M.P., Knoll, E.H., Shelley, M., Perry, J.K., Shaw, D.E., Francis, P., Shenkin P.S. Glide: a new approach for rapid, accurate docking and scoring. 1. Method and assessment of docking accuracy, J. Med. Chem. 47 (2004) 1739e1749, https: / / doi.org / 10.1021 / jm0306430 35 Nicoll, A.J., Clare R. Trevitt, M., Tattum, H., Risse, E., Quarterman, E., Avila Ibarra, A., Wright, C., Graham, S. J., Sessions, R.B., Farrow, M., Waltho, J.P., Clarke, A.R. Collinge, J. Pharmacological chaperone for the structured domain of human prion protein - PNAS, October 12, 2010, vol. 107, no. 41; https: / / doi.org / 10.1073 / pnas.1009062107. 36 Lu, T.T., Makishima, M., Repa, J.J., Schoonjans, K., Kerr, T.A., Auwerx, J., Mangelsdorf, D.J. Molecular basis for feedback regulation of bile acid synthesis by nuclear receptors. Mol Cell. 2000 Sep; 6(3):507-15. 37 Das, B., Venkateswarlu, K., Mahender, G., Mahender, I. A simple and efficient method for α-bromination of carbonyl compounds using N-bromosuccinimide in the presence of silica-supported sodium hydrogen sulfate as a heterogeneous catalyst. Tetrahedron Letters (2005), 46(17), 3041-3044

Claims

1. A compound of the following general formula (I), and any stereoisomers, pharmaceutically acceptable salts, hydrates, and solvates thereof, for use in the treatment of neurodegenerative diseases or immune diseases. 【Chemical 1】 [Wherein, A is a benzene ring or a 5- or 6-membered heteroaromatic ring, B is the following general structure: [Chemical Formula 2] a benzene ring of Alternatively, B is a 5- or 6-membered heteroaromatic ring which may be substituted by one or more substituents, and each of the substituents is independently hydrogen, halogen, nitro, cyano, thiol, C １－４ alkyl, haloalkyl, O-haloalkyl, OC １－４ alkyl, NHC １－４ alkyl, C(=O)C １－６ alkyl, C(=O)OC １－６ alkyl, C(=O)NHC １－４ alkyl, hydroxy, SC １－４ alkyl, OC １－４ alkylamino, and is selected from W is C(=O), C(=S), CH ２ or does not exist, Y is CH ２ , SO ２ , SO, S, C(=O), PO ２ and NR ４ selected from, Z is N or CH, X １ and X ２ is each independently, in each case, hydrogen, halogen, nitro, cyano, thiol, C １－４ alkyl, haloalkyl, O-haloalkyl, OC １－４ alkyl, NHC １－４ alkyl, C(=O)C １－６ alkyl, C(=O)OC １－６ alkyl, C(=O)NHC １－４ alkyl, hydroxy, SC １－４ alkyl, OC １－４ alkylamino, OH, pyrrolidine, piperidine, morpholine, piperazine, N-methylpiperazine, X ３ is hydrogen, methyl, ethyl, isopropyl or benzyl, and X ４ and X ５ each independently in each case is selected from hydrogen, C １－３ alkyl, haloalkyl, halogen, cycloalkyl, amino, hydroxy, cyano, nitro, n is 0, 1, 2, 3, 4, Or, X ３ and X ４ are joined together to form a single bond or C １－４ alkanediyl, and X ３ and X ４ together with the bridging atoms to which they are each attached form a 5- or 6-membered heterocycle, R １ 、 R ２ 、 R ２ａ and R ３ are each independently hydrogen, halogen, nitro, cyano, hydroxy, mercapto, C １－４ alkyl, haloalkyl, O - haloalkyl, OC １－４ alkyl, NHC １－４ alkyl, C(=O)C １－６ alkyl, C(=O)OC １－６ alkyl, C(=O)NHC １－４ alkyl, OC １－４ alkylamino, SC １－４ alkyl, and are selected from R ４ is selected from hydrogen, C １－４ alkyl, C １－４ aminoalkyl, C １－４ hydroxyalkyl, C １－４ nitroalkyl, C １－４ thioalkyl, C １－６ haloalkyl, and Q is C １－８ alkyl, C １－８ alkenyl, cycloalkyl, heterocycloalkyl, aryl ring, heteroaromatic ring, where (1-1) The above-mentioned C １－８ Alkyl may be substituted with hydroxy, OC １－４ alkyl, NHC １－４ alkyl, N(C １－４ alkyl), ２ , NH(C=O)C １－４ alkyl, aryl, heteroaryl, heterocycloalkyl, cycloalkyl, cycloalkenyl, and each of the above-mentioned aryl, heteroaryl, heterocycloalkyl, cycloalkyl, cycloalkenyl may be substituted with methyl, halogen, hydroxy. (1-2) The cycloalkyl and the heterocycloalkyl may each be substituted with OH, OSO ２ R ５ , C １－３ alkyl, NR ６ R ７ ; provided that (i) R ５ is selected from hydrogen, phenyl, heteroaryl, aminophenyl and nitrophenyl, and herein (ii) R ６ and R ７ each independently is selected from H, methyl, C(=O)CH ３ , SO ２ CH ３ and the selection is made from (1-3) The aryl ring or the heteroaromatic ring may each be substituted with one or more substituents, and the substituents are halogen, nitro, cyano, thiol, C １－４ alkyl, haloalkyl, O-haloalkyl, OC １－４ alkyl, NH ２ , NHSO ２ C １－４ alkyl, NHC １－４ alkyl, C(=O)C １－６ alkyl, C(=O)OC １－６ alkyl, C(=O)NHC １－４ alkyl, hydroxy, SC １－４ alkyl, OC １－４ alkylamino.] (provided that the following compound 【Chemical Formula 3】 is not included.)

2. The compound according to claim 1, and any stereoisomers, pharmaceutically acceptable salts, hydrates, and solvates thereof, wherein the neurodegenerative disease or immune disease is Alzheimer's disease, prion disease, multiple sclerosis, autoimmune encephalitis, Parkinson's disease, inflammatory bowel disease or Crohn's disease.

3. A compound of the following general formula (I), and any stereoisomers, pharmaceutically acceptable salts, hydrates, and solvates thereof, for use in the treatment of multiple sclerosis, autoimmune encephalitis or immune diseases. [Chemical Formula 4] [Wherein, A is a benzene ring or a 5- or 6-membered heteroaromatic ring, B is the following general structure: [Chemical Formula 5] a benzene ring of Alternatively, B is a 5- or 6-membered heteroaromatic ring which may be substituted by one or more substituents, and each of the substituents is independently hydrogen, halogen, nitro, cyano, thiol, C １－４ alkyl, haloalkyl, O-haloalkyl, OC １－４ alkyl, NHC １－４ alkyl, C(=O)C １－６ alkyl, C(=O)OC １－６ alkyl, C(=O)NHC １－４ alkyl, hydroxy, SC １－４ alkyl, OC １－４ alkylamino, and is selected from W is C(=O), C(=S), CH ２ or does not exist, Y is CH ２ , SO ２ , SO, S, C(=O), PO ２ and NR ４ selected from Z is N or CH, X １ and X ２ are each independently, in each case, hydrogen, halogen, nitro, cyano, thiol, C １－４ alkyl, haloalkyl, O - haloalkyl, OC １－４ alkyl, NHC １－４ alkyl, C(=O)C １－６ alkyl, C(=O)OC １－６ alkyl, C(=O)NHC １－４ alkyl, hydroxy, SC １－４ alkyl, OC １－４ alkylamino, OH, pyrrolidine, piperidine, morpholine, piperazine, N - methylpiperazine, selected from X ３ is hydrogen, methyl, ethyl, isopropyl or benzyl, X ４ and X ５ each independently is, in each case, selected from hydrogen, C １－３ alkyl, haloalkyl, halogen, cycloalkyl, amino, hydroxy, cyano, nitro, n is 0, 1, 2, 3, 4, and Or, X ３ and X ４ are joined together to form a single bond or C １－４ alkanediyl, and X ３ and X ４ together with the bridging atoms to which they are each attached form a 5- or 6-membered heterocycle, R １ 、 R ２ 、 R ２ａ and R ３ each independently is hydrogen, halogen, nitro, cyano, hydroxy, mercapto, C １－４ alkyl, haloalkyl, O - haloalkyl, OC １－４ alkyl, NHC １－４ alkyl, C(=O)C １－６ alkyl, C(=O)OC １－６ alkyl, C(=O)NHC １－４ alkyl, OC １－４ alkylamino, SC １－４ alkyl, and is selected from R ４ is selected from hydrogen, C １－４ alkyl, C １－４ aminoalkyl, C １－４ hydroxyalkyl, C １－４ nitroalkyl, C １－４ thioalkyl, C １－６ haloalkyl, and Q is C １－８ alkyl, C １－８ alkenyl, cycloalkyl, heterocycloalkyl, aryl ring, heteroaromatic ring, where (3-1) The C １－８ alkyl may be substituted with hydroxy, OC １－４ alkyl, NHC １－４ alkyl, N(C １－４ alkyl) ２ , NH(C=O)C １－４ alkyl, aryl, heteroaryl, heterocycloalkyl, cycloalkyl, cycloalkenyl, and each of the aryl, heteroaryl, heterocycloalkyl, cycloalkyl, cycloalkenyl may be substituted with methyl, halogen, hydroxy (3-2) The cycloalkyl and the heterocycloalkyl may each be substituted with OH, OSO ２ R ５ , C １－３ alkyl, NR ６ R ７ and, where (i) R ５ is selected from hydrogen, phenyl, heteroaryl, aminophenyl and nitrophenyl, and wherein (ii) R ６ and R ７ each independently is H, methyl, C(=O)CH ３ , SO ２ CH ３ selected from, (3-3) The aryl ring or the heteroaromatic ring may each be substituted with one or more substituents, and the substituents are halogen, nitro, cyano, thiol, C １－４ alkyl, haloalkyl, O-haloalkyl, OC １－４ alkyl, NH ２ , NHSO ２ C １－４ alkyl, NHC １－４ alkyl, C(=O)C １－６ alkyl, C(=O)OC １－６ alkyl, C(=O)NHC １－４ alkyl, hydroxy, SC １－４ alkyl, OC １－４ alkylamino.]

4. The compound according to claim 3, and any stereoisomers, pharmaceutically acceptable salts, hydrates, and solvates thereof, wherein the immune disease is inflammatory bowel disease or Crohn's disease.

5. In the general formula (I), (5-1) A is benzene, and / or (5-2) Y is SO ２ and / or (5-3) W is C(=O) or CH ２ and / or (5-4) Z is N, and / or (5-5) X ４ and X ５ is H, The compound according to any one of claims 1 to 4, and any stereoisomers, pharmaceutically acceptable salts, hydrates, and solvates thereof.

6. The following general formula (II): [Chemical Formula 6] (wherein X1, X2, X3, R1, R2, R2a, R3 and Q are the same as in the above formula (I).) The compound according to any one of claims 1 to 4, and any stereoisomers, pharmaceutically acceptable salts, hydrates, and solvates thereof, represented by.

7. The compound according to any one of claims 1 to 4, and any stereoisomers, pharmaceutically acceptable salts, hydrates, and solvates thereof, represented by the following formula [Chemical Formula 7] [Chemical 8] 【Chemical Formula 9】

8. The compound according to claim 3, and any stereoisomers, pharmaceutically acceptable salts, hydrates, and solvates thereof, represented by the following formula 【Chemical 10】

9. The compound according to any one of claims 1 to 4, and any stereoisomers, pharmaceutically acceptable salts, hydrates, and solvates thereof, represented by the following formula 【Chemical 11】

10. A compound represented by the following general formula (III), and any stereoisomers, pharmaceutically acceptable salts, hydrates, and solvates thereof. 【Chemical Formula 12】 [wherein, A is a benzene ring or a 5- or 6-membered heteroaromatic ring, B is a benzene ring having the following general structure: 【Chemical Formula 13】 (wherein, R １ 、 R ２ and R ３ are each independently hydrogen, halogen, nitro, cyano, thiol, C １－４ alkyl, haloalkyl, O-haloalkyl, OC １－４ alkyl, NHC １－４ alkyl, C(=O)C １－６ alkyl, C(=O)OC １－６ alkyl, C(=O)NHC １－６ alkyl, OC １－４ alkylamino, hydroxy, SC １－４ alkyl, and are selected from R ２ａ is hydrogen, CF ３ , F, OH, OC １－４ alkyl, SC １－４ alkyl, OC １－４ alkylamino, and provided that R ２ａ When R is hydrogen or F, ２ and R ３ are each independently selected from F, Cl, Br, CF ３ , OMe, OH, or R １ When R is halogen, ２ａ R is hydrogen, and ２ R and ３ R are each independently hydrogen, halogen, nitro, cyano, thiol, C １－４ alkyl, haloalkyl, O - haloalkyl, OC １－４ alkyl, NHC １－４ alkyl, C(=O)C １－６ alkyl, C(=O)OC １－６ alkyl, C(=O)NHC １－６ alkyl, OC １－４ alkylamino, hydroxy, SC １－４ alkyl, selected from.) is, or Alternatively, B is a 5- or 6-membered heteroaromatic ring which may be substituted by one or more substituents, and each of the substituents is independently hydrogen, halogen, nitro, cyano, thiol, C １－４ alkyl, haloalkyl, O-haloalkyl, OC １－４ alkyl, NHC １－４ alkyl, C(=O)C １－６ alkyl, C(=O)OC １－６ alkyl, C(=O)NHC １－６ alkyl, hydroxy, SC １－４ alkyl, OC １－４ alkylamino, and is selected from W is C(=O), Y is CH ２ , SO ２ , SO, S, C(=O), PO ２ and NR ４ selected from Z is N or CH, X １ and X ２ is, independently in each case, in each case hydrogen, halogen, nitro, cyano, thiol, C １－４ alkyl, haloalkyl, O-haloalkyl, OC １－４ alkyl, NHC １－４ alkyl, C(=O)C １－６ alkyl, C(=O)OC １－６ alkyl, C(=O)NHC １－４ alkyl, hydroxy, SC １－４ alkyl, OC １－４ alkylamino, OH, pyrrolidine, piperidine, morpholine, piperazine, N-methylpiperazine, X ３ is hydrogen, methyl, ethyl, isopropyl or benzyl, and X ４ and X ５ each independently, in each case, is selected from hydrogen, C １－３ alkyl, haloalkyl, halogen, cycloalkyl, amino, hydroxy, cyano, nitro, n is 0, 1, 2, 3, 4, Or, X ３ and X ４ are joined together to form a single bond or C １－４ alkanediyl, and X ３ and X ４ together with the bridging atoms to which each is attached form a 5- or 6-membered heterocycle, R ４ is selected from hydrogen, C １－４ alkyl, C １－４ aminoalkyl, C １－４ hydroxyalkyl, C １－４ nitroalkyl, C １－４ thioalkyl, C １－６ haloalkyl, and Q is C １－８ alkyl, C １－８ alkenyl, cycloalkyl, heterocycloalkyl, aryl ring, heteroaromatic ring, where (10-1) The C １－８ alkyl may be substituted with hydroxy, OC １－４ alkyl, NHC １－４ alkyl, N(C １－４ alkyl), ２ , NH(C=O)C １－４ alkyl, aryl, heteroaryl, heterocycloalkyl, cycloalkyl, cycloalkenyl, and each of the aryl, heteroaryl, heterocycloalkyl, cycloalkyl, cycloalkenyl may be substituted with methyl, halogen, hydroxy. (10-2) The cycloalkyl and the heterocycloalkyl may each be substituted with OH, OSO ２ R ５ , C １－３ alkyl, NR ６ R ７ and, where (i) R ５ is selected from hydrogen, phenyl, heteroaryl, aminophenyl and nitrophenyl, and wherein (ii) R ６ and R ７ each independently is selected from H, methyl, C(=O)CH ３ , SO ２ CH ３ and is selected from (10-3) The aryl ring or the heteroaromatic ring may each be substituted with one or more substituents, and the substituents are halogen, nitro, cyano, thiol, C １－４ alkyl, haloalkyl, O-haloalkyl, OC １－４ alkyl, NH ２ , NHSO ２ C １－４ alkyl, NHC １－４ alkyl, C(=O)C １－６ alkyl, C(=O)OC １－６ alkyl, C(=O)NHC １－４ alkyl, hydroxy, SC １－４ alkyl, OC １－４ alkylamino.]

11. In the general formula (III), (11-1) A is benzene, and / or (11-2) Y is SO ２ and / or (11-3) Z is N, and / or (11-4) X ４ and X ５ is H The compound according to claim 10, and any stereoisomer, pharmaceutically acceptable salt, hydrate, solvate thereof.

12. The compound represented by the general formula (III) is the compound according to claim 10 or 11 represented by the following formula 【Chemical 14】 【Chemical Formula 15】 and any stereoisomer, pharmaceutically acceptable salt, hydrate, solvate thereof.

13. The compound represented by the general formula (III) is the compound according to claim 10 or 11 for medical use, and any stereoisomer, pharmaceutically acceptable salt, hydrate, solvate thereof.

14. The compound according to claim 13, and any stereoisomer, pharmaceutically acceptable salt, hydrate, solvate thereof for use in the treatment of neurodegenerative diseases or immune diseases.

15. The neurodegenerative disease or immune disease is a prion disease, Alzheimer's disease, multiple sclerosis, autoimmune encephalitis, Parkinson's disease, inflammatory bowel disease or Crohn's disease, and the compound according to claim 14, and any stereoisomer, pharmaceutically acceptable salt, hydrate, solvate thereof.

16. A compound for use according to any one of claims 1 to 4, and any stereoisomer, pharmaceutically acceptable salt, hydrate, solvate thereof, which regulates the activity of normal prion protein (PrPC), or the compound according to claim 10 or 11, and any stereoisomer, pharmaceutically acceptable salt, hydrate, solvate thereof.

17. The compound for use according to any one of claims 1 to 4, and any stereoisomer, pharmaceutically acceptable salt, hydrate, solvate thereof, or the compound according to claim 10 or 11, and any stereoisomer, pharmaceutically acceptable salt, hydrate, solvate thereof, which is administered orally or parenterally in a dosage range of 0.001 to 1000 mg per kg of mammalian body weight per day, either as a single dose or in divided doses.

18. A pharmaceutical composition comprising at least one compound as shown in any one of claims 1 to 4, and any of its stereoisomers, pharmaceutically acceptable salts, hydrates, solvates, alone or in combination with at least one further active compound, together with at least one pharmaceutically acceptable excipient, which is a pharmaceutical composition for the use according to any one of claims 1 to 4.

19. A pharmaceutical composition for the use according to claim 18, which is orally administered in the form of tablets or capsules containing 1.0 to 500 milligrams of the active ingredient.

20. A pharmaceutical composition comprising at least one compound as described in claim 10 or 11, and any of its stereoisomers, pharmaceutically acceptable salts, hydrates, solvates, alone or in combination with at least one further active compound, together with at least one pharmaceutically acceptable excipient.

21. The pharmaceutical composition according to claim 20, which is a composition for use in the treatment of neurodegenerative diseases or immune diseases.

22. The pharmaceutical composition according to claim 21, wherein the neurodegenerative disease or immune disease is a prion disease, Alzheimer's disease, multiple sclerosis, autoimmune encephalitis, Parkinson's disease, inflammatory bowel disease or Crohn's disease.

23. A method for synthesizing the compound represented by the general formula (III) according to claim 10 (wherein A is benzene and Y is SO ２ ), which comprises the following steps: (a) A step of reacting a compound of formula 1a with an aromatic or heteroaromatic amine of formula 1b in the presence of a solvent such as dichloromethane and an amine such as pyridine, trimethylamine, diethylisopropylamine to obtain a compound of formula 2a: 【Chemical 16】 (Wherein, R1, R2, R2a, R3, A and B are the same as those in the general formula (III).) (b) Under appropriate conditions, in the presence of a Raney-Nickel catalyst or using SnCl ２ ·2H ２ O, reducing the nitro group of the compound of formula 2a to an amino group by hydrogenation to obtain a compound of formula 3a: 【Chemical 17】 (Wherein, R1, R2, R2a, R3, A and B are the same as those in the general formula (III).) (c) A step of converting Compound 3a into a compound of Formula 5a, including reacting with NaNO ２ , NaOH and HCl under appropriate conditions, and a second step using Cu powder and DMSO as a solvent at room temperature: 【Chemical 18】 (Wherein, R1, R2, R2a, R3, A and B are the same as those in the general formula (III).) (d) A step of converting a compound of formula 5a into a compound of general formula (I) or general formula (II), wherein this reaction comprises at least one of the following steps: (d-1) 5a and the formula: hal-(CH ２ ), ｎ -C(=O)OEt alkylating agent or the formula: hal-(CH ２ ), ｎ -Q alkylating agent (wherein hal is bromine or chlorine, and n and Q are the same as in the general formula (I)), reaction with (d-2) Treatment with an amine of the formula: Q-NHX ３ (wherein Q and X3 are the same as in the general formula (I)) under microwave irradiation and neat conditions (d-3) CH ２ Cl ２ in the presence of a condensing agent such as TBTU and DIPEA or using SOCl ２ as a chlorinating agent, coupling with an amine of the formula: Q-NHX ３ (wherein Q and X3 are the same as in the general formula (I)).

Images