Novel compounds for diagnosing TDP-43 proteinopathy

Abstract

The present invention relates to compounds suitable for imaging TDP-43 (transactivation response (TAR) DNA-binding protein 43 kDa) aggregates. The compounds can be used, for example, to diagnose diseases, disorders, or abnormalities associated with TDP-43 aggregates, or TDP-43 proteinopathies, such as amyotrophic lateral sclerosis (ALS), Alzheimer's disease (AD), frontotemporal dementia (FTD), and limbic-predominant age-related TDP-43 encephalopathy (LATE). JPEG2025534373000233.jpg20170

Description

[Technical Field]

[0001] The present invention relates to a compound suitable for imaging TDP-43 (transactivation response (TAR) DNA-binding protein 43 kDa) aggregates. The compound can be used, for example, to diagnose diseases, disorders, or abnormalities associated with TDP-43 aggregates, or TDP-43 proteinopathies, such as amyotrophic lateral sclerosis (ALS), Alzheimer's disease (AD), frontotemporal dementia (FTD), and limbic-predominant age-related TDP-43 encephalopathy (LATE). The present invention also relates to a method for preparing the compound, a diagnostic composition comprising the compound, a method of using the compound, a kit comprising the compound, and uses thereof. [Background technology]

[0002] Age-related brain disorders, characterized by pathological aggregation of proteins in the CNS (central nervous system (proteinopathies)) and peripheral organs, represent one of the leading causes of disability and mortality worldwide. The best-characterized protein that forms extracellular aggregates is amyloid beta (Abeta) in Alzheimer's disease (AD) and Abeta-related disorders. Aggregation-prone proteins associated with other diseases that cause neurodegeneration include, but are not limited to, tau, alpha-synuclein (a-syn), huntingtin, fusion in sarcoma (FUS), dipeptide repeat proteins (DPRs) generated by abnormal translation of C9orf72 repeat expansions, superoxide dismutase 1 (SOD1), and TDP-43. Diseases associated with TDP-43 aggregates are commonly referred to as TDP-43 proteinopathies, and include, but are not limited to, amyotrophic lateral sclerosis (ALS), frontotemporal lobar degeneration (FTLD), including frontotemporal lobe dementia with TDP-43 pathology (FTLD-TDP, frontotemporal lobar degeneration with TDP-43 inclusions), and limbic-predominant age-related encephalopathy (LATE).

[0003] Introduction to TDP-43 The transactivation response (TAR) DNA-binding protein 43 kDa (TDP-43) is a 414-amino acid protein encoded by the TARDBP gene on chromosome 1 p36.2 (ALS10). TARDBP is composed of six exons (exon 1 is non-coding; exons 2-6 are protein-coding). TDP-43 belongs to the family of heterogeneous nuclear ribonucleoprotein (hnRNP) RNA-binding proteins (Wang et al., Trends in Molecular Medicine, Vol. 14, No. 11, 2008, pp. 479-485; Lagier-Tourenne et al., Human Molecular Genetics, Vol. 19, Review Issue 1, 2010, R46-R64). TDP-43 contains five functional domains (Figure 1 in Warraich et al., The International Journal of Biochemistry & Cell Biology, 42 (2010) pp. 1606-1609): two RNA recognition motifs (RRM1 and RRM2) with two highly conserved hexameric ribonucleoprotein 2 (RNP2) and octameric ribonucleoprotein 1 (RNP1) regions; a nuclear export signal (NES) and nuclear localization signal (NLS) that enable it to shuttle between the nucleus and cytoplasm and transport bound mRNA; and a glycine-rich domain at the C-terminus that mediates protein-protein interactions. TDP-43 is involved in multiple aspects of RNA processing, including transcription, splicing, transport, and stabilization (Buratti and Baralle, FEBS Journal, 277 (2010) pp. 2268-2281). It is a highly conserved, ubiquitously expressed protein with tightly autoregulated expression levels that continuously shuttles between the nucleus and cytoplasm but is normally primarily restricted to the nucleus.In 2006, TDP-43 was identified as the protein that accumulates in the vast majority of cases of frontotemporal lobar degeneration (FTLD) with tau-negative, ubiquitin-positive inclusions (referred to as FTLD-TDP) and in most cases of amyotrophic lateral sclerosis (ALS) (Arai et al., Biochemical and Biophysical Research Communications, 351 (2006) pp. 602-611; Neumann et al., Science, 314, (2006), pp. 130-133).

[0004] Thirty-eight dominant negative mutations in TDP-43 have been identified in patients with sporadic and familial ALS, as well as in patients with inherited FTD (K263E, N267S), primarily located in the glycine-rich domain (Figure 1; Lagier-Tourenne and Cleveland, Cell, 136, 2009, pp. 1001-1004). TDP-43 is inherently prone to aggregation, as shown by sedimentation assays, and this tendency is increased by several ALS-associated TARDBP mutations (Ticozzi et al., CNS Neurol. Disord. Drug Targets, 2010, 9(3), pp. 285-296).

[0005] TDP-43 in neurodegeneration TDP-43 aggregates are also associated with a variety of diseases, including frontotemporal dementia (sporadic or familial, with or without motor neuron disease (MND), with progranulin (GRN) mutations, with TARDBP mutations, with valosin-containing protein (VCP) mutations, linked to chromosome 9p, corticobasal degeneration, frontotemporal lobar degeneration with ubiquitin-positive inclusions, argyrophilic grain dementia, and Pick's disease), amyotrophic lateral sclerosis (sporadic ALS, with TARDBP mutations, with ANG mutations), and amylotrophic lateral sclerosis (ALS). It has been identified in an expanding list of conditions, including but not limited to: Ilzheimer's disease (sporadic and familial), Down's syndrome, familial British dementia, polyglutamine diseases (Huntington's disease and SCA3), hippocampal sclerosis dementia, and myophathies (sporadic inclusion body myositis, inclusion body myopathy with VCP mutations, oculopharyngeal muscular dystrophy with rimmed vacuoles, myofibrillar myopathy with MYOT or DES mutations) (Lagier-Tourenne et al., Human Molecular Genetics, 2010, Vol. 19, Review Issue 1 R46-R64).

[0006] Aggregated TDP-43 from patient brains exhibits several abnormal modifications, including hyperphosphorylation, ubiquitination, acetylation, and C-terminal fragmentation through proteolytic cleavage (Arai et al., Biochemical and Biophysical Research Communications, 351 (2006), pp. 602-611; Neumann et al., Science, 314, (2006), pp. 130-133; Neumann et al., Acta Neuropathol., (2009), pp. 117:137-149; Hasegawa et al., Annals of Neurology, 2008, Vol. 64, No. 1, pp. 60-70; Cohen et al., Nat Commun., 2015, 6:5845). Another hallmark of TDP-43 pathology is the redistribution and accumulation of TDP-43 from the nucleus to the cytoplasm. The hallmark lesions of FTLD-TDP are neuronal and glial cytoplasmic inclusions (neuronal cytoplasmic inclusions (NCIs) and glial cytoplasmic inclusions (GCIs), respectively) and degenerating neurites (DNs), which are immunoreactive for TDP-43, ubiquitin, and p62, but negative for other neurodegenerative disease-associated proteins. Differences in inclusion morphology and its tissue distribution correlate with specific mutations and / or clinical presentations. Four types of TDP-43 pathology have been described by histological methods (Mackenzie and Neumann, J. Neurochem., (2016), 138 (Suppl. 1), pp. 54-70). FTLD-TDP type A cases are characterized by abundant short DNs and small, oval, or crescentic NCIs, primarily in layer II of the neocortex (Mackenzie et al., J. Neurochem., 2016, 138 (Suppl. 1), pp. 54–70, Fig. 2f). Lenticular neuronal intranuclear inclusions (NIIs) are also typically present, but less abundant. Cases with this pathology typically present clinically as either behavioral-type frontotemporal dementia (bvFTD) or primary progressive aphasia, nonfluent / agrammatic type (nfvPPA) and are associated with progranulin (GRN) mutations. Neuropsychiatric symptoms are particularly common in those with underlying GRN or C9orf72 mutations.Type A is the most common type, accounting for the majority of AD cases (Josephs et al., Acta Neuropathol. 2014, 127(3), pp. 441-50; Arai et al., Acta Neuropathol. 2009, pp. 117, 125-136). The TDP-43 neuropathology in most cases of LATE is also similar to that of Type A, which is consistently found in FTD-GRN (Nelson et al., Brain, 2019, pp. 142, 1503-1527). Type B cases show a moderate number of small or granular NCIs in both superficial and deep cortical layers, with relatively few DNs and NIIs (Mackenzie et al., J. Neurochem., 2016, 138(Suppl. 1), pp. 54-70, Figure 2g). The majority of NCIs have a scattered granular morphology and are sometimes referred to as "pre-inclusion bodies." Importantly, some cases also have a background in which the TDP-43 threads and puncta are delicate and small. The majority of cases with the coappearance of FTD and ALS symptoms are found to have FTLD-TDP type B pathology. Type C cases have abundant, long, and tortuous neurites, primarily in the superficial cortical laminae, with few or no NCIs (Fig. 2j in Mackenzie et al., J. Neurochem., 2016, 138 (Suppl. 1), pp. 54–70). This pathology is particularly found in cases presenting with svPPA (primary progressive aphasia, semantic type). FTLD-TDP type D exhibits abundant lenticular intranuclear inclusions (NIIs) and short DNs in the neocortex, with only rare NCIs (Fig. 2k in Mackenzie et al., J. Neurochem., 2016, 138 (Suppl. 1), pp. 54–70). This pattern of pathology is found only in cases with VCP associated with inclusion body myositis.

[0007] TDP-43 in FTD Frontotemporal dementia (FTD) is a clinical term covering a wide range of disorders based on degeneration of the frontal and temporal lobes, a pathological feature known as frontotemporal lobar degeneration (FTLD). FTD is the second most common cause of early degenerative dementia in people under 65 years of age (Le Ber, Revue Neurologique, 169 (2013), pp. 811-819). FTD is manifested by several syndromes, including bvFTD, characterized by changes in personality and behavior; semantic dementia (SD) and progressive non-fluent aphasia (PNFA), characterized by changes in language function; and corticobasal syndrome (CBS), progressive supranuclear palsy syndrome, and motor neuron disease (FTD-MND), characterized by motor dysfunction. Diagnosis of these syndromes is complex, and a definitive conclusion can only be reached through postmortem tissue analysis based on immunohistochemistry to detect aggregated proteins and description of the affected brain regions. With regard to pathological proteinaceous inclusions, approximately 45% of cases show pathological accumulation of misfolded tau, 45% of cases have pathological TDP-43, and a smaller subgroup has aggregates of FUS and other proteins. FTLD-TDP is a pathological term that describes FTD cases with TDP-43 pathology found primarily as cytoplasmic or neuritic protein aggregates in neurons and glia containing misfolded, insoluble, phosphorylated, and cleaved TDP-43.

[0008] TDP-43 in ALS Amyotrophic lateral sclerosis (ALS) is an adult-onset neurodegenerative disorder characterized by the early loss of upper and lower motor neurons. ALS progression is marked by fatal paralysis and respiratory failure, with a disease course of 1 to 5 years from diagnosis to death. In most cases of sporadic ALS, neuropathology is characterized by abnormal cytoplasmic accumulation of TDP-43 in neurons and glia in the primary motor cortex, brainstem motor nuclei, spinal cord, and associated white matter tracts. ALS dementia involves accumulation of TDP-43 in the extramotor neocortex and hippocampus. The role of TDP-43 phosphorylation in ALS patients has been investigated using a phospho-specific antibody that tightly binds to nuclear and cytoplasmic TDP-43 inclusions. Amino acids S379, S403, S404, S409, and S410 have been identified as the major sites of phosphorylation of TDP-43 (Hasegawa et al., Ann Neurol., 2008;64:60-70; Neumann et al., Acta Neuropathol., 2009,117:137-149).

[0009] TDP-43 in LATE Neuropathological changes in limbic-predominant age-related TDP-43 encephalopathy (LATE) (LATE-NC) are defined by a stereotypical TDP-43 proteinopathy in older adults with or without comorbid hippocampal sclerosis pathology. LATE-NC is a common TDP-43 proteinopathy associated with an amnestic dementia syndrome mimicking Alzheimer's disease in retrospective autopsy studies. LATE is distinguished from frontotemporal lobar degeneration with TDP-43 pathology based on its epidemiology (LATE typically affects older subjects) and the relatively restricted neuroanatomical distribution of the TDP-43 proteinopathy. No molecular-specific biomarkers exist for LATE. The discovery of the TDP-43 PET tracer may enable accurate, potentially earlier diagnosis and monitoring of disease progression, facilitating long-term drug efficacy measurements in patients undergoing clinical trials (including as a potential exclusion criterion for Alzheimer's disease clinical trials) and longitudinal studies of LATE clinical and pathological progression (Nelson et al., Brain, 2019, Vol. 142; No. 6, pp. 1503-1527).

[0010] TDP-43 in AD and other diseases TDP-43 pathology occurs in up to 57% of brains of patients with Alzheimer's disease (Josephs KA et al., Acta Neuropathol., 2014;127(6):811-824; Josephs KA et al., Acta Neuropathol., 2014;127(3):441-450; McAleese et al., Brain Pathol., July 2017;27(4):472-479). TDP-43 aggregation is associated with cognitive decline, memory loss, and medial temporal lobe atrophy in AD. TDP-43-positive patients are 10 times more likely to be cognitively impaired at death than TDP-43-negative subjects. TDP-43 appears to represent a secondary or independent pathology that shares overlapping features with AD by targeting the medial temporal lobe. Pathological TDP-43 follows the stereotypical deposition pattern captured by TDP-43 in the AD (TAD) staging scheme: TDP-43 is initially deposited in the amygdala (stage I), followed by the hippocampus, limbic system, temporal lobe, and finally the frontostriatum (stage V) (Josephs KA et al., Acta Neuropathol., 2014;127(6):811-824; Josephs KA et al., Acta Neuropathol., 2014;127(3):441-450).

[0011] Diagnostic methods in FTD and ALS Diagnosis of FTD based on clinical signs is insufficient, especially in the early stage, because the clinical picture may overlap with other diseases.Therefore, the development of sensitive and specific biomarkers that can distinguish between types of pathology within the FTD spectrum is an urgent issue.Such tools will enable better detection and understanding of the specific types of pathology that cause neurodegeneration.Ultimately, this will lead to the development of diagnostic biomarkers that enable more efficient and accurate patient selection for long-term monitoring in clinical research, and will support the development of new therapeutic drugs for ALS and FTD.

[0012] Several approaches have aimed to develop biochemical biomarkers to distinguish different types of FTD pathology. Several studies have shown that TDP-43 concentrations are elevated in the cerebrospinal fluid (CSF) of clinically defined FTD or FTD-MND populations, but significant overlap exists with control or AD subjects, and it remains unclear whether such approaches will prove clinically useful (Foulds et al., Acta Neuropathol., 2008, 116:141-146; Steinacker et al., Arch. Neurol., 2008;65(11):1481-1487). Levels of total tau or Thr181-phosphorylated tau do not distinguish FTLD-tau from controls. A potential diagnostic tool for distinguishing between FTLD-tau and FTLD-TDP is a reduced CSF p-tau181 to tau ratio of less than 0.37 (Hu et al., Neurology., 2013;81(22):1945-1952). Another study showed that CSF phosphorylated tau levels are positively associated with brain tau burden in FTD and may help distinguish TDP-43 proteinopathies from tauopathies (Irwin et al., Ann. Neurol., 2017 Aug;82(2):247-258).

[0013] The development of imaging biomarkers, in parallel with biochemical biomarkers, will enable early and specific detection of pathology in FTD and ALS. The ability to image TDP-43 deposition in the brain would be a significant advance for diagnostics and drug development for FTD, ALS, and other neurodegenerative disorders. Progressive TDP-43 accumulation in the CNS is associated with disease progression, making it an obvious target for the development of novel therapeutics and diagnostic tools to study pharmacodynamics and disease progression. Given the relative novelty of TDP-43 as a target, the development of PET tracers targeting this protein has only just begun. However, the majority of compounds reported to date are not specific for TDP-43, and none of these compounds have demonstrated direct target binding.

[0014] The development of a TDP-43-specific PET tracer faces several challenges, including the low abundance and heterogeneous distribution of the target in patient brains and the lack of standard compounds. To reduce background signal interference resulting from nonspecific off-target binding and to reduce dosing requirements, TDP-43 imaging compounds should bind to the target with high affinity and selectivity. To image TDP-43 aggregates associated with neurological disorders, such as FTD and ALS, imaging compounds must cross the blood-brain barrier and penetrate into relevant brain regions. To target intracellular amyloid-like inclusions, such as TDP-43 aggregates, cell permeability is an additional requirement for imaging compounds. Rapid washout of the compound from the brain (or other target organs) is another prerequisite to avoid compound accumulation in tissues, which may increase the risk of undesirable side effects. [Prior art documents] [Patent documents]

[0015] [Patent Document 1] US 8,932,557 [Patent Document 2] WO2023 / 285661 [Non-patent literature]

[0016] [Non-Patent Document 1] Wang et al., Trends in Molecular Medicine, Vol. 14, No. 11, 2008, pp. 479-485 [Non-patent document 2] Lagier-Tourenne et al., Human Molecular Genetics, 2010, Vol. 19, Review Issue 1, R46-R64 [Non-patent document 3] Figure 1 from Warraich et al., The International Journal of Biochemistry & Cell Biology, 42 (2010), pp. 1606-1609 [Non-patent document 4] Buratti and Baralle, FEBS Journal, 277 (2010) pp. 2268-2281 [Non-patent document 5] Arai et al., Biochemical and Biophysical Research Communications, 351 (2006) pp. 602-611. [Non-patent document 6] Neumann et al., Science, 314, (2006), pp. 130-133 [Non-Patent Document 7] Figure 1; Lagier-Tourenne and Cleveland, Cell, 136, 2009, pp. 1001-1004 [Non-patent document 8] Ticozzi et al., CNS Neurol. Disord. Drug Targets, 2010, 9(3), pp. 285-296 [Non-Patent Document 9] Lagier-Tourenne et al., Human Molecular Genetics, 2010, Vol. 19, Review Issue 1, R46-R64 [Non-Patent Document 10] Neumann et al., Acta Neuropathol., (2009) 117:137-149 [Non-Patent Document 11] Hasegawa et al., Annals of Neurology, 2008, Vol. 64, No. 1, pp. 60-70 [Non-Patent Document 12] Cohen et al., Nat Commun.;2015, 6:5845 [Non-Patent Document 13] Mackenzie and Neumann, J. Neurochem., (2016), 138 (Suppl. 1), pp. 54-70 [Non-Patent Document 14] Figure 2f in Mackenzie et al., J. Neurochem., 2016, 138(Suppl. 1), pp. 54-70 [Non-Patent Document 15] Josephs et al., Acta Neuropathol. 2014, 127(3), pp. 441-50 [Non-Patent Document 16] Arai et al., Acta Neuropathol. 2009, 117, pp. 125-136 [Non-Patent Document 17] Nelson et al., Brain, 2019, 142, pp. 1503-1527 [Non-Patent Document 18] Figure 2g in Mackenzie et al., J. Neurochem., 2016, 138(Suppl. 1), pp. 54-70 [Non-Patent Document 19] Figure 2j in Mackenzie et al., J. Neurochem., 2016, 138(Suppl. 1), pp. 54-70 [Non-Patent Document 20] Figure 2k in Mackenzie et al., J. Neurochem., 2016, 138(Suppl. 1), pp. 54-70 [Non-Patent Document 21] Le Ber, Revue Neurologique, 169(2013), pp. 811-819. [Non-Patent Document 22] Hasegawa et al., Ann Neurol., 2008;64:60-70 [Non-Patent Document 23] Neumann et al., Acta Neuropathol., 2009, 117:137-149 [Non-Patent Document 24] Nelson et al., Brain, 2019, Vol. 142; No. 6, pp. 1503-1527 [Non-Patent Document 25] Josephs KA et al., Acta Neuropathol., 2014;127(6):811-824 [Non-Patent Document 26] Josephs KA et al., Acta Neuropathol., 2014;127(3):441-450 [Non-Patent Document 27] McAleese, J., Brain Pathol., 2017, Jul;27(4):472-479.

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[0017] The object of the present invention was to provide compounds capable of binding to TDP-43 aggregates. In particular, the compounds of the present invention should be useful for identifying and distinguishing patients and patient groups with TDP-43 proteinopathies (e.g., FTD, FTLD-TDP, LATE, and ALS), as well as for distinguishing TDP-43 proteinopathies from other proteinopathies. [Means for solving the problem]

[0018] The present inventors have surprisingly found that compounds having formula (I) can recognize and bind to TDP-43 aggregates. Furthermore, it has been found that the compounds of the present invention exhibit high selectivity for TDP-43 aggregates over common pathologies, such as Abeta and tau, in AD brain homogenates and over a-syn in PD brain homogenates. Furthermore, the compounds of the present invention have been shown to exhibit robust brain uptake and rapid washout in non-human primates, fulfilling the criteria for further development of PET tracers for use in human subjects.

[0019] The present invention is summarized in the appended claims. In particular, the present invention relates to a compound of formula (I):

[0020] [ka]

[0021] (In the formula, n is 1 or 2, R 1 is H, hydroxy(C1-C4)alkyl or F, R 2 is a 5- or 6-membered carbocyclic or heterocyclic ring optionally substituted with F, NH, CN, and / or CH, wherein the heterocyclic ring contains one or more heteroatoms selected from N, O, and S), or a detectably labeled compound, stereoisomer, polymorph, racemic mixture, tautomer, pharmaceutically acceptable salt, prodrug, hydrate, or solvate thereof.

[0022] In another aspect, the present invention provides a diagnostic composition comprising a compound according to the definition of compounds of formula (I) or subformulas thereof as defined herein, and optionally at least one physiologically acceptable carrier, diluent, adjuvant and / or excipient, which may be used for imaging of TDP-43 aggregates, particularly where imaging is performed by positron emission tomography, or for diagnosing a disease, disorder or condition associated with TDP-43 aggregates, particularly where diagnosis is performed by positron emission tomography.

[0023] In another aspect, the present invention provides a method comprising: A method for diagnosing a disease, disorder, or abnormality associated with TDP-43 aggregates in a subject; A method for positron emission tomography (PET) imaging of TDP-43 aggregates in tissue of a subject, A method for detecting, and optionally quantifying, TDP-43 aggregates in tissue of a subject; A method for diagnostic imaging of the brain of a subject; A method for collecting data for diagnosing a disease, disorder, or abnormality associated with TDP-43 aggregates or for diagnosing a TDP-43 proteinopathy; A method for collecting data to determine a disease, disorder, or condition associated with TDP-43 aggregates, or a predisposition to a TDP-43 proteinopathy; A method for collecting data for monitoring the progression of a disease, disorder, or condition associated with TDP-43 aggregates or for monitoring the progression of a TDP-43 proteinopathy in a patient, and A method for collecting data to predict the responsiveness of a patient suffering from a disease, disorder, or abnormality associated with TDP-43 aggregates to treatment with a pharmaceutical agent. The present invention provides a compound according to the definition of a compound of formula (I) or a subformula thereof, which can be used in the following:

[0024] In another aspect, the present invention provides compounds according to the definition of compounds of formula (I) or subformulas thereof, which may also be used as biomarkers of TDP-43 aggregates or TDP-43 proteinopathy biomarkers, as TDP-43 proteinopathy diagnostic agents or tools, or as in vitro analytical standards or in vitro screening tools.

[0025] Another aspect of the present invention provides a method for preparing a compound according to the definition of a compound of formula (I) or a subformula thereof.

[0026] In yet another aspect, the present invention relates to a kit for preparing a radiopharmaceutical preparation, the kit comprising a precursor of a compound of formula (I) or a subformula thereof. [Brief explanation of the drawings]

[0027] [Figure 1][Figure 1a] Graph of saturation binding curve for human FTD sarkosyl-insoluble brain extract. Each point represents the mean ± standard error of the mean (SEM) from two independent experiments. Human FTD type A sarkosyl-insoluble brain extract. [H]-Compound 1. [Figure 1b] Graph of saturation binding curve for human FTD sarkosyl-insoluble brain extract. Each point represents the mean ± standard error of the mean (SEM) from two independent experiments. Human FTD type A sarkosyl-insoluble brain extract. [H]-Compound 7. [Figure 1c] Graph of saturation binding curve for human FTD sarkosyl-insoluble brain extract. Each point represents the mean ± standard error of the mean (SEM) from two independent experiments. Human FTD type A sarkosyl-insoluble brain extract. [H]-Compound 8. [Figure 1d] Graph of saturation binding curve for human FTD sarkosyl-insoluble brain extract. Each point represents the mean ± standard error of the mean (SEM) from two independent experiments. Human FTD type A sarkosyl-insoluble brain extract. [H]-Compound 9 [Figure 1e] Graph of the saturation binding curve for human FTD sarkosyl-insoluble brain extract. Each point represents the mean ± standard error of the mean (SEM) from two independent experiments. Human FTD type A sarkosyl-insoluble brain extract. [H]-Compound 15 [Figure 1f] Graph of the saturation binding curve for human FTD sarkosyl-insoluble brain extract. Each point represents the mean ± standard error of the mean (SEM) from two independent experiments. [H]-Compound 16 [Figure 1g] Graph of the saturation binding curve for human FTD sarkosyl-insoluble brain extract. Each point represents the mean ± standard error of the mean (SEM) from two independent experiments. [H]-Compound 17 [Figure 1h] Graph of the saturation binding curve for human FTD sarkosyl-insoluble brain extract. Each point represents the mean ± standard error of the mean (SEM) from two independent experiments. Human FTD type B sarkosyl-insoluble brain extract. [H]-Compound 1. [Figure 1i] Graph of saturation binding curve for human FTD sarkosyl-insoluble brain extract. Each point represents the mean ± standard error of the mean (SEM) of two independent experiments. Human FTD type B sarkosyl-insoluble brain extract. [H]-Compound 17. [Figure 2][Figure 2a] Microautoradiography images of FTLD-TDP brain tissue. Images of silver grain deposits (A, black arrow) colocalized with pTDP-43 immunostaining (B, white arrow) in FTLD-TDP postmortem human brain tissue. The scale bar is 10 μm. [3H]-Compound 1 in FTLD-TDP type A brain tissue. [Figure 2b] Microautoradiography images of FTLD-TDP brain tissue. Images of silver grain deposits (A, black arrow) colocalized with pTDP-43 immunostaining (B, white arrow) in FTLD-TDP postmortem human brain tissue. The scale bar is 10 μm. [3H]-Compound 9 in FTLD-TDP type A brain tissue. [Figure 2c] Microautoradiography images of FTLD-TDP brain tissue. Figure 2d shows a microautoradiography image of FTLD-TDP brain tissue. Figure 2a shows a colocalized silver granule deposit (A, black arrow) with pTDP-43 immunostaining (B, white arrow) in postmortem human brain tissue from FTLD-TDP. The scale bar is 20 μm. [H]-Compound 17 in FTLD-TDP type A brain tissue. [Figure 2d] shows a microautoradiography image of FTLD-TDP brain tissue. Figure 2b shows a colocalized silver granule deposit (A, black arrow) with pTDP-43 immunostaining (B, white arrow) in postmortem human brain tissue from FTLD-TDP. [H]-Compound 1 in FTLD-TDP type B brain tissue. [Figure 2e] shows a microautoradiography image of FTLD-TDP brain tissue. Figure 2c shows a colocalized silver granule deposit (A, black arrow) with pTDP-43 immunostaining (B, white arrow) in postmortem human brain tissue from FTLD-TDP. The scale bar is 20 μm. [3H]-Compound 17 in FTLD-TDP type B brain tissue. [Figure 3][Figure 3a] Classical autoradiography images of [3H]-Compound 1 in FTLD-TDP or control brain tissue. Autoradiography images in control, FTLD-TDP type A, or FTLD-TDP type B brain tissue. [Figure 3b] Classical autoradiography graph of [3H]-Compound 1 in FTLD-TDP or control brain tissue. Images of total binding (Total) and nonspecific binding (Self-Block) for each tissue type are shown. Specific binding (Total-Nonspecific) in control, FTLD-TDP type A, and FTLD-TDP type B. Statistical analysis was performed using ordinary one-way ANOVA with Tukey's correction for multiple comparisons. **p>0.01, *p<0.05. [Figure 3c] Classical autoradiography graph of [3H]-Compound 1 in FTLD-TDP or control brain tissue. Graph of saturation binding curve in FTLD-TDP type A by classical autoradiography. Specific binding of [3H]-Compound 1 is shown. [Figure 4] [Figure 4a] Classical autoradiography images of [3H]-Compound 17 in FTLD-TDP or control brain tissue. Autoradiography images in control, FTLD-TDP type A, or FTLD-TDP type B brain tissue. [Figure 4b] Classical autoradiography graph of [3H]-Compound 17 in FTLD-TDP or control brain tissue. Images of total binding (Total) and nonspecific binding (Self-Block) for each tissue type are shown. Specific binding (Total-Nonspecific) in control, FTLD-TDP type A, and FTLD-TDP type B. Statistical analysis was performed using ordinary one-way ANOVA with Tukey's correction for multiple comparisons. **p>0.01, *p<0.05. [Figure 4c] Classical autoradiography graph of [3H]-Compound 17 in FTLD-TDP or control brain tissue. Saturation binding curve in FTLD-TDP type A by classical autoradiography. Specific binding of [3H]-compound 17 is shown. [Figure 5][Figure 5a] Graph of the binding specificity of [3H]-Compound 1 in extracts derived from human brain. Saturation binding curves of [3H]-Compound 1 and [3H]-Abeta standard in AD brain homogenate. [Figure 5b] Graph of the binding specificity of [3H]-Compound 17 in extracts derived from human brain. Saturation binding curves of [3H]-Compound 17 and [3H]-Abeta standard in AD brain homogenate. [Figure 5c] Graph of the binding specificity of [3H]-Compound 1 in extracts derived from human brain. Saturation binding curves of [3H]-Compound 1 and a-syn standard in insoluble fractions derived from PD brain. [Figure 5d] Graph of the binding specificity of [3H]-Compound 17 in extracts derived from human brain. Saturation binding curves of [3H]-Compound 17 and a-syn standard in insoluble fractions derived from PD brain. [Figure 5e] Graph of the binding specificity of [3H]-Compound 1 in extracts derived from human brain. Saturation binding curves of [3H]-Compound 1 and [3H]-tau standard in tau PHFs derived from AD brain. [Figure 5f] Graph of binding specificity of [3H]-Compound 17 in extracts derived from human brain. Saturation binding curves of [3H]-Compound 17 and [3H]-tau standard in tau PHFs derived from AD brain. [Figure 6] [Figure 6a] Autoradiographic imaging of target engagement in AD brain tissue sections. Classical autoradiography of [3H]-Compound 1 in AD brain tissue sections rich in Abeta- and tau. Scale bar 2 mm. [Figure 6b] Autoradiographic imaging of target engagement in AD brain tissue sections. Classical autoradiography of [3H]-Compound 17 in AD brain tissue sections rich in Abeta- and tau. Scale bar 2 mm. [Figure 6c] Microautoradiography of [3H]-Compound 1 in AD postmortem human brain tissue containing pathological tau aggregates. [Figure 6d] Microautoradiography of [3H]-Compound 17 in AD postmortem human brain tissue containing pathological tau aggregates. [Figure 7][Figure 7a]iv Non-human primate PK graph in monkey whole brain using [18F]-Compound 1. [Figure 7b]iv Non-human primate PK graph in monkey whole brain using [18F]-Compound 17.

[0028] definition Unless otherwise defined, the following definitions apply within the meaning of this application and, where appropriate, terms used in the singular also include the plural and vice versa.

[0029] The compounds of the present invention may have one or more optically active carbons that can exist as racemates and racemic mixtures, stereoisomers (including diastereomeric mixtures and individual diastereoisomers, enantiomeric mixtures and single enantiomers, mixtures of conformers and single conformers), tautomers, atropoisomers, and rotamers. All isomeric forms are included in the present invention. Compounds described herein that contain olefinic double bonds include E and Z geometric isomers. The present invention also includes all salt forms, such as pharmaceutically acceptable salts, polymorphs, hydrates, solvates, prodrugs, and mixtures thereof. Unless otherwise specified, the term "compound of Formula (I)" or "compound of the present invention" refers to a compound of Formula (I), or a detectably labeled compound thereof, stereoisomer, polymorph, racemic mixture, tautomer, pharmaceutically acceptable salt, prodrug, hydrate, or solvate, or mixtures thereof. Unless otherwise specified, the term "compounds of formula (I)" or "compounds of the invention" refers to compounds of formula (I) and subformulas thereof, as well as isotopically labeled compounds ( 18 F and 3 The term "compound of formula (I)" or "compound of the invention" refers to a compound as defined in any one of the embodiments mentioned herein below.

[0030] The term "polymorph" refers to various crystalline structures of the compounds of the present invention. This may include, but is not limited to, crystalline forms (and amorphous materials) and all crystalline lattice forms. Salts may also be crystalline and may exist as more than one polymorph.

[0031] Solvates, hydrates and anhydrous forms of the salts are also included in the present invention. The solvent contained in the solvate is not particularly limited and may be any pharmaceutically acceptable solvent. Examples include C 1~4 Includes alcohol (e.g., methanol or ethanol).

[0032] "Pharmaceutically acceptable salts" are defined as derivatives of the compounds of the present invention where the parent compound is modified by making acid or base salts thereof. Examples of pharmaceutically acceptable salts include, but are not limited to, inorganic or organic acid salts of basic residues, such as amines; alkali or organic salts of acidic residues, such as carboxylic acids; and the like. 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, as well as salts prepared from 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. Pharmaceutically acceptable salts of the compounds of formula (I) 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 the appropriate base or acid in water or an organic solvent, or in a mixture of both. Organic solvents include, but are not limited to, non-aqueous media such as ether, ethyl acetate, ethanol, isopropanol, or acetonitrile. A list of suitable salts can be found in Remington's Pharmaceutical Sciences, 18th Edition, Mack Publishing Company, Easton, PA, 1990, page 1445, the disclosure of which is hereby incorporated by reference. Typically, pharmaceutically acceptable salts are salts of amine residues in the compounds of the present invention.

[0033] The compounds of the invention may also be provided in the form of a prodrug, ie, a compound that is metabolized in vivo to an active metabolite.

[0034] A "patient" or "subject" in the present invention is typically an animal, particularly a mammal, more particularly a human and a mouse, and even more particularly a human.

[0035] A "diagnostic composition" is defined herein as a composition comprising a compound of the invention in a form suitable for administration to a patient, the patient being, for example, a mammal, eg, a human.

[0036] "TDP-43 aggregates" are TDP-43-positive aggregates enriched in TDP-43 multimers. They can be found as intracellular deposits in a series of diseases known as TDP-43 proteinopathies, specifically amyotrophic lateral sclerosis (ALS), Alzheimer's disease (AD), frontotemporal dementia (FTD), and limbic-predominant age-related TDP-43 encephalopathy (LATE). TDP-43 aggregates can be found in the following morphologies: small, oval, or crescentic neuronal cytoplasmic inclusions (NCIs), lens-shaped neuronal intranuclear inclusions (NIIs), glial cytoplasmic inclusions (GCIs), degenerating neurites (DNs), and long, tortuous neurites. In pathological aggregates, TDP-43 often exhibits a significant increase in post-translational modifications, such as phosphorylation, ubiquitination, acetylation, sumoylation, and proteolytic cleavage generating C-terminal fragments.

[0037] The "preclinical state" of a disease is defined as a phase of the disease in which disease-related changes at the molecular level have not yet resulted in a noticeable clinical picture in the patient.

[0038] The "clinical state" of a disease is defined as the phase of the disease in which disease-related changes at the molecular level have given rise to a noticeable clinical picture in the patient.

[0039] The term "diagnosing" or "diagnosis" generally refers to the process or act of recognizing, deciding on, or concluding on a disease or condition in a patient based on symptoms and signs and / or from the results of a diagnostic procedure.

[0040] A "normal control value" is determined by performing each method on multiple healthy subjects, measuring the amount of compound bound to TDP-43 aggregates, if any, in each healthy subject, and calculating the average value.

[0041] A "healthy control subject" or "healthy subject" is an individual who shows no clinical evidence of a neurodegenerative disease. The individual must meet the following criteria: Male and female subjects who are healthy and have no clinically relevant findings on physical examination. No family history of TDP-43 proteinopathy, TDP-43 aggregate formation, or other early-onset neurological disorders associated with dementia. · No personal history of clinically significant neurological and / or psychiatric disorders. · Currently have no signs or symptoms of neurological deficits, e.g., cognitive impairment or motor deficits.

[0042] A "preclinical control value" is determined by performing the respective method on multiple subjects under preclinical conditions, measuring the amount of compound bound to TDP-43 aggregates, if any, in each subject, and calculating the average value.

[0043] A "clinical control value" is determined by performing each method on multiple subjects under clinical conditions, measuring the amount of compound bound to TDP-43 aggregates, if any, in each subject, and calculating the average value.

[0044] The term "predicting" or "prediction" generally refers to the advance pronouncement, indication, or foretelling of a disease or condition in a patient who does not have the disease, disorder, or disorder. For example, predicting a disease, disorder, or disorder in a patient may indicate the likelihood, chance, or risk that the patient will develop the disease, disorder, or disorder, for example, within a certain time period or by a certain age.

[0045] Detectable labels include suitable isotopes, such as radioisotopes, particularly positron- or gamma-emitters; 2H, 3 H, 18 F, 123 I, 124 I, 125 I, 131 I, 11 C. 13 N, 15 O. 99m Tc and 77 Br, preferably 2 H, 3 H, 11 C. 13 N, 15 O, and 18 F, more preferably 2 H, 3 H and 18 F, even more preferably 3 H and 18 F, most preferably 18 Contains F.

[0046] The terms "Hal", "halogen" or "halo" mean F, Cl, Br or I, particularly Br or I, more particularly Br.

[0047] The term "carbocyclic" refers to a 5- or 6-membered carbocyclic ring, including, but not limited to, any 5- or 6-membered saturated or unsaturated carbocyclic ring. Unsaturated carbocyclic rings include, but are not limited to, aromatic rings. Examples of 5- or 6-membered carbocyclic rings include, for example, phenyl, cyclopentyl, cyclohexyl, cyclopentenyl, and cyclohexenyl. Phenyl is preferred.

[0048] The term "heterocyclic ring" refers to a stable 5- or 6-membered heterocyclic ring, including, but not limited to, any 5- or 6-membered saturated or unsaturated heterocyclic ring. Unsaturated heterocyclic rings include, but are not limited to, aromatic rings. The heterocyclic ring contains one or more heteroatoms (e.g., one or two heteroatoms) selected from N, O, and S. The heteroatom is preferably N or O, more preferably N. Examples of 5- or 6-membered heterocyclic rings include, for example, pyridinyl, pyrimidinyl, pyrrolyl, pyrrolidinyl, furanyl, tetrahydrofuranyl, thiophenyl, imidazolidinyl, pyrazolidinyl, imidazolyl, pyrazolyl, oxathiolidinyl, isoxthiolidinyl, oxathiolyl, isoxathiolyl, thiazolidinyl, isothiazolidinyl, thiazolyl and isothiazolyl.

[0049] The term "leaving group" (LG) as used herein means any leaving group, an atom or group of atoms that can be replaced by another atom or group of atoms. Examples are shown, for example, in Synthesis (1982), pp. 85-125, Table 2; Carey and Sundberg, Organische Synthese (1995), pp. 279-281, Table 5.8; or Netscher, Recent Res. Dev. Org. Chem., 2003, pp. 7, 71-83, Schemes 1, 2, 10 and 15, etc. (Schubiger PA, Friebe M., Lehmann L. (eds.), PET-Chemistry - The Driving Force in Molecular Imaging, Springer, Berlin Heidelberg, pp. 15-50; Coenen, Fluorine-18 Labeling Methods: Features and Possibilities of Basic Reactions (2006), explicitly Scheme 4, p. 25; Scheme 5, p. 28; Table 4, p. 30; Figure 7, p. 33). Preferably, the "leaving group" (LG) is C 1~4 Alkyl sulfonate, C6~10 Preferably, the leaving group (LG) is selected from arylsulfonate, arylsulfonate, or nitro. More preferably, the leaving group (LG) is mesylate, tosylate, nosylate, or nitro. Even more preferably, the leaving group (LG) is mesylate or nitro.

[0050] As used herein, the term "detecting" encompasses quantitative and / or qualitative detection.

[0051] The compounds of the invention may be used as analytical standards or in vitro screening tools.

[0052] For example, unlabeled compounds of formula (I) according to the invention can be used as analytical standards for quality control, and the corresponding labeled compounds of the invention, e.g., the corresponding compounds of formula (IF) or (IF'), can be used as analytical standards for quality control. 18 The F-labeled compound can be released. This quality control is performed by an in vitro method.

[0053] The compounds of the present invention can be used as in vitro screening tools to characterize tissues that have tau pathology and to test compounds that target tau pathology in such tissues.

[0054] The preferred definitions provided in the "Definitions" section apply to all of the embodiments described below unless otherwise specified. Various embodiments of the invention are described herein, and it is recognized that the features specified in each embodiment may be combined with other specified features to obtain further embodiments of the invention. DETAILED DESCRIPTION OF THE INVENTION

[0055] Various embodiments of the invention are described herein, and it is recognized that the features specified in each embodiment may be combined with other specified features to yield further embodiments of the invention.

[0056] It is understood that all definitions given for formula (I) apply to all of its subclasses, including formulas (IH), (IF), (II), (III), and (IV).

[0057] In a first aspect, the present invention provides a compound of formula (I)

[0058] [ka]

[0059] (In the formula, n is 1 or 2, R 1 is H, hydroxy(C1-C4)alkyl or F, R 2 is a 5- or 6-membered carbocyclic or heterocyclic ring optionally substituted with F, NH, CN and / or CH, the heterocyclic ring containing one or more heteroatoms selected from N, O and S, preferably N. or a detectably labeled compound, stereoisomer, polymorph, racemic mixture, tautomer, pharmaceutically acceptable salt, prodrug, hydrate or solvate thereof.

[0060] In one embodiment, the present invention provides a compound of formula (I)

[0061] [ka]

[0062] (In the formula, n is 1 or 2, R 1 is H or F, R 2 is a 5- or 6-membered carbocyclic or heterocyclic ring optionally substituted with F, NH, CN and / or CH, the heterocyclic ring containing one or more heteroatoms selected from N, O and S, preferably N. or a detectably labeled compound, stereoisomer, polymorph, racemic mixture, tautomer, pharmaceutically acceptable salt, prodrug, hydrate or solvate thereof.

[0063] The present invention relates to compounds of formula (I) wherein n is 1 or 2. In a preferred embodiment, n is 1. In another preferred embodiment, n is 2.

[0064] The present invention is 1 is H, hydroxy(C1-C4)alkyl or F. In one embodiment, R 1 is H or F. In a preferred embodiment, R 1 is H. In another preferred embodiment, R 1 is F. In another preferred embodiment, R 1 is hydroxy(C1-C4)alkyl. Preferred examples of hydroxy(C1-C4)alkyl are hydroxymethyl or hydroxyethyl, more preferably hydroxymethyl.

[0065] In a preferred embodiment, R 1 is selected from the group consisting of F, H and —CH—OH.

[0066] The present invention is 2 is a 5- or 6-membered carbocyclic or heterocyclic ring optionally substituted with F, NH, CN and / or CH, and a heterocyclic ring refers to a compound of formula (I) containing one or more heteroatoms selected from N, O and S.

[0067] Preferably, R 2 teeth, a 5- or 6-membered carbocyclic ring, preferably an aryl ring, optionally substituted with F, NH2, CN and / or CH3; a 5-membered heteroaryl ring optionally substituted with F, NH2, CN and / or CH3 and containing one or more heteroatoms selected from N, O and S, or a 6-membered heteroaryl ring optionally substituted with F, NH2, CN and / or CH3 and containing 1 or 2 heteroatoms selected from O, N and S; is.

[0068] R 2 Preferred examples of the ring are given in the definitions section above. Preferably, R 2 is phenyl, pyridyl, pyrimidinyl, pyrazolyl, pyridazinyl, thiazolyl or isothiazolyl (e.g., phenyl, pyridyl, pyrimidinyl, pyrazolyl), any of which may be optionally substituted by F, NH2, CN and / or CH3, for example by F, NH2 and / or CH3.

[0069] R 2 The ring may be optionally substituted at any available position with F, NH, CN, and / or CH (e.g., F, NH, and / or CH). In certain embodiments, the 5- or 6-membered carbocyclic or heterocyclic ring is substituted with one or more of F, NH, CN, and / or CH (e.g., F, NH, and / or CH).

[0070] In one embodiment, the present invention provides a compound comprising R 2 is a 5- or 6-membered carbocyclic or heterocyclic ring optionally substituted with F, NH and / or CH, and the heterocyclic ring contains one or more heteroatoms selected from N, O and S. In a preferred embodiment, R 2 is a 5- or 6-membered carbocyclic or heterocyclic ring optionally substituted at any available position with F, NH2 and / or CH3. Preferably, the 5- or 6-membered carbocyclic or heterocyclic ring is substituted with one or more of F, NH2 and / or CH3.

[0071] In one embodiment, the present invention provides a compound comprising R 2 , but optionally, F, NH 2、In a preferred embodiment, R is a 5- or 6-membered carbocyclic or heterocyclic ring optionally substituted with CN, and / or CH, wherein the heterocyclic ring contains one or more heteroatoms selected from N, O and S. 2 Optionally, F, NH, in any available position. 2、 In another embodiment, the 5- or 6-membered carbocyclic or heterocyclic ring is substituted with F, NH 2、 Substituted with one or more of CN and / or CH3.

[0072] In one preferred embodiment, phenyl can be optionally substituted with F, NH2 and / or CN, preferably with CN and NH2, or with F and NH2. In another preferred embodiment, pyrazolyl can be optionally substituted with CH3. In another preferred embodiment, pyridyl can be optionally substituted with F or NH2. In another preferred embodiment, pyridyl is unsubstituted. In another more preferred embodiment, pyrimidinyl is unsubstituted. In another more preferred embodiment, isothiazolyl or thiazolyl is unsubstituted. In another more preferred embodiment, pyrazinyl is unsubstituted.

[0073] In a preferred embodiment, the present invention relates to a compound of formula (I) During the ceremony, R 2 teeth,

[0074] [ka]

[0075] (In the formula, R 3 is F and R 4 is NH2 and R 7 is H and R 8 is H, R 3 is NH2 and R 4 is F and R 7 is H and R 8is H, R 3 is CN and R 4 is NH2 and R 7 is H and R 8 is H, R 3 is H and R 4 is NH2 and R 7 is H and R 8 is CN, R 3 is H and R 4 is NH2 and R 7 is H and R 8 is F, or R 3 is H and R 4 is NH2 and R 7 is H and R 8 is CN).

[0076] In another preferred embodiment, the present invention relates to compounds of formula (I), wherein R 2 teeth

[0077] [ka]

[0078] (X is N and R 5 is H or CH3). In a preferred embodiment, R 5 is CH3.

[0079] In another preferred embodiment, the present invention relates to compounds of formula (I), wherein R 2 teeth,

[0080] [ka]

[0081] (In the formula, R 9 is selected from H, F, CH3 and NH2, preferably H, or R 2 teeth,

[0082] [ka]

[0083] and or R 2 teeth,

[0084] [ka]

[0085] and or R 2 teeth,

[0086] [ka]

[0087] and or R 2 teeth,

[0088] [ka]

[0089] and or R 2 teeth,

[0090] [ka]

[0091] and or R 2 teeth,

[0092] [ka]

[0093] and or R 2 teeth,

[0094] [ka]

[0095] and or R 2 teeth,

[0096] [ka]

[0097] (That is).

[0098] In one embodiment, the compound of formula (I) is n is 1 and R 1 is F and R 2 but

[0099] [ka]

[0100] (wherein X is N and R 5 is defined as CH3 or H, preferably CH3).

[0101] In one embodiment, the compound of formula (I) is n is 1 and R 1 is H and R 2 but

[0102] [ka]

[0103] (wherein X is N and R 5 is defined as CH3 or H, preferably CH3).

[0104] In one preferred embodiment, compounds of formula (I) are those in which n is 1 and R 1 is F or H, and R 2 but

[0105] [ka]

[0106] and or R 2 but,

[0107] [ka]

[0108] It is defined as:

[0109] In one embodiment, the compound of formula (I) is 1 is H and R 2 but

[0110] [ka]

[0111] (In the formula, R 3 is F and R 4 is -NH2 and R 7 is H and R 8 is H) It is defined as:

[0112] In one embodiment, the compound of formula (I) is 1 is F and R 2 but

[0113] [ka]

[0114] (In the formula, R 3 is F and R 4 is -NH2 and R7 is H and R 8 is H) It is defined as:

[0115] In one embodiment, the compound of formula (I) is 1 is F and R 2 but

[0116] [ka]

[0117] (In the formula, R 3 is H and R 4 is NH2 and R 7 is H and R 8 is CN) It is defined as:

[0118] In one embodiment, the compound of formula (I) is 1 is H or F, and R 2 but

[0119] [ka]

[0120] (In the formula, R 9 is selected from H, F, NH2 or CH3 In a preferred embodiment, R 9 is F. In a more preferred embodiment, R 9 is H.

[0121] In one embodiment, the compound of formula (I) is 1 is H or F, and R 2 but

[0122] [ka]

[0123] It is defined as:

[0124] In another embodiment, the compound of formula (I) is 1 is F and R 2 but,

[0125] [ka]

[0126] It is defined as:

[0127] In another embodiment, the compound of formula (I) is 1 is F and R 2 but,

[0128] [ka]

[0129] It is defined as:

[0130] In one embodiment, the compound of formula (I) is 1 is H or F, and R 2 but

[0131] [ka]

[0132] It is defined as:

[0133] In one embodiment, the compound of formula (I) is 1 is H or F, and R 2 but

[0134] [ka]

[0135] It is defined as:

[0136] In one embodiment, the compound of formula (I) is 1 is H or F, and R 2 but

[0137] [ka]

[0138] It is defined as:

[0139] In one embodiment, the compound of formula (I) is 1 is F and R 2 but,

[0140] [ka]

[0141] It is defined as:

[0142] In one embodiment, the compound of formula (I) is 1 is hydroxy(C1-C4)alkyl, preferably hydroxymethyl or hydroxyethyl, more preferably hydroxyethyl, and R 2 but

[0143] [ka]

[0144] (In the formula, R 9 is selected from H, F, NH2 or CH3 In a preferred embodiment, R 9 is H.

[0145] Preferred compounds of formula (I) are

[0146] [ka]

[0147] Includes.

[0148] In one embodiment, preferred compounds of formula (I) are the following stereoisomers:

[0149] [ka]

[0150] [ka]

[0151] may be selected from:

[0152] In one embodiment, the present invention relates to a compound of formula (I) comprising a detectable label. Preferably, the compound of formula (I) comprises one or more detectable labels.

[0153] The type of detectable label is not specifically limited and depends on the detection method selected.Examples of possible detectable labels include isotopes, such as radioisotopes (i.e., radionuclides), particularly positron emitters or gamma emitters.Detectable labels, such as radioisotopes, particularly positron emitters or gamma emitters, should be present in an amount that is not the same as the natural amount of each isotope.Furthermore, the amount used should allow its detection by the selected detection method.

[0154] In a preferred embodiment, the detectable label is 3 H and 18 F, most preferably 18 The detectable label may be present at any available position. Typically, the detectable label is a radioisotope of one of the atoms present in the compound of formula (I). For example, any reference to "F" in the present invention is 19 F (stable) or 18 Covers F (detectable label). Any reference to "H" 1H (stable) or 3 H (covers the detectable label, also called tritium, also designated "T" herein).

[0155] Isotopic variations of the compounds of the present invention can generally be prepared by conventional procedures, for example, by exemplary methods or by the preparations described in the following examples and preparations, using appropriate isotopic variations of suitable reagents that are commercially available or prepared by known synthetic techniques.Radioactive isotopes, particularly positron emitters or gamma emitters, can be included in the compounds of the present invention by methods conventional in the field of organic synthesis.Typically, they are introduced by using correspondingly labeled starting materials.Exemplary methods for introducing detectable labels are described, for example, in US 8,932,557, which is incorporated herein by reference.

[0156] 18 F may be attached at any location suitable for attaching fluorine. 18 F-labeled compounds are particularly suitable for imaging applications, such as positron emission tomography (PET). 19 The corresponding compounds containing F are also 18 It is of particular interest because it can be used as an analytical standard and reference during the manufacture, quality control, release and clinical use of the F-analogues.

[0157] In the compounds having formula (I), 18 F is, for example, R 2 as an F substituent in, or R 1 Preferably, this can be present as R 1 It can exist as.

[0158] 3 When H is used as the detectable label, it is preferably -CT3 (T is 3 H) and attached at any position where a CH3 group can be attached. Radioisotopes, e.g. 3Substitution with H offers certain diagnostic advantages resulting from greater metabolic stability, for example, by reducing defluorination, increasing in vivo half-life, or easing dosage requirements, while retaining or improving the efficacy of the original compound.

[0159] In one embodiment, the present invention provides a method for producing a fluorine-containing ... 3 Tritium (H) is replaced by a detectable label selected from 3 H), wherein one to three hydrogens (H) are replaced by tritium ( 3 H), which has been replaced by tritium ( 3 Preferably, compounds having formula (I) detectably labeled with tritium (H) are defined as compounds having formula (I) detectably labeled with tritium (H). 3 H), which has been replaced by tritium ( 3 More preferably defined are compounds having formula (I) detectably labeled with three hydrogens (H). 3 H), which has been replaced by tritium ( 3 Even more preferably defined are compounds having formula (I) detectably labeled with H).

[0160] In one embodiment, the present invention provides a compound of formula (I) having the formula (IT):

[0161] [ka]

[0162] (In the formula, n, R 1 , and R 2 is as defined herein for compounds of formula (I), and R 6 is T or H, and / or R 2 is substituted by at least one CT3, or R 2at least one of the hydrogen atoms in is replaced by T, and / or R 1 at least one hydrogen atom in 3 H) or a stereoisomer, polymorph, racemic mixture, tautomer, pharmaceutically acceptable salt, prodrug, hydrate or solvate thereof, or a mixture thereof. 6 is T or H, and / or R 2 is substituted by at least one CT3, or R 2 At least one of the hydrogen atoms in is replaced by T. T is 3 It's H.

[0163] In one embodiment, R 6 is T. In another embodiment, R 2 is substituted with at least one CT3. 2 At least one of the hydrogen atoms in R is replaced by T. In a preferred embodiment, R 6 is T and R 2 At least one of the hydrogen atoms in R is replaced by T. In yet another embodiment, R 1 At least one of the hydrogen atoms in is replaced by T.

[0164] In a preferred embodiment, the present invention provides a compound of formula (IT) (In the formula, n is 1 or 2, preferably n is 1; R 1 is H or F, R 6 is T or H, T is 3 H, R 2 teeth

[0165] [ka]

[0166] (In the formula, R 3 is F and R 4 is -NH2 and R 7 and R 8 at least one of is T and, if applicable, the other is H; Preferably R 7 is T and R 8 is T), or R 2 teeth

[0167] [ka]

[0168] (In the formula, R 8 is CN and R 4 is -NH2 and R 7 and R 3 is T and, if applicable, the other is H, and in one embodiment, R 7 is T and R 3 is T, and in another embodiment, R 7 is T and R 3 is H), R 2 teeth

[0169] [ka]

[0170] (wherein X is N and R 5 is tritiated CH3(CT3)), or R 2 teeth

[0171] [ka]

[0172] or R 2 teeth

[0173] [ka]

[0174] or R 2 teeth,

[0175] [ka]

[0176] (In the formula, R 9 is selected from H, F, NH2 or CH3

[0177] [ka]

[0178] (iii) selected from: R 2 teeth,

[0179] [ka]

[0180] (In the formula, R 9 is selected from H, F, NH2 or CH3, preferably NH2 or F or R 2 teeth,

[0181] [ka]

[0182] The present invention relates to a compound of formula (I)

[0183] In one embodiment, the present invention provides a compound of formula (IT):

[0184] [ka]

[0185] (In the formula, n is 1 or 2, R 1 is H, hydroxy(C1-C4)alkyl or F, T is 3 H, R 2 teeth

[0186] [ka]

[0187] (In the formula, R 3 is F and R 4 is NH2 and R 7 and R 8 at least one of which is T and, if applicable, the other is H, preferably R 7 is T and R 8 is T), and R 6 is T Or, R 2 teeth

[0188] [ka]

[0189] (In the formula, R 4 is NH2 and R 8 is CN and R 3 or R 7 at least one of which is T and, if applicable, the other is H, preferably R 7 is T and R 3 is T, or R 7 is T and R 3 is H), R 6 is T, Or, R2 teeth

[0190] [ka]

[0191] (wherein X is N and R 5 is CT3), R 6 is H, or R 2 teeth

[0192] [ka]

[0193] (In the formula, R 12 is T), or R 2 teeth

[0194] [ka]

[0195] (In the formula, R 12 is T), Or, R 2 teeth

[0196] [ka]

[0197] (In the formula, R 12 is T), Or, R 2 teeth

[0198] [ka]

[0199] (In the formula, R 12is T) or a stereoisomer, polymorph, racemic mixture, tautomer, pharmaceutically acceptable salt, prodrug, hydrate or solvate thereof.

[0200] In another embodiment, the present invention provides a compound of formula (IT) (In the formula, n is 1 or 2, preferably n is 2; R 1 is H or F (preferably H), R 6 is T, R 2 teeth

[0201] [ka]

[0202] (In the formula, R 3 is F and R 4 is NH2 and R 7 and R 8 wherein at least one of is T and, if applicable, the other is H).

[0203] Preferably R 7 is T and R 8 is T, Or, R 2 teeth

[0204] [ka]

[0205] (In the formula, R 8 is CN and R 4 is NH2 and R 7 and R 3 where at least one of R is T and, if applicable, the other is H; 7 is T and R 3 is T, or in another embodiment, R 7is T and R 3 is H, R 2 teeth

[0206] [ka]

[0207] (wherein X is N and R 5 is tritiated CH3(CT3)), Or, R 2 teeth

[0208] [ka]

[0209] or R 2 teeth

[0210] [ka]

[0211] or R 2 teeth,

[0212] [ka]

[0213] (In the formula, R 9 is selected from H, F, NH2 or CH3

[0214] [ka]

[0215] from,

[0216] [ka]

[0217] (iii) selected from the group consisting of: 2 teeth,

[0218] [ka]

[0219] (In the formula, R 9 is selected from H, F, NH2 or CH3, preferably NH2 or F, or R 2 teeth,

[0220] [ka]

[0221] In one embodiment, the present invention provides a compound of formula (IT) (In the formula, n is 1 or 2, preferably n is 2; R 1 is F, R 6 is H, R 2 teeth

[0222] [ka]

[0223] (wherein X is N and R 5 is tritiated CH3(CT3))

[0224] In one embodiment, the present invention provides a compound of formula (IT) (In the formula, n is 1 or 2, preferably n is 2; R 1 is H or F, R 6 is T, R2 teeth

[0225] [ka]

[0226] (In the formula, R 12 relates to compounds in which T is

[0227] In one embodiment, the present invention provides a compound of formula (IT) (In the formula, n is 1 or 2, preferably n is 2; R 1 is H or F, R 6 is T, R 2 teeth

[0228] [ka]

[0229] The present invention relates to a compound of formula (I)

[0230] In one embodiment, the present invention provides a compound of formula (IT) (In the formula, n is 1 or 2, preferably n is 2; R 1 is H or F, R 6 is T, R 2 teeth

[0231] [ka]

[0232] The present invention relates to a compound of formula (I)

[0233] In another embodiment, the present invention provides a compound of formula (IT) (In the formula, n is 1 or 2, R1 is H or F, preferably H, R 6 is T, R 2 teeth

[0234] [ka]

[0235] (In the formula, R 9 is selected from H, F, NH or CH, preferably NH or F, or R 2 teeth,

[0236] [ka]

[0237] The present invention relates to a compound of formula (I)

[0238] In a most preferred embodiment, the present invention provides a compound of formula (IT) (In the formula, n is 1 or 2, preferably n is 2; R 1 is hydroxy(C1-C4)alkyl, preferably hydroxyethyl or hydroxymethyl, more preferably hydroxymethyl; R 1 At least one hydrogen atom in is replaced by T, preferably R 1 Two of the hydrogen atoms in are replaced by T, R 6 is T or H, T is 3 H, R 2 is a 5- or 6-membered carbocyclic or heterocyclic ring optionally substituted with F, NH, CN and / or CH, the heterocyclic ring containing one or more heteroatoms selected from N, O and S, preferably N.

[0239] Preferred tritium ( 3 H) the detectably labeled compound is

[0240] [ka]

[0241] (Wherein, T is 3 (meaning H) In a preferred embodiment, the tritium ( 3 H) The detectably labeled compound is a stereoisomer.

[0242] [ka]

[0243] (Wherein, T is 3 (meaning H) In a more preferred embodiment, the tritium ( 3 H) The detectably labeled compound is a stereoisomer.

[0244] [ka]

[0245] (Wherein, T is 3 (meaning H) It could be.

[0246] In another more preferred embodiment, the tritium ( 3 H) the detectably labeled compound is

[0247] [ka]

[0248] (Wherein, T is 3 (meaning H) It could be.

[0249] In one embodiment, the present invention provides a compound of formula (IF)

[0250] [ka]

[0251] (In the formula, n, R 1 and R 2 is as defined herein for compounds of formula (I), and at least one F is 18 F) 18 Provided is a compound detectably labeled with F, or a stereoisomer, polymorph, racemic mixture, tautomer, pharmaceutically acceptable salt, prodrug, hydrate or solvate thereof, or a mixture thereof.

[0252] In a preferred embodiment, the present invention relates to compounds of formula (IF), wherein R 1 teeth, 18 F (detectable label).

[0253] In a further preferred embodiment, n is 1 or 2, R 1 teeth, 18 F (detectable label), R 2 teeth,

[0254] [ka]

[0255] (wherein X is N and R 5 is CH3 or H, preferably CH3), or R 2 teeth,

[0256] [ka]

[0257] or R 2 teeth,

[0258] [ka]

[0259] or R 2 teeth,

[0260] [ka]

[0261] (In the formula, R 3 is F and R 4 is NH2 and R 7 is H and R 8 is H, or R 3 is H and R 4 is NH2 and R 7 is H and R 8 is CN), or R 2 teeth,

[0262] [ka]

[0263] (In the formula, R 9 is selected from H and F, NH2 or CH3, or R 2 teeth,

[0264] [ka]

[0265] or R 2 teeth,

[0266] [ka]

[0267] or R 2 teeth,

[0268] [ka]

[0269] or R 2 teeth,

[0270] [ka]

[0271] or R 2 teeth,

[0272] [ka]

[0273] or R 2 teeth,

[0274] [ka]

[0275] is.

[0276] In a preferred embodiment, the present invention provides a compound of formula (IF) (In the formula, n is 1 or 2, R 1 teeth, 18 F (detectable label), R 2 teeth,

[0277] [ka]

[0278] (In the formula, R 9 is H).

[0279] In a preferred embodiment, n is 1. In another preferred embodiment, n is 2.

[0280] In another embodiment, (IF) is

[0281] [ka]

[0282] (In the formula, n, R 1 and R 2 is as defined herein for compounds of formula (I) or a stereoisomer, polymorph, racemic mixture, tautomer, pharmaceutically acceptable salt, prodrug, hydrate or solvate thereof, or a mixture thereof.

[0283] In another preferred embodiment, (IF) is the following compound:

[0284] [ka]

[0285] (wherein n is 1 or 2, R 1 is hydroxy(C1-C4)alkyl, preferably hydroxyethyl or hydroxymethyl, more preferably hydroxymethyl; R 2 is a 5- or 6-membered carbocyclic or heterocyclic ring optionally substituted with F, NH, CN and / or CH, the heterocyclic ring containing one or more heteroatoms selected from N, O and S, preferably N).

[0286] In a preferred embodiment, R 2 teeth

[0287] [ka]

[0288] (wherein X is N and R 5 is CH3 or H, preferably CH3), Or, R 2 teeth

[0289] [ka]

[0290] (In the formula, R 3 is F and R 4 is NH2 and R 7 is H and R 8 is H, or R 3 is H and R 4 is NH2 and R 7 is H and R 8 is CN), or R 2 teeth,

[0291] [ka]

[0292] (In the formula, R 9 is selected from H and F, NH2 or CH3, or R 2 teeth,

[0293] [ka]

[0294] or R 2 teeth,

[0295] [ka]

[0296] or R 2 teeth,

[0297] [ka]

[0298] or R 2 teeth,

[0299] [ka]

[0300] or R 2 teeth,

[0301] [ka]

[0302] or R 2 teeth,

[0303] [ka]

[0304] is.

[0305] In one preferred embodiment, n is 1 or 2, preferably 2; R 1 is hydroxy(C1-C4)alkyl, preferably hydroxyethyl or hydroxymethyl, more preferably hydroxymethyl; R 2 teeth,

[0306] [ka]

[0307] (In the formula, R 9 is selected from H and F, NH2 or CH3, preferably H).

[0308] In some embodiments, n is 1. In preferred embodiments, n is 2.

[0309] In one preferred embodiment, the present invention provides a compound of formula (IF)

[0310] [ka]

[0311] (In the formula, n is 1 or 2, preferably 2; R 1 is hydroxy(C1-C4)alkyl, preferably hydroxyethyl or hydroxymethyl, more preferably hydroxymethyl; R 2 teeth,

[0312] [ka]

[0313] (In the formula, R 9 is H).

[0314] Preferred compounds of formula (IF) according to the present invention 18 A compound detectably labeled with F is

[0315] [ka]

[0316] may be selected from:

[0317] More preferably, the compound of formula (IF) according to the present invention 18 Compounds detectably labeled with F may be used to identify stereoisomers.

[0318] [ka]

[0319] It could be.

[0320] Other preferred compounds of formula (IF) according to the present invention 18 A compound detectably labeled with F is

[0321] [ka]

[0322] It could be.

[0323] Diagnostic Compositions In a second aspect, the present invention relates to a diagnostic composition comprising a compound of formula (I) as defined above and, optionally, at least one physiologically acceptable carrier, diluent, adjuvant and / or excipient.

[0324] The compounds of the present invention are particularly suitable for imaging TDP-43 aggregates. The imaging can be performed in mammals, preferably humans. The imaging is preferably in vitro imaging, ex vivo imaging, or in vivo imaging. More preferably, the imaging is in vivo imaging. Even more preferably, the imaging is brain imaging. The imaging can also be ocular / retinal imaging or imaging of tissues of the central nervous system.

[0325] The compounds of the present invention are particularly suitable for use in diagnostic methods.The diagnostic method can be carried out on mammals, preferably humans.The target tissue on which the diagnostic method is carried out can be brain tissue, central nervous system tissue, eye tissue (for example, retinal tissue) or other tissue, or body fluid, such as cerebrospinal fluid (CSF).The tissue is preferably brain tissue.

[0326] A "diagnostic composition" is defined herein as a composition comprising one or more compounds of the invention in a form suitable for administration to a patient (e.g., a mammal, e.g., a human) and for use in diagnosing a particular disease, disorder, or abnormality of interest. In one embodiment, the diagnostic composition comprises a detectably labeled compound of the invention as described above, and optionally at least one physiologically acceptable carrier, diluent, adjuvant, and / or excipient.

[0327] Preferred detectably labeled compounds of the invention are of formula (IT) or (IF):

[0328] The diagnostic composition is suitable for use in diagnosing a disease, disorder, or disorder associated with TDP-43 aggregates, or a TDP-43 proteinopathy, as defined herein below. Preferably, the diagnostic composition optionally further comprises a physiologically acceptable excipient, carrier, diluent, or adjuvant. Administration is preferably carried out as defined below. More preferably, by injection of the composition as an aqueous solution. The diagnostic composition may optionally contain additional components, such as a buffer; a pharmaceutically acceptable solubilizer (e.g., cyclodextrin or a surfactant, such as Pluronic, Tween, or phospholipid); and a pharmaceutically acceptable stabilizer or antioxidant (e.g., ascorbic acid, gentisic acid, or para-aminobenzoic acid). Dosages of the compounds of the present invention will vary depending on the exact compound administered, the patient's weight, and other variables apparent to a practitioner skilled in the art.

[0329] While the compounds of the present invention can be administered alone, they are preferably formulated into diagnostic compositions according to standard pharmaceutical practice. Thus, diagnostic compositions comprising a diagnostically effective amount of a compound of the present invention in combination with a pharmaceutically acceptable carrier, diluent, adjuvant, and / or excipient are part of the present invention. Preferred pharmaceutically acceptable carriers, diluents, adjuvants, and / or excipients are those that are physiologically compatible with the diagnostic compositions according to the present invention.

[0330] Pharmaceutically acceptable excipients are well known in the pharmaceutical industry and are described, for example, in Remington's Pharmaceutical Sciences, 18th Edition (Alfonso R. Gennaro, ed.; Mack Publishing Company, Easton, PA, 1990). Pharmaceutically acceptable excipients can be selected with regard to the intended route of administration and standard pharmaceutical practice. An excipient must be acceptable in the sense of not being harmful to the recipient thereof.

[0331] Pharmaceutically useful excipients, carriers, adjuvants and diluents that can be used in formulating the diagnostic compositions of the present invention may include, for example, solvents, such as monohydric alcohols, e.g., ethanol, isopropanol and polyhydric alcohols, e.g., glycols and edible oils, e.g., soybean oil, coconut oil, olive oil, safflower oil, cottonseed oil, oily esters, e.g., ethyl oleate, isopropyl myristate, binders, adjuvants, solubilizers, thickeners, stabilizers, disintegrants, glidants, lubricants, buffers, emulsifiers, wetting agents, suspending agents, sweeteners, colorants, flavorings, coating agents, preservatives, antioxidants, processing agents, drug delivery modifiers and enhancers, e.g., calcium phosphate, magnesium stearate, talc, monosaccharides, disaccharides, starch, gelatin, cellulose, methylcellulose, sodium carboxymethylcellulose, dextrose, hydroxypropyl-β-cyclodextrin, polyvinylpyrrolidone, low melting point waxes and ion exchange resins.

[0332] The route for administering (delivering) the compound of the present invention includes, but is not limited to, one or more of intravenous, gastrointestinal, intrathecal, intraperitoneal, intramuscular, oral (e.g., as a tablet, capsule, or ingestible liquid), topical, mucosal (e.g., as a nasal spray or aerosol for inhalation), intranasal, parenteral (e.g., by an injectable form), intrauterine, intraocular, intradermal, intracranial, intratracheal, intravaginal, intraventricular, intracerebral, subcutaneous, intraocular (including intravitreal or intracameral), transdermal, rectal, buccal, epidural, and sublingual. Preferably, the route for administering (delivering) the compound of the present invention is parenteral.

[0333] The compounds of the invention (e.g., detectably labeled compounds, e.g., 3 H or 18 When a compound (having a detectable label with F) is administered parenterally, examples of such routes of administration include one or more of intravenous, intraarterial, intraperitoneal, intrathecal, intraventricular, intraurethral, ​​intrasternal, intracranial, intramuscular, or subcutaneous, and / or by use of injection techniques. For parenteral administration, the compound is best used in the form of a sterile aqueous solution which may contain other excipients. The aqueous solution should be suitably buffered (preferably to a pH of from 3 to 9), if necessary. The preparation of suitable parenteral formulations under sterile conditions is readily accomplished by standard pharmaceutical techniques well known to those skilled in the art.

[0334] Typically, a physician will determine the actual dosage that will be most suitable for an individual patient. The dosage of a compound of the invention (e.g., a detectably labeled compound, e.g., 3 H or 18 The dose of the radiation (having a detectable label with F) will vary depending on the exact compound administered, the patient's weight, the size and type of sample, and other variables apparent to a practitioner of ordinary skill in the art. Generally, the dose can range preferably from 0.001 μg / kg to 10 μg / kg, preferably from 0.01 μg / kg to 1.0 μg / kg. The radiation dose can be, for example, from 100 to 600 MBq, more preferably from 150 to 450 MBq.

[0335] Due to their design and binding properties, the compounds of the invention as defined herein can be used in the diagnosis of diseases, disorders and conditions associated with TDP-43 aggregates. The compounds of the invention are particularly suitable for positron emission tomography imaging of TDP-43 aggregates.

[0336] The compounds of the invention disclosed herein may be used to treat diseases, disorders or conditions associated with TDP-43 aggregates or TDP-43 proteinopathies, such as frontotemporal dementia (FTD, e.g., sporadic or familial corticobasal degeneration, frontotemporal lobar dementia TDP-43, with or without motor neuron disease (MND), with progranulin (GRN) mutations, with C9orf72 mutations, with TARDBP mutations, with valosin-containing protein (VCP) mutations, linked to chromosome 9p, Frontotemporal lobar degeneration (FTLD) with TDP-43 (including TDP-43) or frontotemporal lobar degeneration with ubiquitin-positive TDP-43 inclusions (FTLD-TDP), argyrophilic grain dementia, Pick's disease, semantic primary progressive aphasia (svPPA), behavioral FTD (bvFTD), non-fluent primary progressive aphasia (e.g., nfvPPA), amyotrophic lateral sclerosis (ALS, e.g., with TARDBP mutations, with angiogenin (ANG) mutations) Sporadic ALS), Alexander disease (AxD), limbic-predominant age-related TDP-43 encephalopathy (LATE), chronic traumatic encephalopathy, Perry syndrome, Alzheimer's disease (including AD, sporadic and familial forms of AD), Down syndrome, familial British dementia, polyglutamine diseases (Huntington's disease and spinocerebellar ataxia type 3 (SCA3; also known as Machado-Joseph disease)), hippocampal sclerosis dementia and myopathy (sporadic inclusion body myositis, Baro and inclusion body myopathy associated with mutations in the myotilin-containing protein (VCP); also Paget's disease of bone and frontotemporal dementia), oculopharyngeal muscular dystrophy with rimmed vacuoles, myofibrillar myopathy associated with mutations in the myotilin (MYOT) gene or mutations in the gene encoding desmin (DES), traumatic brain injury (TBI), dementia with Lewy bodies (DLB), and Parkinson's disease (PD), preferably, the TDP-43 aggregate-associated disease, disorder, or abnormality is selected from frontotemporal dementia (FTD), amyotrophic lateral sclerosis (ALS), Alzheimer's disease (AD), Parkinson's disease (PD), chronic traumatic encephalopathy (CTE), and limbic-predominant age-related TDP-43 encephalopathy (LATE).

[0337] In one embodiment, the disease, disorder or condition associated with TDP-43 aggregates or TDP-43 proteinopathy is amyotrophic lateral sclerosis (ALS).

[0338] In one embodiment, the diagnosis of a disease, disorder or condition associated with TDP-43 aggregates or a TDP-43 proteinopathy is Alzheimer's disease (AD).

[0339] In one embodiment, the diagnosis of a disease, disorder, or abnormality associated with TDP-43 aggregates or a TDP-43 proteinopathy is frontotemporal dementia (FTD), including frontotemporal lobe dementia associated with TDP-43, or frontotemporal lobar degeneration with TDP-43 inclusions (FTLD-TDP).

[0340] In one embodiment, the diagnosis of a disease, disorder or abnormality associated with TDP-43 aggregates or a TDP-43 proteinopathy is limbic-predominant age-related TDP-43 encephalopathy (LATE).

[0341] Methods and Uses In a third aspect, the present invention provides the methods and uses listed below. A method for imaging a disease, disorder, or abnormality associated with TDP-43 aggregates in a subject; A method for positron emission tomography (PET) imaging of TDP-43 aggregates in tissue of a subject, A method for detecting, and optionally quantifying, TDP-43 aggregates in tissue of a subject; A method for diagnostic imaging of the brain of a subject; A method for determining the amount of TDP-43 aggregates in a sample or in a specific body part or body region; a method for diagnosing a disease, disorder, or abnormality associated with TDP-43 aggregates or for diagnosing a TDP-43 proteinopathy; A method for collecting data for diagnosing a disease, disorder, or abnormality associated with TDP-43 aggregates or for diagnosing a TDP-43 proteinopathy; A method for collecting data to determine a disease, disorder, or condition associated with TDP-43 aggregates, or a predisposition to a TDP-43 proteinopathy; a method for collecting data for monitoring the progression of a disease, disorder or condition associated with TDP-43 aggregates or for monitoring the progression of a TDP-43 proteinopathy in a patient; A method for collecting data to predict the responsiveness of a patient suffering from a disease, disorder, or condition associated with TDP-43 aggregates to treatment with a pharmaceutical agent; Use of the compounds of the present invention as biomarkers for TDP-43 aggregates or as biomarkers for TDP-43 proteinopathy. Use of the compounds of the present invention as diagnostic agents or tools for TDP-43 proteinopathy. Use of the compounds of the invention as in vitro analytical standards or in vitro screening tools Regarding.

[0342] Any of the compounds of the present invention (e.g., compounds of formula (I), (IT), or (IF)) can be used in the methods summarized above. Preferably, the compound is a detectably labeled compound (e.g., 3 H or 18 F) with a detectable label.

[0343] The methods of the invention may include contacting a sample, a particular body part, or a body region suspected of containing TDP-43 aggregates with a compound of the invention.

[0344] The body is preferably that of a mammal, more preferably a human, which includes the whole body or partial body regions / parts of a patient suspected of containing TDP-43 aggregates.

[0345] The sample may be selected from tissues or body fluids suspected of containing TDP-43 aggregates, and the sample is obtained from a patient. Preferably, the tissue is selected from tissues of the central nervous system (CNS), eye tissue, or brain tissue, more preferably brain tissue. Examples of body fluids include cerebrospinal fluid (CSF) or blood. The sample is obtained from a mammal, more preferably a human. Preferably, the sample is an in vitro sample from a patient.

[0346] In vitro samples obtained from a patient, or specific body parts or regions, can be contacted with the compounds of the invention by direct incubation.

[0347] In in vivo methods, a specific body part or region can be contacted with a compound of the invention by administering an effective amount of the compound to a patient, the effective amount being an amount that allows the presence or absence of TDP-43 aggregates to be determined in the specific body part or region using a selected analytical technique.

[0348] The step of binding the compounds of the present invention to TDP-43 aggregates includes allowing a sufficient amount of time for the binding to occur. The amount of time required for binding depends on the type of test (e.g., in vitro or in vivo) and can be determined by one of ordinary skill in the art through routine experimentation. For in vitro methods, the amount of time depends on the sample or the specific body part or region, and can range, for example, from about 30 minutes to about 120 minutes. For in vivo methods, the amount of time depends on the time required for the compounds of the present invention to reach the specific body part or region suspected of containing TDP-43 aggregates. The amount of time should not be too long to avoid washout and / or metabolism of the compounds of the present invention. The duration can range, for example, from about 0 minutes to about 240 minutes (this is the duration of the PET scan during initial compound characterization (NHP PET and subsequent FiH studies)).

[0349] The method for detecting the compound of the present invention bound to TDP-43 aggregates is not particularly limited and depends, inter alia, on the type of detectable label, the sample, the specific body part or region, and whether the method is an in vitro or in vivo method. Possible detection methods include, but are not limited to, fluorescent or nuclear imaging techniques, such as positron emission tomography (PET), single-photon emission computed tomography (SPECT), magnetic resonance imaging (MRI), and contrast-enhanced magnetic resonance imaging (MRI). Fluorescent and / or nuclear imaging techniques can be used to monitor and / or visualize the distribution of the compound of the present invention within a sample or body. The imaging system is designed to provide an image of the detectably bound label, such as a radioisotope, particularly a positron emitter or gamma emitter, present in the tested sample, specific body part, or tested body region. Preferably, the compound of the present invention bound to TDP-43 aggregates is detected by an imaging device, such as a PET or SPECT scanner. The amount of compound bound to TDP-43 can be determined by visual or quantitative analysis using, for example, a PET scan image.

[0350] In one embodiment, the presence or absence of a compound of the present invention bound to TDP-43 aggregates can be correlated with the presence or absence of TDP-43 aggregates in a sample or in a particular body part or region. The correlation can be qualitative or quantitative. In a preferred embodiment, this step comprises: - determining the amount of the compound of the invention that is bound to the TDP-43 aggregates; - correlating the amount of the compound of the invention bound to TDP-43 aggregates with the amount of TDP-43 aggregates in the sample or in a particular body part or body region; and - optionally comparing the amount of compound bound to TDP-43 aggregates in the sample or in a particular body part or body region with a normal control value in a healthy control subject. Includes.

[0351] The amount of compound bound to TDP-43 aggregates can be determined by any suitable method, a preferred method being positron emission tomography (PET).

[0352] In another embodiment, the presence or absence of a compound of the present invention bound to TDP-43 aggregates can be correlated with a disease, disorder, or condition associated with TDP-43 aggregates, or a TDP-43 proteinopathy, or a predisposition thereto. The correlation can be qualitative or quantitative. In a preferred embodiment, this step comprises: - determining the amount of the compound of the invention that is bound to the TDP-43 aggregates; - correlating the amount of the compound of the invention bound to TDP-43 aggregates with the amount of TDP-43 aggregates in the sample or in a particular body part or body region; and - optionally comparing the amount of compound bound to TDP-43 aggregates in the sample or in a particular body part or body region with a normal control value in a healthy control subject. Includes.

[0353] In any of the methods disclosed herein, steps (a) to (c) and, if present, optional step (d) may be repeated at least once. Repeating the steps is particularly useful for methods that collect data for monitoring progression and for predicting responsiveness. In these methods, it may be advisable to monitor the patient over time and repeat the above steps after a certain period of time has passed. The time interval before repeating the above-mentioned steps can be determined by a physician depending on the severity of the disease, disorder, or abnormality associated with TDP-43 aggregates or TDP-43 proteinopathy.

[0354] In one embodiment, the present invention provides a method for detecting a neurological disease, disorder, or abnormality associated with, or a predisposition thereto, associated with TDP-43 aggregates in a subject, comprising: (a) administering to a subject a compound of the invention disclosed herein or a diagnostic composition comprising a compound of the invention. (b) binding the compound to TDP-43 aggregates; and (c) detecting a compound bound to TDP-43 aggregates The present invention relates to a method comprising:

[0355] In one embodiment, the present invention provides a method (e.g., an in vivo or in vitro method) for detecting, and optionally quantitating, TDP-43 aggregates in a tissue of a subject, comprising: (a) administering to a subject a compound of the invention disclosed herein or a diagnostic composition comprising a compound of the invention disclosed herein; (b) binding the compound to TDP-43 aggregates; and (c) detecting, and optionally quantifying, compounds bound to TDP-43 aggregates using positron emission tomography. The present invention relates to a method comprising:

[0356] In one embodiment, the present invention provides a method of diagnostic imaging of the brain of a subject, comprising: (a) administering to a subject a compound of the invention disclosed herein or a diagnostic composition comprising a compound of the invention. (b) binding the compound to TDP-43 aggregates; and (c) detecting the compound bound to the TDP-43 aggregates by collecting positron emission tomography (PET) images of the subject's brain. The present invention relates to a method comprising:

[0357] Imaging: The present invention relates to a method for imaging TDP-43 aggregates using the compounds of the present invention. Imaging can be performed, for example, using any of the methods mentioned above, in particular by PET.

[0358] In one embodiment, the present invention provides a method for imaging TDP-43 aggregates in a sample or a patient, particularly in the brain or a sample obtained from the brain of a patient, comprising the steps of: (a) administering to a subject a compound of the invention disclosed herein or a diagnostic composition comprising a compound of the invention. (b) binding the compound to TDP-43 aggregates; and (c) detecting a compound bound to TDP-43 aggregates The present invention relates to a method comprising:

[0359] In one embodiment, the present invention provides a method of imaging or diagnosing a disease, disorder, or condition associated with, or a predisposition to, TDP-43 aggregates in a subject, comprising: (a) administering to a subject a compound of the invention disclosed herein or a diagnostic composition comprising a compound of the invention. (b) binding the compound to TDP-43 aggregates; and (c) detecting the compound binding to TDP-43 aggregates in the brain of the subject. The present invention relates to a method comprising:

[0360] In one embodiment, the present invention provides a method of imaging or diagnosing a disease, disorder, or condition associated with, or a predisposition to, TDP-43 aggregates in a subject, comprising: (a) administering to a subject a compound of the invention disclosed herein or a diagnostic composition comprising a compound of the invention. (b) binding the compound to TDP-43 aggregates; and (c) detecting a compound bound to TDP-43 aggregates The present invention relates to a method comprising:

[0361] In one embodiment, the present invention provides a method of imaging or diagnosing a disease, disorder, or condition associated with, or a predisposition to, TDP-43 aggregates in a subject, comprising: (a) administering to a subject a compound of the invention disclosed herein or a diagnostic composition comprising a compound of the invention. (b) binding the compound to TDP-43 aggregates; (c) detecting compounds that bind to TDP-43 aggregates; and (d) generating an image showing the location and / or amount of the compound bound to the TDP-43 aggregates The present invention relates to a method comprising:

[0362] In one embodiment, the present invention provides a method for positron emission tomography (PET) imaging of TDP-43 aggregates in tissue of a subject, comprising: (a) administering to a subject a compound of the invention disclosed herein or a diagnostic composition comprising a compound of the invention. (b) binding the compound to TDP-43 aggregates; and (c) detecting the compound bound to the TDP-43 aggregates by collecting positron emission tomography (PET) images of the subject's tissue. The present invention relates to a method comprising:

[0363] Preferably, the tissue is central nervous system (CNS) tissue, eye tissue or brain tissue, more preferably, the tissue is brain tissue.

[0364] In one embodiment, the present invention provides a method for imaging TDP-43 aggregates in a sample or a patient, comprising: (a) contacting a sample, a specific body part, or a body region suspected of containing TDP-43 aggregates with a compound of the invention disclosed herein, or a diagnostic composition comprising a compound of the invention; (b) binding the compound to TDP-43 aggregates; and (c) detecting compounds that bind to TDP-43 aggregates by imaging the sample, specific body parts, or body regions with an imaging system. The present invention relates to a method comprising:

[0365] In one embodiment, the present invention provides a method for imaging TDP-43 aggregates in an in vitro patient sample, comprising: (a) contacting an in vitro sample suspected of containing aggregated TDP-43 with a compound of the invention disclosed herein or a diagnostic composition comprising a compound of the invention; (b) binding the compound to TDP-43 aggregates; and (c) detecting the compound bound to the TDP-43 aggregates by imaging the in vitro sample with an imaging system. The present invention relates to a method comprising:

[0366] In one embodiment, the present invention provides a method for imaging TDP-43 aggregates in a patient, or in a particular body part or region of a patient, comprising: (a) contacting a sample, or a specific body part or region, suspected of containing TDP-43 aggregates with a compound of the present invention, preferably a compound of formula (IT) or formula (IF), or a diagnostic composition comprising a compound of the present invention disclosed herein, preferably a compound of formula (IT) or formula (IF); (b) binding the compound to TDP-43 aggregates; and (c) detecting compounds bound to TDP-43 aggregates by imaging the sample or a particular body part or region of the patient with an imaging system; The present invention relates to a method comprising:

[0367] The step of imaging the sample, patient, or specific body part or region of the patient with an imaging system includes detecting the compound of the present invention bound to TDP-43 aggregates using the imaging system disclosed herein. Detecting the compound of the present invention bound to TDP-43 aggregates allows imaging and identification of the distribution of TDP-43 aggregates in the tested sample, patient, specific body part, or region. PET imaging should be performed when the compound has penetrated the tissue and the compound is bound to the TDP-43 aggregates.

[0368] Determination of the amount of TDP-43 aggregates: In one embodiment, the present invention relates to a method for determining the amount of TDP-43 aggregates in a sample, specific body part, or body region suspected of containing TDP-43 aggregates using the compounds of the present invention.

[0369] In one embodiment, the present invention provides a method for determining the amount of TDP-43 aggregates in a sample, specific body part, or body region suspected of containing TDP-43 aggregates, comprising: (a) contacting a sample, a specific body part, or a body region suspected of containing TDP-43 aggregates with a compound of the invention disclosed herein, or a diagnostic composition comprising a compound of the invention; (b) binding the compound of the present invention to TDP-43 aggregates; (c) detecting the compound of the present invention bound to TDP-43 aggregates; (d) determining the amount of the compound of the present invention that is bound to the TDP-43 aggregates; and (e) optionally calculating the amount of TDP-43 aggregates in the sample, a particular body part, or a body region; The present invention provides a method comprising:

[0370] The radioactive signal is generated by at least one radiolabeled atom (e.g. 3 H, 2 H or 18 F) is observed when the detectably labeled compound of the invention binds to TDP-43 aggregates.

[0371] diagnosis: In one embodiment, the present invention provides a method for diagnosing a disease, disorder, or condition associated with TDP-43 aggregates, or a TDP-43 proteinopathy, or a predisposition thereto, comprising: (a) detecting a compound of the present invention bound to TDP-43 aggregates; and (b) correlating the presence or absence of the compound of the present invention bound to TDP-43 aggregates with a disease, disorder, or abnormality associated with TDP-43 aggregates, or a TDP-43 proteinopathy. The present invention relates to a method comprising:

[0372] Preferably, the method for diagnosing a disease, disorder or abnormality associated with TDP-43 aggregates, or a TDP-43 proteinopathy, or a predisposition thereto, comprises: (a) contacting a sample, a specific body part, or a body region suspected of containing TDP-43 aggregates with a compound of the invention disclosed herein, or a diagnostic composition comprising a compound of the invention; (b) binding the compound of the present invention to TDP-43 aggregates; (c) detecting the compound of the present invention bound to TDP-43 aggregates; and (d) correlating the presence or absence of the compound of the present invention bound to TDP-43 aggregates with a disease, disorder, or abnormality associated with TDP-43 aggregates, or a TDP-43 proteinopathy. Includes.

[0373] In one embodiment, the present invention provides a method for collecting data for diagnosing a disease, disorder or condition associated with TDP-43 aggregates, or a TDP-43 proteinopathy, or a predisposition thereto, comprising the steps of: (a) contacting a sample suspected of containing TDP-43 aggregates, or a specific body part or body region, with a compound of the invention disclosed herein or a diagnostic composition comprising a compound of the invention; (b) binding the compound of the present invention to TDP-43 aggregates; (c) detecting the compound of the present invention bound to TDP-43 aggregates; and (d) optionally correlating the presence or absence of a compound of the invention bound to TDP-43 aggregates with the presence or absence of TDP-43 aggregates in the sample or in a particular body part or body region. The present invention relates to a method comprising:

[0374] After contacting a sample or a specific body part or region with a compound of the present invention, the compound is allowed to bind to TDP-43 aggregates. The amount of time required for binding depends on the type of test (e.g., in vitro or in vivo) and can be determined by routine experimentation by one skilled in the art. Compounds bound to TDP-43 aggregates can then be detected by any suitable method. The specific method selected will depend on the detectable label selected. Examples of possible methods include, but are not limited to, fluorescent imaging or nuclear imaging techniques, such as positron emission tomography (PET), single-photon emission computed tomography (SPECT), magnetic resonance imaging (MRI), and contrast-enhanced magnetic resonance imaging (MRI). Fluorescent and / or nuclear imaging techniques can be used to monitor and / or visualize the distribution of detectably labeled compounds within a sample or within a specific body part or region.

[0375] Optionally, the step of correlating the presence or absence of a compound bound to TDP-43 aggregates with the presence or absence of TDP-43 aggregates in a sample or a particular body part or body region referred to herein above may comprise: - determining the amount of the compound bound to the TDP-43 aggregates; - correlating the amount of compound bound to TDP-43 aggregates with the amount of TDP-43 aggregates in the sample or in a particular body part or body region; - optionally comparing the amount of compound bound to TDP-43 aggregates in the sample or in a particular body part or body region with a normal control value in a healthy control subject. Includes.

[0376] The amount of compound bound to TDP-43 aggregates can be compared to a normal control value determined in a sample from a healthy subject or in a specific body part or region, and an increase in the amount of compound bound to TDP-43 aggregates compared to the normal control value can indicate that the patient is suffering from or at risk of developing a disease, disorder, or condition associated with TDP-43 aggregates.

[0377] If the amount of compound bound to TDP-43 aggregates is higher than the normal control value as defined herein, the patient can be predicted to be suffering from or susceptible to a disease, disorder or condition associated with TDP-43 aggregates or a TDP-43 proteinopathy.

[0378] Determining predisposing factors: A further aspect of the present invention relates to a method of collecting data for determining a predisposition to a disease, disorder or condition associated with TDP-43 aggregates or a TDP-43 proteinopathy, the method comprising: (a) contacting a sample suspected of containing TDP-43 aggregates, or a specific body part or body region, with a compound of the invention disclosed herein or a diagnostic composition comprising a compound of the invention; (b) binding the compound of the present invention to TDP-43 aggregates; (c) detecting the compound of the present invention bound to TDP-43 aggregates; and (d) optionally correlating the presence or absence of a compound of the invention bound to TDP-43 aggregates with the presence or absence of TDP-43 aggregates in the sample or in a particular body part or body region. Includes.

[0379] Optionally, the step of correlating the presence or absence of a compound bound to TDP-43 aggregates with the presence or absence of TDP-43 aggregates in a sample or a particular body part or body region referred to herein above may comprise: - determining the amount of the compound bound to the TDP-43 aggregates; - correlating the amount of compound bound to TDP-43 aggregates with the amount of TDP-43 aggregates in the sample or in a particular body part or body region; - optionally comparing the amount of compound bound to TDP-43 aggregates in the sample or in a particular body part or body region with a normal control value in a healthy control subject. Includes.

[0380] If the amount of compound bound to TDP-43 aggregates is higher than the normal control value for healthy / reference subjects, this indicates that the patient / subject is suffering from or at risk of developing a disease, disorder, or condition associated with TDP-43 aggregates. Specifically, if the amount of compound bound to TDP-43 aggregates is higher than that expected in an individual who does not show clinical evidence of a neurodegenerative disease, it can be assumed that the patient has a predisposition to a disease, disorder, or condition associated with TDP-43 aggregates or to having a TDP-43 proteinopathy.

[0381] Monitoring disease progression: In one embodiment, the present invention relates to a method for monitoring the progression of a disease, disorder, or condition associated with TDP-43 aggregates or a TDP-43 proteinopathy in a patient. Typically, the patient is undergoing or has undergone treatment for a disease, disorder, or condition associated with TDP-43 aggregates or a TDP-43 proteinopathy. In particular, the treatment may involve administration of an anti-TDP-43 medicament.

[0382] 1. A method of collecting data for monitoring the progression of a disease, disorder, or condition associated with TDP-43 aggregates, or for monitoring the progression of a TDP-43 proteinopathy in a patient, comprising: (a) contacting a sample, specific body part, or body region suspected of containing TDP-43 aggregates with a compound of the present invention; (b) binding the compound of the present invention to TDP-43 aggregates; (c) detecting the compound of the present invention bound to TDP-43 aggregates; (d) optionally correlating the presence or absence of a compound of the invention bound to TDP-43 aggregates with the presence or absence of TDP-43 aggregates in the sample or in a particular body part or region; and (e) optionally repeating steps (a) through (c), and, if present, optional step (d), at least once. Includes.

[0383] Steps (a) through (c), and optional step (d), if present, may be repeated one or more times to monitor the progression of a disease, disorder, or condition associated with TDP-43 aggregates or a TDP-43 proteinopathy over time. Preferably, the steps should be repeated until no further progression of the disease is observed in the patient.

[0384] Optionally, the step of correlating the presence or absence of a compound bound to TDP-43 aggregates with the presence or absence of TDP-43 aggregates in a sample or a particular body part or body region referred to herein above may comprise: - determining the amount of the compound bound to the TDP-43 aggregates; - correlating the amount of compound bound to TDP-43 aggregates with the amount of TDP-43 aggregates in the sample or in a particular body part or body region; - optionally comparing the amount of compound bound to TDP-43 aggregates in the sample or in a particular body part or body region with a normal control value in a healthy control subject. Includes.

[0385] In methods for monitoring progression over time, the amount of a compound of the invention bound to TDP-43 aggregates can optionally be compared at various time points during treatment, e.g., before and after initiation of treatment, and / or at various time points after initiation of treatment. A change, particularly a decrease, in the amount of a compound of the invention bound to TDP-43 aggregates can indicate that the disease is not progressing.

[0386] Responsiveness prediction: In one embodiment, the present invention relates to a method for predicting the responsiveness of a patient suffering from a disease, disorder or disorder associated with TDP-43 aggregates or suffering from a TDP-43 proteinopathy to treatment of said disease, disorder or disorder associated with TDP-43 aggregates or TDP-43 proteinopathy.

[0387] The method can be used to predict the most suitable treatment for a patient. In particular, the treatment can involve the administration of an anti-TDP-43 medication.

[0388] A method for predicting the responsiveness of a patient suffering from a disease, disorder or abnormality associated with TDP-43 aggregates or suffering from a TDP-43 proteinopathy to treatment of said disease, disorder or abnormality associated with TDP-43 aggregates or TDP-43 proteinopathy comprises: (a) contacting a sample, a specific body part, or a body region suspected of containing TDP-43 aggregates with a compound of the invention disclosed herein, or a diagnostic composition comprising a compound of the invention; (b) binding the compound of the present invention to TDP-43 aggregates; (c) detecting the compound of the present invention bound to TDP-43 aggregates; (d) optionally correlating the presence or absence of a compound of the invention bound to TDP-43 aggregates with the presence or absence of TDP-43 aggregates in the sample or in a particular body part or region; and (e) optionally repeating steps (a) through (c), and, if present, optional step (d), at least once. may include:

[0389] Typically, the patient is undergoing / has undergone treatment for a disease, disorder, or disorder associated with TDP-43 aggregates or a TDP-43 proteinopathy. In particular, the treatment may involve the administration of a medicament suitable for treating a disease, disorder, or disorder associated with TDP-43 aggregates.

[0390] This method allows for prediction of a patient's responsiveness to a certain treatment. In one embodiment, responsiveness can be estimated, for example, by repeating steps (a) to (c) and, if present, optional step (d), and monitoring the amount of the compound of the present invention bound to TDP-43 aggregates over time while the patient is undergoing treatment for a disease, disorder, or abnormality associated with TDP-43 aggregates or TDP-43 proteinopathy. If the amount changes over time, one skilled in the art can infer whether the patient is responsive to the treatment. Typically, if the amount of the compound of the present invention bound to TDP-43 aggregates decreases over time, the patient can be assumed to be responsive to the treatment. Typically, if the amount of the compound bound to TDP-43 aggregates remains essentially constant or increases over time, the patient can be assumed to be non-responsive to the treatment.

[0391] Alternatively, responsiveness can be estimated by determining the amount of a compound of the present invention bound to TDP-43 aggregates. The amount of a compound bound to TDP-43 aggregates can be compared to a control value, such as a normal control value, a preclinical control value, or a clinical control value. The control value can refer to a control value of a healthy control subject. Alternatively, the control value can refer to a control value of a subject known to be responsive to a treatment, or the control value can refer to a control value of a subject known to be non-responsive to a treatment. The outcome for responsiveness can be either "responsive" to a treatment, "non-responsive" to a treatment, or "response undetermined" to a treatment. Response to a treatment can vary from patient to patient.

[0392] Optionally, the step of correlating the presence or absence of a compound bound to TDP-43 aggregates with the presence or absence of TDP-43 aggregates in a sample or a particular body part or body region referred to herein above may comprise: - determining the amount of the compound bound to the TDP-43 aggregates; - correlating the amount of compound bound to TDP-43 aggregates with the amount of TDP-43 aggregates in the sample or in a particular body part or body region; - optionally comparing the amount of compound bound to TDP-43 aggregates in the sample or in a particular body part or body region with a normal control value in a healthy control subject. Includes.

[0393] The control value can be, for example, a normal control value, a preclinical control value, and / or a clinical control value. A "healthy control subject" or "healthy subject" is an individual who does not exhibit clinical evidence of a neurodegenerative disease.

[0394] If the amount of compound bound to TDP-43 aggregates in any of the methods summarized above is higher than the normal control value, the patient can be predicted to be suffering from or susceptible to a disease, disorder or condition associated with TDP-43 aggregates or a TDP-43 proteinopathy.

[0395] Any of the compounds of the present invention may be used in the methods summarized above. Preferably, the detectably labeled compounds of the present invention disclosed herein are used in the methods summarized above.

[0396] In a fourth aspect, the present invention relates to the use of the compounds of the present invention as diagnostic agents or tools for TDP-43 aggregates. In one embodiment, the present invention relates to the use of the compounds of the present invention as in vitro analytical standards or in vitro screening tools. The compounds of the present invention are also useful in in vivo diagnostic methods. In such instances, the compounds of the present invention may be detectably labeled or may contain non-radioactive isotopes.

[0397] In another embodiment, the present invention further relates to the use of the compounds of the present invention, more particularly the detectably labeled compounds of the present invention as defined herein, as diagnostic biomarkers, allowing for more efficient and accurate patient selection, for example for longitudinal monitoring in clinical studies or to aid in the development of new therapeutics for treating TDP-43 proteinopathy. In another embodiment, the present invention further relates to the use of the compounds of the present invention, more particularly the detectably labeled compounds of the present invention as defined herein, as biomarkers for TDP-43 aggregates or TDP-43 proteinopathy biomarkers.

[0398] In another embodiment, the compounds of the present invention can be used for research applications, particularly as analytical tools or reference molecules. The compounds can also be used for in vitro or in vivo detection of TDP-43 aggregates. The compounds of the present invention can be used to stain TDP-43 aggregates. For example, the compounds of the present invention can be used for histochemical detection in postmortem tissues, such as brain tissues. The compounds of the present invention are preferably detectably labeled compounds, and can be directly or indirectly labeled as discussed herein.

[0399] Kit of Parts In a fifth aspect, the present invention further relates to a kit for use in one or more of the methods of the present invention, the kit comprising a compound of the present invention as described herein. The kit typically comprises a container holding a compound of the present invention and instructions for use of the compound of the present invention. Preferably, the kit comprises a compound of formula (I) as disclosed herein. More preferably, the compound of the present invention is a detectably labeled compound (e.g., a compound of formula (IT) or (IF)).

[0400] The term "kit" generally refers to any diagnostic kit known in the art. More specifically, the latter term refers to the diagnostic kit described in Zrein et al., Clin. Diagn. Lab. Immunol., 1998, 5, 45-49.

[0401] The dosage of the detectably labeled compounds of the present invention will vary depending on the exact compound administered, the patient's weight, the size and type of sample, and other variables apparent to a practitioner skilled in the art. Generally, dosages preferably range from 0.001 μg / kg to 10 μg / kg, preferably from 0.01 μg / kg to 1.0 μg / kg. The radiation dose can be, for example, from 100 to 600 MBq, more preferably from 150 to 450 MBq.

[0402] In particular, such kits can be useful for performing the methods of the present invention (e.g., including, but not limited to, imaging, diagnostic, and monitoring methods), e.g., for diagnosing a disease, disorder, or condition associated with TDP-43 aggregates, or a TDP-43 proteinopathy. Such kits can include all the necessary components for performing the methods provided herein. Typically, each component is stored separately in a single overall package. Suitable additional components for inclusion in the kit are, for example, buffers, detectable dyes, laboratory equipment, reaction vessels, instructions, etc. The instructions for use can be tailored to the particular method in which the kit is to be used.

[0403] The present invention further relates to a kit for preparing a detectably labeled compound of the present invention, in particular, where the detectable label is a radioisotope. Thus, the kit includes a detectably labeled precursor of the compound of formula (I) and a labeling agent that reacts with the precursor to introduce a detectable (e.g., radioactive) label. Preferred precursors are compounds of formulas (II), (III), and (IV). The labeling agent that reacts with the precursor introduces a detectable (e.g., radioactive) label, e.g., 18 F or 3 The labeling agent can be an agent that induces H. 18F-can be a fluorinating agent.

[0404] Methods for preparing compounds of the present invention In a sixth aspect, the present invention further relates to a process for preparing a compound of formula (I).

[0405] Non-radioactive isotopic compounds: In one embodiment, the present invention provides a process for preparing a compound of formula (I), as described above, comprising: The compound of formula (II) is reacted with R 10 to obtain a compound of formula (I)

[0406] [ka]

[0407] (In the formula, n, R 1 and R 2 is as defined above, R 10 is a 5- or 6-membered carbocyclic or heterocyclic ring optionally substituted with F, NH, CN and / or CH, wherein the heterocyclic ring contains one or more heteroatoms selected from N, O and S, and wherein the 5- or 6-membered carbocyclic or heterocyclic ring is substituted with Br or I. The present invention relates to a method comprising:

[0408] In a preferred embodiment, R 10 is the following group:

[0409] [ka]

[0410] (wherein X is N and R 5 is CH3 or H, and Hal is Br or I) or

[0411] [ka]

[0412] wherein Hal is Br; R 10 teeth,

[0413] [ka]

[0414] wherein Hal is Br or I; R 3 is F and R 4 is NH2 and R 7 is H and R 8 is H, R 3 is NH2 and R 4 is F and R 7 is H and R 8 is H, R 3 is CN and R 4 is NH2 and R 7 is H and R 8 is H, R 3 is H and R 4 is NH2 and R 7 is H and R 8 is CN, R 3 is H and R 4 is NH2 and R 7 is H and R 8 is F, or R 3 is H and R 4 is NH2 and R 7 is H and R 8 is CN),

[0415] [ka]

[0416] (In the formula, R 9is selected from H, F, NH2 or CH3, and Hal is Br or I,

[0417] [ka]

[0418] wherein Hal is Br,

[0419] [ka]

[0420] wherein Hal is Br,

[0421] [ka]

[0422] wherein Hal is Br,

[0423] [ka]

[0424] wherein Hal is Br,

[0425] [ka]

[0426] wherein Hal is Br, and

[0427] [ka]

[0428] wherein Hal is Br. is.

[0429] The compound having formula (II) is reacted with R10 The reaction can be carried out by any suitable method. In one alternative, the reaction can be carried out in the presence of a diamine chelating agent such as DMEDA, a base such as potassium carbonate, a catalyst such as CuI, and an aprotic solvent such as dioxane. In another alternative, the reaction can be carried out under Pd-coupling conditions in the presence of a Pd catalyst such as Pd[P(Ph)] or Pd(OAc), XantPhos.

[0430] Tritium ( 3 H) a compound detectably labeled with In one embodiment, the present invention provides a method for the detection of tritium ( 3 3. A method for preparing a compound of the present invention having formula (IT) detectably labeled with tritium (H), comprising the step of radiolabeling a precursor of formula (IT) with a radioactive isotope, wherein at least one leaving group of the precursor of the compound having formula (IT) is tritium ( 3 H).

[0431] At least one to three hydrogens (H) are each replaced by tritium ( 3 H), which has been replaced by tritium ( 3 Preferably, compounds having formula (IT) detectably labeled with tritium (H) are defined as compounds having formula (IT) detectably labeled with tritium (H). 3 H), which has been replaced by tritium ( 3 More preferably defined are compounds having formula (IT) detectably labeled with three hydrogens (H). 3 H), which has been replaced by tritium ( 3 Even more preferably defined are compounds having formula (I) detectably labeled with H).

[0432] In another embodiment, the present invention provides a method for preparing a compound of formula (IT), comprising: By exchange of either Br or I with T, by use of T and a suitable catalyst, or by introduction of a CT group, a compound of formula (III)

[0433] [ka]

[0434] (In the formula, Z is selected from C—Br, CI and CH; n is 1 or 2, R 1 is H, hydroxy(C1-C4)alkyl or F, preferably R 1 is H or F, R 11 is a 5- or 6-membered carbocyclic or heterocyclic ring optionally substituted with Br, I, F, NH, CN and / or CH, the heterocyclic ring containing one or more heteroatoms selected from N, O and S; Z and / or R 11 At least one of the above contains Br or I, at least one of Br or I is replaced by or is T, or R 11 contains an NH moiety, the NH-moiety is replaced with N-CT3, T is 3 H) or a stereoisomer, polymorph, racemic mixture, tautomer, pharmaceutically acceptable salt, prodrug, hydrate or solvate thereof, or a mixture thereof, is reacted with a precursor compound having T (i.e. 3 H) radiolabeling step The present invention provides a method comprising:

[0435] In a preferred embodiment, Z is C—Br or CI, where Br or I is replaced by T.

[0436] In another preferred embodiment, R 11 contains Br or I, and Br or I is replaced by T.

[0437] In a preferred embodiment, Z is C—Br or CI, where Br or I is replaced by T, and R 11 contains Br or I, and Br or I is replaced by T.

[0438] In another embodiment, Z is CH and R 11 teeth

[0439] [ka]

[0440] (In the formula, R 5 is H or CH3, and R 11 where at least one H is replaced by CT3).

[0441] In another embodiment, the present invention provides a method for preparing a compound of formula (IT), comprising: Formula (III)

[0442] [ka]

[0443] (In the formula, n is 1 or 2, preferably 2 R 1 HA-COOR A and R A is (C1-C4) alkyl, preferably hydroxyethyl or hydroxymethyl, more preferably hydroxymethyl, and R 1 At least one hydrogen atom in is replaced by T, preferably R 1 Two of the hydrogen atoms in are replaced by T, R 6 is T or H, T is 3 H, R 11is a 5- or 6-membered carbocyclic or heterocyclic ring optionally substituted with F, NH, CN and / or CH, the heterocyclic ring containing one or more heteroatoms selected from N, O and S, preferably N. The precursor compound having T (i.e. 3 H).

[0444] Preferably, tritium ( 3 H), detectably labeled with formula (IT), wherein T is 3 The compound having H is

[0445] [ka]

[0446] Includes.

[0447] Preferably, the precursor having formula (III) according to the present invention is

[0448] [ka]

[0449] may be selected from:

[0450] Radioactive isotopes, e.g. 3 Methods used to introduce H are well known in the art and include those described below.

[0451] [ka]

[0452] In this scheme, the substituents Br, NH, F and CN are shown by way of example only, and the definitions of formula (1-T) apply in this regard.

[0453] Further examples are shown in the scheme below.

[0454] [ka]

[0455] Further examples are shown in the scheme below.

[0456] [ka]

[0457] [ka]

[0458] means that the nitrogen atom may be present at any available position in the ring.

[0459] Further examples are shown in the scheme below.

[0460] [ka]

[0461] Regarding the introduction of T (tritium), 3 The H radiolabeling agent can be tritium gas. This method can be carried out in the presence of a catalyst, such as palladium on carbon (Pd / C) or Lindlar catalyst, a solvent, such as N,N-dimethylformamide (DMF), and a base, such as N,N-diisopropylethylamine (DIEA). Alternatively, 3 The 3 H radiolabeling agent can be LiT, prepared from n-BuLi and tritium gas in the presence of AlCl3 and a solvent such as THF in the presence of TMEDA.

[0462] In one embodiment, the present invention provides a method for preparing a precursor compound of formula (III), as described above, comprising the steps of: The compound of formula (II) as defined above is 10followed by either NBS bromination or acid cleavage of the trimethylsilylethoxymethyl (SEM)-protecting group to give a compound of formula (Ia). The following examples are given by way of illustration:

[0463] [ka]

[0464] (In the formula, n and R 1 is as defined above, R 10 is a 5- or 6-membered carbocyclic or heterocyclic ring optionally substituted with F, NH, CN and / or CH, wherein the heterocyclic ring contains one or more heteroatoms selected from N, O and S, and wherein the 5- or 6-membered carbocyclic or heterocyclic ring is substituted with Br or I and optionally substituted with SEM).

[0465] In one preferred embodiment, R 10 is the following group:

[0466] [ka]

[0467] (wherein X is N and R 5 is SEM and Hal is Br or I),

[0468] [ka]

[0469] wherein Hal is Br; R 10 teeth,

[0470] [ka]

[0471] wherein Hal is Br or I; R 3 is F and R 4 is NH2 and R 7 is H and R 8 is H, R 3 is NH2 and R 4 is F and R 7 is H and R 8 is H, R 3 is CN and R 4 is NH2 and R 7 is H and R 8 is H, R 3 is H and R 4 is NH2 and R 7 is H and R 8 is a CN, or R 3 is H and R 4 is NH2 and R 7 is H and R 8 is F, R 3 is H and R 4 is NH2 and R 7 is H and R 8 is CN),

[0472] [ka]

[0473] (In the formula, R 9 is selected from H, F, NH or CH, and when Cl is present, Hal is Br, or when Br is present, Hal is I;

[0474] [ka]

[0475] wherein Hal is Br when Cl is present, or Hal is I when Br is present, and

[0476] [ka]

[0477] wherein Hal is Br when Cl is present, or I when Br is present;

[0478] [ka]

[0479] wherein Hal is Br when Cl is present, or I when Br is present;

[0480] [ka]

[0481] wherein Hal is Br when Cl is present, or I when Br is present;

[0482] [ka]

[0483] wherein Hal is Br when Cl is present, or Hal is I when Br is present, and

[0484] [ka]

[0485] wherein Hal is Br when Cl is present, or I when Br is present. is.

[0486] The compound having formula (II) is reacted with R 10 The reaction can be carried out by any suitable method. In one alternative, the reaction can be carried out in the presence of a diamine chelating agent such as DMEDA, a base such as potassium carbonate, a catalyst such as CuI, and an aprotic solvent such as dioxane. In another alternative, the reaction can be carried out under Pd-coupling conditions in the presence of a Pd catalyst such as Pd[P(Ph)] or Pd(OAc), XantPhos.

[0487] Fluorine ( 18 F) A compound detectably labeled with: In one embodiment, the present invention provides a method for treating fluorine ( 18 F) A method for preparing a detectably labeled compound of the present invention, comprising the steps of:

[0488] [ka]

[0489] (In the formula, n and R 2 is as defined above, R 14 In the radiolabeling step, 18 is a leaving group (LG) that is displaced by F) The precursor having the radioisotope [ 18 F].

[0490] In a preferred embodiment, the present invention provides a fluorine ( 18 F), a method for preparing a compound of the invention detectably labeled with formula (IV):

[0491] [ka]

[0492] (In the formula, n is 1 or 2, preferably 1; R2 teeth

[0493] [ka]

[0494] (In the formula, R 9 is H), R 14 In the radiolabeling step, 18 is a leaving group displaced by F) The precursor having the radioisotope [ 18 F].

[0495] In another embodiment, the present invention provides a method for treating fluorine ( 18 F), a method for preparing a compound of the present invention detectably labeled with formula (V):

[0496] [ka]

[0497] (In the formula, n, R 1 and R 2 is as defined herein for compounds of formula (I), R 14 In the radiolabeling step, 18 A precursor having a leaving group (LG) that is replaced by F is treated with a radioisotope [ 18 F].

[0498] In a preferred embodiment, the present invention provides a fluorine ( 18 F), a method for preparing a compound of the present invention detectably labeled with formula (V):

[0499] [ka]

[0500] (wherein n is 1 or 2, preferably 2; R 1 is H or hydroxy(C1-C4)alkyl, preferably hydroxyethyl or hydroxymethyl, more preferably hydroxymethyl; R 2 is a 5- or 6-membered carbocyclic or heterocyclic ring optionally substituted with F, NH, CN and / or CH, the heterocyclic ring containing one or more heteroatoms selected from N, O and S, preferably N, and preferably R 2 teeth,

[0501] [ka]

[0502] (In the formula, R 9 is H), R 14 In the radiolabeling step, 18 A precursor having a leaving group (LG) that is replaced by F is treated with a radioisotope [ 18 F].

[0503] Fluorination is carried out by K[ 18 F], Cs 18 F, Na 18 F, Rb 18 F, Kryptofix

[0222] K 18 F, 18 F Tetra (C 1~6 alkyl)ammonium salts and [ 18 F] tetrabutylammonium fluoride 18 It may be carried out in the presence of an F-fluorinating agent.

[0504] Preferably, the leaving group (LG) is C 1~4 Alkyl sulfonate or C 6~10More preferably, the leaving group (LG) is mesylate, tosylate, or nosylate or nitro. Even more preferably, the leaving group (LG) is mesylate or nitro.

[0505] 18 Suitable solvents for the F-fluorination step are known to those skilled in the art. The solvent may be selected from the group consisting of, for example, DMF, DMSO, acetonitrile, DMA, or a mixture thereof. Preferably, the solvent is acetonitrile or DMSO.

[0506] Preferably, fluorine ( 18 F) A method for preparing detectably labeled compound 1 is to prepare a precursor L1 in which the leaving group (LG), in this case mesylate, is substituted as shown below: 18 F-fluorinating agents, such as K[ 18 F] or [ 18 In the presence of [F]TBAF, fluorine ( 18 F) is replaced by a radiolabeling step.

[0507] [ka]

[0508] Preferably, fluorine ( 18 F) A method for preparing detectably labeled compound 17 is to prepare a precursor 20 in which the leaving group (LG), in this case nitro, is substituted as shown below: 18 F-fluorinating agents, such as K[ 18 F] or [ 18 In the presence of [F]TBAF, fluorine ( 18 F) is replaced by a radiolabeling step.

[0509] [ka]

[0510] The compounds of the present invention can be prepared by one of the general methods illustrated in the following schemes, which are presented for illustrative purposes only and should not be construed as limiting.

[0511] Precursor compounds having formula (II), (III), (IV) or (V) as defined above, or stereoisomers, polymorphs, racemic mixtures, tautomers, pharmaceutically acceptable salts, prodrugs, hydrates or solvates thereof, are part of the present invention.

[0512] [Table 1A]

[0513] [Table 1B]

[0514] General synthetic scheme: Synthetic scheme for preparing 2-[2-fluoro-6-[(3R)-3-fluoropyrrolidin-1-yl]-3-pyridyl]-5-(3-pyridyl)-6,7-dihydrothiazolo[5,4-c]pyridin-4-one (compound 1)

[0515] [ka]

[0516] Synthetic scheme for preparing 2-(2-fluoro-6-(4-(hydroxymethyl)piperidin-1-yl)pyridin-3-yl)-5-(pyridin-3-yl)-6,7-dihydrothiazolo[5,4-c]pyridin-4(5H)-one (compound 17)

[0517] [ka]

[0518] [ 3 [H] Synthetic scheme for preparing precursor compounds 5-(3-amino-2,6-dibromo-4-fluorophenyl)-2-(5-bromo-6-(pyrrolidin-1-yl)pyridin-3-yl)-6,7-dihydrothiazolo[5,4-c]pyridin-4(5H)-one ( 3 Synthetic scheme for preparing H (precursor of compound 1)

[0519] [ka]

[0520] Ethyl 1-(3-bromo-5-(5-(5-bromopyridin-3-yl)-4-oxo-4,5,6,7-tetrahydrothiazolo[5,4-c]pyridin-2-yl)-6-fluoropyridin-2-yl)piperidine-4-carboxylate ( 3 Synthetic scheme for preparing H (precursor of compound 17)

[0521] [ka]

[0522] 3 H-labeled compounds can be prepared from suitable precursor compounds containing halogen atoms by catalytic tritium dehalogenation using tritium gas (M. Saljoughian, Synthesis (2002), pp. 1781-1801) or by the addition of methyl iodide [ 3 from a suitable precursor compound containing an NH moiety by methylation with [H] or 3 They can be prepared from suitable precursor compounds having a moiety that is susceptible to reduction by a [H]-containing reducing agent (Y. Chen, Chemistry 25(2019):3405-3439). Preferably, 3 The solvent used for 3H-labeling is DMF or DMA, preferably the solvent is DMF.

[0523] 18 Synthetic scheme for preparing precursors for F-labeling [(3S)-1-[6-fluoro-5-[4-oxo-5-(3-pyridyl)-6,7-dihydrothiazolo[5,4-c]pyridin-2-yl]-2-pyridyl]pyrrolidin-3-yl]methanesulfonate( 18 Synthetic scheme for preparing F (precursor of compound 1)

[0524] [ka]

[0525] 2-(6-(4-(hydroxymethyl)piperidin-1-yl)-2-nitropyridin-3-yl)-5-(pyridin-3-yl)-6,7-dihydrothiazolo[5,4-c]pyridin-4(5H)-one ( 18 Synthetic scheme for preparing F (precursor of compound 17)

[0526] [ka]

[0527] The reaction is carried out in the presence of a fluorinating agent and typically a solvent.

[0528] 18 The F-labeled compound is a precursor compound containing LG, 18 F-fluorinating agent, resulting in LG 18 It can be prepared by replacing by F. 18 F-fluorinating agent is 18 Tetraalkylammonium salts of F (e.g. 18 F Tetra (C 1~6 alkyl) ammonium salts, e.g. 18 F]tetrabutylammonium fluoride), 18 Tetraalkylphosphonium salts of F (e.g. 18 F Tetra (C 1~6 Alkyl)phosphonium salt), K[ 18 F], Cs 18 F, Na 18 F, Rb 18F or Kryptofix

[0222] K 18 F. Preferably, 18 F-fluorinating agent is Cs 18 F, K 18 F or [ 18 F] tetrabutylammonium fluoride. 18 Reagents, solvents and conditions that can be used for F-fluorination are well known to those skilled in the art (L. Cai, S. Lu, V. Pike, Eur. J. Org. Chem. 2008, pp. 2853-2873; J. Fluorine Chem., 27(1985): pp. 177-191; Schubiger PA, Friebe M., Lehmann L. (eds.), PET-Chemistry - The Driving Force in Molecular Imaging. Springer, Berlin Heidelberg, Coenen, Fluorine-18 Labeling Methods: Features and Possibilities of Basic Reactions, (2006) pp. 15-50). Preferably, 18 The solvent used for F-fluorination is DMF, DMSO, acetonitrile, DMA, or a mixture thereof, preferably the solvent is acetonitrile or DMSO.

[0529] as a radioactive label 18 Although the reaction for F is shown above, other radiolabels can be introduced following a similar procedure.

[0530] This invention is illustrated by the following examples which should not be construed as limiting. [Example]

[0531] All reagents and solvents were obtained from commercial sources and used without further purification. 1H NMR spectra were recorded on a JEOL-500 MHz NMR spectrometer. Mass spectra (MS) were recorded on a UPLC H-Class Plus with a photodiode array detector and a QDa mass spectrometer. Chromatography was performed using silica gel (Fluka: Silica gel 60, 0.063-0.2 mm) and appropriate solvents as indicated in the specific examples. Flash purification was performed on a Biotage Isolera One flash purification system using HP-Sil or KP-NH SNAP cartridges (Biotage) and solvent gradients as indicated in the specific examples. Thin-layer chromatography (TLC) was performed on silica gel plates with UV detection.

[0532] Although some of the examples do not indicate that each compound is detectably labeled, the corresponding detectably labeled compound is contemplated, e.g., a detectably labeled starting material, e.g., C( 3 H)3, ( 11 C) H3 or 18 It is understood that it can be readily prepared by using F-containing starting materials.

[0533] Example 1 Synthesis of 2-[2-fluoro-6-[(3R)-3-fluoropyrrolidin-1-yl]-3-pyridyl]-5-(3-pyridyl)-6,7-dihydrothiazolo[5,4-c]pyridin-4-one

[0534] [ka]

[0535] Step-1: Synthesis of tert-butyl 3-bromo-2,4-dioxopiperidine-1-carboxylate (B) tert-Butyl 2,4-dioxopiperidine-1-carboxylate A (10 g, 46.9 mmol) was dissolved in carbon tetrachloride (125 mL) and cooled to 0 to 5 °C. To the above solution, N-bromosuccinimide (8.35 g, 46.9 mmol) was added portionwise, and stirring was continued at 28 °C for 1 h. The reaction mixture was diluted twice with ethyl acetate (500 mL) and water (200 mL). The organic phase was separated, dried over NaSO, filtered, and the solvent was removed under reduced pressure to give the title compound B as a white solid (10 g, 73%). 1 H NMR (500 MHz, DMSO-d6): δ 8.1 (s, 2H), 3.89 (t, 2H), 2.76(t, 2H), 1.45 (s, 9H). LCMS (ESI) 292.04 m / z [M+H] + .

[0536] Step-2: Synthesis of tert-butyl 2-amino-4-oxo-6,7-dihydrothiazolo[5,4-c]pyridine-5(4H)-carboxylate (C) The title compound B (10 g, 34.36 mmol), thiourea (2.61 g, 34.36 mmol), and sodium bicarbonate (2.88 g, 34.36 mmol) were dissolved in ethanol (160 mL) and heated in an oil bath at 80° C. for 2.5 hours. The reaction mixture was diluted twice with ethyl acetate (500 mL) and water (200 mL). The organic phase was separated, dried over NaSO, filtered, and the solvent was removed under reduced pressure. The resulting solid was recrystallized from ethanol to give the title compound C as a white solid (6.6 g, 71%). 1 H NMR (500 MHz, DMSO-d6): δ 8.1 (s, 2H), 3.89 (t, 2H), 2.76(t, 2H), 1.45 (s, 9H). LCMS (ESI) 270.3 m / z [M+H] + .

[0537] Step-3: Synthesis of tert-butyl 2-bromo-4-oxo-6,7-dihydrothiazolo[5,4-c]pyridine-5(4H)-carboxylate (D) The title compound C (6.6 g, 24.53 mmol) was dissolved in acetonitrile (82 mL) and cooled to −10° C. with stirring in an ice bath. tert-Butyl nitrite (4.3 mL, 36.4 mmol) was added to the above solution, and stirring was continued at −10° C. for 1 hour. Copper(II) bromide (6.5 g, 29.43 mmol) was added to the above mixture, which was stirred at 28° C. for 1 hour. The reaction mixture was basified to pH 8-9 with saturated aqueous sodium bicarbonate and filtered. The collected filtrate was diluted three times with ethyl acetate (300 mL) and water (100 mL). The organic phase was separated, dried over NaSO, filtered, and the solvent was removed under reduced pressure. The resulting residue was purified by column chromatography on silica gel (100-200 mesh) using an ethyl acetate / n-hexane gradient (0 / 100→10 / 90) to afford the title compound D as a white solid (4.73 g, 58%). 1 H NMR (500 MHz DMSO-d6): δ 4.12 (t, 2H), 3.10 (t, 2H), 1.5 (s, 9H). LCMS (ESI) 279 m / z [M+H-C4H8] + .

[0538] Step-4: Synthesis of tert-butyl 2-(2,6-difluoropyridin-3-yl)-4-oxo-6,7-dihydrothiazolo[5,4-c]pyridine-5(4H)-carboxylate (F) tert-Butyl 2-bromo-4-oxo-6,7-dihydrothiazolo[5,4-c]pyridine-5(4H)-carboxylate D (0.3 g, 0.9 mmol) was dissolved in 1,4-dioxane (15 mL) and degassed by passing a stream of nitrogen through the mixture. Then, [1,1'-bis(diphenylphosphino)ferrocene]-dichloropalladium(II) complex with dichloromethane (0.07 g, 0.09 mmol), the title compound E (0.21 g, 1.35 mmol), and tribasic potassium phosphate (0.28 g, 1.35 mmol) were added, and the reaction mixture was heated in an oil bath at 100 °C for 8 h. The reaction mixture was concentrated under reduced pressure. The residue was purified by column chromatography on silica gel (basic, 100-200 mesh) using an ethyl acetate / n-hexane gradient (0 / 100 → 10 / 90 → 15 / 85) to give the title compound F as a pale yellow solid (0.2 g, 60%). 1 H NMR (500 MHz DMSO-d6): δ 8.91 (q, 1H), 7.43 (dd, 1H), 4.1 (t, 2H), 3.19 (t, 2H), 1.5 (s, 9H). LCMS (ESI) 311.95 m / z [M+H-C4H8] + .

[0539] Step-5: Synthesis of tert-butyl (R)-2-(2-fluoro-6-(3-fluoropyrrolidin-1-yl)pyridin-3-yl)-4-oxo-6,7-dihydrothiazolo[5,4-c]pyridine-5(4H)-carboxylate (G) The title compound F (0.3 g, 0.86 mmol), (R)-3-fluoropyrrolidine hydrochloride (0.213 g, 1.7 mmol), and triethylamine (0.33 mL, 2.36 mmol) were suspended in n-butanol (8 mL) using a microwave vial. The sealed vial was then heated at 160° C. for 1 hour using a CEM microwave. The reaction mixture was concentrated under reduced pressure. The resulting residue was suspended in water (20 mL) and filtered on Whatman filter paper. The solid was washed with water (20 mL), followed by n-hexane (20 mL), and dried under reduced pressure to give the title compound G as an off-white solid (0.27 g, 98%). 1 H NMR (500 MHz, DMSO-d6): δ 8.75 (d, 1H), 8.06 (dd, 1H), 7.81 (s, 1H), 6.64 (d, 1H), 5.48 (d, 1H), 3.9-3.7 (m, 3H), 3.51-3.48 (m, 3H), 2.98 (t, 2H), 2.34.2.13 (m, 2H). LCMS (ESI) 319.00 m / z [M+H] + .

[0540] Step-6: Synthesis of (R)-2-(2-fluoro-6-(3-fluoropyrrolidin-1-yl)pyridin-3-yl)-6,7-dihydrothiazolo[5,4-c]pyridin-4(5H)-one (H) The title compound G (0.188 g, 0.011 mmol) was dissolved in DCM (4 mL) and cooled to 0 °C with stirring in an ice bath. 4 M HCl in 1,4-dioxane (2 mL) was added to the above solution, and stirring was continued at RT for 3 h. After completion of the reaction, the solvent was removed under reduced pressure. The resulting residue was dissolved in ice-cold water and basified with saturated aqueous sodium bicarbonate to pH 8-9. The precipitated solid was filtered and dried to give the title compound H as a pale yellow solid (0.135 g, 92%). 1H NMR (500 MHz DMSO-d6): δ 7.81 (d, 2H), 7.75 (s, 1H), 6.66 (d, 2H), 5.48 (d, 1H), 3.52 (m, 6H), 2.96 (t, 2H), 2.24 (m, 2H). LCMS (ESI) 318 [M+H]+.

[0541] Step-7: Synthesis of (R)-2-(2-fluoro-6-(3-fluoropyrrolidin-1-yl)pyridin-3-yl)-5-(pyridin-3-yl)-6,7-dihydrothiazolo[5,4-c]pyridin-4(5H)-one (1) Compound H (0.05 g, 0.14 mmol) and 3-iodopyridine 8 (0.09 g, 0.44 mmol) were dissolved in degassed 1,4-dioxane (7.5 mL). After the addition of Pd[PPh3]4 (0.008 g, 0.0074 mmol), XantPhos (0.01 g, 0.014 mmol), and Cs2CO3 (0.14 g, 0.44 mmol), the reaction mixture was heated in a sand bath at approximately 100 °C for 24 h. The reaction mixture was concentrated under reduced pressure to give a crude residue. The residue was purified by column chromatography on silica gel (100-200 mesh) using a CHCl2 / MeOH gradient (100 / 0 → 99.5 / 0.5 → 99 / 1) to give 1 as a yellow solid (0.025 g, 41%). 1 H NMR (400 MHz DMSO-d6): δ 8.67 (d, 1H), 8.43 (m, 2H), 7.87 (m, 1H), 7.5 (dd, 1H), 6.64 (dd, 1H), 5.49 (d, 1H), 4.17 (t, 2H), 3.72 (m, 3H), 3.52 (m, 1H), 3.26 (t, 2H), 2.22 (m, 2H). LCMS (ESI) 414.1 m / z [M+H] +

[0542] Example 2 Synthesis of 5-(3-amino-2,6-dibromo-4-fluorophenyl)-2-(5-bromo-6-(pyrrolidin-1-yl)pyridin-3-yl)-6,7-dihydrothiazolo[5,4-c]pyridin-4(5H)-one

[0543] [ka]

[0544] Step-1: Synthesis of tert-butyl 3-bromo-2,4-dioxopiperidine-1-carboxylate (B) tert-Butyl 2,4-dioxopiperidine-1-carboxylate A (10 g, 46.9 mmol) was dissolved in carbon tetrachloride (125 mL) and cooled to 0 to 5 °C. To the above solution, N-bromosuccinimide (8.35 g, 46.9 mmol) was added portionwise, and stirring was continued at 28 °C for 1 h. The reaction mixture was diluted twice with ethyl acetate (500 mL) and water (200 mL). The organic phase was separated, dried over NaSO, filtered, and the solvent was removed under reduced pressure to give the title compound B as a white solid (10 g, 73%). 1 H NMR (500 MHz, DMSO-d6): δ 8.1 (s, 2H), 3.89 (t, 2H), 2.76(t, 2H), 1.45 (s, 9H). LCMS (ESI) 292.04 m / z [M+H] + .

[0545] Step-2: Synthesis of tert-butyl 2-amino-4-oxo-6,7-dihydrothiazolo[5,4-c]pyridine-5(4H)-carboxylate (C) The title compound B (10 g, 34.36 mmol), thiourea (2.61 g, 34.36 mmol), and sodium bicarbonate (2.88 g, 34.36 mmol) were dissolved in ethanol (160 mL) and heated in an oil bath at 80° C. for 2.5 hours. The reaction mixture was diluted twice with ethyl acetate (500 mL) and water (200 mL). The organic phase was separated, dried over NaSO, filtered, and the solvent was removed under reduced pressure. The resulting solid was recrystallized from ethanol to give the title compound C as a white solid (6.6 g, 71%). 1 H NMR (500 MHz, DMSO-d6): δ 8.1 (s, 2H), 3.89 (t, 2H), 2.76(t, 2H), 1.45 (s, 9H). LCMS (ESI) 270.3 m / z [M+H] + .

[0546] Step-3: Synthesis of tert-butyl 2-bromo-4-oxo-6,7-dihydrothiazolo[5,4-c]pyridine-5(4H)-carboxylate (D) The title compound C (6.6 g, 24.53 mmol) was dissolved in acetonitrile (82 mL) and cooled to −10° C. with stirring in an ice bath. tert-Butyl nitrite (4.3 mL, 36.4 mmol) was added to the above solution, and stirring was continued at −10° C. for 1 hour. Copper(II) bromide (6.5 g, 29.43 mmol) was added to the above mixture, which was stirred at 28° C. for 1 hour. The reaction mixture was basified to pH 8-9 with saturated aqueous sodium bicarbonate and filtered. The collected filtrate was diluted three times with ethyl acetate (300 mL) and water (100 mL). The organic phase was separated, dried over NaSO, filtered, and the solvent was removed under reduced pressure. The resulting residue was purified by column chromatography on silica gel (100-200 mesh) using an ethyl acetate / n-hexane gradient (0 / 100→10 / 90) to afford the title compound D as a white solid (4.73 g, 58%). 1 H NMR (500 MHz DMSO-d6): δ 4.12 (t, 2H), 3.10 (t, 2H), 1.5 (s, 9H). LCMS (ESI) 279 m / z [M+H-C4H8]+ .

[0547] Step-4: Synthesis of tert-butyl 2-(2,6-difluoropyridin-3-yl)-4-oxo-6,7-dihydrothiazolo[5,4-c]pyridine-5(4H)-carboxylate (3) (F) tert-Butyl 2-bromo-4-oxo-6,7-dihydrothiazolo[5,4-c]pyridine-5(4H)-carboxylate D (0.3 g, 0.9 mmol) was dissolved in 1,4-dioxane (15 mL) and degassed by passing a stream of nitrogen through the mixture. Then, [1,1'-bis(diphenylphosphino)ferrocene]-dichloropalladium(II) complex with dichloromethane (0.07 g, 0.09 mmol), the title compound E (0.21 g, 1.35 mmol), and tribasic potassium phosphate (0.28 g, 1.35 mmol) were added, and the reaction mixture was heated in an oil bath at 100 °C for 8 h. The reaction mixture was concentrated under reduced pressure. The residue was purified by column chromatography on silica gel (basic, 100-200 mesh) using an ethyl acetate / n-hexane gradient (0 / 100 → 10 / 90 → 15 / 85) to give the title compound F as a pale yellow solid (0.2 g, 60%). 1 H NMR (500 MHz DMSO-d6): δ 8.91 (q, 1H), 7.43 (dd, 1H), 4.1 (t, 2H), 3.19 (t, 2H), 1.5 (s, 9H). LCMS (ESI) 311.95 m / z [M+H-C4H8] + .

[0548] Step-5: Synthesis of tert-butyl (R)-2-(2-fluoro-6-(3-fluoropyrrolidin-1-yl)pyridin-3-yl)-4-oxo-6,7-dihydrothiazolo[5,4-c]pyridine-5(4H)-carboxylate (G) The title compound F (0.3 g, 0.86 mmol), (R)-3-fluoropyrrolidine hydrochloride (0.213 g, 1.7 mmol), and triethylamine (0.33 mL, 2.36 mmol) were suspended in n-butanol (8 mL) using a microwave vial. The sealed vial was then heated at 160° C. for 1 hour using a CEM microwave. The reaction mixture was concentrated under reduced pressure. The resulting residue was suspended in water (20 mL) and filtered on Whatman filter paper. The solid was washed with water (20 mL), followed by n-hexane (20 mL), and dried under reduced pressure to give the title compound G as an off-white solid (0.27 g, 98%). 1 H NMR (500 MHz, DMSO-d6): δ 8.75 (d, 1H), 8.06 (dd, 1H), 7.81 (s, 1H), 6.64 (d, 1H), 5.48 (d, 1H), 3.9-3.7 (m, 3H), 3.51-3.48 (m, 3H), 2.98 (t, 2H), 2.34.2.13 (m, 2H). LCMS (ESI) 319.00 m / z [M+H] + .

[0549] Step-6: Synthesis of (R)-2-(2-fluoro-6-(3-fluoropyrrolidin-1-yl)pyridin-3-yl)-6,7-dihydrothiazolo[5,4-c]pyridin-4(5H)-one (H) The title compound G (0.188 g, 0.011 mmol) was dissolved in DCM (4 mL) and cooled to 0 °C with stirring in an ice bath. 4 M HCl in 1,4-dioxane (2 mL) was added to the above solution, and stirring was continued at RT for 3 h. After completion of the reaction, the solvent was removed under reduced pressure. The resulting residue was dissolved in ice-cold water and basified with saturated aqueous sodium bicarbonate to pH 8-9. The precipitated solid was filtered and dried to give the title compound H as a pale yellow solid (0.135 g, 92%). 1H NMR (500 MHz DMSO-d6): δ 7.81 (d, 2H), 7.75 (s, 1H), 6.66 (d, 2H), 5.48 (d, 1H), 3.52 (m, 6H), 2.96 (t, 2H), 2.24 (m, 2H). LCMS (ESI) 318 [M+H] + .

[0550] Step-7: Synthesis of (R)-5-(5-bromopyridin-3-yl)-2-(2-fluoro-6-(3-fluoropyrrolidin-1-yl)pyridin-3-yl)-6,7-dihydrothiazolo[5,4-c]pyridin-4(5H)-one (I) (R)-2-(2-Fluoro-6-(3-fluoropyrrolidin-1-yl)pyridin-3-yl)-6,7-dihydrothiazolo[5,4-c]pyridin-4(5H)-one H (0.05 g, 0.14 mmol) and 3,5-dibromopyridine 7 (0.1 g, 0.44 mmol) were dissolved in degassed 1,4-dioxane (5 mL). After the addition of Pd[PPh] (0.008 g, 0.0074 mmol), XantPhos (0.08 g, 0.014 mmol), and CsCO (0.1 g, 0.3 mmol), the reaction mixture was heated in an oil bath at approximately 100 °C for 24 h. The reaction mixture was concentrated under reduced pressure to give a crude residue. The residue was purified by column chromatography on silica gel (100-200 mesh) using a CH2Cl2 / MeOH gradient (100 / 0 → 99 / 1 → 98.8 / 1.2) to give I as a yellow solid (0.05 g, 68%). 1 H NMR (400 MHz DMSO-d6): δ 8.67 (d, 1H), 8.58 (d, 1H), 8.41 (dd, 1H), 6.64 (dd, 1H), 5.48 (d, 1H), 4.18 (t, 2H), 3.74 (m, 3H), 3.52(m, 1H), 3.25 (t, 2H), 2.22 (m, 2H). LCMS (ESI) 492.0 m / z [M+H] +

[0551] Step-8: Synthesis of (R)-2-(5-bromo-2-fluoro-6-(3-fluoropyrrolidin-1-yl)pyridin-3-yl)-5-(5-bromopyridin-3-yl)-6,7-dihydrothiazolo[5,4-c]pyridin-4(5H)-one (2) The title compound I (0.14 g, 0.28 mmol) was dissolved in DMF (5 mL) and cooled to -50 °C. NBS (0.1 g, 0.56 mmol) was added portionwise. The reaction mixture was allowed to warm to 20 °C over a period of 1 h. The reaction mixture was diluted with cold water (50 mL) and the solid was filtered. The crude solid was purified by preparative TLC plate using dichloromethane / methanol (98.5 / 1.5) as the mobile phase to give the title compound 2 as a pale yellow solid (0.083 g, 52%). 1 H NMR (400 MHz DMSO-d6): δ 8.67 (d, 1H), 8.59 (d, 1H), 8.57 (d, 1H), 8.18 (t, 1H), 5.44 (d, 1H), 4.19 (t, 2H), 3.99 (m, 4H), 3.26(t, 2H), 2.17 (m, 2H). LCMS (ESI) 571.55 m / z [M+2H] + .

[0552] Example 3 Synthesis of (S)-1-(6-fluoro-5-(4-oxo-5-(pyridin-3-yl)-4,5,6,7-tetrahydrothiazolo[5,4-c]pyridin-2-yl)pyridin-2-yl)pyrrolidin-3-yl methanesulfonate

[0553] [ka]

[0554] Step-1: Synthesis of tert-butyl 3-bromo-2,4-dioxopiperidine-1-carboxylate (B) tert-Butyl 2,4-dioxopiperidine-1-carboxylate A (10 g, 46.9 mmol) was dissolved in carbon tetrachloride (125 mL) and cooled to 0 to 5 °C. To the above solution, N-bromosuccinimide (8.35 g, 46.9 mmol) was added portionwise, and stirring was continued at 28 °C for 1 h. The reaction mixture was diluted twice with ethyl acetate (500 mL) and water (200 mL). The organic phase was separated, dried over NaSO, filtered, and the solvent was removed under reduced pressure to give the title compound B as a white solid (10 g, 73%). 1 H NMR (500 MHz, DMSO-d6): δ 8.1 (s, 2H), 3.89 (t, 2H), 2.76(t, 2H), 1.45 (s, 9H). LCMS (ESI) 292.04 m / z [M+H] + .

[0555] Step-2: Synthesis of tert-butyl 2-amino-4-oxo-6,7-dihydrothiazolo[5,4-c]pyridine-5(4H)-carboxylate (C) The title compound B (10 g, 34.36 mmol), thiourea (2.61 g, 34.36 mmol), and sodium bicarbonate (2.88 g, 34.36 mmol) were dissolved in ethanol (160 mL) and heated in an oil bath at 80° C. for 2.5 hours. The reaction mixture was diluted twice with ethyl acetate (500 mL) and water (200 mL). The organic phase was separated, dried over NaSO, filtered, and the solvent was removed under reduced pressure. The resulting solid was recrystallized from ethanol to give the title compound C as a white solid (6.6 g, 71%). 1 H NMR (500 MHz, DMSO-d6): δ 8.1 (s, 2H), 3.89 (t, 2H), 2.76(t, 2H), 1.45 (s, 9H). LCMS (ESI) 270.3 m / z [M+H] + .

[0556] Step-3: Synthesis of tert-butyl 2-bromo-4-oxo-6,7-dihydrothiazolo[5,4-c]pyridine-5(4H)-carboxylate (D) The title compound C (6.6 g, 24.53 mmol) was dissolved in acetonitrile (82 mL) and cooled to −10° C. with stirring in an ice bath. tert-Butyl nitrite (4.3 mL, 36.4 mmol) was added to the above solution, and stirring was continued at −10° C. for 1 hour. Copper(II) bromide (6.5 g, 29.43 mmol) was added to the above mixture, which was stirred at 28° C. for 1 hour. The reaction mixture was basified to pH 8-9 with saturated aqueous sodium bicarbonate and filtered. The collected filtrate was diluted three times with ethyl acetate (300 mL) and water (100 mL). The organic phase was separated, dried over NaSO, filtered, and the solvent was removed under reduced pressure. The resulting residue was purified by column chromatography on silica gel (100-200 mesh) using an ethyl acetate / n-hexane gradient (0 / 100→10 / 90) to afford the title compound D as a white solid (4.73 g, 58%). 1 H NMR (500 MHz DMSO-d6): δ 4.12 (t, 2H), 3.10 (t, 2H), 1.5 (s, 9H). LCMS (ESI) 279 m / z [M+H-C4H8] + .

[0557] Step-4: Synthesis of tert-butyl 2-(2,6-difluoropyridin-3-yl)-4-oxo-6,7-dihydrothiazolo[5,4-c]pyridine-5(4H)-carboxylate (3) (F) tert-Butyl 2-bromo-4-oxo-6,7-dihydrothiazolo[5,4-c]pyridine-5(4H)-carboxylate D (0.3 g, 0.9 mmol) was dissolved in 1,4-dioxane (15 mL) and degassed by passing a stream of nitrogen through the mixture. Then, [1,1'-bis(diphenylphosphino)ferrocene]-dichloropalladium(II) complex with dichloromethane (0.07 g, 0.09 mmol), the title compound E (0.21 g, 1.35 mmol), and tribasic potassium phosphate (0.28 g, 1.35 mmol) were added, and the reaction mixture was heated in an oil bath at 100 °C for 8 h. The reaction mixture was concentrated under reduced pressure. The residue was purified by column chromatography on silica gel (basic, 100-200 mesh) using an ethyl acetate / n-hexane gradient (0 / 100 → 10 / 90 → 15 / 85) to give the title compound F as a pale yellow solid (0.2 g, 60%). 1 H NMR (500 MHz DMSO-d6): δ 8.91 (q, 1H), 7.43 (dd, 1H), 4.1 (t, 2H), 3.19 (t, 2H), 1.5 (s, 9H). LCMS (ESI) 311.95 m / z [M+H-C4H8] + .

[0558] Step-5: Synthesis of tert-butyl (S)-2-(2-fluoro-6-(3-hydroxypyrrolidin-1-yl)pyridin-3-yl)-4-oxo-6,7-dihydrothiazolo[5,4-c]pyridine-5(4H)-carboxylate (J) The title compound 3 (0.2 g, 0.54 mmol), (S)-pyrrolidin-3-ol hydrochloride F (0.068 g, 0.54 mmol), and N,N-diisopropylethylamine (0.09 mL, 1.06 mmol) were suspended in DMSO (20 mL) and stirred at room temperature for 3 h. The reaction mixture was quenched with cold water (200 mL), and the precipitated solid was filtered on Whatman filter paper and dried to give the title compound J as a pale yellow solid (0.22 g, 92%).

[0559] Step-6: Synthesis of (S)-2-(2-fluoro-6-(3-hydroxypyrrolidin-1-yl)pyridin-3-yl)-6,7-dihydrothiazolo[5,4-c]pyridin-4(5H)-one (K) The title compound J (0.22 g, 0.5 mmol) was dissolved in DCM (4.4 mL) and cooled to 0 °C with stirring in an ice bath. 4 M HCl in 1,4-dioxane (2.2 mL) was added to the above solution, and stirring was continued at RT for 2 h. After completion of the reaction, the solvent was removed under reduced pressure. The resulting residue was dissolved in ice-cold water and basified with saturated aqueous sodium bicarbonate to pH 8-9. The precipitated solid was filtered and dried to give the title compound K as a pale yellow solid (0.155 g, 92%). 1 H NMR (500 MHz DMSO-d6): δ 8.33 (t, 1H), 7.8 (s, 1H), 6.55 (d, 1H), 5.06 (s, 1H), 4.41 (s, 1H), 3.54 (m, 6H), 2.99 (t, 2H), 1.99 (d, 2H). LCMS (ESI) 334.9 [M+H] + .

[0560] Step-7: Synthesis of (S)-2-(6-(3-((tert-butyldimethylsilyl)oxy)pyrrolidin-1-yl)-2-fluoropyridin-3-yl)-6,7-dihydrothiazolo[5,4-c]pyridin-4(5H)-one (L) The title compound K (0.15 g, 0.44 mmol) and imidazole (0.3 g, 4.5 mmol) dissolved in DMF (3 mL) were cooled to 0 °C in an ice bath. tert-Butyldimethylsilyl chloride (0.5 g, 3.35 mmol) was added portionwise. The reaction mixture was stirred at room temperature for 36 h. The reaction mixture was diluted with cold water (150 mL), and the precipitated solid was filtered and dried to give the title compound L as a pale yellow solid (0.177 g, 88%). 1H NMR (400 MHz, DMSO-d6): δ 8.34 (t, 1H), 7.81 (s, 1H), 6.57 (dd, 1H), 4.59 (s, 1H), 3.63 (dd, 1H), 3.5 (m, 4H), 3.36 (m, 2H), 3.09 (s, 3H), 2.99 (t, 2H), 2.01 (d, 2H), 0.86 (s, 9H), 0.1 (d, 6H). LCMS (ESI) 449.25 m / z [M+H]+

[0561] Step-8: Synthesis of (S)-2-(6-(3-((tert-butyldimethylsilyl)oxy)pyrrolidin-1-yl)-2-fluoropyridin-3-yl)-5-(pyridin-3-yl)-6,7-dihydrothiazolo[5,4-c]pyridin-4(5H)-one (M) Compound L (0.17 g, 0.37 mmol) and 3-iodopyridine 8 (0.23 g, 1.1 mmol) were dissolved in degassed 1,4-dioxane (17 mL). After the addition of Pd[PPh3]4 (0.043 g, 0.03 mmol), XantPhos (0.043 g, 0.075 mmol), and Cs2CO3 (0.37 g, 1.13 mmol), the reaction mixture was heated in an oil bath at approximately 100 °C for 24 h. The reaction mixture was concentrated under reduced pressure to give a crude residue. The crude residue was purified by column chromatography on silica gel (100-200 mesh) using a CHCl2 / MeOH gradient (100 / 0 → 99 / 1 → 98 / 2) to give M as a yellow solid (0.17 g, 89%). 1 H NMR (500 MHz, DMSO-d6): δ 8.75 (d, 1H), 8.54 (q, 1H), 8.48 (t, 1H), 7.94(m, 1H), 7.52(m, 1H), 6.7 (dd, 1H), 4.7 (s, 1H), 4.25 (t, LCMS (ESI) 526 m / z [M+H]+ .

[0562] Step-9: Synthesis of (S)-2-(2-fluoro-6-(3-hydroxypyrrolidin-1-yl)pyridin-3-yl)-5-(pyridin-3-yl)-6,7-dihydrothiazolo[5,4-c]pyridin-4(5H)-one (N) The title compound M (0.14 g, 0.26 mmol) was dissolved in DCM (2.8 mL) and cooled to 0 °C with stirring in an ice bath. 4 M HCl in 1,4-dioxane (1.4 mL) was added to the above solution, and stirring was continued at RT for 3 h. After completion of the reaction, the solvent was removed under reduced pressure. The resulting residue was dissolved in ice-cold water and basified with saturated aqueous sodium bicarbonate to pH 8-9. The precipitated solid was filtered and dried to give the title compound N as a pale yellow solid (0.08 g, 72%). 1 H NMR (500 MHz, DMSO-d6): δ 8.66 (d, 1H), 8.44 (q, 1H), 8.38 (t, 1H), 7.84(m, 1H), 7.47(m, 1H), 6.7 (d, 1H), 6.58 (d, 1H), 5.08 (s, 1H), 4.42 (s, 1H), 4.16 (t, 2H), 3.55 (m, 4H), 3.25 (t, 2H), 2.0 (d, 2H). LCMS (ESI) 411.9 m / z [M+H] + .

[0563] Step-10: Synthesis of ((S)-1-(6-fluoro-5-(4-oxo-5-(pyridin-3-yl)-4,5,6,7-tetrahydrothiazolo[5,4-c]pyridin-2-yl)pyridin-2-yl)pyrrolidin-3-yl methanesulfonate (3) The title compound N (0.07 g, 0.17 mmol) was dissolved in pyridine (2.3 mL) and cooled to −50° C. with stirring in a dry ice bath. Methanesulfonyl chloride (0.46 g, 4.08 mmol) was added to the above solution and stirring was continued at 20° C. for 1 h. The reaction mixture was diluted with water (35 mL), and the precipitated solid was filtered, washed with water (2×15 mL), and dried to give the title compound 3 as a pale yellow solid (0.06 g, 72%). 1 H NMR (500 MHz, DMSO-d6): δ 8.66 (d, 1H), 8.43 (m, 2H), 7.84 (m, 1H), 7.47 (q, 1H), 6.65 (dd, 1H), 5.45 (s, 1H), 4.16 (t, 2H), 3.76 (m, 3H), 3.55 (dd, 1H), 3.26 (m, 5H), 2.34 (s, 2H). LCMS (ESI) 489.85 m / z [M+H] + .

[0564] Example 4 Tritiated 2-[2-fluoro-6-[(3R)-3-fluoropyrrolidin-1-yl]-3-pyridyl]-5-(3-pyridyl)-6,7-dihydrothiazolo[5,4-c]pyridin-4-one 3 Synthesis of H-compound 1

[0565] [ka]

[0566] T is 3 It means H.

[0567] 2.49 mg of brominated precursor 2, 50 μL DIPEA, and 33.09 mg of Lindlar catalyst were suspended in 0.3 ml DMF. The suspension was degassed three times using a high-vacuum manifold and stirred overnight at room temperature under an atmosphere of tritium gas (772 mbar, 9.1 Ci). The solvent was removed in vacuo, and the labile tritium was exchanged by adding 0.3 ml of methanol, stirring the solution, and again removing the solvent in vacuo. This process was repeated twice. The thoroughly dried solid was extracted with 5 ml of ethanol / DMF (4:1), and the suspension was filtered through a 0.2 μm nylon membrane to obtain a clear solution. The following HPLC conditions were found to be suitable for the purification of the compound: Waters Sunfire C18, 10 × 250 mm; Solvent A: water + 0.1% TFA; Solvent B: acetonitrile + 0.1% TFA. Purified product 3 H-Compound 1 was obtained (SA 48.5 Ci / mmol, 99% purity).

[0568] Example 5 Fluorinated 2-[2-fluoro-6-[(3R)-3-fluoropyrrolidin-1-yl]-3-pyridyl]-5-(3-pyridyl)-6,7-dihydrothiazolo[5,4-c]pyridin-4-one 18 Synthesis of F compound 1

[0569] [ka]

[0570] [ in shipping vials (O-18 enriched water obtained from a commercial cyclotron facility) 18The [F] fluoride was transferred onto a Chromafix PS-HCO cartridge where it was collected (activated with KOTf (0.2 M, 6 mL) followed by water (6 mL)). It was then eluted with a solution of TBAOTf (6 mg in 0.6 mL) into the reaction vessel of a TRACERlab® module. The solution was first heated to 70 °C for 3.5 min under vacuum and helium flow, and the thermal treatment was continued for an additional 2.5 min at 95 °C under the same conditions. The reactor was then cooled to 60 °C. A solution of (S)-1-(6-fluoro-5-(4-oxo-5-(pyridin-3-yl)-4,5,6,7-tetrahydrothiazolo[5,4-c]pyridin-2-yl)pyridin-2-yl)pyrrolidin-3-yl methanesulfonate 3 (0.5 mg in 1 mL anhydrous acetonitrile) was added to the reaction vial, and the vessel was heated to 100 °C for 10 min. The reaction vessel was cooled to 60°C and the mixture was diluted with HPLC buffer (4 mL). Purification was performed by HPLC using a semi-preparative Phenomenex Gemini C18 column (5 μm, 250 × 10 mm) and elution with a mixture of acetonitrile / ammonium acetate solution (20 mM, 42 / 58, v / v) at a flow rate of 4 mL / min. The radiolabeled product was eluted from the SPE cartridge with 1.0 mL of ethanol into a formulation flask pre-filled with 10 mL of formulation base (10 mg of ascorbic acid in saline). The resulting solution was passed through a sterile 0.2 μm membrane filter into a sterile filter-vented vial. The final product 18 F - Compound 1 was assayed and a sample removed for QC testing. Confirmation of product identity was 19 F-Determined by simultaneous injection of samples of standard compounds.

[0571] Example Compounds 6 to 9 and 12 to 16 Following the coupling procedure reported for the preparation of Example 1, utilizing the components of Example 1, Step 6, Example 10, Step 2, Example 15, Step 2, and the halogen derivatives indicated in Table 1, the following compounds were prepared.

[0572] [Table 2A]

[0573] [Table 2B]

[0574] [Table 2C]

[0575] Example compounds 11, 21, 22, and 33 Following the coupling procedure reported for the preparation of Example 15 (Step 3), utilizing components from Example 1, Step 6, Example compound 10, Step 2, and the halogen derivatives indicated in Table 2, the following compounds were prepared.

[0576] [Table 3A]

[0577] [Table 3B]

[0578] Synthesis of 5-(3-amino-4-fluorophenyl)-2-(2-fluoro-6-(pyrrolidin-1-yl)pyridin-3-yl)-6,7-dihydrothiazolo[5,4-c]pyridin-4(5H)-one (Example Compound 10)

[0579] [ka]

[0580] Step-1: Synthesis of tert-butyl 2-(2-fluoro-6-(pyrrolidin-1-yl)pyridin-3-yl)-4-oxo-6,7-dihydrothiazolo[5,4-c]pyridine-5(4H)-carboxylate In a 20 mL microwave vial, the title compound D (0.3 g, 0.9 mmol) and commercially available (2-fluoro-6-(pyrrolidin-1-yl)pyridin-3-yl)boronic acid (0.38 g, 1.8 mmol) were dissolved in degassed DMA (10 mL). After the addition of Pd(OAc) (0.01 g, 0.046 mmol), dppf (0.05 g, 0.092 mmol), CuCl (0.089 g, 0.9 mmol), and CsCO (0.588 g, 1.66 mmol), the reaction mixture was heated in a sand bath at approximately 100 °C for 5 h. The above reaction was repeated once more. The reaction mixture from each run was diluted with CHCl (300 mL) and water / brine (140 mL; 1 / 1). The organic phase was separated, washed with brine (80 mL), dried over NaSO, filtered, and concentrated under reduced pressure. The dark residue was purified by chromatography on silica (100 g, HP-Ultra) using a Biotage Isolera system with a CHCl / EtOAc gradient (100 / 0 → 95 / 5 → 90 / 10 → 90 / 10) to give the crude coupled product. The crude coupled product was further purified by chromatography on silica (100 g, HP-Ultra) using a Biotage Isolera system with a CHCl / EtOAc gradient (100 / 0 → 95 / 5 → 90 / 10 → 90 / 10) to give the coupled product as a pale yellow solid (0.473 g, 63%). 1 H NMR (400 MHz, DMSO-d6 / CDCl3): δ 8.33 (t, 1H), 6.46 (dd, 1H), 4.06 (t, 2H), 3.08 (t, 2H), 2.00 (br-s, 4H), 1.50 (s, 9H). LCMS (ESI) 419 / 383.15 / 319.11 m / z [M+H] + .

[0581] Step-2: Synthesis of 2-(2-fluoro-6-(pyrrolidin-1-yl)pyridin-3-yl)-6,7-dihydrothiazolo[5,4-c]pyridin-4(5H)-one The starting material, tert-butyl 2-(2-fluoro-6-(pyrrolidin-1-yl)pyridin-3-yl)-4-oxo-6,7-dihydrothiazolo[5,4-c]pyridine-5(4H)-carboxylate (0.473 g, 1.13 mmol) was dissolved in CHCl (25 mL), and the mixture was cooled to 0 °C. TFA (4.4 mL) was added at 0 °C, and the reaction mixture was stirred at room temperature for 1 h. The solvent was evaporated under reduced pressure, and the residue was dissolved in CHCl (300 mL) and saturated NaHCO (80 mL). The organic phase was separated, washed with brine (80 mL), dried over NaSO, filtered, and concentrated under reduced pressure to give the deprotected product as a pale yellow solid (0.349 g, 97%). 1 H NMR (400 MHz, DMSO-d6): δ 8.33 (dd, 1H), 7.80 (br-s, 1H), 6.54 (dd, 1H), 3.53-3.43 (m, 6H), 2.99 (t, 2H), 1.97 (br-s, 4H). LCMS (ESI) 319.15 m / z [M+H] + .

[0582] Step-3: Synthesis of tert-butyl (5-bromo-2-fluorophenyl) (tert-butoxycarbonyl)carbamate Commercially available 5-bromo-2-fluoroaniline (0.5 g, 2.6 mmol) was dissolved in THF (15 mL) and BocO (2.29 g, 10.5 mmol) was added. After the addition of DMAP (0.482 g, 3.95 mmol), the reaction mixture was stirred at room temperature for 3 days. The reaction mixture was diluted with EtOAc (150 mL) and water / brine (80 mL; 1 / 1). The organic phase was separated, dried over NaSO, filtered, and concentrated under reduced pressure. The dark residue was purified by chromatography on silica (50 g, HP-Ultra) using a Biotage Isolera system with an EtOAc / n-heptane gradient (5 / 95 → 10 / 90 → 20 / 80 → 20 / 80) to give the bis-Boc-protected product as a colorless oil, which became a white solid (0.452 g, 44%) upon storage at room temperature. 1H NMR (400 MHz, DMSO-d6): δ 7.74-7.70 (m, 1H), 7.63-7.59 (m, 1H), 7.33 (t, 1H), 1.39 (s, 18H).

[0583] Step-4: Synthesis of tert-butyl (tert-butoxycarbonyl)(2-fluoro-5-(2-(2-fluoro-6-(pyrrolidin-1-yl)pyridin-3-yl)-4-oxo-6,7-dihydrothiazolo[5,4-c]pyridin-5(4H)-yl)phenyl)carbamate and tert-butyl (2-fluoro-5-(2-(2-fluoro-6-(pyrrolidin-1-yl)pyridin-3-yl)-4-oxo-6,7-dihydrothiazolo[5,4-c]pyridin-5(4H)-yl)phenyl)carbamate In a 5 mL microwave vial, Pd(OAc) (0.002 g, 0.01 mmol) and XantPhos (0.016 g, 0.02 mmol) were dissolved in degassed 1,4-dioxane (6 mL). The reaction mixture was heated in a sand bath at approximately 120 °C for approximately 2 min to form the catalyst. After the addition of 2-(2-fluoro-6-(pyrrolidin-1-yl)pyridin-3-yl)-6,7-dihydrothiazolo[5,4-c]pyridin-4(5H)-one (0.035 g, 0.11 mmol), tert-butyl (5-bromo-2-fluorophenyl)(tert-butoxycarbonyl)carbamate (0.107 g, 0.275 mmol), and CsCO (0.163 g, 0.5 mmol), the reaction mixture was heated in a sand bath at approximately 120 °C for 18 h. The reaction mixture was diluted with CHCl (50 mL) and water / brine (20 mL; 1 / 1). The organic phase was separated, washed with brine (20 mL), dried over NaSO, filtered, and concentrated under reduced pressure. The dark residue was purified by chromatography on silica (12 g, puriFlash, Interchim) using a Biotage Isolera system with an EtOAc / n-heptane gradient (0 / 95 → 40 / 60 → 80 / 20 → 100 / 0) to give a mixture of bis-Boc-protected and mono-Boc-protected coupling products as a yellow solid (0.0081 g, 11%). LCMS (ESI) m / z [M+H] 628.44 / 528.32 (bis-Boc) and 528.30 (mono-Boc) + .

[0584] Step-5: Synthesis of 5-(3-amino-4-fluorophenyl)-2-(2-fluoro-6-(pyrrolidin-1-yl)pyridin-3-yl)-6,7-dihydrothiazolo[5,4-c]pyridin-4(5H)-one (Compound 10) The mixture of bis-Boc-protected and mono-Boc-protected coupling products (0.0225 g, 0.036 mmol) was dissolved in CHCl (5 mL), and the mixture was cooled to 0 °C. TFA (0.39 mL) was added at 0 °C, and the reaction mixture was stirred at room temperature for 3 h. The solvent was evaporated under reduced pressure, and the residue was dissolved in CHCl (100 mL) and saturated NaHCO (30 mL). The organic phase was separated, washed with brine (20 mL), dried over NaSO, filtered, and concentrated under reduced pressure. The residue was purified by chromatography on silica (12 g, puriFlash, Interchim) using a Biotage Isolera system with an EtOAc / n-heptane gradient (5 / 95 → 40 / 60 → 80 / 20 → 80 / 20) to elute impurities, followed by EtOAc / CH2Cl2 (50 / 50) to give compound 10 as a pale amber solid (0.0084 g, 54%). 1 H NMR (400 MHz, DMSO-d6): δ 8.36 (t, 1H), 7.01 (dd, 1H), 6.77 (dd, 1H), 6.56 (dd, 1H), 6.52-6.48 (m, 1H), 5.24 (br-s, 2H), 3.99 (t, 2H), 3.48 (br-s, 4H), 3.19 (t, 2H), 1.99 (br-s, 4H). LCMS (ESI) 428.21 m / z [M+H] + .

[0585] Example compounds 26, 28 to 32, 34 Following the coupling procedure reported for the preparation of Example 10 (Steps 4-5), utilizing components from Example 1, Step 6, Example 15, Step 2, Example 16, Step 2, Example 17, Step 2, and the halogen derivatives indicated in Table 3, the following compounds were prepared.

[0586] [Table 4A]

[0587] [Table 4B]

[0588] [Table 4C]

[0589] [Table 4D]

[0590] Example Compounds 35 to 37 Following the coupling procedure reported for the preparation of Example 1, utilizing the components of Example 15, Step 2, Example 16, Step 2, and the halogen derivatives indicated in Table 4, the following compounds were prepared.

[0591] [Table 5A]

[0592] [Table 5B]

[0593] Example 6 Synthesis of 2-(2-fluoro-6-(4-(hydroxymethyl)piperidin-1-yl)pyridin-3-yl)-5-(pyridin-3-yl)-6,7-dihydrothiazolo[5,4-c]pyridin-4(5H)-one (Example Compound 17)

[0594] [ka]

[0595] Step-1: Synthesis of tert-butyl 3-bromo-2,4-dioxopiperidine-1-carboxylate (B) tert-Butyl 2,4-dioxopiperidine-1-carboxylate A (10 g, 46.9 mmol) was dissolved in CCl (125 mL), and the mixture was cooled to 0 to 5 °C. NBS (8.35 g, 46.9 mmol) was then added portionwise, and stirring was continued at 28 °C for 1 h. The reaction mixture was diluted with ethyl acetate (2 × 500 mL) and water (200 mL). The organic phase was separated, dried over NaSO, filtered, and the solvent was removed under reduced pressure to give the title compound B as a white solid (10 g, 73%). 1 H NMR (500 MHz, DMSO-d6): δ 8.1 (s, 2H), 3.89 (t, 2H), 2.76(t, 2H), 1.45 (s, 9H). LCMS (ESI) 292.04 m / z [M+H] + .

[0596] Step-2: Synthesis of tert-butyl 2-amino-4-oxo-6,7-dihydrothiazolo[5,4-c]pyridine-5(4H)-carboxylate (C) The title compound B (10 g, 34.36 mmol), thiourea (2.61 g, 34.36 mmol), and sodium bicarbonate (2.88 g, 34.36 mmol) were dissolved in EtOH (160 mL) and heated in an oil bath at 80 °C for 2.5 h. The reaction mixture was diluted with ethyl acetate (2 × 500 mL) and water (200 mL). The organic phase was separated, dried over NaSO, filtered, and the solvent was removed under reduced pressure. The solid was recrystallized from EtOH to give the title compound C as a white solid (6.6 g, 71%). 1 H NMR (500 MHz, DMSO-d6): δ 8.1 (s, 2H), 3.89 (t, 2H), 2.76(t, 2H), 1.45 (s, 9H). LCMS (ESI) 270.3 m / z [M+H] + .

[0597] Step-3: Synthesis of tert-butyl 2-bromo-4-oxo-6,7-dihydrothiazolo[5,4-c]pyridine-5(4H)-carboxylate (D) The title compound C (6.6 g, 24.53 mmol) was dissolved in CHCN (82 mL) and cooled to −10° C. with stirring in an ice bath. tert-Butyl nitrite (4.3 mL, 36.4 mmol) was added, and stirring was continued at −10° C. for 1 h. Copper(II) bromide (6.5 g, 29.43 mmol) was added, the ice bath was removed, and the reaction mixture was stirred at 28° C. for 1 h. The reaction mixture was basified to pH 8-9 with saturated aqueous sodium bicarbonate and filtered. The collected filtrate was diluted with ethyl acetate (3×300 mL) and water (100 mL). The organic phase was separated, dried over NaSO, filtered, and the solvent was removed under reduced pressure. The resulting residue was purified by column chromatography on silica gel (100-200 mesh) using an ethyl acetate / n-hexane gradient (0 / 100→10 / 90) to afford the title compound D as a white solid (4.73 g, 58%). 1 H NMR (500 MHz DMSO-d6): δ 4.12 (t, 2H), 3.10 (t, 2H), 1.5 (s, 9H). LCMS (ESI) 279 m / z [M+H-C4H8] + .

[0598] Step-4: Synthesis of tert-butyl 2-(2,6-difluoropyridin-3-yl)-4-oxo-6,7-dihydrothiazolo[5,4-c]pyridine-5(4H)-carboxylate (F) The title compound D (0.3 g, 0.9 mmol) was dissolved in 1,4-dioxane (15 mL) and degassed by passing a stream of nitrogen through the mixture. Pd(dppf)Cl × CHCl (0.07 g, 0.09 mmol), the title compound E (0.21 g, 1.35 mmol), and tribasic potassium phosphate (0.28 g, 1.35 mmol) were then added, and the reaction mixture was heated in an oil bath at 100 °C for 8 h. The reaction mixture was concentrated under reduced pressure. The residue was purified by column chromatography on silica gel (basic, 100-200 mesh) using an ethyl acetate / n-hexane gradient (0 / 100 → 10 / 90 → 15 / 85) to give the title compound F as a pale yellow solid (0.2 g, 60%). 1H NMR (500 MHz DMSO-d6): δ 8.91 (q, 1H), 7.43 (dd, 1H), 4.1 (t, 2H), 3.19 (t, 2H), 1.5 (s, 9H). LCMS (ESI) 311.95 m / z [M+H-C4H8] +.

[0599] Step-5: Synthesis of tert-butyl 2-(2-fluoro-6-(4-(hydroxymethyl)piperidin-1-yl)pyridin-3-yl)-4-oxo-6,7-dihydrothiazolo[5,4-c]pyridine-5(4H)-carboxylate (G) The title compound F (0.15 g, 0.41 mmol), piperidin-4-ylmethanol (0.045 g, 0.41 mmol), and DIPEA (0.09 g, 0.82 mmol) were dissolved in DMSO (15 mL) and stirred at room temperature for 2 h. After completion of the reaction, the reaction mixture was quenched with ice-cold water, filtered, and the solid was dried under reduced pressure to give the title compound G as an off-white solid (0.18 g, 98%). 1 H NMR (500 MHz, DMSO-d6): δ 8.32 (t, 1H), 6.91 (dd, 1H), 4.51 (t, 1H), 4.37 (d, 2H), 4.05 (t, 2H), 3.28 (t, 3H), 3.09 (t, 2H), 2.98 (t, 2H), 2.54 (s, 3H), 1.73 (m, 3H), 1.49 (s, 9H), 1.12 (m, 2H). LCMS (ESI) 463.20 m / z [M+H] + .

[0600] Step-6: Synthesis of 2-(2-fluoro-6-(4-(hydroxymethyl)piperidin-1-yl)pyridin-3-yl)-6,7-dihydrothiazolo[5,4-c]pyridin-4(5H)-one (H) The title compound G (0.18 g, 0.38 mmol) was dissolved in DCM (4 mL) and cooled to 0° C. with stirring in an ice bath. 4 M HCl in 1,4-dioxane (2 mL) was then added and stirring was continued at RT for 3 h. After completion of the reaction, the solvent was removed under reduced pressure. The residue was dissolved in ice-cold water and basified to pH 8-9 with saturated aqueous sodium bicarbonate. The solid was collected by filtration and dried under reduced pressure to give the title compound H as a pale yellow solid (0.13 g, 92%). 1 H NMR (500 MHz, DMSO-d6): δ 8.32 (t, 1H), 7.82 (s, 1H), 6.91 (dd, 1H), 4.51 (t, 1H), 4.36 (d, 2H), 3.5 (m, 2H), 3.27 (t, 2H), 2.97 (m, 4H), 1.73 (m, 3H), 1.12 (m, 2H). LCMS (ESI) 362.90 m / z [M+H] + .

[0601] Step-7: Synthesis of 2-(6-(4-(((tert-butyldimethylsilyl)oxy)methyl)piperidin-1-yl)-2-fluoropyridin-3-yl)-6,7-dihydrothiazolo[5,4-c]pyridin-4(5H)-one (I) The title compound H (0.13 g, 0.35 mmol) and imidazole (0.24 g, 3.6 mmol) were dissolved in DMF (3 mL), and the reaction mixture was cooled to 0 °C in an ice bath. tert-Butyldimethylsilyl chloride (0.26 g, 1.8 mmol) was then added portionwise, and the reaction mixture was stirred at RT for 24 h. The reaction mixture was diluted with cold water (50 mL), and the precipitate was collected by filtration and dried under reduced pressure. The residue was purified by column chromatography on silica gel (100-200 mesh) using a CHCl / MeOH gradient (100 / 0 → 99.5 / 0.5 → 99 / 1) to afford the title compound I as a yellow solid (0.15 g, 88%). 1H NMR (500 MHz, DMSO-d6): δ 8.32 (t, 1H), 7.82 (s, 1H), 6.90 (dd, 1H), 4.37 (d, 2H), 3.48 (m, 4H), 2.97 (m, 4H), 1.75 (d, 3H), 1.16 (m, 3H), 0.86 (s, 9H), 0.029 (s, 6H). LCMS (ESI) 477.00 m / z [M+H] + .

[0602] Step-8: Synthesis of 2-(6-(4-(((tert-butyldimethylsilyl)oxy)methyl)piperidin-1-yl)-2-fluoropyridin-3-yl)-5-(pyridin-3-yl)-6,7-dihydrothiazolo[5,4-c]pyridin-4(5H)-one (J) The title compound I (0.1 g, 0.21 mmol) and 3-iodopyridine (0.12 g, 0.63 mmol) were dissolved in degassed 1,4-dioxane (10 mL). After the addition of Pd[PPh3]4 (0.024 g, 0.021 mmol), XantPhos (0.024 g, 0.04 mmol), and Cs2CO3 (0.2 g, 0.63 mmol), the reaction mixture was heated in a sand bath at approximately 100 °C for 24 h. The reaction mixture was concentrated under reduced pressure. The residue was purified by column chromatography on silica gel (100-200 mesh) using a CHCl2 / MeOH gradient (100 / 0 → 99.5 / 0.5 → 99 / 1) to give J as a yellow solid (0.085 g, 73%). 1 H NMR (500 MHz, DMSO-d6): δ 8.32 (t, 1H), 7.82 (s, 1H), 6.90 (dd, 1H), 4.37 (d, 2H), 3.48 (m, 4H), 2.97 (m, 4H), 1.75 (d, 3H), 1.16 (m, 3H), 0.86 (s, 9H), 0.029 (s, 6H). LCMS (ESI) 477.00 m / z [M+H] + .

[0603] Step-9: Synthesis of 2-(2-fluoro-6-(4-(hydroxymethyl)piperidin-1-yl)pyridin-3-yl)-5-(pyridin-3-yl)-6,7-dihydrothiazolo[5,4-c]pyridin-4(5H)-one (Compound 17) The title compound J (0.08 g, 0.14 mmol) was dissolved in DCM (2 mL), and the mixture was cooled to 0 °C in an ice bath with stirring. 4 M HCl in 1,4-dioxane (1 mL) was then added, and stirring was continued at RT for 3 h. After completion of the reaction, the solvent was removed under reduced pressure. The residue was dissolved in ice-cold water and basified to pH 8-9 with saturated aqueous sodium bicarbonate. The precipitate was collected by filtration and dried under reduced pressure to give the title compound 4 as a pale yellow solid (0.058 g, 92%). 1 H NMR (500 MHz, DMSO-d6): δ 8.66 (d, 1H), 8.45 (q, 1H), 8.36 (dd, 1H), 7.84 (dt, 1H), 7.47 (q, 1H), 6.93 (dd, 1H), 4.52 (t, 1H), 4.38 (d, 2H), 4.16 (t, 2H), 3.26 (m, 4H), 2.98 (m, 2H), 1.72 (m, 3H), 1.13 (m, 2H). LCMS (ESI) 439.90 m / z [M+H] + .

[0604] Example 7 Ethyl 1-(6-fluoro-5-(4-oxo-5-(pyridin-3-yl)-4,5,6,7-tetrahydro-thiazolo[5,4-c]pyridin-2-yl)pyridin-2-yl)piperidine-4-carboxylate (compound 17) 3 Synthesis of H-precursor

[0605] [ka]

[0606] Step-1: Synthesis of tert-butyl 3-bromo-2,4-dioxopiperidine-1-carboxylate (B) tert-Butyl 2,4-dioxopiperidine-1-carboxylate A (10 g, 46.9 mmol) was dissolved in carbon tetrachloride (125 mL) and cooled to 0 to 5 °C. To the above solution, N-bromosuccinimide (8.35 g, 46.9 mmol) was added portionwise, and stirring was continued at 28 °C for 1 h. The reaction mixture was diluted twice with ethyl acetate (500 mL) and water (200 mL). The organic phase was separated, dried over NaSO, filtered, and the solvent was removed under reduced pressure to give the title compound B as a white solid (10 g, 73%). 1 H NMR (500 MHz, DMSO-d6): δ 8.1 (s, 2H), 3.89 (t, 2H), 2.76(t, 2H), 1.45 (s, 9H). LCMS (ESI) 292.04 m / z [M+H] + .

[0607] Step-2: Synthesis of tert-butyl 2-amino-4-oxo-6,7-dihydrothiazolo[5,4-c]pyridine-5(4H)-carboxylate (C) The title compound B (10 g, 34.36 mmol), thiourea (2.61 g, 34.36 mmol), and sodium bicarbonate (2.88 g, 34.36 mmol) were dissolved in ethanol (160 mL) and heated in an oil bath at 80° C. for 2.5 hours. The reaction mixture was diluted twice with ethyl acetate (500 mL) and water (200 mL). The organic phase was separated, dried over NaSO, filtered, and the solvent was removed under reduced pressure. The resulting solid was recrystallized from ethanol to give the title compound C as a white solid (6.6 g, 71%). 1 H NMR (500 MHz, DMSO-d6): δ 8.1 (s, 2H), 3.89 (t, 2H), 2.76(t, 2H), 1.45 (s, 9H). LCMS (ESI) 270.3 m / z [M+H] + .

[0608] Step-3: Synthesis of tert-butyl 2-bromo-4-oxo-6,7-dihydrothiazolo[5,4-c]pyridine-5(4H)-carboxylate (D) The title compound C (6.6 g, 24.53 mmol) was dissolved in acetonitrile (82 mL) and cooled to −10° C. with stirring in an ice bath. tert-Butyl nitrite (4.3 mL, 36.4 mmol) was added to the above solution, and stirring was continued at −10° C. for 1 hour. Copper(II) bromide (6.5 g, 29.43 mmol) was added to the above mixture, which was stirred at 28° C. for 1 hour. The reaction mixture was basified to pH 8-9 with saturated aqueous sodium bicarbonate and filtered. The collected filtrate was diluted three times with ethyl acetate (300 mL) and water (100 mL). The organic phase was separated, dried over NaSO, filtered, and the solvent was removed under reduced pressure. The resulting residue was purified by column chromatography on silica gel (100-200 mesh) using an ethyl acetate / n-hexane gradient (0 / 100→10 / 90) to afford the title compound D as a white solid (4.73 g, 58%). 1 H NMR (500 MHz DMSO-d6): δ 4.12 (t, 2H), 3.10 (t, 2H), 1.5 (s, 9H). LCMS (ESI) 279 m / z [M+H-C4H8] + .

[0609] Step-4: Synthesis of tert-butyl 2-(2,6-difluoropyridin-3-yl)-4-oxo-6,7-dihydrothiazolo[5,4-c]pyridine-5(4H)-carboxylate (F) tert-Butyl 2-bromo-4-oxo-6,7-dihydrothiazolo[5,4-c]pyridine-5(4H)-carboxylate D (0.3 g, 0.9 mmol) was dissolved in 1,4-dioxane (15 mL), and the mixture was degassed by passing a stream of nitrogen through the mixture. Then, [1,1'-bis(diphenylphosphino)ferrocene]-dichloropalladium(II) complex with dichloromethane (0.07 g, 0.09 mmol), the title compound E (0.21 g, 1.35 mmol), and tribasic potassium phosphate (0.28 g, 1.35 mmol) were added, and the reaction mixture was heated in an oil bath at 100 °C for 8 h. The reaction mixture was concentrated under reduced pressure. The residue was purified by column chromatography on silica gel (basic, 100-200 mesh) using an ethyl acetate / n-hexane gradient (0 / 100 → 10 / 90 → 15 / 85) to give the title compound F as a pale yellow solid (0.2 g, 60%). 1 H NMR (500 MHz DMSO-d6): δ 8.91 (q, 1H), 7.43 (dd, 1H), 4.1 (t, 2H), 3.19 (t, 2H), 1.5 (s, 9H). LCMS (ESI) 311.95 m / z [M+H-C4H8]+

[0610] Step-5: Synthesis of tert-butyl 2-(6-(4-(ethoxycarbonyl)piperidin-1-yl)-2-fluoropyridin-3-yl)-4-oxo-6,7-dihydrothiazolo[5,4-c]pyridine-5(4H)-carboxylate (K) The title compound F (0.3 g, 0.82 mmol), ethyl piperidine-4-carboxylate (0.13 g, 0.82 mmol), and DIPEA (0.2 g, 1.63 mmol) were dissolved in DMSO (30 mL), and the reaction mixture was stirred at room temperature for 2 hours. After completion of the reaction, the reaction mixture was quenched with ice-cold water, and the precipitated solid was filtered and dried to give the title compound K as an off-white solid (0.37 g, 90%). 1H NMR (500 MHz, DMSO-d6): δ 8.35 (t, 1H), 6.93 (dd, 1H), 4.26 (d, 2H), 4.07 (m, 4H), 3.15 (m, 2H), 3.10 (t, 2H), 2.69 (m, 1H), 1.93 (dd, 2H), 1.58 (m, 2H), 1.49 (s, 9H), 1.19 (t, 3H). LCMS (ESI) 505.35 m / z [M+H] + .

[0611] Step-6: Synthesis of ethyl 1-(6-fluoro-5-(4-oxo-4,5,6,7-tetrahydrothiazolo[5,4-c]pyridin-2-yl)pyridin-2-yl)piperidine-4-carboxylate (L) The title compound K (0.37 g, 0.73 mmol) was dissolved in DCM (11 mL), and the mixture was cooled to 0 °C in an ice bath with stirring. 4 M HCl in 1,4-dioxane (4.0 mL) was added to the above solution, and stirring was continued at RT for 2 h. After completion of the reaction, the solvent was removed under reduced pressure. The resulting residue was dissolved in ice-cold water and basified to pH 8-9 with saturated aqueous sodium bicarbonate. The precipitated solid was filtered and dried to give the title compound L as a pale yellow solid (0.26 g, 89%). 1 H NMR (500 MHz, DMSO-d6): δ 8.34 (t, 1H), 7.83 (s, 1H), 6.92 (dd, 1H), 4.25 (d, 2H), 4.08 (q, 2H), 3.50 (m, 2H), 3.12 (m, 2H), 2.99 (t, 2H), 2.69 (m, 1H), 1.93 (m, 2H), 1.57 (m, 2H), 1.21 (m, 3H). LCMS (ESI) 405.5 m / z [M+H] + .

[0612] Step-7: Synthesis of ethyl 1-(6-fluoro-5-(4-oxo-5-(pyridin-3-yl)-4,5,6,7-tetrahydrothiazolo[5,4-c]pyridin-2-yl)pyridin-2-yl)piperidine-4-carboxylate (18) The title compound L (0.15 g, 0.37 mmol) and 3-bromopyridine (0.26 g, 1.11 mmol) were dissolved in degassed 1,4-dioxane (15 mL). After the addition of Pd[PPh3]4 (0.021 g, 0.018 mmol), XantPhos (0.021 g, 0.037 mmol), and Cs2CO3 (0.24 g, 0.74 mmol), the reaction mixture was heated in an oil bath at approximately 100 °C for 24 h. The reaction mixture was concentrated under reduced pressure to give a crude residue. The crude residue was purified by column chromatography on silica gel (100-200 mesh) using a CHCl2 / MeOH gradient (100 / 0 → 99.5 / 0.5 → 99 / 1) to give compound 18 as a yellow solid (0.15 g, 75%). 1 H NMR (500 MHz, DMSO-d6): δ 8.67 (d, 1H), 8.58 (d, 1H), 8.38 (dd, 1H), 8.17 (t, 1H), 6.95 (dd, 1H), 4.26 (d, 2H), 4.18 (t, 2H), 4.08 (q, 2H), 3.25 (t, 2H), 3.16 (m, 2H), 2.72 (m, 1H), 1.94 (dd, 1H), 1.55 (m, 2H), 1.19 (t, 3H). LCMS (ESI) 560.30 m / z [M+H] + .

[0613] Example 8 2-(6-(4-(hydroxymethyl)piperidin-1-yl)-2-nitropyridin-3-yl)-5-(pyridin-3-yl)-6,7-dihydrothiazolo[5,4-c]pyridin-4(5H)-one (compound 17 18 Synthesis of F-precursor

[0614] [ka]

[0615] Step-1: Synthesis of tert-butyl 3-bromo-2,4-dioxopiperidine-1-carboxylate (B) tert-Butyl 2,4-dioxopiperidine-1-carboxylate A (10 g, 46.9 mmol) was dissolved in carbon tetrachloride (125 mL) and the mixture was cooled to 0 to 5 °C. To the above solution, N-bromosuccinimide (8.35 g, 46.9 mmol) was added portionwise and stirring was continued at 28 °C for 1 h. The reaction mixture was diluted twice with ethyl acetate (500 mL) and water (200 mL). The organic phase was separated, dried over NaSO, filtered, and the solvent was removed under reduced pressure to give the title compound B as a white solid (10 g, 73%). 1 H NMR (500 MHz, DMSO-d6): δ 8.1 (s, 2H), 3.89 (t, 2H), 2.76(t, 2H), 1.45 (s, 9H). LCMS (ESI) 292.04 m / z [M+H] + .

[0616] Step-2: Synthesis of tert-butyl 2-amino-4-oxo-6,7-dihydrothiazolo[5,4-c]pyridine-5(4H)-carboxylate (C) The title compound B (10 g, 34.36 mmol), thiourea (2.61 g, 34.36 mmol), and sodium bicarbonate (2.88 g, 34.36 mmol) were dissolved in ethanol (160 mL) and heated in an oil bath at 80° C. for 2.5 hours. The reaction mixture was diluted twice with ethyl acetate (500 mL) and water (200 mL). The organic phase was separated, dried over NaSO, filtered, and the solvent was removed under reduced pressure. The resulting solid was recrystallized from ethanol to give the title compound C as a white solid (6.6 g, 71%). 1 H NMR (500 MHz, DMSO-d6): δ 8.1 (s, 2H), 3.89 (t, 2H), 2.76(t, 2H), 1.45 (s, 9H). LCMS (ESI) 270.3 m / z [M+H] + .

[0617] Step-3: Synthesis of tert-butyl 2-(dibenzylamino)-4-oxo-6,7-dihydrothiazolo[5,4-c]pyridine-5(4H)-carboxylate (M) The title compound C (0.1 g, 0.37 mmol) and cesium bicarbonate (0.84 g, 2.6 mmol) were suspended in DMF (5 mL), and the mixture was cooled to 0 °C in an ice bath with stirring. Benzyl bromide (0.26 g, 1.5 mmol) was added to the above solution, and stirring was continued at RT for 7 h. The reaction mixture was quenched with water (50 mL) and extracted twice with ethyl acetate (25 mL). The organic phase was separated, dried over Na2SO4, filtered, and the solvent was removed under reduced pressure to give a residue. The residue was purified by column chromatography on basic silica gel (100-200 mesh) using a hexane / EtOAc gradient (100 / 0 → 98 / 2 → 96 / 4 → 94 / 6 → 92 / 8) to give the title compound M as an off-white sticky solid (0.09 g, 71%). 1 H NMR (400 MHz, DMSO-d6): δ 7.33 (m, 10H), 4.8 (s, 4H), 3.93 (t, 2H), 2.85 (t, 2H), 1.44 (s, 9H). LCMS (ESI) 448 m / z [MH] + .

[0618] Step-4: Synthesis of 2-(dibenzylamino)-6,7-dihydrothiazolo[5,4-c]pyridin-4(5H)-one (N) The title compound M (0.09 g, 0.2 mmol) was dissolved in DCM (2 mL) and cooled to 0 °C with stirring in an ice bath. TFA (0.45 mL) was added to the above solution, and stirring was continued at RT for 2 h. After completion of the reaction, the solvent was removed under reduced pressure. The resulting residue was dissolved in ice-cold water and basified to pH 8-9 with saturated aqueous sodium bicarbonate. The mixture was extracted twice with ethyl acetate (25 mL). The organic phase was separated, dried over Na2SO4, filtered, and the solvent was removed under reduced pressure to give the title compound N as an off-white solid (0.065 g, 93%). 1 H NMR (400 MHz, DMSO-d6): δ 7.32 (m, 10H), 4.76 (s, 4H), 3.37 (m, 2H), 2.73 (t, 2H). LCMS (ESI) 350.4 m / z [MH]+ .

[0619] Step-5: Synthesis of 2-(dibenzylamino)-5-(pyridin-3-yl)-6,7-dihydrothiazolo[5,4-c]pyridin-4(5H)-one (O) The title compound N (1.0 g, 2.86 mmol) and 3-iodopyridine (1.75 g, 8.5 mmol) were dissolved in degassed 1,4-dioxane (50 mL). After the addition of Pd[PPh3]4 (0.16 g, 0.14 mmol), XantPhos (0.16 g, 0.28 mmol), and Cs2CO3 (2.78 g, 8.5 mmol), the reaction mixture was heated in an oil bath at approximately 100 °C for 24 h. The reaction mixture was concentrated under reduced pressure to give a crude residue. The crude residue was purified by column chromatography on silica gel (100-200 mesh) using a CHCl2 / MeOH gradient (100 / 0 → 99.5 / 0.5 → 99 / 1 → 98 / 2) to give O as an off-white solid (0.75 g, 61%). 1 H NMR (400 MHz, DMSO-d6): δ 8.57 (d, 1H), 8.37 (dd, 1H), 7.74 (m, 1H), 7.39 (m, 5H), 7.31 (m, 6H), 4.81(s, 4H), 4.02 (t, 2H), 3.0 (t, 2H). LCMS (ESI) 424.8 m / z [MH] + .

[0620] Step-6: Synthesis of 2-amino-5-(pyridin-3-yl)-6,7-dihydrothiazolo[5,4-c]pyridin-4(5H)-one (P) The title compound O (0.75 g, 1.7 mmol) was suspended in toluene (75 mL) and cooled to 0 °C in an ice bath with stirring. Methanesulfonic acid (7.5 mL) was added to the above solution, and stirring was continued at 100 °C for 6 h. After completion of the reaction, the two layers were separated. The toluene phase was decanted to give a crude brown liquid. The crude liquid was cooled to 0 °C in an ice bath, quenched with saturated aqueous sodium bicarbonate to pH 8-9, and extracted with two volumes of 10% MeOH in DCM (75 mL). The organic phase was separated, dried over Na2SO4, filtered, and the solvent was removed under reduced pressure to give the title compound P as an off-white solid (0.22 g, 51%). 1 H NMR (400 MHz, DMSO-d6): δ 8.58 (d, 1H), 8.37 (d, 1H), 7.93 (s, 2H), 7.74 (m, 1H), 7.41 (d, 1H), 3.99 (t, 2H), 2.9 (t, 2H). LCMS (ESI) 244.8 m / z [MH] + .

[0621] Step-7: Synthesis of 2-bromo-5-(pyridin-3-yl)-6,7-dihydrothiazolo[5,4-c]pyridin-4(5H)-one (Q) The title compound P (0.5 g, 2.03 mmol) was dissolved in acetonitrile (25 mL) and cooled to 0° C. in an ice bath with stirring. tert-Butyl nitrite (0.7 mL, 6.0 mmol) was added to the above solution, and stirring was continued at 0° C. for 30 min. Copper(II) bromide (0.9 g, 4.06 mmol) was added to the above mixture, which was stirred at 28° C. for 3 h. The reaction mixture was basified to pH 8-9 with saturated aqueous sodium bicarbonate and filtered. The collected filtrate was diluted twice with 10% MeOH in DCM (100 mL). The organic phase was separated, dried over Na2SO4, filtered, and the solvent was removed under reduced pressure. The resulting residue was purified by column chromatography on silica gel (100-200 mesh) using a CH2Cl2 / MeOH gradient (100 / 0 → 99.5 / 0.5 → 99 / 1 → 98 / 2) to afford the title compound Q as an off-white solid (0.22 g, 35%). 1H NMR (400 MHz, DMSO-d6): δ 8.64 (m, 1H), 8.46 (dd, 1H), 7.83 (m, 1H), 7.48 (m, 1H), 4.13 ((t, 2H), 3.24 (t, 2H). LCMS (ESI) 307.7 m / z [MH] + .

[0622] Step-8: Synthesis of (1-(6-nitropyridin-2-yl)piperidin-4-yl)methanol (S) Compound R (2.5 g, 31.64 mmol), piperidin-4-ylmethanol (9.4 g, 81.7 mmol), and DIPEA (12.26 mL, 94.8 mmol) were suspended in dioxane (120 mL) using a microwave vial. The sealed vial was then heated at 100° C. for 3 hours using a CEM microwave. The reaction mixture was concentrated under reduced pressure to give a crude residue. The crude residue was purified by column chromatography on basic silica gel (100-200 mesh) using a hexane / EtOAc gradient (100 / 0 → 90 / 10 → 80 / 20 → 70 / 30 → 60 / 40 → 50 / 50) to give the title compound S as a yellow oil (4.0 g, 53%). 1 H NMR (500 MHz, DMSO-d6): δ 7.83 (t, 1H), 7.38 (d, 1H), 7.27 (d, 1H), 4.5 (t, 1H), 4.37 (d, 2H), 3.28 (t, 2H), 2.9 (m, 2H), 1.75 (d, 2H), 1.66 (m, 1H), 1.12 (m, 2H). LCMS (ESI) 238.1 m / z [M+H] + .

[0623] Step-9: Synthesis of (1-(5-bromo-6-nitropyridin-2-yl)piperidin-4-yl)methanol (T) The title compound S (2.1 g, 8.8 mmol) was dissolved in ACN (42 mL) and cooled to -30 °C. NBS (2.39 g, 13.29 mmol) was added portionwise. The reaction mixture was allowed to warm to 0 °C over a period of 2 h. The reaction mixture was quenched with water (50 mL) and extracted twice with ethyl acetate (100 mL). The organic phase was separated, dried over NaSO, filtered, and the solvent was removed under reduced pressure to give a crude residue. The crude residue was purified by column chromatography on basic silica gel (100-200 mesh) using a hexane / EtOAc gradient (100 / 0 → 90 / 10 → 80 / 20 → 70 / 30 → 60 / 40 → 50 / 50) to give the title compound T as a pale yellow oil (1.5 g, 54%). 1 H NMR (400 MHz, DMSO-d6): δ 7.96 (d, 1H), 7.11 (d, 1H), 4.5 (t, 1H), 4.23 (d, 2H), 3.26 (t, 2H), 2.88 (m, 2H), 1.68 (m, 3H),1.11 (m, 2H). LCMS (ESI) 313.9 m / z [MH] + .

[0624] Step-10: Synthesis of (1-(6-nitro-5-(trimethylstannyl)pyridin-2-yl)piperidin-4-yl)methanol (U) The title compound T (1.4 g, 4.4 mmol) was dissolved in degassed 1,4-dioxane (28 mL) in a microwave vial. After the addition of Pd[PPh3)2Cl2 (0.3 g, 0.44 mmol), hexamethyldistannane (1.3 mL, 1.95 g, 6.0 mmol), and triphenylarsine (0.135 g, 0.4 mmol), the vial was sealed and heated at 100 °C for 1 h in a CEM microwave. The reaction mixture was concentrated under reduced pressure to give a crude residue. The crude residue was purified by column chromatography on basic silica gel (100-200 mesh) using a hexane / EtOAc gradient (100 / 0 → 90 / 10 → 80 / 20 → 70 / 30 → 60 / 40 → 50 / 50) to give U as a yellow oil (0.7 g, 41%). 1H NMR (400 MHz, DMSO-d6): δ 7.8 (d, 1H), 7.24 (d, 1H), 4.49 (t, 1H), 4.36 (d, 2H), 3.26 (t, 2H), 2.88 (m, 2H), 1.7 (m, 3H), 1.14 (m, 2H), 0.26 (s, 9H). LCMS (ESI) 402.1 m / z [M+H] + .

[0625] Step-11: Synthesis of 6-(4-(((tert-butyldimethylsilyl)oxy)methyl)piperidin-1-yl)-2-nitro-3-(trimethylstannyl)pyridine (V) The title compound U (0.55 g, 1.37 mmol) and imidazole (1.3 g, 13.7 mmol) dissolved in DMF (11 mL) were cooled to 0 °C in an ice bath. DMAP (0.033 g, 0.27 mmol) and tert-butyldimethylsilyl chloride (1.0 g, 6.8 mmol) were added portionwise. The reaction mixture was stirred at room temperature for 2 h. The reaction mixture was quenched with water (100 mL) and extracted twice with ethyl acetate (100 mL). The organic phase was separated, dried over Na2SO4, filtered, and the solvent was removed under reduced pressure to give a crude residue. The crude residue was purified by column chromatography on basic silica gel (100-200 mesh) using a hexane / EtOAc gradient (100 / 0 → 99.5 / 0.5 → 99 / 1 → 98.5 / 1.5 → 98 / 2) to afford the title compound V as a yellow oil (0.32 g, 43%). 1 H NMR (400 MHz, DMSO-d6): δ 7.81 (d, 1H), 7.25 (d, 1H), 4.38 (d, 2H), 3.45 (d, 2H), 2.89 (t, 2H), 1.73 (d, 3H), 1.15 (m, 2H), 0.85 (s, 9H), 0.26 (s, 6H). LCMS (ESI) 514.7m / z [M] + .; 517.3 m / z [M+2H] + .

[0626] Step-12: 2-(6-(4-(((tert-butyldimethylsilyl)oxy)methyl)piperidin-1-yl)-2-nitropyridin-3-yl)-5-(pyridin-3-yl)-6,7-dihydrothiazolo[5,4-c]pyridin-4(5H)-one (W) Compound Q (0.3 g, 0.96 mmol) and compound V (0.74 g, 1.4 mmol) were dissolved in degassed 1,4-dioxane (45 mL). After the addition of Pd(OAc) (0.02 g, 0.1 mmol), XantPhos (0.055 g, 0.1 mmol), and Cu(I)Cl (0.095 g, 0.96 mmol), the reaction mixture was heated in an oil bath at 80 °C for 4 h. The reaction mixture was concentrated under reduced pressure to give a crude residue. The crude residue was purified by column chromatography on silica gel (100-200 mesh) using a CHCl / MeOH gradient (100 / 0 → 99 / 1 → 98 / 2) to give an impure solid. The impure solid was further purified by preparative TLC plate using dichloromethane / methanol (98 / 2) as the mobile phase to give compound W as a yellow solid (0.15 g, 26%). 1 H NMR (400 MHz, DMSO-d6): δ 8.65 (d, 1H), 8.45 (dd, 1H), 8.17 (d, 1H), 7.83 (m, 1H), 7.48 (dd, 1H), 7.23 (d, 1H), 4.39 (d, 2H), 4.13 LCMS (ESI) 581.5 m / z [M+H] + .

[0627] Step-13: 2-(6-(4-(hydroxymethyl)piperidin-1-yl)-2-nitropyridin-3-yl)-5-(pyridin-3-yl)-6,7-dihydrothiazolo[5,4-c]pyridin-4(5H)-one (20) Compound W (0.1 g, 0.17 mmol) was dissolved in THF (10 mL) and cooled to 0 °C with stirring in an ice bath. A 1 M solution of TBAF in THF (1 mL) was then added, and stirring was continued at RT for 2 h. The reaction was quenched with saturated aqueous sodium bicarbonate (pH 8–9) and extracted with 5% MeOH in DCM (2 × 25 mL). The organic phase was separated, dried over NaSO, filtered, and the solvent was removed under reduced pressure. The residue was purified by column chromatography on basic silica gel (100–200 mesh) using a CHCl / MeOH gradient (100 / 0 → 99 / 1 → 98 / 2) to give the desired compound containing some impurities. The impure material was further purified by preparative TLC using dichloromethane / ethanol (95 / 5) as the mobile phase to give the title compound 20 as a yellow solid (0.026 g, 34%). 1 H NMR (500 MHz, DMSO-d6): δ 8.65 (d, 1H), 8.45 (dd, 1H), 8.17 (d, 1H), 7.84-7.82 (m, 1H), 7.48 (dd, 1H), 7.23 (d, 1H), 4.53 (t, 1H), 4.38 (d, 2H), 4.14 (t, 2H), 3.28 (t, 2H), 3.19 (t, 2H), 3.00 (t, 2H), 1.78-1.67 (m, 3H). LCMS (ESI) 467.7 m / z [M+H] + .

[0628] Example 9 2-(2-fluoro-6-(4-(hydroxymethyl-t2)piperidin-1-yl)pyridin-3-yl)-5-(pyridin-3-yl)-6,7-dihydrothiazolo[5,4-c]pyridin-4(5H)-one ( 3 Synthesis of H-compound 17)

[0629] [ka]

[0630] T is 3 It means H.

[0631] A reaction vessel was charged with 150 μL n-BuLi (0.24 mmol, 1.6 M in hexane) and 50 μL TMEDA. The solution was degassed three times using a high-vacuum manifold and stirred at room temperature for 40 min under an atmosphere of tritium gas (1343 mbar, 9.2 Ci). This process was repeated twice (second time: 1097 mbar, 6.6 Ci and third time: 912 mbar, 4.8 Ci). At the end of the reaction (LiT formation), a pressure of 333 mbar was obtained. The solvent was removed under vacuum, and an AlCl solution (6.7 mg in 0.2 mL THF) was added with stirring. The mixture was stirred at room temperature for 10 min. A suspension of the precursor compound (3.3 mg in 0.5 mL THF) was added at −78 °C, and the reaction mixture was stirred at −78 °C for 3 h. The reaction vessel was cooled to −196 °C, and 0.2 mL HO was added. Due to the strong pressure buildup, warming to room temperature was carried out very slowly. The solvent was removed in vacuo, and the labile tritium was exchanged by adding 0.3 mL of methanol, stirring the solution, and again removing the solvent in vacuo. This process was repeated twice. Finally, the fully dried solid was dissolved in 5 mL of EtOH. The radiochemical purity of the crude material was determined to be approximately 44% using the following HPLC system: Waters Sunfire C18, 5 μm, 4.6 × 250 mm; Solvent A: water + 0.05% TFA, B: acetonitrile + 0.05% TFA; 0 min 20% B; 10–14.5 min 95% B, 15 min 20% B; 254 nm; 1.0 mL / min; 30 °C. The product had a retention time of 5.82 min. The following HPLC conditions were used for compound purification: Waters Sunfire C18, 10 x 250 mm; Solvent A: water + 0.1% TFA; B: acetonitrile + 0.1%); 27% B isocratic; 4.7 ml / min; 25 °C. The target compound eluted between 8.8 and 9.6 min. The solvent was removed in vacuo and the product was dissolved in 5 ml ethanol. The purified product had a specific activity of 55.4 Ci / mmol. 3 H-Compound 17, (97% purity) was obtained.

[0632] Example 10 Tritiated (R)-2-(2-fluoro-6-(3-fluoropyrrolidin-1-yl)pyridin-3-yl)-5-(pyridin-4-yl)-6,7-dihydrothiazolo[5,4-c]pyridin-4(5H)-one 3 H-Synthesis of compound 7

[0633] [ka]

[0634] T is 3 It means H.

[0635] 2.29 mg of brominated precursor 19 (described in Example 2 above) 3 Following the synthetic scheme for the [H] precursor, 50 μL DIPEA and 31.7 mg of Lindlar catalyst were suspended in 0.3 ml DMF. The suspension was degassed three times using a high-vacuum manifold and stirred overnight at room temperature under an atmosphere of tritium gas (401 mbar, 11.2 Ci). The solvent was removed in vacuo, and the labile tritium was exchanged by adding 0.3 ml of methanol, stirring the solution, and again removing the solvent in vacuo. This process was repeated twice. The thoroughly dried solid was extracted with 5 ml of ethanol / DMF (4:1), and the suspension was filtered through a 0.2 μm nylon membrane to obtain a clear solution. For the purification of the compound, the following HPLC conditions were found to be suitable: Waters Sunfire C18, 10 × 250 mm; Solvent A: water + 0.1% TFA; Solvent B: acetonitrile + 0.1% TFA. Purified product 3 H-Compound 7 (SA 50.8 Ci / mmol, 99% purity) was obtained.

[0636] Example 11 Tritiated (S)-2-(2-fluoro-6-(3-fluoropyrrolidin-1-yl)pyridin-3-yl)-5-(pyridin-3-yl)-6,7-dihydrothiazolo[5,4-c]pyridin-4(5H)-one ( 3Synthesis of H-compound 8)

[0637] [ka]

[0638] T is 3 It means H.

[0639] 2.45 mg of brominated precursor 20 (described in Example 2 above) 3 Following the synthetic scheme for the [H] precursor, 50 μL DIPEA and 33.5 mg of Lindlar catalyst were suspended in 0.3 ml DMF. The suspension was degassed three times using a high-vacuum manifold and stirred overnight at room temperature under an atmosphere of tritium gas (498 mbar, 4.3 Ci). The solvent was removed in vacuo, and the labile tritium was exchanged by adding 0.3 ml of methanol, stirring the solution, and again removing the solvent in vacuo. This process was repeated twice. The thoroughly dried solid was extracted with 5 ml of ethanol / DMF (4:1), and the suspension was filtered through a 0.2 μm nylon membrane to obtain a clear solution. For the purification of the compound, the following HPLC conditions were found to be suitable: Waters Sunfire C18, 10 × 250 mm; Solvent A: water + 0.1% TFA; Solvent B: acetonitrile + 0.1% TFA. Purified product 3 H-Compound 8 (SA 48.5 Ci / mmol, 99% purity) was obtained.

[0640] Example 12 Tritiated 2-(2-fluoro-6-(pyrrolidin-1-yl)pyridin-3-yl)-5-(pyridin-3-yl)-6,7-dihydrothiazolo[5,4-c]pyridin-4(5H)-one 3 Synthesis of H-compound 9

[0641] [ka]

[0642] T is 3 It means H.

[0643] 1.7 mg of brominated precursor 21 (described in Example 2 above) 3 Following the synthetic scheme for the [H] precursor, 20 μL DIPEA and 7.5 mg of Pd on carbon (10% metal) were suspended in 0.3 ml DMF. The suspension was degassed three times using a high-vacuum manifold and stirred overnight at room temperature under an atmosphere of tritium gas (670 mbar, 8.4 Ci). The solvent was removed in vacuo, and the labile tritium was exchanged by adding 0.3 ml of methanol, stirring the solution, and again removing the solvent in vacuo. This process was repeated twice. The thoroughly dried solid was extracted with 5 ml of ethanol / DMF (4:1), and the suspension was filtered through a 0.2 μm nylon membrane to obtain a clear solution. For the purification of the compound, the following HPLC conditions were found to be suitable: Waters Sunfire C18, 10 × 250 mm; Solvent A: water + 0.1% TFA; Solvent B: acetonitrile + 0.1% TFA. Purified product 3 H-Compound 9 (SA 57.6 Ci / mmol, 99% purity) was obtained.

[0644] Example 13 Tritiated 2-[2-fluoro-6-[(3R)-3-fluoropyrrolidin-1-yl]-3-pyridyl]-5-pyrimidin-2-yl-6,7-dihydrothiazolo[5,4-c]pyridin-4-one 3 Synthesis of H-compound 15

[0645] [ka]

[0646] T is 3 It means H.

[0647] Di-brominated precursor 22 (described in Example 2 above) 3Following the synthetic scheme for the [H] precursor, compound 15 (2.45 mg, user-supplied), DIPEA (50 μL, Acros A0408500), and Lindlar catalyst (15.5 mg) were suspended in DMF (0.4 ml, Acros 34831000). The suspension was degassed twice with a high vacuum manifold and stirred overnight at room temperature under an atmosphere of tritium gas (569 mbar, 7.7 Ci). At the end of the reaction, a pressure of 531 mbar was observed at room temperature. The solvent was removed in vacuo, and the labile tritium was exchanged by adding 0.3 ml of methanol, stirring the solution, and removing the solvent again in vacuo. This process was repeated twice. Finally, the thoroughly dried solid was extracted with 5 ml of DCM, and the suspension was filtered to obtain a clear solution. For the purification of the compound, the following HPLC conditions were found to be suitable: Waters Sunfire C18, 10 x 250 mm; Solvent A: Water (JT Baker HPLC gradient grade 4218) + 0.1% TFA (Sigma-Aldrich T6508); B: Acetonitrile (MACRON ChromAR HPLC ultra gradient 2856-25) + 0.1% TFA (Sigma-Aldrich T6508); 55% B isocratic; 4.7 ml / min; 25°C. Purified product 3 H-Compound 15 was obtained (SA 52 Ci / mmol, 99% purity).

[0648] Example 14 Fluorinated 2-(2-(fluoro- 18 Synthesis of F)-6-(4-(hydroxymethyl)piperidin-1-yl)pyridin-3-yl)-5-(pyridin-3-yl)-6,7-dihydrothiazolo[5,4-c]pyridin-4(5H)-one 18 F Compound 17

[0649] [ka]

[0650] [ in shipping vials (O-18 enriched water obtained from a commercial cyclotron facility) 18 [F] fluoride was transferred onto a Chromafix PS-HCO cartridge where it was collected (activated with KOTf (0.2 M, 6 mL) followed by water (6 mL)). 222 The solution was eluted into the reaction vessel of a TRACERlab® module with a solution of 15 / 3 mg of HCl / KCO (0.3 mL). The solution was first heated to 70°C for 3.5 min under vacuum and helium flow, and the thermal treatment was continued for an additional 2.5 min at 95°C under the same conditions. The reactor was then cooled to 60°C. A solution of 2-(6-(4-(hydroxymethyl)piperidin-1-yl)-2-nitropyridin-3-yl)-5-(pyridin-3-yl)-6,7-dihydrothiazolo[5,4-c]pyridin-4(5H)-one 20 (3.35 mg in 0.3 mL DMSO) was added to the reaction vial, and the vessel was heated to 160°C for 10 min. The reaction vessel was cooled to 60°C, and the mixture was diluted with HPLC buffer (4 mL). Purification was performed by HPLC using a semi-preparative Xbridge C18 column (5 μm, 250 × 10 mm) and elution with a mixture of acetonitrile / ammonium acetate solution (20 mM, 35 / 65, v / v) at a flow rate of 5 mL / min. The radiolabeled product was eluted from the SPE cartridge with 1.0 mL of ethanol into a formulation flask pre-filled with 10 mL of formulation base (10 mg of ascorbic acid in saline). The resulting solution was passed through a sterile 0.2 μm membrane filter into a sterile filter-vented vial. The final product 18 F - Compound 17 was assayed and a sample removed for QC testing. Confirmation of product identity was 19 F-Determined by co-injection of a sample of standard compound 17.

[0651] Synthesis scheme for (S)-2-(2-fluoro-6-(3-fluoropyrrolidin-1-yl)pyridin-3-yl)-5-(isothiazol-4-yl)-6,7-dihydrothiazolo[5,4-c]pyridin-4(5H)-one (Compound 25)

[0652] [ka]

[0653] Example 15 Synthesis of (S)-2-(2-fluoro-6-(3-fluoropyrrolidin-1-yl)pyridin-3-yl)-5-(isothiazol-4-yl)-6,7-dihydrothiazolo[5,4-c]pyridin-4(5H)-one

[0654] [ka]

[0655] Step-1: Synthesis of tert-butyl (S)-2-(2-fluoro-6-(3-fluoropyrrolidin-1-yl)pyridin-3-yl)-4-oxo-6,7-dihydrothiazolo[5,4-c]pyridine-5(4H)-carboxylate (G) The title compound F (0.26 g, 0.7 mmol), (S)-3-fluoropyrrolidine hydrochloride (0.088 g, 0.7 mmol), and N,N-diisopropylethylamine (0.2 mL, 1.4 mmol) were suspended in DMSO (26 mL) and stirred at RT for 2 h. After completion, the reaction mixture was suspended in water (260 mL), and the precipitated solid was filtered on Whatman filter paper. The solid was washed with water (20 mL) and dried under reduced pressure to give the title compound G as a pale yellow solid (0.27 g, 91%). 1 H NMR (500 MHz, DMSO-d6): δ 8.39 (t, 1H), 6.63 (d, 1H), 5.48 (d, 1H), 4.06 (t, 2H), 3.70 (dd, 3H), 3.54-3.49 (m, 1H), 3.10 (t, 2H), 2.30-2.27 (m, 2H), 1.49 (S, 9H).LCMS (ESI) 437.10 m / z [M+H] + .

[0656] Step-2: Synthesis of (S)-2-(2-fluoro-6-(3-fluoropyrrolidin-1-yl)pyridin-3-yl)-6,7-dihydrothiazolo[5,4-c]pyridin-4(5H)-one (H) The title compound G (0.27 g, 0.6 mmol) was dissolved in DCM (6 mL) and cooled to 0 °C with stirring in an ice bath. Then, 4 M HCl in 1,4-dioxane (2.7 mL) was added to the above solution, and stirring was continued at RT for 2 h. After completion of the reaction, the solvent was removed under reduced pressure. The resulting residue was dissolved in ice-cold water and basified to pH 8-9 with saturated aqueous sodium bicarbonate. The precipitated solid was filtered and dried to give the title compound H as a pale yellow solid (0.19 g, 91%). 1H NMR (500 MHz DMSO-d6): δ 8.39-8.35 (m, 1H), 7.82 (s, 1H), 6.61-6.60 (m, 1H), 5.48 (d, 1H), 3.80-3.63 (m, 4H), 3.53-3.48 (m, 2H), 2.99 (t, 2H), 2.30 - 2.18 (m, 2H). LCMS (ESI) 337 [M+H] + .

[0657] Step-3: Synthesis of (S)-2-(2-fluoro-6-(3-fluoropyrrolidin-1-yl)pyridin-3-yl)-5-(isothiazol-4-yl)-6,7-dihydrothiazolo[5,4-c]pyridin-4(5H)-one (1) Compound H (0.05 g, 0.14 mmol) and 4-bromoisothiazole (0.073 g, 0.44 mmol) were dissolved in degassed 1,4-dioxane (5 mL). After the addition of CuI (0.014 g, 0.074 mmol), 1,10-phenanthroline (0.026 g, 0.014 mmol), and CsF (0.045 g, 0.29 mmol), the reaction mixture was heated in a sand bath at approximately 100 °C for 48 h. The reaction mixture was concentrated under reduced pressure to give a crude residue. The crude residue was purified by column chromatography on silica gel (100-200 mesh) using a CHCl / MeOH gradient (100 / 0 → 99.5 / 0.5 → 99 / 1) to give pure compound 25 as a pale yellow solid (0.029 g, 46%). 1H NMR (400 MHz DMSO-d6): δ 8.95 (s, 1H), 8.88 (s, 1H), 8.43-8.38 (m, 1H), 6.65-6.62 (m, 1H), 5.48 (d, 1H), 4.22 (t, 2H), 3.85-3.72 (m, 3H), 3.55-3.48 (m, 1H), 3.27 (t, 2H), 2.27-2.16 (m, 2H). LCMS (ESI) 418.2 m / z [MH] +

[0658] Synthesis scheme for (S)-2-(2-fluoro-6-(3-fluoropiperidin-1-yl)pyridin-3-yl)-5-(isothiazol-4-yl)-6,7-dihydrothiazolo[5,4-c]pyridin-4(5H)-one (compound 27)

[0659] [ka]

[0660] Example 16 Synthesis of (S)-2-(2-fluoro-6-(3-fluoropiperidin-1-yl)pyridin-3-yl)-5-(isothiazol-4-yl)-6,7-dihydrothiazolo[5,4-c]pyridin-4(5H)-one

[0661] [ka]

[0662] Step-1: Synthesis of tert-butyl (S)-2-(2-fluoro-6-(3-fluoropiperidin-1-yl)pyridin-3-yl)-4-oxo-6,7-dihydrothiazolo[5,4-c]pyridine-5(4H)-carboxylate (G) The title compound F (0.4 g, 1.0 mmol), (S)-3-fluoropiperidine hydrochloride (0.15 g, 1.0 mmol), and N,N-diisopropylethylamine (0.36 mL, 2.1 mmol) were suspended in DMSO (40 mL), and the reaction mixture was stirred at RT for 2 h. After completion, the reaction mixture was suspended in water (400 mL) and filtered through a Whatman filter paper. The solid was washed with water (20 mL) and dried under reduced pressure to give the title compound G as a pale yellow solid (0.47 g, 96%). 1 H NMR (500 MHz, DMSO-d6): δ 8.37-8.33 (m, 1H), 6.97-6.95 (m, 1H), 4.85 (d, 1H), 4.22-4.16 (m, 1H), 4.06 (t, 2H), 4.01 (d,1H), 3.67-3.57 (m, 1H), 3.38-3.35 (m, 1H), 3.10 (t, 2H), 1.95-1.87 (m, 2H), 1.78-1.70 (m, 1H), 1.61-1.56 (m, 1H), 1.49 (s, 9H), LCMS (ESI) 451.35 m / z [M+H] + .

[0663] Step-2: Synthesis of (S)-2-(2-fluoro-6-(3-fluoropiperidin-1-yl)pyridin-3-yl)-6,7-dihydrothiazolo[5,4-c]pyridin-4(5H)-one (H) The title compound G (0.47 g, 1.04 mmol) was dissolved in DCM (9 mL) and cooled to 0 °C with stirring in an ice bath. 4 M HCl in 1,4-dioxane (4.7 mL) was added to the above solution, and stirring was continued at RT for 2 h. After completion of the reaction, the solvent was removed under reduced pressure. The resulting residue was dissolved in ice-cold water and basified to pH 8-9 with saturated aqueous sodium bicarbonate. The precipitated solid was filtered and dried to give the title compound H as a pale yellow solid (0.36 g, 98%). 1H NMR (500 MHz DMSO-d6): δ 8.35-8.31 (m, 1H), 7.82 (s, 1H), 6.95-6.93 (m, 1H), 4.90-4.79 (m, 1H), 4.18-4.13 (m, 1H), 3.99-3.96 (m, 1H), 3.66-3.60 (m, 1H), 3.58-3.49 (m, 2H), 3.36-3.34 (m, 1H), 2.99 (t, 2H), 1.94-1.87 (m, 2H), 1.74-1.73 (m, 1H), 1.60-1.56 (m, 1H). LCMS (ESI) 351 [M+H] + .

[0664] Step-3: Synthesis of (S)-2-(2-fluoro-6-(3-fluoropiperidin-1-yl)pyridin-3-yl)-5-(isothiazol-4-yl)-6,7-dihydrothiazolo[5,4-c]pyridin-4(5H)-one (28) Compound H (0.05 g, 0.14 mmol) and 4-bromoisothiazole (0.07 g, 0.42 mmol) were dissolved in degassed 1,4-dioxane (5 mL). After the addition of CuI (0.013 g, 0.07 mmol), 1,10-phenanthroline (0.025 g, 0.14 mmol), and CsF (0.043 g, 0.28 mmol), the reaction mixture was heated in a sand bath at approximately 100 °C for 48 h. The reaction mixture was concentrated under reduced pressure to give the crude product. The crude residue was purified by column chromatography on silica gel (100-200 mesh) using a CHCl / MeOH gradient (100 / 0 → 99.5 / 0.5 → 99 / 1 → 98.5 / 1.5) to give pure compound 27 as a yellow solid (0.047 g, 77%). 1H NMR (400 MHz DMSO-d6): δ 8.95 (s, 1H), 8.88 (s, 1H), 8.39-8.35 (m, 1H), 6.99-6.96 (m, 1H), 4.92-4.79 (m, 1H), 4.23-4.16 (m, 3H), 4.03-4.00 (m, 1H), 3.68-3.57 (m, 1H), 3.37-3.36 (m, 1H), 3.27 (t, 2H),1.94-1.87 (m, 2H), 1.76-1.58 (m, 2H). LCMS (ESI) 433.9 m / z [MH] +

[0665] Synthesis scheme: (R)-2-(2-fluoro-6-(3-fluoropiperidin-1-yl)pyridin-3-yl)-5-(isothiazol-4-yl)-6,7-dihydrothiazolo[5,4-c]pyridin-4(5H)-one (compound 24)

[0666] [ka]

[0667] Example 17 Synthesis of (R)-2-(2-fluoro-6-(3-fluoropiperidin-1-yl)pyridin-3-yl)-5-(isothiazol-4-yl)-6,7-dihydrothiazolo[5,4-c]pyridin-4(5H)-one

[0668] [ka]

[0669] Step-1: Synthesis of tert-butyl (R)-2-(2-fluoro-6-(3-fluoropiperidin-1-yl)pyridin-3-yl)-4-oxo-6,7-dihydrothiazolo[5,4-c]pyridine-5(4H)-carboxylate (G) The title compound F (0.6 g, 1.63 mmol), (R)-3-fluoropiperidine hydrochloride (0.22 g, 1.63 mmol), and N,N-diisopropylethylamine (0.55 mL, 3.27 mmol) were suspended in DMSO (60 mL) and stirring was continued at RT for 2 h. After completion of the reaction, the reaction mixture was suspended in water (600 mL) and filtered on Whatman filter paper. The solid was washed with water (20 mL) and dried under reduced pressure to give the title compound G as a pale yellow solid (0.63 g, 90%). 1 H NMR (500 MHz, DMSO-d6): δ 8.35 (t, 1H), 6.97-6.95 (m, 1H), 4.85 (d, 1H), 4.21-4.16 (m, 1H), 4.06-3.99 (m, 3H), 3.67-3.57 (m, 1H), 3.38-3.34 (m, 1H), 3.11-3.08 (t, 2H), 1.93-1.87 (m, 2H), 1.76-1.73 (m, 1H), 1.61-1.56 (m, 1H), 1.49 (s, 9H), LCMS (ESI) 451.25 m / z [M+H] + .

[0670] Step-2: Synthesis of (R)-2-(2-fluoro-6-(3-fluoropiperidin-1-yl)pyridin-3-yl)-6,7-dihydrothiazolo[5,4-c]pyridin-4(5H)-one (H) The title compound G (0.63 g, 1.4 mmol) was dissolved in DCM (12 mL) and cooled to 0 °C with stirring in an ice bath. 4 M HCl in 1,4-dioxane (6.3 mL) was added to the above solution, and stirring was continued at RT for 2 h. After completion of the reaction, the solvent was removed under reduced pressure. The resulting residue was dissolved in ice-cold water and basified with saturated aqueous sodium bicarbonate to pH 8-9. The precipitated solid was filtered and dried to give the title compound H as a pale yellow solid (0.4 g, 85%). 1H NMR (500 MHz DMSO-d6):δ 8.36-8.32 (m, 1H), 7.83 (s, 1H), 6.96-6.94 (m, 1H), 4.89-4.79 (m, 1H), 4.18-4.13 (m, 1H), 3.99-3.96 (m, 1H), 3.66-3.57 (m, 1H), 3.51-3.48 (m, 2H), 3.36-3.34 (m, 1H), 2.99 (t, 2H), 1.95-1.87 (m, 2H), 1.77-1.71 (m, 1H), 1.60-1.56 (m, 1H). LCMS (ESI) 351 [M+H] + .

[0671] Step-3: Synthesis of (R)-2-(2-fluoro-6-(3-fluoropiperidin-1-yl)pyridin-3-yl)-5-(isothiazol-4-yl)-6,7-dihydrothiazolo[5,4-c]pyridin-4(5H)-one (24) Compound H (0.05 g, 0.14 mmol) and 4-bromoisothiazole (0.07 g, 0.42 mmol) were dissolved in degassed 1,4-dioxane (5 mL). After the addition of CuI (0.013 g, 0.07 mmol), 1,10-phenanthroline (0.025 g, 0.14 mmol), and CsF (0.043 g, 0.28 mmol), the reaction mixture was heated in a sand bath at approximately 100 °C for 36 h. The reaction mixture was concentrated under reduced pressure to give the crude product. The crude product thus obtained was purified by column chromatography on silica gel (100-200 mesh) using a CHCl / MeOH gradient (100 / 0 → 99.5 / 0.5 → 99 / 1) to give pure compound 24 as a yellow solid (0.03 g, 50%). 1H NMR (400 MHz DMSO-d6): δ 8.95 (s, 1H), 8.88 (s, 1H), 8.39-8.35 (m, 1H), 6.99-6.96 (m, 1H), 4.91-4.79 (m, 1H), 4.23-3.99 (m, 4H), 3.68-3.57 (m, 1H), 3.40-3.39 (m, 1H), 3.28-3.24 (m, 2H),1.96-1.87 (m, 2H), 1.76-1.57 (m, 2H). LCMS (ESI) 433.8 m / z [MH] +

[0672] Synthesis scheme for 2-(2-fluoro-6-(piperidin-1-yl)pyridin-3-yl)-5-(2-fluoropyridin-4-yl)-6,7-dihydrothiazolo[5,4-c]pyridin-4(5H)-one (compound 23)

[0673] [ka]

[0674] Example 18 Synthesis of 2-(2-fluoro-6-(piperidin-1-yl)pyridin-3-yl)-5-(2-fluoropyridin-4-yl)-6,7-dihydrothiazolo[5,4-c]pyridin-4(5H)-one

[0675] [ka]

[0676] Step-1: Synthesis of tert-butyl 2-(2-fluoro-6-(piperidin-1-yl)pyridin-3-yl)-4-oxo-6,7-dihydrothiazolo[5,4-c]pyridine-5(4H)-carboxylate (G) The title compound F (0.3 g, 0.81 mmol), piperidine (0.069 g (0.08 mL), 0.81 mmol), and N,N-diisopropylethylamine (0.2 mL, 1.6 mmol) were suspended in DMSO (30 mL) and stirred at RT for 2 h. After completion, the reaction mixture was suspended in water (300 mL) and filtered over Whatman filter paper. The solid was washed with water (20 mL) and dried under reduced pressure to give the title compound G as a pale yellow solid (0.34 g, 98%). 1 H NMR (500 MHz, DMSO-d6): δ 8.32 (t, 1H), 6.91-6.89 (m, 1H), 4.05 (t, 2H), 3.65 (t, 4H), 3.09 (t, 2H), 1.64 (t, 2H), 1.58-1.53 ​​(m, 4H), 1.48 (s, 9H). LCMS (ESI) 432.90 m / z [M+H] + .

[0677] Step-2: Synthesis of 2-(2-fluoro-6-(piperidin-1-yl)pyridin-3-yl)-6,7-dihydrothiazolo[5,4-c]pyridin-4(5H)-one (H) The title compound G (0.34 g, 0.78 mmol) was dissolved in DCM (7 mL) and cooled to 0 °C with stirring in an ice bath. 4 M HCl in 1,4-dioxane (3.4 mL) was added to the above solution, and stirring was continued at RT for 2 h. After completion of the reaction, the solvent was removed under reduced pressure. The resulting residue was dissolved in ice-cold water and basified with saturated aqueous sodium bicarbonate to pH 8-9. The precipitated solid was filtered and dried to give the title compound H as a yellow solid (0.255 g, 98%). 1H NMR (400 MHz DMSO-d6): δ 8.34-8.29 (m, 1H), 7.83 (s, 1H), 6.90-6.87 (m, 1H), 3.65-3.62 (m, 4H), 3.51-3.47 (m, 2H), 2.99 (t, 2H), 1.65-1.63 (m, 2H), 1.56-1.55 (m, 4H). LCMS (ESI) 333.2 [M+H] + .

[0678] Step-3: Synthesis of 2-(2-fluoro-6-(piperidin-1-yl)pyridin-3-yl)-5-(2-fluoropyridin-4-yl)-6,7-dihydrothiazolo[5,4-c]pyridin-4(5H)-one (4) Compound H (0.05 g, 0.15 mmol) and 4-bromo-2-fluoropyridine (0.039 g, 0.22 mmol) were dissolved in degassed 1,4-dioxane (5 mL). After the addition of Pd[PPh3]4 (0.008 g, 0.007 mmol), XantPhos (0.008 g, 0.015 mmol), and Cs2CO3 (0.073 g, 0.22 mmol), the reaction mixture was heated in a sand bath at approximately 100 °C for 24 h. The reaction mixture was concentrated under reduced pressure to give a crude residue. The residue was purified by column chromatography on silica gel (100-200 mesh) using a CHCl2 / MeOH gradient (100 / 0 → 99.5 / 0.5 → 99 / 1) to give the impure compound. The impure compound was further purified by preparative TLC plate using CH2Cl2 / MeOH (98.5 / 1.5) as the mobile phase to give the title compound 23 as a pale yellow solid (0.032 g, 50%). 1 H NMR (400 MHz DMSO-d6): δ 8.37-8.32 (m, 1H), 8.22 (d, 1H), 7.47-7.45 (m, 1H), 7.25 (d, 1H), 6.93-6.90 (m, 1H), 4.22 (t, 2H), 3.66 (t, 4H), 3.24 (t, 2H), 1.65-1.63 (m, 2H), 1.57-1.56 (m, 4H). LCMS (ESI) 427.9 m / z [M+H] +

[0679] Biological assay description 1. General method Human brain material for these studies was obtained from the Neurodegenerative Disease Brain Bank, UCSF, Prof. William Seeley (funded by NIH grants P01AG019724 and P50AG023501, the Consortium for Frontotemporal Dementia Research, and the Tau Consortium) and from the Queen Square Brain Bank for Neurological Disorders, UCL, Prof. Tammaryn Lashley. All material was collected from donors whose written informed consent was obtained by the brain bank for brain autopsy and the use of their material and clinical information for research purposes.

[0680] 1.1 Radioligands It has a specific activity of 48.5 Ci / mmol (1.0 mCi / mL) as described above [ 3 H]-Compound 1, described above, with a specific activity of 50.8 Ci / mmol (1.0 mCi / mL) [ 3 H]-Compound 7, described above, with a specific activity of 55.1 Ci / mmol (1.0 mCi / mL) [ 3 H]-Compound 8, described above, with a specific activity of 57.6 Ci / mmol (1.0 mCi / mL) [ 3 H]-Compound 9, described above, with a specific activity of 53.3 Ci / mmol (1.0 mCi / mL) [ 3 H]-Compound 15, described above, with a specific activity of 52 Ci / mmol (1.0 mCi / mL) [ 3 H]-Compound 16, described above, with a specific activity of 55.4 Ci / mmol (1.0 mCi / mL) [ 3 H]-compound 17, and / or [ 3 H]-standard compounds (see WO2023 / 285661 for [ 3 H]-a compound described as Compound 16) is used in the following assays.

[0681] 1.2. Preparation of human frontotemporal dementia (FTD) sarkosyl-insoluble brain extracts Human brain extracts were prepared as described in Laferriere et al., 2019, Nature Neurosc. Brain tissue samples (frontal or temporal cortex) were homogenized at 4°C in a 1:4 (w / v) ratio in homogenization-solubilization (HS) buffer using a tissue homogenizer (Precellys) with a CKmix homogenization tube. The following sequence was used for homogenization: 5000 rpm for 30 seconds, three cycles with a 15-second pause between each cycle. Homogenized samples were aliquoted and stored at -80°C in 1.5 mL low-protein binding tubes.

[0682] Brain homogenates were thawed on ice and resuspended in HS buffer to a final concentration of 2% sarkosyl, 1 unit / μL Benzonase, and 1 mM MgCl2. The samples were then incubated at 37°C for 45 minutes (min) under constant shaking at 600 rpm on a thermomixer. The supernatant was collected in a new tube (sarkosyl-soluble fraction, S1). The pellet was resuspended in 1000 μL of myelin suspension buffer and centrifuged at 20,000 g for 60 min at 4°C. The supernatant was carefully removed to remove all floating lipids. This step was repeated if all lipids could not be removed in a single step. The pellet was then washed with phosphate-buffered saline (PBS) and centrifuged at 20,000 g for 30 min at 4°C. The final pellet was resuspended in 200 μL PBS and stored at -80°C (sarkosyl-insoluble fraction). Samples were analyzed by immunoblotting under denaturing conditions.

[0683] 2. Description of the biological assay and corresponding results 2.1. Microradiobinding competition assay to determine binding affinity to TPD-43 Human FTD sarkosyl-insoluble brain extracts were spotted onto microarray slides. Slides were incubated with 40 nM tritiated reference ligand and 2 μM and 250 nM (non-radiolabeled) example compounds. In some cases, non-radiolabeled example compounds were further evaluated at a series of different concentrations ranging from 0.24 nM to 2 μM to determine the inhibition constant (Ki). After incubation, slides were washed and scanned using a real-time autoradiography system (BeaQuant, ai4R). Signal quantification was performed using Beamage image analysis software (ai4R). Nonspecific signal was determined with an excess of non-radiolabeled standard compound (2 μM), and specific binding was calculated by subtracting the nonspecific signal from the total signal. Competition was calculated as a percentage, with 0% defined as specific binding in the presence of vehicle and 100% defined as the value obtained in the presence of an excess of non-radiolabeled standard compound. Calculate K in GraphPad Prism 8 by applying a nonlinear regression curve fit using a one-site specific binding model. i Values ​​were calculated. Measurements were performed with at least two technical replicates in two-concentration competition experiments and one technical replicate in experiments involving a range of concentrations. For compounds tested in more than one experiment, replicate values ​​or K values ​​from independent experiments were used. i The average values ​​are reported.

[0684] Results: Example compounds were tested for their ability to bind to TDP-43 aggregates derived from the brains of FTD patients. 3 The compounds were evaluated for their potency in competing with the binding of a [H]-reference ligand. The results of the microradio binding competition assay with the example compounds are shown in Table 5 below: % competition at 2 μM and 250 nM. i The values ​​are also shown in Table 5.

[0685] [Table 6A]

[0686] [Table 6B]

[0687] [Table 6C]

[0688] [Table 6D]

[0689] 2.2 [ 3 H]-Kd determination of compounds 1, 7, 8, 9, 15, 16, and 17 in human FTD sarkosyl-insoluble brain extracts in a microradio binding assay Human FTD sarkosyl-insoluble brain extract was spotted onto microarray slides. 3 The slides were incubated with [H]-compounds 1, 7, 8, 9, 15, 16, and 17 at a series of concentrations ranging from 1.29 to 300 nM. After incubation, the slides were washed and scanned using a real-time autoradiography system (BeaQuant, ai4R). Signal quantification was performed using Beamage image analysis software (ai4R). Nonspecific signals were determined with an excess of non-radiolabeled compounds 1, 7, 8, 9, 15, 16, and 17 (1 μM), respectively, and specific binding was calculated by subtracting the nonspecific signal from the total signal.

[0690] Kd (dissociation constant) and R 2 (a parameter ranging from 0.0 to 1.0 that quantifies the goodness of fit, with the best curve fit being obtained at a value of 1.0) was obtained by fitting the specific binding data with nonlinear regression analysis using a one-site specific binding model in GraphPad Prism 8.

[0691] result: [ 3The dissociation constants (Kd) of [H]-compounds 1, 7, 8, 9, 15, 16, and 17 were determined in a microradiobinding assay with human FTD type A sarkosyl-insoluble brain extracts. 3 H]-Compounds 1, 7, 8, 9, 15, 16, and 17 had high specific binding and produced a high dynamic range. 3 [H]-Compound 1 exhibited a Kd value of 69 nM in human FTD sarkosyl-insoluble brain extracts (Fig. 1a). Data from two independent experiments yielded an average Kd of 69 ± 21 nM. 3 [H]-Compound 7 exhibited a Kd value of 39 nM in human FTD sarkosyl-insoluble brain extracts (Fig. 1b). Data from three independent experiments yielded an average Kd of 39 ± 11 nM. 3 [H]-Compound 8 exhibited a Kd value of 47 nM in human FTD sarkosyl-insoluble brain extracts (Fig. 1c). Data from two independent experiments yielded an average Kd of 47 ± 8 nM. 3 H]-Compound 9 showed a Kd value of 26 nM in human FTD sarkosyl-insoluble brain extracts (Fig. 1d). 3 H]-Compound 15 exhibited a Kd value of 21 ± 7 nM in human FTD sarkosyl-insoluble brain extracts (Fig. 1e). 3 H]-Compound 16 exhibited a Kd value of 24 ± 3 nM in human FTD sarkosyl-insoluble brain extracts (Fig. 1f). 3 H]-Compound 17 exhibited a Kd value of 25±6 nM in human FTD sarkosyl-insoluble brain extracts (Fig. 1g). 3 H]-Compound 1 and [ 3 The dissociation constant (Kd) for [H]-compound 17 was also determined in a microradio binding assay with human FTD type B sarkosyl-insoluble brain extracts. 3 H]-Compound 1 showed a Kd value of 49 nM in human FTD type B sarkosyl-insoluble brain extracts (Fig. 1h), and [ 3 H]-Compound 17 showed a Kd value of 12 nM (Fig. 1i).

[0692] 2.3. In brain sections from patients with FTD 3 H]-Compound 1, [ 3 H]-compound 9 and [3 Micro-autoradiography staining with [H]-compound 17 All tissues were collected from donors, whose written informed consent for brain autopsy and the use of their materials and clinical information for research purposes was obtained from or by the respective brain banks. All samples were anonymized and coded. Frozen brain tissue blocks with confirmed TDP-43 pathology at autopsy were processed using a cryotome to generate 10-μm-thick sections, which were then mounted on glass slides. Sections were stored at -80°C until use.

[0693] Brain sections were immunostained using a commercially available antibody specific for phosphorylated serine at amino acids 409 / 410 (anti-pTDP-43 pS409 / 410, Biolegend, 829901). Sections were fixed with 4% formaldehyde (Sigma, 252549) for 15 min at 4°C and washed three times with 1x PBS (Dulbecco's Phosphate-Buffered Saline, Sigma, D1408) for 8 min each at room temperature. Next, sections were saturated and permeabilized in blocking buffer (PBS, 10% normal goat serum (NGS), 0.25% Triton X-100) for 1 h at room temperature and then incubated overnight at 4°C with primary antibody against pTDP-43 (diluted 1 / 500 in PBS, 5% NGS, 0.25% Triton X-100). The next day, sections were washed three times for 8 min with 1x PBS and then incubated with secondary AlexaFluor633-conjugated goat anti-rat antibody (Invitrogen, A-21094, diluted 1 / 500 in PBS) for 45 min at room temperature. After incubation with the primary antibody, sections were washed three times in PBS before further processing.

[0694] For micro-autoradiography, tritiated test compounds (respectively [ 3 H]-Compound 1, [ 3 H]-compound 9 or [ 3[H]-Compound 17) was applied to the sections and incubated at 120 nM in 50 mM Tris buffer, pH 7.4, for 45 minutes at room temperature. The sections were then washed as follows: once for 1 minute in ice-cold 50 mM Tris-HCl buffer, pH 7.4, twice for 1 minute in ice-cold 70% ethanol, once for 1 minute in ice-cold 50 mM Tris-HCl buffer, pH 7.4, and finally briefly rinsed in ice-cold distilled water. The sections were then dried under airflow for 1 hour and then exposed to Ilford Nuclear Emulsion Type K5 (Agar Scientific, AGP9281) in a light-tight slide storage box at 4°C for 10 days. Sections were developed by successively immersing them in the following solutions: 1.) Ilford Phenisol Developer (diluted 1:5 in H2O, Agar Scientific, AGP9106) for 4 min, 2.) Ilfostop solution (diluted 1:20 in H2O, Agar Scientific, AGP9104) for 2 min, 3.) Ilford Hypam Fixer (diluted 1:5 in H2O, Agar Scientific, AGP9183) for 4 min, and finally rinsed in H2O for 10 min.

[0695] For image acquisition, sections were mounted and imaged using ProLong Gold Antifade reagent (Invitrogen P36930) on a Panoramic 250 slide scanner (3DHistech) with a 40x objective, which captured bright-field and fluorescent images separately, or on a Panoramic Scan II (3DHistech) with a 20x objective. Fluorescent and bright-field images were aligned using the Visiopharm image analysis software suite.

[0696] result: Tritiated test compounds ([ 3 H]-Compound 1, [ 3 H]-Compound 9, [ 3The micro-autoradiography signal from [H]-compound 17) was detected in the form of accumulated silver granules that colocalized with the immunofluorescence signal from the pTDP-43-specific antibody, thereby confirming the [H]-specific antibody binding to TDP-43 aggregates. 3 H]-Compound 1, [ 3 H]-compound 9 and [ 3 This suggests target engagement of [H]-compound 17. 3 H]-Compound 1 (120 nM) (Figure 2a), [ 3 H]-compound 9 (Fig. 2b) and [ 3 Further evidence of target engagement was provided by incubation with [H]-compound 17 (Figure 2c), which showed colocalization with pTDP-43 aggregates. The colocalization was also observed in [H]-compound 17 on human brain sections from FTLD-TDP type B donors. 3 H]-Compound 1 and [ 3 H]-compound 17 (Figures 2d and 2e, respectively).

[0697] 2.4 In brain sections from patients with FTD, [ 3 H]-Compound 1, [ 3 H]-compound 8 and [ 3 Autoradiography using [H]-compound 17 All tissues were collected from donors, whose written informed consent for brain autopsy and the use of their materials and clinical information for research purposes was obtained from or by the respective brain banks. All samples were anonymized and coded. Frozen brain tissue blocks with confirmed TDP-43 pathology at autopsy were processed using a cryotome to generate 10-μm-thick sections, which were then mounted on glass slides. Sections were stored at -80°C until use.

[0698] Brain sections were fixed in 4% formaldehyde on ice for 15 minutes at 4°C and washed three times with 1x PBS for 8 minutes at room temperature. Sections were blocked for 30 minutes in assay buffer (50 mM Tris-HCl, 0.9% NaCl + 0.1% BSA). Cold test compound (Compound 1, Compound 8, or Compound 17) was diluted to 4 μM in assay buffer and applied to half of the sections for 40 minutes at room temperature to determine nonspecific binding. Only the other half of the sections was incubated with assay buffer (total binding). After this incubation, an equal volume of the corresponding tritiated compound (i.e., [ 3 H]-Compound 1, [ 3 H]-compound 8 or [ 3 H]-compound 17) at 20 nM ([ 3 H]-Compound 1 and [ 3 H]-compound 8) or 10 nM ([ 3 H]-compound 17) or at concentrations ranging from 0.1 to 200 nM ([ 3 H]-Compound 1 and [ 3 H]-compound 17), added to the sections, and incubated for 2 h at RT, followed by washing as follows: once for 1 min in ice-cold 50 mM Tris-HCl buffer, pH 7.4, twice for 1 min in ice-cold PBS, once for 1 min in ice-cold 50 mM Tris-HCl buffer, pH 7.4, and finally a brief rinse in ice-cold distilled water.

[0699] The sections were then dried under airflow and mounted with copper-based adhesive strips. Bound radioactivity was quantified by exposing the slides for 2 hours in a real-time autoradiography system (Beaquant™, AI4R) with the measurement settings set for tritium. Images were acquired using Beavacq software (AI4R) and subsequently quantified using Beamage image analysis software (AI4R). Specific binding (total binding minus nonspecific binding) and displacement (total binding divided by nonspecific binding) were calculated. Bar graphs were generated using GraphPad Prism. To determine Kd in brain sections, specific binding data were fitted to nonlinear regression analysis using a one-site specific binding model in GraphPad Prism.

[0700] result: Example compounds were evaluated for their binding on human tissue sections from non-demented control cases, FTLD-TDP type A, or FTLD-TDP type B (Figures 3a and 4a). Displacement of tritiated compounds by non-radiolabeled compounds was calculated in gray matter regions (total binding divided by non-specific binding) and is reported for each case in Table 6 below.

[0701] [Table 7]

[0702] Compounds 1, 8, and 17 showed increased substitution in FTLD-TDP types A and B compared to substitution in non-demented controls, providing further evidence of target engagement in pathological tissues. As can be seen in Figures 3b and 4b, 3 H]-Compound 1 and [ 3 [H]-Compound 17 exhibited significantly higher specific binding in FTLD-TDP type A compared to the control (large solid circles correspond to images of type A and type B shown in Figures 3a and 4a). One-way ANOVA with Tukey's correction for multiple comparisons. * p<0.05, ** p<0.01). 3 H]-Compound 1 and [ 3 [H]-Compound 17 was also evaluated in saturation binding studies using FTLD-TDP type A tissue sections, measuring Kd values ​​of 16 nM and 18 nM, respectively, suggesting that it binds to TDP-43 aggregates with high affinity (Figures 3c and 4c).

[0703] 2.5 Selectivity for TDP-43 2.5.1 Radiobinding assay in Alzheimer's disease (AD) brain homogenates 2.5.1.1 Preparation of Human AD Insoluble Fraction from Brain Homogenate The procedure used was adapted from Bagchi et al., 2013, which describes the extraction of a homogenized insoluble fraction containing protein aggregates from human brain tissue for in vitro binding and competition studies.

[0704] Approximately 4 g of frozen tissue blocks from the frontal cortex brain region of AD donors with confirmed tau and Abeta aggregate burden were used. The tissue was homogenized in three volumes (1:3 w / v) of high-salt buffer (50 mM Tris-HCl, pH 7.5, 750 mM NaCl, 5 mM EDTA) supplemented with protease inhibitors (Complete; Roche 11697498001) using a glass Dounce homogenizer at 4°C. The homogenate was transferred to polycarbonate centrifuge bottles (16 x 76 mm; Beckman 355603) and centrifuged at 100,000 x g (38,000 RPM) in a pre-cooled 70.1 rotor (Beckman 342184) in an ultracentrifuge (Beckman XL100K) for 60 minutes at 4°C. The pellet was resuspended in high-salt buffer supplemented with 1% Triton X-100 and homogenized with a syringe at 4°C. The homogenate was centrifuged again at 100,000 × g (38,000 RPM, 70.1 rotor) for 60 minutes at 4°C. The pellet was resuspended in high-salt buffer supplemented with 1% Triton X-100 and 1 M sucrose and homogenized with a syringe at 4°C. The homogenate was centrifuged at 100,000 × g (38,000 RPM, 70.1 rotor) for 60 minutes at 4°C. The resulting pellet, containing the insoluble fraction, was resuspended in PBS (1:2 w / v relative to the starting tissue mass), aliquoted, and stored at -80°C until use.

[0705] 2.5.1.2 Radiobinding Competition Assay Using Abeta Standard Compounds to Determine Inhibitor Constants (Ki) in the AD-Insoluble Fraction of Brain Homogenates A fixed concentration of AD insoluble fraction was incubated with a fixed concentration of radiolabeled Abeta standard compound (Kd = 33 nM in AD brain homogenate) and increasing concentrations of non-radiolabeled compound ranging from 0.41 nM to 2 μM. Reactions were performed in assay buffer (50 mM Tris, pH 7.5, 0.1% BSA in 0.9% NaCl) and incubated for 2 hours at room temperature (RT). Duplicate samples were then filtered under vacuum onto GF / C filter plates (PerkinElmer) to capture aggregates with bound radioligand and washed with 100 μL ice-cold 50 mM Tris, pH 7.5. The GF / C filters were then dried, and scintillation fluid (UltimateGold, PerkinElmer, 6013151) was added to each well. Filters were analyzed on a Microbeta2 scintillation counter (PerkinElmer). Values ​​for maximum signal were obtained without non-radiolabeled compound, while 100% displacement was obtained using 1 μM non-radiolabeled standard compound. K values ​​were calculated by non-linear regression using a one-site-fit K model in Prism V7 (GraphPad). The average K for non-radiolabeled Abeta standard compound is 32 nM.

[0706] 2.5.1.3 Radiobinding Assay to Determine Dissociation Constants (Kd) in the AD-Insoluble Fraction of Brain Homogenates The AD insoluble fraction was analyzed by different concentrations of [ 3 H] Compound 1, [ 3 H] Compound 17 or [ 3 H]A beta standard compound ([ 3The wells were incubated with [H]Abeta (ref). To determine nonspecific signal, 1 μM of non-radiolabeled compound was added to all concentrations of tritiated compound in control wells. Reactions were performed in assay buffer (50 mM Tris, pH 7.5, 0.1% BSA in 0.9% NaCl) and incubated for 2 hours at RT. Samples were then filtered under vacuum through GF / C filter plates (PerkinElmer) to capture / immobilize aggregates with bound radioligand and washed five times with 50 mM Tris, pH 7.5. The GF / C filters were then dried, and scintillation fluid (UltimateGold, PerkinElmer, 6013151) was added to each well. Filters were analyzed in a Microbeta scintillation counter (PerkinElmer). Specific binding was calculated by subtracting the nonspecific signal from the total signal. Kd and R 2 Values ​​were obtained by fitting the specific binding data to nonlinear regression analysis using a one-site specific binding model in GraphPad Prism.

[0707] 2.5.2. Microradiobinding assays in a-syn aggregates from PD brains and tau PHFs from AD cases 2.5.2.1 Isolation of pathological a-syn aggregates from PD brains The procedure was adapted from the protocol described in Spillantini et al., 1998. Frozen tissue blocks from PD donors were thawed on ice and homogenized using a glass Dounce homogenizer. The homogenate was then centrifuged at 11,000 × g (12,700 RPM) for 20 minutes at 4°C in a Beckman XL100K ultracentrifuge using a pre-chilled 70.1 rotor (Beckman, 342184). The pellet was resuspended in extraction buffer [10 mM Tris-HCl, pH 7.4, 10% sucrose, 0.85 mM NaCl, 1% protease inhibitor (Calbiochem 539131), 1 mM EGTA, 1% phosphatase inhibitor (Sigma P5726 and P0044)] and centrifuged at 15,000 × g (14,800 RPM, 70.1 Ti rotor) for 20 min at 4 °C. The pellet was discarded, and sarkosyl (20% stock solution, Sigma L7414) was added to the supernatant to a final concentration of 1% at room temperature for 1 h. This solution was then centrifuged at 100,000 × g (38,000 RPM, 70.1 Ti rotor) for 1 h at 4 °C. The pellet, containing enriched a-syn aggregates, was resuspended in PBS and stored at -80 °C until use.

[0708] 2.5.2.2 Preparation and Extraction of Paired Helical Filament (PHF)-Tau Aggregates from AD Brains The enrichment procedure was modified from Jicha et al., 1997, and Rostagno and Ghiso, 2009. Briefly, approximately 15 g of AD human brain was thawed on ice and homogenized in a glass Dounce homogenizer in homogenization buffer (3 ml per gram of tissue) (0.75 M NaCl in RAB buffer (100 mM 2-(N-morpholino)ethanesulfonic acid (MES), 1 mM EGTA, 0.5 mM MgSO4, 2 mM DTT, pH 6.8) supplemented with protease inhibitors (Complete; Roche 693124001). The homogenate was then incubated at 4°C for 20 min to depolymerize any remaining microtubules before being transferred to polycarbonate centrifuge bottles (16 × 76 mm; Beckman 355603) and centrifuged at 11,000 g (12,700 RPM) for 20 min at 4°C in an ultracentrifuge (Beckman XL100K) using a pre-chilled 70.1 rotor (Beckman 342184). The pellet was kept on ice. The supernatant was pooled in polycarbonate bottles and centrifuged again at 100,000 g (38,000 RPM) for 1 h at 4°C in a 70.1 Ti rotor. The pellets from the first and second centrifugations were resuspended in extraction buffer (10 ml per gram of tissue) (10 mM Tris-HCl, pH 7.4, 10% sucrose, 0.85 M NaCl, 1 tablet / 50 ml protease inhibitor (Roche, 4693124001), 1 mM EGTA, 1% phosphatase inhibitor (Sigma, P5726 and P0044)). The solution was then transferred to polycarbonate centrifuge bottles (16 x 76 mm; Beckman, 355603) and centrifuged at 15,000 g (14,800 RPM) for 20 min at 4°C in an ultracentrifuge (Beckman, XL100K) using a 70.1 Ti rotor. The pellets were stored at -80°C. 30% sarkosyl (Fluka analytical, 61747) was added to the supernatant to a final concentration of 1% and stirred at RT for 1 h. This solution was then centrifuged in a polycarbonate bottle at 100,000 g (38,000 RPM) in a 70.1 Ti rotor for 1 h at 4° C., and the pellet containing the PHF-enriched material was resuspended in 50 μl PBS / g of brain tissue.The resuspended PHFs were then sonicated on ice for 2 min at 30% amplitude with a 0.5 sec-on / 0.5 sec-off cycle. Aliquots were snap frozen and stored at -80°C.

[0709] 2.5.2.3 Compound Incubation and Readout α-synuclein aggregates derived from PD brains and tau PHF aggregates derived from AD brains were spotted on microarray slides. The slides spotted with α-synuclein aggregates were treated with 25 nM or 40 nM [ 3 H]-α-synuclein standards and a series of increasing concentrations of example compounds (non-radiolabeled) ranging from 100 nM and 0.05 nM to 2 μM. Additionally, slides spotted with α-synuclein aggregates or tau PHF samples were incubated with [ 3 H] Compound-1, [ 3 H] Compound-17 and [ 3 H]-α-synuclein standard or [ 3 The slides were incubated with [H]tau standard compounds at concentrations ranging from 0.6 to 200 nM. After incubation, the slides were washed and scanned using a real-time autoradiography system (BeaQuant, ai4R). Signal quantification was performed using the image analysis software Beamage (ai4R). Nonspecific signals were determined using an excess of nonradiolabeled compounds (2 μM), and specific binding was calculated by subtracting the nonspecific signal from the total signal. Competition was calculated as a percentage, with 0% defined as specific binding in the presence of vehicle and 100% defined as the value obtained in the presence of excess nonradiolabeled a-syn standard compounds. Ki values ​​were calculated using GraphPad Prism 7 by applying a nonlinear regression curve fit using a one-site specific binding model. All measurements were performed with at least two technical replicates. Kd and R 2 Values ​​were obtained by fitting the specific binding data to nonlinear regression analysis using a one-site specific binding model in GraphPad Prism.

[0710] Results: Example compounds were evaluated for selectivity for TDP-43 over Abeta and a-synuclein. To assess selectivity over Abeta, inhibitor constant (Ki) values ​​were calculated as [ 3 H]A beta standard compound ([ 3 H]A beta) in AD brain homogenates (Table 7). As can be seen from the high Ki values ​​in Table 7, the compounds of the present invention show good selectivity for TDP-43 over Abeta.

[0711] [Table 8]

[0712] To assess selectivity over a-synuclein, the potency of example compound 1 to compete with the binding of a [H]-standard a-syn compound in insoluble fractions derived from PD brain was measured, and compound 1 exhibited a K of >1000 nM in AD brain homogenates and a K of >500 nM in insoluble fractions derived from PD brain.

[0713] Furthermore, [ 3 H] Compound-1 and [ 3 The selectivity of [H] compound-17 over Abeta, a-synuclein, and tau was assessed by direct binding in AD brain homogenates (containing Abeta and tau, Figures 5a-b), a-synuclein aggregates from PD brains (Figures 5c-d), and tau PHF aggregates from AD brains (Figures 5e-f). In any of these preparations, [H] compound-17 was shown to be highly selective for TDP-43 over Abeta, a-synuclein, and tau. 3 H] Compound-1 also 3 H] Compound-17 also did not detect significant binding.

[0714] The results demonstrate that compounds according to the invention exhibit good TDP-43 selectivity over Abeta, a-synuclein and tau.

[0715] 2.5.3. Evaluation of Target Engagement of Examples 1 and 17 with Abeta and Tau Aggregates in AD Tissue by Classical Autoradiography and High-Resolution Micro-Autoradiography 2.5.3.1 Classical autoradiography All tissues were collected from donors, whose written informed consent for brain autopsy and use of their materials and clinical information for research purposes was obtained from or by the respective brain banks. All samples were anonymized and coded. Frozen brain tissue blocks in which Abeta and tau pathology were identified during autopsy were processed using a cryotome to generate 10-μm-thick sections, which were then mounted on glass slides. Sections were stored at -80°C until use.

[0716] Brain sections were fixed in 4% formaldehyde on ice for 15 minutes at 4°C and washed three times with 1x PBS for 8 minutes at room temperature. Sections were blocked for 30 minutes in assay buffer (50 mM Tris-HCl, 0.9% NaCl + 0.1% BSA). Cold test compound (Compound 1) was diluted to 4 μM in assay buffer and applied to half of the sections for 40 minutes at room temperature to determine nonspecific binding. Only the other half of the sections was incubated with assay buffer (total binding). After this incubation, an equal volume of the corresponding tritiated compound (i.e., [ 3 H]-Compound 1, [ 3 H]-Compound 17, [ 3 H]-A beta standard compound or [ 3 [H]-tau (standard compound) was diluted to 10 nM, added to the sections, and incubated for 2 h at RT, followed by washing as follows: once for 1 min in ice-cold 50 mM Tris-HCl buffer, pH 7.4, twice for 1 min in ice-cold PBS, once for 1 min in ice-cold 50 mM Tris-HCl buffer, pH 7.4, and finally a brief rinse in ice-cold distilled water.

[0717] The sections were then dried under airflow and mounted with copper-based adhesive strips. Bound radioactivity was quantified by exposing the slides for 2 hours in a real-time autoradiography system (Beaquant™, AI4R) with the measurement settings set for tritium. Images were acquired using Beavacq software (AI4R) and subsequently quantified using Beamage image analysis software (AI4R).

[0718] result: Example compounds 1 and 17 were evaluated for their binding on human tissue sections from AD cases containing abundant Abeta and tau pathology (Figures 6a and 6b, respectively). Images of total binding (first row) and nonspecific binding (middle row) are shown. 3 H]-Compound 1 or 17 (first vertical column), [ 3 H]-A beta standard compound (second column) or [ 3 H]-tau standard compound (last column). Immunofluorescence images (bottom row) of pTDP-43 (first column), Abeta (second column), or ptau (last column). Example compounds 1 and 17 showed no signal in tissue under any of the conditions tested (radiolabeled compound alone or self-competition), which correlates with the absence of pTDP-43 immunolabeling in adjacent sections. Both Abeta and tau standards showed overall binding signals that were displaced under self-competition, while correlating with ptau and Abeta immunolabeling in adjacent sections of tissue. The results further demonstrate that compounds according to the present invention exhibit good TDP-43 selectivity over Abeta and tau.

[0719] 2.5.3.2 High-resolution micro-autoradiography The protocol was adapted from Marquie et al., 2015. AD slices were treated with 20 nM [ 3 H]-Compound 1, [ 3 H]-compound 17 or [ 3The sections were incubated with [H]-tau standard compounds for 1 hour at room temperature (RT). The sections were then washed as follows: once for 1 minute in ice-cold 50 mM Tris-HCl buffer, pH 7.4, twice for 1 minute in ice-cold 70% ethanol, once for 1 minute in ice-cold 50 mM Tris-HCl buffer, pH 7.4, and finally briefly rinsed in ice-cold distilled water. The sections were then dried and then exposed to Ilford Nuclear Emulsion Type K5 (Agar Scientific, AGP9281) in a light-proof slide storage box. After 5 days, the sections were developed by successively immersing them in the following solutions: 1.) Ilford Phenisol Developer (diluted 1:5 in H2O, Agar Scientific, AGP9106), 2.) Ilfostop solution (diluted 1:20 in H2O, Agar Scientific, AGP9104), 3.) Ilford Hypam Fixer (diluted 1:5 in H2O, Agar Scientific, AGP9183), and finally rinsed in H2O.

[0720] Thioflavin S staining was performed on adjacent sections. For image acquisition, sections were mounted using ProLong Gold Antifade reagent (Invitrogen P36930) and imaged on a Panoramic 150 slide scanner (3DHistech) with a 20x objective, which captures bright-field and fluorescent images separately.

[0721] result: Incubated on human brain slices [ 3 H]-Compound 1, [ 3 H]-compound 17 and [ 3 The micro-autoradiography signal from the [H]-tau standard was assessed in the form of accumulated silver granules in areas rich in tau tangles. The results are shown in Figure 6c ([ 3 H]-Compound 1) and Figure 6d ([ 3 Left: Thioflavin S staining in the same tissue labeled tau aggregates. Right: [H]-compound 17) in the same tissue. 3 H]-Compound 1 or [3 H]-compound 17 and [ 3 Image of silver granule deposition in [H]-tau standard compound. 3 H]-Compound 1 or [ 3 H]-Compound 17 was used to 3 No silver granules accumulated in the tau tangles compared to the [H]-tau standard. The bottom row images are zoomed-in images from the area indicated by the square in the top row image. * NFT: neurofibrillary tangles. Scale bars are 200 μm (upper row) and 50 μm (lower row, FIG. 6c) or 100 μm (lower row, FIG. 6d). As shown in FIG. 6c and FIG. 6d, the [ 3 H]-Compound 1 or [ 3 Incubation with [H]-compound 17 in adjacent sections yielded [ 3 H]-tau reference compounds showed target engagement in regions with tau tangles as determined by Thioflavin S staining, but showed no accumulation of silver granules. The results further demonstrate that compounds according to the present invention exhibit good TDP-43 selectivity over tau.

[0722] 2.6 PK studies in healthy monkeys In non-human primates (NHPs) [ 18 F]-labeled compound 1([ 18 F] Compound 1) (4.2 mCi) or [ 18 F]-labeled compound 17([ 18 [F]-compound 17) (5.3 mCi) was injected intravenously (iv). PET scans of the monkeys were performed using a Siemens Focus 220. PET acquisition started just before the injection of the radiation dose. Images were generated as a 120-minute dynamic scan focused on the head. 18 F]-Compound 1 and [ 18 [F]-Compound 17 rapidly entered the brain (5.5 and 4.5 min after injection, respectively) and showed robust uptake in the whole brain with SUVmax of 2.7 and 1.1, respectively (Figures 7a-b). 18 F]-Compound 1 and [ 18F]-Compound 17 showed rapid washout from peak to half-peak of <17 min and <13 min, respectively. These data demonstrate that the [F]-Compound 17 of the present invention is suitable for use as a brain PET agent in humans. 18 F]-Compound 1 and [ 18 [F]-Compound 17 in non-human primates.

Claims

1. Formula (I) 【Chemical 1】 (In the formula, n is 1 or 2, R 1 is H, hydroxy (C 1 ~C 4 ) alkyl or F; R 2 is optionally F, NH 2 , CN and / or CH 3 wherein the heterocyclic ring contains one or more heteroatoms selected from N, O and S), or a detectably labeled compound, stereoisomer, polymorph, racemic mixture, tautomer, pharmaceutically acceptable salt, prodrug, hydrate or solvate thereof.

2. R 1 The compound having formula (I) according to claim 1, wherein is H or F.

3. R 2 but 【Chemistry 2】 (In the formula, R 3 is F and R 4 NH 2 and R 7 is H and R 8 is H, R 3 NH 2 and R 4 is F and R 7 is H and R 8 is H, R 3 is CN and R 4 NH 2 and R 7 is H and R 8 is H, R 3 is H and R 4 NH 2 and R 7 is H and R 8 is CN, R 3 is H and R 4 NH 2 and R 7 is H and R 8 is F, or R 3 is H and R 4 NH 2 and R 7 is H and R 8 is CN), and or R 2 but, 【Chemistry 3】 (wherein X is N and R 5 is CH 3 or H), or R 2 but, 【Chemistry 4】 (In the formula, R 9 are H, F, and CH 3 and NH 2 and preferably H; or R 2 but, 【Chemistry 5】 and or R 2 but, 【Chemistry 6】 and or R 2 but, 【Chemistry 7】 and or R 2 but, 【Chemistry 8】 and or R 2 but, 【Chemistry 9】 and or R 2 but, 【Chemistry 10】 and or R 2 but, 【Chemistry 11】 and or R 2 but, 【Chemistry 12】 3. The compound according to claim 1 or 2, wherein

4. R 2 but, 【Chemistry 13】 (In the formula, R 3 is F and R 4 NH 2 and R 7 is H and R 8 is H), 【Chemistry 14】 (wherein X is N and R 5 is CH 3 is) 【Chemistry 15】 (In the formula, R 9 is selected from H and F) 【Chemistry 16】 4. The compound according to any one of claims 1 to 3, wherein

5. 【Chemical 17】 4. The compound according to any one of claims 1 to 3, selected from:

6. 6. The compound of any one of claims 1 to 5, comprising a detectable label.

7. Detectable label 3 H or 18 7. The compound of claim 6, wherein:

8. Expression (IT) 【Chemistry 18】 (Wherein n is 1 or 2, R 1 But H, hydroxy (C 1 ~C 4 ) alkyl or F; T is 3 H, R 2 but 【Chemistry 19】 (In the formula, R 3 is F and R 4 Ga-NH 2 and R 7 and R 8 at least one of which is T and, if applicable, the other is H, preferably R 7 is T and R 8 is T, R 6 is T), or R 2 but 【Chemistry 20】 (In the formula, R 4 Ga-NH 2 and R 8 is CN and R 3 or R 7 at least one of which is T and, if applicable, the other is H, preferably R 7 is T and R 3 is T, or R 7 is T and R 3 is H, R 6 is T), Or, R 2 but 【Chemical 21】 (wherein X is N and R 5 CT 3 and R 6 is H), or R 2 but 【Chemical 22】 (In the formula, R 12 is T), or R 2 but 【Chemical 23】 (In the formula, R 12 is T), or R 2 but 【Chemistry 24】 (In the formula, R 12 is T), or R 2 but 【Chemistry 25】 (In the formula, R 12 is T) is 8. The compound of claim 7, or a stereoisomer, polymorph, racemic mixture, tautomer, pharmaceutically acceptable salt, prodrug, hydrate or solvate thereof, having the formula:

9. Formula (I-T') 【Chemical Formula 26】 (In the formula, n is 1 or 2, R 1 is substituted by one or two, preferably two, T's; 1 ~C 4 ) alkyl, T 3 H, R 2 , but optionally, F, NH 2 , CN and / or CH 3 and wherein the heterocyclic ring contains one or more heteroatoms selected from N, O and S.

8. The compound of claim 7, or a stereoisomer, polymorph, racemic mixture, tautomer, pharmaceutically acceptable salt, prodrug, hydrate or solvate thereof, having the formula:

10. The compound is 【Chemical 27】 (Where T is 3 H) 9. The compound of claim 8, selected from:

11. The compound is 【Chemical formula 28】 (Where T is 3 H) 10. The compound of claim 9, selected from:

12. Formula (IF) 【Chemical 29】 (In the formula, n is 1 or 2, R 1 but 18 F, R 2 but 【Chemistry 30】 (wherein X is N and R 5 is CH 3 is) or 【Chemical 31】 (In the formula, R 9 is H) is 8. The compound of claim 7, or a stereoisomer, polymorph, racemic mixture, tautomer, pharmaceutically acceptable salt, prodrug, hydrate or solvate thereof, having the formula:

13. Formula (IF) 【Chemical 32】 (In the formula, n is 1 or 2, R 1 is H or hydroxy (C 1 ~C 4 ) alkyl, R 2 but, 【Chemical 33】 (In the formula, R 9 is H) is 8. The compound of claim 7, or a stereoisomer, polymorph, racemic mixture, tautomer, pharmaceutically acceptable salt, prodrug, hydrate or solvate thereof, having the formula:

14.

34. 13. The compound of claim 12, wherein:

15. 【Catalog 35】 14. The compound of claim 13, wherein:

16. 16. A diagnostic composition comprising a compound according to any one of claims 1 to 15, and optionally at least one physiologically acceptable carrier, diluent, adjuvant and / or excipient.

17. A compound according to any one of claims 1 to 15 or a diagnostic composition according to claim 16 for use in a diagnostic method.

18. 17. The compound of any one of claims 1 to 15, or the diagnostic composition of claim 16, wherein the compound or diagnostic composition is for use in imaging TDP-43 aggregates, in particular wherein the imaging is performed by positron emission tomography.

19. 17. The compound of any one of claims 1 to 15, or the diagnostic composition of claim 16, wherein the compound or diagnostic composition is for use in diagnosing a disease, disorder or abnormality associated with TDP-43 aggregates, in particular wherein the diagnosis is carried out by positron emission tomography.

20. 1. A method for diagnosing a disease, disorder, or abnormality associated with TDP-43 aggregates, or a predisposition thereto, in a subject, comprising: (a) administering to a subject a compound according to any one of claims 1 to 15 or a diagnostic composition according to claim 16 comprising a compound according to any one of claims 1 to 15; (b) binding the compound to TDP-43 aggregates; and (c) detecting a compound bound to TDP-43 aggregates A method comprising:

21. 21. A method for diagnosing a disease, disorder, or abnormality associated with TDP-43 aggregates, or a predisposition thereto, according to claim 20, comprising: (d) generating an image showing the location and / or amount of the compound bound to the TDP-43 aggregates; (e) optionally, comparing the generated image with a control image of a healthy control subject, wherein an increase in binding signal indicates that the subject is suffering from or at risk of developing a disease, disorder, or condition associated with TDP-43 aggregates. The method further comprises:

22. 1. A method of positron emission tomography (PET) imaging of TDP-43 aggregates in tissue of a subject, comprising: (a) administering to a subject a compound according to any one of claims 1 to 15 or a diagnostic composition according to claim 16 comprising a compound according to any one of claims 1 to 15; (b) binding the compound to TDP-43 aggregates; and (c) detecting the compound bound to the TDP-43 aggregates by collecting positron emission tomography (PET) images of the subject's tissue. A method comprising:

23. 23. The method of positron emission tomography (PET) imaging of TDP-43 aggregates in a tissue of a subject according to claim 22, wherein the tissue is a tissue of the central nervous system (CNS), an eye tissue or a brain tissue, preferably the tissue is a brain tissue.

24. 1. A method for detecting, and optionally quantitating, TDP-43 aggregates in tissue of a subject, comprising: (a) contacting a sample suspected of containing TDP-43 aggregates, or a specific body part or body region, with a compound according to any one of claims 1 to 15, or with the diagnostic composition according to claim 16 comprising a compound according to any one of claims 1 to 15; (b) binding the compound to TDP-43 aggregates; (c) detecting the compound binding to the TDP-43 aggregates using positron emission tomography; and (d) optionally, quantifying the amount of compound bound to the TDP-43 aggregates. A method comprising:

25. 1. A method of collecting data for diagnosing a disease, disorder, or condition associated with, or a predisposition to, TDP-43 aggregates, comprising: (a) contacting a sample suspected of containing TDP-43 aggregates, or a specific body part or body region, with a compound according to any one of claims 1 to 15, or with the diagnostic composition according to claim 16 comprising a compound according to any one of claims 1 to 15; (b) binding the compound to TDP-43 aggregates; (c) detecting compounds that bind to TDP-43 aggregates; and (d) optionally correlating the presence or absence of the compound binding to the TDP-43 aggregates with the presence or absence of TDP-43 aggregates in the sample or in a particular body part or body region. A method comprising:

26. 1. A method of collecting data for monitoring the progression of a disease, disorder, or condition associated with TDP-43 aggregates in a patient, comprising: (a) contacting a sample, a specific body part or a body area suspected of containing TDP-43 aggregates with a compound according to any one of claims 1 to 15 or with the diagnostic composition according to claim 16 comprising a compound according to any one of claims 1 to 15; (b) binding the compound to TDP-43 aggregates; (c) detecting a compound that binds to the TDP-43 aggregates; (d) optionally correlating the presence or absence of the compound binding to TDP-43 aggregates with the presence or absence of TDP-43 aggregates in the sample or in a particular body part or region; and (e) optionally repeating steps (a) through (c), and, if present, optional step (d), at least once. A method comprising:

27. 1. A method for collecting data to predict the responsiveness of a patient suffering from a disease, disorder, or condition associated with TDP-43 aggregates to treatment with a pharmaceutical agent, comprising: (a) contacting a sample, a specific body part or a body area suspected of containing TDP-43 aggregates with a compound according to any one of claims 1 to 15 or with the diagnostic composition according to claim 16 comprising a compound according to any one of claims 1 to 15; (b) binding the compound to TDP-43 aggregates; (c) detecting a compound that binds to the TDP-43 aggregates; (d) optionally correlating the presence or absence of the compound binding to TDP-43 aggregates with the presence or absence of TDP-43 aggregates in the sample or in a particular body part or region; and (e) optionally repeating steps (a) through (c), and, if present, optional step (d), at least once. A method comprising:

28. Optionally, correlating the presence or absence of the compound binding to TDP-43 aggregates with the presence or absence of TDP-43 aggregates in the sample or in a particular body part or body region comprises: - determining the amount of the compound bound to the TDP-43 aggregates; - correlating the amount of compound bound to TDP-43 aggregates with the amount of TDP-43 aggregates in the sample or in a particular body part or body region; - optionally comparing the amount of compound bound to TDP-43 aggregates in the sample or in a particular body part or body region with a normal control value in a healthy control subject.

28. The method of any one of claims 25 to 27, comprising:

29. Use of a compound according to any one of claims 1 to 15 as a biomarker for TDP-43 aggregates or a biomarker for TDP-43 proteinopathy.

30. 16. Use of a compound according to any one of claims 1 to 15 as a diagnostic agent or tool for TDP-43 proteinopathy.

31. 16. A compound according to any one of claims 1 to 15 for use as an in vitro analytical standard or an in vitro screening tool.

32. The disease, disorder, or abnormality associated with TDP-43 aggregates or TDP-43 proteinopathy may be frontotemporal dementia (FTD, e.g., sporadic or familial, with or without motor neuron disease (MND), with progranulin (GRN) mutations, with C9orf72 mutations, with TARDBP mutations, with valosin-containing protein (VCP) mutations, linked to chromosome 9p, corticobasal degeneration, frontotemporal lobar degeneration (FTLD), including frontotemporal lobe dementia with TDP-43 or with ubiquitin-positive TDP-43 inclusions. Frontotemporal lobar degeneration (FTLD-TDP), argyrophilic grain dementia, Pick's disease, semantic primary progressive aphasia (svPPA), behavioral variant of FTD (bvFTD), non-fluent primary progressive aphasia (e.g., nfvPPA), amyotrophic lateral sclerosis (ALS, e.g., sporadic ALS with TARDBP mutations and sporadic ALS with angiogenin (ANG) mutations), Alexander disease (AxD), limbic-predominant age-related TDP-43 encephalopathy (LATE), chronic traumatic encephalopathy, Perry syndrome, Alzheimer's disease (AD, including sporadic and familial forms of AD), Down's syndrome, familial British type Dementia, polyglutamine diseases (Huntington's disease and spinocerebellar ataxia type 3 (SCA3; also known as Machado-Joseph disease)), hippocampal sclerosis dementia and myopathy (sporadic inclusion body myositis, inclusion body myopathy with mutations in valosin-containing protein (VCP); also Paget's disease of bone and frontotemporal dementia), oculopharyngeal muscular dystrophy with rimmed vacuoles, myofibrillar myopathy with mutations in the myotilin (MYOT) gene or in the gene encoding desmin (DES), traumatic brain injury (TBI), dementia with Lewy bodies ( 29. The compound for use or the diagnostic composition for use according to claim 18 or 19, or the method according to any one of claims 20 to 28, wherein the disease, disorder or abnormality associated with TDP-43 aggregates, or the TDP-43 proteinopathy is selected from frontotemporal dementia (FTD), amyotrophic lateral sclerosis (ALS), Alzheimer's disease (AD), Parkinson's disease (PD), chronic traumatic encephalopathy (CTE), and limbic-predominant age-related TDP-43 encephalopathy (LATE).

33. 33. The compound for use, or diagnostic composition for use, or method according to claim 32, wherein the disease, disorder or abnormality associated with TDP-43 aggregates, or TDP-43 proteinopathy, is amyotrophic lateral sclerosis (ALS).

34. 33. The compound for use, or diagnostic composition for use, or method according to claim 32, wherein the disease, disorder or abnormality associated with TDP-43 aggregates, or TDP-43 proteinopathy, is Alzheimer's disease (AD).

35. 33. The compound for use, or diagnostic composition for use, or method according to claim 32, wherein the disease, disorder, or abnormality associated with TDP-43 aggregates, or TDP-43 proteinopathy, is frontotemporal dementia (FTD), including frontotemporal lobar dementia associated with TDP-43, or frontotemporal lobar degeneration with TDP-43 inclusions (FTLD-TDP).

36. 33. The compound for use, or diagnostic composition for use, or method according to claim 32, wherein the disease, disorder or abnormality associated with TDP-43 aggregates, or TDP-43 proteinopathy, is limbic-predominant age-related TDP-43 encephalopathy (LATE).

37. 33. The compound for use, or diagnostic composition for use, or method according to claim 32, wherein the disease, disorder, or abnormality associated with TDP-43 aggregates, or TDP-43 proteinopathy, is frontotemporal dementia (FTD) with a progranulin (GRN) mutation, or frontotemporal dementia (FTD) with a C9orf72 mutation.

38. Formula (II) 【Chemical 36】 (In the formula, n is 1 or 2, R 1 is H, hydroxy (C 1 ~C 4 ) alkyl or F) or a stereoisomer, polymorph, racemic mixture, tautomer, pharmaceutically acceptable salt, prodrug, hydrate or solvate thereof, or a mixture thereof.

39. Formula (III) 【Chemical 37】 (In the formula, Z is selected from C—Br, CI and CH; n is 1 or 2, R 1 is H, hydroxy (C 1 ~C 4 ) alkyl or F; R 11 is optionally Br, I, F, NH 2 , CN and / or CH 3 wherein the heterocyclic ring contains one or more heteroatoms selected from N, O and S; Z and / or R 11 at least one of which contains Br or I or a stereoisomer, polymorph, racemic mixture, tautomer, pharmaceutically acceptable salt, prodrug, hydrate or solvate thereof, or a mixture thereof.

40. Formula (IV) 【Chemical 38】 (In the formula, n is 1 or 2, R 2 teeth, 【Chemical Formula 39】 (In the formula, R 9 is H), R 14 is the leaving group (LG) A compound having the formula:

41. Formula (V) 【Chemistry 40】 (In the formula, n is 1 or 2, R 1 is H or hydroxy (C 1 ~C 4 ) alkyl, R 2 teeth, 【Chemistry 41】 (In the formula, R 9 is H), R 14 is the leaving group (LG) A compound having the formula:

42. Formula (III) 【Chemistry 42】 (In the formula, n is 1 or 2, R 15 -COOR' or -C 1 ~C 3 alkylCOOR', wherein R' is selected from C 1 ~C 3 is alkyl, R 2 is optionally F, NH 2 , CN and / or CH 3 and wherein the heterocyclic ring contains one or more heteroatoms selected from N, O and S. or a stereoisomer, polymorph, racemic mixture, tautomer, pharmaceutically acceptable salt, prodrug, hydrate or solvate thereof.

43. 16. A kit for preparing a radiopharmaceutical preparation comprising a detectably labelled precursor of a compound according to any one of claims 6 to 15, wherein the precursor is a compound of formula (II) according to claim 38, a compound of formula (III) according to claim 39 or claim 42, a compound of formula (IV) according to claim 40, or a compound of formula (V) according to claim 41.

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